Visna Research Articles

Retroviral coinfection (Jaagsiekte and Maedi-Visna viruses) in sheep with pulmonary tumors in Transylvania (Romania): retrospective study on 82 cases.

Ovine pulmonary adenocarcinoma (OPA) is an important viral-induced neoplasia in sheep caused by exogenous Jaagsiekte sheep retrovirus (exJSRV). Coinfection of exJSRV and Maedi-Visna virus (MVV) is reported in OPA cases, but its worldwide distribution and significance on lung pathology is not yet completely understood. This study aimed to investigate the MVV coinfection rate in 82 exJSRV-related OPA cases, and their pathological effects on lung parenchyma in slaughtered sheep in Transylvania (Romania). On gross examination, classical form of OPA was identified in 92.7%; no changes consisting with MVV interstitial pneumonia were identified in the included cases. The most common histological type of OPA was acinar (58.5%) and the myxoid growths were found in 18 cases. The exJSRV and MMV coinfection rate in examined sheep was 47.6% (39/82). The assessment of perineoplastic areas from coinfected animals, revealed interstitial lymphoplasmacytic infiltrates in all cases, lymphoid hyperplasia in 60.6% cases (20/33) and fibromuscular hyperplasia in 63.7% (21/33). This is the first report providing new data on distribution of OPA coexisting with MVV infection in slaughtered sheep in Romania. We consider that the OPA and MVV coinfection may play an important role on the severity of ovine chronic pulmonary diseases and further studies are needed to confirm this hypothesis.

Identification of concurrent infection with Jaagsiekte sheep retrovirus and maedi-visna virus in China.

Ovine pulmonary adenomatosis (OPA) and maedi-visna disease (MVD) are chronic and progressive infectious diseases in sheep caused by Jaagsiekte sheep retrovirus (JSRV) and maedi-visna virus (MVV), respectively. To investigate the pathological changes and conduct viral gene analysis of OPA and MVD co-occurrence in Inner Mongolia, China. Using gross pathology, histopathology, immunohistochemistry, ultrastructural pathology, PCR, and sequence analysis, we investigated the concurrent infection of JSRV and MVV in 319 Dorper rams slaughtered in a private slaughterhouse in Inner Mongolia, in 2022. Of the 319 rams included, 3 showed concurrent JSRV and MVV infection. Gross lung pathology showed diffuse enlargement, consolidation, and greyish-white miliary nodules on the lung surface; the trachea was filled with a white foamy fluid; hilar and mediastinal lymph nodes were significantly enlarged. Histopathology results revealed typical OPA and MVD lesions in the lung tissue. Immunohistochemical results were positive for JSRV envelope protein (Env) in the tumor cells and MVV CA in alveolar macrophages. Transmission electron microscopy showed several virions and autophagosomes in the lung tissue, severely damaged mitochondria, and the induced mitophagy. Nucleotide sequences obtained for JSRV env and MVV gag showed the highest homology with the Inner Mongolian strains of JSRV env (JQ837489) and MVV gag (MW248464). Our study confirmed that OPA and MVD co-occurrence and identified the pathological changes in Inner Mongolia, China, thereby providing references for the identification of concurrent JSRV and MVV infections.

Analysis of polymorphism in the TMEM154 gene associated with genetic resistance to Visna Maedi in semi - fine wool sheep breeds

Analysis of polymorphism in the TMEM154 gene associated with genetic resistance to Visna Maedi in semi - fine wool sheep breeds

Investigation of seroprevalence of small ruminant lentivirus infections in Erzurum province of Türkiye and determination of individual and environmental variables

Small ruminant lentiviruses (SRLVs) are chronic, incurable, and vaccine-free viral diseases that cause respiratory problems and nervous disorders and yield losses in sheep and goats. Caprine arthritis encephalitis virus in goats and maedi-visna virus in sheep have been named as SRLVs. This study aimed to determine the epidemiological status of SRLV infection in Erzurum province and to evaluate the risk factors of the disease based on breed, age, and sex. For this purpose, 204 animals including 184 sheep (Akkaraman, Morkaraman, and Hemşin breeds) and 20 goats (Anatolian Black goats) from 8 districts of Erzurum province (Aşkale, Hınıs, Horasan, Karaçoban, Palandöken, Pasinler, Pazaryolu, and Tekman) were included in the sample. Commercial antibody-ELISA kit was used to determine the seroprevalence of SRLV and 15.12% seropositivity was detected. In terms of SRLV, 14.67% of females and 20% of males were positive. In terms of breed, 20%, 13.76%, 0%, and 15% seropositivity was determined in Akkaraman, Morkaraman and Hemşin breed sheep and Anatolian Black goats, respectively. Although there was no statistically significant difference in terms of breed groups and sex, the detection rates in the districts were significant. In conclusion, the prevalence of SRLV infection was investigated in 8 locations of Erzurum province, which is one of the important centers of animal breeding and where small ruminant breeding is at a high level, and significant findings were obtained at the district level. With this study, updated data on seroprevalence of SRLV in the region were obtained and a broader perspective was tried to be provided by comparing with other SRLV studies in Türkiye and the world. These findings are important in terms of evaluating the prevalence and transmission risks of SRLV infections in the region and will shed light on future control and prevention strategies.

Newly developed peptide-ELISA successfully detected anti-IgG antibodies against Maedi-Visna virus in sheep

Maedi Visna Virus (MVV) is a retrovirus that can infect sheep. There is still no effective therapy or vaccine against this virus and timely diagnosis is important to combat the complications of the disease. In this study, we aimed to develop an ELISA using peptides derived from gag protein as antigen. For this purpose, B cell epitopes of gag protein were predicted and a docking analysis with the B cell receptor was performed to select peptides to be used in ELISA. After three soluble epitopes with the highest antigenicity were produced as peptides, the immunogenicity of each peptide was determined by ELISA using sheep serum samples categorized as MVV positive (n=24) and negative (n=13). Subsequently, in house ELISA using above mentioned immunogenic peptides as antigen was used to investigate MVV seroprevalence in sheep (n=88). According to the results, among three peptides, two of them strongly reacted with MVV positive serum samples and the mean absorbance values detected among positive and negative serum samples were statistically significant, indicating that these peptides were immunogenic (P=0.016 and P=0.038). The third peptide also reacted with positive serum samples but the mean absorbance value was not statistically significant and this peptide was considered non-immunogenic (P=0.175). The immunogenic two peptides showed the same high sensitivity and specificity values of 91.60 and 92.80 according to the commercial kit. Moreover, MVV seroprevalence detected by peptide-ELISAs using CKQGSKE and CRPQGKAGHKG peptides as antigen was 3.40 % and 4.5 %, respectively. As a result, it was shown that these peptides can be successfully used for serological diagnosis of MVV.

A high-density genome-wide approach reveals novel genetic markers linked to small ruminant lentivirus susceptibility in sheep.

Visna/Maedi virus (VMV) is lentiviral disease of sheep responsible for severe production losses. Multiple genomic regions associated with infection were reported indicating genetic complexity. In this study, a combined genome-wide approach using a high-density SNP array has been performed, comparing VMV-infected (n = 78) and non-infected (n = 66) individuals of the Valle del Belice breed. The serological tests showed a seroprevalence of 26%. The comparison among results from different approaches (GWAS, Fisher's exact test and the FST analysis) revealed two association signals: on OAR03 close to the GRIN2B gene and on OAR05 close to the TMEM232 gene. To the best of our knowledge, there has been no previous association between these genes and lentiviral infection in any species. The GRIN2B gene plays a role in pain response, synaptic transmission, and receptor clustering, while TMEM232 is involved in the development of immune-related disorders. The results highlighted new aspects of the genetic complexity related to the resistance/susceptibility to VMV in sheep, confirming that studies on different breeds can lead to different results. The ideal approach for validation of the markers identified in our study is to use samples from a population independent from the discovery population with the same phenotype used in the discovery stage.

Development of a Bead-Based Multiplex Fluorescent Immunoassay to Detect Antibodies against Maedi-Visna Virus in Sheep.

The Maedi-visna virus (MVV) causes a persistent infection in small ruminants, and its high genetic heterogeneity affects the performance of diagnostic tests when used in different populations. Therefore, the aim of this study was to develop a bead-based multiplex immunoassay tailored to detect antibodies against a Norwegian MVV strain. We used tissue samples from 14 PCR-positive sheep from a recent MVV outbreak in Norway to sequence the viral strain and produced recombinant antigens based on sequences from one animal. The assay included commercial TM-A and recombinant Norwegian p25, p16-25 and SU5 antigens. Cut-off values for each antigen were determined using receiver operating characteristic curves on 40 ELISA-negative and 67 ELISA-positive samples from the outbreak. The intraplate and interplate repeatability were investigated by testing a quadruplicate of five samples over three days, while the analytical sensitivity (aSe) and specificity (aSp) were measured in comparison to a commercial ELISA. The repeatability showed a coefficient of variation below 15% for most positive samples. The aSe was equal or higher for the multiplex assay than the ELISA, and the aSp of each antigen was 91.7, 93.3, 95.0 and 93.3% for p25, p16-25, SU5 and TM-A, respectively. The assay shows promising results; however, further evaluations of diagnostic characteristics are necessary before implementation in the Norwegian surveillance programme.

Phylogenetic analysis of small ruminant lentiviruses in Mongolian sheep supports an ancient east-west split for the genotype A.

The ovine maedi-visna virus (MVV) and caprine arthritis-encephalitis virus (CAEV) are small ruminant lentiviruses (SRLVs) with striking genetic and structural similarities. The presence of SRLV in Mongolian sheep and goats was serologically demonstrated more than a decade ago; however, the viral genotype remains unknown. In total, 329 blood samples were collected from two sheep breeds (i.e., Khalkha and Sumber) in Tov, Govisumber, Arkhangay, Dornogovi, Zavkhan, and Sukhbaatar provinces, Mongolia. Serological and phylogenetic analyses were performed regardless of any apparent clinical signs, although most of the animals appeared healthy. All sheep in three of the six provinces were seronegative, whereas the seroprevalence in the Tov, Govisumber, and Zavkhan provinces averaged 7.9%. Genomic DNA from seropositive animals was tested using hemi-nested polymerase chain reaction, and sub-genomic SRLV sequences were determined from nine samples. Mongolian SRLV sequences clustered within the divergent subtype A22, which was previously found only in Fertile Crescent regions, including Lebanon, Jordan, and Iran, where the first sheep-domestication (Ovis aries) occurred. According to the phylogenetic analysis, genotype A has two ancestors from the ancient Fertile Crescent: (1) Turkish strains and (2) Iranian, Jordanian, and Lebanese strains. The first ancestor spread westward, whereas the second spread eastward, ultimately reaching Mongolia.

Distribution of maedi-visna virus in different organs of the spontaneously affected small ruminants in India.

Maedi-visna (MV) is a small ruminant lentiviral (SRLV) disease affecting sheep and goats, and causes pathological alterations in various organs including lungs, pulmonary lymph nodes, mammary glands, joints, and CNS. Aims: Present study was focused to detect the MV virus (MVV) nucleic acid and MVV p28 antigen in different organs of the spontaneously MVV affected sheep and goats. Total of 657 samples were collected from sheep and goats (169 blood, 136 lungs, 96 pulmonary lymph nodes, 74 brain, 54 mammary gland, 78 joints, and 50 spleen) and screened for MVV nucleic acid using nested PCR assay. Serum samples were screened for SRLV antibodies by cELISA. Immunolocalization of MVV was demonstrated by using the polyclonal antibody against p28 antigen by immunohistochemistry in lungs, lymph nodes, mammary glands, and joint tissues. Out of 657 samples, 10.7% (70) were found positive for MVV. Among different organs, lungs showed highest positivity (25.7%) followed by mammary glands (14.8%), blood (9.5%), joint tissues (7.7%), brain (5.4%), and pulmonary lymph node (1.0%). SRLV antibodies were detected in 29.2% of the serum samples of both sheep and goats by cELISA. MVV p28 antigen immunostaining was observed in lungs, lymph nodes, mammary glands, and joint tissues. However, the presence of MVV p28 antigen could not be demonstrated in the brain tissues. The highest positivity of MVV in lung tissues indicated higher predilection of the virus in the pulmonary tissue.

Detection of Maedi Visna Virus (MVV) in milk samples of Indian goats

Detection of Maedi Visna Virus (MVV) in milk samples of Indian goats

Identification and characterization of linear epitopes of monoclonal antibodies against the capsid proteins of small ruminant lentiviruses.

Maedi-visna virus (MVV) and caprine arthritis encephalitis virus (CAEV) are members of a group of genetically highly homologous lentiviruses collectively referred to as small ruminant lentiviruses (SRLVs). SRLVs can infect sheep, goats and other small ruminants, causing multisystemic disease with progressive and persistent inflammatory changes, severely reducing animal productivity and impeding animal trade. The capsid protein of SRLVs, p28, is highly conserved among strains and is a commonly used marker for the detection of SRLVs. In this study, two monoclonal antibodies (mAbs), designated G8F7 and A10C12, against p28 were generated using a recombinant p28 protein expressed in Escherichia coli as an immunogen. Functional analysis showed that these two monoclonal antibodies could be used in iELISA, immunofluorescence assays (IFA) and western blot assays to detect p28 or Gag precursor proteins of SRLVs. Two linear epitopes, 61GNRAQKELIQGKLNEEA77 (E61-77) and 187CQKQMDRVLGTRVQQATVEEKMQACR212 (E187-212), which are recognized by G8F7 and A10C12, respectively, were identified through truncation of the GST-fused p28. Amino acid sequence alignment showed that the epitope E61-77 is conserved among SRLVs, with a dominant mutation site (K72R) that does not disrupt recognition by G8F7. E187-212 was found to exhibit variability among SRLVs, but the majority of mutant epitopes are recognized by A10C12, with the exception of a mutant epitope from an isolate with undefined subtypes from Ovis aries, which was not recognized. These findings may facilitate future study of SRLVs and promote the development of methods for the detection of these viruses.

The genetic variability of small-ruminant lentiviruses and its impact on tropism, the development of diagnostic tests and vaccines and the effectiveness of control programmes.

Maedi-visna virus and caprine arthritis encephalitis virus are two closely related lentiviruses which cause multisystemic, progressive and persistent infection in goats and sheep. Because these viruses frequently cross the species barrier, they are considered to be one genetic group called small-ruminant lentiviruses (SRLV). They have in vivo tropism mainly for monocytes and macrophages and organ tropism with unknown mechanisms. Typical clinical signs are pneumonia in sheep, arthritis in goats, and mastitis in both species. Infection with SRLV cannot currently be treated or prevented, and control programmes are the only approaches to avoiding its spread. These programmes rely mainly on annual serological testing and elimination of positive animals. However, the high genetic and antigenic variability of SRLV complicate their early and definitive diagnosis. The objective of this review is to summarise the current knowledge of SRLV genetic variation and its implications for tropism, the development of diagnostic tests and vaccines and the effectiveness of control and eradication programmes. Subject literature was selected from the PubMed and the Google Scholar databases. The high genetic diversity of SRLV affects the performance of diagnostic tools and therefore control programmes. For the early and definitive diagnosis of SRLV infection, a combination of serological and molecular tests is suggested. Testing by PCR can also be considered for sub-yearling animals. There are still significant gaps in our knowledge of the epidemiology, immunology and biology of SRLV and their impact on animal production and welfare. This information may aid selection of the most effective SRLV spread reduction measures.

Challenges using serological diagnostics in elimination of visna/maedi: Serological results from two outbreaks in Norwegian sheep

Detection of visna/maedi virus (VMV) infection is challenging, as no perfect reference test exists. Several factors can lead to false positive or negative results, including slow seroconversion, fluctuating antibody levels, virus heterogeneity and cross-reactive substances in serum. Norway aims to eliminate VMV, therefore a VMV flock diagnosis will have severe consequences for the farmer. The last two outbreaks occurred in the same geographical area in 2002–2005 and 2019–2020, including 61 and 9 flocks, respectively. The viral strains detected were genetically similar, indicating continuous, undetected VMV circulation in the sheep population. In this descriptive study, we investigate the challenges of correctly diagnosing VMV infection in sheep when the goal is elimination, and describe the distribution of infection within flocks. The choice of test influenced the number of false reactions, as only 40.6 % of the samples that tested positive by the screening ELISA used in 2019–2020 tested positive by two other commercial ELISA tests. In order to minimize the number of false positive VMV flock diagnoses, we investigated if the interpretation could be improved by resampling animals with ambiguous results. Only 26 % of the animals in negative flocks retrieved agreement between the two ELISAs at resampling. It is therefore debatable whether resampling is recommended. Further, we found a higher seroprevalence in older animals, and 36 of 95 offspring obtained different test results than their mother, supporting previous knowledge on slow seroconversion. Different ELISA tests were used in the two outbreaks. Some of the negative samples from 2002 to 2005 were positive when retested with the screening test used in 2019–2020. Hence, animals with false negative results in the 2002–2005 outbreak might be a reason for the continuous circulation of the virus.

147 Determining Status of Five Genetic Conditions in U.S. Sheep with a Medium-Density Ovine Bead Array

Abstract Using molecular information in genetic evaluation has become routine in most livestock species. Small ruminants, however, are lagging in use of this technology, partly due to the higher cost of genotyping versus the value of the individual animal. Using a single genotyping platform to simultaneously obtain information for making genomic predictions, identifying genetic condition status, and verifying parentage avoids costs of running multiple DNA tests. Our objective was to develop and validate a process to obtain accurate genotypes for genetic conditions using a medium-density array, and to estimate their frequencies in U.S. sheep breeds. Samples (DNA) from 10,569 sheep from 9 breeds in the National Sheep Improvement Program, primarily Katahdin (8,657), Rambouillet (886), and Polypay (584), were genotyped with a commercial 50k single nucleotide polymorphism (SNP) bead chip. Genotypes for 5 genetic conditions, ovine progressive pneumonia (OPP) susceptibility (TMEM154), scrapie susceptibility (PRNP), double muscle (MSTN), callipyge (CLPG), and booroola FecB (BMPR1B), were determined using 66 SNP genotypes extracted from the bead chip. Targeted SNP markers were replicated 2 to 8 times on the array. The TMEM154 diplotypes were obtained by combining genotypes at 10 SNP markers. Variants for PRNP codon 136 and 171 were determined using 6 markers. The accuracy of assigning genetic status was validated using 15 reference DNA with known genotypes submitted blindly to the commercial laboratory. Consistency of replicated calls for an SNP was also evaluated. Following in-house laboratory-based quality control of the assay, acceptance of a genotype involved two steps. First, at least 60% of the replicated SNP on an animal needed to be assigned a genotype. Second, amongst those SNP genotyped, at least 60% needed to detect the same nucleotide. This strategy resulted in 98.4% and 90.9% of the animals having genetic status scored for PRNP and TMEM154, respectively. Where genotypes were accepted, OPP and scrapie susceptibility was assigned based on TMEM154 diplotypes and PRNP codons 136 and 171 genotypes, respectively. As shown in Table 1, genetic condition status differed appreciably among Katahdin, Polypay, and Rambouillet. Nearly 60% of Katahdins were characterized as highly susceptible to OPP while such was so in less than 15% Polypay and Rambouillet. Most (93.3%) Polypay were deemed scrapie resistant, with a majority of Katahdin (93.1%) and Rambouillet (80.6%) either resistant or rarely susceptible to this disease. However, 18.6% of Rambouillet were characterized as highly susceptible to scrapie. No animals carried the callipyge or FecB mutation. The double muscle mutation also was rare; in Katahdins, 2.7 and 0.2% of animals carried one or two copies, respectively. The approach developed accounted for genotyping inaccuracies, with full validation of genetic status. Furthermore, considerable variation in susceptibilities to OPP and scrapie was detected among breeds, which can be used in defining breeding objectives.

In silico discovery of epitopes of gag and env proteins for the development of a multi-epitope vaccine candidate against Maedi Visna Virus using reverse vaccinology approach

Maedi Visna Virus (MVV) causes a chronic viral disease in sheep. Since there is no specific therapeutic drug that targets MVV, development of a vaccine against the MVV is inevitable. This study aimed to analyze the gag and env proteins as vaccine candidate proteins and to identify epitopes in these proteins. In addition, it was aimed to construct a multi-epitope vaccine candidate. According to the obtained results, the gag protein was detected to be more conserved and had a higher antigenicity value. Also, the number of alpha helix in the secondary structure was higher and transmembrane helices were not detected. Although many B cell and MHC-I/II epitopes were predicted, only 19 of them were detected to have the properties of antigenic, non-allergenic, non-toxic, soluble, and non-hemolytic. Of these epitopes, five were remarkable due to having the highest antigenicity value. However, the final multi-epitope vaccine was constructed with 19 epitopes. A strong affinity was shown between the final multi-epitope vaccine and TLR-2/4. In conclusion, the gag protein was a better antigen. However, both proteins had epitopes with high antigenicity value. Also, the final multi-epitope vaccine construct had a potential to be used as a peptide vaccine due to its immuno-informatics results.

Seroepidemiology of Maedi-Visna in Intensively Reared Dairy Sheep: A Two-Year Prospective Study.

The objective of this study is to prospectively evaluate the seroepidemiology of maedi-visna (MV) infections in intensively reared dairy sheep. A total of 407 purebred Chios and Lacaune ewes from four farms were surveyed for two consecutive years and were serologically tested semiannually with an indirect ELISA at pre-mating and pre-lambing. The farms' structure and management practices were similar and animal traits (age, breed, and production stage) were recorded. Based on the serological status, morbidity frequency measures were estimated, and ewes were categorized as constantly seronegative, constantly seropositive, seroconverted, seroreverted, or as animals with an intermittent presence of antibodies. During the study, period seroprevalence, incidence rate, and cumulative incidence were 84.8%, 33.6 new cases per 100 sheep-semesters, and 64.2%. Point-seroprevalence ranged from 48.5% to 96.0% among the studied farms and sampling occasions, and they increased by age. Increased morbidity frequency measures indicate the significance of horizontal transmission in intensive dairy sheep farms. A remarkable percentage of infected animals seroreverted (8.1%) or presented an intermittent presence of antibodies (10.3%) during the study, confirming the risk of misdiagnosis in cross-sectional studies and in the currently implemented testing and elimination programs. The serological patterns observed in our study need to be considered when studying MV epidemiology and for the designing of efficient MV elimination programs.

Degree of technification of the sheep herd may be associated with seropositivity for the maedi-visna virus In South Brasil

Maedi-Visna (MV) is a chronic progressive multisystem disease that may be asymptomatic for several months or years, but progress rapidly, and may result in death, when signs and symptoms evolve. Viral elimination occurs mainly through direct contact with positive animal secretions. There is no vaccine or treatment, and prophylaxis is necessary for the health of the herd. The present study aimed to verify the seropositivity of MV and evaluate the factors associated with the risk in sheep herds in Paraná. A total of 1549 serum samples were collected from 90 properties. An epidemiological questionnaire was applied to each property, and the variables were analyzed using the Epi-info program and R environment. Of the 1549 samples analyzed, 22 were positive (1.4%) for the micro-AGID test in 13.3% of the properties. Our study demonstrated variables associated with the prevention and the risk of seropositivity to MVV. Conducting a breeding season, supplying concentrated feed, and separating the breeding stock before birth were factors associated with protection, whereas the previous occurrence of problems with lice, breeding on pasture, and keeping cats close to the flock were factors associated with risk. The seropositivity observed in the present study suggests the circulation of MVV in sheep herds in Paraná, which reinforces the need to implement prevention and control measures since the level of technification may be associated with the occurrence of anti-MVV antibodies in herds.

O-116 Development of a multiplex immunoassay to detect antibodies against maedi-visna virus in sheep

O-116 Development of a multiplex immunoassay to detect antibodies against maedi-visna virus in sheep

Associations between small ruminant lentivirus infection and total milk yield and somatic cell count in a dairy sheep flock.

Small ruminant lentiviruses (SRLVs) are lentiviruses of sheep and goats, formerly known as maedi-visna (MV) in sheep and caprine encephalitis and arthritis in goats. In sheep, SRLVs commonly cause progressive pneumonia, wasting and indurative mastitis. SRLVs have a long latent period, and chronic production losses are often not recognised until very late. Few studies quantifying the production losses in ewes have been published, and none have been published under UK flock husbandry conditions. Production records of milk yield and somatic cell count (SCC) from a dairy flock of 319 milking East Friesian × Lacaune ewes identified as MV infected via routine serological screening for SRLV antibodies were used in multivariable linear regression modelling to estimate the impact of SRLV status on total milk yield and SCC. Milk yield was reduced in seropositive ewes by 8.1%-9.2%over an entire lactation. SCC counts were not significantly different in SRLV-infected andunifectedanimals. Further parameters, such as body condition score or clinical mastitis, that were not available may have clarified the underlying cause of milk yield drop. The study demonstrates substantial production losses in anSRLV-affected flock and highlightsthe impact of the virus on afarm'seconomic viability.

A Combined Approach for Detection of Ovine Small Ruminant Retrovirus Co-Infections.

Jaagsiekte retrovirus (JSRV)-induced ovine pulmonary adenocarcinoma (OPA) is an important ovine respiratory disease in Switzerland. Furthermore, ovine lungs with OPA frequently exhibited lesions suggestive of maedi-visna virus (MVV) or caprine arthritis encephalitis virus (CAEV) infection, indicating that co-morbidities might occur. Lungs and pulmonary lymph nodes were sampled from suspected OPA cases, inflammatory lung lesions and control lungs (total of 110 cases). Tissues were (a) processed for histology and immunohistochemistry (IHC), and (b) underwent DNA extraction and real-time PCR for JSRV, MVV and CAEV. Peptide sequences were used to generate virus-specific customized polyclonal antibodies. PCR-positive OPA cases and formalin-fixed and paraffin-embedded MVV- and CAEV-infected synovial cell pellets served as positive controls. Fifty-two lungs were histologically diagnosed with OPA. Histological evidence of MVV/CAEV infection was detected in 25 lungs. JSRV was detected by PCR in 84% of the suspected OPA cases; six were co-infected with MVV and one with CAEV. MVV was detected by PCR in 14 cases, and four lungs were positive for CAEV. Three lungs had MVV/CAEV co-infection. In IHC, JSRV was detected in 91% of the PCR-positive cases, whereas MVV and CAEV immunoreactivity was seen in all PCR-positive lungs. Although PCR showed a higher sensitivity compared to IHC, the combined approach allows for investigations on viral cell tropism and pathogenic processes in co-morbidities, including their potential interdependency. Furthermore, an immunohistochemical tool for specific differentiation of MVV and/or CAEV infection was implemented.