API 20E System Research Articles

The use of Omp W Gene in Detection of Vibrio cholerae Isolated from Diarrhea Cases of Children in AL-Diwaniya Province

A total of 221 stool samples were collected from children suffering from watery diarrhea, less than 15 years old of both genders whom admitted to the Maternity and Children Teaching Hospital in Al- Diwaniya Province, In addition to that, 27 water samples were also collected from three different loci of ALDiwaniya River, at the period of October 2008 to June 2009,in order to evaluate the routine laboratory diagnostic procedures in the diagnosis the multi-serogroups or serotype of Vibrio cholerae strains and compared them with molecular technique as Polymerase chain reaction (PCR).Vibrio cholerae has been isolated and identified by using culturing method in addition to biochemical tests, API 20E diagnostic kit. Serotyping by using polyvalent Vibrio cholerae, O1antisera, and monovalent Ogawa and Inaba revealed that the most of clinical V. cholerae were of serogroup O1, while all the V. cholerae isolated from surface water of AL- Diwanyia river was Non-O1 serogroup. PCR technique was used to detect ompW gene encoding to outer membrane protein of V. cholerae. Based on the PCR results, the rate of Vibrio cholerae isolation from stool samples was 5.9%. PCR results showed that there were high specificity (100%, 100%, 97% and 86%) in detection of Vibrio cholerae strains versus each of cultural, biochemical, API 20E system and serological tests, respectively.

Reciprocal Intermolecular antigenic competition between E. coli and P. aeruginosa

It has been observed that in a dimicrobic mucosal urinary tract infection . The specific antibody titre of E. coli was higher than that of P. aeruginosa. The interpretation was based on antigenic competition . Hence a case of UTI with combined gram negative infection were chosen and diagnosed as E. coli and P. aeruginosa (Identified by classical tests and API 20E system) . Balanced and unbalanced heat killed E. coli and P. aeruginosa antigens in different combination proportions were made and used for specific immune priming of rabbits. Monotypic heat killed antigens were made and used for priming rabbits separately standard tube agglutination and ELISA IL-4 were used to match humoral and cellular arms respectively in the state of secondary immune responses. The specific antibody mean titres in combinations was reduced as compared to mean titres of monotypic sera which is an indication for antigenic competition . It was reciprocal intermolecular type (T cell dependent) . IL-4 determinations were increased as compared monotypic and control in the sera react against unbalanced combinations. Such increase indicate its role in antigenic competition . Thus , the E. coli-P. aeruginosa antigenic competition is a reciprocal , humoral and cellular and intermolecular type. The role of IL-4 in antigenic competition is being reported for the first time .

Radiation Response (Low Dose Gamma Radiation) of Members of the Enterobacteriaceae in Dry Fruiting Bodies of Pleurotus ostreatus and Their Identification Using API 20E System

The effectiveness of using low dose gamma radiation (1, 2, 3, 4 and 5 kiloGray) was tested on dry fruit bodies of Pleurotus ostreatus stored in polythene and polypropylene and stored at 28-30 oC up to 12 months for identification of members of Enterobacteriaceae with API 20E test kit (Biomerieux ®, France). Encountered microorganisms of the Control samples (0 kGy) of both packages recorded included Klebsiella pneumoniae, Citrobacter freundii, Pseudomonas aeroginosa Proteus mirabilis, Serratia marscesens and Enterobacter spp. There was an observed decrease in bacteria species with increasing dose. However, Klebsiella pneumonia persisted after the application 5 kGy dose in both packaging materials. Health and immuno compromised persons stand a greater health risk after interaction with some of these bacteria spp. Case in point, infection of patients with cystic fibrosis by Pseudomonas aeruginosa is known to be lethal over a long period of time. Information on Enterobacteria in the food industry is essential since they are potential food poisoning agents and thus their control is imperative.

Frequency of Metallo-β-Lactamases Producing Klebsiella pnumoniae in Burn Wound Isolates

From the period 1 /3 /2016 to 30 / 8 / 2016, 210 swabs were collected from the burn patients hospitalized in different hospitals in Baghdad City: Al-Karama Teaching Hospital, Special Burn Hospital, Central Teaching Laboratories, Child protection Teaching Hospital, Imam Ali Hospital. Forty two isolates (37.5 %) showed a growth of Klebsiella pneumonia, and the remaining isolates (62.5%) were belonged to other bacteria. Identification of all isolates were carried out depending on macroscopic, microscopic characterizations, conventional biochemical tests and Api 20E system. Metallo -β lactamase (MBL) enzymes were screen by three different phenotypic methods (DDST, CMDT and MHT) in which 10µg meropenem antibiotic disk has selected to all phenotypic methods. Susceptibility test done with antibiotic disks of cefotaxime, ceftazidime, aztreonam, imipenem, meropenem, gentamicin, amikacin, ciprofloxacin, tetracycline and trimethoprim-sulfamethoxazole. The percentage of resistance isolates were presented as, Tetracycline (95.23%), Cefotaxime (85.71%), Trimethoprim- Sulfamethoxazol (83.33%), Aztreonam (71.42%), Ceftazidime (69.04%), Ciprofloxacin (59.52%), Gentamycin (26.19%), Imipenem (21.42%), Meropenem and Amikacin (19.04 %).

HIGH VAGINAL SWABS;

Normal vaginal flora contains a wide range of microorganisms. Hydrogen peroxideproduced by Lactobacillus strains plays a vital role in maintaining the microenvironment of thevagina and in the inhibition of overgrowth of potentially pathogenic bacteria. Bacterial vaginosisBV is the main reason of vaginal discharge. Many gram positive and gram negative rods i.e.E.coli, Klebsiella, Proteus, Acinetobacter and Pseudomonas spp. are major contributors inbacterial vaginosis. Aim: The present study was conducted to elucidate the frequency of variousgram-negative rods in high vaginal swabs and sensitivity pattern of bacteria to antibiotics thatare currently used. Study Design: Cross sectional study. Setting: Department of Obstetricsand Gynecology of Benazir Bhutto Hospital Rawalpindi, a tertiary health care center for thepeople of Rawalpindi. Period: January 2015 to May 2016. Material and Methods: A total of220 High vaginal swabs (HVS) were collected both from indoor and outdoor patients presentingwith symptoms of vaginal discharge aged between 20 to 65 years. Swabs were inoculated onblood, Chocolate and MacConkey’s agar. After overnight incubation plates were examined forgrowth, colonial morphology, final confirmation was done on the basis of biochemical testingand API 20-E system (BioMerieux, France) up to species level. Antibiotic sensitivity testing wasdone by (modified Kirby-Bauer’s) disc diffusion method using amikacin, ampicillin, amoxicillinclavulanic acid, imipenem, ceftazidime, tigecycline, ciprofloxacin, sulzone and cefixime. Afterovernight incubation plates were examined to read the susceptibility zone. Results: Out of 220HVS samples, 100 samples showed bacterial growth and confirmed as Gram negative bacilli.Age wise distribution of infection showed highest rates b/w age 20-30 was 36% followed by 31-40 (23%), 41-50 (25%) and 11% above 50 years of age. Bacteria isolated from HVS were E.coli(53%), Klebsiella (22%), Pseudomonas (12%), citrobacter (6%), Proteus (5%) and Acinetobacter(2%) respectively. Highly sensitive antibiotics against bacteria were imipenem (96%), sulzone(90%) and Ciprofloxacin (88%), whereas least affective antibiotics against gram negative rodswere penicillins (ampicillin, amoxicillin-clavulanic acid), amikacin due to indiscriminate use ofantibiotics. Conclusion: High prevalence of gynecological infections demands that the patientswho have vaginosis must be investigated regularly and carefully through culture and identificationof causative bacteria. Emergence of antibiotic resistance must be controlled in order to avoidimproper use, frequent abuse, insufficient dosages, trouble-free availability of antibiotics andtreatment schedule must be designed subsequent to proper laboratory investigations.

Physicochemical Factors Affected the Partial Purified Lipase Activity of Acinetobacter baumannii "local isolates"

Microbial lipases today occupy a place of prominence among biocatalysts owing to their ability to catalyze awide variety of reactions in aqueous and non- aqueous media, A.baumannii were isolated from different clinical specimens from hospitalized patients from Baghdad hospitals and were detected by biochemical tests and API20E system. The percentage of isolation was (16.6%), A. baumannii is an increasingly multidrug – resistant (MDR), it showed high level of resistant to Ceftriaxon, Colistin, Piperacillin, Co-trimoxazol, Tertracycline, Carbenicillin, Amoxicillin, Penicillin G, Gentamicin and Ceftazidim , wherease the isolates were highly sensitive to Imipenem, Ciprofloxacin, Meropenem, Amikacin, and Cefotaxime.
 The isolated strians of A.baumannii were screened for the production of lipase by using Rhodamine B agar media. one of the isolated strians exhibited a greater clear zone than the others, indicated higher lipase activity.
 The lipase in present study was partially purified by ammonium sulphate at the concenterations of 30% and 80%, the 30% ammonium sulphate showed higher activiy of the enzyme than 80% in which the enzyme activity was slightly decreased indicating that the better purification was at the 30% concenteration of ammonium sulphate.Various physicochemical parameters such as pH, temperature and metal ions were studied in order to determine the optimum conditions for lipase production.
 The production of lipase by A. baumannii was optimum at 35°C, pH 7.0 and was enhanced by the Ca++ and Na+ wherease inhibited by Zn++ ions.
 Key words : Acinetobacter baumannii, Lipase enzyme, Partial purification.

Isolation and Identification of protease producing Aeromonas hydrophila

A total of 89 samples were collected from pools of fish farms and gills pieces of fresh fishes from different locations in Baghdad governorate. From these samples, a total of 91 bacterial isolates were obtained and identified as suspected Aeromonas spp. according to the cultural and microscopical characteristics. After subjection to the biochemical tests, seven isolates were identified as Aeromonas hydrophila. Results of identification were confirmed by using Api 20E system. Ability of these isolates for protease production was screened. Results showed that all the seven isolates were protease producers; among them the A. hydrophila A4 obtained from gills pieces of fresh fishes was the most efficient in protease production. Enzyme specific activity of protease of the crude filtrate of this isolate was 31.01U/mg protein.

Intense pre-admission carriage and further acquisition of ESBL-producing Enterobacteriaceae among patients and their caregivers in a tertiary hospital in Rwanda.

To assess the presence and risk factors of intestinal carriage of extended-spectrum beta-lactamase-producing Enterobacteriaceae (ESBL-PE) among patients admitted to the University Teaching Hospital of Butare and among their attending caregivers, and to analyse the acquisition of ESBL-PE carriage during hospital stay and associated factors. We screened 392 patients and their attending caregivers at admission and discharge for ESBL-PE carriage. Bacterial species were determined using the API-20E system, and antimicrobial susceptibility testing was performed by agar disc diffusion. Data on socio-economic status, diet, behaviour, household assets, livestock and hospital procedures were collected. At admission, 50% of the patients showed intestinal ESBL-PE carriage (Escherichia coli, 51%; Klebsiella pneumoniae, 39%; Enterobacter cloacae, 19%) as did 37% of their caregivers. Co-resistance was common but no carbapenem resistance was detected. At discharge, the proportion of ESBL-PE-colonised patients increased to 65% (caregivers, 47%) with almost complete carriage in paediatric patients (93%). The acquisition rate among initially non-colonised patients was 55% (or, 71/1000 patient days). Independent predictors of admission carriage included a colonised caregiver, prior antibiotic intake, egg consumption and neglecting to boil drinking water, whereas being a paediatric patient, undergoing surgery and male gender predicted acquisition during hospitalisation. Abundant admission carriage of ESBL-PE and a high acquisition rate in a Rwandan university hospital point to potential intrahospital transmission and community dissemination. Caregivers are an additional source of possible spread. Risk factors of colonisation such as diet and water source need to be tackled to prevent the further emergence and spread of ESBL-PE.

Histopathological Changes in Diseased Catfish (Clarias gariepinus) and Treated by Ciprofloxacin and Clove Extract

Bacteria pathogens may cause a serious loss in aquaculture and also health hazards to humans. On the basis of biochemical characteristics and API-20E system, the highest prevailing isolate was identified as Aeromonas sobria. Infection with Aeromonas sobria in African catfish (Clarias gariepinus) induced focal hemorrhage at the end of gill operculum, generalized edema and ulcers on skin surface. Internal signs of infection includes hyperemia of the hepatic artey, coagulative necrosis in the liver and hydropic degeneration of renal tubules according to the histopathological examination. In-vivo the combination of ciprofloxacin and clove extract was the choice as an alternative bacterial therapeutic agents in infected Clarias gariepinus with Aeromonas sobria. Moreover histopathological changes have is the base for the evaluation of fish health.

Effect of Antibiotic Combinations on the Sensitivity of Carbapenem Resistant Acinetobacter baumannii Strains

Acinetobacter baumannii is one of the predominant pathogens involved in hospital acquired infections worldwide. Despite intensive surveillance and preventive measures, these infections are major cause of high mortality rates. In this study a total of 375 A. baumannii isolates were isolated from a variety of clinical collected from 5 different hospitals in Great Cairo. These isolates were identified as A. baumannii biochemically, by API20E system and genetically by detection of 16S rRNA gene. All isolates showed positive results and confirmed for the presence of the suspected gene by PCR. The antibiotic susceptibility patterns for the isolated A. baumannii were also evaluated. The detection of the presence of OXA-type (OXA 23, 24, 51 and 58) carbapenemases-encoding genes which are responsible for carbapenem resistance was also performed. I observed the presence of OXA 23 gene in 84% of the isolates. While 35.2% were positive to OXA 24 gene and 87.2% were positive to OXA 51 gene. No isolates showed positive results for the presence of OXA 51 gene. They were also evaluated for the effectiveness of drug combination of carbapenems and colistin against 30 isolates. Regarding imipenem and colistin combination, 13.3% of the strains showed synergy, while 86.7% showed additive results. For the combination of meropenem and colistin, 66.7% of the strains showed synergy, while 33.3% showed additive results. The antibacterial effect of both combinations on A. baumannii showed commonly synergistic or additive results. To a significant extent meropenem and colistin showed a superior synergy compared to imipenem and colistin.

Detection of Efflux Pumps Genes in Clinical Isolates of Acinetobacter baumannii

One of the most interesting features of Acinetobacter baumannii is the ease by which it can acquire resistance to various antibiotics. Mutations, new gene acquisitions, up-regulating expression of existing genes (should those be genes of drug-destroying enzymes or efflux pumps), or losing them, like in the case of porins, all may contribute to the broad spectrum resistance of these strains.This study was directed to decide the event of efflux pumpsgenes harboring A. baumannii isolates recovered from Babylon hospitals. Isolates were recognized by biochemical tests and more affirmed utilizing API 20E system. Antibioticssusceptibility was measured by utilizing disks diffusion test. Ten (0.76%) A. baumannii isolates were recovered from clinical specimens. One (10%) isolate was observed to be imipenem and meropenem resistant (MIC > 512 µg/ml). There are indicative appearance of efflux pump genes ; (50%) for Ade-A and Ade-B each other , (40%) for Ade-C, (80%) for all of Ade-R and Ade-S , (100%) for all of Ade-I and Ade-J , and (50%) for Ade-Y. All isolates of A. baumannii appear as MDR , while only one appear to be as PDR.

Antimicrobial effect of the Tunisian Nana variety Punica granatum L. extracts against Salmonella enterica (serovars Kentucky and Enteritidis) isolated from chicken meat and phenolic composition of its peel extract

Punica granatum L. is widely recognized for its potency against a broad spectrum of bacterial pathogens. The purpose of this study was to explore the inhibitory and the bactericidal activities of Punica granatum against Salmonella strains. The effect of extracts obtained from different parts (peels, seeds, juice and flowers) of pomegranate and using different solvents against Salmonella enterica serovars Kentucky and Enteritidis isolated from chicken meat was thus investigated. Salmonella strains were identified with the standard API-20E system and confirmed by real time PCR.The obtained results showed that the highest antibacterial activity against Salmonella strains was observed with the peels ethanolic extract giving MIC values ranging from 10.75 to 12.5mg/mL. The ethanolic extract of P. granatum Nana peels at 0.8 and 1.6mg/g significantly inhibited the growth of Salmonella Kentucky in chicken meat stored at 4°C.The phenolic composition of the ethanolic peel extract was explored by HPLC coupled to both DAD and ESI/TOF-MS detections. The obtained results allowed the detection of 21 phytochemical compounds among which various phenolic compounds have been identified on the basis of their UV and MS spectra as well as with literature data. Among the detected compounds, anthocyanins, ellagitannins, ellagic acid derivatives and flavanols were further characterized through MS-MS analysis.Our results showed thus that the Tunisian variety Nana pomegranate constitutes a good source of bioactive compounds with potent antimicrobial activity on the growth of Salmonella strains suggesting that the studied pomegranate cultivar could be a natural remedy to minimize the emergence of Salmonella enterica strains which is often involved in food borne illness.

English

The present study was undertaken to investigate the microbiological quality of milk and artisanal dairy products sold in Ouagadougou markets. Forty-five (45) samples of milk and dairy products including raw milk, sour milk and yoghurt were collected and analyzed for aerobic mesophilic bacteria (AMB), total coliforms (TC), thermotolerant coliforms (TTC), Staphylococcus aureus (S. aureus), lactic acid bacteria (LAB) and yeasts and moulds (YM). The coliforms bacteria were then purified and identified by using API 20E system. The mean values of TC, TTC, S. aureus LAB and YM in the raw milk were 8.95, 6.43, 6.21, 4.83, 8.01 and 4.78 log cfu mL-1 respectively, In sour milk they were 10.44, 5.62, 4.12, 6.98, 7.38 and 3.75 log cfu mL-1 respectively, while in yoghurt samples the mean values of TC, TTC, S. aureus LAB and YM were 9.98, 5.77, 5.50, 6.12, 9.78 and 4.86 log cfu mL-1 respectively. The dominant coliforms isolated from raw milk, sour milk and yoghurt samples were found to be Escherichia coli, Klebsiella pneumonia and Enterobacter cloacae. Citrobacter spp was not found in any of the analyzed samples. The wide range of the samples analyzed did not comply with the standards quality. Thus it is necessary to set up an awareness program including capacity building of all the actors working in the sector of milk and dairy products. Key words: Raw milk, sour milk, yoghurt, microbiological quality, Ouagadougou.

Detection of Salmonella spp. with the BACTEC 9240 Automated Blood Culture System in 2008 - 2014 in Southern Iran (Shiraz): Biogrouping, MIC, and Antimicrobial Susceptibility Profiles of Isolates.

BackgroundHuman salmonellosis continues to be a major international problem, in terms of both morbidity and economic losses. The antibiotic resistance of Salmonella is an increasing public health emergency, since infections from resistant bacteria are more difficult and costly to treat.ObjectivesThe aims of the present study were to investigate the isolation of Salmonella spp. with the BACTEC automated system from blood samples during 2008 - 2014 in southern Iran (Shiraz). Detection of subspecies, biogrouping, and antimicrobial susceptibility testing by the disc diffusion and agar dilution methods were performed.Patients and MethodsA total of 19 Salmonella spp. were consecutively isolated using BACTEC from blood samples of patients between 2008 and 2014 in Shiraz, Iran. The isolates were identified as Salmonella, based on biochemical tests embedded in the API-20E system. In order to characterize the biogroups and subspecies, biochemical testing was performed. Susceptibility testing (disc diffusion and agar dilution) and extended-spectrum β-lactamase (ESBL) detection were performed according to the clinical and laboratory standards institute (CLSI) guidelines.ResultsOf the total 19 Salmonella spp. isolates recovered by the BACTEC automated system, all belonged to the Salmonella enterica subsp. houtenae. Five isolates (26.5%) were resistant to azithromycin. Six (31.5%) isolates with the disc diffusion method and five (26.3%) with the agar dilution method displayed resistance to nalidixic acid (minimum inhibitory concentration [MIC] > 32 μg/mL). All nalidixic acid-resistant isolates were also ciprofloxacin-sensitive. All isolates were ESBL-negative. Twenty-one percent of isolates were found to be resistant to chloramphenicol (MIC ≥ 32 μg/mL), and 16% were resistant to ampicillin (MIC ≥ 32 μg/mL).ConclusionsThe results indicate that multidrug-resistant (MDR) strains of Salmonella are increasing in number, and fewer antibiotics may be useful for treating S. enterica infections. Routine investigation and reporting of antibiotic MICs in patients presenting with Salmonella infections is suggested.

CTX-M-27 Producing Salmonella enterica Serotypes Typhimurium and Indiana Are Prevalent among Food-Producing Animals in China.

Salmonella spp. is one of the most important food-borne pathogens causing digestive tract and invasive infections in both humans and animals. Extended-spectrum β-lactamases (ESBLs) especially the CTX-M-type ESBLs are increasingly being reported worldwide and in China. These studies seldom focused on Salmonella isolates from food-producing animals. The aim of this study was to characterize the antimicrobial resistance profiles, serotypes and ESBLs and in particular, CTX-M producing Salmonella isolates from chickens and pigs in China. Salmonella isolates were identified by API20E system and polymerase chain reaction (PCR) assay; serotypes were determined using slide agglutination with hyperimmune sera; antimicrobial susceptibility was tested using the ager dilution method; the prevalence of ESBLs and PMQR genes were screened by PCR; CTX-M-producing isolates were further characterized by conjugation along with genetic relatedness and plasmid replicon type. In total, 159 Salmonella strains were identified, among which 95 strains were Salmonella enterica serovar Typhimurium, 63 strains were S. enterica serovar Indiana, and 1 strain was S. enterica serovar Enteritidis. All of these isolates presented multi-drug resistant phenotypes. Forty-five isolates carried blaCTX-M genes, the most common subtype was CTX-M-27(34), followed by CTX-M-65(7) and CTX-M-14(4). Most blaCTX-M genes were transmitted by non-typeable or IncN/IncFIB/IncP/IncA/C/IncHI2 plasmids with sizes ranging from 80 to 280 kb. In particular, all the 14 non-typeable plasmids were carrying blaCTX-M-27 gene and had a similar size. PFGE profiles indicated that CTX-M-positive isolates were clonally related among the same serotype, whilst the isolates of different serotypes were genetically divergent. This suggested that both clonal spread of resistant strains and horizontal transmission of the resistance plasmids contributed to the dissemination of blaCTX-M-9G-positive Salmonella isolates. The presence and spread of CTX-M, especially the CTX-M-27 in S. enterica serovars Typhimurium and Indiana from food-producing animals poses a potential threat for public health. Control strategies to limit the dissemination of these strains through the food chain are necessary.

Prevalence of Pathogenic Bacterial Isolates Infecting Wounds and their Antibiotic Sensitivity

Wounds infection by pathogenic bacteria and development of antibiotic resistance are the common post-surgical, medical challenges. Thus, the main objective of this work was to identify the common pathogenic bacteria infecting wounds and their resistance to antibiotic. Forty-one bacterial isolates belonging to the species Klebsiella pneumoniae, Enterobacter cloacae, Pseudomonas fluorescens, Providencia stuartii, Serratia rubidaea and Citrobacter freundii, infecting wounds were isolated from different hospitals in Cairo, Egypt. The isolates were biochemically identified by the API 20E system. The sensitivity of the forty-one identified bacterial isolates to twelve antibiotics was assessed by disc diffusion assay. Keywords Pathogenic bacterial; Wound infection; Multidrug resistance; Susceptibility patterns; Gram negative bacteria

Phenotypic and Genotypic Characterization of Pseudomonas Aeruginosa Isolates from Burn Patients

Background: Pseudomonas aeruginosa (P. aeruginosa) is an opportunistic pathogen that still develops life threatening infections in patients with immunological system defects like burns. It has been estimated that 75% of all deaths following burns are due to infections. Objectives: This study was designed to evaluate the prevalence and drug susceptibility profile of P. aeruginosa in patients admitted to a burns unit and to investigate the potential sources of infection in burn patients by genotyping using pulsed field gel electrophoresis (PFGE). Methodology: The study was performed in King Fahd Hospital (KFH), Almadinah Almunwarah during a ten month period. A total of 106 burn wound infection (BWI), 40 environmental and 10 health care workers (HCWs) samples were isolated then identified by conventional biochemical methods and API 20E system. Antibiotyping was performed by disc diffusion. Molecular relatedness of P. aeruginosa strains was investigated by pulsed field gel electrophoresis. Results: The most isolated organism was P. aeruginosa (31.6%), followed by Methicillin–Resistant Staphylococcus aureus (MRSA) (21%) and Acinetobacter baumannii (A.baumanii) (17.5). Colistin (100%) was the most effective drug against P. aeruginosa while the least effective drug was ticarcillin (48.1%). Typing of 27 isolates by PFGE revealed four pulso-types A, B, C and D and pulsotype B revealed 3 subtypes, subtypes B 1 , B 2 and B 3 . The isolates showed that 77.7% of isolates (from patients and environmental specimens) were genetically related. Also, Six patients had the same pulsotypes which were not detected in the environment. Conclusion: P.aeruginosa became one of the major concern in our hospital and implementation of infection control strategies is a major concern to avoid the spread of this threat. Environmental sources may have a significant role in spread of P. aeruginosa among hospitalized patients.

Matrix-assisted laser desorption ionization-time of flight mass spectrometry based identification of Edwardsiella ictaluri isolated from Vietnamese striped catfish (Pangasius hypothalamus).

Edwardsiella (E.) ictaluri is a major bacterial pathogen that affects commercially farmed striped catfish (Pangasius hypothalamus) in Vietnam. In a previous study, 19 strains of E. ictaluri collected from striped catfish were biochemically identified with an API-20E system. Here, the same 19 strains were used to assess the ability of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS; applied using a MALDI Biotyper) to conduct rapid, easy and accurate identification of E. ictaluri. MALDI-TOF MS could directly detect the specific peptide patterns of cultured E. ictaluri colonies with high (> 2.0, indicating species-level identification) scores. MALDI Biotyper 3.0 software revealed that all of the strains examined in this study possessed highly similar peptide peak patterns. In addition, electrophoresis (SDS-PAGE) and subsequent immuno-blotting using a specific chicken antibody (IgY) against E. ictaluri revealed that the isolates had highly similar protein profiles and antigenic banding profiles. The results of this study suggest that E. ictaluri isolated from striped catfish in Vietnam have homologous protein compositions. This is important, because it indicates that MALDI-TOF MS analysis could potentially outperform the conventional methods of identifying E. ictaluri.

Phenotypic and Genetic Diversity of Aeromonas Species Isolated from Fresh Water Lakes in Malaysia.

Gram-negative bacilli of the genus Aeromonas are primarily inhabitants of the aquatic environment. Humans acquire this organism from a wide range of food and water sources as well as during aquatic recreational activities. In the present study, the diversity and distribution of Aeromonas species from freshwater lakes in Malaysia was investigated using glycerophospholipid-cholesterol acyltransferase (GCAT) and RNA polymerase sigma-factor (rpoD) genes for speciation. A total of 122 possible Aeromonas strains were isolated and confirmed to genus level using the API20E system. The clonality of the isolates was investigated using ERIC-PCR and 20 duplicate isolates were excluded from the study. The specific GCAT-PCR identified all isolates as belonging to the genus Aeromonas, in agreement with the biochemical identification. A phylogenetic tree was constructed using the rpoD gene sequence and all 102 isolates were identified as: A. veronii 43%, A. jandaei 37%, A. hydrophila 6%, A. caviae 4%, A. salmonicida 2%, A. media 2%, A. allosaccharophila 1%, A. dhakensis 1% and Aeromonas spp. 4%. Twelve virulence genes were present in the following proportions—exu 96%, ser 93%, aer 87%, fla 83%, enolase 70%, ela 62%, act 54%, aexT 33%, lip 16%, dam 16%, alt 8% and ast 4%, and at least 2 of these genes were present in all 102 strains. The ascV, aexU and hlyA genes were not detected among the isolates. A. hydrophila was the main species containing virulence genes alt and ast either present alone or in combination. It is possible that different mechanisms may be used by each genospecies to demonstrate virulence. In summary, with the use of GCAT and rpoD genes, unambiguous identification of Aeromonas species is possible and provides valuable data on the phylogenetic diversity of the organism.

Isolation and Identification of Pseudomonas from Soil and different cases and study of susceptibility to antibiotics

85 samples were collected to investigate Pseudomonas sp. bacteria and study their resistance to antibiotics. The samples were included (18 specimen of wounds, 21 of burns, 23 specimen otitis media, 6 of urine tract infact, 3 of sputum) and soil samples . These cases were diagnosed using cultural and biochemical tests, the diagnosis was confirmed using the API 20E system. The results showed high isolation from burns and otitis media (23%) . The species was obtained Pseudomonas aeruginosa 49 (94.2%) followed by the bacteria Pseudomonas fluorescens 2 (3.8%), Pseudomonas oryzihabitans 1 (1.9%). Resistance of Pseudomonas sp. isolates to antibiotics were examined to 10 antibiotics, and isolates showed highest sensitivity to antibiotics Imipinem, Ciprofloxacin, Amikacin 98.1% and 92.3% and 90.4%, respectively. Isolates were showed high resistant (88.5%) to Cefotaxime .