scholarly journals Determination of the prevalence of Salmonella spp. and S. aureus in meat products by Real-Time PCR and testing their antibiotic susceptibility

2021 ◽  
Vol 77 (06) ◽  
pp. 6533-2021
Author(s):  
REYHAN IRKIN ◽  
BERKAY BOZKURT ◽  
GULENDAM TUMEN
Keyword(s):  
Public Health ◽  
Staphylococcus Aureus ◽  
Risk Factor ◽  
Real Time ◽  
Real Time Pcr ◽  
Meat Products ◽  
Penicillin G ◽  
Diffusion Method ◽  
Salmonella Spp ◽  
And Staphylococcus Aureus

From a public health point of view meat products contain high pathogenic risk factors. This is because they can be contaminated with Salmonella spp. and Staphylococcus aureus microorganisms, and, more importantly, antibiotic resistance has been reported in these microorganisms at an increasing frequency. To examine the presence of Salmonella spp. and Staphylococcus aureus in samples of raw and semi-cooked (chicken doner, meat doner, chicken, beef, and lamb products) meat from markets of Izmir and Balikesir, Turkey were analysed. The presence of microorganisms in the samples was determined by Real-Time PCR method using Salmonella spp. and S. aureus specific primers. Following Real-Time PCR, microorganisms were isolated by selective culture methods and biochemical tests from the positive meat samples and tested for their antibiotic susceptibility using the Kirby-Bauer Disk Diffusion Method. The antibiotic disc diffusion method showed that S. aureus was resistant to penicillin G, oxytetracycline, sulfamethoxazole, tetracycline, erythromycin, and ampicillin, whereas Salmonella spp. was resistant to penicillin G, sulfamethoxazole, erythromycin, and ampicillin. As these products are consumed frequently, their contamination with S. aureus (≥ 5 × 103 cfu/g) and Salmonella spp. can be a risk factor for food poisoning. The contamination of meat products’ with S. aureus and Salmonella spp. can be a risk factor for public health and the antibiotics to be preferred in illness treatment are of critical importance.

A Rapid Multiplex Real-Time PCR High-Resolution Melt Curve Assay for the Simultaneous Detection of Bacillus cereus, Listeria monocytogenes, and Staphylococcus aureus in Food

2016 ◽  
Vol 79 (5) ◽  
pp. 810-815 ◽  
Author(s):  
FEREIDOUN FORGHANI ◽  
SHUAI WEI ◽  
DEOG-HWAN OH
Keyword(s):  
Staphylococcus Aureus ◽  
Listeria Monocytogenes ◽  
High Resolution ◽  
Bacillus Cereus ◽  
Real Time ◽  
Real Time Pcr ◽  
Simultaneous Detection ◽  
Cost Savings ◽  
High Resolution Melt ◽  
And Staphylococcus Aureus

ABSTRACTThree important foodborne pathogens, Bacillus cereus, Listeria monocytogenes, and Staphylococcus aureus, are of great concern for food safety. They may also coexist in food matrices and, in the case of B. cereus and S. aureus, the resulting illnesses can resemble each other owing to similar symptoms. Therefore, their simultaneous detection may have advantages in terms of cost savings and rapidity. Given this context, a rapid multiplex real-time PCR high-resolution melt curve assay for the simultaneous detection of these three pathogens in food was developed. The assay successfully detected B. cereus (gyrB), L. monocytogenes (hly), and S. aureus (nuc) in a single reaction, and the average melting temperatures were 76.23, 80.19, and 74.01°C, respectively. The application of SYTO9 dye and a slow melt curve analysis ramp rate (0.1°C/s) enabled the production of sharp, high-resolution melt curve peaks that were easily distinguishable from each other. The detection limit in food (milk, rice, and lettuce) was 3.7 × 103 CFU/g without an enrichment step and 3.7 × 101 CFU/g following the 10-h enrichment. Hence, the assay developed here is specific and sensitive, providing an efficient tool for implementation in food for the simultaneous detection of B. cereus, L. monocytogenes, and S. aureus.

scholarly journals Bacterial pathogens isolated from currency notes circulating amongst live-bird marketers in Damaturu and Potiskum, Yobe State, Nigeria

2021 ◽  
pp. 25-32
Keyword(s):  
Public Health ◽  
Staphylococcus Aureus ◽  
Antibiotic Susceptibility ◽  
Bacterial Pathogens ◽  
Diffusion Method ◽  
Salmonella Spp ◽  
Bacillus Spp ◽  
Klebsiella Spp ◽  
Live Bird ◽  
Coagulase Positive Staphylococcus

Bacterial contamination of currency notes is of veterinary and public health importance because contaminated notes could serve as vector for the spread of pathogenic and perhaps multidrug resistant bacteria. The aim of this study was to assess the antibiotic susceptibility profile of bacterial contaminants found in various denominations of the Nigerian currency circulating among live-bird vendors in Yobe State, Nigeria. Three hundred and twenty (320) currency notes of all denominations were collected from the marketers for investigation. All samples were screened for bacterial pathogens according to standard techniques. The disc diffusion method was used to assess the antibiotic susceptibility of each of the isolated bacterial species against twelve antimicrobial drugs. The result showed that the higher denominations (N1000 - N100) were contaminated by Bacillus spp, (48.2 %), Eischerchiia coli ((13.5 %), Klebsiella spp (6.4 %) Pseudomonas aeruginosa (5.0%), Salmonella spp (0.7%), Coagulase positive Staphylococcus aureus (17.0%), and Streptococcus spp (9.2%) while the lower denominations (N50 - N5) were contaminated by Bacillus spp (37.1%), coagulase positive Staphylococcus aureus (19.1%,), E. coli (14.6 %), Klebsiella spp (5.1 %), P. aeruginosa (6.7%), Salmonella spp (7.3%) and Streptococcus spp (10.1%). All the isolated bacteria were resistant to ampicillin, oxacillin, amoxicillin, and tetracycline. Ciprofloxacin had the greatest activity followed by nitrofurantoin, neomycin, gentamicin, chloramphenicol and streptomycin. The present study revealed that Naira notes circulating among live-bird marketers were contaminated by pathogenic bacteria. It is recommended that live-bird traders should observe strict personal and environmental hygiene while engaging in their daily transaction to forestall any public health threat that may arise from transmission of disease pathogens from the legal tender of transaction in the market.

scholarly journals Epidemiological survey on the prevalence of Salmonella spp. in the Sardinian pig production chain, using real-time PCR screening method

2019 ◽  
Vol 8 (2) ◽  
Author(s):  
Carlo Pala ◽  
Tiziana Tedde ◽  
Sara Salza ◽  
Maria Teresa Uda ◽  
Stefano Lollai ◽  
...  
Keyword(s):  
Salmonella Typhimurium ◽  
Multiplex Pcr ◽  
Real Time ◽  
Real Time Pcr ◽  
Meat Products ◽  
Processed Meat ◽  
Finishing Pigs ◽  
Salmonella Spp ◽  
Production Chain ◽  
Pig Production

The aim of this study was to evaluate the prevalence of Salmonella spp. in the Sardinian pig production chain in order to establish the incidence of monophasic serovariant of Salmonella Typhimurium on isolates with molecular methods (real-time PCR and multiplex PCR). Samples were collected in three EC slaughterhouses, four small slaughterhouses annexed to farmhouses, one meat distribution center, four meat cutting laboratories and four sausage processing plants. A total of 166 samples were collected and analyzed: 46 environmental samples, 48 finishing pigs, 16 piglets, 24 samples of non-processed meat, 28 meat preparations and 4 meat products. All samples were processed with an initial screening using the real-time PCR MicroSEQ® Salmonella spp detection Kit (Applied biosystems, life technologies) and with the TaqMan® Real-time PCR to confirm the kit results. Samples that tested positive for Salmonella spp were confirmed with cultural method using the standard ISO 6579. Positive samples were submitted to phenotypic identification. One colony from each positive sample was serotyped with multiplex PCR method. Salmonella spp was isolated in 7 on 166 samples (4.22 %). Among the positive samples, two came from finishing pigs, two belonged to the category meat preparations, two to meat products, one was an environmental sample. Multiplex PCR confirmed that the collected strains belonged to the species Salmonella Typhimurium (1), Salmonella derby (3) and monophasic serovariant of Salmonella Typhimurium (3).

scholarly journals Development of SYBR Green Real time PCR for identification methicillin-resistance Staphylococcus aureus (MRSA)

2013 ◽  
Vol 7 (1) ◽  
pp. 67-71
Author(s):  
Noor Hashim Kareem ◽  
Majeed Arsheed Sabbah ◽  
Anas Noori Ibraheem ◽  
Al-Shayma’a Muhammad Said
Keyword(s):  
Staphylococcus Aureus ◽  
Real Time ◽  
Real Time Pcr ◽  
Methicillin Resistance ◽  
Sybr Green ◽  
Diffusion Method ◽  
Clinical Samples ◽  
Duplex Pcr ◽  
Minimum Concentration ◽  
Specific Identification

The increasing resistance of staphylococci to ß -lactam antibiotics has become a major clinical problem. Development of rapid and sensitive techniques for detection of MRSA is an important aim for public health. A duplex PCR were established for specific identification of methicillin-resistance Staphylococcus aureus (MRSA) in clinical samples. In this work a duplex SYBR Green real time PCR was developed for rapid identification of MRSA in local methicillin-resistance S. aureus isolates. Twenty methicillin-resistance S. aureus isolates, as determined by disc diffusion method, were subjected to DNA extraction and PCR amplification. Two genes were amplified successfully, mecA (533bp) and femA (314bp), as targets for methicillin-resistance and specific identification of S. aureus, respectively using conventional PCR. Sensitivity of the duplex PCR showed that the minimum concentration of DNA that gave positive results for the two genes was 30ng/µl. In order to develop rapid and sensitive test for identification of MRSA, serial dilutions of purified DNA were amplified gradually according to their concentrations using SYBR Green real time PCR. These results indicated that the SYBR Green real time PCR can be used for identification of methicillin-resistance S. aureus (MRSA) in clinical

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