Isolation, identification, and characterization of Listeria spp. from various animal origin foods.

Deepti N Nayak,C V Savalia,I H Kalyani,Rajeev Kumar,D P Kshirsagar

doi:10.14202/vetworld.2015.695-701

Abstract

Aim:The present study was undertaken with the prime objective of isolating and identifying Listeria spp. from various foods of animal origin sold at retail market outlets in the city of Navsari, Gujarat.Materials and Methods:Total 200 samples comprising of milk, milk products, meat, and fish (50 each) collected aseptically from local market which were subjected first to pre-enrichment in half strength Fraser broth followed by enrichment in full strength Fraser broth and subsequent plating on PALCAM agar. The growth with the typical colony characteristics were further identified up to species level on the basis of their morphological and biochemical characteristics. Cultures identified as Listeria monocytogenes were further subjected to in vitro pathogenicity tests and detection of different virulence-associated genes viz. actA, hlyA, and iap using polymerase chain reaction.Results:Of the total 200 food samples of animal origin; 18 (9%) were found positive for Listeria spp. which were identified as Listeria seeligeri (6, 33.3%), Listeria innocua (5, 27.7%), Listeria welshimeri (4, 22.2%), and L. monocytogenes (3, 16.6%). The highest prevalence was observed in milk samples (8). Species wise, 6 isolates of L. seeligeri which included two each from cow milk, buffalo milk, and meat samples; 5 L. innocua isolates included four recovered from fish and one from meat sample; 4 L. welshimeri comprised of two isolates from ice cream and one each from buffalo milk and meat sample; and 3 isolates of L. monocytogenes recovered from milk (1 cow and 2 buffalo milk). All 3 L. monocytogenes isolates screened for the presence of virulence genes viz. actA, hlyA, and iap using the specific primers revealed the presence of all the genes suggesting the possibility of danger of foodborne listeriosis among raw milk consumers.Conclusion:Listeria spp. was isolated from 9% (18/200) of the animal origin food samples viz.; milk, milk products, meat, and fish with the highest prevalence in the milk samples. L. monocytogenes was isolated from 3 milk samples only. L. seeligeri was the predominant species isolated followed by L. innocua, L. welshimeri, and L. monocytogenes in this study. L. monocytogenes were found to carry virulence genes like actA, hly A, and iap genes suggesting the pathogenic potential of these isolates.

Highlights

The organisms of genus Listeria are Grampositive, facultative anaerobic, non-spore-forming, rod-shaped bacteria with a low G + C content
6 isolates of L. seeligeri which included two each from cow milk, buffalo milk, and meat samples; 5 L. innocua isolates included four recovered from fish and one from meat sample; 4 L. welshimeri comprised of two isolates from ice cream and one each from buffalo milk and meat sample; and 3 isolates of L. monocytogenes recovered from milk (1 cow and 2 buffalo milk)
All 3 L. monocytogenes isolates screened for the presence of virulence genes viz. actA, hlyA, and iap using the specific primers revealed the presence of all the genes suggesting the possibility of danger of foodborne listeriosis among raw milk consumers

Summary

Introduction

The organisms of genus Listeria are Grampositive, facultative anaerobic, non-spore-forming, rod-shaped bacteria with a low G + C content. L. monocytogenes is a primary human pathogen amidst rare reports of illnesses caused by L. seeligeri, L. ivanovii, and L. innocua [4]. The fatality ranges from 30% to 75% especially in high risk groups like pregnant women, unborn or newly delivered infants, and the elderly people as well as persons with severe underlying disease conditions like immune-suppression, AIDS, chronic conditions like cirrhosis [5,6]. This organism is halotolerant, and can grow and multiply at refrigeration temperatures [7]

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