A survey of enteric organisms detected by real-time PCR assay in faeces of dogs in Western Australia.

This retrospective observational study reports the enteric organisms detected in dogs in Western Australia that had a faecal PCR (fPCR) submitted to a commercial veterinary laboratory. Of 2025 fPCR results, Clostridium perfringens alpha toxin gene was most frequently detected (87.2%), followed by Campylobacter spp. (37.8%), canine parvovirus (10.5%), Giardia spp. (9.7%), Salmonella spp. (7.0%), canine enteric coronavirus (2.3%), and canine distemper virus (0.3%). C.perfringens alpha toxin gene and Campylobacter spp. were the most common organisms co-detected. There was no statistically significant seasonal variation. Further studies are required to elucidate the role these organisms play in gastrointestinal disease in dogs.

Transfusion practice in Australia: an internet-based survey.

OBJECTIVE: To describe small animal transfusion practices in Australia, including access to blood products and frequency of pre-transfusion compatibility testing and medication administration. METHODS: An online survey was disseminated to target Australian veterinarians treating dogs and cats. Information collected included demographics, sources of blood products, blood storage, recipient compatibility testing and administration of medications pre-transfusion. Associations between the use of compatibility tests and premedications were assessed using the χ2 test. Significance was set at P < 0.05. RESULTS: A total of 199 Australian veterinarians were included; however, there was some attrition of respondents over the course of the survey. The majority of respondents were in general practice (n = 133/199). Access to fresh whole blood was commonly reported for dogs (n = 179/199) and cats (n = 131/198), whereas blood components were less commonly available (canine red blood cells [RBC], n = 52/199 and plasma, n = 157/199; feline RBC, n = 9/198 and plasma, n = 21/198). Most blood was sourced from the pets of owners affiliated with the veterinary clinic (n = 179/196). The respondents who did not blood type or crossmatch dogs were significantly more likely to use premedication than those who did these tests (both comparisons: P < 0.001). Likewise, the respondents who did not blood-type cats were significantly more likely to use premedication (P = 0.003); however, there was no association between crossmatching and using premedication in cats (P = 0.183). CONCLUSION: This is the first survey to describe transfusion practices across a variety of practice types throughout Australia. Future work is needed to determine how representative these results are of current transfusion practices across Australia, and if so, what can be done to optimise them.

Faecal PCR panel results and clinical findings in Western Australian dogs with diarrhoea.

AIM: To describe faecal PCR (fPCR) results and clinical findings of dogs seen at a university teaching hospital for diarrhoea. DESIGN: Retrospective case series (April 2015 to July 2018). PROCEDURE: Data were collected from the hospital electronic medical records. Data extracted included signalment, history, clinical signs, treatment, fPCR panel results, other faecal diagnostic test results and antimicrobial use. RESULTS: One hundred and sixty-eight dogs with diarrhoea had a fPCR panel submitted. Most dogs (115, 68.5%) had diarrhoea of 3 days or less duration. Clostridium perfringens alpha toxin gene was most frequently detected (156, 92.9%) by fPCR, followed by Campylobacter spp. (55, 32.7%), canine parvovirus (CPV) (29, 17.3%), Salmonella spp. (14, 8.3%) and Giardia spp. (9, 5.4%). For the 45 dogs that had a negative point-of-care CPV test, 13 were CPV fPCR positive; some of which were adult dogs with current vaccination status. A total of 94/168 (56%) dogs received antimicrobials at some time during the treatment of diarrhoea. CONCLUSION: Faecal PCR panels can identify dogs with enteric organisms in their faeces that traditional faecal diagnostics may miss, thus contributing additional information to the diagnostic process. Nonetheless, fPCR results should be interpreted in light of the clinical findings, and particular consideration given to avoiding inappropriate use of antimicrobials. This study highlights that testing for C. perfringens alpha toxin gene is not likely to be diagnostically helpful, and that adult dogs with diarrhoea might be identified as CPV positive with PCR testing, despite a negative point-of-care CPV test result and a current vaccination status.

SnakeMap: four years of experience with a national small animal snake envenomation registry.

SnakeMap is a national cloud-based, veterinary snakebite registry. It was designed to prospectively collect data of the clinical circumstances and temporospatial information on cases of snake envenomation in dogs and cats. We herein introduce the project and summarise the data from the first 4 years of SnakeMap. The registry is a veterinary community-based online database allowing case entry from veterinary hospitals across Australia. Registry data comprise hospital characteristics, patient characteristics, envenoming snake type, treatment and outcome variables, including time and geolocation of the snake bite. We present summative information on select key variables from the SnakeMap registry (1 July 2015 to 30 June 2019). Twenty-eight hospitals from 6 states/territories entered 624 cases into the registry, including 419 dogs (67%) and 205 cats (33%). Bite time was available in 216 animals of which 90 (42%) were reported to be bitten in the 3 hours between 03:00 pm and 05:59 pm; median bite to presentation interval was 60 (interquartile range [IQR] 30, 211) minutes in dogs and 95 (IQR 41, 238) minutes in cats. Bites occurred in the owner's yard in 356 dogs (85%) and 53 cats (26%). A snake venom detection kit was used in 172 cases (28%) and antivenom was administered in 523 cases (85%). Most animals (n = 534, 88%) survived to discharge (median hospitalisation of 25 [IQR 16, 62] hours). SnakeMap effectively collects relevant clinical data from dogs and cats with presumed snake bite and provides locally specific information on the epidemiology of snake envenomation in small animals.

Intramyocardial haematoma causing right ventricular outflow obstruction after brown snake (Pseudonaja species) envenomation in a dog.

A 15-month-old, male neutered Staffordshire Bull Terrier cross was presented to its referring veterinarian collapsed and agonal. He was immediately intubated, manually ventilated, and treatment commenced for presumptive snake envenomation with two vials of Tiger/Multi-Brown Snake Antivenom (minimum 7000 units/vial). The dog was transferred to a referral hospital intubated. Additional diagnostics performed following arrival at the referral hospital included a urine snake venom detection kit test, which was positive for brown snake immunotype. Three additional vials of Tiger/Multi-Brown Snake Antivenom (minimum 7000 units/vial) were administered until the dog was extubated and able to stand. Venom-induced consumptive coagulopathy (VICC) was diagnosed based on prolonged clotting times and scleral haemorrhage. Paroxysms of right ventricular outflow tract (RVOT) origin ventricular arrhythmias were treated with lignocaine and sotalol. Four days after presentation, a new-grade IV/VI systolic heart murmur was auscultated, prompting an echocardiogram. An anechoic and compartmentalised mass measuring 43 mm × 19 mm was visualized within the right ventricular wall at the RVOT, immediately adjacent to the pulmonic valve. The mass was causing a RVOT obstruction. Its appearance was suggestive of an intramyocardial haematoma, most likely secondary to VICC. The dog remained cardiovascularly stable, and treatment consisted of supportive care. Recheck echocardiograms at 2 and 7 weeks after discharge revealed progressive improvement of the intramyocardial mass and resolution of the associated heart murmur. Although intramyocardial haematomas are rare, it should be considered as a differential in dogs that develop a newly diagnosed heart murmur and/or cardiac arrhythmia following brown snake envenomation.

Prevalence of Elevated Serum Creatinine Concentration in Dogs Presenting to a Veterinary Academic Medical Center (2010-2014).

BACKGROUND: The epidemiology of kidney disease is not extensively described in dogs. HYPOTHESIS/OBJECTIVES: To better understand the prevalence of elevated serum creatinine concentration in dogs. ANIMALS: Client-owned dogs. METHODS: A retrospective, observational cross-sectional study design was used. We made a dataset of 115,631 hospital visits of all dogs presenting from October 2010 to October 2014. We estimated the prevalence and risk of elevated serum creatinine, defined as >1.6 mg/dL, in evaluated dogs. RESULTS: Of 115,631 visits, 98,693 were outpatient visits and 16,938 were hospital admissions. Among outpatient visits, 9,983 (10.1%) had serum creatinine assessment (4,423 [44.3%] visits were first visits), whereas, among hospital admissions, 12,228 (60.0%) had at least 1 serum creatinine (7,731 [75.6%] admissions were first admissions). The prevalence of elevated serum creatinine concentration in all evaluated dogs was 11.5% (95% CI: 11.0%, 11.9%); 10.2% (95% CI: 9.6%, 10.8%) of inpatients and 12.9% (95% CI: 12.1%, 13.8%) of outpatients had elevated serum creatinine concentration. The relative risk (RR) of elevated serum creatinine concentration was significantly higher in geriatric dogs (outpatient RR 1.45 [95% CI: 1.23, 1.70], inpatient RR 1.43 [95% CI: 1.16, 1.76]) and lower in young dogs (outpatient RR 0.39 [95% CI: 0.26, 0.59], inpatient RR 0.44 [95% CI: 0.32, 0.62]) when compared to the measured population risk. CONCLUSIONS AND CLINICAL IMPORTANCE: When selected for laboratory evaluation, the proportion of dogs presenting to an academic medical center with evidence of kidney injury is high compared to previous reports and might reflect a population of sicker dogs.

Serum 25-hydroxyvitamin D concentrations in dogs - correlation with health and cancer risk.

25-hydroxyvitamin D (25(OH)D) is important in bone health as well as many diseases including cancer. Supplementation may increase responsiveness of cancer cells to chemotherapy. Serum 25(OH)D, intact parathyroid hormone (iPTH) and canine C-reactive protein (c-CRP) were measured in healthy dogs and dogs with haemoabdomen. Regression analysis determined optimal 25(OH)D concentrations. In healthy dogs (n = 282), mean iPTH concentrations correlated inversely (r(2) = 0.88, P < 0.001) to 25(OH)D concentrations. Variation in both iPTH and c-CRP plateaued at 25(OH)D concentrations of 100-120 ng mL(-1) . Haemoabdomen dogs (n = 63, 43 malignant and 20 benign) had 25(OH)D concentrations ranging from 19.4 to >150 ng mL(-1) . Relative risk of cancer increased with decreasing 25(OH)D concentrations [RR = 3.9 for 25(OH)D below 40 ng mL(-1) (P = 0.0001)]. Serum 25(OH)D concentrations in dogs vary widely, and are influenced by dietary VitD content. Serum vitD measurement can identify dogs for which supplementation may improve health and response to cancer therapy.

Serum thymidine kinase 1 and C-reactive protein as biomarkers for screening clinically healthy dogs for occult disease.

Thymidine kinase (TK1) is a biomarker that correlates well with diagnosis and prognosis in certain canine cancers. Canine C-reactive protein (cCRP) is a widely accepted marker of inflammation correlated with increased risk and severity of various diseases. We evaluated serum TK1 and cCRP concentrations in apparently healthy dogs (n = 360). All dogs were followed up for a minimum of 6 months by health questionnaire. All dogs with cancer were identified using a proprietary dual-biomarker algorithm [termed Neoplasia Index (NI)]. Specificity of positive NI is 0.91 and high positive is 0.98. All-cause mortality was 20% in dogs with elevated cCRP and 3% in dogs with low cCRP. The performance of serum TK1 and cCRP as tools for screening for occult cancer is improved when evaluated together. Serum TK1 and cCRP (unified in the NI) are useful in the screening of occult canine cancer. cCRP is useful in screening for other serious diseases.

Gastric dilatation and volvulus in a brachycephalic dog with hiatal hernia.

A brachycephalic dog was presented with an acute onset of retching and abdominal discomfort. The dog had a chronic history of stertor and exercise intolerance suggestive of brachycephalic airway obstructive syndrome. Radiographs were consistent with a Type II hiatal hernia. The dog was referred and within hours of admission became acutely painful and developed tympanic abdominal distension. A right lateral abdominal radiograph confirmed gastric dilatation and volvulus with herniation of the pylorus through the hiatus. An emergency exploratory coeliotomy was performed, during which the stomach was derotated, and an incisional gastropexy, herniorrhaphy and splenectomy were performed. A staphylectomy was performed immediately following the exploratory coeliotomy. The dog recovered uneventfully. Gastric dilatation and volvulus is a potentially life-threatening complication that can occur in dogs with Type II hiatal hernia and should be considered a surgical emergency.

Endothelin-1 concentrations in bronchoalveolar lavage fluid of cats with experimentally induced asthma.

BACKGROUND: There is a need for biomarkers for diagnosis, therapeutic monitoring, and prognosis for asthma in cats. Endothelin-1 (ET-1) is implicated in the pathogenesis of inflammatory airway diseases in other species but not the cat. OBJECTIVE: To conduct a prospective experimental study to show that experimentally asthmatic cats, but not control cats without airway inflammation, would have increased concentrations of ET in BALF. ANIMALS: Eleven healthy, adult research cats. METHODS: Prospective experimental study. Six healthy cats without airway inflammation were used as controls. Asthma was induced using Bermuda grass allergen (BGA) in 5 cats. Collection of BALF for total nucleated cell and differential counts was performed. The concentration of ET-1 in cell-free BALF samples was determined. Data were analyzed using a Mann-Whitney U-test with P < .05 considered significant. RESULTS: The median [range] BALF total cell numbers, eosinophil numbers, and eosinophil percentages were significantly higher in the cats following experimental induction of asthma (1,870 cells/µL [1,450-3,440], 711 cells/µL [356-1,686] and 38% [20-49]) compared to baseline control parameters (462 cells/µL [239-780], 18 cells/µL [18-62] and 3.5% [0-8]) (P < .01). The median [range] BALF ET concentration was also significantly higher after induction of asthma (1.393 fmol/mL[0.977-2.247]) compared to healthy control cats (0.83250 fmol/mL [0.625-1.038]) (P = .012). CONCLUSIONS AND CLINICAL IMPORTANCE: This study suggests that BAL ET-1 concentration can be used to differentiate normal cats from those with experimentally induced asthma. If the same holds true for cats with naturally developing asthma, BAL ET-1 may prove a useful diagnostic biomarker for asthma.

Plasma inflammatory mediator concentrations at ICU admission in dogs with naturally developing sepsis.

BACKGROUND: Identifying biomarkers to aide in the diagnosis and prognostication of sepsis in dogs would be valuable to veterinarians. OBJECTIVE: To compare plasma inflammatory mediator concentrations among dogs with sepsis, noninfectious systemic inflammatory response syndrome (NSIRS), and healthy dogs. ANIMALS: Dogs with sepsis (n = 22), NSIRS (n = 23), and healthy dogs (n = 13) presenting to the intensive care unit (ICU) at a veterinary teaching hospital. METHODS: Prospective observational study. Clinical parameters were recorded for each dog and plasma tumor necrosis factor (TNF) bioactivity and concentrations of interleukin (IL)-6, CXC chemokine ligand (CXCL)-8 and IL-10 were determined at ICU presentation. RESULTS: Dogs with sepsis and NSIRS were significantly more likely to have measurable TNF activity (sepsis 20/22; NSIRS 19/20; healthy 0/13) and IL-6 concentration (sepsis 12/22; NSIRS 15/23; healthy 2/13), than healthy dogs. Healthy dogs (9/13) were significantly more likely to have measurable plasma IL-10 concentrations than dogs with sepsis (4/19), but not NSIRS (7/20). None of the inflammatory mediators evaluated had optimal sensitivity or specificity for the diagnosis of sepsis. Twelve of 22 dogs with sepsis and 15/23 dogs with NSIRS survived to discharge; none of the measured biomarkers correlated with survival to discharge. CONCLUSIONS AND CLINICAL IMPORTANCE: Sepsis and NSIRS are associated with increased production of the proinflammatory cytokines TNF and IL-6. In addition, sepsis is associated with decreased production of the anti-inflammatory cytokine IL-10. Despite this, plasma TNF, IL-6, CXCL-8, and IL-10 measured at ICU presentation do not appear to be valuable biomarkers to differentiate sepsis from NSIRS, or predict hospital outcome.

Elevated serum thymidine kinase activity in canine splenic hemangiosarcoma*.

Thymidine kinase 1 (TK1) is a soluble biomarker associated with DNA synthesis. This prospective study evaluated serum TK1 activity in dogs presenting with hemoabdomen and a splenic mass. An ELISA using azidothymidine as a substrate was used to evaluate TK1 activity. Sixty-two dogs with hemoabdomen and 15 normal controls were studied. Serum TK1 activity was significantly higher in dogs with hemangiosarcoma (HSA) than in normal dogs (mean ± SEM = 17.0 ± 5.0 and 2.01 ± 0.6, respectively), but not dogs with benign disease (mean ± SEM = 10.0 ± 3.3). Using a cut-off of 6.55 U/L, TK activity demonstrated a sensitivity of 0.52, specificity of 0.93, positive predictive value of 0.94 and negative predictive value of 0.48 for distinguishing HSA versus normal. When interval thresholds of <1.55 and >7.95 U/L were used together, diagnostic utility was increased. Serum TK1 evaluation may help to discriminate between benign disease and HSA in dogs with hemoabdomen and a splenic mass.

Plasma nitrate/nitrite concentrations in dogs with naturally developing sepsis and non-infectious forms of the systemic inflammatory response syndrome.

The aim of this prospective observational study was to evaluate the differences in plasma nitrate/nitrite concentrations between dogs with sepsis and those with non-infectious forms of the systemic inflammatory response syndrome (SIRS). Eighteen dogs with sepsis, 20 dogs with SIRS and 29 healthy control dogs were enrolled. Blood samples were obtained from the dogs within 12 hours of admission to the University of Missouri Veterinary Medical Teaching Hospital (MU VMTH) Intensive Care Unit (ICU) in lithium heparin blood tubes. Plasma nitrate/nitrite concentrations were measured using the Greiss reaction. Plasma nitrate/nitrite concentrations at presentation, clinical parameters, organ dysfunction and in-hospital mortality were compared between groups. Plasma total nitrate/nitrite was significantly greater in the sepsis group compared with the control group (P=0.005) and SIRS group (P=0.037). There was no statistical difference in plasma nitrate/nitrite concentration between the SIRS and control groups (P=0.489). The sensitivity was 66.7 per cent (95 per cent CI, 41 to 87 per cent) and the specificity was 75.5 per cent (95 per cent CI, 61 to 87 per cent) for differentiating dogs with sepsis from dogs without sepsis.

Evaluation of serum NT-pCNP as a diagnostic and prognostic biomarker for sepsis in dogs.

BACKGROUND: There is a need for diagnostic biomarkers that can rapidly differentiate dogs with sepsis from dogs with noninfectious forms of systemic inflammatory response syndrome (NSIRS). OBJECTIVES: To compare serum NT-pCNP concentrations among dogs with various forms of sepsis, NSIRS, and healthy controls and to evaluate the use of serum NT-pCNP for the diagnosis of various forms of sepsis in dogs. ANIMALS: One hundred and twelve dogs including 63 critically ill dogs (sepsis n = 29; NSIRS n = 34) and 49 healthy control dogs. METHODS: Prospective clinical investigation. Serum samples were collected for NT-pCNP measurement from dogs with sepsis or NSIRS within 24 hours of intensive care unit admission or at the time of presentation for healthy dogs. Dogs with sepsis were subclassified based on the anatomic region of infection. Serum NT-pCNP concentrations were compared among sepsis, NSIRS and healthy groups as well as among sepsis subgroups. The area under the curve (AUC), sensitivity, and specificity for identifying dogs with sepsis were determined. RESULTS: Using a cut-off value of 10.1 pmol/L, AUC, sensitivity, and specificity of NT-pCNP for differentiating dogs with sepsis from dogs with NSIRS or healthy control dogs were 0.71 (95% CI, 0.58-0.85), 65.5% (45.7-82.1%), and 89.2% (80.4-94.9%), respectively. Serum NT-pCNP had poor sensitivity for peritoneal sources of sepsis; AUC [0.92 (0.81-1.0)], sensitivity [94% (71-100%)], and specificity [89% (80-95%)] improved when these dogs were excluded. Serum NT-pCNP concentration was not associated with survival in the sepsis group. CONCLUSIONS AND CLINICAL IMPORTANCE: Serum NT-pCNP is a promising diagnostic biomarker for sepsis but is a poor indicator of septic peritonitis.

Characterization of covalent protein conjugates using solid-state 13C NMR spectroscopy.

Cross-polarization magic-angle spinning (CPMAS) 13C NMR spectroscopy has been used to characterize covalent conjugates of alachlor, an alpha-chloroacetamide hapten, with glutathione (GSH) and bovine serum albumin (BSA). The solid-state NMR method demonstrates definitively the covalent nature of these conjugates and can also be used to characterize the sites of hapten attachment to proteins. Three different sites of alachlor binding are observed in the BSA system. Accurate quantitation of the amount of hapten covalently bound to GSH and BSA is reported. The solid-state 13C NMR technique can easily be generalized to study other small molecule/protein conjugates and can be used to assist the development and refinement of synthetic methods needed for the successful formation of such protein alkylation products.

Immunoaffinity techniques applied to the purification of gibberellins from plant extracts.

The use of immunoaffinity columns containing anti-gibberellin (GA) antibodies for the selective purification of GAs in plant extracts is described. GA(1), GA(3), GA(4), GA(5), GA(7), and GA(9) conjugates to bovine serum albumin were synthesized and used to elicit anti-GA polyclonal antibodies (Abs) in rabbits. Protein A purified rabbit serum, containing a mixture of anti-GA Abs, was immobilized on matrices of Affi-gel 10 or Fast-Flow Sepharose 4B. Columns of these immunosorbents retained a wide range of C-19 GA methyl esters, but no C-20 GA methyl esters. Quantitative recovery of C-19 GA methyl esters was achieved from the columns, which, after reequilibration in buffer, could be reused up to 500 times. The immunosorbents were tested by examination of extracts from immature soybean and pea seeds. GAs were initially purified by passing the extracts through DEAE-cellulose and concentrating them on octadecylsilica. The extracts were methylated and further purified on the mixed anti-GA immunoaffinity columns. GAs were detected and quantified as methyl esters or methyl ester trimethylsilyl ethers by gas chromatography-mass spectrometry-selected ion monitoring. GA(7) was found in soybean seeds, 17 days after anthesis, at low levels (8.8 nanograms per gram fresh weight). C-19 GAs were examined in cotyledons, embryonic axes, and testae of G2 pea seeds harvested 20 days after anthesis. High levels of GA(20) and GA(29) were found in cotyledons (3580 and 310 nanograms per gram fresh weight, respectively) and embryonic axes (5375 and 1430 nanograms per gram) fresh weight, respectively). Lower levels of GA(9) were found in cotyledons and embryonic axes (147 and 161 nanograms per gram fresh weight, respectively). GA(9) was the major GA of testae at levels of 195 nanograms per gram fresh weight. Trace quantities of GA(20) and GA(51) were also observed in testae.