Molecular Identification of Extended-Spectrum β-lactamase and Integron Genes in Klebsiella Pneumonia. - Related Documents




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112401.0000Molecular 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
112310.9999Molecular 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
112720.9999Extended 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
98630.9999The Frequency of qnr Genes in Extended-Spectrum β-lactamases and non-ESBLs Klebsiella pneumoniae Species Isolated from Patients in Mashhad, Iran. BACKGROUND AND OBJECTIVES: Since the fluoroquinolones are the broad-spectrum antibiotics, they affect both Gram-negative and Gram-positive bacteria. These antibiotics are widely prescribed by physicians. As a result, some bacteria, especially Enterobacteriaceae, have shown a resistance to this family of antibiotics. The current study aimed at detecting the frequency of qnrA, qnrB, and qnrS genes, novel plasmid-mediated quinolone-resistance genes, among extended-spectrum β-lactamases (ESBL)-positive and ESBL-negative Klebsiella pneumoniae isolates. MATERIALS AND METHODS: One hundred and thirty isolates of K. pneumoniae were collected from Imam Reza Hospital and its associated clinics from May 2011 to July 2012. The isolates were tested for ESBLs by the conventional methods. Polymerase chain reaction (PCR) was performed to amplify qnr A, B, and S. RESULTS: Thirty-eight (29.3%) isolates were ciprofloxacin-resistant. Among 130 K. pneumoniae infectious isolates, 56 (43%) were capable of producing ESBL; 10.8% (n=14), 15.4% (n=20), and 20.8% (n=27) of ESBL-producing K. pneumonia were positive for qnrA, qnrS, and qnrB, respectively, and 13.8% (n=18) of the isolates harbored 2 or 3 qnr genes. CONCLUSION: The results of the current study showed that quinolone-resistance genes were more frequent in ESBL-producing K. pneumoniae (37.5%) isolates, compared with the ESBL-negative isolates (20.89%). The prevalence of qnr genes was high in K. pneumoniae isolates, with higher frequency in ESBL-positive strains. Most of the isolates were positive for all 3 groups of qnr genes and the qnrB was the most common one.201729563934
111640.9999Detection of blaCTX-M genes in ESBL-producing Klebsiella isolates from animals in Croatia. This study investigated the frequency of third-generation cephalosporin resistance in Klebsiella spp. isolated from animals in Croatia and the presence of blaCTX-M genes. A total of 711 enteric bacteria were isolated from clinical samples, and Klebsiella spp. isolates accounted for 6.9% (n = 49). Thirteen Klebsiella isolates (26.5%) were ESBL producers, nine isolates from the Klebsiella pneumoniae species complex (69.2%), and four (30.8%) Klebsiella oxytoca isolates. All carried the blaCTX-M-15 gene, and antimicrobial susceptibility testing revealed them as multidrug resistant. All were resistant to all tested cephalosporins, fluoroquinolones, aminoglycosides and aztreonam, 92.3% showed resistance to tetracycline, 84.6% to trimethoprim-sulfamethoxazole and 69.2% to nitrofurantoin. No isolate showed resistance to imipenem and meropenem. It can be concluded that ESBL-producing Klebsiella expressing the blaCTX-M gene are not rare among Klebsiella isolates from animals in Croatia.202337027245
144750.9999Molecular detection of β-lactamase and integron genes in clinical strains of Klebsiella pneumoniae by multiplex polymerase chain reaction. INTRODUCTION: Infections caused by β-lactamase-producing gram-negative bacteria, such as Klebsiella pneumoniae, are increasing globally with high morbidity and mortality. The aim of the current study was to determine antimicrobial susceptibility patterns and the prevalence of antibiotic resistance genes (β-lactamase and integron genes) using multiplex PCR. METHODS: One-hundred K. pneumoniae isolates were collected from different clinical samples. Antibiotic susceptibility testing was performed with thirteen different antibiotics. Multiplex-PCR was used to detect β-lactamase (bla TEM, bla CTX-M, bla SHV , bla VEB, bla PER, bla GES, bla VIM, bla IMP, bla OXA, and bla KPC) and integron genes (int I, int II, and int III). RESULTS: The highest and lowest rate of resistance was exhibited against amikacin (93%) and imipenem (8%), respectively. The frequency of β-lactamase-positive K. pneumoniae was 37%, and the prevalence of the bla TEM, bla CTX-M, bla SHV , bla VEB, bla PER, bla GES, bla VIM, bla IMP, bla OXA, and bla KPC genes was 38%, 24%, 19%, 12%, 6%, 11%, 33%, 0%, 28%, and 23%, respectively. Of the 100 isolates, eight (8%) were positive for class I integrons; however, class II and III integrons were not detected in any of the strains. CONCLUSIONS: These results indicate co-carriage of a number of β-lactamase genes and antibiotic resistance integrons on the same plasmids harboring multi-drug resistance genes. It seems that these properties help to decrease treatment complications due to resistant bacterial infections by rapid detection, infection-control programs and prevention of transmission of drug resistance.201728700049
106960.9999High Prevalence of Antimicrobial Resistance in Gram-Negative Bacteria Isolated from Clinical Settings in Egypt: Recalling for Judicious Use of Conventional Antimicrobials in Developing Nations. This study was designed to investigate, at the molecular level, the antimicrobial resistance mechanisms of different antimicrobial resistance genes, including, extended-spectrum β-lactamases, AmpC β-lactamases, class 1 and 2 integrons, and plasmid-mediated quinolone resistance genes of Gram-negative bacteria isolated from clinical settings in Egypt. A total of 126 nonduplicate Gram-negative isolates were recovered from different clinical samples taken from hospitalized patients in Egypt in 2014. Antimicrobial susceptibility testing showed that, 93.6% (118/126) of the isolates had a multidrug-resistant phenotype. Interestingly, we reported a high level of antimicrobial resistance nearly for all tested antibiotics; to our knowledge, this is the first report from Egypt indicating very high level of antibiotic resistance in Egypt. Polymerase chain reaction screening and DNA sequencing revealed that, 75.4% (95/126) of the isolates harbored at least one extended-spectrum β-lactamase-encoding gene, with bla(CTX-M) being the most prevalent (65.9%), followed by bla(SHV) (46.8%). The AmpC β-lactamase, bla(CMY), was detected in 7.1% (9/126) of bacterial isolates, with bla(CMY-42) being the most prevalent. Class 1 integrons were detected in 50.8% (64/126) of the isolates, and class 2 integrons were detected in 2.4% (3/126) of the isolates. The plasmid-mediated quinolone resistance gene, qnr, was detected in 58.7% (74/126) of the tested isolates, with qnrS being the most prevalent. Several antimicrobial resistance determinants were identified in Egypt for the first time, such as SHV-27, SHV-28, SHV-33, SHV-63, SHV-71, SHV-82, SHV-142, CMY-42, CMY-6, and the new CMY-72 like. This study highlights the importance of the conscious use of conventional antimicrobials to overcome the multidrug resistance problem.201930681401
112870.9999Molecular 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
105480.9999Molecular 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
98790.9999Characterization 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
1125100.9999Detection of emerging antibiotic resistance in bacteria isolated from subclinical mastitis in cattle in West Bengal. AIM: The aim of this work was to detect antibiotic resistance in Gram-negative bacteria isolated from subclinical mastitis in cattle in West Bengal. MATERIALS AND METHODS: The milk samples were collected from the cattle suffering with subclinical mastitis in West Bengal. The milk samples were inoculated into the nutrient broth and incubated at 37°C. On the next day, the growth was transferred into nutrient agar and MacConkey agar. All the pure cultures obtained from nutrient agar slant were subjected to Gram-staining and standard biochemical tests. All the bacterial isolates were tested in vitro for their sensitivity to different antibiotics commonly used in veterinary practices. All Gram-negative isolates including positive control were subjected to polymerase chain reaction (PCR) for detection of bla(CTX-M), bla(TEM), bla(SHV), bla(VIM), tetA, tetB, tetC, and tetM genes considered for extended-spectrum β-lactamase (ESBL), metallo-β-lactamase, and tetracycline resistance. RESULTS: In total, 50 Gram-negative organisms (Escherichia coli, Proteus, Pseudomonas, Klebsiella, and Enterobacter) were isolated from milk samples of subclinical mastitis infected cattle. Among these Gram-negative isolates, 48% (24/50) were found either ESBL producing or tetracycline resistant. Out of total 50 Gram-negative isolates, bla(CTX-M) was detected in 18 (36%) isolates, and 6 (12%) harbored bla(TEM) genes in PCR. None of the isolates carried bla(SHV) genes. Further, in this study, 5 (10%) isolates harbored tet(A) gene, and 8 (16%) isolates carried tet(B) gene. No tet(C) gene was detected from the isolates. CONCLUSION: This study showed emerging trend of antibiotic-resistant Gram-negative bacteria associated with subclinical mastitis in cattle in West Bengal, India.201728620255
1126110.9999Comparison of beta-lactamase genes in clinical and food bacterial isolates in India. BACKGROUND: The present study aimed to determine the occurrence of human disease-causing enteric bacteria on raw vegetables, fruits, meats, and milk products sold in Indian markets. The study further aimed to analyze antibiotic resistance rates and the presence of blaCTX-M, blaTEM, blaSHV, and blaAmpC. METHODOLOGY: Twenty-three food-borne and 23 clinical isolates were compared for antibiotic resistance rates and the presence of blaCTX-M, blaTEM, blaSHV, and blaAmpC. Swabs were taken from unwashed and washed food items, as well as from some chopped food specimens, and inoculated on appropriate culture medium. Bacterial isolates were identified, antibiotic susceptibility was performed, and bla genes were detected by PCR. RESULTS: Thirty-eight bacterial isolates were obtained from the food specimens, of which 36 (94.7%) were Gram-negative and two (5.3%) were Gram-positive bacterial species. Klebsiella pneumoniae was the most prevalent (52.6%; 20/38) bacterial species isolated, followed by Citrobacter koseri (18.4%; 7/38). In food isolates, the majority of the isolates were resistant to gentamicin (33.3%) followed by amikacin (11.1%). Resistance to a third-generation cephalosporin was noticed in only 5.6% isolates. However, in clinical isolates, maximal resistance was noticed against third-generation cephalosporins followed by ofloxacin in 91.3% and 86.9% isolates, respectively, and resistance to gentamicin and amikacin was noticed in 78.3% and 52.2% isolates, respectively. The presence of blaCTX-M, blaTEM, blaSHV, and blaAmpC in clinical isolates was noticed in 52.2%, 60.9%, 21.7%, and 43.5%, respectively. None of the isolates from food showed the presence of any of the above-cited genes. CONCLUSIONS: Probably bla genes have not yet disseminated to raw-food vegetation in India.200919801801
2111120.9999Antimicrobial Resistance and Resistance Determinant Insights into Multi-Drug Resistant Gram-Negative Bacteria Isolates from Paediatric Patients in China. INTRODUCTION: The emergence of multi-drug-resistant Gram-negative bacteria (GNB) is a concern in China and globally. This study investigated antimicrobial resistance traits and resistance determinant detection in GNB isolates from paediatric patients in China. METHODS: In the present study, a total of 170 isolates of GNB including the most prevalent Escherichia coli, Klebsiella pneumoniae and Acinetobacter baumannii were collected from Shenzhen Children's Hospital, China. ESBLs production was confirmed by using the combination disc diffusion method, and carbapenemase production was confirmed by using a carbapenem inactivation method followed by antimicrobial susceptibility. In addition, β-lactamase-encoding genes and co-existence of plasmid-borne colistin resistance mcr-1 gene were determined by PCR and sequencing. RESULTS: Overall, 170 etiological agents (GNB) were recovered from 158 paediatric patients. The most prevalent species was E. coli 40% (n=68), followed by K. pneumoniae 17.64% (n=30), and Enterobacter cloacae 14.11% (n=24). Of 170 GNB, 71.76% (n=122) were multi-drug-resistant, 12.35% (n=21) extreme-drug resistant, and 7.64% (n=13) single-drug-resistant, while 8.23% (n=14) were sensitive to all of the studied antibiotics. The prevalence of ESBLs and carbapenemase producers were 60% and 17%, respectively. bla (CTX-M) was the most prevalent resistance gene (59.42%), followed by bla (TEM) (41.17%), bla (SHV) (34.270%), bla (KPC) (34.11%), bla (OXA-48) (18.82%) and bla (NDM-1) (17.64%). CONCLUSION: The present study provides insights into the linkage between the resistance patterns of GNB to commonly used antibiotics and their uses in China. The findings are useful for understanding the genetics of resistance traits and difficulty in tackling of GNB in paediatric patients.201931819545
1446130.9999One-Day Prevalence of Extended-Spectrum β-Lactamase (ESBL) and Carbapenemase-Producing Bacteria in Fecal Samples from Surgical Patients: A Concerning Trend of Antibiotic Resistance. PURPOSE: Extended-spectrum β-lactamase (ESBL) and carbapenemase producing bacteria are of increasing concern due to their multidrug resistance and infection potential. This study determines the one-day prevalence of faecal carriage of ESBL and carbapenemase producing Gram-negative bacilli. METHODS: Fecal samples were collected from 30 post-surgery patients (hospitalized for at least 48 hours) in each of the four hospitals involved in the study and were analyzed for antibiotic-resistant bacteria. Identification was done using Maldi Tof mass spectrometry, and antibiotic susceptibility was tested using disk diffusion and specialized tests for ESBL (double disk synergy technique) and carbapenem (NG-TEST CARBA 5) resistance detection. PCR was conducted on isolates to detect betalactam resistance genes, carbapenemase genes and quinolone resistance genes. FINDINGS: Out of the 120 patients enrolled, 38.33% (n = 46) and 49.16.33% (n = 59) were found to carry ESBL- and carbapenemase-producing bacteria, respectively, in their fecal samples. Among the isolates, 51.08% (n = 47) exhibited ESBL production, with Escherichia coli (44.56%) being the most common species. The identification of bacteria with resistance to carbapenems showed a predominance of the species Escherichia coli (44.45%) followed by the species Klebsiella pneumoniae (16.06%) and Acinetobacter baumanii (13.58%). The study of the association of variables shows a high degree of association (p < 0.05) for the factors independent walking and use of a wheelchair with ESBL production. The most frequently detected genes among ESBL producing bacteria were bla(CTXM-1) (91.49%), qnrB (70.21%) and qnrs (63.82%). bla(NDM) (54.68%) was the most detected carbapenemase genes among carbapenemase producing isolates. CONCLUSION: This study demonstrates, for the first time, a significant prevalence of ESBL and carbapenemase producing gram-negative bacteria among surgical patients in Benin, with multiple resistance genes detected. Findings should be interpreted in light of the cross-sectional design and >48-hour hospitalization criterion.202540635768
1115140.9999Prevalence of extended spectrum beta lactamase and plasmid mediated quinolone resistant genes in strains of Klebsiella pneumonia, Morganella morganii, Leclercia adecarboxylata and Citrobacter freundii isolated from poultry in South Western Nigeria. A serious concern is arising on the coexistence of extended-spectrum beta-lactamase (ESBL) and plasmid mediated quinolone resistance (PMQR) producing bacteria in animal husbandry, which could be transferred to humans, especially in strains that may not be routinely screened for resistance. This study therefore tested the prevalence of ESBL and PMQR genes in selected bacteria isolated from poultry faeces. Faecal droppings of birds were collected from 11 farms in five states in South Western Nigeria. Bacteria were isolated from the samples on cefotaxime supplemented plates and identified with MALDI-TOF. The MIC was determined using VITEK system and resistance genes were detected with PCR. A total of 350 strains were isolated from different samples and selected strains were identified as 23 Klebsiella pneumonia, 12 Morganella morganii, seven Leclercia adecarboxylata and one Citrobacter freundii. All the species were resistant to gentamycin, trimethoprim/sulphamethaxole, tobramycin, piperacillin, cefotaxime and aztreonam (except Morganella morganii strains which were mostly susceptible to aztreonam). All the tested strains were susceptible to imipenem, meropenem and amikacin. All Leclercia adecarboxylata strains were resistant to ceftazidime, cefepime and fosfomycin while all Morganella morganii strains were resistant to fosfomycin, moxifloxacin and ciprofloxacin. All tested species were generally sensitive to ciprofloxacin except Morganella morganii strains which were resistant to ciprofloxacin. The resistance to ciprofloxacin, ceftazidime, cefepime, tigercylin, colistin and fosfomycin were 65%, 40%, 23%,, 7%, 33%, 48% respectively while the prevalence of SHV, TEM and CTX genes were 42%, 63%, 35% respectively. 9.3% of the isolates had the three ESBL genes, 2.33% had qnrA gene, 4.65% had qnr B gene while none had qnrS gene. The most prevalent PMQR gene is Oqxb (25.58%) while 6.98% had the qep gene. Klebsiella pneumoniae generally had both ESBL and PMQR genes. The high prevalence of extended spectrum beta-lactamase genes in the studied strains calls for caution in the use of beta lactam antibiotics in poultry feeds. This is the first report of the occurrence of extended spectrum beta-lactamase and plasmid mediated quinolone resistance genes in Morganella morganii and Leclercia adecarboxylata strains isolated from poultry faeces.201829942700
945150.9999Extended Spectrum Beta Lactamase (ESBL), bla(TEM),bla(SHV) and bla(CTX-M), Resistance Genes in Community and Healthcare Associated Gram Negative Bacteria from Osun State, Nigeria. BACKGROUND: Extended Spectrum Beta Lactamase (ESBL) production in gram negative bacteria confers multiple antibiotic resistance, adversely affecting antimicrobial therapy in infected individuals. ESBLs result from mutations in β-lactamases encoded mainly by the bla(TEM),bla(SHV) and bla(CTX-M) genes. The prevalence of ESBL producing bacteria has been on the increase globally, especially its upsurge among isolates from community-acquired infections has been observed. AIM: To determine ESBL prevalence and identify ESBL genes among clinical isolates in Osun State, Nigeria. MATERIAL AND METHODS: A cross-sectional study was carried out from August 2016 - July 2017 in Osun State, Nigeria. Three hundred and sixty Gram-negative bacteria recovered from clinical samples obtained from both community and healthcare-associated infections were tested. They included 147 Escherichia coli (40.8%), 116 Klebsiella spp (32.2%), 44 Pseudomonas aeruginosa (12.2%) and 23 Proteus vulgaris (6.4%) isolates. Others were Acinetobacter baumannii, Serratia rubidae, Citrobacter spp, Enterobacter spp and Salmonella typhi. Disk diffusion antibiotic susceptibility testing was carried out, isolates were screened for ESBL production and confirmed using standard laboratory procedures. ESBLs resistance genes were identified by Polymerase Chain Reaction (PCR). RESULTS: All isolates demonstrated multiple antibiotic resistance. Resistance to ampicillin, amoxicillin with clavulanate and erythromycin was 100%, whereas resistance to Imipenem was very low (5.0%). The overall prevalence of ESBL producers was 41.4% with Klebsiella spp as the highest ESBL producing Enterobacteriacaea. ESBL producers were more prevalent among the hospital pathogens than community pathogens, 58% vs. 29.5% (p=0.003). ESBL genes were detected in all ESBL producers with the bla(CTX-M) gene predominating (47.0%) followed by bla(TEM) (30.9%) and bla(SHV) gene was the least, 22.1%. The bla(CTX-M) gene was also the most prevalent in the healthcare pathogens (62%) but it accounted for only 25% in those of community origin. CONCLUSION: A high prevalence of ESBL producing gram-negative organisms occurs both in healthcare and in the community in our environment with the CTX-M variant predominating. Efforts to control the spread of these pathogens should be addressed.202132729432
1118160.9999Detection and characterization of extended-spectrum β-lactamases (blaCTX-M-1 and blaSHV ) producing Escherichia coli, Salmonella spp. and Klebsiella pneumoniae isolated from humans in Mizoram. AIM: The present study was conducted to isolate and characterize the extended spectrum β-lactamases (ESBLs) producing enteric bacteria in human beings in Mizoram, India. MATERIALS AND METHODS: Fecal samples were collected from human beings with or without the history of diarrhea from different hospitals of Mizoram. Samples were processed for isolation and identification of Escherichia coli, Salmonella and Klebsiella pneumoniae. All the isolates were subjected to antibiotic sensitivity assays. Phenotypically, ESBLs production ability was determined by double discs synergy test (DDST) method. ESBLs producing isolates were subjected to polymerase chain reaction (PCR) for detection of ESBLs genes. Plasmids were cured by acridine orange. Transfer of resistance from a donor to recipient strains was done by in vitro horizontal method. RESULTS: A total of 414 enteric bacteria were isolated from 180 fecal samples (113 were from diarrheic patients and 67 were from non-diarrheic patients), of which 333 (80.44%), 52 (12.56%), and 29 (7.00%) were E. coli, K. pneumoniae and Salmonella spp., respectively. Double discs synergy test (DDST) exhibited 72 (21.62%) E. coli, 12 (23.08%) K. pneumoniae and 4 (13.79%) Salmonella spp. were ESBLs producers. Altogether, 24 (13.04%) isolates were found to be positive for at least one resistance genes under this study. A total of 36 (8.70%) E. coli, 4 (0.97%) K. pneumoniae and 2 (0.48%) Salmonella spp. were found to be positive for blaCTX-M-1 gene by PCR. Similarly, 5 (1.21%) E. coli and 4 (0.97%) K. pneumoniae isolates were found to be positive for blaSHV gene. A total of 3 (0.72%) K. pneumoniae isolates were recorded as positive for both blaCTX-M-1 and blaSHV genes. All the isolates were carrying plasmids ranging between 0.9 kb and ~30 kb. The resistance plasmid could not be transferred to a recipient by in vitro horizontal gene transfer method. CONCLUSION: ESBLs producing enteric bacteria are circulating in human population in North Eastern Region of India. Indiscriminate use of antibiotics should be avoided to control the menace of multidrug resistance bacteria in the environment, animals, and human beings.201527047141
2176170.9999Evaluation of phenotypic and genotypic patterns of aminoglycoside resistance in the Gram-negative bacteria isolates collected from pediatric and general hospitals. The purpose of the current study was to evaluate the phenotypic and genotypic patterns of aminoglycoside resistance among the Gram-negative bacteria (GNB) isolates collected from pediatric and general hospitals in Iran. A total of 836 clinical isolates of GNB were collected from pediatric and general hospitals from January 2018 to the end of December 2019. The identification of bacterial isolates was performed by conventional biochemical tests. Susceptibility to aminoglycosides was evaluated by the disk diffusion method (DDM). The frequency of genes encoding aminoglycoside-modifying enzymes (AMEs) was screened by the PCR method via specific primers. Among all pediatric and general hospitals, the predominant GNB isolates were Acinetobacter spp. (n = 327) and Escherichia coli (n = 144). However, E. coli (n = 20/144; 13.9%) had the highest frequency in clinical samples collected from pediatrics. The DDM results showed that 64.3% of all GNB were resistant to all of the tested aminoglycoside agents. Acinetobacter spp. and Klebsiella pneumoniae with 93.6%, Pseudomonas aeruginosa with 93.4%, and Enterobacter spp. with 86.5% exhibited very high levels of resistance to gentamicin. Amikacin was the most effective antibiotic against E. coli isolates. In total, the results showed that the aac (6')-Ib gene with 59% had the highest frequency among genes encoding AMEs in GNB. The frequency of the surveyed aminoglycoside-modifying enzyme genes among all GNB was found as follows: aph (3')-VIe (48.7%), aadA15 (38.6%), aph (3')-Ia (31.3%), aph (3')-II (14.4%), and aph (6) (2.6%). The obtained data demonstrated that the phenotypic and genotypic aminoglycoside resistance among GNB was quite high and it is possible that the resistance genes may frequently spread among clinical isolates of GNB.202235119565
1049180.9999Multiple Antibiotic-Resistant, Extended Spectrum-β-Lactamase (ESBL)-Producing Enterobacteria in Fresh Seafood. Members of the family Enterobacteriaceae include several human pathogens that can be acquired through contaminated food and water. In this study, the incidence of extended spectrum β-lactamase (ESBL)-producing enterobacteria was investigated in fresh seafood sold in retail markets. The ESBL-positive phenotype was detected in 169 (78.60%) isolates, with Escherichia coli being the predominant species (53), followed by Klebsiella oxytoca (27), and K. pneumoniae (23). More than 90% of the isolates were resistant to third generation cephalosporins, cefotaxime, ceftazidime, and cefpodoxime. Sixty-five percent of the isolates were resistant to the monobactam drug aztreonam, 40.82% to ertapenem, and 31.36% to meropenem. Resistance to at least five antibiotics was observed in 38.46% of the isolates. Polymerase Chain Reaction (PCR) analysis of ESBL-encoding genes detected bla(CTX), bla(SHV), and bla(TEM) genes in 76.92%, 63.3%, and 44.37% of the isolates, respectively. Multiple ESBL genes were detected in majority of the isolates. The recently discovered New Delhi metallo-β-lactamase gene (bla(NDM-1)) was detected in two ESBL⁺ isolates. Our study shows that secondary contamination of fresh seafood with enteric bacteria resistant to multiple antibiotics may implicate seafood as a potential carrier of antibiotic resistant bacteria and emphasizes an urgent need to prevent environmental contamination and dissemination of such bacteria.201728867789
1122190.9999Antibiotic resistance profiles of gram-negative bacteria in southern Tunisia: Focus on ESBL, carbapenem and colistin resistance. The main objective of this cross-sectional study was to investigate the prevalence of beta-lactam (cephalosporins or carbapenems) or colistin resistant bacteria. Those were isolated from urine samples in two private polyclinics located in the Sfax region, in southern Tunisia. From September 2021 to August 2022, 116 strains resistant to β-lactams or colistin were isolated, identified by MALDI-TOF, and their antibiotic susceptibility was assessed by disk diffusion method. Resistance genes were detected by real-time PCR, standard PCR, and sequencing. The results revealed that the 116 strains consisted predominantly of Enterobacteriaceae (92.2 %) and non-fermenting bacteria (7.8 %). Among these strains, 21 (18.1 %) were resistant to carbapenems, three (2.7 %) to colistin, including two strains of Klebsiella pneumoniae (1.7 %) exhibiting resistance to both carbapenems and colistin. In Enterobacteriaceae, bla(CTX-A), bla(SHV), and bla(TEM) were found in 79.5 %, 46.7 %, and 40.2 % of strains, respectively. For these strains, the minimum inhibitory concentrations (MICs) of imipenem and ertapenem ranged from >32 to 6 μg/mL and > 32 to 2 μg/mL, respectively, with bla(OXA-48) and bla(NDM) detected in 21.7 % and 19.6 % of isolates, respectively. Seven A. baumannii isolates resistant to imipenem and meropenem (MICs >32 μg/mL and 8 μg/mL, respectively) carried bla(OXA-23) (n = 5) and bla(OXA-24) (n = 2). In addition, mutations in the mgrB gene conferring colistin resistance were identified in two isolates. Two K. pneumoniae were colistin-resistant and carried the bla(OXA-48) gene. These results highlight the urgency of developing new strategies for the identification and surveillance of pathogenic strains in humans to effectively combat this growing public health threat in Tunisia.202540553790