Seroprevalence of Toxoplasma gondii in wild boars (Sus scrofa) hunted in Ukraine.

Toxoplasma gondii is an important zoonotic parasite worldwide, but it has received limited attention in Ukraine. A seroepidemiological study was conducted and samples from 452 wild boars that had been hunted in 2006-2011 in 23 of the 25 regions of Ukraine were tested to estimate T. gondii seroprevalence. A locally available commercial enzyme-linked immunosorbent assay (ELISA) was used for the investigation. Additionally, we tested 92 of the sera using a widely used commercial multi-species ELISA and an indirect immunofluorescence antibody test (IFAT). With the locally available ELISA, 35 of the 452 wild boars tested positive, yielding a seroprevalence estimate of 7.7% (95% confidence interval 5.5-10.5). The seropositive wild boars originated from eight of the regions. Using the majority criteria, 10/92 samples tested using both ELISAs and the IFAT were considered positive, yielding an estimated seroprevalence of 10.9% within the subset of samples. The highest seroprevalence was observed in wild boars hunted in Luhans'k (30.0%), Odesa (17.7%) and Kharkiv (12.7%). Seroprevalence was higher in older animals (13.3% for age group >12 months and 7.7% for age group ≤12 months). This is the first seroepidemiological study of T. gondii in wild boars in Ukraine. Assuming that seropositivity indicates presence of infectious parasites in the tissues, eating undercooked meat of wild boars hunted in Ukraine could be a potential source of infection to other hosts, including humans.

Genetic Diversity of Porcine Circovirus 2 in Wild Boar and Domestic Pigs in Ukraine.

Porcine circovirus type 2 (PCV2) is responsible for a number of porcine circovirus-associated diseases (PCVAD) that can severely impact domestic pig herds. For a non-enveloped virus with a small genome (1.7 kb ssDNA), PCV2 is remarkably diverse, with eight genotypes (a-h). New genotypes of PCV2 can spread through the migration of wild boar, which are thought to infect domestic pigs and spread further through the domestic pig trade. Despite a large swine population, the diversity of PCV2 genotypes in Ukraine has been under-sampled, with few PCV2 genome sequences reported in the past decade. To gain a deeper understanding of PCV2 genotype diversity in Ukraine, samples of blood serum were collected from wild boars (n = 107) that were hunted in Ukraine during the November-December 2012 hunting season. We found 34/107 (31.8%) prevalence of PCV2 by diagnostic PCR. For domestic pigs, liver samples (n = 16) were collected from a commercial market near Kharkiv in 2019, of which 6 out of 16 (37%) samples were positive for PCV2. We sequenced the genotyping locus ORF2, a gene encoding the PCV2 viral capsid (Cap), for 11 wild boar and six domestic pig samples in Ukraine using an Oxford Nanopore MinION device. Of 17 samples with resolved genotypes, the PCV2 genotype b was the most common in wild boar samples (10 out of 11, 91%), while the domestic pigs were infected with genotypes b and d. We also detected genotype b/d and b/a co-infections in wild boars and domestic pigs, respectively, and for the first time in Ukraine we detected genotype f in a wild boar from Poltava. Building a maximum-likelihood phylogeny, we identified a sublineage of PCV2 genotype b infections in both wild and domestic swine, suggesting a possible epizootic cluster and an ecological interaction between wild boar and domestic pig populations in northeastern Ukraine.

Seroprevalence of Enzootic Teschen Disease in the Wild Boar Population in Ukraine.

Teschen disease is an acute fatal enterovirus encephalomyelitis of pigs, characterized by a range of central nervous system disorders. The cause of porcine enterovirus encephalomyelitis is the picornavirus porcine teschovirus-1 (PTV-1). There are at least 12 disctinct serotypes of PTVs, where PTV-2 to PTV-12 serogroups are associated with other forms of disease (Talfan disease or poliomyelitis suum) or benign enzootic paresis. Combined, PTVs have been found to have a high seroprevalence, up to 65%, in healthy pig populations in Europe. PTVs have also been detected in wild boar, including the divergent PTV-13 serogroup; wild suids may represent a sylvatic reservoir capable of carrying the virus long distances. In Ukraine, Teschen disease is widespread and causes lethal disease in domestic pigs. To understand temporal and geographical distribution of Teschen disease virus (PTV-1) in wild boar in Ukraine (2001-2013), we analyzed seroprevalence of 6840 blood serum samples from hunted suids using a virus microneutralization assay. A total of 1364 samples (19.9%) were seropositive, with average antibody titer ratios 5.89 ± 0.03 log2 (range 5-12 log2). Teschen seroprevalence was temporally and geographically concentrated in the northern and western regions of Ukraine, corresponding to forested regions (polissya) and overlapping with wild boar populations and habitats, suggesting endemicity in wild boar. The virus sporadically emerged in central, southern, and eastern forested regions, suggesting long-distance movement of infected wild suids. Thus, wild boar should be monitored for potential transboundary spread in forested and mountain regions and spillover of PTVs to domestic swine populations.

Supporting control programs on African swine fever in Ukraine through a knowledge, attitudes, and practices survey targeting backyard farmers.

A Knowledge, Attitudes, and Practices (KAP) questionnaire was designed to collect information on farmers' knowledge of ASF and their practices surrounding that could impact the spread of the disease. The questionnaire was distributed, and data collected, from 233 backyard farmers from five selected Oblasts (Rivne, Kharkiv, Odessa, Zakarpattia and Kiev). Kruskal-Wallis tests were conducted to identify factors that could influence knowledge, and Dunn tests were performed to determine differences between groups when the Kruskal-Wallis tests were significant. Spearman tests were carried out to explore the association between knowledge and risky practices. Results show that comprehensive knowledge on ASF is not common in backyard farmers and that risky practices that influence the spread of ASF are regularly performed. Of the respondents, 47% felt well-informed about how ASF can be transmitted and 31.8% felt confident about recognizing clinical signs of ASF. The independent variable "Oblast" was identified as a significant factor (p = 0.0015) associated with differences in knowledge on clinical signs. We demonstrated statistically significant differences of knowledge between backyard farmers from different Oblasts. Knowledge of preventive measures was positively correlated with risky handling practices related to edible pork products (p = 0.0053) and non-edible pork products (p = 0.0417). In conclusion, our results show that backyard farmers have knowledge gaps on ASF and practice various risky behaviours that might favour the spread of the disease in Ukraine. There are regional differences in ASF knowledge and risky practices that should be taken into consideration in future evidence-based ASF prevention and control programs, including public awareness activities.

Presence of Antibodies Against Leptospira interrogans Serovar hardjo in Serum Samples from Cattle in Ukraine.

The article presents data on serological studies of 573 sera samples of cattle that were collected from the farms affected by leptospirosis in different regions of Ukraine in the period of 2014-2015. Samples were investigated by the microscopic agglutination test (MAT), which was conducted within eight serological groups of Leptospira and nine serovars: Sejroe (serovars polonica and hardjo), Hebdomadis (serovar kabura), Tarassovi (serovar tarassovi), Pomona (serovar pomona), Grippotyphosa (serovar grippotyphosa), Canicola (serovar canicola), Icterohaemorrhagiae (serovar copenhageni), and Australis (serovar bratislava). The circulation of L. interrogans serovar hardjo among cattle has been observed in all 11 regions of Ukraine investigated within 25.8-60.0% of the leptospirosis-positive serum samples in these regions. Antibodies in the cattle sera against serovar hardjo (serogroup Sejroe) were detected in 139 of the 370 cows reacting positively in MAT. Overall, they were detected in 24.3% animals out of the total of 573 cows investigated. These are the preliminary results, however, in our opinion, they should allow to include the serovar hardjo in a standard panel of strains for MAT in Ukraine.The article presents data on serological studies of 573 sera samples of cattle that were collected from the farms affected by leptospirosis in different regions of Ukraine in the period of 2014­2015. Samples were investigated by the microscopic agglutination test (MAT), which was conducted within eight serological groups of Leptospira and nine serovars: Sejroe (serovars polonica and hardjo), Hebdomadis (serovar kabura), Tarassovi (serovar tarassovi), Pomona (serovar pomona), Grippotyphosa (serovar grippotyphosa), Canicola (serovar canicola), Icterohaemorrhagiae (serovar copenhageni), and Australis (serovar bratislava). The circulation of L. interrogans serovar hardjo among cattle has been observed in all 11 regions of Ukraine investigated within 25.8­60.0% of the leptospirosis-positive serum samples in these regions. Antibodies in the cattle sera against serovar hardjo (serogroup Sejroe) were detected in 139 of the 370 cows reacting positively in MAT. Overall, they were detected in 24.3% animals out of the total of 573 cows investigated. These are the preliminary results, however, in our opinion, they should allow to include the serovar hardjo in a standard panel of strains for MAT in Ukraine.

Implementation of a Regional Training Program on African Swine Fever As Part of the Cooperative Biological Engagement Program across the Caucasus Region.

A training and outreach program to increase public awareness of African swine fever (ASF) was implemented by Defense Threat Reduction Agency and the Ministries of Agriculture in Armenia, Georgia, Kazakhstan, and Ukraine. The implementing agency was the company SAFOSO (Switzerland). Integration of this regional effort was administered by subject matter experts for each country. The main teaching effort of this project was to develop a comprehensive regional public outreach campaign through a network of expertise and knowledge for the control and prevention of ASF in four neighboring countries that experience similar issues with this disease. Gaps in disease knowledge, legislation, and outbreak preparedness in each country were all addressed. Because ASF is a pathogen with bioterrorism potential and of great veterinary health importance that is responsible for major economic instability, the project team developed public outreach programs to train veterinarians in the partner countries to accurately and rapidly identify ASF activity and report it to international veterinary health agencies. The project implementers facilitated four regional meetings to develop this outreach program, which was later disseminated in each partner country. Partner country participants were trained as trainers to implement the outreach program in their respective countries. In this paper, we describe the development, execution, and evaluation of the ASF training and outreach program that reached more than 13,000 veterinarians, farmers, and hunters in the partner countries. Additionally, more than 120,000 booklets, flyers, leaflets, guidelines, and posters were distributed during the outreach campaign. Pre- and post-ASF knowledge exams were developed. The overall success of the project was demonstrated in that the principles of developing and conducting a public outreach program were established, and these foundational teachings can be applied within a single country or expanded regionally to disseminate disease information across borders; overall, this method can be modified to raise awareness about many other diseases.