Early decrease of ionized calcium and static symmetric dimethylarginine concentration in a model of renal tubular necrosis in corn snakes (Pantherophis guttatus).

OBJECTIVE: To determine if plasma concentrations of symmetric dimethylarginine (SDMA), N-acetyl-beta-d-glucosaminidase (NAG), GGT, ALT, AST, lactate, total calcium, and ionized calcium (iCa) and the calcium:phosphorus ratio are clinically relevant biomarkers to detect early stages of tubular lesions in snakes. ANIMALS: 6 adult corn snakes (Pantherophis guttatus). METHODS: Corn snakes were administered 11 injections of gentamicin at 50 mg/kg, SC, q 24 h in an experimental model of induced tubular necrosis. Plasma biochemistry and blood gas analyses were performed at baseline and after the 3rd and 11th injections. Parameters were compared between time points using a paired Wilcoxon test. In 3 individuals, renal biopsies were collected at baseline before starting injections and at the 3rd and 11th injections, while renal tissue samples were procured after euthanasia in all individuals. RESULTS: Renal proximal and distal tubular necrosis and hepatic steatosis were present in all individuals at necropsy. Compared to baseline, decreased blood concentrations of lactate, ionized calcium, and total calcium and a decreased calcium:phosphorus ratio were noted. A significant decrease of lactate and ionized calcium was observed after 3 days. Conversely, no changes in SDMA, NAG, ALT, AST, GGT, and sodium were detected. CLINICAL RELEVANCE: Ionized calcium and lactate concentrations were the earliest parameters to decrease compared to baseline values in this experimental model. While SDMA is a sensitive indicator of renal disease in mammals, this biomarker did not increase in a model of induced acute tubular necrosis in corn snakes.

Determination of Coagulation Parameters by Whole Blood Dynamic Viscoelastic Coagulometry in Strigiformes.

No reference values are available in Strigiformes to evaluate blood coagulation using dynamic viscoelastic coagulometry (DVC) with the Sonoclot (Sienco, Boulder, CO, USA) analyzer. The objectives of this study were 1) to assess the feasibility of DVC in Strigiformes, 2) to calculate the index of individuality of each coagulation parameter, and 3) to assess interspecies variability and establish reference intervals, if relevant, based on the index of individuality. Fresh whole blood samples were obtained from healthy Strigiformes, including 13 barred owls (Strix varia), 10 great horned owls (Bubo virginianus), 6 snowy owls (Bubo scandiacus), and 7 eastern screech owls (Megascops asio), and analyzed with DVC with glass bead (gb) and kaolin clay (k) coagulation activators. Activated clotting time (ACT), clot rate (CR), and platelet function were determined immediately after collection using fresh native whole blood. Intraindividual variability was assessed with a second fresh native whole blood sample from 5 barred owls. Interindividual variability was assessed using a Kruskall-Wallis test. For the parameters gbACT (n = 35), gbCR (n = 34), and kACT (n = 27), no significant differences were detected between species (all P ≥ 0.05). Based on low index of individuality, global Strigiformes reference intervals were determined for gbACT (32.3-852.5 seconds; n = 35), gbCR (0-20.1 units/min; n = 29), and kACT (0-1570.3 seconds; n = 27). In conclusion, DVC can be used in Strigiformes and the gb coagulation activator would be more appropriate when basal individual values are not available in a tested individual.

Establishment of preliminary reference intervals and cytochemical staining of blood cells in big-bellied seahorses (Hippocampus abdominalis).

BACKGROUND: Despite their popularity, hematology reference intervals (RIs) have not been established in big-bellied seahorses (Hippocampus abdominalis). OBJECTIVE: The objectives of this study were to establish hematologic RIs to compare values between sex in regard to cytochemical staining of blood cells. We also sought to compare white blood cell concentrations using the Natt and Herrick technique vs blood smear estimates. METHODS: Forty-three healthy individuals from the Aquarium du Québec (22 females and 21 males) were included. Normal health status was confirmed by an unremarkable physical examination in five individuals and by necropsy of five other individuals, of which all were excluded from further analyses. Venipuncture was performed from the ventral coccygeal vein in the remaining 33 individuals without anesthesia using heparinized insulin syringes. A blood volume of 0.05 to 0.1 ml was collected to prepare Wright Giemsa-stained blood smears and hematocrits immediately after venipuncture. Whole blood was stored in heparinized Eppendorf tubes to determine red and white blood cell concentrations using the Natt and Herrick technique with a hemocytometer in 10 individuals; these results were compared with blood smear estimates. Additional blood smears were stained with alkaline phosphatase substrate, periodic acid Schiff, and toluidine blue stains. RESULTS: The reference intervals included the packed cell volume (27.4-67.5%), thrombocyte count (19.5-197.7 × 109 /L), and white blood cell (WBC) count (2-54.8 × 109 /L), including neutrophils (1.1-21.3 × 109 /L), lymphocytes (2.7-45.5 × 109 /L), and monocytes (0-2.2 × 109 /L). The WBC hemocytometer counts showed no correlation with blood smear estimates (Spearman's rho = 0.2). There was also no significant difference between the sexes. CONCLUSIONS: These preliminary reference intervals will help assess the health of seahorses.

Bile composition of healthy cats and cats with suspected hepatobiliary disease using point-of-care analyzers: A prospective preliminary study.

Although hepatobiliary disease is common in cats, little is known about the bile composition in either these diseased states or in healthy cats. The objectives of this study were to evaluate several analytes from the bile of healthy cats and to investigate the usefulness of measuring these variables to predict bacterial cholangitis. Cats were prospectively enrolled and divided into 3 groups: 21 healthy cats (group 1) and 14 cats with suspected hepatobiliary disease: 9 without bacterial biliary infection (group 2) and 5 with bacterial biliary infection (group 3). Percutaneous ultrasound-guided cholecystocentesis was conducted on each cat. Bile cytology and culture were carried out and bile was analyzed for pH, lactate, and glucose levels using several point-of-care (POC) devices. Reference values for several bile analytes in healthy cats were calculated and are presented in this study. Neither the pH (P = 0.88) nor the lactate concentration (P = 0.85) was significantly different among the 3 groups. Sodium concentration was significantly higher in group 3 than in group 2 (P < 0.05). Bile pH, lactate, and glucose levels were unable to predict the presence of a bacterial infection in the bile.

La composition de la bile est méconnue tant chez les chats sains que chez les chats atteints de maladies hépatobiliaires bien que ces maladies soient fréquentes. Les objectifs de cette étude étaient d'évaluer plusieurs paramètres dans la bile de chats sains et d'investiguer l'utilité de ces derniers comme marqueurs prédictifs de cholangite bactérienne. Les chats ont été recrutés prospectivement et répartis en trois groupes : 21 chats sains (groupe 1); et 14 chats avec maladie hépatobiliaire suspectée : 9 sans infection biliaire (groupe 2) et 5 avec infection bactérienne biliaire (groupe 3). Une cholécystocentèse a été réalisée sur chaque chat. La bile a été utilisée pour cytologie, culture et analyses biochimiques incluant le pH, le lactate et le glucose avec des appareils au chevet du patient. Les valeurs de référence de plusieurs paramètres biliaires chez les chats sains ont été calculées et sont présentées dans cette étude. Le pH (P = 0,88) et la concentration en lactate (P = 0,85) n'étaient pas significativement différents entre les 3 groupes. La concentration en sodium était significativement plus élevée dans le groupe 3 comparée aux groupe 2 (P < 0,05). Les concentrations en lactate et glucose ainsi que le pH ne permettaient pas de prédire une infection bactérienne dans la bile.(Traduit par les auteurs).

Retrospective evaluation of anemia and erythrocyte morphological anomalies in dogs with lymphoma or inflammatory bowel disease.

OBJECTIVE To assess the prevalences of anemia and various RBC anomalies in dogs with lymphoma versus inflammatory bowel disease (IBD) and to evaluate potential relationships between these variables and the severity of lymphoma. DESIGN Retrospective cross-sectional study. ANIMALS 82 client-owned dogs. PROCEDURES Medical records and blood smears were reviewed for dogs in which IBD or lymphoma had been diagnosed between January 1, 2006, and December 31, 2014, and for healthy dogs evaluated during that time frame. Hematologic data were analyzed, and results were compared among groups of healthy dogs, dogs with IBD, and dogs with lymphoma. Results were also compared within the lymphoma group between dogs further grouped on the basis of lymphoma clinical stage, substage, and cell size. RESULTS Prevalence of anemia was significantly higher in dogs with lymphoma (17/32 [53%]) than in dogs with IBD (5/23 [22%]). The total number of different RBC anomalies was significantly higher in dogs with lymphoma than in dogs that were healthy or had IBD. A cutoff of 3 different RBC anomalies/dog enabled differentiation between lymphoma and IBD, with a sensitivity and specificity of 71% and 70%, respectively (area under the fitted curve, 0.7239 ± 0.0727). The presence of eccentrocytes was the only individual RBC anomaly significantly more common in dogs with lymphoma (8/28 [29%]) versus dogs with IBD (1/23 [4%]). CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that detection of anemia combined with ≥ 3 RBC morphological anomalies, particularly eccentrocytes, on blood smears should increase the clinical suspicion of lymphoma, compared with IBD, in dogs.

Granulocytic anaplasmosis in 2 dogs from Quebec.

Two dogs from Quebec were diagnosed with granulocytic anaplasmosis. They both displayed fever, lethargy, and anorexia. Other clinical signs included vomiting, uveitis, polyarthritis, hepatomegaly, and splenomegaly. Thrombocytopenia, anemia, and lymphopenia were identified in both cases. Cytoplasmic inclusions were observed within neutrophils, and Anaplasma phagocytophilum infection was confirmed by polymerase chain reaction in both dogs.

Anaplasmose granulocytaire chez deux chiens au Québec. Deux chiens originaires du Québec ont été diagnostiqués avec une anaplasmose granulocytaire. Les chiens ont manifesté de façon aiguë de la fièvre, un abattement et de l'anorexie. D'autres signes cliniques ont été observés incluant vomissement, uvéite, polyarthrite, hépatomégalie et splénomagalie. Une thrombocytopénie, une anémie et une lymphopénie ont été détectées chez les deux chiens. Des inclusions intracytoplasmques étaient également présentent dans les neutrophiles et l'infection à Anaplasma phagocytophilum a été confirmée par réaction d'amplification en chaîne par la polymérase chez les deux chiens.(Traduit par les auteurs).

Babesia odocoilei as a cause of mortality in captive cervids in Canada.

Nine cases of fatal infection with Babesia odocoilei were confirmed in reindeer (Rangifer tarandus tarandus) and elk (Cervus canadensis) housed in zoological institutions located in southern Quebec, Ontario, and Manitoba, Canada between 2013 and 2016. All animals died of a hemolytic crisis. Frequent postmortem findings were extensive hemorrhage, pigmenturia, and intrahepatic cholestasis. The described ante- and postmortem signs are consistent with those of previously reported cases in the United States. Diagnosis was confirmed in all cases by polymerase chain reaction performed on DNA extracted from whole blood or frozen spleen. We propose that babesiosis is an emerging disease of cervids in multiple Canadian provinces, most likely as a result of climate change and the northward range expansion of Ixodes scapularis, the primary tick vector for B. odocoilei. The role of captive animals as sentinels for wildlife health is also highlighted.

Babesia odocoilei,une cause de la mortalité chez les cervidés captifs au Canada. Entre 2013 à 2016, neuf cas d'infection fatale par Babesia odocoilei ont été détectés chez des caribous (Rangifer tarandus tarandus) et des wapitis (Cervus canadensis) gardés dans des établissements zoologiques situés dans le sud du Québec, de l'Ontario et du Manitoba, Canada. Les animaux sont morts suite à une crise hémolytique. Hémorragies, pigmenturie et cholestase intrahépatique ont fréquemment été identifiées à l'examen postmortem. Les signes ante- et postmortem décrits correspondent avec ceux des cas précédemment signalés aux États-Unis. Le diagnostic de babésiose fut confirmé par réaction en chaîne par polymérase sur l'ADN extrait d'échantillons de sang ou de rate congelée. Nous proposons que la babésiose des cervidés est une maladie émergente au Canada, et ce probablement en conséquence du réchauffement climatique et du mouvement vers le nord de la tique Ixodes scapularis, le principal vecteur de B. odocoilei. La valeur des animaux captifs comme sentinelles pour la santé de la faune est également discutée.(Traduit par les auteurs).

In vitro effects of dalteparin on thrombin generation in canine plasma.

BACKGROUND: The calibrated automated thrombogram (CAT) is a functional thrombin generation (TG) assay that may provide a new approach for monitoring anticoagulant therapy in dogs. The effects of dalteparin on TG variables in dogs are unknown. OBJECTIVES: Objectives were to establish normal TG variable ranges in dogs and measure the in vitro TG variables in canine pooled platelet-poor plasma (PPP) spiked with different dalteparin concentrations. METHODS: In the first experiment, plasma samples from 25 adult healthy Beagle dogs and 11 client-owned healthy dogs of multiple breeds was measured individually for obtaining normal TG values. In the second experiment, separate pools of the remaining PPP from 24 of the 25 previous adult Beagles and from 45 different client-owned dogs were spiked with dalteparin at 9 concentrations with increasing anti-factor Xa (anti-FXa) activity. Activated partial thromboplastin time, tissue factor-induced TG, and anti-FXa activity were measured for each concentration. Concentration-response relationships were determined with ADAPT v.5, using various nonlinear regression models for stimulatory or inhibitory effects. RESULTS: Thrombin generation ranges of client-owned dogs and Beagles were equivalent only for time-to-peak (P < .05). In vitro dalteparin resulted in a concentration-dependent decrease in endogenous thrombin potential (ETP) in pooled PPP. The estimated dalteparin concentration that produced half the maximal inhibition of baseline ETP (IC50 ) was 0.289 U/mL. Thrombin generation and anti-FXa activity were more sensitive than APTT to detect the effects of dalteparin. CONCLUSIONS: The CAT assay can measure the effects of dalteparin in canine plasma, resulting in significant dose-dependent decreases in ETP, prompting further in vivo investigation.

Effect of dalteparin administration on thrombin generation kinetics in healthy dogs.

BACKGROUND: Dalteparin is used to prevent thrombotic complications in dogs. Measurement of anti-factor Xa (anti-FXa) activity is currently used for monitoring therapy, but remains a nonfunctional test. The calibrated automated thrombogram (CAT) could be a suitable approach for functional monitoring. OBJECTIVES: We hypothesized that the CAT will detect decreased endogenous thrombin potential (ETP) in healthy dogs receiving dalteparin. METHODS: Twenty-four healthy adult Beagles were randomly allocated to 4 equal groups. A single subcutaneous (SC) dose of 50 U/kg, 100 U/kg, or 150 U/kg of dalteparin was given. Platelet-poor plasma (PPP) was collected over a 24-hour period and evaluated by thrombin generation (TG) via CAT, anti-FXa activity, and APTT. Analysis was performed with a repeated-measures general linear mixed model, and the treated groups were compared to a placebo group. RESULTS: Time, dose, and time-dose interaction significantly affected ETP (P < .0001 for all effects), peak (P < .0001 for all effects), rate index (P < .0006 for all effects), and anti-FXa activity (P < .0001 for all effects). No significant time trend was detected in the control group. Dogs receiving the 100 U/kg dalteparin SC injection showed the most homogeneous response of ETP inhibition among treated groups. The % inhibition of ETP from baseline increased nonlinearly as a function of anti-FXa activity (r2 = .8186). CONCLUSIONS: The CAT assay can be employed to measure the effects of dalteparin at different doses in healthy dogs, showing sensitivity to time- and dose-dependent changes in ETP and other TG variables. Further investigation of the CAT as a tool for monitoring low molecular weight heparin therapy in dogs is warranted.

Diffuse leptomeningeal histiocytic sarcoma in the cerebrospinal fluid of 2 dogs.

Two adult male castrated dogs were evaluated for progressive paraparesis and ataxia. Neurologic examination showed severe ataxia, delayed proprioceptive placement in the pelvic limbs, pain upon palpation of the lumbar spine as well as facial paresis in one dog, and decreased withdrawal reflex of the pelvic limbs in the other dog. Magnetic resonance imaging (MRI) in both dogs showed diffuse meningeal and intramedullary lesions. However, no evidence of a mass was found. Biopsies could not be performed safely due to the location of the lesions. Cerebrospinal fluid (CSF) examination revealed an inflammatory pleocytosis associated with increased protein concentration and numerous large atypical round cells, often multinucleated. Nuclear fragmentation, micronuclei, and rare atypical mitoses were observed. Immunocytochemistry revealed CD1(+) and CD11c(+) staining, which, in concert with the morphology confirmed the diagnosis of histiocytic sarcoma (HS). Euthanasia was elected due to poor prognosis. Histopathologic examination showed diffuse spinal and meningeal infiltration with CD18(+) neoplastic cells, without any evidence of mass formation, which completed the diagnosis of diffuse leptomeningeal HS involving the brain and the spinal cord. Canine central nervous system (CNS) HS has been seldom reported in the literature, with only isolated cases identified on CSF cytology. The cases reported here are remarkable in describing a diffuse CNS leptomeningeal HS associated with neoplastic cells in the CSF of dogs without a tumor mass. These cases emphasize the potential critical importance of CSF analysis in providing an antemortem diagnosis of neoplasia in neurologic patients.

Cytological analysis of bronchoalveolar lavage fluid acquired by bronchoscopy in healthy ferrets: A pilot study.

The objective of this study was to investigate the normal cytological evaluation of bronchoalveolar lavage (BAL) fluid in healthy adult ferrets (N = 12). These ferrets underwent bronchoscopy and BAL using sterile saline [1.5 mL/kg body weight (BW)]. Percentage of fluid recovered, total leukocyte count, differential leukocyte count, and cell count of the epithelial lining fluid (ELF) were determined. The mean percentage of lavage volume recovered from the right lung and left lung were 67.8 ± 14.9% and 69.7 ± 20.0%, respectively. Gender (P = 0.12) and weight (P = 0.17) did not significantly affect the mean percentage of recovered volume. The mean percentage of recovered volume (P = 0.47) and the mean leukocyte count (P = 0.17) from the right and left lung were not significantly different. Macrophages were the main leukocyte component of the lavages, followed by neutrophils, lymphocytes, and eosinophils. The mean proportion of ELF in BAL fluid was 9.3 ± 3.7% v/v. Bronchoscopy is clinically useful for collecting good quality BAL samples for cytological analysis in ferrets. The leucocyte differential was established, which may help veterinarians to make better clinical decisions when treating respiratory disease. Further studies are required with a larger group in order to establish the healthy reference intervals for BAL values in ferrets.

L'objectif de la présente étude était d'examiner l'évaluation cytologique normale de fluide de lavage bronchoalvéolaire (LAB) de furets adultes en santé (N = 12). Ces furets ont subi ne bronchoscopie et un LAB avec de la saline stérile [1,5 mL/kg de poids corporel (PC)]. Le pourcentage de fluide recouvré, le nombre total de leucocytes, un comptage différentiel de leucocytes, et un compte cellulaire du liquide sécrété par l'épithélium respiratoire ont été déterminés. Les pourcentages moyens de volume de lavage recouvrés du poumon droit et du poumon gauche étaient de 67,8 ± 14,9 % et 69,7 ± 20,0 %, respectivement. Le sexe (P = 0,12) et le poids (P = 0,17) n'ont pas affecté significativement le pourcentage moyen du volume recouvré. Le pourcentage moyen de volume recouvré (P = 0,47) et de comptage leucocytaire moyen (P = 0,17) provenant du poumon droit et du poumon gauche n'étaient pas significativement différents. Les macrophages étaient les principaux leucocytes retrouvés dans les lavages, suivis par les neutrophiles, lymphocytes et éosinophiles. La proportion moyenne d'ELF dans le fluide de LBA était de 9,3 ± 3,7 % v/v. La bronchoscopie est utile cliniquement pour prélever des échantillons de LAB de bonne qualité pour l'examen cytologique chez les furets. Le différentiel des leucocytes a été établi, ce qui pourrait aider les vétérinaires à prendre de meilleures décisions cliniques lors du traitement de maladies respiratoires. Des études supplémentaires sont requises avec un plus grand groupe afin d'établir les intervalles des valeurs de référence des LAB de furets en santé.(Traduit par Docteur Serge Messier).

Effect of castration on the urinary protein-to-creatinine ratio of male dogs.

OBJECTIVE: To assess the urinary protein-to-creatinine ratio (UPCR) of healthy sexually intact male dogs and to compare the UPCR of these dogs before and after castration. ANIMALS: 19 client- or shelter-owned healthy adult sexually intact male dogs. PROCEDURES: Physical, hematologic, and biochemical examinations and urinalysis (including calculation of the UPCR) were performed on each dog. Dogs were then castrated, and physical examination and urinalysis (including calculation of the UPCR) were performed again at least 15 days after castration. RESULTS: A dipstick test yielded positive results for protein in the urine of 10 sexually intact male dogs, but the UPCR was < 0.5 for all sexually intact male dogs. Mean UPCR for sexually intact male dogs was 0.12 (range, 0.10 to 0.32). The UPCR was < 0.2 for all castrated dogs, except for 1. Mean UPCR for all castrated dogs was 0.08 (range, 0.05 to 0.69). There was a significant difference between mean UPCR before and after castration. CONCLUSIONS AND CLINICAL RELEVANCE: In this study, pathological proteinuria was not detected in sexually intact male dogs. Positive results for a urine dipstick test should be interpreted with caution in sexually intact male dogs and should be confirmed by assessment of the UPCR. An increased UPCR in sexually intact male dogs may be considered abnormal.

Bilateral renal T-cell lymphoma with hepatic infiltration and secondary polycythemia in a dog: Utility of cytology slides.

This is a case of bilateral renal T-cell lymphoma associated with secondary erythrocytosis in a dog. This case is distinctive in using clonality combined with immunocytochemistry to support the diagnosis, thus emphasizing the utility of cytology slides when histology is unavailable. This combination may be a unique canine lymphoma entity.

Lymphome à cellules T rénal bilatéral avec infiltration hépatique et polycythémie secondaire chez un chien : utilité des lames de cytologie. Nous présentons un cas de lymphome à cellules T rénal bilatéral associé à une érythrocytose secondaire chez un chien. Ce cas est distinct parce qu'il fait appel à la clonalité combinée à l'immunocytochimie pour appuyer le diagnostic, ce qui souligne l'utilité des lames de cytologie lorsqu'une histologie n'est pas disponible. Cette combinaison pourrait être une entité unique de lymphome canin.(Traduit par Isabelle Vallières).

Rivaroxaban demonstrates in vitro anticoagulant effects in canine plasma.

Rivaroxaban is an oral direct factor X inhibitor used in human thrombotic disorders and its oral administration makes it an attractive potential anticoagulant for dogs. The objective of this study was to evaluate the in vitro anticoagulant effect of rivaroxaban on canine pooled platelet-poor plasma (PPP). Pooled PPP was collected from 20 healthy adult Beagle dogs. Aliquots of pooled citrated PPP were treated in vitro with DMSO solutions of rivaroxaban (98% purity) to obtain 19 final concentrations ranging from 0 to 1000 mg/L of drug. Samples were immediately submitted for the following coagulation assays: prothrombin time (PT), activated partial thromboplastin time (aPTT), tissue factor-induced thrombin generation and anti-factor Xa activity. Concentration-effect data were analyzed with various nonlinear regression models for stimulatory or inhibitory effects. Rivaroxaban caused a concentration-dependent prolongation of all coagulation parameters. Rivaroxaban concentration for 50% baseline inhibition of the propagation phase of thrombin (rate index) was 0.024 mg/L, and for 50% baseline inhibition of the optical density in the anti-factor Xa activity assay was 0.053 mg/L. At these concentrations, PT and aPTT remained within the reference range. Two-fold prolongation from baseline of PT and aPTT was achieved with higher concentrations, i.e. 1.24 and 1.69 mg/L, respectively. Thrombin generation was completely suppressed by concentrations ≥0.8 mg/L. In conclusion, rivaroxaban showed an in vitro concentration-dependent anticoagulant effect on canine plasma. Thrombin generation and anti-factor Xa activity were more sensitive and accurate than PT and aPTT in detecting the anticoagulant effect of rivaroxaban.