International Journal of Bacteriology The latest articles from Hindawi Publishing Corporation © 2015 , Hindawi Publishing Corporation . All rights reserved. Hyaluronidase in Clinical Isolates of Propionibacterium acnes Wed, 18 Feb 2015 06:30:27 +0000 Objectives. We sought to describe the prevalence of a hyaluronidase gene and hyaluronidase production in 197 clinical isolates of P. acnes; we assessed kinetics of hyaluronidase production in a subset of three isolates. Methods. The hyaluronidase gene was detected using polymerase chain reaction. Hyaluronidase production was detected by growing isolates on BHI agar containing 400 μg/mL hyaluronic acid and 1% albumin and flooding plates with 2 N glacial acetic acid to precipitate unbound hyaluronic acid, with a zone of clearing representing a positive phenotype. Hyaluronidase production kinetics were measured as a function of hyaluronic acid digestion over time in a liquid medium. Results. A hyaluronidase gene and hyaluronidase production were detected in 100 and 97% of P. acnes isolates, respectively. Hyaluronidase production in liquid medium was detectable after 96 hours of growth. Conclusions. Hyaluronidase production is nearly universal among P. acnes isolates. Three days appear to be required for significant hyaluronidase production in a liquid medium. Detection of hyaluronidase in tissue specimens may be a strategy to differentiate P. acnes infection from colonization when P. acnes is isolated from a clinical specimen. Harmony Tyner and Robin Patel Copyright © 2015 Harmony Tyner and Robin Patel. All rights reserved. Review on the Antimicrobial Resistance of Pathogens from Tracheal and Endotracheal Aspirates of Patients with Clinical Manifestations of Pneumonia in Bacolod City in 2013 Tue, 03 Feb 2015 06:48:59 +0000 Microbiological content specifically bacterial and fungal etiologies from tracheal aspirates in a tertiary hospital in Bacolod City was reviewed for baseline information. A total of 130 tracheal aspirates were subjected for culture to isolate and identify the pathogen and determine their susceptibilities to various antibiotics. Productions of certain enzymes responsible for antibiotic resistance like ESBL (Extended Spectrum Beta-Lactamase), metallo-β-lactamase, and carbapenemase were also studied. Out of 130 specimens, 69.23% were found to be positive for the presence of microorganisms. Most infections were from male patients aging 60 years and above, confined at the Intensive Care Units (ICU). Pseudomonas aeruginosa and Klebsiella pneumoniae were found to be the most frequent bacterial isolates and non-Candida albicans for fungal isolates, respectively. Among the various antibiotics tested, most isolates were found to be resistant to third generation cephalosporins and penicillins, but susceptible to aminoglycoside Amikacin. On the other hand, production of ESBL and carbapenemase was found to be common among members of Enterobacteriaceae especially K. pneumoniae. Alain C. Juayang, Dominador G. Maestral Jr., Gemma B. de los Reyes, Michael Angelo D. Acosido, and Christine T. Gallega Copyright © 2015 Alain C. Juayang et al. All rights reserved. Highly Sensitive Loop-Mediated Isothermal Amplification for the Detection of Leptospira Tue, 27 Jan 2015 09:21:09 +0000 Leptospirosis is a worldwide zoonosis caused by an infection with the pathogenic species of Leptospira. We have developed a loop-mediated isothermal amplification (LAMP) assay to detect the DNA of Leptospira spp. Six sets of primers targeting the gene of the subsurface protein, lipL32, were evaluated for their detection sensitivity. The best primer set detected less than 25 copies of lipL32 per reaction of both plasmid DNA template and purified leptospiral genomic DNA. By combining primers targeting lipL32 with the previously published primer set targeting lipL41, the sensitivity of the assay was improved to 12 copies of L. interrogans. The specificity of the LAMP assay was evaluated by using the genomic DNA from other clinically encountered bacterial species such as different strains of Orientia tsutsugamushi, Rickettsia typhi, Rickettsia conorii, Rickettsia rickettsii, Coxiella burnetii, and Bartonella bacilliformis. These genomic DNA samples were all negative in our LAMP assay. The sensitivity of the LAMP assay was very similar to that of quantitative real time PCR. Several detection methods for the amplified product of LAMP assay were performed to demonstrate the simplicity of the assay. In summary, our results have suggested that this assay is rapid, robust, and easy to perform and has the potential to be used in endemic locations. Hua-Wei Chen, Giulia Weissenberger, Erin Atkins, Chien-Chung Chao, Yupin Suputtamongkol, and Wei-Mei Ching Copyright © 2015 Hua-Wei Chen et al. All rights reserved. Inhibition of Quorum Sensing-Controlled Virulence Factors and Biofilm Formation in Pseudomonas aeruginosa by Culture Extract from Novel Bacterial Species of Paenibacillus Using a Rat Model of Chronic Lung Infection Sun, 11 Jan 2015 11:37:59 +0000 Quorum sensing (QS) is a key regulator of virulence factors and biofilm formation in Gram-negative bacteria such as Pseudomonas aeruginosa. Microorganisms that inhabit soil are of strategic importance in the discovery of compounds with anti-QS properties. The objective of the study was to test the culture extract of a taxonomically novel species of Paenibacillus strain 139SI for its inhibitory effects on the QS-controlled virulence factors and biofilm formation of Pseudomonas aeruginosa both in vitro and in vivo. The Paenibacillus sp. culture extract was used to test its anti-QS effects on the LasA protease, LasB elastase, pyoverdin production, and biofilm formation of P. aeruginosa as well as evaluate its therapeutic effects on lung bacteriology, pathology, hematological profile, and serum antibody responses of experimental animals in a rat model of chronic lung infection. Results showed significant decrease in the activities of QS-controlled LasA protease, LasB elastase pyoverdin, and biofilm formation of P. aeruginosa caused by the culture extract. Moreover, the extract significantly prolonged the survival times of rats and facilitated the clearance of biofilm infections from infected lungs. In conclusion, the antiquorum sensing effects of culture extract from a novel species of Paenibacillus provide new insights to combat biofilm-associated infections. Saad Musbah Alasil, Rahmat Omar, Salmah Ismail, and Mohd Yasim Yusof Copyright © 2015 Saad Musbah Alasil et al. All rights reserved. A Rapid and Sensitive Diagnostic Screening Assay for Detection of Mycobacteria Including Mycobacterium tuberculosis Directly from Sputum without Extraction Tue, 06 Jan 2015 07:16:27 +0000 We report a novel approach utilising a real-time PCR screening assay targeting a 53 bp tandemly repeated element present at various loci within the Mycobacterium tuberculosis (Mtb) genome. Positive samples were identified within a discriminatory melting curve range of 90–94°C, with results obtained in under one hour directly from decontaminated sputum samples without extraction. A panel of 89 smear-positive sputa were used for analytical validation of the assay with 100% concordance, with sensitivity matching that of culture. Cross reactivity was detected within a narrow range of mycobacteria other than tuberculosis (MOTT) (five sputa, three in silico), with the highest sensitivity within M. avium complex (MAC). A year-long head to head evaluation of the test with the GeneXpert platform was carried out with 104 consecutive samples at the Royal Free Hospital, UK. Receiver operating characteristics (ROC) analysis of the data revealed that the two tests are approximately equivalent in sensitivity, with the area under the curve being 0.85 and 0.80 for the GeneXpert and our assay, respectively, indicating that the test would be a cost effective screen prior to GeneXpert testing. Lisa Jane Cross, Catherine Anscombe, Timothy D. McHugh, Ibrahim Abubakar, Robert John Shorten, Nicola Thorne, and Cath Arnold Copyright © 2015 Lisa Jane Cross et al. All rights reserved. Study of the Antibacterial Efficacy of Bainiku-Ekisu against Pathogens Tue, 28 Oct 2014 06:38:14 +0000 The research was undertaken to determine the bacteriostatic effects of the concentrate of Japanese apricot juice (bainiku-ekisu), which is a popular health food in Taiwan and Japan, on Enterococcus faecalis ATCC 29212, Staphylococcus aureus ATCC 25923, and Escherichia coli ATCC 25922. The results show that E. faecalis, S. aureus, and E. coli could be killed or inhibited by bainiku-ekisu at concentrations between 1.0 and 10.0 mg/mL. The minimum inhibitory concentration (MIC) was 1 mg/mL for all strains, and the minimum bactericidal concentrations (MBCs) were 5, 2.5, and 2.5 mg/mL for E. faecalis, S. aureus, and E. coli, respectively. Using the growth rate to calculate the MICs and MBCs, the MICs were 1.55, 1.43, and 0.97 mg/mL, and the MBCs were 2.59, 2.63, and 2.25 mg/mL for E. faecalis, S. aureus, and E. coli, respectively. According to the D values, E. faecalis and S. aureus exhibited lower resistance than E. coli at lower bainiku-ekisu concentrations (1.0 and 2.5 mg/mL), and the resistance of these two pathogens was better than that of E. coli at higher bainiku-ekisu concentrations (5.0 and 10.0 mg/mL). The Z values of the E. faecalis, S. aureus, and E. coli strains were 3.47, 4.93, and 11.62 mg/mL, respectively. Deng-Jye Yang, Hsin-Yi Chen, and Shih-Chuan Liu Copyright © 2014 Deng-Jye Yang et al. All rights reserved. Activity of Aristolochia bracteolata against Moraxella catarrhalis Sun, 28 Sep 2014 09:11:22 +0000 A bioassay-guided fractionation of methanol extract of Aristolochia bracteolata whole plant was carried out in order to evaluate its antimicrobial activity and to identify the active compounds in this extract. Antibacterial and antifungal activities of methanol extract against gram-positive, gram-negative, and fungal strains were investigated by the agar disk diffusion method. Among the strains tested, Moraxella catarrhalis and sea urchin-derived Bacillus sp. showed the highest sensitivity towards the methanol extract and hence they are used as test organisms for the bioassay-guided fractionation. From this extract, aristolochic acid 1 (AA-1) has been isolated and has showed the greatest antibacterial activity against both standard strain and clinical isolates of Moraxella catarrhalis with equal minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values of 25 and 50 μg/mL. Modification of the AA-1 to AA-1 methyl ester completely abolished the antibacterial activity of the compound and the piperonylic acid moiety of AA-1 which suggested that the coexistence of phenanthrene ring and free carboxylic acid is essential for AA-1 antibacterial activity. Malik Suliman Mohamed, Mona Timan Idriss, Amgad I. M. Khedr, Haidar Abd AlGadir, Satoshi Takeshita, Mohammad Monir Shah, Yoshio Ichinose, and Toshihide Maki Copyright © 2014 Malik Suliman Mohamed et al. All rights reserved. The Frequency of Methicillin-Resistant Staphylococcus aureus and Coagulase Gene Polymorphism in Egypt Wed, 24 Sep 2014 00:00:00 +0000 The current study aimed to use Coagulase gene polymorphism to identify methicillin-resistant Staphylococcus aureus (MRSA) subtypes isolated from nasal carriers in Minia governorate, Egypt, evaluate the efficiency of these methods in discriminating variable strains, and compare these subtypes with antibiotypes. A total of 400 specimens were collected from nasal carriers in Minia governorate, Egypt, between March 2012 and April 2013. Fifty-eight strains (14.5%) were isolated and identified by standard microbiological methods as MRSA. The identified isolates were tested by Coagulase gene RFLP typing. Out of 58 MRSA isolates 15 coa types were classified, and the amplification products showed multiple bands (1, 2, 3, 4, 5, and 8 bands). Coagulase gene PCR-RFLPs exhibited 10 patterns that ranged from 1 to 8 fragments with AluI digestion. Antimicrobial susceptibility testing with a panel of 8 antimicrobial agents showed 6 different antibiotypes. Antibiotype 1 was the most common phenotype with 82.7%. The results have demonstrated that many new variants of the coa gene are present in Minia, Egypt, different from those reported in the previous studies. So surveillance of MRSA should be continued. Hend M. Abdulghany and Rasha M. Khairy Copyright © 2014 Hend M. Abdulghany and Rasha M. Khairy. All rights reserved. Mix Infections of Helicobacter pylori: A Major Risk Factor Affecting Genotyping Studies Thu, 05 Jun 2014 15:55:26 +0000 Amin Talebi Bezmin Abadi Copyright © 2014 Amin Talebi Bezmin Abadi. All rights reserved. Antibacterial Attributes of Apigenin, Isolated from Portulaca oleracea L. Tue, 13 May 2014 11:03:21 +0000 The flavonoid apigenin was isolated from aerial part of P. oleracea L. The dried sample of plant was powdered and subjected to soxhlet extractor by adding 80 mL of ethanol : water (70 : 30). The extract was centrifuged at 11000 rpm for 30 min; supernatant was taken for further use. The fraction was concentrated and subjected to PTLC. The value of isolated apigenin was calculated (0.82). Purified material was also subjected to its IR spectra, LC-MS, NMR, and HPLC for structural elucidation. The apigenin so-obtained was subjected to antibacterial activity on five pathogenic bacterial strains like Pseudomonas aeruginosa, Salmonella typhimurium, Proteus mirabilis, Klebsiella pneumoniae and Enterobacter aerogenes; among all the bacterial strains, Salmonella typhimurium and Proteus mirabilis have shown maximum diameter of inhibition zone for flavonoid and remaining bacterial strains have shown moderate diameter of inhibition zone when compared with control values and , respectively. The minimum inhibitory concentration (MIC) of the flavonoid isolated from P. oleracea L. was tested at the concentration ranging from undiluted sample to 10 mg per mL of concentration. The minimum inhibition concentration (MIC) for the flavonoid for all tested bacterial strains was found to be >4 mg per mL. Hence, the apigenin has antibacterial property and can be used to develop antibacterial drugs. Hanumantappa B. Nayaka, Ramesh L. Londonkar, Madire K. Umesh, and Asha Tukappa Copyright © 2014 Hanumantappa B. Nayaka et al. All rights reserved. Evaluation of the Pattern of EPIYA Motifs in the Helicobacter pylori cagA Gene of Patients with Gastritis and Gastric Adenocarcinoma from the Brazilian Amazon Region Thu, 24 Apr 2014 16:47:38 +0000 The Helicobacter pylori is associated with the development of different diseases. The clinical outcome of infection may be associated with the cagA bacterial genotype. The aim of this study was to determine the EPIYA patterns of strains isolated from patients with gastritis and gastric adenocarcinoma and correlate these patterns with the histopathological features. Gastric biopsy samples were selected from 384 patients infected with H. pylori, including 194 with chronic gastritis and 190 with gastric adenocarcinoma. The presence of the cagA gene and the EPIYA motif was determined by PCR. The cagA gene was more prevalent in patients with gastric cancer and was associated with a higher degree of inflammation, neutrophil activity, and development of intestinal metaplasia. The number of EPIYA-C repeats showed a significant association with an increased risk of gastric carcinoma (OR = 3.79, 95% CI = 1.92–7.46, and ). A larger number of EPIYA-C motifs were also associated with intestinal metaplasia. In the present study, infection with H. pylori strains harboring more than one EPIYA-C motif in the cagA gene was associated with the development of intestinal metaplasia and gastric adenocarcinoma but not with neutrophil activity or degree of inflammation. Adenielson Vilar e Silva, Mario Ribeiro da Silva Junior, Ruth Maria Dias Ferreira Vinagre, Kemper Nunes Santos, Renata Aparecida Andrade da Costa, Amanda Alves Fecury, Juarez Antônio Simões Quaresma, and Luisa Caricio Martins Copyright © 2014 Adenielson Vilar e Silva et al. All rights reserved. Detection of Q Fever Specific Antibodies Using Recombinant Antigen in ELISA with Peroxidase Based Signal Amplification Wed, 12 Mar 2014 11:05:01 +0000 Currently, the accepted method for Q fever serodiagnosis is indirect immunofluorescent antibody assay (IFA) using the whole cell antigen. In this study, we prepared the recombinant antigen of the 27-kDa outer membrane protein (Com1) which has been shown to be recognized by Q fever patient sera. The performance of recombinant Com1 was evaluated in ELISA by IFA confirmed serum samples. Due to the low titers of IgG and IgM in Q fever patients, the standard ELISA signals were further amplified by using biotinylated anti-human IgG or IgM plus streptavidin-HRP polymer. The modified ELISA can detect 88% (29 out of 33) of Q fever patient sera collected from Marines deployed to Iraq. Less than 5% (5 out of 156) of the sera from patients with other febrile diseases reacted with the Com1. These results suggest that the modified ELISA using Com1 may have the potential to improve the detection of Q fever specific antibodies. Hua-Wei Chen, Zhiwen Zhang, Erin Glennon, and Wei-Mei Ching Copyright © 2014 Hua-Wei Chen et al. All rights reserved. Changing Trends in Prevalence and Antibiotics Resistance of Uropathogens in Patients Attending the Gondar University Hospital, Northwest Ethiopia Mon, 10 Mar 2014 12:25:40 +0000 Background. In most hospitals of developing countries, urinary tract infections are treated empirically because of lack of culture facilities. This leads to emergence of multiresistant uropathogens. Culturing and drug susceptibility testing are essential to guide therapy. Objectives. To assess changing prevalence and resistance pattern of uropathogens to commonly used antibiotics in a two-year study period. Methods. Urine specimens were collected and cultured. Uropathogens were identified by standard methods and tested for antibiotics resistance. Data were analyzed using SPSS version 16 statistical sofware. P value < 0.05 was considered statistically significant. Results. The commonest isolates in both the previous and present studies were E. coli, Klebsiella, CoNS, S. aureus, Proteus, and Citrobacter species. Previous isolates of Enterobacteriaceae were 100% sensitive to ciprofloxacin, whereas present isolates developed 31% to 60% resistance to it. Previous isolates were less resistant to gentamycin than the present ones. Multiresistance isolates were predominant in present study than previous ones. Conclusion. E. coli was predominant in the two study periods. Present isolates were more resistant than previous ones. Some previous isolates were 100% sensitive to ciprofloxacin, whereas present isolates were increasingly resistant. Ciprofloxacin and gentamicin have been recommended for empiric treatment of urinary tract infections. Moges Tiruneh, Sisay Yifru, Mucheye Gizachew, Kassie Molla, Yeshambel Belyhun, Feleke Moges, and Mengistu Endris Copyright © 2014 Moges Tiruneh et al. All rights reserved. In Vitro Screening for Abiotic Stress Tolerance in Potent Biocontrol and Plant Growth Promoting Strains of Pseudomonas and Bacillus spp. Thu, 06 Mar 2014 15:57:52 +0000 Plant growth promoting rhizobacteria (PGPR) has been identified as a group of microbes that are used for plant growth enhancement and biocontrol for management of plant diseases. The inconsistency in performance of these bacteria from laboratory to field conditions is compounded due to the prevailing abiotic stresses in the field. Therefore, selection of bacterial strains with tolerance to abiotic stresses would benefit the end-user by successful establishment of the strain for showing desired effects. In this study we attempted to isolate and identify strains of Bacillus and Pseudomonas spp. with stress tolerance and proven ability to inhibit the growth of potential phytopathogenic fungi. Screening of bacterial strains for high temperature (50°C), salinity (7% NaCl), and drought (−1.2 MPa) showed that stress tolerance was pronounced less in Pseudomonas isolates than in Bacillus strains. The reason behind this could be the formation of endospores by Bacillus isolates. Tolerance to drought was high in Pseudomonas strains than the other two stresses. Three strains, P8, P20 and P21 showed both salinity and temperature tolerance. P59 strain possessed promising antagonistic activity and drought tolerance. The magnitude of antagonism shown by Bacillus isolates was also higher when compared to Pseudomonas strains. To conclude, identification of microbial candidate strains with stress tolerance and other added characteristic features would help the end-user obtain the desired beneficial effects. G. Praveen Kumar, S. K. Mir Hassan Ahmed, Suseelendra Desai, E. Leo Daniel Amalraj, and Abdul Rasul Copyright © 2014 G. Praveen Kumar et al. All rights reserved. Mixture of Sodium Hypochlorite and Hydrogen Peroxide on Adhered Aeromonas hydrophila to Solid Substrate in Water: Impact of Concentration and Assessment of the Synergistic Effect Mon, 03 Mar 2014 16:43:17 +0000 The synergistic effects of the combined treatments of NaOCl and H2O2 on the elimination of A. hydrophila adhered to polythene under static and dynamic conditions were evaluated. The concentrations 0.1, 0.2, and 0.3 NaOCl and 0.5, 1, and 1.5 H2O2 were used. The contact periods were 180, 360, 540, and 720 minutes. The abundance of cells adhered reached 2.47 and 2.27 units (log (CFU/cm²)), respectively, under static and dynamic conditions after action of the mixture of disinfectants, whereas it reached 2.41 and 3.39 units (log (CFU/cm²)) after action of NaOCl and H2O2 alone, respectively. Increase in the incubation period resulted in a significant decrease in the abundance of cells adhered when the mixture of 0.3 NaOCl and 1.5 H2O2 was used (). For each cell growth phase, there was a significant difference amongst the mean densities of cells adhered after action of the mixture of disinfectants (). Although the Freundlich isotherm parameters relatively varied from one experimental condition to another, the value registered in the exponential growth phase was relatively higher in static state than in dynamic regime; cells adhered under dynamic condition seem more sensitive to the synergistic action than those adhered under static condition. Chrétien Lontsi Djimeli, Antoine Tamsa Arfao, Olive V. Noah Ewoti, Mireille Ebiane Nougang, Marlyse L. Moungang, Geneviève Bricheux, Moïse Nola, and Télesphore Sime-Ngando Copyright © 2014 Chrétien Lontsi Djimeli et al. All rights reserved. Antibiofilm Activity of Manuka Honey in Combination with Antibiotics Wed, 26 Feb 2014 12:01:05 +0000 We assessed the in vitro activity of Manuka honey against biofilm bacteria in combination with antibiotics and visualized the effect of Manuka honey on bacterial biofilms using scanning electron microscopy. The fractional biofilm eradication concentration () index for vancomycin plus Manuka honey against S. aureus IDRL-4284 biofilms was 0.34, indicating a synergistic interaction. The index for gentamicin plus Manuka honey against P. aeruginosa PAO1 biofilms was 0.78–0.82, indicating an additive interaction. Scanning electron microscopy of S. aureus IDRL-4284 and P. aeruginosa PAO1 biofilms exposed to Manuka honey and artificial honey containing the same sugar composition as Manuka honey showed that the former had more pronounced effects than the latter on both S. aureus and P. aeruginosa biofilms. Visualized effects included distorted cell morphologies for both bacteria and a decrease in P. aeruginosa extracellular matrix. In conclusion, Manuka honey has a synergistic interaction with vancomycin against S. aureus biofilms and an additive interaction with gentamicin against P. aeruginosa biofilms. Michelle E. M. Campeau and Robin Patel Copyright © 2014 Michelle E. M. Campeau and Robin Patel. All rights reserved. Phylogenetic Framework and Biosurfactant Gene Expression Analysis of Marine Bacillus spp. of Eastern Coastal Plain of Tamil Nadu Wed, 12 Feb 2014 08:23:51 +0000 The present study emphasizes the diversity assessment of marine Bacillus species with special reference to biosurfactant production, respective gene expression, and discrimination among Bacillus licheniformis and Bacillus subtilis. Among the 200 individual species of eastern coastal plain of Tamil Nadu screened, five biosurfactant producing potential bacterial species with entirely different morphology were selected. Biochemical and 16S rRNA gene sequence analysis suggested that all the said five species belong to Bacillus genera but differ in species levels. Biosurfactant of all the five species fluctuates in greater levels with respect to activity as well as to constituents but showed partial similarity to the commercially available surfactin. The expression of srf gene was realized in all of the five species. However, the sfp gene expression was observed only in three species. In conclusion, both B. licheniformis and B. subtilis demonstrate srf gene; nevertheless, sfp gene was expressed only by Bacillus subtilis. Sreethar Swaathy, Varadharajan Kavitha, Arockiasamy Sahaya Pravin, Ganesan Sekaran, Asit Baran Mandal, and Arumugam Gnanamani Copyright © 2014 Sreethar Swaathy et al. All rights reserved. The Assessment of Viability of M. Tuberculosis after Exposure to CPC Using Different Methods Tue, 28 Jan 2014 07:25:33 +0000 Settings. National Institute for Research in Tuberculosis, Chennai. Objective. To assess the proportion of metabolically active cells of Mycobacterium tuberculosis after exposed to CPC using FDA-EB vital staining and viable counts on LJ medium. Mycolic acid content in M. tuberculosis after exposure to CPC was estimated using HPLC. Methods. Clinical isolates of M. tuberculosis and standard reference strain M. tuberculosis H37Rv were used for FDA-EB, viable count, and HPLC. Results. FDA/EB consistently stained 70–90% of log phase cells as green and the remaining cells as red-orange. After CPC treatment, 65–70% of the cells stained red-orange. The viability counts were comparable to 0-day controls. Synthesis of mycolic acids in mycobacteria was reduced when exposed to CPC using HPLC due to the decreased metabolic activity of the organisms. Conclusion. The cells are metabolically inactive during storage with CPC but these cells grew well on LJ medium after removal of CPC. The viability of M. tuberculosis was maintained in CPC with minimal reduction. Mycolic acid content was reduced if the cells of M. tuberculosis were treated with CPC for 7 days. All the above findings provide yet another evidence for the damage of cell wall of M. tuberculosis. Gomathi Sekar, R. Lakshmi, and N. Selvakumar Copyright © 2014 Gomathi Sekar et al. All rights reserved. Bacterial Biodegradation of Crude Oil Using Local Isolates Mon, 20 Jan 2014 06:44:45 +0000 An experimental study was undertaken to assess the efficiency of Pseudomonas aeruginosa, Bacillus subtilis, and Acinetobacter lwoffi isolated from petroleum contaminated water and soil samples to degrade crude oil, separately and in a mixed bacterial consortium. Capillary gas chromatography was used for testing the effect of those bacterial species on the biodegradation of crude oil. Individual bacterial cultures showed less growth and degradation than did the mixed bacterial consortium. At temperature 22°C, the mixed bacterial consortium degraded a maximum of 88.5% of Egyptian crude oil after 28 days of incubation. This was followed by 77.8% by Pseudomonas aeruginosa, 76.7% by Bacillus subtilis, and 74.3% by Acinetobacter lwoffi. The results demonstrated that the selected bacterial isolates could be effective in biodegradation of oil spills individually and showed better biodegradation abilities when they are used together in mixed consortium. Raed S. Al-Wasify and Shimaa R. Hamed Copyright © 2014 Raed S. Al-Wasify and Shimaa R. Hamed. All rights reserved. Isolation and Antimicrobial Susceptibility Patterns of Campylobacter Species among Diarrheic Children at Jimma, Ethiopia Sun, 12 Jan 2014 00:00:00 +0000 Introduction. Campylobacter is one of the leading bacterial causes of food-borne disease. The prevalence of Campylobacter species resistant to antimicrobial agents is increasing. This study is intended to determine prevalence and antimicrobial susceptibility patterns of Campylobacter species among under-five children with diarrhea. Methodology. A cross-sectional study was conducted among 227 under-five children with diarrhea from July to October 2012 at Jimma town. Isolation and identification of Campylobacter species were performed using standard bacteriological techniques. Antimicrobial susceptibility test was performed following standard protocol. Chi-square and Fisher’s exact tests were used for analysis. Results. From 227 under-five children, 16.7% were positive for Campylobacter spp.; isolates, C. jejuni, C. coli, and C. lari, accounted for 71.1%, 21.1%, and 7.9%, respectively. Higher rate of resistance was observed to ampicillin 76.3%, trimethoprim-sulfamethoxazole (68.4%), tetracycline (39.5%), chloramphenicol (31.6%), clindamycin (26.3%), and doxycycline (23.7%). Erythromycin, ciprofloxacin, gentamicin, norfloxacin, and nalidixic acid were effective for more than 80% of the isolates. Multiple drug resistance was observed among 78.9% of all the three spp. Conclusions. Isolation rate of Campylobacter spp. was high. C. lari was reported for the first time at this study area. Higher rate of resistance was observed to the commonly used drugs. Belay Tafa, Tsegaye Sewunet, Haimanot Tassew, and Daniel Asrat Copyright © 2014 Belay Tafa et al. All rights reserved. New Medium for Pharmaceutical Grade Arthrospira Sat, 28 Dec 2013 12:38:48 +0000 The aim of this study is to produce a pharmaceutical grade single cell product of Arthrospira from a mixed culture. We have designed a medium derived from a combination between George’s and Zarrouk’s media. Our new medium has the ability to inhibit different forms of cyanobacterium and microalgae except the Chlorella. The medium and the cultivation conditions have been investigated to map the points where only Arthrospira could survive. For that, a mixed culture of pure Chlorella and Arthrospira (~90 : 10) has been used to develop the best medium composition that can lead to the enrichment of the Arthrospira growth and the inhibition of the Chlorella growth. To enable better control and to study its growth, an 80 l photobioreactor has been used. We have used high saline (2xA-St) medium which has been followed by in fermentor reducing its concentration to 1.5x. The investigation proves that Chlorella has completely disappeared. A method and a new saline medium have been established using a photobioreactor for in fermentor production of single cell Arthrospira. Such method enables the production of pure pharmaceutical grade Arthrospira for medicinal and pharmaceutical applications or as a single cell protein. Amro A. Amara and Alexander Steinbüchel Copyright © 2013 Amro A. Amara and Alexander Steinbüchel. All rights reserved. Expression, Purification, and Functional Characterization of Atypical Xenocin, Its Immunity Protein, and Their Domains from Xenorhabdus nematophila Wed, 18 Dec 2013 09:49:00 +0000 Xenorhabdus nematophila, a gram-negative bacterium belonging to the family Enterobacteriaceae is a natural symbiont of a soil nematode from the family Steinernematidae. In this study cloning, expression, and purification of broad range iron regulated multidomain bacteriocin called xenocin from X. nematophila (66 kDa, encoded by xcinA gene) and its multidomain immunity protein (42 kDa, encoded by ximB gene) have been done. xcinA-ximB (N′ terminal 270 bp), translocation, and translocation-receptor domain of xcinA, ximB, and its hemolysin domain were cloned, expressed, and purified by single step Ni-NTA chromatography under native conditions. In the functional characterization, neutralization of xcinA toxicity by immunity domain of ximB gene was determined by endogenous assay. Exogenous toxic assays results showed that only the purified recombinant xenocin-immunity domain (10 kDa) protein complex had toxic activity. Atypical cognate immunity protein (42 kDa) of xenocin was fusion of immunity domain (10 kDa) and hemolysin domain (32 kDa). In silico analysis of immunity protein revealed its similarity with hemolysin and purine NTPase like proteins. Hemolytic activity was not observed in immunity protein or in its various domains; however, full-length immunity protein lacking Walker motif showed ATPase activity. Finally, using circular dichroism performed secondary structural analyses of all the recombinant proteins/protein complexes. Jitendra Singh Rathore Copyright © 2013 Jitendra Singh Rathore. All rights reserved. Modulation of Bacterial Multidrug Resistance Efflux Pumps of the Major Facilitator Superfamily Thu, 05 Dec 2013 18:45:35 +0000 Bacterial infections pose a serious public health concern, especially when an infectious disease has a multidrug resistant causative agent. Such multidrug resistant bacteria can compromise the clinical utility of major chemotherapeutic antimicrobial agents. Drug and multidrug resistant bacteria harbor several distinct molecular mechanisms for resistance. Bacterial antimicrobial agent efflux pumps represent a major mechanism of clinical resistance. The major facilitator superfamily (MFS) is one of the largest groups of solute transporters to date and includes a significant number of bacterial drug and multidrug efflux pumps. We review recent work on the modulation of multidrug efflux pumps, paying special attention to those transporters belonging primarily to the MFS. Sanath Kumar, Mun Mun Mukherjee, and Manuel F. Varela Copyright © 2013 Sanath Kumar et al. All rights reserved. Inflammatory Responses to Salmonella Infections Are Serotype-Specific Mon, 16 Sep 2013 08:44:31 +0000 The main purpose of this study was to investigate the profile of inflammatory response in patients with acute salmonellosis caused by two serotypes of Salmonella enterica, S. Enteritidis and S. Typhimurium, as well as in convalescent patients with previous acute disease caused by S. Enteritidis. Patients with acute disease showed significantly elevated levels of IL-1β, IL-17, IL-10, and calprotectin compared to healthy control subjects. In convalescent patients, these markers were also significantly elevated, with the exception of IL-1β. Multivariate statistical analyses with the use of these variables produced models with a good predictive accuracy resulting in excellent separation of the diseased and healthy cohorts studied. Overall, the results suggest that the profile of inflammatory response in this disease is determined, to a significant degree, by the serotype of Salmonella, and the profile of certain cytokines and calprotectin remains abnormal for a number of months following the acute disease stage. Zhanna Ktsoyan, Karine Ghazaryan, Gayane Manukyan, Anush Martirosyan, Armine Mnatsakanyan, Karine Arakelova, Zaruhi Gevorgyan, Anahit Sedrakyan, Ara Asoyan, Anna Boyajyan, and Rustam Aminov Copyright © 2013 Zhanna Ktsoyan et al. All rights reserved. Survival and Growth of Vibrio cholerae, Escherichia coli, and Salmonella Spp. in Well Water Used for Drinking Purposes in Garoua (North Cameroon) Wed, 07 Aug 2013 08:09:25 +0000 The ability of strains of faecal bacteria (Vibrio cholerae, Escherichia coli ATCC 25922, and four strains of Salmonella isolated, resp., from well water, pig, poultry, and human urine in Garoua) to survive or grow in well water microcosms was compared. Water samples were obtained from two wells in Garoua (north Cameroun). Autoclaving at 121°C for 15 min and filtration through 0.2 µm filter were used to make microcosms. Microcosms were constituted of unfiltered-autoclaved, filtered-nonautoclaved, and filtered-autoclaved well waters. Bacterial strains were inoculated at initial cell concentration of 3 Log10CFU/mL. All strains were able to survive/grow in used microcosms, and a maximal concentration of 5.61 Log10CFU/mL was observed. Survival abilities were strain and microcosm dependent. The declines were more pronounced in filtered-nonautoclaved water than in the other microcosms. E. coli and Salmonella sp. (poultry strain) lowered to undetectable levels (<1 Log10CFU/mL) after two days of water storage. V. cholera decreased over time, but surviving cells persisted for longer period in filtered-nonautoclaved water from well W1 (1.91 Log10CFU/mL) and well W2 (2.09 Log10CFU/mL). Competition for nutrients and/or thermolabile antimicrobial substances synthesized by “ultramicrocells” or by the autochthonous bacteria retained by the filter might affect the bacterial survival. Moussa Djaouda, Bouba Gaké, Daniel Ebang Menye, Serge Hubert Zébazé Togouet, Moïse Nola, and Thomas Njiné Copyright © 2013 Moussa Djaouda et al. All rights reserved. Physiological Properties and Salmonella Growth Inhibition of Probiotic Bacillus Strains Isolated from Environmental and Poultry Sources Sun, 26 May 2013 14:49:27 +0000 The objective of the present study was to describe the physiological properties of seven potential probiotic strains of Bacillus spp. Isolates were characterized morphologically, biochemically, and by 16S rRNA sequence analyses for identification. Tolerance to acidic pH, high osmotic concentrations of NaCl, and bile salts were tested. Isolates were also evaluated for their ability to metabolize different carbohydrates sources. The antimicrobial sensitivity profiles were determined. Inhibition of gastrointestinal Salmonella colonization in an avian model was also evaluated. Five strains of Bacillus were tolerant to acidic conditions (pH 2.0) and all strains were tolerant to a high osmotic pressure (NaCl at 6.5%). Moreover, all strains were able to tolerate concentration of 0.037% bile salts after 24 h of incubation. Three strains were able to significantly reduce Salmonella Typhimurium levels in the crop and in the ceca of broiler-type chickens. Among the 12 antibiotics tested for antibiotic resistance, all strains were resistant to bacitracin and susceptible to gentamycin, neomycin, ormethoprim, triple sulfa, and spectinomycin. Bacterial spore formers have been shown to prevent gastrointestinal diseases in animals and humans. The results obtained in this study show important characteristics to be evaluated when selecting Bacillus spp. candidates to be used as probiotics. Anita Menconi, Marion J. Morgan, Neil R. Pumford, Billy M. Hargis, and Guillermo Tellez Copyright © 2013 Anita Menconi et al. All rights reserved. Decreased C3 Activation by the devR Gene-Disrupted Mycobacterium tuberculosis Strain in Comparison to the Wild-Type Strain Sat, 18 May 2013 09:47:06 +0000 Activation of the complement component C3 is an important step in the complement cascade, contributing to inflammatory mechanisms. Considerable research on gene-disrupted mycobacterial strains using animal models of tuberculosis infection has reported the roles of some of the mycobacterial genes during tuberculosis infection. The aim of the present study was to assess the pattern of complement activation by the devR gene-disrupted Mycobacterium tuberculosis H37Rv strain and compare with that by its wild-type strain. In vitro complement activation at the level of C3 by the gene-disrupted strain, its complemented strain, and wild-type strain was performed using solid-phase ELISA. It was observed that the ability of devR gene-disrupted M. tuberculosis H37Rv to activate C3 was significantly reduced in comparison to its wild-type strain (). In addition, C3 activation by the complemented devR mutant strain was almost similar to that of the wild strain, which indicated that the reduced ability to activate C3 could potentially be due to the deletion of devR gene. These findings indicate that the gene devR probably aids in complement activation and contributes to the inflammatory processes during tuberculosis infection. V. Narayan Rao, S. Manivannan, J. S. Tyagi, and V. D. Ramanathan Copyright © 2013 V. Narayan Rao et al. All rights reserved. Detection of Bordetella pertussis from Clinical Samples by Culture and End-Point PCR in Malaysian Patients Tue, 07 May 2013 09:03:26 +0000 Pertussis or whooping cough is a highly infectious respiratory disease caused by Bordetella pertussis. In vaccinating countries, infants, adolescents, and adults are relevant patients groups. A total of 707 clinical specimens were received from major hospitals in Malaysia in year 2011. These specimens were cultured on Regan-Lowe charcoal agar and subjected to end-point PCR, which amplified the repetitive insertion sequence IS481 and pertussis toxin promoter gene. Out of these specimens, 275 were positive: 4 by culture only, 6 by both end-point PCR and culture, and 265 by end-point PCR only. The majority of the positive cases were from ≤3 months old patients (77.1%) (). There was no significant association between type of samples collected and end-point PCR results (). Our study showed that the end-point PCR technique was able to pick up more positive cases compared to culture method. Tan Xue Ting, Rohaidah Hashim, Norazah Ahmad, and Khairul Hafizi Abdullah Copyright © 2013 Tan Xue Ting et al. All rights reserved. Rapid PCR Detection of Mycoplasma hominis, Ureaplasma urealyticum, and Ureaplasma parvum Mon, 11 Mar 2013 09:14:04 +0000 Objective. We compared laboratory developed real-time PCR assays for detection of Mycoplasma hominis and for detection and differentiation of Ureaplasma urealyticum and parvum to culture using genitourinary specimens submitted for M. hominis and Ureaplasma culture. Methods. 283 genitourinary specimens received in the clinical bacteriology laboratory for M. hominis and Ureaplasma species culture were evaluated. Nucleic acids were extracted using the Total Nucleic Acid Kit on the MagNA Pure 2.0. 5 μL of the extracts were combined with 15 μL of each of the two master mixes. Assays were performed on the LightCycler 480 II system. Culture was performed using routine methods. Results.  M. hominis PCR detected 38/42 M. hominis culture-positive specimens, as well as 2 that were culture negative (sensitivity, 90.5%; specificity, 99.2%). Ureaplasma PCR detected 139/144 Ureaplasma culture-positive specimens, as well as 9 that were culture negative (sensitivity, 96.5%; specificity, 93.6%). Of the specimens that tested positive for Ureaplasma species, U. urealyticum alone was detected in 33, U. parvum alone in 109, and both in 6. Conclusion. The described PCR assays are rapid alternatives to culture for detection of M. hominis and Ureaplasma species, and, unlike culture, the Ureaplasma assay easily distinguishes U. urealyticum from parvum. Scott A. Cunningham, Jayawant N. Mandrekar, Jon E. Rosenblatt, and Robin Patel Copyright © 2013 Scott A. Cunningham et al. All rights reserved.