Fagotherapy of cows mastitis as an alternative to antibiotics in the system of obtaining environmentally safe milk

Horiuk, Yulia

doi:10.32718/nvlvet8807

Cited by 9 publications

(8 citation statements)

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“…Проте більше ніж у 50% випадків вони неефективні внаслідок широкого поширення антибіотикорезистентних штамів мікроорганізмів (Kukhtyn et al, 2016;. Формування резистентності до антимікробних препаратів є природним адаптаційним механізмом мікроорганізмів і представляє серйозну медичну та соціально-економічну проблему (Horiuk et al, 2018;Kovalenko et al, 2018). Масштабні дослідження, проведені у країнах Європи та США щодо антибіотикорезистентності збудників маститів, показали, що 57% ізолятів були стійкими хоча б до одного виду антибіотика (Gill et al, 2006).…”

Section: вступunclassified

“…Однак, незважаючи на те, що міжнародні організації пропагують фітотерапію та гомеопатію як альтернативу хімічним речовинам, результати їх ефективності на рівні ферм досить суперечливі. Альтернативними засобами у боротьбі з бактеріальною інфекцією можуть бути використані препарати бактеріофагів (Horiuk, 2018).…”

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Lytic Activity of Staphylococcal Bacteriophage on Different Biotypes of Staphylococcus aureus

Horiuk

2019

SMLNUVMB

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Bacteriophage is a virus that infects a bacterium by injecting a phage genome into a bacterial cytoplasm and uses a host cell as a propagation mechanism. The studied models of phages show a narrow range of hosts. Previous studies on the investigation of lytic activity of staphylococcal bacteriophages were focused on determining the sensitivity of S. aureus isolated from patients from different clinical material and clinically healthy people. However, there is no information as to how refractory are the already described agents against S. aureus ecovars, isolated from animals. The purpose of the work is to study the lytic activity of the agent “Staphylococcal Bacteriophage” in relation to different biotypes of Staphylococcus aureus. Microbiological treatment of samples for isolation of S. aureus was performed using BD Baird-Parker Agar according to standard techniques. To confirm the presence of S. aureus, tests were used for catalase, coagulase, oxidase, for D-mannitol fermentation, DNase production and acetoin. In cultures belonging to S. aureus, the biotype was determined using the scheme: determining the colour of pigment, the presence of beta hemolysis, the activity of coagulase in the bovine plasma, the formation of colonies in a medium with crystal violet. Determination of the range of action of bacteriophages in relation to clinical isolates of microorganisms was carried out by droplet method. The results of determining the lytic activity of staphylococcal bacteriophage in relation to S. aureus isolates of various biological origin showed that the lytic activity of staphylococcal bacteriophage is most active against S. aureus var. hominis. From the studied cultures S. aureus var. hominis full lysis in the course of dripping were found only in 4.8%, and in 42.8% of cultures showed a semiconfluent lysis. 14.3% of cultures S. aureus var. hominis were subjected to weak lytic activity of the bacteriophage. Also, there were detected 4,8% of cultures of this biotype, which were resistant to staphylococcal bacteriophage. In the study of lytic activity of staphylococcal bacteriophage up to 35 cultures S. aureus var. bovis, the manifestation of the lytic action of only one culture is established. Moreover, the level of lysis was estimated at “+/–”, that is, they showed less than 20 phage colonies. At the same time, the studied by us staphylococcal bacteriophage did not show the lytic action on S. aureus var. avium and S. aureus var. canis. Therefore, conducted studies have shown that the lytic activity of the agent “Staphylococcal Bacteriophage” is directed mainly to S. aureus var. hominis, and practically does not work on other biotypes. In consideration of the apparent lack of activity of phage agents in relation to the studied biotypes, attention should be focused on the specificity of phages, not only within the species of bacteria, but also within their biotypes.

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Lytic Activity of Staphylococcal Bacteriophage on Different Biotypes of Staphylococcus aureus

Horiuk

2019

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“…Актуальність теми. Бактеріофаги (віруси, які заражають бактерії) дуже поширені в навколишньому середовищі і можуть бути джерелом недорогих протимікробних препаратів (Kukhtyn et al, 2017;Horiuk, 2018;Altamirano & Barr, 2019). Терапевтичний і протиепідемічний потенціали бактеріофагів визначаються використанням тільки літичних фагів і ступенем їх літичної активності у відношенні до збудників інфекції (Fortuna et al, 2019).…”

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Effect of Temperature on the lytic activity of Bacteriophage PHAGE SAVB14, specific for STAPHYLOCOCCUS AUREUS VARIANT BOVIS

Horiuk¹,

Kukhtyn²,

Horiuk³

et al. 2019

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Therapeutic effect in case of phagotherapy is determined by the degree of lytic activity of bacteriophages in relation to contagions. Various factors influence the lytic activity of phages including temperature, pH, shelf-life, etc. Temperature plays a considerable role in attachment, penetration, reproduction and duration of latent period. The purpose of this research was to learn the influence of different temperature on lytic activity of bacteriophage Phage SAvB14 which is active in relation to Staphylococcus aureus variant bovis. For the increase of bacteriophage titre Phage SAvB14 the strain of gold Staphylococcus aureus var. bovis 1491f (Certificate for the strain-#736, from 05.03.2019) was used which was abstracted from the sample of the secretion of mammary gland of the patient with subclinical form of mastitis. For determination of temperature influence (0, 4, 8 and 45, 55, 65 °C) aliquots were sown by a double agar in definite intervals by the generally accepted methods. The cups were incubated at temperature 37 °C during 18-24 hours. Research was repeated three times. According to the results of research it was determined that warming up of test tubes with bacteriophages at temperature 45 ºС reduced the activity of phages in 30 minutes of influence in 1,7 times. Temperature 55 and 65 ºС had more harmful effect, during the influence for the first 30 minutes the amount of active phages diminished in 2,5 and 5,3 times, and for 60 minutes destroyed the bacteriophage Phage SAvB14 by 89,9 and 93,3% respectively. The obtained results testify the thermolability of Phage SAvB14 which is an important factor in making phage preparation. The most optimum terms for storage of Phage SAvB14 are temperature regime within the limits of 4 and 8 °C. In case of such temperature lytic activity diminished only in 1,5-1,7 times, while in case of 0 °C in 2,4 times compared to the initial amount of bacteriophages.

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“…Recently, many scientists have suggested that, for environmental and physiological reasons, bacteriophages are likely to be more effective than antibiotics in the destruction of bacteria in biofilm (Dias et al, 2013;Horiuk, 2018;Milh et al, 2019). The effect of phages on biofilms includes an initial stage of bacterial adsorption, followed by bacterial infection.…”

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confidence: 99%

Influence of staphylococcal Phage SAvB14 on biofilms, formed by Staphylococcus aureus variant bovis

Horiuk

Kukhtyn

Stravskyy

et al. 2019

Regul. Mech. Biosyst.

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The use of bacteriophages for the treatment of chronic inflammatory processes has proved to be relevant, especially during isolation of antibiotic-resistant pathogens formed in biofilms. The article presents the results of research on the influence of Phage SAvB14 on young and mature biofilms formed by Staphylococcus aureus variant bovis. In the experiments we used cultures of S. aureus and a specific Phage SAvB14 isolated from the secretion of the mammary gland of cows suffering from chronic mastitis. In the study of the influence of bacteriophage on formed biofilms we determined the optical density of the dye solution that was washed from the biofilm photometrically on a spectrophotometer PE-5400UV (Ecroskhim, Russia) and the number of staphylococcal cells in the biofilm after the action of the bacteriophage on 24-hour and 72-hour biofilms by a ten-fold dilution on beef-extract agar. It was determined that under the influence of the bacteriophage on young 24-hour biofilms of S. aureus var. bovis, the optical density of the dye solution from biofilm increased within 4 hours up to 10% and the number of microbial cells increased by 1.8 times. After 32 hours of bacteriophage action, the optical density of the dye solution decreased on average by 34% compared to the initial density and the number of S. aureus cells in the biofilm decreased by 30 times. This indicates that microbial cells of young biofilms are not subject to complete lysis during the action of even this specific bacteriophage. Degradation of 77.5% of biofilm under the influence of the bacteriophage was observed on mature 72-hour biofilm within 32 hours at 37 °C. At the same time, viable cells of S. aureus were not isolated from the biofilm. This indicates the high lytic activity of the bacteriophage against mature biofilm bacteria and the possibility of its use in chronic staphylococcal infections caused by S. aureus var. bovis. Thus, the obtained data indicate that when mature 72-hour biofilms are exposed to the researched bacteriophage, their degradation is more intense compared with the young 24-hour biofilms, and the amount of destroyed biofilm was on average 2 times higher. This suggests that the use of specific staphylococcal Phage SAvB14 isolated by us for the destruction of biofilm, formed by S. aureus var. bovis, is promising.

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Fagotherapy of cows mastitis as an alternative to antibiotics in the system of obtaining environmentally safe milk

Cited by 9 publications

References 26 publications

Lytic Activity of Staphylococcal Bacteriophage on Different Biotypes of Staphylococcus aureus

Lytic Activity of Staphylococcal Bacteriophage on Different Biotypes of Staphylococcus aureus

Effect of Temperature on the lytic activity of Bacteriophage PHAGE SAVB14, specific for STAPHYLOCOCCUS AUREUS VARIANT BOVIS

Influence of staphylococcal Phage SAvB14 on biofilms, formed by Staphylococcus aureus variant bovis

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