Retrovirus-associated ovine pulmonary carcinoma (sheep pulmonary adenomatosis) and lymphoid interstitial pneumonia. I. Lesion development and age susceptibility

To determine the lesion development of retrovirus-induced ovine pulmonary carcinoma (OPC), ten neonatal lambs were inoculated intratracheally with either 1) lung fluid preparations derived from a sheep with Type D retrovirus-associated OPC and concurrent ovine lentivirus (OvLV)-associated lymphoid interstitial pneumonia (LIP) (n = 8); or 2) lung fluid from a sheep with only OvLV-LIP (n = 2). Seven of eight neonates that received Type D retrovirus-associated OPC/OvLV-LIP lung fluid developed both OPC and LIP lesions between 9 and 32 weeks after inoculation. Mild OPC lesions consisted of foci of type II alveolar epithelial cells lining alveoli surrounded by minimal alveolar macrophage infiltrates. More severe OPC lesions consisted of multifocal aggregates of cuboidal to columnar neoplastic cells forming acini or masses associated with abundant alveolar macrophage infiltrates. Lesions of LIP consisted of peribronchiolar and perivascular lymphoid hyperplasia and heterogeneous interstitial leukocytic infiltrates. The two neonates that received OvLV-LIP lung fluid developed rapid and severe LIP, but not OPC lesions. Two lambs (inoculated as neonates with virus-free lung fluid) and three lambs (uninoculated contacts) served as controls and did not develop OPC. To investigate age susceptibility for development of OPC, 20 additional lambs within defined age groups (neonates, 2 weeks old, 5 weeks old, and 10 weeks old) received ultracentrifuged tumor homogenate. Neonatal to 5-week-old lambs inoculated with Type D retrovirus-associated OPC/OvLV-LIP tumor homogenate were equally likely to develop OPC, but lambs inoculated at 10 weeks of age were more refractory to tumor development.(ABSTRACT TRUNCATED AT 250 WORDS)     

A MUCOSAL DISEASE VIRUS AS A CAUSE OF ABORTION, HAIRY BIRTH COAT AND UNTHRIFTINESS IN SHEEP

A condition in Australian sheep resembling border disease was transmitted by the inoculation of pregnant ewes with material from affected lambs. This material contained mucosal disease virus (MDV). Twenty-two lambs comprising 6 from uninoculated control ewes, together with 11 with hairy coats and 5 with normal coats from inoculated ewes, were observed from 7 to 182 days after birth. Nine of the lambs from inoculated ewes died during the experiment from a variety of causes. Glial cell abnormalities were observed in control and affected lambs, but only 4 of the 11 hairy lambs were judged to have abnormal glial cells. There were no consistent histopathological findings indicative of MDV infection. MDV was recovered from tissues of all 11 hairy lambs, but not from any of the lambs with normal coats. The hairy lambs appeared to be immunologically tolerant to the virus. Susceptible sheep in contact with the hairy lambs were infected with MDV. It is suggested that a condition in Australian lambs characterised by hairiness of the birth coat and poor viability is due to foetal infection with a mucosal disease virus.     

Clinical and Para-clinical Findings of a Recent Outbreaks of Peste des Petits Ruminants in Iran

Peste des petits ruminants (PPR) is a highly contagious and infectious viral disease of domestic and wild small ruminants characterized by fever, erosive stomatitis, conjunctivitis, gastroenteritis and pneumonia. Goats are usually more severely affected than sheep. Peste des petits ruminants is caused by a paramyxovirus of the Morbillivirus genus. In March 2004, a flock of sheep in Tehran province with 430 deaths was visited. According to the history taken from the owner, at disease onset most of the deaths were recorded from adult sheep, 3 weeks later lambs (2 weeks to 4 months of age) showed the highest death rate. All animals from 3 months age received rinderpest vaccine 1 month after onset. Many of the lambs died just a few hours after their first sucking of the colostrum from infected mothers. Most of them showed very acute form of disease and died a few hours after onset of clinical signs. In clinical examinations most of the cases showed severe depression, high fever (41 degrees C), anorexia, mocopulurent nasal discharge, erosive and necrotic stomatitis (dental path, hard palate and cheeks), diarrhoea and dehydration. Para-clinical findings including histopathological, serological and haematological examinations also confirmed the presence of PPR in this flock. PPR outbreaks have been frequent in Iran in recent years. Further, we suggest that PPR is not a recent invader of Iran. The main difference in clinical signs between this outbreak and the same in other reports is that goats did not show any obvious signs of PPR. This might be due to the number of the goats (>1% of the flock) and keeping them separate from the sheep. The present article reviews the details of this outbreak in Iran.     

Clinical and Para‐clinical Findings of a Recent Outbreaks of Peste des Petits Ruminants in Iran

Peste des petits ruminants (PPR) is a highly contagious and infectious viral disease of domestic and wild small ruminants characterized by fever, erosive stomatitis, conjunctivitis, gastroenteritis and pneumonia. Goats are usually more severely affected than sheep. Peste des petits ruminants is caused by a paramyxovirus of the Morbillivirus genus. In March 2004, a flock of sheep in Tehran province with 430 deaths was visited. According to the history taken from the owner, at disease onset most of the deaths were recorded from adult sheep, 3 weeks later lambs (2 weeks to 4 months of age) showed the highest death rate. All animals from 3 months age received rinderpest vaccine 1 month after onset. Many of the lambs died just a few hours after their first sucking of the colostrum from infected mothers. Most of them showed very acute form of disease and died a few hours after onset of clinical signs. In clinical examinations most of the cases showed severe depression, high fever (41°C), anorexia, mocopulurent nasal discharge, erosive and necrotic stomatitis (dental path, hard palate and cheeks), diarrhoea and dehydration. Para‐clinical findings including histopathological, serological and haematological examinations also confirmed the presence of PPR in this flock. PPR outbreaks have been frequent in Iran in recent years. Further, we suggest that PPR is not a recent invader of Iran. The main difference in clinical signs between this outbreak and the same in other reports is that goats did not show any obvious signs of PPR. This might be due to the number of the goats (>1% of the flock) and keeping them separate from the sheep. The present article reviews the details of this outbreak in Iran.     

Experimental studies of chronic pneumonia of sheep

Strains of Mycoplasma ovipneumoniae and Pasteurella haemolytica isolated from sheep affected with chronic pneumonia were inoculated by endobronchial route to conventionally-reared and SPF (Specific Pathogen-Free) lambs. Changes resembling those of the naturally-occurring disease were produced in most lambs given the organisms in combination and in some given M. ovipneumoniae alone. Similar but less extensive changes were seen in SPF lambs and fewer animals were affected. Different strains of M. ovipneumoniae did not affect the extent of changes produced in SPF lambs. M. ovipneumoniae became established in the lungs of both types of sheep; P. haemolytica did so less readily.

It was concluded that chronic pneumonia may be reproduced in conventional animals by combined inoculation of M. ovipneumoniae and P. haemolytica. Age and status of immunity to mycoplasmas may account for the different responses of conventional and SPF lambs.     

Inherited chondrodysplasia in Texel sheep

CASE HISTORY: A skeletal disease characterised by dwarfism, limb deformity and sometimes sudden death occurred over a period of 5 years in lambs born on a commercial sheep farm in Southland. The disease showed variable expression and occurred in crossbred sheep. A genetic aetiology was supported by the birth of affected lambs over two seasons in a flock of putative carrier and affected sheep transported to Massey University. CLINICAL FINDINGS: Affected lambs appeared normal at birth but showed evidence of dwarfism, wide-based stance and exercise intolerance as early as 1 week of age. Most died within the first 3 months of life, often after developing bilateral varus deformity of the forelimbs. Some severely-affected lambs died suddenly of respiratory embarrassment, probably due to tracheal collapse. Mildly-affected individuals had a short, blocky stature and some survived to breeding age. PATHOLOGICAL FINDINGS: Gross and microscopic lesions of variable severity were present in the tracheal, articular, epiphyseal and physeal cartilages. In severe cases, articular cartilage in major joints was eroded from weight-bearing surfaces. The trachea was flaccid, abnormally kinked, and had thickened cartilaginous rings and a narrow lumen. Affected sheep that survived to breeding age eventually developed severe degenerative joint disease. Histologically, chondrocytes were disorganised, surrounded by concentric rings of abnormal fibrillar material, and the matrix often contained focal to coalescing areas of chondrolysis. DIAGNOSIS: Inherited chondrodysplasia of Texel sheep. CLINICAL RELEVANCE AND CONCLUSIONS: This chondrodysplasia differs from those previously described in sheep and is considered to be a newly-recognised, recessively-inherited genetic disease of the Texel breed. A defect in the synthesis of glycosaminoglycans in cartilage matrix is suspected. This disease of sheep may provide a suitable model for studying various forms of therapy for human chondrodysplasias.     

Inherited chondrodysplasia in Texel sheep

CASE HISTORY: A skeletal disease characterised by dwarfism, limb deformity and sometimes sudden death occurred over a period of 5 years in lambs born on a commercial sheep farm in Southland. The disease showed variable expression and occurred in crossbred sheep. A genetic aetiology was supported by the birth of affected lambs over two seasons in a flock of putative carrier and affected sheep transported to Massey University.

CLINICAL FINDINGS: Affected lambs appeared normal at birth but showed evidence of dwarfism, wide-based stance and exercise intolerance as early as 1 week of age. Most died within the first 3 months of life, often after developing bilateral varus deformity of the forelimbs. Some severely-affected lambs died suddenly of respiratory embarrassment, probably due to tracheal collapse. Mildly-affected individuals had a short, blocky stature and some survived to breeding age.

PATHOLOGICAL FINDINGS: Gross and microscopic lesions of variable severity were present in the tracheal, articular, epiphyseal and physeal cartilages. In severe cases, articular cartilage in major joints was eroded from weight-bearing surfaces. The trachea was flaccid, abnormally kinked, and had thickened cartilaginous rings and a narrow lumen. Affected sheep that survived to breeding age eventually developed severe degenerative joint disease. Histologically, chondrocytes were disorganised, surrounded by concentric rings of abnormal fibrillar material, and the matrix often contained focal to coalescing areas of chondrolysis.

DIAGNOSIS: Inherited chondrodysplasia of Texel sheep.

CLINICAL RELEVANCE AND CONCLUSIONS: This chondrodysplasia differs from those previously described in sheep and is considered to be a newly-recognised, recessively-inherited genetic disease of the Texel breed. A defect in the synthesis of glycosaminoglycans in cartilage matrix is suspected. This disease of sheep may provide a suitable model for studying various forms of therapy for human chondrodysplasias.     

Properties of a respiratory syncytial virus isolated from a sheep with rhinitis

A virus isolated from a yearling cross-bred ewe was identified as respiratory syncytial virus (RSV) by indirect immunofluorescence and by virus neutralization with bovine RSV antisera. The virus caused a mild conjunctivitis in 3-month-old lambs when inoculated alone. Although clinical signs of pneumonia were not observed, there was gross and microscopic evidence of pulmonary inflammation in the lungs of lambs inoculated with either the sheep RSV isolate alone or in conjunction with Pasteurella haemolytica. Lung lesions in the dual infection were more severe, with approximately 10% of the total lung mass affected. Lavage fluids from lambs inoculated with virus and bacteria contained approximately 3 times more inflammatory cells than from control lambs or lambs inoculated with virus only. The sheep RSV isolate was classified as a mild respiratory pathogen in lambs of this age. Speculations on the potential importance of this virus in interspecies transmission to cattle and goats were discussed.     

An outbreak of rickets in Corriedale sheep: evidence for a genetic aetiology

CASE HISTORY: A skeletal disease characterised by lameness, limb deformities and reduced growth rate occurred over two successive years in lambs born on a commercial sheep farm in Marlborough. A genetic aetiology was considered likely following exclusion of other known causes of rickets and because of the progressive nature of the disease, even after affected animals were transferred to another property. CLINICAL FINDINGS: Affected lambs appeared normal at birth but developed clinical signs during the first 2 months of life. The most severely affected animals either died or were euthanised within the first year of life, but some survived to breeding age. Serum biochemistry revealed hypocalcaemia, hypophosphataemia and increased concentrations of 1,25 dihydroxyvitamin D. The mean serum 25 hydroxyvitamin D concentration was similar to that of control lambs. PATHOLOGICAL FINDINGS: Gross lesions included enlarged costochondral junctions, bilateral irregularity of articular surfaces on humeral heads due to collapse of subchondral bone, thickened cortices in long bones and irregular thickening of physeal cartilages. Microscopically, tongues of hypertrophic chondrocytes extended from physes into metaphyseal regions; metaphyseal trabeculae were thick, disorganised and often lined by wide osteoid seams. Osteoclastic activity was excessive both in cortical and trabecular bone. DIAGNOSIS: Inherited rickets in Corriedale sheep. CLINICAL RELEVANCE AND CONCLUSIONS: This disease is likely to be present in several Corriedale sheep flocks in New Zealand and may have been misdiagnosed as arthritis or other diseases causing lameness and/or poor growth. A defect in end-organ responsiveness to 1,25 dihydroxyvitamin D is the likely mechanism. This disease of sheep may be a useful model for studying vitamin D metabolism and the treatment of inherited forms of rickets in human beings.     

Tick-Borne Fever in Lambs of Different Ages

Four groups of lambs aged 1 week, 4 weeks, 1/2 year and 1 year old respectively were inoculated with Ehrlichia phagocytophila infected blood. Clinical signs, temperature reactions, haematological changes and parasitaemia were more moderate in lambs inoculated with E. phagocytophila at the age of 1 week than those recorded in the older animals. The clinical response to tick-borne fever (TBF) appears to be more severe with increasing age of the lambs. The lymphocyte reactivity to mitogens was reduced in the TBF infected lambs, and was most pronounced in lambs in the 3 older age groups.     

Experimental Visna in Icelandic lambs.

Icelandic lambs were inoculated intracerebrally with visna virus at 10 days of age and sacrificed 22–25 days after infection. Virus was isolated from all 5 inoculated animals, and pronounced periventricular lesions, typical of early visna, were seen in the 3 lambs with the most widespread infection. The response to the infection was very similar to that seen previously in adult Icelandic sheep.In addition, 2 uninoculated lambs housed with the injected animals and sacrificed at the same time also became infected.     

First isolation of Mycoplasma capricolum subsp. capricolum, one of the causal agents of caprine contagious agalactia, on the island of Lanzarote (Spain)

During an unusually long period of bad weather, several outbreaks of caprine contagious agalactia (CCA) were reported in a number of flocks on the island of Lanzarote (Canary Islands, Spain). Clinical and subclinical mastitis in lactating goats and some cases of arthritis and pneumonia in kids were observed in the affected flocks. Mycoplasma capricolum subsp. capricolum was isolated as the main causal agent of the outbreaks, associated with M. mycoides subsp. mycoides "large colony type" (Mmm LC) in two flocks. This is the first report of an isolation of M. capricolum subsp. capricolum on the island of Lanzarote. The finding is of epidemiological importance and could complicate plans to control the disease. The significance of this mycoplasma species in association with CCA must now be studied in detail.     

Observations on the alimentary tract of gnotobiotic lambs.

Seventeen gnotobiotic lambs were reared up to 21 weeks of age on cows' milk followed by sterile solid diets similar to diets fed to conventional lambs. Seven were inoculated with limited defined populations of rumen bacteria, seven were left uninoculated and three were dosed with rumen contents from conventional sheep ('conventionalised'). Seven naturally-born lambs were reared for purposes of comparison. As with other species of gnotobiotic animals, both the inoculated and the uninoculated gnotobiotic lambs had small, poorly developed lymph nodes, soft colon contents and thin intestinal walls. Unlike other species the caeca of gnotobiotic lambs were of normal size. The overall size of the reticulo-rumen including contents relative to body weight was similar in gnotobiotic and conventional lambs. However, macroscopically, the musculature of the rumen seemed to be poorly developed and histological studies showed hypoplasia of the muscle tissue of both the rumen and reticulum. Rumination was noted only infrequently in gnotobiotic lambs. The epithelium of the rumen and reticulum of the uninoculated gnotobiotic lambs was similar to that of neonatal lambs, but there was normal development of papillae in gnotobiotic lambs inoculated with limited defined populations of rumen bacteria and in conventionalised lambs. Degenerative changes were observed histologically in some of the organs of gnotobiotic lambs which were consistent with nutritional deficiencies.     

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.     

Infection of specific-pathogen-free lambs with parainfluenza virus type 3, Pasteurella haemolytica and Mycoplasma ovipneumoniae

Groups of specific-pathogen-free lambs were inoculated with combinations of parainfluenza virus type 3 (PI₃), Pasteurella haemolytica (P.h.) and Mycoplasma ovipneumoniae ( (M.o.). Acute, necrotising bronchopneumonia developed in 8/9 lambs inoculated with PI₃ followed by P.h. whereas only 1/5 lambs inoculated with PI₃ followed by a combination of M.o. and P.h. developed a pneumonic lesion. When M.o. was inoculated 29 days before PI₃ and P.h., pneumonia developed in 3/4 lambs but M.o. was not reisolated from any of the lungs. Pneumonia was observed in 1/5 lambs inoculated with P.h. alone and in 1/5 inoculated with M.o. plus P.h. In addition, one lamb in the latter group died of acute septicaemic pasteurellosis. None of the lambs inoculated with M.o. alone, PI₃ alone or PI₃ followed by M.o. had any gross or microscopic evidence of pneumonia although the virus alone, or in combination, did produce minor pulmonary lesions.These data suggest that M.o. is not an important primary or secondary lung pathogen.     

Studies on the pathogenicity of bovine herpesvirus type 5 in sheep.

Four Merino lambs were intranasally inoculated with bovine herpesvirus type 5 (BHV-5) reference strain N569. Two lambs were mock-inoculated as negative controls. The virus-inoculated animals developed apathy, inappetence, rhinitis, nasal, ocular and genital discharge, slight diarrhea and neurological disorders, like tremor and salivation. BHV-5 was isolated from the nasal discharge in two of the animals, while the polymerase chain reaction (PCR) detected the virus in all the infected lambs. Two lambs died on post infection day (PID) 13, while the other two infected animals were euthanized on PID 15 and 30. Gross pathological changes were not observed, however, histopathological examinations revealed diffuse nonsuppurative meningo-encephalitis in all infected animals. Viral antigen was detected by immunohistochemistry and viral nucleic acid was revealed by in situ hybridization in the brain of the two lambs, which died on PID 13. The virus was demonstrated by virus isolation and by PCR from different organs of all the infected animals. Slight rise of antibodies was observed in the infected animals from PID 15. The results show that BHV-5 is able to cross the species barrier and may establish infection in sheep.     

Post natal ontogeny of immunological responsiveness in Merino sheep

Sheep in five age groups (two weeks, 10 weeks, 18 weeks, six months and four years old) were immunised systemically, twice, with ovalbumin or Brucella abortus (live or killed) and antibody responses in blood were measured. The animals were also infected with the nematode parasites Trichostrongylus colubriformis and Haemonchus contortus and faecal egg counts and serum antibody responses to larval antigens were measured. The experiments were designed so that, as far as possible, the effect of age per se could be dissociated from the combined effects of age and prior exposure to antigen. The effects of the age of sheep were more marked for antibody responses to Brucella abortus lipopolysaccharide than to ovalbumin. Older animals had much greater resistance to infection with internal parasites, as shown by the magnitude of the faecal egg count. In contrast to older lambs, neonatal lambs (infected with H contortus at two weeks old) had consistently declining concentrations of anti-H contortus antibody in their serum, mounted no detectable autogenous anti-H contortus antibody response in blood and appeared to develop no resistance to the parasite. Post natal ontogeny of immune responses was different for the various antigens/pathogens.     

Persistence of Ehrlichia phagocytophila infection in two age groups of lambs

Tick-borne fever (TBF) is caused by the rickettsiae Ehrlichia phagocytophila and is a common disease in sheep in tick (Ixodes ricinus) infested areas in Norway. Earlier investigations have shown that some sheep could remain infected for several months after the primary infection. In this study, the persistence of E. phagocytophila after experimental infection was investigated in 2 age groups of lambs. Six lambs (1-2 weeks old) and 14 lambs (6-8 months old) were inoculated intravenously with an ovine strain of E. phagocytophila and thereafter examined clinically (including daily body temperature recording) and by haematological and serological (E. equi antibodies) methods for the next 4 months. At the end of this period, the lambs were examined for a TBF infection by blood smear investigation and blood inoculation studies. The infection was demonstrated in 19 (95%) of the 20 lambs.     

Transmission of ovine herpesvirus 2 among adult sheep

Previous studies from this laboratory have defined the pattern of acquisition of ovine herpesvirus 2 (OHV-2) in lambs under natural flock conditions. This study examined the question of whether OHV-2 could be transmitted between adult sheep. Two potential routes of transmission were examined: (1) direct inoculation of either viable leukocytes or whole blood from OHV-2 positive sheep, and (2) horizontal transmission through natural contact with OHV-2 positive sheep. Two groups of OHV-2 negative adult sheep were inoculated with material from infected sheep, one with 5x10(8) viable peripheral blood leukocytes (PBL), and the other with 100 ml of whole peripheral blood. No PCR signals were detected in any of the three sheep inoculated with the PBL during the 20 weeks following inoculation. In the group of five sheep inoculated with whole blood, two became PCR-positive at 7 and 8 weeks post-inoculation, respectively, and the remaining three sheep maintained their negative status until termination of the experiment at 20 weeks post-inoculation. In two experiments conducted in different flocks, a total of 20 adult sheep were used to examine horizontal transmission by contact; all animals became PCR-positive within 12 months of mixing the uninfected and infected animals. The results of these experiments support two conclusions. First, the susceptibility to OHV-2 is not limited to young lambs; adult sheep remain fully susceptible. Second, the fact that whole blood, but not PBL, from infected sheep was able to transmit the infection to only two of five inoculated sheep suggests that the infection in peripheral blood cells may be largely non-productive.