The isolation, propagation and characterization of tissue-cultured equine rotaviruses

From 105 field cases of diarrhea in neonatal or young foals, rotavirus was detected by electron microscopy (EM) and/or by enzyme-linked immunosorbent assay (ELISA) in the feces of 65 foals on 16 different premises. ELISA was performed with Rotazyme test kits developed by Abbot and Company for the detection of rotaviruses. Twenty-four field isolates from the feces of diarrheic foals with equine rotavirus infection as ascertained by EM were placed in MA-104 cell cultures after pretreatment of the viral suspension with 10 micrograms ml-1 of trypsin and incorporation of 0.5 micrograms ml-1 or 1 microgram ml-1 of trypsin in Earle's minimal essential medium (MEM), 2% lactalbumen hydrolysate, and antibiotics. The isolates that replicated in cell culture produced varying degrees of cytopathic effect. After the 24 isolates had been transferred 5 or 7 times in cell culture, viral particles were observed in 17 by EM, and 22 had positive ELISA tests as determined by visual color chart and spectrophotometric readings. Concentrated tissue-cultured viral antigen of 9 isolates fixed complement using Nebraska calf diarrhea rotavirus calf antiserum while four isolates gave negative results. The same 13 tissue-cultured viral suspensions failed to fix complement using reovirus antiserum. The 9th passages of two isolates (EID1 and EID2) yielded titers of 10(4.45) ml-1 TCID50 and of 10(4.95) ml-1 TCID50, respectively, as measured by cytopathic effect. After 13 tissue-cultured passages, 2 other isolates, EID3 and EID4, each had titers of 10(6.2) ml-1 TCID50 and of 10(5.95) ml-1 TCID, respectively. Cytoplasmic or intranuclear inclusions were not seen in any cells of the MA-104 infected cell cultures. Small, but distinct, plaques in MA-104 cell cultures were produced by the EID1 isolate. Polyacrylamide gel electrophoresis tests of EID1 and EID2 isolates at the 9th cell passage and EID3 and EID4 isolates at the 13th cell passage each showed that the RNA genome had 11 segments with a migrating pattern that was identical for each isolate and characteristic of rotaviruses. These 4 equine tissue-cultured isolates when tested by ELISA, utilizing a monoclonal antibody serum pool that cross-reacted with many rotavirus isolates, each gave positive values comparable to rotavirus antigen controls.     

An unusual case of inclusion body hepatitis in a cockerel

This report describes inclusion body hepatitis in a 12-week-old leghorn-type cockerel. The cockerel had come from a specific-pathogen-free flock and was reared in a positive-pressure isolator until 6 weeks old. Grossly, the cockerel was jaundiced, and the liver was swollen and had small white foci throughout. Histologically, there were foci of hepatic necrosis and basophilic inclusion bodies in the nuclei of numerous hepatocytes. A fowl adenovirus, isolated from the liver, was found to belong to serotype 2/12. The bursa of Fabricius had some degenerative changes, but these were not consistent with infectious bursal disease, nor was there evidence of seroconversion to infectious bursal disease virus in the remaining chickens.     

Herpes viral hepatitis in a toucan

Herpesvirus infection was diagnosed in a toucan. The herpesvirus was isolated from the liver and identified by electron microscopy in the liver and in cell culture. A negative immunofluorescent reaction was obtained when virus-infected cell cultures were reacted with a conjugate to the herpesvirus of Pacheco's disease. The main pathologic finding in the toucan consisted of a severe necrotizing hepatitis with intranuclear inclusions in the liver and spleen. A presumptive diagnosis of chlamydiosis was also made, based on a positive direct fluorescent monoclonal antibody reaction to chlamydial antigens in impression smears of liver and spleen. Chlamydial isolation attempts were unsuccessful. The toucan had been in contact with two macaws that had died 5 days before the toucan died and were diagnosed by histology as having herpesvirus hepatitis.     

Cataracts and optic nerve hypoplasia in turkey poults

Newly hatched commercial turkey poults culled because of grossly visible cataracts were studied. A total of 43 affected and 23 unaffected control poults at various ages were necropsied, and the ocular changes in affected poults were compared with those of aged-matched controls. Affected poults had consistent cataracts coupled with a marked depletion in retinal inner plexiform, ganglion cell, and optic nerve fiber layers, with a resultant reduction in the size of the optic nerves. Lesions were seen in 1-day-old poults. Lens changes included microphakia and lens fiber degeneration throughout the lens, with nuclear liquefaction. The depletion in the numbers of retinal ganglion cells did not appear to progress over several weeks time. The ganglion cell depletion was not uniform within the retina. The cause for these ocular changes is unknown.     

Assessment of ram fertility in hill flocks in Argyll

Examination of 1260 rams, involving 1540 semen examinations, revealed that 9 per cent of rams in hill flocks in Argyll were subfertile as assessed by semen evaluation. Although 34 lesions of the genitalia were identified, only 13 were associated with poor semen quality. A five year survey of one blackface flock resulted in the elimination, after three years, of a 17 per cent level of infertility following routine examination of all rams, with an increase of 2.27 per cent in the lamb crop. Based on the 1981 lamb prices for this flock an increased income of 1085 pounds was achieved.     

Evaluation of two test procedures for diagnosis of giardiasis in dogs.

Two commercially available tests, an antigen-capture ELISA for use on fecal samples, and a peroral nylon string test for use in dogs, were compared with a zinc sulfate fecal concentration technique (ZSCT) for detection of giardiasis in dogs. Of 77 dogs and 164 fecal samples (from these dogs), 33 and 52, respectively were found to be Giardia-positive on the basis of results of the ZSCT. The ELISA gave false-negative results for 10 and 14% of ZSCT-positive dogs and fecal samples, respectively, and false-positive results (relative to the ZSCT test results) in 13 and 10% of ZSCT-positive dogs and fecal samples, respectively. Of the 18 string-tested dogs, 14 were positive by results of the ZSCT. Of the 4 dogs that were Giardia-negative by ZSCT, 2 were Giardia-positive by ELISA. Dogs were sedated and given water and metoclopramide to aid passage into the duodenum of the capsule containing a nylon string. Of the 21 string tests performed on the 18 dogs, only 5 strings reached the duodenum, and 0 of the 5 yielded positive results for Giardia sp. Because the string broke in 1 dog (leaving most in the gastrointestinal tract and, therefore, producing a risk of string foreign body) further string tests were not done.     

Long-term survival after combination chemotherapy for bilateral renal malignant lymphoma in a dog

CASE HISTORY: A 6-year-old, entire male Flat-coated Retriever was presented with a history of lethargy, polydipsia and seizures. Clinical chemistry had shown marked azotaemia.

CLINICAL FINDINGS AND DIAGNOSIS: Radiography and ultrasonography revealed bilateral renomegaly, and cytology of fine needle aspirates from the kidneys was diagnostic of malignant lymphoma. The dog was treated with a modified high-dose cyclophosphamide-, vincristine-, and prednisolone-based chemotherapy protocol, achieved remission, and returned to normal quality of life. Survival time was 346 days from the time of diagnosis.

CLINICAL RELEVANCE: Malignant lymphoma in the kidneys of dogs has been considered to carry a uniformly poor prognosis. Long-term remission after medical treatment has not previously been reported. The favourable outcome in this case illustrates the limitations of clinical staging in determining the outcome for individual patients.