Molecular characterization of lentiviruses from goats from Poland based on gag gene sequence analysis

Caprine arthritis-encephalitis virus (CAEV) infection in goats is worldwide but with higher prevalence in industrialized countries. While positive serology of CAEV in Polish goats was reported there was no genetic study of this virus. In this study, we described the molecular characterization of lentiviruses isolated from seropositive goats from Poland. We cloned and sequenced a fragment from the gag gene covering part of the coding sequences for the matrix (MA) p17 and for the capsid (CA) p25 proteins. Resulting nucleotide sequences were aligned with those from other ovine/caprine lentivirus isolates. We present data showing that the sequences of most goat lentivirus isolates are closer to the prototypic CAEV-Co isolate, nevertheless from one goat we isolated a virus that is closer to the sheep Maedi Visna virus (MVV) isolate. This might indicate a recent cross-species infection from sheep to goat.     

Serological evidence of caprine arthritis–encephalitis virus (CAEV) infection in indigenous goats in the Sultanate of Oman

Caprine arthritis encephalitis (CAE) is a chronic debilitating disease of goats caused by a lentivirus responsible for economic losses as a result of a drop in milk production and weight loss. The objective of the study was to determine if indigenous goats from five different regions in the Sultanate of Oman exhibit serological evidence of exposure to CAEV using a competitive-inhibition ELISA technique. Blood samples were collected from slaughtered goats (N = 1,110) and from the National Serum Bank (n = 528). In total, 83 (5.1%) of screened samples were classed as seropositive. The results provide the first serological evidence for the presence of CAEV in Oman.     

Experimental infection of sheep by caprine arthritis-encephalitis virus and goats by progressive pneumonia virus

The lentiviruses, caprine arthritis-encephalitis virus (CAEV) and progressive pneumonia virus (PPV) of sheep, cause major diseases in their respective hosts; however, the infectivity of these viruses for closely related species has not been determined. Experiments were conducted to determine whether CAEV would infect sheep and whether PPV would infect goats. Upon inoculation with CAEV, lambs developed a nonsuppurative arthritis and antibody to CAEV, and the virus was isolated up to 4 months later. Exposure of 3 lambs to CAEV-infected adult goats did not lead to demonstrable infection after 18 months. Young goats inoculated with PPV replicated the virus and developed arthritis and antiviral antibody. These results demonstrate that these distinctly different lentiviruses may infect and cause diseases in species other than their accustomed host. Presently used techniques may not be effective in differentiating which lentivirus is responsible for infection of sheep and goats. Our results also indicate that mixing sheep and goats may adversely influence attempts to eradicate lentiviruses from these species.     

Goat uterine epithelial cells are susceptible to infection with Caprine Arthritis Encephalitis Virus (CAEV) in vivo

ABSTRACT: The aim of this study was to determine, using immunofluorescence and in situ hybridization, whether CAEV is capable of infecting goat uterine epithelial cells in vivo. Five CAEV seropositive goats confirmed as infected using double nested polymerase chain reaction (dnPCR) on leucocytes and on vaginal secretions were used as CAEV positive goats. Five CAEV-free goats were used as controls. Samples from the uterine horn were prepared for dnPCR, in situ hybridization, and immunofluorescence. The results from dnPCR confirmed the presence of CAEV proviral DNA in the uterine horn samples of infected goats whereas no CAEV proviral DNA was detected in samples taken from the uninfected control goats. The in situ hybridization probe was complementary to part of the CAEV gag gene and confirmed the presence of CAEV nucleic acids in uterine samples. The positively staining cells were seen concentrated in the mucosa of the lamina propria of uterine sections. Finally, laser confocal analysis of double p28/cytokeratin immunolabelled transverse sections of CAEV infected goat uterus, demonstrated that the virus was localized in glandular and epithelial cells. This study clearly demonstrates that goat uterine epithelial cells are susceptible to CAEV infection in vivo. This finding could help to further our understanding of the epidemiology of CAEV, and in particular the possibility of vertical transmission.     

Impact of natural sheep-goat transmission on detection and control of small ruminant lentivirus group C infections

Dissemination of small ruminant lentivirus (SRLV) infections in Norway is affected by the different control strategies used for maedi-visna virus (MVV) infections in sheep and caprine arthritis-encephalitis virus (CAEV) infections in goats. Here we investigated SRLV phylogenetic group variants in sheep. CAEV-like isolates, belonging to phylogenetic group C, were found among both seropositive sheep and goats in mixed flocks, in which sheep and goats are kept together. Intra-herd clustering confirmed that mixed flock animals were infected by the same virus variant, suggesting ongoing interspecies transmission. Few sheep flocks were found to be infected with the MVV-like phylogenetic group A. The apparent absence of SRLV group A type in goats is probably due to the MVV control programme and animal management practices. SRLV group C targets lungs and mammary glands in sheep, and induces typical SRLV pathological lesions. SRLV group C isolated from the sheep mammary glands suggested a productive infection and potential for transmission to offspring. SRLV group C was most prevalent among goats. A lower PCR sensitivity in seropositive sheep suggested a lower load of SRLV group C provirus in sheep than in goats. Higher genetic divergence of group C than in other SRLV groups and extensive heterogeneity among group C isolates in the matrix C-terminal region demonstrate the need for identifying conserved target regions when developing PCR protocols for SRLV detection. As sheep and goats may serve as reservoirs for all SRLV genogroup types, successful control programmes require inclusion of both species.     

Effects of infection with caprine arthritis-encephalitis virus on milk production in goats

A recently assembled commercial herd of Alpine goats was studied. Milk production criteria--305-day milk production (M), butter fat content (BF), and solids nonfat content (SNF)--and somatic cell counts (linear score) were monitored by Dairy Herd Improvement Association test records. Milk samples from all milking goats in the herd were obtained for bacteriologic culture for mastitis organisms on 2 occasions; the infection rate ascribed to major pathogens was 3%. In November 1985, serum specimens were obtained from 154 does in first lactation. Of these, 56 (36%) were seropositive for caprine arthritis-encephalitis (CAE) antibodies by agar gel immunodiffusion (AGID), 91 (59%) were seronegative, and serotest results for 7 (5%) were inconclusive. In December, 80 seronegative and 48 seropositive goats remained in the herd and had 305-day projections available. The median production values for seronegative goats (1,539.5 lb of M, 52 lb of BF, 46 lb of SNF) were higher than those for seropositive goats (1,446 lb of M, 45 lb of BF, 44.5 lb of SNF), but this difference was only significant (t test, P less than 0.05) for BF. Does were ranked by a formula that combined M, BF, and SNF, with a desired minimal daily herd average of 5 lb of M, 3% BF, and 3% SNF. A decision was made not to keep offspring from does of the lowest quartile before CAE test results were obtained. This group consisted of 13 of the 80 (16%) seronegative goats and 18 of the 48 (38%) seropositive goats. Thus, a positive CAE test result by AGID was associated (chi 2, P less than 0.01) with poor production.(ABSTRACT TRUNCATED AT 250 WORDS)     

Relationships between infection with caprine arthritis encephalitis virus, intramammary bacterial infection and somatic cell counts in dairy goats.

Somatic cell counts, the bacteriological condition of the milk and antibodies against caprine arthritis encephalitis virus (CAEV) were measured monthly throughout lactation in 121 lactating goats of the Murcia-Granada breed in four commercial dairy goat herds. The prevalence of bacterial intramammary infection was 5.6 per cent and the prevalence of CAEV infection was 20.6 per cent. An analysis of variance revealed a significant effect of herd, intramammary infection and the interaction between intramammary infection and CAEV on the somatic cell count. In udder halves free of intramammary infection, the somatic cell counts were significantly lower in seronegative goats than in seropositive goats (P<0.05), but the difference was not significant in udder halves persistently infected by bacteria. There was a significant increase in somatic cell counts due to bacterial intramammary infection (P<0.01) in the seronegative goats, but this effect was not present in the seropositive animals.     

Caprine retrovirus infection in New South Wales: virus isolations, clinical and histopathological findings and prevalence of antibody

Caprine arthritis-encephalitis virus (CAEV) was isolated by explant cultures of carpal synovial membranes and lung from 7 goats in New South Wales. These goats were clinically affected with the arthritic, neurologic, and pneumonic forms of CAEV infection either singly or in combination. CAEV antibody was detected by the gel immunodiffusion precipitin (GDP) test in 5 of the 7 goats. Serum samples from 2,708 goats, from 115 herds, were examined for CAEV antibody using the GDP test. Approximately one-third of the animals and 82% of the herds tested had CAEV antibody. The infection was common in all breeds of dairy goats with an indication of a significantly lower prevalence in the Saanen breed (24.4%) compared to Nubians, British Alpines and Toggenbergs (43.8%, 38.7% and 39.1% respectively). CAEV antibody was also demonstrated in 11 of 230 Angora goats. The infection was equally common in all age groups, with slightly higher prevalence in males (83 of 230, 36%) compared to females (648 of 2,232, 29%). Among seropositive animals 85% were clinically normal. Of 280 clinically affected goats tested only 42% had detectable antibody. One of 5 sheep that had been in contact with infected goats in one herd had CAEV serum antibody.     

Viral expression and leukocyte adhesion after in vitro infection of goat mammary gland cells with caprine arthritis-encephalitis virus

A characteristic lesion in goats infected by the lentivirus CAEV is mastitis with lymphoid hyperplasia. In order to investigate the mechanism of lesion formation, cultures highly enriched in microvascular endothelial cells, mature and immature luminal epithelial cells, fibroblasts and myoepithelial cells were established from goat mammary gland biopsies. Their susceptibility to in vitro infection with two distinct types of CAEV was investigated by PCR, antigen expression and cytopathy. The capacity of infected mammary gland cells to bind uninfected caprine leukocytes was determined by flow cytometry. All cell types tested were susceptible to CAEV infection in vitro, with different levels of sensitivity according to cell phenotype. Our results suggest that the limited extent of natural infection of mammary gland cells reflects a protective local immune response, and that the myoepithelial cell could act as a reservoir cell. After infection, the mature luminal cell acquires the capacity to bind leukocytes in vitro, which could indicate a facilitation of cellular interactions. The distinct reactions of the different cell types to CAEV infection may be correlated with events leading to progressive lesion development during the natural infection.     

绵羊慢病毒自然感染绵羊的硬化性淋巴细胞性乳腺炎

７头来自新疆南部某绵羊慢病毒（ＯｖＬＶ）感染的羊场的绵羊用于本研究。用琼脂凝胶免疫扩散检查绵羊血清中对绵羊进行性肺炎（ＯＰＰ）病毒（ＯＰＰＶ）的抗体，结果表明有６例呈阳性，１例阴性，抗体效价在３年中呈下降趋势。４例血清学阳性边菜羊和１例阴性和田羊有不同程度的硬化性（纤维性）淋巴细胞性乳腺炎，小叶内有不等的淋巴细胞浸润，导管周围无淋巴滤泡形成，小叶间大量纤维组织增生。７例的肺、脑、关节、血管均无ＯｖＬＶ性特异性病变。从血清学阳性羊的外周血白细胞中未分离到ＯｖＬＶ。     

A Labelled Avidin–Biotin ELISA to Detect Antibodies to Caprine Arthritis-encephalitis Virus in Goats' Sera

A labelled avidin–biotin ELISA (lab-ELISA) was developed and compared with indirect ELISA (i-ELISA) and agar-gel immunodiffusion assay (AGID) for its efficacy in detecting antibodies against caprine arthritis-encephalitis virus (CAEV) in goat sera. The enzyme immunoassays were standardized using 113 sera from CAEV-negative goat flocks. The tests were compared using the results from 339 serum samples. The lab-ELISA showed the greatest number of positive results (94/339) as compared with AGID (51) and i-ELISA (64). The comparison of the other two tests with the lab-ELISA showed an agreement of 87.3% with AGID and 90.6% with i-ELISA. The lab-ELISA may be useful for screening large populations for CAEV antibodies, in epidemiological surveys and in the control of caprine arthritis-encephalitis.     

Immunomodulation of caprine lentiviral infection by interleukin-16

Interleukin-16 (IL-16) is a proinflammatory cytokine produced by a variety of cells including lymphocytes, macrophages, mast cells, and eosinophils. We have shown in our previous studies increased expression of IL-16 mRNA and protein in caprine arthritis-encephalitis virus (CAEV)-infected goats blood. In this study, we determined the immunomodulatory effects of IL-16 in vitro using cells derived from CAEV infected and uninfected goats. Human recombinant IL-16 (rhIL-16) significantly increased chemotaxis of peripheral blood mononuclear cells (PBMCs) of both control and CAEV-infected goats. Pretreatment of PBMC with anti-goat CD4 monoclonal antibody inhibited IL-16-induced chemotaxis of PBMC of control and infected goats suggesting that IL-16 exerts its action in goats primarily by binding to CD4. The CAEV proviral DNA was less in caprine monocytes treated with rhIL-16 infected in vitro with CAEV. These data suggest inhibitory effect of IL-16 on viral integration. Flow cytometric studies indicated a trend toward IL-16-induced increased expression of lymphocyte activation markers. Combined with our previously reported data, these experiments suggest that increased IL-16 expression during CAEV infection may inhibit viral integration.     

Specificity of absorbed ELISA and agar gel immuno-diffusion tests for paratuberculosis in goats with observations about use of these tests in infected goats

OBJECTIVE: To determine the specificity of serological tests that are currently used in veterinary diagnostic laboratories in Australia for detection of Mycobacterium avium subsp paratuberculosis infection in goats. DESIGN: A laboratory study. PROCEDURE: Four tests were studied, comprising AGID with M. a. paratuberculosis antigen derived from cattle isolates of caprine or bovine origin, the EMAI caprine Johne's disease absorbed ELISA and the CSL PARACHEK Johne's absorbed EIA. The specificities of AGID and ELISA for paratuberculosis (Johne's disease) were estimated after examining a panel of 1000 serum samples collected from goats in Western Australia, a region free of paratuberculosis. In addition a comparison was made of test performance in a small number of paratuberculous goats from New South Wales using sera from two archival collections. RESULTS: The specificity of the AGID tests was 100% while the specificities of the two absorbed ELISA were 99.7 to 99.8% at appropriate positive-negative cut-offs. Based on testing the small sample of sera from infected goats, the absorbed ELISA tests detected about twice as many goats with Johne's disease as the AGID. Each test detected paratuberculous animals regardless of whether infection was caused by cattle or sheep strains of M. a. paratuberculosis. CONCLUSIONS: Both ELISA and AGID tests for paratuberculosis have high specificity and can be used in a market assurance program without risk of generating large proportions of false positive test results. However, the results suggested the ELISA is more sensitive for detection of infected goats and should be used in preference to the AGID. The two formats of ELISA evaluated in this study have similar characteristics and could be used in paratuberculosis control programs for the goat industries, but further data on sensitivity would increase confidence in their application.     

Caprine arthritis-encephalitis virus infection of goats in South Australia

Caprine arthritis-encephalitis virus (CAEV) infection of dairy goats was shown by virus isolation and serology to be widespread in South Australia. CAEV was isolated at necropsy from 24 of 27 dairy goats with swollen joints from 13 herds, and from 9 of 30 liver dairy goats in 7 herds. Virus was isolated most frequently from synovial membranes, and occasionally from mammary glands, mammary lymph nodes, choroid plexus, lungs, spleen, bone marrow, salivary glands, leucocytes, synovial fluid and milk. Antibody to CAEV was detected in the serum of 13 of 17 of the necropsied goats tested in a single-line gel diffusion test, and in another 3 retested with a modified double-line technique. Serum antibody was also demonstrated in 61 of 77 dairy goat herds, many with histories of arthritis. In 1984 to 1986 the annual number of serologically positive serums and proportions of the numbers tested were 134 (40%), 121 (45%) and 42 (18%), respectively. CAEV was isolated from leucocytes of 8 live goats in 6 of these herds. In fibre goats antibody was detected in the serum of 25 Angora and 19 crossbreds (0.1%) from the 33,279 Angora, 1,705 Cashmere, 8,715 crossbred and 904 feral goats tested.     

Risk factors associated with caprine arthritis-encephalitis virus infection in goats on California dairies.

Log-linear methodology was used to examine relations among caprine arthritis-encephalitis virus (CAEV) seroreactivity and host/management factors in a cross-sectional study of 2,826 goats on 13 California dairies. The CAEV serologic status was associated with age and feeding method (pasteurized/unpasteurized milk), but not with breed. Data from a prevalence survey of 321 goats from 2 additional dairies demonstrated very good fit of the selected log-linear model (P = 1.00), indicating that the model was very appropriate to describe the relations. Odds of seropositivity and odds ratios were generated by use of a logit model derived from the log-linear model. Goats raised by the unpasteurized feeding method were estimated to have been 3.3 times more likely to be CAEV-seropositive than goats fed by the pasteurized method, when adjusted for the effects of age. Goats aged 2, 3, 4, and 5 or greater years were estimated to have been 1.7, 2.6, 4.5, and 5.7 times, respectively, more likely to be CAEV-seropositive than were yearling goats when ratios were adjusted for pasteurization status. Breed, gender, and herd of origin were not associated with CAEV seroreactivity when the effects of other factors were considered. Estimated odds of CAEV seroreactivity and the associated odds ratios for combinations of factor levels are reported. In this study, the magnitude and direction of the associations among CAEV serologic status, age, and pasteurized feeding methods were demonstrated.     

Prevalence, spread and control of caprine arthritis-encephalitis virus in dairy goat herds in New South Wales

SUMMARY A study of the prevalence, spread and control of caprine arthritis-encephalitis virus (CAEV) in dairy goat herds in New South Wales (NSW) during 1986–1988 found that 56.8% of 1484 goats in 14 dairy herds were infected with CAEV. The prevalence of CAEV infection within most herds not implementing control measures increased during the study. At the end of the study, 59.7% of 1322 goats were infected. The prevalence of CAEV increased with age. Differences between breeds were less apparent. Within seven herds with a high standard of identification of goats, 149 of 812 goats seroconverted in an ELISA. Of these newly infected goats, 142 (95.3%) were > 1 yr of age and 96 (64.4%) were > 2 yr suggesting lateral spread of the virus. Most of the goats > 2 yr of age had been in the milking herd for a minimum of 3 to 6 months. The high seroconversion rate within the milking herd suggested that factors other than the ingestion of infected colostrum and milk before weaning were important for the spread of CAEV. Observations indicated that behaviour of goats, particularly reproductive behaviour among lactating does, and milking herd management practices are important in the spread of CAEV. A high density of livestock, poor livestock control and contamination of feed, water, equipment and personnel were implicated in transmission. Poorly functioning milking machines may also be involved. CAEV was eradicated from 3 herds by the implementation of strict control measures.     

Comparison of a maedi-visna virus CA-TM fusion protein ELISA with other assays for detecting sheep infected with North American ovine lentivirus strains.

A maedi-visna virus CA-TM fusion protein ELISA (MVV ELISA) was evaluated for the detection of antibody in sheep infected with North American ovine lentivirus (OvLV). The results of the MVV ELISA were compared with other assays for OvLV antibody and with viral infection in an intensively studied group of 38 sheep with a high prevalence of OvLV infection and disease. The sensitivity, specificity, and concordance of assays for OvLV antibody (MVV ELISA, indirect ELISA, Western blot, and AGID), virus (virus isolation, PCR, antigen ELISA), and OvLV-induced disease in each animal were compared with OvLV infection status as defined by a positive result in two or more of the assays. Five sheep met the criteria for absence of OvLV infection. The sensitivity of the MVV ELISA in detecting OvLV infected sheep was 64%, whereas the sensitivity of the other three tests for antibody ranged from 85 to 94%. All the antibody assays were 100% specific in this group of animals. Of the assays for virus, the PCR test had the highest sensitivity and the best concordance with OvLV infection, but it also had the lowest specificity of any of the virus or antibody assays. Among the antibody tests, the concordance of the MVV ELISA compared most favorably with the AGID test for detecting OvLV-infected sheep. Analysis of serum samples from 28 lambs experimentally-infected with one of three North American strains of OvLV suggested that there were no significant strain differences detectable by antibody assay. Twenty virus-inoculated lambs were positive by both the MVV ELISA and the AGID test, five lambs were MVV ELISA negative and AGID test positive, and three lambs were MVV ELISA positive and AGID test negative. No pre-inoculation samples were positive by either assay. In a longitudinal study involving seven lambs, antibodies to OvLV were detected by AGID 3-5 weeks post-inoculation, but were not detected by MVV ELISA until 5-10 weeks post-inoculation. Among 128 naturally and experimentally-infected sheep that were seropositive in the AGID test, the overall sensitivity of the MVV ELISA was higher in the naturally infected sheep (84%) than in the experimentally infected sheep (69%). The data indicated that the MVV ELISA represents a less sensitive, but specific alternative for the detection of OvLV antibodies.     

绵羊进行性肺炎病毒与山羊关节炎一脑炎病毒的血清学交叉反应的研究

本实验用琼脂凝胶免疫扩散试验（ＡＧＩＤＴ）和免疫印迹试验（ＩＢＡ）对实验感染绵羊进行性肺炎病毒（ＯＰＰＶ）的山羊血清与山羊关节炎—脑炎病毒（ＣＡＥＶ）抗原以及实验感染ＣＡＥＶ的绵羊血清与ＯＰＰＶ抗原的交叉反应进行了研究。４只接种ＯＰＰＶ的山羊中有一只山羊的血清可与ＣＡＥＶ琼扩抗原发生交叉反应,并在免疫印迹试验中可识别ＣＡＥＶ的ｇｐ４４、ｐ３５和ｐ２８。２只接种ＣＡＥＶ的绵羊中有一只绵羊的血清可与ＯＰＰＶ琼扩抗原发生交叉反应,并在免疫印迹试验中可识别ＯＰＰＶ的ｇｐ４４和ｐ２８。以上的交叉反应结果表明ＯＰＰＶ与ＣＡＥＶ的抗原之间具有密切的相关性,这对于ＯＰＰＶ通过山羊和ＣＡＥＶ通过绵羊的传代研究是非常重要的,并对将来的免疫预防策略具有重要的指导意义。     

PCR detection of lentiviral GAG segment DNA in the white blood cells of sheep and goats

A PCR assay for the detection of small ruminant lentiviral gag DNA (provirus) in the white blood cells of sheep and goats was developed and compared with a serological test (AGIDT). A sample of the DNA prepared from the white blood cells in 3 ml of blood from 208 sheep and goats from 18 different flocks was subjected to PCR assay. One of 85 animals from flocks accredited under the Dutch national MVV/CAEV control programme was positive by PCR while none was positive by AGIDT. In infected flocks, the AGIDT appeared slightly more sensitive, but preliminary results show that the sensitivity of the PCR assay may be further improved by increasing the number of monocytes tested. The PCR assay, however, was clearly more sensitive in detecting animals in the early stages of infection. With the use of a set of mixed primers and probes, the assay was able to detect the variety of CAEV and MVV strains occurring in the field.