Bacteria of Campylobacter genus are ones of the main zoonotic pathogens causing human and animal diseases. Campylobacter organisms are microaerophiles and, therefore, require low oxygen concentration (3–5%) and high carbon dioxide concentration (3–10%) for their growth. They use amino acids rather than carbons as a source of energy. Classical bacteriological methods for Campylobacter spp. detection are not always successful due to diffi culties in creating optimal conditions for their growth. Therewith, development and implementation of molecular methods for Campylobacter detection and identifi cation are of current importance. Assay for qualitative Campylobacter spp. detection with real-time polymerase chain reaction using CFX-96 thermocycler was optimized. Highly specifi c segment of 16S rRNA gene allowing identifi cation of 6 Campylobacter species: C. jejuni, C. coli, C. lari, C. upsaliensis, C. helveticus и C. hyointestinalis was selected as an amplifi cation target. Optimal magnesium ion concentration (2.5 мМ) and primer annealing temperature (58 °С) were determined. Eighteen reference strains of various bacteria were tested. Only tests of Campylobacter genus strains gave positive results. The method sensitivity was 40 target molecules. The said method was used for testing 76 samples of raw materials of animal origin. Campylobacter spp. genome was detected in 18 samples. Obtained results showed that the optimized variant of real-time polymerase chain reaction based on 16S rRNA gene amplifi cation was a specifi c, sensitive, rapid, reproducible and accurate method for qualitative detection of Campylobacter spp. in samples of raw animal materials.

Staphylococci are one of the causes of food poisoning in many countries of the world. Intoxication occurs due to staphylococcal exotoxins entering the human body. One of the main sources of staphylococcal toxins is milk and dairy products contaminated with pathogenic staphylococci. Staphylococcus aureus has the greatest sanitary and hygienic importance. In 2016–2018 168 samples of ready-to-eat dairy products were tested for Staphylococcus aureus in the Food Safety Laboratory of the FGBI “ARRIAH” in the Republic of Crimea. The tests were performed according to GOST 30347-2016 “Milk and dairy products. Methods of Staphylococcus aureus detection”. Biochemical properties of the recovered isolates were studied using Vitek 2 Compact analyzer. It was established that the following groups of products are contaminated with Staphylococcus aureus to the greatest extent: butter (20%), sour cream (9.09%), curd and curd products (4.55%), pasteurized milk in the consumer packaging (4.35%). The basic biological characteristics of the isolates have been studied and their antimicrobial resistance has been determined. All the isolated Staphylococcus aureus cultures demonstrated a 100% sensitivity to benzylpenicillin, oxacillin, imipenem, ticarcillin, meropenem, ciprofl oxacin, ofl oxacin, gentamicin, amikacin, doxycycline, tetracycline, rifampin, chloramphenicol, cefotaxime, ceftriaxone, trimethoprim and were 100% resistant to enrofl oxacin. Resistance to streptomycin was determined in 28.6% of isolates, and 14.3% of isolates were resistant to vancomycin. Methicillin-resistant staphylococci were not detected among the bacteria.

Salmonella continues to be the primary cause of foodborne intestinal infections in many countries around the world. According to the offi cial data, 47% of the infection outbreaks in the world are associated with salmonellosis, while chicken meat (34%) plays a signifi cant role in the infection transmission to humans through food. Since the early 90’s of the last century, with the massive use of antibiotics, Salmonella strains resistant to a number of antimicrobials began to appear and currently pose a serious public health problem. Resistant strains persistent in animals can be transmitted to humans through the food chain. The paper presents results of studies of morphological, biochemical, serological properties of Salmonella bacteria recovered from animal raw material: beef, pork, poultry meat, tallow, off al derived from broiler chickens and pig slaughter products. In 2018 the FGBI “ARRIAH” Microbiological Laboratory performed 1,204 tests of animal raw material for Salmonella bacteria and recovered 45 Salmonella isolates. Salmonella bacteria were isolated in accordance with GOST 31659-2012 (ISO 6579:2002). Most Salmonella isolates (56%) were recovered from poultry meat. Biological properties of all the studied isolates were quite typical: they formed hydrogen sulfi de, fermented glucose and mannitol with the formation of gas and acid, did not utilize sucrose, lactose and urea; reaction to indole was negative. It was established that the recovered Salmonella isolates belong to serogroups O₇, O₉, O₅, O₄. The frequency of recovering Salmonella group B was 8.9%, group C – 51.1%, group D – 40.0%. Among Salmonella group B, S. derby (4.4%) and S. typhimurium (2.2%) were more common; group C – S. infantis (29.0%), S. virchow (17.8%); group D – S. enteritidis (40.0%). Isolated cases of S. reading (2.2%) and S. oranienburg (4.4%) were observed. All Salmonella isolates recovered from raw material of animal origin demonstrated sensitivity to ciprofl oxacin, chloramphenicol, amoxicillin, amikacin, azithromycin, meropenem, gentamicin, ceftriaxone, kanamycin; were less sensitive to cefotaxime, ampicillin, levofl oxacin and had low sensitivity to nalidixic acid, doxycycline, streptomycin, tetracycline. The phenomenon of multiresistance is characteristic of 44.4% of the isolated Salmonella isolates.

Cattle respiratory diseases are some of the most spread pathologies that can cause economic damage, resulting from fi nancial losses and costs of treatment and diagnostics. One of the major factors contributing to respiratory pathology development is bovine respiratory syncytial infection. The analysis of serological testing, performed by the FGBI “ARRIAH” Reference Laboratory for Cattle Diseases in 2017–2018, showed that respiratory syncytial virus seroprevalence in animals of dairy farms is 60%. Herewith, it was noted that the most susceptible animals to this infection are calves under one year of age. The eff ectiveness of bovine respiratory syncytial infection control measures depends on timely diagnosis; that is why reliable and accurate diagnostic tools are needed, including optimal techniques of virus isolation from pathological material. For successful virus isolation from clinical samples, it is necessary to adhere strictly to optimal parameters of this agent cultivation. This paper presents data on study of bovine respiratory syncytial virus strain AB 1908 cultural properties. The tests performed showed that a continuous bovine turbinate (BT) cell line, continuous bovine fetal trachea (FBT) cell line and continuous bovine calf kidney (RBT) cell line are sensitive for cultivation of this agent and can be used to prepare viral suspension, needed for further research. Virus titre in BT cell culture was 4.33 ± 0.16 – 4.66 ± 0.12 lg TCID50/ cm³, in RBT cell culture – 4.33 ± 0.33 – 4.7 ± 0.36 lg TCID₅₀/cm³ and in FBT cell culture – 4.13 ± 0.20 – 4.78 ± 0.17 lg TCID₅₀/cm³. The following virus cultivation optimal parameters were also determined during this study: the age of the culture for virus inoculation should be 1–2 days and multiplicity of inoculation should be 0.1 TCID₅₀/cell.

Rabies cases are still reported in various mammal species in the Russian Federation. Wild carnivores contacting with domestic and farm animals and transmitting the pathogen to them are the main source and natural reservoirs of rabies. The highest level of rabies incidence in livestock is observed among cattle. Thus, 158 new outbreaks were reported in the Russian Federation in 2017 where the virus was identifi ed in cattle. Rabies specifi c prophylaxis as well as assessment of its effi cacy based on the level of accumulated antirabies virus neutralizing antibodies currently occupy one of the leading positions in the modern system of veterinary and sanitary measures against rabies. The paper presents the study results confi rming effi cacy and safety of three inactivated rabies vaccines based on the “ARRIAH” strain and intended for rabies prevention in cattle. The adsorbed and two emulsion rabies vaccines, formulated with diff erent adjuvants demonstrated innocuity and safety, and induced post-vaccination immunity in all immunized animals at day 21 post inoculation. The mean titers of anti-rabies antibodies were as follows: for inactivated adsorbed vaccine – 4.02 ± 0.76 IU/cm³; for inactivated emulsion vaccines – 16.30 ± 2.03 and 20.73 ± 3.39 IU/cm³. As compared with the adsorbed vaccine, the emulsion vaccines formulated with Montanide ISA 206 and ISA 70 adjuvants induce stronger immunity.

A large number of diff erent groups of livestock diseases in which heredity plays a signifi cant role are currently reported. In particular, more than 500 genetically determined morphological and functional disorders have been detected in cattle; for 150 of them, specifi c mutations are known. The sale of bovine genetic materials is associated with the spread of various diseases caused by mutations that occur in the prominent representatives of breeds. The abundance of lethal mutations in the populations requires a broader application of molecular diagnostic methods for detection of monogenic hereditary diseases. DNA pyrosequencing, being the most convenient technique for the rapid diagnosis of single nucleotide mutations in the bovine genome that are located in the regions with known nucleotide sequence, has a potential for meeting this need. Pyrosequencing-based methods for identifi cation of the most common signifi cant mutations in Holstein, Simmental, Brown Swiss and Aberdeen Angus cattle were developed, validated and approved at the FGBU “VGNKI”. Such mutations are associated with leukocyte adhesion defi ciency, complex vertebral malformation, uridine monophosphate synthetase defi ciency, citrullinemia, spinal muscular atrophy, spinal cord demyelination, Brown Swiss haplotype 2, Weaver syndrome, developmental duplications, α-mannosidosis, dwarfi sm, bovine male subfertility, trombocytopathya, arachnomelia syndrome, hypozincemia-like syndrome. These methods are intended to test semen in the breeding centres, scientifi c laboratories working in the fi eld of biotechnology and animal reproduction, livestock reproduction centres. The use of the proposed genetic tests to detect mutant alleles, as well as reduced use of mutation-bearing animals in stock breeding will allow minimizing the occurrence of inherited diseases and thus improving the gene pool of cattle in the country.

The paper presents data on the study of genetic characteristics of the infl uenza virus A/chicken/ Chelyabinsk/30/2019 H9N2 isolated from pathological material (chicken internal organs) in February 2019 and received from the poultry farm in the Chelyabinsk Oblast. The H9N2 subtype of the isolated virus was identifi ed based on virological analysis. Sequencing of the hemagglutinin gene segment revealed that the amino acid sequence at the cleavage site was RSSR/GLF, which is characteristic of a low virulent avian infl uenza virus. Phylogenetic analysis of the obtained nucleotide sequences of the hemagglutinin gene fragment (1–1539 bp open reading frame) showed that the A/chicken/Chelyabinsk/30/2019 H9N2 isolate belongs to the G1 genetic group of the low virulent infl uenza virus A/H9, the representatives of which are widely spread in the Middle Eastern and Central Asian countries. The complete nucleotide genome sequence of the studied pathogen was determined. The comparative analysis of all genomic segments using the GenBank database revealed a close relationship (over 99%) between the A/chicken/Chelyabinsk/30/2019 H9N2 virus and the A/H9 infl uenza virus isolates circulating in Israel in 2006–2012. According to the analysis of the predicted amino acid sequence of the studied isolate, the positions of some molecular markers that determine the biological properties of the virus have been identifi ed. Most amino acid positions of hemagglutinin (according to H3 subtype sequence numbering) suggest affi nity for the ACA2-3Gal-receptors of avian epithelial cells. Amino acid substitutions were detected at the site within the receptor-binding domain as compared to the A/H9N2 infl uenza virus isolates obtained in Russia in 2018. The primary structure of the A/chicken/Chelyabinsk/30/2019 H9N2 isolate demonstrates a very high level of genetic similarity to the infl uenza virus isolate A/chicken/ Israel/215/2007 H9N2 used as a vaccine strain.

Organizations of all forms of ownership engaged in the production, transportation, procurement and processing of animal products and raw materials, as well as backyards, produce a signifi cant amount of biological waste in the course of their activity. This waste is the source of environmental pollution, it contributes to the maintenance of infectious disease outbreaks and poses a real threat to public and livestock health. Facilities for biological waste rendering and disposal require constant monitoring and supervision, because the improper management of these facilities can lead to disease occurrence and spread with signifi cant environmental, economic and social implications. In order to objectively refl ect the real state of waste rendering and disposal facilities in the Subjects of the Russian Federation and to form a holistic view of the problem in the country, the analysis of data, collected from the veterinary executive authorities of the Subjects of the Russian Federation, were analyzed. Such parameters as quantity, form of ownership, veterinary and sanitary condition, location and availability of animal waste rendering and disposal facilities were considered as of 2018. The study revealed 20,808 animal waste rendering and disposal facilities registered in the country, the majority of them being represented by animal burial sites (including anthrax). In most cases, animal burial sites (including anthrax) do not meet veterinary and sanitary requirements and are unattended. The analysis reveals tension in the fi eld of animal waste management in the Russian Federation.

Major resources for any organization are human resources which condition the company’s performance. The article presents the analysis of quantitative and qualitative indicators of the number, structure and movement of staff of the state veterinary service of the Russian Federation in 2018. All indicators were examined at diff erent organizational levels of the veterinary service: the country’s veterinary service as a whole, at the level of the executive veterinary bodies of Russian subjects (including the state veterinary surveillance divisions), establishments for control and prevention and those for laboratory analysis and diagnosis. The work deals with the staff structure of the country’s state veterinary service categorized by education, length of service, age and gender. To study the movement of staff of the state veterinary service connected to recruitment and retirement of specialists, we calculated and analyzed a number of parameters: employee turnover rate for recruitment, employee turnover rate for leaving, total staff turnover rate, staff replacement rate. In 2018 the Russian state veterinary service comprised around 52,304 veterinary specialists, the majority of them (81%) being employed at establishments for control and prevention. The core of veterinary specialists is represented by employees with a university degree (75%), work experience of more than 10 years (55%), belonging to age group of 36–50 years (43%), female (62%). The results of analysis of human resources of the veterinary service allow us to determine the weak points of personnel policy and work out measures to deal with them if the need arises.