Listeria isolation ann molecular characterization from animal, food and environmental samples

In this study, listeria prevalence in sheep and cattle feeding Elazığ and it?s around investigated in February 2007- January 2008. Raw milk, cheese, water, animal food, feces, bulk milk swabs and environmental samples was collected from both farmhouses. Moreover, seasonal variation in listeria species was researched in those animals. To determinate genetic relationships between L. monocytogenes isolates was planned by Restriction Fragment Length Polymorphism (RFLP) and Random Amplified Polymorphic DNA (RAPD).Six cattle and 5 sheep farmhouses was used. Totally 719 samples as 415 bovine samples and as 304 sheep samples were cultured in this study. Total 106 milk (75 bovine, 35 sheep), 57 cheese samples (28 bovine, 29 sheep), 28 samples bulk milk swabs were performed. Furthermore, 72 fecal, 72 animal food, 72 water and 72 environment bovine samples and 60 fecal, 60 animal food, 60 water and 60 environment sheep samples were examined.Association of Official Agricultural Chemists/International Dairy Federation (AOAC/IDF) method in milk and cheese samples and United States Department of Agriculture/Food Safety and Inspection Service (USDA/FSIS) method in environmental and other samples were followed in this study. Isolated L. monocytogenes and other listeria strains were confirmed by Polymerase Chain Reaction (PCR) and biochemical reactions. Then, Restriction Fragment Length Polymorphism (RFLP) and Random Amplified Polymorhic DNA (RAPD) methods were performed amongst L. monocytogenes isolates. Furthermore, seasonal variations in listeria isolates were found out with some differency.Prevalence of Listeria spp. was found 6,4 % in whole samples. Eight of 46 isolates (17,4 %) L. monocytogenes, 18 of 46 (39,0 %) L. innocua, 5 of 46 (% 10,8) L. welshimeri, 8 of 46 (17,4 %) L. seeligeri and 7 of 46 (15,2 %) L. grayi were identified. L. ivanovii was not isolated in this study. listeria prevalence were found in cattle 9,4% and in sheep 2,4%.RAPD analyses were performed by two different primers (HLWL74 and HLWL85) and obtained 10 different profiles with 1-4 band patterns. In RFLP analysis by AluI and Tsp509I restriction endonucleases was obtained 4 different profiles and 2-3 bands patterns. According to obtained results in this study, RAPD analysis was found more discriminative than RFLP.Consequently, listeria existence in Elazığ and it?s around, the people of that region and animals were found to be as a risk factor. Moreover, higher proportion of cattle farms than sheep farms have been identified at risk. Results obtained that listeria that more transmitting to milk and milk products from animals than food during the preparation of environmental exposure is caused by listeria contaminations. Furthermore, seasonal variations indicate that autumn and spring more important than other seasons.