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The emergence of carbapenem-resistant organisms (CROs) has become a great challenge alarming the public health community. Thus, the current study was conducted to investigate the occurrence of CROs among pet animals suffering from respiratory illness. Nasal swabs from 100 pet animals (51 dogs and 49 cats) showing respiratory illness were screened for CROs. The obtained swabs were streaked onto CHROMagar mSuper CARBATM medium followed by sub-culturing on MacConkey agar. Colonies were identified by cultural characteristics, Gram staining, and biochemical tests as well as molecular techniques. Then, the identified isolates were confirmed to be CROs after being non-susceptible to at least one of four carbapenem antibiotics using the Kirby-Bauer method. The confirmed CRO isolates were also investigated on molecular basis for carbapenemase-encoding genes (blaNDM, blaKPC, blaOXA-48, blaVIM, blaIMP). Out of 100 pet animals, 6 yielded CROs with an overall occurrence rate 6% (3.9% for all tested dogs and 8.2% for all tested cats). The obtained CRO isolates were Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Enterobacter cloacae with the following distribution among the examined animals 3%, 1%, 1%, and 1%, respectively. BlaNDM was the only detected carbapenemase-encoding gene among the isolates and it was detected in two cat isolates. In conclusion, the results of the current study highlight the emergence of carbapenem-resistant organisms among pet animals suffering from respiratory illness which may have a possible public health risk. [ABSTRACT FROM AUTHOR]

Background & Objectives: Klebsiella pneumoniae is a significant cause of nosocomial infections. This bacterium survives in difficult conditions by forming biofilms in hospital equipment and causes severe infections. On the other hand, the emergence and spread of carbapenem resistance among bacteria and biofilm production is a current health concern. There are controversial findings about the relevance of this issue. This study aimed to evaluate the relationship between biofilm formation and carbapenem resistance among clinical isolates. Materials & Methods: A total of 160 isolates of Klebsiella pneumoniae were collected. Molecular methods were used to detect resistance genes. Subsequently, the ability to produce biofilms in isolates with resistance genes was assessed. Finally, the correlation of biofilm formation among resistant isolates was calculated using χ2 test. Results: 79 imipenem-resistant isolates were obtained. 46 isolates (66.66%) containing VIM gene, 36 isolates (52.17%) containing OXA-48 gene, five isolates (7.24%) containing NDM gene, Six isolates (8.69%) containing gene IMP and five isolates (7.24%) also had KPC gene. The results showed a significant correlation between the ability to form biofilms and the presence of carbapenem-resistant genes. Conclusion: Increased carbapenem resistance in Klebsiella pneumoniae isolates and its association with biofilm formation is severe warning for basic measures to combat this phenomenon.

Carbapenemase-producing Klebsiella pneumoniae (CP-Kp) is a real global health threat. Environmental reservoirs of resistance gene determinats, such as effluents of hospital wastewaters, are acquiring increased relevance in the selection of plasmid-encoded carbapenemase genes. The presence of Hg in environmental reservoirs may exert a positive selective pressure on tolerant bacteria, favoring the co-transfer of carbapenemase genes and mer operons. In our study, 63 CP-Kp isolates were screened for mer operons by whole genome sequencing (MySeq). Conjugation assays were performed with 24 out of 63 CP-Kp isolates harboring the mer operon. Ten transconjugants (Tc-Kp) were selected with Hg. Plasmid DNA of Tc-Kp was extracted and sequenced using single-molecule real-time (SMRT) technology (PacBio, Sequel II system) with later annotation. Plasmid analysis revealed that Tc-Kp from blaIMP-like (n = 3) showed a single plasmid belonging to IncC group with two complete mer operon next to blaIMP-like. Tc-Kp from blaVIM-1 (n = 2) harbored two plasmids, one with blaVIM-1 in an IncL, and mer operon was in an IncFIB plasmid. Tc-Kp from blaOXA-48-like (n = 5) showed 2 plasmids. blaOXA-48-like was found in an IncL plasmid, whereas mer operon was (i) in an IncR plasmid associated with blaCTX-M-15 in 3 Tc-Kp-OXA-48-like, (ii) in an IncC plasmid associated with blaCMY-2 in 1 Tc-Kp-OXA-48-like, (iii) and in an IncFIB plasmid associated with blaCTX-M-15 in 1 Tc-Kp-OXA-48-like. This is, to our knowledge, the first study to describe in K. pneumoniae producing plasmid-encoded carbapenemase, the potential impact of Hg in the co-transfer of mer operons and carbapenemase genes located in the same or different plasmids. Key points: • Environmental reservoirs are playing an important role in the selection of carbapenemase genes. • Conjugation assays, selecting with Hg, obtained 10 transconjugants with carbapenemase genes. • mer operons were located in the same or different plasmids than carbapenemase genes. [ABSTRACT FROM AUTHOR]

Background & Objectives: Klebsiella pneumoniae is a significant cause of nosocomial infections. This bacterium survives in difficult conditions by forming biofilms in hospital equipment and causes severe infections. On the other hand, the emergence and spread of carbapenem resistance among bacteria and biofilm production is a current health concern. There are controversial findings about the relevance of this issue. This study aimed to evaluate the relationship between biofilm formation and carbapenem resistance among clinical isolates. Materials & Methods: A total of 160 isolates of Klebsiella pneumoniae were collected. Molecular methods were used to detect resistance genes. Subsequently, the ability to produce biofilms in isolates with resistance genes was assessed. Finally, the correlation of biofilm formation among resistant isolates was calculated using χ2 test. Results: 79 imipenem-resistant isolates were obtained. 46 isolates (66.66%) containing VIM gene, 36 isolates (52.17%) containing OXA-48 gene, five isolates (7.24%) containing NDM gene, Six isolates (8.69%) containing gene IMP and five isolates (7.24%) also had KPC gene. The results showed a significant correlation between the ability to form biofilms and the presence of carbapenem-resistant genes. Conclusion: Increased carbapenem resistance in Klebsiella pneumoniae isolates and its association with biofilm formation is severe warning for basic measures to combat this phenomenon. [ABSTRACT FROM AUTHOR]