Journal of Pathogens The latest articles from Hindawi © 2018 , Hindawi Limited . All rights reserved. Molecular Detection of Brucella spp. from Milk of Seronegative Cows from Some Selected Area in Bangladesh Sun, 14 Jan 2018 07:33:52 +0000 Brucellosis is endemic in Bangladesh both in humans and in animals. A number of reasons complicate the diagnosis, as bovine brucellosis can be diagnosed by various serological tests. But the tests have a limitation; when the organism remains intracellular, the disease goes into chronic stage and the antibody titres may decline. The present study was conducted for isolation and detection of Brucella spp. by polymerase chain reaction (PCR) from seronegative cows. A total of 360 dairy cows from three geographical regions were screened serologically by Rose Bengal Plate Test (RBPT) where 24 samples were serologically positive and the rest of the samples were serologically negative. Among the 24 seropositive individuals, 11 were culture positive and 6 were culture positive from serologically negative dairy cows. The overall seroprevalence of brucellosis in cattle was 6.6% and in disease condition a higher prevalence was recorded in abortion (28.07%) followed by infertility (13.33%). To confirm the Brucella spp. in seronegative dairy cattle, the isolates were extracted and PCR was conducted, which produced 905 bp amplicon size of 6 Brucella spp. from milk sample. So, for the detection or eradication of brucellosis, a bacteriological test and a PCR technique should be performed with the serological test of milk. Md. Sadequl Islam, Md. Ariful Islam, Mst. Minara Khatun, Sukumar Saha, Md. Samiul Basir, and Md- Mahmodul Hasan Copyright © 2018 Md. Sadequl Islam et al. All rights reserved. Epstein-Barr Virus as a Promising Immunotherapeutic Target for Nasopharyngeal Carcinoma Treatment Sun, 31 Dec 2017 11:20:12 +0000 Epstein-Barr virus (EBV) is a pathogen that infects more than 90% of global human population. EBV primarily targets B-lymphocytes and epithelial cells while some of them infect monocyte/macrophage, T-lymphocytes, and dendritic cells (DCs). EBV infection does not cause death by itself but the infection has been persistently associated with certain type of cancers such as nasopharyngeal carcinoma (NPC), Burkitt’s lymphoma (BL), and Hodgkin’s lymphoma (HL). Recent findings have shown promise on targeting EBV proteins for cancer therapy by immunotherapeutic approach. Some studies have also shown the success of adopting EBV-based therapeutic vaccines for the prevention of EBV-associated cancer particularly on NPC. In-depth investigations are in progress to refine the current therapeutic and vaccination strategies. In present review, we discuss the highly potential EBV targets for NPC immunotherapy and therapeutic vaccine development as well as addressing the underlying challenges in the process of bringing the therapy and vaccination from the bench to bedside. Sin-Yeang Teow, Hooi-Yeen Yap, and Suat-Cheng Peh Copyright © 2017 Sin-Yeang Teow et al. All rights reserved. Comparison of Algorithms for the Detection of Enteroviruses in Stool Specimens from Children Diagnosed with Acute Flaccid Paralysis Thu, 28 Dec 2017 00:00:00 +0000 This study was designed to compare both the cell culture dependent and independent enterovirus detection algorithms recommended by the WHO and assess how either might impact our perception of the diversity of enterovirus types present in a sample. Sixteen paired samples (16 isolates from RD cell culture and their corresponding stool suspension, i.e., 32 samples) from AFP cases in Nigeria were analyzed in this study. All the samples were subjected to RNA extraction, cDNA synthesis, the WHO recommended RT-snPCR, and its modification. Amplicons were sequenced and strains identified. Enterovirus diversity was the same between the isolates and fecal suspension for the control and five of the samples. It was, however, different for the remaining 10 (62.5%) samples. Nine (CV-B4, E6, E7, E13, E14, E19, E29, EV-B75, and EV-B77) and five (CV-A1, CV-A11, CV-A13, EV-C99, and PV2) EV-B and EV-C types, respectively, were detected. Particularly, E19 and EV-B75 were only recovered from the isolates while E14, EV-B77, CV-A11, and CV-A13 were only recovered from fecal suspension. Both the cell culture dependent and independent protocols bias our perception of the diversity of enterovirus types present in a sample. Hence, effort should be directed at harmonizing both for increased sensitivity. J. A. Adeniji, F. A. Ayeni, A. Ibrahim, K. A. Tijani, T. O. C. Faleye, and M. O. Adewumi Copyright © 2017 J. A. Adeniji et al. All rights reserved. Temporal Regulation of fim Genes in Uropathogenic Escherichia coli during Infection of the Murine Urinary Tract Wed, 27 Dec 2017 00:00:00 +0000 Uropathogenic Escherichia coli (UPEC) adhere to cells in the human urinary tract via type 1 pili that undergo phase variation where a 314-bp fimS DNA element flips between Phase-ON and Phase-OFF orientations through two site-specific recombinases, FimB and FimE. Three fim-lux operon transcriptional fusions were created and moved into the clinical UPEC isolate NU149 to determine their temporal regulation in UPEC growing in the urinary tract. Within murine urinary tracts, the UPEC strains demonstrated elevated transcription of fimA and fimB early in the infection, but lower transcription by the fifth day in murine kidneys. In contrast, fimE transcription was much lower than either fimA or fimB early, increased markedly at 24 h after inoculation, and then dropped five days after inoculation. Positioning of fimS was primarily in the Phase-ON position over the time span in UPEC infected bladders, whereas in UPEC infected murine kidneys the Phase-OFF orientation was favored by the fifth day after inoculation. Hemagglutination titers with guinea pig erythrocytes remained constant in UPEC growing in infected murine bladders but fell substantially in UPEC infected kidneys over time. Our results show temporal in vivo regulation of fim gene expression in different environmental niches when UPEC infects the murine urinary tract. William R. Schwan and Hua Ding Copyright © 2017 William R. Schwan and Hua Ding. All rights reserved. Analysis of Aminoglycoside Modifying Enzyme Genes Responsible for High-Level Aminoglycoside Resistance among Enterococcal Isolates Sun, 24 Dec 2017 08:58:20 +0000 Enzymatic modification results in high-level resistance to aminoglycoside (HLAR), which eliminates the synergistic bactericidal effect of combined exposure to a cell wall-active agent and an aminoglycoside. So aim of the study was to determine prevalence of HLAR enterococcal isolate and to study distribution of aminoglycoside modifying enzyme genes in them. A total of 100 nonrepeat isolates of enterococci from various clinical samples were analyzed. As per Clinical and Laboratory Standards Institute guidelines enterococci were screened for HLAR by Kirby-Bauer disc diffusion method. Minimum inhibitory concentration of all isolates for gentamicin and streptomycin was determined by E-test. Multiplex polymerase chain reaction (PCR) was carried out for HLAR enterococcal isolates to identify aminoglycoside modifying enzymes genes responsible for resistance. 60% isolates were found to be high-level gentamicin resistant (HLGR) whereas 45% isolates were found to be high-level streptomycin resistant (HLSR). By multiplex PCR 80% HLGR isolates carried bifunctional aminoglycoside modifying enzyme gene aac(6′)-Ie-aph(2′′)-Ia whereas 18 out of 45 high-level streptomycin resistant, that is, 40%, isolates carried aph(3′)-IIIa. However, aph(2′′)-Ib, aph(2′′)-Ic, aph(2′′)-Id, and ant(4′)-Ia genes which encode other aminoglycosides modifying enzymes were not detected. Bifunctional aminoglycoside modifying enzyme gene aac(6′)-Ie-aph(2′′)-Ia is the predominant gene responsible for HLAR. Vishal Shete, Naveen Grover, and Mahadevan Kumar Copyright © 2017 Vishal Shete et al. All rights reserved. Novel Genetic Variants of Hepatitis B Virus in Fulminant Hepatitis Tue, 19 Dec 2017 00:00:00 +0000 Fulminant hepatitis (FH) is a life-threatening liver disease characterised by intense immune attack and massive liver cell death. The common precore stop codon mutation of hepatitis B virus (HBV), A1896, is frequently associated with FH, but lacks specificity. This study attempts to uncover all possible viral nucleotides that are specifically associated with FH through a compiled sequence analysis of FH and non-FH cases from acute infection. We retrieved 67 FH and 280 acute non-FH cases of hepatitis B from GenBank and applied support vector machine (SVM) model to seek candidate nucleotides highly predictive of FH. Six best candidates with top predictive accuracy, 92.5%, were used to build a SVM model; they are C2129 (85.3%), T720 (83.0%), Y2131 (82.4%), T2013 (82.1%), K2048 (82.1%), and A2512 (82.1%). This model gave a high specificity (99.3%), positive predictive value (95.6%), and negative predictive value (92.1%), but only moderate sensitivity (64.2%). We successfully built a SVM model comprising six variants that are highly predictive and specific for FH: four in the core region and one each in the polymerase and the surface regions. These variants indicate that intracellular virion/core retention could play an important role in the progression to FH. Jack Bee Chook, Yun Fong Ngeow, Kok Keng Tee, Suat Cheng Peh, and Rosmawati Mohamed Copyright © 2017 Jack Bee Chook et al. All rights reserved. Acute Hepatitis E Virus Infection in Two Geographical Regions of Nigeria Wed, 13 Dec 2017 08:04:51 +0000 Hepatitis E virus (HEV) remains a major public health concern in resource limited regions of the world. Yet data reporting is suboptimal and surveillance system is inadequate. In Nigeria, there is dearth of information on prevalence of acute HEV infection. This study was therefore designed to describe acute HEV infection among antenatal clinic attendees and community dwellers from two geographical regions in Nigeria. Seven hundred and fifty plasma samples were tested for HEV IgM by Enzyme Linked Immunosorbent Assay (ELISA) technique. The tested samples were randomly selected from a pool of 1,115 blood specimens previously collected for viral hepatitis studies among selected populations (pregnant women, 272; Oyo community dwellers, 438; Anambra community dwellers, 405) between September 2012 and August 2013. One (0.4%) pregnant woman in her 3rd trimester had detectable HEV IgM, while community dwellers from the two study locations had zero prevalence rates of HEV IgM. Detection of HEV IgM in a pregnant woman, especially in her 3rd trimester, is of clinical and epidemiological significance. The need therefore exists for establishment of a robust HEV surveillance system in Nigeria and especially amidst the pregnant population in a bid to improve maternal and child health. I. M. Ifeorah, T. O. C. Faleye, A. S. Bakarey, M. O. Adewumi, A. Akere, E. C. Omoruyi, A. O. Ogunwale, and J. A. Adeniji Copyright © 2017 I. M. Ifeorah et al. All rights reserved. Evaluation of CAMP-Like Effect, Biofilm Formation, and Discrimination of Candida africana from Vaginal Candida albicans Species Sun, 26 Nov 2017 07:36:58 +0000 Candida africana as a species recovered from female genital specimens is highly close to C. albicans. The present study was conducted to discriminate C. africana from presumptive vaginal C. albicans strains by molecular assay and evaluate their hemolysin activity, biofilm formation, and cohemolytic effect (CAMP) with vaginal bacterial flora. A total of 110 stock vaginal C. albicans isolates were examined by HWP1 gene amplification. Hemolysin activity and the ability of biofilm formation were evaluated by blood plate assay and visual detection methods, respectively. Staphylococcus aureus, Staphylococcus epidermidis, and Streptococcus agalactiae were used to evaluate the CAMP-like effects in Sabouraud blood agar media. Based on the size of the amplicons (941 bp), all isolates were identified as C. albicans. All samples were able to produce beta-hemolysin. Moreover, 69 out of 110 of the isolates (62.7%) were biofilm-positive, 54 out of 110 Candida isolates (49%) demonstrated cohemolytic effects with S. agalactiae, and 48 out of 110 showed this effect with S. aureus (43.6%). All isolates were CAMP-negative with S. epidermidis. We detected all isolates as Candida albicans and almost half of the isolates were CAMP-positive with S. aureus and S. agalactiae, suggesting that these bacteria increase the pathogenicity of Candida in vaginal candidiasis. Keyvan Pakshir, Mahboubeh Bordbar, Kamiar Zomorodian, Hasti Nouraei, and Hossein Khodadadi Copyright © 2017 Keyvan Pakshir et al. All rights reserved. A Laboratory Assessment of Two Local Strains of the Beauveria bassiana (Bals.) Vuill. against the Tetranychus urticae (Acari: Tetranychidae) and Their Potential as a Mycopesticide Wed, 08 Nov 2017 00:00:00 +0000 This study was conducted to assess highly pathogenic Beauveria bassiana isolates to be used in biocontrol and to determine their potentials as mycopesticide. For this purpose, two B. bassiana isolates, which were locally isolated from T. urticae, were chosen. Firstly, three suspensions were investigated at the degree of humidity of 65 ± 5% and 100% RH. Secondly, these strains were selected according to their tendency to mass production, tolerance to UV radiation, and capability of producing spore at the different temperatures. Finally, identification of the selected isolate was performed by using ITS rDNA analysis. Both tested fungal isolates were pathogenic to the T. urticae. Mycelial growths of isolate AT076 at 20°C and 30°C were found to be greater than isolate AT007. It was observed that isolate AT076 had more spore production with 1.61 × 107 spore/disc at 30°C and 44.33% germination after UV radiation for 15 min. The numbers of spores per 5 mm disk area for isolates AT076 and AT007 were found to be 1.2 × 106 and 1.0 × 106. These results show that isolate AT076 was more virulent and more UV-tolerant and had higher tendency to mass production compared to isolate AT007 against T. urticae. As a result of this study, isolate AT076 can be used in the biocontrol as mycopesticide. Serkan Ortucu and Omer Faruk Algur Copyright © 2017 Serkan Ortucu and Omer Faruk Algur. All rights reserved. Molecular Epidemiology of Rifampicin Resistance in Mycobacterium tuberculosis Using the GeneXpert MTB/RIF Assay from a Rural Setting in India Thu, 26 Oct 2017 08:41:59 +0000 The Xpert MTB/RIF assay can detect mutations in rpoB gene that confer rifampicin resistance (RR) using five overlapping probes (A, B, C, D, and E). In this study, we described our experience with the Xpert assay in a rural setting in India. During the study period, 3250 samples were processed. The result was unsuccessful in 5.7% of cases. For extrapulmonary specimens, the risk of unsuccessful result was higher in tissue biopsy and stool samples. Among samples positive for Mycobacterium tuberculosis, rifampicin resistance was indeterminate in 1.2% of them. Our results and a review of the literature showed that the most frequent mutations conferring RR were located in the region of Probe E (63.6%; 95% confidence interval [CI] 56.26–70.94), followed by Probe B (15.02%; 95% CI 11.94–18.10), Probe D (13.35%; 95% CI 10.01–16.69), Probe A (4.73%; 95% CI 1.92–7.54), and Probe C (1.61%; 95% CI 0.67–2.54). Although the high cost of the cartridges precluded using the Xpert assay for routine diagnosis of tuberculosis, our results demonstrate that the assay can be used to diagnose RR-tuberculosis in rural areas with limited laboratory infrastructure and could be a convenient tool to investigate the molecular epidemiology of RR in resource-limited settings. Raghuprakash Reddy and Gerardo Alvarez-Uria Copyright © 2017 Raghuprakash Reddy and Gerardo Alvarez-Uria. All rights reserved. High Burden of Antimicrobial Resistance among Bacteria Causing Pyogenic Wound Infections at a Tertiary Care Hospital in Kathmandu, Nepal Mon, 28 Aug 2017 00:00:00 +0000 Pyogenic wound infections are one of the most common clinical entities caused and aggravated by the invasion of pathogenic organisms. Prompt and aggressive antimicrobial therapy is needed to reduce the burden and complications associated with these infections. In this study, we intended to investigate the common pathogens and their antimicrobial susceptibility patterns from the pyogenic wound infections at a tertiary care hospital in Kathmandu, Nepal. A laboratory based cross-sectional study was carried out among the pyogenic clinical specimens of the patients visiting Manmohan Memorial Teaching Hospital, Kathmandu, Nepal. Processing of clinical specimens and isolation and identification of bacterial pathogens were carried out using standard microbiological methods. Antimicrobial susceptibilities and resistant profiles were determined by following the standard guidelines of Clinical and Laboratory Standards Institute (CLSI). About 65% of the clinical specimens were positive for the bacterial growth and Gram positive bacteria (57.4%) were the leading pathogens among pyogenic wound infections. Staphylococcus aureus (412, 49.28%), Escherichia coli (136, 16.27%), Klebsiella spp. (88, 10.53%), and Pseudomonas spp. (44, 5.26%) were the common pathogens isolated. High level of drug resistance was observed among both Gram positive bacteria (51.9%) and Gram negative bacteria (48.7%). Gram positive isolates were resistant to ampicillin, ciprofloxacin, cotrimoxazole, erythromycin, and cloxacillin. Gram negative isolates were resistant to cephalosporins but were well susceptible to amikacin and imipenem. Pyogenic wound infections are common in our hospital and majority of them were associated with multidrug resistant bacteria. The detailed workup of the prevalent pathogens present in infected wounds and their resistance pattern is clearly pertinent to choosing the adequate treatment. Basista Prasad Rijal, Deepa Satyal, and Narayan Prasad Parajuli Copyright © 2017 Basista Prasad Rijal et al. All rights reserved. Histoplasmosis by Histoplasma capsulatum var. duboisii Observed at the Laboratory of Pathological Anatomy of Lomé in Togo Tue, 18 Jul 2017 00:00:00 +0000 Our study aimed to describe the epidemiological, clinical, and diagnostic aspects of African histoplasmosis in Togo through a descriptive and cross-sectional study on histological diagnosed African histoplasmosis in Pathology Department of Lomé from 2002 to 2016 (15 years). A total of 17 cases of African histoplasmosis were diagnosed. The sex ratio (M/F) was 1.8. The annual incidence was 1.1 cases. The mean age of the patients was 27.2 ± 0.4 years. All our patients were of social categories with a low socioeconomic level. HIV infection was known in 3 patients and one patient contracted tuberculosis. The clinical manifestations were cutaneous in 7 cases, cutaneous and mucous in 3 cases, cutaneous and lymph node in 3 cases, cutaneous and bone in 2 cases, and disseminated in 2 cases. The samples examined consisted of 14 cutaneous biopsies measuring 2-3 cm and 3 ganglionic biopsies each measuring 4 cm of major axis. Histologically, all cases were of chronic form made of granulomatous reaction with ovoid yeasts measuring between 1 and 2 microns. Despite the low frequency of this disease in our country, it should be kept constantly in mind before any granulomatous lesions, especially in the context of the HIV pandemic. Tchin Darré, Bayaki Saka, Abas Mouhari-Touré, Améyo Monique Dorkenoo, Koffi Amégbor, Vincent Palokinam Pitche, and Gado Napo-Koura Copyright © 2017 Tchin Darré et al. All rights reserved. Comparative Response of the Nigerian Indigenous and Broiler Chickens to a Field Caecal Isolate of Eimeria Oocysts Mon, 24 Apr 2017 05:59:15 +0000 Response of Nigerian indigenous (local) and broiler chickens to experimental Eimeria infections was investigated by measures of clinical signs, packed cell volume (PCV), body weights (BW), feed consumption, faecal oocyst counts (oocyst per gram), and microscopic intestinal lesions. Three-week-old chickens of each breed received single pulse infections with 2500, 5000, and 100.000 sporulated Eimeria oocysts. Infected birds were dull and passed bloody diarrhoea. OPG showed a dose related response but no significant difference between groups (). OPG was significantly higher in local chickens () and varied significantly with time (). PCV declined significantly in infected birds within breeds and groups (); however, the decline in PCV was significantly greater in broilers (). Both breeds had significant BW gains (). BW gain varied between groups being significantly higher in the uninfected control broilers than in the infected broilers (). Comparatively, broilers gained significantly more BW than their local counterparts (). Feed intake increased significantly with time () in both breeds. The Eimeria isolate was pathogenic to both breeds of chicken although clinical signs and lesions were more severe in indigenous chickens suggesting the breed’s more susceptibility. Lucas Atehmengo Ngongeh, Amaechi Onyeabor, Emeka Nzenwata, and Gurama Kansalem Samson Copyright © 2017 Lucas Atehmengo Ngongeh et al. All rights reserved. A Perspective of the Diagnosis and Management of Congenital Tuberculosis Thu, 24 Nov 2016 13:45:39 +0000 Tuberculosis continues to be a prevalent disease in the world and a global public health issue in many countries. The disease is more complicated in pregnant women because it imperils unborn offspring and results in congenital tuberculosis later if undiagnosed and untreated. Congenital tuberculosis is rare entity and an uncommon disease along with a high mortality rate. Congenital tuberculosis, a severe clinical type of tuberculosis caused by Mycobacterium tuberculosis, is a serious and fatal disease if left untreated. Our study emphasizes that it is necessary and mandatory to consider congenital tuberculosis in the differential diagnosis of neonatal or pulmonary infections in infants, essentially in countries where the incidence of tuberculosis is high burden. Mother to neonatal transmission of disease is well known via transplacental transmission through the umbilical vein to the fetus, through the ingestion of infected amniotic fluid. Early detection is challenging, because of the nonspecific nature of the signs and symptoms in tuberculosis during pregnancy and infancy. The degree of clinical suspicion is the essential component of diagnosis. Furthermore, it generally has a difficult treatment and it should not be delayed while waiting for diagnostic test results. Prompt identification and proper treatment regimens for congenital tuberculosis strongly relate with enhanced outcomes. Manou Irmina Saramba and Dongchi Zhao Copyright © 2016 Manou Irmina Saramba and Dongchi Zhao. All rights reserved. Etiology and Clinical Characteristics of Single and Multiple Respiratory Virus Infections Diagnosed in Croatian Children in Two Respiratory Seasons Tue, 30 Aug 2016 09:18:21 +0000 The aim of this study was to determine the causative agent of acute respiratory infection (ARI) in hospitalized children, as well as investigate the characteristics of ARIs with single and multiple virus detection in two respiratory seasons. In 2010 and 2015, nasopharyngeal and pharyngeal swabs from a total of 134 children, admitted to the hospital due to ARI, were tested using multiplex PCR. Viral etiology was established in 81.3% of the patients. Coinfection with two viruses was diagnosed in 27.6% of the patients, and concurrent detection of three or more viruses was diagnosed in 12.8% of the patients. The most commonly diagnosed virus in both seasons combined was respiratory syncytial virus (RSV) (28.6%), followed by parainfluenza viruses (PIVs) types 1–3 (18.4%), rhinovirus (HRV) (14.3%), human metapneumovirus (10.1%), adenovirus (AdV) (7.1%), influenza viruses types A and B (4.8%), and coronaviruses (4.2%). In 2015, additional pathogens were investigated with the following detection rate: enterovirus (13.2%), bocavirus (HBoV) (10.5%), PIV-4 (2.6%), and parechovirus (1.3%). There were no statistical differences between single and multiple virus infection regarding patients age, localization of infection, and severity of disease . AdV, HRV, HBoV, and PIVs were significantly more often detected in multiple virus infections compared to the other respiratory viruses . Sunčanica Ljubin-Sternak, Tatjana Marijan, Irena Ivković-Jureković, Jasna Čepin-Bogović, Alenka Gagro, and Jasmina Vraneš Copyright © 2016 Sunčanica Ljubin-Sternak et al. All rights reserved. Clinical Effects of Gamma-Radiation-Resistant Aspergillus sydowii on Germ-Free Mice Immunologically Prone to Inflammatory Bowel Disease Thu, 18 Aug 2016 16:01:27 +0000 We report and investigated a case of inadvertent contamination of 125 mice (housed in two germ-free positive-pressurized isolators) with emerging human and coral pathogen Aspergillus sydowii. The infected mice correspond to genetic line SAMP1/YitFc, which have 100% immune predisposition to develop Crohn’s disease-like spontaneous pathologies, namely, inflammatory bowel disease (IBD). Pathogen update based on a scoping review of the literature and our clinical observations and experimentation are discussed. The unwanted infection of germ-free mice (immunologically prone to suffer chronic inflammation) with human pathogen A. sydowii resulted in no overt signs of clinical disease over 3-week exposure period, or during DSS-induced colitis experiments. Results and observations suggest that A. sydowii alone has limited clinical effect in immunocompromised germ-free mice or that other commensal microbial flora is required for Aspergillus-associated disease to occur. Alexander Rodriguez-Palacios, Natalia Aladyshkina, Mauricio Retuerto, Christopher L. Hager, Sanja Ilic, Mahmoud A. Ghannoum, and Fabio Cominelli Copyright © 2016 Alexander Rodriguez-Palacios et al. All rights reserved. Prevalence of Tinea Capitis among School Children in Nok Community of Kaduna State, Nigeria Mon, 04 Jul 2016 12:31:03 +0000 In recent years, the prevalence of tinea capitis, an infection of the scalp by dermatophytes, has increased in children worldwide. This cross-sectional study was carried out to determine the prevalence and risk factor of tinea capitis among school children in Nok community of Kaduna State, Nigeria. A total of 100 children were screened and 45% were diagnosed to have tinea capitis after fungal culture and microscopy. The prevalence of tinea capitis among girls was higher (51.4%) than that among boys (41.5%) but not significantly different (). The prevalence with respect to age was lower for the age group 5–10 years (42.6%) than that of 11–15 years (50%) but was not significantly different (). Trichophyton rubrum (28.8%) and Microsporum canis (22.7%) were the most prevalent dermatophytes isolated and the least were Trichophyton verrucosum (4.5%) and Trichophyton tonsurans (4.5%). There were 73.3% single infection while 26.7% had 2–4 dermatophytes of the genera Microsporum and Trichophyton. The predisposing factors with statistically significant association with tinea capitis were number of children in the family () and sharing of the same bed (). This indicates the high tendencies of spread of tinea capitis through human-to-human mode of transmission and possible animal contact. Community health education on the cause, mode of transmission, prevention, and prompt treatment of tinea capitis is recommended. Josephine Dogo, Seniyat Larai Afegbua, and Edward Christopher Dung Copyright © 2016 Josephine Dogo et al. All rights reserved. PCR-Based Molecular Diagnosis of Hepatitis Virus (HBV and HDV) in HCV Infected Patients and Their Biochemical Study Mon, 06 Jun 2016 11:50:22 +0000 Seroprevalence of HCV indicates that HCV is found in more than 10% of HBV- or HDV-infected patients worldwide leading to liver disease. Here we show HBV and HDV coinfection association with HCV infected Pakistani patients, study of disease severity, and possible interpretation of associated risk factors in coinfected patients. A total of 730 liver diseased patients were included, out of which 501 were found positive for HCV infection via PCR. 5.1% of patients were coinfected with HBV while 1% were coinfected with HBV and HDV both. LFTs were significantly altered in dually and triply infected patients as compared to single HCV infection. Mean bilirubin, AST, and ALT levels were highest (3.25 mg/dL, 174 IU/L, and 348 IU/L) in patients with triple infection while dual infection LFTs (1.6 mg/dL, 61 IU/L, and 74 IU/L) were not high as in single infection (1.9 mg/dL, 76 IU/L, and 91 IU/L). The most prominent risk factor in case of single (22%) and dual infection (27%) group was “reuse of syringes” while in triple infection it was “intravenous drug users” (60%). It is concluded that HBV and HDV coinfections are strongly associated with HCV infected Pakistani patients and in case of severe liver disease the possibility of double and triple coinfection should be kept in consideration. Muhammad Nasir Riaz, Muhammad Faheem, Muhammad Ayaz Anwar, Ummar Raheel, Yasmeen Badshah, Hashaam Akhtar, Kosar Tamanna, Muhammad Tahir, Najam us Sahar Sadaf Zaidi, and Ishtiaq Qadri Copyright © 2016 Muhammad Nasir Riaz et al. All rights reserved. A Survey of Seasonal Gastrointestinal Parasitic Infections in Donkeys from a Semiarid Sub-Saharan Region, Sudan Thu, 19 May 2016 08:38:02 +0000 Out of 92 donkeys examined for gastrointestinal parasites, 90 animals were found infected by one or more gastrointestinal parasites with an overall prevalence rate of 97.78%. The distributions of the recovered parasites in the different parts of the body were as follows: stomach, 92.4%, small intestine, 19.6%, caecum, 88%, colon, 80.4%, rectum, 73.9%, and cranial mesenteric artery, 64.1%. A significant difference was found between mean parasite counts and seasons. Hot wet season had higher mean parasites count () in comparison with hot dry () and cool dry () seasons. Although there was no significant difference between age and mean parasite count, animals more than four years old had high mean count () in comparison with and for young and adults animals, respectively. No significant positive or negative correlation was found between total parasite counts of infected animals and any of the climatic factors. The parasites identified were Habronema spp. (40.2%), Trichostrongylus axei (30.4%), Parascaris equorum (18.5%), Anoplocephala perfoliata (4.35%), Gastrodiscus aegyptiacus (8.7%), large strongyles (84%), small strongyles (72%), and Oxyuris equi (1.1%). Ahmed Abdurhman Ismail, Nasredin Khogali Ahmed, Ahmed Elhag Bashar, Hisham Ismail Seri, El Tigani Ahmed El Tigani-Asil, and Adam Dawoud Abakar Copyright © 2016 Ahmed Abdurhman Ismail et al. All rights reserved. Antimicrobial Activity of Croton macrostachyus Stem Bark Extracts against Several Human Pathogenic Bacteria Wed, 11 May 2016 13:28:15 +0000 In Kenya, leaves and roots from Croton macrostachyus are used as a traditional medicine for infectious diseases such as typhoid and measles, but reports on possible antimicrobial activity of stem bark do not exist. In this study, the antibacterial and antifungal effects of methanol, ethyl acetate and butanol extracts, and purified lupeol of C. macrostachyus stem bark were determined against important human gram-negative pathogens Escherichia coli, Salmonella typhi, Klebsiella pneumoniae, and Enterobacter aerogenes, gram-positive Listeria monocytogenes, and a fungus Candida albicans. The most promising broad scale antimicrobial activity against all the studied pathogens was shown by the ethyl acetate extract. The ethyl acetate extract induced the zone of inhibition between  mm and  mm against S. typhi, E. coli, K. pneumoniae, E. aerogenes, and L. monocytogenes with weaker antimicrobial activity against C. albicans (zone of inhibition:  mm). The antibiotic controls (amoxicillin, ciprofloxacin, ampicillin, benzylpenicillin, clotrimazole, and cefotaxime) showed antimicrobial activity with zones of inhibition within  mm. The ethyl acetate extract had MIC in the range of 125–250 mg/mL against all the studied bacteria and against C. albicans MIC was 500 mg/mL. The present results give scientific evidence and support the traditional use of C. macrostachyus stem bark as a source for antimicrobials. We show that C. macrostachyus stem bark lupeol is a promising antimicrobial agent against several important human pathogens. Jackie K. Obey, Atte von Wright, Jimmy Orjala, Jussi Kauhanen, and Carina Tikkanen-Kaukanen Copyright © 2016 Jackie K. Obey et al. All rights reserved. Virulence Genes among Enterococcus faecalis and Enterococcus faecium Isolated from Coastal Beaches and Human and Nonhuman Sources in Southern California and Puerto Rico Sun, 10 Apr 2016 07:55:37 +0000 Most Enterococcus faecalis and E. faecium are harmless to humans; however, strains harboring virulence genes, including esp, gelE, cylA, asa1, and hyl, have been associated with human infections. E. faecalis and E. faecium are present in beach waters worldwide, yet little is known about their virulence potential. Here, multiplex PCR was used to compare the distribution of virulence genes among E. faecalis and E. faecium isolated from beaches in Southern California and Puerto Rico to isolates from potential sources including humans, animals, birds, and plants. All five virulence genes were found in E. faecalis and E. faecium from beach water, mostly among E. faecalis. gelE was the most common among isolates from all source types. There was a lower incidence of asa1, esp, cylA, and hyl genes among isolates from beach water, sewage, septage, urban runoff, sea wrack, and eelgrass as compared to human isolates, indicating that virulent strains of E. faecalis and E. faecium may not be widely disseminated at beaches. A higher frequency of asa1 and esp among E. faecalis from dogs and of asa1 among birds (mostly seagull) suggests that further studies on the distribution and virulence potential of strains carrying these genes may be warranted. Donna M. Ferguson, Ginamary Negrón Talavera, Luis A. Ríos Hernández, Stephen B. Weisberg, Richard F. Ambrose, and Jennifer A. Jay Copyright © 2016 Donna M. Ferguson et al. All rights reserved. The Efficacy of Umbelliferone, Arbutin, and N-Acetylcysteine to Prevent Microbial Colonization and Biofilm Development on Urinary Catheter Surface: Results from a Preliminary Study Tue, 05 Apr 2016 11:36:55 +0000 We evaluated, in a preliminary study, the efficacy of umbelliferone, arbutin, and N-acetylcysteine to inhibit biofilm formation on urinary catheter. We used 20 urinary catheters: 5 catheters were incubated with Enterococcus faecalis (control group); 5 catheters were incubated with E. faecalis in presence of umbelliferone (150 mg), arbutin (60 mg), and N-acetylcysteine (150 mg) (group 1); 5 catheters were incubated with E. faecalis in presence of umbelliferone (150 mg), arbutin (60 mg), and N-acetylcysteine (400 mg) (group 2); and 5 catheters were incubated with E. faecalis in presence of umbelliferone (300 mg), arbutin (60 mg), and N-acetylcysteine (150 mg) (group 3). After 72 hours, planktonic microbial growth and microorganisms on catheter surface were assessed. In the control group, we found a planktonic load of ≥105 CFU/mL in the inoculation medium and retrieved 3.69 × 106 CFU/cm from the sessile cells adherent to the catheter surface. A significantly lower amount in planktonic () and sessile () bacterial load was found in group 3, showing <100 CFU/mL and 0.12 × 106 CFU/cm in the incubation medium and on the catheter surface, respectively. In groups 1 and 2, 1.67 × 106 CFU/cm and 1.77 × 106 CFU/cm were found on catheter surface. Our results document that umbelliferone, arbutin, and N-acetylcysteine are able to reduce E. faecalis biofilm development on the surface of urinary catheters. Tommaso Cai, Luca Gallelli, Francesca Meacci, Anna Brugnolli, Letizia Prosperi, Stefani Roberta, Cristina Eccher, Sandra Mazzoli, Paolo Lanzafame, Patrizio Caciagli, Gianni Malossini, and Riccardo Bartoletti Copyright © 2016 Tommaso Cai et al. All rights reserved. Phenotypic Detection of Metallo-Beta-Lactamases in Carbapenem Resistant Acinetobacter baumannii Isolated from Pediatric Patients in Pakistan Wed, 30 Mar 2016 14:22:59 +0000 Multidrug resistant A. baumannii has emerged as an important and problematic human pathogen as it is the causative agent of several types of infections especially in neonates and immunocompromised patients because they have least capacity to fight against infections. Carbapenems are used as last resort antibiotics for treating these infections but currently resistance against carbapenems due to MBL production is on the rise. The objective of this study was to determine the frequency of antibiotic resistance in A. baumannii and also to compare the efficacy of combined disk test and double disk synergy test for detection of metallo-beta-lactamases. A total of 112 A. baumannii were identified from various clinical samples and antibiotic susceptibility profile was determined by Kirby-Bauer Disk Diffusion method. Out of 112, 66 (58.9%) isolates were resistant to both imipenem and meropenem (OXOID). These resistant isolates were tested for carbapenemase production, and 55 (83.3%) were carbapenemase producers by Modified Hodge Test. These isolates were further tested for MBL production by combined disk test and double disk synergy test. Out of 66, 49 isolates were positive by both methods, CDT and DDST, and only one isolate was detected as negative (with kappa value = 0.038). All MBL producing strains showed remarkable resistance to cephalosporins, fluoroquinolones, aminoglycosides, and piperacillin/tazobactam (OXOID). The antibiotic resistance was very high in A. baumannii which were isolated from children in Pakistan specially attending a nephrology unit. Muneeza Anwar, Hassan Ejaz, Aizza Zafar, and Hamdan Hamid Copyright © 2016 Muneeza Anwar et al. All rights reserved. Multidrug Resistant CTX-M-Producing Escherichia coli: A Growing Threat among HIV Patients in India Wed, 30 Mar 2016 07:00:02 +0000 Extended Spectrum β-Lactamases (ESBLs) confer resistance to third-generation cephalosporins and CTX-M types have emerged as the most prominent ESBLs worldwide. This study was designed to determine the prevalence of CTX-M positive ESBL-producing urinary E. coli isolates from HIV patients and to establish the association of multidrug resistance, phylogeny, and virulence profile with CTX-M production. A total of 57 ESBL producers identified among 76 E. coli strains isolated from HIV patients from South India were screened for , AmpC production, multidrug resistance, and nine virulence associated genes (VAGs), fimH, pap, afa/dra, sfa/foc, iutA, fyuA, iroN, usp, and kpsMII. The majority (70.2%) of the ESBL producers harbored and were AmpC coproducers. Among the CTX-M producers, 47.5% were found to be UPEC, 10% harbored as many as 7 VAGs, and 45% possessed kpsMII. Multidrug resistance (CIPRSXTRGENR) was significantly more common among the CTX-M producers compared to the nonproducers (70% versus 41.2%). However, 71.4% of the multidrug resistant CTX-M producers exhibited susceptibility to nitrofurantoin thereby making it an effective alternative to cephalosporins/fluoroquinolones. The emergence of CTX-M-producing highly virulent, multidrug resistant uropathogenic E. coli is of significant public health concern in countries like India with a high burden of HIV/AIDS. Kesavaram Padmavathy, Krishnan Padma, and Sikhamani Rajasekaran Copyright © 2016 Kesavaram Padmavathy et al. All rights reserved. Nontuberculous Mycobacteria Isolated from Tuberculosis Suspects in Ibadan, Nigeria Wed, 23 Mar 2016 12:38:01 +0000 In Nigeria, one of the highest tuberculosis (TB) burdened nations, sputum smear microscopy is routinely employed for TB diagnosis at Directly Observed Treatment Short-Course (DOTS) Centers. This diagnostic algorithm does not differentiate Mycobacterium tuberculosis complex (MTC) from nontuberculous mycobacteria (NTM). Between December 2008 and January 2009, consecutive patients diagnosed with TB were screened for inclusion at 10 DOTS centers in Ibadan, Nigeria. To verify Mycobacterium species in patients diagnosed, we cultured and identified mycobacterial isolates using PCR, line probe assay, and spoligotyping techniques. From 48 patients screened, 23 met the inclusion criteria for the study. All the 23 study patients had a positive culture. Overall, we identified 11/23 patients (48%) with MTC only, 9/23 (39%) with NTM only, and 3/23 (13%) with evidence of both MTC and NTM. Strains of MTC identified were Latin American Mediterranean (LAM) genotype (), M. africanum (), and the genotype family T (). Four M. avium-intracellulare-M. scrofulaceum complexes, one M. chelonae complex, one M. abscessus, and one M. intracellulare were identified. Our findings underscore the need to incorporate molecular techniques for more precise diagnosis of TB at DOTS centers to improve clinical outcomes and safe guard public health, particularly in TB endemic countries. Simeon Idowu Cadmus, Bassirou Diarra, Brehima Traore, Mamoudou Maiga, Sophia Siddiqui, Anatole Tounkara, Olutayo Falodun, Wole Lawal, Isaac Folurunso Adewole, Rob Murphy, Dick van Soolingen, and Babafemi Taiwo Copyright © 2016 Simeon Idowu Cadmus et al. All rights reserved. Primary Multidrug Resistant Tuberculosis and Utility of Line Probe Assay for Its Detection in Smear-Positive Sputum Samples in a Tertiary Care Hospital in South India Wed, 23 Mar 2016 12:19:48 +0000 In a high tuberculosis burdened country like India, rapid, cost-effective, and reliable diagnostic tools for tuberculosis are an urgent need of the hour to prevent inappropriate treatment strategies and further spread of resistance. This study aimed to estimate the proportion of new smear-positive tuberculosis cases with primary resistance to rifampicin and/or isoniazid as well as identify the common mutations associated with it. Sputum of 200 newly diagnosed smear-positive cases of 1+ score and above was directly subjected to Line Probe Assay using the GenoType MTBDRplus assay kit. All samples were inoculated onto solid media and 61 samples were inoculated in automated liquid culture also. The Line Probe Assay gave hundred percent interpretable results with 2.5% of the study population showing resistant pattern. Only 1% of the cases were primary multidrug resistant tuberculosis and 1.5% showed isoniazid monoresistance. S531L and C15T were the most common genetic mutations seen for rifampicin and isoniazid resistance, respectively. 40% had absent rpoB wild type 8 band indicating probable silent mutation after clinical correlation. The average turnaround time for Line Probe Assay was far less (3.8 days) as compared to solid and liquid cultures (35.6 days and 13.5 days, resp.). Fahmiya Leena Yacoob, Beena Philomina Jose, Sarada Devi Karunakaran Lelitha, and Sreelatha Sreenivasan Copyright © 2016 Fahmiya Leena Yacoob et al. All rights reserved. In Vivo Antimalarial Activity of Annona muricata Leaf Extract in Mice Infected with Plasmodium berghei Tue, 22 Mar 2016 14:23:03 +0000 Malaria is one of the most important infectious diseases in the world. The choice for the treatment is highly limited due to drug resistance. Hence, finding the new compounds to treat malaria is urgently needed. The present study was attempted to evaluate the antimalarial activity of the Annona muricata aqueous leaf extract in Plasmodium berghei infected mice. Aqueous leaf extract of A. muricata was prepared and tested for acute toxicity in mice. For efficacy test in vivo, standard 4-day suppressive test was carried out. ICR mice were inoculated with 107 parasitized erythrocytes of P. berghei ANKA by intraperitoneal injection. The extracts (100, 500, and 1000 mg/kg) were then given orally by gavage once a day for 4 consecutive days. Parasitemia, percentage of inhibition, and packed cell volume were subsequently calculated. Chloroquine (10 mg/kg) was given to infected mice as positive control while untreated control was given only distilled water. It was found that A. muricata aqueous leaf extract at doses of 100, 500, and 1000 mg/kg resulted in dose dependent parasitemia inhibition of 38.03%, 75.25%, and 85.61%, respectively. Survival time was prolonged in infected mice treated with the extract. Moreover, no mortality to mice was observed with this extract up to a dose of 4000 mg/kg. In conclusion, the A. muricata aqueous leaf extract exerted significant antimalarial activity with no toxicity and prolonged survival time. Therefore, this extract might contain potential lead molecule for the development of a new drug for malaria treatment. Voravuth Somsak, Natsuda Polwiang, and Sukanya Chachiyo Copyright © 2016 Voravuth Somsak et al. All rights reserved. Candida Species Prevalence Profile in HIV Seropositive Patients from a Major Tertiary Care Hospital in New Delhi, India Tue, 22 Mar 2016 08:50:26 +0000 Candida is a common opportunistic pathogen during the course of human immunodeficiency virus (HIV) disease progression. Changes in the clinical severity of candidiasis and the Candida species prevalence profile may be a reflection of immunological changes in HIV positive patients. The aim of this study was to document the changing pattern of Candida species prevalence profile in HIV seropositive patients from a tertiary care hospital in North India. One hundred and twenty HIV seropositive subjects were recruited for Candida microbial screening. Clinical specimens including blood, oral swabs, expectorated or induced sputum/bronchoalveolar lavage specimens, and urine were collected depending on the patient’s symptoms. A total of 128 Candida isolates were obtained from 88 cases and 7 different Candida species were identified. C. albicans (50%) was the most common species isolated followed by C. glabrata (17%) and C. dubliniensis (12.5%). Other species isolated were C. parapsilosis (7.8%), C. krusei, C. tropicalis (4.6% each), and C. kefyr (3%). Strong clinical suspicion along with optimal sampling of an accurate diagnosis of Candida species involved would go a long way in decreasing the morbidity associated with non-albicans Candida species. Monika Maheshwari, Ravinder Kaur, and Sanjim Chadha Copyright © 2016 Monika Maheshwari et al. All rights reserved. Serotyping and Antimicrobial Susceptibility Pattern of Escherichia coli Isolates from Urinary Tract Infections in Pediatric Population in a Tertiary Care Hospital Mon, 07 Mar 2016 11:09:46 +0000 Urinary tract infections (UTIs) in pediatric population are associated with high morbidity and long term complications. In recent years, there is increased prevalence of Escherichia coli (E. coli) strains producing extended spectrum β-lactamase, Amp C, and Metallo β-lactamase, making the clinical management even more difficult. This study was aimed to detect the serotypes and to determine antimicrobial susceptibility profile of E. coli isolates from urine samples of children <10 yrs old. A total of 75 pure E. coli strains isolated from patients with symptoms of UTI and colony count ≥105 organisms/mL were included in the study. Antibiotic sensitivity pattern showed maximum resistance to nalidixic acid (98.7%), followed by ampicillin (97.3%), amoxi-clavulanate (96%), and fluoroquinolones (92%) while most of the isolates were found sensitive to piperacillin-tazobactam (13.3%), nitrofurantoin (5.3%), and meropenem (1.3%). 48% of the strains were ESBL producer (extended spectrum beta lactamase). 44% strains were typable withantisera used in our study and the most common serogroup was O6 (33.3%) followed by O1 (15.1%) and O15 (15.1%). To conclude, judicious use of antibiotics according to hospital antibiotic policy and infection control measures should be implemented to prevent spread of multidrug resistant organisms. Shweta Sharma, Nirmaljit Kaur, Shalini Malhotra, Preeti Madan, Wasim Ahmad, and Charoo Hans Copyright © 2016 Shweta Sharma et al. All rights reserved. Vancomycin and High Level Aminoglycoside Resistance in Enterococcus spp. in a Tertiary Health Care Centre: A Therapeutic Concern Mon, 07 Mar 2016 09:57:41 +0000 Aims. This study was aimed at knowing the prevalence of vancomycin and high level aminoglycoside resistance in enterococcal strains among clinical samples. Study Design. It was an investigational study. Place and Duration of Study. It was conducted on 100 Enterococcus isolates, in the Department of Microbiology, Pt. BDS PGIMS, Rohtak, over a period of six months from July to December 2014. Methodology. Clinical specimens including urine, pus, blood, semen, vaginal swab, and throat swab were processed and Enterococcus isolates were identified by standard protocols. Antibiotic sensitivity testing of enterococci was performed using Kirby-Bauer disc diffusion method. Results. High level gentamicin resistance (HLGR) was more common in urine samples (41.5%) followed by blood (36%) samples. High level streptomycin resistance (HLSR) was more common in pus samples (52.6%) followed by blood samples (36%). Resistance to vancomycin was maximum in blood isolates. Conclusion. Enterococci resistant to multiple antimicrobial agents have been recognized. Thus, it is crucial for laboratories to provide accurate antimicrobial resistance patterns for enterococci so that effective therapy and infection control measures can be initiated. Seema Mittal, Pooja Singla, Antariksha Deep, Kiran Bala, Rama Sikka, Meenu Garg, and Uma Chaudhary Copyright © 2016 Seema Mittal et al. All rights reserved.