Statement of the Problem: Neonatal calf diarrhea (NCD) disease is an established zoonosis all over the world1. U.S. National Animal Health Monitoring System reported 57% of weaning calf mortality attributed to diarrhea. A limited research is reported about the fate and prevalence of multi-drug resistant bacteria and their resistance gene pool reservoir in NCD disease animals of Pakistan. The present study aimed to understand bacterial patho-biome and the antibiotic resistance potential in fecal ecology of NCD diseased animals.
Methodology, Theoretical Orientation & Findings: A total of 51 (morphologically different) bacterial strains were isolated from feces of bovine diarrheic animals up to 3 months of age, collected from various livestock farms located in different villages of district Okara, Faisalabad, Multan, Abbottabad, Muzafargarh and Islamabad Capital Territory (ICT) of Pakistan. On the basis of 16S rRNA gene sequence and phylogenetic analysis, the isolates belonged to 17 genera with a diverse prevalent rate (Accession numbers: LC488898 to LC488948 – Fig 1). Majority isolated strains (90 %) exhibited multi-drug resistance (against at least three antibiotics) with highest level of minimum inhibitory concentrations for Shigella spp. (MIC up to 70 µg/L) (Fig 2). Using PCR for Extended-spectrum β-lactamase (ESBL) and metallo-β-lactamase (MBL) producing bacteria, 60% isolates were positive for blaTEM antibiotic resistance gene whereas 55% were positive for blaNDM gene with the highest prevalence rate in Shigella spp.
Conclusion & Significance: These preliminary findings conclude the highest load of multi-drug resistant bacteria and antibiotic resistance genes, harbouring in diarrheic animal feces and pose the potential public health risk. These antibiotic resistance genes can transfer to other bacteria (horizontal gene transfer). Niche of neonatal calf diarrhea (NCD) animal feces is a major threat of zoonotic diseases and thus may be included in surveillance program of cattle infectious diseases in Pakistan. Results of these studies also showed the presence of candidate novel species of bacteria that may be validated as a new species from Pakistani ecology by poly-phasic taxonomic characterization.
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