Journal of Pathogens The latest articles from Hindawi Publishing Corporation © 2014 , Hindawi Publishing Corporation . All rights reserved. Antimicrobial Susceptibility of Escherichia coli Isolated from Fresh-Marketed Nile Tilapia (Oreochromis niloticus) Mon, 07 Apr 2014 09:51:49 +0000 The contamination of seafood by bacteria of fecal origin, especially Escherichia coli, is a widely documented sanitary problem. The objective of the present study was to isolate E. coli strains from the gills, muscle, and body surface of farmed Nile tilapias (Oreochromis niloticus) fresh-marketed in supermarkets in Fortaleza (Ceará, Brazil), to determine their susceptibility to antibiotics of different families (amikacin, gentamicin, imipenem, cephalothin, cefotaxime, ciprofloxacin, aztreonam, ampicillin, nalidixic acid, tetracycline, and sulfametoxazol-trimetoprim), and to determine the nature of resistance by plasmid curing. Forty-four strains (body surface = 25, gills = 15, muscle = 4) were isolated, all of which were susceptible to amikacin, aztreonam, cefotaxime, ciprofloxacin, gentamicin, and imipenem. Gill and body surface samples yielded 11 isolates resistant to ampicillin, tetracycline, and sulfametoxazol-trimetoprim, 4 of which of plasmidial nature. The multiple antibiotic resistance index was higher for strains isolated from body surface than from gills. The overall high antibiotic susceptibility of E. coli strains isolated from fresh-marketed tilapia was satisfactory, although the occasional finding of plasmidial resistance points to the need for close microbiological surveillance of the farming, handling, and marketing conditions of aquaculture products. Rafael dos Santos Rocha, Lana Oliveira Leite, Oscarina Viana de Sousa, and Regine Helena Silva dos Fernandes Vieira Copyright © 2014 Rafael dos Santos Rocha et al. All rights reserved. Detection of Legionella spp. from Domestic Water in the Prefecture of Arta, Greece Wed, 12 Mar 2014 07:36:38 +0000 The aim of this research was the isolation of Legionella spp. from domestic water supply networks in the Prefecture of Arta. A total of 100 water samples, from 25 houses, were collected. Half of the samples concerned the cold water and half the hot water supply. Purpose was to detect colonization of the water networks with Legionella spp. >500 cfu/L by using the method of filtration (ISO 11731). Out of 100 samples, 6 samples from 3 houses were positive for Legionella spp. Legionella pneumophila serogroup 2–14 was isolated in 5 of 6 samples, whereas in the sixth sample Legionella anisa was identified. Only three of the samples had residual chloride over 0.2 mg/L, rate which is necessary for potable water, according to the Greek hygienic practice. Concerning the temperature of hot water, the mean temperature of the negative for Legionella samples was higher compared to the mean temperature of the positive for Legionella samples (49.9°C versus 45.5°C). It is estimated that there is risk of infection through the use of showers. The low concentration of chloride and the temperature, which was found within the limits favorable to developing Legionella spp. (20–45°C), provide fertile ground for proliferation of the bacteria. Dimitra Dimitriadi and Emmanuel Velonakis Copyright © 2014 Dimitra Dimitriadi and Emmanuel Velonakis. All rights reserved. Correlation between PFGE Groups and mrp/epf/sly Genotypes of Human Streptococcus suis Serotype 2 in Northern Thailand Thu, 06 Mar 2014 09:01:46 +0000 Streptococcus suis infection is a severe zoonotic disease commonly found in Northern Thailand where people often consume raw pork and/or pig’s blood. The most frequent clinical presentations are meningitis, sepsis, and endocarditis with higher rate of mortality and hearing loss sequelae. To clarify the correlation between pulsed-field gel electrophoresis (PFGE) groups and mrp/epf/sly genotypes of S. suis serotype 2, 62 patient and 4 healthy pig isolates from Northern Thailand were studied. By PFGE analysis, at 66% homology, most human isolates (69.4%) and 1 pig isolate were in group A, whereas 14.5% of human isolates and 3 out of 4 pig isolates were in group D. According to mrp/epf/sly genotypes, 80.6% of human isolates were identified in mrp+epf−sly− and only 12.9% were in mrp−epf−sly+ genotypes; in contrast, 1 and 3 pig isolates were detected in these two genotypes, respectively. Interestingly, all isolates of S. suis serotype 2 classified in PFGE groups A, B, and E were set in mrp+epf−sly− genotypes. These data show a close correlation between PFGE groups and mrp/epf/sly genotypes of human S. suis serotype 2. Prasit Tharavichitkul, Kanreuthai Wongsawan, Naoki Takenami, Sumalee Pruksakorn, Achara Fongcom, Marcelo Gottschalk, Banyong Khanthawa, Volaluk Supajatura, and Shinji Takai Copyright © 2014 Prasit Tharavichitkul et al. All rights reserved. Tailoring the Immune Response via Customization of Pathogen Gene Expression Tue, 25 Feb 2014 13:02:41 +0000 The majority of studies focused on the construction and reengineering of bacterial pathogens have mainly relied on the knocking out of virulence factors or deletion/mutation of amino acid residues to then observe the microbe’s phenotype and the resulting effect on the host immune response. These knockout bacterial strains have also been proposed as vaccines to combat bacterial disease. Theoretically, knockout strains would be unable to cause disease since their virulence factors have been removed, yet they could induce a protective memory response. While knockout strains have been valuable tools to discern the role of virulence factors in host immunity and bacterial pathogenesis, they have been unable to yield clinically relevant vaccines. The advent of synthetic biology and enhanced user-directed gene customization has altered this binary process of knockout, followed by observation. Recent studies have shown that a researcher can now tailor and customize a given microbe’s gene expression to produce a desired immune response. In this commentary, we highlight these studies as a new avenue for controlling the inflammatory response as well as vaccine development. Lisa M. Runco, Charles B. Stauft, and J. Robert Coleman Copyright © 2014 Lisa M. Runco et al. All rights reserved. High Prevalence of Helicobacter pylori hopQ II Genotype Isolated from Iranian Patients with Gastroduodenal Disorders Mon, 10 Feb 2014 07:13:53 +0000 Helicobacter pylori plays an important role in the pathogenesis of chronic gastritis, peptic ulceration, and noncardia gastric cancer. Several putative virulence factors for H. pylori have been identified including vacA, babA, and iceA. HopQ is one of the outer membrane proteins involved in bacterial adherence to gastric mucosa and has been suggested to also play a role in the virulence of H. pylori. Due to the substantial geographic differences in the prevalence of H. pylori virulence factors reported, the main purpose of the current study was to investigate the association between different H. pylori virulence hopQ alleles (types I and II) and patients with gastroduodenal disorders. The presence of H. pylori and hopQ alleles in gastric biopsy specimens was identified by specific PCR assays. H. pylori type II hopQ was found to be significantly associated with gastric cancer patients (odds ratio: 3.47, 95% CI: 1.56–5.89). Information about the prevalence of H. pylori hopQ type II can be used for determining the high-risk diseases type which is actually colonized by H. pylori hopQ type II positive strains. The presence of H. pylori hopQ type II should be investigated in different geographical regions as confirmatory findings may provide a definite biomarker attributed to the pathogenesis of certain severe digestive diseases. Amin Talebi Bezmin Abadi and Ashraf Mohabbati Mobarez Copyright © 2014 Amin Talebi Bezmin Abadi and Ashraf Mohabbati Mobarez. All rights reserved. Pathogenicity, Ovicidal Action, and Median Lethal Concentrations (LC50) of Entomopathogenic Fungi against Exotic Spiralling Whitefly, Aleurodicus dispersus Russell Mon, 23 Dec 2013 09:34:13 +0000 Biological control using entomopathogenic fungi could be a promising alternative to chemical control. Entomopathogenic fungi, Beauveria bassiana (Balsamo) Vuillemin, Metarhizium anisopliae (Metschnikoff) Sorokin, Lecanicillium lecanii (Zimmerm.) Zare and Gams, and Paecilomyces fumosoroseus (Wize) Brown and Smith, were tested for their pathogenicity, ovicidal effect, and median lethal concentrations (LC50) against exotic spiralling whitefly, Aleurodicus dispersus Russell. The applications were made at the rate of 2 × 109 conidia mL−1 for evaluating the pathogenicity and ovicidal effect of entomopathogenic fungi against A. dispersus. The results of pathogenicity test showed that P. fumosoroseus (P1 strain) was highly pathogenic to A. dispersus recording 100% mortality at 15 days after treatment (DAT). M. anisopliae (M2 strain) had more ovicidal effect causing 37.3% egg mortality at 8 DAT. However, L. lecanii (L1 strain) caused minimum egg hatchability (23.2%) at 10 DAT as compared to control (92.6%). The lowest LC50 produced by P. fumosoroseus (P1 strain) as 8.189 × 107 conidia mL−1 indicated higher virulence against A. dispersus. Hence, there is potential for use of entomopathogenic fungi in the field conditions as an alternate control method in combating the insect pests and other arthropod pests since they are considered natural mortality agents and are environmentally safe. Boopathi Thangavel, Karuppuchamy Palaniappan, Kalyanasundaram Manickavasagam Pillai, Mohankumar Subbarayalu, and Ravi Madhaiyan Copyright © 2013 Boopathi Thangavel et al. All rights reserved. Antibiotic Resistance of Salmonella spp. Isolated from Shrimp Farming Freshwater Environment in Northeast Region of Brazil Sun, 22 Dec 2013 14:22:14 +0000 This study investigated the presence and antibiotic resistance of Salmonella spp. in a shrimp farming environment in Northeast Region of Brazil. Samples of water and sediments from two farms rearing freshwater-acclimated Litopenaeus vannamei were examined for the presence of Salmonella. Afterwards, Salmonella isolates were serotyped, the antimicrobial resistance was determined by a disk diffusion method, and the plasmid curing was performed for resistant isolates. A total of 30 (16.12%) of the 186 isolates were confirmed to be Salmonella spp., belonging to five serovars: S. serovar Saintpaul, S. serovar Infantis, S. serovar Panama, S. serovar Madelia, and S. serovar Braenderup, along with 2 subspecies: S. enterica serovar houtenae and S. enterica serovar enterica. About twenty-three percent of the isolates were resistant to at least one antibiotic, and twenty percent were resistant to at least two antibiotics. Three strains isolated from water samples (pond and inlet canal) exhibited multiresistance to ampicillin, tetracycline, oxytetracycline, and nitrofurantoin. One of them had a plasmid with genes conferring resistance to nitrofurantoin and ampicillin. The incidence of bacteria pathogenic to humans in a shrimp farming environment, as well as their drug-resistance pattern revealed in this study, emphasizes the need for a more rigorous attention to this area. Fátima C. T. Carvalho, Oscarina V. Sousa, Edirsana M. R. Carvalho, Ernesto Hofer, and Regine H. S. F. Vieira Copyright © 2013 Fátima C. T. Carvalho et al. All rights reserved. Effect of Commercial Cyanobacteria Products on the Growth and Antagonistic Ability of Some Bioagents under Laboratory Conditions Thu, 07 Nov 2013 18:01:31 +0000 Evaluation of the efficacy of blue-green algal compounds against the growth of either pathogenic or antagonistic microorganisms as well as their effect on the antagonistic ability of bioagents was studied under in vitro conditions. The present study was undertaken to explore the inhibitory effect of commercial algal compounds, Weed-Max and Oligo-Mix, against some soil-borne pathogens. In growth medium supplemented with these algal compounds, the linear growth of pathogenic fungi decreased by increasing tested concentrations of the two algal compounds. Complete reduction in pathogenic fungal growth was observed at 2% of both Weed-Max and Oligo-Mix. Gradual significant reduction in the pathogenic fungal growth was caused by the two bioagents and by increasing the concentrations of algal compounds Weed-Max and Oligo-Mix. The present work showed that commercial algal compounds, Weed-Max and Oligo-Mix, have potential for the suppression of soil-borne fungi and enhance the antagonistic ability of fungal, bacterial, and yeast bio-agents. Nehal S. El-Mougy and Mokhtar M. Abdel-Kader Copyright © 2013 Nehal S. El-Mougy and Mokhtar M. Abdel-Kader. All rights reserved. Proof of Principle for a Real-Time Pathogen Isolation Media Diagnostic: The Use of Laser-Induced Breakdown Spectroscopy to Discriminate Bacterial Pathogens and Antimicrobial-Resistant Staphylococcus aureus Strains Grown on Blood Agar Tue, 10 Sep 2013 08:18:28 +0000 Laser-Induced Breakdown Spectroscopy (LIBS) is a rapid, in situ, diagnostic technique in which light emissions from a laser plasma formed on the sample are used for analysis allowing automated analysis results to be available in seconds to minutes. This speed of analysis coupled with little or no sample preparation makes LIBS an attractive detection tool. In this study, it is demonstrated that LIBS can be utilized to discriminate both the bacterial species and strains of bacterial colonies grown on blood agar. A discrimination algorithm was created based on multivariate regression analysis of spectral data. The algorithm was deployed on a simulated LIBS instrument system to demonstrate discrimination capability using 6 species. Genetically altered Staphylococcus aureus strains grown on BA, including isogenic sets that differed only by the acquisition of mutations that increase fusidic acid or vancomycin resistance, were also discriminated. The algorithm successfully identified all thirteen cultures used in this study in a time period of 2 minutes. This work provides proof of principle for a LIBS instrumentation system that could be developed for the rapid discrimination of bacterial species and strains demonstrating relatively minor genomic alterations using data collected directly from pathogen isolation media. Rosalie A. Multari, David A. Cremers, Melissa L. Bostian, Joanne M. Dupre, and John E. Gustafson Copyright © 2013 Rosalie A. Multari et al. All rights reserved. Evaluation of Central Venous Catheter Associated Blood Stream Infections: A Microbiological Observational Study Tue, 09 Jul 2013 09:05:36 +0000 There are substantial morbidity and mortality associated with vascular catheter use among crictically ill patients. The attributable mortality is 10% to 25% which is associated with bacteremia among those who are hospitalized. This study was undertaken to identify catheter related blood stream infections, to isolate pathogenic microorganisms present in intravascular catheter related local infections, exit site infections, and to determine the predisposing factors for the development of such infections and antibiotic sensitivity pattern of the isolated organisms in tertiary care hospital. Vinay Khanna, Chiranjay Mukhopadhayay, Vandana K. E., Murlidhar Verma, and Partha Dabke Copyright © 2013 Vinay Khanna et al. All rights reserved. Genotypic Characterization of Yersinia enterocolitica Biotype 4/O:3 Isolates from Pigs and Slaughterhouses Using SE-AFLP, ERIC-PCR, and PFGE Thu, 30 May 2013 14:36:50 +0000 Yersinia enterocolitica is a foodborne pathogen that causes illness in humans and animals. The biotype 4/O:3 has been commonly associated with yersiniosis and is characterized by the presence of chromosomal and extra-chromosomal virulence genes. Molecular typing methods have been successfully used to characterize Y. enterocolitica genetic heterogeneity and to study the epidemiology of the bacteria from different origins. In this study, 320 Y. enterocolitica biotype 4/O:3 isolates originating in pigs and slaughterhouses were characterized according to the virulence profile, and 61 isolates were typified through SE-AFLP, ERIC-PCR, and PFGE techniques. The majority of the isolates originated from pigs, and the predominant virulence profile was ail+ virF+ rfbC+ ystA+, representing 83.4% of the tested isolates. All of the Y. enterocolitica 4/O:3 isolates were positive for at least ystA gene. The SE-AFLP and ERIC-PCR patterns were highly homogeneous. The SE-AFLP was more discriminative than the ERIC-PCR and tended to cluster isolates according to the slaughterhouse. Despite the limited genetic diversity of Y. enterocolitica 4/O:3, PFGE was shown to be the most discriminative technique considering one band of difference. Fattening pigs proved to be an important reservoir of Y. enterocolitica biotype 4/O:3 carrying virulence genes. Renata Paixão, Luisa Zanolli Moreno, Débora Dirani Sena de Gobbi, Daniele Cristine Raimundo, Thais Sebastiana Porfida Ferreira, Maria Garcia Spindola, Ernesto Hofer, Cristhiane Moura Falavina dos Reis, Maria Helena Matté, and Andrea Micke Moreno Copyright © 2013 Renata Paixão et al. All rights reserved. Probiotics as Antiviral Agents in Shrimp Aquaculture Tue, 30 Apr 2013 09:17:44 +0000 Shrimp farming is an aquaculture business for the cultivation of marine shrimps or prawns for human consumption and is now considered as a major economic and food production sector as it is an increasingly important source of protein available for human consumption. Intensification of shrimp farming had led to the development of a number of diseases, which resulted in the excessive use of antimicrobial agents, which is finally responsible for many adverse effects. Currently, probiotics are chosen as the best alternatives to these antimicrobial agents and they act as natural immune enhancers, which provoke the disease resistance in shrimp farm. Viral diseases stand as the major constraint causing an enormous loss in the production in shrimp farms. Probiotics besides being beneficial bacteria also possess antiviral activity. Exploitation of these probiotics in treatment and prevention of viral diseases in shrimp aquaculture is a novel and efficient method. This review discusses the benefits of probiotics and their criteria for selection in shrimp aquaculture and their role in immune power enhancement towards viral diseases. Bestha Lakshmi, Buddolla Viswanath, and D. V. R. Sai Gopal Copyright © 2013 Bestha Lakshmi et al. All rights reserved. Seroepidemiology of Helicobacter pylori Infection in Tepehuanos Aged 15 Years and Older in Durango, Mexico Wed, 20 Mar 2013 09:01:44 +0000 This study aimed to determine the seroepidemiology of Helicobacter pylori infection in Tepehuanos (an indigenous ethnic group living in rural Mexico). The prevalence of anti-Helicobacter pylori IgG antibodies was examined in 156 Tepehuanos in Durango State, Mexico, using an enzyme-linked immunoassay. In addition, sociodemographic, clinical, and behavioral characteristics of Tepehuanos associated with seropositivity were investigated. In total, 103 (66%) of the 156 participants (mean age years) had Helicobacter pylori IgG antibodies. Fifty-four (52.4%) of the 103 seropositive individuals had Helicobacter pylori IgG antibody levels higher than 100 U/mL. Males and females had comparable seroprevalence of Helicobacter pylori infection and Helicobacter pylori IgG antibody levels. The seroprevalence was significantly higher in women with pregnancies than those without this obstetric characteristic. Logistic regression showed that Helicobacter pylori infection was positively associated with low education (OR = 3.37; 95% CI: 1.13–10.00; ) and laborer occupation (OR = 2.71; 95% CI: 1.14–6.42; ). This is the first report of seroprevalence and contributing factors for Helicobacter pylori infection in Tepehuanos and of the association of Helicobacter pylori infection with laborer occupation. Results warrants further research. Cosme Alvarado-Esquivel Copyright © 2013 Cosme Alvarado-Esquivel. All rights reserved. Toxoplasmosis and Polygenic Disease Susceptibility Genes: Extensive Toxoplasma gondii Host/Pathogen Interactome Enrichment in Nine Psychiatric or Neurological Disorders Mon, 04 Mar 2013 15:00:02 +0000 Toxoplasma gondii is not only implicated in schizophrenia and related disorders, but also in Alzheimer's or Parkinson's disease, cancer, cardiac myopathies, and autoimmune disorders. During its life cycle, the pathogen interacts with ~3000 host genes or proteins. Susceptibility genes for multiple sclerosis, Alzheimer's disease, schizophrenia, bipolar disorder, depression, childhood obesity, Parkinson's disease, attention deficit hyperactivity disorder (multiple sclerosis), and autism (), but not anorexia or chronic fatigue are highly enriched in the human arm of this interactome and 18 (ADHD) to 33% (MS) of the susceptibility genes relate to it. The signalling pathways involved in the susceptibility gene/interactome overlaps are relatively specific and relevant to each disease suggesting a means whereby susceptibility genes could orient the attentions of a single pathogen towards disruption of the specific pathways that together contribute (positively or negatively) to the endophenotypes of different diseases. Conditional protein knockdown, orchestrated by T. gondii proteins or antibodies binding to those of the host (pathogen derived autoimmunity) and metabolite exchange, may contribute to this disruption. Susceptibility genes may thus be related to the causes and influencers of disease, rather than (and as well as) to the disease itself. C. J. Carter Copyright © 2013 C. J. Carter. All rights reserved. Experimental Airborne Transmission of Porcine Postweaning Multisystemic Wasting Syndrome Thu, 07 Feb 2013 10:23:19 +0000 The objective of these studies was to investigate if porcine postweaning multisystemic wasting syndrome (PMWS) could be induced in healthy pigs following contact with air from pigs with clinical signs of PMWS. The pigs were housed in different units. Either 31 (study I) or 25 (study II) pigs with clinical symptoms of PMWS from a PMWS-affected herd and 25 healthy pigs from a PMWS-free, but PCV2-positive, herd were housed in unit A. Fifty pigs from a PMWS-free herd were housed in unit B, which were connected by pipes to unit A. In unit C, 30 pigs from a PMWS-free herd were housed as controls. In study II, the pigs in units A and B from the PMWS-free herd developed clinical signs of PMWS 2-3 weeks after arrival. PMWS was confirmed at necropsy and the diseased pigs had increased PCV2 load and increased antibody titers against PCV2 in serum that coincided with the development of clinical signs typical of PMWS. Sequence analysis revealed that the PCV2 isolate belonged to genotype 2b. In conclusion, the present study showed that PMWS can be induced in pigs from a PMWS-free herd by airborne contact with pigs from a PMWS-affected herd. C. S. Kristensen, C. K. Hjulsager, K. Vestergaard, K. Dupont, V. Bille-Hansen, C. Enøe, S. E. Jorsal, P. Bækbo, and L. E. Larsen Copyright © 2013 C. S. Kristensen et al. All rights reserved. Monitoring of Active Human Herpes Virus 6 Infection in Iranian Patients with Different Subtypes of Multiple Sclerosis Tue, 22 Jan 2013 13:13:19 +0000 Background. Recently, it has been suggested that human herpes virus 6 (HHV6) may play a role in the pathogenesis of multiple sclerosis (MS). Our purpose is to determine the incidence of reactivated HHV6 in MS patients. Methods. Viral sequence analyzed by qPCR in the peripheral blood mononuclear cells (PBMCs), serum, and saliva samples of different subtypes of MS patients () and healthy controls (). HHV6 IgG and IgM antibody levels measured by ELISA technique in the plasma samples of both groups. Likewise, cerebrospinal fluid (CSF) samples of some MS patients () were analyzed for viral sequence. Results. Results demonstrate increased levels of anti-HHV6-IgG (78.2% versus 76.4% in controls; ), and IgM (34.6% versus 6.5% in controls; ) in MS patients. Furthermore, RRMS and SPMS patients showed relatively higher anti-HHV6 IgG and IgM compared to PPMS (). Moreover, load of cell-free viral DNA was higher in RRMS and SPMS patients and detected in 60.2% (47/78) of MS patients, compared with 14.6% (18/123) of healthy controls (). Moreover, load of cell-free viral DNA was higher in RRMS and SPMS patients and detected in 60.2% (47/78) of MS patients, compared with 14.6% (18/123) of healthy controls (). Conclusions. The results extend the observation of an increased frequency of systemic reactivated HHV6 infection in MS patients with developed stages of disease. Nourollah Ramroodi, Nima Sanadgol, Zohre Ganjali, Abbas Ali Niazi, Vida Sarabandi, and Ali Moghtaderi Copyright © 2013 Nourollah Ramroodi et al. All rights reserved. Aerobiology and Its Role in the Transmission of Infectious Diseases Sun, 13 Jan 2013 09:10:16 +0000 Aerobiology plays a fundamental role in the transmission of infectious diseases. As infectious disease and infection control practitioners continue employing contemporary techniques (e.g., computational fluid dynamics to study particle flow, polymerase chain reaction methodologies to quantify particle concentrations in various settings, and epidemiology to track the spread of disease), the central variables affecting the airborne transmission of pathogens are becoming better known. This paper reviews many of these aerobiological variables (e.g., particle size, particle type, the duration that particles can remain airborne, the distance that particles can travel, and meteorological and environmental factors), as well as the common origins of these infectious particles. We then review several real-world settings with known difficulties controlling the airborne transmission of infectious particles (e.g., office buildings, healthcare facilities, and commercial airplanes), while detailing the respective measures each of these industries is undertaking in its effort to ameliorate the transmission of airborne infectious diseases. Aaron Fernstrom and Michael Goldblatt Copyright © 2013 Aaron Fernstrom and Michael Goldblatt. All rights reserved. Clinical and Microbiological Profile of HIV/AIDS Cases with Diarrhea in North India Thu, 27 Dec 2012 18:10:01 +0000 Intestinal infections are a significant cause of morbidity and mortality in people living with HIV/AIDS (PLWHA) especially in developing countries. The present study was conducted to assess the clinical and microbiological spectrum in HIV/AIDS cases with diarrhea and to correlate the occurrence of such pathogens with stool characters, HIV seropositivity status, and CD4 counts. Stools from 154 HIV seropositive subjects and 50 HIV negative controls were examined by direct microscopy, fecal cultures, and serological tests (Clostridium difficile Toxin A, Cryptosporidium antigen, and Entamoeba histolytica antigen ELISA). CD4 T cell enumeration was done using FACS count (Becton Dickinson). The study showed a male preponderance (112 males and 42 females). Weakness, abdominal pain, and anorexia were the most common symptoms. Coccidian parasites were the most common cause of diarrhea in HIV seropositive cases. C. parvum was seen in 60.42% while Isospora belli in 9.03%. Amongst the bacterial pathogens C. difficile was detected in 18.06%, diarrheagenic Escherichia coli in 11.11%, and Shigella spp. in 2.78%. Pathogen isolation rates were more in HIV seropositive cases and subjects with low CD4 T lymphocyte counts. Regular monitoring of CD4 T lymphocyte counts and screening for enteric pathogens will help improve the quality of life for PLWHA. Arun Kumar Jha, Beena Uppal, Sanjim Chadha, Preena Bhalla, Roumi Ghosh, Prabhav Aggarwal, and Richa Dewan Copyright © 2012 Arun Kumar Jha et al. All rights reserved. Design and Construction of a Single-Tube, LATE-PCR, Multiplex Endpoint Assay with Lights-On/Lights-Off Probes for the Detection of Pathogens Associated with Sepsis Wed, 26 Dec 2012 14:34:45 +0000 Aims. The goal of this study was to construct a single tube molecular diagnostic multiplex assay for the detection of microbial pathogens commonly associated with septicemia, using LATE-PCR and Lights-On/Lights-Off probe technology. Methods and Results. The assay described here identified pathogens associated with sepsis by amplification and analysis of the 16S ribosomal DNA gene sequence for bacteria and specific gene sequences for fungi. A sequence from an unidentified gene in Lactococcus lactis subsp. cremoris served as a positive control for assay function. LATE-PCR was used to generate single-stranded amplicons that were then analyzed at endpoint over a wide temperature range in a specific fluorescent color. Each bacterial target was identified by its pattern of hybridization to Lights-On/Lights-Off probes derived from molecular beacons. Complex mixtures of targets were also detected. Conclusions. All microbial targets were identified in samples containing low starting copy numbers of pathogen genomic DNA, both as individual targets and in complex mixtures. Significance and Impact of the Study. This assay uses new technology to achieve an advance in the field of molecular diagnostics: a single-tube multiplex assay for identification of pathogens commonly associated with sepsis. Rachel K. Carver-Brown, Arthur H. Reis Jr., Lisa M. Rice, John W. Czajka, and Lawrence J. Wangh Copyright © 2012 Rachel K. Carver-Brown et al. All rights reserved. Identification at Biovar Level of Brucella Isolates Causing Abortion in Small Ruminants of Iran Tue, 27 Nov 2012 16:17:22 +0000 To determine the most prevalent biovar responsible for brucellosis in sheep and goat populations of Iran, a cross-sectional study was carried out over 2 years in six provinces selected based on geography and disease prevalence. Specimens obtained from referred aborted sheep and goat fetuses were cultured on Brucella selective media for microbiological isolation. Brucellae were isolated from 265 fetuses and examined for biovar identification using standard microbiological methods. Results showed that 246 isolates (92.8%) were B. melitensis biovar 1, 18 isolates (6.8%) were B. melitensis biovar 2, and, interestingly, one isolate (0.4%) obtained from Mazandaran province was B. abortus biovar 3. In this study, B. melitensis biovar 3 was isolated in none of the selected provinces, and all isolates from 3 provinces (i.e., Chehar-mahal Bakhtiari, Markazi, and Ilam) were identified only as B. melitensis biovar 1. In conclusion, we found that B. melitensis biovar 1 remains the most prevalent cause of small ruminant brucellosis in various provinces of Iran. Ali Mohammad Behroozikhah, Ramin Bagheri Nejad, Karim Amiri, and Ali Reza Bahonar Copyright © 2012 Ali Mohammad Behroozikhah et al. All rights reserved. Lipid Droplets and Mycobacterium leprae Infection Mon, 12 Nov 2012 15:10:19 +0000 Leprosy is a chronic infectious disease and is a major source of morbidity in developing countries. Leprosy is caused by the obligate intracellular bacterium Mycobacterium leprae, which infects as primary target Schwann cells. Lepromatous leprosy exhibits multiple lesions of the skin, eyes, nerves, and lymph nodes. The sites of infection are characterized by the presence of foamy macrophages, fully packed with lipid droplets (LDs), which are induced by M. leprae. In the last years, it has become evident that M. tuberculosis imports lipids from foamy macrophages and is dependent on fatty acids for growth in infected macrophages. M. leprae seems to have similar mechanisms for scavenging lipids from the host. But due to the inability to culture M. leprae on laboratory media, research progresses only slowly. However, in the last years, substantial progress has been made in the field of lipid metabolism in M. leprae. Herein, we will present and summarize the lipid droplets formation and the metabolism of lipids during M. leprae infection. Ayssar A. Elamin, Matthias Stehr, and Mahavir Singh Copyright © 2012 Ayssar A. Elamin et al. All rights reserved. A New Generation Microarray for the Simultaneous Detection and Identification of Yersinia pestis and Bacillus anthracis in Food Thu, 18 Oct 2012 08:53:37 +0000 The use of microarrays as a multiple analytic system has generated increased interest and provided a powerful analytical tool for the simultaneous detection of pathogens in a single experiment. A wide array of applications for this technology has been reported. A low density oligonucleotide microarray was generated from the genetic sequences of Y. pestis and B. anthracis and used to fabricate a microarray chip. The new generation chip, consisting of 2,240 spots in 4 quadrants with the capability of stripping/rehybridization, was designated as “Y-PESTIS/B-ANTHRACIS 4x2K Array.” The chip was tested for specificity using DNA from a panel of bacteria that may be potentially present in food. In all, 37 unique Y. pestis-specific and 83 B. anthracis-specific probes were identified. The microarray assay distinguished Y. pestis and B. anthracis from the other bacterial species tested and correctly identified the Y. pestis-specific oligonucleotide probes using DNA extracted from experimentally inoculated milk samples. Using a whole genome amplification method, the assay was able to detect as low as 1 ng genomic DNA as the start sample. The results suggest that oligonucleotide microarray can specifically detect and identify Y. pestis and B. anthracis and may be a potentially useful diagnostic tool for detecting and confirming the organisms in food during a bioterrorism event. Noriko Goji, Trevor MacMillan, and Kingsley Kwaku Amoako Copyright © 2012 Crown. All rights reserved. Human Sperm Interaction with Staphylococcus aureus: A Molecular Approach Mon, 15 Oct 2012 09:17:46 +0000 Sperm immobilization factor (SIF) causing 100% immobilization of spermatozoa isolated from Staphylococcus aureus when characterized using LC-MS (Liquid chromatography-mass spectrometry) showed that this 20 kDa protein had peptide sequence similarity with hsp-70 protein. It was found to completely (100%) inhibit Mg++ ATPase activity of spermatozoa at concentration of 100 μg mL−1. Sperm samples treated with SIF also showed reduction in calcium ionophore-induced acrosome reaction as compared to control samples (treated with calcium ionophore alone). Binding studies of FITC labelled SIF with spermatozoa using fluorescent microscopy showed binding of SIF to the surface of spermatozoa indicating the presence of SIF binding receptor. The receptor was extracted by 3M NaCl and purified by gel permeation chromatography. Characterization of the receptor by MALDI-TOF (Matrix-assisted laser desorption ionization-time of flight) indicated that the receptor shared sequence similarity with MHC class II antigen. A calorimetric study showed that the receptor moiety on spermatozoa was specific for the purified ligand as binding of the receptor to ligand was enthalpically (−11.9 kJ mole−1) as well as entropically (21.53 J mole−1 K−1) favored resulting in the Gibb's free energy of −18.57 kJ mole−1. Sonia Gupta and Vijay Prabha Copyright © 2012 Sonia Gupta and Vijay Prabha. All rights reserved. Understanding Viral Transmission Behavior via Protein Intrinsic Disorder Prediction: Coronaviruses Sun, 14 Oct 2012 08:32:24 +0000 Besides being a common threat to farm animals and poultry, coronavirus (CoV) was responsible for the human severe acute respiratory syndrome (SARS) epidemic in 2002–4. However, many aspects of CoV behavior, including modes of its transmission, are yet to be fully understood. We show that the amount and the peculiarities of distribution of the protein intrinsic disorder in the viral shell can be used for the efficient analysis of the behavior and transmission modes of CoV. The proposed model allows categorization of the various CoVs by the peculiarities of disorder distribution in their membrane (M) and nucleocapsid (N). This categorization enables quick identification of viruses with similar behaviors in transmission, regardless of genetic proximity. Based on this analysis, an empirical model for predicting the viral transmission behavior is developed. This model is able to explain some behavioral aspects of important coronaviruses that previously were not fully understood. The new predictor can be a useful tool for better epidemiological, clinical, and structural understanding of behavior of both newly emerging viruses and viruses that have been known for a long time. A potentially new vaccine strategy could involve searches for viral strains that are characterized by the evolutionary misfit between the peculiarities of the disorder distribution in their shells and their behavior. Gerard Kian-Meng Goh, A. Keith Dunker, and Vladimir N. Uversky Copyright © 2012 Gerard Kian-Meng Goh et al. All rights reserved. Rapid Detection and Identification of Yersinia pestis from Food Using Immunomagnetic Separation and Pyrosequencing Wed, 03 Oct 2012 11:39:35 +0000 Interest has recently been renewed in the possible use of Y. pestis, the causative agent of plague, as a biological weapon by terrorists. The vulnerability of food to intentional contamination coupled with reports of humans having acquired plague through eating infected animals that were not adequately cooked or handling of meat from infected animals makes the possible use of Y. pestis in a foodborne bioterrorism attack a reality. Rapid, efficient food sample preparation and detection systems that will help overcome the problem associated with the complexity of the different matrices and also remove any ambiguity in results will enable rapid informed decisions to be made regarding contamination of food with biothreat agents. We have developed a rapid detection assay that combines the use of immunomagnetic separation and pyrosequencing in generating results for the unambiguous identification of Y. pestis from milk (0.9 CFU/mL), bagged salad (1.6 CFU/g), and processed meat (10 CFU/g). The low detection limits demonstrated in this assay provide a novel tool for the rapid detection and confirmation of Y. pestis in food without the need for enrichment. The combined use of the iCropTheBug system and pyrosequencing for efficient capture and detection of Y. pestis is novel and has potential applications in food biodefence. Kingsley K. Amoako, Michael J. Shields, Noriko Goji, Chantal Paquet, Matthew C. Thomas, Timothy W. Janzen, Cesar I. Bin Kingombe, Arnold J. Kell, and Kristen R. Hahn Copyright © 2012 Crown. All rights reserved. Bacteria Isolations from Broiler and Layer Chicks in Zambia Mon, 03 Sep 2012 13:40:35 +0000 Chick mortality (CM) is one of the major constraints to the expansion of the poultry industry in Zambia. Of the 2,829 avian disease cases submitted to the national diagnostic laboratory based at the Central Veterinary Research Institute in Lusaka between 1995 and 2007, 34.39% (973/2,829) were from CM cases. The disease accounted for 40.2% (218,787/544,903) mortality in the affected flocks with 89.6% (196,112/218,787) of the affected birds dying within seven days. Major bacteria species involved were Escherichia coli, Salmonella gallinarum, and Proteus species being isolated from 84.58%, 46.15%, and 26.93% of the reported CM cases (𝑛=973), respectively. Detection of Salmonella typhimurium, Salmonella enteritidis, and Salmonella dublin indicates that poultry has the potential of transmitting zoonotic pathogenic bacteria to humans. The proportion of Salmonella gallinarum reactors in the adult breeding stock was generally low (<0.5%) throughout the study period although its prevalence in CM cases was correlated (𝑟=0.68, 𝑃<0.011) with seroprevalence of the same pathogen in the adult breeding stock. Given that the disease accounts for a large proportion of the avian diseases in Zambia as shown in the present study (34.39%, 𝑛=2,829), it is imperative that an effective disease control strategy aimed at reducing its occurrence should be developed. Hetron Mweemba Munang’andu, Swithine Hameenda Kabilika, Oliver Chibomba, Musso Munyeme, and Geoffrey Munkombwe Muuka Copyright © 2012 Hetron Mweemba Munang’andu et al. All rights reserved. Eleven-Year Distribution Pattern of Hepatitis C Virus in Southern Italy Thu, 09 Aug 2012 14:27:28 +0000 Analysis of the Hepatitis C Virus (HCV) genotype spread in a particular area has a crucial impact on public health. In this study, we update information on the distribution of HCV genotypes, by evaluating a hospital-based cohort of 2,153 chronic hepatitis C patients, collected prospectively among subjects attending University Hospital of Catanzaro, within an area of Southern Italy. We assessed the rates (%) of HCV genotypes during two consecutive periods, from 2001 to 2005 and from 2006 to 2011, according to age and gender. Considering overall observation time, subtype 1b was predominant followed by subtypes 2a/2c, genotype 3 and 4. Statistical evaluation of the age of HCV patients stratified by genotypes, revealed a slight but significant increase in the median age of 1b, 2a/2c and 3 HCV genotype-infected subjects, during the 2006–2011 period, whilst genotype 4 patients exhibited a decrease in the median age during the same period studied. Moreover genotype 4 increased between 2002 and 2003 as well as between 2010 and 2011. Due to the peculiar diagnostic/clinical/therapeutic features of HCV-4, our findings warrant a deeper investigation to better control infections caused by such genotype. Nadia Marascio, Giovanni Matera, Angela Quirino, Aida Giancotti, Giorgio S. Barreca, Angelo G. Lamberti, Benedetto Caroleo, Maria Carla Liberto, and Alfredo Focà Copyright © 2012 Nadia Marascio et al. All rights reserved. Detection and Quantification of Legionella pneumophila from Water Systems in Kuwait Residential Facilities Tue, 24 Jul 2012 08:05:33 +0000 The prevalence of Legionella pneumophilia in water systems of residential facilities in Kuwait was performed during the period from November 2007 to November 2011. A total of 204 water samples collected from faucets and showerheads in bathrooms (n = 82), taps in kitchens (n = 51), and water tanks (n = 71), from different locations of residential facilities in Kuwait were screened for Legionella pneumophila by the standard culture method and by real time polymerase chain reaction (RT-PCR). Out of the 204 samples, 89 (43.6%) samples were positive for Legionella spp., 48 (23.5%) samples were detected by the standard culture method, and 85 (41.7%) were detected by RT-PCR. Of the culture positive Legionella samples, counts ranged between 10 to 2250 CFU/L. Serological typing of 48 Legionella isolates revealed that 6 (12.5%) of these isolates belonged to Legionella pneumophila serogroup 1, 37 (77.1%) isolates to Legionella pneumophila serogroup 3, and 1 isolate each (2.1%) belonged to serogroups 4, 7, and 10. The minimum inhibitory concentration (MICs) of the 46 environmental L. pneumophila isolates against the 10 antimicrobials commonly used for Legionella infection treatments were determined. Rifampicin was found to be the most active against L. pneumophila serogroups isolates in vitro. Qadreyah A. Al-Matawah, Sameer F. Al-Zenki, Jafer A. Qasem, Tahani E. Al-Waalan, and Ahmed H. Ben Heji Copyright © 2012 Qadreyah A. Al-Matawah et al. All rights reserved. Yersiniosis and Food Safety Tue, 20 Mar 2012 16:21:32 +0000 Latiful Bari, Dike O. Ukuku, Kenji Isshiki, Ramesh C. Ray, and Didier Montet Copyright © 2012 Latiful Bari et al. All rights reserved. Averting Behavior Framework for Perceived Risk of Yersinia enterocolitica Infections Wed, 07 Mar 2012 08:38:23 +0000 The focus of this research is to present a theoretical model of averting actions that households take to avoid exposure to Yersinia enterocolitica in contaminated food. The cost of illness approach only takes into account the value of a cure, while the averting behavior approach can estimate the value of preventing the illness. The household, rather than the individual, is the unit of analysis in this model, where one household member is primarily responsible for procuring uncontaminated food for their family. Since children are particularly susceptible and live with parents who are primary decision makers for sustenance, the designated household head makes the choices that are investigated in this paper. This model uses constrained optimization to characterize activities that may offer protection from exposure to Yersinia enterocolitica contaminated food. A representative household decision maker is assumed to allocate family resources to maximize utility of an altruistic parent, an assumption used in most research involving economics of the family. Sonia N. Aziz and Khwaja M. S. Aziz Copyright © 2012 Sonia N. Aziz and Khwaja M. S. Aziz. All rights reserved.