Isolation and Molecular Detection of Listeria monocytogenes in Minced Meat, Frozen Chicken and Cheese in Duhok Province, Kurdistan Region of Iraq Saman Said Taha Said Ahmed * , Bizhar Ahmed Tayeb, Alind Mahmood Ameen, Sowaila Mekaeel Merza and Yousif Hamed Mohammed Sharif * Laboratory Department, Veterinary Directorate in Duhok 1, 1000 Jn Galy Way, 42001 Duhok Province, Iraq * Corresponding author: Saman Said Taha Said Ahmed, Laboratory Department, Veterinary Directorate in Duhok 1, 1000 Jn Galy Way, 42001 Duhok Province, Iraq, E- mail: shokri.mustafa@uqconnect.edu.au Received date: February 20, 2017; Accepted date: March 07, 2017; Published date: March 13, 2017 Copyright: © 2017 Ahmed SSTS, et al. This is an open-access article distributed under the terms of the creative commons attribution license, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Abstract Aims: Listeria monocytogenes (L. monocytogenes) is the food-borne pathogen responsible for listeriosis. It is considered a serious public health risk, and is spread through the consumption of food products. The disease can be fatal to humans and animals. The objective of the present study was to isolate and identify L. monocytogenes from frozen chicken, minced meat, and cheese in Duhok province, Iraq. Methodology and Results: Between March and October 2015, equal numbers (n=50) of samples of minced meat, frozen chicken, and cheese were collected (total n=150). Biochemical and microbiological tests, including real- time PCR, were performed to determine the prevalence of L. monocytogenes in the samples. Out of 150 samples, 20 displayed greyish/black colonies with black halos on Oxford and Palcam agar. of these putative Listeria, 12 were subsequently identified as L. monocytogenes by PCR. L. monocytogenes was detected in 1 (2%), 7 (14%), and 4 (8%) of isolates from cheese, minced meat, and frozen chicken, respectively. Conclusion: Significance and Impact of study: Detection of pathogenic bacteria in foods, such as those analysed in this study, is crucial for safeguarding public health. The qPCR technique offers a rapid and reliable method for isolation of many foodborne pathogens in different kinds of foods. Keywords: Listeria monocytogenes; Real time PCR; Identifcation; Detection; Food Introduction Listeria spp. are broadly distributed in the environment. Tey can be isolated from soil and water Jersik, et al. [1]. A wide range of animal species can become contaminated by L. monocytogenes, including mammals, domesticated animals, pets, fsh, birds, and crustaceans Alsheikh et al. [2]. In mammals, L. monocytogenes can cause premature births and meningoencephalitis Aznar et al. [3]. L. monocytogenes is a concern to public health because of its capability to survive or even grow under harsh conditions [4,5]. Te increasing rate of L. monocytogenes involvement in food-born outbreaks has led to the requirement for a rapid method for testing food products. Almost all cases of listeriosis tend to be food-born, and a number of food items can become contaminated by L. monocytogenes, including raw chicken meat, raw minced meat, sof cheese, and fsh. Most of these items are widely consumed in Iraq’s Kurdistan and Duhok provinces. A number of molecular biological methods have been described for detection of L. monocytogenes, including DNA probes and PCR techniques. Direct detection of L. monocytogenes in food products by PCR has been reported in several cases Bansal et al. [6], Manzano et al. [7], Agersborg et al. [8], O’Connor et al. [9] and Hudson et al. [10]. To our knowledge, there is no published data regarding L. monocytogenes prevalence in food samples from Duhok province. In fact, there are no ofcial data of food-born listeriosis recorded in Duhok, as L. monocytogenes is rarely tested for in food samples. Te objective of the present research was to assess the use of PCR for the detection of L. monocytogenes in food products, and to determine the prevalence of Listeria spp. in food products in Duhok, by using molecular and biochemical methods. Materials and Methods Samples (n=150) of diferent food products, including chicken (n=50), minced meat (n=50), and cheese (n=50), obtained from diferent supermarkets, restaurants and veterinary quarantine centres in Duhok province, were tested. All samples had been correctly stored, and were placed in separate sterile plastic bags to prevent spilling and cross contamination. Te samples were brought to the laboratory on crushed ice, and were kept at 4°C until testing (within 4 h). Food samples were analysed for the presence of Listeria spp. using a selective enrichment and isolation protocol, as recommended by the United States Department of Agriculture (USDA) [11]. Microbiological investigation Samples (25 g) were added to 225 mL of ½ strength Fraser broth (Conda, Spain), to obtain a 1:10 dilution. All samples were homogenized (30-60 sec) and incubated (30°C, 24 h). From this primary enrichment, 0.1 mL was then inoculated into 10 mL of Fraser broth, and incubated (37°C, 48 h) with shaking. A loopful of the subsequent culture was streaked on the surface of diferent agars (PALCAM listeria agar (Conda, Spain) with supplement, and OXFORD agar (Lab UK) with X122 supplement). Agar plates were then incubated (37°C, 48 h) and observed (at 24 to 48 h incubation) Ahmed et al., J Food Microbiol Saf Hyg 2017, 2:1 DOI: 10.4172/2476-2059.1000118 Research Article Open Access J Food Microbiol Saf Hyg, an open access journal ISSN:2476-2059 Volume 2 • Issue 1 • 1000118 Journal of Food: Microbiology, Safety & Hygiene J o u r n a l o f F o o d : M i c r o b i o l o g y , S a f e t y & H y g i e n e ISSN: 2476-2059