International Journal of Microbiology The latest articles from Hindawi Publishing Corporation © 2014 , Hindawi Publishing Corporation . All rights reserved. The TIR Domain Containing Locus of Enterococcus faecalis Is Predominant among Urinary Tract Infection Isolates and Downregulates Host Inflammatory Response Thu, 24 Jul 2014 12:52:27 +0000 Based on Toll/interleukin-1 receptor (TIR) domain structure homology, we detected a previously uncharacterized gene encoding for a TIR domain containing protein (Tcp) in the genome of Enterococcus faecalis. We assigned this gene the name tcpF (as in Tcp of E. faecalis). Screening of E. faecalis samples revealed that tcpF is more common in isolates from urinary tract infections (UTIs) than in human faecal flora. tcpF alleles showed moderate single nucleotide polymorphism (SNP) among UTI isolates. Infection of mouse RAW264.7 macrophages with a tcpF knock-out mutant led to elevated cytokine response compared to the isogenic wild type E. faecalis strain. In silico analysis predicted significant tertiary structure homology to the TIR domain of human TLR1 (TLR1-TIR). When transiently expressed in cultured eukaryotic cells, TcpF caused suppression of TLR2-dependent NF-κB activation suggesting for TcpF a role as a factor in E. faecalis that benefits colonization by modulating the host’s immune responses. Thomas Daniel Kraemer, Orlando Daniel Quintanar Haro, Eugen Domann, Trinad Chakraborty, and Svetlin Tchatalbachev Copyright © 2014 Thomas Daniel Kraemer et al. All rights reserved. Identification and In Vitro Antimicrobial Susceptibility of Brucella Species Isolated from Human Brucellosis Thu, 10 Jul 2014 08:44:41 +0000 Brucellosis is a world-wide zoonotic disease with a major impact on the public health. Due to the high risk of laboratory acquired infection, limited laboratory investigations were performed on this organism, including detailed identification and susceptibility study. Brucella melitensis is the commonest aetiological agent for human brucellosis in this region. The in vitro susceptibility pattern against selected antimicrobial agents was assessed using E-test. All isolates were noted to be sensitive to all the antimicrobial agents tested except for rifampicin where elevated MIC > 1 μg/mL was noted in 30 out of 41 isolates tested. Rohaidah Hashim, Norazah Ahmad, Jama’ayah Mohamed Zahidi, B. Y. Tay, Azura Mohd Noor, Sakina Zainal, Hazwani Hamzah, S. H. Hamzah, T. S. Chow, P. S. Wong, and K. N. Leong Copyright © 2014 Rohaidah Hashim et al. All rights reserved. Comparative Analysis of Protein Glycosylation Pathways in Humans and the Fungal Pathogen Candida albicans Thu, 03 Jul 2014 06:04:06 +0000 Protein glycosylation pathways are present in all kingdoms of life and are metabolic pathways found in all the life kingdoms. Despite sharing commonalities in their synthesis, glycans attached to glycoproteins have species-specific structures generated by the presence of different sets of enzymes and acceptor substrates in each organism. In this review, we present a comparative analysis of the main glycosylation pathways shared by humans and the fungal pathogen Candida albicans: -linked glycosylation, -linked mannosylation and glycosylphosphatidylinositol-anchorage. The knowledge of similarities and divergences between these metabolic pathways could help find new pharmacological targets for C. albicans infection. Iván Martínez-Duncker, Diana F. Díaz-Jímenez, and Héctor M. Mora-Montes Copyright © 2014 Iván Martínez-Duncker et al. All rights reserved. Microbiologic Spectrum and Antibiotic Susceptibility Pattern among Patients with Urinary and Respiratory Tract Infection Sun, 29 Jun 2014 00:00:00 +0000 Aim. To demonstrate the prevalence of isolated organisms in urinary/respiratory tract infections and their antibiotic susceptibilities in a tertiary care center. Methods and Material. Between January 2008 and January 2010, patients referring to the clinic of cardiology or those admitted to the cardiac wards were enrolled in this cross-sectional descriptive study. Urine and sputum sampling was done for all the patients and the specimens underwent microbiologic examination and, in case of isolation of microorganism, antibiotic disk diffusion test was performed. Results. Escherichia coli (E. coli) was the most prevalent isolated organism in-hospital and community-acquired UTIs and was highly resistant to cephalothin in all the samples followed by cotrimoxazole, and ceftriaxone. It revealed high sensitivity to imipenem, amikacin, and nitrofurantoin. Acinetobacter constituted the most prevalent organism isolated from respiratory secretions and represented the highest resistance to ceftriaxone and the greatest sensitivity to imipenem. Conclusions. E. coli and Acinetobacter remain the most common uropathogenic and respiratory organisms, respectively. However, their increasing resistance to wide-spectrum imipenem, meropenem, and vancomycin is a major concern. Maryam Sotoudeh Anvari, Mohammad Naderan, Mohammad Ali Boroumand, Saeed Shoar, Robab Bakhshi, and Morteza Naderan Copyright © 2014 Maryam Sotoudeh Anvari et al. All rights reserved. Response Mechanisms of Bacterial Degraders to Environmental Contaminants on the Level of Cell Walls and Cytoplasmic Membrane Thu, 26 Jun 2014 09:15:04 +0000 Bacterial strains living in the environment must cope with the toxic compounds originating from humans production. Surface bacterial structures, cell wall and cytoplasmic membrane, surround each bacterial cell and create selective barriers between the cell interior and the outside world. They are a first site of contact between the cell and toxic compounds. Organic pollutants are able to penetrate into cytoplasmic membrane and affect membrane physiological functions. Bacteria had to evolve adaptation mechanisms to counteract the damage originated from toxic contaminants and to prevent their accumulation in cell. This review deals with various adaptation mechanisms of bacterial cell concerning primarily the changes in cytoplasmic membrane and cell wall. Cell adaptation maintains the membrane fluidity status and ratio between bilayer/nonbilayer phospholipids as well as the efflux of toxic compounds, protein repair mechanisms, and degradation of contaminants. Low energy consumption of cell adaptation is required to provide other physiological functions. Bacteria able to survive in toxic environment could help us to clean contaminated areas when they are used in bioremediation technologies. Slavomíra Murínová and Katarína Dercová Copyright © 2014 Slavomíra Murínová and Katarína Dercová. All rights reserved. Leptospirosis in Vellore: A Clinical and Serological Study Mon, 23 Jun 2014 06:32:37 +0000 Leptospirosis is a severe spirochetal zoonosis in the world. It is considered an occupational disease of persons engaged in agriculture, sewage works, forestry, and animal slaughtering. A study was conducted with an objective of assessing the seroprevalence of leptospirosis in and around Vellore district, Tamil Nadu. The study was based on the signs and symptoms of the patients reporting fever in Vellore Municipal Clinic (Urban Malarial Scheme). Blood samples were collected from 129 patients. Animal studies were conducted from 24 rodents captured form the market place of the Vellore municipality. In the ZET (Zonal Entomological Team, Vellore) Laboratory the sera were examined by macroscopic slide agglutination test (MSAT). In the MAST, totally 10 positive leptospiral cases from human beings and 10 positive leptospiral cases from rats (Rattus rattus and Rattus norvegicus) were found out. Then both positive cases of leptospiral vials were labeled, sealed, and sent to the Leptospirosis Research Laboratory, Madhavaram, Chennai, for further serovars examination. Among the various serovars identified autumnalis was more prevalent. Our findings showed that the age groups between 15 and 55 years showed more susceptibility. Particularly the adults were more infected. The majority of seropositive individuals in the cases had only subclinical infection. Rodents were abundant and contributed to enzootic and endemic prevalence of leptospirosis. G. Vimala, A. Mary Josephine Rani, and V. Raja Gopal Copyright © 2014 G. Vimala et al. All rights reserved. A Review on Antiulcer Activity of Few Indian Medicinal Plants Sun, 25 May 2014 06:51:32 +0000 Ulcer is a common gastrointestinal disorder which is seen among many people. It is basically an inflamed break in the skin or the mucus membrane lining the alimentary tract. Ulceration occurs when there is a disturbance of the normal equilibrium caused by either enhanced aggression or diminished mucosal resistance. It may be due to the regular usage of drugs, irregular food habits, stress, and so forth. Peptic ulcers are a broad term that includes ulcers of digestive tract in the stomach or the duodenum. The formation of peptic ulcers depends on the presence of acid and peptic activity in gastric juice plus a breakdown in mucosal defenses. A number of synthetic drugs are available to treat ulcers. But these drugs are expensive and are likely to produce more side effects when compared to herbal medicines. The literature revealed that many medicinal plants and polyherbal formulations are used for the treatment of ulcer by various ayurvedic doctors and traditional medicinal practitioners. The ideal aims of treatment of peptic ulcer disease are to relieve pain, heal the ulcer, and delay ulcer recurrence. In this review attempts have been made to know about some medicinal plants which may be used in ayurvedic as well as modern science for the treatment or prevention of peptic ulcer. G. Vimala and F. Gricilda Shoba Copyright © 2014 G. Vimala and F. Gricilda Shoba. All rights reserved. Antibacterial Activity of Murrayaquinone A and 6-Methoxy-3,7-dimethyl-2,3-dihydro-1H-carbazole-1,4(9H)-dione Tue, 20 May 2014 00:00:00 +0000 The antibacterial activity of Murrayaquinone A (10), a naturally occurring carbazoloquinone alkaloid, and 6-methoxy-3,7-dimethyl-2,3-dihydro-1H-carbazole-1,4(9H)-dione (11), a synthetic carbazoloquinone, both obtained during the development of the synthesis of Carbazomycin G, having unique quinone moiety, was studied against Gram-positive (Bacillus subtilis and Staphylococcus aureus) and Gram-negative (Escherichia coli and Pseudomonas sp.) bacteria. Compound 10 showed antibacterial activities against both of Escherichia coli and Staphylococcus aureus whereas compound 11 indicated the activity against Staphylococcus aureus only. Both compounds 10 and 11 exhibited minimum inhibitory concentration (MIC) of 50 μg mL−1 against Staphylococcus aureus. Biswanath Chakraborty, Suchandra Chakraborty, and Chandan Saha Copyright © 2014 Biswanath Chakraborty et al. All rights reserved. Diversity, Biocontrol, and Plant Growth Promoting Abilities of Xylem Residing Bacteria from Solanaceous Crops Mon, 19 May 2014 14:56:55 +0000 Eggplant (Solanum melongena L.) is one of the solanaceous crops of economic and cultural importance and is widely cultivated in the state of Goa, India. Eggplant cultivation is severely affected by bacterial wilt caused by Ralstonia solanacearum that colonizes the xylem tissue. In this study, 167 bacteria were isolated from the xylem of healthy eggplant, chilli, and Solanum torvum Sw. by vacuum infiltration and maceration. Amplified rDNA restriction analysis (ARDRA) grouped these xylem residing bacteria (XRB) into 38 haplotypes. Twenty-eight strains inhibited growth of R. solanacearum and produced volatile and diffusible antagonistic compounds and plant growth promoting substances in vitro. Antagonistic strains XB86, XB169, XB177, and XB200 recorded a biocontrol efficacy greater than 85% against BW and exhibited 12%–22 % increase in shoot length in eggplant in the greenhouse screening. 16S rRNA based identification revealed the presence of 23 different bacterial genera. XRB with high biocontrol and plant growth promoting activities were identified as strains of Staphylococcus sp., Bacillus sp., Streptomyces sp., Enterobacter sp., and Agrobacterium sp. This study is the first report on identity of bacteria from the xylem of solanaceous crops having traits useful in cultivation of eggplant. Gauri A. Achari and Raman Ramesh Copyright © 2014 Gauri A. Achari and Raman Ramesh. All rights reserved. Primary Otomycosis in the Indian Subcontinent: Predisposing Factors, Microbiology, and Classification Sun, 18 May 2014 11:07:57 +0000 Objective. To define otomycosis and determine the predisposing factors and microbiology in primary otomycosis. Study Design. Prospective study of two years and review of the literature. Setting. Academic Department of Otolaryngology in a coastal city in India. Patients. 150 immunocompetent individuals of whom 100 consecutive patients with a clinical diagnosis of otomycosis are considered as the study group and 50 consecutive patients with no otomycosis are considered as the control group. Results and Observations. Instillation of coconut oil (42%), use of topical antibiotic eardrops (20%), and compulsive cleaning of external ear with hard objects (32%) appeared to be the main predisposing factors in otomycosis. Aspergilli were the most common isolates (80%) followed by Penicillium (8%), Candida albicans (4%), Rhizopus (1%), and Chrysosporium (1%), the last being reported for the first time in otomycosis. Among aspergilli, A. niger complex (38%) was the most common followed by A. fumigatus complex (27%) and A. flavus complex (15%). Bacterial isolates associated with fungi in otomycosis were S. aureus, P. aeruginosa, and Proteus spp. In 42% of healthy external ears fungi were isolated. Conclusion. Aspergillus spp. were the most common fungi isolated, followed by Penicillium. Otomycotic ears are often associated with bacterial isolates when compared to normal ears. Fungi are also present in a significant number of healthy external auditory canals and their profiles match those in cases of otomycosis. The use of terms “primary” and “secondary” otomycosis is important to standardize reporting. Sampath Chandra Prasad, Subbannayya Kotigadde, Manisha Shekhar, Nikhil Dinaker Thada, Prashanth Prabhu, Tina D’ Souza, and Kishore Chandra Prasad Copyright © 2014 Sampath Chandra Prasad et al. All rights reserved. Molecular Typing of Methicillin Resistant Staphylococcus aureus Clinical Isolates on the Basis of Protein A and Coagulase Gene Polymorphisms Thu, 15 May 2014 11:09:21 +0000 Increased frequency of methicillin-resistant Staphylococcus aureus (MRSA) in hospitalized patients requires rapid and reliable characterization of isolates for control of MRSA spread in hospitals. This study evaluated polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) as a molecular typing technique for MRSA strains on the basis of protein A (spa) and coagulase (coa) gene polymorphisms to verify their ability in assessing the relatedness of isolates. Seventy-five MRSA isolates, from different ICUs of Alexandria University Main Hospital, were characterized using antibiotyping and PCR-RFLP analysis of coa and spa genes. Thirty-two antibiotypes were identified. coa gene PCR generated 3 types and 10 subtypes of band patterns. HaeIII restriction digestion of amplified coa gene products produced 5 major banding patterns and 12 subtypes. spa gene PCR products generated 4 major and 11 minor types, and their HaeII restriction digestion showed 5 major and 12 minor banding patterns. The combined coa and spa RFLP patterns generated 22 combined R types. Typing using coa PCR and PCR-RFLP had the same discriminatory index (DI) value (0.64), which was comparable to that of both spa PCR and PCR-RFLP techniques (0.68). The combined grouping increased the DI value to 0.836. The current study revealed that testing for multiple gene polymorphisms is more useful for local epidemiologic purposes. Nancy Younis Omar, Hala Abdel Salam Ali, Reem Abdel Hameed Harfoush, and Engy Hamdy El Khayat Copyright © 2014 Nancy Younis Omar et al. All rights reserved. Microbiological Quality of Salads Served along with Street Foods of Hyderabad, India Wed, 14 May 2014 13:27:05 +0000 A study has been done to analyse the microbiological quality of salads served along with street foods of Hyderabad. A total of 163 salad samples, 53 of carrot and 110 of onion samples, were collected from four different zones of Hyderabad. About 74% and 56% had Staphylococcus aureus in carrots and onions, respectively. Fifty-eight percent of carrots and forty-five percent of onions samples contained Salmonella, 68% of carrots and 24% of onions had Yersinia. HACCP study was carried out with 6 street food vendors to identify the source of Salmonella contamination in salads. Food handlers were found to be responsible for Salmonella contamination in salads. The present study revealed the potential hazards of street vended salad vegetables, considering the handling practice usually carried out by vendors. Ninety-eight percent of the vendors did not wash the vegetables before processing and serving while about 56.6% of the vendors did not peel the vegetables. Majority of street vendors’ nails were uncut. A significant difference () was observed in Yersinia spp. and Salmonella spp. in wet-dirty chopping board when compared to clean-dry chopping board. A significant difference () of Staphylococcus spp. was observed when the status of cleaning cloth was neat/untidy. Alekhya Sabbithi, R. Naveen Kumar, L. Kashinath, V. Bhaskar, and V. Sudershan Rao Copyright © 2014 Alekhya Sabbithi et al. All rights reserved. Study of a Natural Mutant SHV-Type β-Lactamase, SHV-104, from Klebsiella pneumoniae Tue, 13 May 2014 12:51:50 +0000 Klebsiella pneumoniae ML2011, a multiresistant isolate, was isolated from the Military Hospital of Tunis (Tunisia). The determination of the minimal inhibitory concentrations exhibited by K. pneumoniae ML2011 was performed by Etest. The crude extract of the isolates contains four different β-lactamases with pI 5.5, 7.3, 7.6, and 8.6. Only the β-lactamases with pI 7.3 and pI 8.6 were transferred by transformation and conjugation experiment. Molecular characterization of these genes was performed by PCR and sequencing. The chromosomal β-lactamases are TEM (pI 5.5) and SHV-1 (7.6). CTX-M-28 (pI 8.6) and the novel variant of SHV named SHV-104 (pI 7.3) were encoded by bla gene located on a 50 kb highly conjugative plasmid. The SHV-104 β-lactamase was produced in E. coli and purified. Its profile of activity was determined. Compared to SHV-1, SHV-104 contains one mutation, R202S. Their kinetic parameters were similar except for cefotaxime. The analysis of the predicted structure of SHV-104 indicated that the R202S mutation suppresses a salt bridge present in SHV-1. Therefore, the overall flexibility of the protein increased and might improve the hydrolysis of cefotaxime. We can conclude that the multiresistant phenotype of K. pneumoniae ML2011 strain is mainly linked to the production of CTX-M-28 since SHV-104 possesses a narrow spectrum of activity. Nahed Ben Achour, Omrane Belhadj, Moreno Galleni, Mohamed Ben Moussa, and Paola Sandra Mercuri Copyright © 2014 Nahed Ben Achour et al. All rights reserved. Prevalence of Clinical Periodontitis and Putative Periodontal Pathogens among South Indian Pregnant Women Mon, 12 May 2014 12:47:18 +0000 In view of recent understanding of the association of periodontal infections and adverse pregnancy outcomes, the present investigation was undertaken to study the periodontal infections among 390 asymptomatic pregnant women and to find an association of bacterial etiologies with the disease. Prevalence of gingivitis was 38% and clinical periodontitis was 10% among the study population. Subgingival plaque specimens were subjected to multiplex PCR targeting ten putative periodontopathogenic bacteria. Among the periodontitis group, high detection rates of Porphyromonas gingivalis (56%), Prevotella nigrescens (44%), Treponema denticola (32%), and Prevotella intermedius (24%) were noted along with significant association with the disease (). Chaitanya Tellapragada, Vandana Kalwaje Eshwara, Shashidhar Acharya, Parvati Bhat, Asha Kamath, Shashidhar Vishwanath, and Chiranjay Mukhopadhyay Copyright © 2014 Chaitanya Tellapragada et al. All rights reserved. Optimization of Amylase Production from B. amyloliquefaciens (MTCC 1270) Using Solid State Fermentation Sun, 11 May 2014 09:52:46 +0000 Demand for microbial amylase production persists because of its immense importance in wide spectrum industries. The present work has been initiated with a goal of optimization of solid state fermentation condition for amylase using agroindustrial waste and microbial strain like B. amyloliquefaciens (MTCC 1270). In an aim to improve the productivity of amylase, fermentation has been carried out in the presence of calcium (Ca+2), Nitrate (NO3−), and chloride ions (Cl−) as well as in the presence of D-inositol and mannitol. Amylase needs calcium ion for the preservation of its structure, activity and stability that proves beneficial also for amylase production using solid state fermentation. The inclusion of ions and sugars in the SSF media is promising which can be explained by the protection offered by them against thermal decay of amylase at various incubation periods at 37°C. Koel Saha, Sujan Maity, Sudeshna Roy, Koustav Pahan, Rishija Pathak, Susmita Majumdar, and Suvroma Gupta Copyright © 2014 Koel Saha et al. All rights reserved. Urinary Tract Infection among Antiretroviral Therapy Users and Nonusers in Jimma University Specialized Hospital, Jimma, Ethiopia Wed, 16 Apr 2014 13:57:41 +0000 Background. The introduction of antiretroviral therapy (ART) has dramatically reduced morbidity related with bacterial infection including urinary tract infection (UTI) among patients with HIV/AIDS. This study was carried out to determine the prevalence of UTI and identify common bacterial etiologies among HIV/AIDS patients of ART users and nonusers in Jimma University Specialized Hospital. Methods. A comparative cross sectional study was conducted from September to December 2012 on 367 ART users and 114 nonuser patients attending ART clinic. Sociodemographic characteristics, associated factors, and urine samples were collected; culture, biochemical tests, Gram stain, and drug sensitivity tests were done. Results. Of 467 examined urine samples, 56 (12%) had significant bacterial growth. Forty-six (12.5%) of the cases were ART users and 10 (10%) were nonusers. E. coli was the predominant isolate in both ART users (25 (54.3%)) and nonusers (6 (6%)). Majority of the bacterial isolates were from females. Most (>75%) of the isolates from both groups were resistant to ampicillin and trimethoprim-sulfamethoxazole but sensitive to norfloxacine, ceftriaxone, and chloramphenicol. Conclusion. There was no statistically significant difference regarding the prevalence of significant bacterial growth between ART users and nonusers. Therefore, it is recommended that UTI in both groups should be managed with drugs that show sensitivity. Serkadis Debalke, Waqtola Cheneke, Haimanot Tassew, and Mohammed Awol Copyright © 2014 Serkadis Debalke et al. All rights reserved. Transcriptional Response of Selenopolypeptide Genes and Selenocysteine Biosynthesis Machinery Genes in Escherichia coli during Selenite Reduction Tue, 15 Apr 2014 16:25:16 +0000 Bacteria can reduce toxic selenite into less toxic, elemental selenium (Se0), but the mechanism on how bacterial cells reduce selenite at molecular level is still not clear. We used Escherichia coli strain K12, a common bacterial strain, as a model to study its growth response to sodium selenite (Na2SeO3) treatment and then used quantitative real-time PCR (qRT-PCR) to quantify transcript levels of three E. coli selenopolypeptide genes and a set of machinery genes for selenocysteine (SeCys) biosynthesis and incorporation into polypeptides, whose involvements in the selenite reduction are largely unknown. We determined that 5 mM Na2SeO3 treatment inhibited growth by ∼50% while 0.001 to 0.01 mM treatments stimulated cell growth by ∼30%. Under 50% inhibitory or 30% stimulatory Na2SeO3 concentration, selenopolypeptide genes (fdnG, fdoG, and fdhF) whose products require SeCys but not SeCys biosynthesis machinery genes were found to be induced ≥2-fold. In addition, one sulfur (S) metabolic gene iscS and two previously reported selenite-responsive genes sodA and gutS were also induced ≥2-fold under 50% inhibitory concentration. Our findings provide insight about the detoxification of selenite in E. coli via induction of these genes involved in the selenite reduction process. Antonia Y. Tetteh, Katherine H. Sun, Chiu-Yueh Hung, Farooqahmed S. Kittur, Gordon C. Ibeanu, Daniel Williams, and Jiahua Xie Copyright © 2014 Antonia Y. Tetteh et al. All rights reserved. Genotyping of ESBL Producing Uropathogenic Escherichia coli in West of Iran Tue, 15 Apr 2014 08:22:59 +0000 Background and Objective. Urinary tract infection (UTI) is one of the most common bacterial infections in the world. Molecular fingerprinting of UTI isolates such as pulsed-Field Gel Electrophoresis using for Clonal distribution and determine of predominant type. The aim of the study was to determine genotyping of ESBL producing UPECs. Material and Methods. 200 UPEC isolates from outpatients with UTI were obtained. Antimicrobial susceptibility and interpretation were performed by disk diffusion. Virulence factors for UPECs were screened by using PCR. UPECs were analyzed by Pulsed-Field Gel Electrophoresis and images analyzed by Phoretix1DPro software. Results. A total of 200 isolates of UPECs, 24.5% () of isolates, were positive for ESBL production. Resistance ranged from 0% for amikacin and imipenem to over 93.9% for carbenicillin and ampicillin. Frequencies of haemagglutination, haemolysin, and hydrophobicity were 51%, 18.3%, and 14.28%, respectively. A total of 10 different genotypes were obtained, which include nine common clones and one single clone. Conclusion. We confirmed the prevalence of virulence phenotyping especially Haemagglutination among UPEC strains and that it can also contribute to virulence in these strains. Large diversity in genotypes was observed in the isolates that could be indicative of different sources of infection in community acquired. Parviz Mohajeri, Gita Darfarin, and Abbas Farahani Copyright © 2014 Parviz Mohajeri et al. All rights reserved. Removal of Total Coliforms, Thermotolerant Coliforms, and Helminth Eggs in Swine Production Wastewater Treated in Anaerobic and Aerobic Reactors Wed, 09 Apr 2014 08:34:18 +0000 The present work evaluated the performance of two treatment systems in reducing indicators of biological contamination in swine production wastewater. System I consisted of two upflow anaerobic sludge blanket (UASB) reactors, with 510 and 209 L in volume, being serially arranged. System II consisted of a UASB reactor, anaerobic filter, trickling filter, and decanter, being also organized in series, with volumes of 300, 190, 250, and 150 L, respectively. Hydraulic retention times (HRT) applied in the first UASB reactors were 40, 30, 20, and 11 h in systems I and II. The average removal efficiencies of total and thermotolerant coliforms in system I were 92.92% to 99.50% and 94.29% to 99.56%, respectively, and increased in system II to 99.45% to 99.91% and 99.52% to 99.93%, respectively. Average removal rates of helminth eggs in system I were 96.44% to 99.11%, reaching 100% as in system II. In reactor sludge, the counts of total and thermotolerant coliforms ranged between 105 and 109 MPN (100 mL)−1, while helminth eggs ranged from 0.86 to 9.27 eggs g−1 TS. Silvia Helena Zacarias Sylvestre, Estevam Guilherme Lux Hoppe, and Roberto Alves de Oliveira Copyright © 2014 Silvia Helena Zacarias Sylvestre et al. All rights reserved. Virulence Factors Contributing to Pathogenicity of Candida tropicalis and Its Antifungal Susceptibility Profile Wed, 02 Apr 2014 09:52:11 +0000 The incidence of invasive candidiasis has increased over the past few decades. Although Candida albicans remains by far the most common species encountered, in recent years shift towards non-albicans Candida species like Candida tropicalis is noted. Here in this study we determined the virulence factors and antifungal susceptibility profile of 125 C. tropicalis isolated from various clinical specimens. Biofilm formation was seen in 53 (42.4%) isolates. Coagulase production was noted in 18 (14.4%) isolates. Phospholipase enzyme was the major virulent factor produced by C. tropicalis isolates. A total of 39 biofilm forming isolates showed phospholipase activity. Proteinase activity was demonstrated by 65 (52%) isolates. A total of 38 (30.4%) isolates showed haemolytic activity. Maximum isolates demonstrated resistance to fluconazole. Fluconazole resistance was more common in C. tropicalis isolated from blood cultures. Antifungal resistance was more in isolates possessing the ability to produce phospholipase and biofilm. C. tropicalis exhibit a great degree of variation not only in their pathogenicity but also in their antifungal susceptibility profile. The identification of virulence attributes specific for each species and their correlation with each other will aid in the understanding of the pathogenesis of infection. Sachin C. Deorukhkar, Santosh Saini, and Stephen Mathew Copyright © 2014 Sachin C. Deorukhkar et al. All rights reserved. Lack of Outer Membrane Protein A Enhances the Release of Outer Membrane Vesicles and Survival of Vibrio cholerae and Suppresses Viability of Acanthamoeba castellanii Tue, 01 Apr 2014 09:13:57 +0000 Vibrio cholerae, the causative agent of the diarrhoeal disease cholera, survives in aquatic environments. The bacterium has developed a survival strategy to grow and survive inside Acanthamoeba castellanii. It has been shown that V. cholerae expresses outer membrane proteins as virulence factors playing a role in the adherence to interacted host cells. This study examined the role of outer membrane protein A (OmpA) and outer membrane vesicles (OMVs) in survival of V. cholerae alone and during its interaction with A. castellanii. The results showed that an OmpA mutant of V. cholerae survived longer than wild-type V. cholerae when cultivated alone. Cocultivation with A. castellanii enhanced the survival of both bacterial strains and OmpA protein exhibited no effect on attachment, engulfment, and survival inside the amoebae. However, cocultivation of the OmpA mutant of V. cholerae decreased the viability of A. castellanii and this bacterial strain released more OMVs than wild-type V. cholerae. Surprisingly, treatment of amoeba cells with OMVs isolated from the OmpA mutant significantly decreased viable counts of the amoeba cells. In conclusion, the results might highlight a regulating rule for OmpA in survival of V. cholerae and OMVs as a potent virulence factor for this bacterium towards eukaryotes in the environment. Soni Priya Valeru, Salah Shanan, Haifa Alossimi, Amir Saeed, Gunnar Sandström, and Hadi Abd Copyright © 2014 Soni Priya Valeru et al. All rights reserved. Antibiofilm Activity, Compound Characterization, and Acute Toxicity of Extract from a Novel Bacterial Species of Paenibacillus Mon, 24 Mar 2014 07:10:30 +0000 The effectiveness of many antimicrobial agents is currently decreasing; therefore, it is important to search for alternative therapeutics. Our study was carried out to assess the in vitro antibiofilm activity using microtiter plate assay, to characterize the bioactive compounds using Ultra Performance Liquid Chromatography-Diode Array Detection and Liquid Chromatography-Mass Spectrometry and to test the oral acute toxicity on Sprague Dawley rats of extract derived from a novel bacterial species of Paenibacillus strain 139SI. Our results indicate that the crude extract and its three identified compounds exhibit strong antibiofilm activity against a broad range of clinically important pathogens. Three potential compounds were identified including an amino acid antibiotic C8H20N3O4P (MW 253.237), phospholipase A2 inhibitor C21H36O5 (MW 368.512), and an antibacterial agent C14H11N3O2 (MW 253.260). The acute toxicity test indicates that the mortality rate among all rats was low and that the biochemical parameters, hematological profile, and histopathology examination of liver and kidneys showed no significant differences between experimental groups . Overall, our findings suggest that the extract and its purified compounds derived from novel Paenibacillus sp. are nontoxic exhibiting strong antibiofilm activity against Gram-positive and Gram-negative pathogens that can be useful towards new therapeutic management of biofilm-associated infections. Saad Musbah Alasil, Rahmat Omar, Salmah Ismail, and Mohd Yasim Yusof Copyright © 2014 Saad Musbah Alasil et al. All rights reserved. Detection of Integrase Gene in E. coli Isolated from Pigs at Different Stages of Production System Mon, 10 Mar 2014 12:12:49 +0000 Integrons are one of the genetic elements involved in the acquisition of antibiotic resistance. The aim of the present research is to investigate the presence of integrons in commensal Escherichia coli (E. coli) strains, isolated from pigs at different stages of production system and from the environment in an Argentinian farm. Five sows postpartum and five randomly chosen piglets from each litter were sampled by rectal swabs. They were sampled again at day 21 and at day 70. Environmental samples from the farm were also obtained. E. coli containing any integron class or combination of both integrons was detected by polymerase chain reaction in 100% of sows and in piglets at different stages of production: farrowing pen stage 68.1%;, weaning 60%, and growing/finishing 85.8%, showing an increase along the production system. From environmental samples 78.4% of E. coli containing any integron class was detected. We conclude that animals and farm environment can act as reservoirs for potential spread of resistant bacteria by means of mobile genetic elements as integrons, which has a major impact on production of food animals and that can reach man through the food chain, constituting a problem for public health. Eulalia de la Torre, Rocío Colello, Nora Lía Padola, Analía Etcheverría, Edgardo Rodríguez, Fabián Amanto, María Ofelia Tapia, and Alejandro Luis Soraci Copyright © 2014 Eulalia de la Torre et al. All rights reserved. Escherichia coli Strains Isolated from the Uteri Horn, Mouth, and Rectum of Bitches Suffering from Pyometra: Virulence Factors, Antimicrobial Susceptibilities, and Clonal Relationships among Strains Sun, 09 Mar 2014 12:02:23 +0000 Pyometra is recognized as one of the main causes of disease and death in the bitch, and Escherichia coli is the major pathogen associated with this disease. In this study, 70 E. coli isolates from the uteri horn, mouth, and rectum of bitches suffering from the disease and 43 E. coli isolates from the rectum of clinically healthy bitches were examined for the presence of uropathogenic virulence genes and susceptibility to antimicrobial drugs. DNA profiles of isolates from uteri horn and mouth in bitches with pyometra were compared by REP, ERIC, and BOX-PCR. Virulence gene frequencies detected in isolates from canine pyometra were as follows: 95.7% fim, 27.1% iss, 25.7% hly, 18.5% iuc, and 17.1% usp. Predominant resistance was determined for cephalothin, ampicillin, and nalidixic acid among the isolates from all sites examined. Multidrug resistance was found on ∼50% pyometra isolates. Using the genotypic methods some isolates from uteri, pus, and saliva of the same bitch proved to have identical DNA profiles which is a reason for concern due to the close relationship between household pets and humans. Juliana M. A. Agostinho, Andressa de Souza, Ruben P. Schocken-Iturrino, Lívia G. Beraldo, Clarissa A. Borges, Fernando A. Ávila, and José M. Marin Copyright © 2014 Juliana M. A. Agostinho et al. All rights reserved. Erratum to “Design and Optimization of a Process for Sugarcane Molasses Fermentation by Saccharomyces cerevisiae Using Response Surface Methodology” Tue, 25 Feb 2014 09:10:17 +0000 Nour Sh. El-Gendy, Hekmat R. Madian, and Salem S. Abu Amr Copyright © 2014 Nour Sh. El-Gendy et al. All rights reserved. Type IV Secretion System Is Not Involved in Infection Process in Citrus Sun, 23 Feb 2014 07:49:13 +0000 The type IV secretion system (T4SS) is used by Gram-negative bacteria to translocate protein and DNA substrates across the cell envelope and into target cells. Xanthomonas citri subsp. citri contains two copies of the T4SS, one in the chromosome and the other is plasmid-encoded. To understand the conditions that induce expression of the T4SS in Xcc, we analyzed, in vitro and in planta, the expression of 18 ORFs from the T4SS and 7 hypothetical flanking genes by RT-qPCR. As a positive control, we also evaluated the expression of 29 ORFs from the type III secretion system (T3SS), since these genes are known to be expressed during plant infection condition, but not necessarily in standard culture medium. From the 29 T3SS genes analyzed by qPCR, only hrpA was downregulated at 72 h after inoculation. All genes associated with the T4SS were downregulated on Citrus leaves 72 h after inoculation. Our results showed that unlike the T3SS, the T4SS is not induced during the infection process. Tiago Rinaldi Jacob, Marcelo Luiz de Laia, Leandro Marcio Moreira, Janaína Fernandes Gonçalves, Flavia Maria de Souza Carvalho, Maria Inês Tiraboschi Ferro, and Jesus Aparecido Ferro Copyright © 2014 Tiago Rinaldi Jacob et al. All rights reserved. Antibacterial Activities and In Vitro Anti-Inflammatory (Membrane Stability) Properties of Methanolic Extracts of Gardenia coronaria Leaves Wed, 19 Feb 2014 00:00:00 +0000 This work is carried out with Gardenia coronaria leaves that belong to the family Rubiaceae, which is a small-to-medium-sized but tall, deciduous tree, 7.6–9 m high on an average. Leaves are used for the treatment of rheumatic pain and bronchitis. The leaf of the plant consists of coronalolide, coronalolic acid, coronalolide methyl ester, ethyl coronalolate acetate triterpenes (secocycloartanes), and so forth. Methanol extract from the leaves of Gardenia coronaria was completely screened for membrane stability and antibacterial activity. The lower concentrations of Methanolic leaf extract of Gardenia coronaria gave good antimicrobial and anti-inflammatory activity, but higher concentrations gave relatively more projecting antibacterial activity in vitro as compared with Kanamycin. The crude drug’s anti-inflammatory effects were compared with those of Aspirin as positive control. The Methanolic extracts of Gardenia coronaria leaves possessed a broad spectrum antibacterial activity against a variety of both Gram-negative and Gram-positive organisms like Streptococcus agalactiae, Escherichia coli, Pseudomonas aeruginosa, Bacillus cereus, Shigella sonnei, Shigella boydii, and Proteus mirabilis, with a zone of inhibition from 10 to 16 mm. The extract also showed good membrane stability to be considered as having significant anti-inflammatory action. Amin Chowdhury, Shofiul Azam, Mohammed Abdullah Jainul, Kazi Omar Faruq, and Atiqul Islam Copyright © 2014 Amin Chowdhury et al. All rights reserved. False-Negative Rate of Gram-Stain Microscopy for Diagnosis of Septic Arthritis: Suggestions for Improvement Thu, 13 Feb 2014 08:17:14 +0000 We quantify the false-negative diagnostic rate of septic arthritis using Gram-stain microscopy of synovial fluid and compare this to values reported in the peer-reviewed literature. We propose a method of improving the diagnostic value of Gram-stain microscopy using Lithium Heparin containers that prevent synovial fluid coagulation. Retrospective study of the Manchester Royal Infirmary microbiology database of patients undergoing synovial fluid Gram-stain and culture between December 2003 and March 2012 was undertaken. The initial cohort of 1896 synovial fluid analyses for suspected septic arthritis was reduced to 143 after exclusion criteria were applied. Analysis of our Gram-stain microscopy yielded 111 false-negative results from a cohort size of 143 positive synovial fluid cultures, giving a false-negative rate of 78%. We report a false-negative rate of Gram-stain microscopy for septic arthritis of 78%. Clinicians should therefore avoid the investigation until a statistically significant data set confirms its efficacy. The investigation's value could be improved by using Lithium Heparin containers to collect homogenous synovial fluid samples. Ongoing research aims to establish how much this could reduce the false-negative rate. Paul Stirling, Radwane Faroug, Suheil Amanat, Abdulkhaled Ahmed, Malcolm Armstrong, Pankaj Sharma, and Ahmed Qamruddin Copyright © 2014 Paul Stirling et al. All rights reserved. Molecular Analysis of Human Metapneumovirus Detected in Patients with Lower Respiratory Tract Infection in Upper Egypt Thu, 30 Jan 2014 11:27:22 +0000 Introduction. Since 2001, when Human metapneumovirus (HMPV) was isolated in the Netherlands, the virus has been detected in several continents. Although reports have confirmed the prevalence of HMPV worldwide, data from Egypt remain limited. HMPV plays an important role in respiratory tract infections in individuals of all ages particularly in children. This study was aimed at estimating the prevalence of HMPV in patients with community-acquired lower respiratory infection in Upper Egypt and characterizing the circulating Egyptian HMPV strains for the first time. Materials and Methods. From 2005 to 2008, respiratory samples from 520 patients were analyzed for the presence of HMPV by real-time RT-PCR. Molecular and phylogenetic analyses were performed on partial fusion gene sequences of HMPV-positive patients. Results. HMPV-positive patients were detected in 2007-2008. The overall infection rate was 4%, while 57% of the patients were children. Sequence analysis demonstrated circulation of subgroup B viruses with predominance of lineage B2. Nucleotide sequence identity within lineage B1 was 98.8%–99.7% and higher than that in lineage B2 (94.3%–100%). Three new amino acid substitutions (T223N, R229K, and D280N) of lineage B2 were observed. Conclusion. HMPV is a major viral pathogen in the Egyptian population especially in children. During 2007-2008, predominantly HMPV B2 circulated in Upper Egypt. Mona S. Embarek Mohamed, Janine Reiche, Sonja Jacobsen, Amany G. Thabit, Mohamed S. Badary, Wolfram Brune, Brunhilde Schweiger, and Ahmed H. Osmann Copyright © 2014 Mona S. Embarek Mohamed et al. All rights reserved. Simultaneous Production of Biosurfactants and Bacteriocins by Probiotic Lactobacillus casei MRTL3 Wed, 29 Jan 2014 00:00:00 +0000 Lactic acid bacteria (LAB) are ubiquitous and well-known commensal bacteria in the human and animal microflora. LAB are extensively studied and used in a variety of industrial and food fermentations. They are widely used for humans and animals as adjuvants, probiotic formulation, and dietary supplements and in other food fermentation applications. In the present investigation, LAB were isolated from raw milk samples collected from local dairy farms of Haryana, India. Further, the isolates were screened for simultaneous production of biosurfactants and bacteriocins. Biosurfactant produced was found to be a mixture of lipid and sugar similar to glycolipids. The bacteriocin obtained was found to be heat stable (5 min at 100°C). Further, DNA of the strain was extracted and amplified by the 16S rRNA sequencing using universal primers. The isolate Lactobacillus casei MRTL3 was found to be a potent biosurfactant and bacteriocin producer. It seems to have huge potential for food industry as a biopreservative and/or food ingredient. Deepansh Sharma and Baljeet Singh Saharan Copyright © 2014 Deepansh Sharma and Baljeet Singh Saharan. All rights reserved.