Sheena M Warman

Development and use of real-time PCR to detect and quantify Mycoplasma haemocanis and “Candidatus Mycoplasma haematoparvum” in dogs

Two canine haemoplasma species have been recognised to date; Mycoplasma haemocanis (Mhc), which has been associated with anaemia in splenectomised or immunocompromised dogs, and “Candidatus Mycoplasma haematoparvum” (CMhp), recently described in an anaemic splenectomised dog undergoing chemotherapy. The study aim was to develop quantitative real-time PCR assays (qPCRs) incorporating an endogenous internal control to detect Mhc and CMhp and to apply these assays to DNA samples extracted from canine blood collected in Northern Tanzania (n = 100) and from dogs presented to a Trinidadian veterinary hospital (n = 185). QPCRs specific for Mhc and CMhp were designed using 16S rRNA gene sequence data, and each was duplexed with an assay specific for canine glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The assays detected ≤10 copies of a sequence-specific haemoplasma plasmid per reaction and neither assay showed cross-reactivity with 106 copies of the sequence-specific plasmid from the non-target canine haemoplasma species. Nineteen of the 100 Tanzanian samples (19%) were positive for Mhc alone and one (1%) was dually infected. One Trinidadian sample was negative for canine GAPDH DNA and was excluded from the study. Of the 184 remaining Trinidadian samples, nine (4.9%) were positive for Mhc alone, five (2.7%) for CMhp alone, and two (1.1%) dually infected. This is the first report of canine haemoplasma qPCR assays that use an internal control to confirm the presence of amplifiable sample DNA, and their application to prevalence studies. Mhc was the most commonly detected canine haemoplasma species.

Dogs with congenital porto-systemic shunting (cPSS) and hepatic encephalopathy have higher serum concentrations of C-reactive protein than asymptomatic dogs with cPSS

Hepatic encephalopathy (HE) is a cause of significant morbidity and mortality in patients with liver disorders and a wide range of rodent models of HE have been described to facilitate studies into the pathogenesis and treatment of HE. However, it is widely acknowledged that no individual model perfectly mimics human HE and there is a particular need for spontaneous, larger animal models. One common congenital abnormality in dogs is the portosystemic shunt (cPSS) which causes clinical signs that are similar to human HE such as ataxia, disorientation, lethargy and occasionally coma. As inflammation has recently been shown to be associated with HE in humans, we hypothesised that inflammation would similarly be associated with HE in dogs with cPSS. To examine this hypothesis we measured C-reactive protein (CRP) in 30 healthy dogs, 19 dogs with a cPSS and no HE and 27 dogs with a cPSS and overt HE. There was a significant difference in CRP concentration between healthy dogs and dogs with HE (p < 0.001) and between dogs with HE and without HE (p < 0.05). The novel finding that there is an association between inflammation and canine HE strengthens the concept that HE in dogs with cPSS shares a similar pathogenesis to humans with HE. Consequently, dogs with a cPSS may be a good spontaneous model of human HE in which to further examine the role of inflammation and development of HE.

Predicting Outcome in dogs with Primary Immune‐Mediated Hemolytic Anemia: Results of a Multicenter Case Registry

Background Outcome prediction in dogs with immune‐mediated hemolytic anemia (IMHA) is challenging and few prognostic indicators have been consistently identified. Objectives An online case registry was initiated to: prospectively survey canine IMHA presentation and management in the British Isles; evaluate 2 previously reported illness severity scores, Canine Hemolytic Anemia Score (CHAOS) and Tokyo and to identify independent prognostic markers. Animals Data from 276 dogs with primary IMHA across 10 referral centers were collected between 2008 and 2012. Methods Outcome prediction by previously reported illness‐severity scores was tested using univariate logistic regression. Independent predictors of death in hospital or by 30‐days after admission were identified using multivariable logistic regression. Results Purebreds represented 89.1% dogs (n = 246). Immunosuppressive medications were administered to 88.4% dogs (n = 244), 76.1% (n = 210) received antithrombotics and 74.3% (n = 205) received packed red blood cells. Seventy‐four per cent of dogs (n = 205) were discharged from hospital and 67.7% (n = 187) were alive 30‐days after admission. Two dogs were lost to follow‐up at 30‐days. In univariate analyses CHAOS was associated with death in hospital and death within 30‐days. Tokyo score was not associated with either outcome measure. A model containing SIRS‐classification, ASA classification, ALT, bilirubin, urea and creatinine predicting outcome at discharge was accurate in 82% of cases. ASA classification, bilirubin, urea and creatinine were independently associated with death in hospital or by 30‐days. Conclusions and clinical importance Markers of kidney function, bilirubin concentration and ASA classification are independently associated with outcome in dogs with IMHA. Validation of this score in an unrelated population is now warranted.

Systematic evaluation of evidence on veterinary viscoelastic testing Part 1: System comparability

Objective To systematically examine the evidence on system comparability between the thromboelastography and the rotational thromboelastometry viscoelastic point-of-care instruments and to identify knowledge gaps. Design Standardized, systematic evaluation of the literature, categorization of relevant articles according to level of evidence and quality, and development of consensus on conclusions for application of the concepts to clinical practice. Setting Academic and referral veterinary medical centers. Results Medline via PubMed, CAB abstracts, and Google Scholar were searched. A total of 8 relevant articles were chosen, none were in support of the question, 1 was neutral to the question (level of evidence [LOE] 6, Poor), and 7 were in opposition to the question (LOE 3 Good; LOE 6 Good; LOE 6 Fair; LOE 6 Poor). Conclusions Results from the 2 analyzers are not directly comparable and extrapolation of the results from one machine to the other should be avoided. Standardization of the preanalytical variables (eg, blood collection, holding time, and temperature during holding) is strongly recommended. It is recommended that each site create their own “site specific” reference values for each machine and that test samples be compared only to the standardized reference values established at that center. Start-up and consumable costs vary between countries and local comparisons should be performed. Decisions should be made based on the expected use of the machine and if multiple operators will be using it.

Infectious agent screening in canine blood donors in the United Kingdom

OBJECTIVES Transfusion of blood products is an important component of veterinary emergency medicine. Donors must be carefully selected to minimise risk of transmission of blood-borne infectious agents. This study was devised to assess the prevalence of such agents in healthy, non-travelled UK dogs screened as prospective donors. METHODS Ethylenediaminetetraacetic acid blood samples from dogs donating blood between August 2007 and January 2012 were screened by polymerase chain reaction for haemotropic mycoplasmas, Bartonella, Babesia, Leishmania, Ehrlichia and Anaplasma spp. Dogs with positive or inconclusive results underwent repeat polymerase chain reaction testing. RESULTS Four of 262 dogs had positive or inconclusive results at initial screening. Repeat polymerase chain reaction testing in each dog was negative, and none of the dogs developed clinical signs of disease. CLINICAL SIGNIFICANCE The positive results on initial screening may have represented false positives from sample contamination or amplification of non-target DNA. It is also possible that dogs were infected at initial sampling but successfully cleared infection before repeat testing. The low number of positive results obtained suggests that prevalence of these agents in a population of healthy UK dogs is low and that use of blood products is unlikely to represent a significant risk of transmission of these diseases.

Serum haptoglobin concentrations in dogs with liver disease

Dogs with liver disease have been shown to have increased serum C-reactive protein (CRP) concentrations. However, it is unclear whether dogs with liver disease also have increased serum haptoglobin concentrations. The aim of the study was to measure serum haptoglobin concentrations in healthy dogs, hospitalised dogs and dogs with liver diseases. Haptoglobin concentrations were measured in 30 healthy dogs, 47 hospitalised dogs with non-hepatic illness, 46 dogs with congenital portosystemic shunt (cPSS) and 11 dogs with primary hepatopathy. Haptoglobin concentrations were not significantly different between cPSS dogs with and without hepatic encephalopathy (HE), thus all cPSS dogs were considered as one group. Haptoglobin concentrations were significantly different between the remaining groups (P<0.0001). Hospitalised ill dogs had significantly higher haptoglobin concentrations than healthy dogs (P<0.001), dogs with cPSS (P<0.001) and dogs with primary hepatopathy (P<0.05). There was no significant difference between haptoglobin concentrations in healthy dogs, dogs with cPSS and dogs with primary hepatopathy. Haptoglobin concentrations were not significantly increased in dogs with liver diseases or in dogs with cPSS and HE. This is in contrast with the previously reported CRP results. This study demonstrates that liver function should be considered when interpreting haptoglobin concentrations in dogs.

Stakeholder consultation on tracking in UK veterinary degrees: part 1

There is on-going debate regarding whether veterinary students should focus on one (or a small number of) species during their undergraduate training (ie, track). The aims of this study were to: evaluate UK stakeholders’ opinion on partial tracking (whereby students continue to qualify able to practise in all species) and full tracking (students qualify in a limited number of species necessitating restricted registration); and evaluate students’ career aspirations in relation to the UK veterinary professions employment profile. This paper presents the quantitative results of surveys completed by practitioners, students and university staff. The majority of respondents (69.4 per cent) disagreed or strongly disagreed with full tracking, however, there was widespread support for partial tracking (79.0 per cent agreed or strongly agreed). Students favoured partial tracking more so than practitioners (P<0.001). Univariate analysis of demographic factors did not identify differences in opinion regarding tracking within stakeholder groups. Students’ knowledge of the UK veterinary employment profile appeared accurate. However, their career aspiration changed with year of the course, and only final year students’ intentions were aligned with the professions current profile. Qualitative data from these surveys are presented in a second paper and include the advantages, disadvantages and implications of partial and full tracking.

Evaluation of a hand-held point-of-care analyser for measurement of creatinine in cats

Objectives The aim of the study was to evaluate whether a handheld creatinine analyser (StatSensor Xpress; SSXp), available for human patients, can be used to measure creatinine reliably in cats. Methods Analytical performance was evaluated by determining within- and between-run coefficient of variation (CV, %), total error observed (TEobs, %) and sigma metrics. Fifty client-owned cats presenting for investigation of clinical disease had creatinine measured simultaneously, using SSXp (whole blood and plasma) and a reference instrument (Konelab, serum); 48 paired samples were included in the study. Creatinine correlation between methodologies (SSXp vs Konelab) and sample types (SSXpwhole blood vs SSXpplasma) was assessed by Spearman’s correlation coefficient and agreement was determined using Bland–Altman difference plots. Each creatinine value was assigned an IRIS stage (1–4); correlation and agreement between Konelab and SSXp IRIS stages were evaluated. Results Within-run CV (4.23–8.85%), between-run CV (8.95–11.72%), TEobs (22.15–34.92%) and sigma metrics (⩽3) did not meet desired analytical requirements. Correlation between sample types was high (SSXpwhole blood vs SSXpplasma; r = 0.89), and between instruments was high (SSXpwhole blood vs Konelabserum; r = 0.85) to very high (SSXpplasma vs Konelabserum; r = 0.91). Konelab and SSXpwhole blood IRIS scores exhibited high correlation (r = 0.76). Packed cell volume did not significantly affect SSXp determination of creatinine. Bland–Altman difference plots identified a positive bias for the SSXp (7.13 μmol/l SSXpwhole blood; 20.23 μmol/l SSXpplasma) compared with the Konelab. Outliers (1/48 whole blood; 2/48 plasma) occurred exclusively at very high creatinine concentrations. The SSXp failed to identify 2/21 azotaemic cats. Conclusions and relevance Analytical performance of the SSXp in feline patients is not considered acceptable. The SSXp exhibited a high to very high correlation compared with the reference methodology but the two instruments cannot be used interchangeably. Improvements in the SSXp analytical performance are needed before its use can be recommended in feline clinical practice.

Diagnoses of canine distemper virus in puppies

WE would like to report the diagnosis of canine distemper virus (CDV) in two poodle-cross puppies in the south-west of the UK. Both puppies were presented shortly after rehoming (two days and two weeks) with acute onset seizures, mild diarrhoea and, in one case, cough. No ocular signs or dermatological …

Feline pancreatitis: current concepts and treatment guidelines

PANCREATITIS is becoming increasingly recognised as a clinical entity in cats. It has always been notoriously difficult to diagnose. Recently, diagnosis has been aided by newer, more sensitive diagnostic tools; however, no test is 100 per cent sensitive or specific, and pancreatitis therefore remains a diagnostic challenge. This article provides an overview of the anatomy and physiology of the pancreas, the pathophysiology involved in pancreatitis and the associated risk factors and clinical signs. It also presents practical guidance on the diagnosis and management of both mild and severe disease in cats.