Observation on the remarkable stability of transmissible mink encephalopathy virus.

Transmissible mink encephalopathy (TME) virus retained biological activity after prolonged storage in formalin (about three and a half years) and in paraffin tissue blocks used for histologic preparations (about six and a half years). Thus, TME tissues in formalin and in paraffin blocks should be handled as contaminated materials.     

Experimental infection of Nigerian sheep and goats with bovine virus diarrhoea virus

Apart from leucopenia and a low grade pyrexia Nigerian goats showed no ill effects following inoculation with bovine virus diarrhoea virus. Sheep showed a variable leucopenia without pyrexia. No difficulty should arise in differentiating BVD infections from peste des petits ruminants.     

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.     

Experimental inoculation of raccoons (Procyon lotor) with Spiroplasma mirum and transmissible mink encephalopathy (TME)

The primary objective of this study was to determine whether or not Spiroplasma mirum would be capable of producing lesions of transmissible spongiform encephalopathy (TSE) when inoculated in raccoons (Procyon lotor) and, if that was possible, to compare the clinicopathological findings with those of transmissible mink encephalopathy (TME) in the same experimental model. For this purpose, 5 groups (n = 5) of raccoon kits were inoculated intracerebrally with either S. mirum and/or TME. Two other groups (n = 5) of raccoon kits served as sham-inoculated controls. All animals inoculated with TME, either alone or in combination, showed clinical signs of neurologic disorder and were euthanized within 6 mo post-inoculation (MPI). None of the carcasses revealed gross lesions. Spongiform encephalopathy was observed by light microscopy and the presence of abnormal disease-causing prion protein (PrP(d)) was detected by immunohistochemistry (IHC) and Western blot (WB) techniques in only the raccoons administered TME. Raccoons inoculated with Spiroplasma, but not administered TME agent, were euthanized at 30 MPI. They did not show clinical neurologic signs, their brains did not have lesions of spongiform encephalopathy, and their tissues were negative for S. mirum by polymerase chain reaction (PCR) and for PrP(d) by IHC and WB techniques. The results of this study indicate that Spiroplasma mirum does not induce TSE-like disease in raccoons.     

Experimental infection of pregnant goats with swine fever virus

Thirteen pregnant goats were inoculated intravenously with the ALD strain of virulent swine fever (SF) virus on Days 64-84 of gestation. Dams showed transient and mild viremia, and produced high serum neutralizing (SN) antibody after inoculation. Six inoculated dams were reared until parturition occurred and bore six apparently normal, one apparently normal but dead, one mummified and three edematous kids. Neutralizing antibody was demonstrated in the pre-colostral sera obtained from all normal kids, but no SF virus was isolated from any of them. The other seven dams were killed on post-inoculation days (PID) 5-61, and fetuses, placenta and amnion were tested for the virus and SN antibody. All fetuses of five dams examined within PID 40 were positive for SF virus, but negative for SN antibody. SF virus was also isolated from one of three fetuses examined on PID 61. Conversely, the other two fetuses examined on PID 61 were negative for SF virus, but positive for SN antibody. Furthermore, SF virus was isolated from the placenta and amnion of all the dams.     

Second passage of sheep scrapie and transmissible mink encephalopathy (TME) agents in raccoons (Procyon lotor)

To determine the transmissibility and pathogenicity of sheep scrapie and transmissible mink encephalopathy (TME) agents derived from raccoons (first passage), raccoon kits were inoculated intracerebrally with either TME (one source) or scrapie (two sources-each in separate groups of raccoons). Two uninoculated raccoon kits served as controls. All animals in the TME-inoculated group developed clinical signs of neurologic dysfunction and were euthanatized between postinoculation month (PIM) 6 and 8. Raccoons in the two scrapie-inoculated groups manifested similar clinical signs of disease, but such signs were observed much later and the animals were euthanized between PIM 12 and 18. Necropsy revealed no gross lesions in any of the raccoons. Spongiform encephalopathy was observed by use of light microscopy, and the presence of protease-resistant prion protein (PrPres) was detected by use of immunohistochemical (IHC) and Western blot analytic techniques. Results of IHC analysis indicated a distinct pattern of anatomic distribution of PrPres in the TME- and scrapie-inoculated raccoons. These findings confirm that TME and sheep scrapie are experimentally transmissible to raccoons and that the incubation periods and IHC distribution for both agents are distinct. Therefore, it may be possible to use raccoons for differentiating unknown transmissible spongiform encephalopathy (TSE) agents. Further studies, with regard to the incubation period and the pattern of PrPres deposition by use of IHC analysis in bovine spongiform encephalopathy and for other isolates of scrapie, chronic wasting disease, and TME in raccoons are needed before the model can be further characterized for differentiation of TSE agents.     

First and second cattle passage of transmissible mink encephalopathy by intracerebral inoculation

To compare clinicopathologic findings of transmissible mink encephalopathy (TME) with other transmissible spongiform encephalopathies (TSE, prion diseases) that have been shown to be experimentally transmissible to cattle (sheep scrapie and chronic wasting disease [CWD]), two groups of calves (n = 4 each) were intracerebrally inoculated with TME agents from two different sources (mink with TME and a steer with TME). Two uninoculated calves served as controls. Within 15.3 months postinoculation, all animals from both inoculated groups developed clinical signs of central nervous system (CNS) abnormality; their CNS tissues had microscopic spongiform encephalopathy (SE); and abnormal prion protein (PrP(res)) as detected in their CNS tissues by immunohistochemistry (IHC) and Western blot (WB) techniques. These findings demonstrate that intracerebrally inoculated cattle not only amplify TME PrP(res) but also develop clinical CNS signs and extensive lesions of SE. The latter has not been shown with other TSE agents (scrapie and CWD) similarly inoculated into cattle. The findings also suggest that the diagnostic techniques currently used for confirmation of bovine spongiform encephalopathy (BSE) would detect TME in cattle should it occur naturally. However, it would be a diagnostic challenge to differentiate TME in cattle from BSE by clinical signs, neuropathology, or the presence of PrP(res) by IHC and WB.     

Coronavirus infection in mink (Mustela vison). Serological evidence of infection with a coronavirus related to transmissible gastroenteritis virus and porcine epidemic diarrhea virus.

Antibodies to a transmissible gastroenteritis virus (TGEV)-related coronavirus have been demonstrated in mink sera by indirect immunofluorescence, peroxidase-linked antibody assays and immunoblotting. This is the first serological evidence of a specific coronavirus infection in mink. The putative mink coronavirus (MCV) seems to be widespread in the Danish mink population with a prevalence approaching 100%. Analysis by immunoblotting has shown that MCV is closely related to TGEV by the spike (S), matrix (M) and nucleoprotein (N) polypeptides. Furthermore, antibodies to MCV also cross-reacted with N and M polypeptides of porcine epidemic diarrhea virus (PEDV). Thus MCV may occupy an intermediate position between the TGEV group of coronavirus and PEDV. The possibility that MCV may be associated with syndromes of acute enteritis in preweaning mink is discussed.     

Transmission of transmissible mink encephalopathy to raccoons (Procyon lotor) by intracerebral inoculation.

To determine the transmissibility of transmissible mink encephalopathy (TME) agent to raccoons and to provide information about clinical course, lesions, and suitability of currently used diagnostic procedures for detection of transmissible spongiform encephalopathies (TSEs) in raccoons, 4 raccoon kits were inoculated intracerebrally with a brain suspension from mink experimentally infected with TME. One uninoculated raccoon kit served as a control. All 4 animals in the TME-inoculated group showed clinical signs of neurologic disorder and were euthanized between 21 and 23 weeks postinoculation (PI). Necropsy examinations revealed no gross lesions. Spongiform encephalopathy was observed by light microscopy, and the presence of protease-resistant prion protein (PrPres) was detected by immunohistochemistry and Western blot techniques. Scrapie-associated fibrils were observed by negative-stain electron microscopy in the brains of 3 of the 4 inoculated raccoons. These findings confirm that TME is experimentally transmissible to raccoons and that diagnostic techniques currently used for TSE in livestock detect prion protein in raccoon tissue. According to previously published data, the incubation period of sheep scrapie in raccoons is 2 years, whereas chronic wasting disease (CWD) had not shown transmission after 3 years of observation. Because incubation periods for the 3 US TSEs (scrapie, TME, and CWD) in raccoons appear to be markedly different, it may be possible to use raccoons for differentiating unknown TSE agents. Retrospective genotyping of raccoons using frozen spleens showed that the raccoon PrP gene is identical to the mink gene at codons 179 and 224. Further studies, such as the incubation periods of bovine spongiform encephalopathy and other isolates of scrapie, CWD, and TME in raccoons, are needed before the model can be further characterized for differentiation of TSE agents.     

Experimental infection of fetal and newborn Suffolk sheep with scrapie virus

Fetal (n = 21) and newborn (n = 7) Suffolk sheep were inoculated with scrapie virus isolated from other Suffolk sheep. Twenty fetuses, 76 to 109 days of gestational age, were inoculated IM in the neck through the uterine wall and were examined for virus 47 to 322 days later by mouse inoculation. Scrapie virus was not detected before 254 days of age; only traces of virus were detected in 3 of 7 lambs examined thereafter (2 at 254 days of age and 1 at 322 days of age). Virus was limited to the supra-pharyngeal, prescapular, and mesenteric lymph nodes. Seven lambs were inoculated into the palatine tonsils with scrapie virus as newborns (3 to 12 days old) and were examined for virus when they were 147 to 210 days old. Virus was not detected in the lymphoreticular tissues or terminal portion of ileum of any lamb. Failure to find scrapie virus in these lambs and in most lambs inoculated as fetuses might indicate few had became infected. However, if most lambs and fetuses had become infected, the long zero phase of the infection could have accounted for failure to find scrapie virus in many of them examined too soon after inoculation. The limited findings of this study indicate that efforts to demonstrate prenatal or neonatal transmission of scrapie by detecting virus are hampered by the slowness of its replication.     

Experimental infection of Mouflon-domestic sheep hybrids with caprine arthritis-encephalitis virus

OBJECTIVE: To determine whether monocyte-derived macrophages from Mouflon-domestic sheep hybrids (Ovis musimon X Ovis spp) were susceptible to productive infection with caprine arthritis-encephalitis virus (CAEV) in vitro and whether experimental inoculation of Mouflon-domestic sheep hybrids with a molecularly cloned CAEV would result in persistent infection. ANIMALS: 5 Mouflon hybrids. PROCEDURE: Macrophage monolayers were inoculated with virus in vitro. Three animals were inoculated with virus intratracheally. RESULTS: Productive replication of CAEV was demonstrated in monocyte-derived macrophages following in vitro and in vivo inoculation. Titer of infectious cytopathic CAEV produced by macrophages from the Mouflon hybrids was similar to titers produced by macrophages from an infected goat or by synovial membrane cells. Isolation of virus from monocyte-derived macrophages and use of a semiquantitative polymerase chain reaction assay to amplify a portion of the viral genome demonstrated persistent virus replication in all 3 inoculated animals. Two weeks after inoculation of sheep, approximately 1 of 5,000 monocytes was harboring the virus. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that Mouflon-domestic sheep hybrids are susceptible to infection with isolates of CAEV that cause infection in domestic small ruminants.     

Experimental infection of caprine arthritis-encephalitis virus in sheep

AIM: To describe outcomes for horses diagnosed with incomplete, non-displaced fractures of the frontal plane of the radial facet (INFR) of the third carpal bone (C3) treated by placement of a lag screw across the fracture under arthroscopic guidance.

METHODS: Horses (n=13) diagnosed with INFR and treated between December 1999 and January 2005 using a lag screw placed over the fracture were studied. For each case, five horses matched for sire, age and sex which were not known to have INFR were sought for comparison. Racing performance data were collected from a commercial online database. The racing performance of cases pre- and post-operatively, and of cases and matched horses in the post-operative period was compared.

RESULTS: Sixteen INFR were found in the 13 horses. Radiographic evidence of healed fracture lines 2–4 months after surgery was seen in 11/16 (69%) fractures; 11/13 (85%) cases raced again after a median recovery period of 292 (range 149–681) days. Treatment was considered successful in 9/13 (69%) cases, which were still in training or had been retired for reasons other than lameness localised to the middle carpal joint at the end of the study period. Just 6/13 (46%) cases had raced prior to injury. The racing ability pre- and post-operatively of five cases was compared, three (60%) of which performed better post-operatively than they had before. There was no significant difference in racing longevity or ability post-operatively between patients and matched (control) horses.

CONCLUSIONS: Post-operatively, there was little difference in the racing performance between horses diagnosed with INFR which had a lag screw placed across the fracture line and horses matched for sire, age and sex which were not known to have INFR.

CLINICAL RELEVANCE: Horses which were diagnosed with INFR of C3 and had a lag screw placed across the fracture had a good prognosis for future racing performance.