First substantiation of clinico-molecular investigation of pathogenic Listeria monocytogenes in Nili-Ravi buffaloes

Background: Listeriosis is a zoonotic disease of humans, animals, birds, fish, and crustaceans worldwide. Domestic animals, especially ruminants, are more susceptible to listeriosis. This infectious disease is caused by Listeria monocytogenes, an intracellular bacterium that can cross blood-brain, placental and intestinal barriers. In Pakistan, the incidence and reliable diagnostic tools for the L. monocytogenes are unidentified in Nili-Ravi buffaloes. Aims: This study was designed to inspect listeriosis in buffaloes through molecular techniques and haemato-biochemical analyses. Methods: A total of 230 samples (115 milk and 115 faecal samples) were collected from symptomatic listeriosis cases in Nili-Ravi buffaloes of 3 geographical districts (Rawalpindi, Faisalabad, and Muzaffargarh) Punjab, Pakistan. These samples were processed for DNA extraction using commercialized kits, and L. monocytogenes was confirmed by conventional PCR. Results: The results revealed that 6.08% and 4.34% of the isolates from milk and faecal samples were found positive for L. monocytogenes, respectively. The phylogenetic analysis of these isolates showed 97100% similarity to isolates from the USA, Switzerland, Japan, and India. The accession numbers on NCBI GenBank appeared as HF558398 (Switzerland), KP965732 (India), EU372032 (USA), and LC259850 (Japan). Haemato-biochemical examinations showed that the values of WBCs, plasma fibrinogen, ALT, and AST significantly increased (P<0.05) in diseased buffaloes compared to healthy ones. Conclusion: The occurrence of listeriosis in buffaloes urges continuous monitoring and surveillance to prevent this emerging disease in Pakistan.