International Journal of Microbiology The latest articles from Hindawi Publishing Corporation © 2016 , Hindawi Publishing Corporation . All rights reserved. Audit of Helicobacter pylori Testing in Microbiology Laboratories in England: To Inform Compliance with NICE Guidance and the Feasibility of Routine Antimicrobial Resistance Surveillance Tue, 18 Oct 2016 07:13:31 +0000 Introduction. The National Institute for Health and Clinical Excellence (NICE) guidance recommends that dyspeptic patients are tested for Helicobacter pylori using a urea breath test, stool antigen test, or serology. Antibiotic resistance in H. pylori is globally increasing, but treatment in England is rarely guided by susceptibility testing or surveillance. Aims. To determine compliance of microbiology laboratories in England with NICE guidance and whether laboratories perform culture and antibiotic susceptibility testing (AST). Methods. In 2015, 170 accredited English microbiology laboratories were surveyed, by email. Results. 121/170 (71%) laboratories responded; 96% provided H. pylori testing (78% on site). 94% provided H. pylori diagnosis using stool antigen; only four provided serology as their noninvasive test; 3/4 of these encouraged urea breath tests in their acute trusts. Only 22/94 (23%) of the laboratories performed H. pylori cultures from gastric biopsies on site; 9/22 performed AST, but the vast majority processed less than one specimen/week. Conclusions. Only five laboratories in England do not comply with NICE guidance; these will need the guidance reinforced. National surveillance needs to be implemented; culture-based AST would need to be centralised. Moving forward, detection of resistance in H. pylori from stool specimens using molecular methods (PCR) needs to be explored. Rosalie Allison, Donna M. Lecky, Megan Bull, Kim Turner, Gauri Godbole, and Cliodna A. M. McNulty Copyright © 2016 Rosalie Allison et al. All rights reserved. Biological Characteristics and Antimicrobial Activity of Endophytic Streptomyces sp. TQR12-4 Isolated from Elite Citrus nobilis Cultivar Ham Yen of Vietnam Tue, 04 Oct 2016 11:21:27 +0000 Ham Yen orange (Citrus nobilis Lour) is the highly valuable commercial fruit of Vietnam. With the blooming of fruit production and farming area, this specialty crop is facing threats from several serious diseases; therefore the search for new effective biocontrollers is required to prevent the existing excessive use of fertilizers and plant protection chemicals. Endophytic actinomycetes are of great scientific interest due to their high potential of application in agriculture and pharmaceutical research. In this work, endophytic actinomycetes were isolated from a native orange species of Northeast mountainous province Tuyen Quang. Among 49 isolates obtained, the isolate TQR12-4 strongly inhibited test pathogens Colletotrichum truncatum, Geotrichum candidum, Fusarium oxysporum, and F. udum. This isolate gave comparatively high biomass yields on different substrates, for example, carboxy methyl cellulose, starch, protein, and chitin, within a wide range of temperature from 15 to 45°C and pH from 4 to 10. Sequence analysis of 16S rDNA gene showed that TQR12-4 shared 99% similarity to Streptomyces prasinopilosus; however, it slightly differed from the latter in spore morphology and hence was named as Streptomyces sp. TQR12-4. A thermostable antifungal substance of nonpeptide nature produced by Streptomyces sp. TQR12-4 had MIC against Fusarium udum of 100 μg/mL and 400 μg/mL respective to extract fractions and . Phan Thi Hong-Thao, Nguyen Vu Mai-Linh, Nguyen Thi Hong-Lien, and Nguyen Van Hieu Copyright © 2016 Phan Thi Hong-Thao et al. All rights reserved. Antibiotic Resistance of Diverse Bacteria from Aquaculture in Borneo Sun, 25 Sep 2016 11:15:54 +0000 The administration of antimicrobials in aquaculture provides a selective pressure creating a reservoir of multiple resistant bacteria in the cultured fish and shrimps as well as the aquaculture environment. The objective of this study was to determine the extent of antibiotic resistance in aquaculture products and aquaculture’s surrounding environment in Sarawak, Malaysian Borneo. Ninety-four identified bacterial isolates constituted of 17 genera were isolated from sediment, water, and cultured organisms (fish and shrimp) in selected aquaculture farms. These isolates were tested for their antibiotic resistance against 22 antibiotics from several groups using the disk diffusion method. The results show that the highest resistance was observed towards streptomycin (85%, ), while the lowest resistance was towards gentamicin (1.1%, ). The multiple antibiotic resistant (MAR) index of the isolates tested ranged between 0 and 0.63. It was suggested that isolates with MAR index > 0.2 were recovered from sources with high risk of antibiotic resistant contamination. This study revealed low level of antibiotic resistance in the aquaculture bacterial isolates except for streptomycin and ampicillin (>50% resistance, ) which have been used in the aquaculture industry for several decades. Antibiotic resistant patterns should be continuously monitored to predict the emergence and widespread of MAR. Effective action is needed to keep the new resistance from further developing and spreading. M. M. Kathleen, L. Samuel, C. Felecia, E. L. Reagan, A. Kasing, M. Lesley, and S. C. Toh Copyright © 2016 M. M. Kathleen et al. All rights reserved. Impact of Some Ecological Factors on Fecal Contamination of Drinking Water by Diarrheagenic Antibiotic-Resistant Escherichia coli in Zagazig City, Egypt Tue, 20 Sep 2016 14:26:33 +0000 Fecal contamination of drinking water is a major health problem which accounts for many cases of diarrhea mainly in infants and foreigners. This contamination is a complex interaction of many parameters. Antibiotic resistance among bacterial isolates complicates the problem. The study was done to identify fecal contamination of drinking water by Diarrheagenic Antibiotic-Resistant Escherichia coli in Zagazig city and to trace reasons for such contamination, three hundred potable water samples were investigated for E. coli existence. Locations of E. coli positive samples were investigated in relation to population density, water source, and type of water pipe. Sixteen E. coli strains were isolated. Antibiotic sensitivity was done and enterotoxigenic, enteropathogenic, and enterohaemorrhagic virulence genes were investigated by PCR. Probability of fecal contamination correlated with higher population density, with increased distance from Zagazig water plant, and with asbestos cement water pipes. Resistance to at least one antimicrobial drug was found in all isolates. Virulence genes were detected in a rate of 26.27%, 13.13%, 20%, 6.67%, and 33.33% for LT, ST, stx1, stx2, and eae genes, respectively. This relatively high frequency of fecal contamination points towards the high risk of developing diarrhea by antibiotic resistant DEC in low socioeconomic communities particularly with old fashion distribution systems. Ahmed Elsadek Fakhr, Maha Kamal Gohar, and Amal Hassan Atta Copyright © 2016 Ahmed Elsadek Fakhr et al. All rights reserved. Chemical Composition and Antibacterial Activity of Essential Oils of Tagetes minuta (Asteraceae) against Selected Plant Pathogenic Bacteria Sun, 18 Sep 2016 09:46:19 +0000 The objective of this study was to determine the chemical composition and antibacterial activity of essential oils (EOs) of Tagetes minuta against three phytopathogenic bacteria Pseudomonas savastanoi pv. phaseolicola, Xanthomonas axonopodis pv. phaseoli, and Xanthomonas axonopodis pv. manihotis. The essential oils were extracted using steam distillation method in a modified Clevenger-type apparatus while antibacterial activity of the EOs was evaluated by disc diffusion method. Gas chromatography coupled to mass spectrometry (GC/MS) was used for analysis of the chemical profile of the EOs. Twenty compounds corresponding to 96% of the total essential oils were identified with 70% and 30% of the identified components being monoterpenes and sesquiterpenes, respectively. The essential oils of T. minuta revealed promising antibacterial activities against the test pathogens with Pseudomonas savastanoi pv. phaseolicola being the most susceptible with mean inhibition zone diameters of 41.83 and 44.83 mm after 24 and 48 hours, respectively. The minimum inhibitory concentrations and minimum bactericidal concentrations of the EOs on the test bacteria were in the ranges of 24–48 mg/mL and 95–190 mg/mL, respectively. These findings provide a scientific basis for the use of T. minuta essential oils as a botanical pesticide for management of phytopathogenic bacteria. Martin Muthee Gakuubi, John M. Wagacha, Saifuddin F. Dossaji, and Wycliffe Wanzala Copyright © 2016 Martin Muthee Gakuubi et al. All rights reserved. Comment on “Bacteria in Cancer Therapy: Renaissance of an Old Concept” Wed, 14 Sep 2016 09:23:29 +0000 Amin Talebi Bezmin Abadi Copyright © 2016 Amin Talebi Bezmin Abadi. All rights reserved. Bacterial Etiology and Antibiotic Resistance Profile of Community-Acquired Urinary Tract Infections in a Cameroonian City Wed, 07 Sep 2016 16:45:28 +0000 Introduction. Community-acquired urinary tract infections (CAUTIs) are usually treated empirically. Geographical variations in etiologic agents and their antibiotic sensitivity patterns are common. Knowledge of antibiotic resistance trends is important for improving evidence-based recommendations for empirical treatment of UTIs. Our aim was to determine the major bacterial etiologies of CAUTIs and their antibiotic resistance patterns in a cosmopolitan area of Cameroon for comparison with prescription practices of local physicians. Methods. We performed a cross-sectional descriptive study at two main hospitals in Yaoundé, collecting a clean-catch mid-stream urine sample from 92 patients having a clinical diagnosis of UTI. The empirical antibiotherapy was noted, and identification of bacterial species was done on CLED agar; antibiotic susceptibility testing was performed using the Kirby-Bauer disc diffusion method. Results. A total of 55 patients had samples positive for a UTI. Ciprofloxacin and amoxicillin/clavulanic acid were the most empirically prescribed antibiotics (30.9% and 23.6%, resp.); bacterial isolates showed high prevalence of resistance to both compounds. Escherichia coli (50.9%) was the most common pathogen, followed by Klebsiella pneumoniae (16.4%). Prevalence of resistance for ciprofloxacin was higher compared to newer quinolones. Conclusions. E. coli and K. pneumoniae were the predominant bacterial etiologies; the prevalence of resistance to commonly prescribed antibiotics was high. Rolf Nyah-tuku Nzalie, Hortense Kamga Gonsu, and Sinata Koulla-Shiro Copyright © 2016 Rolf Nyah-tuku Nzalie et al. All rights reserved. Phosphate Solubilization Potential of Rhizosphere Fungi Isolated from Plants in Jimma Zone, Southwest Ethiopia Mon, 05 Sep 2016 11:43:45 +0000 Phosphorus (P) is one of the major bioelements limiting agricultural production. Phosphate solubilizing fungi play a noteworthy role in increasing the bioavailability of soil phosphates for plants. The present study was aimed at isolating and characterizing phosphate solubilizing fungi from different rhizospheres using both solid and liquid Pikovskaya (PVK) medium. A total of 359 fungal isolates were obtained from 150 rhizosphere soil samples of haricot bean, faba bean, cabbage, tomato, and sugarcane. Among the isolates, 167 (46.52%) solubilized inorganic phosphate. The isolated phosphate solubilizing fungi belonged to genera of Aspergillus (55.69%), Penicillium spp. (23.35%), and Fusarium (9.58%). Solubilization index (SI) ranged from 1.10 to 3.05. Isolates designated as JUHbF95 (Aspergillus sp.) and JUFbF59 (Penicillium sp.) solubilized maximum amount of P 728.77 μg·mL−1 and 514.44 μg mL−1, respectively, from TCP (tricalcium phosphate) after 15 days of incubation. The highest (363 μg mL−1) soluble-P was released from RP with the inoculation of JUHbF95 in the PVK broth after 10 days of incubation. The present study indicated the presence of diverse plant associated P-solubilizing fungi that may serve as potential biofertilizers. Firew Elias, Delelegn Woyessa, and Diriba Muleta Copyright © 2016 Firew Elias et al. All rights reserved. Antibacterial and Antibiofilm Effect of Low Viscosity Chitosan against Staphylococcus epidermidis Thu, 18 Aug 2016 16:34:25 +0000 Aim. The aim of this study was to investigate the antibacterial and antibiofilm properties of low viscosity chitosan on S. epidermidis growth and biofilm formation. Methods and Results. The antibacterial and antibiofilm properties were investigated, during both planktonic growth and biofilm formation. This was performed using different concentrations in media and by coating on polystyrene surfaces. In addition, the bactericidal effect was investigated using a modified direct contact test. The results showed that low viscosity chitosan in media had both a bacteriostatic and bactericidal effect on planktonic growth and biofilm formation of S. epidermidis in a concentration dependent manner. Polystyrene discs coated with chitosan reduced both early biofilm formation (6 h) and late biofilm formation (18 h), as confirmed by scanning electron microscopy. The modified direct contact test showed a bactericidal effect. Conclusion. This study demonstrated that low viscosity chitosan has a bacteriostatic and bactericidal activity against S. epidermidis and that the activity is dependent on the amount of chitosan added. In addition, low viscosity chitosan reduced biofilm formation both when added to media and when coated on polystyrene surfaces. Significance and Impact of Study. Low viscosity chitosan could be a contribution to new treatment approaches of biofilm-related infections of S. epidermidis. Inger Sofie Dragland, Håkon Valen Rukke, Ida S. R. Stenhagen, Jessica Lönn-Stensrud, and Hilde M. Kopperud Copyright © 2016 Inger Sofie Dragland et al. All rights reserved. Identification of Common Bacterial Pathogens Causing Meningitis in Culture-Negative Cerebrospinal Fluid Samples Using Real-Time Polymerase Chain Reaction Mon, 01 Aug 2016 13:51:27 +0000 Background. Meningitis is a serious communicable disease with high morbidity and mortality rates. It is an endemic disease in Egypt caused mainly by Streptococcus pneumoniae, Neisseria meningitidis, and Haemophilus influenzae. In some settings, bacterial meningitis is documented depending mainly on positive cerebrospinal fluid (CSF) culture results or CSF positive latex agglutination test, missing the important role of prior antimicrobial intake which can yield negative culture and latex agglutination test results. This study aimed to utilize molecular technology in order to diagnose bacterial meningitis in culture-negative CSF samples. Materials and Methods. Forty culture-negative CSF samples from suspected cases of bacterial meningitis were examined by real-time polymerase chain reaction (real-time PCR) for the presence of lytA, bexA, and ctrA genes specific for Streptococcus pneumoniae, Haemophilus influenzae, and Neisseria meningitidis, respectively. Results. Positive real-time PCR results for Streptococcus pneumoniae were detected in 36 (90%) of culture-negative CSF samples while no positive results for Haemophilus influenzae or Neisseria meningitidis were detected. Four (10%) samples were negative by real-time PCR for all tested organisms. Conclusion. The use of molecular techniques as real-time PCR can provide a valuable addition to the proportion of diagnosed cases of bacterial meningitis especially in settings with high rates of culture-negative results. Walaa Shawky Khater and Safia Hamed Elabd Copyright © 2016 Walaa Shawky Khater and Safia Hamed Elabd. All rights reserved. Microbial Content of “Bowl Water” Used for Communal Handwashing in Preschools within Accra Metropolis, Ghana Sun, 31 Jul 2016 13:46:20 +0000 Objective. This study aimed at determining the microbial content of “bowl water” used for communal handwashing in preschools within the Accra Metropolis. Method. Six (6) preschools in the Accra Metropolis were involved in the study. Water samples and swabs from the hands of the preschool children were collected. The samples were analysed and tested for bacteria, fungi, parasites, and rotavirus. Results. Eight different bacteria, two different parasites, and a fungus were isolated while no rotavirus was detected. Unlike the rest of the microbes, bacterial isolates were found among samples from all the schools, with Staphylococcus species being the most prevalent (40.9%). Out of the three schools that had parasites in their water, two of them had Cryptosporidium parvum. The fungus isolated from two out of the six schools was Aspergillus niger. All bacteria isolated were found to be resistant to cotrimoxazole, ciprofloxacin, and ampicillin and susceptible to amikacin and levofloxacin. Conclusion. Although handwashing has the ability to get rid of microbes, communal handwashing practices using water in bowls could be considered a possible transmission route and may be of public concern. Patience B. Tetteh-Quarcoo, Isaac Anim-Baidoo, Simon Kwaku Attah, Bawa Abdul-Latif Baako, Japheth A. Opintan, Andrew A. Minamor, Mubarak Abdul-Rahman, and Patrick F. Ayeh-Kumi Copyright © 2016 Patience B. Tetteh-Quarcoo et al. All rights reserved. Influence of Vinasse Application in the Structure and Composition of the Bacterial Community of the Soil under Sugarcane Cultivation Tue, 26 Jul 2016 14:11:10 +0000 Although the use of vinasse as a waste helps replenish soil nutrients and improves the quality of the sugarcane crop, it is known that vinasse residues alter the diversity of bacteria naturally present in the soil. The actual impacts of vinasse application on the selection of bacterial taxa are not understood because no studies have addressed this phenomenon directly. Analysis of 16S rRNA gene clone sequences from four soil types showed that the soil planted with sugarcane and fertilized with vinasse has a high diversity of bacteria compared to other biomes, where Acidobacteria were the second most abundant phylum. Although the composition and structure of bacterial communities differ significantly in the four environments (Libshuff’s test), forest soils and soil planted with sugarcane without vinasse fertilizer were similar to each other because they share at least 28 OTUs related to Rhizobiales, which are important agents involved in nitrogen fixation. OTUs belonging to Actinomycetales were detected more often in the soil that had vinasse applied, indicating that these groups are more favored by this type of land management. Wellington Pine Omori, André Ferreira de Camargo, Karla Cristina Stropa Goulart, Eliana Gertrudes de Macedo Lemos, and Jackson Antônio Marcondes de Souza Copyright © 2016 Wellington Pine Omori et al. 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.