International Journal of Microbiology The latest articles from Hindawi Publishing Corporation © 2016 , Hindawi Publishing Corporation . All rights reserved. Physicochemical and Antibacterial Properties of Chitosan Extracted from Waste Shrimp Shells Wed, 13 Jul 2016 06:00:23 +0000 This research aims to study the production of chitosan from shrimp shell (Litopenaeus vannamei) of waste origin using two chemical methodologies involving demineralization, deproteinization, and the degree of deacetylation. The evaluation of the quality of chitosan from waste shrimp shells includes parameters for the yield, physical chemistry characteristics by infrared spectroscopy (FT-IR), the degree of deacetylation, and antibacterial activity. The results showed (by Method 1) extraction yields for chitin of 33% and for chitosan of 49% and a 76% degree of deacetylation. Chitosan obtained by Method 2 was more efficient: chitin (36%) and chitosan (63%), with a high degree of deacetylation (81.7%). The antibacterial activity was tested against Gram-negative bacteria (Stenotrophomonas maltophilia and Enterobacter cloacae) and Gram-positive Bacillus subtilis and the Minimum Inhibitory Concentrations (MIC) and the Minimum Bactericidal Concentration (MBC) were determined. Method 2 showed that extracted chitosan has good antimicrobial potential against Gram-positive and Gram-negative bacteria and that the process is viable. José Carlos Vilar Junior, Daylin Rubio Ribeaux, Carlos Alberto Alves da Silva, and Galba Maria De Campos-Takaki Copyright © 2016 José Carlos Vilar Junior et al. All rights reserved. Pattern of Bacterial Pathogens and Their Susceptibility Isolated from Surgical Site Infections at Selected Referral Hospitals, Addis Ababa, Ethiopia Thu, 30 Jun 2016 13:55:29 +0000 Background. The emergence of multidrug resistant bacterial pathogens in hospitals is becoming a challenge for surgeons to treat hospital acquired infections. Objective. To determine bacterial pathogens and drug susceptibility isolated from surgical site infections at St. Paul Specialized Hospital Millennium Medical College and Yekatit 12 Referral Hospital Medical College, Addis Ababa, Ethiopia. Methods. A cross-sectional study was conducted between October 2013 and March 2014 on 107 surgical site infected patients. Wound specimens were collected using sterile cotton swab and processed as per standard operative procedures in appropriate culture media; and susceptibility testing was done using Kirby-Bauer disc diffusion technique. The data were analyzed by using SPSS version 20. Result. From a total of 107 swabs collected, 90 (84.1%) were culture positive and 104 organisms were isolated. E. coli (24 (23.1%)) was the most common organism isolated followed by multidrug resistant Acinetobacter species (23 (22.1%)). More than 58 (75%) of the Gram negative isolates showed multiple antibiotic resistance (resistance ≥ 5 drugs). Pan-antibiotic resistance was noted among 8 (34.8%) Acinetobacter species and 3 (12.5%) E. coli. This calls for abstinence from antibiotic abuse. Conclusion. Gram negative bacteria were the most important isolates accounting for 76 (73.1%). Ampicillin, amoxicillin, penicillin, cephazoline, and tetracycline showed resistance while gentamicin and ciprofloxacin were relatively effective antimicrobials. Walelign Dessie, Gebru Mulugeta, Surafael Fentaw, Amete Mihret, Mulu Hassen, and Engida Abebe Copyright © 2016 Walelign Dessie et al. All rights reserved. Group B Streptococcus Colonization among Pregnant Women Attending Antenatal Care at Tertiary Hospital in Rural Southwestern Uganda Sun, 22 May 2016 13:43:12 +0000 Objectives. This study sought to determine the prevalence and factors associated with group B streptococcal anogenital colonization among pregnant women attending antenatal care at Mbarara Regional Referral Hospital, a tertiary hospital. Methods. Cross-sectional study where 309 pregnant women ≥ thirty-five weeks of gestation attending antenatal clinic were consecutively recruited between January and March 2015. Anovaginal swabs were collected and tested qualitatively using rapid visual immunoassay GBS test kits for presence of GBS antigens. Data was analyzed using STATA version 12. In univariate analysis, GBS colonized mothers were presented as percentages and numbers, and in multivariate analysis logistic regression analysis was applied to determine the associations of exposure variable and GBS colonization; a value of less than 0.05 was considered significant. Results. Mothers’ median age was 25 years, 14.6% mothers being obese. GBS prevalence was 28.8%, 95% CI: 23.7–33.9. Obesity was the only significant factor associated with anogenital GBS colonization with odds ratio of 3.78, 95% CI: 1.78–8.35, a value of 0.001. Maternal ages, educational level, residence, and gravidity were not associated with GBS anogenital colonization. Conclusion. Group B streptococcal anogenital colonization among pregnant women attending antenatal care at tertiary hospital, in Southwestern Uganda, is high. Abdul Namugongo, Joel Bazira, Yarine Fajardot, and Ngonzi Joseph Copyright © 2016 Abdul Namugongo et al. All rights reserved. Antimicrobial Evaluation of Bacterial Isolates from Urine Specimen of Patients with Complaints of Urinary Tract Infections in Awka, Nigeria Wed, 20 Apr 2016 06:51:27 +0000 Urinary tract infections (UTIs) account for one of the major reasons for most hospital visits and the determination of the antimicrobial susceptibility patterns of uropathogens will help to guide physicians on the best choice of antibiotics to recommend to affected patients. This study is designed to isolate, characterize, and determine the antimicrobial susceptibility patterns of the pathogens associated with UTI in Anambra State Teaching Hospital, Amaku, Anambra State, Nigeria. Clean catch urine samples of inpatient and outpatient cases of UTI were collected and bacteriologically analyzed using standard microbiological procedures. Antibiogram was done by the Kirby-Bauer disc diffusion method. The most prevalent isolates were S. aureus (28%), E. coli (24.6%), and S. saprophyticus (20%). The antibacterial activities of the tested agents were in the order of Augmentin < Ceftazidime < Cefuroxime < Cefixime < Gentamicin < Ofloxacin < Ciprofloxacin < Nitrofurantoin. It was found that all the organisms were susceptible in varying degrees to Nitrofurantoin, Ciprofloxacin, and Ofloxacin. It was also observed that all the bacterial species except Streptococcus spp. have a Multiple Antibiotic Resistance Index (MARI) greater than 0.2. For empiric treatment of UTIs in Awka locality, Nitrofurantoin, Ciprofloxacin, and Ofloxacin are the first line of choice. Perpetua A. Ekwealor, Malachy C. Ugwu, Ifeanyi Ezeobi, George Amalukwe, Belinda C. Ugwu, Ugochukwu Okezie, Catherine Stanley, and Charles Esimone Copyright © 2016 Perpetua A. Ekwealor et al. All rights reserved. Novel Microdilution Method to Assess Double and Triple Antibiotic Combination Therapy In Vitro Mon, 18 Apr 2016 14:17:54 +0000 An in vitro microdilution method was developed to assess double and triple combinations of antibiotics. Five antibiotics including ciprofloxacin, amikacin, ceftazidime, piperacillin, and imipenem were tested against 10 clinical isolates of Pseudomonas aeruginosa. Each isolate was tested against ten double and nine triple combinations of the antibiotics. A 96-well plate was used to test three antibiotics, each one alone and in double and triple combinations against each isolate. The minimum bacteriostatic and bactericidal concentrations in combination were determined with respect to the most potent antibiotic. An Interaction Code (IC) was generated for each combination, where a numerical value was designated based on the 2-fold increase or decrease in the MICs with respect to the most potent antibiotic. The results of the combinations were verified by time-kill assay at constant concentrations of the antibiotics and in a chemostat. Only 13% of the double combinations were synergistic, whereas 5% showed antagonism. Forty-three percent of the triple combinations were synergistic with no antagonism observed, and 100% synergism was observed in combination of ciprofloxacin, amikacin, and ceftazidime. The presented protocol is simple and fast and can help the clinicians in the early selection of the effective antibiotic therapy for treatment of severe infections. Mohamed El-Azizi Copyright © 2016 Mohamed El-Azizi. All rights reserved. Detection of Extended-Spectrum Beta-Lactamase-Producing Escherichia coli in Market-Ready Chickens in Zambia Sun, 17 Apr 2016 12:09:01 +0000 The frequent administering of antibiotics in the treatment of poultry diseases may contribute to emergence of antimicrobial-resistant strains. The objective of this study was to detect the presence of extended-spectrum β-lactamase- (ESBL-) producing Escherichia coli in poultry in Zambia. A total of 384 poultry samples were collected and analyzed for ESBL-producing Escherichia coli. The cultured E. coli isolates were subjected to antimicrobial susceptibility tests and the polymerase chain reaction for detection of , , and genes. Overall 20.1%, 77/384, (95% CI; 43.2–65.5%) of total samples analyzed contained ESBL-producing Escherichia coli. The antimicrobial sensitivity test revealed that 85.7% (66/77; CI: 75.7–92) of ESBL-producing E. coli isolates conferred resistance to beta-lactam and other antimicrobial agents. These results indicate that poultry is a potential reservoir for ESBL-producing Escherichia coli. The presence of ESBL-producing Escherichia coli in poultry destined for human consumption requires strengthening of the antibiotic administering policy. This is important as antibiotic administration in food animals is gaining momentum for improved animal productivity in developing countries such as Zambia. K. Chishimba, B. M. Hang’ombe, K. Muzandu, S. E. Mshana, M. I. Matee, C. Nakajima, and Y. Suzuki Copyright © 2016 K. Chishimba et al. All rights reserved. Strains, Mechanism, and Perspective: Salmonella-Based Cancer Therapy Thu, 14 Apr 2016 12:25:22 +0000 Recently, investigation of bacterial-based tumor therapy has regained focus due to progress in molecular, cellular, and microbial biology. Many bacteria such as Salmonella, Listeria, Escherichia, and Clostridium have proved to have tumor targeting and in some cases even tumor-destroying phenotypes. Furthermore, bacterial clinical treatments for cancer have been improved by combination with other therapeutic methods such as chemotherapeutic drugs and radioactive agents. Synthetic biology techniques have also driven the development of new bacterial-based cancer therapies. However, basic questions about the mechanisms of bacterial-mediated tumor targeting and destruction are still being elucidated. In this review, we focus on three tumor-therapeutic Salmonella models, the most intensively studied bacterial genus in this field. One of these Salmonella models is our Salmonella enterica serovar Typhimurium LT2 derived strain CRC2631, engineered to minimize toxicity but maximize tumor-targeting and destruction effects. The other two are VNP20009 and A1-R. We compare the means by which these therapeutic candidate strain models were selected for study, their tumor targeting and tumor destruction phenotypes in vitro and in vivo, and what is currently known about the mechanisms by which they target and destroy tumors. Cheng-Zhi Wang, Robert A. Kazmierczak, and Abraham Eisenstark Copyright © 2016 Cheng-Zhi Wang et al. All rights reserved. Spread of TEM, VIM, SHV, and CTX-M β-Lactamases in Imipenem-Resistant Gram-Negative Bacilli Isolated from Egyptian Hospitals Thu, 31 Mar 2016 18:04:21 +0000 Carbapenem-resistant Gram-negative bacilli resulting from β-lactamases have been reported to be an important cause of nosocomial infections and are a critical therapeutic problem worldwide. This study aimed to describe the prevalence of imipenem-resistant Gram-negative bacilli isolates and detection of , , , , and genes in these clinical isolates in Egyptian hospitals. The isolates were collected from various clinical samples, identified by conventional methods and confirmed by API 20E. Antibiotic susceptibility testing was determined by Kirby-Bauer technique and interpreted according to CLSI. Production of , , , and genes was done by polymerase chain reaction (PCR). Direct sequencing from PCR products was subsequently carried out to identify and confirm these β-lactamases genes. Out of 65 isolates, (46.1%) Escherichia coli, (26.2%) Klebsiella pneumoniae, and (10.7%) Pseudomonas aeruginosa were identified as the commonest Gram-negative bacilli. 33(50.8%) were imipenem-resistant isolates. 22 isolates (66.7%) carried , 24(72.7%) had , and 5(15%) showed , while 12(36%), 6(18.2%), and 0(0.00%) harbored , , and , respectively. There is a high occurrence of β-lactamase genes in clinical isolates and sequence analysis of amplified genes showed differences between multiple SNPs (single nucleotide polymorphism) sites in the same gene among local isolates in relation to published sequences. El sayed Hamdy Mohammed, Ahmed Elsadek Fakhr, Hanan Mohammed El sayed, Said abd Elmohsen Al Johery, and Wesam Abdel Ghani Hassanein Copyright © 2016 El sayed Hamdy Mohammed et al. All rights reserved. IncF Plasmids Are Commonly Carried by Antibiotic Resistant Escherichia coli Isolated from Drinking Water Sources in Northern Tanzania Thu, 24 Mar 2016 15:37:03 +0000 The aim of this study was to identify the replicon types of plasmids, conjugation efficiencies, and the complement of antibiotic resistance genes for a panel of multidrug resistant E. coli isolates from surface waters in northern Tanzania. Standard membrane filtration was used to isolate and uidA PCR was used to confirm the identity of strains as E. coli. Antibiotic susceptibility was determined by breakpoint assay and plasmid conjugation was determined by filter-mating experiments. PCR and sequencing were used to identify resistance genes and PCR-based replicon typing was used to determine plasmid types. Filter mating experiments indicated conjugation efficiencies ranged from 10−1 to 10−7. Over 80% of the donor cells successfully passed their resistance traits and eleven different replicon types were detected (IncI1, FIC, P, FIIA, A/C, FIB, FIA, H12, K/B B/O, and N). IncF plasmids were most commonly detected (49% of isolates), followed by types IncI1 and IncA/C. Detection of these public health-relevant conjugative plasmids and antibiotic resistant traits in Tanzanian water suggests the possible pollution of these water sources from human, livestock, and wild animal wastes and also shows the potential of these water sources in the maintenance and transmission of these resistance traits between environments, animals, and people. Beatus Lyimo, Joram Buza, Murugan Subbiah, Sylivester Temba, Honest Kipasika, Woutrina Smith, and Douglas R. Call Copyright © 2016 Beatus Lyimo et al. All rights reserved. Diverse Molecular Genotypes of Mycobacterium tuberculosis Complex Isolates Circulating in the Free State, South Africa Thu, 17 Mar 2016 14:22:44 +0000 Tuberculosis is a serious public health concern especially in Africa and Asia. Studies describing strain diversity are lacking in the Free State region of South Africa. The aim of the study was to describe the diversity of Mycobacterium tuberculosis (M. tuberculosis) strain families in the Free State province of South Africa. A total of 86 M. tuberculosis isolates were genotyped using spoligotyping. A 12-locus mycobacterial interspersed repetitive units-variable-number tandem repeats (MIRU-VNTRs) typing was used to further characterize the resulting spoligotyping clusters. SITVITWEB identified 49 different patterns with allocation to six lineages including Latin-American-Mediterranean (LAM) (18 isolates), T (14 isolates), Beijing (five isolates), S (six isolates), Haarlem (one isolate), and X (five isolates), while 37 (43.0%) orphans were identified. Eight clusters included 37 isolates with identical spoligotypes (2 to 13/cluster). MIRU-VNTR typing further differentiated three spoligotyping clusters: SIT1/Beijing/MIT17, SIT33/LAM3/MIT213, and confirmed one SIT34/S/MIT311. In addition, SpolDB3/RIM assignment of the orphan strains resulted in a further 10 LAM and 13 T families. In total, LAM (28 isolates) and T (27 isolates) cause 63% of the individual cases of MTB in our study. The Free State has a highly diverse TB population with LAM being predominant. Further studies with inclusion of multidrug-resistant strains with larger sample size are warranted. Anneke Van der Spoel van Dijk, Pakiso M. Makhoahle, Leen Rigouts, and Kamaldeen Baba Copyright © 2016 Anneke Van der Spoel van Dijk et al. All rights reserved. Microbiological Quality Assessment by PCR and Its Antibiotic Susceptibility in Mangrove Crabs (Ucides cordatus) from Guanabara Bay, Rio de Janeiro, Brazil Tue, 15 Mar 2016 12:37:34 +0000 The bacteriological quality of crabs from three different mangroves (Itaóca, Suruí, and Piedade) from Rio de Janeiro state, Brazil, was investigated using conventional and molecular methods. The results revealed high counts for total coliforms in meat and hepatopancreas samples. PCR analyses identified 25 Escherichia coli colonies in the Itaóca, Piedade, and Suruí samples, detecting 13 enterotoxigenic colonies and 9 enteroaggregative colonies. Respectively, 12, 11, and 21 Vibrio parahaemolyticus strains were detected in the Itaóca, Piedade, and Suruí samples. Two V. cholerae strains were detected in the Piedade samples. The E. coli strains isolated in the present study showed resistance to gentamicin. E. coli strains from the Piedade samples showed 33% resistance to chloramphenicol and the strains also showed multiresistance to several antimicrobial agents with a MAR index ranging from 0.12 to 0.31. Vibrio strains from Piedade, Itaóca, and Suruí showed 86%, 78%, and 85% resistance, respectively, to ampicillin. The isolated Vibrio strains showed multiresistance to several antimicrobial agents, with a MAR index ranging from 0.12 to 0.25. The presence of these organisms in crab meat is an indication of microbial contamination, which may pose health risks to consumers when improperly cooked. M. C. N. Carvalho, M. M. Jayme, G. S. Arenazio, F. V. Araújo, S. G. F. Leite, and E. M. Del Aguila Copyright © 2016 M. C. N. Carvalho et al. All rights reserved. BK Virus Load Associated with Serum Levels of sCD30 in Renal Transplant Recipients Wed, 09 Mar 2016 11:20:37 +0000 Background. Rejection is the main drawback facing the renal transplant operations. Complicated and overlapping factors, mainly related to the immune system, are responsible for this rejection. Elevated serum levels of sCD30 were frequently recorded as an indicator for renal allograft rejection, while BV virus is considered as one of the most serious consequences for immunosuppressive treatment of renal transplant recipients (RTRs). Aims. This study aimed to determine the association of BK virus load with serum levels of sCD30 in RTRs suffering from nephropathy. Patients and Methods. A total of 50 RTRs with nephropathy and 30 age-matched apparently healthy individuals were recruited for this study. Serum samples were obtained from each participant. Real-time PCR was used to quantify BK virus load in RTRs serum, while ELISA technique was employed to estimate serum levels of sCD30. Results. Twenty-two percent of RTRs had detectable BKV with mean viral load of 1.094E + 06 ± 2.291E + 06. RTRs showed higher mean serum level of sCD30 ( U/mL) than that of controls ( U/mL) with significant difference. BK virus load had significant positive correlation with the serum levels of sCD30 in RTRs group. Conclusion. These results suggest that serum levels of sCD30 could be used as an indicator of BK viremia, and accordingly the immunosuppressive regime should be adjusted. Haidar A. Shamran, Salma N. Malik, Jinan M. Al-Saffer, and Rana S. Jawad Copyright © 2016 Haidar A. Shamran et al. All rights reserved. Bacteria in Cancer Therapy: Renaissance of an Old Concept Tue, 08 Mar 2016 15:37:28 +0000 The rising incidence of cancer cases worldwide generates an urgent need of novel treatment options. Applying bacteria may represent a valuable therapeutic variant that is intensively investigated nowadays. Interestingly, the idea to apply bacteria wittingly or unwittingly dates back to ancient times and was revived in the 19th century mainly by the pioneer William Coley. This review summarizes and compares the results of the past 150 years in bacteria mediated tumor therapy from preclinical to clinical studies. Lessons we have learned from the past provide a solid foundation on which to base future efforts. In this regard, several perspectives are discussed by which bacteria in addition to their intrinsic antitumor effect can be used as vector systems that shuttle therapeutic compounds into the tumor. Strategic solutions like these provide a sound and more apt exploitation of bacteria that may overcome limitations of conventional therapies. Sebastian Felgner, Dino Kocijancic, Michael Frahm, and Siegfried Weiss Copyright © 2016 Sebastian Felgner et al. All rights reserved. Synergistic Interaction of Methanol Extract from Canarium odontophyllum Miq. Leaf in Combination with Oxacillin against Methicillin-Resistant Staphylococcus aureus (MRSA) ATCC 33591 Tue, 23 Feb 2016 13:06:50 +0000 Canarium odontophyllum (CO) Miq. has been considered as one of the most sought-after plant species in Sarawak, Malaysia, due to its nutritional and pharmacological benefits. This study aimed to evaluate the pharmacodynamic interaction of crude methanol and acetone extracts from CO leaves in combination with oxacillin, vancomycin, and linezolid, respectively, against MRSA ATCC 33591 as preliminary study has reported its potential antistaphylococcal activity. The broth microdilution assay revealed that both methanol and acetone extracts were bactericidal with Minimum Inhibitory Concentration (MIC) of 312.5 μg/mL and 156.25 μg/mL and Minimum Bactericidal Concentration (MBC) of 625 μg/mL and 312.5 μg/mL, respectively. Fractional Inhibitory Concentration (FIC) indices were obtained via the chequerboard dilution assay where methanol extract-oxacillin, acetone extract-oxacillin, methanol extract-linezolid, and acetone extract-linezolid combinations exhibited synergism (FIC index ≤ 0.5). The synergistic action of the methanol extract-oxacillin combination was verified by time-kill analysis where bactericidal effect was observed at concentration of 1/8 × MIC of both compounds at 9.6 h compared to oxacillin alone. As such, these findings postulated that both extracts exert their anti-MRSA mechanism of action similar to that of vancomycin and provide evidence that the leaves of C. odontophyllum have the potential to be developed into antistaphylococcal agents. Dayang Fredalina Basri and Vimashiinee Sandra Copyright © 2016 Dayang Fredalina Basri and Vimashiinee Sandra. All rights reserved. Penicillin G-Induced Chlamydial Stress Response in a Porcine Strain of Chlamydia pecorum Sun, 21 Feb 2016 11:30:04 +0000 Chlamydia pecorum causes asymptomatic infection and pathology in ruminants, pigs, and koalas. We characterized the antichlamydial effect of the beta lactam penicillin G on Chlamydia pecorum strain 1710S (porcine abortion isolate). Penicillin-exposed and mock-exposed infected host cells showed equivalent inclusions numbers. Penicillin-exposed inclusions contained aberrant bacterial forms and exhibited reduced infectivity, while mock-exposed inclusions contained normal bacterial forms and exhibited robust infectivity. Infectious bacteria production increased upon discontinuation of penicillin exposure, compared to continued exposure. Chlamydia-induced cell death occurred in mock-exposed controls; cell survival was improved in penicillin-exposed infected groups. Similar results were obtained both in the presence and in the absence of the eukaryotic protein translation inhibitor cycloheximide and at different times of initiation of penicillin exposure. These data demonstrate that penicillin G induces the chlamydial stress response (persistence) and is not bactericidal, for this chlamydial species/strain in vitro, regardless of host cell de novo protein synthesis. Cory Ann Leonard, Frederic Dewez, and Nicole Borel Copyright © 2016 Cory Ann Leonard et al. All rights reserved. Distinct Habitats Select Particular Bacterial Communities in Mangrove Sediments Wed, 17 Feb 2016 10:47:50 +0000 We investigated the relationship among environmental variables, composition, and structure of bacterial communities in different habitats in a mangrove located nearby to an oil exploitation area, aiming to retrieve the natural pattern of bacterial communities in this ecosystem. The T-RFLP analysis showed a high diversity of bacterial populations and an increase in the bacterial richness from habitats closer to the sea and without vegetation (S1) to habitats covered by Avicennia schaueriana (S2) and Rhizophora mangle (S3). Environmental variables in S1 and S2 were more similar than in S3; however, when comparing the bacterial compositions, S2 and S3 shared more OTUs between them, suggesting that the presence of vegetation is an important factor in shaping these bacterial communities. In silico analyses of the fragments revealed a high diversity of the class Gammaproteobacteria in the 3 sites, although in general they presented quite different bacterial composition, which is probably shaped by the specificities of each habitat. This study shows that microhabitats inside of a mangrove ecosystem harbor diverse and distinct microbiota, reinforcing the need to conserve these ecosystems as a whole. Lidianne L. Rocha, Geórgia B. Colares, Vanessa L. R. Nogueira, Fernanda A. Paes, and Vânia M. M. Melo Copyright © 2016 Lidianne L. Rocha et al. All rights reserved. Hiding in Fresh Fruits and Vegetables: Opportunistic Pathogens May Cross Geographical Barriers Tue, 16 Feb 2016 16:19:38 +0000 Different microbial groups of the microbiome of fresh produce can have diverse effects on human health. This study was aimed at identifying some microbial communities of fresh produce by analyzing 105 samples of imported fresh fruits and vegetables originated from different countries in the world including local samples (Oman) for aerobic plate count and the counts of Enterobacteriaceae, Enterococcus, and Staphylococcus aureus. The isolated bacteria were identified by molecular (PCR) and biochemical methods (VITEK 2). Enterobacteriaceae occurred in 60% of fruits and 91% of vegetables. Enterococcus was isolated from 20% of fruits and 42% of vegetables. E. coli and S. aureus were isolated from 22% and 7% of vegetables, respectively. Ninety-seven bacteria comprising 21 species were similarly identified by VITEK 2 and PCR to species level. E. coli, Klebsiella pneumoniae, Enterococcus casseliflavus, and Enterobacter cloacae were the most abundant species; many are known as opportunistic pathogens which may raise concern to improve the microbial quality of fresh produce. Phylogenetic trees showed no relationship between clustering of the isolates based on the 16S rRNA gene and the original countries of fresh produce. Intercountry passage of opportunistic pathogens in fresh produce cannot be ruled out, which requires better management. Zahra S. Al-Kharousi, Nejib Guizani, Abdullah M. Al-Sadi, Ismail M. Al-Bulushi, and Baby Shaharoona Copyright © 2016 Zahra S. Al-Kharousi et al. All rights reserved. Activity of Medicinal Plant Extracts on Multiplication of Mycobacterium tuberculosis under Reduced Oxygen Conditions Using Intracellular and Axenic Assays Sun, 31 Jan 2016 11:21:02 +0000 Aim. Test the activity of selected medicinal plant extracts on multiplication of Mycobacterium tuberculosis under reduced oxygen concentration which represents nonreplicating conditions. Material and Methods. Acetone, ethanol and aqueous extracts of the plants Acorus calamus L. (rhizome), Ocimum sanctum L. (leaf), Piper nigrum L. (seed), and Pueraria tuberosa DC. (tuber) were tested on Mycobacterium tuberculosis H37Rv intracellularly using an epithelial cell (A549) infection model. The extracts found to be active intracellularly were further studied axenically under reducing oxygen concentrations. Results and Conclusions. Intracellular multiplication was inhibited ≥60% by five of the twelve extracts. Amongst these 5 extracts, in axenic culture, P. nigrum (acetone) was active under aerobic, microaerophilic, and anaerobic conditions indicating presence of multiple components acting at different levels and P. tuberosa (aqueous) showed bactericidal activity under microaerophilic and anaerobic conditions implying the influence of anaerobiosis on its efficacy. P. nigrum (aqueous) and A. calamus (aqueous and ethanol) extracts were not active under axenic conditions but only inhibited intracellular growth of Mycobacterium tuberculosis, suggesting activation of host defense mechanisms to mediate bacterial killing rather than direct bactericidal activity. Purva D. Bhatter, Pooja D. Gupta, and Tannaz J. Birdi Copyright © 2016 Purva D. Bhatter et al. All rights reserved. Comparison of Procleix Ultrio Elite and Procleix Ultrio NAT Assays for Screening of Transfusion Transmitted Infections among Blood Donors in India Tue, 19 Jan 2016 15:06:17 +0000 Background. Introduction of nucleic acid testing (NAT) has helped in decreasing window period donations, resulting in increased safety of blood supplies. NAT combines the advantages of direct and highly sequence-specific detection of viral genomes. We analysed the performance of newer Procleix Ultrio Elite (PUE) and Procleix Ultrio assay (PUA) for the screening of the viral markers in our donor population. Material and Methods. 10,015 donor samples were screened by routine immunoassays and both versions of NAT. NAT yields detected were subjected to viral load estimation and to other serological markers. Results. A total of 21 NAT yields were detected; three were positive by both NAT systems, whereas 18 samples were reactive by PUE only. NAT yields include 18 HBV and 3 HCV yields, of which 17 HBV yields were occult infections and 1 was window period (WP) infection. All 3 HCV yields were WP infections. No HIV-1/HIV-2 yield was found. Conclusion. Efficient target capture chemistry in the new TMA assay version significantly improved sensitivity. NAT is superior to serological immunoassays for screening of the viral markers; and the efficient target capture system in the newer TMA assay, namely, the PUE system, has significantly improved sensitivity over the earlier versions. Rahul Chaurasia, Diptiranjan Rout, Shamsuz Zaman, Kabita Chatterjee, Hem Chandra Pandey, and Abhishek Kumar Maurya Copyright © 2016 Rahul Chaurasia et al. All rights reserved. Antibiotic-Resistant Bacteria Are Major Threats of Otitis Media in Wollo Area, Northeastern Ethiopia: A Ten-Year Retrospective Analysis Tue, 19 Jan 2016 12:30:21 +0000 Antibiotic resistance is an increasingly serious threat to human health that needs an urgent action. The aim of this study was to determine the prevalence and antibiotic susceptibility profiles of bacteria isolated from patient ear discharges suspected of otitis media. A retrospective analysis was performed using culture and antibiotic susceptibility test results of 1225 patients who visited Dessie Regional Health Research Laboratory from 2001 to 2011. Results showed a strong association () between age and the risk of acquiring middle ear infection. The predominant bacterial isolates were Proteus spp. (28.8%), Staphylococcus aureus (23.7%), and Pseudomonas spp. (17.2%). Most of the isolated bacteria showed high resistance to ampicillin (88.5%), ceftriaxone (84.5%), amoxicillin (81.9%), and tetracycline (74.5%). About 72.5% of Proteus spp. and 62.2% of Pseudomonas spp. have developed resistance to one and more antibiotics used to treat them. This retrospective study also revealed the overall antibiotic resistance rate of bacterial isolates was increased nearly twofold () over the last decade. Relatively, ciprofloxacin and gentamicin were the most effective antibiotics against all the isolates. In conclusion, antibiotic-resistant bacteria are alarmingly increasing in Wollo area, northeastern Ethiopia, and becoming a major public health problem in the management of patients with middle ear infection. Ayele Argaw-Denboba, Asrat Agalu Abejew, and Alemayehu Gashaw Mekonnen Copyright © 2016 Ayele Argaw-Denboba et al. All rights reserved. Eucalyptus Tree: A Potential Source of Cryptococcus neoformans in Egyptian Environment Wed, 13 Jan 2016 11:28:48 +0000 In Egypt, the River Red Gum (Eucalyptus camaldulensis) is a well-known tree and is highly appreciated by the rural and urban dwellers. The role of Eucalyptus trees in the ecology of Cryptococcus neoformans is documented worldwide. The aim of this survey was to show the prevalence of C. neoformans during the flowering season of E. camaldulensis at the Delta region in Egypt. Three hundred and eleven samples out of two hundred Eucalyptus trees, including leaves, flowers, and woody trunks, were collected from four governorates in the Delta region. Thirteen isolates of C. neoformans were recovered from Eucalyptus tree samples (4.2%). Molecular identification of C. neoformans was done by capsular gene specific primer CAP64 and serotype identification was done depending on LAC1 gene. This study represents an update on the ecology of C. neoformans associated with Eucalyptus tree in Egyptian environment. Mahmoud Elhariri, Dalia Hamza, Rehab Elhelw, and Mohamed Refai Copyright © 2016 Mahmoud Elhariri et al. All rights reserved. Modulation of mecA Gene Expression by Essential Oil from Salvia sclarea and Synergism with Oxacillin in Methicillin Resistant Staphylococcus epidermidis Carrying Different Types of Staphylococcal Chromosomal Cassette mec Wed, 06 Jan 2016 14:01:08 +0000 The essential oil (EO) from Salvia sclarea was shown to increase the susceptibility of methicillin resistant Staphylococcus epidermidis (MRSE) isolates to oxacillin. The purpose of this study was to investigate the effect of EO from S. sclarea on expression of mecA gene of MRSE carrying different types of staphylococcal chromosomal cassette (SCCmec) and to evaluate potential synergistic effect of EO with oxacillin. Using real-time PCR we found that EO alone inhibited the expression of the resistant genes mecA, mecR1, and mecI and blaZ, blaR1, and blaI. The use of the combination of EO with oxacillin resulted in significantly inhibited expression of mecA gene in all tested strains with different types of SCCmec. Using time-kill assay and checkerboard assay we confirmed synergistic effect of EO from S. sclarea and oxacillin in MRSE. Romana Chovanová, Mária Mikulášová, and Štefánia Vaverková Copyright © 2016 Romana Chovanová et al. All rights reserved. Spectrum and the In Vitro Antifungal Susceptibility Pattern of Yeast Isolates in Ethiopian HIV Patients with Oropharyngeal Candidiasis Tue, 05 Jan 2016 13:42:29 +0000 Background. In Ethiopia, little is known regarding the distribution and the in vitro antifungal susceptibility profile of yeasts. Objective. This study was undertaken to determine the spectrum and the in vitro antifungal susceptibility pattern of yeasts isolated from HIV infected patients with OPC. Method. Oral pharyngeal swabs taken from oral lesions of study subjects were inoculated onto Sabouraud Dextrose Agar. Yeasts were identified by employing conventional test procedures and the susceptibility of yeasts to antifungal agents was evaluated by disk diffusion assay method. Result. One hundred and fifty-five yeast isolates were recovered of which 91 isolates were from patients that were not under HAART and 64 were from patients that were under HAART. C. albicans was the most frequently isolated species followed by C. glabrata, C. tropicalis, C. krusei, C. kefyr, Cryptococcus laurentii, and Rhodotorula species. Irrespective of yeasts isolated and identified, 5.8%, 5.8%, 12.3%, 8.4%, 0.6%, and 1.3% of the isolates were resistant to amphotericin B, clotrimazole, fluconazole, ketoconazole, miconazole, and nystatin, respectively. Conclusion. Yeast colonization rate of 69.2% and 31% resistance to six antifungal agents was documented. These highlight the need for nationwide study on the epidemiology of OPC and resistance to antifungal drugs. Birhan Moges, Adane Bitew, and Aster Shewaamare Copyright © 2016 Birhan Moges et al. All rights reserved. In Vitro Antimicrobial Activity of Spices and Medicinal Herbs against Selected Microbes Associated with Juices Mon, 04 Jan 2016 12:49:29 +0000 In the present investigation, comparison of antimicrobial activities of different spices, Curcuma longa, Zingiber officinale, and Mentha arvensis, and medicinal herbs, such as Withania somnifera, Rauvolfia serpentina, Emblica officinalis, Terminalia arjuna, and Centella asiatica, was evaluated. Different extraction solvents (acetone, methanol, ethanol, and water) were used and extracts were examined against Bacillus cereus, Serratia sp., Rhodotorula mucilaginosa, Aspergillus flavus, and Penicillium citrinum isolated from juices. Extracts from the medicinal herb and spices have significant activity. B. cereus was the most sensitive and R. mucilaginosa was the most resistant among the microorganisms tested. Ethanolic and methanolic extract of C. asiatica displayed maximum diameter of inhibition zone against bacteria and yeast and percentage mycelial inhibition against moulds. This study confirmed the potential of selected extracts of spices as effective natural food preservative in juices. Romika Dhiman, Neeraj Aggarwal, Kamal Rai Aneja, and Manpreet Kaur Copyright © 2016 Romika Dhiman et al. All rights reserved. In Vitro Antimicrobial Activity of Essential Oil of Thymus schimperi, Matricaria chamomilla, Eucalyptus globulus, and Rosmarinus officinalis Mon, 04 Jan 2016 11:10:38 +0000 In this study, the in vitro antimicrobial activities of four plant essential oils (T. schimperi, E. globulus, R. officinalis, and M. Chamomilla) were evaluated against bacteria and fungi. The studies were carried out using agar diffusion method for screening the most effective essential oils and agar dilution to determine minimum inhibitory concentration of the essential oils. Results of this study revealed that essential oils of T. schimperi, E. globulus, and R. officinalis were active against bacteria and some fungi. The antimicrobial effect of M. chamomilla was found to be weaker and did not show any antimicrobial activity. The minimum inhibitory concentration values of T. schimperi were 15.75 mg/mL for most of the bacteria and fungi used in this study. The minimum inhibitory concentration values of the other essential oils were in the range of 15.75–36.33 mg/mL against tested bacteria. This study highlighted the antimicrobial activity of the essential oil of E. globulus, M. chamomilla, T. Schimperi, and R. officinalis. The results indicated that T. schimperi have shown strong antimicrobial activity which could be potential candidates for preparation of antimicrobial drug preparation. Awol Mekonnen, Berhanu Yitayew, Alemnesh Tesema, and Solomon Taddese Copyright © 2016 Awol Mekonnen et al. All rights reserved. Rapid Accumulation of Total Lipid in Rhizoclonium africanum Kutzing as Biodiesel Feedstock under Nutrient Limitations and the Associated Changes at Cellular Level Thu, 31 Dec 2015 09:35:29 +0000 Increase of total lipid and the proportion of the favorable fatty acids in marine green filamentous macroalga Rhizoclonium africanum (Chlorophyceae) was studied under nitrate and phosphate limitations. These stresses were given by both eliminating and doubling the required amounts of nitrate and phosphate salts in the growth media. A significant twofold increase in total lipid (193.03 mg/g) was achieved in cells in absence of nitrate in the culture medium, followed by phosphate limitation (142.65 mg/g). The intracellular accumulation of neutral lipids was observed by fluorescence microscopy. The scanning electron microscopic study showed the major structural changes under nutrient starvation. Fourier transform infrared spectroscopy (FTIR) revealed the presence of ester (C-O-C stretching), ketone (C-C stretching), carboxylic acid (O-H bending), phosphine (P-H stretching), aromatic (C-H stretching and bending), and alcohol (O-H stretching and bending) groups in the treated cells indicating the high accumulation of lipid hydrocarbons in the treated cells. Elevated levels of fatty acids favorable for biodiesel production, that is, C16:0, C16:1, C18:1, and C20:1, were identified under nitrate- and phosphate-deficient conditions. This study shows that the manipulation of cultural conditions could affect the biosynthetic pathways leading to increased lipid production while increasing the proportion of fatty acids suitable for biodiesel production. Gour Gopal Satpati, Sanjit Kanjilal, Rachapudi Badari Narayana Prasad, and Ruma Pal Copyright © 2015 Gour Gopal Satpati et al. All rights reserved. Carbon Sources for Yeast Growth as a Precondition of Hydrogen Peroxide Induced Hormetic Phenotype Wed, 30 Dec 2015 09:18:26 +0000 Hormesis is a phenomenon of particular interest in biology, medicine, pharmacology, and toxicology. In this study, we investigated the relationship between H2O2-induced hormetic response in S. cerevisiae and carbon sources in yeast growth medium. In general, our data indicate that (i) hydrogen peroxide induces hormesis in a concentration-dependent manner; (ii) the effect of hydrogen peroxide on yeast reproductive ability depends on the type of carbon substrate in growth medium; and (iii) metabolic and growth rates as well as catalase activity play an important role in H2O2-induced hormetic response in yeast. Ruslana Vasylkovska, Natalia Petriv, and Halyna Semchyshyn Copyright © 2015 Ruslana Vasylkovska et al. All rights reserved. Isolation of Bacillus sphaericus from Lombok Island, Indonesia, and Their Toxicity against Anopheles aconitus Mon, 14 Dec 2015 09:15:47 +0000 Malaria is endemic to Lombok Island, Indonesia. One approach to suppress malaria spread is to eliminate anopheline larvae in their habitat and the environmentally safe agent is bacteria, that is, Bacillus sphaericus. However, there is no information regarding local isolate of B. sphaericus that is toxic to mosquito larvae from Lombok. The aims of the study were to isolate B. sphaericus from soil in areas close to beach surrounding Lombok Island and to test their toxicity against 3rd instar Anopheles aconitus larvae. Soil samples were collected from 20 different sampling locations from Lombok Island and homogenized with sterile physiological salt solution. Suspension was heat-shocked at 80°C for 30 minutes and then spread onto antibiotic-supplemented NYSM solid medium. Colonies grown were characterized and subjected to initial toxicity test against anopheline larvae. Isolates with more than 50% killing percentage were subjected to bioassay testing against anopheline larvae. From 20 locations, 1 isolate showed mild toxicity (namely, isolate MNT) and 2 isolates showed high toxicity (namely, isolates SLG and TJL2) against An. aconitus. Those 3 isolates were potentially useful isolates, as they killed almost all larvae in 24 hours. The discovery of toxic indigenous isolates of B. sphaericus from Lombok Island opens opportunity to develop a biopesticide from local resources. Bambang Fajar Suryadi, Bagyo Yanuwiadi, Tri Ardyati, and Suharjono Copyright © 2015 Bambang Fajar Suryadi et al. All rights reserved. In Vitro Evaluation of Antimicrobial Efficacy of Extracts Obtained from Raw and Fermented Wild Macrofungus, Lenzites quercina Thu, 29 Oct 2015 11:00:38 +0000 In recent time, there is a major concern about antibiotic resistance displayed by some pathogenic microorganisms and this had involved a continuous search for natural antimicrobial products. The phytochemistry as well as antimicrobial activity of extracts obtained from Lenzites quercina was investigated. The extracts and purified fractions were, respectively, tested against indicator organisms using agar well diffusion and disc diffusion methods. The quantity of phytochemicals found in the extracts of L. quercina ranged from 14.4 to 20.7 mg/g for alkaloids, 6.1 to 12.8 mg/g for steroids, 4.5 to 10.6 mg/g for saponins, 2.8 to 17.2 mg/g for terpenoids, and 0.41 to 17.1 mg/g for flavonoids. The gas chromatography mass spectrophotometry (GCMS) analysis of the extract reveals the presence of caprylic acid, stearic acid, tetradecanoic acid, methyl-11-octadecenoate, oleic acid, and 4-methyl-2-propyl-1-pentanol. Extracts of L. quercina and its purified fractions exhibited wider range of inhibition (4 mm to 26 mm) on Staphylococcus aureus (ATCC 29213), Pseudomonas aeruginosa (ATCC 27853), Escherichia coli (ATCC 35218), Methicillin Resistant Staphylococcus aureus (MRSA), Salmonella typhi, Bacillus cereus, Enterococcus faecalis, Candida albicans, and Aspergillus niger. The antimicrobial effects of L. quercina extracts indicate that this wild macrofungus contains significant amount of pharmacological agents, which could be extracted to curb the menace of antibiotic resistances by pathogenic organisms. Olusola Clement Ogidi, Victor Olusegun Oyetayo, and Bamidele Juliet Akinyele Copyright © 2015 Olusola Clement Ogidi et al. All rights reserved. Nosocomial Isolates and Their Drug Resistant Pattern in ICU Patients at National Institute of Neurological and Allied Sciences, Nepal Mon, 26 Oct 2015 06:17:58 +0000 Multidrug resistant organisms are increasing day by day and the cause is poorly known. This study was carried out from June 2011 to May 2012 at National Institute of Neurological and Allied Sciences Kathmandu, Nepal, with a view to determining drug resistant pathogens along with detection of extended spectrum β-lactamase (ESBL), AmpC β-lactamase (ABL), and metallo-β-lactamase (MBL) producing bacteria causing infection to ICU patients. A standard methodology was used to achieve these objectives as per recommendation of American Society for Microbiology. ESBL was detected by combined disc assay using cefotaxime and cefotaxime clavulanic acid, ABL by inhibitor based method using cefoxitin and phenylboronic acid, and MBL by imipenem-EDTA combined disk method. Two hundred and ninety-four different clinical samples such as tracheal aspirates, urine, pus, swabs, catheter tips, and blood were processed during the study. Most common bacteria were Acinetobacter spp. Of the total 58 Acinetobacter spp., 46 (79%) were MDR, and 27% were positive for ABL and 12% were for MBL. Of the 32 cases of Staphylococcus aureus, 18 (56%) were MDR. Findings of this study warrant routine β-lactamase testing in clinical isolates. Pashupati Bhandari, Ganesh Thapa, Bharat Mani Pokhrel, Dwij Raj Bhatta, and Upendra Devkota Copyright © 2015 Pashupati Bhandari et al. All rights reserved.