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    Detection of HEV RNA genotypes in amounts and Raw Milks Obtained from Different Animals
    (Ankara Microbiology Soc, 2019) Demirci, Mehmet; Yigin, Akin; Unlu, Ozge; Kilic Altun, Serap
    Hepatitis E virus (HEV) is one of the major foodborne viral pathogens transmitted through the fecal-oral route. Four genotypes of HEV are known to infect humans and it is reported that different types of HEV are active in zoonotic transitions. It is known that the HEV genotype 1 and HEV genotype 2 infections are generally acute and the HEV genotype 3 infections are chronic. Therefore, in the studies related to HEV infections, it is important to determine the genotypes to monitortreatment regimens. Although raw milk is often used in communities due to its low cost, there are limited data on the rates and the genotypes of HEV in our country and in the world. In light of this information, we aimed to investigate epidemiologically the quantity and genotypes of HEV RNA in 231 raw milk (48 cow milk, 65 goat milk, 65 sheep milk, and 53 donkey milk) samples. Viral RNAs were isolated from raw milk samples and the ORF2 region of HEV was investigated by the qRt-PCR method to determine quantitatively the presence of HEV RNA. In addition, among HEV RNA positive samples, the ORF2 region of HEV was amplified by nested PCR and the amplicons were sequenced. HEV RNA was detected in 47 (20.34%) raw milk samples, Positivity was detected in 14 (29.16%) of cow milk, 12 (18.46%) of goat milk, 8 of sheep milk (12.3) and 13 of donkey milk (24.5%). The amount of HEV RNA in cow milk found as the highest in both proportion and quantity. When the distribution of the HEV genotypes in the 47 positive samples was examined, 27 (57.44%) HEV genotype 1a, 10 (21.27%) HEV genotype 1 b, 4 (8.5%) HEV genotype 4c, 2 (4.2%) HEV genotype 3a, (2.13) HEV genotype lc, 1 (2.13%) HEV genotype 3e, 1 (2.13%) HEV genotype 3f and 1 (2.13%) HEV genotype 3g were determined. Although genotype la is more frequent, it has been revealed that different genotypes encountered in our country. In conclusion, it has been determined that HEV, one of the major foodborne viral agents, may be encountered in raw milk, and the genotypes that can cause infections in human are found especially in raw milk from animal sources. For the prevention of foodborne outbreaks, the presence of HEV in raw milk should not be ignored.
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    Detection of staphylococcal enterotoxin genes in raw milk samples by real-time PCR
    (Agricultural Research Communication Centre, 2019) Yigin, Akin; Demirci, Mehmet; Altun, Serap Kilic; Dinc, Hikmet
    Presence of significant level of enterotoxigenic S. aureus in raw milk of sheep, goat and donkey may cause serious food borne disease. Many people worldwide, use raw milk in their daily life but data on presence of virulence genes in cow, sheep, goat and especially donkey milk seems to be very limited. For this reason, aim of this study was to determine the presence both S. aureus and nine staphylococcal enterotoxin genes in cow, sheep, goat and donkey milks. A total of 231 raw milk samples were collected from 48 cow, 65 goat, 65 sheep and 53 donkey were collected. To detect presence of S. aureus both conventional culture and real-time PCR were used and to detect nine staphylococcal enterotoxin genes directly in milk, real-time PCR was performed. Conventional culture and real-time PCR results were found to be similar for presence of S. aureus and it was detected in 52 (22.51%) out of 231 raw milk samples. Staphylococcal enterotoxin genes were detected in 27 out of 52 S.aureus positive samples and a total of 62 enterotoxin genes were detected in these samples. However enterotoxin genes could not be detected in two S. aureus positive donkey milk. Hence, real-time PCR proved to be reliable and faster than conventional methods to detect presence of enterotoxigenic S. aureus in milk. Raw milk samples from different animals many contain enterotoxigenic S. aureus. Therefore, one should be careful during raw milk consumption as enterotoxigenic S. aureus in milk may cause dangerous public health problem which need routine screening for this pathogen in milk.
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    Presence of Staphylococcus aureus, staphylococcal enterotoxins and antimicrobial resistance in traditionally produced raw milk cheeses
    (M H Schaper Gmbh Co Kg, 2018) Gurbuz, Semra; Keskin, Oktay; Gurbilek, Sevil Erdenlig; Tel, Osman Yasar; Yigin, Akin; Demirci, Mehmet; Demir, Cemil
    The objectives of this study was to investigate the presence of Staphylococcus aureus, distribution of classical staphylococcal enterotoxin (SE) SEA to SEE, relevant gene/s and antimicrobial resistance pattern of S. aureus isolated from traditionally produced raw milk cheeses. A total of 106 fresh white cheese samples were examined. The 25 (23.6 %) of 106 cheese samples were found to be contaminated with coagulase positive staphylococci (CPS). From 52 isolates identified as S. aureus, one or more SEs was detected in 38.4 % of the isolates by ELISA whereas one or more se genes were detected in 50 % of the isolates by RT PCR. SEE (75 %) and see gene (61.5 %) were detected most frequently, whereas SED and sed gene were not detected in any isolates. Overall, 63.5 % of isolates were resistant to antimicrobial agents with 59.6 %, 13.5 %, 5.8 %, 5.8 % and 3.8 % of the isolates were resistant to penicillin, erythromycin, tetracycline, cefoxitin and kanamycin, respectively. The results of this study have revealed that cheeses made from raw milk were highly contaminated with S. aureus, therefore, creates a risk for public health due to the presence of enterotoxins as well as resistant strains against antimicrobial agents.