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Klebsiella pneumoniae is an important pathogen that causes pneumonia and bloodstream infections, especially in neonates and intensive care patients. The carbapenems remain an important therapeutic option for clinicians, particularly against cephalosporin-resistant Gram-negative pathogens. This increased use of carbapenems at clinics has resulted in the evolution and spread of carbapenem-resistant K. pneumoniae . In this study, we isolated six bla K. pneumoniae carbapenemase (KPC)-producing strains belonging to sequence type 258 (ST258) from clinical, environmental, and veterinary sources. Antibiotic susceptibility was performed on these isolates and the genes responsible for extended-spectrum beta-lactamases and metallo-beta-lactamase production were screened. The molecular typing was done using multilocus sequence typing. Isolated strains were resistant to various antibiotic classes, including carbapenems, and carried the carbapenem-resistant gene, blaKPC. All strains were susceptible to tigecycline and colistin. This is the first report detecting K. pneumoniae ST258 strains in Pakistan.

Presence of multiple drug resistant pathogens in hospital waste is a serious public health concern, because it may ultimately be disseminated to the human. Current study was designed with the objective to estimate the occurrence of ESBL-producing K. pneumoniae in hospital settings and waste. For this purpose, cross sectional study for a period of one year was designed and non-probability sampling techniques was used to collect total n=112 samples from various sample sources of hospital waste including ward waste, operation theatre waste, wastewater and hospital sludge. Isolation of the K. pneumoniae was done by using selective agar, biochemical identification of the isolates was done through API 20E kit (bioMérieux, France). Molecular identification of the isolates was done by amplifying 16SrDNA with PCR. According to CLSI guidelines disc diffusion assay was performed for antibiotic susceptibility profiling. PCR of MDR isolates was done for the molecular detection of various ESBL genes. Results of the study showed 17 (15%) percentage prevalence of MDR K. pneumoniae from all 112 collected samples. Among various sample sources wastewater showed the highest percentage (23%) prevalence of MDR K. pneumoniae. In 17 confirmed isolates bla CTX-M and blaCTX-M1 were found in 13 (76%) and 12 (71%) respectively which showed the highest prevalence as compared to all other investigated genes. While bla TEM , bla SHV , bla NDM-1 and bla OXA-48 were found with percentage prevalence 9 (53 %), 1 (6%), 9 (53%) and 6 (35 %) respectively. Whereas bla KPC , bla IMP , bla VIM and bla GIM were not detected in any of the isolate. Taking together, strict rules and regulations should be adopted at public as well as hospital level to restrict the dissemination of antibiotic resistance from hospital environment to humans.

Tamoor Hamid Chaudhry,1,2 Bilal Aslam,1,2 Muhammad Imran Arshad,3 Roman Farooq Alvi,2 Saima Muzammil,2 Nafeesa Yasmeen,4 Muhammad Aamir Aslam,3 Mohsin Khurshid,2 Muhammad Hidayat Rasool,2 Zulqarnain Baloch1 1Biomedical Research Center, Northwest Minzu University, Lanzhou 730030, People’s Republic of China; 2Department of Microbiology, Government College University Faisalabad, Faisalabad, Pakistan; 3Institute of Microbiology, University of Agriculture Faisalabad, Faisalabad, Pakistan; 4College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, People’s Republic of ChinaCorrespondence: Bilal Aslam; Zulqarnain Baloch Email drbilalaslam@gcuf.edu.pk; znbalooch@yahoo.comIntroduction: Intense livestock farming practices enforcing the farmers to use antibiotics as food supplements on a routine basis. Aberrant use of antibiotics is associated with the emergence of antibiotics resistance and resistant superbugs. Keeping in view the current scenario, the present study was designed for the first time from Pakistan with a specific aim to estimate the prevalence of the carbapenem-resistant Klebsiella pneumoniae in veterinary settings and the waste in Pakistan.Methods: A total of 138 samples from various veterinary sources were collected by employing a nonprobability sampling technique. Isolation and phenotypic identification of carbapenem-resistant K. pneumoniae were performed according to the CLSI standard. Molecular detection of various antibiotic resistance genes (ARGs) was done through PCR by using specific primers against each ARG. According to the pasture scheme, the multilocus sequence typing (MLST) was performed to characterize the K. pneumoniae sequence types (STs).Results: According to the results of the study, overall 9.4% (13/138) isolates were confirmed carbapenem-resistant K. pneumoniae. Among various carbapenem ARGs particularly, the blaNDM-1 was found in 92.3% (12/13) isolates followed by blaOXA-48 84.6% (11/13). MLST results revealed that overall 3 STs were found in the study which includes ST29, ST11, and ST258. Taking together, this is the first study to our best knowledge which demonstrated the prevalence of carbapenem-resistant K. pneumoniae and its various STs prevalent in veterinary settings and the waste of Pakistan.Conclusion: Based on the above-mentioned facts, we suggested that veterinary settings and waste are the potential source and reservoir of carbapenem-resistant K. pneumoniae, which may be disseminated to the environment and ultimately can affect the public and companion livestock health.Keywords: veterinary settings, carbapenem resistance, Klebsiella pneumoniae, Pakistan