Isolation and Identification of Coagulase Positive Staphylococcus aureus in Cow Row Milk in Tehran by Culture and Real-Time PCR Methods
Subject Areas :
Jalal Shayegh
1
,
Sahba Mohamadzadeh
2
1 -
2 -
Keywords: Coagulase positive, Staphylococcus aureus, raw milk, Tehran, real-time PCR,
Abstract :
Coagulase-positive Staphylococcus aureus (S. aureus) is one of the main causes of contamination of raw milk and dairy products, which can lead to food poisoning in humans. The rapid and accurate identification of this bacterium is of great importance. The aim of this study was to investigate the contamination of raw milk from Tehran cattle farms with S. aureus using culture and real-time PCR methods. For this study, 60 raw milk samples were collected from different cattle farms in Tehran. The samples were first analyzed using the culture method on Baired-Parker agar medium. The colonies with black centers and light areas were used for the coagulase test with rabbit plasma. The real-time PCR technique and the transcriptional regulator gene (Sa0836) specifically developed for S. aureus were then used to confirm the results. Of the 60 raw milk samples, 40 samples (66.66%) were identified as Staphylococcus in the culture medium. Of these, 32 samples (53.33%) were positive in the coagulase test. The results of the molecular tests also showed that S. aureus infection was confirmed in 27 samples (45%). The results of this study show that the contamination of raw milk in Tehran is significant and that the use of culture methods as a gold standard together with molecular techniques such as real-time PCR can help to increase the speed and accuracy of detection of S. aureus in raw milk. This approach can help to improve the quality and safety of dairy products and maintain public health.
1- Górska-Warsewicz H, Rejman K, Laskowski W, Czeczotko M. Milk and dairy products and their nutritional contribution to the average polish diet. Nutrients. 2019 ;11(8):1771.
2- Pal M, Gutama KP, Koliopoulos T. Staphylococcus aureus, an important pathogen of public health and economic importance: A comprehensive review. Journal of Emerging Environmental Technologies and Health Protection. 2021;4(2):17-32.
3- Alembo EA, Tonjo Torka T. Prevalence, Contamination Level, and Associated Factors of Methicillin‐Resistant Staphylococcus aureus in Raw Cow Milk at Selected Districts of Gamo Zone, Southern Ethiopia. Veterinary Medicine International. 2023;2023(1):6238754.
4- González-Martín M, Corbera JA, Suárez-Bonnet A, Tejedor-Junco MT. Virulence factors in coagulase-positive staphylococci of veterinary interest other than Staphylococcus aureus. Veterinary Quarterly. 2020; 40(1):118-31.
5- McAdow M, Missiakas DM, Schneewind O. Staphylococcus aureus secretes coagulase and von Willebrand factor binding protein to modify the coagulation cascade and establish host infections. Journal of innate immunity. 2012;4(2):141-8.
6- Peetermans M, Verhamme P, Vanassche T. Coagulase activity by Staphylococcus aureus: a potential target for therapy? InSeminars in thrombosis and hemostasis 2015 Jun (Vol. 41, No. 04, pp. 433-444). Thieme Medical Publishers.
7- França A, Gaio V, Lopes N, Melo LD. Virulence factors in coagulase-negative staphylococci. Pathogens. 2021;10(2):170.
8- Marsilio F, Di Francesco CE, Di Martino B. Coagulase-positive and coagulase-negative staphylococci animal diseases. InPet-To-Man Travelling Staphylococci 2018 Jan 1 (pp. 43-50). Academic Press.
9- Hussain MA, Shaukat H, Gohar UF. Coagulase-negative staphylococci and their pathogenic role: A focus on nosocomial infections. Applied Biological Research. 2021; 23(2): 113-121
10- Salamandane A, Oliveira J, Coelho M, Ramos B, Cunha MV, Malfeito-Ferreira M, Brito L. Enterotoxin-and antibiotic-resistance-encoding genes are present in both coagulase-positive and coagulase-negative foodborne Staphylococcus strains. Applied Microbiology. 2022; 2(2):367-80.
11- Gajewska J, Chajęcka-Wierzchowska W, Zadernowska A. Occurrence and characteristics of Staphylococcus aureus strains along the production chain of raw milk cheeses in Poland. Molecules. 2022; 27(19):6569.
12- Gebremedhin EZ, Ararso AB, Borana BM, Kelbesa KA, Tadese ND, Marami LM, Sarba EJ. Isolation and identification of Staphylococcus aureus from milk and milk products, associated factors for contamination, and their antibiogram in Holeta, Central Ethiopia. Veterinary Medicine International. 2022; 2022(1):6544705.
13- Kim E, Yang SM, Won JE, Kim DY, Kim DS, Kim HY. Real-time PCR method for the rapid detection and quantification of pathogenic Staphylococcus species based on novel molecular target genes. Foods. 2021; 10(11):2839.
14- Chamchoy T, Okello E, Williams DR, Tonooka K, Glenn K, Maehana K, Gardner IA, Aly SS. Bayesian estimation of sensitivity and specificity of a rapid mastitis test kit, bacterial culture, and PCR for detection of Staphylococcus aureus, Streptococcus species, and coliforms in bovine milk samples. Journal of Dairy Science. 2022;105(7):6240-50.
15- Dibbern AG, Gonçalves JL, Alves BG, Barreiro JR, Anderson K, Santos MV. Real-time polymerase chain reaction for detection and enumeration of Staphylococcus aureus and Streptococccus agalactiae using different milk samplings. Revista Brasileira de Zootecnia. 2023;52:e20210086.
16- Goto M, Takahashi H, Segawa Y, Hayashidani H, Takatori K, Hara-Kudo Y. Real-time PCR method for quantification of Staphylococcus aureus in milk. Journal of food protection. 2007;70(1):90-6.
17- Grace D, Wu F, Havelaar AH. Milk Symposium review: Foodborne diseases from milk and milk products in developing countries—Review of causes and health and economic implications. Journal of dairy science. 2020;103(11):9715-29.
18- Quintana ÁR, Seseña S, Garzón A, Arias R. Factors affecting levels of airborne bacteria in dairy farms: A review. Animals. 2020;10(3):526.
19- Liu D, Lawrence ML, Austin FW. Evaluation of PCR primers from putative transcriptional regulator genes for identification of Staphylococcus aureus. Letters in applied microbiology. 2005;40(1):69-73.
20- Fooladi AI, Tavakoli HR, Naderi A. Detection of enterotoxigenic Staphylococcus aureus isolates in domestic dairy products. Iranian journal of microbiology. 2010; 2(3):137.
21- Hassani S, Hosseini Doust R, Mohebati Mobarez A. Enterotoxin A gene barrier Staphylococcus aureus within traditionally dairy products of Tehran. International Journal of Enteric Pathogen. 2014; 2(4):1-4.
22- Saadat YR, Fooladi AA, Shapouri R, Hosseini MM, Khiabani ZD. Prevalence of enterotoxigenic Staphylococcus aureus in organic milk and cheese in Tabriz, Iran. Iranian journal of microbiology. 2014; 6(5):345.
23- Haghi F, Daneshamooz S, Parsadanians A, Zeighami H. The frequency of Staphylococcus aureus classical enterotoxin genes in raw milk samples in Zanjan, Iran. Journal of Human Environment and Health Promotion. 2019; 5(1):32-5.
24- Shariatifar M, Salavati-Hamedani M, Rezaeian-Doloei R, Rezaeigolestani M, Mohsenzadeh M. Antibiotic resistance and enterotoxin gene profiles of Staphylococcus aureus isolated from raw milk in Iran. Journal of the Hellenic Veterinary Medical Society. 2023; 74(1):5433-40.
25- Fagundes H, Barchesi L, Nader Filho A, Ferreira LM, Oliveira CA. Occurrence of Staphylococcus aureus in raw milk produced in dairy farms in São Paulo state, Brazil. Brazilian Journal of Microbiology. 2010; 41:376-80.
26- Song M, Bai Y, Xu J, Carter MQ, Shi C, Shi X. Genetic diversity and virulence potential of Staphylococcus aureus isolates from raw and processed food commodities in Shanghai. International Journal of Food Microbiology. 2015; 195:1-8.
27- Ágredo-Campos ÁS, Fernández-Silva JA, Ramírez-Vásquez NF. Staphylococcus aureus, Escherichia coli, and Klebsiella spp. prevalence in bulk tank milk of Colombian herds and associated milking practices. Veterinary World. 2023;16(4):869.
28- Aqib AI, Ijaz M, Farooqi SH, Raza A. Dairy Staphylococcus aureus: Epidemiology, drug susceptibilities, drug modulation and preventive measures. London, UK: Intech Open Limited. 2019:57-73.
29- Sharifi A, Sobhani K, Mahmoudi P. A systematic review and meta-analysis revealed a high-level antibiotic resistance of bovine mastitis Staphylococcus aureus in Iran. Research in Veterinary Science. 2023; 161:23-30.
30- Ashraf A, Imran M. Causes, types, etiological agents, prevalence, diagnosis, treatment, prevention, effects on human health and future aspects of bovine mastitis. Animal health research reviews. 2020;21(1):36-49.
