Indurative lymphocytic mastitis in sheep after experimental infection with maedivisna virus

Summary

In order to provide further evidence for the association of an indurative lymphocytic mastitis in sheep with MVV (maedi‐visna virus) infection, an experimental study was performed. Fourteen MVV‐free pregnant ewes, 2 years of age, were divided into two groups. Eight were intravenously inoculated with MVV (strain ZZV‐1050); six ewes served as sham‐inoculated controls. Post‐mortem exat. inations were carried out at 8, 16 and 28 months.

After 8 months, the 3 infected ewes had indurated udders with extensive lymphoid proliferation around lactiferous ducts and in the acinar tissue. The ducts were often partially obliterated. After 16 months, one of the two infected ewes suffered from indurative lymphocytic mastitis. The other was free of specific udder lesions. After 28 months only one of three infected ewes had mild lymphocytic infiltration in the udder. None of the controls, two in each post‐mortem session, had lesions typical of this form of mastitis. The lesions were most severe 8 months after infection. At 16 and 28 months lesions were of a lesser degree or were absent.

The lung lesions in the infected ewes 8 months after inoculation were similar to the changes in the udder regarding the lymphoid accumulation, although the proliferation around bronchial tree and blood vessels was less pronounced. After 16 and 28 months all infected ewes had peribronchial and perivascular lymphocytic infiltration though of a lesser degree than after 8 months.

From these resutls it is concluded that in addition to the lung and brain lesions MVV infections may cause a specific indurative lymphocytic mastitis.     

Incidence of indurative lymphocytic mastitis in a flock of sheep infected with maedi-visna virus

Sheep in a flock in which 88 per cent of the ewes had antibodies to maedi-visna virus were clinically examined for udder induration during lactation and after drying off. On both occasions about half of the ewes had indurated udders. Histological examination revealed lymphocytic mastitis associated with maedi-visna virus infection, in the udders of six of 25 hoggs (24 per cent), 21 of 39 shearlings (53.8 per cent) and 42 of 67 ewes (62.7 per cent). Distinct lung lesions were found in 8 per cent of the hoggs, 12.5 per cent of the shearlings and 10 per cent of the ewes. The results of a clinical examination of dry udders were correlated with the histological findings.     

A chronic indurative mastitis in sheep,associated with maedi/visna virus infection

Summary

A possible association of a chronic indurative mastitis with mvv (maedi visna virus) infection in sheep was investigated. Sheep of four flocks (group A), in which insufficient lamb growth apparently associated with insufficient milk production and probably related to udder induration, was a serious problem, were clinically examined in mid‐lactation. The results were compared with four mvv‐free flocks (group B) without such complaints.

The incidence of udder induration in group A (n = 263) was about eight times higher than in group B(n = 206): 63.1 versus 8.0%. The clinical picture differed essentially between the groups. In group A the udder abnormalities were of a diffuse and indurative nature, involving both udder halves, while in group B the udder lesions were mainly nodular and often limited to one udder half.

Bacteriological examination revealed a difference in infection rate of the udders (6.8% in group A versus 14.1% in group B). A substantial difference was observed in a comparison of the bacteriological infection rate of the clinically abnormal udders (5.4% in group A, versus 47.0% in group B). Serological examination with an indirect ELISA revealed 81%, seropositive sheep in group A, versus 0% in group B. Twelve sheep of group A and five of group B, culled for reason of mastitis, were selected for pathological examination. The gross and microscopic lesions showed a pronounced difference between both groups. In group A a diffuse interstitial mastitis with slight to moderate fibrosis and a pronounced lymphoid hyperplasia was observed. In group B the mastitis had a nodular character, with a chronic galactophoritis, extensive fibrosis, and in some cases chronic abscesses. The lesions were often limited to one udder half Among the necropsied ewes of group A, nine showed lesions characteristic of maedi, whereas in group B no such lesions were observed.

The possible causal relation of this chronic indurative mastitis with mvv infection is discussed.     

A chronic indurative mastitis in sheep, associated with maedi/visna virus infection

A possible association of a chronic indurative mastitis with mvv (maedi visna virus) infection in sheep was investigated. Sheep of four flocks (group A), in which insufficient lamb growth apparently associated with insufficient milk production and probably related to udder induration, was a serious problem, were clinically examined in mid-lactation. The results were compared with four mvv-free flocks (group B) without such complaints. The incidence of udder induration in group A (n = 263) was about eight times higher than in group B (n = 206): 63.1 versus 8.0%. The clinical picture differed essentially between the groups. In group A the udder abnormalities were of a diffuse and indurative nature, involving both udder halves, while in group B the udder lesions were mainly nodular and often limited to one udder half. Bacteriological examination revealed a difference in infection rate of the udders (6.8% in group A versus 14.1% in group B). A substantial difference was observed in a comparison of the bacteriological infection rate of the clinically abnormal udders (5.4% in group A, versus 47.0% in group B). Serological examination with an indirect ELISA revealed 81% seropositive sheep in group A, versus 0% in group B. Twelve sheep of group A and five of group B, called for reason of mastitis, were selected for pathological examination. The gross and microscopic lesions showed a pronounced difference between both groups. In group A a diffuse interstitial mastitis with slight to moderate fibrosis and a pronounced lymphoid hyperplasia was observed. In group B the mastitis had a nodular character, with a chronic galactophoritis, extensive fibrosis, and in some cases chronic abscesses.(ABSTRACT TRUNCATED AT 250 WORDS)     

Bluetongue disease in Swiss sheep breeds: clinical signs after experimental infection with bluetongue virus serotype 8

Clinical disease of bluetongue (BT) in sheep may differ depending on breed, age and immunity of infected sheep and may also vary between serotype and strain of BT virus (BTV). Since there are no data available on the susceptibility of Swiss sheep breeds for BT, we performed experimental infection of the 4 most common Swiss sheep breeds and the highly susceptible Poll Dorset sheep with the BTV serotype 8 (BTV-8) circulating in Northern Europe since 2006. Clinical signs were assessed regarding severity, localisation, progression and time point of their appearance. The results clearly show that the Swiss sheep breeds investigated were susceptible to BTV-8 infection. They developed moderate, BT-characteristic symptoms, which were similar to those observed in Poll Dorset sheep. Regardless of breed, the majority of infected animals showed fever, swelling of the head as well as erosions of the mouth and subcutaneous haemorrhages.     

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.     

Pathogenicity of Mycoplasma capricolum in sheep after experimental infection

Mycoplasma (M.) capricolum has been frequently isolated from diseased as well as from healthy sheep in Morocco. In order to determine its pathogenicity for sheep, experiments were performed in three trials with the Moroccan isolate 012. The following results were obtained: Ewes inoculated intramammarily developed acute mastitis; the organism was transmitted to lambs suckling these ewes. Only a mild mastitis appeared after a second inoculum, performed 5 weeks after the first. Young lambs, 1 month of age, fed four times with M. capricolum broth culture, died during the septicemic phase, showing a generalized septicemia, polyarthritis and a diffuse interstitial pneumonia. Lambs, 2 and 3 months of age inoculated intrabronchially developed a disease, which appeared to be age and weight dependent. The more susceptible animals died within 1-2 weeks after infection. The older lambs recovered gradually from the disease, which was characterized by pneumonia, conjunctivitis and arthritis. A significant increase of antibodies against M. capricolum developed in the older animals in the complement fixation test.     

Semen changes in boars after experimental infection with pseudorabies virus

Two groups of adult boars were inoculated with a field strain of pseudorabies virus (PRV) by intranasal droplet; one group was given 5 x 10(5) median tissue culture infective doses (TCID50), and the other, 5 x 10(6) TCID50. (A third group was maintained as controls.) Ejaculates were examined twice a week for volume, sperm numbers, sperm morphology, and presence of PRV. Severe clinical disease with fever followed administration of the larger virus dose. Death (one boar), testicular degeneration, and transient elevation in spermatozoa with proximal cytoplasmic droplets were seen in different members of this group. The smaller dose resulted in seroconversion, but did not produce signs of disease. In this group, volume, sperm numbers, and sperm morphology did not decline when compared with base-line values or data of control animals. The virus was not isolated from semen. Effects of PRV infection on semen quality in boars seem to be related to the associated clinical signs of systemic disease.     

The role of virus dose in experimental bovine leukemia virus infection in sheep.

Twenty-four, six month old lambs were assembled into four groups of five animals each and one group of four animals. All groups were inoculated with lymphocytes from a single donor lamb infected with bovine leukemia virus. The inoculum varied from 250 to 250,000 lymphocytes, in tenfold increments. Animals were exposed by intradermal injection in the neck region immediately anterior to the left shoulder joint. All groups were monitored at 0, 3, 7 and 12 weeks after inoculation using the following procedures: a. Syncytia induction assay for detection of bovine leukemia virus in peripheral blood lymphocytes. b. Agar gel immunodiffusion against the gp51 antigen of bovine leukemia virus for the detection of antibovine leukemia virus gp51 antibody. c. Lymphocyte stimulation test for the assessment of cell-mediated immunity using mitogen, nonfractionated bovine leukemia virus antigen, and partially purified bovine lymphoma tumor-associated antigen for the in vitro activation of lymphocytes from bovine leukemia virus-inoculated and sham-inoculated, control animals. d. Routine hematological techniques for the assessment of total leukocyte and lymphocyte counts. The median infectious dose for lymphocytes from the single bovine leukemia virus-infected donor used in this study was determined to be 2000 cells. The syncytia induction assay detected more infected individuals (13/23) at an earlier time than did the agar gel immunodiffusion assay (10/23). Using either serological or virus isolation techniques, infected animals were first detected at three weeks postinoculation in the group receiving the high-dose inoculum and at seven weeks postinoculation in groups receiving low- or medium-dose inocula.(ABSTRACT TRUNCATED AT 250 WORDS)     

Observations on transplacental infection with bluetongue virus in sheep

Twenty-four ewes were inoculated with 1 of 2 strains of bluetongue virus type 4 at 40, 60, or 80 days of gestation. Two ewes aborted, 2 ewes died, and 1 was killed during the experiment, but their fetuses were recovered. At term, 2 mummified fetuses, 4 dead lambs, and 17 clinically healthy lambs were produced by 12 sheep, and the remaining 7 sheep were barren. Porencephaly and cerebellar dysgenesis were found in term lambs born to sheep inoculated at 40 and 60 days of gestation. Radiographic examination of 12 fetuses showed developmental ages far less than their chronologic age; 8 fetuses had skeletal growth-retardation lines, which were also observed in the dead lambs. A systemic lymphoreticular hyperplasia was observed in the dead lambs and in all lambs at 12 weeks of age; in 4 of the latter, granulomatous reactions were present in the liver and kidney. Lungs of the full-term lambs were reduced in weight and showed poor alveolar development and mononuclear cell infiltration, which persisted in the 12-week-old lambs. It was concluded that bluetongue virus is capable of causing not only gross abnormalities of the CNS, but also generalized growth retardation and fetal lymphoreticular hyperplasia.     

Further phenotypic characterization of target cells for bovine leukemia virus experimental infection in sheep

To determine the phenotype of target cells for bovine leukemia virus (BLV) infection in sheep, we analyzed blood lymphocytes from BLV-infected clinically healthy and leukemic sheep by use of monoclonal antibodies. In clinically healthy and leukemic sheep that were BLV-infected, the blood concentration of T lymphocytes was within normal values, but the number of B lymphocytes was increased in several cases. In addition, the number of blood lymphocytes expressing the BLV antigen correlated well with that of B lymphocytes. Double immunofluorescence staining demonstrated that lymphocytes expressing BLV antigens bore B-cell but not T-cell surface markers. Moreover, neoplastic cells in the lymph nodes of leukemic sheep were stained immunohistochemically with an anti-B monoclonal antibody but not with any of anti-T monoclonal antibody tested, indicating that tumor cells are of B-lymphocyte origin. Collectively, these results show that BLV antigen-positive cells obtained from BLV-infected sheep that have no clinical signs and BLV-induced lymphosarcoma cells belong to the B-lymphocyte lineage.     

Failure of foetal protection after vaccination against an experimental infection with bovine virus diarrhea virus

A protocol is described to measure the protection of the bovine fetus against an experimental bovine virus diarrhea virus (BVDV) infection after vaccination. Two inactivated experimental vaccines were applied twice with a 3 week interval. A mixture of three different Dutch field strains was used as challenge on mainly the 82nd day of gestation to vaccinated and unvaccinated control animals. The challenge was applied 5 months after completion of the two-fold vaccinations. All calves born from unvaccinated control animals were persistently infected. The calves born from dams vaccinated with the two different inactivated BVDV vaccines were persistently infected in 78 and 60%, respectively.     

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.     

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.     

Response of sheep to experimental concurrent infection with tick-borne fever (Cytoecetes phagocytophila) and louping-ill virus

The pathogenesis of concurrent Cytoecetes phagocytophila and louping-ill virus infection was studied in two experiments. In the first experiment 18 four- to seven-year-old rams were used. Ten were infected with C phagocytophila and five days later eight of these animals and the remaining eight sheep were infected with louping-ill virus. The two rams infected with C phagocytophila alone developed no clinical signs apart from a transient pyrexia, while only three of the eight rams infected with louping-ill virus alone showed mild clinical signs. In marked contrast, all eight dually infected sheep developed severe clinical signs with pronounced depression and dysentery and three died and five were killed in extremis. They developed higher titres of viraemia and the antibody response was depressed while necrotising lesions affecting a variety of organs were detected at post mortem examination. Rhizomucor pucillus was recovered from these lesions in seven of the eight sheep. A second experiment using 10 sheep, five aged seven months and five aged two to three years, confirmed the findings of the first experiment indicating that the age of the animal had not significantly influenced the initial result. It was concluded that C phagocytophila infection could enhance the pathogenicity of louping-ill virus and that, operating together, the two pathogens facilitated fungal invasion. It is postulated that sudden deaths in sheep recently transferred to tick-infested pastures may be due to this newly described syndrome.     

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.