Scientific Journal of KubSAU

The work was done in the research laboratory of the department of Biotechnology, Biochemistry and Biophysics of Kuban State Agrarian University, the aim of which was to select the most appropriate nutrient medium for maximum growth of lactic acid microorganism growth. The object of the study was its own microflora of gastrointestinal tract of quails ‒ Lactobacillus agilis, Lactobacillus intermedius and Lactobacillus salivarius. As a nutrient substrate there were used 4 of the nutrient media: the medium for lactic acid bacteria (city Uglich), the melasse-autolysis medium, the glucose-peptone medium and the MRS. During the cultivation of the microflora of the variable parameters were the time and temperature of cultivation. During the cultivation of microflora the time and the temperature of growing were the variable parameters. In the process of growing there was carried out the analysis of dynamics of consumption of reducing substances and titers of microorganisms. According to the results of growing of microbial cultures there was revealed the active consumption of carbon substrate in used variants of nutrient media, and it was found that there was observed the maximum of cells to 24 h of cultivation. On the basis of carried out results of cultivation on different media and at different parameters there was determined that the most titer of cells was reached to 24 h at the temperature 38,0 °С on the melasse-autolysis medium. So, the melasse-autolysis medium can be recommended in production conditions as the cheaper substrate at the further development of biological preparations for poultry breeding

Carotenoid preparations are quite widely used in a health care, but in the veterinary and animal husbandry, their use is limited. In terms of livestock farms of the Krasnodar region and the Rostov region at the biochemical monitoring studies of the cattle blood the lack of carotene in the serum is revealed from 45 to 100 % of the samples. To study the biological function of carotenoid we used preparations, which are the oily solutions for injections: β-carotene (manufactured by FS 42-3867, which is produced from the biomass culture of the fungus Blakeslea trispora) and lycopene, emitted from plants. The cows in the test groups during the dry period and within two weeks after birth got carotenoid preparation subcutaneously at a dose of 10 ml per animal with an interval of 7 days. As a result, it was found out that the application of carotenoids has the most favorable conditions for the flow of labor and the postnatal period, which are manifested in the decrease in obstetric and gynecological diseases in cows and reduce the period of infertility. In the experimental groups the levels of vitamin A and carotene1,3 times higher (1,26 ± 0,14 μmol/L vs. 0,97 ± 0,09 μmol/L ) 1,8 times (0,34 ± 0 03 mg /% vs. 0.61 ± 0.11 mg /%) respectively in comparison with the parameters of the intact animals at a high level of confidence (P < 0,05). Phagocytic number and percentage of digestion of neutrophils increased by 8,5 %, the difference in the number of B-cells reached 9,1%, T-cells – 6,7%. In a humoral immunity found a significant increase in Ig A. After comparison of the level of lysozyme activity of blood serum was observed its reduction in all animals immediately after birth, but in the experimental groups this process was less pronounced. The use of carotene preparations helps to reduce the concentration of lipid peroxidation products in the body of cows (diene conjugates - by 16,6 %, ketodienes - by 35,7 %, malondialdehyde – by 11,3 %), which creates favorable conditions for the flow of the metabolic processes associated with ensuring the normal childbirth and post-partum involution of the genitals. In the group of cows, receiving the carotenoid lycopene, the dyspepsia morbidity of the newborn calves compared to a control decreased by 26,7 %, bronchopneumonia - by 13,3 %

The revolutionary discoveries of recent years in immunology has proved that protection from infections and biological aggression leading role played by innate immunity, which leads to the launch and subsequent work of specific adaptive immunity. Today, it is well-known that almost any pathology of the animal affects the immune system and creates an immune imbalance, leading to development of chronic pathological process, frequent relapses and to the development of different kind of complications. Secondary immunodeficiency states are widely distributed and differ in many etiological factors age, nutritional, medicinal, infectious and neoplastic nature, which leads to increased susceptibility of animal to various раthogenic factors, the weakening of the body. Awareness of the role of immunological disorders in the pathogenesis of a number of diseases has led to the need use in veterinary medicines that are aimed to influence on the immune system of the animal at the known molecular mechanism, to combine safety with high clinical efficacy. In this respect, a promising immunomodulators of microbial origin, due to the knowledge of the recognition mechanisms of the innate immune system such as bacteria as a whole, separate structural units, including mini-optimal biologically active fragments. This review is dedicated to one of the immunomodulator of microbial origin – which is an analog of biologically active fragment of muramyldipeptide of bacterial cell walls ‒ glucosaminylmuramildipeptide (GMDP), which is the active substance of registrated in Russia drug with the trade name Glicopin for use in veterinary practice. The drug showed high efficiency when used in cats, dogs, rodents, ferrets with infections of bacterial and viral etiology; cancer; with the aim of improving the natural resistance of the organism of young and old animals; enhance the effectiveness of vaccination in immunocompromised animals; postvaccine reduce complications; reduce stress financial position, such as during transportation and various veterinary treatments; injuries and on-line intervention (for stimulation of repair processes and tissue regeneration); prevention of immunodeficiency in newborn animals; the reduction of postpartum involutional processes in the uterus. Analysis of literature data allows us to consider an important outcome of immunologiс discoveries of recent years is the change of views on the question of the treatment of infectious diseases and the role of drugs as activators of innate immunity, among which an important role is played by a synthetic analogue of muramyldipeptide (GMDP) is a drug with a known molecular mechanism of action

The work was carried out in the scientific-research laboratory of the chair of Biotechnology, Biochemistry and Biophysics of Kuban SAU, the aim of which is the search of optimal conditions of obtaining the hydrolyzates of lactic acid bacteria by thermoacid and enzymatic ways. The pure cultures of microflora of gastrointestinal tract of quails − Lactobacillus agilis, Lactobacillus intermedius and Lactobacillus salivarius were the object of researches. Under conducting the thermal acid hydrolysis there were used the same correlations of bacterial mass and water as well as procedures of clearing of hydrolyzates. The variable conditions were рН, temperature and time of hydrolysis. When carrying out the enzymatic hydrolysis in this study there was used the enzyme from the group of muramidase lysozyme. The amount of enzymes and time of carried out hydrolysis were the variable conditions under obtaining the enzymatic hydrolyzates. In the process of researches there were studied: qualitative content of total protein, peptides with М.м. < 1500, protein and peptides with M.m. > 1500, aminoacids and as well as GMDP (glucosaminilmuramilpentapeptide). On the basis of carried out experiments there was stated that according to all components of biologically active substances the thermal acid way is more effective than enzymatic. The maximum accumulation of BAA in hydrolyzates by different ways was revealed at destruction of cells Lb. intermedius. Content of GMDP in thermal acid hydrolyzate was higher than in enzymatic on 0,028 g/100 ml (51,8 %)

In the review, the stages of designing therapeutic cocktails of T4 type bacteriophages based on works by Harald Brüssow from Nestlé S.A. (Switzerland) are considered. The main stages of this process are identified: analysis of existing cocktails, selection of phages; creating a collection; cultivation of the host bacterium, multiplication of viruses; purification of the preparation; contamination testing; preservation, stabilization and storage; preclinical and clinical trials. H. Brüssow first studied the Russian drug " Coli-Proteus bacteriophage " of "Microgen" with the help of metagenomic analysis, electron microscopy and conducted its clinical studies. Prof. Brüssow considered the advantages of T4 bacteriophages for the treatment of Escherichia coli infections. Researchers studied methods of cultivation in Erlenmeyer flasks, in a bioreactor, in disposable sack cultivators for the propagation of viruses. For its purification the chromatography, centrifugation, filtration and polyethylene glycol precipitation were studied. To quickly check the contamination of phage cocktails, a mass spectrometry method is proposed. Researchers considered basic strategies, such as lyophilization, spray drying, the formation of microcrystals and microspheres to stabilize the preparations. They also reviewed the results of clinical trials of phage cocktails. We have listed the problems of selecting T4 bacteriophages from the point of view of modern knowledge. H. Brüssow and his colleagues carried out an interesting work on the construction of phage cocktails based on T4 type bacteriophages, and also revealed the problems of the current state of phage therapy

A method of obtaining insertion mutants for the hoc gene, which encodes for the main phage antigen, was developed on the model of bacteriophage T4. This gene was cloned in the plasmid pBSL0+ and was disrupted by insertion of foreign DNA. The phage mutants were obtained by in vivo phage-plasmid recombination. The construction of insertion bacteriophage mutants was carried out in two stages. The resulting mutants on this procedure could be grown on wild-type E. coli strains, which is convenient for the production and use of these phages in therapy. The mutants obtained had reduced antigenicity. At the same time, the yield of the mutant strains was high when they were grown on the non-suppressor E. coli laboratory strains. A number of stages of purification of the bacteriophage mutants obtained were performed. Preparations have been studied by transmission electron microscopy and mass spectrometry. By several periodic cultivations of the mutant bacteriophages, it was shown that mutations of this type are stably maintained during more than 50 generations. T4 related bacteriophages of the family Myoviridae, for example, T-even, have the significant homology amongst their genomes, which makes possible to produce similar mutants. Thus, our method was developed to obtain mutants with reduced antigenicity which can be used for both the treatment of systemic infections, and diarrhea in the case, when, bacteriophages penetrate into the bloodstream. Such phages can be used in medicine and veterinary. The reported study was partially supported by RFBR, research projects No. 13-04-00991, 16-44-230855

We have studied biochemical changes in the body of cattle, sick with nodular dermatitis in the initial stage of the clinical manifestation of the disease. It is proved that in case of disease in animals there is a significant increase in the level of creatinine up to 295.5 mg%. The excess of the norm by 82.1%. In treatment, this indicator was significantly reduced (189.2 mg%), but did not reach the upper limits of the norm (162.4 mg%). In the blood serum of patients, the carotene content was reduced to 0.09 mg%, which was restored to normal three days after the start of treatment (1.0 mg%). The glucose level at the beginning of the disease was 18% below normal and reached the lower limit of the norm 8 hours after the start of treatment. When studying the activity of CKK, it was found that the activity of the enzyme in animals before the introduction of a 5% solution of sodium hydrogencarbonate is 82% higher than the upper limit of the norm. After the treatment, this indicator returned to normal. The inorganic phosphorus index did not reach the physiological norm (2.5 mmol / l) before and after treatment, and remained 11.6% higher (2.9 mmol / l), which indicates a violation of mineral metabolism in sick animals. The level of reserve alkalinity began to recover after 8 hours of observation and reached a minimum limit of 48.91 vol. % CO2. During the entire period of the experimental studies, before and after the administration of the drug, there were no significant changes in the biochemical parameters of total protein, calcium, vitamin E and urea. The nature of the established changes varied within the limits of physiological norms. The use of a 5% solution of sodium hydrogen carbonate in nodular dermatitis increases the effectiveness of symptomatic treatment, reduces the likelihood of complications and stimulates the production of specific antibodies against viruses and bacteria. We have recommended intravenous injection of 5% sodium hydrogencarbonate solution to sick animals at the rate of 1 ml per 1 kg of live weight of the animal three times with an interval of 3 days

Pasteurella multocida is an important respiratory pathogen of cattle. OmpH Protein is a major protective antigen of bacteria has been well studied in avian strains. In the literature there are no data available for the study of a variety of sequence of this protein among isolates with cattle respiratory pathology. There have been described several genes associated with the virulence of the bacterium in respiratory disease of cattle, but none of the authors compared the frequency of detection of these genes with the pathogenicity for laboratory animals. The aim of our study was the development of allele-specific PCR to determine Omph-types of Pasteurella multocida and the search for new approaches to assess the pathogenicity of isolates of bacteria. Total amount of 83 isolates allocated from the lungs of calves with respiratory pathology was investigated. All isolates belonged to groups A or D (isolates 63 and 20, respectively). Among isolates of capsular serogroup A we revealed 6 types, most propagation types were A1 and A2. All isolates of capsular serogroup D were one omph- type. In 16 out of 23 farms there were identified isolates of only one omph-type, 4 - 2 types, 3 - three types. The frequency of gene hgbb - hemoglobin binding protein correlated with pathogenicity of isolates for white mice. The developed allele-specific PCR along with hgbb gene detection can be used for screening and studying the properties of antigen and circulating pathogenic isolates and selecting a candidate vaccine strains

In this work, we have found shortened particle forms of the filamentous bacteriophage Pf, when the lysis of the Pseudomonas aeruginosa PAO1 strain was induced by extracts of bison feces to study the intestinal bacteriophages of this animal. The sizes of the shortened particles from 250 to 950 nm and their thickness 17 nm were determined. The particles had a slightly pointed one end and sometimes a flag on the other extended end. The particles were formed in the process lysis of this strain by other bacteriophages. These lysates also contained a high concentration of vesicles from the pseudomonad membrane. The article discusses the mechanisms of prophage induction in the Pseudomonas aeruginosa PAO1 strain due to phage-phage induction and substances of the investigated extract

The article reflects information characterizing the significance of mastitis and endometritis in the indicator of premature disposal of cows from herds of large farms in the western regions of the Krasnodar region: an average of 30% of the total culling of cows. The effectiveness of veterinary measures in Pravoberezhny JSC of the Temryuk district for mastitis and endometritis in cows was assessed: 81-95%; fertilization index value ‒ 1.8; the duration of the service period exceeds the requirements by 20 days. The studies the nature of the influence of the drugs called Sepranol and Uteroton on the calving process and the occurrence of postpartum complications in cows from the herd, which was formed with violation of breeding principles. The use of the drugs Uteroton and Sepranol contributed to a 50% decrease in the calving index in calving cows by 50% compared with the results of the combined use of estrophan and oxytocin. The use of Uteroton reduced the damage from the lack of milk by 87880 rubles, and the use of Sepranol - by 171164 rubles. We have given practical recommendation on the use of Uteroton and Sepranol for veterinary effects on the body of cows in the early postpartum period in order to reduce the occurrence of complications