Detection of Biofilm Formation and Antibiotic Resistance in Klebsiella Oxytoca and Klebsiella Pneumoniae from Animal Origin Foods

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Biofilms are surface-attached microbial communities with distinct properties, which have a tremendous impact on our health and food safety. The study was aimed to detect biofilm formation and antibiotic resistance by Klebsiella oxytoca and Klebsiella pneumoniae from animal origin foods. In this study100 food samples were examined for the presence of Klebsiella oxytoca, Klebsiella pneumoniae, and other Enterobacteriaceae family members. In this study, 19 Klebsiella oxytoca and 5 Klebsiella pneumoniae isolates were isolated from cheese and minced meat samples using standard biochemical tests and identification kit. Biofilm formation in these isolates was detected by using microplate, Congo red agar, and tube adherence methods. Antibiotic susceptibility testing was performed using the disk diffusion method on Muller-Hinton agar. Using a microplate method strong biofilm formation was observed in 16 (84%) and 5 (100%) isolates of Klebsiella oxytoca and Klebsiella pneumoniae after 24 hours of incubation on Tryptic Soy Broth medium containing 2% of glucose respectively. After 24 and 48 hours of incubation on Tryptic Soy Broth without glucose strong biofilm formation was detected in 10 (52.6%) and 2 (40%) isolates of Klebsiella oxytoca and Klebsiella pneumoniae respectively. After 24 hours of incubation on Congo red agar 14 (73.7%) and 3 (60%), isolates of Klebsiella oxytoca and Klebsiella pneumoniae were slime factor positive respectively. In tube adherence method 13 (68.4%) and 4 (80%), isolates of Klebsiella oxytoca and Klebsiella pneumoniae were seen to have adhered strongly after 24 hours of incubation on Tryptic Soy Broth medium containing 2% of glucose. In general, strong biofilm formation by these strains was seen on Tryptic Soy Broth medium when supplemented with glucose. Among all the Klebsiella oxytoca, the highest rates of susceptibility were seen toward Trimethoprim-Sulfamethoxazole (100%) and Imipenem (94.7%) followed by Chloramphenicol (73.7%) and gentamicin (68.4%). Among 19 Klebsiella oxytoca isolates, the highest rates of resistance were seen in streptomycin (73.7%) and Kanamycin (73.7%) followed by ampicillin (63.2%). The majority of Klebsiella pneumoniae isolates were resistant to Kanamycin (80%) and Streptomycin (80%) followed by Amikacin (60%). On the other hand, 80% of Klebsiella pneumoniae were susceptible to imipenem, chloramphenicol, Trimethoprim-Sulfamethoxazole, and cefotaxime. Generally, majority of Klebsiella oxytoca, and Klebsiella pneumoniae isolates showed strong biofilm production on different growth conditions and majority of the isolates were also resistance for antibiotics. Therefore, biofilm production by these nosocomial bacteria has an implication public health and pave the way for increased resistance of biofilm-associated organisms to antimicrobial agents.