Detection of Extended-Spectrum Beta-Lactamase-Producing Enterobacteriaceae in Desi Chickens in Andhra Pradesh

A total of 150 cloacal swabs were collected from desi chickens, 217 Enterobacteriaceae isolates were identified. The phenotypic antimicrobial resistance among Enterobacteriaceae was studied for 14 selected antibiotics by disc diffusion method. The selection of antibiotics was based on usage of antibiotics in commercial poultry farms and also based on priority of critically important antibiotics in humans. All Enterobacteriaceae isolates were subjected to multiplex PCR - I and II for detection of blaTEM, blaSHV and blaOXA genes and blaCTX-M, group 1 and 2 genes. Predominant β- Lactamase genes in gut microbiota of desi chicken include blaTEM (90.55%) followed by blaCTX-M group I (25.86%) and blaSHV (9.44%) genes. All the samples were found to be negative for blaOXA and blaCTX-M group 2 genes

Characterisation of Escherichia coli Recovered from Wild Animals Kept at Bikaner Zoo for Their Antibiotic Resistance

In the present study, a total of 41 E. coliisolates obtained from Boselaphus tragocamelus (5), Antelope cervicapra(18) and Gazella gazelle (18) maintained at Bikaner (Rajasthan) zoo were subjected to antibiogram determination against 15 antibiotics belonging to four different classes and were also genotyped for detecting presence of blaTEM,sul-2, strA and aadAgenes. Antibiogram study revealed highest efficacy of ciprofloxacin (90.2%) followed by nalidixic acid (75.6%) and chloramphenicol (68.4%) and high resistance to β lactam, Sulfamethoxazole and Aminoglycoside antibiotics. The overall presence of blaTEM, sul-2, strA and aadA genes was detected in 95.12%, 80.48%, 60.97% and 87.8% in isolates.

Prevalence of Extended Spectrum Beta Lactamase Genotypes in Klebsiella pneumonia from Respiratory Tract Infections at Tertiary Care Hospital.

Background: Extended Spectrum Beta Lactamases(ESBLs) are rapidly evolving group of β-lactamaseenzymes that are of particular concern to clinicians andepidemiologists. Most ESBLs have been evolved bygenetic mutation from blaTEM and blaSHV genes, andare well described in Klebsiella pneumoniae. Aim andObjective: To investigate the ESBL genotypes in K.pneumoniae isolates from Respiratory Tract Infections(RTIs). Material and Methods: Clinical isolates of K.pneumoniae were obtained from RTI -sputum samples.Antimicrobial susceptibility was determined by KirbyBauer disc diffusion method. ESBL was detectedphenotypically and multiplex Polymerase ChainReaction (PCR) specific for blaTEM, blaSHV andblaCTX-M genes was performed to identify genotypes.Results: During the 19 months period, a total of 212 ofK. pneumoniae were found from RTIs. Of these 212isolates, 60 isolates (28.3%) were ESBL producers byphenotypic method. Of these 212 isolates, 96 wererandomly selected for multiplex PCR for blaTEM,blaSHV and blaCTX-M genes. The findings ofmultiplex PCR showed that 24 isolates (25%)possessed blaTEM gene and only 4 isolates (4.1%)possessed each blaSHV and blaCTX-M gene alone.Isolates having both blaTEM+blaSHV genes were 20(20.8%), and both blaTEM+blaCTX-M genes were 12(12.5%); and isolate possessing all threeblaTEM+blaSHV+blaCTX-M genes were 20 (20.8%).The overall prevalence of blaTEM, blaSHV andblaCTX-M genes in this study was 79.1%, 45.8% and37.5% respectively. Imipenem was most effectiveantibiotic. Conclusion: Spread of ESBL producing K.pneumoniae is a major concern, as it causes limitationsto optimal treatment. Multiplex PCR can be used as arapid method to identify ESBL genotypes in K.pneumoniae. It will prove valuable for surveillance andestablishing the treatment line against drug resistantorganisms, thus saving precious time and resources. Inour study blaTEM genotype was most prevalent.

An emerging threat of ceftriaxone-resistant non-typhoidal salmonella in South India: Incidence and molecular profile

Background: Non-typhoidal Salmonella (NTS) infection is a serious public health problem globally. Although NTS infections are self-limited, antimicrobial therapy is recommended for severe infections and immunocompromised patients. Antimicrobial resistance (AMR) in these pathogens further limits its therapeutic options. Here, we report an incidence of ceftriaxone resistance in NTS over the past 9 years in a southern Indian region. Materials and Methods: Molecular mechanisms of resistance in ceftriaxone-resistant NTS have been tested by both phenotypic and molecular methods. Minimum inhibitory concentration was determined by the E-test and broth microdilution method. AMR gene markers of ?-lactamases such as AmpCs (blaMOX, blaCMY, blaDHA, blaFOX, blaACC and blaACT) and extended-spectrum ?-lactamases (ESBLs) (blaSHV, blaTEM, blaVEB, blaPER, blaCTXM-1like,blaCTXM-2like, blaCTXM-8like, blaCTXM-9like and blaCTXM-25like) were screened. The presence of IncH12 and IncI1 plasmid was also analysed. Results: The study reports a 5% prevalence of ceftriaxone resistance in NTS. The most common serogroup was Salmonella Group B followed by Salmonella Group E and Salmonella group C1/C2. The occurrence of blaCTX-M-1, blaTEM, blaCMY and blaSHV genes was observed in 54%, 54%, 48% and 3% of the isolates, respectively. Interestingly, few isolates carried dual resistance genes (ESBLs and AmpCs). IncH12 and IncI1 plasmid was identified in isolates carrying ESBL and AmpC genes, respectively. Conclusion: This study shows that ceftriaxone resistance is mainly mediated by ?-lactamases such as ESBL and AmpC. As the incidence of ceftriaxone resistance is rising gradually over the years, it is imperative to monitor the AMR in this species.

Phylogeny and Detection of blaTEM, blaSHV, blaCTX-M Genes in Escherichia coli Isolates from Patients with Urinary Tract Infections in Taif Hospitals, Saudi Arabia

Background: Urinary tract infections (UTIs) represent one of the most common diseases that are encountered in clinical practice and are caused mainly by Escherichia coli (E. coli). Aims: The objectives of this study were to identify and compare the blaTEM, blaSHV and blaCTX-M as marker of beta-lactamase genes in E. coli strains isolated from patients with UTIs collecting from King Abdul-Aziz hospital in Taif region, Saudi Arabia. Study Design: In vitro experimental and molecular study. Place and Duration of Study: Genetic engineering and biotechnology unit, Taif University, from September, 2016 to November, 2017. Methodology: Beta-lactame antibiotics are prescribed in most infectious disease including UTIs. Twenty one isolates identified as E. coli using microbial identification and confirmed by 16S rDNA. Results: These isolates were susceptible to Imipenem (100%), Ampicillin (90%) and Cefoxitin, but resistant to Cefepime (38%). Existance of selected bla-genes (blaTEM, blaSHV and blaCTX-M) were detected in the 21 isolates by PCR. Moreover, phylogeny tree was drawn based on 16S rDNA sequence. The results of this study show significant differences in susceptibility to different beta-lactam antibiotics among the bla-genes in E. coli isolates. Conclusion: Therefore, our findings instead of our data provide some new epidemiological information about the clonal nature of E. coli isolated from patients with UTIs in Taif region, KSA.

Recent Increase in the Incidence of TEM-135 β-Lactamase-harboring Neisseria gonorrhoeae in Korea

BACKGROUND: We investigated the molecular epidemiological characteristics and antimicrobial susceptibility pattern of penicillinase-producing Neisseria gonorrhoeae (PPNG) isolates to monitor the change in distribution of bla(TEM) in Korea. METHODS: We collected 804 PPNG isolates from diverse hospitals and clinics mainly located in Seoul, Korea, over a period of 11 years (2005–2015). Isolate susceptibility to seven antimicrobials was determined using the agar dilution test. The molecular epidemiological characteristics of the isolates were determined by Sanger sequencing of bla(TEM), N. gonorrhoeae multiantigen sequence typing (NG-MAST) and plasmid typing. RESULTS: Among 72 fully sequenced PPNG isolates, sixteen (22.2%) possessed TEM-135. All TEM-135 isolates had a common silent mutation (c.18C>T), which was previously unreported. We observed a pattern of continuous increase in the number of TEM-135 isolates since 2012. The median and 90% minimum inhibitory concentration of azithromycin were substantially lower in the TEM-135 group than in the non-PPNG and TEM-1 groups. All TEM-135 isolates showed different NG-MAST types and predominantly harbored Toronto/Rio (75%) plasmids. A comprehensive comparative analysis of PPNG with TEM-135 according to NG-MAST, plasmid type, and year of isolation revealed a wide distribution. CONCLUSIONS: The proportion of TEM-135 PPNG has continuously increased since 2012, in association with clonal spread. The difference at position 18 of the TEM-135 sequence can be interpreted as the existence of multiple clonal complexes. The possibility that TEM-135 was acquired via foreign plasmids requires careful follow-up and continuous monitoring of TEM-135 to ascertain whether it constitutes a step towards evolutionary change.

Study on the drug resistance of clinical strain of acinetobacter baumannii mediated by TEM-1 beta-lactamases and porin CarO / 天津医药

Objective To investigate the mechanism of drug resistance of sulbactam mediated by TEM-1 beta-lactamases and porin CarO in clinical strains of acinetobacter baumannii. Methods Twenty-four unrepeated clinical acinetobacter baumannii strains were divided into sensitive strain group (n=6) and insensitive strain group (n=18) by susceptibility testing to sulbactam.Antibiotics susceptibility test was carried out using the Kirby-Bauer method.BlaTEM-1and carO genes were amplified by PCR.Four blaTEM-1positive strains(A3,A5,1327 and C1)were selected,and their amplified products were sequenced.The quantitative real-time RT-PCR was used to analyze mRNA transcriptional levels of blaTEM-1 and carO genes. Results Resistant rates of the sensitive strain group for meropenem, imipenem, cefoperazone, ciprofloxacin,gentamicin and ampicillin were 1/6,2/6,3/6,1/6,5/6 and 5/6,and resistant rates of the insensitive strain group were 6/18,10/18,18/18,18/18 and 18/18.BlaTEM-1genes were amplified in 16 insensitive strains,and blaTEM-1was negative in sensitive strains. The carO genes were amplified in all 24 strains.There was no significative mutation in the 4 strains of blaTEM-1genes. The promoters of the strains A3, A5 and 1327 were P4, and C1 was P3. There was a positive correlation between the mRNA expression of blaTEM-1and the MIC value of sulbactam (rs=0.551, P=0.027). There was no difference in the mRNA expression of carO between the two groups.Conclusion The clinical strains are seriously resistant to antibiotics.The main resistance mechanism of clinical strains to sulbactam is the high mRNA expression of blaTEM-1,and the promoter may be one of the reasons of high expression of TEM-1.

Phenotypic and Molecular Characterization of Antibiotics Resistance E. Cloacae Isolates.

The present study aimed detecting and characterizing of β-lactamases producing E.cloacae isolated from different clinical sources in Hilla hospitals using phenotypic and molecular methods. A total of 308 samples were collected from two major hospitals at Hilla Province from October 2013 to April 2014. All isolates were tested biochemically, it was found that only 15 isolates from all isolates were belonging to Enterobactercloacae. All E. cloacae isolates were primarily screened for β-lactams resistance. Antibiotic susceptibility and minimum inhibitory concentration tests were performed using disk diffusion and agar dilution methods, respectively. The molecular study documented a widespread of Amp C genes among isolates of E. cloacae isolatesrepresented by 6/15(40%) positive isolates for Amp C primers. PCR assay revealed that prevalence rate of bla-TEM gene among tested isolates was 9(60%). followed by the bla-OXA gene was detected only in 3(20%).While bla-VEB gene and bla-SHV gene was not detected in any of the isolates. Some virulence factors of bacteria were also studied, and the results showed that all bacterial strains have capsule ,the results also also detected biofilm formation among isolates and the results revealed that 13(86%) of the isolates are biofilm former.

Penicillinase-producing Neisseria gonorrhoeae and its blaTEM-135 gene variants at several gonococcal antimicrobial surveillance sites in China：an epidemiological study / 中华皮肤科杂志

Objective To determine the prevalence of penicillinase-producing Neisseria gonorrhoeae(PPNG) and the distribution of blaTEM-135 gene variants in PPNG at several gonococcal antimicrobial surveillance sites in China, to compare N. gonorrhoeae multi-antigen sequence typing(NG-MAST)types of PPNG and its blaTEM-135 gene variants, and to assess the difference and association in NG-MAST types of blaTEM-135 gene variants among different regions. Methods A total of 572 N. gonorrhoeae isolates were collected at 6 gonococcal antimicrobial surveillance sites from Jiangsu, Shanghai, Zhejiang, Tianjin, Guangdong and Guangxi in 2012. After isolation, purification, and identification, cefalotin paper discs were used for detection of PPNG. DNA was extracted by QIAxtractor DX kits after cultivation of the PPNG strains. Then, mismatch amplification mutation assay (MAMA) PCR was performed to identify blaTEM-135 variants, and NG-MAST analysis to determine N. gonorrhoeae genotypes. Results Among the 572 N. gonorrhoeae strains, 38.1%(218/572) were identified as PPNG, and of the PPNG strains, 52.3% (114/218) were blaTEM-135 variants. The detection rate of PPNG at these surveillance sites from high to low was as follows: 51.7% (45/87, Zhejiang), 45.6%(36/79, Shanghai), 38.0% (78/205, Guangdong), 37.5% (12/32, Guangxi), 31.2% (24/77, Jiangsu) and 25.0%(23/92, Tianjin), and that of blaTEM-135 variants was as follows: 68.9%(31/45, Zhejiang), 58.3%(14/24, Jiangsu), 50.0%(39/78, Guangdong), 47.2%(17/36, Shanghai), 39.1%(9/23, Tianjin)and 33.3%(4/12, Guangxi). NG-MAST analysis showed that the ST2318, ST1768, ST1866, ST1053 and ST8726 types predominated among these bla TEM-135 variants, and a strong correlation was found between blaTEM-135 variants and some NG-MAST types, such as ST1768, ST1053 and ST8726 types. The distribution of NG-MAST types was significantly different between the surveillance site in Tianjin (in the Northern part of China) and the other sites (in the Southern part of China), but highly similar among the surveillance sites in Jiangsu, Zhejiang and Shanghai regions. Conclusions There is a high prevalence of PPNG and its blaTEM-135 variants at several gonococcal antimicrobial surveillance sites in China, with significant differences in NG-MAST genotype distribution of PPNG and its blaTEM-135 variants among different regions.

Molecular detection and antimicrobial resistance of Pseudomonas aeruginosa from houseflies (Musca domestica) in Iran

BackgroundPseudomonas aeruginosa is a common bacterium that can cause disease in humans and other animals. This study was conducted to screen for molecular detection and antimicrobial-resistant P. aeruginosa in Musca domestica in different locations in the Iranian provinces of Shahrekord and Isfahan.MethodsMusca domestica were captured by both manual and sticky trap methods, during the daytime, from household kitchens, cattle farms, animal hospitals, human hospitals, slaughterhouses and chicken farms at random locations in Shahrekord and Isfahan provinces of Iran, and subsequently transported to the laboratory for detection of P. aeruginosa. In the laboratory, flies were identified and killed by refrigeration in a cold chamber at −20 °C, then placed in 5 mL peptone water and left at room temperature for five hours before being processed. Pseudomonas isolates were preliminarily identified to genus level based on colony morphology and gram staining, and their identity was further confirmed by polymerase chain reaction.ResultsOverall blaTEM gene was recovered from 8.8 % (53/600) of the P. aeruginosa isolated from houseflies collected from the two provinces. A slightly higher prevalence (10.7 %; 32/300) was recorded in Shahrekord province than Isfahan province (7.0 %; 21/300). The locations did not differ statistically (p < 0.05) in bacterial prevalence in flies. Seasonal prevalence showed a significantly lower infection frequency during autumn.ConclusionsHouseflies are important in the epidemiology of P. aeruginosa infections.(AU)

Molecular detection and antimicrobial resistance of Pseudomonas aeruginosa from houseflies (Musca domestica) in Iran

Background Pseudomonas aeruginosa is a common bacterium that can cause disease in humans and other animals. This study was conducted to screen for molecular detection and antimicrobial-resistant P. aeruginosa in Musca domestica in different locations in the Iranian provinces of Shahrekord and Isfahan. Methods Musca domestica were captured by both manual and sticky trap methods, during the daytime, from household kitchens, cattle farms, animal hospitals, human hospitals, slaughterhouses and chicken farms at random locations in Shahrekord and Isfahan provinces of Iran, and subsequently transported to the laboratory for detection of P. aeruginosa. In the laboratory, flies were identified and killed by refrigeration in a cold chamber at −20 °C, then placed in 5 mL peptone water and left at room temperature for five hours before being processed. Pseudomonas isolates were preliminarily identified to genus level based on colony morphology and gram staining, and their identity was further confirmed by polymerase chain reaction. Results Overall blaTEM gene was recovered from 8.8 % (53/600) of the P. aeruginosa isolated from houseflies collected from the two provinces. A slightly higher prevalence (10.7 %; 32/300) was recorded in Shahrekord province than Isfahan province (7.0 %; 21/300). The locations did not differ statistically (p < 0.05) in bacterial prevalence in flies. Seasonal prevalence showed a significantly lower infection frequency during autumn. Conclusions Houseflies are important in the epidemiology of P. aeruginosa infections.

Distribution of beta-Lactamase Genes Among Carbapenem-Resistant Klebsiella pneumoniae Strains Isolated From Patients in Turkey

BACKGROUND: The emergence of carbapenem-resistant Klebsiella pneumoniae poses a serious problem to antibiotic management. We investigated the beta-lactamases in a group of carbapenem-resistant K. pneumoniae clinical isolates from Turkey. METHODS: Thirty-seven strains of K. pneumoniae isolated from various clinical specimens were analyzed by antimicrobial susceptibility testing, PCR for the detection of beta-lactamase genes, DNA sequencing, and repetitive extragenic palindronic (REP)-PCR analysis. RESULTS: All 37 isolates were resistant to ampicillin, ampicillin/sulbactam, piperacillin, piperacillin/tazobactam, ceftazidime, cefoperazone/sulbactam, cefepime, imipenem, and meropenem. The lowest resistance rates were observed for colistin (2.7%), tigecycline (11%), and amikacin (19%). According to PCR and sequencing results, 98% (36/37) of strains carried at least one carbapenemase gene, with 32 (86%) carrying OXA-48 and 7 (19%) carrying NDM-1. No other carbapenemase genes were identified. All strains carried a CTX-M-2-like beta-lactamase, and some carried SHV- (97%), TEM- (9%), and CTX-M-1-like (62%) beta-lactamases. Sequence analysis of bla(TEM) genes identified a bla(TEM-166) with an amino acid change at position 53 (Arg53Gly) from bla(TEM-1b), the first report of a mutation in this region. REP-PCR analysis revealed that there were seven different clonal groups, and temporo-spatial links were identified within these groups. CONCLUSIONS: Combinations of beta-lactamases were found in all strains, with the most common being OXA-48, SHV, TEM, and CTX-M-type (76% of strains). We have reported, for the first time, a high prevalence of the NDM-1 (19%) carbapenemase in carbapenem-resistant K. pneumoniae from Turkey. These enzymes often co-exist with other beta-lactamases, such as TEM, SHV, and CTX-M beta-lactamases.

Multiple Antimicrobial Resistance in Vibrio spp Isolated from River and Aquaculture Water Sources in Imo State, Nigeria.

Aim: To study multiple antimicrobial resistances in Vibrio spp. isolated from river and aquaculture water sources in Imo State Nigeria. Methodology: A total of 157 Vibrio isolates from river and aquaculture water sources were analysed for multiple antimicrobial resistance during a 6 month period. Antimicrobial resistance profile was determined by the Kirby-Bauer technique, while the phenotypic expression of β-lactamase production was performed by the double disk diffusion method. PCR was used to screen isolates for the presence of β-lactamase resistance genes. Results: The isolates from river water expressed high resistance rates (81.3 to 97.8%) to the following antimicrobials: mezlocillin, doxycycline, tetracycline, carbenicillin and ampicillin, while resistance rate to kanamycin was moderate at 40.9%. Resistance rates for the aquaculture water Isolates were also high for the same antibiotics as the river water isolates, while resistance rate to kanamycin was low to moderate at 32.8%. Phenotypic screening of isolates for ESβL production showed the isolates were resistant to β-lactam antimicrobials and the β-lactamase inhibitor of amoxicillin/clavulnic acid combination. Gel electrophoresis of PCR products showed amplification for blaTEM of size 964bp. Conclusion: Results showed the presence of highly resistant Vibrio isolates from the sampled environmental sources. The presence of resistance markers among the isolates in this study infers that they could be agents of transfer of resistance to other bacterial pathogens found in river and aquaculture water.

Prevalence of blaTEM , blaSHV and blaCTX-M genes in clinical isolates of Escherichia coli and Klebsiella pneumoniae from Northeast India.

Aim: This study was carried out to determine the presence of blaTEM, blaSHV and blaCTX-M genes in extended-spectrum β-lactamase (ESBL) producing Escherichia coli (E. coli) and Klebsiella pneumoniae (K. pneumoniae) at a tertiary care referral hospital in Northeast India. Materials and Methods: A total of 270 E. coli and 219 K. pneumoniae isolates were recovered during the period between August 2009 and July 2010. Kirby-Bauer disk diffusion method was performed to determine the antibiotic resistance pattern. Screening and phenotypic confi rmatory test for ESBL production were performed using standard disc diffusion methods. Each of the initial ESBL screening test isolate was investigated for the presence of blaTEM, blaSHV and blaCTX-M genes via polymerase chain reaction (PCR) using gene-specifi c primers. Results: P henotypic confi rmatory test able to detect ESBL production in 73.58% of E. coli and 67.24% of K. pneumoniae. However, PCR amplifi cation showed the presence of one or more ESBL genes in each of the initial ESBL screening positive isolate. Among three ESBL genotypes, the most prevalent genotype was found to be blaCTX-M in E. coli (88.67%) and blaTEM in K. pneumoniae (77.58%) ESBL producing isolates. Majority of ESBL producing isolates possess more than one ESBL genes. Conclusion: T his study constituted a primer report on high prevalence of blaTEM and blaCTX-M genes in ESBL producing isolates of E. coli and K. pneumoniae and denotes the need of more extensive studies on these antibiotic genes to determine the magnitude of the problem of antibiotic resistance exiting in this locality.

Molecular detection of ESBLs production and antibiotic resistance patterns in Gram negative bacilli isolated from urinary tract infections.

Background: β-lactam resistance is more prevalent in Gram negative bacterial isolates worldwide, particularly in developing countries. In order to provide data relating to antibiotic therapy and resistance control, routine monitoring of corresponding antibiotic resistance genes is necessary. Aims: The aim of this study was the characterization of β-lactam resistance genes and its plasmid profi le in bacteria isolated from urinary tract infection samples. Materials and Methods: In this study, 298 Gram negative bacteria isolated from 6739 urine specimens were identifi ed by biochemical standard tests. Antimicrobial susceptibility testing was performed by the disk diffusion method. Extendedspectrum β-lactamase (ESBL)-producing strains were also detected by the double-disk synergy test. The presence of blaTEM and blaSHV genes in the strains studied was ascertained by polymerase chain reaction. Results: Of all Gram negative bacteria, Escherichia coli (69.1%) was the most common strain, followed by Klebsiella sp. (12.1%), Enterobacter sp. (8.4%), Proteus sp. (4.4%), Citrobacter (4%) and Pseudomonas sp. (2%). The most antibiotic resistance was shown to tetracycline (95.16%), nalidixic acid (89.78%) and gentamycin (73.20%) antibiotics. Among all the strains tested, 35 isolates (11.75%) expressed ESBL activity. The prevalence of TEM and SHV positivity among these isolates was 34.29%, followed by TEM (31.43%), TEM and SHV negativity (20.0%) and SHV (14.29%), respectively. Conclusions: Regular monitoring of antimicrobial drug resistance seems necessary to improve our guidelines in the use of the empirical antibiotic therapy.

Genes blaTEM, blaSHV y blaCTX-M en enterobacterias productoras de b-lactamasas de espectro extendido aisladas de pacientes con infección intrahospitalaria / blaTEM, blaSHV y blaCTX-M genes in extended-spectrum b-lactamases producing enterobacterias isolated from patients with hospital-acquired infections

El objetivo de este estudio fue identificar los genes blaTEM, blaSHV y blaCTX-M en aislados clínicos de enterobacterias productoras de b-lactamasas de espectro extendido (BLEE), recolectadas entre septiembre y noviembre de 2005. Además de la resistencia a las cefalosporinas de tercera generación, los aislados también mostraron resistencia a cloranfenicol (59,2%) amikacina (37,0%) y gentamicina (40,7%) y se mostraron sensibles a imipenem y meropenem. Nueve cepas lograron transferir la resistencia a las cefalosporinas de tercera generación, así como la producción de BLEE. En los aislados clínicos se detectaron los genes blaSHV, blaTEM y blaCTX-M, donde los tipos blaTEM-1, blaSHV-1, blaSHV-5 blaSHV-5-2a y blaCTX-M-1 fueron los prevalentes; mientras que en las transconjugantes sólo se detectaron blaTEM-1, blaSHV-5 y blaSHV-5-2a. Se identificaron en total siete tipos de genes, de los cuales cinco eran codificantes de enzimas tipo BLEE, lo que demuestra que en el centro hospitalario la resistencia a las cefalosporinas de tercera generación es debida a diversas enzimas.

The objective of the present investigation was to identify the blaTEM, blaSHV and blaCTX-M genes on extended-spectrum b-lactamases (ESBL) producing Enterobacteriaceae from clinical isolates, collected between September and November 2005. In addition to third-generation cephalosporin resistance, the isolates also showed resistance to chloramphenicol (59.2%), amikacin (37.0%) and gentamicin (40.7%), and demonstrated sensitivity to imipenem and meropenem. Nine strains were capable of transferring third-generation cephalosporin resistance, as well as the production of ESBL. In the clinical isolates, the genes blaSHV, blaTEM and blaCTX-M were detected, being more prevalent the types blaTEM-1, blaSHV-1, blaSHV-5 blaSHV-5-2a and blaCTX-M-1; while in the trans-conjugated only blaTEM-1, blaSHV-5 y blaSHV-5-2a were found. In total, seven types of genes were identified, five of which were codifying genes for ESBL-type enzymes. This demonstrates that in the hospital center, resistance to third-generation cephalosporin is mediated by several enzymes.

Antimicrobial resistance and frequency of BlaTEM in Escherichia coli isolated from non-diarrheic and diarrheic piglets / 대한수의학회지

Antimicrobial resistance is one of the most concerns in pig industry. Escherichia (E.) coli have been used for the indicator to monitor the antimicrobial resistance. In this study, 321 E. coli from diarrheic and non-diarrheic piglets were tested for antimicrobial resistance and frequency of Bla TEM. In non-diarrheic piglets, they were resistant to oxytetracycline (93%), streptomycin (92%) and sulfadiazine (90%) but susceptible to ceftiofur (99%), colistin (97%), and enrofloxacin (82%). The isolates from diarrheic piglets were resistant to enrofloxacin (72.9%), ceftiofur (17.6%), and colistin (11.3%), whereas the resistance was 1%, 18% and 3% in case of non-diarrheic piglets, respectively. The resistance for amoxicillin/clavulanic acid (54.1%) and ceftiofur (22%) was high in isolates from post-weaning piglets. The resistance for colistin was 15.2% in nursery piglets. Seventy-three percent of isolates from diarrheic piglets showed high multidrug resistance profile (more than 13 antimicrobials) compared to those from non-diarrheic pigs in which 71% of isolates showed moderate multidrug resistance profile (7 to 12 antimicrobials). The frequency of BlaTEM in E. coli from non-diarrheic and diarrheic piglets was 57% and 69%, respectively. The results might provide the basic knowledge to establish the strategies for treatment and reduce antibiotic resistance of E. coli in piglets.

Characterization of Vibrio Parahaemolyticus isolated from oysters and mussels in São Paulo, Brazil / Caracterização de Vibrio parahaemolyticus isolados de ostras e mexilhões em São Paulo, Brasil

Vibrio parahaemolyticus is a marine bacterium, responsible for gastroenteritis in humans. Most of the clinical isolates produce thermostable direct hemolysin (TDH) and TDH-related hemolysin (TRH) encoded by tdh and trh genes respectively. In this study, twenty-three V. parahaemolyticus, previously isolated from oysters and mussels were analyzed by PCR using specific primers for the 16S rRNA and virulence genes (tdh, trh and tlh) and for resistance to different classes of antibiotics and PFGE. Nineteen isolates were confirmed by PCR as V. parahaemolyticus. The tlh gene was present in 100 percent of isolates, the tdh gene was identified in two (10.5 percent) isolates, whereas the gene trh was not detected. Each isolate was resistant to at least one of the nine antimicrobials tested. Additionally, all isolates possessed the blaTEM-116 gene. The presence of this gene in V. parahaemolyticus indicates the possibility of spreading this gene in the environment. Atypical strains of V. parahaemolyticus were also detected in this study.

Vibrio parahaemolyticus é uma bactéria marinha, responsável por gastroenterite em humanos. A maioria dos isolados clínicos produzem hemolisina termoestável direta (TDH) e hemolisina TDH-relacionada (TRH) codificadas por genes tdh e trh, respectivamente. Neste estudo, vinte e três V. parahaemolyticus, previamente isolados de ostras e mexilhões foram analisados por PCR utilizando indicadores específicos para o gene 16S rRNA, genes de virulência (tdh, trh e tlh), resistência a diferentes classes de antibióticos, e PFGE. Dezenove isolados foram confirmados por PCR, como V. parahaemolyticus. O gene tlh estava presente em 100 por cento dos isolados, o gene tdh foi identificado em dois (10,5 por cento) dos isolados, enquanto que o gene trh não foi detectado. Cada isolado foi resistente a pelo menos um dos nove antibióticos testados. Além disso, todos os isolados apresentaram resultado positivo para o gene blaTEM-116. A presença deste gene em V. parahaemolyticus indica a possibilidade de propagação desse gene no ambiente. Cepas atípicas de V. parahaemolyticus foram também detectadas neste estudo.

Presence of blaTEM-116 gene in environmental isolates of Aeromonas hydrophila and Aeromonas jandaei from Brazil

It is known that Aeromonas spp. possess different chromosomal â-lactamase genes. Presence and phenotypic expression of blaTEM, blaSHV, and blaCTX-M ESBL-encoding genes were investigated in environmental water isolates of Aeromonas hydrophila and Aeromonas jandaei. Presence of blaSHV and blaCTX-M genes was not observed, and blaTEM gene was verified in 91 percent of the isolates. Sequencing of 10 fragments showed the occurrence of blaTEM-116.

Some etiological and clinical aspects of pyelonephritis and antibiotic resistance of pathogens causing children’s pyelonephritis in Mongolia / Монголын Анагаах Ухаан

IntroductionPyelonephritis is generally the result of an ascending infection of the urinary tract (reflux is seen in 30- 50% of affected children) most common organism is Escherichia coli (Heiberger R.E., 2006). The bacteria that are most likely to cause pyelonephritis are those that normally occur in the feces. Pathogens isolated from patients with pyelonephritis include both gram positive and gram negative bacteria. R.E.Neiberger reported that of gram negative bacteria E.coli, Klebsielle, Enterobacter, Proteus, Pseudomonas and of gram positive bacteria Enterococcus, Staphylococcus saphrophiticus, group B.Streptococcus were prevalent in children with pyelonephritis. In another study conducted at the Pennsylvania University, USA, 70-95% of causes were related with E.coli, Proteus, Klebsiella, Pseudomonas, Staphylococcus saphrophiticus and Еnterobacter (Sammuel Baron, 2004). In the Indian study E.coli accounted for 88.1%, Pseudomonas aeruginosa for 7.1% and Klebsiella was in 4.8% of cases (Namalwar B.R., Vijayakumar M., Janani Sankar., Ramnath B and Prahlad N., 2004). Serotypes 01; 02; 04; 06; 07 were considered as most prevalent among E.coli causes of UTI (Sammuel Baron, 2004). Escherichia coli causes about 85% of acute bladder and kidney infections in patient with no obstruction or history of surgical procedures. Proteus, Klebsiella, Enterobacter or Pseudomonas are other common causes of infection(Namalwar B.R., Vijayakumar M., Janani Sankar., Ramnath B and Prahlad N., 2004). Once these organisms enter the urinary tract, they cling to the tissues that line the tract and multiply in them (Parveen J Kumar Michael L Clark Clinical medicine. London, 2003). Pyelonephritis is characterized by tubulo-interstitial inflammation, hyperaemia and oedema. Affected patients present with the abrupt onset of fever and chills, constant dull flank pain, and symptoms of cystitis (dysuria, frequency, urgency). Urinalysis usually procedures microabscess. Urine cultures will frequency be negative in the setting of haematogenousinfection (Namalwar B.R., Vijayakumar M., Janani Sankar., Ramnath B and Prahlad N., 2004). Asymptomatic infections of the urinary tract or Asymptomatic bactreuria are common. In childgood, about 1 percent of girls have asymptomatic bacteruria (Namalwar B.R., Vijayakumar M., Janani Sankar., Ramnath B and Prahlad N., 2004). In Mongolia females aged between 20-40 years old accounted for 60.3% of all patients with chronic pyelonephritis. In cases of bacteriuria E.Coli was 74.05%, with highest sensitivity to claforan (91.25%) and ciprofloxacin (90.65%) (B.Selengee, 2003). Last 5 years (2004-2008) 3709 children were treated in Nephrological department of Mathernal and Children’s Scientific Center, but between them 741(19.97%) children were illned by pyelonephritis. However no study was conducted on bacterial causes of pyelonephritis in children in Mongolia. Purpose:The main goal of study was to indicate the etiological and clinical specialaty of children’s pyelonephritis. Especially, to indicate the prevalently occurred type and the etiological bacterial groups also the clinical specialaty of children’s pyelonephritis. To study microbiological spectrum of pathogens causing pyelonephritis in Mongolian children, sensitivity of them to antimicrobial agent, mechanisms of their resistance and to identify genes related with microbial pathogenity.Materials and Мethods:My study covered 254 children wich illned by pyelonephritis, these were treated in Nephrological department of Mathernal and Children’s Scientific Center in last 5 years (2004-2008). From patient’s history we also used the clinical laboratorial analysis results such as biochemical, blood count and urine analysis. We used retrospective and prospective statistical methods for study on pathient’s history of children wich sickned by pyelonephritis and bacteriological method for indicating groups of bacteria. We collected information about patients by questionary chart. During study we also used electro-sonography(echo), Xray, radio-isotope, computer-tomography and reno-vascular angiography methods. Antibiotic susceptibility testing covered total 212 bacterial species to 19 kind of antibiotics by disk diffusion(DD) and minimium inhibitory concentration(MIC) methods. Strains were identified by API tests strip (BioMerieux, France) and antibiotic susceptibility was determined by disk diffusion method according to the ELSI recommendations. Types of the beta lactamase were determined by PCR using the blaTEM, blaSHV and blaCTX-M specific primer. 1080 bp, 400 bp and 570 bp PCR product were interpreted as positive for blaTEM, blaSHV and blaCTX-M genes. WHONET5.1 programm was used for the analysis of susceptibility testing according to the WHO recommendation. Pathogens from 1 month consequent urine culture of patients treated at the pediatric nephrology, urology units and outpatient clinic at the MCHRC and patients from nephrology and urology departments at the CCH were isolated using disk diffusion test and minimum inhibitory concentration. Bacterial resistance and pathogenity related genes were identified using PCR method.Results:There were the girls prevalently sicked by pyelonephritis (n=181 or 71.25%) than boys (n=73 or 28.74%) and the primary (n=155 or 61.02%) and secondary chronic pyelonephritis (n=99 or 38.97%) also was prevalently occurred. During pyelonephritis are occurring following clinical symptoms these are high temperature, edema eyelid, loss of appetite, sludged urinate, skin dryness, womiting and nicthuria. By our study the main clinical symptoms of children’s pyelonephritis are high temperature (58.55%), back pain (51.0%), edema of eyelid (34.42%) and polyuria (25.79%). Also the main influencing factors for inducing children’s pyelonephritis are hyphoplasia (33.3%), hydronephrosis (19.23%), single kidney (15.38%) and vesico-uretheral reflux (6.41%). Study was conducted on 110 positive cultures (E.coli-52, Citrobacter spp.-8, E.cloacae-10, Proteus spp.-7, K.pneumoniae-7, P.aeruginosae-22 ба A.baumennii-2, S.aureus-1, Enterococci-1). The bacterial sensitivity and resistance to antimicrobial agents differed between various pathogens. From Gram negative oxydase negative Enterobacteriaceae by PCR testing, 19 isolates contained blaTEM types, 11 strains contained blaSHV types and 16 strains contained blaCTX-M type genes. On the study of PCR the virulence gene “aer” were determined 50% of E.coli strains. blaTEM type resistance can divided to 5 groups, from them 2 groups had the resistance to single beta-lactamic antibiotics, but 3 groups had to both beta-lactamic and aminoglycoside antibiotics.blaSHV type resistance had 2 groups antibiotics, these were beta-lactamic and aminoglycoside antibiotics. blaCTX-M type resistance had 2 groups, these were beta-lactamic and aminoglycoside antibiotics. Both the blaTEM and blaSHV type co-resistance had 2 groups, these were CEP and CXA antibiotics. blaTEM and CTX-M type resistance can divided to 4 groups, from them 1 group had the resistance to single beta-lactamic antibiotics, other 3 groups had to aminoglycoside antibiotics. Both the blaSHV and CTX-M type co-resistance had 2 groups, these were lactamic and aminoglycoside antibiotics. blaTEM, SHV and CTX-M type co-resistance can divided to 3 groups, these were also beta-lactamic and aminoglycoside antibiotics. On the study of PCR the virulence “aer (aerobacterin) gene” expression was identified in 50% of E.coli strains.Conclusion:There were the girls (181 cases or 71.25%) prevalently sick by pyelonephritis than boys(73 cases or 28.74%) was prevalently occurred.In Mongllia the primary pyelonephritis(155 cases or 61.02%) was occurred prevalently than secondary chronic pyelonephritis( 99 cases or 38.97%). The gasteroenterological and toxic symptoms prevalently occurred in clinical symptoms, but later kidney and urinary tract symptoms noticed.In Mongolia the main influencing factors for inducing children’s pyelonephritis are hyphoplasia (33.3%), hydronephrosis (19.23%), single kidney (15.38%) and vesico-uretheral reflux (6.41%).The prevalent pathogen causing pyelonephritis in Mongolia is E.coli. Other causes include Enterobacter spp., Klebsilla spp., Proteus spp., P.aeruginosa, Aceinobacter baumenii and S.aureus. The virulence gene “aer” were determined 50% of E.coli strains. The most prevalent resistance was resistance to beta-lactam, aminoglycoside, tetracycline and trimethoprim sulfometohoxazole. Resistance to beta-lactamic antibiotics was related with blaSHV, blaTEM, blaCTX-M gene experessions in 25-40% of cases. blaTEM, blaSHV and bla CTX-M type of the beta-lactamase were determined 25-40% of Gram negative oxidase negativebacilli. Pathogenic “aer” gene was identified in 50% of E.coli in Mongolia.