Hypersensitivity pneumonitis associated with inhalation of Lycoperdon spores (lycoperdonosis) in a 3-month-old English setter dog in Quebec.

A 3-month-old female English setter dog was presented to the Faculty of Veterinary Medicine of the Université de Montréal (Quebec) with acute respiratory distress. The dog had moderately increased C-reactive protein concentrations, and thoracic radiographs revealed a moderate, caudodorsal, nodular-to-miliary alveolo-interstitial pulmonary pattern that was worse in the perihilar region. Initial differential diagnoses included a fungal pneumonia (e.g., blastomycosis or histoplasmosis). Cytology of the bronchoalveolar lavage revealed several round, green structures ~2 µm in diameter, consistent with fungal spores. The dog was hospitalized, but within 24 h the respiratory condition deteriorated and euthanasia was elected. Post-mortem panfungal PCR and sequencing tests identified the spores as Lycoperdon sp. Retrospectively, the owners recalled that the dog had played in a wood pile with mushrooms and had sneezed in a cloud of spores, implying inhalation of Lycoperdon spores. This is the first report of a confirmed case of canine lycoperdonosis in eastern Canada (Quebec), and the radiographic features in this case differed slightly from previous reports. Diagnosis before bronchoalveolar lavage analysis was challenging, as spore inhalation was not initially reported. Although the disease is infrequently reported in dogs, this case report reminds veterinarians to consider lycoperdonosis as a differential diagnosis when addressing animals presented with acute dyspnea with similar radiographic lesions, and highlights the importance of history and cytology in diagnosing this condition. Key clinical message: Hypersensitivity pneumonitis secondary to inhalation of Lycoperdon spores must be included in differential diagnoses for a dog with acute onset of respiratory signs and a nodular-to-miliary interstitial pulmonary pattern coalescing in patchy perihilar alveolar pulmonary lesions, and should prompt clinicians to question owners regarding inhalation of mushroom spores.Although cytological examination of a bronchoalveolar lavage reveals the presence of fungal spores, panfungal PCR and sequencing tests are needed to pinpoint the species involved.

Pneumopathie d'hypersensibilité associée à l'inhalation de spores de Lycoperdon (lycoperdonose) chez un chien setter anglais de 3 mois au Québec. Une chienne setter anglais âgée de 3 mois a été présentée à la Faculté de médecine vétérinaire de l'Université de Montréal (Québec) avec une détresse respiratoire aiguë. Le chien présentait des concentrations de protéine C-réactive modérément augmentées et les radiographies thoraciques ont révélé un schéma pulmonaire alvéolo-interstitiel modéré, caudodorsal, nodulaire à miliaire, pire dans la région périhilaire. Les diagnostics différentiels initiaux incluaient une pneumonie fongique (par exemple, blastomycose ou histoplasmose). La cytologie du lavage broncho-alvéolaire a révélé plusieurs structures rondes et vertes d'environ 2 µm de diamètre, compatibles avec des spores fongiques. Le chien a été hospitalisé, mais en 24 heures, l'état respiratoire s'est détérioré et l'euthanasie a été décidée. Les tests panfongiques PCR et de séquençage post-mortem ont identifié les spores comme étant Lycoperdon sp. Rétrospectivement, les propriétaires ont mentionné que le chien avait joué dans un tas de bois avec des champignons et avait éternué dans un nuage de spores, ce qui implique une inhalation de spores de Lycoperdon. Il s'agit du premier rapport d'un cas confirmé de lycoperdonose canine dans l'est du Canada (Québec), et les caractéristiques radiographiques de ce cas différaient légèrement des rapports précédents. Le diagnostic avant l'analyse du lavage broncho-alvéolaire était difficile, car l'inhalation de spores n'avait pas été initialement signalée. Bien que la maladie soit rarement rapportée chez les chiens, ce rapport de cas rappelle aux vétérinaires de considérer la lycoperdonose comme un diagnostic différentiel lorsqu'ils traitent des animaux présentant une dyspnée aiguë avec des lésions radiographiques similaires, et souligne l'importance de l'anamnèse et de la cytologie dans le diagnostic de cette affection.Message clinique clé : La pneumopathie d'hypersensibilité secondaire à l'inhalation de spores de Lycoperdon doit être incluse dans les diagnostics différentiels chez un chien présentant un début aigu de signes respiratoires et un schéma pulmonaire interstitiel nodulaire à miliaire fusionnant dans des lésions pulmonaires alvéolaires périhilaires inégales, et devrait inciter les cliniciens à interroger les propriétaires concernant l'inhalation de spores de champignons.Bien que l'examen cytologique d'un lavage broncho-alvéolaire révèle la présence de spores fongiques, des tests panfongiques PCR et de séquençage sont nécessaires pour identifier les espèces impliquées.(Traduit par Dr Serge Messier).

VALIDATION OF ENZYME-LINKED IMMUNOSORBENT ASSAY TECHNIQUES TO MEASURE SERUM DEHYDROEPIANDROSTERONE (DHEA) AND DHEA-S IN NARWHALS (MONODON MONOCEROS).

Narwhals (Monodon monoceros) are increasingly exposed to anthropogenic disturbances that may increase their stress levels with unknown consequences for the overall population dynamics. The validation and measurement of chronic stress biomarkers could contribute toward improved understanding and conservation efforts for this species. Dehydroepiandrosterone (DHEA) and its sulfated metabolite DHEA-S are collectively referred to as DHEA(S). Serum DHEA(S) concentrations combined in ratios with cortisol [cortisol/DHEA(S)] have been shown to be promising indicators of chronic stress in humans, domestic animals, and wildlife. During field tagging in 2017 and 2018 in Baffin Bay, Nunavut, Canada, 14 wild narwhals were sampled at the beginning and end of the capture-tagging procedures. Serum DHEA(S) were measured with commercially available competitive enzyme-linked immunosorbent assays (ELISA) developed for humans. A partial validation of the ELISA assays was performed by the determination of the intra-assay coefficient of variation, confirmation of the DHEA(S) dilutional linearity, and the calculation of the percentage of recovery. Mean values (nanograms per milliliter ± standard error of the mean) of narwhal serum cortisol, DHEA(S), and cortisol/DHEA(S) ratios, at the beginning and at the end of handling, respectively, are reported (cortisol = 30.74 ± 4.87 and 41.83 ± 4.83; DHEA = 1.01 ± 0.52 and 0.99 ± 0.50; DHEA-S = 8.72 ± 1.68 and 7.70 ± 1.02; cortisol/DHEA = 75.43 ± 24.35 and 84.41 ± 11.76, and cortisol/DHEA-S = 4.16 ± 1.07 and 6.14 ± 1.00). Serum cortisol and cortisol/DHEA-S were statistically higher at the end of the capture (P= 0.024 and P= 0.035, respectively). Moreover, serum cortisol at the end of handling was positively correlated to total body length (P = 0.042) and tended to be higher in males (P = 0.086). These assays proved easy to perform, rapid, and suitable for measuring serum DHEA(S) of narwhals and that calculated cortisol/DHEA(S) are potential biomarkers for chronic stress in narwhals and possibly other cetaceans.

Differential role of MyD88 signaling in Streptococcus suis serotype 2-induced systemic and central nervous system diseases.

Streptococcus suis serotype 2 is an important porcine bacterial pathogen and a zoonotic agent responsible for sudden death, septic shock and meningitis, with exacerbated inflammation being a hallmark of the systemic and central nervous system (CNS) infections. However, S. suis serotype 2 strains are genetically and phenotypically heterogeneous, being composed of a multitude of sequence types (STs) whose virulence greatly varies. Yet, most studies have used 'classical' virulent Eurasian ST1 or ST7 strains, even though ST25 and ST28 strains account for most isolates in North America. While recognition of S. suis by innate immune cells has been associated with the myeloid differentiation primary response 88 (MyD88)-dependent Toll-like receptor (TLR) pathway in vitro, particularly surface-associated TLR2, little information is available regarding its role in vivo. This study demonstrates for the first time a differential role of MyD88 signaling in S. suis-induced systemic and CNS diseases, regardless of strain background diversity. The MyD88-dependent pathway is critical for the development of systemic disease via its role in inflammation, which subsequently controls bacterial burden. However, and differently from what has been described in vitro, TLR2 and TLR4 individually do not contribute to systemic disease, suggesting possible compensation in their absence and/or a collaborative role with other MyD88-dependent TLRs. On the other hand, CNS disease does not necessarily require MyD88 signaling and, consequently, neither TLR2 nor TLR4, suggesting a partial implication of other pathways. Finally, regardless of its notable heterogeneity, recognition of S. suis serotype 2 appears to be similar, indicating that recognized components are conserved motifs.

A Disseminated Cryptococcus gattii VGIIa Infection in a Citron-Crested Cockatoo (Cacatua sulphurea citrinocristata) in Québec, Canada.

Cryptococcus gattii infection in mammals and birds has been confined historically to tropical and subtropical regions in Australia, Southeast Asia, Africa, and South America. Since the early 2000s, numerous reports describe the emergence of C. gattii on the Pacific Coast of North America. We report on a C. gattii infection in an 8-year-old male citron-crested cockatoo (Cacatua sulphurea citrinocristata) hatched on the Canadian Pacific Coast and raised in the province of Québec, Canada. The bird developed a slow growing ulcerated, fleshy, crusty, and hemorrhagic mass infiltrating the left lower rhamphotheca. Cryptococcus gattii infection was confirmed by cytologic examination of a fine needle aspirate of the mass, and results of fungal culture and sequencing. The genotype of the strain was determined to be VGIIa sequence type 20, the strongly overrepresented subgroup found on the Canadian Pacific coast. Minimum inhibitory concentrations for multiple antifungal drugs were determined. The bird received fluconazole but died acutely 55 days after initial presentation. Postmortem examination revealed a disseminated infection, with involvement of the beak, lungs, spleen, and brain.

Encapsulated Streptococcus suis impairs optimal neutrophil functions which are not rescued by priming with colony-stimulating factors.

The porcine pathogen and zoonotic agent Streptococcus suis induces an exacerbated inflammation in the infected hosts that leads to sepsis, meningitis, and sudden death. Several virulence factors were described for S. suis of which the capsular polysaccharide (CPS) conceals it from the immune system, and the suilysin exhibits cytotoxic activity. Although neutrophils are recruited rapidly upon S. suis infection, their microbicidal functions appear to be poorly activated against the bacteria. However, during disease, the inflammatory environment could promote neutrophil activation as mediators such as the granulocyte colony-stimulating factor granulocyte (G-CSF) and the granulocyte-macrophages colony-stimulating factor (GM-CSF) prime neutrophils and enhance their responsiveness to bacterial detection. Thus, we hypothesized that CPS and suilysin prevent an efficient activation of neutrophils by S. suis, but that G-CSF and GM-CSF rescue neutrophil activation, leading to S. suis elimination. We evaluated the functions of porcine neutrophils in vitro in response to S. suis and investigated the role of the CPS and suilysin on cell activation using isogenic mutants of the bacteria. We also studied the influence of G-CSF and GM-CSF on neutrophil response to S. suis by priming the cells with recombinant proteins. Our study confirmed that CPS prevents S. suis-induced activation of most neutrophil functions but participates in the release of neutrophil-extracellular traps (NETs). Priming with G-CSF did not influence cell activation, but GM-CSF strongly promote IL-8 release, indicating its involvement in immunomodulation. However, priming did not enhance microbicidal functions. Studying the interaction between S. suis and neutrophils-first responders in host defense-remains fundamental to understand the immunopathogenesis of the infection and to develop therapeutical strategies related to neutrophils' defense against this bacterium.

Effects of soaked hay on lung function and airway inflammation in horses with severe asthma.

BACKGROUND: Reducing inhaled dust particles improves lung function in horses with severe asthma. Soaked hay is commonly used by owners, but its efficacy in improving lung function and inflammation has not been documented. OBJECTIVES: To measure the effects of soaked hay and alfalfa pellets in horses with severe asthma. ANIMALS: Ten adult horses with severe asthma from a research colony. METHODS: Prospective controlled trial. Horses in clinical exacerbation were housed indoors and allocated to be fed either soaked hay (n = 5) or alfalfa pellets (n = 5) for 6 weeks. Soaked hay was immersed for 45 minutes and dried out hay was discarded between meals. Pulmonary function and clinical scores were measured before and after 2, 4, and 6 weeks. Tracheal mucus scores and bronchoalveolar lavages were performed before and after 6 weeks. Lung function was analyzed with a linear mixed model using log-transformed data. RESULTS: Lung resistance decreased from (median (range)) 2.47 (1.54-3.95) to 1.59 (0.52-2.10) cmH2 O/L/s in the pellets group and from 1.89 (1.2-3.54) to 0.61 (0.42-2.08) cmH2 O/L/s in the soaked hay group over the 6-week period for an average difference of 1.06 cmH2 O/L/s for pellets (95% confidence interval [95% CI]: 0.09-2.04, P = .03, not significant after correction) and 1.31 cmH2 O/L/s for soaked hay (95% CI: -0.23 to 2.85, P < .001, significant). CONCLUSION AND CLINICAL IMPORTANCE: Soaked hay can control airway obstruction in horses with severe asthma. The strict protocol for soaking and discarding dried-out hay in this study could however be considered too great of an inconvenience by owners.

Steamed hay and alfalfa pellets for the management of severe equine asthma.

BACKGROUND: Steaming hay significantly reduces respirable particles and provides a palatable alternative to dry hay for horses with asthma, but there are few prospective studies demonstrating the clinical efficacy of steamed hay. OBJECTIVES: To compare the efficacy of alfalfa pellets and steamed hay in improving lung function and inflammation of horses with severe asthma (SEA). STUDY DESIGN: Controlled crossover study. METHODS: Ten horses with SEA were enrolled and nine completed the study. Horses were housed indoors and fed hay. Once in exacerbation, they were fed pellets and steamed hay for 4 weeks, in a crossover design. Weighted clinical scores and lung function were recorded weekly. Bronchoalveolar lavage fluid (BALF) cytology and mucus scores were recorded before and after each diet. RESULTS: Based on linear mixed model (LMM) analysis, weighted clinical scores significantly improved over time (p < 0.001, no diet effect or time-diet interactions). With pellets, weighted clinical scores decreased from (median (interquartile range)) 13 (5.5) to 2 (1.5), while with steamed hay, they decreased from 10 (9.5) to 6 (8.5). With pellets, lung resistance decreased significantly from a baseline of (mean (SD)) 2.62 (0.77) cmH2O/L/s to 1.17 (0.66), 0.79 (0.54), 0.70 (0.20), 0.62 (0.18) on Weeks 1-4, respectively (LMM with post hoc tests, p < 0.001). With steamed hay, the resistance decreased significantly from a baseline of 2.34 (0.93) cmH2O/L/s to 1.38 (0.49) and 1.51 (0.66) on Weeks 1 and 2, respectively (p < 0.03). Neutrophils BALF decreased significantly with both diets (pellets: 40.2 (24.4) to 20.1 (11.0) %; steamed hay 30.9 (20.2) to 25.7 (17.6) %; LMM, p = 0.006). MAIN LIMITATIONS: A small number of horses in a research setting. Dust was not measured in the stalls. CONCLUSIONS: Clinical scores, lung function and BALF inflammation of horses with SEA improved with steamed hay and pellets, but the effect on lung function was more pronounced with pellets.

The added value of One Health surveillance: data from questing ticks can provide an early signal for anaplasmosis outbreaks in animals and humans.

OBJECTIVE: In 2021, a first outbreak of anaplasmosis occurred in animals and humans in southern Québec, with 64% of confirmed human cases located in Bromont municipality. Ixodes scapularis ticks and Peromyscus mouse ear biopsies collected in Bromont from 2019 to 2021 were analyzed for Anaplasma phagocytophilum (Ap) with the objective of determining whether an early environmental signal could have been detected before the outbreak. METHODS: Samples were collected for a concurrent study aiming to reduce Lyme disease risk. Between 2019 and 2021, up to 14 experimental sites were sampled for ticks and capture of small mammals took place on three sites in 2021. Samples were screened for Ap using multiplex real-time PCR, and genetic strains were identified using a single-nucleotide polymorphism assay. RESULTS: Analyses showed an increase of 5.7% in Ap prevalence in ticks (CI95: 1.5-9.9) between 2019 and 2020, i.e., one year before the outbreak. A majority of Ap-positive ticks were infected with the zoonotic strain (68.8%; CI95: 50.0-83.9) during the study period. In 2021, 2 of 59 captured Peromycus mice were positive for Ap, for a prevalence of 3.4% (CI95: 0.4-11.7). CONCLUSION: We conclude that data collected in Bromont could have provided an early signal for an anaplasmosis risk increasing in the targeted region. This is a reminder that integrated surveillance of tick-borne diseases through structured One Health programs, i.e. systematically integrating data from humans, animals and the environment, can provide useful and timely information for better preparedness and response in public health.

RéSUMé: OBJECTIF: En 2021, suivant une éclosion d'anaplasmoses chez les animaux et les humains dans le sud du Québec, des tiques de l'espèce Ixodes scapularis et des biopsies de souris Peromyscus spp. échantillonées à Bromont, la municipalité où 64 % des cas humains confirmés était localisé, ont été testées pour Anaplasma phagocytophilum (Ap) avec pour objectif de déterminer si un signal environnemental précoce d'augmentation du risque aurait pu être détecté avant l'éclosion. MéTHODE: L'échantillonnage a été réalisé dans le cadre d'une étude visant à réduire le risque de maladie de Lyme. De 2019 à 2021, 14 sites expérimentaux ont été échantillonnés pour les tiques. En 2021, trois sites ont été sélectionnés pour la capture des micromammifères. Les échantillons ont été testés pour la présence d'Ap à l'aide d'un PCR multiplex en temps réelle et les lignées génétiques ont été identifiées grâce à un test de polymorphisme mononucléotidique. RéSULTATS: Les analyses ont montré une augmentation de 5,7 % (IC95% : 1,5­9,9) de la prévalence de Ap entre 2019 et 2020, c'est-à-dire un an avant l'éclosion. Cette augmentation est associée à la présence d'une majorité d'Ap de la lignée zoonotique (68,8 %; IC95% : 50,0­83,9) sur l'ensemble de la période étudiée. En 2021, deux Peromycus spp. capturées sur 59 étaient positives pour Ap pour une prévalence de 3,4 % (IC95% : 0,4­11,7). CONCLUSION: Les données environnementales échantillonnées à Bromont auraient pu fournir un signal précoce de l'augmentation du risque d'anaplasmose dans la région. C'est un rappel que la surveillance intégrée des maladies transmises par les tiques inspirée de l'approche Une seule santé, intégrant systématiquement des données humaines, animales et environnementales, peut fournir des informations utiles et opportunes aux autorités de santé publique.

Brachystemma calycinum D. Don Effectively Reduces the Locomotor Disability in Dogs with Naturally Occurring Osteoarthritis: A Randomized Placebo-Controlled Trial.

Objective. The aim of this randomized placebo-controlled trial was to evaluate the beneficial effect of a whole plant extract of Brachystemma calycinum D. Don (BCD) in naturally occurring osteoarthritis (OA) in dogs. Methods. Dogs had stifle/hip OA and poor limb loading based on the peak of the vertically oriented ground reaction force (PVF) measured using a force platform. At baseline, PVF and case-specific outcome measure of disability (CSOM) were recorded. Dogs (16 per group) were then assigned to receive BCD (200 mg/kg/day) or a placebo. The PVF was measured at week (W) 3 and W6. Locomotor activity was recorded throughout the study duration using collar-mounted accelerometer, and CSOM was assessed biweekly by the owner. Results. BCD-treated dogs had higher PVF at W3 and W6 when compared to Baseline (P < 0.001) and at W6 when compared to placebo-treated dogs (P = 0.040). Higher daily duration (P = 0.024) and intensity (P = 0.012) of locomotor activity were observed in BCD-treated dogs compared to baseline. No significant change was observed in either group for CSOM. Conclusions. Treatment with BCD improved the limb impairment and enhanced the locomotor activity in dogs afflicted by naturally-occurring OA. Those preclinical findings provide interesting and new information about the potential of BCD as an OA therapeutic.

Comparative short-term efficacy of Oridermyl(®) auricular ointment and Revolution(®) selamectin spot-on against feline Otodectes cynotis and its associated secondary otitis externa.

The efficacy of Oridermyl(®) (acaricidal/antibiotic/antifungal/anti-inflammatory ointment) and Revolution(®) (selamectin spot-on) was compared in a blinded randomized study on 24 adult cats with otoacariasis. Twelve cats were treated once daily for 10 d with Oridermyl(®) and 12 cats were treated on Day 0 with Revolution(®). Otitis was assessed with cytological counts of mean number of bacteria and yeast on Days 0 and 10, and scored clinically every other day. All auricular secretions were removed for mite count on Day 10. On Day 0, cytological examination confirmed the presence of secondary bacterial (24/24) and fungal (21/24) infections. No live mites were observed otoscopically after Day 4 and in auricular secretions at Day 10 in both groups. On Day 10, secondary infections were resolved for all cats treated with Oridermyl(®) but were present in all cats treated with Revolution(®). Improvement in clinical signs of otitis over time was superior in the Oridermyl(®) group (P < 0.001).

A retrospective study of 29 cases of otitis media/interna in dairy calves.

Epidemiological data, clinical findings, laboratory data, medical imaging, and outcomes were reviewed in 29 dairy calves with otitis media/interna. Age at admission ranged from 1 to 24 wk. The majority of calves were referred during winter. Clinical signs included drooping ear, ptosis, head tilt, abnormal nystagmus, strabismus, dysphagia, regurgitation, stiff neck, opisthotonos, facial hyperesthesia, and purulent aural discharge. Intranasal endoscopic examination of 5 animals revealed nasopharyngeal collapse in 4. Cerebrospinal fluid (CSF) was abnormal in all of 7 cases. Mycoplasma bovis was cultured from all but 1 case with external ear or tympanic bullae samples (n = 12), and Mycoplasma arginini was cultured from the remaining ear sample. Radiographs of the tympanic bullae were performed in 24 calves, tomodensitometry (CT) in 3 calves and ultrasound in 4 calves. According to medical imaging techniques or necropsy, 69% of the cases were classified as chronic. Mean duration of treatment was 23.3 d. The rate of clinical recovery was 75%.Epidemiological data, clinical findings, laboratory data, medical imaging, and outcomes were reviewed in 29 dairy calves with otitis media/interna. Age at admission ranged from 1 to 24 wk. The majority of calves were referred during winter. Clinical signs included drooping ear, ptosis, head tilt, abnormal nystagmus, strabismus, dysphagia, regurgitation, stiff neck, opisthotonos, facial hyperesthesia, and purulent aural discharge. Intranasal endoscopic examination of 5 animals revealed nasopharyngeal collapse in 4. Cerebrospinal fluid (CSF) was abnormal in all of 7 cases. Mycoplasma bovis was cultured from all but 1 case with external ear or tympanic bullae samples (n = 12), and Mycoplasma arginini was cultured from the remaining ear sample. Radiographs of the tympanic bullae were performed in 24 calves, tomodensitometry (CT) in 3 calves and ultrasound in 4 calves. According to medical imaging techniques or necropsy, 69% of the cases were classified as chronic. Mean duration of treatment was 23.3 d. The rate of clinical recovery was 75%.

RésuméÉtude rétrospective de 29 cas d'otite moyenne/interne chez les veaux laitiers. Les données épidémiologiques, les signes cliniques, les résultats de laboratoire et d'imagerie médicale et l'évolution de 29 veaux atteints d'otite moyenne/interne sont présentés. L'âge à la présentation variait de 1 à 24 semaines. La majorité des veaux ont été admis en hiver. Les signes cliniques incluaient une oreille tombante, une ptose de la paupière, une tête penchée, un nystagmus pathologique, un strabisme, de la dysphagie, des régurgitations, une raideur cervicale, de l'opisthotonos, de l'hyperesthésie faciale et une écoulements purulente de l'oreille. L'endoscopie des voies respiratoires supérieures a révélé un collapse du nasopharynx dans 4 cas sur 5. L'analyse du liquide céphalo-rachidien était anormale chez 7 veaux. Mycoplasma bovis a été isolé de tous les cas à partir d'échantillons d'oreille externe ou de bulle tympanique sauf 1 (n = 12) où Mycoplasma arginini a été isolé. La radiographie des bulles tympaniques a été réalisée sur 24 cas, la tomodensitométrie sur 3 cas et l'échographie sur 4 cas. Selon les techniques d'imagerie médicale ou la nécropsie, 69 % des cas étaient chroniques. La durée moyenne du traitement était de 23,3 jours. Le pronostic était de 75 %.(Traduit par les auteurs).

Assessment of thrombin generation in horses using a calibrated automated thrombogram.

BACKGROUND: The amount of thrombin generated reflects the endogenous thrombin potential (ETP), which depends on the balance of pro- and anticoagulant factors. The calibrated automated thrombogram (CAT) allows for the direct measurement of thrombin generation during the clotting process. OBJECTIVES: (1) To describe the results of the CAT assay in horses, (2) to establish intra-assay and intra- and interindividual variation of thrombin generation in healthy horses, and (3) to compare in vitro low-molecular-weight heparin (LMWH) sensitivity between healthy and sick horses. The hypothesis for the last objective is that inhibition of thrombin generation in sick horses requires higher heparin concentrations. METHODS: The plasma of 10 healthy mixed breed horses was used for the determination of normal thrombin generation parameters (lag time, time to peak, peak thrombin concentration, and ETP). Five of the healthy horses were compared with five horses with systemic inflammatory response syndrome (SIRS). In vitro heparin sensitivity was determined using LMWH. RESULTS: The intra-assay variation was small (<5%) for all parameters. Relatively large intra- and interindividual variation were observed in healthy horses. Four of the five sick horses with SIRS had a thrombogram compatible with a hypercoagulable state. The in vitro heparin sensitivity test suggested decreased sensitivity to LMWH in hypercoagulable states. CONCLUSIONS: The CAT assay could detect coagulopathy in horses. In vivo experiments are needed to confirm that it can be used to monitor responses to LMWH therapy.

Interpretation of cerebrospinal fluid analysis from recumbent cows using different thresholds of red blood cell count.

BACKGROUND: Hemodilution of the cerebrospinal fluid (CSF) could confound interpretation of results. Accurately predicting total nucleated cells count (TNCC) and total protein concentration (TPC) attributable to hemodilution is difficult. OBJECTIVE: To determine the effects of hemodilution on TPC and TNCC in bovine CSF. METHODS: Retrospective review of CSF analysis results of downer dairy cows treated at Centre hospitalier universitaire vétérinaire between January 2006 and December 2014. Descriptive statistics were performed using 3 scenarios. RESULTS: Among the 235 samples included, red blood cell (RBC) count (RBCC) ranged from 0 to 869 220 RBC/µL (median = 6.6), TPC ranged from 0.04 to 6.51 g/L (median = 0.27), and TNCC ranged from 0 to 7500 cell/µL (median = 1.1). Among the 157 samples that had <30 RBC/µL (a threshold used in other species), TPC and TNCC varied between 0.13 and 1.06 g/L (median = 0.27) and between 0 and 31.4 cell/µL (median = 0.6), respectively. Eighty-four samples had TPC <0.25 g/L and TNCC ≤4.5 cell/µL. Among those 84 samples, RBCC varied between 0 and 1290 RBC/µL (median = 4.7). In 20 samples, TNCC was 0 with a variation in RBCC between 0 and 840 RBC/µL (median = 3.9). No strong correlations between RBCC and TNCC and TPC were found. CONCLUSIONS: A cutoff around 200 RBC/µL is proposed as clinically meaninful in bovine CSF. Results between 200 and 1290 RBC/µL are equivocal.

Characterization of fungal exposure and dectin-1 expression in healthy horses and horses with severe asthma.

OBJECTIVE: To quantify dectin-1 expression in bronchoalveolar lavage fluid (BALF), create polyclonal antibodies against equine dectin-1 and localize it in tissues, and quantify fungal exposure in pastured and stabled asthmatic and nonasthmatic horses. SAMPLES: BALF samples from 6 controls and 6 horses with severe asthma. Stored lung and nasal wash samples. PROCEDURES: Dectin-1 expression was quantified by quantitative PCR (qPCR). Purified peptide from equine dectin-1 was used to generate polyclonal antibodies and was confirmed with immunological testing. Fungal exposure was quantified in BALF samples by counting fungal-like intracellular particles in phagocytic cells, by qPCR quantification of the "universal" 18S rRNA fungal gene, and by quantifying 36 specific fungi in equine and dust samples using qPCR assays. RESULTS: Equine dectin-1 was localized in tissues and cells, and functional isoforms were upregulated significantly in BALF after stabling. Pastured horses from both groups had low levels of fungi in BALF, and there was a significant increase in some specific fungi, most notably for Eurotium amstelodami, Wallemia sebi, and Aspergillus niger after stabling. However, stabled asthmatic horses had fewer phagocytized particles, less 18S rRNA signal, and fewer specific fungi compared to nonasthmatic horses. CLINICAL RELEVANCE: Stabling increases exposure to fungi, but asthmatic horses had fewer fungi reaching their lower airways, presumably resulting from congestion and narrowing of the airways. Exposure to fungi could contribute to airway inflammation by increasing dectin-1 functional isoforms, and exposure to indoor molds should be avoided.

Treatment of feline otitis externa due to Otodectes cynotis and complicated by secondary bacterial and fungal infections with Oridermyl auricular ointment.

A blinded randomized study was conducted on 24 cats to confirm the presence of bacterial and/or fungal secondary infections associated with otoacariasis and to verify the efficacy of Oridermyl, an acaricidal/antibiotic/antifungal/anti-inflammatory ointment, for treatment of the primary infestation and secondary infections. Sixteen cats were treated once daily for 10 d; 4 cats were not treated and 4 were treated with a placebo ointment. On Days 0 and 10, ears were swabbed for counts of bacteria and yeasts, for bacterial culture and sensitivity, and examined for determination of the degree of clinical otitis. Auricular secretions were removed for mite counts on Day 10, except for 8 treated cats that were done on Day 30. There was a high number of bacteria and yeasts in most cats and Oridermyl treatment significantly decreased those numbers. Staphylococci were the most frequently isolated bacteria. No live ear mites were found in cats treated with Oridermyl or the placebo ointment.

A primitive neuroectodermal tumor with extension into the cranial vault in a dog.

This paper reports the clinical findings, cytology, diagnostic imaging, and necropsy of an unusual case of a peripheral nervous system neoplasm which, subsequent to a 6-month clinical history, extended into the cranial vault. Necropsy and histology confirmed the diagnosis of a peripheral primitive neuroectodermal tumor.

Serum interleukin 17 concentrations in dogs with immune-mediated hemolytic anemia.

BACKGROUND: Increased serum interleukin 17 (IL-17) concentration has been associated with the immunopathogenesis of autoimmune hemolytic anemia in humans. No data are available about IL-17 in immune-mediated hemolytic anemia (IMHA) of dogs. OBJECTIVES: Monitor changes in serum IL-17 concentration during the acute stages of IMHA in dogs, compared with results in healthy dogs, and its relationship with outcome. ANIMALS: Thirty-one client-owned dogs with primary IMHA and 27 healthy dogs. METHODS: Quantification of serum IL-17 concentration using a commercially available ELISA kit at the time of admission (D0), after 48 hours (D2) and after 96 hours (D4) as compared to concentration in healthy dogs. The IMHA dogs were classified as survivors if discharged from hospital, or nonsurvivors for any cause of in-hospital mortality. RESULTS: Mean serum IL-17 concentration was higher in dogs with IMHA on admission compared with healthy dogs (D0), but this difference was not significant (mean, 19.52 pg/mL vs 10.52 pg/mL, respectively, P = .17). Throughout hospitalization, serum IL-17 concentration significantly decreased in survivors. Serum IL-17 concentration at D0 was not different between survivors and nonsurvivors, but surviving dogs had significantly lower serum IL-17 concentration at D2 and D4 (P = .04 and P = .004, respectively) compared with nonsurviving dogs. No correlation was found between serum IL-17 concentration and serum total bilirubin or lactate concentrations or CBC parameters. CONCLUSION AND CLINICAL IMPORTANCE: Serum IL-17 concentration remained significantly higher in nonsurviving IMHA dogs whereas it significantly decreased during hospitalization in survivors, making serum IL-17 concentration a potential biomarker for severity and response to treatment in IMHA.

Proof-of-concept study: Evaluation of plasma and urinary electrolytes as markers of response to L-asparaginase therapy in dogs with high-grade lymphoma.

Response to chemotherapy is one of the most important prognostic factors in dogs with lymphoma. The objective of this feasibility study was to evaluate if clinical responses to a specific cytotoxic agent (L-asparaginase) could be anticipated by measuring analyte concentrations in plasma and urine concentrations of lymphoma-bearing dogs. We hypothesized that potassium and phosphate concentrations in plasma and urine would be higher in dogs that completely responded to therapy. Plasma and urine samples of dogs with lymphoma were obtained before 12 and 24 hours after intramuscular L-asparaginase injections. Peripheral lymph node volumes were evaluated according to the Veterinary Cooperative Oncology Group standardized criteria. Plasma and urine electrolyte, calcium, phosphate, creatinine, urea, total protein, and albumin concentrations were measured, and the fractional excretions of each electrolyte were calculated. Statistical analyses compared complete vs partial responders using a linear regression model. Contrast analyses were also performed to differentiate the mean of each group, with adjustments made with the Benjamini-Hochberg procedure. Fourteen dogs were included, eight with complete responses, and six with partial responses. Plasma phosphate concentrations were significantly higher at 12 hours (P = .0003) and 24 hours (P = .009) after complete responses to therapy. This study demonstrates the potential use of plasma and urine analyte monitoring after chemotherapy induction. Plasma phosphate measurements represent a potential indicator of early responses to L-asparaginase therapy. Larger population studies are warranted to confirm these preliminary results.

Stability of epidermal growth factor, fibronectin, and alpha-2-macroglobulin in canine serum under different storage conditions.

The objective of this study was to assess whether concentrations of epidermal growth factor (EGF), fibronectin, and alpha(α)-2-macroglobulin in canine serum remain stable under different storage conditions. Serum was obtained from 10 adult dogs and stored for 7 d at room temperature (RT) and at 4°C and for 1, 3, and 6 mo at -20°C. Bacterial cultures of serum were carried out after 7 d at 4°C and at RT. For each dog and time point, EGF, fibronectin, and α-2-macroglobulin were quantified in duplicate by enzyme-linked immunosorbent assay (ELISA). Mean concentrations of each factor at each time point were used for statistical analysis. No bacterial growth was observed in any samples. Compared to baseline (232.24 ± 49.47 pg/mL), EGF concentration was significantly lower after 1 wk of storage at 4°C (135.39 ± 27.12 pg/mL, P = 0.006), but not at RT (315.85 ± 79 pg/mL, P = 0.6) or after 1, 3, or 6 mo of storage at -20°C (220.84 ± 41.07 pg/mL, P = 0.7; 220.98 ± 78.26 pg/mL, P = 0.8; 266.06 ± 20.39 pg/mL, P = 0.4, respectively). Compared to baseline, concentrations of fibronectin after 1 wk of storage at 4°C or at RT and 1, 3, or 6 mo of storage at -20°C were not statistically different. Compared to baseline (186.67 ± 45.20 mg/dL), the concentration of α-2-macroglobulin after 1 wk of storage at 4°C was significantly increased (244.61 ± 58.27 mg/dL, P = 0.002), but not at RT (177.09 ± 26.99 mg/dL, P = 0.2). The differences in concentration after 3 and 6 mo of storage at -20°C were significant compared to baseline (243.32 ± 42.64 mg/dL, P = 0.005 and 56.39 ± 21.78 mg/dL, P < 0.0001, respectively), but not after 1 mo of storage at -20°C (136.79 ± 25.61 mg/dL, P = 0.1). One week of storage at RT has little effect on the stability of EGF, fibronectin, and α-2-macroglobulin in canine serum. Measured factors remain stable for 3 mo of storage at -20°C.

L'objectif de la présente étude était d'évaluer si les concentrations du facteur de croissance épidermique (EGF), de la fibronectine, et de l'alpha (α)-2 macroglobuline dans le sérum canin demeuraient stable sous différentes conditions d'entreposage. Du sérum fut obtenu de 10 chiens adultes et entreposé pendant 7 j à la température de la pièce (RT) et à 4 °C et pendant 1, 3, et 6 mo à −20 °C. Des cultures bactériennes du sérum furent effectuées après 7 j à 4 °C et RT. Pour chaque chien et temps de vérification, l'EGF, la fibronectine et l'α-2 macroglobuline furent quantifiés en duplicata par épreuve immuno-enzymatique (ELISA). Les concentrations moyennes de chaque facteur à chaque temps de vérification furent utilisées pour analyse statistique. Aucune croissance bactérienne ne fut observée dans les différents échantillons. Comparativement à la valeur de base (232,24 ± 49,47 pg/mL), la concentration d'EGF était significativement plus basse après 1 sem d'entreposage à 4 °C (135,39 ± 27,12 pg/mL, P = 0,006), mais pas à RT (315,85 ± 79 pg/mL, P = 0,6) ou après 1, 3, ou 6 mo d'entreposage à −20 °C (220,84 ± 41,07 pg/mL, P = 0,7; 220,98 ± 78,26 pg/mL, P = 0,8; 266,06 ± 20,39 pg/mL, P = 0,4, respectivement). Comparativement à la valeur de base, les concentrations de fibronectine après 1 sem d'entreposage à 4 °C ou à RT et après 1, 3, ou 6 mo d'entreposage à −20 °C n'étaient pas statistiquement différentes. Comparativement à la valeur de base (186,67 ± 45,20 mg/dL), la concentration d'α-2 macroglobuline après 1 sem d'entreposage à 4 °C avait augmenté significativement (244,61 ± 58,27 mg/dL, P = 0,002), mais pas à RT (177,09 ± 26,99 mg/dL, P = 0,2). Les différences dans les concentrations après 3 et 6 mois d'entreposage à −20 °C étaient significatives comparativement à la valeur de base (243,32 ± 42,64 mg/dL, P = 0,005 et 56,39 ± 21,78 mg/dL, P < 0,0001, respectivement), mais pas après 1 mo d'entreposage à −20 °C (136,79 ± 25,61 mg/dL, P = 0,1). Une semaine d'entreposage à RT avait peu d'effets sur la stabilité d'EGF, de fibronectine et d'α-2 macroglobuline dans le sérum canin. Les facteurs mesurés sont demeurés stables pour 3 mo lors d'entreposage à −20 °C.(Traduit par Docteur Serge Messier).

Randomised study of the immunomodulatory effects of azithromycin in severely asthmatic horses.

Neutrophilic inflammation is believed to contribute to the airway obstruction and remodelling in equine asthma. Azithromycin, an antibiotic with immunomodulatory properties, reduces pulmonary neutrophilia and hyper-responsiveness in human asthmatics and decreases airway remodelling in rodent models of asthma. It was therefore hypothesised that azithromycin would improve lung function, mucus accumulation and central airway remodelling by decreasing luminal neutrophilia in severe equine asthma. The effects of a 10-day treatment with either azithromycin or ceftiofur, an antimicrobial without immune-modulating activity, were assessed using a blind, randomised, crossover design with six severe asthmatic horses in clinical exacerbation. Lung function, tracheal mucus accumulation, tracheal wash bacteriology, bronchial remodelling, airway neutrophilia and mRNA expression of proinflammatory cytokines (interleukin (IL)-8, IL-17A, IL-1ß, tumour necrosis factor-α) in bronchoalveolar lavage fluid were evaluated. Azithromycin decreased the expression of IL-8 (P=0.03, one-tailed) and IL-1ß (P=0.047, one-tailed) but failed to improve the other variables evaluated. Ceftiofur had no effect on any parameter. The reduction of neutrophilic chemoattractants (IL-8, IL-1ß) justifies further efforts to investigate the effects of a prolonged treatment with macrolides on airway neutrophilia and remodelling. The lack of efficacy of ceftiofur suggests that severe equine asthma should not be treated with antibiotics at first-line therapy.