Study of the Inhibitory Effect of Probiotics on Staphylococcus Aureus Bacteria Isolated from the Milk of Cows Suffering from Mastitis
DOI:
https://doi.org/10.48165/ijapm.2026.42.01.07Keywords:
conducted, isolate, Staphylococcus, aureusAbstract
This study was conducted to isolate and identify Staphylococcus aureus as one of the main causative agents of bovine mastitis, and to evaluate the inhibitory effect of lactic acid bacteria against the isolated strains. In addition, the antibiotic susceptibility of the isolates was determined using the disk diffusion method A total of 100 milk samples were collected from cows in different areas of Baghdad Governorate, including 40 samples from cows with acute mastitis 30 samples from cows with subclinical mastitis, and 30 samples from apparently healthy cows. Milk samples were cultured on blood agar followed by mannitol salt agar, and the isolates were identified based on microscopic, cultural, and biochemical characteristics The results showed that 10 isolates (10%) were identified as Staphylococcus aureus ,Sixty milk samples were negative by the California Mastitis Test, while no bacterial growth was observed in samples collected from healthy cows. Molecular detection revealed that 10 isolates (10%) were positive for the 16S rRNA gene, whereas 6 isolates (6%) were positive for the nuc gene Furthermore, all tested lactic acid bacteria isolates (Bifidobacterium bifidum, Lactobacillus acidophilus, and Lactobacillus rhamnosus) exhibited a clear inhibitory effect against Staphylococcus aureus at a concentration of 500 µL. However, no inhibitory activity was observed at lower concentrations (250 and 100 µL).
References
Abdullah, R., & Ahmed, R. (2023). Probiotics effect on Gram-positive and Gram-negative bacteria that cause different clinical infections. Journal of Biotechnology Research Center, 19(2). https://www.jobrc.org/index.php/jobrc/article/view/780
Al-Kafaji, Z. M. (1992). Influence of various conditions on inhibitory effect of intestinal Lactobacillus against intestinal bacteria causing diarrhea. Iraqi Journal of Science, 33(1), 18–26.
Abd El-Rahman, H. A., Hasan, A. F., Alyasiri, T., El-Wahsh, H. M., Althubyani, S. A., Basyony, M. A., & Mahmod, D. H. (2024). Co-treatment with cranberry and vitamin C mitigates reproductive toxicities induced by phenobarbital in male rats. Cellular Physiology and Biochemistry, 58, 722–738.
Al-Harbi, A. H., et al. (2021). Prevalence and antimicrobial resistance of Staphylococcus aureus isolated from bovine mastitis in dairy farms. Veterinary World, 14(6), 1615–1621. https://doi.org/10.14202/vetworld.2021.1615-1621
Al-Qassab, A.-J. O., Al-Khafaji, Z. M., & Mahmoud, Z. (1992). The effect of different conditions on the inhibitory activity of enteric lactobacilli against enteric bacteria causing diarrhea. Iraqi Journal of Agricultural Sciences, 23(1), 34–40.
Al-Ugailiity, D. N. (2013). Bacteriological and genetic studies on oxacillin-resistant Staphylococcus aureus isolated from some hospitals in Baghdad (Doctoral dissertation). Al-Nahrain University.
Becker, K., et al. (2020). PCR detection of the nuc gene for confirmation of Staphylococcus aureus. Journal of Clinical Microbiology, 58(2), e01234-19. https://doi.org/10.1128/JCM.01234-19
Boestroo, M. H., Wit, J. C., Kuster, S. J. M., & Rombouts, F. M. (1993). Inhibition of the growth of yeasts in fermented salads. International Journal of Food Microbiology, 17, 311–320.
Capurro, A., Aspán, A., Unnerstad, H. E., Waller, K. P., & Artursson, K. (2010). Identification of potential sources of Staphylococcus aureus in herds with mastitis problems. Journal of Dairy Science, 93(1), 180–191.
Carter, G. R., & Cole, J. R., Jr. (1990). Diagnostic procedures in veterinary bacteriology and microbiology (5th ed.). Academic Press.
Eberhart, R. J., Harmon, R. J., Jasper, D. E., Natzke, R. P., Nickerson, S. C., Reneau, J. K., & Spencer, S. B. (1987). Current concepts of bovine mastitis. National Mastitis Council.
Fehlings, K., Deneke, J., & Wittowski, G. (1997). Prophylactic measures against infection in dairy cattle—Necessity of hygiene management. Deutsche Tierärztliche Wochenschrift, 104(8), 306–312.
Aljeboury, G. H., Risan, M. H., & Algafari, R. N. (2019). Effect of probiotic extracted from Lactobacillus sp. antagonist Helicobacter pylori of human stomach ulcer. Journal of Global Pharma Technology, 11(5 Suppl.), 731–737.
Aljeboury, G. H., & Mahmoud, S. N. (2020). Evaluation of antagonistic potential of Lactobacillus isolates against phytopathogenic fungi and pathogenic bacteria in vitro. Systematic Reviews in Pharmacy, 11(12).
Freeman, B. A. (1985). Barrow’s textbook of microbiology (22nd ed.). Igaku-Shoin/Saunders International.
Hasan, N. S., & Hoshyar, D. F. (2019). Detection of enterotoxigenic Staphylococcus aureus strains in raw milk of cows reared in Erbil Province, Iraq. Zanco Journal of Pure and Applied Sciences, 31(4), 50–60.
Kirby, W. M., Bauer, A. W., Sherris, J. C., & Turk, M. (1966). Antibiotic susceptibility testing by standardized single-disc method. American Journal of Clinical Pathology, 45, 493–496.
Kourkoutas, Y., Bosnea, L., Taboukos, S., Lambrou, D., & Kanellaki, M. (2006). Probiotic cheese production using Lactobacillus casei cells immobilized on fruit pieces. Journal of Dairy Science, 89, 1439–1451.
Lewus, C. B., Kaiser, A., & Montville, T. J. (1991). Inhibition of food-borne bacterial pathogens by bacteriocins from lactic acid bacteria isolated from meat. Applied and Environmental Microbiology, 57, 1683–1688.
Obaid, R. M., Yaseen, F. T., Kadhim, N. K., Salim, D. H., Sabar, Z. T., Abd, D. S., & Hasan, A. F. (2025). Changes in the level of lipid profile in diabetes mellitus in patient samples. Journal of Bioscience and Applied Research, 11(1), 331–336.
Malvisi, M., et al. (2020). Isolation and analysis of milk microbiota in dairy cows with clinical mastitis. Microorganisms, 8(5), 731. https://doi.org/10.3390/microorganisms8050731
Mishra, C., & Lambert, J. C. (1996). Production of antimicrobial substances by probiotics. Asia Pacific Journal of Clinical Nutrition, 5, 20–24.
Mojgani, N., & Ashtiani, M. B. (2006). In vitro inhibition of mastitis pathogens by bacteriocin RN86 produced by an indigenous Lactobacillus casei isolate. Online Journal of Veterinary Research, 10(1), 61–67.
Mokoena, M. P. (2020). Lactic acid bacteria and their bacteriocins: Classification, biosynthesis and applications against uropathogens: A mini-review. Molecules, 25(23), 5561. https://doi.org/10.3390/molecules25235561
National Committee for Clinical Laboratory Standards. (2003). Performance standards for antimicrobial disk susceptibility tests (M2-A8, 8th ed.). NCCLS.
Nunes, S. F., Bexiga, R., Cavaco, L. M., & Vilela, C. L. (2007). Antimicrobial susceptibility of Portuguese isolates of Staphylococcus aureus and Staphylococcus epidermidis in subclinical bovine mastitis. Journal of Dairy Science, 90(7), 3242–3246.
Omran, Z. H., Hussen, A. K., Yahya, A. M., Mohammed, D. A., Rawdhan, H. A., Ahmed, H. M., Sattar, R. J., Hussen, Y. A., Jasm, B. H., & Sadoon, N. M. (2024). Copper nanoparticles against Staphylococcus aureus and Escherichia coli. Journal of Nanostructures, 14(3), 780–788.
Salih, A. M., & Mahdi, M. S. (2025). Antibacterial activity of probiotic Lactobacillus acidophilus against antibiotic-resistant pathogens isolated from skin infection. Journal of Biotechnology Research Center, 19(1), 84–91. https://doi.org/10.24126/jobrc.2025.19.1.906
Sambrook, J., Fritsch, E. F., & Maniatis, T. (1989). Molecular cloning: A laboratory manual (2nd ed.). Cold Spring Harbor Laboratory Press.
Sayhood, M. H., Mohammed, A. L., Abdulhameed, M. F., & Jori, M. M. (2022). Classical and molecular identification of Staphylococcus aureus isolated from cattle wounds with myiasis in Basrah Governorate, Iraq. Iraqi Journal of Veterinary Sciences, 36(3), 641–646.
Schlegelova, J., Babak, V., Klimova, E., Lukasova, J., Narratilova, P., Sustackova, A., Sediva, I., & Rysanek, D. (2002). Prevalence of resistance to antimicrobial drugs in selected microbial species isolated from bulk milk samples. Journal of Veterinary Medicine B, 49, 216–225.
Schneider, M., Valle, M., Woehrle, F., & Boisrame, B. (2004). Pharmacokinetics of marbofloxacin in lactating cows after repeated intramuscular administrations and pharmacodynamics against mastitis isolates. Journal of Dairy Science, 87(1), 202–211.
Schultz, P. J., Smith, K. I., Hogan, J. S., & Love, B. C. (2004). Antimicrobial susceptibility of mastitis pathogens from first lactation and older cows. Journal of Veterinary Microbiology, 102(1–2), 33–42.
Silva, M., Jacobus, N. V., Deneke, C., & Gorbach, S. L. (1987). Antibacterial substances from a human Lactobacillus strain. Antimicrobial Agents and Chemotherapy, 31(8), 1231–1233.
Thornsberry, C., Marler, J. K., Watts, J. L., & Yancy, R. J. (1993). Activity of pirlimycin against pathogens from cows with mastitis and recommendations for disk diffusion test. Antimicrobial Agents and Chemotherapy, 37(5), 1122–1126.
Tokue, Y., Shoji, S., Satoh, K., Watanabe, A., & Motomiya, M. (1992). Detection of methicillin-resistant Staphylococcus aureus using polymerase chain reaction amplification. Tohoku Journal of Experimental Medicine, 163, 31–37.
Tollersrud, T. (2001). Staphylococcus aureus mastitis: Bacterial characteristics and host immune response (Doctoral dissertation). National Veterinary Institute.
Tortora, G. J., Funke, B. R., & Case, C. L. (1986). Microbiology: An introduction (2nd ed.). Benjamin/Cummings.
Vignolo, G. M., Suriani, F., Halgado, A. P. R., & Oliver, G. (1993). Antibacterial activity of Lactobacillus strains isolated from dry fermented sausages. Journal of Applied Bacteriology, 75, 344–349.
Wongboot, W., Chomvarin, C., Engchanil, C., & Chaimanee, P. (2013). Multiplex PCR for detection of superantigenic toxin genes in methicillin-sensitive and methicillin-resistant Staphylococcus aureus isolated from patients and carriers in a hospital in northeast Thailand. Southeast Asian Journal of Tropical Medicine and Public Health, 44(4), 660–671.
Yaseen, T. F., & Al-Jumaily, K. M. R. (2025). The impact of interleukin-21 and 23 serum level and gene expression in celiac disease among Iraqi patients. Asian Journal of Dairy and Food Research, 44(2), 234–239. https://doi.org/10.18805/ajdfr.DRF-476
