Staphylococcus aureus Genotypes of Subclinical Bovine Mastitis Milk in the Middle Western Anatolia (original) (raw)
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International Journal of One Health, 2021
Background and Aim: Milk is a chief source of many nutrients. However, we must also bear in mind that it is a potential source for many cases of food poisoning. This study was conducted to investigate the prevalence of cow mastitis and evaluate the presence of enterotoxins and antibiotic resistance patterns in Staphylococcus aureus isolated from milk and contact humans in El-Behira Province, Egypt. Materials and Methods: A total of 680 milk samples from 170 cows and 86 human samples consisting of 43 hand swabs and 43 nasal swabs were analyzed. The milk samples were subjected to the California mastitis test. Results: The general occurrence was 23.1% (157/680) where 48 quarters had clinical mastitis and 109 had subclinical mastitis. Subsequently, S. aureus was isolated in Baird-Parker agar where typical and atypical colonies were selected and submitted to coagulase and complementary tests. Out of 48 samples of mastitic milk studied, 16 (33.3%) showed contamination by S. aureus whereas 109 samples of subclinical mastitis showed contamination in only 18 (16.5%). On the opposite hand, of the 86 human samples, 33 revealed S. aureus contamination, corresponding to 38.37% of the samples. Furthermore, multiplex polymerase chain reaction targeting nuc and the staphylococcal enterotoxin-encoding genes sea, seb, sec, sed, and see were performed after culture, revealing that 88.2% (30/34) of milk samples and 93.9% (31/33) of human samples were variably positive to those genes. Conclusion: The use of nuc gene based PCR is an accurate and rapid method for S aureus isolates detection. A high prevalence of multiple drug-resistant isolates of S. aureus recovered from both human and milk represents further evidence for possible veterinary hazards as well as public hazards, especially to those that consume milk from this region.
The objective of the present study was to investigate the epidemiological and genetic relationships of classical enterotoxins of S. aureus in goat’s raw milk, meat and food handlers in Toukh city in Qaluobia governorate, Egypt. A total of 100 goat, s raw milk and meat samples (50 of each) were collected from randomly distributed herds in streets for buying milk and in public markets for peddler meat. Hand and nasal swabs were collected from milkers and butchers (30 of ech). All samples were subjected for bacteriological examination for isolation and identification of S. aureus. Isolates were underwent reversed passive latex agglutination technique for detection of enterotoxigenic S. aureus. A multiplex PCR assay could successfully amplify the diagnostic DNA bands of 270bp, 165bp, 69bp and 306bp of genes for staphylococcal enterotoxins A, B, C, and D respectively. PCR was applied on the serologically identified 16 (20.25%) isolates out of 79 S. aureus which isolated from the examined goat’s food samples and human handlers by using one universal forward and reverse primers, specific for each individual toxin gene. None of the samples was positive for SEE indicating the zoonotic and genetic relationships
Comparative Clinical Pathology, 2012
Staphylococcus aureus is a major causative pathogen of clinical and subclinical mastitis of dairy domestic ruminants. This organism produces a variety of extracellular toxins and virulence factors such as enterotoxin SEI and SEJ that contribute to its pathogenic potential. In this study 25 S. aureus isolates obtained from four dairy herds of Urmia region which is located in West Azerbaijan province in Iran. The tested isolates were identified on the basis of the cultural and biochemical properties, as well as amplification of the aroA gene which is specific for S. aureus. All isolates were also analyzed for the presence of the SEI (sei) and SEJ (sej) encoding genes using polymerase chain reaction (PCR). Seven positive isolates were detected for sei, but sej gene was not detected in any of the total number of 25 isolates. The present study revealed that the PCR amplification of the aroA gene could be used as a powerful tool for identification of S. aureus from the cases of bovine mastitis. Results of the present study also showed that the strains of S. aureus which cause mastitis can potentially produce enterotoxin SEI. Overall, our results suggest that it is of special importance to follow the presence of enterotoxin-producing S. aureus in other dairy products, especially for protecting the consumers from staphylococcal food poisoning.
International Journal of Food Microbiology, 2011
Staphylococcus aureus is a major foodborne pathogen and it has the ability to produce a number of extracellular toxins. We analyzed 1070 food samples obtained from retail markets and dairy farms in the Marmara Region of Turkey for the presence of S. aureus. Out of 147 isolates, 92 (62.6%) were enterotoxigenic. PCR was used to investigate the presence of staphylococcal enterotoxin genes (sea, seb, sec, sed, see, seg, seh, sei, sej, sek, sel, sem, sen, seo, sep, seq and seu), exfoliative toxin genes (eta and etb) and the toxic − shock syndrome toxin gene (tst). The PCR results showed that 53.3% of the isolates contained staphylococcal enterotoxin-like (SEl) toxin genes (seg, seh, sei, sej, sek, sel, sem, sen, seo, sep, seq and seu) which were more frequent than classical enterotoxin genes (sea to see). Furthermore, seo, sei, sem, seg, seu and sec were found in 37.0, 32.7, 30.4, 29.3, 29.3 and 27.2% of the isolates, respectively. The tst gene was detected and confirmed by DNA sequencing in 9 isolates. The presence of eta and etb were not found in the isolates. Enterotoxigenic capabilities of isolates with SEA-SEE were investigated by ELISA. Enterotoxigenic S. aureus isolates produced one to three enterotoxins, with the most frequently produced types being enterotoxin A and C. There was a correlation of 72.1% between production of a specific toxin and the presence of the respective genes. PFGE analysis was used to identify genetic-relatedness of enterotoxigenic S. aureus isolates and the results revealed that 13 groups of isolates from different or the same origin that contained the same genes showed 100% homology with indistinguishable band patterns. The other enterotoxigenic isolates showed related band patterns with 72-86% homology in sea-, 61-90% homology in sec-, 80-96% homology in seh-, and 69-96% homology in sep-positive isolates. To our knowledge, this is the first study to examine enterotoxins and related gene contents of S. aureus food isolates in the Marmara Region of Turkey.
Prevalence and Genetic Characterization of S. Aureus Strains Isolated from Raw Milk and Its Products
Suez Canal Veterinary Medicine Journal. SCVMJ
Raw milk and milk products could be contaminated with bacterial pathogens such as S.aureus, which could be transmitted to human causing severe illness. In order to throw the light on the prevalence as well as molecular typing of S.aureus isolated from raw milk and milk, a total of 202 samples of raw milk (n=122) and milk products (n=80) were collected from Ismailia, Port-said and Suez Governorates, Egypt. The collected samples were subjected to bacteriological examination. The prevalence of S. aureus was (14%) in raw milk samples and (5%) in milk products samples. Antimicrobial susceptibility for isolated strains was carried out where (85.7%) of the isolated strains were sensitive to Azithromycin. PCR protocol was used for detection of Spa, Coa, Sea, Seb, Sec, Sed and See genes. Spa gene and Coa gene were detected in (90%) and (80%) of the tested S.aureus strains, respectively. Concerning S.aureus enterotoxins genes, 40% of the tested strains were positive for Sed gene, while 30% were positive to See gene. The combination of both phenotypic and genotypic analysis is a valuable diagnostic tool for identification of S.aureus in raw milk and its products.
Journal of Veterinary Science, 2004
In the present study, 35 Staphylococcal strain isolated from milk samples of 16 cows from eight farms of three different geographic locations in Central Java, Indonesia, and from milk samples of 19 cows from 19 farms of different geographic locations in Hesse, Germany, were compared pheno-and genotypically. On the basis of cultural and biochemical properties as well as by amplification of the 23S rRNA specific to Staphylococcus aureus, all isolates could be identified as S. aureus. In addition, all S. aureus isolates harboured the genes clfA and coa encoding staphylococcal clumping factor and coagulase, and the gene segments encoding the immunoglobulin G binding region and the X-region of protein A gene spa. By PCR amplification, the genes seb, seg, seh, and sei was observed for the S. aureus cultures isolated in Central Java, Indonesia and the genes sec, sed, seg, seh, sei, sej and tst for the S. aureus cultures isolated in Hesse, Germany. None of the S. aureus of both origins harboured the genes sea, see, eta and etb. All isolates were additionally positive for the genes nuc, fnbA, hla, and set1. The gene hlb was found for 6 cultures from Central Java, Indonesia and 16 cultures from Hesse, Germany. However, the gene fnbB and the gene segments cnaA and cnaB were not present among the strains isolated in Central Java, Indonesia and rare among the strains isolated in Hesse, Germany. It was of interest that most of the S. aureus isolated in Central Java, Indonesia harboured the gene cap5 and most of the strains isolated in Hesse, Germany the gene cap8. The phenotypic and genotypic results of the present study might help to understand the distribution of prevalent S. aureus clones among bovine mastitis isolates of both countries and might help to control S. aureus infections in dairy herds.
Journal of Human Environment and Health Promotion, 2019
"Background: Staphylococcus aureus is one of the major causes of food poisoning. Since milk is a nutritious source of proteins and vitamins, it could provide the optimal conditions for the growth of several bacterial pathogens, such as S. aureus. The present study aimed to assess the frequency of Staphylococcus aureus classical enterotoxin genes in raw milk samples in Zanjan, Iran. Methods: In total, 82 bovine, unpasteurized milk samples were collected from the dairy farms in various rural areas in Zanjan, Iran. The isolation and identification of S. aureus were performed using the Baird-Parker agar, routine biochemical tests, and polymerase chain reaction (PCR) targeting the S. aureus-specific femA gene. In addition, staphylococcal enterotoxin genes (e.g., sea, seb, sec, sed, and see) were assessed using PCR. Results: Following the appearance of yellow colonies with yellow zones on Mannitol salt agar, 21 S. aureus isolates (25.6%) were detected. In total, 80.9% of the isolates were positive for the presence of SE genes, and the most frequent SE gene was sea (88.2%), followed by see (58.8%), and seb (52.9%). Furthermore, 76.5% of the isolates had two or more SE genes simultaneously. Conclusion: According to the results, the presence of enterotoxigenic S. aureus in the studied raw milk samples confirmed the possible risk posed on the public health. Therefore, it is recommended that the quality of dairy product quality programs be optimized in order to intensify the sanitary inspection of these products."
Aim and Background: Staphylococcus aureus is a major human pathogen that also causes important infections in cattle and sheep. The present study aimed to test genetic diversity among strains of S. aureus isolated from cattle (n=34) and humans (n=22) by DNA typing. Materials and Methods: Fluorescent amplified fragment length polymorphism (FAFLP) is the genotyping tool used in the study. The presence of the mecA and Panton-Valentine leukocidin (PVL) genes among these strain groups was also checked. Results: A dendrogram deduced from FAFLP showed that all the strains clustered into 10 groups (A-J) with a relative genetic divergence of less than 8%. Sixty-seven percent of the isolates from bovine sources clustered together in two clades (A and H), while another major cluster with 13 isolates (59%) (Cluster G) had all strains from a human host. The remaining strains from both the hosts clustered independently into smaller clusters with the exception of two strains of human origin, which clustered along with a bovine cluster. Thirteen strains belonging to cluster G were highly clonal. About 77% of strains obtained from human infections were methicillin-resistant S. aureus (MRSA), whereas only 29% of strains from bovine origin were MRSA. Only three strains from human origin showed PVL positive, while no strain from cattle had PVL genes. The complete absence of PVL genes in all the bovine strains in the study appears to be significant. Conclusions: FAFLP can be successfully applied to assess the genetic relationship of S. aureus isolates from different hosts. The study also provided the valuable epidemiological data on S. aureus from bovine sources in India, which is lacking.
Acta Microbiologica et Immunologica Hungarica, 2011
Staphylococcus aureus is considered one of the most important food borne pathogens.A total of 111 isolates of S. aureus were cultured from raw milk samples during January 2009 to June 2009 from Tehran and Mashhad. The coagulase gene polymorphism and the prevalence of classical enterotoxin genes of S. aureus strains were determined by PCR-RFLP (restriction fragment length polymorphism) and Multiplex-PCR. Disk diffusion method was used to determine the susceptibility of isolates to antimicrobial agents as instructed by Clinical and Laboratory Standards Institute.Sixty-seven % of the isolates harboured one or more enterotoxin genes. The most prevalent gene was sec, found in 59 % of the isolates. Approximately 8% of the isolates were positive for sea, seb and sed genes. Only one isolate had see gene. The rate of coexistence of enterotoxin genes was 14%. All S. aureus isolates were susceptible to ciprofloxacin, gentamicin, imipenem, minocycline, oxacillin and vancomycin. They were resist...