Macrocyclic lactone toxicity due to abamectin in farm dogs without the ABCB1 gene mutation

Abstract

CASE HISTORY: A 5-year-old entire female Huntaway from a sheep and beef farm was one of four dogs that developed clinical signs including hypersalivation, depression, blindness and ataxia after the death of another dog. A 4-year-old female Huntaway farm dog from a second farm was observed to be sitting down more often than usual on the day after being fed part of a calf carcass that had been treated with an abamectin pour-on.

CLINICAL FINDINGS: The first dog was ataxic and depressed but did respond to sound. The second dog presented with an acute onset of blindness, mydriasis, absence of a menace response, hypersalivation, gait abnormalities (e.g. high stepping gait and ataxia), and depression. Other presenting signs included muscle tremors, dehydration and difficulty eating. No abnormalities were detected from routine haematology and biochemistry. Analysis of samples of plasma from both dogs revealed concentrations of abamectin of 0.149 mg/L and 0.260 mg/L for the first and second dogs, respectively. Buccal swabs taken from the first dog for DNA testing for the ABCB1 gene mutation, gave a negative result.

DIAGNOSIS: In addition to the presenting signs which suggested a toxicosis, both dogs had measurable concentrations of abamectin in plasma confirming exposure.

CLINICAL RELEVANCE: Farm dogs exposed to concentrated pour-on products containing abamectin have been poisoned and recover or die. The product labels do not carry any warnings as to the risk of poisoning to dogs. This paper discusses two incidents affecting six farm dogs, but the authors are aware of more toxicoses in farm dogs exposed to abamectin.     

Outcome of intensity‐modulated radiation therapy‐based stereotactic radiation therapy for treatment of canine nasal carcinomas

Stereotactic radiation therapy (SRT) has emerged as a convenient definitive treatment modality in veterinary medicine, but few studies exist evaluating outcome with treatment for canine nasal tumors, and no studies report the treatment of one single tumor histotype. This retrospective, observational study evaluates toxicity, response, and survival in 17 dogs with nasal carcinomas treated with SRT. Dogs received a median of 3000 centigray in three fractions via 6‐MV linear accelerator. Eighty‐eight percent of patients (n = 15) demonstrated clinical benefit. Of dogs with repeated CT imaging (n = 10), 60% (n = 6) achieved a partial response and 10% (n = 1) achieved a complete response. Median progression‐free survival (PFS) was 359 days. Median survival time (MST) was 563 days. Among dogs evaluable for acute toxicity, 50% (n = 10) developed low grade toxicity (grade 1, n = 4; grade 2, n = 1). No patients developed grade 3 toxicity. 16 dogs (87%) evaluable over the long term developed signs consistent with possible late toxicity. The majority of late toxicities were mild (alopecia, hyperpigmentation, and leukotrichia n = 10; ocular discharge and keratoconjunctivitis sicca n = 5). Thirty‐seven percent of patients (n = 6) developed seven possible grade 3 late toxicities (blindness, n = 3; fistula, n = 1; seizures, n = 3), which were difficult to distinguish from progressive disease in most patients. Of the prognostic factors evaluated (demographics, tumor stage, dosimetric data, epistaxis, facial deformity, clinical response, image‐based response, nonsteroidal anti‐inflammatory drugs, and chemotherapy), only clinical response was a positive prognostic factor on MST (P < .00). No factors were found to be significantly associated with PFS.     

Influence of type of starch and feeding management on glycaemic control in diabetic dogs

The present study evaluated the effects of two diets with different starch sources and two feeding methods on the glycaemic control in dogs with diabetes mellitus. The diets had similar nutrient contents (40% starch and 16% dietary fibre), one formulated with 46% of broken rice and the other with 42% sorghum and 10% lentils (as-fed). Ten client-owned diabetic dogs were fed with each diet for 2 months, in a crossover design. Five dogs received NPH human insulin and food every 12 h (feeding method 1), and the other five received insulin every 12 h but were fed three times a day (feeding method 2). In feeding method 2, morning insulin was higher than the evening dose and dogs received the second meal after 4 to 5 h of the morning insulin and meal. Parameters evaluated included insulin dosage, 12- and 8-h glycaemic curves, complete blood count, biochemical profile and urinalysis. Glycaemic curves were analysed by ANOVA with repeated measures. Glycaemic control parameters (fasting, mean, minimum and maximum glycaemia and serum fructosamine) and glucose area under the curve (AUC) were calculated and analysed by paired t test (p < 0.05). In feeding method 1, dogs fed the sorghum-based diet presented lower mean (p = 0.04) and minimum blood glucose concentrations (p = 0.03), and a tendency to lower maximum blood glucose (p = 0.06) and glucose AUC (p = 0.08) than when fed the rice-based diet. When food was provided twice a day, the ingestion of the rice-based diet resulted in higher post-prandial glucose response than the diet with sorghum and lentil. In feeding method 2, there was no effect of diet on the assessed parameters (p > 0.05). No differences in insulin dosage were observed between groups or feeding methods (p > 0.05). Providing two meals a day followed by insulin administration associated with the sorghum- and lentil-based diet improved glycaemic control in diabetic dogs.     

A topical aqueous calcineurin inhibitor for the treatment of naturally occurring keratoconjunctivitis sicca in dogs

Purpose The purpose of this study was to evaluate the efficacy of an aqueous calcineurin inhibitor, SCY‐641, in the treatment of naturally occurring canine immune‐mediated keratoconjunctivitis sicca (KCS). Methods A randomized, double‐masked, placebo‐controlled clinical study of 56‐day duration was performed in dogs with naturally occurring immune‐mediated KCS assigned to treatment with either topical twice‐daily aqueous calcineurin inhibitor solution (SCY‐641) or artificial tears (placebo) by the study administrator. Clinical examination and Schirmer tear tests (STT) were performed prior to therapy and at days 7, 14, 28, and 56 after initiation of treatment. Results Twenty dogs were enrolled in the study with ten receiving placebo and 10 receiving SCY‐641 in one or both eyes. No adverse effects were noted with any treatment. There were no significant differences in mean STT values in dogs in group either at day 0 (prior to therapy) or after 7 days of treatment. At 14, 28, and 56 days after initiation of treatment, mean STT and increase in STT over baseline in dogs treated with SCY‐641 were significantly higher than in dogs treated with placebo (P < 0.04). Conclusions SCY‐641 was well tolerated by dogs with naturally occurring KCS, and by 14 days after initiating therapy, dogs treated with SCY‐641 had significantly higher STT than placebo‐treated dogs. These preliminary results indicate that topical SCY‐641, in a stable clear aqueous solution, is efficacious in a spontaneous model of KCS and warrants further evaluation as a treatment of immune‐mediated KCS.     

Evaluation of the long-acting somatostatin analogue Octreotide in the management of insulinoma in three dogs

The response of dogs with insulinoma to surgical and medical management is variable, with the majority developing intractable hypoglycaemia. A long-acting somatostatin analogue, Octreotide (SMS 201–995; Sandostatin) has been useful in the management of hypoglycaemia in humans with insulinoma, and preliminary reports suggest a beneficial clinical response in dogs with insulinoma. The present study objectively evaluated Octreotide in the management of three dogs with immunohistochemically confirmed insulinoma. Octreotide had no benefit over placebo, and little effect on circulating glucose and insulin concentrations despite clearly detectable plasma concentrations of Octreotide. No clinical improvement was apparent in two dogs given Octreotide over a period of two and three weeks. These results contrast with the positive clinical responses noted previously and indicate that further placebo controlled, objective studies are necessary before clear statements on the treatment of insulinoma with Octreotide are made.     

Acute postretinal blindness: ophthalmologic,neurologic, and magnetic resonance imaging findings in dogs and cats (seven cases)

Objective To describe the ophthalmologic, neurologic, and magnetic resonance imaging (MRI) findings of seven animals with acute postretinal blindness as sole neurologic deficit. Methods Medical records were reviewed to identify dogs and cats with postretinal blindness of acute presentation, that had a cranial MRI performed as part of the diagnostic workup. Only animals lacking other neurologic signs at presentation were included. Complete physical, ophthalmic, and neurologic examinations, routine laboratory evaluations, thoracic radiographs, abdominal ultrasound, electroretinography, and brain MRI were performed in all animals. Cerebrospinal fluid analysis and postmortem histopathologic results were recorded when available. Results Four dogs and three cats met the inclusion criteria. Lesions affecting the visual pathways were observed on magnetic resonance (MR) images in six cases. Location, extension, and MRI features were described. Neuroanatomic localization included: olfactory region with involvement of the optic chiasm (n = 4), pituitary fossa with involvement of the optic chiasm and optic tracts (n = 1), and optic nerves (n = 1). Of all lesions detected, five were consistent with intracranial tumors (two meningiomas, one pituitary tumor, two nasal tumors with intracranial extension), and one with bilateral optic neuritis that was confirmed by cerebrospinal fluid analysis. Histologic diagnosis was obtained in four cases and included one meningioma, one pituitary carcinoma, one nasal osteosarcoma, and one nasal carcinoma. Conclusions Central nervous system (CNS) disease should be considered in dogs and cats with acute blindness, even when other neurologic deficits are absent. This study emphasizes the relevance of MRI as a diagnostic tool for detection and characterization of CNS lesions affecting the visual pathways.     

In vitro and in vivo Effects of Lufenuron on Dermatophytes Isolated from Cases of Canine and Feline Dermatophytoses*

Lufenuron is a benzyl‐urea phenol compound that inhibits chitin synthesis and is used as an insecticide. Its efficacy in the therapy of dermatophytosis in dogs and cats was evaluated in several clinical studies, with contradictory results. We assessed the in vitro susceptibility of dermatophytes isolated from dogs and cats to lufenuron, and the clinical response of skin lesions to the drug. Dermatophyte cultures isolated from clinical cases were exposed to lufenuron by three different methods: direct application and application of whole blood or subcutaneous tissue samples obtained from a lufenuron‐treated healthy dog. No inhibition of dermatophyte growth was observed in any of the samples after 1 week of incubation. Eight dogs and six cats with skin lesions were included in the in vivo survey. Results indicated that six of seven skin lesions that were diagnosed as being caused by dermatophytes did not respond to lufenuron whereas six of seven skin lesions that were not caused by dermatophytes improved. We concluded that lufenuron, in the way it was administered in this study, had no inhibitory activity on dermatophytes in vitro or in vivo and its clinical use as an anti‐fungal agent is questionable. An immunomodulatory effect of the drug is, however, possible.     

Effects of a 1% hydrocortisone conditioner on haematological and biochemical parameters, adrenal function testing and cutaneous reactivity to histamine in normal and pruritic dogs

The purpose of this double-blinded study was to examine the effects of a 1% hydrocortisone, leave-on conditioner on haematological and biochemical parameters, adrenal function tests, and cutaneous reaction to serial dilutions of histamine phosphate in normal dogs and those with pruritic dermatitis. Groups 1 and 2 each consisted of eight normal dogs. Seven pruritic dogs comprised Group 3. All dogs were bathed twice weekly for 6 weeks. Groups 1 and 3 had 1% hydrocortisone conditioner applied after each bath. Group 2 had vehicle from the conditioner applied after each bath. The amount of 1% hydrocortisone applied to the treated dogs ranged from 278 to 416 mg/m². Haematological and biochemical analysis and adrenocorticotrophic hormone (ACTH) stimulation tests were performed on all dogs on days 0, 14, 28, and 42. Mean values for all blood and serum parameters remained within normal limits during the study. Post-ACTH cortisol levels were significantly lower in Group 3 compared to Groups 1 and 2 on day 42 (P < 0.05) and when averaged over all days of the study (P < 0.05 and P < 0.01, respectively). Serum alkaline phosphatase levels were significantly lower in Group 3 on day 0, compared to days 14 (P < 0.05), 28 (P < 0.01), and 42 (P = 0.05). All dogs received intradermal injections of buffered saline and five serial dilutions of histamine phosphate on days 0, 14, 28, and 42. No significant differences were apparent among the groups in subjective and objective evaluation of intradermally injected dilutions of histamine. In this study, the use of a 1% hydrocortisone leave-on conditioner did not result in clinically evident adverse effects, and only minor changes in blood parameters were detected. Although mean values in all groups remained within reference ranges throughout the study, the finding of statistically significant lower post-ACTH cortisol concentrations in the pruritic dogs (Group 3) suggests that absorption of hydrocortisone may have occurred. The results of this study also show that this product does not significantly suppress cutaneous reactivity to histamine in normal and pruritic dogs.     

Humoral Antibody Response to Experimental Sarcoptes scabiei var. vulpes Infection in the Dog

Abstract— Humoral antibody responses to experimental Sarcoptes scabiei var. vulpes infection in 10 beagle dogs were demonstrated by an ELISA in three experiments. In the first experiment, four dogs, infected with a relatively high dose of S. scabiei, seroconverted 2 weeks post infection. All four animals showed clinical signs of sarcoptic mange. In the second experiment, six dogs were infected with a relatively low dose of S. scabiei. Three of the dogs developed clinical signs of sarcoptic mange and only in these three were antibodies to S. scabiei demonstrated. Seroconversion occurred 4–5 weeks post infection. The six dogs infected with a low dose of Sarcoptes were re-infected with a relatively low dose of mites 7 weeks after the spontaneous clinical recovery of the three dogs that had exhibited mange following the primary infection. The later three dogs showed a specific antibody response within 1 week and the other three dogs within 2 weeks following the re-infection. Resumen En tres experimentos se demostraron las respuestas humorales de anticuerpos a la infección con Sarcoptes scabiei var. vulpes en 10 perros de la raza Beagle por medio de pruebas ELISA. En el primer experimento cuatro de los perros infectados con una dosis relativamente alta de Sarcoptes, seroconvirtieron 2 semanas despues de la infección. Estos cuatro animales mostraban cuadro clínico de infección. En un segundo experimento seis de los perros se sometieron a una dosis relativamente baja del ácaro. Solo tres de estos perros demostraron cuadro clínico de la infección, y en éstos animales exclusivamente se encontraron anticuerpos de S. scabiei. Cinco o seis semanas después de la infección se produjo la seroconversión de los mismos. Los seis perros infectados con una dosis baja se volvieron a re-infectar con otra dosis relativamente baja siete semanas después de la recuperatión clínica espontánea de los tres perros que habían padecido el cuadro clínico de la sarna, después de la infección previa. Los últimos tres perros produjeron una respuesta de anticuerpos específica en una semana, y los otros tres perros en dos semanas después de la infección.     

Tolerability and pharmacokinetic profile of a novel benzene‐poly‐carboxylic acids complex with cis‐diammineplatinum (II) dichloride in dogs with malignant mammary tumours

The pharmacokinetic profile, tolerability and efficacy of benzene‐poly‐carboxylic acids complex with cis‐diammineplatinum (II) dichloride (BP‐C1) were studied in dogs with mammary cancer. A three‐level response surface pathway designed trial was performed on seven dogs. At each level BP‐C1 was administered subcutaneously daily for 7 days followed by a 7‐day rest period in a dose escalating manner. Adverse events according to VCOG‐CTCAE, performance status and tumour progression were recorded. The pharmacokinetic profile followed a two‐compartment model with rapid absorption, short distribution, and a slow elimination phase. The overall elimination half‐life was 125 h. The maximum tolerated dose of BP‐C1 was estimated to be above 0.46 mg kg^?1. A significant reduction in VCOG‐CTCAE toxicity which correlated negatively with increasing dose was found. The dogs' general performance status remained unchanged. No decrease in total tumour burden was found, although temporary tumour reduction was seen in some target tumours.     

Effect of Leukoreduction on Transfusion‐Induced Inflammation in Dogs

Background: Removal of leukocytes (LR) has been shown to eliminate or attenuate many of the adverse effects of transfusion in experimental animals and humans. Hypothesis/Objectives: Transfusion of stored packed red blood cells (pRBCs) is associated with an inflammatory response in dogs and prestorage LR attenuates the inflammatory response. Animals: Thirteen random‐source, clinically healthy, medium and large breed dogs. Methods: Experimental study. On day 0, animals were examined and baseline blood samples were collected for analysis. Whole blood was then collected for processing with and without LR, and stored as pRBC. Twenty‐one days later, stored pRBCs were transfused back to the donor. Blood samples were collected before and 1 and 3 days after transfusion. Results: In the dogs that received non‐LR pRBCs (n = 6) there was a significant increase from baseline in white blood cell count from a mean (SD) of 8.20 (2.74) to 13.95 (4.60) × 10³ cells/μL (P < .001) and in segmented neutrophil count from a mean (SD) of 5.76 (2.70) to 11.91 (4.71) × 10³ cells/μL (P < .001). There were also significant increases in fibrinogen from a mean (SD) of 129.7 (24.2) to 268.6 (46.7) mg/dL (P < .001) and C‐reactive protein from a mean (SD) of 1.9 (2.1) to 78.3 (39.3) μg/mL (P < .001). There was no significant increase from baseline in any of the markers in the dogs that received LR pRBC (n = 5). Conclusions and Clinical Importance: There is a profound inflammatory response to transfusion in normal dogs, which is eliminated by LR of the pRBC units.     

Long‐term survival with stereotactic radiotherapy for imaging‐diagnosed pituitary tumors in dogs

Published studies on the use of stereotactic radiotherapy for dogs with pituitary tumors are limited. This retrospective observational study describes results of stereotactic radiotherapy for 45 dogs with imaging‐diagnosed pituitary tumors. All dogs were treated at a single hospital during the period of December 2009–2015. The stereotactic radiotherapy was delivered in one 15 Gray (Gy) fraction or in three 8 Gy fractions. At the time of analysis, 41 dogs were deceased. Four were alive and censored from all survival analyses; one dog received 8 Gy every other day and was removed from protocol analyses. The median overall survival from first treatment was 311 days (95% confidence interval 226–410 days [range 1–2134 days]). Thirty‐two dogs received 15 Gy (median overall survival 311 days; 95% confidence interval [range 221–427 days]), and 12 received 24 Gy on three consecutive days (median overall survival 245 days, 95% confidence interval [range 2–626 days]). Twenty‐nine dogs had hyperadrenocorticism (median overall survival 245 days), while 16 had nonfunctional masses (median overall survival 626 days). Clinical improvement was reported in 37/45 cases. Presumptive signs of acute adverse effects within 4 months of stereotactic radiotherapy were noted in 10/45, and most had improvement spontaneously or with steroids. Late effects versus tumor progression were not discernable, but posttreatment blindness (2), hypernatremia (2), and progressive neurological signs (31) were reported. There was no statistical difference in median overall survival for different protocols. Patients with nonfunctional masses had longer median overall survival than those with hyperadrenocorticism (P = 0.0003). Survival outcomes with stereotactic radiotherapy were shorter than those previously reported with definitive radiation, especially for dogs with hyperadrenocorticism.     

Granulocyte function tests in canine infectious diseases: Methods and preliminary clinical results

The in vitro chemotaxis and the intracellular killing capacity of phagocytosed $\text{S. aureus}$ by canine neutrophils were determined. After obtaining values for clinically normal dogs, the neutrophils of thirteen dogs with different kinds of chronic recurrent or persistent infections were tested. No impaired functions of the neutrophils of any of these patients could be demonstrated.     

Gold-induced immune thrombocytopenia in the dog

In two seven-year studies with gold compounds in dogs of both sexes, thrombocytopenia was observed after 45 to 72 months of dosing in three of 14 and two of 14 dogs in high-dose groups that received 2.4 to 3.6 mg/kg of auranofin per day orally or 0.5 to 2.0 mg/kg of gold sodium thiomalate intramuscularly once every three days, respectively. An immune basis for the disorder was suggested by the apparent consumptive nature of the thrombocytopenia (increased bone marrow megakaryocytes and large peripheral blood platelets), the response to corticosteroid therapy and the demonstration of increased platelet-associated immunoglobulin. The latter was demonstrated with a solid phase radioimmunoassay and by electron microscopy using a staphylococcal protein A-colloidal gold conjugate. Platelet-associated immunoglobulin decreased as the platelet counts rose, and in one dog monitored over periods of steroid-induced remissions and subsequent relapses, the amount of platelet-associated immunoglobulin G correlated inversely with the platelet count (r = 0.82). These findings suggest that the long-term administration of gold compounds in dogs is associated with a dose-dependent incidence of thrombocytopenia, which is immune-mediated and similar to that associated with parenteral chrysotherapy in man. The application of tests for platelet-associated immunoglobulin to canine patients with immune thrombocytopenia should be useful in the diagnosis of the disorder in clinical practice.     

Phase II open‐label study of single‐agent hydroxyurea for treatment of mast cell tumours in dogs*

This prospective study evaluated the efficacy and safety of hydroxyurea (HU) in dogs with measurable mast cell tumours (MCTs). Dogs were treated with HU at 60 mg kg^?1per os q24h for 14 days then 30 mg kg^?1 q24h thereafter or until MCT recurrence. Forty‐six dogs were enrolled. The overall response rate was 28%. Two dogs had a complete response (CR) for 256 and 448 days, respectively. Eleven dogs had a partial response for a median duration of 46 days (range, 28–189 days). Grade 2 to 4 neutropenia occurred in eight dogs and grade 4 thrombocytopenia in two. Grade 3–4 anaemia occurred in seven dogs; overall, there was a significant decrease in haematocrit after treatment with HU. The median drop in haematocrit was 10%. This study demonstrated that HU has activity in the treatment of MCTs with mild anaemia being the primary adverse event.     

Efficacy of a pimaricin suspension for treating otitis externa associated with M pachydermatis

A clinical trial was conducted to evaluate the efficacy of a 1 per cent pimaricin suspension in the treatment of canine otitis externa associated with Malassezia pachydermatis. Of 40 dogs examined, M pachydermatis was the sole infection in five ears and was commensal with other organisms in 35 ears. Pimaricin in a 1 per cent suspension was administered twice a day for two weeks. Employing continuous therapy, signs accompanying otitis externa were ameliorated steadily, and satisfactory results were achieved in 33 of the dogs. No noticeable side effects were observed with the pimaricin suspension.     

P‐74 Evaluation of the in vivo effects of Tris‐EDTA and chlorhexidine digluconate 0.15% solution in chronic bacterial otitis externa: 11 cases

The objectives of this study were to evaluate in vivo tolerance, and antimicrobial and clinical activities of a topical otic preparation containing EDTA tromethamine (Tris) and chlorhexidine digluconate 0.15% solution (Otodine^®) in dogs with chronic bacterial otitis externa. Eleven dogs were included. The affected ears were filled with the solution once daily during a 2‐week period. Dogs were evaluated on days 0, 14 and 28. Three clinical parameters (exudate, erythema, pain) and three cytologic parameters (Malassezia, cocci, rods) were scored (0–4 scale) by otoscopic and cytological examinations of otic exudate. Bacterial cultures were performed at each time point. If there were bacteria on cytological examination on day 14, the dogs were treated with the original product, with the addition of enrofloxacin (5%) applied 10 min after the original product, for a further 2 weeks. All 11 cases yielded isolates of resistant gram‐negative bacteria; gram‐positive bacteria were also isolated from six of 11 dogs. On day 14, six of 11 dogs were negative on culture examination; on day 28, 10 of 11 were negative and only one case had a positive culture. On day 14, clinical and microbial scores (cytology) were reduced by 54.6 and 71.1%, respectively, and by 85.7 and 94% on day 28. All cases reported good tolerance of the treatment. The results show that this ear solution was helpful in the management of chronic bacterial otitis externa in dogs and was well tolerated. There seems to be a synergistic effect of the combination of Tris‐EDTA/chlorhexidine digluconate 0.15% solution, and an antimicrobial agent (enrofloxacin) against resistant gram‐positive and gram‐negative bacteria. Funding: Self‐funded.     

Evaluation of fungal flora in normal and diseased canine ears

This study was undertaken to characterize otic fungal flora encountered in normal dogs, atopic dogs with no clinical or cytological evidence of otitis and dogs with otitis externa. Forty‐two normal dogs, 23 atopic dogs and 32 dogs with otitis were included in the study. Samples for otic fungal culture and cytology were obtained from all animals, for a total of 194 ears. Sixty‐seven ear samples (34%) were culture positive for saprophytic fungal organisms, as follows: 43 (64%) Penicillium species, 13 (19%) Aspergillus species and the remaining 17% comprised of various other saprophytic fungal organisms. Cytological evidence of saprophytic fungal colonization or infection was not found in any animal. There was no relationship between positive saprophytic fungal culture and any study group. Thirty‐three ear samples (17%) were positive for Malassezia pachydermatis. Cytological findings of Malassezia were significantly associated with positive culture for Malassezia (P = 0.006 left ear; P = 0.019 right ear). Furthermore, increased numbers of Malassezia led to a higher chance of positive culture (P = 0.003 left ear; P = 0.008 right ear; McNemar’s test). Malassezia pachydermatis was more likely to be cultured from ears with increased cerumen. Ear type (erect or pendulous) was not significantly associated with positive culture for Malassezia or saprophytic fungal organisms. There was no relationship between positive Malassezia culture and any study group; however, Malassezia was more likely to be cultured from individual dogs in the atopic or otitis groups that also had other dermatological signs consistent with allergic dermatitis and/or pyoderma (P = 0.031 left ear; P = 0.005 right ear).     

Establishment of reference ranges and evaluation of in vitro concentration‐dependent platelet inhibition by acetylsalicylic acid for multiple electrode impedance aggregometry in healthy dogs

Acetylsalicylic acid (ASA, aspirin) is an antiplatelet medication used for prevention of thromboembolism. Effects of ASA appear to vary widely between dogs, but the underlying mechanisms are not understood. The Multiplate analyzer is a newer form of whole‐blood impedance aggregometry recently validated for use in healthy dogs. A method utilizing this instrument to measure ASA effects on platelet function has not been established. The goals of this study were to establish reference ranges for the Multiplate in healthy dogs and secondly, to develop a technique to determine the in vitro concentration of ASA needed to cause 50% inhibition of platelet aggregation (IC50). Reference ranges established from 40 dogs at multiple test times for three agonists were consistent with previously published values. In vitro IC50 values were calculated using the sigmoid E_max model in 20 healthy dogs on two occasions to determine individual repeatability. Calculated in vitro IC50 demonstrated four ASA response groups: responder (n = 16), poor responder (n = 1), variable responder (n = 2), and nonresponder (n = 1). Multiplate within‐assay variability was  <10% for area under the curve (AUC), and between‐assay baseline AUC variability was  <15%. The described technique allowed for determination of an in vitro IC50 for ASA in dogs using a multiple electrode impedance aggregometer.     

Occurrence and characterization of gastric Helicobacter spp. in naturally infected dogs

Helicobacter-like organisms are frequently observed in the stomach of dogs but the relationship between these microorganisms and gastric pathology has not been clearly established. Different species of helicobacters are known to be present in the canine stomach but their specific prevalence in naturally infected dogs is unknown. The aims of this study were to isolate and characterize helicobacters in canine gastric biopsies, to compare the commonly used tests for the identification of Helicobacter spp. and to determine the occurrence of these species in dogs. Twenty-three out of 25 dogs (92%) were positive for Helicobacter-like organisms in cytological screening. Culture was successful from biopsies of 5/25 dogs. The isolates were analyzed by electron microscopy, biochemical and physiological tests, whole protein analysis and 16S rDNA sequencing. Helicobacter felis was identified in four samples and Helicobacter bizzozeronii in one sample. Only the whole protein analysis in combination with electron microscopy was able to clearly discriminate the two species. Compared to the high prevalence of Helicobacter-like organisms, the occurrence of H. felis and H. bizzozeronii, was low (17 and 4%, respectively). No Flexispira rappini-like organisms or H. salomonis were detected. Electron microscopy revealed that H. bizzozeronii-like microorganisms were present in three additional biopsies where we were unable to culture any Helicobacter-like organisms. These observations indicate that in the stomach of dogs not all helicobacters are culturable. The unculturable bacteria appeared to be the prevalent ones and may represent different spiral organisms. The presence of distinct helicobacters with different characteristics can reflect different roles in the pathogenesis of canine gastric disease.