BioMed Research International: Parasitology The latest articles from Hindawi Publishing Corporation © 2016 , Hindawi Publishing Corporation . All rights reserved. Detection of Natural Toxoplasma gondii Infection in Chicken in Thika Region of Kenya Using Nested Polymerase Chain Reaction Thu, 17 Nov 2016 12:59:10 +0000 The detection of Toxoplasma gondii in free-range chickens is a good indicator of possible risk to human beings. The aim of this study was to investigate the occurrence of T. gondii in free-range chicken using polymerase chain reaction (PCR). Brain samples from 105 free-range chickens from three administrative areas in Thika region, Kenya, were collected, DNA-extracted, and analyzed using PCR to detect presence of T. gondii. The overall prevalence of T. gondii in all the three areas was 79.0% (95% CI: 70.0–86.4%) and the prevalence across the three areas was not significantly different (; ). Female chickens had higher (79.4%) prevalence than males (78.6%), although the difference was not significant (, = 0.01). However, chickens that were more than 2 years old had significantly (; = 11.87) higher prevalence compared to younger ones. The study indicates that there was a high occurrence of T. gondii infection in free-range chickens from Thika region and that the infection rate is age dependent. Further studies should be carried out to determine the possible role of roaming chickens in the epidemiology of the disease among humans in the area. John Mokua Mose, John Maina Kagira, Simon Muturi Karanja, Maina Ngotho, David Muchina Kamau, Adele Nyambura Njuguna, and Naomi Wangari Maina Copyright © 2016 John Mokua Mose et al. All rights reserved. Characterization of South American Snails of the Genus Biomphalaria (Basommatophora: Planorbidae) and Schistosoma mansoni (Platyhelminthes: Trematoda) in Molluscs by PCR-RFLP Thu, 17 Nov 2016 11:20:29 +0000 The identification of snails of the genus Biomphalaria can be done using morphological characteristics which depends on the size of the snails and skill and knowledge of researcher. These methods sometimes are not adequate for identification of species. The PCR-RFLP, using the ITS region of the rDNA, has been used to identify Brazilian species of the genus Biomphalaria. Nevertheless, there is a lack of information about snails from other Latin American countries. In addition, some snails may be infected by Schistosoma mansoni and when submitted to PCR-RFLP they show molecular profiles different from those previously standardized for the other mollusc species. In this work the molecular profiles of 15 species and the subspecies were established by PCR-RFLP of ITS-rDNA with the enzyme DdeI. Moreover, the molecular profiles of host species, B. glabrata, B. straminea, B. tenagophila, and B. prona, infected by S. mansoni were also established. The molluscs were dissected to permit morphological identification. These results contribute to a correct identification of snails of the genus Biomphalaria and detection of these snails infected by S. mansoni. Roberta Lima Caldeira, Tatiana Maria Teodoro, Liana Konovaloff Jannotti-Passos, Pollanah M. Lira-Moreira, Christiane De Oliveira Goveia, and Omar dos Santos Carvalho Copyright © 2016 Roberta Lima Caldeira et al. All rights reserved. Evaluation of Lethal Giant Larvae as a Schistosomiasis Vaccine Candidate Wed, 09 Nov 2016 09:19:30 +0000 Schistosomiasis is a neglected tropical disease of humans, and it is considered to be the second most devastating parasitic disease after malaria. Eggs produced by normally developed female worms are important in the transmission of the parasite, and they responsible for the pathogenesis of schistosomiasis. The tumor suppressor gene lethal giant larvae (lgl) has an essential function in establishing apical-basal cell polarity, cell proliferation, differentiation, and tissue organization. In our earlier study, downregulation of the lgl gene induced a significant reduction in the egg hatching rate of Schistosoma japonicum (Sj) eggs. In this study, the Sjlgl gene was used as a vaccine candidate against schistosomiasis, and vaccination achieved and maintained a stable reduction of the egg hatching rate, which is consistent with previous studies, in addition to reducing the worm burden and liver egg burden in some trials. Yufan Cao, Hongbin Qiao, Yanli Shi, Yu Han, Jinming Liu, Hao Li, Ke Lu, Jiaojiao Lin, and Yamei Jin Copyright © 2016 Yufan Cao et al. All rights reserved. Climate Change Is Increasing the Risk of the Reemergence of Malaria in Romania Wed, 26 Oct 2016 14:22:32 +0000 The climatic modifications lead to global warming; favouring the risk of the appearance and development of diseases are considered until now tropical diseases. Another important factor is the workers’ immigration, the economic crisis favouring the passive transmission of new species of culicidae from different areas. Malaria is the disease with the widest distribution in the globe. Millions of people are infected every year in Africa, India, South-East Asia, Middle East, and Central and South America, with more than 41% of the global population under the risk of infestation with malaria. The increase of the number of local cases reported in 2007–2011 indicates that the conditions can favour the high local transmission in the affected areas. In the situation presented, the establishment of the level of risk concerning the reemergence of malaria in Romania becomes a priority. Larisa Ivanescu, Ilie Bodale, Simin-Aysel Florescu, Constantin Roman, Dumitru Acatrinei, and Liviu Miron Copyright © 2016 Larisa Ivanescu et al. All rights reserved. Triflumuron Effects on the Physiology and Reproduction of Rhodnius prolixus Adult Females Sun, 16 Oct 2016 14:18:28 +0000 We evaluated the efficacy of the growth regulator triflumuron (TFM) in inducing mortality and disrupting both oviposition and egg hatching in Rhodnius prolixus adult females. TFM was administered via feeding, topically or by continuous contact with impregnated surfaces. Feeding resulted in mild biological effects compared with topical and impregnated surfaces. One day after treatment, the highest mortality levels were observed with topical surface and 30 days later both topical and impregnated surfaces induced higher mortalities than feeding. Oral treatment inhibited oviposition even at lower doses, and hatching of eggs deposited by treated females was similarly affected by the three delivery modes. Topical treatment of eggs deposited by nontreated females significantly reduced hatching. However, treatment per contact of eggs oviposited by untreated females did not disrupt eclosion. Additionally, oral treatment increased the number of immature oocytes per female, and topical treatment reduced the mean size of oocytes. TFM also affected carcass chitin content, diuresis, and innate immunity of treated insects. These results suggest that TFM acts as a potent growth inhibitor of R. prolixus adult females and has the potential to be used in integrated vector control programs against hematophagous triatomine species. Bianca Santos Henriques, Fernando Ariel Genta, Cícero Brasileiro Mello, Lucas Rangel Silva, Thaís Franco Codogno, Alyne F. R. Oliveira, Lourena Pinheiro Marinho, Denise Valle, José Bento Pereira Lima, Denise Feder, Marcelo Salabert Gonzalez, and Patricia Azambuja Copyright © 2016 Bianca Santos Henriques et al. All rights reserved. Evaluation of Protective Immune Responses Induced by Recombinant TrxLp and ENO2 Proteins against Toxoplasma gondii Infection in BALB/c Mice Mon, 10 Oct 2016 14:09:59 +0000 Toxoplasma gondii is an obligate intracellular parasitic protozoan that can infect almost all species of warm-blooded animals. As any chemical-based drugs could not act against the tissue cyst stage of T. gondii, vaccination may be one of the ideal control strategies. In the present study, two new vaccine candidates, named TgENO2 and TgTrxLp, were purified from Escherichia coli with pET-30a(+) expression system and then were injected into BALB/c mice to evaluate the protective efficacy against acute and chronic toxoplasmosis. The results showed that both the recombinant proteins, either alone or in combination, could elicit strong humoral and cellular immune responses with a higher level of IgG antibodies, IFN-γ, IL-2, CD4+, and CD8+ T cells as compared to those in mice from control groups. After acute challenge with tachyzoites of the GJS strain, mice immunized with rTgTrxLp ( d), rTgENO2 ( d), and rTgTrxLp + rTgENO2 ( d) proteins showed significantly longer survival time than those that received Freund’s adjuvant ( d) and PBS ( d) ( = 9.687, df = 4, ). The protective immunity of rTgTrxLp, rTgENO2, and rTgTrxLp + rTgENO2 proteins against chronic T. gondii infection showed 69.77%, 58.14%, and 20.93% brain cyst reduction as compared to mice that received PBS. The present study suggested that both TgENO2 and TgTrxLp were potential candidates for the development of multicomponent vaccines against toxoplasmosis. Meng Wang, Xiao-Yu Yang, Nian-Zhang Zhang, De-Lin Zhang, and Xing-Quan Zhu Copyright © 2016 Meng Wang et al. All rights reserved. Efficacy and Safety of a Permethrin-Fipronil Spot-On Solution (Effitix®) in Dogs Naturally Infested by Ticks in Europe Thu, 15 Sep 2016 08:04:34 +0000 Effitix is a new broad spectrum product based on the combination of fipronil 6.1% and permethrin 54.5% in a solution for spot-on application. It has been shown to be safe and efficacious in dogs in controlling tick, flea, sandfly, and mosquito infestations in laboratory conditions. The aim of this controlled, randomised study was to assess its safety and efficacy against natural tick infestations in field conditions. One hundred eighty-two privately owned dogs were included in France and Germany: 123 dogs were treated on day 0 with the permethrin-fipronil combination (Effitix) and 59 with a permethrin-imidacloprid combination (Advantix®). Tick counts were conducted on days 0 (before treatment), 7, 14, 21, and 28. The percentages of efficacy on days 7, 14, 21, and 28 were, respectively, 91.2%, 97%, 98.3%, and 96.7% with Effitix and were 94.8%, 96.9%, 95.7%, and 94.6% with Advantix. Very few adverse events were reported. Most were not serious and/or not related to the treatment with pruritus being the most common. One administration of Effitix was highly effective and safe to treat and control tick infestations for four weeks in field conditions and had a similar efficacy as the permethrin-imidacloprid combination for all visits. Christelle Navarro, Nadège Reymond, Nolwenn Crastes, and Stéphane Bonneau Copyright © 2016 Christelle Navarro et al. All rights reserved. Immunopathology of Parasitic Infections and Therapeutic Approaches in Humans and Animals Wed, 31 Aug 2016 11:22:07 +0000 Orlando Paciello, Chiara Palmieri, Iwona Otrocka-Domagala, Laura Rinaldi, Jorge Morales-Montor, and Peter Geldhof Copyright © 2016 Orlando Paciello et al. All rights reserved. Acute Oral Mammalian Toxicity and Effect of Solvents on Efficacy of Maerua edulis (Gilg. & Ben.) De Wolf against Rhipicephalus (Boophilus) decoloratus Koch, 1844 (Acarina: Ixodidae), Tick Larvae Wed, 31 Aug 2016 06:23:47 +0000 Efficacy and toxicity of aqueous and organic solvents extracts of Maerua edulis against ticks and mice, respectively, were determined. Ground leaves were extracted separately using cold water, cold water plus surfactant (1% v/v liquid soap), hot water plus surfactant, hexane, or methanol to make 25% w/v stock solutions from which serial dilutions of 5, 10, 20, and 25% were made. For each concentration, 20 Rhipicephalus decoloratus tick larvae were put in filter papers impregnated with extracts and incubated for 48 h at 27°C and 85–90% RH for mortality observation after 24 h and 48 h. In the toxicity experiment, hot water plus surfactant treatments of 5, 10, 20, and 25% (w/v) M. edulis were administered in suspension per os to sexually mature Balb/C mice and observed for clinical signs and mortality for 72 h. Larvae mortality was highest (>98%) in methanol-extracted M. edulis treatments (20 and 25%), which was not different from the amitraz-based control (Tickbuster®). Mortality was also higher in the hot water than in cold water plus surfactant treatments (). No postadministration adverse health effects were observed in the mice. These results suggest that M. edulis is an effective tick remedy best extracted using methanol or hot water plus surfactant. Emmanuel T. Nyahangare, Brighton M. Mvumi, and Tatenda Maramba Copyright © 2016 Emmanuel T. Nyahangare et al. All rights reserved. Role of Macrophages in the Repair Process during the Tissue Migrating and Resident Helminth Infections Thu, 25 Aug 2016 17:06:50 +0000 The Th1/Th2/Th17 balance is a fundamental feature in the regulation of the inflammatory microenvironment during helminth infections, and an imbalance in this paradigm greatly contributes to inflammatory disorders. In some cases of helminthiasis, an initial Th1 response could occur during the early phases of infection (acute), followed by a Th2 response that prevails in chronic infections. During the late phase of infection, alternatively activated macrophages (AAMs) are important to counteract the inflammation caused by the Th1/Th17 response and larval migration, limiting damage and repairing the tissue affected. Macrophages are the archetype of phagocytic cells, with the primary role of pathogen destruction and antigen presentation. Nevertheless, other subtypes of macrophages have been described with important roles in tissue repair and immune regulation. These types of macrophages challenge the classical view of macrophages activated by an inflammatory response. The role of these subtypes of macrophages during helminthiasis is a controversial topic in immunoparasitology. Here, we analyze some of the studies regarding the role of AAMs in tissue repair during the tissue migration of helminths. Berenice Faz-López, Jorge Morales-Montor, and Luis I. Terrazas Copyright © 2016 Berenice Faz-López et al. All rights reserved. Comparative Assessment of Health Benefits of Praziquantel Treatment of Urogenital Schistosomiasis in Preschool and Primary School-Aged Children Thu, 18 Aug 2016 12:20:31 +0000 Schistosomiasis is a major public health problem in Africa. However, it is only recently that its burden has become recognised as a significant component impacting on the health and development of preschool-aged children. A longitudinal study was conducted in Zimbabwean children to determine the effect of single praziquantel treatment on Schistosoma haematobium-related morbidity markers: microhaematuria, proteinuria, and albuminuria. Changes in these indicators were compared in 1–5 years versus 6–10 years age groups to determine if treatment outcomes differed by age. Praziquantel was efficacious at reducing infection 12 weeks after treatment: cure rate = 94.6% (95% CI: 87.9–97.7%). Infection rates remained lower at 12 months after treatment compared to baseline in both age groups. Among treated children, the odds of morbidity at 12 weeks were significantly lower compared to baseline for proteinuria: odds ratio (OR) = 0.54 (95% CI: 0.31–0.95) and albuminuria: OR = 0.05 (95% CI: 0.02–0.14). Microhaematuria significantly reduced 12 months after treatment, and the effect of treatment did not differ by age group: OR = 0.97 (95% CI: 0.50–1.87). In conclusion, praziquantel treatment has health benefits in preschool-aged children exposed to S. haematobium and its efficacy on infection and morbidity is not age-dependent. Welcome M. Wami, Norman Nausch, Nicholas Midzi, Reggis Gwisai, Takafira Mduluza, Mark E. J. Woolhouse, and Francisca Mutapi Copyright © 2016 Welcome M. Wami et al. All rights reserved. Relationship between Antibody Levels, IgG Binding to Plasmodium falciparum-Infected Erythrocytes, and Disease Outcome in Hospitalized Urban Malaria Patients from Dakar, Sénégal Wed, 03 Aug 2016 11:06:35 +0000 Background. Management of clinical malaria requires the development of reliable diagnostic methods and efficient biomarkers for follow-up of patients. Protection is partly based on IgG responses to parasite antigens exposed at the surface of infected erythrocytes (iRBCs). These IgG responses appeared low during clinical infection, particularly in severe disease. Methods. We analyzed the IgG binding capacity to the surface of live erythrocytes infected by knob positive FCR3 strain. Sera from 69 cerebral malaria (CM) and 72 mild malaria (MM) cases were analyzed by ELISA for IgG responses to five antigens from iRBC and by flow cytometry for IgG binding as expressed in labeling index ratio (LIR). The relationship between IgG levels, LIR, parasitemia, age, and the clinical outcomes was evaluated. Results. We found a significant decrease of LIR in adult CM fatal cases compared to surviving patients (). In MM, LIRs were correlated to IgG anti-iRBC and anti-PfEMP3/5 levels. In CM, no correlation was found between LIR, IgG levels, and parasitemia. Conclusion. The IgG binding assay was able to discriminate outcome of cerebral malaria cases and it deserves further development as a potential functional-associated assay for symptomatic malaria analysis. Babacar Mbengue, Mouhamadou Mansour Fall, Maguette Sylla Niang, Birahim Niang, Marie Louise Varela, Antoine Marie Diatta, Moustapha Mbow, Kantome Ndiaye, Rokhaya Ndiaye Diallo, Alioune Dieye, and Ronald Perraut Copyright © 2016 Babacar Mbengue et al. All rights reserved. Comparative Diagnosis of Serum IgG1 and Coproantigen ELISA for Fasciolosis Detection of Goats in Mexico Mon, 01 Aug 2016 08:06:55 +0000 The objective of present study was to determine the prevalence of natural caprine fasciolosis in the Mixteca region of Mexico using coproantigen and serum IgG1 ELISA tests for comparative purposes. A total of 1070 serum and faecal samples were analyzed for IgG1 antibodies and coproantigens, using ELISA with E/S products as antigen and a monoclonal antibody-based sandwich ELISA. Prevalence of 73.46% was found using the serological ELISA and a percentage of 77.20 was found for coproantigen ELISA. The diagnostic sensitivity and specificity for serum ELISA were 86.7% and 96.4%, and for the coproantigen ELISA they were 93.1% and 97.8%, respectively. The seropositive samples were further categorized as low, medium, or high positivity. Results show a great proportion of low and medium positive goats when the serum ELISA test was used. Correlation coefficients between coproantigens and seropositivity were statistically significant () for low seropositivity () and medium seropositivity (). The accuracy of faecal antigen ELISA was higher compared to indirect ELISA serological test. Two ELISAs were shown to be useful for demonstrating the current status of F. hepatica infection in the endemic areas and can be employed in studies on epidemiology as well as anthelmintics treatment for preventing economic loss and the risk of transmission to humans. Abel Villa-Mancera, Pedro Molina-Mendoza, Karina Hernández-Guzmán, Jaime Olivares-Pérez, Jorge Sarracent-Pérez, and José Zumaquero-Ríos Copyright © 2016 Abel Villa-Mancera et al. All rights reserved. Management and Treatment of Human Lice Wed, 27 Jul 2016 13:04:06 +0000 Of the three lice (head, body, and pubic louse) that infest humans, the body louse is the species involved in epidemics of louse-borne typhus, trench fever, and relapsing fever, but all the three cause pediculosis. Their infestations occur today in many countries despite great efforts to maintain high standards of public health. In this review, literature searches were performed through PubMed, Medline, Google Scholar, and EBSCOhost, with key search words of “Pediculus humanus”, “lice infestation”, “pediculosis”, and “treatment”; and controlled clinical trials were viewed with great interest. Removing lice by hand or with a lice comb, heating infested clothing, and shaving the scalp were some of the oldest methods of controlling human lice. Despite the introduction of other resources including cresol, naphthalene, sulfur, mercury, vinegar, petroleum, and insecticides, the numbers of lice infestation cases and resistance have increased. To date, viable alternative treatments to replace insecticides have been developed experimentally in vitro. Today, the development of new treatment strategies such as symbiotic treatment and synergistic treatment (antibiotics + ivermectin) in vitro has proved effective and is promising. Here, we present an overview on managing and treating human lice and highlight new strategies to more effectively fight pediculosis and prevent resistance. Abdoul Karim Sangaré, Ogobara K. Doumbo, and Didier Raoult Copyright © 2016 Abdoul Karim Sangaré et al. All rights reserved. Phosphorylated and Nonphosphorylated PfMAP2 Are Localized in the Nucleus, Dependent on the Stage of Plasmodium falciparum Asexual Maturation Mon, 25 Jul 2016 14:29:16 +0000 Plasmodium falciparum mitogen-activated protein (MAP) kinases, a family of enzymes central to signal transduction processes including inflammatory responses, are a promising target for antimalarial drug development. Our study shows for the first time that the P. falciparum specific MAP kinase 2 (PfMAP2) is colocalized in the nucleus of all of the asexual erythrocytic stages of P. falciparum and is particularly elevated in its phosphorylated form. It was also discovered that PfMAP2 is expressed in its highest quantity during the early trophozoite (ring form) stage and significantly reduced in the mature trophozoite and schizont stages. Although the phosphorylated form of the kinase is always more prevalent, its ratio relative to the nonphosphorylated form remained constant irrespective of the parasites’ developmental stage. We have also shown that the TSH motif specifically renders PfMAP2 genetically divergent from the other plasmodial MAP kinase activation sites using Neighbour Joining analysis. Furthermore, TSH motif-specific designed antibody is crucial in determining the location of the expression of the PfMAP2 protein. However, by using immunoelectron microscopy, PPfMAP2 were detected ubiquitously in the parasitized erythrocytes. In summary, PfMAP2 may play a far more important role than previously thought and is a worthy candidate for research as an antimalarial. Farah Aida Dahalan, Hasidah Mohd Sidek, Mogana Das Murtey, Mohammed Noor Embi, Jamaiah Ibrahim, Lim Fei Tieng, Nurul Aiezzah Zakaria, and Noraishah Mydin Abdul-Aziz Copyright © 2016 Farah Aida Dahalan et al. All rights reserved. Geographical Distribution and New Situation of Leishmania Species after the Control of Cutaneous Leishmaniasis Foci in Errachidia Province, Morocco, in 2014 Tue, 28 Jun 2016 15:40:37 +0000 In Errachidia province, the incidence of cutaneous leishmaniasis (CL) has increased over the past decade and it was higher in 2010 (860.34 per 100,000 inhabitants), with 3445 cases. The number of cases declined sharply and decreased from 3445 cases in 2010 to 8 cases in 2014 following the control action plan interventions. The total of patients was diagnosed only on clinical basis and the lesions were considered caused by L. major. The epidemiological study was conducted between 2001 and 2014 and the molecular detection of CL was studied to identify the circulating parasite species in this province by using the ITS1-PCR-RFLP methods. In 2010, the molecular identification of 11 samples revealed the presence of L. major in the most affected circles: Goulmima, Er-Rissani, and Errachidia. In 2014 the molecular characterization of 7 among 8 cases reported in this year showed the presence of L. tropica in Errachidia circle. This is the first molecular identification of L. tropica in Errachidia province. The detection of this species after the intensified control measures strategies suggests that it was probably dissipated through the dominance of L. major. El Alem Mohamed Mahmoud, Sebti Faiza, Med Lemine, Chichaoui Smaine, El Bachir Adlaoui, Habbari Khalid, Sadak Abderrahim, and Fellah Hajiba Copyright © 2016 El Alem Mohamed Mahmoud et al. All rights reserved. Resistance Status to the Insecticides Temephos, Deltamethrin, and Diflubenzuron in Brazilian Aedes aegypti Populations Tue, 21 Jun 2016 14:20:46 +0000 Insecticides are still largely applied in public health to control disease vectors. In Brazil, organophosphates (OP) and pyrethroids (PY) are used against Aedes aegypti for years. Since 2009 Insect Growth Regulators (IGR) are also employed in the control of larvae. We quantified resistance to temephos (OP), deltamethrin (PY), and diflubenzuron (IGR) of A. aegypti samples from 12 municipalities distributed throughout the country, collected between 2010 and 2012. High levels of resistance to neurotoxic insecticides were detected in almost all populations: RR95 to temephos varied between 4.0 and 27.1; the lowest RR95 to deltamethrin was 13.1, and values higher than 70.0 were found. In contrast, all samples were susceptible to diflubenzuron (RR95 < 2.3). Biochemical tests performed with larvae and adults discarded the participation of acetylcholinesterase, the OP target, and confirmed involvement of the detoxifying enzymes esterases, mixed function oxidases, and glutathione-S-transferases. The results obtained were discussed taking into account the public chemical control component and the increase in the domestic use of insecticides during dengue epidemic seasons in the evaluated municipalities. Diogo Fernandes Bellinato, Priscila Fernandes Viana-Medeiros, Simone Costa Araújo, Ademir J. Martins, José Bento Pereira Lima, and Denise Valle Copyright © 2016 Diogo Fernandes Bellinato et al. All rights reserved. Immunopathological Features of Canine Myocarditis Associated with Leishmania infantum Infection Sun, 19 Jun 2016 11:05:02 +0000 Myocarditis associated with infectious diseases may occur in dogs, including those caused by the protozoa Neospora caninum, Trypanosoma cruzi, Babesia canis, and Hepatozoon canis. However, although cardiac disease due to Leishmania infection has also been documented, the immunopathological features of myocarditis have not been reported so far. The aim of this study was to examine the types of cellular infiltrates and expression of MHC classes I and II in myocardial samples obtained at necropsy from 15 dogs with an established intravitam diagnosis of visceral leishmaniasis. Pathological features of myocardium were characterized by hyaline degeneration of cardiomyocytes, necrosis, and infiltration of mononuclear inflammatory cells consisting of lymphocytes and macrophages, sometimes with perivascular pattern; fibrosis was also present in various degrees. Immunophenotyping of inflammatory cells was performed by immunohistochemistry on cryostat sections obtained from the heart of the infected dogs. The predominant leukocyte population was CD8+ with a fewer number of CD4+ cells. Many cardiomyocytes expressed MHC classes I and II on the sarcolemma. Leishmania amastigote forms were not detected within macrophages or any other cell of the examined samples. Our study provided evidence that myocarditis in canine visceral leishmaniasis might be related to immunological alterations associated with Leishmania infection. Alessandro Costagliola, Giuseppe Piegari, Iwona Otrocka-Domagala, Davide Ciccarelli, Valentina Iovane, Gaetano Oliva, Valeria Russo, Laura Rinaldi, Serenella Papparella, and Orlando Paciello Copyright © 2016 Alessandro Costagliola et al. All rights reserved. Histological Lesions and Cellular Response in the Skin of Alpine Chamois (Rupicapra r. rupicapra) Spontaneously Affected by Sarcoptic Mange Wed, 15 Jun 2016 11:39:53 +0000 Population dynamics of chamois (genus Rupicapra, subfamily Caprinae) can be influenced by infectious diseases epizootics, of which sarcoptic mange is probably the most severe in the Alpine chamois (Rupicapra rupicapra rupicapra). In this study, skin lesions and cellular inflammatory infiltrates were characterized in 44 Alpine chamois affected by sarcoptic mange. Dermal cellular responses were evaluated in comparison with chamois affected by trombiculosis and controls. In both sarcoptic mange and trombiculosis, a significantly increase of eosinophils, mast cells, T and B lymphocytes, and macrophages was detected. Moreover, in sarcoptic mange significant higher numbers of T lymphocytes and macrophages compared to trombiculosis were observed. Lesions in sarcoptic mange were classified in three grades, according to crusts thickness, correlated with mite counts. Grade 3 represented the most severe form with crust thickness more than 3.5 mm, high number of mites, and severe parakeratosis with diffuse bacteria. Evidence of immediate and delayed hypersensitivity was detected in all three forms associated with diffuse severe epidermal hyperplasia. In grade 3, a significant increase of B lymphocytes was evident compared to grades 1 and 2, while eosinophil counts were significantly higher than in grade 1, but lower than in grade 2 lesions. An involvement of nonprotective Th2 immune response could in part account for severe lesions of grade 3. Claudia Salvadori, Guido Rocchigiani, Camilla Lazzarotti, Nicoletta Formenti, Tiziana Trogu, Paolo Lanfranchi, Claudia Zanardello, Carlo Citterio, and Alessandro Poli Copyright © 2016 Claudia Salvadori et al. All rights reserved. Molecular Diagnosis and Identification of Leishmania Species in Jordan from Saved Dry Samples Wed, 15 Jun 2016 05:57:48 +0000 Diagnosis of the endemic cutaneous leishmaniasis (CL) in Jordan relies on patient clinical presentation and microscopic identification. Studies toward improved identification of the causative Leishmania species, especially in regions where multiple species exist, and the introduction of these techniques into medical diagnosis is paramount. This study looked at the current epidemiology of CL in Jordan. Clinically diagnosed 41 patients with CL were tested for the presence of Leishmania parasite using both Giemsa staining from skin scraps on glass slides and ITS1-PCR from samples blotted onto storage cards (NucleoCards®). Microscopically, 28 out of the 41 (68.3%) collected samples were positive for amastigotes, whereas the molecular ITS1-PCR amplification successfully identified 30 of the 41 samples (73.2%). Furthermore, PCR-RFLP analysis allowed species identification which is impossible microscopically. Of the 30 PCR positive samples, 28 were Leishmania major positive and the other two samples were Leishmania tropica. This indicates that L. major is the most prevalent species in Jordan and the two L. tropica cases originated from Syria indicating possible future L. tropica outbreaks. Diagnosis of CL based on clinical presentation only may falsely increase its prevalence. Although PCR is more sensitive, it is still not available in our medical laboratories in Jordan. Nawal Hijjawi, Kalil A. Kanani, Malak Rasheed, Manar Atoum, Mona Abdel-Dayem, and Mohammad R. Irhimeh Copyright © 2016 Nawal Hijjawi et al. All rights reserved. Biosensor for Hepatocellular Injury Corresponds to Experimental Scoring of Hepatosplenic Schistosomiasis in Mice Wed, 08 Jun 2016 12:21:42 +0000 Severe hepatosplenic injury of mansonian schistosomiasis is caused by Th2 mediated granulomatous response against parasite eggs entrapped within the periportal tissue. Subsequent fibrotic scarring and deformation/sclerosing of intrahepatic portal veins lead to portal hypertension, ascites, and oesophageal varices. The murine model of Schistosoma mansoni (S. mansoni) infection is suitable to establish the severe hepatosplenic injury of disease within a reasonable time scale for the development of novel antifibrotic or anti-infective strategies against S. mansoni infection. The drawback of the murine model is that the material prepared for complex analysis of egg burden, granuloma size, hepatic inflammation, and fibrosis is limited due to small amounts of liver tissue and blood samples. The objective of our study was the implementation of a macroscopic scoring system for mice livers to determine infection-related organ alterations of S. mansoni infection. In addition, an in vitro biosensor system based on the detection of hepatocellular injury in HepG2/C3A cells following incubation with serum of moderately (50 S. mansoni cercariae) and heavily (100 S. mansoni cercariae) infected mice affirmed the value of our scoring system. Therefore, our score represents a valuable tool in experimental schistosomiasis to assess severity of hepatosplenic schistosomiasis and reduce animal numbers by saving precious tissue samples. Martina Sombetzki, Nicole Koslowski, Sandra Doss, Micha Loebermann, Michael Trauner, Emil C. Reisinger, and Martin Sauer Copyright © 2016 Martina Sombetzki et al. All rights reserved. Anthelmintic Effect of Bacillus thuringiensis Strains against the Gill Fish Trematode Centrocestus formosanus Tue, 17 May 2016 12:21:43 +0000 Parasitic agents, such as helminths, are the most important biotic factors affecting aquaculture, and the fluke Centrocestus formosanus is considered to be highly pathogenic in various fish species. There have been efforts to control this parasite with chemical helminthicides, but these efforts have had unsuccessful results. We evaluated the anthelmintic effect of 37 strains of Bacillus thuringiensis against C. formosanus metacercariae in vitro using two concentrations of total protein, and only six strains produced high mortality. The virulence (CL50) on matacercariae of three strains was obtained: the GP308, GP526, and ME1 strains exhibited a LC50 of 146.2 μg/mL, 289.2 μg/mL, and 1721.9 μg/mL, respectively. Additionally, these six B. thuringiensis strains were evaluated against the cercariae of C. formosanus; the LC50 obtained from the GP526 strain with solubilized protein was 83.8 μg/mL, and it could be considered as an alternative control of the metacercariae and cercariae of this parasite in the productivity systems of ornamental fishes. Luis Javier Mendoza-Estrada, Víctor Manuel Hernández-Velázquez, Iván Arenas-Sosa, Fernando Iván Flores-Pérez, Jorge Morales-Montor, and Guadalupe Peña-Chora Copyright © 2016 Luis Javier Mendoza-Estrada et al. All rights reserved. West Nile Virus Circulation in Mosquitoes in Greece (2010–2013) Thu, 12 May 2016 13:06:07 +0000 Background of the Study. Following a large West Nile virus (WNV) epidemic in Northern Greece in 2010, an active mosquito surveillance system was implemented, for a 3-year period (2011, 2012, and 2013). Description of the Study Site and Methodology. Using mainly CO2 mosquito traps, mosquito collections were performed. Samples were pooled by date of collection, location, and species and examined for the presence of WNV. Results. Positive pools were detected in different areas of the country. In 2010, MIR and MLE values of 1.92 (95% CI: 0.00–4.57) and 2.30 (95% CI: 0.38–7.49) were calculated for the Serres Regional Unit in Central Macedonia Region. In 2011, the highest MIR value of 3.71(95% CI: 1.52–5.91) was recorded in the Regions of Central Greece and Thessaly. In 2012, MIR and MLE values for the whole country were 2.03 (95% CI: 1.73–2.33) and 2.15 (95% CI: 1.86–2.48), respectively, for Cx. pipiens. In 2013, in the Regional Unit of Attica, the one outbreak epicenter, MIR and MLE values for Cx. pipiens were 10.75 (95% CI: 7.52–13.99) and 15.76 (95% CI: 11.66–20.65), respectively. Significance of Results/Conclusions. The contribution of a mosquito-based surveillance system targeting WNV transmission is highlighted through the obtained data, as in most regions positive mosquito pools were detected prior to the date of symptom onset of human cases. Dissemination of the results on time to Public Health Authorities resulted in planning and application of public health interventions in local level. Eleni Patsoula, Annita Vakali, Georgios Balatsos, Danai Pervanidou, Stavroula Beleri, Nikolaos Tegos, Agoritsa Baka, Gregory Spanakos, Theano Georgakopoulou, Persefoni Tserkezou, Wim Van Bortel, Herve Zeller, Panagiotis Menounos, Jenny Kremastinou, and Christos Hadjichristodoulou Copyright © 2016 Eleni Patsoula et al. All rights reserved. Toxoplasma gondii and Neospora caninum in Free-Range Chickens in Henan Province of China Thu, 05 May 2016 11:49:22 +0000 Background. Chickens serve as an intermediate host for Toxoplasma gondii and Neospora caninum; infection of free-range (FR) chickens with these organisms is a useful indicator of soil and environmental contamination with oocysts. A total of 700 FR chicken serum samples and 300 heart samples were collected from Henan province from March to July 2015. Antibodies to T. gondii were found in 18.86% (132/700) of the chickens by modified agglutination test (cut-off 1 : 25), while 23.14% (162/700) were positive for N. caninum by indirect fluorescent antibody test (cut-off 1 : 25). T. gondii DNA was detected in the myocardium digestion liquids of 4/25 (16%) FR chickens. The PCR results of N. caninum DNA from FR chicken myocardium digestion liquids () were all negative. Attempts to isolate viable T. gondii were unsuccessful. The results showed that there were antibodies to T. gondii and N. caninum in FR chickens from Henan province. Accordingly, effective control of feces from cats and dogs and improved pets hygiene habits were needed. To the author’s knowledge, this is the first report of the presence of N. caninum antibody in chickens from China. Yongjie Feng, Yaoyao Lu, Yinghua Wang, Jing Liu, Longxian Zhang, and Yurong Yang Copyright © 2016 Yongjie Feng et al. All rights reserved. Leishmania infantum Genetic Diversity and Lutzomyia longipalpis Mitochondrial Haplotypes in Brazil Tue, 29 Mar 2016 12:35:07 +0000 Leishmania infantum is the etiological agent of visceral leishmaniasis (VL) in the Americas with domestic dogs being its major reservoir hosts. The main VL vector is the sandfly Lutzomyia longipalpis, while other Lutzomyia species may play a role in disease transmission. Although the genetic structure of L. infantum populations has been widely evaluated, only a few studies have addressed this subject coupled to the genetic structure of the respective sandfly vectors. In this study, we analyzed the population structure of L. infantum in three major VL endemic areas in Brazil and associated it with Lutzomyia longipalpis geographic structure. Paulo Eduardo Martins Ribolla, Letícia Tsieme Gushi, Maria do Socorro Pires e Cruz, Carlos Henrique Nery Costa, Dorcas Lamounier Costa, Manoel Sebastião da Costa Lima Júnior, Maria Elizabeth Moraes Cavalheiros Dorval, Alessandra Gutierrez de Oliveira, Mirella Ferreira da Cunha Santos, Vera Lúcia Fonseca Camargo-Neves, Carlos Magno Castello Branco Fortaleza, and Diego Peres Alonso Copyright © 2016 Paulo Eduardo Martins Ribolla et al. All rights reserved. Epidemiology of Plasmodium and Helminth Coinfection and Possible Reasons for Heterogeneity Tue, 22 Mar 2016 15:56:23 +0000 Understanding the impact of helminth infections on clinical malaria is useful for designing effective malaria control strategies. Plenty of epidemiological studies have been conducted to unravel the nature of interactions between Plasmodium and helminth infection. Careful broad summarization of the existing literature suggests that Schistosoma mansoni and hookworm infections may increase the risk of clinical malaria and associated morbidities, but Trichuris trichiura infection is not associated with the occurrence of clinical malaria and related outcomes. However, findings about effect of Ascaris lumbricoides and Schistosoma haematobium infection on clinical malaria are contradictory. Furthermore, the nature of relationship of helminth infection with severe malaria has also not been determined with certainty. This review summarizes the findings of epidemiological studies of Plasmodium and helminth coinfection, placing greater emphasis on the impact of the coinfection on malaria. Possible reasons for the heterogeneity of the findings on malaria and helminth coinfections are also discussed. Abraham Degarege and Berhanu Erko Copyright © 2016 Abraham Degarege and Berhanu Erko. All rights reserved. Tsetse Flies (Glossina) as Vectors of Human African Trypanosomiasis: A Review Mon, 29 Feb 2016 08:23:46 +0000 Human African Trypanosomiasis (HAT) transmitted by the tsetse fly continues to be a public health issue, despite more than a century of research. There are two types of the disease, the chronic gambiense and the acute rhodesiense-HAT. Fly abundance and distribution have been affected by changes in land-use patterns and climate. However, disease transmission still continues. Here, we review some aspects of HAT ecoepidemiology in the context of altered infestation patterns and maintenance of the transmission cycle as well as emerging options in disease and vector control. Florence Njeri Wamwiri and Robert Emojong Changasi Copyright © 2016 Florence Njeri Wamwiri and Robert Emojong Changasi. All rights reserved. Chemical Ecology of Parasitic Hymenoptera Mon, 22 Feb 2016 11:07:05 +0000 Giovanni Benelli, Kent M. Daane, Roxina Soler, and Johannes Stökl Copyright © 2016 Giovanni Benelli et al. All rights reserved. VOCs-Mediated Location of Olive Fly Larvae by the Braconid Parasitoid Psyttalia concolor: A Multivariate Comparison among VOC Bouquets from Three Olive Cultivars Thu, 18 Feb 2016 09:43:08 +0000 Herbivorous activity induces plant indirect defenses, as the emission of herbivorous-induced plant volatiles (HIPVs), which could be used by parasitoids for host location. Psyttalia concolor is a larval pupal endoparasitoid, attacking a number of tephritid flies including B. oleae. In this research, we investigated the olfactory cues routing host location behavior of P. concolor towards B. oleae larvae infesting three different olive cultivars. VOCs from infested and healthy fruits were identified using GC-MS analyses. In two-choice behavioral assays, P. concolor females preferred infested olive cues, which also evoked ovipositional probing by female wasps. GC-MS analysis showed qualitative and quantitative differences among volatiles emitted by infested and healthy olives. Volatile emissions were peculiar for each cultivar analyzed. Two putative HIPVs were detected in infested fruits, regardless of the cultivar, the monoterpene (E)-β-ocimene, and the sesquiterpene (E-E)-α-farnesene. Our study adds basic knowledge to the behavioral ecology of P. concolor. From an applied point of view, the field application of the above-mentioned VOCs may help to enhance effectiveness of biological control programs and parasitoid mass-rearing techniques. Giulia Giunti, Giovanni Benelli, Giuseppe Conte, Marcello Mele, Giovanni Caruso, Riccardo Gucci, Guido Flamini, and Angelo Canale Copyright © 2016 Giulia Giunti et al. All rights reserved. Newly Emerging Parasitic Threats for Human Health: National and International Trends Thu, 04 Feb 2016 08:00:11 +0000 Lidia Chomicz, David Bruce Conn, Jacek P. Szaflik, and Beata Szostakowska Copyright © 2016 Lidia Chomicz et al. All rights reserved. Development of a Murine Infection Model with Leishmania killicki, Responsible for Cutaneous Leishmaniosis in Algeria: Application in Pharmacology Tue, 02 Feb 2016 12:03:37 +0000 In Algeria, Leishmania infantum, Leishmania major, and Leishmania killicki (Leishmania tropica) are responsible for cutaneous leishmaniosis. We established a murine model of L. killicki infection to investigate its infective capacity, some immunophysiopathological aspects, and its suitability for pharmacological purposes. Following the injection of L. major or L. killicki metacyclic promastigotes in the ear dermis of BALB/c mice, the course of infection was followed. The infection with L. killicki caused slower lesion formation than with L. major. The presence of L. killicki or L. major DNA and parasites was detected in the ear dermis and in lymph nodes, spleen, and liver. Lesions induced by L. killicki were nonulcerative in their aspect, whereas those caused by L. major were highly ulcerative and necrotic, which matches well with the lesion phenotype reported in humans for L. killicki and L. major, respectively. The treatment of L. killicki lesions by injection of Glucantime® significantly reduced the lesion thickness and parasite burden. Ear dermal injection of BALB/c mice constitutes a model to study lesions physiopathology caused by L. killicki and presents interest for in vivo screening of new compounds against this pathogen, emerging in Algeria. Naouel Eddaikra, Ihcene Kherachi Djenad, Sihem Benbetka, Razika Benikhlef, Khatima Aït-Oudhia, Farida Moulti-Mati, Bruno Oury, Denis Sereno, and Zoubir Harrat Copyright © 2016 Naouel Eddaikra et al. All rights reserved. Electrophysiological and Behavioral Responses of Theocolax elegans (Westwood) (Hymenoptera: Pteromalidae) to Cereal Grain Volatiles Thu, 21 Jan 2016 13:33:08 +0000 Volatiles emitted by the host’s food would be the first signals used by parasitoids in the host location process and are thought to play an important role in host habitat location. In this study, the olfactory responses of Theocolax elegans (Westwood), a Pteromalid wasp that parasitizes immature stages of stored-product insect pests developing inside cereal or leguminous grains, to volatiles emitted by healthy wheat grains, their hexane extracts, and different doses of three individual compounds previously identified in cereal grain odors were investigated in Y-tube olfactometer and Petri dish arena behavioral bioassays and electroantennogram recordings. In Y-tube olfactometer bioassays, odors from healthy wheat grains and their hexane extracts were attractive to both sexes of T. elegans. Moreover, hexane extracts elicited arresting effects in Petri dish arena. The three synthetic compounds valeraldehyde, maltol, and vanillin elicited dose-dependent responses in both male and female adult wasps confirming the capability of the peripheral olfactory systems to perceive cereal volatiles. In behavioral bioassays, different doses of vanillin were significantly attractive to both sexes. Giacinto Salvatore Germinara, Antonio De Cristofaro, and Giuseppe Rotundo Copyright © 2016 Giacinto Salvatore Germinara et al. All rights reserved. Species Specificity of the Putative Male Antennal Aphrodisiac Pheromone in Leptopilina heterotoma, Leptopilina boulardi, and Leptopilina victoriae Tue, 29 Dec 2015 08:47:29 +0000 Male antennal aphrodisiac pheromones have been suggested to elicit female receptiveness in several parasitic Hymenoptera, including Leptopilina boulardi. None of the proposed pheromones, however, has been fully identified to date. It is also unknown whether these antennal pheromones are species specific, because the species specificity of mate recognition and courtship elicitation in Leptopilina prevented such experiments. In this study we present an experimental design that allows the investigation of the species specificity of the putative male aphrodisiac pheromone of L. heterotoma, L. boulardi, and L. victoriae. This is achieved by chemical manipulation of the odour profile of heterospecific females, so that males perceive them as conspecifics and show antennal courtship behaviour. Males courted the manipulated heterospecific females and antennal contact between the male and the female was observed. However, males elicited receptiveness only in conspecific females, never in the manipulated heterospecific females. Chemical analysis showed the presence of species specific unsaturated hydrocarbons on the antennae of males. Only trace amounts of these hydrocarbons are found on the antennae of females. Our results are an important step towards the understanding and identification of antennal pheromones of parasitic wasps. Ingmar Weiss, Joachim Ruther, and Johannes Stökl Copyright © 2015 Ingmar Weiss et al. All rights reserved. A Study on Health Seeking Behaviors of Patients of Post-Kala-Azar Dermal Leishmaniasis Tue, 15 Dec 2015 09:35:54 +0000 Post-Kala-Azar Dermal Leishmaniasis (PKDL) remains a major public health threat in Bangladesh. A cross-sectional study was carried out in Surya Kanta Kala azar Research Centre (SKKRC), Mymensingh, from January 2012 to July 2013 to evaluate the health seeking behaviour and the length of delay of PKDL management. The consecutive 200 diagnosed PKDL cases that got treatment in SKKRC hospital were subjected to evaluation. Most (98%) of the patients were not aware and had no knowledge about PKDL, though 87.5% had a history of history of Kala-azar treatment. Many patients reported first to village doctor (15.5%), the pharmacy shop (10%), or traditional health provider (7.5%) upon recognition of symptom. The time between the initial symptom recognition and first medical consultation (patient delay) ranged from 10 days to 4745 days (13 years) with a median of 373 days (mean: 696; IQR: 138 to 900 days). The time between first medical consultations to definite treatment (system delay) ranged from 0 days to 1971 days (5.4 years), with a median delay of 14 days (mean: 46.48; IQR: 7 to 44 days) that was reported in this study. Age, education, occupation, and residential status had significant association with patient delay (). Educational status, occupation, number of treatment providers, and first health care provider had a significant association with system delay (). Success in PKDL diagnosis and treatment requires specific behavior from patients and health care providers which facilitate those practices. Ariful Basher, Proggananda Nath, Shah Golam Nabi, Shahjada Selim, Md Fashiur Rahman, Satya Ranjan Sutradhar, Abul Faiz, Matiur Rahman Bhuiyan, Be-Nazir Ahmed, and Ridwanur Rahman Copyright © 2015 Ariful Basher et al. All rights reserved. Effects of Abiotic Factors on HIPV-Mediated Interactions between Plants and Parasitoids Tue, 15 Dec 2015 09:30:43 +0000 In contrast to constitutively emitted plant volatiles (PV), herbivore-induced plant volatiles (HIPV) are specifically emitted by plants when afflicted with herbivores. HIPV can be perceived by parasitoids and predators which parasitize or prey on the respective herbivores, including parasitic hymenoptera. HIPV act as signals and facilitate host/prey detection. They comprise a blend of compounds: main constituents are terpenoids and “green leaf volatiles.” Constitutive emission of PV is well known to be influenced by abiotic factors like temperature, light intensity, water, and nutrient availability. HIPV share biosynthetic pathways with constitutively emitted PV and might therefore likewise be affected by abiotic conditions. However, the effects of abiotic factors on HIPV-mediated biotic interactions have received only limited attention to date. HIPV being influenced by the plant’s growing conditions could have major implications for pest management. Quantitative and qualitative changes in HIPV blends may improve or impair biocontrol. Enhanced emission of HIPV may attract a larger number of natural enemies. Reduced emission rates or altered compositions, however, may render blends imperceptible to parasitoides and predators. Predicting the outcome of these changes is highly important for food production and for ecosystems affected by global climate change. Christine Becker, Nicolas Desneux, Lucie Monticelli, Xavier Fernandez, Thomas Michel, and Anne-Violette Lavoir Copyright © 2015 Christine Becker et al. All rights reserved. Molecular Detection of Theileria spp. in Livestock on Five Caribbean Islands Wed, 09 Dec 2015 06:43:37 +0000 Theileria spp. are tick-transmitted, intracellular apicomplexan protozoan parasites infecting a wide range of animals. As there is very limited information on the prevalence of Theileria spp. in the Caribbean we used the recently described genus-specific pan-Theileria FRET-qPCR to identify infected animals in the region and a standard 18S rRNA gene PCR and sequencing to determine the species involved. We found Theileria spp. in 9% of the convenience samples of animals () studied from five Caribbean islands. Donkeys (20.0%: 5/25) were most commonly infected, followed by sheep (17.4%, 25/144), cattle (6.8%; 22/325), goats (5.0%; 12/238), and horses (5.0%; 1/20). Six species of Theileria were identified: T. equi (donkeys, cattle, goats, and sheep), Theileria sp. OT3 (sheep and goats), Theileria sp. NG-2013a (cattle), Theileria sp. YW-2014 (donkeys), Theileria sp. B15a (goats), and Babesia vulpes or a closely related organism (sheep and goats). Only T. equi has been previously reported in the Caribbean. Our findings expand the known host ranges of Theileria spp. and the known distribution of the organisms around the world. Jilei Zhang, Patrick Kelly, Jing Li, Chuanling Xu, and Chengming Wang Copyright © 2015 Jilei Zhang et al. All rights reserved. Emerging Threats for Human Health in Poland: Pathogenic Isolates from Drug Resistant Acanthamoeba Keratitis Monitored in terms of Their In Vitro Dynamics and Temperature Adaptability Mon, 23 Nov 2015 11:50:55 +0000 Amphizoic amoebae generate a serious human health threat due to their pathogenic potential as facultative parasites, causative agents of vision-threatening Acanthamoeba keratitis (AK). Recently, AK incidences have been reported with increasing frequency worldwide, particularly in contact lens wearers. In our study, severe cases of AK in Poland and respective pathogenic isolates were assessed at clinical, morphological, and molecular levels. Misdiagnoses and the unsuccessful treatment in other ophthalmic units delayed suitable therapy, and resistance to applied chemicals resulted in severe courses and treatment difficulties. Molecular assessment indicated that all sequenced pathogenic corneal isolates deriving from Polish patients with AK examined by us showed 98–100% homology with Acanthamoeba genotype T4, the most prevalent genotype in this human ocular infection worldwide. In vitro assays revealed that the pathogenic strains are able to grow at elevated temperature and have a wide adaptive capability. This study is our subsequent in vitro investigation on pathogenic Acanthamoeba strains of AK originating from Polish patients. Further investigations designed to foster a better understanding of the factors leading to an increase of AK observed in the past years in Poland may help to prevent or at least better cope with future cases. Lidia Chomicz, David Bruce Conn, Marcin Padzik, Jacek P. Szaflik, Julia Walochnik, Paweł J. Zawadzki, Witold Pawłowski, and Monika Dybicz Copyright © 2015 Lidia Chomicz et al. All rights reserved. Challenges for Diagnosis of Malaria and Neglected Tropical Diseases in Elimination Settings Thu, 05 Nov 2015 08:29:05 +0000 Stephan Karl, Malcolm K. Jones, Lucía Gutiérrez, Brioni Moore, Eline Kattenberg, and Marcus Lacerda Copyright © 2015 Stephan Karl et al. All rights reserved. Sequence Variation in HSP40 Gene among 16 Toxoplasma gondii Isolates from Different Hosts and Geographical Locations Tue, 03 Nov 2015 06:14:21 +0000 Toxoplasma gondii with worldwide distribution has received substantial medical and scientific attentions as it causes serious clinical and veterinary problems especially for pregnant women and immunocompromised patients. Heat shock protein 40 (HSP40) plays a variety of essential roles in the pathogenesis of this protozoan parasite. In order to detail the genetic diversity of HSP40 gene, 16 T. gondii strains from different hosts and geographical locations were used in this study. Our results showed that HSP40 sequence of the examined strains was between 6621 bp and 6644 bp in length, and their A+T content was from 48.54% to 48.80%. Furthermore, sequence analysis presented 195 nucleotide mutation positions (0.12%–1.14%) including 29 positions in CDS (0.02%–0.12%) compared with T. gondii ME49 strain (ToxoDB: TGME49_265310). Phylogenetic assay revealed that T. gondii strains representing three classical genotypes (Types I, II, and III) were completely separated into different clusters by maximum parsimony (MP) method, but Type II and ToxoDB#9 strains were grouped into the same cluster. These results suggested that HSP40 gene is not a suitable marker for T. gondii population genetic research, though three classical genotypes of T. gondii could be differentiated by restriction enzymes MscI and EarI existing in amplicon C. Zhong-Yuan Li, Jing Lu, Dong-Hui Zhou, Jia Chen, and Xing-Quan Zhu Copyright © 2015 Zhong-Yuan Li et al. All rights reserved. Syncytiotrophoblast Functions and Fetal Growth Restriction during Placental Malaria: Updates and Implication for Future Interventions Mon, 26 Oct 2015 08:25:49 +0000 Syncytiotrophoblast lines the intervillous space of the placenta and plays important roles in fetus growth throughout gestation. However, perturbations at the maternal-fetal interface during placental malaria may possibly alter the physiological functions of syncytiotrophoblast and therefore growth and development of the embryo in utero. An understanding of the influence of placental malaria on syncytiotrophoblast function is paramount in developing novel interventions for the control of placental pathology associated with placental malaria. In this review, we discuss how malaria changes syncytiotrophoblast function as evidenced from human, animal, and in vitro studies and, further, how dysregulation of syncytiotrophoblast function may impact fetal growth in utero. We also formulate a hypothesis, stemming from epidemiological observations, that nutrition may override pathogenesis of placental malaria-associated-fetal growth restriction. We therefore recommend studies on nutrition-based-interventional approaches for high placental malaria-risk women in endemic areas. More investigations on the role of nutrition on placental malaria pathogenesis are needed. Winifrida B. Kidima Copyright © 2015 Winifrida B. Kidima. All rights reserved. Revisiting the Posttherapeutic Cure Criterion in Chagas Disease: Time for New Methods, More Questions, Doubts, and Polemics or Time to Change Old Concepts? Sun, 25 Oct 2015 11:40:13 +0000 One of the most relevant issues beyond the effectiveness of etiological treatment of Chagas disease is the lack of consensual/feasible tools to identify and certify the definitive parasitological cure. Several methods of distinct natures (parasitological, serological, and molecular) have been continuously proposed and novel perspectives are currently under investigation. Although the simultaneous use of distinct tests may offer better contributions and advances, it also leads to controversies of interpretation, with lack of mutual consent of cure criterion amongst researchers and physicians. In fact, when distinct host compartments (blood/tissues) are evaluated and explored, novel questions may arise due to the nature and sensitivity limit of each test. This short analytical review intends to present a chronological and critical overview and discuss the state-of-the-art distinct devices available for posttherapeutic cure assessment in Chagas disease, their contributions, meanings, and interpretation, aiming to point out the major gaps and propose novel insight for future perspectives of posttherapeutic management of Chagas disease patients. Marta de Lana and Olindo Assis Martins-Filho Copyright © 2015 Marta de Lana and Olindo Assis Martins-Filho. All rights reserved. Species Composition and Diversity of Malaria Vector Breeding Habitats in Trincomalee District of Sri Lanka Sun, 25 Oct 2015 11:37:23 +0000 Introduction. Mosquito larval ecology is important in determining larval densities and species assemblage. This in turn influences malaria transmission in an area. Therefore, understanding larval habitat ecology is important in designing malaria control programs. Method. Larval surveys were conducted in 20 localities under five sentinel sites (Padavisiripura, Gomarankadawala, Thoppur, Mollipothana, and Ichchallampaththu) in Trincomalee District, Eastern Province of Sri Lanka, between June 2010 and July 2013. The relationship between seven abiotic variables (temperature, pH, conductivity, Total Dissolved Solid (TDS), turbidity, Dissolved Oxygen (DO), and salinity) was measured. Results. A total of 21,347 anophelines were recorded representing 15 species. Anopheles subpictus 24.72% (5,278/21,347) was the predominant species, followed by 24.67% (5,267/21,347) of An. nigerrimus and 14.56% (3,109/21,347) of An. peditaeniatus. A total of 9,430 breeding habitats under twenty-one categories were identified. An. culcicifacies was noted to be highest from built wells (20.5%) with high salinity (1102.3 ± 81.8 mg/L), followed by waste water collections (20.2%) having low DO levels (2.85 ± 0.03 mg/L) and high TDS (1,654 ± 140 mg/L). Conclusion. This study opens an avenue to explore new breeding habitats of malaria vectors in the country and reemphasizes the requirement of conducting entomological surveillance to detect potential transmission of malaria in Sri Lanka under the current malaria elimination programme. Nayana Gunathilaka, Wimaladharma Abeyewickreme, Menaka Hapugoda, and Rajitha Wickremasinghe Copyright © 2015 Nayana Gunathilaka et al. All rights reserved. Importation and Transmission of Parasitic and Other Infectious Diseases Associated with International Adoptees and Refugees Immigrating into the United States of America Sun, 25 Oct 2015 11:12:14 +0000 Each year, hundreds of millions of people travel across international borders or even oceans, and up to 230 million may remain for long periods. Among these, 3–5 million settle permanently in their new homes, with about 1 million migrating permanently to the United States of America. This may result in transport of parasites and other pathogens, which might become established, infecting individuals in the new location. Beyond concern of disease spread, the health of migrants is of concern since the rigors, circumstances, and living conditions surrounding migrations may increase the vulnerability of migrants to infections. International adoptees and refugees are a small subset of these migrants but are of special significance inasmuch as adoptees may be more vulnerable to infection due to their immature immune status, and refugees may be more vulnerable due to substandard living conditions. Both originate from diverse regions, but often from environments of low hygiene and health care standards. This review examines recent examples of infections reported from adoptees and refugees entering the USA through 2010, highlighting the most common origin countries and the diseases most frequently involved, including Chagas disease, Balamuthia amebic meningoencephalitis, giardiasis, microsporidiosis, hepatitis, measles, pertussis, tuberculosis, malaria, intestinal helminths, and syphilis. Jordan Smith Darr and David Bruce Conn Copyright © 2015 Jordan Smith Darr and David Bruce Conn. All rights reserved. Identification and Biological Characterization of Leishmania (Viannia) guyanensis Isolated from a Patient with Tegumentary Leishmaniasis in Goiás, a Nonendemic Area for This Species in Brazil Sun, 25 Oct 2015 09:41:38 +0000 The aim of this study was to characterize clinical field isolates of Leishmania spp. obtained from patients with American Tegumentary Leishmaniasis (ATL) who live in Goiás state, Brazil. The presumed areas of infection were in Goiás, Tocantins, and Pará states. Three isolates of parasites were identified as L. (Viannia) braziliensis and one as L. (V.) guyanensis. The in vitro growth profiles were found to be similar for all parasites. Nevertheless, in C57BL/6 mice, L. (V.) guyanensis infection was better controlled than L. (V.) braziliensis. Yet in C57BL/6 mice deficient in interferon gamma, L. (V.) guyanensis lesions developed faster than those caused by L. (V.) braziliensis isolates. In BALB/c mice, the development of lesions was similar for isolates from both species; however, on the 11th week of infection, amastigotes could not be observed in macrophages from L. (V.) guyanensis-infected mice. Thus, L. (V.) guyanensis can be circulating in Goiás, a state where autochthonous cases of this species had not yet been reported. Considering the difficulties to differentiate L. (V.) guyanensis from L. (V.) braziliensis at the molecular, morphological, and clinical (human and murine models) levels, the presence of L. (V.) guyanensis infections is possibly underestimated in several regions of Brazil. Alause da Silva Pires, Arissa Felipe Borges, Adriano Cappellazzo Coelho, Miriam Leandro Dorta, Ruy de Souza Lino Junior, Ledice Inacia de Araújo Pereira, Sebastião Alves Pinto, Milton Adriano Pelli de Oliveira, Grazzielle Guimarães de Matos, Ises A. Abrahamsohn, Silvia Reni B. Uliana, Glória Maria Collet de Araújo Lima, and Fátima Ribeiro-Dias Copyright © 2015 Alause da Silva Pires et al. All rights reserved. Sexy Mouth Odour? Male Oral Gland Pheromone in the Grain Beetle Parasitoid Lariophagus distinguendus (Förster) (Hymenoptera: Pteromalidae) Thu, 22 Oct 2015 11:48:44 +0000 Throughout the animal kingdom, sexual pheromones are used for the attraction of mates and as courtship signals but also enable sexual isolation between species. In the parasitic wasp Lariophagus distinguendus, male courtship behaviour consisting of wing fanning, antennal stroking of the female antenna, and head nodding stimulates female receptivity leading to copulation. Recently L. distinguendus was reported to consist of two different lineages, which are sexually isolated because males fail to elicit receptivity in foreign females. It is unclear, however, which part of the courtship behaviour triggers female receptivity and therefore could be a mechanism causing sexual isolation. Here we show that in L. distinguendus a nonvolatile male oral pheromone is essential to release the female receptivity signal. In contrast, male wing fanning and antennal contact play a minor role. Additionally, the composition of the oral pheromone depends on the developmental host and females learn the composition upon emergence from the host substrate. These results will enable more detailed work on oral sexual pheromones to answer the question of how they are involved in the speciation process of L. distinguendus and other parasitoid species, for a better understanding of the huge biodiversity in this group. Kerstin König, Lucy Seeger, and Johannes L. M. Steidle Copyright © 2015 Kerstin König et al. All rights reserved. Prevalence of Malaria Infection and Risk Factors Associated with Anaemia among Pregnant Women in Semiurban Community of Hazaribag, Jharkhand, India Tue, 13 Oct 2015 08:39:51 +0000 The escalating burden, pathogenesis, and clinical sequel of malaria during pregnancy have combinatorial adverse impact on both mother and foetus that further perplexed the situation of diagnosis, treatment, and prevention. This prompted us to evaluate the status of population at risk of MIP in Hazaribag, Jharkhand, India. Cross-sectional study was conducted over a year at Sadar Hospital, Hazaribag. Malaria was screened using blood smear and/or RDT. Anaemia was defined as haemoglobin concentration. Pretested questionnaires were used to gather sociodemographic, clinical, and obstetrical data. The prevalence of MIP was 5.4% and 4.3% at ANC and DU, and 13.2% malaria was in women without pregnancy. Interestingly, majority were asymptomatically infected with P. vivax (over 85%) at ANC and DU. Peripheral parasitemia was significantly associated with fever within past week, rural origin of subjects, and first/second pregnancies in multivariate analysis, with the highest risk factor associated with fever followed by rural residence. Strikingly in cohort, anaemia was prevalent in 86% at ANC as compared to 72% at DU, whereas severe anaemia was 13.6% and 7.8% at ANC and DU. Even more anaemia prevalence was observed in MIP group (88% and 89% at ANC and DU), whereas severe anaemia was 23% and 21%, respectively. In view of observed impact of anaemia, parasitemia and asymptomatic infection of P. vivax during pregnancy and delivery suggest prompt diagnosis regardless of symptoms and comprehensive drug regime should be offered to pregnant women in association with existing measures in clinical spectrum of MIP, delivery, and its outcome. Mohammad Sohail, Shayan Shakeel, Shweta Kumari, Aakanksha Bharti, Faisal Zahid, Shadab Anwar, Krishn Pratap Singh, Mazahirul Islam, Ajay Kumar Sharma, Sneh Lata, Vahab Ali, Tridibes Adak, Pradeep Das, and Mohammad Raziuddin Copyright © 2015 Mohammad Sohail et al. All rights reserved. Anagrus breviphragma Soyka Short Distance Search Stimuli Mon, 12 Oct 2015 08:04:39 +0000 Anagrus breviphragma Soyka (Hymenoptera: Mymaridae) successfully parasitises eggs of Cicadella viridis (L.) (Homoptera: Cicadellidae), embedded in vegetal tissues, suggesting the idea of possible chemical and physical cues, revealing the eggs presence. In this research, three treatments were considered in order to establish which types of cue are involved: eggs extracted from leaf, used as a control, eggs extracted from leaf and cleaned in water and ethanol, used to evaluate the presence of chemicals soluble in polar solvents, and eggs extracted from leaf and covered with Parafilm (M), used to avoid physical stimuli due to the bump on the leaf surface. The results show that eggs covered with Parafilm present a higher number of parasitised eggs and a lower probing starting time with respect to eggs washed with polar solvents or eggs extracted and untreated, both when the treatments were singly tested or when offered in sequence, independently of the treatment position. These results suggest that the exploited stimuli are not physical due to the bump but chemicals that can spread in the Parafilm, circulating the signal on the whole surface, and that the stimuli that elicit probing and oviposition are not subjected to learning. Elisabetta Chiappini, Alessia Berzolla, and Annalisa Oppo Copyright © 2015 Elisabetta Chiappini et al. All rights reserved. Diagnostic Options and Challenges for Dengue and Chikungunya Viruses Mon, 05 Oct 2015 07:59:30 +0000 Dengue virus (DENV) and Chikungunya virus (CHIKV) are arboviruses that share the same Aedes mosquito vectors and thus overlap in their endemic areas. These two viruses also cause similar clinical presentations, especially in the initial stages of infection, with neither virus possessing any specific distinguishing clinical features. Because the outcomes and management strategies for these two viruses are vastly different, early and accurate diagnosis is imperative. Diagnosis is also important for surveillance, outbreak control, and research related to vaccine and drug development. Available diagnostic tests are aimed at detection of the virus, its antigenic components, or the host immune antibody response. In this review, we describe the recent progress and continued challenges related to the diagnosis of DENV and CHIKV infections. Stacey K. Mardekian and Amity L. Roberts Copyright © 2015 Stacey K. Mardekian and Amity L. Roberts. All rights reserved. Overview of the Diagnostic Methods Used in the Field for Human African Trypanosomiasis: What Could Change in the Next Years? Sun, 04 Oct 2015 13:26:30 +0000 Sleeping sickness is a parasitic infection caused by two species of trypanosomes (Trypanosoma brucei gambiense and rhodesiense), transmitted by the tsetse fly. The disease eventually affects the central nervous system, resulting in severe neurological symptoms. Without treatment, death is inevitable. During the first stage of the disease, infected patients are mildly symptomatic and early detection of infection allows safer treatment (administered on an outpatient basis) which can avoid death; routine screening of the exposed population is necessary, especially in areas of high endemicity. The current therapeutic treatment of this disease, especially in stage 2, can cause complications and requires a clinical surveillance for several days. A good stage diagnosis of the disease is the cornerstone for delivering the adequate treatment. The task faced by the medical personnel is further complicated by the lack of support from local health infrastructure, which is at best weak, but often nonexistent. Therefore it is crucial to look for new more efficient technics for the diagnosis of stage which are also best suited to use in the field, in areas not possessing high-level health facilities. This review, after an overview of the disease, summarizes the current diagnosis procedures and presents the advances in the field. Julien Bonnet, Clotilde Boudot, and Bertrand Courtioux Copyright © 2015 Julien Bonnet et al. All rights reserved. Loop Mediated Isothermal Amplification for Detection of Trypanosoma brucei gambiense in Urine and Saliva Samples in Nonhuman Primate Model Sun, 04 Oct 2015 10:23:12 +0000 Human African trypanosomiasis (HAT) is a vector-borne parasitic zoonotic disease. The disease caused by Trypanosoma brucei gambiense is the most prevalent in Africa. Early diagnosis is hampered by lack of sensitive diagnostic techniques. This study explored the potential of loop mediated isothermal amplification (LAMP) and polymerase chain reaction (PCR) in the detection of T. b. gambiense infection in a vervet monkey HAT model. Six vervet monkeys were experimentally infected with T. b. gambiense IL3253 and monitored for 180 days after infection. Parasitaemia was scored daily. Blood, cerebrospinal fluid (CSF), saliva, and urine samples were collected weekly. PCR and LAMP were performed on serum, CSF, saliva, and urine samples. The detection by LAMP was significantly higher than that of parasitological methods and PCR in all the samples. The performance of LAMP varied between the samples and was better in serum followed by saliva and then urine samples. In the saliva samples, LAMP had 100% detection between 21 and 77 dpi, whereas in urine the detection it was slightly lower, but there was over 80% detection between 28 and 91 dpi. However, LAMP could not detect trypanosomes in either saliva or urine after 140 and 126 dpi, respectively. The findings of this study emphasize the importance of LAMP in diagnosis of HAT using saliva and urine samples. Maina Ngotho, John Maina Kagira, Beatrice Muthoni Gachie, Simon Muturi Karanja, Maxwell Wambua Waema, Dawn Nyawira Maranga, and Naomi Wangari Maina Copyright © 2015 Maina Ngotho et al. All rights reserved. Comparative Study of the Accuracy of Different Techniques for the Laboratory Diagnosis of Schistosomiasis Mansoni in Areas of Low Endemicity in Barra Mansa City, Rio de Janeiro State, Brazil Sun, 04 Oct 2015 09:18:58 +0000 Schistosomiasis constitutes a major public health problem, with an estimated 200 million people infected worldwide. Many areas of Brazil show low endemicity of schistosomiasis, and the current standard parasitological techniques are not sufficiently sensitive to detect the low-level helminth infections common in areas of low endemicity (ALEs). This study compared the Kato-Katz (KK); Hoffman, Pons, and Janer (HH); enzyme-linked immunosorbent assay- (ELISA-) IgG and ELISA-IgM; indirect immunofluorescence technique (IFT-IgM); and qPCR techniques for schistosomiasis detection in serum and fecal samples, using the circumoval precipitin test (COPT) as reference. An epidemiological survey was conducted in a randomized sample of residents from five neighborhoods of Barra Mansa, RJ, with 610 fecal and 612 serum samples. ELISA-IgM (21.4%) showed the highest positivity and HH and KK techniques were the least sensitive (0.8%). All techniques except qPCR-serum showed high accuracy (82–95.5%), differed significantly from COPT in positivity , and showed poor agreement with COPT. Medium agreement was seen with ELISA-IgG (Kappa = 0.377) and IFA (Kappa = 0.347). Parasitological techniques showed much lower positivity rates than those by other techniques. We suggest the possibility of using a combination of laboratory tools for the diagnosis of schistosomiasis in ALEs. Maria Cristina Carvalho Espírito-Santo, Mónica Viviana Alvarado-Mora, Pedro Luiz Silva Pinto, Maria Carmen Arroyo Sanchez, Emmanuel Dias-Neto, Vera Lúcia Pagliusi Castilho, Elenice Messias do Nascimento Gonçalves, Pedro Paulo Chieffi, Expedito José de Albuquerque Luna, João Renato Rebello Pinho, Flair José Carrilho, and Ronaldo Cesar Borges Gryschek Copyright © 2015 Maria Cristina Carvalho Espírito-Santo et al. All rights reserved. Serological Diagnosis and Follow-Up of Human Cystic Echinococcosis: A New Hope for the Future? Sun, 04 Oct 2015 09:15:24 +0000 Cystic echinococcosis (CE) is an important helminthic zoonotic disease caused by the Echinococcus granulosus complex. In humans, CE is a chronic disease driven by the growth of echinococcal cysts in different organs. Prognosis of this disease depends on multiple factors, including location, number, size, and stage of the cysts, making CE a disease of complex management. CE is usually asymptomatic for years and attracts limited attention from funding organizations and health authorities. For this reason, only experts’ recommendations are available but no evidence-based conclusions have been drawn for CE clinical management. One of those pitfalls refers to the lack of evidence to support the use of serological tools for the diagnosis and follow-up of CE patients. In this respect, crude antigens are used to detect specific antibodies in patients, giving rise to false positive results. The advent of molecular techniques allowing the production of recombinant proteins has provided a number of candidate antigens that could overcome the problems associated with the use of crude parasite extracts in the serological assays. In this review, we present the last advances in this field, proposing the use of serology to support cyst stage-specific diagnosis and follow-up. Raúl Manzano-Román, Carlos Sánchez-Ovejero, Ana Hernández-González, Adriano Casulli, and Mar Siles-Lucas Copyright © 2015 Raúl Manzano-Román et al. All rights reserved. Cases of Echinococcus granulosus Sensu Stricto Isolated from Polish Patients: Imported or Indigenous? Wed, 30 Sep 2015 16:36:42 +0000 The cases of nine Polish patients with diagnosed cystic echinococcosis (CE) were examined. A total of nine isolates obtained postoperatively were investigated using PCR and sequencing. The mitochondrial region of nad1 gene was amplified. This PCR and sequencing analysis revealed the presence of Echinococcus canadensis G7 in seven patients and E. granulosus G1 in two patients. These data demonstrate that E. canadensis is the predominant causative agent of human cystic echinococcosis in Poland. E. granulosus G1 detection in Polish patients suggests that the parasite was imported; however it does not exclude the possibility that these cases could have been of Polish origin. Monika Dybicz, Piotr Karol Borkowski, Julia Dąbrowska, and Lidia Chomicz Copyright © 2015 Monika Dybicz et al. All rights reserved. Imported Malaria in the Material of the Institute of Maritime and Tropical Medicine: A Review of 82 Patients in the Years 2002–2014 Wed, 16 Sep 2015 07:56:57 +0000 Malaria is, along with tuberculosis and HIV/AIDS, one of the three most dangerous infectious diseases in the world. In the absence of native cases since 1963, malaria has remained in Poland an exclusively imported disease, mainly occurring in people travelling to tropical and subtropical areas for professional reasons. The aim of this study was the epidemiological and clinical analysis of 82 patients admitted to the University Center for Maritime and Tropical Medicine (UCMTM), Gdynia, Poland, with a diagnosis of malaria between 2002 and 2014. The “typical” patient with malaria was male, middle-aged, returned from Africa within the preceding 4 weeks, had not used appropriate chemoprophylaxis, and had not applied nonpharmacological methods of prophylaxis, except for window insect screens. P. falciparum was the most frequent species. The most common symptoms included fever, shivers and intensive sweating, thrombocytopenia, elevated creatinine, LDH, D-dimers and CRP, hepatomegaly, and splenomegaly. Within the analyzed group, severe malaria according to WHO standards was diagnosed in 20.7% of patients. Our report presents analysis of the largest series of patients treated for imported malaria in Poland. Anna Kuna, Michal Gajewski, Beata Szostakowska, Waclaw L. Nahorski, Przemyslaw Myjak, and Joanna Stanczak Copyright © 2015 Anna Kuna et al. All rights reserved. A Multiplex PCR for Simultaneous Detection of Three Zoonotic Parasites Ancylostoma ceylanicum, A. caninum, and Giardia lamblia Assemblage A Mon, 31 Aug 2015 11:58:17 +0000 Ancylostoma ceylanicum, A. caninum, and Giardia lamblia assemblage A are common intestinal parasites of dogs and cats; they can also infect humans, causing parasitic zoonoses. In this study, a multiplex PCR method was developed for simultaneous identification and detection of those three zoonotic parasites. Three pairs of specific primers were designed based on ITS sequence of A. ceylanicum and A. caninum and TPI gene of G. lamblia available in the GenBank. The multiplex PCR reaction system was established by optimizing the reaction condition, and a series of tests on the sensitivity, specificity, and clinical application were also conducted. Results showed that three target fragments were amplified specifically; the detection limit was 10 eggs for both A. ceylanicum and A. caninum, 72 pg DNA for G. lamblia. Of 112 clinical fecal samples, 34.8% and 17.8% samples were positive for A. caninum and A. ceylanicum, respectively, while only 2.7% samples were positive for G. lamblia assemblage A. It is concluded that the established multiplex PCR assay is a convenient, rapid, cost-effective, and high-efficiency method for molecular detection and epidemiological investigation of three zoonotic parasites. Wei Hu, Sheng Wu, Xingang Yu, Auwalu Yusuf Abullahi, Meiran Song, Liping Tan, Zhen Wang, Biao Jiang, and Guoqing Li Copyright © 2015 Wei Hu et al. All rights reserved. An Integrated Approach Using Spatial Analysis to Study the Risk Factors for Leishmaniasis in Area of Recent Transmission Wed, 01 Jul 2015 12:07:00 +0000 Some epidemiological aspects of leishmaniasis in the municipality of Formiga, Brazil, an important touristic site, were evaluated. Those included phlebotomine sand fly vectors, canine infection, and geoprocessing analysis for determining critical transmission areas. Sand flies (224 insects) belonging to ten different species were captured. The most captured species included Lutzomyia longipalpis (35.3%), Lutzomyia cortelezzii (33.5%), and Lutzomyia whitmani (18.3%). A significant correlation between sand fly densities and climatic conditions was detected. Serological diagnosis (DPP and ELISA) was performed in 570 dogs indicating a prevalence of 5.8%. After sequencing the main species circulating in the area were Leishmania infantum and Leishmania braziliensis. Spatial analysis demonstrated that vegetation and hydrography may be related to sand fly distribution and infected dogs. The municipality of Formiga has proven leishmaniasis vectors and infected dogs indicating the circulation of the parasite in the city. Correlation of those data with environmental and human cases has identified the critical areas for control interventions (south, northeast, and northwest). In conclusion, there is current transmission of visceral and canine human cases and the city is on the risk for the appearance of cutaneous cases. Júlia Alves Menezes, Eduardo de Castro Ferreira, José Dilermando Andrade-Filho, Alessandra Mara de Sousa, Mayron Henrique Gomes Morais, Ana Maria Sampaio Rocha, George Luis Lins Machado-Coelho, Fernanda Pinheiro Lima, Ana Paula Madureira, Tânia Cristina Garcia, Christian Resende Freitas, Rodrigo Pedro Soares, and Carina Margonari Copyright © 2015 Júlia Alves Menezes et al. All rights reserved. The First Survey of Forensically Important Entomofauna Collected from Medicolegal Autopsies in South Korea Mon, 22 Jun 2015 09:02:42 +0000 Forensic entomology applies insect evidence to legal problems such as the estimation of minimum postmortem interval (mPMI). For this purpose, knowledge of the insect fauna that are attracted to human cadavers in each geographic region is a prerequisite. Despite many studies investigating the insect fauna attracted to meat, there has been no survey of the entomofauna on human cadavers in the East Asian temperate climate zone, particularly in Korea. Therefore, this study reports the entomofauna collected from medicolegal autopsies in northeastern Seoul and its suburbs. Insect samples were collected from 35 medicolegal autopsies in 2010, 2011, and 2013. Molecular and morphological methods were utilized for taxonomic identification. Among 1398 individual samples belonging to 3 orders, 13 families, 18 genera, and 32 species, the dominant family and species were Calliphoridae and Lucilia sericata, respectively. Despite its limited scale, this study provides a snapshot of the general entomofauna that are attracted to human cadavers in this region. Sang Eon Shin, Hyun Ju Lee, Ji Hye Park, Kwang Soo Ko, Yu-Hoon Kim, Kyung Ryoul Kim, and Seong Hwan Park Copyright © 2015 Sang Eon Shin et al. All rights reserved. Molecular Chaperones of Leishmania: Central Players in Many Stress-Related and -Unrelated Physiological Processes Thu, 18 Jun 2015 08:33:07 +0000 Molecular chaperones are key components in the maintenance of cellular homeostasis and survival, not only during stress but also under optimal growth conditions. Folding of nascent polypeptides is supported by molecular chaperones, which avoid the formation of aggregates by preventing nonspecific interactions and aid, when necessary, the translocation of proteins to their correct intracellular localization. Furthermore, when proteins are damaged, molecular chaperones may also facilitate their refolding or, in the case of irreparable proteins, their removal by the protein degradation machinery of the cell. During their digenetic lifestyle, Leishmania parasites encounter and adapt to harsh environmental conditions, such as nutrient deficiency, hypoxia, oxidative stress, changing pH, and shifts in temperature; all these factors are potential triggers of cellular stress. We summarize here our current knowledge on the main types of molecular chaperones in Leishmania and their functions. Among them, heat shock proteins play important roles in adaptation and survival of this parasite against temperature changes associated with its passage from the poikilothermic insect vector to the warm-blooded vertebrate host. The study of structural features and the function of chaperones in Leishmania biology is providing opportunities (and challenges) for drug discovery and improving of current treatments against leishmaniasis. Jose M. Requena, Ana M. Montalvo, and Jorge Fraga Copyright © 2015 Jose M. Requena et al. All rights reserved. Immunology and Cell Biology of Parasitic Diseases 2014 Wed, 20 May 2015 07:00:21 +0000 Luis I. Terrazas, Abhay R. Satoskar, Miriam Rodriguez-Sosa, and Abraham Landa-Piedra Copyright © 2015 Luis I. Terrazas et al. All rights reserved. Iron and Parasites Wed, 20 May 2015 06:56:13 +0000 Rossana Arroyo, Theresa Ochoa, Jung-Hsiang Tai, and Mireya de la Garza Copyright © 2015 Rossana Arroyo et al. All rights reserved. Role of the Ubiquitin-Proteasome Systems in the Biology and Virulence of Protozoan Parasites Tue, 19 May 2015 12:37:40 +0000 In eukaryotic cells, proteasomes perform crucial roles in many cellular pathways by degrading proteins to enforce quality control and regulate many cellular processes such as cell cycle progression, signal transduction, cell death, immune responses, metabolism, protein-quality control, and development. The catalytic heart of these complexes, the 20S proteasome, is highly conserved in bacteria, yeast, and humans. However, until a few years ago, the role of proteasomes in parasite biology was completely unknown. Here, we summarize findings about the role of proteasomes in protozoan parasites biology and virulence. Several reports have confirmed the role of proteasomes in parasite biological processes such as cell differentiation, cell cycle, proliferation, and encystation. Proliferation and cell differentiation are key steps in host colonization. Considering the importance of proteasomes in both processes in many different parasites such as Trypanosoma, Leishmania, Toxoplasma, and Entamoeba, parasite proteasomes might serve as virulence factors. Several pieces of evidence strongly suggest that the ubiquitin-proteasome pathway is also a viable parasitic therapeutic target. Research in recent years has shown that the proteasome is a valid drug target for sleeping sickness and malaria. Then, proteasomes are a key organelle in parasite biology and virulence and appear to be an attractive new chemotherapeutic target. Christian Muñoz, Juan San Francisco, Bessy Gutiérrez, and Jorge González Copyright © 2015 Christian Muñoz et al. All rights reserved. Effect of Clinoptilolite and Sepiolite Nanoclays on Human and Parasitic Highly Phagocytic Cells Tue, 19 May 2015 09:30:04 +0000 Nanoclays have potential applications in biomedicine raising the need to evaluate their toxicity in in vitro models as a first approach to its biocompatibility. In this study, in vitro toxicity of clinoptilolite and sepiolite nanoclays (NC) was analyzed in highly phagocytic cultures of amoebas and human and mice macrophages. While amebic viability was significantly affected only by sepiolite NC at concentrations higher than 0.1 mg/mL, the effect on macrophage cultures was dependent on the origin of the cells. Macrophages derived from human peripheral blood monocytes were less affected in viability (25% decrease at 48 h), followed by the RAW 264.7 cell line (40%), and finally, macrophages derived from mice bone marrow monocytes (98%). Moreover, the cell line and mice macrophages die mainly by necrosis, whereas human macrophages exhibit increased apoptosis. Cytokine expression analysis in media of sepiolite NC treated cultures showed a proinflammatory profile (INFγ, IL-1α, IL-8, and IL-6), in contrast with clinoptilolite NC that induced lees cytokines with concomitant production of IL-10. The results show that sepiolite NC is more toxic to amoebas and macrophages than clinoptilolite NC, mostly in a time and dose-dependent manner. However, the effect of sepiolite NC was comparable with talc powder suggesting that both NC have low cytotoxicity in vitro. Yanis Toledano-Magaña, Leticia Flores-Santos, Georgina Montes de Oca, Alfonso González-Montiel, Juan-Pedro Laclette, and Julio-César Carrero Copyright © 2015 Yanis Toledano-Magaña et al. All rights reserved. Characterization of a Thioredoxin-1 Gene from Taenia solium and Its Encoding Product Tue, 19 May 2015 09:05:58 +0000 Taenia solium thioredoxin-1 gene (TsTrx-1) has a length of 771 bp with three exons and two introns. The core promoter gene presents two putative stress transcription factor binding sites, one putative TATA box, and a transcription start site (TSS). TsTrx-1 mRNA is expressed higher in larvae than in adult. This gene encodes a protein of 107 amino acids that presents the Trx active site (CGPC), the classical secondary structure of the thioredoxin fold, and the highest degree of identity with the Echinococcus granulosus Trx. A recombinant TsTrx-1 (rTsTrx-1) was produced in Escherichia coli with redox activity. Optimal activity for rTsTrx-1 was at pH 6.5 in the range of 15 to 25°C. The enzyme conserved activity for 3 h and lost it in 24 h at 37°C. rTsTrx-1 lost 50% activity after 1 h and lost activity completely in 24 h at temperatures higher than 55°C. Best storage temperature for rTsTrx-1 was at −70°C. It was inhibited by high concentrations of H2O2 and methylglyoxal (MG), but it was inhibited neither by NaCl nor by anti-rTsTrx-1 rabbit antibodies that strongly recognized a ~12 kDa band in extracts from several parasites. These TsTrx-1 properties open the opportunity to study its role in relationship T. solium-hosts. Lucía Jiménez, Oscar Rodríguez-Lima, Alicia Ochoa-Sánchez, and Abraham Landa Copyright © 2015 Lucía Jiménez et al. All rights reserved. Protection against Amoebic Liver Abscess in Hamster by Intramuscular Immunization with an Autographa californica Baculovirus Driving the Expression of the Gal-Lectin LC3 Fragment Tue, 19 May 2015 08:20:23 +0000 In a previous study, we demonstrated that oral immunization using Autographa californica baculovirus driving the expression of the Gal-lectin LC3 fragment (AcNPV-LC3) of Entamoeba histolytica conferred protection against ALA development in hamsters. In this study, we determined the ability of AcNPV-LC3 to protect against ALA by the intramuscular route as well as the liver immune response associated with protection. Results showed that 55% of hamsters IM immunized with AcNPV-LC3 showed sterile protection against ALA, whereas other 20% showed reduction in the size and extent of abscesses, resulting in some protection in 75% of animals compared to the sham control group. Levels of protection showed a linear correlation with the development and intensity of specific antiamoeba cellular and humoral responses, evaluated in serum and spleen of hamsters, respectively. Evaluation of the Th1/Th2 cytokine patterns expressed in the liver of hamsters showed that sterile protection was associated with the production of high levels of IFNγ and IL-4. These results suggest that the baculovirus system is equally efficient by the intramuscular as well as the oral routes for ALA protection and that the Gal-lectin LC3 fragment is a highly protective antigen against hepatic amoebiasis through the local induction of IFNγ and IL-4. Dulce María Meneses-Ruiz, Hugo Aguilar-Diaz, Raúl José Bobes, Alicia Sampieri, Luis Vaca, Juan Pedro Laclette, and Julio César Carrero Copyright © 2015 Dulce María Meneses-Ruiz et al. All rights reserved. Iron-Binding Protein Degradation by Cysteine Proteases of Naegleria fowleri Mon, 18 May 2015 14:39:52 +0000 Naegleria fowleri causes acute and fulminant primary amoebic meningoencephalitis. This microorganism invades its host by penetrating the olfactory mucosa and then traveling up the mesaxonal spaces and crossing the cribriform plate; finally, the trophozoites invade the olfactory bulbs. During its invasion, the protozoan obtains nutrients such as proteins, lipids, carbohydrates, and cationic ions (e.g., iron, calcium, and sodium) from the host. However, the mechanism by which these ions are obtained, particularly iron, is poorly understood. In the present study, we evaluated the ability of N. fowleri to degrade iron-binding proteins, including hololactoferrin, transferrin, ferritin, and hemoglobin. Zymography assays were performed for each substrate under physiological conditions (pH 7 at 37°C) employing conditioned medium (CM) and total crude extracts (TCEs) of N. fowleri. Different degradation patterns with CM were observed for hololactoferrin, transferrin, and hemoglobin; however, CM did not cause ferritin degradation. In contrast, the TCEs degraded only hololactoferrin and transferrin. Inhibition assays revealed that cysteine proteases were involved in this process. Based on these results, we suggest that CM and TCEs of N. fowleri degrade iron-binding proteins by employing cysteine proteases, which enables the parasite to obtain iron to survive while invading the central nervous system. Moisés Martínez-Castillo, Gerardo Ramírez-Rico, Jesús Serrano-Luna, and Mineko Shibayama Copyright © 2015 Moisés Martínez-Castillo et al. All rights reserved. Trichomonas vaginalis Cysteine Proteinases: Iron Response in Gene Expression and Proteolytic Activity Mon, 18 May 2015 12:30:14 +0000 We focus on the iron response of Trichomonas vaginalis to gene family products such as the cysteine proteinases (CPs) involved in virulence properties. In particular, we examined the effect of iron on the gene expression regulation and function of cathepsin L-like and asparaginyl endopeptidase-like CPs as virulence factors. We addressed some important aspects about CPs genomic organization and we offer possible explanations to the fact that only few members of this large gene family are expressed at the RNA and protein levels and the way to control their proteolytic activity. We also summarized all known iron regulations of CPs at transcriptional, posttranscriptional, and posttranslational levels along with new insights into the possible epigenetic and miRNA processes. Rossana Arroyo, Rosa Elena Cárdenas-Guerra, Elisa Elvira Figueroa-Angulo, Jonathan Puente-Rivera, Olga Zamudio-Prieto, and Jaime Ortega-López Copyright © 2015 Rossana Arroyo et al. All rights reserved. Clinicopathological and Immunological Changes in Indian Post Kala-Azar Dermal Leishmaniasis (PKDL) Cases in relation to Treatment: A Retrospective Study Mon, 18 May 2015 12:12:48 +0000 Post-kala-azar dermal leishmaniasis (PKDL) is an important factor in kala-azar transmission; hence its early detection and assessment of effective treatment is very important for disease control. In present study on 60 PKDL cases presented with macular, mixed papulonodular, or erythematous lesions, Leishmania parasites were demonstrated microscopically in 91% of papulonodular and 40% of macular lesions. Cellular infiltrates in skin biopsy imprint smears from lesions were mononuclear cells, 25–300/OIF (oil immersion field), predominantly histiocytes with vacuolation, many lymphocytes, some plasma cells, and Leishmania amastigotes 0–20/OIF. Cases with no demonstrable parasites were diagnosed on the basis of past history of VL, lesion’s distribution, cytopathological changes, and positive DAT (86.83%). Following antileishmanial treatment with SAG, papulonodular forms of PKDL lesions disappeared clinically but microscopically the mononuclear cells (20–200/OIF) persisted in the dermal lesions. Response observed in macular PKDL lesions was poor which persisted both clinically and cytopathologically. Follow-up of PKDL will assess the effectivity of treatment as either disappearance of lesions or any relapse. Studies on involvement of immunological factors, that is, certain cytokines (IL-10, TGF-β, etc.) and chemokines (macrophage inflammatory protein, MIP 1-, etc.) in PKDL, may provide insight for any role in the treatment response. Neena Verma, Sanjiv Bimal, Vidya Nand Rabi Das, Krishna Pandey, Dharmendra Singh, Chandra Shekhar Lal, Ashish Kumar Singh, Prabhat Kumar Sinha, and Pradeep Das Copyright © 2015 Neena Verma et al. All rights reserved. CK2 Secreted by Leishmania braziliensis Mediates Macrophage Association Invasion: A Comparative Study between Virulent and Avirulent Promastigotes Mon, 18 May 2015 12:11:49 +0000 CK2 is a protein kinase distributed in different compartments of Leishmania braziliensis: an externally oriented ecto-CK2, an intracellular CK2, and a secreted CK2. This latter form is constitutively secreted from the parasite (CsCK2), but such secretion may be highly enhanced by the association of specific molecules, including enzyme substrates, which lead to a higher enzymatic activity, called inductively secreted CK2 (IsCK2). Here, we examined the influence of secreted CK2 (sCK2) activity on the infectivity of a virulent L. braziliensis strain. The virulent strain presented 121-fold higher total CK2 activity than those found in an avirulent strain. The use of specific CK2 inhibitors (TBB, DRB, or heparin) inhibited virulent parasite growth, whereas no effect was observed in the avirulent parasites. When these inhibitors were added to the interaction assays between the virulent L. braziliensis strain and macrophages, association index was drastically inhibited. Polyamines enhanced sCK2 activity and increased the association index between parasites and macrophages. Finally, sCK2 and the supernatant of the virulent strain increased the association index between the avirulent strain and macrophages, which was inhibited by TBB. Thus, the kinase enzyme CK2 seems to be important to invasion mechanisms of L. braziliensis. Ana Madeira Brito Zylbersztejn, Carlos Gustavo Vieira de Morais, Ana Karina Castro Lima, Joyce Eliza de Oliveira Souza, Angela Hampshire Lopes, Sílvia Amaral Gonçalves Da-Silva, Mário Alberto Cardoso Silva-Neto, and Patrícia Maria Lourenço Dutra Copyright © 2015 Ana Madeira Brito Zylbersztejn et al. All rights reserved. Transfection of Platyhelminthes Mon, 18 May 2015 12:09:51 +0000 Flatworms are one of the most diverse groups within Lophotrochozoa with more than 20,000 known species, distributed worldwide in different ecosystems, from the free-living organisms in the seas and lakes to highly specialized parasites living in a variety of hosts, including humans. Several infections caused by flatworms are considered major neglected diseases affecting countries in the Americas, Asia, and Africa. For several decades, a particular interest on free-living flatworms was due to their ability to regenerate considerable portions of the body, implying the presence of germ cells that could be important for medicine. The relevance of reverse genetics for this group is clear; understanding the phenotypic characteristics of specific genes will shed light on developmental traits of free-living and parasite worms. The genetic manipulation of flatworms will allow learning more about the mechanisms for tissue regeneration, designing new and more effective anthelmintic drugs, and explaining the host-parasite molecular crosstalk so far partially inaccessible for experimentation. In this review, availability of transfection techniques is analyzed across flatworms, from the initial transient achievements to the stable manipulations now developed for free-living and parasite species. Bárbara Moguel, Raúl J. Bobes, Julio C. Carrero, and Juan P. Laclette Copyright © 2015 Bárbara Moguel et al. All rights reserved. Extraintestinal Helminth Infection Limits Pathology and Proinflammatory Cytokine Expression during DSS-Induced Ulcerative Colitis: A Role for Alternatively Activated Macrophages and Prostaglandins Mon, 18 May 2015 11:39:56 +0000 Chronic inflammation of the intestinal mucosa is characteristic of inflammatory bowel diseases such as ulcerative colitis and Crohn’s disease. Helminth parasites have developed immunomodulatory strategies that may impact the outcome of several inflammatory diseases. Therefore, we investigated whether Taenia crassiceps infection is able to decrease the inflammatory effects of dextran sulfate sodium- (DSS-) induced ulcerative colitis in BALB/c and C57BL/6 mice. Preinfection significantly reduced the manifestations of DSS-induced colitis, as weight loss and shortened colon length, and decreased the disease activity index independently of the genetic background of the mice. Taenia infection decreased systemic levels of proinflammatory cytokines while increasing levels of IL-4 and IL-10, and the inflammatory infiltrate into the colon was also markedly reduced. RT-PCR assays from colon showed that T. crassiceps-infected mice displayed increased expression of Arginase-1 but decreased expression of iNOS compared to DSS-treated uninfected mice. The percentages of T regulatory cells were not increased. The adoptive transfer of alternatively activated macrophages (AAMs) from infected mice into mice with DSS-induced colitis reduced the severity of colon inflammation. Administration of indomethacin abrogated the anticolitic effect of Taenia. Thus, T. crassiceps infection limits the pathology of ulcerative colitis by suppressing inflammatory responses mechanistically associated with AAMs and prostaglandins. Yadira Ledesma-Soto, Blanca E. Callejas, César A. Terrazas, Jose L. Reyes, Arlett Espinoza-Jiménez, Marisol I. González, Sonia León-Cabrera, Rosario Morales, Jonadab E. Olguín, Rafael Saavedra, Steve Oghumu, Abhay R. Satoskar, and Luis I. Terrazas Copyright © 2015 Yadira Ledesma-Soto et al. All rights reserved. Transferrin: Endocytosis and Cell Signaling in Parasitic Protozoa Mon, 18 May 2015 09:55:26 +0000 Iron is the fourth most abundant element on Earth and the most abundant metal in the human body. This element is crucial for life because almost all organisms need iron for several biological activities. This is the case with pathogenic organisms, which are at the vanguard in the battle with the human host for iron. The latest regulates Fe concentration through several iron-containing proteins, such as transferrin. The transferrin receptor transports iron to each cell that needs it and maintains it away from pathogens. Parasites have developed several strategies to obtain iron as the expression of specific transferrin receptors localized on plasma membrane, internalized through endocytosis. Signal transduction pathways related to the activation of the receptor have functional importance in proliferation. The study of transferrin receptors and other proteins with action in the signaling networks is important because these proteins could be used as therapeutic targets due to their specificity or to differences with the human counterpart. In this work, we describe proteins that participate in signal transduction processes, especially those that involve transferrin endocytosis, and we compare these processes with those found in T. brucei, T. cruzi, Leishmania spp., and E. histolytica parasites. Magda Reyes-López, Carolina Piña-Vázquez, and Jesús Serrano-Luna Copyright © 2015 Magda Reyes-López et al. All rights reserved. Strategies of Intracellular Pathogens for Obtaining Iron from the Environment Mon, 18 May 2015 09:51:52 +0000 Most microorganisms are destroyed by the host tissues through processes that usually involve phagocytosis and lysosomal disruption. However, some organisms, called intracellular pathogens, are capable of avoiding destruction by growing inside macrophages or other cells. During infection with intracellular pathogenic microorganisms, the element iron is required by both the host cell and the pathogen that inhabits the host cell. This minireview focuses on how intracellular pathogens use multiple strategies to obtain nutritional iron from the intracellular environment in order to use this element for replication. Additionally, the implications of these mechanisms for iron acquisition in the pathogen-host relationship are discussed. Nidia Leon-Sicairos, Ruth Reyes-Cortes, Alma M. Guadrón-Llanos, Jesús Madueña-Molina, Claudia Leon-Sicairos, and Adrian Canizalez-Román Copyright © 2015 Nidia Leon-Sicairos et al. All rights reserved. Binding and Endocytosis of Bovine Hololactoferrin by the Parasite Entamoeba histolytica Mon, 18 May 2015 09:42:07 +0000 Entamoeba histolytica is a human parasite that requires iron (Fe) for its metabolic function and virulence. Bovine lactoferrin (B-Lf) and its peptides can be found in the digestive tract after dairy products are ingested. The aim of this study was to compare virulent trophozoites recently isolated from hamster liver abscesses with nonvirulent trophozoites maintained for more than 30 years in cultures in vitro regarding their interaction with iron-charged B-Lf (B-holo-Lf). We performed growth kinetics analyses of trophozoites in B-holo-Lf and throughout several consecutive transfers. The virulent parasites showed higher growth and tolerance to iron than nonvirulent parasites. Both amoeba variants specifically bound B-holo-Lf with a similar Kd. However, averages of 9.45 × 105 and 6.65 × 106 binding sites/cell were found for B-holo-Lf in nonvirulent and virulent amoebae, respectively. Virulent amoebae bound more efficiently to human and bovine holo-Lf, human holo-transferrin, and human and bovine hemoglobin than nonvirulent amoebae. Virulent amoebae showed two types of B-holo-Lf binding proteins. Although both amoebae endocytosed this glycoprotein through clathrin-coated vesicles, the virulent amoebae also endocytosed B-holo-Lf through a cholesterol-dependent mechanism. Both amoeba variants secreted cysteine proteases cleaving B-holo-Lf. These data demonstrate that the B-Lf endocytosis is more efficient in virulent amoebae. Guillermo Ortíz-Estrada, Víctor Calderón-Salinas, Mineko Shibayama-Salas, Nidia León-Sicairos, and Mireya de la Garza Copyright © 2015 Guillermo Ortíz-Estrada et al. All rights reserved. Iron Homeostasis and Trypanosoma brucei Associated Immunopathogenicity Development: A Battle/Quest for Iron Mon, 18 May 2015 09:35:56 +0000 African trypanosomosis is a chronic debilitating disease affecting the health and economic well-being of developing countries. The immune response during African trypanosome infection consisting of a strong proinflammatory M1-type activation of the myeloid phagocyte system (MYPS) results in iron deprivation for these extracellular parasites. Yet, the persistence of M1-type MYPS activation causes the development of anemia (anemia of chronic disease, ACD) as a most prominent pathological parameter in the mammalian host, due to enhanced erythrophagocytosis and retention of iron within the MYPS thereby depriving iron for erythropoiesis. In this review we give an overview of how parasites acquire iron from the host and how iron modulation of the host MYPS affects trypanosomosis-associated anemia development. Finally, we also discuss different strategies at the level of both the host and the parasite that can/might be used to modulate iron availability during African trypanosome infections. Benoit Stijlemans, Alain Beschin, Stefan Magez, Jo A. Van Ginderachter, and Patrick De Baetselier Copyright © 2015 Benoit Stijlemans et al. All rights reserved. Corrigendum to “Somatostatin Negatively Regulates Parasite Burden and Granulomatous Responses in Cysticercosis” Mon, 18 May 2015 09:04:39 +0000 Mitra Khumbatta, Bahrom Firozgary, David John Tweardy, Joel Weinstock, Gohar Firozgary, Zal Bhatena, Tushar Bulsara, Ricardo Siller, and Prema Robinson Copyright © 2015 Mitra Khumbatta et al. All rights reserved. The Dialogue of the Host-Parasite Relationship: Leishmania spp. and Trypanosoma cruzi Infection Mon, 18 May 2015 08:39:31 +0000 The intracellular protozoa Leishmania spp. and Trypanosoma cruzi and the causative agents of Leishmaniasis and Chagas disease, respectively, belong to the Trypanosomatidae family. Together, these two neglected tropical diseases affect approximately 25 million people worldwide. Whether the host can control the infection or develops disease depends on the complex interaction between parasite and host. Parasite surface and secreted molecules are involved in triggering specific signaling pathways essential for parasite entry and intracellular survival. The recognition of the parasite antigens by host immune cells generates a specific immune response. Leishmania spp. and T. cruzi have a multifaceted repertoire of strategies to evade or subvert the immune system by interfering with a range of signal transduction pathways in host cells, which causes the inhibition of the protective response and contributes to their persistence in the host. The current therapeutic strategies in leishmaniasis and trypanosomiasis are very limited. Efficacy is variable, toxicity is high, and the emergence of resistance is increasingly common. In this review, we discuss the molecular basis of the host-parasite interaction of Leishmania and Trypanosoma cruzi infection and their mechanisms of subverting the immune response and how this knowledge can be used as a tool for the development of new drugs. Carlos Gustavo Vieira de Morais, Ana Karina Castro Lima, Rodrigo Terra, Rosiane Freire dos Santos, Silvia Amaral Gonçalves Da-Silva, and Patrícia Maria Lourenço Dutra Copyright © 2015 Carlos Gustavo Vieira de Morais et al. All rights reserved. Morphological Findings in Trophozoites during Amoebic Abscess Development in Misoprostol-Treated BALB/c Mice Mon, 18 May 2015 08:37:17 +0000 During amoebic liver abscess (ALA) formation in susceptible animals, immune response is regulated by prostaglandin E2 (PGE2) dependent mechanisms. The aim of this study was to analyze the effect of misoprostol (MPL), a PGE1 analogue, on ALA formation in BALB/c mice. Male mice from BALB/c strain were intrahepatically infected with trophozoites of E. histolytica strain HM1:IMSS and treated with 10−4 M of MPL daily until sacrifice at 2, 4, and 7 days postinfection (p.i.). ALA formation was evaluated at 2, 4, and 7 days postinfection; trophozoite morphology was analyzed using immunohistochemistry and image analysis. Results showed an increase in frequency of ALA formation in infected and MPL-treated mice only at 2 days p.i. (). A significant diminution in the size of trophozoites was detected in abscesses from mice independently of MPL treatment (from  µm at 2 days p.i. to  µm at 7 days p.i.) compared with trophozoites dimensions observed in susceptible hamsters ( µm) (). These results suggest that MPL treatment may modify the adequate control of inflammatory process to allow the persistence of trophozoites in the liver; however, natural resistance mechanisms cannot be discarded. Andrés Aceves-Cano, Rocío Gaytán-Ochoa, Ernesto Ramos-Martínez, Gilberto Erosa de la Vega, Carmen González-Horta, Patricia Talamás-Rohana, and Blanca Sánchez-Ramírez Copyright © 2015 Andrés Aceves-Cano et al. All rights reserved. Prevalence and Risk Factors of Intestinal Parasites in Cats from China Mon, 11 May 2015 14:17:24 +0000 The prevalence of intestinal parasites in cats from China was largely unknown prior to this study. The aim of the present study was to investigate the presence of intestinal parasites in cats from central China and also identify risk factors for parasitism. Fecal samples from 360 cats were examined using sugar flotation procedure and fecal smear test by microscope. Cats had mixed two or three kinds of parasites infections. Of the 360 cats feces, intestinal parasites positive feces were 149 (41.39%). 64 (17.78%) were infected with Toxocara cati, 61 (16.94%) with Isospora felis, 41 (11.39%) with Isospora rivolta, 33 (9.17%) with Paragonimus, 23 (6.39%) with hookworms, 11 (3.06%) with Toxoplasma-like oocysts, 10 (2.78%) with Trichuris, 4 (1.11%) with lungworm, 2 (0.56%) with Sarcocystis, and 1 (0.28%) with Trematode. The cats’ living outdoor was identified as risk factor by statistical analysis. These results provide relevant basic data for assessing the infection of intestinal parasites in cats from central region of China. In conclusion, there was high prevalence of intestinal parasites in cats from China. Yurong Yang and Hongde Liang Copyright © 2015 Yurong Yang and Hongde Liang. All rights reserved. Advances in Development of New Treatment for Leishmaniasis Mon, 11 May 2015 06:27:25 +0000 Leishmaniasis is a neglected infectious disease caused by several different species of protozoan parasites of the genus Leishmania. Current strategies to control this disease are mainly based on chemotherapy. Despite being available for the last 70 years, leishmanial chemotherapy has lack of efficiency, since its route of administration is difficult and it can cause serious side effects, which results in the emergence of resistant cases. The medical-scientific community is facing difficulties to overcome these problems with new suitable and efficient drugs, as well as the identification of new drug targets. The availability of the complete genome sequence of Leishmania has given the scientific community the possibility of large-scale analysis, which may lead to better understanding of parasite biology and consequent identification of novel drug targets. In this review we focus on how high-throughput analysis is helping us and other groups to identify novel targets for chemotherapeutic interventions. We further discuss recent data produced by our group regarding the use of the high-throughput techniques and how this helped us to identify and assess the potential of new identified targets. Juliana Perrone Bezerra de Menezes, Carlos Eduardo Sampaio Guedes, Antônio Luis de Oliveira Almeida Petersen, Deborah Bittencourt Mothé Fraga, and Patrícia Sampaio Tavares Veras Copyright © 2015 Juliana Perrone Bezerra de Menezes et al. All rights reserved. Activation of Sonic Hedgehog Leads to Survival Enhancement of Astrocytes via the GRP78-Dependent Pathway in Mice Infected with Angiostrongylus cantonensis Thu, 16 Apr 2015 06:22:26 +0000 Angiostrongylus cantonensis infection may cause elevation of ROS and antioxidants in the CSF of infected mice. Astrocytes may protect the surrounding neurons from oxidative stress-induced cell death by secreting Sonic hedgehog (Shh) via the PI3-K/AKT/Bcl-2 pathway. This study was conducted to determine the role of the Shh signaling pathway in A. cantonensis-infected BABL/c mice by coculturing astrocytes with living fifth-stage larvae or soluble antigens. The Shh pathway was activated with corresponding increases in the level of the Shh. Glial fibrillary acidic protein (GFAP) and Shh were increased in astrocyte cocultured with living fifth-stage larvae or soluble antigens. The survival of astrocytes pretreated with Shh was significantly elevated in cocultures with the antigens but reduced by its inhibitor cyclopamine. The expression of GRP78 and Bcl-2 was significantly higher in astrocytes pretreated with recombinant Shh. These findings suggest that the expression of Shh may inhibit cell death by activating Bcl-2 through a GRP78-dependent pathway. Kuang-Yao Chen, Chien-Ju Cheng, and Lian-Chen Wang Copyright © 2015 Kuang-Yao Chen et al. All rights reserved. A New Methodology for Evaluation of Nematode Viability Thu, 19 Mar 2015 13:05:26 +0000 Nematodes infections are responsible for debilitating conditions and economic losses in domestic animals as well as livestock and are considered an important public health problem due to the high prevalence in humans. The nematode resistance for drugs has been reported for livestock, highlighting the importance for development of new anthelmintic compounds. The aim of the current study was to apply and compare fluorimetric techniques using Sytox and propidium iodide for evaluating the viability of C. elegans larvae after treatment with anthelmintic drugs. These fluorescent markers were efficient to stain larvae treated with ivermectin and albendazole sulfoxide. We observed that densitometric values were proportional to the concentration of dead larvae stained with both markers. Furthermore, data on motility test presented an inverse correlation with fluorimetric data when ivermectin was used. Our results showed that lower concentrations of drugs were effective to interfere in the processes of cellular transport while higher drugs concentrations were necessary in order to result in any damage to cell integrity. The methodology described in this work might be useful for studies that aim to evaluate the viability of nematodes, particularly for testing of new anthelminthic compounds using an easy, economic, reproducible, and no time-consuming technique. Sebastião Rodrigo Ferreira, Tiago Antônio Oliveira Mendes, Lilian Lacerda Bueno, Jackson Victor de Araújo, Daniella Castanheira Bartholomeu, and Ricardo Toshio Fujiwara Copyright © 2015 Sebastião Rodrigo Ferreira et al. All rights reserved. Phlebotomine Sand Fly Fauna and Leishmania Infection in the Vicinity of the Serra do Cipó National Park, a Natural Brazilian Heritage Site Sun, 22 Feb 2015 11:03:31 +0000 In the New World, the leishmaniases are primarily transmitted to humans through the bites of Leishmania-infected Lutzomyia (Diptera: Psychodidae) phlebotomine sand flies. Any or both of two basic clinical forms of these diseases are endemic to several cities in Brazil—the American cutaneous leishmaniasis (ACL) and the American visceral leishmaniasis (AVL). The present study was conducted in the urban area of a small-sized Brazilian municipality (Jaboticatubas), in which three cases of AVL and nine of ACL have been reported in the last five years. Jaboticatubas is an important tourism hub, as it includes a major part of the Serra do Cipó National Park. Currently, no local data is available on the entomological fauna or circulating Leishmania. During the one-year period of this study, we captured 3,104 phlebotomine sand flies belonging to sixteen Lutzomyia species. In addition to identifying incriminated or suspected vectors of ACL with DNA of the etiological agent of AVL and vice versa, we also detected Leishmania DNA in unexpected Lutzomyia species. The expressive presence of vectors and natural Leishmania infection indicates favorable conditions for the spreading of leishmaniases in the vicinity of the Serra do Cipó National Park. Rosana Silva Lana, Érika Monteiro Michalsky, Consuelo Latorre Fortes-Dias, João Carlos França-Silva, Fabiana de Oliveira Lara-Silva, Ana Cristina Vianna Mariano da Rocha Lima, Daniel Moreira de Avelar, Juliana Cristina Dias Martins, and Edelberto Santos Dias Copyright © 2015 Rosana Silva Lana et al. All rights reserved. Prevalence of Protozoa Species in Drinking and Environmental Water Sources in Sudan Wed, 18 Feb 2015 09:20:34 +0000 Protozoa are eukaryotic cells distributed worldwide in nature and are receiving increasing attention as reservoirs and potential vectors for the transmission of pathogenic bacteria. In the environment, on the other hand, many genera of the protozoa are human and animal pathogens. Only limited information is available on these organisms in developing countries and so far no information on their presence is available from Sudan. It is necessary to establish a molecular identification of species of the protozoa from drinking and environmental water. 600 water samples were collected from five states (Gadarif, Khartoum, Kordofan, Juba, and Wad Madani) in Sudan and analysed by polymerase chain reaction (PCR) and sequencing. 57 out of 600 water samples were PCR positive for protozoa. 38 out of the 57 positive samples were identified by sequencing to contain 66 protozoa species including 19 (28.8%) amoebae, 17 (25.7%) Apicomplexa, 25 (37.9%) ciliates, and 5 (7.6%) flagellates. This study utilized molecular methods identified species belonging to all phyla of protozoa and presented a fast and accurate molecular detection and identification of pathogenic as well as free-living protozoa in water uncovering hazards facing public health. Salah Shanan, Hadi Abd, Magdi Bayoumi, Amir Saeed, and Gunnar Sandström Copyright © 2015 Salah Shanan et al. All rights reserved. Control of Avian Coccidiosis: Future and Present Natural Alternatives Tue, 17 Feb 2015 14:35:15 +0000 Numerous efforts to date have been implemented in the control of avian coccidiosis caused by the Eimeria parasite. Since the appearance of anticoccidial chemical compounds, the search for new alternatives continues. Today, no product is available to cope with the disease; however, the number of products commercially available is constantly increasing. In this review, we focus on natural products and their anticoccidial activity. This group comprises fatty acids, antioxidants, fungal and herbal extracts, and immune response modulators with proven anticoccidial activity, many of which exist as dietary supplements. Additionally, we offer an overview of the poultry industry and the economic cost of coccidiosis as well as the classical strategies used to control the disease. Rosa Estela Quiroz-Castañeda and Edgar Dantán-González Copyright © 2015 Rosa Estela Quiroz-Castañeda and Edgar Dantán-González. All rights reserved. Immunodiagnosis of Canine Visceral Leishmaniasis Using Mimotope Peptides Selected from Phage Displayed Combinatorial Libraries Thu, 29 Jan 2015 13:34:38 +0000 ELISA and RIFI are currently used for serodiagnosis of canine visceral leishmaniasis (CVL). The accuracy of these tests is controversial in endemic areas where canine infections by Trypanosoma cruzi may occur. We evaluated the usefulness of synthetic peptides that were selected through phage display technique in the serodiagnosis of CVL. Peptides were chosen based on their ability to bind to IgGs purified from infected dogs pooled sera. We selected three phage clones that reacted only with those IgGs. Peptides were synthesized, polymerized with glutaraldehyde, and used as antigens in ELISA assays. Each individual peptide or a mix of them was reactive with infected dogs serum. The assay was highly sensitive and specific when compared to soluble Leishmania antigen that showed cross-reactivity with anti-T. cruzi IgGs. Our results demonstrate that phage display technique is useful for selection of peptides that may represent valuable synthetic antigens for an improved serodiagnosis of CVL. Christina Monerat Toledo-Machado, Ricardo Andrez Machado de Avila, Christophe NGuyen, Claude Granier, Lilian Lacerda Bueno, Claudia Martins Carneiro, Daniel Menezes-Souza, Rubens Antonio Carneiro, Carlos Chávez-Olórtegui, and Ricardo Toshio Fujiwara Copyright © 2015 Christina Monerat Toledo-Machado et al. All rights reserved. Analysis of the Isomerase and Chaperone-Like Activities of an Amebic PDI (EhPDI) Wed, 28 Jan 2015 07:05:45 +0000 Protein disulfide isomerases (PDI) are eukaryotic oxidoreductases that catalyze the formation and rearrangement of disulfide bonds during folding of substrate proteins. Structurally, PDI enzymes share as a common feature the presence of at least one active thioredoxin-like domain. PDI enzymes are also involved in holding, refolding, and degradation of unfolded or misfolded proteins during stressful conditions. The EhPDI enzyme (a 38 kDa polypeptide with two active thioredoxin-like domains) has been used as a model to gain insights into protein folding and disulfide bond formation in E. histolytica. Here, we performed a functional complementation assay, using a ΔdsbC mutant of E. coli, to test whether EhPDI exhibits isomerase activity in vivo. Our preliminary results showed that EhPDI exhibits isomerase activity; however, further mutagenic analysis revealed significant differences in the functional role of each thioredoxin-like domain. Additional studies confirmed that EhPDI protects heat-labile enzymes against thermal inactivation, extending our knowledge about its chaperone-like activity. The characterization of EhPDI, as an oxidative folding catalyst with chaperone-like function, represents the initial step to dissect the molecular mechanisms involved in protein folding in E. histolytica. Rosa E. Mares, Alexis Z. Minchaca, Salvador Villagrana, Samuel G. Meléndez-López, and Marco A. Ramos Copyright © 2015 Rosa E. Mares et al. All rights reserved. In Vitro Metacyclogenesis of Leishmania (Viannia) braziliensis and Leishmania (Leishmania) amazonensis Clinical Field Isolates, as Evaluated by Morphology, Complement Resistance, and Infectivity to Human Macrophages Wed, 28 Jan 2015 06:49:24 +0000 This study was designed to assess in vitro metacyclogenesis of Leishmania (Viannia) braziliensis and Leishmania (Leishmania) amazonensis clinical field isolates obtained from patient lesions (L. braziliensis IMG3 and PPS6m; L. amazonensis MAB6). Metacyclogenesis was evaluated by different criteria, namely, promastigote size (morphometric analysis and flow cytometry), surface modifications (loss of lectin or monoclonal antibody (mAb) binding, complement resistance), and infectivity to human macrophages. Growth curves were similar for all parasites evaluated. The various features analyzed were expressed in a high percentage of promastigotes at 6th and 10th days of culture and a low percentage at the 2nd day. However, in most isolates, these features, considered as markers of metacyclogenesis, seemed to develop with different time courses, since the percentages of metacyclic forms detected with each technique were usually different. Parasites from 6th or 10th day and those negatively selected with lectin or mAb similarly infected human macrophages. From all isolates analyzed, L. amazonensis PH8 and MAB6 showed the highest and the lowest levels of susceptibility, respectively, to leishmanicidal activity of IFN-γ/LPS-activated macrophages. Our results showed that by using different techniques to evaluate different aspects of metacyclogenesis (morphological and biochemical modifications) different percentages of metacyclic promastigotes can be detected in each isolate culture. Ildefonso Alves da Silva Jr., Camila Imai Morato, Valéria Bernadete Leite Quixabeira, Ledice Inácia de Araújo Pereira, Miriam Leandro Dorta, Milton Adriano Pelli de Oliveira, Maria Fátima Horta, and Fátima Ribeiro-Dias Copyright © 2015 Ildefonso Alves da Silva Jr. et al. All rights reserved. Pathogenic Potential of Two Sibling Species, Anisakis simplex (s.s.) and Anisakis pegreffii (Nematoda: Anisakidae): In Vitro and In Vivo Studies Wed, 21 Jan 2015 08:29:18 +0000 The pathogenic potentials of two sibling nematodes Anisakis simplex sensu stricto (s.s.) and A. pegreffii were compared by in vitro and in vivo studies. Live third-stage larvae of each species were subjected to agar blocks made using PBS or RPMI-1640, overlaid with different supernatants (artificial gastric juice, PBS, and RPMI-1640), and their penetration ability was compared. Their tolerance of artificial gastric juice was also tested. Further, they were introduced into rats by gastric intubation, and the in vivo locations of them were investigated. A. pegreffii showed higher penetration ability than A. simplex (s.s.) in most of the experimental conditions, except for the RPMI-1640 agar block overlaid with artificial gastric juice. In an acid tolerance test, the mean survival times were 6.1 days for A. simplex (s.s.) and 4.2 days for A. pegreffii. In an animal experiment, A. simplex (s.s.) stayed for a shorter time in the stomachs of rats than A. pegreffii. Some A. pegreffii and A. simplex (s.s.) were embedded in the gastric mucosa or freely existed in the abdominal cavity. All of these results suggest that A. pegreffii has the pathogenic potential to cause anisakidosis in humans when ingested, as does A. simplex (s.s.). Chan-Hyeok Jeon and Jeong-Ho Kim Copyright © 2015 Chan-Hyeok Jeon and Jeong-Ho Kim. All rights reserved. The Macrophage Galactose-Type Lectin-1 (MGL1) Recognizes Taenia crassiceps Antigens, Triggers Intracellular Signaling, and Is Critical for Resistance to This Infection Thu, 15 Jan 2015 10:21:07 +0000 C-type lectins are multifunctional sugar-binding molecules expressed on dendritic cells (DCs) and macrophages that internalize antigens for processing and presentation. Macrophage galactose-type lectin 1 (MGL1) recognizes glycoconjugates expressing Lewis X structures which contain galactose residues, and it is selectively expressed on immature DCs and macrophages. Helminth parasites contain large amounts of glycosylated components, which play a role in the immune regulation induced by such infections. Macrophages from MGL1−/− mice showed less binding ability toward parasite antigens than their wild-type (WT) counterparts. Exposure of WT macrophages to T. crassiceps antigens triggered tyrosine phosphorylation signaling activity, which was diminished in MGL1−/− macrophages. Following T. crassiceps infection, MGL1−/− mice failed to produce significant levels of inflammatory cytokines early in the infection compared to WT mice. In contrast, MGL1−/− mice developed a Th2-dominant immune response that was associated with significantly higher parasite loads, whereas WT mice were resistant. Flow cytometry and RT-PCR analyses showed overexpression of the mannose receptors, IL-4Rα, PDL2, arginase-1, Ym1, and RELM-α on MGL1−/− macrophages. These studies indicate that MGL1 is involved in T. crassiceps recognition and subsequent innate immune activation and resistance. Daniel Montero-Barrera, Héctor Valderrama-Carvajal, César A. Terrazas, Saúl Rojas-Hernández, Yadira Ledesma-Soto, Laura Vera-Arias, Maricela Carrasco-Yépez, Lorena Gómez-García, Diana Martínez-Saucedo, Mireya Becerra-Díaz, and Luis I. Terrazas Copyright © 2015 Daniel Montero-Barrera et al. All rights reserved. Antioxidant and Anti-Inflammatory Activities of Pomegranate (Punica granatum) on Eimeria papillata-Induced Infection in Mice Thu, 15 Jan 2015 07:04:54 +0000 Coccidiosis is the most prevalent disease causing widespread economic loss, especially in poultry farms. Here, we investigated the effects of pomegranate peel extract (PPE) on the outcome of coccidiosis caused by Eimeria papillata in mice. The data showed that mice infected with E. papillata and treated with PPE revealed a significant decrease in the output of oocysts in their faeces by day 5 p.i. Infection also induced inflammation and injury of the jejunum. This was evidenced (i) as increases in reactive oxygen species, (ii), as increased neutrophils and decreased lymphocytes in blood (ii) as increased mRNA levels of inducible nitric oxide synthase (iNOS), Bcl-2 gene, and of the cytokines interferon gamma (IFN-γ), tumour necrosis factor-α (TNF-α), and interleukin-1β (IL-1β), and (iv) as downregulation of mucin gene MUC2 mRNA. All these infection-induced parameters were significantly altered during PPE treatment. In particular, PPE counteracted the E. papillata-induced loss of the total antioxidant capacity. Our data indicated that PPE treatment significantly attenuated inflammation and injury of the jejunum induced by E. papillata infections. Omar S. O. Amer, Mohamed A. Dkhil, Wafaa M. Hikal, and Saleh Al-Quraishy Copyright © 2015 Omar S. O. Amer et al. All rights reserved. Stress Management in Cyst-Forming Free-Living Protists: Programmed Cell Death and/or Encystment Mon, 12 Jan 2015 12:20:26 +0000 In the face of harsh conditions and given a choice, a cell may (i) undergo programmed cell death, (ii) transform into a cancer cell, or (iii) enclose itself into a cyst form. In metazoans, the available evidence suggests that cellular machinery exists only to execute or avoid programmed cell death, while the ability to form a cyst was either lost or never developed. For cyst-forming free-living protists, here we pose the question whether the ability to encyst was gained at the expense of the programmed cell death or both functions coexist to counter unfavorable environmental conditions with mutually exclusive phenotypes. Naveed Ahmed Khan, Junaid Iqbal, and Ruqaiyyah Siddiqui Copyright © 2015 Naveed Ahmed Khan et al. All rights reserved. Dendritic Cell Profile Induced by Schistosoma mansoni Antigen in Cutaneous Leishmaniasis Patients Wed, 17 Sep 2014 06:41:25 +0000 The inflammatory response in cutaneous leishmaniasis (CL), although responsible for controlling the infection, is associated with the pathogenesis of disease. Conversely, the immune response induced by S. mansoni antigens is able to prevent immune-mediated diseases. The aim of this study was to evaluate the potential of the S. mansoni Sm29 antigen to change the profile of monocyte-derived dendritic cells (MoDCs) from subjects with cutaneous leishmaniasis (CL) in vitro. Monocytes derived from the peripheral blood mononuclear cells of twelve patients were cultured with GM-CSF and IL-4 for differentiation into dendritic cells and then stimulated with soluble Leishmania antigen (SLA) in the presence or absence of Sm29 antigen. The expression of surface molecules associated with maturation and activation (HLA-DR, CD40, CD83, CD80, and CD86), inflammation (IL-12, TNF), and downregulation (IL-10, IL-10R) was evaluated using flow cytometry. We observed that the frequencies of HLA-DR, CD83, CD80, and CD86 as well as of IL-10 and IL-10R on MoDCs were higher in cultures stimulated with Sm29, compared to the unstimulated cell cultures. Our results indicate that the Sm29 antigen is able to activate regulatory MoDCs in patients with cutaneous leishmaniasis. It might be useful to control the inflammatory process associated with this disease. Diego Mota Lopes, Jamille Souza Fernandes, Thiago Marconi de Souza Cardoso, Aline Michele Barbosa Bafica, Sérgio Costa Oliveira, Edgar M. Carvalho, Maria Ilma Araujo, and Luciana Santos Cardoso Copyright © 2014 Diego Mota Lopes et al. All rights reserved. Differentially Expressed Genes of Virulent and Nonvirulent Entamoeba histolytica Strains Identified by Suppression Subtractive Hybridization Wed, 17 Sep 2014 00:00:00 +0000 Entamoeba histolytica is a parasite which presents capacity to degrade tissues and therefore has a pathogenic behavior. As this behavior is not shown by all strains, there have been several studies investigating molecular basis of the cytotoxicity process. Using the suppression subtractive hybridization (SSH) technique, differential gene expressions of two E. histolytica strains, one virulent (EGG) and one nonvirulent (452), have been analyzed with the purpose of isolating genes which may be involved with amoebic virulence. Nine cDNA fragments presenting high homology with E. histolytica previously sequenced genes were subtracted. Of these, four genes were confirmed by RT-PCR. Two coding for hypothetical proteins, one for a cysteine-rich protein, expressed only in the virulent strain, EGG and another one, coding for grainin 2 protein, exclusive from 452 strain. This study provided new insight into the proteins differences in the virulent and nonvirulent E. histolytica strains. We believe that further studies with these proteins may prove association of them with tissue damage, providing new perceptions to improve treatment or diagnosis of the invasive disease. Michelle A. R. Freitas, Ângela C. Alvarenga, Helen C. Fernandes, Frederico F. Gil, Maria N. Melo, Jorge L. Pesquero, and Maria A. Gomes Copyright © 2014 Michelle A. R. Freitas et al. All rights reserved. The Associations of Leishmania major and Leishmania tropica Aspects by Focusing Their Morphological and Molecular Features on Clinical Appearances in Khuzestan Province, Iran Tue, 16 Sep 2014 00:00:00 +0000 Cutaneous leishmaniasis has various phenotypic aspects consisting of polymorphic amastigotes with different genetic ranges. Samples were collected from suspected patients of Khuzestan province. Prepared smears were stained, scaled, and measured using ocular micrometer. The Cyt b, ITS-rDNA, and microsatellite genes of Leishmania were amplified and Leishmania species were identified by molecular analyses. Of 150 examined suspected patients, 102 were identified to Leishmania species (90 L. major, nine L. tropica, and three unidentified). The amastigotes of 90 L. major had regular and different irregular shapes within three clinical lesions with no and/or low genetic diversity. Three haplotypes of Cyt b of L. major were found but no variation was observed using ITS-rDNA gene. Interesting findings were that all nine L. tropica had regular amastigote shapes with more genetic variations, also a patient which had coinfection of L. major, L. tropica, and Crithidia. At least two L. major and L. tropica were identified in suspected patients of the regions. Different irregular amastigotes’ shapes of L. major can be explained by various reservoir hosts and vectors. In contrast, more molecular variations in L. tropica could be justified by genetic characters. Unidentified Leishmania could be mixed pathogens or nonpathogens with mammals’ Leishmania or Crithidia. Adel Spotin, Soheila Rouhani, and Parviz Parvizi Copyright © 2014 Adel Spotin et al. All rights reserved. Helminth Parasites Alter Protection against Plasmodium Infection Mon, 08 Sep 2014 00:00:00 +0000 More than one-third of the world’s population is infected with one or more helminthic parasites. Helminth infections are prevalent throughout tropical and subtropical regions where malaria pathogens are transmitted. Malaria is the most widespread and deadliest parasitic disease. The severity of the disease is strongly related to parasite density and the host’s immune responses. Furthermore, coinfections between both parasites occur frequently. However, little is known regarding how concomitant infection with helminths and Plasmodium affects the host’s immune response. Helminthic infections are frequently massive, chronic, and strong inductors of a Th2-type response. This implies that infection by such parasites could alter the host’s susceptibility to subsequent infections by Plasmodium. There are a number of reports on the interactions between helminths and Plasmodium; in some, the burden of Plasmodium parasites increased, but others reported a reduction in the parasite. This review focuses on explaining many of these discrepancies regarding helminth-Plasmodium coinfections in terms of the effects that helminths have on the immune system. In particular, it focuses on helminth-induced immunosuppression and the effects of cytokines controlling polarization toward the Th1 or Th2 arms of the immune response. Víctor H. Salazar-Castañon, Martha Legorreta-Herrera, and Miriam Rodriguez-Sosa Copyright © 2014 Víctor H. Salazar-Castañon et al. All rights reserved. Protective Effect of a Prime-Boost Strategy with the Ts87 Vaccine against Trichinella spiralis Infection in Mice Thu, 28 Aug 2014 15:20:04 +0000 Trichinellosis is a widespread zoonosis primarily caused by Trichinella spiralis. Mucosal immunity is crucial for preventing Trichinella spiralis infection. In our previous study, a DNA vaccine with the Trichinella antigen Ts87 delivered by an attenuated Salmonella typhimurium elicited partial protection against Trichinella spiralis infection in mice. In the current study, to elicit a more robust immune response and develop a potent vaccination strategy against trichinellosis, a heterologous prime-boost vaccination regimen for Ts87 was used in mice and the protective efficacy was evaluated compared to the homologous DNA prime-boost or protein prime-boost immunization alone. The results revealed that the DNA-prime/protein-boost vaccination with Ts87 induced higher levels of both humoral and cellular immune responses. The challenge results showed that mice with the DNA-prime/protein-boost vaccination displayed higher muscle larval reduction than those immunized with DNA prime-boost or protein prime-boost. The results demonstrated that mice vaccinated with Ts87 in a DNA-prime/protein-boost strategy effectively elicited a local IgA response and mixed Th1/Th2 immune response that might be responsible for improved protection against Trichinella spiralis infection. Yuan Gu, Bin Zhan, Yaping Yang, Xiaodi Yang, Xi Zhao, Lei Wang, Jing Yang, Kuo Bi, Yunyun Wang, and Xinping Zhu Copyright © 2014 Yuan Gu et al. All rights reserved. Diethylstilbestrol Exposure in Neonatal Mice Induces Changes in the Adulthood in the Immune Response to Taenia crassiceps without Modifications of Parasite Loads Thu, 28 Aug 2014 06:31:45 +0000 Industrial growth has increased the exposition to endocrine disruptor compounds (EDC’s), which are exogenous agents with agonist or antagonist action of endogenous steroid hormones that may affect the course of parasite infections. We wanted to determine if the exposure to diethylstilbestrol (DES), an estrogen agonist, to both male and female mice affected the immune response and their susceptibility to T. crassiceps cysticercosis. In all infected groups, females showed higher parasite loads than males, and neonatal DES administration did not modify this pattern. In the spleen, noninfected mice showed sex-related differences in the percentage of the CD8+ subpopulation, but DES decreased the percentage of CD3+, CD19+, and CD8+ subpopulations in infected mice. In the mesenteric lymphatic node (MNL), DES showed a dimorphic effect in the percentage of CD19+ cells. Regarding estrogen receptor alpha (ER-α) expression, DES treatment induced a reduction in the expression of this receptor in both noninfected female and male mice in the spleen, which was decreased only in males in CD3+ and CD8+ lymphocytes in MNL cell subpopulations. Our study is the first one to demonstrate that DES neonatal treatment in male and female mice affects the immune cell percentage, without effect on the susceptibility to T. crassiceps cysticercosis. Karen E. Nava-Castro, Jorge Morales-Montor, Alejandra Ortega-Hernando, and Ignacio Camacho-Arroyo Copyright © 2014 Karen E. Nava-Castro et al. All rights reserved. Dynamic of the Cellular Immune Response at the Dermal Site of Leishmania (L.) amazonensis and Leishmania (V.) braziliensis Infection in Sapajus apella Primate Thu, 28 Aug 2014 06:26:38 +0000 The purpose of this study was to characterize the immunopathological response in the skin of S. apella infected with Leishmania (L.) amazonensis and L. (V.) braziliensis parasites, the main causative agents of localized cutaneous leishmaniasis in South America. In infected animals, amastigote forms of L. (L.) amazonensis could be detected till 120 days postinfection (PI), while, in L. (V.) braziliensis infection, parasites could be detected until 180 days PI in the skin sections. CD20+ cells were detected throughout the experimental time in both groups as well as in CD3+ cells, which appeared to be activated because high densities of inducible nitric oxide synthase (iNOS+) cells were detected at 60 and 90 days PI in both studied groups. After 60 and 120 days PI, decrease in iNOS+ cells was observed in L. (L.) amazonensis and L. (V.) braziliensis, respectively, which was associated with parasite clearance. Increase in lysozyme+ cells was observed during the experimental infections, which also can be associated with parasite killing. Márcia Dalastra Laurenti, Luiz Felipe Domingues Passero, Thaise Yumie Tomokane, Fernanda de Camargo Francesquini, Mussya Cisotto Rocha, Claudia Maria de Castro Gomes, Carlos Eduardo Pereira Corbett, and Fernando Tobias Silveira Copyright © 2014 Márcia Dalastra Laurenti et al. All rights reserved. Gonadal Steroids Negatively Modulate Oxidative Stress in CBA/Ca Female Mice Infected with P. berghei ANKA Wed, 27 Aug 2014 08:41:09 +0000 We decreased the level of gonadal steroids in female and male mice by gonadectomy. We infected these mice with P. berghei ANKA and observed the subsequent impact on the oxidative stress response. Intact females developed lower levels of parasitaemia and lost weight faster than intact males. Gonadectomised female mice displayed increased levels of parasitaemia, increased body mass, and increased anaemia compared with their male counterparts. In addition, gonadectomised females exhibited lower specific catalase, superoxide dismutase, and glutathione peroxidase activities in their blood and spleen tissues compared with gonadectomised males. To further study the oxidative stress response in P. berghei ANKA-infected gonadectomised mice, nitric oxide levels were assessed in the blood and spleen, and MDA levels were assessed in the spleen. Intact, sham-operated, and gonadectomised female mice exhibited higher levels of nitric oxide in the blood and spleen compared with male mice. MDA levels were higher in all of the female groups. Finally, gonadectomy significantly increased the oxidative stress levels in females but not in males. These data suggest that differential oxidative stress is influenced by oestrogens that may contribute to sexual dimorphism in malaria. Néstor Aarón Mosqueda-Romo, Ana Laura Rodríguez-Morales, Fidel Orlando Buendía-González, Margarita Aguilar-Sánchez, Jorge Morales-Montor, and Martha Legorreta-Herrera Copyright © 2014 Néstor Aarón Mosqueda-Romo et al. All rights reserved. High Molecular Weight Proteins of Trypanosoma cruzi Reduce Cross-Reaction with Leishmania spp. in Serological Diagnosis Tests Sun, 20 Jul 2014 00:00:00 +0000 Chagas disease is caused by the parasite Trypanosoma cruzi. Because of its distribution throughout Latin America, sometimes it can overlap with other parasitic diseases, such as leishmaniasis, caused by Leishmania spp. This might represent a problem when performing serological diagnosis, because both parasites share antigens, resulting in cross-reactions. In the present work we evaluated Mexican sera samples: 83.8% of chagasic patients recognized at least one antigen of high molecular weight (>95 kDa) when evaluated by Western blot. Proteins of 130 kDa and 160 kDa are predominantly being recognized by asymptomatic chagasic patients. When the proteins were extracted using Triton X-100 detergent, a larger number of specific T. cruzi proteins were obtained. This protein fraction can be used to increase specificity to 100% in Western blot assays without losing sensitivity of the test. High molecular weight proteins of T. cruzi include glycoproteins with a great amount of αMan (α-mannose), αGlc (α-glucose), GlcNAc (N-acetylglucosamine), and αGal (α-galactose) content and these structures play an essential role in antigens recognition by antibodies present in patients’ sera. Alejandra Yunuen Cervantes-Landín, Ignacio Martínez, Muslim Schabib, and Bertha Espinoza Copyright © 2014 Alejandra Yunuen Cervantes-Landín et al. All rights reserved. The Effect of Babesia divergens Infection on the Spleen of Mongolian Gerbils Thu, 17 Jul 2014 08:38:01 +0000 Babesiosis is caused by intraerythrocytic protozoan parasites transmitted by ticks and affects a wide range of domestic and wild animals and occasionally humans. The current study aimed to investigate the effect of B. divergens infected erythrocytes on spleen histopathology, cell cycle alteration, and the presence of oxidative stress. Mongolian gerbils were challenged with 5 × 106  Babesia divergens infected erythrocytes. Parasitemia reached approximately 77% at day 5 postinfection. Infection also induced injury of the spleen. This was evidenced with (i) increases in cellular damage of the spleen, (ii) decrease in antioxidant capacity as indicated by decreased glutathione, catalase, and superoxide dismutase levels, (iii) increased production of malondialdehyde and nitric oxide derived products (nitrite/nitrate), and (iv) increased lactic acid dehydrogenase activity and protein carbonyl content in the spleen. Infection interfered with normal cell cycle of the spleen cells at G0/G1, S, and G2/M phases. On the basis of the above results it can be hypothesized that B. divergens infected erythrocytes could alter the spleen histopathology and cause cell cycle alteration and induce oxidative stress in splenic tissue. Mohamed A. Dkhil, Saleh Al-Quraishy, and Mohamed S. Al-Khalifa Copyright © 2014 Mohamed A. Dkhil et al. All rights reserved. Somatostatin Negatively Regulates Parasite Burden and Granulomatous Responses in Cysticercosis Wed, 09 Jul 2014 00:00:00 +0000 Cysticercosis is an infection of tissues with the larval cysts of the cestode, Taenia  solium. While live parasites elicit little or no inflammation, dying parasites initiate a granulomatous reaction presenting as painful muscle nodules or seizures when cysts are located in the brain. We previously showed in the T. crassiceps murine model of cysticercosis that substance P (SP), a neuropeptide, was detected in early granulomas and was responsible for promoting granuloma formation, while somatostatin (SOM), another neuropeptide and immunomodulatory hormone, was detected in late granulomas; SOM’s contribution to granuloma formation was not examined. In the current studies, we used somatostatin knockout (SOM−/−) mice to examine the hypothesis that SOM downmodulates granulomatous inflammation in cysticercosis, thereby promoting parasite growth. Our results demonstrated that parasite burden was reduced 5.9-fold in SOM−/− mice compared to WT mice (). This reduction in parasite burden in SOM−/− mice was accompanied by a 95% increase in size of their granulomas (), which contained a 1.5-fold increase in levels of IFN-γ and a 26-fold decrease in levels of IL-1β ( for both) compared to granulomas from WT mice. Thus, SOM regulates both parasite burden and granulomatous inflammation perhaps through modulating granuloma production of IFN-γ and IL-1β. Mitra Khumbatta, Bahrom Firozgary, David John Tweardy, Joel Weinstock, Gohar Firozgary, Zal Bhatena, Tushar Bulsara, Ricardo Siller, and Prema Robinson Copyright © 2014 Mitra Khumbatta et al. All rights reserved. The Trend in Distribution of Q223R Mutation of Leptin Receptor Gene in Amoebic Liver Abscess Patients from North India: A Prospective Study Wed, 09 Jul 2014 00:00:00 +0000 Host genetic susceptibility is an important risk factor in infectious diseases. We explored the distribution of Q223R mutation in leptin receptor gene of amoebic liver abscess (ALA) patients of North India. A total of 55 ALA samples along with 102 controls were subjected to PCR-RFLP analysis. The frequency of allele “G” (coding for arginine) was in general high in Indian population irrespective of the disease. Our results of Fisher exact test shows that heterozygous mutant (QQ versus QR, ) and homozygous mutant (QQ versus RR, ) were significantly associated with amoebic liver abscess when compared with homozygous wild (QQ). Anil Kumar Verma, Vineet Ahuja, and Jaishree Paul Copyright © 2014 Anil Kumar Verma et al. All rights reserved. Cofactor Independent Phosphoglycerate Mutase of Brugia malayi Induces a Mixed Th1/Th2 Type Immune Response and Inhibits Larval Development in the Host Tue, 01 Jul 2014 08:24:41 +0000 Lymphatic filariasis is a major debilitating disease, endemic in 72 countries putting more than 1.39 billion people at risk and 120 million are already infected. Despite the significant progress in chemotherapeutic advancements, there is still need for other measures like development of an effective vaccine or discovery of novel drug targets. In this study, structural and immunological characterization of independent phosphoglycerate mutase of filarial parasite Brugia malayi was carried out. Protein was found to be expressed in all major parasite life stages and as an excretory secretory product of adult parasites. Bm-iPGM also reacted to all the categories of human bancroftian patient’s sera including endemic normals. In vivo immunological behaviour of protein was determined in immunized BALB/c mice followed by prophylactic analysis in BALB/c mice and Mastomys coucha. Immunization with Bm-iPGM led to generation of a mixed Th1/Th2 type immune response offering 58.2% protection against larval challenge in BALB/c and 65–68% protection in M. coucha. In vitro studies confirmed participation of anti-Bm-iPGM antibodies in killing of B. malayi infective larvae and microfilariae through ADCC mechanism. The present findings reveal potential immunoprotective nature of Bm-iPGM advocating its worth as an antifilarial vaccine candidate. Prashant K. Singh, Susheela Kushwaha, Ajay K. Rana, and Shailja Misra-Bhattacharya Copyright © 2014 Prashant K. Singh et al. All rights reserved. Trichinella spiralis Excretory-Secretory Products Protect against Polymicrobial Sepsis by Suppressing MyD88 via Mannose Receptor Thu, 26 Jun 2014 10:59:47 +0000 Trichinella spiralis (T. spiralis) or its excretory-secretory products (TsES) protect hosts from autoimmune diseases, which depend on inducing host T helper (Th) 2 immune response and inhibiting inflammatory factors. Sepsis is a systemic inflammatory response syndrome (SIRS) evoked by infection. Little is known about the effects of helminths or their excretory-secretory products on sepsis. Here, we investigated the effects of TsES in a mice model of polymicrobial sepsis. TsES improved survival, reduced organ injury, and enhanced bacterial clearance in septic mice. To investigate the molecular mechanism, macrophages from septic patients or the control group were incubated with TsES. TsES reduced sepsis-inducing inflammatory cytokines mediated by Toll-like receptors (TLR) in vitro by suppressing TLR adaptor-transducer myeloid differentiation factor 88 (MyD88) and nuclear factor- (NF-)-κB. Furthermore, TsES upregulated mannose receptor (MR) expression during sepsis. MR blocking attenuated the effects of TsES on MyD88 and NF-κB expression. In vivo, MR RNAi reduced the survival rate of septic mice treated with TsES, suggesting that TsES-mediated protection against polymicrobial sepsis is dependent on MR. Thus, TsES administration might be a potential therapeutic strategy for treating sepsis. Linlin Du, Lihua Liu, Yang Yu, Hui Shan, and Leiqing Li Copyright © 2014 Linlin Du et al. All rights reserved. Effect of Various Acaricides on Hatchability of Eggs of Rhipicephalus (Boophilus) microplus Thu, 26 Jun 2014 10:52:58 +0000 The effect of commonly used acaricides (amitraz, cypermethrin, deltamethrin, fenvalerate, and flumethrin) on the eggs of Rhipicephalus (Boophilus) microplus collected from Ludhiana, Punjab, was evaluated by egg hatch assay (EHA). The regression graph of probit hatchability and per cent inhibition of hatching (IH%) of eggs was plotted against log values of concentration of various acaricides. All concentrations of flumethrin and amitraz caused complete inhibition of hatching, whereas a hatchability of , and % was only recorded at the highest concentration of cypermethrin, deltamethrin, and fenvalerate, respectively. An increase in the concentration of the acaricide showed a significant effect on the IH% of eggs for cypermethrin and deltamethrin but was nonsignificant for fenvalerate. The slope of the regression curve of IH% was utilized for the calculation of the dose of various acaricides causing inhibition of hatching for 95% eggs (LC95) and the discriminating dose (DD). Results indicated that maximum DD was recorded for fenvalerate (2.136%), followed by cypermethrin (0.214%) and deltamethrin (0.118%). The results of the current study will be helpful in formulating effective control strategies against ticks. M. Haque, Jyoti, N. K. Singh, and S. S. Rath Copyright © 2014 M. Haque et al. All rights reserved. Canine Babesiosis in Northwestern India: Molecular Detection and Assessment of Risk Factors Thu, 12 Jun 2014 11:05:09 +0000 In the current study, a total of 214 blood samples from dogs in and around Ludhiana, Punjab (India), suspected for canine babesiosis were examined with conventional and molecular assays. Examination of Giemsa-stained peripheral thin blood smears revealed an overall prevalence of 7.47% (16/214) for canine babesiosis encompassing 0.93% (2/214) of large Babesia and 6.54% (14/214) of Babesia gibsoni. However, molecular diagnosis revealed 15.42% (33/214) samples positive for B. gibsoni infection as evident by the presence of 671 bp amplicon. The results of multivariate analysis showed that the prevalence of B. gibsoni was associated with various risk factors, namely, age (; OR: 0.398; CI 95%: 0.080–1.799), sex (; OR: 0.849; CI 95%: 0.403–1.791), breed of host (; OR: 3.345; CI 95%: 1.045–10.710), and season (; OR: 2.143; CI 95%: 0.788–5.830). The prevalence of B. gibsoni was higher in summer as compared to winter season and in younger dogs, while breed and sex of the host were not significantly associated with the occurrence of the disease. Amritpal Singh, Harkirat Singh, N. K. Singh, N. D. Singh, and S. S. Rath Copyright © 2014 Amritpal Singh et al. All rights reserved. Differential Expression and Immunolocalization of Antioxidant Enzymes in Entamoeba histolytica Isolates during Metronidazole Stress Thu, 12 Jun 2014 10:42:22 +0000 Entamoeba histolytica infections are endemic in the Indian subcontinent. Five to eight percent of urban population residing under poor sanitary conditions suffers from Entamoeba infections. Metronidazole is the most widely prescribed drug used for amoebiasis. In order to understand the impact of metronidazole stress on the parasite, we evaluated the expression of two antioxidant enzymes, peroxiredoxin and FeSOD, in Entamoeba histolytica isolates during metronidazole stress. The results reveal that, under metronidazole stress, the mRNA expression levels of these enzymes did not undergo any significant change. Interestingly, immunolocalization studies with antibodies targeting peroxiredoxin indicate differential localization of the protein in the cell during metronidazole stress. In normal conditions, all the Entamoeba isolates exhibit presence of peroxiredoxin in the nucleus as well as in the membrane; however with metronidazole stress the protein localized mostly to the membrane. The change in the localization pattern was more pronounced when the cells were subjected to short term metronidazole stress compared to cells adapted to metronidazole. The protein localization to the cell membrane could be the stress response mechanism in these isolates. Colocalization pattern of peroxiredoxin with CaBp1, a cytosolic protein, revealed that the membrane and nuclear localization was specific to peroxiredoxin during metronidazole stress. Lakshmi Rani Iyer, Nishant Singh, Anil Kumar Verma, and Jaishree Paul Copyright © 2014 Lakshmi Rani Iyer et al. All rights reserved. Erythrophagocytosis in Entamoeba histolytica and Entamoeba dispar: A Comparative Study Thu, 05 Jun 2014 07:32:50 +0000 Entamoeba histolytica is the causative agent of human intestinal and liver amebiasis. The extraordinary phagocytic activity of E. histolytica trophozoites has been accepted as one of the virulence mechanisms responsible for their invasive capacity. The recognition of the noninvasive Entamoeba dispar as a different species has raised the question as to whether the lack of pathogenic potential of this ameba correlates with a limited phagocytic capacity. We have therefore compared the process of erythrophagocytosis in both species by means of light and video microscopy, hemoglobin measurement, and the estimation of reactive oxygen species (ROS). In the present study, we confirmed that E. dispar has lower erythrophagocytic capacity. We also observed by video microscopy a new event of erythrocyte opsonization-like in both species, being more characteristic in E. histolytica. Moreover, E. dispar showed a lower capacity to produce ROS compared with the invasive species and also showed a large population of amoebae that did not engulf any erythrocyte over time. Our results demonstrate that E. histolytica has a higher phagocytic capacity than E. dispar, including a higher rate of production of ROS in the course of ingesting red blood cells. Daniel Talamás-Lara, Bibiana Chávez-Munguía, Arturo González-Robles, Patricia Talamás-Rohana, Lizbeth Salazar-Villatoro, Ángel Durán-Díaz, and Adolfo Martínez-Palomo Copyright © 2014 Daniel Talamás-Lara et al. All rights reserved. Zoonotic Neglected Tropical Diseases: New Approaches to Combat Old Enemies Thu, 05 Jun 2014 06:39:45 +0000 Fabio Ribeiro Braga, Pedro Mendoza de Gives, Adolfo Paz Silva, Filippe Elias de Freitas Soares, and Jackson Victor de Araújo Copyright © 2014 Fabio Ribeiro Braga et al. All rights reserved. Evaluation of Antibody Response to Various Developmental Stage Specific Somatic Antigens of Paramphistomum epiclitum in Goats Thu, 05 Jun 2014 00:00:00 +0000 Electrophoretic analysis of various developmental stage specific somatic antigens of Paramphistomum epiclitum (Digenea: Paramphistomidae), namely, metacercariae (McAg), immature intestinal flukes (ImIAg), immature ruminal flukes (ImRAg), and adult flukes (AAg), was done by native polyacrylamide gel electrophoresis. Result revealed presence of 3 (range 15.2–40.3 kDa), 13 (9.3–121.2 kDa), 14 (9.3–169.3 kDa), and 15 (8.0–169.3 kDa) polypeptides in McAg, ImIAg, ImRAg, and AAg, respectively. With an aim to identify a suitable immunodiagnostic antigen for early diagnosis of amphistomosis, the IgG antibody response to various developmental stage antigens in goats experimentally infected with metacercariae of P. epiclitum was evaluated by ELISA. The highest OD values were recorded with ImIAg which ranged between 0.23 and 0.55 with a significant increase from the 2nd week till 8th week of infection with a peak at 6th week. The analysis of statistical significance using a one-way analysis of variance with multiple pair wise comparisons revealed that IgG response was significantly higher with all antigens () except McAg () with a maximum mean difference of 0.1838 in comparison to control with ImIAg, thus, indicating that ImIAg which could be further exploited for its potential is a candidate for immunodiagnostic antigen for early diagnosis of amphistomosis. Jyoti, A. Prasad, and Nirbhay Kumar Singh Copyright © 2014 Jyoti et al. All rights reserved. Relationship between Leishmania IFAT Titer and Clinicopathological Manifestations (Clinical Score) in Dogs Tue, 03 Jun 2014 11:53:58 +0000 During canine leishmaniasis (CanL) due to Leishmania infantum, high levels of antibodies production are associated with the presence of various clinical signs, because of the deposition of soluble immune complexes in organs and tissues. The immunofluorescence antibody test (IFAT) is one of the most commonly used techniques for detection of anti-Leishmania antibodies. The purpose of this study was to assess whether there is a correlation between clinical signs and IFAT titers in dogs naturally infected with Leishmania. A retrospective study was performed on medical records of 49 dogs diagnosed with CanL. Information extracted from the medical records of each dog with CanL was clinical score, IFAT titer, serum total protein (TP), gamma globulin (IgG) and creatinine concentration, and protein creatinine ratio in urine sample (UP/UC) at each follow-up examination. Results show that dogs with highest IFAT titers recorded had higher mean clinical scores indicating a positive relationship () between anti-Leishmania antibodies (IgG) and clinical manifestations, which becomes more evident in severe clinical forms of canine leishmaniasis. Higher TP and IgG serum concentrations were recorded in dogs with higher clinical scores. Significant association was observed between UP/UC and the IFAT titer (). Daniela Proverbio, Eva Spada, Giada Bagnagatti de Giorgi, Roberta Perego, and Emanuela Valena Copyright © 2014 Daniela Proverbio et al. All rights reserved. Exacerbated Skeletal Muscle Inflammation and Calcification in the Acute Phase of Infection by Mexican Trypanosoma cruzi DTUI Strain Mon, 02 Jun 2014 11:36:27 +0000 A murine model was used to study the histopathological aspects and cytokine expression levels in skeletal muscle provoked by the infection with Mexican TcI strains. BALB/c mice were inoculated with the virulent Querétaro strain and the nonvirulent Ninoa strain. Parasite numbers were counted in blood and skeletal muscle at different times post-infection, and real time-PCR expression levels of the cytokines IL-12, IL-4, IL-10, IFN-γ, and TNF-α were evaluated. In the acute phase of infection, a high parasitic load, both in blood and skeletal muscle, was detected. The histopathological analyses showed an exacerbated inflammation and granulomatous-like infiltrate with the Querétaro strain. Interestingly, extensive calcification areas were observed in the skeletal muscle surrounded by inflammatory infiltrates. TNF-α and IL-10 expression exhibited a significant increase at the peak of infection. In summary, Querétaro strain, a Mexican TcI strain, is virulent enough to induce high inflammation and calcification in skeletal muscle of the hind limbs, which could be related to high expression levels of TNF-α. Andrea Vizcaíno-Castillo, Andrea Jiménez-Marín, and Bertha Espinoza Copyright © 2014 Andrea Vizcaíno-Castillo et al. All rights reserved. Crosstalk among Taenia crassiceps (ORF Strain) Cysts Regulates Their Rates of Budding by Ways of Soluble and Contact Signals Exchanged between Them Wed, 28 May 2014 10:45:43 +0000 Herein we report that in vitro experiments with different initial parasite densities (1, 5, and 10 cysts per mL of culture medium) show that cysts at densities of 10 and 5 grow faster than those at 1, and that they release into the culture medium factors which increase the budding rates of the slower lower-density ones. Close contact among the incubated cysts also favors budding, thus suggesting the participation of surface sensors of parasite crowding. Thus, contact signals, together with the release of soluble growth factors, could endow cysts with the capacity to sense and regulate their numbers inside their habitat in relation to their population density. Esquivel-Velázquez Marcela, Hernández Ricardo, Larralde Carlos, and Ostoa-Saloma Pedro Copyright © 2014 Esquivel-Velázquez Marcela et al. All rights reserved. Activity of Tea Tree (Melaleuca alternifolia) Essential Oil against L3 Larvae of Anisakis simplex Sun, 25 May 2014 11:19:52 +0000 Nematicidal activity of Melaleuca alternifolia essential oil, commonly known as tea tree oil (TTO), was assayed in vitro against L3 larvae of Anisakis simplex. The results showed a mortality of 100% for concentrations between 7 and 10 μL/mL after 48 h of incubation, obtaining an LD50 value of 4.53 μL/mL after 24 hours and 4.27 μL/mL after 48 hours. Concentration-dependent inhibition of acetylcholinesterase was observed for tea tree essential oil showing inhibition values of 100% at 100 μL/mL. This fact suggests that TTO may act as an AChE inhibitor. Terpinen-4-ol was discarded as main larvicide compound as it did not show larvicidal or anticholinesterase activity. The data obtained suggest that the essential oil of Melaleuca alternifolia may have a great therapeutic potential for the treatment of human anisakiasis. Carlota Gómez-Rincón, Elisa Langa, Paula Murillo, Marta Sofía Valero, César Berzosa, and Víctor López Copyright © 2014 Carlota Gómez-Rincón et al. All rights reserved. Presence of Leishmania and Brucella Species in the Golden Jackal Canis aureus in Serbia Thu, 22 May 2014 12:14:07 +0000 The golden jackal Canis aureus occurs in south-eastern Europe, Asia, the Middle East, the Caucasus, and Africa. In Serbia, jackals neared extinction; however, during the last 30 years, the species started to spread quickly and to increase in number. Few studies in the past have revealed their potential role as carriers of zoonotic diseases. Animal samples were collected over a three-year period (01/2010–02/2013) from 12 sites all over Serbia. Of the tissue samples collected, spleen was chosen as the tissue to proceed; all samples were tested for Leishmania species and Brucella species by real-time PCR. Of the 216 samples collected, 15 (6.9%) were positive for Leishmania species, while four (1.9%) were positive for B. canis. The potential epidemiologic role of the golden jackal in carrying and dispersing zoonotic diseases in Serbia should be taken under consideration when applying surveillance monitoring schemes. Duško Ćirović, Dimosthenis Chochlakis, Snežana Tomanović, Ratko Sukara, Aleksandra Penezić, Yannis Tselentis, and Anna Psaroulaki Copyright © 2014 Duško Ćirović et al. All rights reserved. Modified PEHPS Medium as an Alternative for the In Vitro Culture of Giardia lamblia Wed, 21 May 2014 12:49:15 +0000 Commercial culture media present interlot variations in biological activity. We have previously designed a homemade and economic culture medium, PEHPS medium, for the axenic cultivation of Entamoeba histolytica and Trichomonas vaginalis. Trophozoites of amoebae and trichomonads grow well in this medium. Furthermore, the medium is stable for several months when stored frozen or refrigerated. The objective of this work was to modify PEHPS medium to support the in vitro growth of Giardia lamblia. Inocula of 5 × 103 trophozoites/mL of G. lamblia were incubated at 36.5°C in modified PEHPS or TYI-S-33 medium. Then, the growths of the three Giardia strains in both media were compared. The logarithmic growth phase lasted 72 h; the mean yield of the strains ranged from 10.06 to 11.43 × 105Giardia trophozoites/mL, and the range of duplication time in the three strains was from 5.67 to 6.06 in modified PEHPS medium. These growth characteristics were not significantly different from those obtained with TYI-S-33 medium. We conclude that modified PEHPS medium might be used for the axenic cultivation of G. lamblia. Javier Vargas-Villarreal, Benito D. Mata-Cárdenas, Magda E. Hernández-García, Jesús N. Garza-González, Laura H. De La Garza-Salinas, and Francisco González-Salazar Copyright © 2014 Javier Vargas-Villarreal et al. All rights reserved. Canine Filarial Infections in a Human Brugia malayi Endemic Area of India Tue, 20 May 2014 09:44:22 +0000 A very high prevalence of microfilaremia of 42.68 per cent out of 164 canine blood samples examined was observed in Cherthala (of Alappuzha district of Kerala state), a known human Brugia malayi endemic area of south India. The species of canine microfilariae were identified as Dirofilaria repens, Brugia malayi, and Acanthocheilonema reconditum. D. repens was the most commonly detected species followed by B. pahangi. D. immitis was not detected in any of the samples examined. Based on molecular techniques, microfilariae with histochemical staining pattern of “local staining at anal pore and diffuse staining at central body” was identified as D. repens in addition to those showing acid phosphatase activity only at the anal pore. Even though B. malayi like acid phosphatase activity was observed in few dogs examined, they were identified as genetically closer to B. pahangi. Hence, the possibility of dogs acting as reservoirs of human B. malayi in this area was ruled out. Reghu Ravindran, Sincy Varghese, Suresh N. Nair, Vimalkumar M. Balan, Bindu Lakshmanan, Riyas M. Ashruf, Swaroop S. Kumar, Ajith Kumar K. Gopalan, Archana S. Nair, Aparna Malayil, Leena Chandrasekhar, Sanis Juliet, Devada Kopparambil, Rajendran Ramachandran, Regu Kunjupillai, and Showkath Ali M. Kakada Copyright © 2014 Reghu Ravindran et al. All rights reserved. Antileishmanial, Toxicity, and Phytochemical Evaluation of Medicinal Plants Collected from Pakistan Mon, 19 May 2014 12:43:58 +0000 Leishmaniasis is an important parasitic problem and is in focus for development of new drugs all over the world. Objective of the present study was to evaluate phytochemical, toxicity, and antileishmanial potential of Jurinea dolomiaea, Asparagus gracilis, Sida cordata, and Stellaria media collected from different areas of Pakistan. Dry powder of plants was extracted with crude methanol and fractionated with n-hexane, chloroform, ethyl acetate, n-butanol, and water solvents in escalating polarity order. Qualitative phytochemical analysis of different class of compounds, that is, alkaloids, saponins, terpenoids, anthraquinones, cardiac glycosides, coumarins, phlobatannins, flavonoids, phenolics, and tannins, was tested. Its appearance was observed varying with polarity of solvent used for fractionation. Antileishmanial activity was performed against Leishmania tropica KWH23 promastigote. Potent antileishmanial activity was observed for J. dolomiaea methanol extract ( μg/mL) in comparison to other plant extracts. However, J. dolomiaea “ethyl acetate fraction” was more active ( μg/mL) against Leishmania tropica KWH23 among all plant fractions as well as standard Glucantime drug ( μg/mL). All the plants extract and its derived fraction exhibited toxicity in safety range () in brine shrimp toxicity evaluation assay. Naseer Ali Shah, Muhammad Rashid Khan, and Akhtar Nadhman Copyright © 2014 Naseer Ali Shah et al. All rights reserved. Chromatographic Fingerprint Analysis and Effects of the Medicinal Plant Species Mitracarpus frigidus on Adult Schistosoma mansoni Worms Thu, 08 May 2014 16:13:51 +0000 The aims of this work were to evaluate the in vitro and in vivo schistosomicidal properties of the methanolic extract of the aerial parts of Mitracarpus frigidus (MFM) and to determine its HPLC profile. For the in vitro experiment, four pairs of adult worms, obtained from infected mice, were exposed to different concentrations of MFM (100 to 400 μg/mL) for 24 and 48 h and analyzed under an inverted microscope. For the in vivo experiment, mice were inoculated with cercariae and, 20 days after infection, MFM (100 and 300 mg/kg) was administered orally for the following 25 days. Mice were euthanized after 60 days. MFM showed in vitro schistosomicidal activity, exhibiting the opening of the gynaecophoral canal of some male schistosomes, the presence of contorted muscles, vesicles, and the darkening of the paired worms skin. In vivo experiments showed that MFM treatments significantly reduced total worm count, as praziquantel, showing a decrease in liver and spleen weight. Also, a significant reduction in granuloma density was observed. MFM treatment did not cause alterations in the liver function of either infected or noninfected mice. The HPLC chromatogram profile showed the presence of kaempferol-O-rutinoside, rutin, kaempferol, psychorubrin, and ursolic acid. Rodrigo Luiz Fabri, Danielle Maria de Oliveira Aragão, Jônatas Rodrigues Florêncio, Nícolas de Castro Campos Pinto, Ana Carolina Alves Mattos, Paulo Marcos Zech Coelho, Maria Christina Marques Nogueira Castañon, Eveline Gomes Vasconcelos, Priscila de Faria Pinto, and Elita Scio Copyright © 2014 Rodrigo Luiz Fabri et al. All rights reserved. A Few Good Reasons Why Species-Area Relationships Do Not Work for Parasites Thu, 08 May 2014 12:36:28 +0000 Several studies failed to find strong relationships between the biological and ecological features of a host and the number of parasite species it harbours. In particular, host body size and geographical range are generally only weak predictors of parasite species richness, especially when host phylogeny and sampling effort are taken into account. These results, however, have been recently challenged by a meta-analytic study that suggested a prominent role of host body size and range extent in determining parasite species richness (species-area relationships). Here we argue that, in general, results from meta-analyses should not discourage researchers from investigating the reasons for the lack of clear patterns, thus proposing a few tentative explanations to the fact that species-area relationships are infrequent or at least difficult to be detected in most host-parasite systems. The peculiar structure of host-parasite networks, the enemy release hypothesis, the possible discrepancy between host and parasite ranges, and the evolutionary tendency of parasites towards specialization may explain why the observed patterns often do not fit those predicted by species-area relationships. Giovanni Strona and Simone Fattorini Copyright © 2014 Giovanni Strona and Simone Fattorini. All rights reserved. Frequency of Piroplasms Babesia microti and Cytauxzoon felis in Stray Cats from Northern Italy Sun, 04 May 2014 11:40:20 +0000 Emerging diseases caused by piroplasms pose a health risk for man and other animals, and domestic cats have been proposed as potential reservoirs for some piroplasm infections. The aim of this study was to identify the frequency of the piroplasms Babesia microti and Cytauxzoon felis in stray cats from northern Italy and to identify possible risk factors associated with these infections. Blood samples from 260 stray cats enrolled in a trap-neuter-release (TNR) program in northern Italy were examined with conventional PCR for the presence of Babesia microti and Cytauxzoon felis DNA. No sample (0.0%) tested positive for C. felis, whilst B. microti DNA was detected in two samples (0.8%). Both infected cats were in good clinical condition and recovered well from the neutering surgery. One of these two cats had a triple coinfection with Babesia microti, Candidatus Mycoplasma haemominutum, and Anaplasma phagocytophilum. Evidence presented in this study indicates that the blood borne protozoans Babesia microti and Cytauxzoon felis are not widely distributed in stray cat populations in Milan, northern Italy, and that the significance of cats as a reservoir host for B. microti in this area is limited. Eva Spada, Daniela Proverbio, Paola Galluzzo, Roberta Perego, Giada Bagnagatti De Giorgi, Nora Roggero, and Santo Caracappa Copyright © 2014 Eva Spada et al. All rights reserved. The Zoonotic Risk of Ancylostoma ceylanicum Isolated from Stray Dogs and Cats in Guangzhou, South China Sun, 04 May 2014 07:51:43 +0000 Canine and feline hookworm infection is endemic in many countries with zoonotic transmission representing a potentially significant public health concern. However, there is limited data available on the zoonotic transmission of canine and feline hookworms in China. This study was conducted to evaluate the zoonotic risk of Ancylostoma ceylanicum isolated from stray dogs and cats in Guangzhou, south China. Primer pairs CAF/CAR were designed to amplify complete ITS sequences of obtained A. ceylanicum. The results were compared with fourteen ITS reference sequences of human-derived A. ceylanicum registered in GenBank, and phylogenetic trees were established by using NJ and ML methods. The sequence similarity of three dog-derived and five cat-derived A. ceylanicum with fourteen human-derived A. ceylanicum were 96.8%~100% and 97.8%~100%, respectively. Phylogenetic analysis placed A. ceylanicum isolated from dogs and cats in the same group with A. ceylanicum human isolates. Due to the ability of A. ceylanicum to cause a patent infection in humans, the zoonotic risk arising from dog and cat reservoirs to communities in this region should be determined. Yuanjia Liu, Guochao Zheng, Muhamd Alsarakibi, Xinheng Zhang, Wei Hu, Liqin Lin, Liping Tan, Qin Luo, Pengyun Lu, and Guoqing Li Copyright © 2014 Yuanjia Liu et al. All rights reserved. Intestinal Parasites of Owned Dogs and Cats from Metropolitan and Micropolitan Areas: Prevalence, Zoonotic Risks, and Pet Owner Awareness in Northern Italy Mon, 28 Apr 2014 13:18:06 +0000 Intestinal parasites of dogs and cats are cosmopolitan pathogens with zoonotic potential for humans. Our investigation considered their diffusion in dogs and cats from northern Italy areas, specifically the metropolitan area of Milan and two micropolitan areas of neighboring provinces. It included the study of the level of awareness in pet owners of the zoonotic potential from these parasites. A total of 409 fresh fecal samples were collected from household dogs and cats for copromicroscopic analysis and detection of Giardia duodenalis coproantigens. The assemblages of Giardia were also identified. A questionnaire about intestinal parasites biology and zoonotic potential was submitted to 185 pet owners. The overall prevalence of intestinal parasites resulted higher in cats (47.37%−60.42%) and dogs (57.41%−43.02%) from micropolitan areas than that from the metropolis of Milan (dogs: %; cats: %). The zoonotic parasites infecting pets under investigation were T. canis and T. cati, T. vulpis, Ancylostomatidae, and G. duodenalis assemblage A. Only 49.19% of pet owners showed to be aware of the risks for human health from canine and feline intestinal parasites. Parasitological results in pets and awareness determination in their owners clearly highlight how the role of veterinarians is important in indicating correct and widespread behaviors to reduce risks of infection for pets and humans in urban areas. Sergio Aurelio Zanzani, Alessia Libera Gazzonis, Paola Scarpa, Federica Berrilli, and Maria Teresa Manfredi Copyright © 2014 Sergio Aurelio Zanzani et al. All rights reserved. A Retrospective Study of Ectoparasitosis in Patients Referred to Imam Reza Hospital of Mashhad, Iran Tue, 22 Apr 2014 09:40:40 +0000 This cross-sectional study was performed on all patients suspected to be suffering from ectoparasites who were referred to the parasitology laboratory of Imam Reza Hospital of Mashhad during 15 years (April 1995 to April 2010). All patients’ data were collected from the questionnaires and then analyzed statistically. From 1814 suspected patients to be suffering from ectoparasites, 375 patients had scabies and, 99 suffer from pediculosis. The mean age of patients was . The most common age of scabies was 10–19 (27/7%) and pediculosis 0–9(9/6%) ( value = 0.00). The highest incidence of pediculosis was in women (3.6%) and scabies in men (13.7%) ( value = 0.00). Pediculosis is more common in children (9/9%) and scabies in workers (32%)( value = 0.00). Scabies and pediculosis were more prevalent in patients from Razavi Khorasan Province with 18.7% and 5%, respectively ( value = 0.08). Fariba Berenji, Narges Marvi-Moghadam, and Parisa Naghibozakerin Meibodi Copyright © 2014 Fariba Berenji et al. All rights reserved. Putative Bronchopulmonary Flagellated Protozoa in Immunosuppressed Patients Thu, 03 Apr 2014 13:16:41 +0000 Flagellated protozoa that cause bronchopulmonary symptoms in humans are commonly neglected. These protozoal forms which were presumed to be “flagellated protozoa” have been previously identified in immunosuppressed patients in a number of studies, but have not been certainly classified so far. Since no human cases of bronchopulmonary flagellated protozoa were reported from Turkey, we aimed to investigate these putative protozoa in immunosuppressed patients who are particularly at risk of infectious diseases. Bronchoalveolar lavage fluid samples of 110 immunosuppressed adult patients who were admitted to the Department of Chest Diseases, Hafsa Sultan Hospital of Celal Bayar University, Manisa, Turkey, were examined in terms of parasites by light microscopy. Flagellated protozoal forms were detected in nine (8.2%) of 110 cases. Metronidazole (500 mg b.i.d. for 30 days) was given to all positive cases and a second bronchoscopy was performed at the end of the treatment, which revealed no parasites. In conclusion, immunosuppressed patients with bronchopulmonary symptoms should attentively be examined with regard to flagellated protozoa which can easily be misidentified as epithelial cells. Ali Ahmet Kilimcioglu, Yavuz Havlucu, Nogay Girginkardesler, Pınar Çelik, Kor Yereli, and Ahmet Özbilgin Copyright © 2014 Ali Ahmet Kilimcioglu et al. All rights reserved. Potential Biomarkers and Their Applications for Rapid and Reliable Detection of Malaria Wed, 02 Apr 2014 07:16:33 +0000 Malaria has been responsible for the highest mortality in most malaria endemic countries. Even after decades of malaria control campaigns, it still persists as a disease of high mortality due to improper diagnosis and rapidly evolving drug resistant malarial parasites. For efficient and economical malaria management, WHO recommends that all malaria suspected patients should receive proper diagnosis before administering drugs. It is thus imperative to develop fast, economical, and accurate techniques for diagnosis of malaria. In this regard an in-depth knowledge on malaria biomarkers is important to identify an appropriate biorecognition element and utilize it prudently to develop a reliable detection technique for diagnosis of the disease. Among the various biomarkers, plasmodial lactate dehydrogenase and histidine-rich protein II (HRP II) have received increasing attention for developing rapid and reliable detection techniques for malaria. The widely used rapid detection tests (RDTs) for malaria succumb to many drawbacks which promotes exploration of more efficient economical detection techniques. This paper provides an overview on the current status of malaria biomarkers, along with their potential utilization for developing different malaria diagnostic techniques and advanced biosensors. Priyamvada Jain, Babina Chakma, Sanjukta Patra, and Pranab Goswami Copyright © 2014 Priyamvada Jain et al. All rights reserved. Individual Subject Meta-Analysis of Parameters for Giardia duodenalis Shedding in Animal Experimental Models Tue, 01 Apr 2014 07:53:36 +0000 Giardia duodenalis is a zoonotic protozoan parasite with public health importance worldwide. While articles about animal model infectivity have been published for G. duodenalis, the studies have used diverse protocols and parameters to evaluate the infectivity of this protozoan parasite. Hence, the objectives of this study were to (1) conduct a meta-analysis of published literature for cyst shedding and diarrhea outcomes in animal models and (2) develop recommendations to help standardize experimental dose response studies. Results showed that, for the outcome of cyst shedding in faeces, the covariates of infective stage (cyst versus trophozoite), Giardia dose, and the interactions between doses and infective stage, as well as dose and species of experimental host, were all significant ( value ≤ 0.05). This study suggests inoculation of the experimental host with cysts rather than trophozoites and administration of higher doses of Giardia will most likely result in cyst shedding. Based on the results of this meta-analysis, the infective stage (cyst versus trophozoite), parasite dose, and the interactions between dose and infective stage, as well as dose and species of experimental host, should be considered when designing experimental dose response studies that will assist in the study of zoonotic neglected tropical diseases globally. A. D. Adell, W. A. Miller, D. J. Harvey, E. Van Wormer, S. Wuertz, and P. A. Conrad Copyright © 2014 A. D. Adell et al. All rights reserved. Trend Analysis of Visceral Leishmaniasis at Addis Zemen Health Center, Northwest Ethiopia Tue, 25 Mar 2014 13:26:27 +0000 Background. Visceral leishmaniasis (VL) is a systemic disease caused by the Leishmania donovani complex. It is one of the fatal diseases if left untreated. In Ethiopia, there are many VL endemic foci. The aim of this study was to determine the trends of VL in the study area. Methodology. A retrospective study was conducted at Addis Zemen health center from September 2005 to August 2011. Data were collected from laboratory registration book and entered and analyzed by using SPSS version 20 software and value of ≤0.05 was considered statistically significant. Result. A total of 7161 VL suspected cases were reported in the study area. The overall prevalence of VL was 2801 (39.1%). Of the 2801 VL positive cases, the highest annual prevalence, 988 (46.8%), was reported in 2005 but the trend gradually decreases. Majority of the VL confirmed cases were in the age groups of 5–14 years and males were more affected. Conclusion. The prevalence of VL in the study area was high in early 2005 but, gradually, the trend has been decreased and it becomes one of VL endemic foci in Ethiopia. Yitayih Wondimeneh, Yegnasew Takele, Asmamaw Atnafu, Getachew Ferede, and Dagnachew Muluye Copyright © 2014 Yitayih Wondimeneh et al. All rights reserved. Comparison between Conventional and Real-Time PCR Assays for Diagnosis of Visceral Leishmaniasis Thu, 06 Feb 2014 09:19:33 +0000 The diagnosis of visceral leishmaniasis (VL) is a challenging issue and several studies worldwide have evaluated the different tools to reach a diagnostic solution. The polymerase chain reaction (PCR) has proven to be effective in detecting the genome of Leishmania species in different biological samples. In this study, we compared the conventional PCR and real-time PCR using the Sybr Green system and their application in molecular diagnosis of visceral leishmaniasis in peripheral blood as a biological sample. The genus-specific conserved region of kinetoplast DNA (kDNA) was the target of amplification. We studied 30 samples from patients with suspect of visceral leishmaniasis who were treated by the Medical Clinic of Santa Casa de Belo Horizonte Hospital, Brazil. Among the samples studied, 19 had a confirmed diagnosis for VL by serology and/or by clinical findings. Among these 19 samples, 63% () presented positive results for serology and 79% () positive results in both PCR methodologies. This fact suggests that the PCR technique can assist in the diagnosis of visceral leishmaniasis in patients who do not have detectable antibodies by serology but can present the genome of the parasite circulating in whole blood. Also, it was possible to observe that there was conformity between the results of the techniques of cPCR and qPCR using the Sybr Green system in 100% of samples analyzed. These data suggest that both PCR techniques were equally effective for detection of the genome of the parasite in the patient’s blood. Mariana R. Pereira, Fabiana Rocha-Silva, Cidiane Graciele-Melo, Camila R. Lafuente, Telcia Magalhães, and Rachel B. Caligiorne Copyright © 2014 Mariana R. Pereira et al. All rights reserved. Detection of Toxoplasma gondii by PCR and Mouse Bioassay in Rodents of Ahvaz District, Southwestern Iran Wed, 29 Jan 2014 06:52:04 +0000 Toxoplasma gondii is obligate coccidian zoonotic parasite. Felidae family is definitive and wide ranges of warm-blooded vertebrates are intermediate hosts for the parasite. Rodents are measured as an important source of T. gondii infection for the definitive host. Thus, this study aimed to investigate Toxoplasm infection in rodents of Ahvaz district, southwest of Iran. A total of 100 rodents (73 Rattus norvegicus, 21 Rattus rattus, and 6 Mus musculus) were collected and studied by GRA6PCR and mouse bioassay. The finding indicated that 6 out of 100 (6%) and 2 out of 100 (2%) samples were positive by PCR and mouse bioassay, respectively. The results show notable chronic infection in the rodent and potential transmission of the infection among animal and men in the region. Accordingly, this study recommended investigating of the T. gondii infection in definitive and other intermediate hosts in other points of Khuzestan province, Southwest, Iran. J. Saki and S. Khademvatan Copyright © 2014 J. Saki and S. Khademvatan. All rights reserved. Diversity and Seasonal Impact of Acanthamoeba Species in a Subtropical Rivershed Mon, 30 Dec 2013 15:25:29 +0000 This study evaluated the presence of Acanthamoeba species in the Puzih River watershed, which features typical subtropical monsoon climate and is located just above the Tropic of Cancer in Taiwan. The relationship between the seasonal and geographical distributions of Acanthamoeba species in this rivershed was also investigated. Acanthamoeba species were detected in water samples using the amoebal enrichment culture method and confirmed by PCR. A total of 136 water samples were included in this study, 16 (11.7%) of which contained Acanthamoeba species. Samples with the highest percentage of Acanthamoeba (32.4%) were obtained during the summer season, mainly from upstream areas. The identified species in the four seasons included Acanthamoeba palestinensis (T2), Acanthamoeba sp. IS2/T4 (T4), Acanthamoeba lenticulata (T5), Acanthamoeba hatchetti (T11), Acanthamoeba healyi (T12), and Acanthamoeba jacobsi (T15). The most frequently identified Acanthamoeba genotype was T4 (68.7%). Acanthamoeba genotype T4 is responsible for Acanthamoeba keratitis and should be considered for associated human health risk potential in the rivershed. Po-Min Kao, Ming-Yuan Chou, Chi-Wei Tao, Wen-Chien Huang, Bing-Mu Hsu, Shu-Min Shen, Cheng-Wei Fan, and Yi-Chou Chiu Copyright © 2013 Po-Min Kao et al. All rights reserved. Analysis of Structures, Functions, and Epitopes of Cysteine Protease from Spirometra erinaceieuropaei Spargana Thu, 12 Dec 2013 11:34:31 +0000 Spirometra erinaceieuropaei cysteine protease (SeCP) in sparganum ES proteins recognized by early infection sera was identified by MALDI-TOF/TOF-MS. The aim of this study was to predict the structures and functions of SeCP protein by using the full length cDNA sequence of SeCP gene with online sites and software programs. The SeCP gene sequence was of 1 053 bp length with a 1011 bp biggest ORF encoding 336-amino acid protein with a complete cathepsin propeptide inhibitor domain and a peptidase C1A conserved domain. The predicted molecular weight and isoelectric point of SeCP were 37.87 kDa and 6.47, respectively. The SeCP has a signal peptide site and no transmembrane domain, located outside the membrane. The secondary structure of SeCP contained 8 α-helixes, 7 β-strands, and 20 coils. The SeCP had 15 potential antigenic epitopes and 19 HLA-I restricted epitopes. Based on the phylogenetic analysis of SeCP, S. erinaceieuropaei has the closest evolutionary status with S. mansonoides. SeCP was a kind of proteolytic enzyme with a variety of biological functions and its antigenic epitopes could provide important insights on the diagnostic antigens and target molecular of antisparganum drugs. Li Na Liu, Jing Cui, Xi Zhang, Tong Wei, Peng Jiang, and Zhong Quan Wang Copyright © 2013 Li Na Liu et al. All rights reserved. Proteomics Approach to the Study of Cattle Tick Adaptation to White Tailed Deer Mon, 02 Dec 2013 15:15:06 +0000 Cattle ticks, Rhipicephalus (Boophilus) microplus, are a serious threat to animal health and production. Some ticks feed on a single host species while others such as R. microplus infest multiple hosts. White tailed deer (WTD) play a role in the maintenance and expansion of cattle tick populations. However, cattle ticks fed on WTD show lower weight and reproductive performance when compared to ticks fed on cattle, suggesting the existence of host factors that affect tick feeding and reproduction. To elucidate these factors, a proteomics approach was used to characterize tick and host proteins in R. microplus ticks fed on cattle and WTD. The results showed that R. microplus ticks fed on cattle have overrepresented tick proteins involved in blood digestion and reproduction when compared to ticks fed on WTD, while host proteins were differentially represented in ticks fed on cattle or WTD. Although a direct connection cannot be made between differentially represented tick and host proteins, these results suggested that differentially represented host proteins together with other host factors could be associated with higher R. microplus tick feeding and reproduction observed in ticks fed on cattle. Marina Popara, Margarita Villar, Lourdes Mateos-Hernández, Isabel G. Fernández de Mera, and José de la Fuente Copyright © 2013 Marina Popara et al. All rights reserved. Treatment of Chagas Cardiomyopathy Sun, 24 Nov 2013 13:55:10 +0000 Chagas' disease (ChD), caused by the protozoa Trypanosoma cruzi (T. cruzi), was discovered and described by the Brazilian physician Carlos Chagas in 1909. After a century of original description, trypanosomiasis still brings much misery to humanity and is classified as a neglected tropical disease prevalent in underdeveloped countries, particularly in South America. It is an increasing worldwide problem due to the number of cases in endemic areas and the migration of infected subjects to more developed regions, mainly North America and Europe. Despite its importance, chronic chagas cardiomyopathy (CCC) pathophysiology is yet poorly understood, and independently of its social, clinical, and epidemiological importance, the therapeutic approach of CCC is still transposed from the knowledge acquired from other cardiomyopathies. Therefore, the objective of this review is to describe the treatment of Chagas cardiomyopathy with emphasis on its peculiarities. Fernando A. Botoni, Antonio Luiz P. Ribeiro, Carolina Coimbra Marinho, Marcia Maria Oliveira Lima, Maria do Carmo Pereira Nunes, and Manoel Otávio C. Rocha Copyright © 2013 Fernando A. Botoni et al. All rights reserved. The Hidden Burden of Trichinellosis in Vietnam: A Postoutbreak Epidemiological Study Thu, 07 Nov 2013 08:42:59 +0000 A cross-sectional study was conducted in Muong Lat town (Thanh Hoa province, North Vietnam), following the confirmed diagnosis of trichinellosis in six patients from that town who had eaten hunted wild boar meat during the Vietnamese lunar year celebration. All inhabitants who declared to have eaten undercooked or raw wild boar meat at the celebration and showed at least one clinical symptom compatible with trichinellosis were included in the study and blood sampled. Anti-Trichinella IgG were determined by ELISA and Western Blot. Seropositive persons were given appropriate albendazole treatment and were followed up. A total of 100 inhabitants met the inclusion criteria. Among these, 30 (30%) had antibodies to Trichinella. Serologically confirmed cases had fever (90.0%), myalgia (86.7%), facial oedema (63.3%), diarrhoea (53.3%), and pain of the masseter muscles (43.3%). Eosinophilia was detected in 83.3% of these individuals. Clinical symptoms resolved in all patients during albendazole treatment. The results suggest that only a proportion of the trichinellosis cases had sought health care during the outbreak. There is a need to implement surveillance and better diagnosis for trichinellosis and to set up educational programs to prevent infection in North Vietnam. Nga Vu Thi, Dung Do Trung, Amber Litzroth, Nicolas Praet, Huong Nguyen Thu, Hien Nguyen Thu, Hung Nguyen Manh, and Pierre Dorny Copyright © 2013 Nga Vu Thi et al. All rights reserved. Helminth Protein Vaccine Induced Follicular T Helper Cell for Enhancement of Humoral Immunity against Schistosoma japonicum Wed, 06 Nov 2013 13:41:21 +0000 Protein vaccines combined with adjuvants have been widely used to induce immune responses, especially the humoral immune response, against molecular targets including parasites. Follicular T helper (Tfh) cells are the specialized providers of B-cell help, however, the induction of Tfh cells in protein vaccination has been rarely studied. Here, we report that the Schistosoma japonicum recombinant protein (SjGST-32) combined with tacrolimus (FK506) augmented the induction of Tfh cells, which expressed the canonical markers CXCR5, BCL6, and IL-21, and enhanced the humoral immune responses in BALB/c mice. Furthermore, the expression of IL-21R on germinal center (GC) B cells and memory B cells increased in immunized mice, which indicated that IL-21 from the induced Tfh cells interacted with IL-21R for activation of B cells and maintenance of long-lived humoral immunity. Our results suggest that helminth protein vaccine combined with FK506 induces Tfh cell for stimulating humoral immune responses and inducing long-lived humoral immunity. Jingyao Zhang, Wenjuan Gao, Qirui Guo, Bobo Huang, Bin Wang, Guoliang Xia, and Youmin Kang Copyright © 2013 Jingyao Zhang et al. All rights reserved. Performance of Glutamate Dehydrogenase and Triose Phosphate Isomerase Genes in the Analysis of Genotypic Variability of Isolates of Giardia duodenalis from Livestocks Wed, 06 Nov 2013 08:10:46 +0000 Giardia duodenalis is a small intestinal protozoan parasite of several terrestrial vertebrates. This work aims to assess the genotypic variability of Giardia duodenalis isolates from cattle, sheep and pigs in the Southeast of Brazil, by comparing the standard characterization between glutamate dehydrogenase (gdh) and triose phosphate isomerase (tpi) primers. Fecal samples from the three groups of animals were analyzed using the zinc sulphate centrifugal flotation technique. Out of 59 positive samples, 30 were from cattle, 26 from sheep and 3 from pigs. Cyst pellets were stored and submitted to PCR and nested-PCR reactions with gdh and tpi primers. Fragment amplification of gdh and tpi genes was observed in 25 (42.4%) and 36 (61.0%) samples, respectively. Regarding the sequencing, 24 sequences were obtained with gdh and 20 with tpi. For both genes, there was a prevalence of E specific species assemblage, although some isolates have been identified as A and B, by the tpi sequencing. This has also shown a larger number of heterogeneous sequences, which have been attribute to mixed infections between assemblages B and E. The largest variability of inter-assemblage associated to the frequency of heterogeneity provided by tpi sequencing reinforces the polymorphic nature of this gene and makes it an excellent target for studies on molecular epidemiology. Natália M. N. Fava, Rodrigo M. Soares, Luana A. M. Scalia, Evanguedes Kalapothakis, Isabella F. Pena, Carlos U. Vieira, Elaine S. M. Faria, Maria J. Cunha, Talles R. Couto, and Márcia Cristina Cury Copyright © 2013 Natália M. N. Fava et al. All rights reserved. Molecular Identification and Polymorphism Determination of Cutaneous and Visceral Leishmaniasis Agents Isolated from Human and Animal Hosts in Iran Mon, 28 Oct 2013 14:32:54 +0000 Amplification of internal transcript spacer 1 of ribosomal RNA (ITS1-RNA) gene followed by RFLP analysis and sequencing was used to identify the causing agents of cutaneous and visceral leishmaniasis (CL and VL) in humans and animal reservoir hosts from various geographical areas in Iran. We also used random amplified polymorphic DNA (RAPD-PCR) to obtain polymorphisms among isolates of Leishmania spp. Totally, 362 suspected human and animal cases including 173 CL, 49 VL, 60 rodents, and 80 domestic dogs were examined for Leishmania infection. From 112 culture-positive samples prepared from CL cases, 75 (67%) were infected with L. major and 37 (33%) with L. tropica. Of the 60 rodents examined, 25 (41.6%) harbored the Leishmania infection; 21 were infected with L. major and 4 with L. turanica. From 49 suspected VL, 29 were positive by direct agglutination test (DAT), whereas microscopy detected parasite in bone marrow of 25 and culture in 28 of the patients. Two VL patients were infected with L. tropica and 26 with L. infantum. Of the 80 domestic dogs, 56 showed anti-Leishmania antibodies with DAT. Of these, 55 were positive by both microscopy and culture. Molecular identity, obtained only for 47 samples, revealed L. infantum in 43 and L. tropica in 4 dogs. The polymorphisms among L. tropica and L. major isolates were 3.6% and 7.3%; the rate among human and canine VL isolates was 2.8% and 9.8%, respectively. Our results showed that at least four different Leishmania species with various polymorphisms circulate among humans and animal hosts in Iran. Homa Hajjaran, Mehdi Mohebali, Setareh Mamishi, Farzaneh Vasigheh, Mohammad Ali Oshaghi, Saied Reza Naddaf, Aref Teimouri, Gholam Hossein Edrissian, and Zabiholah Zarei Copyright © 2013 Homa Hajjaran et al. All rights reserved. Genetically Distinct Glossina fuscipes fuscipes Populations in the Lake Kyoga Region of Uganda and Its Relevance for Human African Trypanosomiasis Wed, 02 Oct 2013 09:25:30 +0000 Tsetse flies (Glossina spp.) are the sole vectors of Trypanosoma brucei—the agent of human (HAT) and animal (AAT) trypanosomiasis. Glossina fuscipes fuscipes (Gff) is the main vector species in Uganda—the only country where the two forms of HAT disease (rhodesiense and gambiense) occur, with gambiense limited to the northwest. Gff populations cluster in three genetically distinct groups in northern, southern, and western Uganda, respectively, with a contact zone present in central Uganda. Understanding the dynamics of this contact zone is epidemiologically important as the merger of the two diseases is a major health concern. We used mitochondrial and microsatellite DNA data from Gff samples in the contact zone to understand its spatial extent and temporal stability. We show that this zone is relatively narrow, extending through central Uganda along major rivers with south to north introgression but displaying no sex-biased dispersal. Lack of obvious vicariant barriers suggests that either environmental conditions or reciprocal competitive exclusion could explain the patterns of genetic differentiation observed. Lack of admixture between northern and southern populations may prevent the sympatry of the two forms of HAT disease, although continued control efforts are needed to prevent the recolonization of tsetse-free regions by neighboring populations. Richard Echodu, Mark Sistrom, Chaz Hyseni, John Enyaru, Loyce Okedi, Serap Aksoy, and Adalgisa Caccone Copyright © 2013 Richard Echodu et al. All rights reserved. Beauveria bassiana Strains for Biological Control of Cosmopolites sordidus (Germ.) (Coleoptera: Curculionidae) in Plantain Thu, 26 Sep 2013 12:08:37 +0000 The objective of this study was to select strains of Beauveria bassiana for controlling Cosmopolites sordidus (Germ.) in plantain farms (cv. Terra) of the “Recôncavo” and southern regions in the state of Bahia, Brazil. The virulence of 32 B. bassiana isolates against C. sordidus was determined under laboratory conditions. Three isolates (CNPMF 407, CNPMF 218, and CNPMF 416) were selected for evaluation under field conditions in plantations located in the counties of Mutuípe and Wenceslau Guimarães. Population of C. sordidus was estimated every 15 days by using pseudostem traps. The efficiency of the three strains of B. bassiana was compared to chemical control (carbofuran, 4g/trap) and absence of control. Carbofuran caused around 90% of adult mortality after 12 months, with a reduction in the population of C. sordidus since the first evaluation. A low number of trapped insects was observed in the fungus-treated plots, suggesting the efficiency of the isolates in controlling the C. sordidus population. The strain CNPMF 218 was the most efficient in controlling C. sordidus adults in both locations, causing around 20% mortality, leading to 40% population size reduction after 12 months. Marilene Fancelli, Alex Batista Dias, Italo Delalibera Júnior, Sandra Cerqueira de Jesus, Antonio Souza do Nascimento, Sebastião de Oliveira e Silva, Ranulfo Correa Caldas, and Carlos Alberto da Silva Ledo Copyright © 2013 Marilene Fancelli et al. All rights reserved. Comparison of a Clinic-Based ELISA Test Kit with the Immunofluorescence Antibody Test for Assaying Leishmania infantum Antibodies in Dogs Thu, 26 Sep 2013 08:37:44 +0000 This study compares a rapid Immunospecific Kalazar Canine Rapid Spot IF with the gold standard test (indirect fluorescent antibody test (IFAT)) for detection of Leishmania infantum specific IgG serum antibodies in naturally exposed dogs. Serum samples were obtained from 89 healthy dogs and dogs affected by canine leishmaniosis (CanL). IgG-IFAT titers ≥80 were considered positive. Anti-L. infantum IgG antibodies were found in 54 samples with titers ranging from 1 : 80 to 1 : 5120. The performance of the rapid Immunospecific Kalazar was evaluated using a ROC curve. The area under the ROC curve of 0.957 was significantly different from 0.5 (), and therefore it can be concluded that the rapid Immunospecific Kalazar has the ability to distinguish canine sera with and without L. infantum IgG. The best performance of the test was at a cutoff >0 (sensitivity 92.6%, specificity 97%). The test can be used for disease screening if the cutoff is >0 (highest sensitivity, 92.6%) and is recommended as confirmatory test for the presence of L. infantum IgG antibodies if the cutoff is set >2 (highest specificity, 100%). Daniela Proverbio, Eva Spada, Luciana Baggiani, Giada Bagnagatti De Giorgi, and Roberta Perego Copyright © 2013 Daniela Proverbio et al. All rights reserved. Actin, RhoA, and Rab11 Participation during Encystment in Entamoeba invadens Tue, 24 Sep 2013 17:48:57 +0000 In the genus Entamoeba, actin reorganization is necessary for cyst differentiation; however, its role is still unknown. The aim of this work was to investigate the role of actin and encystation-related proteins during Entamoeba invadens encystation. Studied proteins were actin, RhoA, a small GTPase involved through its effectors in the rearrangement of the actin cytoskeleton; Rab11, a protein involved in the transport of encystation vesicles; and enolase, as an encystment vesicles marker. Results showed a high level of polymerized actin accompanied by increased levels of RhoA-GTP during cell rounding and loss of vacuoles. Cytochalasin D, an actin polymerization inhibitor, and Y27632, an inhibitor of RhoA activity, reduced encystment in 80%. These inhibitors also blocked cell rounding, disposal of vacuoles, and the proper formation of the cysts wall. At later times, F-actin and Rab11 colocalized with enolase, suggesting that Rab11 could participate in the transport of the cyst wall components through the F-actin cytoskeleton. These results suggest that actin cytoskeleton rearrangement is playing a decisive role in determining cell morphology changes and helping with the transport of cell wall components to the cell surface during encystment of E. invadens. M. Herrera-Martínez, V. I. Hernández-Ramírez, A. E. Lagunes-Guillén, B. Chávez-Munguía, and P. Talamás-Rohana Copyright © 2013 M. Herrera-Martínez et al. All rights reserved. Molecular Identification of Ancylostoma caninum Isolated from Cats in Southern China Based on Complete ITS Sequence Mon, 23 Sep 2013 16:24:01 +0000 Ancylostoma caninum is a blood-feeding parasitic intestinal nematode which infects dogs, cats, and other mammals throughout the world. A highly sensitive and species-specific PCR-RFLP technique was utilised to detect the prevalence of A. caninum in cats in Guangzhou, southern China. Of the 102 fecal samples examined, the prevalence of A. caninum in cats was 95.1% and 83.3% using PCR-RFLP and microscopy, respectively. Among them, the prevalence of single hookworm infection with A. caninum was 54.90%, while mixed infections with both A. caninum and A. ceylanicum were 40.20%. Comparative analysis of three complete ITS sequences obtained from cat-derived A. caninum showed the same length (738 bp) as that of dog-derived A. caninum. However, the sequence variation range was 98.6%–100%, where only one cat isolate (M63) showed 100% sequence similarity in comparison with two dog-derived A. caninum isolates (AM850106, EU159416) in the same studied area. The phylogenetic tree revealed A. caninum derived from both cats and dogs in single cluster. Results suggest that cats could be the main host of A. caninum in China, which may cause cross-infection between dogs and cats in the same area. Yuanjia Liu, Guochao Zheng, Muhamd Alsarakibi, Xinheng Zhang, Wei Hu, Pengyun Lu, Liqin Lin, Liping Tan, Qin Luo, and Guoqing Li Copyright © 2013 Yuanjia Liu et al. All rights reserved. Trypanosoma evansi and Surra: A Review and Perspectives on Transmission, Epidemiology and Control, Impact, and Zoonotic Aspects Wed, 18 Sep 2013 14:28:49 +0000 This paper reviews the transmission modes of Trypanosoma evansi. Its worldwide distribution is attributed to mechanical transmission. While the role of tabanids is clear, we raise questions on the relative role of Haematobia sp. and the possible role of Stomoxys sp. in delayed transmission. A review of the available trypanocidal drugs and their efficacy in various host species is useful for understanding how they interact in disease epidemiology, which is complex. Although there are similarities with other mechanically transmitted trypanosomes, T. evansi has a more complex epidemiology due to the diversity of its hosts and vectors. The impact of clinical and subclinical disease is difficult to establish. A model was developed for buffaloes in the Philippines, which could be transferred to other places and livestock systems. Since Trypanosoma evansi was reported in humans, further research is required to investigate its zoonotic potential. Surra remains a potentially emerging disease that is a threat to Australia, Spain, and France. A number of questions about the disease have yet to be resolved. This brief review of the basic knowledge of T. evansi suggests that there is renewed interest in the parasite, which is spreading and has a major economic impact. Marc Desquesnes, Alan Dargantes, De-Hua Lai, Zhao-Rong Lun, Philippe Holzmuller, and Sathaporn Jittapalapong Copyright © 2013 Marc Desquesnes et al. All rights reserved. Cystic Echinococcosis in a Single Tertiary Care Center in Rome, Italy Sat, 14 Sep 2013 11:28:48 +0000 Background. Cystic echinococcosis (CE) is a chronic, clinically complex, and neglected disease. Its prevalence in Italy, a country of medium to high endemicity, remains poorly defined, as notification has long ceased to be mandatory. Methods. We set up a retrospective cohort study involving all CE patients followed at our institute between January 2005 and December 2012. Demographical and clinical features were recorded and analyzed. Results. CE was found in 28 patients (64.3%), mostly Italians from the central regions (50%), followed by subjects from the islands (33.3%) and Southern Italy (16.7%). Their median age was 45 years (IQR: 38.5–66.5), with Eastern Europeans being significantly younger (28 years, IQR: 19–39) than other patients (). A total of 149 cysts, mostly with hepatic localization (96%), were described. Based on the WHO classification, the cysts were mainly small (80.5%) and active (CE1 (73.8%); CE2 (7.4%)). Active cysts were more common in Eastern Europeans (85.7%) than Italians (66.7%). Conclusion. Our data confirm CE occurrence in Italy. We emphasize the importance to have a national CE registry, opportunely recently introduced. This is essential to assess CE prevalence in this country, implement appropriate control measures, and improve patient management. Linda Petrone, Gilda Cuzzi, Lidia Colace, Giuseppe Maria Ettorre, Elisa Busi-Rizzi, Vincenzo Schininà, Leopoldo Pucillo, Claudio Angeletti, Stefania Pane, Antonino Di Caro, Eugenio Bordi, Enrico Girardi, Edoardo Pozio, Angela Corpolongo, Antonella Teggi, Enrico Brunetti, and Delia Goletti Copyright © 2013 Linda Petrone et al. All rights reserved. Fitness Cost of Litomosoides sigmodontis Filarial Infection in Mite Vectors; Implications of Infected Haematophagous Arthropod Excretory Products in Host-Vector Interactions Mon, 09 Sep 2013 14:54:01 +0000 Filariae are a leading cause of infections which are responsible for serious dermatological, ocular, and vascular lesions. Infective third stage larvae (L3) are transmitted through the bite of a haematophagous vector. Litomosoides sigmodontis is a well-established model of filariasis in the mouse, with the vector being the mite Ornithonyssus bacoti. The aim of the study was to analyse the filarial infection in mites to determine the consequences of filarial infection in the blood-feeding and the reproduction of mites as well as in the regulation of vector-induced inflammation in the mouse skin. Firstly, L3 are unevenly distributed throughout the host population and the majority of the population harbours a moderate infection (1 to 6 L3). Filarial infection does not significantly affect the probing delay for blood feeding. The number of released protonymphs is lower in infected mites but is not correlated with the L3 burden. Finally, induced excreted proteins from infected mites but not from uninfected mites stimulate TNF-α and the neutrophil-chemoattractant KC production by antigen-presenting cells (APCs). Altogether, these results describe the modification of the mite behavior under filarial infection and suggest that the immunomodulatory capacity of the mite may be modified by the presence of the parasite, hindering its defensive ability towards the vertebrate host. Adélaïde Nieguitsila, Roger Frutos, Catherine Moulia, Nathaly Lhermitte-Vallarino, Odile Bain, Laurent Gavotte, and Coralie Martin Copyright © 2013 Adélaïde Nieguitsila et al. All rights reserved. Comparative Analyses of the β-Tubulin Gene and Molecular Modeling Reveal Molecular Insight into the Colchicine Resistance in Kinetoplastids Organisms Sun, 08 Sep 2013 14:04:54 +0000 Differential susceptibility to microtubule agents has been demonstrated between mammalian cells and kinetoplastid organisms such as Leishmania spp. and Trypanosoma spp. The aims of this study were to identify and characterize the architecture of the putative colchicine binding site of Leishmania spp. and investigate the molecular basis of colchicine resistance. We cloned and sequenced the β-tubulin gene of Leishmania (Viannia) guyanensis and established the theoretical 3D model of the protein, using the crystallographic structure of the bovine protein as template. We identified mutations on the Leishmania  β-tubulin gene sequences on regions related to the putative colchicine-binding pocket, which generate amino acid substitutions and changes in the topology of this region, blocking the access of colchicine. The same mutations were found in the β-tubulin sequence of kinetoplastid organisms such as Trypanosoma cruzi, T. brucei, and T. evansi. Using molecular modelling approaches, we demonstrated that conformational changes include an elongation and torsion of an α-helix structure and displacement to the inside of the pocket of one β-sheet that hinders access of colchicine. We propose that kinetoplastid organisms show resistance to colchicine due to amino acids substitutions that generate structural changes in the putative colchicine-binding domain, which prevent colchicine access. Luis Luis, María Luisa Serrano, Mariana Hidalgo, and Alexis Mendoza-León Copyright © 2013 Luis Luis et al. All rights reserved. Seroprevalence, Detection of DNA in Blood and Milk, and Genotyping of Toxoplasma gondii in a Goat Population in Italy Wed, 04 Sep 2013 08:08:50 +0000 Toxoplasma gondii is the causative agent of a major zoonosis with cosmopolitan distribution and is known to be transmitted mainly by the ingestion of undercooked or raw animal products. Drinking unpasteurized goat’s milk is a risk factor associated with human toxoplasmosis. However, very little is known about the excretion of DNA in goat milk. Aim of the present study was to determine the seroprevalence of T. gondii infection using a modified agglutination test (MAT), to detect T. gondii DNA by nested-PCR (n-PCR) in samples of blood and milk from seropositive goats, and to genotype DNA isolates using 11 molecular markers in 127 adult lactating goats from 6 farms in Italy. Positive MAT results were found in 60.6% of goats while 13% of blood and milk samples from seropositive goats were positive to n-PCR. A kappa coefficient of 1 indicated a perfect agreement between blood and milk n-PCR. Genetic characterization of isolates revealed the occurrence of genotype III (), genotype I (), and atypical genotypes with hints for genotype I (). Our results suggest that the risk of excretion of Toxoplasma tachyzoites might frequently occur in milk of seropositive goats testing positive to n-PCR on blood. Francesca Mancianti, Simona Nardoni, Carlo D'Ascenzi, Francesca Pedonese, Linda Mugnaini, Filomena Franco, and Roberto Papini Copyright © 2013 Francesca Mancianti et al. All rights reserved. Exploring the Possible Role of Lysine Acetylation on Entamoeba histolytica Virulence: A Focus on the Dynamics of the Actin Cytoskeleton Sun, 01 Sep 2013 15:55:37 +0000 Cytoskeleton remodeling can be regulated, among other mechanisms, by lysine acetylation. The role of acetylation on cytoskeletal and other proteins of Entamoeba histolytica has been poorly studied. Dynamic rearrangements of the actin cytoskeleton are crucial for amebic motility and capping formation, processes that may be effective means of evading the host immune response. Here we report the possible effect of acetylation on the actin cytoskeleton dynamics and in vivo virulence of E. histolytica. Using western blot, immunoprecipitation, microscopy assays, and in silico analysis, we show results that strongly suggest that the increase in Aspirin-induced cytoplasm proteins acetylation reduced cell movement and capping formation, likely as a consequence of alterations in the structuration of the actin cytoskeleton. Additionally, intrahepatic inoculation of Aspirin-treated trophozoites in hamsters resulted in severe impairment of the amebic virulence. Taken together, these results suggest an important role for lysine acetylation in amebic invasiveness and virulence. L. López-Contreras, V. I. Hernández-Ramírez, A. E. Lagunes-Guillén, Sarita Montaño, B. Chávez-Munguía, B. Sánchez-Ramírez, and P. Talamás-Rohana Copyright © 2013 L. López-Contreras et al. All rights reserved. A Comparative Histopathology, Serology and Molecular Study, on Experimental Ocular Toxocariasis by Toxocara cati in Mongolian Gerbils and Wistar Rats Thu, 29 Aug 2013 15:22:07 +0000 The aim of this study was to compare the performance of three in-house diagnostic tests, that is, histopathology, enzyme-linked immunosorbent assay (ELISA), and polymerase chain reaction (PCR), for the diagnosis after experimental infection with Toxocara cati. Twenty Mongolian gerbils and Wistar rats were divided into ten groups (/group). Toxocara cati infections were established in Mongolian gerbils and Wistar rats by administering doses of 240 and 2500 embryonated Toxocara cati eggs by gavage, respectively. Tissue sections were stained with Haematoxylin and Eosin and observed under the light microscope. Sera and vitreous fluid collected from separate infected groups were tested against Toxocara cati antigens, for 92 days postinfection. Genomic DNA was extracted from formalin-fixed paraffin-embedded (FFPE) blocks, and aqueous fluids belong to the animals. The histopathology test gave negative results among the groups of animals examined between 5 and 92 days postinfection. The ELISA results showed that anti-Toxocara antibodies have risen between 7 and 61 days postinfection in sera and vitreous fluid in the animals infected, respectively. Analysis of PCR products revealed positive band (660 bp) in the orbital tissue infected Mongolian gerbils at 5 days postinfection. Of the three evaluated methods, the PCR could be recommended for scientific and laboratory diagnoses of toxocariasis in experimentally infected animals. Mohammad Zibaei, Seyed Mahmoud Sadjjadi, Mehdi Karamian, Shoji Uga, Ahmad Oryan, and Seyed Hamidreza Jahadi-Hosseini Copyright © 2013 Mohammad Zibaei et al. All rights reserved. Molecular Prevalence of Babesia bigemina and Trypanosoma evansi in Dairy Animals from Punjab, India, by Duplex PCR: A Step Forward to the Detection and Management of Concurrent Latent Infections Thu, 29 Aug 2013 11:59:25 +0000 Specific duplex polymerase chain reaction (PCR) was employed on 411 (386 cattle and 25 buffaloes) blood samples of dairy animals from 9 districts of Punjab, India, for simultaneous detection of Babesia bigemina and Trypanosoma evansi. The results were compared and correlated with conventional Giemsa stained thin blood smear (GSTBS) examination and haematological alterations to know the clinical status and pathogenicity of infections. The Bg3/Bg4 and TR3/TR4 primers were used in duplex PCR for B. bigemina and T. evansi amplified products of 689 bp and 257 bp, respectively. The overall prevalence by duplex PCR was found to be 36.49, 2.43, and 3.41% for T. evansi, B. bigemina, and dual infection, respectively. A more significant difference was observed for dual infection status () as compared to T. evansi () and B. bigemina () among various districts under study. A very low prevalence of T. evansi (0.73%) and B. bigemina (0.48%) was seen by GSTBS. The highly sensitive, specific, and cost-effective duplex PCR was able to detect latent T. evansi and B. bigemina infection in cattle and buffaloes. Haematological evaluation revealed marked pathology in B. bigemina infected group and in dual infected group in contrast to that infected with T. evansi alone. Amrita Sharma, Lachhman Das Singla, Ashuma Tuli, Paramjit Kaur, Balwinder Kaur Batth, Mohammed Javed, and Prayag Dutt Juyal Copyright © 2013 Amrita Sharma et al. All rights reserved. Anticestodal Activity of Endophytic Pestalotiopsis sp. on Protoscoleces of Hydatid Cyst Echinococcus granulosus Sun, 25 Aug 2013 08:10:40 +0000 Surgery is still the main treatment in hydatidosis caused by Echinococcus, which is a global health problem in human and animals. So, there is need for some natural protoscolicidal agents for instillation to prevent their reoccurrence at therapeutic doses. In this present investigation, anticestodal activity of one of the endophytic fungi Pestalotiopsis sp. from Neem plant was observed on protoscoleces of hydatid cysts of Echinococcus granulosus. Viability of protoscoleces was confirmed by 0.1% aqueous eosin red stain method, where mortality was observed at different concentrations with respect to time. An average anticestodal activity was observed with different endophytic fungal strains, that is, Nigrospora (479 ± 2.9), Colletotrichum (469 ± 25.8), Fusarium (355 ± 14.5), and Chaetomium (332 ± 28.3) showing 64 to 70% protoscolicidal activity, except Pestalotiopsis sp. (581 ± 15.0), which showed promising scolicidal activity up to 97% mortality just within 30 min of incubation. These species showed significant reduction in viability of protoscoleces. This is the first report on the scolicidal activity of endophytic Pestalotiopsis sp. We conclude that ultrastructural changes in protoscoleces were due to endophytic extract suggesting that there may be some bioactive compounds that have selective action on the tegument layer of protoscoleces. As compared with that of standard drug used, endophytic species of Neem plant shows significant anticestodal activity. Vijay C. Verma, Mayank Gangwar, Madhu Yashpal, and Gopal Nath Copyright © 2013 Vijay C. Verma et al. All rights reserved. Anthelmintic Effects of Alkylated Diamines and Amino Alcohols against Schistosoma mansoni Tue, 20 Aug 2013 08:30:59 +0000 Polyamines are substances involved in many aspects of cell growth, division, and differentiation. Because of the metabolic differences between host cells and parasite cells, polyamine metabolism has been considered as a potential target for the chemotherapy of parasitic diseases. The aim of this work was to evaluate the schistosomicidal activity of different N-alkylated diamines (3a–3h), amino alcohols (4a–4d), and glycosylated amino alcohols (10a–10d). Compounds were prepared by synthetic methods and submitted to in vitro evaluation against adult worms of Schistosoma mansoni. At 100 M, 3b, 3e, and 3h as well as 4a, 4b, 4d, 10a, 10b, and 10d resulted in 100% mortality of adult schistosomes. Compound 3d (12.5 to 100 M) caused the death of 100% of both male and female adult schistosomes, while 3f (12.5 to 100 M) resulted in 100% mortality of only male adult worms, whereas no mortality in female worms was observed. Compounds 3d and 3f were also able to reduce viability and decrease production of developed eggs in comparison with the negative control group. Diamines 3d and 3f may represent useful lead compounds for further optimization in order to develop new schistosomicidal agents. Fábio de Souza Fernandes, Celso O. Rezende Júnior, Tayrine Silva Fernandes, Lígia Souza da Silveira, Carlos A. M. Rezende, Mauro V. De Almeida, Renato G. de Paula, Vanderlei Rodrigues, Ademar A. Da Silva Filho, and Mara R. C. Couri Copyright © 2013 Fábio de Souza Fernandes et al. All rights reserved. Sphingomyelinase Activity of Trichomonas vaginalis Extract and Subfractions Mon, 19 Aug 2013 08:49:54 +0000 Trichomoniasis is one of the most common acute sexually transmitted curable diseases, and it is disseminated worldwide generating more than 170 million cases annually. Trichomonas vaginalis is the parasite that causes trichomoniasis and has the ability to destroy cell monolayers of the vaginal mucosa in vitro. Sphingomyelinases (SMase) are enzymes that catalyze the hydrolysis of sphingomyelin into ceramide and phosphorylcholine. Ceramide appears to be a second messenger lipid in programmed apoptosis, cell differentiation, and cell proliferation. Sphingomyelinase is probably a major source of ceramide in cells. Signal transduction mediated by ceramide leads cells to produce cytokine induced apoptosis during several inflammatory responses. SMase are also relevant toxins in several microorganisms. The main objective of this research is to identify SMase activity of T. vaginalis in the total extract (TE), P30, and S30 subfractions from brooked trophozoites. It was found that these fractions of T. vaginalis have SMase activity, which comes principally from P30 subfraction and was mainly type C. Enzymatic activity of SMase increased linearly with time and is pH dependent with two peaks by pH 5.5 and pH 7.5. The addition of manganese to the reaction mixture increased the SMase activity by 1.97. Francisco González-Salazar, Jesús N. Garza-González, Carlos E. Hernandez-Luna, Benito David Mata-Cárdenas, Pilar Carranza-Rosales, Jorge Enrique Castro-Garza, Magda Elizabeth Hernández-García, and Javier Vargas-Villarreal Copyright © 2013 Francisco González-Salazar et al. All rights reserved. Trypanosoma evansi and Surra: A Review and Perspectives on Origin, History, Distribution, Taxonomy, Morphology, Hosts, and Pathogenic Effects Mon, 19 Aug 2013 08:20:12 +0000 Trypanosoma evansi, the agent of “surra,” is a salivarian trypanosome, originating from Africa. It is thought to derive from Trypanosoma brucei by deletion of the maxicircle kinetoplastic DNA (genetic material required for cyclical development in tsetse flies). It is mostly mechanically transmitted by tabanids and stomoxes, initially to camels, in sub-Saharan area. The disease spread from North Africa towards the Middle East, Turkey, India, up to 53° North in Russia, across all South-East Asia, down to Indonesia and the Philippines, and it was also introduced by the conquistadores into Latin America. It can affect a very large range of domestic and wild hosts including camelids, equines, cattle, buffaloes, sheep, goats, pigs, dogs and other carnivores, deer, gazelles, and elephants. It found a new large range of wild and domestic hosts in Latin America, including reservoirs (capybaras) and biological vectors (vampire bats). Surra is a major disease in camels, equines, and dogs, in which it can often be fatal in the absence of treatment, and exhibits nonspecific clinical signs (anaemia, loss of weight, abortion, and death), which are variable from one host and one place to another; however, its immunosuppressive effects interfering with intercurrent diseases or vaccination campaigns might be its most significant and questionable aspect. Marc Desquesnes, Philippe Holzmuller, De-Hua Lai, Alan Dargantes, Zhao-Rong Lun, and Sathaporn Jittaplapong Copyright © 2013 Marc Desquesnes et al. All rights reserved. Epidemiology of Visceral Leishmaniasis in a Reemerging Focus of Intense Transmission in Minas Gerais State, Brazil Tue, 13 Aug 2013 13:08:15 +0000 This study was developed in the urban area of Governador Valadares, a reemerging focus of intense transmission of visceral leishmaniasis (VL) in Brazil, presenting 86 human cases of VL from 2008 to 2011. The disease prevailed in males (73.2%) with most patients between 0 and 9 years (44.1%) and a lethality rate of 16.2%. A canine survey was carried out on 16,529 domestic dogs in 35 districts in the area and it showed that 30.2% of them (4,992 dogs) were positive for VL by serum assays. Prevalence ratios for canine VL varied between 13.6% and 53.4%. The clinical exam of 343 seropositive dogs showed that 49.9% of them were considered symptomatic, with larger prevalence of canine VL being in short-furred animals (90%). The entomological survey was performed in eight districts, where 2,539 phlebotomines were captured, preferentially in the peridomicile (84.5%). Lutzomyia longipalpis was the predominant species (90%) suggesting its participation in the VL transmission in the area. The correlation between canine prevalence and L. longipalpis density was evaluated. Ricardo Andrade Barata, Jennifer Cunha Peixoto, Aline Tanure, Marcela Esteves Gomes, Estefânia Conceição Apolinário, Emerson Cotta Bodevan, Holbiano Saraiva de Araújo, Edelberto Santos Dias, and Aimara da Costa Pinheiro Copyright © 2013 Ricardo Andrade Barata et al. All rights reserved. Isolation and Identification of Free-Living Amoebae from Tap Water in Sivas, Turkey Mon, 22 Jul 2013 13:02:49 +0000 The present work focuses on a local survey of free-living amoebae (FLA) that cause opportunistic and nonopportunistic infections in humans. Determining the prevalence of FLA in water sources can shine a light on the need to prevent FLA related illnesses. A total of 150 samples of tap water were collected from six districts of Sivas province. The samples were filtered and seeded on nonnutrient agar containing Escherichia coli spread. Thirty-three (22%) out of 150 samples were found to be positive for FLA. The FLA were identified by morphology and by PCR using 18S rDNA gene. The morphological analysis and partial sequencing of the 18S rDNA gene revealed the presence of three different species, Acanthamoeba castellanii, Acanthamoeba polyphaga, and Hartmannella vermiformis. Naegleria fowleri, Balamuthia mandrillaris, or Sappinia sp. was not isolated during the study. All A. castellanii and A. polyphaga sequence types were found to be genotype T4 that contains most of the pathogenic Acanthamoeba strains. The results indicated the occurrence and distribution of FLA species in tap water in these localities of Sivas, Turkey. Furthermore, the presence of temperature tolerant Acanthamoeba genotype T4 in tap water in the region must be taken into account for health risks. Kübra Açıkalın Coşkun, Semra Özçelik, Lütfi Tutar, Nazif Elaldı, and Yusuf Tutar Copyright © 2013 Kübra Açıkalın Coşkun et al. All rights reserved. Effects of Sheep and Mouse Urine on the Growth Pattern of Leishmania major Promastigotes Mon, 15 Jul 2013 11:38:11 +0000 The protozoan parasites of the genus Leishmania are the causative agents of different clinical diseases. Fetal calf serum (FCS) is the main part and the most expensive ingredient of the Leishmania culture media. Here, the efficacies of different concentrations (1%, 2.5%, 5%, and 10%) of the filtered and autoclaved sheep and mouse urine were evaluated as a growth stimulator in Leishmania culture procedure. The results indicated that culture media enriched with the filtered sheep and mouse urine supported the growth of the parasites and can be used for cultivation of Leishmania parasites. In conclusion, this study has demonstrated an alternative low-cost medium that could be used in cultivation process of Leishmania major promastigotes. Vahid Nasiri, Gholamreza Karimi, Abdolhossein Dalimi, Habibollah Paykari, and Fatemeh Ghaffarifar Copyright © 2013 Vahid Nasiri et al. All rights reserved. Experimental Susceptibility of Gilthead Sea Bream, Sparus aurata, via Challenge with Anisakis pegreffii Larvae Thu, 11 Jul 2013 09:22:29 +0000 The endoscopic and histopathological findings detected in Sparus aurata experimentally infected with third-stage Anisakis larvae without intermediate host are evaluated and discussed. In six fish, live nematode larvae were introduced by gastroscopy into the stomach. The first observation by endoscope, 15 days after challenge, showed the presence of some larvae at the level of gastric mucosa. An explorative celioscopy, performed 60 days after challenge, showed haemorrhages and/or nodules on the gut of two fishes. Necropsy and histology demonstrated parasites in the context of the tissue changes. The finding of live nematode larvae as well as the evidence of tissue change confirm the experimental susceptibility of gilthead sea bream towards Anisakis. Fabio Marino, Giovanni Lanteri, Annamaria Passantino, Carmelo De Stefano, Antonella Costa, Gabriella Gaglio, and Francesco Macrì Copyright © 2013 Fabio Marino et al. All rights reserved. Effect of Experimental Infection with Haemonchus contortus on Parasitological and Local Cellular Responses in Resistant and Susceptible Young Creole Goats Thu, 11 Jul 2013 08:19:00 +0000 This study was carried out to evaluate the relationships of cellular changes in the abomasal mucosa and parasitological parameters, by comparing resistant and susceptible young Creole goats (kids) after experimental infection with Haemonchus contortus. The kids were infected over 2 periods (challenges 1 and 2) of 7 and 6 weeks, respectively. Fecal egg count (FEC), blood eosinophilia, packed cell volume (PCV), and body weight were weekly monitored. At the end of both challenges a subgroup of kids was slaughtered for nematode burden measurements and analysis of inflammatory cell infiltration in the abomasal mucosa. The average daily gain was higher in resistant kids after both challenges. Blood eosinophilia and FEC were higher in susceptible kids after both challenges. The number of immature worms and the means of female length were lower after challenge 2 whatever the genetic status. No differences were observed in the eosinophil and mononuclear cell infiltration between challenges 1 and 2 and resistant and susceptible kids. Globule leukocyte infiltration was found higher after the challenge 1 in resistant kids. This effect of the genetic status on globule leukocytes counts but not on the other inflammatory cell highlights the need for further study on the functional activity of these cell populations. J. C. Bambou, T. Larcher, W. Ceï, P. J. Dumoulin, and N. Mandonnet Copyright © 2013 J. C. Bambou et al. All rights reserved. Comparative Evaluation of Infected and Noninfected Amblyomma triste Ticks with Rickettsia parkeri, the Agent of an Emerging Rickettsiosis in the New World Mon, 08 Jul 2013 15:09:18 +0000 The distribution of Rickettsia parkeri in South America has been associated with Amblyomma triste ticks. The present study evaluated under laboratory conditions two colonies of A. triste: one started from engorged females that were naturally infected by R. parkeri (designated as infected group); the other started from noninfected females (designated as control group). Both colonies were reared in parallel for five consecutive generations. Tick-naïve domestic rabbits were used for feeding of each tick stage and generation. R. parkeri was preserved by transstadial maintenance and transovarial transmission in A. triste ticks for five consecutive generations, because all tested larvae, nymphs, and adults from the infected group were shown by PCR to contain rickettsial DNA. All rabbits infested by larvae, nymphs, and adults from the infected group seroconverted, indicating that these tick stages were all vector competent for R. parkeri. Expressive differences in mortality rates were observed between engorged nymphs from the infected and control groups, as indicated by 65.9% and 92.4% molting success, respectively. Our results indicate that A. triste can act as a natural reservoir for R. parkeri. However, due to deleterious effect caused by R. parkeri on engorged nymphs, amplifier vertebrate hosts might be necessary for natural long-term maintenance of R. parkeri in A. triste. F. A. Nieri-Bastos, M. P. J. Szabó, R. C. Pacheco, J. F. Soares, H. S. Soares, J. Moraes-Filho, R. A. Dias, and M. B. Labruna Copyright © 2013 F. A. Nieri-Bastos et al. All rights reserved. Genetic Variation between Biomphalaria alexandrina Snails Susceptible and Resistant to Schistosoma mansoni Infection Wed, 26 Jun 2013 10:58:58 +0000 Much effort has been made to control schistosomiasis infection in Egypt. However, enduring effects from such strategies have not yet been achieved. In this study, we sought to determine the genetic variability related to the interaction between Biomphalaria alexandrina snails and Schistosoma mansoni. Using RAPD-PCR with eight (10 mers) random primers, we were able to determine the polymorphic markers that differed between snails susceptible and resistant to Schistosoma mansoni infection using five primers out of the eight. Our results suggest that the RAPD-PCR technique is an efficient means by which to compare genomes and to detect genetic variations between schistosomiasis intermediate hosts. The RAPD technique with the above-noted primers can identify genomic markers that are specifically related to the Biomphalaria alexandrina/Schistosoma mansoni relationship in the absence of specific nucleotide sequence information. This approach could be used in epidemiologic surveys to investigate genetic diversity among Biomphalaria alexandrina snails. The ability to determine resistant markers in Biomphalaria alexandrina snails could potentially lead to further studies that use refractory snails as agents to control the spread of schistosomiasis. Suzanne M. F. El-Nassery, Iman F. Abou-El-Naga, Sonia R. Allam, Eman A. Shaat, and Rasha F. M. Mady Copyright © 2013 Suzanne M. F. El-Nassery et al. All rights reserved. Border Malaria Associated with Multidrug Resistance on Thailand-Myanmar and Thailand-Cambodia Borders: Transmission Dynamic, Vulnerability, and Surveillance Tue, 25 Jun 2013 14:20:20 +0000 This systematic review elaborates the concepts and impacts of border malaria, particularly on the emergence and spread of Plasmodium falciparum and Plasmodium vivax multidrug resistance (MDR) malaria on Thailand-Myanmar and Thailand-Cambodia borders. Border malaria encompasses any complex epidemiological settings of forest-related and forest fringe-related malaria, both regularly occurring in certain transmission areas and manifesting a trend of increased incidence in transmission prone areas along these borders, as the result of interconnections of human settlements and movement activities, cross-border population migrations, ecological changes, vector population dynamics, and multidrug resistance. For regional and global perspectives, this review analyzes and synthesizes the rationales pertaining to transmission dynamics and the vulnerabilities of border malaria that constrain surveillance and control of the world’s most MDR falciparum and vivax malaria on these chaotic borders. Adisak Bhumiratana, Apiradee Intarapuk, Prapa Sorosjinda-Nunthawarasilp, Pannamas Maneekan, and Surachart Koyadun Copyright © 2013 Adisak Bhumiratana et al. All rights reserved. Microalbuminuria and Glomerular Filtration Rate in Paediatric Visceral Leishmaniasis Sun, 23 Jun 2013 11:41:27 +0000 Visceral leishmaniasis, caused by Leishmania donovani, is a serious form of leishmaniasis and fatal if untreated. Nearly half of the VL cases are children. There are very few studies of renal function in pediatric visceral leishmaniasis. The aim of this study was to evaluate renal dysfunction by studying glomerular filtration rate (GFR), microalbuminuria, and microscopic examination of urine. Laboratory analysis was performed on blood and urine samples of 40 parasitologically confirmed pediatric VL cases. Laboratory data of urine examination showed albuminuria in 10% (4/40), white blood cells in 20% (8/40), hematuria in 10% (4/40), microalbuminuria in 37.5% (15/40), and decreased GFR in 27.5% (11/40). Renal involvement was manifested in most of the pediatric VL cases. These findings may help clinicians in decision making for safe and suitable antileishmanial treatment particularly in childhood VL. Neena Verma, Chandra Shekhar Lal, Vidyanand Rabidas, Krishna Pandey, Dharmendra Singh, Sanjay Kumar, Rakesh Bihari Verma, and Pradeep Das Copyright © 2013 Neena Verma et al. All rights reserved. Remarkable Anti-Trichomonas vaginalis Activity of Plants Traditionally Used by the Mbyá-Guarani Indigenous Group in Brazil Thu, 20 Jun 2013 17:28:31 +0000 Trichomonas vaginalis, a flagellate protozoan, is the causative agent of trichomonosis, the most common nonviral sexually transmitted disease worldwide. Taking into account the increased prevalence of metronidazole-resistant isolates, alternative drugs are essential for the successful treatment. Natural products are the source of most new drugs, and popular wisdom about the use of medicinal plants is a powerful tool in this search. In this study, the activity of 10 medicinal plants extensively used in daily life by Mbyá-Guarani indigenous group was evaluated against seven different T. vaginalis isolates. Among the aqueous extracts tested, Verbena sp. (Guachu ka'a in Mbyá-Guarani language) and Campomanesia xanthocarpa (Guavira in Mbyá-Guarani language) showed the highest activity against T. vaginalis with MIC value of 4.0 mg/mL reaching 100% of efficacy against the parasite. The kinetic growth assays showed that the extracts promoted complete growth abolishment after 4 h of incubation. In addition, the extracts tested did not promote a significant hemolytic activity against human erythrocytes. Our results show for the first time the potential activity of Verbena sp. and C. xanthocarpa against T. vaginalis. In addition, this study demonstrates that indigenous knowledge is an important source of new prototype antiprotozoal agents. Clara Lia Costa Brandelli, Patrícia de Brum Vieira, Alexandre José Macedo, and Tiana Tasca Copyright © 2013 Clara Lia Costa Brandelli et al. All rights reserved. Immunology and Cell Biology of Parasitic Diseases 2013 Mon, 17 Jun 2013 10:40:53 +0000 Luis I. Terrazas, Abhay R. Satoskar, Miriam Rodriguez-Sosa, and Jorge Morales-Montor Copyright © 2013 Luis I. Terrazas et al. All rights reserved. Proteomic Analysis of Trichinella spiralis Muscle Larval Excretory-Secretory Proteins Recognized by Early Infection Sera Thu, 13 Jun 2013 09:38:32 +0000 Although the excretory-secretory (ES) proteins of Trichinella spiralis muscle larvae are the most commonly used diagnostic antigens for trichinellosis, their main disadvantage is the false negative results during the early stage of infection and cross-reaction of their main components (43, 45, 49, and 53 kDa) with sera of patients with other helminthiasis. The aim of this study was to identify early specific diagnostic antigens in T. spiralis ES proteins with 30–40 kDa. The ES proteins were analyzed by two-dimensional electrophoresis (2-DE), and a total of approximately 150 proteins spots were detected with isoelectric point (pI) varying from 4 to 7 and molecular weight from 14 to 66 kDa. When probed with sera from infected mice at 18 days postinfection, ten protein spots with molecular weight of 30–40 kDa were recognized and identified by MALDI-TOF/TOF-MS. All of ten spots were successfully identified and characterized to correlate with five different proteins, including two potential serine proteases, one antigen targeted by protective antibodies, one deoxyribonuclease (DNase) II, and one conserved hypothetical protein. These proteins might be the early specific diagnostic antigens for trichinellosis. Li Wang, Zhong Quan Wang, Dan Dan Hu, and Jing Cui Copyright © 2013 Li Wang et al. All rights reserved. Erratum to “Sex Steroids Effects on the Molting Process of the Helminth Human Parasite Trichinella spiralis” Wed, 10 Apr 2013 14:59:01 +0000 Romel Hernández-Bello, Ricardo Ramirez-Nieto, Saé Muñiz-Hernández, Karen Nava-Castro, Lenin Pavón, Ana Gabriela Sánchez-Acosta, and Jorge Morales-Montor Copyright © 2013 Romel Hernández-Bello et al. All rights reserved. Analysis of Structures and Epitopes of Surface Antigen Glycoproteins Expressed in Bradyzoites of Toxoplasma gondii Thu, 21 Mar 2013 13:33:48 +0000 Toxoplasma gondii is a protozoan parasite capable of infecting humans and animals. Surface antigen glycoproteins, SAG2C, -2D, -2X, and -2Y, are expressed on the surface of bradyzoites. These antigens have been shown to protect bradyzoites against immune responses during chronic infections. We studied structures of SAG2C, -2D, -2X, and -2Y proteins using bioinformatics methods. The protein sequence alignment was performed by T-Coffee method. Secondary structural and functional domains were predicted using software PSIPRED v3.0 and SMART software, and 3D models of proteins were constructed and compared using the I-TASSER server, VMD, and SWISS-spdbv. Our results showed that SAG2C, -2D, -2X, and -2Y are highly homologous proteins. They share the same conserved peptides and HLA-I restricted epitopes. The similarity in structure and domains indicated putative common functions that might stimulate similar immune response in hosts. The conserved peptides and HLA-restricted epitopes could provide important insights on vaccine study and the diagnosis of this disease. Hua Cong, Min Zhang, Qingli Zhang, Jing Gong, Haizi Cong, Qing Xin, and Shenyi He Copyright © 2013 Hua Cong et al. All rights reserved. Identification of Protein Complex Associated with LYT1 of Trypanosoma cruzi Sun, 17 Mar 2013 15:14:49 +0000 To carry out the intracellular phase of its life cycle, Trypanosoma cruzi must infect a host cell. Although a few molecules have been reported to participate in this process, one known protein is LYT1, which promotes lysis under acidic conditions and is involved in parasite infection and development. Alternative transcripts from a single LYT1 gene generate two proteins with differential functions and compartmentalization. Single-gene products targeted to more than one location can interact with disparate proteins that might affect their function and targeting properties. The aim of this work was to study the LYT1 interaction map using coimmunoprecipitation assays with transgenic parasites expressing LYT1 products fused to GFP. We detected several proteins of sizes from 8 to 150 kDa that bind to LYT1 with different binding strengths. By MS-MS analysis, we identified proteins involved in parasite infectivity (trans-sialidase), development (kDSPs and histones H2A and H2B), and motility and protein traffic (dynein and α- and β-tubulin), as well as protein-protein interactions (TPR-protein and kDSPs) and several hypothetical proteins. Our approach led us to identify the LYT1 interaction profile, thereby providing insights into the molecular mechanisms that contribute to parasite stage development and pathogenesis of T. cruzi infection. C. Lugo-Caballero, G. Ballesteros-Rodea, S. Martínez-Calvillo, and Rebeca Manning-Cela Copyright © 2013 C. Lugo-Caballero et al. All rights reserved. PPAR Activation Induces M1 Macrophage Polarization via cPLA2-COX-2 Inhibition, Activating ROS Production against Leishmania mexicana Thu, 28 Feb 2013 16:04:17 +0000 Defence against Leishmania depends upon Th1 inflammatory response and, a major problem in susceptible models, is the turnoff of the leishmanicidal activity of macrophages with IL-10, IL-4, and COX-2 upregulation, as well as immunosuppressive PGE2, all together inhibiting the respiratory burst. Peroxisome proliferator-activated receptors (PPAR) activation is responsible for macrophages polarization on Leishmania susceptible models where microbicide functions are deactivated. In this paper, we demonstrated that, at least for L. mexicana, PPAR activation, mainly PPARγ, induced macrophage activation through their polarization towards M1 profile with the increase of microbicide activity against intracellular pathogen L. mexicana. PPAR activation induced IL-10 downregulation, whereas the production of proinflammatory cytokines such as TNF-α, IL-1β, and IL-6 remained high. Moreover, PPAR agonists treatment induced the deactivation of cPLA2-COX-2-prostaglandins pathway together with an increase in TLR4 expression, all of whose criteria meet the M1 macrophage profile. Finally, parasite burden, in treated macrophages, was lower than that in infected nontreated macrophages, most probably associated with the increase of respiratory burst in these treated cells. Based on the above data, we conclude that PPAR agonists used in this work induces M1 macrophages polarization via inhibition of cPLA2 and the increase of aggressive microbicidal activity via reactive oxygen species (ROS) production. J. A. Díaz-Gandarilla, C. Osorio-Trujillo, V. I. Hernández-Ramírez, and P. Talamás-Rohana Copyright © 2013 J. A. Díaz-Gandarilla et al. All rights reserved. Signal Transducer and Activator of Transcription Factor 6 Signaling Contributes to Control Host Lung Pathology but Favors Susceptibility against Toxocara canis Infection Wed, 30 Jan 2013 08:52:19 +0000 Using BALB/c mice, we have analyzed the role of STAT6-induced Th2 response in determining the outcome of experimental toxocariasis caused by embryonated eggs of the helminth parasite Toxocara canis. Following T. canis infection wild-type BALB/c mice developed a strong Th2-like response, produced high levels of IgG1, IgE, and IL-4, recruited alternatively activated macrophages, and displayed a moderate pathology in the lungs; however, they harbored heavy parasite loads in different tissues. In contrast, similarly infected BALB/c mice mounted a weak Th2-like response, did not recruit alternatively activated macrophages, displayed a severe pathology in the lungs, but efficiently controlled T. canis infection. These findings demonstrate that Th2-like response induced via STAT6-mediated signaling pathway mediates susceptibility to larval stage of T. canis. Furthermore, they also indicate that unlike most gastrointestinal helminths, immunity against larvae of T. canis is not mediated by a Th2-dominant response. Berenice Faz-López, Yadira Ledesma-Soto, Yolanda Romero-Sánchez, Elsa Calleja, Pablo Martínez-Labat, and Luis I. Terrazas Copyright © 2013 Berenice Faz-López et al. All rights reserved. Parasitic Infections: A Role for C-Type Lectins Receptors Sun, 27 Jan 2013 12:55:28 +0000 Antigen-presenting cells (APCs) sense the microenvironment through several types of receptors that recognize pathogen-associated molecular patterns. In particular, C-type lectins receptors (CLRs), which are expressed by distinct subsets of dendritic cells (DCs) and macrophages (MØs), recognize and internalize specific carbohydrate antigens in a Ca2+-dependent manner. The targeting of these receptors is becoming an efficient strategy for parasite recognition. However, relatively little is known about how CLRs are involved in both pathogen recognition and the internalization of parasites. The role of CLRs in parasite infections is an area of considerable interest because this research will impact our understanding of the initiation of innate immune responses, which influences the outcome of specific immune responses. This paper attempts to summarize our understanding of the effects of parasites’ interactions with CLRs. Alicia Vázquez-Mendoza, Julio César Carrero, and Miriam Rodriguez-Sosa Copyright © 2013 Alicia Vázquez-Mendoza et al. All rights reserved. Silencing of Entamoeba histolytica Glucosamine 6-Phosphate Isomerase by RNA Interference Inhibits the Formation of Cyst-Like Structures Tue, 22 Jan 2013 16:20:53 +0000 Encystment is an essential process in the biological cycle of the human parasite Entamoeba histolytica. In the present study, we evaluated the participation of E. histolytica Gln6Pi in the formation of amoeba cyst-like structures by RNA interference assay. Amoeba trophozoites transfected with two Gln6Pi siRNAs reduced the expression of the enzyme in 85%, which was confirmed by western blot using an anti-Gln6Pi antibody. The E. histolytica Gln6Pi knockdown with the mix of both siRNAs resulted in the loss of its capacity to form cyst-like structures (CLSs) and develop a chitin wall under hydrogen peroxide treatment, as evidenced by absence of both resistance to detergent treatment and calcofluor staining. Thus, only 5% of treated trophozoites were converted to CLS, from which only 15% were calcofluor stained. These results represent an advance in the understanding of chitin biosynthesis in E. histolytica and provide insight into the encystment process in this parasite, which could allow for the developing of new control strategies for this parasite. Hugo Aguilar-Díaz, Juan Pedro Laclette, and Julio César Carrero Copyright © 2013 Hugo Aguilar-Díaz et al. All rights reserved. Sex-Associated Expression of Co-Stimulatory Molecules CD80, CD86, and Accessory Molecules, PDL-1, PDL-2 and MHC-II, in F480+ Macrophages during Murine Cysticercosis Sun, 20 Jan 2013 08:13:37 +0000 Macrophages are critically involved in the interaction between T. crassiceps and the murine host immune system. Also, a strong gender-associated susceptibility to murine cysticercosis has been reported. Here, we examined the sex-associated expression of molecules MHC-II, CD80, CD86, PD-L1, and PD-L2 on peritoneal F4/ macrophages of BALB/c mice infected with Taenia crassiceps. Peritoneal macrophages from both sexes of mice were exposed to T. crassiceps total extract (TcEx). BALB/c Females mice recruit higher number of macrophages to the peritoneum. Macrophages from infected animals show increased expression of PDL2 and CD80 that was dependent from the sex of the host. These findings suggest that macrophage recruitment at early time points during T. crassiceps infection is a possible mechanism that underlies the differential sex-associated susceptibility displayed by the mouse gender. Cristián Togno-Peirce, Karen Nava-Castro, Luis Ignacio Terrazas, and Jorge Morales-Montor Copyright © 2013 Cristián Togno-Peirce et al. All rights reserved. The TvLEGU-1, a Legumain-Like Cysteine Proteinase, Plays a Key Role in Trichomonas vaginalis Cytoadherence Tue, 01 Jan 2013 11:52:08 +0000 The goal of this paper was to characterize a Trichomonas vaginalis cysteine proteinase (CP) legumain-1 (TvLEGU-1) and determine its potential role as a virulence factor during T. vaginalis infection. A 30-kDa band, which migrates in three protein spots (pI~6.3, ~6.5, and ~6.7) with a different type and level of phosphorylation, was identified as TvLEGU-1 by one- and two-dimensional Western blot (WB) assays, using a protease-rich trichomonad extract and polyclonal antibodies produced against the recombinant TvLEGU-1 (anti-TvLEGU-1r). Its identification was confirmed by mass spectrometry. Immunofluorescence, cell binding, and WB assays showed that TvLEGU-1 is upregulated by iron at the protein level, localized on the trichomonad surface and in lysosomes and Golgi complex, bound to the surface of HeLa cells, and was found in vaginal secretions. Additionally, the IgG and Fab fractions of the anti-TvLEGU-1r antibody inhibited trichomonal cytoadherence up to 45%. Moreover, the Aza-Peptidyl Michael Acceptor that inhibited legumain proteolytic activity in live parasites also reduced levels of trichomonal cytoadherence up to 80%. In conclusion, our data show that the proteolytic activity of TvLEGU-1 is necessary for trichomonal adherence. Thus, TvLEGU-1 is a novel virulence factor upregulated by iron. This is the first report that a legumain-like CP plays a role in a pathogen cytoadherence. Francisco Javier Rendón-Gandarilla, Lucero de los Angeles Ramón-Luing, Jaime Ortega-López, Ivone Rosa de Andrade, Marlene Benchimol, and Rossana Arroyo Copyright © 2013 Francisco Javier Rendón-Gandarilla et al. All rights reserved. Reevaluating the Role of Acanthamoeba Proteases in Tissue Invasion: Observation of Cytopathogenic Mechanisms on MDCK Cell Monolayers and Hamster Corneal Cells Tue, 01 Jan 2013 10:56:19 +0000 The morphological analysis of the cytopathic effect on MDCK cell monolayers and hamster cornea and qualitative and quantitative analyses of conditioned medium and proteases were evaluated and compared between two strains of Acanthamoeba genotype T4. Further than highlighting the biological differences found between both strains, the most important observation in this study was the fact that proteases both in total extracts and in conditioned medium are apparently not determinant in tissue destruction. An interestingly finding was that no lysis of corneal tissue was observed as it was previously suggested. These results, together with previous studies, allow us to conclude that the invasion and disruption of corneal tissue is performed by the penetration of the amoebae through cell junctions, either by the action of proteases promoting cellular separation but not by their destruction and/or a mechanical effect exerted by amoebae. Therefore, contact-dependent mechanisms in Acanthamoeba pathogenesis are more relevant than it has been previously considered. This is supported because the phagocytosis of recently detached cells as well as those attached to the corneal epithelium leads to the modification of the cellular architecture facilitating the migration and destruction of deeper layers of the corneal epithelium. Maritza Omaña-Molina, Arturo González-Robles, Lizbeth Iliana Salazar-Villatoro, Jacob Lorenzo-Morales, Ana Ruth Cristóbal-Ramos, Verónica Ivonne Hernández-Ramírez, Patricia Talamás-Rohana, Adolfo René Méndez Cruz, and Adolfo Martínez-Palomo Copyright © 2013 Maritza Omaña-Molina et al. All rights reserved. Gene Gun Bombardment with DNA-Coated Golden Particles Enhanced the Protective Effect of a DNA Vaccine Based on Thioredoxin Glutathione Reductase of Schistosoma japonicum Mon, 31 Dec 2012 11:55:01 +0000 Schistosomiasis, caused by infection with Schistosoma species, remains an important parasitic zoonosis. Thioredoxin glutathione reductase of Schistosoma japonicum (SjTGR) plays an important role in the development of the parasite and for its survival. Here we present a recombinant plasmid DNA vaccine, pVAX1/SjTGR, to estimate its protection against S. japonicum in BALB/c mice. The DNA vaccine administrated by particle bombardment induced higher protection than by intramuscular injection. All animals vaccinated with pVAX1/SjTGR developed significant specific anti-SjTGR antibodies than control groups. Moreover, animals immunized by gene gun exhibited a splenocyte proliferative response, with an increase in IFN-γ and IL-4. The recombinant plasmid administrated by gene gun achieved a medium protective efficacy of 27.83–38.83% () of worm reduction and 40.38–44.51% () of liver egg count reduction. It suggests that different modes of administering a DNA vaccine can influence the protective efficacy induced by the vaccine. Interestingly, from the enzymatic activity results, we found that worms obtained from pVAX1/SjTGR-vaccinated animals expressed lower enzymatic activity than the control group and the antibodies weakened the enzymatic activity of SjTGR in vitro, too. It implies that the high-level antibodies may contribute to the protective effects. Yan Cao, Bin Zhao, Yanhui Han, Juan Zhang, Xuezhen Li, Chunhui Qiu, Xiujuan Wu, Yang Hong, Dezhou Ai, Jiaojiao Lin, and Zhiqiang Fu Copyright © 2013 Yan Cao et al. All rights reserved. In Vitro Ovicidal and Cestocidal Effects of Toxins from Bacillus thuringiensis on the Canine and Human Parasite Dipylidium caninum Sat, 29 Dec 2012 10:12:15 +0000 Bacillus thuringiensis is a gram-positive soil-dwelling bacterium that is commonly used as a biological pesticide. This bacterium may also be used for biological control of helminth parasites in domestic animals. In this study, we evaluated the possible ovicidal and cestocidal effects of a total protein extract of B. thuringiensis native strains on the zoonotic cestode parasite of dogs, Dipylidium caninum (D. caninum). Dose and time response curves were determined by coincubating B. thuringiensis proteins at concentration ranging from 100 to 1000 μg/mL along with 4000 egg capsules of D. caninum. Egg viability was evaluated using the trypan blue exclusion test. The lethal concentration of toxins on eggs was 600 μg/ml, and the best incubation time to produce this effect was 3 h. In the adult stage, the motility and the thickness of the tegument were used as indicators of damage. The motility was inhibited by 100% after 8 hours of culture compared to the control group, while the thickness of the cestode was reduced by 34%. Conclusively, proteins of the strain GP526 of B. thuringiensis directly act upon D. caninum showing ovicidal and cestocidal effects. Thus, B. thuringiensis is proposed as a potential biological control agent against this zoonosis. Guadalupe Peña, Fortino Agustín Aguilar Jiménez, Claudia Hallal-Calleros, Jorge Morales-Montor, Víctor Manuel Hernández-Velázquez, and Fernando Iván Flores-Pérez Copyright © 2013 Guadalupe Peña et al. All rights reserved. Taenia crassiceps Infection Does Not Influence the Development of Experimental Rheumatoid Arthritis Thu, 27 Dec 2012 18:30:58 +0000 It was previously reported by our group that infection with Taenia crassiceps reduces incidence and severity of inflammatory and autoimmune experimental diseases like type 1 diabetes and experimental autoimmune encephalomyelitis. In this research, we set out to study whether infection with T. crassiceps would affect the development of experimental rheumatoid arthritis (RA). We found that mice infected with the parasite and induced with experimental RA showed similar clinical scores as the noninfected experimental RA group; systemic cytokines were not affected while anti-CII Abs were higher in the infected group. Histological evaluation showed damage in both infected and noninfected experimental RA-induced groups and although some surface molecules such as PDL-2 and MR which are associated with immunomodulatory mechanisms were upregulated in the infected and RA-induced group as compared to the noninfected RA group, they did not exert any changes in the outcome of experimental RA. Thus, we determined that infection with T. crassiceps does not influence the outcome of experimental RA. Aaxin M. Ortiz-Flores, Yadira Ledesma-Soto, Elsa A. Calleja, Miriam Rodríguez-Sosa, Imelda Juárez, and Luis I. Terrazas Copyright © 2013 Aaxin M. Ortiz-Flores et al. All rights reserved. Immunoregulation by Taenia crassiceps and Its Antigens Thu, 27 Dec 2012 15:22:42 +0000 Taenia crassiceps is a cestode parasite of rodents (in its larval stage) and canids (in its adult stage) that can also parasitize immunocompromised humans. We have studied the immune response elicited by this helminth and its antigens in mice and human cells, and have discovered that they have a strong capacity to induce chronic Th2-type responses that are primarily characterized by high levels of Th2 cytokines, low proliferative responses in lymphocytes, an immature and LPS-tolerogenic profile in dendritic cells, the recruitment of myeloid-derived suppressor cells and, specially, alternatively activated macrophages. We also have utilized the immunoregulatory capabilities of this helminth to successfully modulate autoimmune responses and the outcome of other infectious diseases. In the present paper, we review the work of others and ourselves with regard to the immune response induced by T. crassiceps and its antigens, and we compare the advances in our understanding of this parasitic infection model with the knowledge that has been obtained from other selected models. Alberto N. Peón, Arlett Espinoza-Jiménez, and Luis I. Terrazas Copyright © 2013 Alberto N. Peón et al. All rights reserved. Helminth Excreted/Secreted Antigens Repress Expression of LPS-Induced Let-7i but Not miR-146a and miR-155 in Human Dendritic Cells Thu, 27 Dec 2012 14:31:27 +0000 MicroRNAs have emerged as key regulators of immune responses. They influence immune cells' function and probably the outcome of several infections. Currently, it is largely unknown if helminth parasites and their antigens modify host microRNAs expression. The aim of this study was to explore if excreted/secreted antigens of Taenia crassiceps regulate LPS-induced miRNAs expression in human Dendritic Cells. We found that these antigens repressed LPS-let-7i induction but not mir-146a or mir-155 and this correlates with a diminished inflammatory response. This let-7i downregulation in Dendritic Cells constitutes a novel feature of the modulatory activity that helminth-derived antigens exert on their host. Luis I. Terrazas, Fausto Sánchez-Muñoz, Magaly Pérez-Miranda, Ana M. Mejía-Domínguez, Yadira Ledesma-Soto, Rafael Bojalil, and Lorena Gómez-García Copyright © 2013 Luis I. Terrazas et al. All rights reserved. Toxocara Seroprevalence in Patients with Idiopathic Parkinson's Disease: Chance Association or Coincidence? Wed, 26 Dec 2012 13:14:45 +0000 Most cases of idiopathic Parkinson disease (IPD) are believed to be due to a combination of genetic and environmental factors. The purpose of this study is to investigate the relationship between toxocariasis and Parkinson disease (PD). Patients were selected from people who were admitted to the Movement Disorders Branch, Neurology Department of Elazığ University Faculty of Medicine Elazığ, Turkey. We studied specific IgG antibodies against Toxocara canis (T. canis) in 50 patients with idiopathic Parkinson and 50 healthy volunteers. We investigated the clinical history of three patients infected with T. canis. We also studied specific IgG antibodies against Toxoplasma gondii in these groups. Antibodies anti-Toxocara canis were found in 3 idiopathic PD (6%) () and antibody titer was not found in control. A patient had history of the presence of dog in current dog ownership. We did not detect any statistically significant association between T. canis and IPD. But, we believe that further comprehensive studies are required for understanding whether there is a causal relation between toxocariasis and PD. We didn’t find possible association between Toxoplasma gondii and IPD (). Tuncay Çelik, Yüksel Kaplan, Eser Ataş, Derya Öztuna, and Said Berilgen Copyright © 2013 Tuncay Çelik et al. All rights reserved. Treg Cells Induced by rSSP4 Derived from T. cruzi Amastigotes Increase Parasitemia in an Experimental Chagas Disease Model Wed, 26 Dec 2012 12:01:45 +0000 Currently, there is a considerable controversy over the participation of Treg cells during Trypanosoma cruzi infection, the main point being whether these cells play a negative or a positive role. In this work, we found that the adoptive transfer of CD4+CD25+FOXP3+ T cells from rSSP4- (a recombinant Trypanosoma cruzi amastigote derived protein, previously shown to have immunomodulatory properties on macrophages) immunized BALB/c donors into syngenic recipients simultaneously with T. cruzi challenge reduces cardiac inflammation and prolongs hosts’ survival but increases blood parasitemia and parasite loads in the heart. These CD4+CD25+FOXP3+ Treg cells from immunized mice have a relatively TGF-β-dependent suppressive activity on CD4+ T cells. Therefore, regulatory CD4+CD25+ T cells play a positive role in the development of acute T. cruzi infection by inducing immunosuppressive activity that controls early cardiac inflammation during acute Chagas disease, prolonging survival, but at the same time promoting parasite growth. Y. Flores-García, J. L. Rosales-Encina, V. H. Rosales-García, A. R. Satoskar, and P. Talamás-Rohana Copyright © 2013 Y. Flores-García et al. All rights reserved. The Importance of the Nurse Cells and Regulatory Cells in the Control of T Lymphocyte Responses Wed, 26 Dec 2012 09:14:16 +0000 T lymphocytes from the immune system are bone marrow-derived cells whose development and activities are carefully supervised by two sets of accessory cells. In the thymus, the immature young T lymphocytes are engulfed by epithelial “nurse cells” and retained in vacuoles, where most of them (95%) are negatively selected and removed when they have an incomplete development or express high affinity autoreactive receptors. The mature T lymphocytes that survive to this selection process leave the thymus and are controlled in the periphery by another subpopulation of accessory cells called “regulatory cells,” which reduce any excessive immune response and the risk of collateral injuries to healthy tissues. By different times and procedures, nurse cells and regulatory cells control both the development and the functions of T lymphocyte subpopulations. Disorders in the T lymphocytes development and migration have been observed in some parasitic diseases, which disrupt the thymic microenvironment of nurse cells. In other cases, parasites stimulate rather than depress the functions of regulatory T cells decreasing T-mediated host damages. This paper is a short review regarding some features of these accessory cells and their main interactions with T immature and mature lymphocytes. The modulatory role that neurotransmitters and hormones play in these interactions is also revised. María Guadalupe Reyes García and Fernando García Tamayo Copyright © 2013 María Guadalupe Reyes García and Fernando García Tamayo. All rights reserved. A New Parasiticidal Compound in T. solium Cysticercosis Thu, 20 Dec 2012 17:31:34 +0000 The effect of 16α-bromoepiandrosterone (EpiBr), a dehydroepiandrosterone (DHEA) analogue, was tested on the cysticerci of Taenia solium, both in vitro and in vivo. In vitro treatment of T. solium cultures with EpiBr reduced scolex evagination, growth, motility, and viability in dose- and time-dependent fashions. Administration of EpiBr prior to infection with T. solium cysticerci in hamsters reduced the number and size of developed taenias in the intestine, compared with controls. These effects were associated to an increase in splenocyte proliferation in infected hamsters. These results leave open the possibility of assessing the potential of this hormonal analogue as a possible antiparasite drug, particularly in cysticercosis and taeniosis. Romel Hernández-Bello, Galileo Escobedo, Julio Cesar Carrero, Claudia Cervantes-Rebolledo, Charles Dowding, James Frincke, Chris Reading, and Jorge Morales-Montor Copyright © 2013 Romel Hernández-Bello et al. All rights reserved. Immune Responses Associated with Resistance to Haemonchosis in Sheep Thu, 20 Dec 2012 15:21:24 +0000 This paper examines the known immunological and genetic factors associated with sheep resistance to infection by Haemonchus contortus. Such resistance is an inheritable genetic trait (, 0.22–0.63) associated with certain sheep breeds. Resistant sheep do not completely reject the disease; they only harbor fewer parasites than susceptible sheep and therefore have a lower fecal egg count. Protective immune response to haemonchosis is an expression of genetic resistance. Genes associated with resistance and susceptibility are described. Genetically resistant sheep have nonspecific mechanisms that block the initial colonization by Haemonchus contortus larvae. These sheep also have an efficacious Th2 type response (e.g., increases in blood and tissue eosinophils, specific IgE class antibodies, mast cells, IL-5, IL-13, and TNFα) that protects them against the infection; in contrast, susceptible sheep do not efficiently establish this type of immune response. Finally, the main reported antigens of H. contortus were reviewed. Fernando Alba-Hurtado and Marco Antonio Muñoz-Guzmán Copyright © 2013 Fernando Alba-Hurtado and Marco Antonio Muñoz-Guzmán. All rights reserved. Tamoxifen Treatment in Hamsters Induces Protection during Taeniosis by Taenia solium Thu, 20 Dec 2012 11:01:19 +0000 Human neurocysticercosis by Taenia solium is considered an emergent severe brain disorder in developing and developed countries. Discovery of new antiparasitic drugs has been recently aimed to restrain differentiation and establishment of the T. solium adult tapeworm, for being considered a central node in the disease propagation to both pigs and humans. Tamoxifen is an antiestrogenic drug with cysticidal action on Taenia crassiceps, a close relative of T. solium. Thus, we evaluated the effect of tamoxifen on the in vitro evagination and the in vivo establishment of T. solium. In vitro, tamoxifen inhibited evagination of T. solium cysticerci in a dose-time dependent manner. In vivo, administration of tamoxifen to hamsters decreased the intestinal establishment of the parasite by 70%, while recovered tapeworms showed an 80% reduction in length, appearing as scolices without strobilar development. Since tamoxifen did not show any significant effect on the proliferation of antigen-specific immune cells, intestinal inflammation, and expression of Th1/Th2 cytokines in spleen and duodenum, this drug could exert its antiparasite actions by having direct detrimental effects upon the adult tapeworm. These results demonstrate that tamoxifen exhibits a strong cysticidal and antitaeniasic effect on T. solium that should be further explored in humans and livestock. Galileo Escobedo, M. Isabel Palacios-Arreola, Alfonso Olivos, Lorena López-Griego, and Jorge Morales-Montor Copyright © 2013 Galileo Escobedo et al. All rights reserved. Protein Palmitoylation and Pathogenesis in Apicomplexan Parasites Wed, 03 Oct 2012 08:07:26 +0000 Apicomplexan parasites comprise a broad variety of protozoan parasites, including Toxoplasma gondii, Plasmodium, Eimeria, and Cryptosporidium species. Being intracellular parasites, the success in establishing pathogenesis relies in their ability to infect a host-cell and replicate within it. Protein palmitoylation is known to affect many aspects of cell biology. Furthermore, palmitoylation has recently been shown to affect important processes in T. gondii such as replication, invasion, and gliding. Thus, this paper focuses on the importance of protein palmitoylation in the pathogenesis of apicomplexan parasites. Maria Martha Corvi, Andres Mariano Alonso, and Marina Cecilia Caballero Copyright © 2012 Maria Martha Corvi et al. All rights reserved. Immunology and Cell Biology of Parasitic Diseases 2011 Sun, 03 Jun 2012 14:38:33 +0000 Luis I. Terrazas, Abhay R. Satoskar, and Jorge Morales-Montor Copyright © 2012 Luis I. Terrazas et al. All rights reserved. Erratum to “Flagellar Motility of Trypanosoma cruzi Epimastigotes” Wed, 30 May 2012 08:41:05 +0000 G. Ballesteros-Rodea, M. Santillán, S. Martínez-Calvillo, and R. Manning-Cela Copyright © 2012 G. Ballesteros-Rodea et al. All rights reserved. Comment on “Emerging Functions of Transcription Factors in Malaria Parasite” Sun, 20 May 2012 10:31:41 +0000 Heather J. Painter and Manuel Llinás Copyright © 2012 Heather J. Painter and Manuel Llinás. All rights reserved. EhADH112 Is a Bro1 Domain-Containing Protein Involved in the Entamoeba histolytica Multivesicular Bodies Pathway Tue, 28 Feb 2012 09:33:18 +0000 EhADH112 is an Entamoeba histolytica Bro1 domain-containing protein, structurally related to mammalian ALIX and yeast BRO1, both involved in the Endosomal Sorting Complexes Required for Transport (ESCRT)-mediated multivesicular bodies (MVB) biogenesis. Here, we investigated an alternative role for EhADH112 in the MVB protein trafficking pathway by overexpressing 166 amino acids of its N-terminal Bro1 domain in trophozoites. Trophozoites displayed diminished phagocytosis rates and accumulated exogenous Bro1 at cytoplasmic vesicles which aggregated into aberrant complexes at late stages of phagocytosis, probably preventing EhADH112 function. Additionally, the existence of a putative E. histolytica ESCRT-III subunit (EhVps32) presumably interacting with EhADH112, led us to perform pull-down experiments with GST-EhVps32 and [35S]-labeled EhADH112 or EhADH112 derivatives, confirming EhVps32 binding to EhADH112 through its Bro1 domain. Our overall results define EhADH112 as a novel member of ESCRT-accessory proteins transiently present at cellular surface and endosomal compartments, probably contributing to MVB formation during phagocytosis. Cecilia Bañuelos, Guillermina García-Rivera, Israel López-Reyes, Leobardo Mendoza, Arturo González-Robles, Silvia Herranz, Olivier Vincent, and Esther Orozco Copyright © 2012 Cecilia Bañuelos et al. All rights reserved. Are Basophils Important Mediators for Helminth-Induced Th2 Immune Responses? A Debate Mon, 20 Feb 2012 12:08:12 +0000 Helminth parasites induce Th2 immune responses. Immunological mechanisms leading to Th2 induction are mainly dependent on IL-4. However, early source of IL-4 has not been precisely identified. Noticeably, basophils seem to be important mediators for inducing and maintaining the Th2 response probably because they secrete IL-4 and exert functions similar to APCs. Nevertheless, recent experimental evidence points that DCs could be also significant participants during this process. The involvement of basophils during memory responses is also discussed. Sonia Leon-Cabrera and Ana Flisser Copyright © 2012 Sonia Leon-Cabrera and Ana Flisser. All rights reserved. Plasmodium Riboprotein PfP0 Induces a Deviant Humoral Immune Response in Balb/c Mice Tue, 17 Jan 2012 10:48:00 +0000 Passive immunization with antibodies to recombinant Plasmodium falciparum P0 riboprotein (rPfP0, 61–316 amino acids) provides protection against malaria. Carboxy-terminal 16 amino acids of the protein (PfP0C0) are conserved and show 69% identity to human and mouse P0. Antibodies to this domain are found in 10–15% of systemic lupus erythematosus patients. We probed the nature of humoral response to PfP0C0 by repeatedly immunizing mice with rPfP0. We failed to raise stable anti-PfP0C0 hybridomas from any of the 21 mice. The average serum anti-PfP0C0 titer remained low (5.1±1.3×104). Pathological changes were observed in the mice after seven boosts. Adsorption with dinitrophenyl hapten revealed that the anti-PfP0C0 response was largely polyreactive. This polyreactivity was distributed across all isotypes. Similar polyreactive responses to PfP0 and PfP0C0 were observed in sera from malaria patients. Our data suggests that PfP0 induces a deviant humoral response, and this may contribute to immune evasion mechanisms of the parasite. Sulabha Pathak, K. Rajeshwari, Swati Garg, Sudarsan Rajagopal, Kalpesh Patel, Bidyut Das, Sylviane Pied, Balachandran Ravindran, and Shobhona Sharma Copyright © 2012 Sulabha Pathak et al. All rights reserved. Proteomic Analysis of Trypanosoma cruzi Epimastigotes Subjected to Heat Shock Thu, 12 Jan 2012 12:11:27 +0000 Trypanosoma cruzi is exposed to sudden temperature changes during its life cycle. Adaptation to these variations is crucial for parasite survival, reproduction, and transmission. Some of these conditions may change the pattern of genetic expression of proteins involved in homeostasis in the course of stress treatment. In the present study, the proteome of T. cruzi epimastigotes subjected to heat shock and epimastigotes grow normally was compared by two-dimensional gel electrophoresis followed by mass spectrometry for protein identification. Twenty-four spots differing in abundance were identified. Of the twenty-four changed spots, nineteen showed a greater intensity and five a lower intensity relative to the control. Several functional categories of the identified proteins were determined: metabolism, cell defense, hypothetical proteins, protein fate, protein synthesis, cellular transport, and cell cycle. Proteins involved in the interaction with the cellular environment were also identified, and the implications of these changes are discussed. Deyanira Pérez-Morales, Humberto Lanz-Mendoza, Gerardo Hurtado, Rodrigo Martínez-Espinosa, and Bertha Espinoza Copyright © 2012 Deyanira Pérez-Morales et al. All rights reserved. Flagellar Motility of Trypanosoma cruzi Epimastigotes Wed, 11 Jan 2012 15:31:53 +0000 The hemoflagellate Trypanosoma cruzi is the causative agent of American trypanosomiasis. Despite the importance of motility in the parasite life cycle, little is known about T. cruzi motility, and there is no quantitative description of its flagellar beating. Using video microscopy and quantitative vectorial analysis of epimastigote trajectories, we find a forward parasite motility defined by tip-to-base symmetrical flagellar beats. This motion is occasionally interrupted by base-to-tip highly asymmetric beats, which represent the ciliary beat of trypanosomatid flagella. The switch between flagellar and ciliary beating facilitates the parasite's reorientation, which produces a large variability of movement and trajectories that results in different distance ranges traveled by the cells. An analysis of the distance, speed, and rotational angle indicates that epimastigote movement is not completely random, and the phenomenon is highly dependent on the parasite behavior and is characterized by directed and tumbling parasite motion as well as their combination, resulting in the alternation of rectilinear and intricate motility paths. G. Ballesteros-Rodea, M. Santillán, S. Martínez-Calvillo, and R. Manning-Cela Copyright © 2012 G. Ballesteros-Rodea et al. All rights reserved. Regulatory T Cells and Parasites Thu, 29 Dec 2011 17:54:26 +0000 Human host encounters a wide array of parasites; however, the crucial aspect is the failure of the host immune system to clear these parasites despite antigen recognition. In the recent past, a new immunological concept has emerged, which provides a framework to better understand several aspects of host susceptibility to parasitic infection. It is widely believed that parasites are able to modulate the magnitude of effector responses by inducing regulatory T cell (Tregs) population and several studies have investigated whether this cell population plays a role in balancing protective immunity and pathogenesis during parasite infection. This review discusses the several mechanism of Treg-mediated immunosuppression in the human host and focuses on the functional role of Tregs and regulatory gene polymorphisms in infectious diseases. TP. Velavan and Olusola Ojurongbe Copyright © 2011 TP. Velavan and Olusola Ojurongbe. All rights reserved. The Hamster Model for Identification of Specific Antigens of Taenia solium Tapeworms Thu, 29 Dec 2011 14:36:29 +0000 Humans acquire taeniasis by ingesting pork meat infected with Taenia solium cysticerci, which are the only definitive hosts of the adult stage (tapeworm) and responsible for transmitting the human and porcine cysticercosis. Hence, detection of human tapeworm carriers is a key element in the development of viable strategies to control the disease. This paper presents the identification of specific antigens using sera from hamsters infected with T. solium tapeworms analyzed by western blot assay with crude extracts (CEs) and excretion-secretion antigens (E/S Ag) obtained from T. solium cysticerci and tapeworms and extracts from other helminthes as controls. The hamster sera infected with T. solium tapeworms recognized specific bands of 72, 48, 36, and 24 kDa, in percentages of 81, 81, 90, and 88%, respectively, using the T. solium tapeworms E/S Ag. The antigens recognized by these hamster sera could be candidates to improve diagnosis of human T. solium taeniasis. Alicia Ochoa-Sánchez, Lucía Jiménez, and Abraham Landa Copyright © 2011 Alicia Ochoa-Sánchez et al. All rights reserved.