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Applying Modern Technique of qPCR Coupling with Propidium Monoazide to Detect Enterotoxigenic Staphylococcus aureus in Cream Pastry Products
2020
Azizkhani, Maryam | Tooryan, Fahimeh
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.
Показать больше [+] Меньше [-]Potential risk of some pathogens in table eggs
2020
EL-Kholy, A.M. | Saadia H. EL-Shinawy | Hemmat Seliem | Mohamed M.A. Zeinhom
This study was conducted to record the potential risk of some pathogens in table eggs in Beni-Suef city, Egypt. A total of 100 table eggs samples (Farm and Baladi eggs) were randomly collected from poultry farms, markets, supermarkets and groceries in Beni-Suef city, 50 farm eggs samples (each of 3 eggs) and 50 baladi eggs samples (each of 5 eggs) were examined for the presence of coliforms, E.coli, Salmonella, coagulase positive Staphylococcus aureus and Staphylococcal enterotoxins. Isolates were identified by biochemical, serological and molecular (PCR) methods. The obtained results in the present study revealed that (22%) of the examined samples were contaminated with Coliforms. The other identified genera were Citrobacter freundii, Citrobacter diversus, Edwardsiella tarda, Enterobacter spp., Morgonella morganii, Klebsiella oxytoca, Providencea spp., Serratia fonticola and Yersinia intermedia. E.coli (atypical type) was detected in a percentage of 27.27%. True fecal E. coli and Salmonella spp. failed to be detected in any of the whole examined 100 eggs samples. Additionally, Staphylococcus spp. was detected with incidence rate of (13%), out of them, 8 (61.5%) isolates were accounted for coagulase positive Staphylococcus aureus. Enterotoxin profiling revealed that two classical enterotoxin genes (SEA and SED) were detected either singly or in combination. The potential health hazards and the suggested control measures of the isolated strains had been discussed.
Показать больше [+] Меньше [-]Prevalence and characterisation of class 1 and 2 integrons in multi-drug resistant Staphylococcus aureus isolates from pig farms in Chongqing, China
2020
Ye, Chao | Hou, Fengqing | Xu, Dongyi | Huang, Qingyuan | Chen, Xia | Zeng, Zheng | Peng, Yuanyi | Fang, Rendong
Integrons are mobile DNA elements that allow for acquisition and dissemination of antibiotic-resistance genes among pig farm-derived bacteria. Limited information is available on integrons of Staphylococcus aureus from pig farms. The aim of this study was to characterise and investigate the prevalence of class 1 and 2 integrons in multi-drug resistant (MDR) S. aureus isolates from pig farms. A total of 724 swabs were collected from 12 pig farms in Chongqing, China, and examined by conventional microbial and molecular methods. In total, 68 isolates were S. aureus, 57 of which were methicillin resistant (MRSA). All 68 isolates were MDR strains and carried integrons, of which 88.2% (60/68) harboured both class 1 and 2. In addition, 85.3% (58/68) of the class 2 integron-positive isolates carried the β-lactam resistance gene (blaTEM₋₁), and 66.7% (40/60) of the class 1 integron–positive isolates carried the aadA1c, aadA1 or dfrA1 gene for respective streptomycin and spectinomycin or trimethoprim resistance. Class 1 and 2 integrons are common among the pig farm-derived S. aureus isolates. On account of their significance for public health, the prevalence of the integrons and their associated resistance genes in pig farm-derived S. aureus isolates should be paid special attention.
Показать больше [+] Меньше [-]Characterisation of classical enterotoxins, virulence activity, and antibiotic susceptibility of Staphylococcus aureus isolated from Thai fermented pork sausages, clinical samples, and healthy carriers in northeastern Thailand
2020
Sankomkai, Wanwisa | Boonyanugomol, Wongwarut | Kraisriwattana, Kairin | Nutchanon, Julalak | Boonsam, Kraisorn | Kaewbutra, Sasalux | Wongboot, Warawan
Contamination by Staphylococcus aureus of food produced from animal sources may have diverse and multifactorial causes that depend on geographical distribution. The goal of this study was to isolate and characterise S. aureus strains from contaminated fermented pork sausage, which is a local food of northeastern Thailand. S. aureus strains were isolated from local pork sausage, and the presence of classical enterotoxins was determined by PCR and reversed passive latex agglutination. These results were compared with strains derived from hospitalised patients and healthy carriers. Additionally, production of extracellular enzymes and haemolysin, biofilm formation, and antibiotic susceptibility were assessed. S. aureus was identified in 36 sausage isolates (60%). The strains positive for staphylococcal enterotoxin A were more frequently found in isolates from sausage and healthy carriers than in those from patients. All tested S. aureus strains were positive for DNase, lipase, proteinase, haemolysin, and biofilm formation; notably, strains isolated from food and healthy carriers displayed similar values. Most isolates were resistant to penicillin and ampicillin, while none were to methicillin. Thai fermented pork sausages are associated with a high risk of staphylococcal food poisoning, which may be linked to contamination caused by carriers. Dissemination of knowledge regarding best practices in sanitation and hygiene is important in local communities.
Показать больше [+] Меньше [-]Histologic changes and gene expression patterns in biopsy specimens from bacteria-inoculated and noninoculated excisional body and limb wounds in horses healing by second intention
2020
Jorgensen, Elin | Hjerpe, Freja B. | Hougen, Hans P. | Bjarnsholt, Thomas | Berg, Lise C. | Jacobsen, Stine
OBJECTIVE To evaluate histologic changes and gene expression patterns in body and limb wounds in horses in response to bacterial inoculation. SAMPLE Wound biopsy specimens from 6 horses collected on days 7, 14, 21, and 27 after excisional wounds (20 wounds/horse) were created over the metacarpal and metatarsal region and lateral thoracic region (body) and then inoculated or not inoculated on day 4 with Staphylococcus aureus and Pseudomonas aeruginosa. PROCEDURES Specimens were histologically scored for the amount of inflammation, edema, angiogenesis, fibrosis organization, and epithelialization. Quantitative PCR assays were performed to quantify gene expression of 10 inflammatory, proteolytic, fibrotic, and hypoxia-related markers involved in wound healing. RESULTS Except for gene expression of interleukin-6 on day 27 and tumor necrosis factor-α on day 14, bacterial inoculation had no significant effect on histologic scores and gene expression. Gene expression of interleukin-1β and -6, serum amyloid A, and matrix metalloproteinase-9 was higher in limb wounds versus body wounds by day 27. Gene expression of cellular communication network factor 1 was higher in limb wounds versus body wounds throughout the observation period. CONCLUSIONS AND CLINICAL RELEVANCE The lack of clear markers of wound infection in this study reflected well-known difficulties in detecting wound infections in horses. Changes consistent with protracted inflammation were evident in limb wounds, and gene expression patterns of limb wounds shared similarities with those of chronic wounds in humans. Cellular communication network factor warrants further investigation and may be useful in elucidating the mechanisms underlying poor limb wound healing in horses.
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