Journal of Pathogens The latest articles from Hindawi Publishing Corporation © 2015 , Hindawi Publishing Corporation . 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. Prevalence of Plasmid-Mediated Quinolone Resistance Genes among Extended-Spectrum β-Lactamase-Producing Klebsiella pneumoniae Human Isolates in Iran Wed, 04 Nov 2015 12:06:57 +0000 The purpose of this study was to determine the prevalence and molecular characterization of plasmid-mediated quinolone resistance (PMQR) genes (qnrA, qnrB, qnrS, aac(6′)-Ib-cr, and qepA) among ESBL-producing Klebsiella pneumoniae isolates in Kashan, Iran. A total of 185 K. pneumoniae isolates were tested for quinolone resistance and ESBL-producing using the disk diffusion method and double disk synergy (DDST) confirmatory test. ESBL-producing strains were further evaluated for the genes. The PCR method was used to show presence of plasmid-mediated quinolone resistance genes and the purified PCR products were sequenced. Eighty-seven ESBL-producing strains were identified by DDST confirmatory test and majority (70, 80.5%) of which carried genes including CTX-M-1 (60%), CTX-M-2 (42.9%), and CTX-M-9 (34.3%). Seventy-seven ESBL-producing K. pneumoniae isolates harbored PMQR genes, which mostly consisted of aac(6′)-Ib-cr (70.1%) and qnrB (46.0%), followed by qnrS (5.7%). Among the 77 PMQR-positive isolates, 27 (35.1%) and 1 (1.3%) carried 2 and 3 different PMQR genes, respectively. However, qnrA and qepA were not found in any isolate. Our results highlight high ESBL occurrence with CTX-M type and high frequency of plasmid-mediated quinolone resistance genes among ESBL-producing K. pneumoniae isolates in Kashan. Ehsaneh Shams, Farzaneh Firoozeh, Rezvan Moniri, and Mohammad Zibaei Copyright © 2015 Ehsaneh Shams et al. All rights reserved. General Overview on Nontuberculous Mycobacteria, Biofilms, and Human Infection Wed, 04 Nov 2015 09:33:57 +0000 Nontuberculous mycobacteria (NTM) are emergent pathogens whose importance in human health has been growing. After being regarded mainly as etiological agents of opportunist infections in HIV patients, they have also been recognized as etiological agents of several infections on immune-competent individuals and healthcare-associated infections. The environmental nature of NTM and their ability to assemble biofilms on different surfaces play a key role in their pathogenesis. Here, we review the clinical manifestations attributed to NTM giving particular importance to the role played by biofilm assembly. Sonia Faria, Ines Joao, and Luisa Jordao Copyright © 2015 Sonia Faria et al. All rights reserved. Human Papillomavirus Infection, Infertility, and Assisted Reproductive Outcomes Sun, 01 Nov 2015 09:15:59 +0000 The human papillomavirus (HPV) is a sexually transmitted infection common among men and women across all geographic and socioeconomic subgroups worldwide. Recent evidence suggests that HPV infection may affect fertility and alter the efficacy of assisted reproductive technologies. In men, HPV infection can affect sperm parameters, specifically motility. HPV-infected sperm can transmit viral DNA to oocytes, which may be expressed in the developing blastocyst. HPV can increase trophoblastic apoptosis and reduce the endometrial implantation of trophoblastic cells, thus increasing the theoretical risk of miscarriage. Vertical transmission of HPV during pregnancy may be involved in the pathophysiology of preterm rupture of membranes and spontaneous preterm birth. In patients undergoing intrauterine insemination for idiopathic infertility, HPV infection confers a lower pregnancy rate. In contrast, the evidence regarding any detrimental impact of HPV infection on IVF outcomes is inconclusive. It has been suggested that vaccination could potentially counter HPV-related sperm impairment, trophoblastic apoptosis, and spontaneous miscarriages; however, these conclusions are based on in vitro studies rather than large-scale epidemiological studies. Improvement in the understanding of HPV sperm infection mechanisms and HPV transmission into the oocyte and developing blastocyst may help explain idiopathic causes of infertility and miscarriage. Nigel Pereira, Katherine M. Kucharczyk, Jaclyn L. Estes, Rachel S. Gerber, Jovana P. Lekovich, Rony T. Elias, and Steven D. Spandorfer Copyright © 2015 Nigel Pereira et al. All rights reserved. Protective Effects of Tinospora crispa Stem Extract on Renal Damage and Hemolysis during Plasmodium berghei Infection in Mice Tue, 27 Oct 2015 07:23:37 +0000 Renal damage and hemolysis induced by malaria are associated with mortality in adult patients. It has been speculated that oxidative stress condition induced by malaria infection is involved in its pathology. Thus, we aimed to investigate the protective effects of Tinospora crispa stem extract on renal damage and hemolysis during Plasmodium berghei infection. T. crispa stem extract was prepared using hot water method and used for oral treatment in mice. Groups of ICR mice were infected with parasitized erythrocytes of P. berghei ANKA by intraperitoneal injection and given the extracts (500, 1000, and 2000 mg/kg) twice a day for 4 consecutive days. To assess renal damage and hemolysis, blood urea nitrogen (BUN), creatinine, and hematocrit (%Hct) levels were then evaluated, respectively. Malaria infection resulted in renal damage and hemolysis as indicated by increasing of BUN and creatinine and decreasing of %Hct, respectively. However, protective effects on renal damage and hemolysis were observed in infected mice treated with these extracts at doses of 1000 and 2000 mg/kg. In conclusion, T. crispa stem extract exerted protective effects on renal damage and hemolysis induced by malaria infection. This plant may work as potential source in the development of variety of herbal formulations for malarial treatment. Narain Nutham, Sakuna Sakulmettatham, Suwit Klongthalay, Palatip Chutoam, and Voravuth Somsak Copyright © 2015 Narain Nutham et al. All rights reserved. An Additive Effect of Oral N-Acetyl Cysteine on Eradication of Helicobacter pylori Wed, 02 Sep 2015 09:17:43 +0000 Background. Helicobacter pylori is highly adapted to the gastric environment where it lives within or beneath the gastric mucous layer. The aim of this study was to evaluate whether the addition of N-acetyl cysteine to the treatment regimen of H. pylori infection would affect eradication rates of the disease. Methods. A total of 79 H. pylori positive patients were randomized to two therapeutic groups. Both groups received a 14-day course of three-drug regimen including amoxicillin/clarithromycin/omeprazole. Experimental group (38 subjects) received NAC, and control group (41 subjects) received placebo, besides three-drug regimen. H. pylori eradication was evaluated by urea breath test at least 4 weeks after the cessation of therapy. Results. The rate of H. pylori eradication was 72.9% and 60.9% in experimental and control groups, respectively (). By logistic regression modeling, female gender (OR 3.68, 95% CI: 1.06–5.79; ) and treatment including NAC (OR 1.88, 95% CI: 0.68–3.15; ) were independent factors associated with H. pylori eradication. Conclusion. The results of the present study show that NAC has an additive effect on the eradication rates of H. pylori obtained with three-drug regimen and appears to be a promising means of eradicating H. pylori infection. Seyed Mohammad-Taghi Hamidian, Najmeh-sadat Aletaha, Reza Taslimi, and Mohammad Montazeri Copyright © 2015 Seyed Mohammad-Taghi Hamidian et al. All rights reserved. Prevalence of Virulence Factors and Drug Resistance in Clinical Isolates of Enterococci: A Study from North India Sun, 23 Aug 2015 11:24:33 +0000 Along with emergence of multidrug resistance, presence of several virulence factors in enterococci is an emerging concept. This study was undertaken to determine the prevalence of various virulence factors phenotypically and genotypically in enterococci and study their association with multidrug resistance. A total of 310 enterococcal isolates were studied, comprising 155 E. faecium and 155 E. faecalis. Antimicrobial susceptibility testing was done by disc diffusion and agar dilution method. Hemolysin, gelatinase, biofilm production, and haemagglutination were detected phenotypically and presence of virulence genes, namely, asa1, gelE, cylA, esp, and hyl, was detected by multiplex PCR. Of the total, 47.41% isolates were high level gentamicin resistant (HLGRE) and 7.09% were vancomycin resistant (VRE). All the virulence traits studied were found in varying proportions, with majority in E. faecalis (). Strong biofilm producers possessed either asa1 or gelE gene. gelE silent gene was detected in 41.37% (12/29). However, increase in resistance was associated with significant decrease in expression or acquisition of virulence genes. Further, acquisition of vancomycin resistance was the significant factor responsible for the loss of virulence traits. Though it is presumed that increased drug resistance correlates with increased virulence, acquisition of vancomycin resistance might be responsible for reduced expression of virulence traits to meet the “biological cost” relating to VRE. Tuhina Banerjee and Shampa Anupurba Copyright © 2015 Tuhina Banerjee and Shampa Anupurba. All rights reserved. Chemical Composition and In Vitro Antibacterial Activity of Mentha spicata Essential Oil against Common Food-Borne Pathogenic Bacteria Sun, 16 Aug 2015 12:03:14 +0000 The aim of the present study was to investigate chemical composition and antibacterial activity of essential oil from the leaf of Mentha spicata plant against common food-borne pathogenic bacteria (Staphylococcus aureus, Bacillus subtilis, Bacillus cereus, Listeria monocytogenes, Salmonella typhimurium, and Escherichia coli O157:H7). Chemical composition of the essential oil was identified by gas chromatography coupled with mass spectrometer detector (GC-MS). The antibacterial activity of the essential oil was evaluated by broth microdilution method and agar disk diffusion assay. According to the result of GC-MS analysis, 18 components were identified, accounting for 99.89% of the whole essential oil. The main components were carvone (78.76%), limonene (11.50%), β-bourbonene (11.23%), cis-dihydrocarveol (1.43%), trans-caryophyllene (1.04%), menthone (1.01%), menthol (1%), and terpinen-4-ol (0.99). The essential oil exhibited moderate level of antibacterial activity against all test microorganisms. In general, Gram-positive bacteria were more susceptible to M. spicata essential oil than Gram-negative bacteria. L. monocytogenes was the most sensitive of the microorganisms to the antibacterial activity of M. spicata essential oil (inhibition zone = 22 mm and MIC and MBC = 2.5 µL/mL). Based on our results, the essential oil of M. spicata plant collected from Kermanshah province, west of Iran, has a potential to be applied as antibacterial agent. Yasser Shahbazi Copyright © 2015 Yasser Shahbazi. All rights reserved. Chlamydia trachomatis and Genital Mycoplasmas: Pathogens with an Impact on Human Reproductive Health Wed, 31 Dec 2014 00:10:19 +0000 The most prevalent, curable sexually important diseases are those caused by Chlamydia trachomatis (C. trachomatis) and genital mycoplasmas. An important characteristic of these infections is their ability to cause long-term sequels in upper genital tract, thus potentially affecting the reproductive health in both sexes. Pelvic inflammatory disease (PID), tubal factor infertility (TFI), and ectopic pregnancy (EP) are well documented complications of C. trachomatis infection in women. The role of genital mycoplasmas in development of PID, TFI, and EP requires further evaluation, but growing evidence supports a significant role for these in the pathogenesis of chorioamnionitis, premature membrane rupture, and preterm labor in pregnant woman. Both C. trachomatis and genital mycoplasmas can affect the quality of sperm and possibly influence the fertility of men. For the purpose of this paper, basic, epidemiologic, clinical, therapeutic, and public health issue of these infections were reviewed and discussed, focusing on their impact on human reproductive health. Sunčanica Ljubin-Sternak and Tomislav Meštrović Copyright © 2014 Sunčanica Ljubin-Sternak and Tomislav Meštrović. All rights reserved. Clostridium difficile with Moxifloxacin/Clindamycin Resistance in Vegetables in Ohio, USA, and Prevalence Meta-Analysis Sun, 14 Dec 2014 00:10:12 +0000 We (i) determined the prevalence of Clostridium difficile and their antimicrobial resistance to six antimicrobial classes, in a variety of fresh vegetables sold in retail in Ohio, USA, and (ii) conducted cumulative meta-analysis of reported prevalence in vegetables since the 1990s. Six antimicrobial classes were tested for their relevance as risk factors for C. difficile infections (CDIs) (clindamycin, moxifloxacin) or their clinical priority as exhaustive therapeutic options (metronidazole, vancomycin, linezolid, and tigecycline). By using an enrichment protocol we isolated C. difficile from three of 125 vegetable products (2.4%). All isolates were toxigenic, and originated from 4.6% of 65 vegetables cultivated above the ground (; outer leaves of iceberg lettuce, green pepper, and eggplant). Root vegetables yielded no C. difficile. The C. difficile isolates belonged to two PCR ribotypes, one with an unusual antimicrobial resistance for moxifloxacin and clindamycin (lettuce and pepper; 027-like, A+B+CDT+; tcdC 18 bp deletion); the other PCR ribotype (eggplant, A+B+ CDT−; classic tcdC) was susceptible to all antimicrobials. Results of the cumulative weighted meta-analysis (6 studies) indicate that the prevalence of C. difficile in vegetables is 2.1% and homogeneous since the first report in 1996 (2.4%). The present study is the first report of the isolation of C. difficile from retail vegetables in the USA. Of public health relevance, antimicrobial resistance to moxifloxacin/clindamycin (a bacterial-associated risk factor for severe CDIs) was identified on the surface of vegetables that are consumed raw. Alex Rodriguez-Palacios, Sanja Ilic, and Jeffrey T. LeJeune Copyright © 2014 Alex Rodriguez-Palacios et al. All rights reserved. Type I Diabetes Mellitus: Genetic Factors and Presumptive Enteroviral Etiology or Protection Wed, 10 Dec 2014 06:33:31 +0000 We review type 1 diabetes and host genetic components, as well as epigenetics and viruses associated with type 1 diabetes, with added emphasis on the enteroviruses, which are often associated with triggering the disease. Genus Enterovirus is classified into twelve species of which seven (Enterovirus A, Enterovirus B, Enterovirus C, and Enterovirus D and Rhinovirus A, Rhinovirus B, and Rhinovirus C) are human pathogens. These viruses are transmitted mainly by the fecal-oral route; they may also spread via the nasopharyngeal route. Enterovirus infections are highly prevalent, but these infections are usually subclinical or cause a mild flu-like illness. However, infections caused by enteroviruses can sometimes be serious, with manifestations of meningoencephalitis, paralysis, myocarditis, and in neonates a fulminant sepsis-like syndrome. These viruses are often implicated in chronic (inflammatory) diseases as chronic myocarditis, chronic pancreatitis, and type 1 diabetes. In this review we discuss the currently suggested mechanisms involved in the viral induction of type 1 diabetes. We recapitulate current basic knowledge and definitions. Jana Precechtelova, Maria Borsanyiova, Sona Sarmirova, and Shubhada Bopegamage Copyright © 2014 Jana Precechtelova et al. All rights reserved. Sesamol: A Natural Phenolic Compound with Promising Anticandidal Potential Tue, 09 Dec 2014 12:37:53 +0000 We investigated the antifungal effects of sesamol (Ses), a natural phenolic compound, and exemplified that it could be mediated through disruption of calcineurin signaling pathway in C. albicans, a human fungal pathogen. The repertoire of antifungal activity not only was limited to C. albicans and its six clinical isolates tested but also was against non-albicans species of Candida. Interestingly, the antifungal effect of Ses affects neither the MDR efflux transporter activity nor passive diffusion of drug. We found that C. albicans treated with Ses copies the phenotype displayed by cells having defect in calcineurin signaling leading to sensitivity against alkaline pH, ionic, membrane, salinity, endoplasmic reticulum, and serum stresses but remained resistant to thermal stress. Furthermore, the ergosterol levels were significantly decreased by 63% confirming membrane perturbations in response to Ses as also visualized through transmission electron micrographs. Despite the fact that Ses treatment mimics the phenotype of compromised calcineurin signaling, it was independent of cell wall integrity pathway as revealed by spot assays and the scanning electron micrographs. Taken together, the data procured from this study clearly ascertains that Ses is an effectual antifungal agent that could be competently employed in treating Candida infections. Moiz A. Ansari, Zeeshan Fatima, and Saif Hameed Copyright © 2014 Moiz A. Ansari et al. All rights reserved. Antihemolytic Activities of Green Tea, Safflower, and Mulberry Extracts during Plasmodium berghei Infection in Mice Tue, 18 Nov 2014 09:09:34 +0000 Malaria-associated hemolysis is associated with mortality in adult patients. It has been speculated that oxidative stress and inflammation induced by malaria parasite are involved in its pathophysiology. Hence, we aimed to investigate the antihemolytic effect of green tea, safflower, and mulberry extracts against Plasmodium berghei infection. Aqueous crude extracts of these plants were prepared using hot water method and used for oral treatment in mice. Groups of ICR mice were infected with 6 × 106 infected red blood cells of P. berghei ANKA by intraperitoneal injection and given the extracts (500, 1500, and 3000 mg/kg) twice a day for 4 consecutive days. To assess hemolysis, hematocrit levels were then evaluated. Malaria infection resulted in hemolysis. However, antihemolytic effects were observed in infected mice treated with these extracts at dose-dependent manners. In conclusion, aqueous crude extracts of green tea, safflower, and mulberry exerted antihemolysis induced by malaria infection. These plants may work as potential source in the development of variety of herbal formulations for malarial treatment. Suthin Audomkasok, Waraporn Singpha, Sukanya Chachiyo, and Voravuth Somsak Copyright © 2014 Suthin Audomkasok et al. All rights reserved. Effective Concentration and Detection of Cryptosporidium, Giardia, and the Microsporidia from Environmental Matrices Wed, 10 Sep 2014 08:17:45 +0000 Cryptosporidium spp., Giardia spp., and members of Microsporidia are enteropathogenic parasites of humans and animals, producing asymptomatic to severe intestinal infections. To circumvent various impediments associated with current detection methods, we tested a method providing multistage purification and separation in a single, confined step. Standard real-time PCR was used as a detection method. Samples spiked with C. parvum and G. intestinalis were split for comparison to standard Method 1623. Results were equivalent to immunomagnetic procedures for Cryptosporidium, and Giardia. Overall percent recovery for Cryptosporidium with Method 1623 averaged 26.89% (std 21.44%; min = 0%; max = 73%) and was similar but less variable for qPCR method at an estimated average of 27.67 (std 17.65%; min = 5%; max = 63%). For Giardia, Method 1623 had an overall average recovery of 27.11% (std 17.98%; min = 1%; max = 58%), while multistage purification and qPCR had an estimated lower overall recovery at 18.58% (std 13.95%; min = 0%; max = 35%). Microsporidia were also readily detected with an estimated recovery of 46.81% overall (std 17.66%; min = 18%; max = 70%) for E. intestinalis and 38.90% (std 14.36%; min = 13%; max = 62%) for E. bieneusi. Joseph A. Moss, John Gordy, and Richard A. Snyder Copyright © 2014 Joseph A. Moss et al. All rights reserved. Prevalence and Diversity among Anaplasma phagocytophilum Strains Originating from Ixodes ricinus Ticks from Northwest Norway Sun, 24 Aug 2014 05:45:39 +0000 The tick-borne pathogen Anaplasma phagocytophilum causes great concern for livestock farmers. Tick-borne fever is a widespread disease in Norway, and antibodies have been produced amongst sheep, roe deer, red deer, and moose. The main vector Ixodes ricinus is found along the Norwegian coastline as far north as the Arctic Circle. A total number of 1804 I. ricinus ticks were collected and the prevalence of the pathogen was determined by species-specific qPCR. The overall infection rate varied from 2.83% to 3.32%, but there were no significant differences () in the overall infection rate in 2010, 2011, or 2012. A multilocus sequencing analysis was performed to further characterise the isolates. The genotyping of 27 strains resulted in classification into 19 different sequences types (ST), none of which was found in the MLST database. The nucleotide diversity was for every locus <0.01, and the number of SNPs was between 1 and 2.8 per 100 bp. The majority of SNPs were synonymous. A goeBURST analysis demonstrated that the strains from northwest Norway cluster together with other Norwegian strains in the MLST database and the strains that are included in this study constitute clonal complexes (CC) 9, 10, and 11 in addition to the singleton. Ann-Kristin Tveten Copyright © 2014 Ann-Kristin Tveten. All rights reserved. Concomitant Colonization of Helicobacter pylori in Dental Plaque and Gastric Biopsy Wed, 09 Jul 2014 10:49:34 +0000 Frequently reported H. pylori antimicrobial therapy failures suggest that there might be a different niche where the bacteria can stay safe. Current study aims to examine potential role of oral colonization of H. pylori to feed reinfection after primary therapy. However, patients who were admitted to the gastroscopy section were chosen and gastric biopsy and dental plaque specimens were collected. Molecular and biochemical tests were applied to confirm H. pylori identity in different colonization niches. Results showed that 88.8% of dyspeptic patients had epigastric pains with nocturnal awakening when they were hungry (). All patients who received therapy already were again H. pylori positive while they are still carrying H. pylori in dental plaque (). Moreover, H. pylori infection was sought in 100% of gastric biopsy’s dyspeptic patients who had ulcerated esophagitis and erosive duodenitis and who were H. pylori positive, and 75% of dyspeptic patients with duodenum deformity had this bacterium in gastric biopsies (). Present study showed that only successful eradication of gastric H. pylori cannot guarantee prevention of reinfection. Conclusively, a new strategy which indicates concomitant eradication in oral and gastric colonization can result in clearance of H. pylori infection. Amin Talebi Bezmin Abadi, Ashraf Mohabati Mobarez, Omid Teymournejad, and Mona Karbalaei Copyright © 2014 Amin Talebi Bezmin Abadi et al. All rights reserved. Antifungal Susceptibility Patterns, In Vitro Production of Virulence Factors, and Evaluation of Diagnostic Modalities for the Speciation of Pathogenic Candida from Blood Stream Infections and Vulvovaginal Candidiasis Mon, 07 Jul 2014 11:49:27 +0000 Candida spp. have emerged as successful pathogens in both invasive and mucosal infections. Varied virulence factors and growing resistance to antifungal agents have contributed to their pathogenicity. We studied diagnostic accuracy of HiCrome Candida Differential Agar and Vitek 2 Compact system for identification of Candida spp. in comparison with species-specific PCR on 110 clinical isolates of Candida from blood stream infections (54, 49%) and vulvovaginal candidiasis (56, 51%). C. albicans (61%) was the leading pathogen in VVC, while C. tropicalis (46%) was prominent among BSIs. HiCrome Agar and Vitek 2 Compact had good measures of agreement (κ) 0.826 and 0.895, respectively, in comparison with PCR. We also tested these isolates for in vitro production of proteinase, esterase, phospholipases, and biofilms. Proteinase production was more among invasive isolates (), while phospholipase production was more among noninvasive isolates (). There was an overall increase in the production of virulence factors among non-albicans Candida. Identification of clinical isolates of Candida up to species level either by chromogenic agar or by Vitek 2 Compact system should be routinely done to choose appropriate therapy. Chaitanya Tellapragada, Vandana Kalwaje Eshwara, Ruqaiyah Johar, Tushar Shaw, Nidhi Malik, Parvati V. Bhat, Asha Kamath, and Chiranjay Mukhopadhyay Copyright © 2014 Chaitanya Tellapragada et al. All rights reserved. Isolation of Environmental Bacteria from Surface and Drinking Water in Mafikeng, South Africa, and Characterization Using Their Antibiotic Resistance Profiles Sun, 06 Jul 2014 00:00:00 +0000 The aim of this study was to isolate and identify environmental bacteria from various raw water sources as well as the drinking water distributions system in Mafikeng, South Africa, and to determine their antibiotic resistance profiles. Water samples from five different sites (raw and drinking water) were analysed for the presence of faecal indicator bacteria as well as Aeromonas and Pseudomonas species. Faecal and total coliforms were detected in summer in the treated water samples from the Modimola dam and in the mixed water samples, with Pseudomonas spp. being the most prevalent organism. The most prevalent multiple antibiotic resistance phenotype observed was KF-AP-C-E-OT-K-TM-A. All organisms tested were resistant to erythromycin, trimethoprim, and amoxicillin. All isolates were susceptible to ciprofloxacin and faecal coliforms and Pseudomonas spp. to neomycin and streptomycin. Cluster analysis based on inhibition zone diameter data suggests that the isolates had similar chemical exposure histories. Isolates were identified using gyrB, toxA, ecfX, aerA, and hylH gene fragments and gyrB, ecfX, and hylH fragments were amplified. These results demonstrate that (i) the drinking water from Mafikeng contains various bacterial species and at times faecal and total coliforms. (ii) The various bacteria are resistant to various classes of antibiotics. Suma George Mulamattathil, Carlos Bezuidenhout, Moses Mbewe, and Collins Njie Ateba Copyright © 2014 Suma George Mulamattathil et al. All rights reserved. Prevalence of Multiple Antibiotic Resistant Infections in Diabetic versus Nondiabetic Wounds Wed, 25 Jun 2014 07:00:56 +0000 Diabetes mellitus (DM) affects 23.6 million people in the USA and approximately 20–25% of diabetic patients will develop foot ulceration during the course of their disease. Up to a quarter of these patients will develop infections that will necessitate amputation. Although many studies report that the rates of antibiotic resistant infections have increased dramatically in the DM population over the last decade, to our knowledge there have been no reports directly comparing the rates of antibiotic resistant infections in DM versus non-DM wounds. We performed a retrospective study comparing the wound infections of 41 DM patients to those of 74 non-DM patients to test the hypothesis that infections with multidrug resistant organisms (MDRO) were more prevalent in the DM population. We found that 63.4% of DM and 50% of non-DM patients had MDRO infections, which was not statistically different. However, 61% of the DM patients had Pseudomonas infections compared to only 18.9% of non-DM patients. Furthermore, DM patients had significantly more coinfections with both Pseudomonas and Staphylococcus aureus. Though our initial hypothesis was incorrect, we demonstrated a significant correlation between Pseudomonas and Pseudomonas/S. aureus coinfections within DM wounds. Urvish Trivedi, Shamini Parameswaran, Andrew Armstrong, Diana Burgueno-Vega, John Griswold, Sharmila Dissanaike, and Kendra P. Rumbaugh Copyright © 2014 Urvish Trivedi et al. All rights reserved. Antimicrobial Susceptibility of Escherichia coli Isolated from Fresh-Marketed Nile Tilapia (Oreochromis niloticus) Mon, 07 Apr 2014 09:51:49 +0000 The contamination of seafood by bacteria of fecal origin, especially Escherichia coli, is a widely documented sanitary problem. The objective of the present study was to isolate E. coli strains from the gills, muscle, and body surface of farmed Nile tilapias (Oreochromis niloticus) fresh-marketed in supermarkets in Fortaleza (Ceará, Brazil), to determine their susceptibility to antibiotics of different families (amikacin, gentamicin, imipenem, cephalothin, cefotaxime, ciprofloxacin, aztreonam, ampicillin, nalidixic acid, tetracycline, and sulfametoxazol-trimetoprim), and to determine the nature of resistance by plasmid curing. Forty-four strains (body surface = 25, gills = 15, muscle = 4) were isolated, all of which were susceptible to amikacin, aztreonam, cefotaxime, ciprofloxacin, gentamicin, and imipenem. Gill and body surface samples yielded 11 isolates resistant to ampicillin, tetracycline, and sulfametoxazol-trimetoprim, 4 of which of plasmidial nature. The multiple antibiotic resistance index was higher for strains isolated from body surface than from gills. The overall high antibiotic susceptibility of E. coli strains isolated from fresh-marketed tilapia was satisfactory, although the occasional finding of plasmidial resistance points to the need for close microbiological surveillance of the farming, handling, and marketing conditions of aquaculture products. Rafael dos Santos Rocha, Lana Oliveira Leite, Oscarina Viana de Sousa, and Regine Helena Silva dos Fernandes Vieira Copyright © 2014 Rafael dos Santos Rocha et al. All rights reserved. Detection of Legionella spp. from Domestic Water in the Prefecture of Arta, Greece Wed, 12 Mar 2014 07:36:38 +0000 The aim of this research was the isolation of Legionella spp. from domestic water supply networks in the Prefecture of Arta. A total of 100 water samples, from 25 houses, were collected. Half of the samples concerned the cold water and half the hot water supply. Purpose was to detect colonization of the water networks with Legionella spp. >500 cfu/L by using the method of filtration (ISO 11731). Out of 100 samples, 6 samples from 3 houses were positive for Legionella spp. Legionella pneumophila serogroup 2–14 was isolated in 5 of 6 samples, whereas in the sixth sample Legionella anisa was identified. Only three of the samples had residual chloride over 0.2 mg/L, rate which is necessary for potable water, according to the Greek hygienic practice. Concerning the temperature of hot water, the mean temperature of the negative for Legionella samples was higher compared to the mean temperature of the positive for Legionella samples (49.9°C versus 45.5°C). It is estimated that there is risk of infection through the use of showers. The low concentration of chloride and the temperature, which was found within the limits favorable to developing Legionella spp. (20–45°C), provide fertile ground for proliferation of the bacteria. Dimitra Dimitriadi and Emmanuel Velonakis Copyright © 2014 Dimitra Dimitriadi and Emmanuel Velonakis. All rights reserved. Correlation between PFGE Groups and mrp/epf/sly Genotypes of Human Streptococcus suis Serotype 2 in Northern Thailand Thu, 06 Mar 2014 09:01:46 +0000 Streptococcus suis infection is a severe zoonotic disease commonly found in Northern Thailand where people often consume raw pork and/or pig’s blood. The most frequent clinical presentations are meningitis, sepsis, and endocarditis with higher rate of mortality and hearing loss sequelae. To clarify the correlation between pulsed-field gel electrophoresis (PFGE) groups and mrp/epf/sly genotypes of S. suis serotype 2, 62 patient and 4 healthy pig isolates from Northern Thailand were studied. By PFGE analysis, at 66% homology, most human isolates (69.4%) and 1 pig isolate were in group A, whereas 14.5% of human isolates and 3 out of 4 pig isolates were in group D. According to mrp/epf/sly genotypes, 80.6% of human isolates were identified in mrp+epf−sly− and only 12.9% were in mrp−epf−sly+ genotypes; in contrast, 1 and 3 pig isolates were detected in these two genotypes, respectively. Interestingly, all isolates of S. suis serotype 2 classified in PFGE groups A, B, and E were set in mrp+epf−sly− genotypes. These data show a close correlation between PFGE groups and mrp/epf/sly genotypes of human S. suis serotype 2. Prasit Tharavichitkul, Kanreuthai Wongsawan, Naoki Takenami, Sumalee Pruksakorn, Achara Fongcom, Marcelo Gottschalk, Banyong Khanthawa, Volaluk Supajatura, and Shinji Takai Copyright © 2014 Prasit Tharavichitkul et al. All rights reserved. Tailoring the Immune Response via Customization of Pathogen Gene Expression Tue, 25 Feb 2014 13:02:41 +0000 The majority of studies focused on the construction and reengineering of bacterial pathogens have mainly relied on the knocking out of virulence factors or deletion/mutation of amino acid residues to then observe the microbe’s phenotype and the resulting effect on the host immune response. These knockout bacterial strains have also been proposed as vaccines to combat bacterial disease. Theoretically, knockout strains would be unable to cause disease since their virulence factors have been removed, yet they could induce a protective memory response. While knockout strains have been valuable tools to discern the role of virulence factors in host immunity and bacterial pathogenesis, they have been unable to yield clinically relevant vaccines. The advent of synthetic biology and enhanced user-directed gene customization has altered this binary process of knockout, followed by observation. Recent studies have shown that a researcher can now tailor and customize a given microbe’s gene expression to produce a desired immune response. In this commentary, we highlight these studies as a new avenue for controlling the inflammatory response as well as vaccine development. Lisa M. Runco, Charles B. Stauft, and J. Robert Coleman Copyright © 2014 Lisa M. Runco et al. All rights reserved. High Prevalence of Helicobacter pylori hopQ II Genotype Isolated from Iranian Patients with Gastroduodenal Disorders Mon, 10 Feb 2014 07:13:53 +0000 Helicobacter pylori plays an important role in the pathogenesis of chronic gastritis, peptic ulceration, and noncardia gastric cancer. Several putative virulence factors for H. pylori have been identified including vacA, babA, and iceA. HopQ is one of the outer membrane proteins involved in bacterial adherence to gastric mucosa and has been suggested to also play a role in the virulence of H. pylori. Due to the substantial geographic differences in the prevalence of H. pylori virulence factors reported, the main purpose of the current study was to investigate the association between different H. pylori virulence hopQ alleles (types I and II) and patients with gastroduodenal disorders. The presence of H. pylori and hopQ alleles in gastric biopsy specimens was identified by specific PCR assays. H. pylori type II hopQ was found to be significantly associated with gastric cancer patients (odds ratio: 3.47, 95% CI: 1.56–5.89). Information about the prevalence of H. pylori hopQ type II can be used for determining the high-risk diseases type which is actually colonized by H. pylori hopQ type II positive strains. The presence of H. pylori hopQ type II should be investigated in different geographical regions as confirmatory findings may provide a definite biomarker attributed to the pathogenesis of certain severe digestive diseases. Amin Talebi Bezmin Abadi and Ashraf Mohabbati Mobarez Copyright © 2014 Amin Talebi Bezmin Abadi and Ashraf Mohabbati Mobarez. All rights reserved. Pathogenicity, Ovicidal Action, and Median Lethal Concentrations (LC50) of Entomopathogenic Fungi against Exotic Spiralling Whitefly, Aleurodicus dispersus Russell Mon, 23 Dec 2013 09:34:13 +0000 Biological control using entomopathogenic fungi could be a promising alternative to chemical control. Entomopathogenic fungi, Beauveria bassiana (Balsamo) Vuillemin, Metarhizium anisopliae (Metschnikoff) Sorokin, Lecanicillium lecanii (Zimmerm.) Zare and Gams, and Paecilomyces fumosoroseus (Wize) Brown and Smith, were tested for their pathogenicity, ovicidal effect, and median lethal concentrations (LC50) against exotic spiralling whitefly, Aleurodicus dispersus Russell. The applications were made at the rate of 2 × 109 conidia mL−1 for evaluating the pathogenicity and ovicidal effect of entomopathogenic fungi against A. dispersus. The results of pathogenicity test showed that P. fumosoroseus (P1 strain) was highly pathogenic to A. dispersus recording 100% mortality at 15 days after treatment (DAT). M. anisopliae (M2 strain) had more ovicidal effect causing 37.3% egg mortality at 8 DAT. However, L. lecanii (L1 strain) caused minimum egg hatchability (23.2%) at 10 DAT as compared to control (92.6%). The lowest LC50 produced by P. fumosoroseus (P1 strain) as 8.189 × 107 conidia mL−1 indicated higher virulence against A. dispersus. Hence, there is potential for use of entomopathogenic fungi in the field conditions as an alternate control method in combating the insect pests and other arthropod pests since they are considered natural mortality agents and are environmentally safe. Boopathi Thangavel, Karuppuchamy Palaniappan, Kalyanasundaram Manickavasagam Pillai, Mohankumar Subbarayalu, and Ravi Madhaiyan Copyright © 2013 Boopathi Thangavel et al. All rights reserved. Antibiotic Resistance of Salmonella spp. Isolated from Shrimp Farming Freshwater Environment in Northeast Region of Brazil Sun, 22 Dec 2013 14:22:14 +0000 This study investigated the presence and antibiotic resistance of Salmonella spp. in a shrimp farming environment in Northeast Region of Brazil. Samples of water and sediments from two farms rearing freshwater-acclimated Litopenaeus vannamei were examined for the presence of Salmonella. Afterwards, Salmonella isolates were serotyped, the antimicrobial resistance was determined by a disk diffusion method, and the plasmid curing was performed for resistant isolates. A total of 30 (16.12%) of the 186 isolates were confirmed to be Salmonella spp., belonging to five serovars: S. serovar Saintpaul, S. serovar Infantis, S. serovar Panama, S. serovar Madelia, and S. serovar Braenderup, along with 2 subspecies: S. enterica serovar houtenae and S. enterica serovar enterica. About twenty-three percent of the isolates were resistant to at least one antibiotic, and twenty percent were resistant to at least two antibiotics. Three strains isolated from water samples (pond and inlet canal) exhibited multiresistance to ampicillin, tetracycline, oxytetracycline, and nitrofurantoin. One of them had a plasmid with genes conferring resistance to nitrofurantoin and ampicillin. The incidence of bacteria pathogenic to humans in a shrimp farming environment, as well as their drug-resistance pattern revealed in this study, emphasizes the need for a more rigorous attention to this area. Fátima C. T. Carvalho, Oscarina V. Sousa, Edirsana M. R. Carvalho, Ernesto Hofer, and Regine H. S. F. Vieira Copyright © 2013 Fátima C. T. Carvalho et al. All rights reserved. Effect of Commercial Cyanobacteria Products on the Growth and Antagonistic Ability of Some Bioagents under Laboratory Conditions Thu, 07 Nov 2013 18:01:31 +0000 Evaluation of the efficacy of blue-green algal compounds against the growth of either pathogenic or antagonistic microorganisms as well as their effect on the antagonistic ability of bioagents was studied under in vitro conditions. The present study was undertaken to explore the inhibitory effect of commercial algal compounds, Weed-Max and Oligo-Mix, against some soil-borne pathogens. In growth medium supplemented with these algal compounds, the linear growth of pathogenic fungi decreased by increasing tested concentrations of the two algal compounds. Complete reduction in pathogenic fungal growth was observed at 2% of both Weed-Max and Oligo-Mix. Gradual significant reduction in the pathogenic fungal growth was caused by the two bioagents and by increasing the concentrations of algal compounds Weed-Max and Oligo-Mix. The present work showed that commercial algal compounds, Weed-Max and Oligo-Mix, have potential for the suppression of soil-borne fungi and enhance the antagonistic ability of fungal, bacterial, and yeast bio-agents. Nehal S. El-Mougy and Mokhtar M. Abdel-Kader Copyright © 2013 Nehal S. El-Mougy and Mokhtar M. Abdel-Kader. All rights reserved. Proof of Principle for a Real-Time Pathogen Isolation Media Diagnostic: The Use of Laser-Induced Breakdown Spectroscopy to Discriminate Bacterial Pathogens and Antimicrobial-Resistant Staphylococcus aureus Strains Grown on Blood Agar Tue, 10 Sep 2013 08:18:28 +0000 Laser-Induced Breakdown Spectroscopy (LIBS) is a rapid, in situ, diagnostic technique in which light emissions from a laser plasma formed on the sample are used for analysis allowing automated analysis results to be available in seconds to minutes. This speed of analysis coupled with little or no sample preparation makes LIBS an attractive detection tool. In this study, it is demonstrated that LIBS can be utilized to discriminate both the bacterial species and strains of bacterial colonies grown on blood agar. A discrimination algorithm was created based on multivariate regression analysis of spectral data. The algorithm was deployed on a simulated LIBS instrument system to demonstrate discrimination capability using 6 species. Genetically altered Staphylococcus aureus strains grown on BA, including isogenic sets that differed only by the acquisition of mutations that increase fusidic acid or vancomycin resistance, were also discriminated. The algorithm successfully identified all thirteen cultures used in this study in a time period of 2 minutes. This work provides proof of principle for a LIBS instrumentation system that could be developed for the rapid discrimination of bacterial species and strains demonstrating relatively minor genomic alterations using data collected directly from pathogen isolation media. Rosalie A. Multari, David A. Cremers, Melissa L. Bostian, Joanne M. Dupre, and John E. Gustafson Copyright © 2013 Rosalie A. Multari et al. All rights reserved. Evaluation of Central Venous Catheter Associated Blood Stream Infections: A Microbiological Observational Study Tue, 09 Jul 2013 09:05:36 +0000 There are substantial morbidity and mortality associated with vascular catheter use among crictically ill patients. The attributable mortality is 10% to 25% which is associated with bacteremia among those who are hospitalized. This study was undertaken to identify catheter related blood stream infections, to isolate pathogenic microorganisms present in intravascular catheter related local infections, exit site infections, and to determine the predisposing factors for the development of such infections and antibiotic sensitivity pattern of the isolated organisms in tertiary care hospital. Vinay Khanna, Chiranjay Mukhopadhayay, Vandana K. E., Murlidhar Verma, and Partha Dabke Copyright © 2013 Vinay Khanna et al. All rights reserved. Genotypic Characterization of Yersinia enterocolitica Biotype 4/O:3 Isolates from Pigs and Slaughterhouses Using SE-AFLP, ERIC-PCR, and PFGE Thu, 30 May 2013 14:36:50 +0000 Yersinia enterocolitica is a foodborne pathogen that causes illness in humans and animals. The biotype 4/O:3 has been commonly associated with yersiniosis and is characterized by the presence of chromosomal and extra-chromosomal virulence genes. Molecular typing methods have been successfully used to characterize Y. enterocolitica genetic heterogeneity and to study the epidemiology of the bacteria from different origins. In this study, 320 Y. enterocolitica biotype 4/O:3 isolates originating in pigs and slaughterhouses were characterized according to the virulence profile, and 61 isolates were typified through SE-AFLP, ERIC-PCR, and PFGE techniques. The majority of the isolates originated from pigs, and the predominant virulence profile was ail+ virF+ rfbC+ ystA+, representing 83.4% of the tested isolates. All of the Y. enterocolitica 4/O:3 isolates were positive for at least ystA gene. The SE-AFLP and ERIC-PCR patterns were highly homogeneous. The SE-AFLP was more discriminative than the ERIC-PCR and tended to cluster isolates according to the slaughterhouse. Despite the limited genetic diversity of Y. enterocolitica 4/O:3, PFGE was shown to be the most discriminative technique considering one band of difference. Fattening pigs proved to be an important reservoir of Y. enterocolitica biotype 4/O:3 carrying virulence genes. Renata Paixão, Luisa Zanolli Moreno, Débora Dirani Sena de Gobbi, Daniele Cristine Raimundo, Thais Sebastiana Porfida Ferreira, Maria Garcia Spindola, Ernesto Hofer, Cristhiane Moura Falavina dos Reis, Maria Helena Matté, and Andrea Micke Moreno Copyright © 2013 Renata Paixão et al. All rights reserved.