Journal of Pathogens The latest articles from Hindawi Publishing Corporation © 2016 , Hindawi Publishing Corporation . 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. Antibiotic Prophylaxis for Gynecologic Procedures prior to and during the Utilization of Assisted Reproductive Technologies: A Systematic Review Mon, 07 Mar 2016 08:06:43 +0000 The use of assisted reproductive technologies (ART) has increased steadily. There has been a corresponding increase in the number of ART-related procedures such as hysterosalpingography (HSG), saline infusion sonography (SIS), hysteroscopy, laparoscopy, oocyte retrieval, and embryo transfer (ET). While performing these procedures, the abdomen, upper vagina, and endocervix are breached, leading to the possibility of seeding pelvic structures with microorganisms. Antibiotic prophylaxis is therefore important to prevent or treat any procedure-related infections. After careful review of the published literature, it is evident that routine antibiotic prophylaxis is generally not recommended for the majority of ART-related procedures. For transcervical procedures such as HSG, SIS, hysteroscopy, ET, and chromotubation, patients at risk for pelvic infections should be screened and treated prior to the procedure. Patients with a history of pelvic inflammatory disease (PID) or dilated fallopian tubes are at high risk for postprocedural infections and should be given antibiotic prophylaxis during procedures such as HSG, SIS, or chromotubation. Antibiotic prophylaxis is recommended prior to oocyte retrieval in patients with a history of endometriosis, PID, ruptured appendicitis, or multiple prior pelvic surgeries. Nigel Pereira, Anne P. Hutchinson, Jovana P. Lekovich, Elie Hobeika, and Rony T. Elias Copyright © 2016 Nigel Pereira et al. All rights reserved. Genotype Cluster Analysis in Pathogenic Escherichia coli Isolates Producing Different CDT Types Thu, 03 Mar 2016 07:49:20 +0000 Diarrheagenic and uropathogenic E. coli types are mainly characterized by the expression of distinctive bacterial virulent factors. stx1, stx2 (Shiga toxins), and cdt (cytolethal distending toxin) genes have been acquired by horizontal gene transfer. Some virulent genes such as espP (serine protease), etpD (part of secretion pathway), and katP (catalase-peroxidase), or sfpA gene (Sfp fimbriae), are on plasmids and the others like fliC (flagellin) and the fimH gene (fimbriae type-I) are located on chromosome. Genomic pathogenicity islands (PAIs) carry some virulent genes such as hly gene. To determine the existence of virulence genes in cdt clinical isolates, genes including stx1, stx2, cdt, hly, espP, katP, sfpA, etpD, fliC, and fimH were assessed by Polymerase Chain Reaction (PCR). The most prevalent isolates for etpD and katP genes were 85.7% in cdtII. katP gene was also observed 83.3% in cdtI. However, in 42.85% of cdtIII isolates, espP gene was the most detected. Moreover, hly gene was also the most prominent gene in cdtIII (71.42%). sfpA gene was observed in 66.6% of cdtV. stx1 gene was detected in 100% of cdtII, cdtIV, and cdtV types. Presence and pattern of virulence genes were considered among cdt positive isotypes and used for their clustering and profiling. Maryam Javadi, Mana Oloomi, and Saeid Bouzari Copyright © 2016 Maryam Javadi et al. All rights reserved. Encephalitozoon cuniculi: Grading the Histological Lesions in Brain, Kidney, and Liver during Primoinfection Outbreak in Rabbits Sun, 28 Feb 2016 12:46:58 +0000 This is the first confirmed report of Encephalitozoon cuniculi (E. cuniculi) in farm meat rabbits located in Northern Mexico. Eighty young rabbits exhibited clinical signs of this zoonotic emerging disease, like torticollis, ataxia, paresis, circling, and rolling. Samples of brain, kidney, and liver were examined for histology lesions. For the first time the lesions caused by E. cuniculi were graded according to their severity (I, II, and III) and the size of the granulomas (Types A, B, and C). The main cerebral injuries were Grade III, coinciding with the presence of Type C granulomas. The cerebral lesions were located in the cortex, brain stem, and medulla. The renal lesions were also Grade III distributed throughout cortex and renal medulla, with no granuloma formation. The involvement of hypersensitivity Types III and IV is suggested. All of the rabbits were seropositive to E. cuniculi by CIA testing, suggesting that this zoonotic and emerging pathogen is widely distributed among animals intended for human consumption. We believe this work could be used as a guide when examining E. cuniculi and will provide direction to confirm the diagnosis of this pathogen. Luis E. Rodríguez-Tovar, Alicia M. Nevárez-Garza, Armando Trejo-Chávez, Carlos A. Hernández-Martínez, Gustavo Hernández-Vidal, Juan J. Zarate-Ramos, and Uziel Castillo-Velázquez Copyright © 2016 Luis E. Rodríguez-Tovar et al. All rights reserved. Reproductive Pathological Changes Associated with Experimental Subchronic Corynebacterium pseudotuberculosis Infection in Nonpregnant Boer Does Tue, 23 Feb 2016 16:15:24 +0000 Corynebacterium pseudotuberculosis causes caseous lymphadenitis (CLA), which is a contagious and chronic disease in sheep and goats. In order to assess the histopathological changes observed in the reproductive organs of nonpregnant does infected with the bacteria, 20 apparently healthy adult Boer does were divided into four inoculation groups, intradermal, intranasal, oral, and control, consisting of five goats each. Excluding the control group, which was unexposed, other does were inoculated with 107 CFU/1 mL of live C. pseudotuberculosis through the various routes stated above. Thirty days after infection, the ovaries, uterus, and iliac lymph nodes were collected for bacterial recovery and molecular detection, as well as histopathological examination. The mean changes in necrosis, congestion, inflammatory cell infiltration, and oedema varied in severity among the ovaries, uterus, and iliac lymph nodes following different inoculation routes. Overall, the intranasal route of inoculation showed more severe () lesions in all the organs examined. The findings of this study have shown that C. pseudotuberculosis could predispose to infertility resulting from pathological lesions in the uterus and ovaries of does. A. M. Othman, Y. Abba, F. F. A. Jesse, Y. M. Ilyasu, A. A. Saharee, A. W. Haron, M. Zamri-Saad, and M. A. M. Lila Copyright © 2016 A. M. Othman et al. All rights reserved. Genotyping of Cryptosporidium Species and Their Clinical Manifestations in Patients with Renal Transplantation and Human Immunodeficiency Virus Infection Sun, 14 Feb 2016 12:03:09 +0000 In the present study we aimed to determine (i) frequency of Cryptosporidium species among patients with renal transplantation (RT) and human immunodeficiency virus (HIV) infection and (ii) relationship of the nature, severity, and duration of symptoms with different species and load of Cryptosporidium. Stool samples from 70 (42 RT and 28 HIV) and 140 immunocompromised patients with and without cryptosporidiosis by modified Kinyoun’s staining were subjected to qPCR-melting curve analysis for identification of parasite species. qPCR detected one microscopically negative sample to be positive for cryptosporidiosis. C. hominis, C. parvum, and mixed infection were detected in 50/71 (70.4%), 19/71 (26.8%), and 2/71 (2.8%) patients, respectively. Patients with cryptosporidiosis had higher stool frequency (median, IQR: 4, 3–6/d versus 3, 2–4/d; ) and watery stool (52/71 [73%] versus 64/139 [46%]; ). Parasite load (median, IQR: Log10 6.37 (5.65–7.12), Log10 5.81 (4.26–6.65); ) and nausea/vomiting (29/50 [58%] versus 5/19 [26%]; ) were more frequent with C. hominis than with C. parvum infection. Thus, Cryptosporidium spp. (mainly C. hominis) is a common cause of diarrhoea in RT and HIV patients. Asmita Dey, Ujjala Ghoshal, Vikas Agarwal, and Uday Chand Ghoshal Copyright © 2016 Asmita Dey et al. All rights reserved. Antibiotic-Resistant Acinetobacter baumannii Increasing Success Remains a Challenge as a Nosocomial Pathogen Thu, 04 Feb 2016 07:09:42 +0000 Antibiotic-resistant infectious bacteria currently imply a high risk and therefore constitute a strong challenge when treating patients in hospital settings. Characterization of these species and of particular strains is a priority for the establishment of diagnostic tests and preventive procedures. The relevance of Acinetobacter baumannii as a problematic microorganism in inpatient facilities, particularly intensive care units, has increased over time. This review aims to draw attention to (i) the historical emergence of carbapenem-resistant Acinetobacter baumannii, (ii) the current status of surveillance needs in Latin America, and (iii) recent data suggesting that A. baumannii continues to spread and evolve in hospital settings. First, we present synopsis of the series of events leading to the discovery and precise identification of this microorganism in hospital settings. Then key events in the acquisition of antibiotic-resistant genes by this microorganism are summarized, highlighting the race between new antibiotic generation and emergence of A. baumannii resistant strains. Here we review the historical development of this species as an infectious threat, the current state of its distribution, and antibiotic resistance characteristics, and we discuss future prospects for its control. Ana Maria Gonzalez-Villoria and Veronica Valverde-Garduno Copyright © 2016 Ana Maria Gonzalez-Villoria and Veronica Valverde-Garduno. All rights reserved. Multidrug Resistant and Extensively Drug Resistant Bacteria: A Study Thu, 28 Jan 2016 08:50:45 +0000 Background and Objective. Antimicrobial resistance is now a major challenge to clinicians for treating patients. Hence, this short term study was undertaken to detect the incidence of multidrug-resistant (MDR), extensively drug-resistant (XDR), and pandrug-resistant (PDR) bacterial isolates in a tertiary care hospital. Material and Methods. The clinical samples were cultured and bacterial strains were identified in the department of microbiology. The antibiotic susceptibility profile of different bacterial isolates was studied to detect MDR, XDR, and PDR bacteria. Results. The antibiotic susceptibility profile of 1060 bacterial strains was studied. 393 (37.1%) bacterial strains were MDR, 146 (13.8%) strains were XDR, and no PDR was isolated. All (100%) Gram negative bacterial strains were sensitive to colistin whereas all (100%) Gram positive bacterial strains were sensitive to vancomycin. Conclusion. Close monitoring of MDR, XDR, or even PDR must be done by all clinical microbiology laboratories to implement effective measures to reduce the menace of antimicrobial resistance. Silpi Basak, Priyanka Singh, and Monali Rajurkar Copyright © 2016 Silpi Basak et al. All rights reserved. Prevalence of Methicillin Resistant Staphylococcal Bioaerosols in and around Residential Houses in an Urban Area in Central India Tue, 26 Jan 2016 06:38:46 +0000 Methicillin resistant staphylococci (MRS) commonly found in clinical samples or associated environment pose a major health challenge globally. The carriage rate of MRS in human population is high, especially in India but research on airborne distribution of MRS is scanty. The present study aimed to evaluate the prevalence of MRS in indoor and outdoor environment of residential houses. Air samples were collected using impactor air sampler. The total counts of viable bacteria, staphylococci, and MRS along with the particles of various sizes were determined from indoor and outdoor environment of 14 residential houses. MRS bacteria were identified as methicillin resistant S. aureus (MRSA) or coagulase negative staphylococci (CNS) employing biochemical and PCR assays. The average concentration of MRS inside and outside of the houses was 5.9% and 4.6% of the total bacteria, respectively. The maximum correlation of total indoor and outdoor bacteria with particulate matter was 10 μm () and 5 μm (), respectively. Statistically, significant positive correlation of staphylococci and MRS was found with particles of 10–25 μm inside the houses. Molecular surveillance, antibiotic stewardship programme, and infection control policies can help to manage increasing MRS burden in developing countries. P. Kumar and A. K. Goel Copyright © 2016 P. Kumar and A. K. Goel. All rights reserved. Emergence of Tetracycline Resistant Vibrio cholerae O1 Biotype El Tor Serotype Ogawa with Classical ctxB Gene from a Cholera Outbreak in Odisha, Eastern India Sun, 03 Jan 2016 14:06:22 +0000 In September 2010, a cholera outbreak was reported from Odisha, Eastern India. V. cholerae isolated from the clinical samples were biochemically and serologically confirmed as serogroup O1, biotype El Tor, and serotype Ogawa. Multiplex PCR screening revealed the presence of various genes, namely, ompW, ctxB, zot, rfbO1, tcp, ace, hlyA, ompU, rtx, and toxR, in all of the isolates. The isolates were resistant to co-trimoxazole, nalidixic acid, polymyxin B, spectinomycin, streptomycin, sulfamethoxazole, tetracycline, trimethoprim, and vibriostatic agent 2,4-diamino-6,7-diisopropylpteridine (O/129). Minimum inhibitory concentration of tetracycline decreased in the presence of carbonyl cyanide m-chlorophenylhydrazone (CCCP), suggesting the involvement of efflux pumps. PCR analysis confirmed the presence of class I integrons as well as SXT elements harbouring antibiotic resistance genes in all isolates. Sequencing revealed the presence of ctxB gene of classical biotype in all the isolates. The isolates harboured an RS1-CTX prophage array with El Tor type rstR and classical ctxB on the large chromosome. The study indicated that the V. cholerae El Tor variants are evolving in the area with better antibiotic resistance and virulence potential. M. Jain, P. Kumar, and A. K. Goel Copyright © 2016 M. Jain et al. All rights reserved. Susceptibility Pattern and Distribution of Oxacillinases and blaPER-1 Genes among Multidrug Resistant Acinetobacter baumannii in a Teaching Hospital in Iran Thu, 31 Dec 2015 08:19:51 +0000 Acinetobacter baumannii (A. baumannii) is an important nosocomial pathogen in healthcare institutions. β-Lactamase-mediated resistance is the most common mechanism for carbapenem resistance in A. baumannii. The aim of this study was to determine the antibiotic resistance pattern, to detect OXA encoding genes, class A, , and to detect the presence of ISAba1. A total of 124 A. baumannii isolates were collected from hospitalized patients in a teaching hospital in Kashan, Iran. The susceptibility of isolates to different antibiotics was determined by disk-diffusion method. PCR was used to detect , , , , , and ISAba1 genes. All isolates were resistant to ceftazidime, ceftriaxone, and cefotaxime. All of the isolates revealed susceptibility to polymyxin B and colistin. Ninety-six percent of the isolates were extensive drug resistance (XDR), 5.6% extended spectrum beta-lactamase (ESBL), and 54.8% metallo-beta-lactamase (MBL). All isolates were positive for and ISAba1. ,, and were found in 79.8%, 25%, and 3.2%, respectively. The frequency rate of gene was 52.4%. Multidrug resistant A. baumannii isolates are increasing in our setting and extensively limit therapeutic options. The high rate presence of class D carbapenemase-encoding genes, mainly carbapenemases, is worrying and alarming as an emerging threat in our hospital. Sareh Bagheri Josheghani, Rezvan Moniri, Farzaneh Firoozeh, Mojtaba Sehat, and Yasaman Dasteh Goli Copyright © 2015 Sareh Bagheri Josheghani et al. All rights reserved. Isolation and Antibiotic Susceptibility of the Microorganisms Isolated from Diabetic Foot Infections in Nemazee Hospital, Southern Iran Wed, 30 Dec 2015 12:48:42 +0000 Background. Diabetic foot infections (DFIs) are a major public health issue and identification of the microorganisms causing such polymicrobial infections is useful to find out appropriate antibiotic therapy. Meanwhile, many reports have shown antibiotic resistance rising dramatically. In the present study, we sought to determine the prevalence of microorganisms detected on culture in complicated DFIs in hospitalized patients and their antibiotic sensitivity profiles. Methods. A cross-sectional study was conducted for a period of 24 months from 2012 to 2014 in Nemazee Hospital, Shiraz, Iran. The demographic and clinical features of the patients were obtained. Antimicrobial susceptibility testing to different agents was carried out using the disc diffusion method. Results. During this period, 122 aerobic microorganisms were isolated from DFIs. Among Gram-positive and Gram-negative bacteria, Staphylococcus spp. and E. coli were the most frequent organisms isolated, respectively. Of the isolates, 91% were multidrug while 78% of S. aureus isolates were methicillin resistant. 53% of Gram-negative bacteria were positive for extended-spectrum β-lactamase. Conclusion. Given the involvement of different microorganisms and emergence of multidrug resistant strains, clinicians are advised to consider culture before initiation of empirical therapy. Mojtaba Anvarinejad, Gholamreza Pouladfar, Aziz Japoni, Shahram Bolandparvaz, Zeinab Satiary, Pejman Abbasi, and Jalal Mardaneh Copyright © 2015 Mojtaba Anvarinejad et al. All rights reserved. Endemicity of Acinetobacter calcoaceticus-baumannii Complex in an Intensive Care Unit in Malaysia Sun, 27 Dec 2015 10:16:58 +0000 Introduction. Acinetobacter calcoaceticus-baumannii complex (ACB complex) is a leading opportunistic pathogen in intensive care units (ICUs). Effective control of spread requires understanding of its epidemiological relatedness. This study aims to determine the genetic relatedness and antibiotic susceptibilities of ACB complex in an ICU in Malaysia. Methodology. Pulsed field gel electrophoresis (PFGE), E-test, and disk diffusion were used for isolates characterization. Results. During the study period (December 2011 to June 2012), 1023 patients were admitted to the ICU and 44 ACB complex (blood, , and blind bronchial aspirates, ) were recovered from 38 ICU patients. Six isolates were from non-ICU patients. Of the 44 ICU isolates, 88.6% exhibited multidrug-resistant (MDR) patterns. There was high degree of resistance, with minimum inhibitory concentration90 (MIC90) of >32 μg/mL for carbapenems and ≥256 μg/mL for amikacin, ampicillin/sulbactam, and cefoperazone/sulbactam. Isolates from the main PFGE cluster were highly resistant. There was evidence of dissemination in non-ICU wards. Conclusion. High number of clonally related MDR ACB complex was found. While the ICU is a likely reservoir facilitating transmission, importation from other wards may be important contributor. Early identification of strain relatedness and implementation of infection control measures are necessary to prevent further spread. Amreeta Dhanoa, Ganeswrie Rajasekaram, Soo Sum Lean, Yuet Meng Cheong, and Kwai Lin Thong Copyright © 2015 Amreeta Dhanoa et al. All rights reserved. Restriction Profiling of 23S Microheterogenic Ribosomal Repeats for Detection and Characterizing of E. coli and Their Clonal, Pathogenic, and Phylogroups Tue, 22 Dec 2015 11:22:19 +0000 Correlating ribosomal microheterogenicity with unique restriction profiles can prove to be an efficacious and cost-effective approach compared with sequencing for microbial identification. An attempt to peruse restriction profiling of 23S ribosomal assemblage was ventured; digestion patterns with Bfa I discriminated E. coli from its colony morphovars, while Hae III profiles assisted in establishing distinct clonal groups. Among the gene pool of 399 ribosomal sequences extrapolated from 57 E. coli genomes, varying degree of predominance (I > III > IV > II) of Hae III pattern was observed. This was also corroborated in samples collected from clinical, commensal, and environmental origin. K-12 and its descendants showed type I pattern whereas E. coli-B and its descendants exhibited type IV, both of these patterns being exclusively present in E. coli. A near-possible association between phylogroups and Hae III profiles with presumable correlation between the clonal groups and different pathovars was established. The generic nature, conservation, and barcode gap of 23S rRNA gene make it an ideal choice and substitute to 16S rRNA gene, the most preferred region for molecular diagnostics in bacteria. Parvathi Jayasree Rajagopalan Nair and Sunita Singh Copyright © 2015 Parvathi Jayasree Rajagopalan Nair and Sunita Singh. All rights reserved. Evaluation of Antibiotic Susceptibility of Gram-Positive Anaerobic Cocci Isolated from Cancer Patients of the N. N. Blokhin Russian Cancer Research Center Mon, 21 Dec 2015 08:55:42 +0000 In total, 81 nonduplicate gram-positive anaerobic cocci (GPAC) were involved in this study. The GPAC were isolated from samples collected from cancer patients between 2004 and 2014. Species identification was carried out by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS). The majority of isolates were identified as Finegoldia magna (47%) and Peptoniphilus harei (28%). The susceptibility of six species of GPAC was determined for eight antibiotics according to -test methodology. Furthermore, all isolates were susceptible to imipenem, vancomycin, and linezolid. Susceptibility to penicillin G, amoxicillin/clavulanate, metronidazole, ciprofloxacin, and levofloxacin varied for different species. One Finegoldia magna isolate was multidrug-resistant (i.e., parallel resistance to five antimicrobial agents, including metronidazole, was observed). Two Parvimonas micra isolates were highly resistant to metronidazole (MIC 256 μg/mL) but were sensitive to other tested antibiotics. Irina I. Shilnikova and Natalia V. Dmitrieva Copyright © 2015 Irina I. Shilnikova and Natalia V. Dmitrieva. All rights reserved. Ziziphora clinopodioides Essential Oil and Nisin as Potential Antimicrobial Agents against Escherichia coli O157:H7 in Doogh (Iranian Yoghurt Drink) Sun, 13 Dec 2015 07:29:44 +0000 The aim of the present study was to evaluate the effects of Ziziphora clinopodioides essential oil (0.1 and 0.2%) and nisin (250 and 500 IU/mL) separately and in combination on survival of Escherichia coli O157:H7 inoculated in Doogh (Iranian yoghurt drink) during storage under refrigerated temperature (4 ± 1°C) for 9 days. Viability of Lactobacillus casei at different concentrations of Z. clinopodioides essential oil (0.1 and 0.2%) in Doogh was also examined. The major components were carvacrol (64.22%), thymol (19.22%), -terpinene (4.63%), and -cymene (4.86%). There was no significant difference () between samples treated with nisin and those of untreated samples. Samples treated with both concentrations of the essential oil (0.1 and 0.2%) showed populations of E. coli O157:H7 significantly () lower than those of untreated samples. The essential oil of Z. clinopodioides in combination with nisin had a potential synergistic effect against E. coli O157:H7 in Doogh samples after 5 days. The count of L. casei was not inhibited by different concentrations of the Z. clinopodioides essential oil. It is concluded that the leaf essential oil of Z. clinopodioides in combination with nisin can be applied as alternative antimicrobial agents in Doogh to inhibit the growth of E. coli O157:H7. Yasser Shahbazi Copyright © 2015 Yasser Shahbazi. All rights reserved. Iron Deprivation Affects Drug Susceptibilities of Mycobacteria Targeting Membrane Integrity Tue, 08 Dec 2015 07:28:30 +0000 Multidrug resistance (MDR) acquired by Mycobacterium tuberculosis (MTB) through continuous deployment of antitubercular drugs warrants immediate search for novel targets and mechanisms. The ability of MTB to sense and become accustomed to changes in the host is essential for survival and confers the basis of infection. A crucial condition that MTB must surmount is iron limitation, during the establishment of infection, since iron is required by both bacteria and humans. This study focuses on how iron deprivation affects drug susceptibilities of known anti-TB drugs in Mycobacterium smegmatis, a “surrogate of MTB.” We showed that iron deprivation leads to enhanced potency of most commonly used first line anti-TB drugs that could be reverted upon iron supplementation. We explored that membrane homeostasis is disrupted upon iron deprivation as revealed by enhanced membrane permeability and hypersensitivity to membrane perturbing agent leading to increased passive diffusion of drug and TEM images showing detectable differences in cell envelope thickness. Furthermore, iron seems to be indispensable to sustain genotoxic stress suggesting its possible role in DNA repair machinery. Taken together, we for the first time established a link between cellular iron and drug susceptibility of mycobacteria suggesting iron as novel determinant to combat MDR. Rahul Pal, Saif Hameed, and Zeeshan Fatima Copyright © 2015 Rahul Pal et al. All rights reserved. Evaluation of In Vivo Acaricidal Effect of Soap Containing Essential Oil of Chenopodium ambrosioides Leaves on Rhipicephalus lunulatus in the Western Highland of Cameroon Sun, 06 Dec 2015 08:50:52 +0000 A study on the acaricidal properties of foam soap containing the essential oil of Chenopodium ambrosioides leaves was carried out on Rhipicephalus lunulatus. Four doses (0.03, 0.06, 0.09, and 0.12 µL of essential oil per gram of soap) and a control (soap without essential oil) with four replications for each treatment were used for in vitro trial. Each replication consisted of 10 ticks in a Petri dish with filter paper impregnated with the foam soap on the bottom. Following in vitro trials, three doses (0.06, 0.09, and 0.12 µL/g) and the control in two replications were selected for in vivo test based on mortality rate recorded from the in vitro trial. Each replication was made up of 10 goats naturally infested with ticks. Results show that soap containing essential oil is toxic to R. lunulatus. The in vivo mortality rate in the control on day 8 was 22.69% whereas the highest dose (0.12 µL/g) killed 96.29% of the ticks on day 8. The LD50 of the foam soap containing essential oil was 0.037 and 0.059 µL/g on day 2 in the laboratory and on the farm, respectively. This indicates the potentially high efficiency of this medicated soap on this parasite. Marc K. Kouam, Vincent K. Payne, Emile Miégoué, Fernand Tendonkeng, Jules Lemoufouet, Jean R. Kana, Benoit Boukila, E. Tedonkeng Pamo, and Bertine MNM Copyright © 2015 Marc K. Kouam et al. All rights reserved. Clinical Presentation and Antibiotic Susceptibility of Contact Lens Associated Microbial Keratitis Thu, 03 Dec 2015 08:59:03 +0000 Introduction. In recent years, the number of contact lens wearers has dramatically increased in Iran, particularly in youngsters. The purpose of current study was to assess the clinical presentation and antibiotic susceptibility of contact lens related microbial keratitis in Ahvaz, southwest of Iran. Methodology. A cross-sectional investigation of 26 patients (33 eyes) with contact lens induced corneal ulcers who were admitted to Imam Khomeini Hospital, Ahwaz City, from June 2012 to June 2013 was done. In order to study microbial culture and susceptibility of corneal ulcers, all of them were scraped. Results. Eight samples were reported as sterile. Pseudomonas aeruginosa (80%) in positive cultures was the most widely recognized causative organism isolated. This is followed by Staphylococcus aureus 12% and Enterobacter 8%. The results showed that 84% of the microorganism cases were sensitive to ciprofloxacin, while imipenem, meropenem, and ceftazidime were the second most effective antibiotics (76%). Conclusion. Results of current study show the importance of referring all contact lens wearers with suspected corneal infection to ophthalmologists for more cure. The corneal scraping culture and contact lens solution should be performed to guide antibiotic therapy. Hesam Hedayati, Mahboubeh Ghaderpanah, Seyed Ahmad Rasoulinejad, and Mohammad Montazeri Copyright © 2015 Hesam Hedayati et al. All rights reserved. Comparative Response of the West African Dwarf Goats to Experimental Infections with Red Sokoto and West African Dwarf Goat Isolates of Haemonchus contortus Mon, 30 Nov 2015 11:42:50 +0000 Response of the West African Dwarf (WAD) goats to two different isolates of Haemonchus contortus, the Red Sokoto (RS) goat isolate (RSHc) and the WAD goat isolate (WADHc) (isolated from WAD goats), was studied by experimental infections of 4–6-month-old male WAD goat kids. Group 1 and Group 2 goats were each infected with 4500 infective larvae (L3) of RSHc and WADHc, respectively. Group 3 animals served as uninfected control. Prepatent period (PPP), faecal egg counts (FEC), worm burden (WB), body weight (BW), packed cell volume (PCV), and body condition score (BCS) were determined. WAD goats infected with RSHc isolate and the ones infected with WADHc isolate had mean PPP of and , respectively. Goats infected with WADHc isolate had significantly higher FEC () and WB (). BW were significantly higher () both in the controls and in Group 2 goats infected with WADHc isolate than in Group 1 goats infected with the RSHc isolate. BCS of animals in both infected groups dropped significantly (). There was a significant drop in PCV () of both infected groups in comparison. Both isolates of H. contortus were pathogenic to the host. Lucas Atehmengo Ngongeh and Amaechi Onyeabor Copyright © 2015 Lucas Atehmengo Ngongeh and Amaechi Onyeabor. All rights reserved. No Polymorphism in Plasmodium falciparum K13 Propeller Gene in Clinical Isolates from Kolkata, India Wed, 25 Nov 2015 11:31:14 +0000 Molecular markers associated with artemisinin resistance in Plasmodium falciparum are yet to be well defined. Recent studies showed that polymorphisms in K13 gene are associated with artemisinin resistance. The present study was designed to know the pattern of polymorphisms in propeller region of K13 gene among the clinical isolates collected from urban Kolkata after five years of ACT implementation. We collected 59 clinical isolates from urban Kolkata and sequenced propeller region of K13 gene in 51 isolates successfully. We did not find any mutation in any isolate. All patients responded to the ACT, a combination of artesunate + sulphadoxine-pyrimethamine. The drug regimen is still effective in the study area and there is no sign of emergence of resistance against artemisinin as evidenced by wild genotype of K13 gene in all isolates studied. Moytrey Chatterjee, Swagata Ganguly, Pabitra Saha, Biswabandhu Bankura, Nandita Basu, Madhusudan Das, Subhasish K. Guha, and Ardhendu K. Maji Copyright © 2015 Moytrey Chatterjee et al. All rights reserved.