Aujeszky's Disease Virus Research Articles

PSVIII-22 Genome-wide association studies for immune response and resilience to Aleutian disease in mink

Abstract Aleutian disease (AD) is a severe health issue for the mink industry, causing increased mortality and adversely impacting several economically important traits, resulting in substantial economic losses. AD has been characterized as an immune complex disease, which indicates that the greater concentrations of anti-AMDV antibodies produced more harmful to the host. Resilience is defined as the ability the animals have to maintain their performance under exposure to disease-causing agents and disruptions. The phenotypic selection of AD-resilient mink based on immune response and/or indicator traits is practiced by some mink farms, but the genetic architecture of immune response and resilience to AD has not been widely explored. In this study, mink (n = 1,411) from the Canadian Centre for Fur Animal Research (Truro, Nova Scotia, Canada), which is an AD-positive facility, were used to detect potential genomic regions and genes related to immune response and feed-intake-related resilience of mink to AD through genome-wide association studies (GWAS) analyses. The studied individuals were genotyped using the Axiom Affymetrix Mink 70K single nucleotide polymorphism (SNP) panel. The studied phenotypes included two immune response traits measured by antigen-based enzyme-linked immunosorbent assay (ELISA-G) and iodine agglutination test (IAT) and two feed-intake-related resilience traits measured by the daily variation in feed intake (Varf) and proportion of off-feed d (DOF). The de-regressed breeding values were derived from the estimated breeding values for each trait and utilized as pseudo-phenotypes to perform GWAS analyses using a single SNP univariate mixed linear animal model in SNP1101 software. A total of 17 significant [false-discovery-rate-adjusted-p-value (q) < 0.01] SNPs were detected to be associated with ELISA-G, and 141 genes were annotated from the detected SNPs. Among all the annotated genes, three genes, MPIG6B, RUNX2, and C4A, were found to have important roles in immune-mediated responses to AD. Eight SNPs were detected to be significantly associated with IAT, and 44 genes were annotated from these eight SNPs. Two (TNFRSF11A and C4A) of the 44 genes were involved in the immune system process. For DOF, seven significant associated SNPs were detected, and 42 genes were annotated from these SNPs. Two annotated genes, ADCY7 and CNDP2, were related to feed intake or appetite. Five significant (q < 0.05) overrepresented gene ontology enrichment terms, including TAP complex, Classical-complement-pathway C3/C5 convertase complex, MHC class I peptide loading complex, extracellular region, and ABC-type peptide transporter activity, were detected for ELISA-G, and they have important roles in the adaptive immune response or complement system. The newly detected significant SNPs and identified candidate genes in this study would provide a better understanding of the genetic architecture and biological mechanisms underlying AD resilience in mink, which offers an opportunity for increasing the resilience of mink to AD using marker-assisted/genomic selection in mink.

200 Transcriptomics analyses of mink spleen infected by Aleutian Disease

Abstract Aleutian disease (AD) is one of the most challenging mink diseases that causes high mortality and affects several economically important traits. Aleutian mink disease virus (AMDV) targets multiple organs and among them, the spleen is one of the major targeted organs. Transcriptomics has been widely used to reveal genes and biological pathways and identify biomarkers for early detection or prevention of diseases. This study aimed to identify genes and pathways related to the host response to AD infection in the spleen via transcriptomic study. Twelve (3 sets of 4 full-sibs) AMDV-free black male mink were housed in the clean section of the animal housing facility at the AD Research Centre (Truro, NS, Canada) between 4 to 7 d before inoculation. Before inoculation, sampling, and euthanasia, animals were anesthetized. The viral inoculum was a 10% (W/V) passage 2 of AMDV prepared from the spleens of mink infected with a local strain and stored at − 80 °C. Animals were intranasally inoculated under sedation with 60 µL of the viral homogenate, corresponding to approximately 300 to 700 ID50. Infected mink were humanely euthanized and the spleen tissues were collected for RNA isolation at 24h (d 1), 48h (d 2), and d 7. Libraries were prepared from total RNA using the Illumina TruSeqTM RNA kit, and were sequenced using the HiScanSQ platform, producing 101 bp reads from each end. The raw RNA-Seq data were cleaned using Cutadapt V1.4.2 and after the cleaning process was completed, 942,803,540 paired-end reads remained to be used in downstream analysis. Differential gene expression analyses revealed most (168) significant differentially expressed (DE) genes between d 1 and d 0 (Control or ADMV-free mink) while fewer DE genes (23) were between d 7 and d 0 (Table 1; Figure 1). A total of 19 DE genes are identified between three pair comparisons and seven of them are directly involved in immune response (FGL2, TLF8, LRP1, SERPINB9, MSR1, C3, PLA2R1, and XCR1). Gene enrichment analyses revealed pathways related to innate immune and fat metabolism which are important for the host mechanism to fight against infections. One of the important pathways is neutrophil degranulation which functions as aiding in the elimination of pathogens and the initiation of the inflammatory process. In conclusion, the current study provides insight into the transcriptomic profiles of the spleen in mink infected with AD. Identified candidate genes might be used for functional studies or as prior information for markers or genomic selection against AD in mink. Further studies in other tissues or single-cell RNA sequencing might deliver more comprehensive pictures of host responses to AD infections.

Prevalence of Aleutian Mink Disease Virus (AMDV) in Free-Ranging American Mink from Biebrza and Narew National Parks (Poland)-An Epidemiological Concern.

Aleutian Mink Disease Virus (AMDV) is the causative agent of Aleutian disease (AD). This progressive and chronic disorder significantly impacts the mink breeding industry, affecting farmed and free-ranging American and European mink. This study investigated AMDV variants isolated from free-ranging American mink in northeastern Poland. Between 2018 and 2019, 26 spleen samples were collected from mink in Narew National Park (NNP) and Biebrza National Park (BNP). DNA was extracted and subjected to PCR to amplify the NS1 gene, followed by sequencing and phylogenetic analysis. The NS1 gene was detected in 50% of samples from NNP minks and in 30% of samples from BNP minks, with an overall prevalence of 42.31%; these findings align with global data and indicate serious ecological and health concerns. Ten closely related AMDV variants and one distinct variant were identified. The grouped variants exhibited high genetic homogeneity, closely related to strains found in mink from the USA, Germany, Greece, Latvia, and Poland; meanwhile, the distinct variant showed similarities to strains found in mink from Finland, Denmark, China, Poland, and Latvia, suggesting multiple infection sources. These findings, consistent with data from Polish mink farms, indicate significant genetic similarity between farmed and wild mink strains, suggesting potential bidirectional transmission. This underscores the importance of a One Health approach, emphasizing the interconnectedness of human, animal, and environmental health. Continuous surveillance and genetic studies are crucial for understanding AMDV dynamics and mitigating their impacts. Measures to reduce transmission between farmed and wild mink populations are vital for maintaining mink health and ecosystem stability.

Microbiological and decomposition analysis of mass mink burial sites during the COVID-19 pandemic

In 2020, Denmark buried approximately four million culled, farmed mink in mass graves treated with slaked lime due to widespread SARS-CoV-2 infections. After six months, environmental concerns prompted the exhumation of these cadavers. Our analysis encompassed visual inspections, soil pH measurements, and gas emission assessments of the grave environment. Additionally, we evaluated carcasses for decay status, cadaverine content, and the presence of various pathogens, including SARS-CoV-2 and mink coronavirus. Our findings revealed minimal microbial activity and limited carcass decomposition. Although viral RNA from SARS-CoV-2 and mink coronavirus, along with DNA from Aleutian mink disease virus, were detected, the absence of infectious SARS-CoV-2 in cell culture assays suggests slow natural degradation processes. This study provides critical insights for future considerations in managing mass burial scenarios during outbreaks of livestock-associated zoonotic pathogens.

Isolation, structural determination, and antiviral activities of a novel alanine-conjugated polyketide from Talaromyces sp.

Antiviral agents are highly sought after. In this study, a novel alkylated decalin-type polyketide, alaspelunin, was isolated from the culture broth of the fungus Talaromyces speluncarum FMR 16671, and its structure was determined using spectroscopic analyses (1D/2D NMR and MS). The compound was condensed with alanine, and its absolute configuration was determined using Marfey's method. Furthermore, the antiviral activity of alaspelunin against various viruses was evaluated, and it was found to be effective against both severe acute respiratory syndrome coronavirus 2 and pseudorabies (Aujeszky's disease) virus, a pathogen affecting pigs. Our results suggest that this compound is a potential broad-spectrum antiviral agent.

The Reproduction Number of Swine Viral Respiratory Diseases: A Systematic Review.

Diseases in the swine industry can cause significant economic and health impacts. This review examines R0 estimates for respiratory diseases in pigs, assessing variations and comparing transmission risks within and between farms. A literature search of three databases aggregated peer-reviewed research articles on swine viral respiratory diseases' R0 values. The study focused on seven diseases: Aujeszky's disease (AD), Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), Circovirus, Influenza A (IA), Encephalomyocarditis Virus (EV), Classical Swine Fever (CSF), and African Swine Fever (ASF). R0 values were estimated for transmission within and between herds/farms using various methods, from complex mathematical models to simple calculations. Data primarily came from disease surveillance and laboratory experiments. The median R0 for within-herd and between-herd transmission was 10 and 3.31 for AD, 2.78 and 1.14 for PRRSV, 5.9 and 0.89 for Circovirus, 1.75 and 1.6 for CSF, and 3.94 and 3.15 for ASF. For IA and EV, only within-herd R0 values were estimated at 8.65 and 1.3, respectively. Diseases with high R0 values highlight the need for prompt detection and response to outbreaks. Continuous monitoring and evaluation of pathogen transmissibility are crucial for enhancing disease surveillance and reducing the impact of livestock diseases.

A qualitative risk assessment of imports of animal feed as a potential pathway for Aujeszky's disease virus incursion

Aujeszky's disease (AD) is a highly contagious disease of pigs that primarily transmits by respiratory and oral routes. Evidence from recent outbreaks suggests that some swine viruses can survive in contaminated animal feed, thus posing a risk of entry via imports from other countries. To this end, a qualitative risk assessment was undertaken to determine the risk of introduction of AD virus (ADV) and infection of pigs via this route to determine if contaminated animal feed is a viable pathway for the spread of ADV. The feed categories investigated were soya bean/meal/oilcake, pet food, choline/lysine and spray dried porcine plasma. These were chosen based on their use in animal feed and the available data on viral contamination. The overall probability of an animal becoming infected from the importation of feed contaminated with ADV was estimated as Negligible or Very Low for all feed categories. The uncertainty associated with the estimates was assessed as Medium, due to the lack of data around the mechanisms that ADV could contaminate feedstuffs and for infection of susceptible animals from ADV infected feed.

Diverse amdoparvoviruses infection of farmed Asian badgers (Meles meles).

Amdoparvoviruses infect various carnivores, including mustelids, canids, skunks, and felids. Aleutian mink disease virus (AMDV) belongs to the prototypical species Amdoparvovirus carnivoran1. Here, we identified a novel amdoparvovirus in farmed Asian badgers (Meles meles), and we named this virus "Meles meles amdoparvovirus" (MMADV). A total of 146 clinical samples were collected from 134 individual badgers, and 30.6% (41/134) of the sampled badgers tested positive for amdoparvovirus by PCR. Viral DNA was detected in feces, blood, spleen, liver, lung, and adipose tissue from these animals. Viral sequences from eight samples were determined, five of which represented nearly full-length genome sequences (4,237-4,265 nt). Six serum samples tested positive by PCR, CIEP, and IAT, four of which had high antibody titers (> 512) against AMDV-G. Twenty-six of the 41 amdoparvovirus-positive badgers showed signs of illness, and necropsy revealed lesions in their organs. Sequence comparisons and phylogenetic analysis of the viral NS1 and VP2 genes of these badger amdoparvoviruses showed that their NS1 proteins shared 62.6%-88.8% sequence identity with known amdoparvoviruses, and they clustered phylogenetically into two related clades. The VP2 proteins shared 76.6%-97.2% identity and clustered into two clades, one of which included raccoon dog and arctic fox amdoparvovirus (RFAV), and the other of which did not include other known amdoparvoviruses. According to the NS1-protein-based criterion for parvovirus species demarcation, the MMADV isolate from farm YS should be classified as a member of a new species of the genus Amdoparvovirus. In summary, we have discovered a novel MMADV and other badger amdoparvoviruses that naturally infect Asian badgers and are possibly pathogenic in badgers.

Molecular Detection of Porcine Cytomegalovirus, Porcine Parvovirus, Aujeszky Disease Virus and Porcine Reproductive and Respiratory Syndrome Virus in Wild Boars Hunted in Serbia during 2023.

Porcine cytomegalovirus (PCMV) infection is widespread worldwide and has a high prevalence in swine herds, especially in countries with intensive swine production. PCMV is zoonotic and can impact xenotransplants. It is the third swine virus known to be zoonotic, following swine influenza virus (influenza A) and hepatitis E virus genotype 3 (HEVgt3 or HEV-3). Wild boars, serving as reservoirs for various pathogens, including PCMV, pose a risk to both the pig industry and public health. This study aimed to investigate PCMV infection in Serbian wild boars using real-time PCR and assess other viral infections. We also tested samples for the presence of other viral infections: Aujeszky disease virus (ADV), Porcine parvovirus (PPV) and Porcine reproductive respiratory syndrome (PRRSV). Samples from 50 wild boars across 3 districts were tested. Results showed 8% positivity for PCMV DNA, with females showing higher infection rates. Porcine parvovirus (PPV) was detected in 56% of samples, while Porcine reproductive respiratory syndrome virus (PRRSV) was absent. ADV was found in 18% of samples, primarily in younger animals. This research contributes to understanding PCMV prevalence in Serbian wild boars and emphasizes the importance of monitoring viral infections in wild populations, considering the potential zoonotic and economic implications.

Development and characterization of a novel specific monoclonal antibody against mink enteritis virus and its antigen epitope analysis

This study prepared a novel monoclonal antibody (MAb) against mink enteritis parvovirus (MEV) and identified its antigen epitope. The antibody subclass is identified as IgG1, the titers of the MAb is up to 1:1 × 106 and keeps stably after low-temperature storage for 9 months or 11 passages of the MAb cells. The MAb can specifically recognize MEV in the cells in IFA, but not Aleutian disease virus (ADV) or canine distemper virus (CDV). Its antigen epitope was identified as a polypeptide containing 5 key amino acids (378YAFGR382) and the homology in 20 MEV strains, 4 canine parvovirus strains, and 4 feline panleukopenia virus strains was 100%. This study supplies a biological material for developing new methods to detect MEV.

Establishment and application of multiplex PCR for rapid detection of three mink diarrhea-associated viruses

In this report, a multiplex PCR method was developed for the detection of three diarrhea-associated viruses in mink, including circovirus (MCV), bocavirus (MBoV), and enteritis virus (MEV). Three compatible sets of primers specific for each virus were designed respectively based on their conserved sequences. After optimization of the crucial factors such as primer concentration and annealing temperature in single and multiple amplification, three specific fragments were simultaneously amplified with the highest sensitivity and specificity in one PCR reaction. The fragments amplified were 259 bp (MCV)，455 bp (MBoV) and 671 bp (MEV). The sensibility of this one-step multiplex PCR is about 10 times lower than that of regular singleplex PCR. There were no cross-reactions with some relevant pathogens like mink coronavirus, canine distemper virus, and aleutian mink disease virus. In our study we analyzed viral DNA in mink fecal samples by multiplex PCR assay from China, which revealed the occurrence of MCV, MBoV, and MEV as 3.1 %, 5.7 %, and 9.8 %, respectively. The testing results of multiplex PCR agreed with the singleplex PCR results with a coincidence rate of 100 %. These results indicated that the method could provide technical support for rapid detection of the three diarrhea-associated viruses, and epidemiological investigation of mink viral diarrhea.

Strong selection signatures for Aleutian disease tolerance acting on novel candidate genes linked to immune and cellular responses in American mink (Neogale vison)

Aleutian disease (AD) is a multi-systemic infectious disease in American mink (Neogale vison) caused by Aleutian mink disease virus (AMDV). This study aimed to identify candidate regions and genes underlying selection for response against AMDV using whole-genome sequence (WGS) data. Three case–control selection signatures studies were conducted between animals (N = 85) producing high versus low antibody levels against AMDV, grouped by counter immunoelectrophoresis (CIEP) test and two enzyme-linked immunosorbent assays (ELISA). Within each study, selection signals were detected using fixation index (FST) and nucleotide diversity (θπ ratios), and validated by cross-population extended haplotype homozygosity (XP-EHH) test. Within- and between-studies overlapping results were then evaluated. Within-studies overlapping results indicated novel candidate genes related to immune and cellular responses (e.g., TAP2, RAB32), respiratory system function (e.g., SPEF2, R3HCC1L), and reproduction system function (e.g., HSF2, CFAP206) in other species. Between-studies overlapping results identified three large segments under strong selection pressure, including two on chromosome 1 (chr1:88,770–98,281 kb and chr1:114,133–120,473) and one on chromosome 6 (chr6:37,953–44,279 kb). Within regions with strong signals, we found novel candidate genes involved in immune and cellular responses (e.g., homologous MHC class II genes, ITPR3, VPS52) in other species. Our study brings new insights into candidate regions and genes controlling AD response.

Genome-wide association studies for economically important traits in mink using copy number variation

Copy number variations (CNVs) are structural variants consisting of duplications and deletions of DNA segments, which are known to play important roles in the genetics of complex traits in livestock species. However, CNV-based genome-wide association studies (GWAS) have remained unexplored in American mink. Therefore, the purpose of the current study was to investigate the association between CNVs and complex traits in American mink. A CNV-based GWAS was performed with the ParseCNV2 software program using deregressed estimated breeding values of 27 traits as pseudophenotypes, categorized into traits of growth and feed efficiency, reproduction, pelt quality, and Aleutian disease tests. The study identified a total of 10,137 CNVs (6968 duplications and 3169 deletions) using the Affymetrix Mink 70K single nucleotide polymorphism (SNP) array in 2986 American mink. The association analyses identified 250 CNV regions (CNVRs) associated with at least one of the studied traits. These CNVRs overlapped with a total of 320 potential candidate genes, and among them, several genes have been known to be related to the traits such as ARID1B, APPL1, TOX, and GPC5 (growth and feed efficiency traits); GRM1, RNASE10, WNT3, WNT3A, and WNT9B (reproduction traits); MYO10, and LIMS1 (pelt quality traits); and IFNGR2, APEX1, UBE3A, and STX11 (Aleutian disease tests). Overall, the results of the study provide potential candidate genes that may regulate economically important traits and therefore may be used as genetic markers in mink genomic breeding programs.

Identifying selection signatures for immune response and resilience to Aleutian disease in mink using genotype data.

Aleutian disease (AD) brings tremendous financial losses to the mink industry. Selecting AD-resilient mink has been conducted to control AD. Such selections could have altered the patterns of genetic variation responding to selection pressures. This study aimed to identify selection signatures for immune response (IRE) and resilience to AD. A total of 1,411 mink from an AD-positive facility were used. For IRE, 264 animals were categorized according to the combined results of enzyme-linked immunosorbent assay (ELISA) and counterimmunoelectrophoresis (CIEP). For resilience, two grouping methods were used: 1) general resilience performance (GRP, n = 30) was evaluated based on the feed conversion ratio, Kleiber ratio, and pelt quality; and 2) female reproductive performance (FRP, n = 36) was measured based on the number of kits alive 24h after birth. Detection methods were the pairwise fixation index, nucleotide diversity, and cross-population extended haplotype homozygosity. A total of 619, 569, and 526 SNPs were identified as candidates for IRE, GRP, and FRP, respectively. The annotated genes were involved in immune system process, growth, reproduction, and pigmentation. Two olfactory-related Gene Ontology (GO) terms were significant (q < 0.05) for all traits, suggesting the impact of AD on the sense of smell of infected mink. Differences in detected genes and GO terms among different color types for IRE indicated variations in immune response to AD among color types. The mitogen-activated protein kinase (MAPK) signaling pathway was significant (q < 0.05) for FRP, suggesting that AD may disrupt MAPK signaling and affect FRP. The findings of this research contribute to our knowledge of the genomic architecture and biological mechanisms underlying AD resilience in mink.

Aleutian disease: Risk factors and ImmunAD strategy for genetic improvement of tolerance in American mink (Neogale vison).

Aleutian disease (AD) is a devastating infectious disease in American mink (Neogale vison) industry caused by Aleutian mink disease virus (AMDV). Two crucial steps toward controlling infectious diseases in farm animals are: (i) assessment of the infection risk factors to minimize the likelihood of infection and (ii) selection of animals with superior immune responses against pathogens to build tolerant farms. This study aimed to investigate AD risk factors and evaluate a novel "ImmunAD" approach for genetic improvement of AD tolerance. Phenotypic records and pedigree information of 1,366 and 24,633 animals were included in this study. The risk of animal's age, sex, color type, and year of sampling on AMDV infection was assessed using a logistic regression model and counter immune-electrophoresis (CIEP) test results. ImmunAD phenotype was calculated based on AMDVG enzyme-linked immunosorbent assay (ELISA) and CIEP test results, and breeding values for ImmunAD were estimated using an animal model. Animals were classified into high-coordinated (HCIR), average-coordinated (ACIR), and low-coordinated immune responders (LCIR) using ImmunAD's breeding values, and the impact of selection of HCIR on live grade of pelt quality (PQ), harvest weight (HW), and harvest length (HL) breeding values were evaluated. Age of > 1 year, male sex, and year of sampling were identified as significant risk factors of AD (p < 0.05). A moderate-to-high heritability (0.55±0.07) was estimated for ImmunAD, while a higher heritability was observed among the CIEP-positive animals (0.76±0.06). Significantly higher breeding values were observed for PQ and HL among HCIR than those for LCIR and ACIR (p < 0.05). Our findings indicate the critical role of male breeders in AD distribution within mink farms. Regular screening of AD in male breeders before pairing them with females during breeding seasons can help disease control. ImmunAD strategy can be applied to genetic improvement of AD tolerance, with favorable impacts on some growth and production traits. Higher genetic gains can be achieved in populations with higher AD seroprevalences.

PSXII-9 Identifying Selection Signatures for Immune Response and Resilience to Aleutian Disease in Mink Using Genotypes Data

Abstract Aleutian disease (AD), a severe immune-complex disease, brings tremendous financial losses to the mink industry. Resilience is the ability of an animal to minimize the influences caused by disturbances and maintain its performance under pathogen exposure. Many AD-positive farms have selected AD-resilient mink based on AD tests and/or AD-resilience indicator traits, such as growth, pelt quality, and reproduction. Selection based on these indicator traits may have changed the patterns of genetic variation and potentially represent a genomic signature that can be used to identify genes subjected to selection. Therefore, the main objective of this study was to identify the selection signatures related to immune response (IRE) and resilience to AD. . A total of 1,411 mink in five color types (black, demi, mahogany, pastel, and stardust) from the Canadian Centre for Fur Animal Research (Truro, Nova Scotia, Canada), which is an AD-positive facility, were genotyped using the Axiom Affymetrix Mink 70K single nucleotide polymorphism (SNP) panel. For the IRE trait, 264 mink were grouped into pairwise groups based on the combined results of counter-immunoelectrophoresis (CIEP) and enzyme-linked immunosorbent assay (ELISA) tests. Individuals in black (n = 29), demi (n = 131), mahogany (n = 76), and pastel (36) color types were also grouped based on combined results of CIEP and ELISA tests, respectively. For resilience traits, two methods were used to group CIEP-positive individuals into pairwise groups: 1) based on their general resilience performance (GRP, n = 30), which was measured by considering individual performance for feed conversion ratio, Kleiber ratio, and pelt quality, and 2) based on female reproductive performance (FRP, CIEP-positive dams only, n = 36), which was measured by the number of kits alive 24h after birth. The pairwise fixation index, nucleotide diversity, and cross-population extended haplotype homozygosity were used to detect selection signatures. Only SNPs detected by at least two methods were considered selection signatures candidates, including 619, 569, and 526 SNPs for IRE, GRP, and FRP traits, respectively. The genes annotated from the candidate SNPs were associated with previously reported traits influenced by AD, including immune system process, growth, reproduction, and pigmentation. Notably, two olfactory-related gene ontology (GO) terms were significant (q &amp;lt; 0.05) for all studied traits, suggesting AD might cause loss of smell in infected mink. Additionally, the differences in genes and GO terms detected across different color types for IRE indicated that mink of different color types may differ in immune response to AD. The mitogen-activated protein kinase (MAPK) signalling pathway was significant (q &amp;lt; 0.05) in Kyoto Encyclopedia of Genes and Genomes pathway analyses for FRP, indicating AD infection may disorder the MAPK signalling pathway, in turn influencing female reproductive performance. Our study has enhanced the understanding of genomic architecture underlying resilience of mink to AD and sheds light on the underlying biological mechanisms involved.

Seroprevalence of Viral Enzootic Diseases in Swine Backyard Farms in Serbia

Contrary to pig farming in developed Western countries, in a large part of the world, pigs are still traditionally kept in small backyard farms, usually for family needs. Their main characteristics are low biosecurity, swill feeding, natural breeding and uncontrolled trade. Given the high number of backyard farms in Serbia and the risk they are thought to pose to intensive pig farming, the main aim of this study was to evaluate the prevalence of major viral diseases of swine among traditionally kept pigs in small holdings with low biosecurity. For this investigation, 222 serum samples from 69 backyard holdings were randomly selected and tested for antibodies to Porcine Reproductive and Respiratory Syndrome virus (PRRSV), Aujeszky’s disease virus (ADV), Porcine Parvovirus (PPV) and Swine influenza Virus (SIV) by enzyme-linked immunosorbent assay (ELISA). The herd-level seroprevalence of PRRS, Aujeszky’s disease and PPV was 2.9%, 27.5% and 37.7%, respectively. Swine influenza seroconversion was not confirmed in any of the tested holdings. Despite widely distributed PPV and AD in backyard farms in Serbia, almost 50% of them are still negative for all the tested diseases. The backyard farms must be monitored, and owners must be educated as their role in eradication programs and obtaining country-free status may be crucial.

Natto extract inhibits infection caused by the Aujeszky's disease virus in mice.

Aujeszky's disease virus (ADV), also known as Suid alphaherpesvirus 1, which mainly infects swine, causes life-threatening neurological disorders. This disease is a serious global risk factor for economic losses in the swine industry. The development of new anti-ADV drugs is highly anticipated and required. Natto, a traditional Japanese fermented food made from soybeans, is a well-known health food. In our previous study, we confirmed that natto has the potential to inhibit viral infections by severe acute respiratory syndrome coronavirus 2 and bovine alphaherpesvirus 1 through their putative serine protease(s). In this study, we found that an agent(s) in natto functionally impaired ADV infection in cell culture assays. In addition, ADV treated with natto extract lost viral infectivity in the mice. We conducted an HPLC gel-filtration analysis of natto extract and molecular weight markers and confirmed that Fraction No. 10 had ADV-inactivating ability. Furthermore, the antiviral activity of Fraction No. 10 was inhibited by the serine protease inhibitor 4-(2-Aminoethyl) benzene sulfonyl fluoride hydrochloride (AEBSF). These results also suggest that Fraction No. 10, adjacent to the 12.5 kDa peak of the marker in natto extract, may inactivate ADV by proteolysis. Our findings provide new avenues of research for the prevention of Aujeszky's disease.

Meat Juice and Oral Fluid as Alternatives to Serum for Aujeszky Disease Monitoring in Pigs.

Aujeszky Disease Virus (ADV) is a double-stranded DNA virus with a lipoprotein envelope. The natural hosts of the infection are Suidae, but the virus can infect many other mammals. The gold-standard method identified by the WOAH for the diagnosis of Aujeszky disease is the ELISA method. The objective of this study was to compare the performance of meat juice and oral fluid matrices using a commercial ELISA kit designed for serum. A total of 80 blood and oral fluid samples were collected from four pig farms selected for this study. Diaphragm muscle samples of about 100 g and blood samples were collected from 213 animals at the abattoir. These biological matrices were collected from the same animals and tested using a competitive ELISA kit to detect antibodies against ADV. The relative accuracy of the meat juice compared to that of the serum was 96.7% (95% CI: 93.3-98.7%), with 206 correct results out of 213. The relative accuracy of the oral fluid compared to that of the serum was 61.3% (95% CI: 49.7-71.9%), with 58 correct results out of 80. Meat juice has a better combination of sensitivity and specificity than oral fluid. The usage of meat juice in routine diagnostic examinations could be achieved after further investigations to optimize the procedure.

Abortion and fetal death in sows.

Abortion in sows or the expulsion of foetuses between days 35 and 109 of gestation results in major financial losses. Abortion is the result of maternal failure due to factors interfering with either the endocrine control of pregnancy or causing endometrial damage. In addition, causes of fetal origin, due to infections with a special affinity for the fetus, can also lead to abortion. Many different non-infectious and infectious factors may be involved. Non-infectious risk factors include inappropriate ambient temperature, seasonal effects, different stress factors, and toxic substances. Microorganisms that may cause abortion can be classified as facultative pathogenic and specific pathogenic microorganisms. The first category includes mostly bacteria that are endemic in many pig farms and that are normally harmless commensals. They can cause abortion only in case of decreased immunity or other predisposing factors. Different specific pathogenic microorganisms, especially viruses and bacteria, can cause fetal death and abortion. Some may have a special affinity for the reproductive tract and the foetuses (e.g. Aujeszky's disease virus, parvovirus, Leptospira sp.), while others may cause clinical disease or fever in sows (e.g. swine influenza viruses, Erysipelothrix rhusiopathiae), subsequently leading to abortion. Diagnosis of fetal death and abortion is challenging due to the broad array of processes that may be involved, and the fact that the inciting cause may happen well in advance of the abortion. The diagnostic process should start with a thorough evaluation of the clinical problem and the farm. In case an infectious cause is suspected, proper samples should be collected, preferably from different sows and foetuses, for the detection of specific pathogens. Prevention of abortion mainly relies on the implementation of good management practices, hygiene and biosecurity measures, and for some pathogens also on vaccination.