Detection of Hepatitis E Virus Antibodies in Dogs in the United Kingdom.

Hepatitis E virus (HEV) genotypes 3 and 4 are zoonotic pathogens, with pigs predominantly implicated in disease transmission. The rapid rise in human cases in developed countries over the past decade indicates a change in epidemiology of HEV, and it has been suggested that additional animal species may be involved in transmission of infection. Multiple studies have identified contact with dogs as a risk factor for HEV infection in industrialised nations, and a low seroprevalence to HEV has previously been reported in dogs in low-income countries. In this study we aimed to evaluate the possibility that dogs are susceptible to HEV, and determine the frequency with which this occurs. Serum samples from UK dogs with and without hepatitis were screened for HEV-specific antibodies, and canine liver and stool samples were analysed by qPCR for the presence of HEV RNA. We describe evidence to show HEV infection occurs at low levels in dogs in the UK, but the strain of origin is undetermined. The low seroprevalence level of HEV in dogs implies the risk of zoonotic disease transmission is likely to be limited, but further investigations will be required to determine if HEV-infected dogs can transmit HEV to man.This project was made possible by a Wellcome Trust Vacation Scholarship to AM, and was supported by a PhD studentship from the Medical Research Council to SC, a Wellcome Trust Research Training Fellowship to NB (Ref: WT088619MA) and a Wellcome Trust Senior Fellowship to IG (Ref: WT097997MA).This is the final version of the article. It first appeared from PLOS via http://dx.doi.org/10.1371/journal.pone.012870

Canine idiopathic chronic hepatitis

Canine idiopathic hepatitis is a common disease, categorized histologically by presence of hepatocellular apoptosis or necrosis, a variable mononuclear or mixed inflammatory cell infiltrate, regeneration and fibrosis. Clinical signs are vague and non-specific, but there are known breed, age and gender predispositions. Results of clinical pathology are non-specific, but usually include elevations in liver enzymes and function impairment; a liver biopsy is required for diagnosis. Management involves around the use of an anti-inflammatory dose of glucocorticoids and other supportive and symptomatic therapies including ursodeoxycholic acid, antioxidants, diuretics, and diet. The prognosis is variable, but there are known prognostic indicators, especially the presence of portal hypertensio

A Viral Discovery Methodology for Clinical Biopsy Samples Utilising Massively Parallel Next Generation Sequencing

Here we describe a virus discovery protocol for a range of different virus genera, that can be applied to biopsy-sized tissue samples. Our viral enrichment procedure, validated using canine and human liver samples, significantly improves viral read copy number and increases the length of viral contigs that can be generated by de novo assembly. This in turn enables the Illumina next generation sequencing (NGS) platform to be used as an effective tool for viral discovery from tissue samples

Description of neurological mimics presented to the neurology service of a referral small animal hospital

ABSTRACT Background: Clinicians observe that cats and dogs referred to neurology services often do not have an underlying neurological disorder. There has been no analysis of the frequency or categorisation of these neurological mimics. Methods: Retrospective study of 520 cases. Data on signalment, presenting clinical signs, neurological examination findings and final diagnosis was collected. Final diagnoses were classified as primary neurological, non-neurological in origin but with neurological clinical manifestation, completely non-neurological (neurological mimics) or undiagnosed. Presenting clinical signs and neurological examination results were compared between neurological mimics and primary neurological cases using Chi-square or Fischer exact test. Relative risk was calculated for significant associations. Results: 74% were primary neurological conditions, 8% neurological mimics, 3% non-neurological with neurological manifestation and 15% undiagnosed. An animal referred for lameness was five approximately times more likely to be diagnosed as a neurological mimic than as a primary neurological disorder (Relative Risk, RR = 5.42, P < 0.001). Cases with a normal neurological examination were approximately 15 times more likely to be a neurological mimic (RR=14.97, P < 0.001). Conclusion: Thorough examination with consideration of alternative diagnoses is important when a neurological condition is suspected in an animal that presents with lameness or normal neurological examination

Anti-HEV antibody titres in positive canine serum samples.

<p>Positive canine serum samples were prepared in dilutions of 1:100, 1:200, 1:400, 1:800; 1:1600 and 1:3200 and used in an ELISA assay. The corrected OD450 was obtained by subtracting the background signal from the VLP coated well OD450 value. The positive threshold was determined by calculating the mean OD450 of buffer coated wells with the highest serum dilution, plus 3 standard deviations.</p

Western blot analysis of serum sample reactivity with HEV, vesivirus 2117 and human norovirus G1.1 VLPs.

<p>Three types of VLP were separated by SDS-PAGE. One gel was stained with Coomassie Blue to identify VLP protein at the expected molecular weight. Additional gels were used for western blotting with canine serum samples positive by ELISA for HEV (samples A and B). A pig serum sample (kind gift from S. Emerson) and a human serum sample known to be positive for anti-HEV antibody were used as a positive control for the HEV VLPs. Canine sample C, previously confirmed positive for anti-vesivirus antibody by ELISA, was used as a positive control for the vesivirus VLPs. Canine sample D was used as a negative control for all VLPs.</p

Splenectomy in the management of primary immune-mediated hemolytic anemia and primary immune-mediated thrombocytopenia in dogs.

Funder: Linnaeus Veterinary LimitedBACKGROUND: Current reports about the use of splenectomy for the management of immune-mediated hemolytic anemia (IMHA) or immune-mediated thrombocytopenia (ITP) or both in dogs are limited. OBJECTIVES: To retrospectively describe the use of splenectomy as part of the management for IMHA, ITP, and concurrent IMHA and severe thrombocytopenia (CIST) in dogs. It was hypothesized that splenectomy would be beneficial in allowing for reduction of dose of immunosuppressive drugs or discontinuation in 1 or more of these groups. ANIMALS: Seventeen client-owned dogs (7 with IMHA, 7 with ITP, and 3 with CIST) were identified across 7 UK-based referral hospitals from a study period of 2005 to 2016. METHODS: Data were collected retrospectively via questionnaires and included information about diagnosis, management and treatment response before and after splenectomy. Based on clinical outcome, treatment with splenectomy as part of the management protocol was classified as either successful or unsuccessful. RESULTS: Six of 7 dogs with ITP were managed successfully with splenectomy as part of their management protocol (3 complete and 3 partial responses), although 1 subsequently developed suspected IMHA. Of the 7 dogs with IMHA, splenectomy was part of a successful management protocol in 4 dogs (2 complete and 2 partial responses). In the CIST group, 1 case (1/3) responded completely to management with splenectomy as part of the management protocol. CONCLUSIONS AND CLINICAL IMPORTANCE: Splenectomy was considered successful and well tolerated in most cases of isolated ITP. Whether there is a benefit of splenectomy in cases of IMHA and CIST could not be determined in the current study

A multicenter retrospective study assessing progression of biliary sludge in dogs using ultrasonography.

BACKGROUND: Biliary sludge (BS) frequently is identified on ultrasonographic examination and is described as incidental. It is hypothesized that biliary stasis and hypersecretion play a role in both BS and gallbladder mucocele (GBM) formation. Recent studies have documented similarities in composition of BS and GBM, and there are several examples of progression from BS to GBM in the veterinary literature. OBJECTIVES: To assess the relationship between the presence of BS and later development of GBM in dogs, over time periods >12 months. ANIMALS: A total of 154 dogs with BS and ultrasonographic follow-up >12 months. METHODS: Medical records were retrospectively collected from 9 UK-based referral centers for all available time points. A semiobjective scoring system was used to track volume of BS within the gall bladder (GB) over time. RESULTS: Twenty dogs developed GBM during the study period. Shetland Sheepdogs (odds ratio [OR], 40.99; 95% confidence interval [CI], 3.61-465.95; P = .003) and Border Terriers (OR, 11.66; 95% CI, 3.28-46.63; P < .001) were independent risk factors for the development of GBM. Non-gravity-dependent BS (NDBS) was noted to form before GBM development in 9/20 dogs, and breeds at-risk for GBM were more likely to have NDBS. Odds for the development of GBM increased with BS score. CONCLUSIONS AND CLINICAL IMPORTANCE: Dogs with NDBS may be at risk for the development of GBM and a stratified BS scoring system could allow for semiobjective monitoring over time, particularly in at-risk breeds

Quantitative PCR (viral genome copy number in liver samples).

<p>A SYBR-green assay and plasmid standard was used for the CAV and CPV samples, whilst the HCV and HBV assays used dual labelled probed with NIBSC standards. Total RNA extracted for HCV and Total DNA extracted for HBV, CPV and CAV. The Genomic strand copy number for HCV was estimated by performing the RT step in the presence of the reverse primer only.</p

NGS of HCV biopsy RNA.

<p>Extracted and pooled (×6) HCV infected biopsy RNA was reverse transcribed and amplified prior to Illumina NGS and mapped to the HCV reference 3a.NZL. NC_0009824 (Los Alamos HCV database).</p