Characterization of the biological properties of bacteriophages Staphylococcus aureus variant bovis
Cattle mastitis is the main cause of economic losses in milk production worldwide, and Staphylococcus aureus is the pathogen that causes it most. Bacteriophages may be an alternative treatment for this disease. In this study, we studied the effect of temperature and pH on the lytic activity of bacteriophages isolated from cows with signs of mastitis. The isolation and production of pure phage lines were performed on an indicator culture of Staphylococcus aureus var. bovis 1491f conventional methods. The following phages were isolated and labeled: Phage SAvB07, Phage SAvB08, Phage SAvB12, Phage SAvB14. To determine the effect of temperature and pH, aliquots after the action of these factors were sown by the double agar method at regular intervals. The study found that phage lytic activity was temperature dependent. Thus, under the influence of temperature 45 °C after 30 minutes of action, it decreased by 3.0–3.4 times for bacteriophages Phage SAvB07, Phage SAvB08, Phage SAvB12 and after one hour was 2.4–12.6%. Phage SAvB14 strain was more resistant to temperature. Its activity decreased by only 67.6% during the analyzed period. With higher temperatures (55–65 °C), the intensity of phage infection decreased significantly, but remained stable. The most resistant to the effects of temperature was Phage SAvB14 – its activity was on average higher by 15.6–33.9% compared with other phages taken in the experiment. The results of our studies on the effect of pH on the reproduction of phages showed that the maximum number of phage virions was observed at pH 6 7. However, the most resistant to acidity was the phage strain Phage SAvB14, compared with other strains taken in the experiment. Therefore, the bacteriophage Phage SAvB14 exhibited the greatest stability and has considerable potential for in vivo use in the treatment of mastitis of cows caused by Staphylococcus aureus.
Gogoi-Tiwari, J., Williams, V., Waryah, C.B., Eto, K.Y., Tau, M., Costantino, P., & Mukkur, T. (2015). Com-parative studies of the immunogenicity and protective potential of biofilm vs planktonic Staphylococcus au-reus vaccine against bovine mastitis using non-invasive mouse mastitis as a model system. Biofoul-ing, 31(7), 543–554. doi: 10.1080/08927014.2015.1074681.
Hanlon, G.W. (2007). Bacteriophages: an appraisal of their role in the treatment of bacterial infections. In-ternational Journal of Antimicrobial Agents, 30(2), 118–128. doi: 10.1016/j.ijantimicag.2007.04.006.
Horiuk, Y.V. (2018). Fagotherapy of cows mastitis as an alternative to antibiotics in the system of obtaining environmentally safe milk. Scientific Messenger of LNU of Veterinary Medicine and Biotechnologies. Se-ries: Veterinary Sciences, 20(88), 42–47. doi: 10.32718/nvlvet8807.
Horiuk, Yu., Kukhtyn, M., Kovalenko, V., Kornienko, L., Horiuk, V., & Liniichuk, N. (2019). Biofilm formation in bovine mastitis pathogens and the effect on them of antimicrobial drugs. Independent Journal of Man-agement & Production, 10(7), 897–910. doi: 10.14807/ijmp.v10i7.1012.
Horiuk, Yu.V., Kukhtyn, M.D., Perkiy, Yu.B., & Horiuk, V.V. (2018). Distribution of main pathogens of masti-tis in cows on dairy farms in the western region of Ukraine. Scientific Messenger of Lviv National Uni-versity of Veterinary Medicine and Biotechnologies, 20(83), 115–119. doi: 10.15421/nvlvet8322.
Jensen, K.C., Hair, B.B., Wienclaw, T.M., Murdock, M.H., Hatch, J.B., Trent, A.T., & Berges, B.K. (2015). Isola-tion and host range of bacteriophage with lytic activi-ty against methicillin-resistant Staphylococcus aureus and potential use as a fomite decontaminant. PLoS One, 10(7), e0131714. doi: 10.1371/journal.pone.0131714.
Karreman, H.J. (2007). Phytotherapy for dairy cows. Veterinary Herbal Medicine. SG Wynn and BJ Foug-ere, ed. Mosby, St. Louis, MO.
Klopatek, S., Callaway, T.R., Wickersham, T., Sheridan, T.G., & Nisbet, D.J. (2018). Bacteriophage Utilization in Animal Hygiene. Bacteriophages: Biology, Tech-nology, Therapy, 1–28. doi: 10.1007/978-3-319-40598-8_30-1.
Kovalenko, V.L., Kovalenko, P.L., Ponomarenko, G.V., Kukhtyn, M.D., Midyk, S.V., Horiuk, Y.V., & Garkav-enko, V.M. (2018). Changes in lipid composition of Escherichia coli and Staphylococcus areus cells under the influence of disinfectants Barez®, Biochlor® and Geocide®. Ukrainian Journal of Ecology, 8(1), 547–550. doi: 10.15421/2018_248.
Kukhtyn, M.D., Horyuk, Y.V., Horyuk, V.V., Yaroshenko, T.Y., Vichko, O.I., & Pokotylo, O.S. (2017). Biotype characterization of Staphylococcus aureus isolated from milk and dairy products of private production in the western regions of Ukraine. Regulatory Mecha-nisms in Biosystems, 8(3), 384–388. doi: 10.15421/021759.
Kukhtyn, M.D., Kovalenko, V.L., Horyuk, Y.V., Horyuk, V.V., & Stravskyy, Y.S. (2016). Bacterial biofilms formation of Cattle mastitis pathogens. Journal for Veterinary Medicine, Biotechnology and Biosafety, 2(4), 30–32. http://jvmbbs.kharkov.ua/archive/2016/ volume2/issue4/pJVMBBS_2016024_030-032.pdf.
Lin, D.M., Koskella, B., & Lin, H.C. (2017). Phage thera-py: an alternative to antibiotics in the age of multi-drug resistance. World journal of gastrointestinal pharmacology and therapeutics, 8(3), 162. doi: 10.4292/wjgpt.v8.i3.162.
Lopetuso, L., Giorgio, M., Saviano, A., Scaldaferri, F., Gasbarrini, A., & Cammarota, G. (2019). Bacteriocins and Bacteriophages: Therapeutic Weapons for Gas-trointestinal Diseases? International Journal of Mo-lecular Sciences, 20(1), 183. doi: 10.3390/ijms20010183.
Mahony, J., McAuliffe, O., Ross, R.P., & van Sinderen, D. (2011). Bacteriophages as biocontrol agents of food pathogens. Current Opinion in Biotechnology, 22(2), 157–163. doi: 10.1016/j.copbio.2010.10.008.
Merabishvili, M., Pirnay, J. P., Verbeken, G., Chanishvili, N., Tediashvili, M., Lashkhi, N., Glonti, T., Krylov, V., Mast, J., Van Parys, L., Lavigne, R., Volckaert, G., Mattheus, W., Verween, G., De Corte, P., Rose, T., Jennes, S., Zizi, M., De Vos, D., & Vaneechoutte, M. (2009). Quality-controlled small-scale production of a well-defined bacteriophage cocktail for use in human clinical trials. PLoS One, 4(3), e4944. doi: 10.1371/journal.pone.0004944.
Moodley, A., Kot, W., Nalgard, S., Jakociune, D., Neve, H., Hansen, L. H., & Vogensen, F.K. (2019). Isolation and characterization of bacteriophages active against methicillin-resistant Staphylococcus pseudintermedi-us. Research in Veterinary Science, 122, 81–85. doi: 10.1016/j.rvsc.2018.11.008.
O’Flaherty S, Ross RP, Flynn J, Meaney WJ, Fitzgerald GF, Coffey A (2005a). Isolation and characterisation of two anti-staphylococcal bacteriophages specific for pathogenic Staphylococcus aureus associated with bovine infections. Letters in Applied Microbiology, 41, 482–486. doi: 10.1111/j.1472-765X.2005.01781.x.
O'Flaherty, S., Coffey, A., Meaney, W. J., Fitzgerald, G. F., & Ross, R.P. (2005b). Inhibition of bacteriophage K proliferation on Staphylococcus aureus in raw bovine milk. Letters in Applied Microbiology, 41, 247–279. doi: 10.1111/j.1472-765X.2005.01762.x.
Rehman, S., Khan, T., & Raza, S. (2016). Isolation of bacteriophage against Staphylococcus aureus causing mastitis. International Journal of Advanced Multidis-ciplinary Research, 3(4), 1393–1395.
Saglam, A. G., Sahin, M., Celik, E., Çelebi, O., Akca, D., & Otlu, S. (2017). The role of staphylococci in subclini-cal mastitis of cows and lytic phage isolation against to Staphylococcus aureus. Veterinary World, 10(12), 1481. doi: 10.14202/vetworld.2017.1481-1485.
Svircev, A., Roach, D., & Castle, A. (2018). Framing the future with bacteriophages in agriculture. Viruses, 10(5), 218. doi: 10.3390/v10050218.
Topuzoglu, B, Bastan, A., & Salar, S. (2015). The effect of long term antibiotic treatment on bacteriological cure and somatic cell count at subclinical mastitis due to Staphylococcus aureu s in lactating dairy cows. Veterinary Journal of Ankara University, 62, 289–294.
Wang, Z., Zheng, P., Ji, W., Fu, Q., Wang, H., Yan, Y., & Sun, J. (2016). SLPW: A virulent bacteriophage targeting methicillin-resistant Staphylococcus aureus in vitro and in vivo. Frontiers in microbiology, 7, 934. doi: 10.3389/fmicb.2016.00934.
Wills, Q. F., Kerrigan, C., & Soothill, J. S. (2005). Experi-mental bacteriophage protection against Staphylo-coccus aureus abscesses in a rabbit model. Antimicro-bial Agents and Chemotherapy, 49(3), 1220–1221. doi: 10.1128/AAC.49.3.1220-1221.2005.
Abstract views: 1 PDF Downloads: 0