Evaluation of the antibacterial activity of Weissella confusa K3 cell-free supernatant against extended-spectrum βeta lactamase (ESBL) producing uropathogenic Escherichia coli U60. - Related Documents




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209401.0000Evaluation of the antibacterial activity of Weissella confusa K3 cell-free supernatant against extended-spectrum βeta lactamase (ESBL) producing uropathogenic Escherichia coli U60. Different strategies have been approved for controlling extended-spectrum βeta lactamase (ESBL) producing uropathogenic bacteria. The antibacterial activity of Lactic acid bacteria (LAB) is an effective strategy due to its probiotic characteristics and beneficial effects on human health. The antibiotic susceptibility test, disk diffusion method, and double disc synergy test indicated that five enteric uropathogenic isolates were ESBL producers during the present study. They recorded diameters of inhibition zones as ≤ 18, ≤ 8, ≤ 19, and ≤ 8 mm against cefotaxime (CTX), ceftazidime (CAZ), aztreonam (ATM), and ceftriaxone (CRO). Genotypically, bla(TEM) genes are the most common, with (100 %) occurrence in all the five enteric tested uropathogens, followed by bla(SHV) and bla(CTX) genes (60 %). In addition, out of 10 LAB isolates from dairy products, the CFS of isolate no. K3 had high antibacterial activity against the tested ESBLs, especially no. U60, with a MIC of 600 µl. Additionally, the MIC and sub-MIC of K3 CFS inhibited the production of antibiotic-resistant bla (TEM) genes of U60. Analyzing the 16S rRNA sequence confirmed that the most potent ESBL-producing bacteria (U60) and LAB (K3) isolates were identified as Escherichia coli U60.1 and Weissella confuse K3 with accession numbers MW173246 and MW173299.1, respectively, in GenBank.202336873575
146610.9992Antibiotic resistance and genotype of beta-lactamase producing Escherichia coli in nosocomial infections in Cotonou, Benin. BACKGROUND: Beta lactams are the most commonly used group of antimicrobials worldwide. The presence of extended-spectrum lactamases (ESBL) affects significantly the treatment of infections due to multidrug resistant strains of gram-negative bacilli. The aim of this study was to characterize the beta-lactamase resistance genes in Escherichia coli isolated from nosocomial infections in Cotonou, Benin. METHODS: Escherichia coli strains were isolated from various biological samples such as urine, pus, vaginal swab, sperm, blood, spinal fluid and catheter. Isolated bacteria were submitted to eleven usual antibiotics, using disc diffusion method according to NCCLS criteria, for resistance analysis. Beta-lactamase production was determined by an acidimetric method with benzylpenicillin. Microbiological characterization of ESBL enzymes was done by double disc synergy test and the resistance genes TEM and SHV were screened by specific PCR. RESULTS: ESBL phenotype was detected in 29 isolates (35.5%). The most active antibiotic was imipenem (96.4% as susceptibility rate) followed by ceftriaxone (58.3%) and gentamicin (54.8%). High resistance rates were observed with amoxicillin (92.8%), ampicillin (94%) and trimethoprim/sulfamethoxazole (85.7%). The genotype TEM was predominant in ESBL and non ESBL isolates with respectively 72.4% and 80%. SHV-type beta-lactamase genes occurred in 24.1% ESBL strains and in 18.1% of non ESBL isolates. CONCLUSION: This study revealed the presence of ESBL producing Eschericiha coli in Cotonou. It demonstrated also high resistance rate to antibiotics commonly used for infections treatment. Continuous monitoring and judicious antibiotic usage are required.201525595314
112820.9992Molecular 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 profile in bacteria isolated from urinary tract infection samples. MATERIALS AND METHODS: In this study, 298 Gram negative bacteria isolated from 6739 urine specimens were identified by biochemical standard tests. Antimicrobial susceptibility testing was performed by the disk diffusion method. Extended-spectrum 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.201424943757
116130.9992Detection of extended-spectrum β-lactamase-producing Escherichia coli genes isolated from cat rectal swabs at Surabaya Veterinary Hospital, Indonesia. BACKGROUND AND AIM: Escherichia coli causes a bacterial illness that frequently affects cats. Diseases caused by E. coli are treated using antibiotics. Because of their proximity to humans, cats possess an extremely high risk of contracting antibiotic resistance genes when their owners touch cat feces containing E. coli that harbor resistance genes. This study was conducted to identify multidrug-resistant E. coli and extended-spectrum β-lactamase (ESBL)-producing genes from cat rectal swabs collected at Surabaya City Veterinary Hospital to determine antibiotic sensitivity. MATERIALS AND METHODS: Samples of cat rectal swabs were cultured in Brilliant Green Bile Lactose Broth medium and then streaked on eosin methylene blue agar medium for bacterial isolation, whereas Gram-staining and IMViC tests were conducted to confirm the identification results. The Kirby-Bauer diffusion test was used to determine antibiotic sensitivity, and the double-disk synergy test was used to determine ESBL-producing bacteria. Molecular detection of the genes TEM and CTX-M was performed using a polymerase chain reaction. RESULTS: Based on morphological culture, Gram-staining, and biochemical testing, the results of sample inspection showed that of the 100 cat rectal swab samples isolated, 71 (71%) were positive for E. coli. Furthermore, 23 E. coli isolates (32.39%) demonstrated the highest resistance to ampicillin. Four isolates were confirmed to be multidurg-resistant and ESBL-producing strains. Molecular examination revealed that three E. coli isolates harbored TEM and CTX-M. CONCLUSION: In conclusion, pet owners must be educated on the use of antibiotics to improve their knowledge about the risks of antibiotic resistance.202337859949
98740.9992Characterization of Multidrug Resistant Extended-Spectrum Beta-Lactamase-Producing Escherichia coli among Uropathogens of Pediatrics in North of Iran. Escherichia coli remains as one of the most important bacteria causing infections in pediatrics and producing extended-spectrum beta-lactamases (ESBLs) making them resistant to beta-lactam antibiotics. In this study we aimed to genotype ESBL-producing E. coli isolates from pediatric patients for ESBL genes and determine their association with antimicrobial resistance. One hundred of the E. coli isolates were initially considered ESBL producing based on their MIC results. These isolates were then tested by polymerase chain reaction (PCR) for the presence or absence of CTX, TEM, SHV, GES, and VEB beta-lactamase genes. About 30.5% of isolated E. coli was ESBL-producing strain. The TEM gene was the most prevalent (49%) followed by SHV (44%), CTX (28%), VEB (8%), and GES (0%) genes. The ESBL-producing E. coli isolates were susceptible to carbapenems (66%) and amikacin (58%) and showed high resistance to cefixime (99%), colistin (82%), and ciprofloxacin (76%). In conclusion, carbapenems were the most effective antibiotics against ESBl-producing E. coli in urinary tract infection in North of Iran. The most prevalent gene is the TEM-type, but the other resistant genes and their antimicrobial resistance are on the rise.201526064896
98450.9992Genetic characterization of TEM-type ESBL-associated antibacterial resistance in Enterobacteriaceae in a tertiary hospital in Ghana. BACKGROUND: Antibiotic resistance due to the presence of extended-spectrum beta-lactamases (ESBLs) among Enterobacteriaceae is a worldwide problem. Data from Ghana regarding this resistance mechanism is limited. This study was designed to investigate the presence of TEM-type ESBL genes, their locations and their conjugabilities in clinical isolates of enterobacteria collected from the Korle-Bu Teaching Hospital in Ghana. METHODS: Study isolates were characterized with respect to ESBL phenotype, TEM-type ESBL gene detection, location of the ESBL gene(s) and conjugability of the ESBL phenotype using nalidixic acid-resistant Escherichia coli K-12 as recipient. Phenotyping was by Kirby Bauer disk diffusion using cefpodoxime, ceftazidime, cefotaxime and their combinations with clavulanate. Gene detections were by PCR using blaTEM primers. RESULTS: Overall, 37.96 % of 137 clinical isolates showed ESBL phenotype. The ESBLs occurred mostly in Klebsiella spp. (42.3 %) and then Escherichia coli (34.6 %). The TEM gene was detected in 48.1 % of ESBL-positive isolates and was determined to be plasmid-borne in 24 of 25 blaTEM detections. Overall, 62.7 % of TEM-producing isolates transferred the ESBL phenotype by conjugation. CONCLUSIONS: The results highlight the presence of TEM-type ESBLs in the Korle-Bu Teaching Hospital and show considerable risk of environmental contamination through the urine of infected persons. An inhibition zone chart was generated which indicates the possible presence of complex beta-lactamase types. The data points to the fact that the ESBL-producing bacteria may disseminate this resistance mechanism via conjugation.201627145868
99760.9991Prevalence and antibacterial resistance patterns of extended-spectrum beta-lactamase producing Gram-negative bacteria isolated from ocular infections. PURPOSE: Extended-spectrum beta-lactamases (ESBLs) mediated resistance is more prevalent worldwide, especially among Gram-negative bacterial isolates, conferring resistance to the expanded spectrum cephalosporins. As limited data were available on the prevalence of ESBLs in this area, the current study was undertaken to determine the prevalence, antibacterial resistance patterns, and molecular detection and characterization of ESBL encoding resistance genes among ocular Gram-negative bacterial isolates from ocular infections. MATERIALS AND METHODS: A prospective study was done on 252 ocular Gram-negative bacterial isolates recovered from ocular infections during a study period from February 2011 to January 2014. All isolates were subjected to detection of ESBLs by cephalosporin/clavulanate combination disc test and their antibacterial resistance pattern was studied. Molecular detection and characterization of ESBL encoding blaTEM -, blaSHV , blaOXA -, and blaCTX-M (phylogenetic groups 1, 2, 9, and 8/25) resistance genes by multiplex polymerase chain reaction and DNA sequence analysis. RESULTS: Of all Gram-negative bacteria, Pseudomonas aeruginosa (44%) was the most common strain, followed by Enterobacter agglomerans and Klebsiella pneumoniae each (10%). Among the 252, 42 (17%) were ESBL producers. The major source of ESBL producers were corneal scraping specimens, highest ESBL production was observed in P. aeruginosa 16 (38%) and Escherichia coli 7 (16.6%). Among ESBL-producing genes, the prevalence of blaTEM -gene was the highest (83%) followed by blaOXA -gene (35%), blaSHV -gene (18.5%), and blaCTX-M-1 -gene (18.5%) alone or together. CONCLUSION: The higher rate of prevalence of ESBLs-encoding genes among ocular Gram-negative bacteria is of great concern, as it causes limitation to therapeutic options. This regional knowledge will help in guiding appropriate antibiotic use which is highly warranted.201627221683
105970.9991Dissemination and phenotypic characterization of ESBL-producing Escherichia coli in Indonesia. BACKGROUND: The alarming rise in infections caused by extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli in animals and humans poses a serious threat due to its escalating antibiotic resistance. Unveiling this problematic bacteria's prevalence and resistance patterns in animals is crucial for formulating effective control strategies and safeguarding public health. AIM: The purpose of this study was to analyze the expression of three main genes: blaCTX-M, blaSHV, and blaTEM, in ESBL-producing E. coli isolates from The Research Center for Veterinary Science and the National Research and Innovation Agency. Moreover, their resistance profiles against various antibiotics should be systematically evaluated. METHODS: Ninety-seven E. coli isolates from the bacteriology laboratory of The Research Center for Veterinary Science were identified on MacConkey medium supplemented with cefotaxime. The isolates were verified for the existence of the blaCTX-M, blaSHV, and blaTEM genes using PCR. Antimicrobial susceptibility testing was conducted using antibiotic discs following the CLSI standards. RESULTS: The prevalence of ESBL-producing E. coli in chicken ceca, eggs, and fish intestines was 16.5% (16/97). The specific genes detected were blaCTX-M gene at 93.75% (15/16), followed by the blaTEM gene, at 81.25% (13/16), and blaSHV at 25% (4/16). The antimicrobial sensitivity test results revealed that all ESBL-producing E. coli isolates had multidrug resistance 81.25% to 1-5 antibiotics and 18.75% to 6-7 antibiotics. The isolate exhibited 100% resistance to ampicillin and sulfamethoxazole, with exclusive sensitivity to chloramphenicol. CONCLUSION: The dominant gene in the ESBL-producing isolates was blaCTX-M. This bacterium is completely resistant to ampicillin and sulfamethoxazole, whereas it displays multidrug resistance to 1-7 different types of antibiotics.202540276175
146580.9991Detection of TEM, SHV and CTX-M in Mymensingh region in Bangladesh. The development of antibiotic resistance in bacteria following introduction of antimicrobial agents has emerged as an important medical problem everywhere in the world including Bangladesh. Extended spectrum β-lactamases (ESBLs) are rapidly evolving group of β-lactamase enzymes produced by the Gram negative bacteria. This study was undertaken to characterize ESBL producing gram negative bacilli from urine, skin wound (pus and wound infection). A total of 300 gram negative bacilli were screened for resistance to third generation Cephalosporins (3GCs) by disc diffusion test. The ESBL status was confirmed by double disc diffusion test (DDDT), minimum inhibitory concentration (MIC) by agar dilution method as recommended by Clinical Laboratory Standard Institute 2010 (CLSI) and multiplex PCR for TEM, SHV and CTX-M, CTX-M-3, CTX-M-14 genes. The present study revealed a higher occurrence of multi drugs resistant ESBLs production among gram negative isolates where Klebsiella spp. were the leading bacteria 36/45 (80%), followed by Proteus spp. 40/55 (72.7%), Esch. coli 105/156 (67.3%) and others 25/35 (71.4 %). Rate of TEM, SHV and CTX-M genes present in study population were 50.46%, 18.69% and 46.72% respectively. Among the CTX-M positive genes CTX-M-3 and CTX-M-14 were 78.0% (39/50) and 80.0% (40/50) respectively. Results indicate that routine ESBL detection should be made mandatory and irrational use of third generation cephalosporins must be discouraged to reduce multi drugs resistance bacteria, to increase patients' compliance and to make an antibiotic policy.201323982534
105790.9991Emergence of ciprofloxacin-resistant extended-spectrum β-lactamase-producing enteric bacteria in hospital wastewater and clinical sources. This study aimed to evaluate the incidence of ciprofloxacin-resistant extended-spectrum β-lactamase (ESBL)-producing enteric bacteria in hospital wastewater and clinical sources. Enteric bacteria, mainly Escherichia coli, were isolated from clinical sources (urinary tract and gastrointestinal tract infections; 80 isolates) and hospital wastewater (103 isolates). The antibiotic resistance profile and ESBL production of the isolates were investigated by disc diffusion assay and combined disc diffusion test, respectively. Plasmid profiling was performed by agarose gel electrophoresis, and elimination of resistance markers was performed by a plasmid curing experiment. Antibiotic susceptibility testing revealed a high incidence of β-lactam resistance, being highest to ampicillin (88.0%) followed by amoxicillin, ceftriaxone, cefpodoxime, cefotaxime, aztreonam, cefepime and ceftazidime. Among the non-β-lactam antibiotics, the highest resistance was recorded to nalidixic acid (85.7%). Moreover, 50.8% of enteric bacteria showed resistance to ciprofloxacin. Among 183 total enteric bacteria, 150 (82.0%) exhibited multidrug resistance. ESBL production was detected in 78 isolates (42.6%). A significantly higher incidence of ciprofloxacin resistance was observed among ESBL-producing enteric bacteria both in clinical (P=0.0015) and environmental isolates (P=0.012), clearly demonstrating a close association between ESBL production and ciprofloxacin resistance. Plasmid profiling of selected ESBL-positive strains indicated the presence of one or more plasmids of varying sizes. Plasmid curing resulted in loss of ciprofloxacin and cefotaxime resistance markers simultaneously from selected ESBL-positive isolates, indicating the close relationship of these markers. This study revealed a common occurrence of ciprofloxacin-resistant ESBL-producing enteric bacteria both in hospital wastewater and clinical sources, indicating a potential public health threat.201627436461
1464100.9991Detection of TEM and CTX-M genes from ciprofloxacin resistant Proteus mirabilis and Escherichia coli isolated on urinary tract infections (UTIs). The multidrug resistant Gram negative bacteria (MDRGNB) is an emerging burden and now represents a daily challenge for the management of antimicrobial therapy in healthcare settings. The present study was aimed to detect the prevalence of TEM and CTX-M type genes from GNB on urinary tract infection (UTIs). The ciprofloxacin resistant uropathogens were detected by HEXA UTI 5 disc diffusion method. The phenotypic detection of uropathogens producing extended spectrum beta lactamases (ESBLs) was confirmed by double disc combination test (DDCT) and phenotype confirmation test (PCT). The prevalence of TEM and CTX-M genes of uropathogens was identified by multiplex PCR analysis. The in vitro antimicrobial susceptibility of E. coli producing ESBL (26), 21 isolates of P. mirabilis, 17 P. aeruginosa, 14 K. pneumoniae and 6 Enterobacter sp. were detected. Based on the extension of the cephalosporin zone edge towards augmentin disc in the DDST method proved 84% of the isolates were ESBL positive. Similar results were obtained in phenotypic confirmatory test (PCT) by the increases of ≥5 mm zone of inhibition in the combination disc when compared with ceftazidime disc alone. The prevalence of TEM and CTX-M genes were determined from multidrug resistance uropathogens (MDU) respectively as 83%, 75%, 71%, 63%, 60%, 55%, 54%, 50%. The most prevalent (TEM + CTX-M) genes were also detected in ciprofloxacin resistant strains P. mirabilis BDUMS1 (KY617768) and E. coli BDUMS3 (KY617770). Due to the increase of ESBL genes in uropathogens, sustained supervision for using favorable antibiotics and decreasing the infection is essential.201829778819
1046110.9991Extended-spectrum β-lactamases producing multidrug resistance Escherichia coli, Salmonella and Klebsiella pneumoniae in pig population of Assam and Meghalaya, India. AIM: The present study was conducted to record the prevalence of extended spectrum β-lactamases (ESBLs) producing Escherichia coli, Salmonella spp., and Klebsiella pneumoniae from pig population of Assam and Meghalaya and to record the ability of the resistant bacteria to transfer the resistance genes horizontally. MATERIALS AND METHODS: Fecal samples (n=228), collected from pigs of Assam (n=99) and Meghalaya (n=129), were processed for isolation and identification of E. coli and Salmonella spp. All the isolates were tested for ESBLs production by double disc synergy test (DDST) followed by screening for ESBLs producing genes (bla(TEM), bla(SHV), bla(CTX-M), and bla(CMY)) by polymerase chain reaction (PCR). Possible transfer of resistance encoding genes between enteric bacterial species was carried out by in vitro and in vivo horizontal gene transfer (HGT) method. RESULTS: A total of 897 enteric bacteria (867 E. coli and 30 Salmonella) were isolated and identified. Altogether 25.41% isolates were confirmed as ESBL producers by DDST method. Majority of the isolates were E. coli followed by Salmonella. By PCR, 9.03% isolates were found positive for at least one of the target resistance genes. bla(SHV) was absent in all the isolates. bla(CMY) was the most prevalent gene. All the E. coli isolates from Assam were negative for bla(TEM). A total of 2.76% isolates were positive for bla(TEM) + bla(CMY). On the other hand, 0.67% isolates were positive for bla(CTX-M) + bla(CMY) genes. Only 0.33% isolates carried all the three genes. Altogether, 4.68% bacteria carried the resistance encoding genes in their plasmids. bla(TEM) gene could be successfully transferred from Salmonella (donor) to E. coli (recipient) by in vitro (5.5-5.7×10(-5)) and in vivo (6.5×10(-5) to 8.8×10(-4)) methods. In vivo method was more effective than in vitro in the transfer of resistance genes. CONCLUSION: The pig population of Assam and Meghalaya are carrying multidrug resistance and ESBLs producing E. coli and Salmonella. The isolates are also capable to transfer their resistance trait to other bacterial species by HGT. The present finding could be considered as a serious public health concern as similar trait can also be transmitted to the human commensal bacteria as well as pathogens.201830034183
991120.9991Characterization of extended-spectrum beta-lactamases in Enterobacteriaceae causing nosocomial infections in a Zagreb University Hospital. The bacteria producing extended-spectrum beta-lactamases (ESBLs) are increasingly reported. production of ESBLs by Gram-negative bacteria is the major mechanism of resistance to oxymino-cephalosporins and aztreonam. the aim of the present study was to characterize ESBLs produced by Enterobacteriaceae, collected during 2003-2005 in a University Hospital in Zagreb, and to determine the risk factors associated with nosocomial infections due to them. 76 isolates of Enterobacteriaceae were included in the study. Antibiotic susceptibility testing was performed by disk-diffusion and broth microdilution method according to CLSI. beta-lactamases were characterized by PCR and sequencing of bla(ESBL )genes. plasmids were extracted by alkaline lysis method and digested with EcoRI enzyme. Most of the strains displayed CAZ phenotype meaning a higher level of resistance to ceftazidime compared to cefotaxime and ceftriaxone. 50 strains produced SHV-ESBL, 28 tem and 8 CTX-M beta-lactamase. Sequencing of bla(SHV )genes from representative strains revealed SHV-5 beta-lactamase in 6 strains whereas sequencing of bla(CTX-M )genes identified CTX-M-3 beta-lactamase in 3 and CTX-M-15 in 5 strains. Strains were assigned to groups from A to f according to plasmid fingerprinting. The spread of SHV-5-producing strains throughout the hospital units could be due to selective pressure of ceftazidime which is widely prescribed in our hospital thus favoring survival of strains possessing a mutation at the Ambler position 240 responsible for ceftazidime and aztreonam resistance.200919567348
1124130.9991Molecular Identification of Extended-Spectrum β-lactamase and Integron Genes in Klebsiella Pneumonia. INTRODUCTION: Infections caused by Gram negative bacteria, producing extended-spectrum β-lactamase, including Klebsiella pneumoniae are increasing all over the world with high morbidity and mortality. The aim of the present study was determined antimicrobial profile susceptibility and the prevalence of antibiotic resistance genes by multiplex PCR. METHODS: In the present study, we obtained one-hundred isolates of K. pneumoniae from different clinical samples. The antibiotic susceptibility testing was done in thirteen antibiotic and, therefore, M-PCRs were conducted using the DNA amplification for detection of ESBLs (blaTEM, blaCTX-M, blaSHV) and int (I, II, III) genes. RESULTS: The results of resistance to amoxicillin/clavulanate, ciprofloxacin, amikacin, trimethoprim-sulfamethoxazole, cefotaxime, ampicillin, aztreonam, imipenem, gentamicin, ceftazidime, Cefepime, ceftriaxone and levofloxacin were obtained 37%, 37%, 93%, 84%, 52%, 87%, 59%, 8%, 24%, 67%, 52%, 43% and 26%, respectively. The frequency of the extended-spectrum β-lactamase K. pneumoniae was obtained 37%. The prevalence of resistance genes of ESBLs in the M-PCR method showed that the blaTEM, blaCTX and blaSHV were 38%, 24% and 19%, respectively, however, only 8 (8%) out of 100 isolates were found to have positive outcomes for the existence of class 1 integrons and there were no detected class 2 or class 3 integrons. CONCLUSIONS: Our results recommend the likely co-carriage of some ESBLs genes and antibiotic resistance integrons on the same plasmids harboring the MDR genes.201627935927
1166140.9991Yearly incidence of acute childhood gastroenteritis in Nigeria: Implicated pathogens predominantly harbor blaCTXM and blaTEM genes. BACKGROUND AND OBJECTIVES: Routine use of antibiotics for infectious diarrhea in children is associated with the risk of increasing antibiotic resistance in developing countries. This work aimed to study the predominant extended spectrum beta-lactamase (ESBL) genes among bacteria pathogens implicated in acute childhood gastroenteritis in a tertiary hospital in Nigeria. MATERIALS AND METHODS: The stool samples of children diagnosed with acute gastroenteritis were collected. Isolation and identification of bacterial pathogens from the stool samples using standard microbiological and molecular sequencing methods. Pure cultures of the probable bacteria pathogens were subjected to antibiotics susceptibility profiling using the Kirby-Bauer Disk Diffusion Method and also screened for ESBL and AmpC using the Modified Double Disc Synergy Test. Primers for 5 different ESBL genes associated with beta-lactam antibiotic resistance were amplified and sequenced. RESULTS: Out of the 62 isolates, the highest number of organisms identified within the isolates were Bacillus sp at 38.7% (24) followed by Alcaligenes sp at 37% (23). Resistance to cefepime and ceftazidime were recorded at 50.8% (30) each. Ceftriaxone and cefotaxime were resisted in 47.4% (28) of the isolates. Out of 34 isolates resistant to all the cephalosporins used, 41.2% (14) were ESBL-producing, of which blaCTXM-1 and blaCTXM-2 were detected in 85.7%, while blaTEM was seen in 64.3%. CONCLUSIONS: blaCTXM and blaTEM may be the predominant ESBL genes haboured in the bacteria pathogens implicated in the yearly incidence of acute childhood gastroenteritis in Nigeria. This may be due to the widespread use of antibiotics in treating this disease.202539977466
1127150.9991Extended spectrum beta-lactamase and aminoglycoside modifying enzyme genes in multi drug resistant Gram-negative bacteria: A snapshot from a tertiary care centre. BACKGROUND: This study aims to enhance the existing knowledge of the prevalence of genes responsible for beta-lactam resistance and aminoglycoside resistance in gram negative organisms by molecular detection of extended spectrum beta-lactamase and aminoglycoside modifying enzymes in multidrug-resistant gram-negative bacteria. METHODS: Out of 864 gram-negative isolates, 710 were phenotypically identified as multidrug-resistant by antibiotic susceptibility testing. From the above isolates, 102 representative isolates as per sample size calculated were selected for further molecular studies. The presence of blaTEM, blaCTX-M blaSHV, and five AmpC genes was detected by real-time polymerase chain reaction (PCR). Conventional PCR was performed to detect seven aminoglycoside modifying enzyme genes namely aac(6')-Ib, aac(6')-Ic, aac(3)-Ia, aac(3)-Ib, aac(3)-IIa, ant(2'')-Ia, and ant(4'')-IIa. RESULTS: Most common multidrug-resistant isolate was Klebsiella pneumoniae (35%) followed by Escherichia coli (30%). Among the 102 selected isolates all harboured blaTEM gene, 71 (69.6%) harboured blaCTX-M gene and 48 (47%) blaSHV gene. Among the selected isolates 60% showed the presence of AmpC genes. Most common aminoglycosie modifying enzyme gene was AAC 6' Ib (51%) followed by ANT 2" Ia (36%). CONCLUSION: This study suggests a wider use of molecular methods using specific PCR amplification of resistance genes. It would be beneficial to perform the molecular identification of antimicrobial resistance genes to effectively monitor and manage antibiotic resistance, administer appropriate antimicrobial medication, practice antimicrobial stewardship and improve hospital infection control procedures.202439734850
1054160.9991Molecular detection of extended-spectrum β-lactamase-producing Klebsiella pneumoniae isolates of chicken origin from East Java, Indonesia. BACKGROUND AND AIM: Klebsiella pneumoniae is one of the respiratory disease agents in human and chicken. This bacterium is treated by antibiotic, but this treatment may trigger antibiotic resistance. Resistance gene in K. pneumoniae may be transferred to other bacteria. One of the known resistance genes is extended-spectrum β-lactamase (ESBL). This research aimed to study K. pneumoniae isolated from chicken farms in East Java, Indonesia, by observing the antibiotic resistance pattern and detect the presence of ESBL coding gene within the isolates. MATERIALS AND METHODS: A total of 11 K. pneumoniae isolates were collected from 141 chicken cloacal swabs from two regencies in East Java. All isolates were identified using the polymerase chain reaction method. Antimicrobial susceptibility was determined by agar dilution method on identified isolates, which then processed for molecular characterization to detect ESBL coding gene within the K. pneumoniae isolates found. RESULTS: The result of antibiotic sensitivity test in 11 isolates showed highest antibiotic resistance level toward ampicillin, amoxicillin, and oxytetracycline (100%, 100%, and 90.9%) and still sensitive to gentamicin. Resistance against colistin, doxycycline, ciprofloxacin, and enrofloxacin is varied by 90.9%, 54.5%, 27.3%, and 18.2%, respectively. All isolates of K. pneumoniae were classified as multidrug resistance (MDR) bacteria. Resistance gene analysis revealed the isolates harbored as bla (SHV) (9.1%), bla (TEM) (100%), and bla (CTX-M) (90.9%). CONCLUSION: All the bacterial isolates were classified as MDR bacteria and harbored two of the transmissible ESBL genes. The presence of antibiotic resistance genes in bacteria has the potential to spread its resistance properties.201931190714
1123170.9991Molecular detection of blaSHV gene in multidrug resistance of Klebsiella pneumoniae isolated from chicken egg shell swab from a traditional market in Surabaya. BACKGROUND: Contamination with Klebsiella pneumoniae in food ingredients, including eggs, causes various dangers because it threatens public health, because it acts as a multidrug resistance (MDR) bacteria, especially the extended-spectrum beta-lactamase (ESBL) strain. The ESBL blaSHV gene is part of a broad-spectrum ESBL that is often found in Gram-negative bacteria. AIM: This study aimed to identify the ESBL blaSHV gene in K. pneumoniae MDR from chicken eggshells. METHODS: This study used 160 samples of chicken eggshell swabs isolated on 1% BPW media from 10 traditional Surabaya markets. Samples were isolated using MCA media and were identified using Gram staining and biochemical tests. Detection of MDR using Muller-Hinton Agar. RESULTS: Confirmation of ESBL in multidrug-resistant (MDR) isolates was performed using polymerase chain reaction to detect ESBL genes. The results showed that the isolation and identification of K. pneumoniae bacteria were 25.62% (41/160). Amoxicillin antibiotics showed the highest level of resistance at a percentage of 100% (41/41), followed by antibiotic resistance to erythromycin (90.24% (37/41), Streptomycin antibiotics were 26.82% (11/41), ciprofloxacin (14.63% (6/41), and Tetracycline antibiotic resistance was 7.31% (3/41). The results of MDR from K. pneumoniae showed 34.14% (14/41) of the isolates were then tested by PCR, which showed positive results for the blaSHV gene of 71.42% (10/14). CONCLUSION: The data from this study confirm the existence of K. pneumoniae bacteria isolated from egg shell swabs carrying the blaSHV gene from MDR isolates.202540557075
1165180.9991Isolation, Antimicrobial Susceptibility Profile and Detection of Sul1, blaTEM, and blaSHV in Amoxicillin-Clavulanate-Resistant Bacteria Isolated From Retail Sausages in Kampar, Malaysia. BACKGROUND: Due to the overuse of antibiotics in livestock as a growth-promoting agent, the emergence of multi-antibiotic resistant bacteria is becoming a concern. OBJECTIVES: In this study, we aimed to detect the presence and discover the molecular determinants of foodborne bacteria in retail sausages resistant towards the antibacterial agent amoxicillin-clavulanate. METHODS: Two grams of sausages were chopped into small pieces and transferred into sterile Luria-Bertani (LB) enrichment broths overnight before they were plated on MacConkey agar petri dishes. The bacteria isolated were then screened for amoxicillin-clavulanate resistance, and an antimicrobial susceptibility test of each isolate was performed by using the disc diffusion method. Double synergy and phenotypic tests were carried out to detect the presence of extended spectrum β-lactamase (ESBL). API 20E kit was used to identify the Enterobacteriaceae. All isolates were further examined by polymerase chain reaction (PCR) for resistant genes blaOXA-1, blaOXA-10, plasmid-mediated AmpC (blaCMY and blaDHA), and the chromosome-mediated AmpC, Sul1, blaTEM, and blaSHV genes. RESULTS: A total of 18 amoxicillin-clavulanate resistant isolates were obtained from seven different types of retail sausages. Only half of them were identified as Enterobacteriaceae, but none were ESBL-producers. All the 18 isolated strains demonstrated resistance towards amoxicillin-clavulanate, penicillin and oxacillin (100%), cefotaxime (71.4%), cefpodoxime (66.7%), and ampicillin (83.3%). blaTEM was the most frequently detected β-lactamase gene. Both plasmid- and chromosomal-bound blaTEM genes were detected in all of the isolated Enterobacteriaceae. blaSHV and Sul1 accounted for 22.2% and 11.1% of the amoxicillin-clavulanate resistant isolates, respectively, whereas blaAMPC, blaCMY, blaDHA, blaOXA-1, and blaOXA-10 were not found in any of the isolates. The only one ESBL-producing bacteria detected in this study was Chryseobacterium meningosepticum, which harbored the blaTEM gene. CONCLUSIONS: The multidrug resistant bacteria that carry antibiotic resistant genes from retail sausages may increase the risk of transmission to humans via the consumption of contaminated sausages. Stricter measures must be taken to address the use of antibiotics in animal agriculture and to consider their potential impact on human health.201627942365
1061190.9991Genotypic characterization of bacterial isolates causing urinary tract infections among adults at Kiambu Level 5 Hospital, Kenya: selected extended-spectrum β-lactamase genes and biofilm formation. The menace of antimicrobial resistance affecting public health is rising globally. Many pathogenic bacteria use mechanisms such as mutations and biofilm formation, significantly reducing the efficacy of antimicrobial agents. In this cross-sectional study, we aimed to determine the prevalence of selected extended-spectrum β-lactamase (ESβL) genes and analyse the biofilm formation abilities of the isolated bacteria causing urinary tract infection among adult patients seeking Medicare at Kiambu Level 5 Hospital, Kenya. The double-disc synergy test was used for phenotypic identification of ESβL-producing isolates, while microtitre plate assays with some modifications were used for the biofilm formation test. Ten isolates were bioassayed for ESβL genes out of 57 bacterial isolates obtained from urine samples. This study found the bla (TEM) genes to be the most prevalent ESβL type [10/10 (100 %)], followed by bla(OXA) and bla(SHV) genes at 4/10 (40 %) and 3/10 (30 %), respectively. In addition, co-carriage of bla(TEM) and bla(SHV) was 50 % lower than that of bla(TEM)+bla (OXA) genes at 66.7 % among Escherichia coli isolates studied. Biofilm formation was positive in 36/57 (63.2 %) of the isolates tested, with most being Gram-negative [25/36 (69.4 %)]. Escherichia coli [15/36 (41.7 %)], Klebsiella species [7/36 (19.4 %)] and Staphylococcus aureus [7/36 (19.4 %)] were the dominant biofilm formers. However, there was no significant difference in biofilm formation among all tested isolates, with all isolates recording P-values >0.05. In light of these findings, biofilm formation potential coupled with antimicrobial resistance genes in urinary tract infection isolates may lead to difficult-to-treat infections.202438482364