More than 30,000 samples of blood serum from domestic, wild and synanthropic birds from 50 regions of the Russian Federation were submitted to the FGBI “ARRIAH” (Vladimir) Reference Laboratory for Avian Viral Diseases to be tested for avian influenza and Newcastle disease within the framework of monitoring activities conducted by the Rosselkhoznadzor in 2019. As a result of the laboratory diagnosis, antibodies to type A influenza virus were detected in vaccinated chickens from two poultry farms in the Perm and Primorsky Krais (A/N9). The detected antibodies were specific to the haemagglutinin subtype of the vaccine antigen. As for the backyards in the RF Subjects, where scheduled vaccination against avian influenza A/H5 is carried out, a low level of immunity was seen in the Rostov and Astrakhan Oblasts (35 and 44%, respectively) while a high level of immunity was observed in the Republic of Altai, Krasnodar Krai, the Chechen Republic and the Primorsky Krai (69, 78, 80 and 88%, respectively). High seroprevalence of Newcastle disease virus in adult poultry in indoor holdings was associated with mass vaccination against the disease. In broiler chickens, post-vaccination antibodies were observed, on average, in 42% of the studied blood serum samples. Antibodies to the Newcastle disease virus were detected in 39% of samples from backyard chickens. Seroprevalence in wild and synanthropic birds was high. The obtained results suggest that the risk of introduction and spread of avian influenza and Newcastle disease in industrial poultry farms and in backyards remains.

The results of the scientific expedition to Tere Khol and Uvs Nuur Lakes in the Republic of Tyva with the purpose of active monitoring of highly dangerous diseases in wild migratory waterfowl and epidemic analysis of these biotope water areas are presented in the paper. The Uvs Nuur Lake is a kind of an indicator for avian influenza introduction to the Russian Federation, because this is the resting and nesting area for many migratory wild birds during the period of mass migrations from Central and South-East Asian countries. In the process of active monitoring the complete autopsy of bird carcasses with description of organs and systems and sampling for laboratory diagnostics were performed. Droppings (pooled samples), parts of internal organs from dead and shot birds, blood (if possible) served as biological and pathological material for testing. While sampling, species were identified using an ornithological guide. The autopsy of dead waterfowl and birds shot for diagnostic purposes demonstrated a high worm burden of nematodes and cestodes. Two samples from European herring gulls were positive for avian influenza type A virus genome and subtype H13N6 was identified in one of them. Avian paramyxovirus serotype 1 (APMV-1), the agent of Newcastle disease, was found in one sample from gulls. The lakes of the Republic of Tyva are the most significant sites for sampling of biological material from wild birds, as the primary detection of highly pathogenic avian influenza virus in this territory is a serious signal of potential further virus spread and a precursor to a probable epizooty. Notwithstanding the absence of AIV very virulent isolate detections in wild bird populations the middle term prognosis for 2020 can be designated as cautious, as the avian influenza epidemic situation is deteriorating globally, especially in the European countries, and the threat of the virus introduction to the Russian territory with migratory birds still exists.

Under the conditions of intensive poultry farming and high concentration of poultry in a limited area mycoplasmoses cause significant economic damage to commercial poultry farming. Of greatest interest are respiratory mycoplasmosis and infectious synovitis, the causative agents of which are Mycoplasma gallisepticum and Mycoplasma synoviae, respectively. Considering that both diseases are included in the OIE list of notifiable diseases, it is necessary to perform constant control for their spread. The paper presents an analysis of serological test results for antibodies to mycoplasmosis agents in 2019. Six respiratory mycoplasmosis positive samples – and 129 infectious synovitis positive samples were detected when testing 2,401 chicken serum samples by the enzyme-linked immunosorbent assay (ELISA). The results of monitoring tests of samples received from 31 poultry farms (nine RF Subjects) indicate a decrease in the number of Mycoplasma gallisepticuminfected stocks and stable Mycoplasma synoviae situation. The respiratory mycoplasmosis epidemic situation on indoor poultry farms might have improved due to obtaining the poultry for commercial parent stocks from mycoplasmosis-free sources, better sanitary and hygienic conditions, elimination of the disease provoking factors, and the use of the disease-specific means of prevention in the parent broiler stocks. The infectious synovitis situation remains tense despite the fact that the number of Мycoplasma synoviae-infected farms decreased in comparison with the monitoring results for 2015–2018. Most seropositive stocks were detected on the layer farms (50.0% – in 2019). It seems appropriate to increase the amount of tests to be performed and the number of farms covered by testing in 2020 to more fully present the disease spread situation in commercial poultry farming in Russia.

FMD risk in the Russian Federation dictates the need for enhanced measures aiming to prevent the introduction of FMD virus and comprising systematic monitoring research and mass vaccination of susceptible animals in the buffer zone. Research into the development of vaccines for early protection confirm that their use induces the formation of virus-neutralizing antibodies in naturally susceptible animals in the outbreak area, which protects from FMD infection, limits its spread and contains it within the primary outbreak. Taking into account the high speed of the infection spread, such a control measure as using FMD vaccines which induce early protection should be adopted immediately after the occurrence of the outbreak. The article presents the results of the research into the formation of humoral immunity in naturally susceptible animals triggered by administration of inactivated emulsion FMD vaccines capable of ensuring early protection against type O FMD. Culture FMD virus of О/Primorsky/2012, О/Saudi Arabia/08 and О/Mongolia/2017 strains was used for vaccine production. Immunogenic activity of vaccines was tested in cattle, pigs, and sheep. It was found that monovalent emulsion FMD vaccine based on О/Mongolia/2017 strain induced the formation of virus-neutralizing antibodies in the quantity necessary to protect against the homologous strain in seven days after a single administration in the dose of 2 cm3 . Vaccines based on О/Saudi Arabia/08 and О/Primorsky/2012 FMDV strains can protect animals from infection with heterologous О/Mongolia/2017 strain at early stages if a double dose is administered. Vaccines based on the above-mentioned strains induce early immunity formation (seven days after vaccination) against type O FMD. We suggest using the given products in the zones of a higher risk of the virus introduction.

Mycoplasmosis control remains urgent in view of wide spread of bovine mycoplasmoses in the countries with intensive animal farming and trade relations between the Russian Federation and foreign partners including import of pedigree livestock and stud bull semen. Results of testing 1,186 biomaterial samples (blood, sera, nasal swabs, milk, preputial swabs, vaginal swabs, aborted and stillborn fetuses) collected from animals that demonstrated clinical signs of respiratory and reproductive disorders in 34 different regions of the Russian Federation for 2015–2018 are presented in the paper. The samples were tested with real-time polymerase chain reaction (rtPCR) for genomes of the following mycoplasmosis agents: Mycoplasma bovis, Mycoplasma bovigenitalium, Mycoplasma dispar. As a result, M. bovis genome was detected in 10.1% of the samples, M. bovigenitalium genome was detected in 8.6% of the samples and М. dispar genome was detected in 37.15% of the samples. Also, 927 semen samples submitted from Russian and foreign breeding farms were tested with PCR. Test results showed presence of M. bovis and M. bovigenitalium genomes in semen samples collected from native bull population. Presented data support Russian scientists’ conclusions on wide mycoplasmoses occurrence in cattle in the Russian Federation territory and risk of the disease agent introduction through semen import. All of these highlight the need for control of semen products as a source for mycoplasmosis spread as well as insufficiency of single testing of semen for granting the disease-free status to the breeding farm for genetic material marketing.

Porcine reproductive and respiratory syndrome (PRRS) being endemic and reported in the most countries in the world remains one of the most challenging diseases in pig industry. The main disease control measures include preventive vaccination and animal movement control within and outside the country as well as diagnostic testing of pigs in the population. Live and inactivated vaccines are used for specific prevention of porcine reproductive and respiratory syndrome. Complete and irreversible infectious agent inactivation with maximum epitope preservation and protective immunity in immunized animals are the main requirements for inactivated vaccines. Therefore, continuous improvement of methods for vaccine quality control at various vaccine production stages is of current importance. Results of development of the test system based on indirect liquid-phase enzyme-linked immunosorbent assay (ELISA) for PRRS virus antigen detection and activity testing in infectious and inactivated virus-containing cell cultures at intermediate stages of the vaccine production process are described in the paper. The test-system development process included purified and concentrated virus antigen as well as hyperimmune rabbit sera preparation. Specificity of purified and concentrated virus antigen was confirmed with real-time polymerase chain reaction. The developed test-system was shown to detect the virus antigen at initial infectivity titre of 4.87–7.21 lg TCID₅₀/cm³ corresponding to ELISA titre (dilution) of 1:4 up to 1:64. Methodical Guidelines for detection of porcine reproductive and respiratory syndrome virus antigen with indirect liquid-phase enzyme-linked immunosorbent assay (ELISA) (2019) were developed based on the work results, commissioned and approved by the FGBI “ARRIAH” Scientific Board.

Nidoviruses comprise the most complex grouping among other viruses with respect to their multiplicity, phylogeny, systematics, species identification, genetic relationships within a taxon, progressive variability. Like other single-stranded RNA viruses, nidoviruses have a relatively high ability to mutate and recombine, which allows them to quickly adapt to new hosts and new ecological niches. Although most of the known representatives of nidoviruses are associated with terrestrial hosts, more and more data has recently appeared on nidoviruses recovered from aquatic organisms. This review is the analysis of current data on the representatives of the order Nidovirales associated with aquatic animals. They are all included in the eight families based on the current classification of viruses. The most studied among them are members of the families Coronaviridae, Tobaniviridae and Roniviridae. Representatives of the other families of aquatic animal nidoviruses were identified using metagenomic deep sequencing (metagenomics), but their effect on the host organism has not yet been adequately studied. Data on the distribution of nidoviruses among aquatic animals in different global aquatic systems are presented, clinical signs of the disease are described, a brief description of nidoviruses and their genomes is given. Nidoviruses of aquatic animals as the earliest members of the animal kingdom are supposed to have played a possible role in the evolution of terrestrial animal nidoviruses. Therefore, aquatic animal nidoviruses could play a significant role in the formation of new natural reservoirs unknown to science, as well as in their interspecies transfer between marine, freshwater and terrestrial hosts.

Feline panleukopenia is a highly contagious viral disease of the members of the family Felidae caused by a DNA-virus of the family Parvoviridae. After infection, the virus is detected in saliva, nasal discharge, urine and feces; it is transmitted through excrements, water, food, and, according to some reports, by bloodsucking insects. The disease is characterized by gastrointestinal tract, central nervous system, hematopoietic tissue lesions. The paper describes the clinical signs of panleukopenia in cats in the territory of the Donetsk People’s Republic. Based on the records of a state-financed veterinary hospital and a charitable animal shelter located in the town of Yasinovataya, as well as those of private veterinary clinics and a veterinary office located in Donetsk, a classification of the clinical signs of the disease in pet and stray cats is suggested with the signs grouped according to their occurrence rate and by age groups. The paper provides evidence of feline panleukopenia occurrence in the territory of the Donetsk People’s Republic. In particular, the period from 2015 to 2018 was characterized by an increase in panleukopenia morbidity in stray animals; pet cats were also involved in the epidemic process. It was found that most of the clinical symptoms of panleukopenia were reported in cats of all age categories irrespective of their sex. Specific symptoms characteristic for different age groups were reported. It was found that panleukopenia morbidity rates for stray kittens aged 0–12 months were 1.8 times higher than those for pet kittens of the same age group. The disease was reported in adult pet cats 2.4 times less frequently than in stray cats. It is shown that the number of the disease cases in female cats is 1.5 times higher than in male ones. It is concluded that the violation of quarantine rules and insufficient vaccination coverage in cats are risk factors for the disease outbreak occurrence precipitating the multiple recurrent outbreaks of the disease.

Staphylococcus aureus (S. aureus) is a microorganism that causes a great number of diseases in humans and animals, including sepsis, pneumonia, food toxicoinfections, wound abscess, etc. Numerous studies on genotyping S. aureus strains isolated from humans, food and mastitis in cattle and small ruminants have been carried out. The lack of information on the genotyping of methicillin-susceptible S. aureus detected in monkeys served as a stimulus to conduct a similar research, since staphylococcal infections in the primates are widespread. The present study is devoted to molecular genetic testing of S. aureus isolated from different biological samples taken from monkeys and is based on typing of agr polymorphic locus which acts as a regulator of pathogenic gene expression. As a result of PCR analysis of 301 S. aureus isolates it was established that most of S. aureus belonged to agr IV (55%), and agr I (34%) was the second most group. Data resulting from the study differ from the results of other researchers published in literary sources, who performed typing of salmonella isolated from people with agr I prevailing. In conducting the study, neither distinct correlation between microbial isolation source and agr complex groups, nor relationship between the diseases and S. aureus group specificity were detected. Prevalence ratio of each agr group is nearly similar in S. aureus isolated from rhesus macaques and crab-eating macaques. But in hamadryas baboons and green monkeys II and III groups of agr complex were not detected.

Yeast-like fungi of the genus Candida are causative agents of the infectious pathology of the mucous membrane of the gastrointestinal, respiratory, urogenital tracts and skin of mammals, sepsis, and disseminated infection in birds. The search and testing of multilevel algorithms for biofilm identification when exposed to chemotherapeutic and disinfectant drugs for blocking the synthesis or destruction of the intercellular matrix in the development of superficial, deep and systemic candidiasis of animals are relevant for developing and improving diagnostic and antiepidemic measures. It was established that the formation of biofilm heterogeneous structure comprises multiple stages implementing the processes of intercellular communication due to the synthesis of a polymer matrix composites. Optical microscopy revealed a three-dimensional structure of biofilms in the form of a dense network consisting of yeast cells, hyphal and pseudohyphalic forms surrounded by an intercellular polymer matrix. Candida spp. pathogenicity factors contribute to infection of susceptible species due to adhesion, invasion, secretion of hydrolases, dimorphism. Formation of mono-species or poly-species biofilms of microorganisms, including Candida spp., causes the development of superficial, deep and systemic candidiasis. Detection of a large amount of yeast and micellar phases in C. albicans and C. africana isolates was a differential sign of a significant degree of colonization of the mucous membranes of the larynx, pharynx, and tonsils in case of local and systemic pathologies in pigs. The results of studies of the biofilm heterogeneous structure and phenotypic signs of yeast-like fungi can be used in a comparative study of biological characteristics and the identification of common patterns and differential signs of microorganisms, optimization of mycological diagnostics, and also in the development of antimycotic drugs.

Results of mycotoxicological survey of representative samples of feed and procured wheat, barley, oat and maize grain batches are demonstrated. The samples were submitted by the Veterinary Service officials, livestock farmers and feed mill operators, agricultural producers, specialized commercial business operators and farm owners in seven Federal Districts of the Russian Federation in 2009-2019. Similar amounts of wheat and barley grain samples were received from the Central, Volga, Ural and Siberian Federal Districts. The amount of wheat samples delivered from the Southern Federal District prevailed over the number of barley samples, and the maize samples were mostly delivered from the regions of the Central Federal District. Fusarium toxins including T-2 toxin, diacetoxyscirpenol, deoxynivalenol, zearalenone and fumonisins of group B as well as alternariol, ochratoxin A, citrinin, aflatoxin В1, sterigmatocystin, cyclopiazonic acid, mycophenolic acid, ergot alkaloids and emodin were detected and measured according to the validated competitive ELISA procedure. Generalization of the results demonstrated domination of fusarium toxins and active involvement of alternariol in the contamination of all types of feed grains as well as high occurrence of emodin in ear cereals and increased occurrence of T-2 toxin and ochratoxin A in barley. Shift of medians and 90%-percentile of the basic contaminants to lower values as compared to mean and maximal ones was reported thus being indicative of their possible accumulation at the levels outside the typical range. The highest levels of T-2 toxin, deoxynivalenol and ochratoxin A as well as 90%-percentile values exceeded the acceptable levels. The maize grains demonstrated the whole complex of the tested fusarium toxins with the prevalence of T-2 toxin, deoxynivalenol, zearalenone and fumonisins; and the maximal amounts of these mycotoxins by several times exceeded the accepted regulatory levels. Diacetoxyscirpenol, aflatoxin В1, sterigmatocystin, cyclopiazonic acid and ergot alkaloids are classified as rare feed grain contaminants. High prevalence of alternariol and emodin known as “diarrhea factor” as well as maize grain contamination with mycophenolic acid (mycotoxin having an immunosuppressive effect) are for the first time reported in this paper. These data support the need of their introduction in the group of regulated substances significant for public health. Original monitoring data systematized and summarized in the paper are given in electronic format in section Additional materials.

Part 1: Veterinary Science Today. 2020; 1 (32): 60–65. DOI: 10.29326/2304-196X-2020-1-32-60-65.

Peer-review of monograph of V. V. Makarov, N. Ya. Makhamat, A. M. Gulyukin, M. I. Gulyukin “Anthrax: Modern Knowledge and Global Occurrence”. М.: Agentstvo tvorcheskikh tekhnologiy, 2019. 145 p.: il. ISBN 978-5-6042239-6-3.

In memory of Alexander N. Burdov.