Effects of cyclosporine A on reovirus-infected broilers

The effect of cyclosporine A on reovirus-infected male broiler chickens was studied. Beginning at 1 or 10 days of age, 3 groups of 15 broilers were injected in the pectoral muscle with 50 mg of cyclosporine A (CSA) in oil per kg body weight every 3 days until 28 days. Controls were injected with olive oil. Two CSA-injected groups and one untreated group were orally infected with 1000 TCID50 of reovirus at 1 day of age. Cell-mediated immunity was evaluated at 17 and 24 days by a delayed-wattle-response test to injected phytohemagglutinin (PHA-M). Cyclosporine A and reovirus significantly (P less than 0.001) depressed the wattle response following the first injection of PHA-M but not the second. At necropsy 28 days postinoculation (PI), no gross lesions were apparent. Histologic lesions in birds infected with reovirus were lymphocytic pericarditis and tenosynovitis; synovial cells were hyperplastic, and heterophils and fibrin were in synovial spaces. Thymic medullary diameters were significantly (P less than 0.001) smaller in all CSA-treated birds. Although CSA suppressed cell-mediated immunity somewhat, there were no apparent differences in severity of microscopic lesions among reovirus-infected groups.     

In vitro and in vivo characterization of avian reoviruses. III. Host factors affecting virulence and persistence

Three avian reovirus isolates (2177, 2035, and 1733) were used to determine the effect of the age of chickens at inoculation on virus virulence and persistence. Groups of specific-pathogen-free leghorns were inoculated with three different reovirus isolates of different levels of pathogenicity at 1 day, 1 week, 2 weeks, 3 weeks, or 4 weeks of age. Tissues were examined for the presence of virus and lesions at regular intervals until 8 weeks postinoculation (PI) and then again at 22 weeks PI. Isolate 1733, which is highly pathogenic, was reisolated from the thymus, trachea, liver, intestine, cecal tonsils, bursa of Fabricius, gastrocnemius tendon, and white blood cells. Microscopic lesions were observed in some tissues, including the thymus, liver, spleen, bursa of Fabricius, and gastrocnemius tendons, when sampled within a 7-day period following inoculation. This isolate persisted and produced microscopic lesions in the gastrocnemius tendons for as long as 22 weeks PI. The isolates of intermediate pathogenicity (2035) or low pathogenicity (2177) were isolated less frequently and from fewer tissues than isolate 1733. Isolate 2035 could be found in the gastrocnemius tendons as long as 7 weeks PI, whereas isolate 2177 was never isolated from the tendons, nor did it produce any notable gross or microscopic tissue changes. Birds inoculated at age 1 week or older with any of the three reovirus pathotypes were more resistant to infection than 1-day-old inoculates, as evidenced by a decrease in virus reisolations and a concurrent reduction in the severity of lesions in selected tissues.     

Infectious tenosynovitis in broilers and broiler breeders in Egypt

Two infectious tenosynovitis-producing viruses were isolated from tendon sheaths and synovial fluids of 59 broilers and 15 broiler breeders obtained from different flocks in Egypt during June to October 1983. The viruses grew well on the chorioallantoic membrane of developing chicken embryos, produced small localized white pock lesions with oedematous swellings at the inoculation sites and death of most of the embryos 72 to 96 hours post-inoculation. They also induced cytopathic effect in chicken embryo rough, Vero and MS cell lines. The viruses were neutralized by reovirus S1133 antiserum, both in tissue culture and. on the chorioallantoic membrane. Inoculation of the viruses into 2-day-old broiler chicks via the foot pad, intramuscular and oral routes reproduced the disease with the development of characteristic clinical, pathological and serological responses. The infection was transmitted to in-contact control chicks. This is the first report of the disease and of the isolation and identification of the causative virus in Egypt.     

Infectious tenosynovitis in broilers and broiler breeders in Egypt

Two infectious tenosynovitis-producing viruses were isolated from tendon sheaths and synovial fluids of 59 broilers and 15 broiler breeders obtained from different flocks in Egypt during June to October 1983. The viruses grew well on the chorioallantoic membrane of developing chicken embryos, produced small localized white pock lesions with oedematous swellings at the inoculation sites and death of most of the embryos 72 to 96 hours post-inoculation. They also induced cytopathic effect in chicken embryo rough, Vero and MS cell lines. The viruses were neutralized by reovirus S1133 antiserum, both in tissue culture and on the chorioallantoic membrane. Inoculation of the viruses into 2-day-old broiler chicks via the foot pad, intramuscular and oral routes reproduced the disease with the development of characteristic clinical, pathological and serological responses. The infection was transmitted to in-contact control chicks. This is the first report of the disease and of the isolation and identification of the causative virus in Eqypt.     

Increased cartilage oligomeric matrix protein concentrations in equine digital flexor tendon sheath synovial fluid predicts intrathecal tendon damage

Objectives: To evaluate digital flexor tendon sheath (DFTS) synovial fluid cartilage oligomeric matrix protein (COMP) concentrations as a molecular marker for intrathecal pathology. Study Design: Case control study. Animals: Horses (n=46) with DFTS tenosynovitis; 23 fresh cadaver horses. Methods: DFTS synovial fluid samples were collected from clinical cases with noninfected DFTS tenosynovitis and from control DFTS. Clinical and surgical findings were recorded, and dissection of control limbs was performed to confirm the DFTS to be grossly normal. Synovial fluid COMP was quantified using a homologous competitive inhibition ELISA. Results: Abnormalities were identified tenoscopically: intrathecal tendon/ligament tearing was identified in 37 cases and 9 had other lesions. In control horses, synovial fluid COMP was higher in younger horses. Clinical cases with intrathecal tendon/ligament tearing had higher synovial fluid COMP than either clinical cases with other lesions, or controls. In horses ≥5 years old, the sensitivity and specificity of the assay was high for diagnosing intrathecal tendon/ligament tearing. Conclusions: COMP concentrations in DFTS synovial fluid were significantly greater than those in normal horses with noninfected tenosynovitis caused by intrathecal tendon/ligament tearing, but not by other lesions.     

Enteropathogenicity of Dutch and German avian reoviruses in SPF white leghorn chickens and broilers

The enteropathogenicity of avian reoviruses (ARVs), isolated from chickens affected with malabsorption syndrome (MAS) from The Netherlands and Germany was studied. In the first trial seven different ARVs isolated from MAS cases were inoculated in 1-day-old specific pathogenic free (SPF) white leghorns. The pathogenicity was compared with 2 ARVs isolated from cases of tenosynovitis, namely reference strain S1133 and a Dutch strain. Although a difference in the severity of the clinical disease was observed, all reoviruses could induce vacuolar degeneration and sloughing of the epithelium of the small intestine at day 2 post inoculation (PI) till day 7 PI. Two Dutch and one German ARV derived from MAS causing the most severe intestinal lesions at day 2 PI, were further studied in the second trial using SPF broilers. These reoviruses did not cause weight gain depression in the broilers although lesions in the small intestine were present from day 1 up to day 4 PI and were more severe than in the white leghorn chickens. In one of the inoculated groups apical denuded villi were already present at day 1 PI. At day 7 PI the small intestine of the infected broilers appeared to be normal. Reovirus antigen was detected in the cytoplasm of the enterocytes at the tip and middle section of the affected villi both in layers and in broilers. To study the role of intestinal CD4+ and CD8+ T-cells and macrophages/monocytes in the pathogenesis of ARV, the numbers of these cells of the jejunal villi of one infected and the control broiler groups were compared. CD4+ T-cells were detected in low numbers and only in the infected broiler group at day 14 PI. The numbers of CD8+ T-cells and macrophages/monocytes were significantly higher in the infected broiler group than in the control broiler group at day 7 and 14 PI and at day 7 PI respectively. Our study indicates that the reovirus alone cannot induce intestinal lesions as found in MAS chickens. Moreover, CD8+ T-cells may play a major role in the pathogenesis and or reovirus clearance in the small intestine.     

Interaction of reovirus and Cryptosporidium baileyi in experimentally infected chickens

No clinical signs, gross lesions, or increased mortality were observed in specific-pathogen-free chickens orally inoculated at 5 days of age with Cryptosporidium baileyi, reovirus 2035, reovirus 2408, or combinations of these agents. Weight gain of chickens inoculated with only reovirus 2408 was depressed 0-8 days postinoculation (PI) (P less than 0.01) but not for the 21-day period PI. Weight gain of chickens inoculated with only reovirus 2035 was not affected. Cryptosporidium baileyi infection significantly depressed weight gain 8-14 days PI but not for the entire 21-day period PI. Weight gain of chickens infected with both C. baileyi and reovirus 2035 was significantly depressed 0-14 days PI and for the entire 21-day period PI. Dual infection with C. baileyi and either reovirus appeared to promote shedding of both agents. Cryptosporidia were found principally in the rectum 2-10 days PI and in the bursa of Fabricius 6-10 days PI. Reovirus infection did not cause any microscopic lesions and did not modify lesions caused by C. baileyi infection.     

Experimental reovirus hepatitis in newborn chicks.

The avian reovirus "UM 1-203" originally isolated in the United States from chickens with tenosynovitis was pathogenic for the newborn chick infected by parenteral inoculation. It induced plurivisceral lesions, which became particularly intense in the liver. The intense local multiplication of the virus provoked a necrotizing hepatitis; viral titers were maintained around an E.I.D.50 of 10(8)/0.2 ml of organ suspension in chicks killed between the 3rd and 5th days after inoculation. As a specific cellular response to the viral multiplication, numerous polykaryocytes formed and increased in number, then regressed and disappeared from the hepatic parenchyma. By histologic and electron microscopic examination at progressive times after infection, the virus was recognizable in the polykaryocyte cytoplasm. In chicks that survived longer (killed the 5th and 6th days after inoculation), regeneration of the hepatic parenchyma occurred and seemed to develop from groups of dense hepatocytes that either lacked the virus or survived the acute phase of infection.     

Comparative study of the pathogenicity of avian reoviruses

Reovirus strains CO8 and 81-5, isolated from chickens with malabsorption syndrome, and reovirus strain 176, isolated from chickens with tenosynovitis, were each individually inoculated into 1-day-old specific-pathogen-free chicks. Strain CO8 caused tenosynovitis and pericarditis following footpad inoculation, but it was of low pathogenicity when given by either oral or subcutaneous route. In contrast, strains 81-5 and 176 were highly pathogenic and caused severe mortality following subcutaneous inoculation. Lesions included hepatic necrosis, bursal atrophy, thymitis, and splenic hyperplasia; strain 81-5 also caused pericarditis and myocarditis. Although strain 176 caused higher mortality than strain 81-5, these two strains differed little in the severity and distribution of the lesions they caused. No signs or lesions of typical malabsorption syndrome were observed.     

Influence of a 12.5 per cent rapeseed diet and an avian reovirus on the production of leg abnormalities in male broiler chickens

The incidence of different forms of leg abnormality were recorded in reovirus (S1133) infected and control male broiler chickens fed on a normal commercial diet or one of similar nutritive value containing 12.5 per cent rapeseed meal. Regular serological examination showed that birds remained free from Mycoplasma gallisepticum and M synoviae infection throughout the 10 week period of investigation. Precipitating antibodies to the reovirus were detected in 90 per cent of the infected birds between the third and 10th week after infection. Carotene levels in rapeseed fed groups showed no significant differences between reovirus infected and control birds or between birds with or without clinical signs of leg abnormality. The most frequent and severe leg abnormalities were present in the infected birds fed on the rapeseed diet, followed by those fed on the normal commercial diet. There was a highly significant difference (P less than 0.01) between the number of birds with leg abnormalities in each of these groups and their corresponding control groups. The lesions which were mainly responsible for these differences were tenosynovitis and enlarged hocks. Oral infection with reovirus did not appear to make the birds more susceptible to other types of leg abnormality, although the severest lesions of dyschondroplasia were seen in birds which had been exposed to the dual effects of reovirus and rapeseed diet.     

Prevalence of reoviruses in commercial chickens

The prevalence of reoviruses in commercial chickens with the runting/stunting syndrome, tenosynovitis, and normal chickens was investigated. Reoviruses were isolated from 3-week-old chickens affected with the runting/stunting syndrome and from older chickens with tenosynovitis; viruses were isolated from tissues with and without lesions. Reoviruses were also frequently isolated from rectal contents of normal 3-week-old chickens, and there was serological evidence of previous reovirus infection in all flocks of adult meat breeder chickens examined. The widespread occurrence of reoviruses in both normal and diseased chickens indicates that the isolation of reoviruses from tissues of chickens with lesions does not necessarily imply any aetiological relationship of reovirus with disease, even in the absence of other known causes. However, the occurrence of reovirus in normal chickens does not preclude an aetiological relationship with disease and further investigation of strain variation and possible virulence factors in avian reoviruses is required.     

Clinical and pathologic analyses of bicipital tenosynovitis in dogs

OBJECTIVE: To determine the clinical and pathologic findings in dogs with primary bicipital tenosynovitis. ANIMALS: 19 dogs with 20 shoulder joints treated surgically for bicipital tenosynovitis and 8 shoulder joints from 4 clinically normal dogs. PROCEDURE: Histologic abnormalities of tendon sheaths of the biceps brachii in affected dogs were determined by use of comparison with findings in clinically normal dogs. Specimens were graded for inflammation, fibrosis, villous hypertrophy, vascular prominence, and synovial cell proliferation. Histopathologic results were statistically evaluated for relationship with clinical findings and treatment before surgery. RESULTS: Synovial villous hypertrophy and vascular prominence were the most consistent histologic findings in 16 and 14 of 20 affected joints, respectively. Evidence of inflammation was lacking in 6 joints. Ten joints had inflammatory cell infiltration of the tendon sheath. Plasma cells and lymphocytes were the most common infiltrates; however, the type and amount of inflammatory cell infiltrate were variable. Fibrosis of the tendon sheath was seen in 8 joints, and synovial cell proliferation was seen in 11 joints. Other changes included accumulation of hemosiderin, focal calcification, osseous metaplasia, lysis of collagen, and fibrocartilaginous metaplasia. No significant relationship was detected between histopathologic findings and clinical findings or treatment before surgery. CONCLUSIONS AND CLINICAL RELEVANCE: Inflammation was more variable than hypothesized and may not be a consistent pathophysiologic feature of bicipital tenosynovitis. In some dogs, this disease may be the result of a degenerative process rather than an inflammatory process.     

The effect on newborn chicks of oral inoculation of reovirus isolated from chickens with tenosynovitis

Reovirus strain 176, isolated from chickens with tenosynovitis, was highly pathogenic following oral inoculation of 1-day-old specific-pathogen-free chicks. Disseminated lesions including hepatic necrosis, splenic lymphostromal cell hyperplasia, and bursal atrophy occurred on day 3 postinoculation (PI), followed by myocarditis on day 6 PI and by pericarditis and tenosynovitis on day 9 PI. Reovirus was isolated from the liver as early as day 1 PI, whereas significant neutralizing antibody was detected on day 13 PI. Mortality occurred from day 4 to day 7 PI, and the death of birds was associated with the severity of hepatic necrosis. The occurrence of tenosynovitis in virus-inoculated birds was subclinical.     

Reovirus-induced tenosynovitis: persistence of homologous challenge virus in broiler chicks after vaccination of parents

Broiler chicks from a parent flock previously vaccinated with a commercial tenosynovitis (viral arthritis) vaccine were challenged when one day old with a virulent form of the vaccinal reovirus strain. A group from unvaccinated parents was similarly challenged. At three weeks after infection it was found that, while maternal antibody reduced the incidence of lesions of tenosynovitis by about 50 per cent and also the amount of virus in the gut when compared with the group without maternal antibody, the rates of recovery of virus from the hock joints were very similar. The possible epidemiological importance of persistent virus in the joints of clinically protected chicks is discussed.     

Severity of tenosynovitis in reovirus-infected chickens fed various dietary levels of choline, folic acid, manganese, biotin, or niacin

Five experiments were conducted to determine the incidence and severity of tenosynovitis in tendons distal to the tarsal joint in 4-, 6-, and 8-week-old reovirus WVU 2937-infected chickens fed diets containing 20%, 100%, or 200% of the 1977 National Research Council nutrient requirements (NCR-77) of manganese, biotin, niacin, choline, or folic acid. Male chickens, but not female chickens, fed 20% or 100% or the NRC-77 level of folic acid had consistently higher lesion scores of tenosynovitis than male chickens fed the 200% NRC-77 level of folic acid. Increasing dietary manganese levels from 20% to 200% NRC-77 reduced the severity of tendon swelling in 6- and 8-week-old male chickens and appeared to increase the severity of tendon swelling in 8-week-old female chickens. Male chickens fed 20% of the NRC-77 required level of biotin had numerically more severe tenosynovitis at 6 weeks of age and significantly more severe tenosynovitis at 8 weeks of age than male chickens fed 100% and 200% of the NRC-77 level of biotin. Increasing dietary choline levels from 20% to 200% of the NCR-77 required level increased the severity of tenosynovitis numerically at 6 weeks of age and significantly at 8 weeks of age. When data were pooled across diet, male chickens had significantly more severe tenosynovitis than female chickens at 4, 6, and 8 weeks of age in the choline experiment and at 4 and 6 weeks of age in the folic acid experiment. Differences in the severity of tenosynovitis of individual treatment means (sex X diet X infection interaction means) occurred at only the 20% or 100% NRC-77 level of choline or folic acid. Male chickens also had more severe tenosynovitis than female chickens when fed 20% of the NRC-77 required level of manganese for 6 or 8 weeks.     

Pathogenicity for chickens of a reovirus isolated from turkeys

A viral agent that was isolated from livers of commercial turkey poults that died at approximately two weeks of age was characterized as a reovirus. Experimental infection of day-old chickens with this reovirus isolate resulted in the development of tenosynovitis, hepatitis, and myocarditis. In vitro neutralization of the turkey reovirus isolate by antiserum against chicken reovirus correlated with in vivo protection of maternally immune chickens from day-old oral challenge with the turkey reovirus isolate.     

Histopathological, radiographic, and arthrographic comparison of the biceps tendon in normal dogs and dogs with biceps tenosynovitis

In dogs surgically treated for biceps tenosynovitis, the most common histopathological findings were fibrosis and collagen degeneration (n=13), synovial villous or vascular hyperplasia (n=10), lymphocytic-plasmacytic infiltrates (n=10), cartilaginous metaplasia (n=8), and ischemic necrosis (n=5). Degree of histopathological changes was associated with degree (p equals 0.000), but not duration (p equals 0.543), of lameness. Furthermore, there was no association between histopathological changes and age or radiographic and arthrographic findings. Cartilage metaplasia was the only histopathological finding in both affected tendons (8/18) and normal control dogs (13/13). Age and size of the control dogs were not determined; however, since all these dogs were clinically normal, fibrocartilaginous metaplasia can be present as an incidental finding in the biceps tendon of origin in dogs.     

Isolation of viruses from outbreaks of suspected tenosynovitis (viral arthritis) in chickens

Specimens from the legs of chickens from 84 outbreaks of suspected tenosynovitis were examined for the presence of viruses by culture in chick embryo lung or liver cell monolayers. All samples were from broilers or broiler breeders, ranging in age from dead-in-shell embryos to 36 weeks old. Twenty-five outbreaks (29.8 per cent) yielded viruses of which 12 were reoviruses alone, 12 adenoviruses alone, and one, a mixture of both types of virus. Rupture of the gastrocnemius tendon was seen in 12 outbreaks and viruses were isolated from six of these: three were reoviruses and three were adenoviruses. Approximately half the affected flocks from which specimens were received were in the six to 14 week age range. With one exception, all the reovirus isolations were made from chickens of 11 weeks or under, while adenovirus isolations showed more scatter with regard to age.     

Ultrastructure of the gastrocnemius tendon and sheath from broilers infected with reovirus

Gastrocnemius tendon and sheath from male broilers infected with reovirus at 1 day of age were evaluated ultrastructurally at 1, 2, 3, 4, 5, 10, 15, and 20 weeks postinoculation (PI). Although virus particles were not identified, degenerative changes in sheath and tendon fibroblasts were characterized by cytoplasmic vacuolization, disruption of membranes, and loss of ribosomes from rough endoplasmic reticulum, mitochondrial degeneration, and cell disruption.     

Isolation of avian reovirus as a possible etiologic agent of osteoporosis ("brittle bone disease"; "femoral head necrosis") in broiler chickens

Avian reovirus was isolated from intestines of 3-to-7-day-old broiler chickens with enteritis from broiler houses where osteoporosis was a problem. The virus was purified in a cesium chloride gradient (buoyant density 1.37 gm/ml) and identified as a reovirus by electron microscopy. Specific-pathogen-free (SPF) chickens and commercial broiler chickens with anti-reovirus maternal antibodies inoculated at 1 day of age with the reovirus isolate developed lesions of femoral head fractures and/or osteoporosis; reovirus could be reisolated from the bone marrow and intestinal tracts of experimentally infected SPF birds. The reovirus isolate, although isolated from intestines, induced development fo tenosynovitis lesions in SPF and commercial broiler chickens.