BACKGROUND: Staphylococcus aureus is one of the most important human pathogens that cause infection and also food intoxication by secreting various enterotoxins. Conventional culturing methods to detect S. aureus have some limitations such as being time-consuming due to bacterial growth and low precision and sensitivity in detecting viable but non-cultivable cells. OBJECTIVES: The objective of this study was to detect and quantify enterotoxigenic (A-E) S. aureus in cream pastry products applying PCR coupling with propidium monoazide (PMA) to distinguish dead and live cells. METHODS: One hundred samples were randomly collected from pastry shops in Amol, in a period of 2 months. After preparing dilutions, bacterial pellets were separated and treated with PMA before DNA extraction. Real time PCR was conducted in order to quantify S.aureus cells and enterotoxigenic strains using specific primers. RESULTS: Results of conventional method were close to PMA-qPCR data (P>0.05), but data from qPCR that includes live and dead cells shows more bacterial count than two other methods. Sensitivity of the method applied in the present study, detecting low number of S.aureus cells (less than 10/g) seems considerable. CONCLUSIONS: Findings showed that applying PCR coupling with PMA results in more reliable data than conventional culturing method. Regarding this approach being time-effective, considerably sensitive and specific, it is expected that it be used in food quality control labs in monitoring systems in future.

BACKGROUND: Staphylococcus aureus is one of the most important pathogenic microorganisms in meat products, especially those that are repeatedly handled by hand in the production process. Beta-lactam drugs, especially new generations of Cephalosporins, are used for treatment of most infections that are caused by Staphylococcus aureus. But the production of beta-lactamase enzyme by some strains has led to the failure for treating the infections that are associated with this organism. OBJECTIVES: The present study was conducted to evaluate the prevalence and comparison of the antimicrobial effect of methanolic extracts of red pepper and red onion on Staphylococcus aureus with beta-lactamase gene that was isolated from minced meat in Semnan city. METHODS: For this reason, sampling from 30 distribution and supply centers of packaged meat in Semnan city was performed in hygienic conditions and all of the samples were tested for presence of Staphylococcus aureus with beta-lactamase gene by biochemical methods and molecular confirmation by PCR assay. Also, the antibacterial effect of red pepper and red onion extracts on these isolates was evaluated by minimum inhibitory concentration (MIC), minimum bacterial concentration (MBC), well distribution and bacterial growth curve tests. RESULTS: The results showed that 16.6 percent of samples were contaminated with Staphylococcus aureus with beta-lactamase gene. Red pepper and red onion extracts had good antibacterial effects on these isolates and in all the tests, the red pepper extract was more effective than the red onion extract. CONCLUSIONS: By proving stronger antimicrobial effect of red pepper, it is recommended to use pepper in sufficient amounts along with onion in foods that are made from minced meat like all kinds of Kebab.

BACKGROUND: Subclinical mastitis plays an important role in the economic losses of dairy cattle farms. Staphylococcus aureus is one of the most important causes of this disease. Treatment of this disease with synthetic antibiotics has complications like antibiotic resistance. Using herbal antibiotics can be an excellent way to reduce these side effects.OBJECTIVES: This study aimed to determine the minimum inhibitory concentration of alcoholic extract of olive leaf on Staphylococcus aureus isolated from milk of cows with subclinical mastitis to achieve herbal treatment.METHODS: This study was conducted on 175 Holstein female cattle. The milk samples of 60 cows were obtained with the sterilized method, and Subclinical mastitis-positive cases were determined using the California mastitis test. Staphylococcus aureus bacteria were isolated from positive samples by culture method, and the minimum inhibitory concentration of olive (Olea europaea L.) leaves alcoholic extract on isolated bacteria was determined by microdilution method.RESULTS: From 175 cows under study, 60 cows had a positive California mastitis test, and Staphylococcus aureus separated from milk samples of 14 cows. The minimum inhibitory concentration of olive (Olea europaea L.) leaves extract on this bacterium was 12000 ppm.CONCLUSIONS: Alcoholic extract of olive (Olea europaea L.) leaves has an antimicrobial effect on Staphylococcus aureus as a cause of mastitis. The minimum concentration required for this effect was 12000 ppm. Further studies on the impact of this plant on other bacterial causes of subclinical mammary inflammation in cows and investigation of the effective substances in the extract are needed.

BACKGROUND: Staphylococcus aureus is one of the most important pathogenic bacteria for human that is easily transferred during slaughtering, processing, packaging, storage and handling of meat and meat products as a result of poor hygienic principles, and causes staphylococcal food poisoning. Objectives: The objective of this study was to evaluate the contamination of raw and cooked ground beef in retail shops of Mazandaran to S. aureus and also detection of enterotoxin-producing genes in the isolates. Methods: One-hundred fifty ground beef samples (95 raw and 65 cooked) were collected randomly from retail shops, 21 May-21 July 2017. S. aureus was counted via culturing on Baird Parker Agar medium. Detection of enterotoxins A-E and G, H, I and J producing genes was conducted applying real-time PCR technique. Results: 68% of samples showed S. aureus contamination. The average count in raw and cooked ground beef samples was 3.1×105 cfu/g and 5.7×103 cfu/g, respectively. From 92 S. aureus isolates, 23 isolates (25%) were carrying enterotoxin coding genes; amongst them 15 isolates (65.2%) were carrying just a single gene and the rest more than one gene. Two isolates carrying SEA+ SEC, two isolates SEA+SEE, one isolate SEA+SEG, one isolate SEC+SEI, one isolate SEA+SEC+SEG and one isolate SEE+SEG. Conclusions: These results show that enterotoxigenic S. aureus strains are present on considerable numbers in retail ground meat in Mazandaran.

BACKGROUND: Staphylococcus aureus (S. aureus) possess a variety of virulence genes that are involved in the pathogenesis of infections caused by this agent. OBJECTIVES: The aim of this study was to evaluate the prevalence of the genes encoding collagen binding (cna) and fibronectin binding (fnb) adhesion factors in Staphylococcus aureus isolates from food and clinical specimens. METHODS: During the time period 2010-2013, a total of 38 isolates of Staphylococcus aureus from clinical specimens and 32 isolates from food samples were collected. All isolates were identified biochemically and subjected to DNA extraction. The accuracy of DNA extraction from each isolate was confirmed by PCR amplification of aroA gene and then the presence of cna and fnb genes in the extracted DNAs was assessed by PCR, using the specific primers. RESULTS: The results showed that among 38 isolates from clinical specimens, 15 (39.5%) & 32 (84.2%) and among 32 isolates of food origin 10 (31/2%) and 16 (50%) isolates had cna and fnb genes respectively. Thirteen clinical (34.21%) and 6 food isolates (18.7%) were positive for both fnb and cna genes and 4 clinical (10.5%) and 12 food isolates (37.5%) were lacking both genes. CONCLUSIONS: According to the results, it is concluded that, although these adhesion genes are not present in all Staphylococcus aureus clinical isolates, but their prevalence is high and using the products of these genes in vaccines may be effective in protecting against infections caused by this bacterium.

An intramammary device (IMD) was adapted for use in ewes; this device was made of abraded poly. ethylene material (1.7 mm in diameter, 47 mm long) and formed a 15-mm-diameter loop in the gland cistern. The IMD was inserted in 1 gland in each of 43 ewes. A significant (P < 0.0001) increase in milk somatic cell count (SCC) was observed in glands provided with an IMD. This increase was attributable to an increase in neutrophil numbers and was observed during the first 12 weeks after insertion. The IMD had a protective effect against experimentally induced staphylococcal mastitis (Staphylococcus aureus and S epidermidis), although different milk SCC were required for protection from each bacterial species in most ewes (10(6) and 2 X 10(5) cells/ml, respectively). Histologic studies revealed that the IMD induced local squamous metaplasia in the glandular part of the lactiferous sinus. Erythrocytes were found in milk from glands provided with an IMD throughout the studied period (35 days of the 45-day lactation) and, in some cases, blood clots were observed during the first 2 weeks of lactation. Glands with IMD also had lower milk production and quality at 30 and 32 days of lactation. Eight ewes with IMD were studied throughout a subsequent lactation. Milk from the IMD-containing glands had an increase in SCC, as in the previous lactation period; did not contain blood clots or erythrocytes; and had normal composition (similar to that in glands without the IMD).

Six primiparous Holstein cows were fed a Se-deficient diet, beginning at least 90 days before their first calving, and 6 other primiparous cows were given the same diet plus a supplement of 2 mg of Se/cow/d as sodium selenite. All cows were fed their diets for the duration of the experimental period. One uninfected quarter of each cow was injected with 25 microgram of Escherichia coli endotoxin at postpartum week 5. Leukocytes were isolated by centrifugation from milk collected at postinjection hour 16. Isolated cells were 92 +/- 3% neutrophils and were incubated with Staphylococcus aureus or E coli in a 1:300 ratio. Phagocytosis and intracellular killing by neutrophils were assessed after 0, 30, 60, and 90 minutes by a fluorochrome assay, using acridine orange. Viability of neutrophils was assessed by use of trypan blue. Superoxide anion production and hydrogen peroxide production by neutrophils also were determined. Cows fed Se-deficient diets had significantly (P < 0.05) lower blood Se concentration and blood glutathione peroxidase activity than cows fed Se-supplemented diets. Selenium status had no effect on the phagocytic capacity of neutrophils. Neutrophils obtained from cows fed Se-supplemented diets killed a significantly (P < 0.05) higher percentage of ingested bacteria than did neutrophils from cows fed the Se-deficient diet. Viability was significantly (P < 0.05) reduced by incubation with S aureus in neutrophils from both groups of cows, with neutrophils from Se-deficient cows having lower viability. Superoxide anion production did not differ significantly between neutrophils from the 2 groups, but extracellular hydrogen peroxide concentration was significantly (P < 0.05) higher in neutrophils harvested from milk of cows fed the Se-deficient diet.

A study was conducted to examine sources of variation introduced into a phagocytosis assay as a result of the isolation of neutrophils from bovine blood, including variation attributable to isolation of neutrophils from blood, variation between duplicate determinations of percentage phagocytosis, and the variation in the ability of neutrophils isolated from blood (over repeated collections from the jugular vein) to phagocytose. For the phagocytosis assay, jugular venous blood from each of 4 cows was divided into 2 equal portions. The neutrophils were isolated by lysis of red blood cells with 0.2% sodium chloride. The neutrophils (2 X 10(7)) were incubated in duplicate with 32P-labeled Staphylococcus aureus ([32P]SA; 2 X 10(8)) inskimmed milk samples (2.5% final concentration) prepared from 4 cows. This process was repeated thrice on neutrophils isolated from 4 cows at 2-week intervals. The proportions of variation in percentage of 32P-labeled S aureus phagocytosed between duplicate neutrophil isolations and between duplicate assay determinations were 0 and 1%. Differences among skimmed milk sources and among runs, using blood neutrophils taken at different times from the same donor cow, accounted for 62 and 36% of the total variation. The results indicated that variation arising from blood neutrophil isolation introduced into a phagocytosis assay within a single-day trial is of no concern. The large variation among skimmed milk sample sources indicated differences among cows in the ability of their milk to support phagocytosis. The variation in neutrophil isolations over time for any cow was considered too large to allow for evaluation of physiologic and environmental effects on phagocytosis of neutrophils isolated from blood.