Study of the antimicrobial activity of phytoointment for treatment of mechanical damages of dairy tissue

Abstract

Introduction. One of the common pathologies found in gynaecological practice is cracking nipples, which represent the rupture of an external sensitive epidermis due to its mechanical damage. Cracks have different shapes (straight and stellar), as well as different depths (superficial and deep). The nature of cracks is quite diverse: poor quality personal care products or non-compliance with personal hygiene rules, low-quality gels and cosmetics, improper breastfeeding, vitamin A and E deficiency, early use of hormonal contraceptives and other medicines, tanning bed, improperly selected linen and other. Diagnosis is performed only when there are complaints, namely: when the nipple swells, blushing, pain and heaviness in the chest are felt. The treatment is prescribed by the doctor depending on the duration of the disease and its severity. If the disease lasts up to three days, then wound healing, antiseptic and palliative drugs are prescribed. If the disease lasts more than three days, then antibiotic therapy is added to these agents, as there is a secondary infection. In such cases, the breastfeeding of the baby stops.Taking into account the above, the composition of the soft dosage form for use in gynaecology, which has reparative, wound healing action, was developed. Ointment under the conventional name "Phytolan" contains in its composition vegetable oils: amaranth, sea buckthorn, parsley leaves, melaleuca and lanolin anhydrous as the basis. The aim of the research was comparative study of the spectrum and level of antimicrobial activity in vitro ointment "Phytolan", the comparator preparation - the analogue by the action of the cream "Mama care" and specimens containing ointment base - anhydrous lanolin and a separate phytooil in a concentration of 10%. Material&methods.As test strains we took standard strains from the American standard collection of microorganisms: Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 9027, Bacillus subtilis ATCC 6633, Candida albicans ATCC 885-653. Cultivation of cultures was carried out by traditional methods in nutrient media with subsequent confirmation of morphological, cultural and biochemical properties (3). In the course of research, appropriate nutrient media indicated in the national part of the SPHU were used: medium # 1 - in the study of antibacterial activity and medium # 2 - in the study of antifungal activity of the samples. The antimicrobial activity of the drugs was studied in vitro in commonly accepted in microbiological practice method of diffusion in agar in the modification of wells. This method is based on the ability of active substances to diffuse into agar medium, which is pre-sown with the test culture. The molten agar nutrient medium was cooled to 45° C, poured into the bottom layer into Petri dishes in a volume of 10 ml. After hardening of the agar on it were placed six sterile cylinders of stainless steel 10 mm in height and an internal diameter of 8 mm, around which the second layer of medium was poured into a volume of 15 ml, seeded with appropriate cultures of microorganisms. The microbial load was 0,5 units. McFarland turbidity per 1 ml of medium. After clamping the upper layer of agar, the cylinders were removed with sterile tweezers and in the formed wells, the test specimens were administered. Petri dishes were kept for one hour at room temperature, after which they were placed in a thermostat and incubated for 24 hours at a temperature of 37° C with Muller - Hinton agar and 25° C with Sabur agar.The level of antimicrobial activity of the developed ointment, experimental specimens and the comparison preparation was evidenced by the diameter of zones of growth retardation of microorganisms around the well.The research was conducted in six-time repetitions for each test culture. Statistical processing of the results of the study was carried out according to Student's criterion (P <0.5). Results&discussion. According to the results of the research, ointment samples exhibited a moderate level of antimicrobial activity against Staphylococcus aureus culture and practically no activity compared to other cultures. Thus, the growth retardation zone of S. aureus culture around the ointment samples was 13,6 mm, around other cultures 12,1 – 12,2 mm. Other study samples: the comparison drug "Mama care" (analogue by effect) and individually samples with lanolin and amaranth, sea buckthorn, parsley, tea tree almost did not exhibit antimicrobial activity to used test cultures.Despite the known antimicrobial properties of essential oils, in particular tea tree (6), the absence of a pronounced effect in vitro may be due to the technological difficulties of creating an effective antimicrobial concentration in the ointment. The tendency towards the manifestation of antistaphylococcal activity of the ointment may be related to the effect of synergy of essential oils in the composition, in particular the influence of tea tree oil and requires further study of the mechanism of this phenomenon. Conclusion.1. The use of essential oils in the treatment of mastopathy is substantiated. 2. Antimicrobial activity of the developed ointment in relation to S. aureus culture was established and the synergistic effect of essential oils in the composition of the designed drug was revealed. 3. The promise of the application of the developed preparation for the treatment of mechanical damage to the tissues of the mammary gland, complicated by bacterial infections, and the prevention of mastopathy has been proven.