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The global emergence of extended-spectrum beta-lactamase (ESBL)-producing Klebsiella pneumoniae presents a significant public health threat and complicates antibiotic treatment for infections. This study aimed to determine the prevalence of ESBL-producing K. pneumoniae in a clinical setting, analyze their antimicrobial susceptibility profiles, and characterize both phenotypic and genetic determinants. A total of 507 non-duplicate clinical isolates of Enterobacterales were collected between 2019 and 2020, and third-generation cephalosporin resistance was screened by disk diffusion. Identification of K. pneumoniae was confirmed using biochemical tests and PCR with species-specific primers. Antimicrobial susceptibility testing was conducted using disk diffusion, and phenotypic ESBL production was confirmed using the combined disk method. Multiplex PCR detected ESBL genes (blaTEM, blaSHV, and blaCTX-M) and identified blaCTX-M groups. The genetic relatedness of ESBL-producing strains was assessed using the ERIC-PCR approach. Fitty-four isolates were confirmed as ESBL producers, all classified as multidrug-resistant (MDR). All ESBL-producing K. pneumoniae isolates exhibited resistance to ampicillin and cefotaxime, with high resistance rates for ciprofloxacin (98.2%), azithromycin (94.4%), piperacillin-tazobactam (88.9%), and trimethoprim (83.3%). Genotypic analysis revealed blaCTX-M was present in 94.4% of isolates, blaSHV in 87%, and blaTEM in 55.5%. The blaCTX-M-1 group was the most prevalent, accounting for 96.1% of isolates. Co-harboring of blaCTX-M, blaSHV, and blaTEM occurred in 42.6% of isolates, with co-carrying of blaCTX-M, and blaSHV was observed in 23/54 isolates. The ERIC-PCR analysis revealed 15 distinct types, indicating high genetic diversity. These findings highlight the urgent need for ongoing monitoring to control the spread of ESBL among K. pneumoniae and emphasize the importance of early detection and appropriate antibiotic selection for effectively treating infection caused by these pathogens.
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Background and Aim: Bovine mastitis is an inflammation of the mammary gland of dairy cattle that causes economic losses due to poor quantity and quality of milk. The extensive or incorrect use of antibiotics has increased in the veterinary field, leading to the emergence of antibiotic-resistant pathogens worldwide. This study aimed to investigate bovine mastitis bacterial pathogens in Sakon Nakhon, Thailand. Materials and Methods: A total of 35 dairy farms were screened for clinical and subclinical mastitis using the California Mastitis Test and clinical examination. Polymerase chain reaction was used to characterize bacterial species-induced mastitis (380 isolates) in cattle and antimicrobial resistance genes, and disk diffusion and broth microdilution were used to characterize antimicrobial susceptibility. Results: The prevalence of Staphylococcus epidermidis (38.10%; 32/84)-induced mastitis in cattle was considerably high, followed by Streptococcus agalactiae (33.33%), Streptococcus uberis (25%), Klebsiella pneumoniae (8.33%), and Staphylococcus aureus (4.76%). In this study, Staphylococcus spp. isolates demonstrated 100% susceptibility to cefoxitin, and no antibiotic-resistance genes were identified. Tetracycline (TET) and macrolide-resistant genes of Streptococcus spp. revealed that tetM was predominant in 55.63% (79/142), followed by tetS + erm(B) (16.90%). Antibiotic susceptibility tests revealed the following resistance profiles to bacterial species: TET (85.92%), clindamycin (29.58%), erythromycin (15.49%), levofloxacin (14.08%), and penicillin (0%). Gram-negative bacterial isolates (K. pneumoniae [8.33%], Klebsiella variicola [2.38%], Klebsiella quasipneumoniae [1.19%], and Escherichia coli [1.19%]) were recovered and still susceptible to meropenem (100%), ceftazidime (97.06%), ceftriaxone (79.41%), and ciprofloxacin (79.41%). Conclusion: This result suggested that mastitis pathogens in this area were susceptible to most antimicrobials, with the exception of streptococci against TET. In this study, limited data were available including one from small-holder dairy farms and study only dairy farms in Sakon Nakhon, Thailand. So, more farms should be included in the future studies.
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Background and Aim: Over recent years, antimicrobial-resistant Klebsiella species in humans, animals, food animals, food products, and agricultural environments have been the center of attention due to its role in the evolution of antimicrobial resistance. The emergence of resistance to fluoroquinolones and cephalosporins of third and higher generations in Klebsiella oxytoca has not received much attention in animal husbandry compared to that in Klebsiella pneumoniae. Reports on K. oxytoca are limited in the study area. Therefore, we investigated the antimicrobial susceptibility and resistance genes in K. oxytoca isolated from slaughtered pigs in Thailand. Materials and Methods: Microbiological examination was conducted on 384 Klebsiella spp. isolates recovered from slaughtered pigs in ten provinces of Thailand. Seventy-two K. oxytoca isolates (18.75%) were examined for antimicrobial-resistant genes (ß-lactamase [bla TEM, bla CTX-M, and bla SHV]) and fluoroquinolone-resistant genes (qnrA, qnrB, qnrC, qnrD, qnrS, oqxAB, aac(6')-Ib-cr, and qepA). Results: The most common genotype was bla CTX-M (58/72, 80.55%), followed by bla TEM with bla CTX-M (7/72, 9.72%) and bla TEM (6/72, 8.33%). The most common bla CTX-M group was bla CTX-M-1 (19/58, 32.76%), followed by bla CTX-M-9 (1/58, 1.72%). Plasmid-mediated quinolone resistance genes were identified in 13 (18.05%) isolates: qnrS (16.70%) and qnrB (1.4%). All 13 isolates had qnrS transferable to an Escherichia coli recipient, whereas qnrB was not detected in any transconjugants. Either bla CTX-M or bla TEM harbored by one K. oxytoca strain was transferable to an E. coli recipient. Analysis of antimicrobial susceptibility revealed that more than 90% of the bla CTX-M-carrying K. oxytoca isolates were susceptible to chloramphenicol, trimethoprim, ceftazidime, cefepime, cefotaxime, amoxicillin-clavulanic acid, piperacillin-tazobactam, and fosfomycin. All K. oxytoca isolates (13) harboring qnr were susceptible to carbapenem and ceftriaxone; however, 43 (74.13%) of the K. oxytoca isolates harboring bla CTX-M exhibited extended-spectrum ß-lactamase activity. Most of the K. oxytoca isolates from pigs were highly resistant to ampicillin, azithromycin, and gentamicin. Conclusion: To prevent further transmission of Klebsiella spp. Between food animals and humans, strict control of antibiotic use in clinical and livestock settings is necessary along with routine disinfection of the livestock environment and efforts to increase awareness of antimicrobial resistance transmission.
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Dissemination of the mobile colistin resistance gene mcr in Enterobacterales among humans, animals, and the environment is a public health issue. We characterized mcr genes in the Klebsiella pneumoniae complex (KpnC) isolated from slaughtered pigs in Thailand. The 280 KpnCs consisted of K. pneumoniae (85%), Klebsiella quasipneumoniae (8.21%), and Klebsiella variicola (6.79%). mcr genes were detected in 6.79% (19/280) of KpnC isolates, consisting of mcr-8 (n = 9; 3.21%), mcr-7 (n = 7; 2.50%), mcr-7 + mcr-8 (n = 2; 0.71%), and mcr-1 + mcr-7 (n = 1; 0.36%). K. pneumoniae predominantly carried the mcr-7 and mcr-8 genes, while K. variicola and K. quasipneumoniae harbored mcr-7 and mcr-8, respectively. Six of the nineteen mcr-harboring KpnC isolates exhibited colistin resistance, and five had mcr-1 or mcr-8 transferable to an Escherichia coli recipient. Antimicrobial susceptibility analysis revealed that all mcr-carrying KpnC isolates were susceptible to carbapenems, cefotaxime, cefepime, amoxicillin/clavulanic acid, piperacillin/tazobactam, amikacin, and fosfomycin, and had high resistance to azithromycin. Multilocus sequence analysis demonstrated that the mcr-harboring KpnC isolates were genetically diverse. A 'One-Health' approach is useful to combat antimicrobial-resistant bacteria through coordinating the human, animal, and environmental sectors. Hence, continuous monitoring and surveillance of mcr-carrying KpnCs throughout the pork supply chain is crucial for ensuring public health.