AVES POLYOMAVIRUS 1 IN ARA CHLOROPTERA AND ECLECTUS RORATUS WITH DISCLOSURE OF FULL GENOMIC SEQUENCES

Aves polyomavirus 1 (APyV) is the causative agent of acute disease in birds and causes high mortality rates in nestlings. Infections have been reported worldwide in a significant number of caged bird species, such as parrots, caiques, budgerigars, lovebirds, and macaws. However, the number of complete viral sequences available in public databases is scarce, especially those with associated clinical histories. In this study, the clinical, pathological, epidemiological, and molecular characteristics of 2 APyV strains detected in Portugal are described. In the autumn of 2015, a 2-month-old female green-winged macaw (Ara chloroptera) from a small breeder died with signs of dehydration, weight loss, and depression, raising the suspicion of polyomavirus infection. Histopathological analysis revealed lesions compatible with APyV infection, and the presence of polyomavirus in several organs was confirmed by polymerase chain reaction. From the cohabitants tested (n = 14), 1 eclectus parrot (Eclectus roratus), which was more than 1-year-old, was also APyV DNA-positive. The full genomic sequences of the 2 strains were obtained and found identical, suggesting a single introduction in the premises and the occurrence of subsequent infections by the same virus. When compared with sequences of other APyVs available in public databases, high nucleotide similarity percentages were obtained, confirming the close genetic relationship among polyomaviruses worldwide. Interestingly, strain APV7, detected in a white-bellied parrot in 2008 in Japan, was the closest strain to those isolated in this report. Attempts to isolate the virus in eggs and cell lines failed. Phylogenetic analysis was performed by the Bayesian method to determine the phylogenetic classification of the macaw and parrot strains. Both clustered into group V, together with other strains from different bird species with no host or spatial-temporal relationships being observed.     

Gross abnormalities,bacteriology and histological lesions of uteri of dairy cows failing to conceive or maintain pregnancy

AIM: To describe the gross pathology, bacteriology and histopathology of the reproductive tracts of dairy cows that failed to conceive or maintain pregnancy.

METHODS: The reproductive tracts of 105 cows that were not pregnant at the end of the seasonal breeding programme were retrieved following slaughter. The tracts were examined grossly, both horns of the uterine lumen were swabbed for bacteriology, and tissue was collected from each horn and the body of the uterus for histopathology. Grossly enlarged uteri and tracts containing purulent vaginal content were excluded. Histopathology was performed on three sections of uterine tissue from each of three cows in which no gross pathological changes were detected and from which no bacteria were isolated, from three cows in which no gross pathology was detected but from which bacteria were isolated, and from three cows in which gross pathological changes were detected but from which no bacteria were isolated.

RESULTS: Thirty-six (34%) cows had one or more gross lesions which involved the ovary, uterine tube, uterus or vagina. Bacteria were isolated from the uteri of 22 (21%) cows. Isolates included Arcanobacterium pyogenes (n=1), Escherichia coli (n=1), Fusobacterium spp (n=1), Haemophilus somnus (n=5), Streptococcus acidominimus (n=12), S. bovis (n=2), S. uberis (n=1) and S. salivarious (n=1). In only five cows were both gross pathology and bacteria detected. There was no relationship between the isolation of bacteria and the diagnosis of gross pathology of the uterus. There were no differences in the degree of histopathological changes in the uteri from the three groups of cows examined, and lesions present were minor.

CONCLUSIONS: Gross pathological changes and intra-uterine bacteria were found in 34% and 20% of cows, respectively, but the correlation between the two was poor. Histopathological changes were unremarkable, suggesting the bacteriological findings were coincidental, that causative agents of infertility were not present at the time of examination, or that unrelated causes such as nutritional anoestrus may have been responsible for the failure of some cows to conceive.     

Studies on experimental chlamydial mastitis in goat histoenzymology

Two strains of Chlamydia psittaci (one isolated from aborted goat foetus and the other from brain of a buffalo calf that had died of meningoencephalitis) were injected intracisternally into six goats to produce experimental mastitis. Cryostat sections of 7–8 μm thickness, obtained from udder, teat, liver and kidney of infected and control animals were incubated for histoenzymic demonstration of alkaline-(AKPase), acid-(ACPase) and adenosine-tri-(ATPase) phosphatases; lactate-(LDH) and succinate-(SDH) dehydrogenases and for reduced nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-D). Results demonstrated that AKPase and NADPH-D declined while ACPase accumulated in acinar cells of udder while both NADPH-D and ACPase decreased in teat sinus epithelium. Hepatic canaliculi in perilobular areas of liver lobules registered complete absence of AKPase and ATPase. Hepatocytes and renal tubules accumulated LDH, SDH and NADPH-D. The interstitial connective tissue of udder and kidney presented higher levels of AKPase. Comparison of results with biochemical alterations in the level of these enzymes revealed striking discrepancies which seem to arise because of failure of biochemical procedures to discriminate between functional cells of tissue and inflammatory cells. The functional significance of histoenzymic alterations has been discussed.     

Clinicopathological features of 11 suspected outbreaks of bovine adenovirus infection and development of a real-time quantitative PCR to detect bovine adenovirus type 10

CASE HISTORY: A retrospective study was conducted to investigate 11 outbreaks of presumptive fatal adenovirus infection diagnosed through two New Zealand diagnostic laboratories during 2014 and 2015. Outbreaks occurred in 6–12-month-old Friesian or Friesian cross cattle during autumn, winter and spring. Individual outbreaks were short in duration, with mortality rates ranging from 3/250 to 20/600 (1.2 to 3.3%).

CLINICAL AND PATHOLOGICAL FINDINGS: Clinical signs included severe diarrhoea, depression, recumbency, and death. Post-mortem examination revealed congestion and oedema of the alimentary tract and fluid to haemorrhagic intestinal contents. Histopathological lesions were characterised by congestion and haemorrhage of the alimentary tract mucosa, oedema of the submucosa, and mild interstitial inflammation in the kidneys. Large basophilic intranuclear inclusion bodies were identified in vascular endothelial cells of the alimentary tract in 11/11 cases and of the kidney in 8/9 cases.

MOLECULAR TESTING: A real-time quantitative PCR (qPCR) assay was designed to detect bovine adenovirus type 10 (BAdV-10) using hexon gene sequences available in GenBank. DNA extracted from a field case and confirmed by sequencing was used as a positive control. The qPCR had a reaction efficiency of 101% (R²=0.99) and the limit of detection was <10 DNA copies/reaction. The qPCR detected BAdV-10 in formalin-fixed paraffin-embedded (FFPE) tissue from 10/11 cases. DNA sequencing of PCR products from nine of these cases showed them to be identical to BAdV-10 sequences in GenBank. For the PCR-negative case, the PCR product had a hexon sequence 99% similar to bovine adenovirus Wic isolate Ma20-1, a close relative of BadV-10.

DIAGNOSIS: Bovine adenovirus type 10 was identified in FFPE tissues from cattle with histopathological evidence of adenovirus infection.

CLINICAL RELEVANCE: Bovine adenoviruses, and especially BAdV-10, should be considered in the differential diagnosis for acute enteric disease and death in young cattle. The qPCR detected BAdV-10 from FFPE tissue of cattle with suspected adenoviral infection diagnosed by histopathology. However results should be interpreted in light of clinical and pathological findings due to the possibility of adenovirus shedding by healthy cattle and the presence of pathogenic adenoviruses other than BAdV-10.     

Multiple mandibular ossifying fibromas in a yearling Belgian Draught horse filly

A yearling Belgian Draught horse filly was presented with a history of bilateral, progressive, painful swelling of the mandible, fever and anorexia. Radiographs and computed tomographic images showed an expansile lesion with a coalescing, well‐defined cystic pattern at the level of the horizontal ramii of the mandible. The histopathological findings confirmed the diagnoses of bilateral, multiple ossifying fibroma. This report indicates that ossifying fibroma should be included in a differential diagnosis if multiple expansile lesions of the jaw are present.     

IMAGING DIAGNOSIS—COMPUTED TOMOGRAPHIC,SURGICAL, AND HISTOPATHOLOGIC CHARACTERISTICS OF AN INFILTRATIVE ANGIOLIPOMA IN A DOG

A 6‐year‐old female spayed Shetland Sheepdog presented for evaluation of a subcutaneous mass over the right prescapular region. The mass had been cytologically diagnosed as a lipoma by the referring veterinarian 20 months prior, but had grown significantly and was very firm. CT scan of the mass was suggestive of neoplasia; however, the tissue of origin could not be determined. Histopathologic evaluation diagnosed infiltrative angiolipoma, and marginal resection of the tumor was performed. Infiltrative angiolipomas are benign but locally aggressive neoplasms uncommonly reported in veterinary medicine. This report correlates the clinical, CT, and histopathologic characteristics of an infiltrative angiolipoma.     

Replacement effect of vitamin E with grape polyphenols on antioxidant status,immune, and organs histopathological responses in broilers from 1- to 35-d age

A study was carried out to investigate the replacement effect of vitamin E with grape polyphenols (GPP) on antioxidant status, immune response, and organs histopathology in broilers from 1- to 35-d age. One-day-old Hubbard broiler chicks (n = 280) were randomly divided into 28 study units of 10 chicks each. A starter control diet was formulated having 22.5% CP, 3,200 kcal/kg ME, and 100 ppm vitamin E50 [50 IU DL-α-tocopheryl acetate (CON)]. Other dietary treatments were obtained by replacing vitamin E on wt:wt ratio, with 25 [low GPP (LGPP)], 50 [medium GPP (MGPP)], and 75 ppm [high GPP (HGPP)] GPP. Likewise, a control finisher diet (20% CP and 3,200 kcal/kg ME) and LGPP, MGPP, and HGPP diets were formulated having the same proportions of GPP and vitamin E as in the respective starter diets. Each dietary treatment was randomly assigned to 7 replicate pens. As a result of this study, by increasing GPP level in place of vitamin E, total phenolic contents in breast and leg muscles increased linearly (P< 0.05) whereas thiobarbituric acid reactive substances decreased quadratically with MGPP showing the lowest value (P< 0.05). Antibody titers against Newcastle disease virus were higher (P< 0.05) in broilers fed the LGPP and HGPP diets than in those fed the control diet. Antibody titers against infectious bursal disease virus were higher (P< 0.05) in broilers fed the HGPP diet than in all of the remaining dietary treatment groups. Histopathological examination of kidney and liver tissues showed no changes in birds fed LGPP and MGPP diets; however, mild effects were observed in some birds fed HGPP diet. Replacement of vitamin E with GPP resulted in lower cost per kg broiler feed. In conclusion, replacement of 75 ppm vitamin E with GPP improved antioxidant status and immune response of broilers at lower feed cost without any significant effect on histopathology of organs.     

Is the use of turmeric in the diet safe for fish?

With this study, we aimed to assess the safety of using Curcuma longa in fish feed. Juvenile Astyanax aff. bimaculatus (0.83 ± 0.04 g) were kept in 24 80‐L aquaria, at a density of 0.5 fish L^?1 for 60 days. Six diets supplemented with 0.0, 20.0, 40.0, 60.0, 80.0 and 100.0 g kg^?1 were evaluated. Histopathological analysis of the gills showed that there was an increasing linear effect of turmeric on the frequency of lamellar aneurysm, hypertrophy of mucous cells, oedema and necrosis. There was a quadratic effect of turmeric supplementation on the frequency of fusion and disruption of secondary lamellae, vacuolation and hypertrophy of the epithelial cells, and the estimated values to minimize these variables were 4.63, 25.93 and 24.46 g kg^?1 respectively. There was a quadratic effect of turmeric on the gills index (I_BRA), and the estimated value to minimize this index was 30.23 g kg^?1. No effect of turmeric on the frequency of histopathological changes and organ index in fish liver was observed. Thus, we conclude that turmeric is not hepatotoxic for A. aff. bimaculatus; however, its use is safe at concentrations up to 30.23 g kg^?1 diet as a function of its gill toxic effect at higher doses.     

Pancreas disease (PD) in sea‐reared Atlantic salmon,Salmo salar L., in Norway; a prospective,longitudinal study of disease development and agreement between diagnostic test results

A prospective longitudinal study was performed on three cages at each of three Norwegian Atlantic salmon seawater sites that experienced outbreaks of pancreas disease (PD). Once salmonid alphavirus (SAV) ribonucleic acid (RNA) was detected by real‐time RT‐PCR (Rt RT‐PCR) at a site, it became detected in all studied cages and was persistently found until the end of the study period up to 19 months after first detection. SAV‐specific antibodies were detected at all sites until the end of the study period and were also found at a high prevalence in broodfish at the time of stripping. No evidence of increased viral activity was detected in these broodfish. One site tested negative over several months prior to the first detection of SAV by Rt RT‐PCR and SAV‐specific antibody, which occurred 1 month prior to clinical manifestations of PD. Moribund fish or thin fish/runts that were sampled after the first PD diagnosis had almost twice the risk of testing positive by one or more diagnostic tests compared to that of randomly selected apparently healthy individuals. This paper describes the first detailed investigation of the disease development of PD at site and cage level in Norway, as well as an assessment of the performance and agreement of the commonly used diagnostic tests.