Histologic, immunohistochemical, and electron microscopic features of a unique pulmonary tumor in cockatiels (Nymphicus hollandicus): six cases.

A unique form of pulmonary malignancy develops in cockatiels. This report describes the gross, histologic, electron microscopic, and immunohistochemical features of this tumor in 6 cockatiels. DNA in-situ hybridization for polyomavirus in the neoplasm was also performed. The tumor was comprised predominantly of compact sheets of anaplastic round to polygonal cells. All tumors had a high mitotic index, and had occasional large clear to slightly basophilic intranuclear inclusions that caused peripheral dispersal or complete masking of chromatin. Tumors were invasive but convincing metastases were not observed. Transmission electron microscopy identified intracytoplasmic intermediate filaments, desmosomes between cells, and intranuclear cytoplasmic invaginations corresponding to the intranuclear inclusions in light microscopic sections. Neoplastic cells stained positive for vimentin, lysozyme, and in 1 bird, pan cytokeratin. All 6 pulmonary neoplasms were negative for avian polyomavirus using the FN-19/FN-40 cocktail and the long VP-1 probe. We propose that these tumors may be poorly differentiated carcinomas of pulmonary or thymic origin.

Effects of allopurinol on plasma uric acid levels in normouricaemic and hyperuricaemic green iguanas (Iguana iguana).

A two-phase cross-over therapeutic study was performed with 19 green iguanas (Iguana iguana) maintained within a preferred optimum temperature range of 26 to 37 degrees C. During phase 1, they were fed a normal vegetarian diet and medicated orally with either allopurinol or a placebo control once a day for seven days. Uric acid concentrations, total protein, packed-cell volumes (pcv) and bodyweights were recorded from each lizard before and after treatment to determine the effects of allopurinol. In phase 2, after a 10-day washout period, the iguanas were fed a high protein diet to induce hyperuricaemia. Normo- and hyperuricaemic iguanas that received 24.2 (3.2) mg/kg allopurinol had significantly lower mean (sd) uric acid concentrations (100.3 [53.1] micromol/l) than the controls (159.3 [100.3] micromol/l). There were no detectable interactions between the doses of allopurinol or placebo, and the iguanas' diet, weight, pcv or total protein. The allopurinol was well tolerated, and there was no significant clinical, gross or histological evidence of hepatic or renal toxicity in the iguanas that received the drug. However, in the kidneys of the hyperuricaemic iguanas that did not receive allopurinol there were proliferative changes in the glomeruli and degeneration of tubular epithelia. Allopurinol given orally at 25 mg/kg daily is able to reduce plasma uric acid levels by 41 to 45 per cent, and is therefore recommended for the treatment of hyperuricaemia in the green iguana.

Aleutian disease in two domestic striped skunks (Mephitis mephitis).

This report describes the use of polymerase chain reaction and DNA in situ hybridization to diagnose Aleutian mink disease parvovirus DNA in various tissue specimens from 2 companion striped skunks. Clinical, laboratory, and microscopic findings also support a clinical diagnosis of Aleutian disease in these mink.

Variation among pathologists in the histologic grading of canine cutaneous mast cell tumors with uniform use of a single grading reference.

Ten veterinary pathologists independently assigned histologic grades to the same 60 canine cutaneous mast cell tumors using the Patnaik classifications. The degree of agreement in grading among the pathologists was compared with the degree of agreement among the same pathologists in a previous study, in which each pathologist used the reference for grading that he/she uses routinely. Mean agreement improved significantly from 50.3% to 62.1% with uniform use of the Patnaik classifications (P = 0.00001), suggesting that there is value in uniform application of a single grading scheme for canine cutaneous mast cell tumors. Agreement among pathologists was still not 100%, suggesting that a more objective grading scheme should be developed and that other histologic indicators of prognosis should be investigated.

Pathology and histology of dietary tryptophan deficiency in broiler chicks.

The purpose of this experiment was to characterize a lesion of the rhamphotheca associated with tryptophan (TIRP) deficiency, search for other histological abnormalities,and determine whether bird size and housing conditions are contributing factors to these lesions. Day-old broiler chicks (Ross X Ross 308) were placed in either floor pens with fresh pine shavings or Petersime battery brooders with two pens of 10 chicks each per treatment. Broiler chicks from 0 to 21 days of age were fed adequate (0.24%) and deficient (0.09%) levels of TRP in diets based on corn, corn gluten meal, and gelatin. Separate groups of control chicks were pair fed daily with the deficient chicks. Deficient chicks grew less efficiently than did the pair-fed controls. Upon gross examination, a lesion of the maxillary rhamphotheca in the vicinity of the nares was observed in 61% of TRP-deficient birds housed in the battery and 13% of the birds housed in floor pens. A similar gross lesion was only observed in one control bird. These lesions were located along the upper portion of the beak between the nares and appeared as a crusty or scab-like area on gross examination, composed of detritus, heterophils, and plasma protein. Inflammation occasionally was observed at the dermoepidermal junction. The incidence of lesions was reduced in floor pens compared to battery brooders, but similarly sized birds did not exhibit the lesion. The number of lesions seen grossly and histologically in TRP-deficient birds, as compared to control birds, supports the hypothesis that TRP deficiency is the primary cause of these lesions around the nares of broilers. Secondary environmental factors, perhaps coprophagy, also influence the incidence of the lesion.

Hepatic hyaline globules in an Eclectus parrot (Eclectus roratus).

Hepatic hyaline globules, similar to those reported in some human livers, were observed in liver tissue from an Eclectus parrot (Eclectus roratus). The cytoplasmic inclusions were periodic acid-Schiff positive and diastase resistant and failed to stain by acid-fast or Congo red techniques. Ultrastructurally, the hepatic globules were composed of granular amorphous material with small peripheral striations that extended into the cytoplasm.

Tissue distribution and genetic typing of porcine circoviruses in pigs with naturally occurring congenital tremors.

Congenital tremors (CT) type A2 is associated with porcine circovirus (PCV) and deficient and abnormal myelin. The aim of this study was to determine the tissue distribution and genetic type of PCV in 1-2-day-old pigs with naturally occurring CT type A2 using in situ hybridization, polymerase chain reaction (PCR), and indirect fluorescent antibody tests on frozen tissue sections. CT-affected and clinically normal pigs were selected from 4 farms in the midwestern USA that were undergoing outbreaks of CT type A2. All CT and most normal pigs were infected with PCV. PCV was widely distributed in tissues of infected pigs and was most common in tissues of the central nervous system and liver. In all infected pigs, there were more PCV-infected cells in brain and spinal cord than in nonneural tissues. CT pigs had many more PCV-infected cells in the brain and spinal cord than did clinically normal pigs because of a more diffuse distribution and a larger proportion of infected cells. The cells most commonly infected with PCV in brain and spinal cord were large neurons. In nonneural tissues, macrophages were the most frequent cell type infected. PCR analysis demonstrated only PCV type 2 and not PCV type 1 in all PCV-infected pigs on all 4 farms.

Viral replication and lesions in BALB/c mice experimentally inoculated with porcine circovirus isolated from a pig with postweaning multisystemic wasting disease.

Eight-week-old BALB/c mice were either sham inoculated (control mice) or were inoculated intraperitoneally (IP) and intranasally (IN) with a single (sPCV mice) or multiple (mPCV mice) doses of porcine circovirus 2 (PCV2). Four control mice and 4 sPCV mice were sacrificed 7, 14, 28, and 42 days postinoculation (PI). All 4 mPCV mice were sacrificed 42 days PI. In addition, 7-day and 14-day pregnant BALB/c mice were either sham inoculated (control mice) or were inoculated IP and IN with a single dose of PCV2. Newborn mice were euthanatized 1, 8, and 15 days after birth. Necropsies were performed on all euthanatized mice and tissues were collected for histopathology, electron microscopy, in situ hybridization, and polymerase chain reaction (PCR). PCV2 replicated in 8-week-old BALB/c mice that were inoculated with PCV2 and caused fetal infection when inoculated into pregnant BALB/c mice at 7 days and 14 days of gestation. PCV was detected by in situ hybridization and PCR in sPCV mice on days 7, 14, 28, and 42 PI; in mPCV mice on day 42 PI; and in newborn mice from mothers inoculated with PCV at 7 days and 14 days of gestation at 1, 8, and 15 days after birth, but not in control mice. No clinical signs or gross lesions were found in sPCV or mPCV mice during the study. Microscopic lesions in sPCV mice and mPCV mice were characterized by expansion of germinal centers in lymphoid organs with large numbers of histiocytic cells and lymphoblasts, apoptosis of histiocytic cells in germinal centers, and mild lymphoid depletion of the paracortex. PCV nucleic acid was detected in the nuclei and cytoplasm of histiocytes and apoptotic cells in germinal centers in lymphoid tissues as well as in the nuclei of hepatocytes in the liver, in the nuclei of renal tubular epithelial cells, and in the cytoplasm of single lymphocytes in the thymus. Congenitally infected mice only had PCV nucleic acid detected in putative Kupffer cells in livers.

Meningoencephalitis in a Boelen's python (Morelia boeleni) associated with paramyxovirus infection.

An adult male Boelen's python, Morelia boeleni, presented with acute neurologic disease and was euthanatized. Histologic examination revealed nonsuppurative meningoencephalitis. Occasional eosinophilic intracytoplasmic inclusions were noted in glial cells. On the basis of clinical signs and histopathology, inclusion body disease of boid snakes was suspected, but inclusions were not seen in other organs commonly affected with the disease. Moreover, electron microscopy revealed that the inclusions contained stacks of filaments 13-14 nm wide. With the use of a generic paramyxovirus cDNA probe, sections of brain and esophageal ganglion demonstrated hybridization. The findings indicate that paramyxovirus was the likely cause of the encephalomyelitis in this python, and this virus should be included in the differential diagnosis of pythons exhibiting central nervous system disease.

Severe leukopenia and liver necrosis in young African grey parrots (Psittacus erithacus erithacus) infected with psittacine circovirus.

This paper describes the signs, clinical pathology, and postmortem findings in 14 young African grey parrots (Psittacus erithacus erithacus) that were naturally infected with psittacine beak and feather disease (PBFD) virus (psittacine circovirus). All but two of the parrots had severe leukopenia at clinical presentation. Two other parrots also had severe anemia. All birds died within 3 wk after presentation. Postmortem examination documented liver necrosis in 11 of 14 birds and secondary bacterial or fungal infections in 9 of 14 birds. Tests for Chlamydia psittaci, polyomavirus, and Salmonella sp. were negative. PBFD viral infection could be demonstrated in all birds by polymerase chain reaction. Supporting evidence of PBFD viral infection was gathered by histologic examination of the bursa of Fabricius, electron microscopy, and DNA in situ hybridization. Electron microscopic examination of both the bursa of Fabricius and liver revealed virus particles resembling circovirus. DNA in situ hybridization of six liver tissue samples confirmed the presence of PBFD virus and excluded the presence of avian polyomavirus. Our findings suggest that a specific presentation of peracute PBFD viral infection, characterized by severe leukopenia, anemia, or pancytopenia and liver necrosis in the absence of feather and beak abnormalities, may occur in young African grey parrots.

Proliferative pododermatitis associated with virus-like particles in a northern gannet.

Small multifocal lesions of proliferative pododermatitis were observed in an emaciated adult male northern gannet (Morus bassanus). Ultrastructurally, these lesions were associated with numerous virus-like particles with a size and morphology suggestive of Papovaviridae. DNA in situ hybridization with probes for avian polyomaviral and papillomaviral nucleic acid and an immunohistochemical test for the presence of papillomaviral antigen failed to identify this virus further. To our knowledge, papovavirus-like particles have not been recognized previously in this avian species.

Pathological, immunohistochemical, and in-situ hybridization studies of natural cases of postweaning multisystemic wasting syndrome (PMWS) in pigs.

Fifteen pigs from five farms on which there had been a previous clinical and histopathological diagnosis of postweaning multisystemic wasting syndrome (PMWS) were investigated. At necropsy, enlargement of lymph nodes was the most obvious lesion; other lesions were non-collapsed lungs, ulceration of the gastric pars oesophagica, and cranioventral pulmonary consolidation. Microscopical lesions attributable to PMWS were found in lymphoid organs (including lymph nodes, tonsil, Peyer's patches and spleen), liver, kidney and lungs. Varying degrees of lymphocellular depletion, affecting both lymphoid follicles and parafollicular zones, and progressive multifocal to diffuse infiltration of lymphoid tissue by large histiocytic cells were the characteristic lesions. Syncytial cells were seen frequently, especially in lymphoid organs. A prominent finding was the presence of sharply demarcated, spherical, basophilic, cytoplasmic inclusions in histiocytic cells. The lymphoid lesions were suggestive of immunosuppression. Non-lymphoid lesions included interstitial pneumonia, periportal mononuclear inflammatory infiltration of the liver in varying degrees, and interstitial nephritis. Porcine circovirus (PCV) antigen and nucleic acid were regularly found in lymphoid organs, lung, liver and, to a lesser degree, kidney. Target cells for PCV replication included monocyte/macrophage lineage and antigen-presenting cells. To a lesser extent, epithelial cells such as renal tubular, bronchial and bronchiolar cells, endothelial cells, hepatocytes and lymphocytes were also labelled. One pig did not show PCV nucleic acid; sequence differences among different viral isolates are discussed as the probable cause of this lack of labelling by the in-situ hybridization PCV-specific probe.

Granular cell tumour in an endangered Puerto Rican Amazon parrot (Amazon vittata ).

A 3 cm diameter mass from the metacarpus of a Puerto Rican Amazon parrot was diagnosed as a granular cell tumour based on light microscopy. The cytoplasmic granules were periodic-acid Schiff positive and diastase resistant. Ultrastructural characteristics of the cells included convoluted nuclei and the presence of numerous cytoplasmic tertiary lysosomes. This is only the second granular cell tumour reported in a bird. We speculate that most granular cell tumours are derived from cells that are engaged in some type of cellular degradative process, creating a similar morphologic appearance, but lacking a uniform histogenesis.

Colonic adenocarcinoma in a corn snake (Elaphe guttata guttata).

Colonic adenocarcinoma was diagnosed by surgical biopsy in a domestically raised 3-yr-old male corn snake (Elaphe guttata guttata). The snake presented with a history of constipation. Several masses were palpated in the distal coelomic cavity. On proctoscopy, a nodular firm white mass encircled the distal colon proximal to the cloaca. The histologic diagnosis was transmural mucinous colonic adenocarcinoma with scirrhous reaction. Resection of the affected region alleviated intestinal obstruction for at least 4 mo, after which the snake was lost to follow-up.

Beak and feather disease virus and porcine circovirus genomes: intermediates between the geminiviruses and plant circoviruses.

Circoviruses are a diverse group of animal and plant pathogens with undefined relationships to one another but for their non-geminate, non-enveloped capsids and circular, single-stranded DNA genomes. The sequences of the beak and feather disease virus and porcine circovirus genomic DNAs are presented and analyzed in the context of the other members of the family. Sequence comparisons, inferred phylogenies, and geographic occurrence suggest that the ambisense circoviruses, particularly the beak and feather disease virus, represent an evolutionary link between the geminiviruses and the plant circoviruses. We propose that the family members be reclassified into three groups: The family Circoviridae consists of the animal pathogens (beak and feather disease virus and porcine circovirus) that possess ambisense genomes with striking similarities to the geminiviruses. The BBTV-like viruses include the plant pathogens (coconut foliar decay virus, banana bunchy top virus, subterranean clover stunt virus) with a geminivirus-like stem-loop element in their DNAs, and single to multiple component genomes. The chicken anemia virus is an unassigned virus possessing unique characteristics bearing little similarity to the other ssDNA viruses.

Axillary cystadenocarcinoma in a Moluccan cockatoo (Cacatua moluccensis).

An adult Moluccan cockatoo (Cacatua moluccensis) was diagnosed with a cystadenocarcinoma in the right axillary region that was treated symptomatically with surgical debulking and periodic drainage. The bird eventually died and a necropsy was performed. The neoplasm extended through the humerus, and small neoplastic foci were seen within the ipsilateral lung parenchyma. Rare groupings of microvilli were observed lining intercellular canalicular lumens on electron microscopy within the axillary tumor. These findings suggest a respiratory neoplasm, although the tissue of origin remained undetermined.

Sarcocystis-associated encephalitis and myocarditis in a wild turkey (Meleagris gallopavo).

A Sarcocystis-like organism was associated with encephalitis and myocarditis in an ataxic, emaciated adult male turkey (Meleagris gallopavo) from Gilmer County, West Virginia. Protozoal schizonts and merozoites were associated with areas of inflammation and occasionally necrosis in both the heart and the brain. The organisms divided by endopolygeny and stained positively with anti-Sarcocystis cruzi serum in an immunohistochemical test.

Use of an inactivated virus vaccine to control polyomavirus outbreaks in nine flocks of psittacine birds.

Nine flocks of psittacine birds were examined because of sudden death of neonates. In each flock, cause of death was determined to be polyomavirus infection, by means of DNA testing and in situ hybridization. Contaminated areas of aviaries were cleaned and disinfected, and vaccination programs, using a recently approved inactivated polyomavirus vaccine, were instituted. Use of the vaccine was found to be safe and efficacious.

Safety, immunogenicity, and efficacy of an inactivated avian polyomavirus vaccine.

OBJECTIVE: To determine safety, immunogenicity, and efficacy of an inactivated avian polyomavirus vaccine in nonbudgerigar psittacine birds that varied in age, species, and immunologic status. ANIMALS: Safety of the vaccine was evaluated in 1,823 psittacines representing more than 80 species. Immunogenicity was evaluated in 285 birds (260 of various Psittaciformes species, 25 chickens). Efficacy was evaluated in 104 birds (78 of various Psittaciformes species, 26 chickens). PROCEDURES: Safety was evaluated by vaccinating birds that were determined to be seronegative or seropositive (titer > 1:10) prior to vaccination. Birds were then evaluated for clinically detectable systemic or local reactions for 2 months to 2 years. Immunogenicity was evaluated by testing for virus-neutralizing antibodies, vaccinating each bird twice, and then testing for a significant change in antibody titer. Efficacy was evaluated by vaccinating birds, followed in 2 to 4 weeks by intramuscular or intravenous challenge exposure. After challenge exposure, protection was evaluated by attempting to recover virus from tissues or by observing birds for clinical signs of disease and testing for a significant change in titer. CONCLUSIONS: Avian polyomavirus vaccine is safe, immunogenic, and efficacious for use in multiple species of mature and immature psittacines. CLINICAL RELEVANCE: Until now, prevention of polyomavirus infection in psittacine birds could only be accomplished through strict isolation to reduce potential exposure to the virus. The USDA-registered inactivated avian polyomavirus vaccine can safely be used to protect vaccinates from infection and control spread of this virus in flocks.