Multiplex PCR System for Rapid Identification of Bacterial Pathogens from Goats Presumed with Fever and/or Diarrhea

Abstract

Goat is one of the major livestock species, which play an important role in the economy of Bangladesh. However, outbreak of different infectious diseases in goats causes high mortality and economic losses due to lack of proper diagnosis and treatment. Conventional culture based methods for detecting pathogens are time consuming and laborious in compare to multiplex Polymerase Chain Reaction (mPCR), by which multiple pathogens can be detected at a time. The present study was aimed to perform faster molecular identification of multiple bacterial pathogens from whole blood of clinically infected goats. A total of 200 blood samples were collected from the goats that have fever and/or diarrhea at S. A. Quaderi Teaching Veterinary Hospital (SAQTVH) in Chittagong Veterinary and Animal Sciences University (CVASU) for the period of July-April 2017-2018. DNA was extracted and subsequently mPCR assay was performed for the screening of several bacterial pathogens (Salmonella spp., Listeria monocytogenes, Bacillus cereus, Yersinia enterocolitica, Campylobacter jejuni, Campylobacter coli, Clostridium perfringens, Vibrio cholerae, and Staphylococcus aureus). An antimicrobial susceptibility test (AST) against 10 antimicrobials for positive samples of each organism was conducted using the Kirby-Bauer Disc Diffusion Method on selective media. Staphylococcus aureus, Clostridium perfringens, Listeria monocytogenes and Salmonella spp. were detected from collected samples and their overall prevalence were 11.5%, 3.5%, 1% and 20.5%, respectively. Most common clinical signs were mild fever, nasal discharge, dyspnea and coughing (39.1%) for Staphylococcus aureus, diarrhea, convulsion, abdominal pain and incordination (57.1%) for Clostridium perfringens, fever, protrusion of tongue and incordination (100%) for Listeria monocytogenes and fever, anorexia, dehydration with mucus in stool (36.6%) for Salmonella spp infection in goats . Antimicrobial diagram represents Staphylococcus aureus showed resistance against Cefotaxime (74%), Cefixime (65%) and Tetracycline (65%) whereas highly sensitive against Amoxicillin (48%), Ciprofloxacilin (44%) and Gentamycin (44%). On the other hand Ampicillin (71%) and Gentamycin (71%) were highly resistant and Penicillin (57%) and Cefotaxime (57%) were highly sensitive against Clostridium perfringens infection. Penicillin (100%) and Cefixime (100%) were found sensitive against Listeria monocytogenes infection in goats. Salmonella spp. showed resistance to Ampicillin (78%) and Amoxicillin (59%) but sensitive to Ciprofloxacin (54%). This study may explore frequent clinical signs and symptoms of some common infectious diseases in goats and also determine the potential risk factors, measuring strength of association. In conclusion, rapid diagnostic methods like multiplex PCR were effective for diagnosis of multiple pathogens.