Effects on ovine fetuses of exposure to ovine progressive pneumonia virus

Fetuses of 20 pregnant ewes at 4 gestational periods (45, 55, 85, and 100 days) were inoculated with ovine progressive pneumonia virus. Fourteen of 16 fetuses exposed to virus before gestational day 80 were either resorbed or expelled, whereas 10 of 15 fetuses exposed to virus after day 80 were normal at birth. Three of the 9 expelled fetuses and 1 of 2 newborn lambs had accumulations of lymphoid cells in the lungs. Virus was readily isolated from the tissues of expelled fetuses and newborn lambs. Lambs did not have precipitating antibody to the virus at birth, but 3 to 5 lambs had specific antiviral antibody at 18 months of age.     

Appearance of ovine progressive pneumonia virus outside the United States

A recently isolated Israeli retrovirus from a sheep with maedi-visna was compared with other retroviruses, using cDNA-RNA hybridization in solution. The Israeli isolate was shown to have close, if not identical, genetic homologic features with the ovine progressive pneumonia virus reported in the United States, rather than with those of the maedivisna viruses of European origin.     

Vasculitis associated with ovine progressive pneumonia virus infection in sheep

Vasculitis, involving small muscular arteries and arterioles, was found in 5 of 18 sheep naturally infected with ovine progressive pneumonia (OPP) virus and in 5 of 11 sheep experimentally infected with OPP virus. In order of frequency, arterial lesions were seen in carpal joint capsules, kidneys, meninges, brains, lungs, and tracheas. The lesions were intramural edema and hemorrhage, mononuclear cell infiltration, fibrinoid necrosis of media, and thrombosis. The vascular lesions were frequently associated with interstitial pneumonitis, arthritis, and encephalitis also induced by OPP virus.     

Mastitis associated with ovine progressive pneumonia virus infection in sheep

Eighteen ewes were experimentally infected with ovine progressive pneumonia virus and their mammary glands were examined for lesions and virus at 2.5 to 10 years postinoculation. Lesions were seen in 14 of 18 sheep; virus was isolated from 4 of 8 sheep. Lesion consisted of an interstitial accumulation of lymphocytes with periductal lymphoid nodules, and epithelial vacuolation and necrosis at the site of the lymphoid nodules.     

Arthritis associated with ovine progressive pneumonia

Chronic nonsuppurative arthritis developed in carpal and tarsal joints of border Leicester sheep that were naturally or experimentally infected with ovine progressive pneumonia (OPP) virus. Clinical signs of arthritis began at 1 to 6.2 years of age (1 to 5.7 years after inoculation of OPP virus) and slowly progressed in severity until the sheep died or were killed. The joint lesions were characterized by edema, hyperemia, hyperplasia, and necrosis of the synovial membrane; by necrosis and erosion of articular bone; by necrosis and fibrosis of subchondral bone; and by extensive periarticular fibrosis. The OPP virus alone was isolated from arthritic joints. In 7 of 14 sheep, the arthritis was associated with interstitial pneumonitis induced by the virus. Therefore, it was concluded that OPP virus was the cause of arthritis in this group of sheep.     

Minimum intravenous infectious dose of ovine progressive pneumonia virus (OPPV)

The minimum intravenous infectious dose for ovine progressive pneumonia virus (OPPV) WLC1 was determined using twenty-four 6 month-old lambs. Twelve groups of two 6 month-old lambs were inoculated intravenously (i.v.) with tissue culture fluid containing ovine progressive pneumonia virus (OPPV) WLC1 titers ranging from 10^7.6 TCID₅₀/lamb down to 10^−3.4 TCID₅₀/lamb and were monitored for seroconversion using the OPPV agar gel immunodiffusion assay (AGID). Fifteen of the 16 lambs given equal or greater than 10^0.6 TCID₅₀ seroconverted, and virus could be isolated from peripheral blood leukocytes in 13 out of the 15 of these lambs. None of the eight lambs receiving less than 10^0.6 TCID₅₀ seroconverted during the 12 months. The results of this study indicated that 10^0.6 or 4 TCID₅₀/lamb given i.v. was capable of establishing infection.     

Breed susceptibility to ovine progressive pneumonia (maedi/visna) virus

In this retrospective study of breed differences in susceptibility to disease caused by ovine progressive pneumonia (OPP) virus, 29 Border Leicester sheep were compared with 46 Columbia sheep. As judged by frequency and severity of clinical signs and lesions attributable to the infection, Border Leicester sheep were markedly more susceptible than Columbia sheep and experimentally infected sheep were slightly more susceptible than naturally infected sheep. Differences in susceptibility to infection by the virus were not determined.     

Diagnostic features of ovine progressive pneumonia.

Ovine progressive pneumonia, a chronic, insidious disease of adult sheep, has a relentless course leading to dyspnea, emaciation, and death. Clinical observations and serologic tests are adequate for making a tentative diagnosis. The agar gel immunodiffusion test seems to be the best serologic procedure for indicating infection with the virus but cannot be used to predict morphologic changes or clinical disease, inasmuch as many clinically unaffected animals carry the virus. A definitive diagnosis is based on finding lesions and isolating virus. Affected lungs are large and heavy as a result of interstitial accumulation of lymphoid cells and fibromuscular tissue. Frequently, interstitial lesions are accompanied by bronchopneumonia from secondary bacterial infection. The causal virus can be isolated from infected lungs by cocultivation with primary ovine or bovine cells.     

Immunodiffusion test for ovine progressive pneumonia.

An agar gel immunodiffusion test was developed to detect precipitating antibody against ovine progressive pneumonia (OPP) virus. The test was conducted in plastic petri dishes containing 6 ml of 1% purified agar in tris buffer and 8% sodium chloride. Wells for serum and antigen were 8 mm in diameter and were cut in a hexagonal pattern 3 cm from a central well. Tests were read at 24 and 48 hours. Soluble antigen for the test consisted of concentrated nutrient medium removed every 2 weeks from a cell culture persistently infected with isolate WLC 1 of OPP virus. Specificity of results was verified by testing serums from experimentally exposed sheep and appropriate controls. Two lines of precipitate formed with some serums from experimentally inoculated sheep. Serums taken soon after exposure of sheep to the virus and those taken 3 to 4 years after exposure frequently formed only 1 line of precipitate. Of 37 lambs inoculated with OPP virus, 25% of those tested were positive by postinoculation (PI) month 1, 79% of those tested were positive by PI month 3, and all of those tested were positive by PI month 6. The test appears adequate to detect exposure of sheep to OPP virus.     

Eradication of ovine progressive pneumonia from sheep flocks

Two management methods for controlling ovine progressive pneumonia in sheep were evaluated. By a test and cull method, wherein seropositive sheep were culled from the flock, the causative virus was eradicated from a closed flock and controlled in an open flock. By an isolate-and-test method, wherein lambs were removed from infected ewes before nursing and reared in isolation, the virus was eradicated in 1 of 2 attempts to establish virus-free flocks. It was concluded that either method should be effective in controlling ovine progressive pneumonia. Results indicated that annual serologic monitoring of a virus-free flock would be necessary to ensure that the virus-free status is maintained.     

Ultrastructure and frequency of mastitis caused by ovine progressive pneumonia virus infection in sheep

Twenty-five sheep, experimentally (n = 15) or naturally (n = 6) infected with ovine progressive pneumonia virus and noninfected controls (n = 4), were evaluated for histological and ultrastructural lesions of mastitis. Histologically, nine of 15 experimentally infected sheep and all six naturally infected sheep had lympho-plasmacytic mastitis. Severity of the lesion increased with length of time after infection. Periductal lymphatic nodules were seen in five sheep experimentally infected for 2.8 years or longer and in five naturally infected sheep that were 3.7 years old or older. Ultrastructurally, responses to ovine progressive pneumonia virus were diffuse lympho-plasmacytic infiltrates in glandular interstitium, lymphocytic and occasional plasmacytic infiltrates in ductal walls and lumens, lymphoblasts surrounded by small lymphocytes in glandular interstitium, and degeneration of epithelium releasing cells and cellular debris into the lumen. Based on the prevalence of lesions, the mammary tissue was more susceptible to ovine progressive pneumonia virus than other target organs: lung, brain, and synovium. Lesions did not differ between breeds of sheep. Ovine progressive pneumonia virus was not seen in the mammary tissue but was isolated from 15 of 17 mammary glands.     

Elevated concentrations in serum immunoglobulins due to infection by ovine progressive pneumonia virus.

Sixty-seven serum samples were obtained from 2 sheep flocks. Agar gel immunodiffusion (AGID) was used to separate progressive pneumonia virus (PPV)-infected sheep from noninfected sheep by the presence of precipitating antibodies. Immunoglobulin (Ig), total protein, and albumin concentrations were then measured from all 67 sera to determine whether differences existed between PPV-infected and non-infected sheep. A significant difference (P less than 0.0005) was found in both total protein and Ig concentration between PPV-infected and noninfected sheep. This corresponding difference was absent in albumin measurements. The significant differences (P less than 0.0005) in Ig and total protein concentrations were then used to evaluate a field test for diagnosing progressive pneumonia. The possibility of using either total protein or Ig concentrations as a field test was found to be highly unlikely due to variation in individual values.     

Microimmunodiffusion test for diagnosis of ovine progressive pneumonia.

Two microimmunodiffusion tests (MIDT) for detection and measurement of ovine progressive pneumonia antibody are described. Substrates of various salt concentrations and pH were used to determine the optimal conditions for the tests. In comparisons between two MIDT and one macroimmunodiffusion test, sera from cull ewes were used. The MIDT require less reagents and were more responsive than the macroimmunodiffusion test. After extended incubation of the test materials, results in all three tests were comparable.     

Early detection of maedi-visna (ovine progressive pneumonia) virus seroconversion in field sheep samples.

The aim of this work was to investigate whether an enzyme-linked immunosorbent assay (ELISA) was useful for early detection of maedi-visna virus (MVV) infection in sheep under field conditions. An ELISA based on p25 recombinant protein and a gp46 synthetic peptide was used. Sequentially obtained serum samples (n = 1,941) were studied for 4 years. ELISA results were compared with those of the agar gel immunodiffusion (AGID) test, and results of both tests were compared with a reference result established using consensus scores for at least 2 of 3 serologic techniques (AGID, ELISA, and western blotting, which was used to resolve result discrepancies between the other 2 techniques). A total of 247 discrepancies were observed between ELISA and AGID. Of these, 131 were due to an earlier detection of 120 sera by the ELISA and 11 sera by AGID. The remaining discrepancies (116) were due to the presence of false reactions in both tests. Fewer false-negative results were found by ELISA than with AGID (6 vs. 69 sera, respectively), whereas the number of false-positive results was virtually the same for ELISA and AGID (21 vs. 20, respectively). In relation to the reference result, ELISA sensitivity and specificity were 97.8% and 98.2%, respectively, whereas values for AGID were 76.3% and 98.3%, respectively. The agreement between ELISA and the reference result was higher than that between AGID and the reference result (K value: 0.96 and 0.77, respectively). A variation in the ELISA signal (based on optical density) was observed during the study period, suggesting different antibody levels throughout the animal's life. The ELISA was useful for detecting MVV-infected sheep in field conditions and has potential for use in control and eradication programs.     

Prevalence and effect of subclinical ovine progressive pneumonia virus infection on ewe wool and lamb production

The prevalence of infection with ovine progressive pneumonia (OPP) virus and its effects on ewe wool and lamb production were investigated in a flock of 2,976 ewes of 6 breed types (Rambouillet, Targhee, Columbia, Polypay, 1/4 cross Finnsheep, and 1/2 cross Finnsheep). Prevalence of seropositivity was significantly (P less than or equal to 0.01) lower among Rambouillet and Targhee breeds (44 and 42%, respectively), intermediate in Polypay, Columbia, and 1/4 cross Finnsheep (approximately 53%), and higher among 1/2 cross Finnsheep (62%). Seropositivity increased with age in all breed types from 11% at 1 year of age to 93% at greater than or equal to 7 years of age. Lateral disease transmission is indicated by linear increase of seropositivity prevalence with increasing age, including that in sheep greater than 6 years old. Subclinical infection with OPP virus had no apparent detrimental effect on number of lambs born, lamb viability, birth weight, number of lambs weaned, or growth rate of single and twin lambs, compared with findings for noninfected sheep in the same flock. Mature ewe body weight and grease fleece weight did not differ between subclinically infected seropositive and seronegative ewes. Subclinical infection with OPP virus does not appear to have an adverse economic effect on ewe wool and lamb production. Culling rate attributable to clinical manifestation of infection with OPP virus must be accurately determined before the true effects of virus infection on production can be determined and an eradication program can be recommended.     

Failure of experimental vaccines to protect against infection with ovine progressive pneumonia (maedi-visna) virus

Cell culture medium was harvested from cells infected with ovine progressive pneumonia (OPP) virus and used to prepare killed virus vaccines. Virus was inactivated by either heat, formalin, or ethyleneimine and used either without adjuvant, with Freund incomplete adjuvant, or with aluminum hydroxide adjuvant to vaccinate sheep. The sheep produced precipitating antibody against the virus but were not protected against infection when challenged with live OPP virus.     

Seroprevalence of ovine progressive pneumonia virus in various domestic and wild animal species, and species susceptibility to the virus

Ovine progressive pneumonia is caused by a lentivirus of known infectivity only for sheep and goats. Virus susceptibility of 11 other species of animals was examined. Species included cattle, chickens, deer, dogs, goats, hamsters, horses, mice, pigs, rabbits, and rats. Of these species, only goats and rabbits could be experimentally infected with the virus. The infection in rabbits was acute, and virus did not persist or induce antibody production as it does in sheep and goats. Sera obtained from several people working in close contact with the virus and from several wild species, with unknown exposure history, were tested for antibodies to viral antigens. All results were negative. Knowledge of the host range of this virus is important for scientific studies and for virus eradication programs.     

Serologic survey of prevalence of ovine progressive pneumonia in Idaho range sheep.

Blood samples from 2,310 mature sheep in 3 Idaho range flocks were examined by agar gel immunodiffusion to determine the prevalence of ovine progressive pneumonia. The prevalence ranged from 58% for all ages combined in one flock to 90% of cull ewes in another flock. Age-specific prevalence rates increased from 16% in yearlings to 83% in ewes greater than or equal to 7 years old. Rambouillet sheep had a significantly (P less than 0.01) lower prevalence than sheep of 5 other breeds, whereas one-half Finnsheep crosses had a significantly (P less than 0.01) higher prevalence than sheep of other breeds. Within breed and age, there was no significant difference in reproductive performance between seropositive and seronegative ewes.     

Seroprevalence of ovine progressive pneumonia virus in sheep in the United States as assessed by analyses of voluntarily submitted samples.

Ovine progressive pneumonia (OPP) is a lentivirus-induced disease of sheep in the United States that is similar, if not identical, to maedi/visna in many other countries. Prevalence estimates of seropositivity to this virus in sheep in the United States have been confined to limited groups or flocks of sheep and have varied from 1 to 90%. In this study of detection of antibodies against OPP virus, we found a lower general prevalence of antibodies to OPP virus in sheep than was previously reported. Of 16,827 sheep from 29 states in the United States, 26% were seropositive and 48% of 164 flocks that were tested had 1 or more seropositive sheep. Seropositivity to OPP virus for sheep within special categories was determined, although nonrandom samples that were available may have biased the results. Within regions of the United States, prevalence was highest in the Rocky Mountain region at 49% and lowest in the northern Atlantic region at 9%. Seropositive sheep were not evenly distributed among flocks, but were clustered in a few flocks of sheep. A high number of flocks had no or few seropositive sheep. Prevalence increased with age from 4% at less than 1 year to a plateau of 34% at 4 years. Seropositivity was variable among breeds and was not associated with sex, wool class, or place of origin of ancestors.