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).

Evaluation of B-lines with 2 point-of-care lung ultrasound protocols in cats with radiographically normal lungs.

OBJECTIVE: To compare 2 point-of-care lung ultrasound (LUS) protocols for quantification of B-lines in cats without evidence of respiratory disease based on history, physical examination, and thoracic radiography. DESIGN: Prospective observational study. SETTING: Single center, veterinary teaching hospital. ANIMALS: Fifty-seven cats without respiratory disease based on history, physical examination, and thoracic radiographs. INTERVENTIONS: All cats had 2 point-of-care LUS protocols performed bilaterally: a regional protocol (veterinary bedside lung ultrasound evaluation [VetBLUE]) and a more comprehensive vertical sweeping (VS) protocol. The total number of B-lines per cat, number of sites with B-lines, and maximal number of B-lines at each site were recorded and compared. MEASUREMENTS AND MAIN RESULTS: Ten cats (18%) had at least 1 B-line identified with VetBLUE, versus 29 (51%) with VS. Comparing protocols, VS had a statistically higher total number of B-lines per cat, higher number of sites with B-lines, and higher maximal number of B-lines per site. B-lines that were too numerous to count were identified at a single location in 1 cat with VetBLUE and 2 cats with VS. A maximum of 3 B-lines were identified at all other positive sites regardless of the protocol used. On average, it took 1.79 times longer to complete VS bilaterally compared to VetBLUE (median [interquartile range]: 140 [33] and 78 [14] s, respectively) (P = 0.001). CONCLUSIONS: This study demonstrates it is not uncommon to identify a single or even multiple B-lines in 1 or several sites on LUS in cats deemed to be clinically free of respiratory pathology-essential knowledge when using LUS as a screening test and to monitor intrathoracic lesions. In cats asymptomatic for respiratory disease, VS generally identifies more B-lines than VetBLUE, likely because it assesses a larger lung surface area. The sonographic identification of B-lines should be interpreted considering the LUS protocol used, history, and other diagnostics to determine their clinical significance.

Evaluation of the host cytokine response in dogs with sepsis and noninfectious systemic inflammatory response syndrome.

OBJECTIVE: To quantify plasma cytokine concentrations in dogs with sepsis and noninfectious systemic inflammation and to evaluate the association between plasma cytokines and outcome in dogs with sepsis. DESIGN: Prospective, observational cohort study. SETTING: University teaching hospital. ANIMALS: Forty-five dogs with sepsis, 10 dogs with noninfectious systemic inflammation (nSIRS), and 15 healthy controls were consecutively enrolled from June 2015 to February 2016 and followed to hospital discharge. Dogs with sepsis satisfied ≥2 SIRS criteria and had a documented or highly suspected bacterial infection. Dogs with nSIRS satisfied ≥2 SIRS criteria but had no evidence of infection. Dogs <3 kg and those with documented coagulopathy were excluded. INTERVENTIONS: Measurement of inflammatory cytokines and high-mobility group box-1 (HMGB-1) was performed on each group. MEASUREMENTS AND MAIN RESULTS: High-mobility group box-1 concentrations were analyzed by ELISA. Plasma concentrations of 13 cytokines were measured in singlet using multiplex magnetic bead assays. Kruskal-Wallis with Dunn's multiple comparison tests were used to compare biomarker concentrations between groups. Mann-Whitney U-tests were used to compare biomarker concentrations between survivors and nonsurvivors. Associations between biomarkers were evaluated using Spearman's correlation coefficients. Independent outcome predictors were identified using multivariable logistic regression. Alpha was set at 0.05. Concentrations of interleukin (IL)-6, C-X-C motif chemokine (CXCL)-8, keratinocyte-derived chemokine (KC)-like, C-C motif chemokine ligand 2 (CCL2), and HMGB-1 were significantly greater in dogs with sepsis versus healthy controls (all P ≤ 0.034). In dogs with sepsis, only CCL2 was independently associated with survival (odds ratio [OR] 0.996, 95% CI 0.993-0.999, P = 0.004). A cut-off of 385 pg/mL for CCL2 was 80% sensitive and 91.4% specific for nonsurvival (area under the ROC curve [AUROC] 0.866). CONCLUSIONS: Dogs with sepsis have significantly increased concentrations of HMGB-1 and inflammatory cytokines, including IL-6, CXCL8, and KC-like. Increased CCL2 concentration is a negative prognostic indicator in dogs with sepsis. These findings should be confirmed using duplicate analyses in larger, distinct populations of dogs with sepsis before applying them to clinical patients.

Determining prognosis in canine sepsis by bedside measurement of cell-free DNA and nucleosomes.

OBJECTIVE: To investigate the use of plasma cell-free DNA (cfDNA) and nucleosome concentrations as prognostic biomarkers in canine sepsis. DESIGN: Prospective, observational cohort study conducted from June 2015 to February 2016. SETTING: University teaching hospital. ANIMALS: Forty-five dogs with sepsis, 10 dogs with nonseptic systemic inflammatory response syndrome (nSIRS), and 15 healthy controls were consecutively enrolled and followed to hospital discharge. Patients were eligible for enrollment if they met ≥2 SIRS criteria and had a documented or highly suspected bacterial infection. Dogs <3 kg or with a known coagulopathy were excluded. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Acute Patient Physiology and Laboratory Evaluation scores (APPLE) were calculated and outcomes recorded. Plasma cfDNA was measured using a benchtop fluorimeter. Plasma nucleosome concentrations were determined by ELISA. Plasma nucleosome and cfDNA concentrations in dogs with sepsis or nSIRS were compared to those of healthy controls and cfDNA concentrations in septic dogs with and without bacteremia were compared. Associations between cfDNA concentrations and nucleosomes, leukocyte count, neutrophil count, and APPLE scores were evaluated. For septic dogs, cfDNA concentrations relative to neutrophil count and nucleosome concentrations in survivors and nonsurvivors were compared. Alpha was set at 0.05. cfDNA concentrations were significantly higher in dogs with sepsis or nSIRS compared to healthy controls (P < 0.0001 and P = 0.0034, respectively). Nucleosome concentrations were significantly higher in dogs with sepsis compared to healthy controls (P = 0.007). There was limited association between cfDNA and nucleosome concentrations (rs  = 0.266), and no association between cfDNA concentration and leukocyte count, neutrophil count, and APPLEfull scores. Concentrations of cfDNA were positively correlated with APPLEfast score (rs  = 0.335, P = 0.025); however, cfDNA concentrations were significantly higher in dogs with bacteremia (P = 0.0299). In dogs with sepsis, cfDNA concentrations relative to neutrophil count were significantly higher in nonsurvivors than in survivors (P = 0.008). CONCLUSIONS: In dogs with sepsis, high cfDNA concentrations relative to neutrophil count are associated with nonsurvival. Point-of-care cfDNA measurement may aid identification of bacteremia.

Concentrations of Plasma Nucleosomes but Not Cell-Free DNA Are Prognostic in Dogs Following Trauma.

Trauma is common in dogs and causes significant morbidity and mortality, but it remains a challenge to assess prognosis in these patients. This study aimed to investigate the use of plasma cell-free DNA (cfDNA) and nucleosome concentrations as prognostic biomarkers in canine trauma. Using a prospective, observational case-control study design, 49 dogs with trauma were consecutively enrolled from 07/2015 to 10/2017 and followed to hospital discharge. Dogs with animal trauma triage (ATT) scores ≥3 at presentation were eligible for enrollment. Dogs <3 kg or with pre-existing coagulopathies were excluded. Thirty-three healthy control dogs were also enrolled. Illness and injury severity scores were calculated using at-presentation data. Plasma cfDNA was measured in triplicate using a benchtop fluorimeter. Plasma nucleosome concentrations were determined in duplicate by ELISA. Mann-Whitney U tests were used to compare biomarker concentrations between groups and between survivors and non-survivors. Associations between biomarkers were evaluated using Spearman's correlation coefficients. Alpha was set at 0.05. Concentrations of cfDNA and nucleosomes were significantly higher in injured dogs compared to healthy controls (P ≤ 0.0001). Nucleosomes and cfDNA concentrations were positively correlated (rs 0.475, P < 0.001). Concentrations of both cfDNA and nucleosomes were correlated with shock index (rs 0.367, P = 0.010, rs 0.358, P = 0.012 respectively), but only nucleosomes were correlated with ATT (rs 0.327, P = 0.022) and acute patient physiology and laboratory evaluation (APPLE) scores (rs 0.356, P = 0.012). Median nucleosome concentrations were significantly higher in non-survivors than in survivors [8.2 AU (3.1-26.4) vs. 1.6 AU (0.5-5.2); P = 0.01]. Among illness severity scores, only APPLE was discriminant for survival (AUROC 0.912, P < 0.001). In summary, in moderately-severely injured dogs, high nucleosome concentrations are significantly associated with non-survival.

High Mobility Group Box-1 and Pro-inflammatory Cytokines Are Increased in Dogs After Trauma but Do Not Predict Survival.

Trauma is common in dogs and causes significant morbidity and mortality, but it remains challenging to predict the prognosis of dogs with traumatic injuries. This study aimed to quantify plasma high-mobility group box-1 (HMGB-1) and cytokine concentrations in dogs with moderate-to-severe trauma, and to evaluate the association between these biomarkers and the injury severity and survival to discharge. Using a prospective, observational case-control study design, 49 dogs with an animal trauma triage (ATT) score ≥3 were consecutively enrolled from 07/2015 to 10/2017 and followed to hospital discharge. Dogs <3 kg and those with pre-existing coagulopathies were excluded. Thirty three healthy control dogs were also enrolled. Illness and injury severity scores including the acute patient physiologic and laboratory evaluation (APPLE) were calculated using at-presentation data. Plasma HMGB-1 concentrations were measured by ELISA; concentrations of 13 cytokines were measured using multiplex bead-based assays and separately concentrations of 4 cytokines were measured using a multiplex canine-specific ELISA. All biomarkers were measured in duplicate. Mann-Whitney U tests were used to compare biomarker concentrations between groups and between survivors and non-survivors. Associations between biomarkers were evaluated using Spearman's correlation coefficients. Independent predictors of survival were identified using multivariable logistic regression. Alpha was set at 0.05. Plasma concentrations of HMGB-1, interleukin-6, C-X-C motif chemokine-8, keratinocyte chemoattractant-like, and C-C chemokine ligand-2 were significantly greater in injured dogs vs. controls (all P ≤ 0.011). In univariate analyses, HMGB-1 was significantly greater in non-survivors 46.67 ng/mL (8.94-84.73) compared to survivors 6.03 ng/mL (3.30-15.75), (P = 0.003). Neither HMGB-1 or the cytokines were associated with survival independent of illness severity as measured by the APPLE score, however.

Serum myoglobin, creatine kinase, and cell-free DNA in endurance sled dogs and sled dogs with clinical rhabdomyolysis.

OBJECTIVE: To evaluate whether cell-free DNA (cfDNA) concentrations are increased in dogs with exertional rhabdomyolysis and whether concentrations are correlated with serum myoglobin concentration and creatine kinase activity. DESIGN: Observational cohort study. SETTING: Yukon Quest 1,000-mile International Sled Dog Race 2015. ANIMALS: Twelve normal competitive sled dogs; 5 dogs with rhabdomyolysis. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Blood was collected from all confirmed cases of exertional rhabdomyolysis and compared to the winning team at the midrace point. Results indicate that median cfDNA did not increase, but decreased by the race finish (prerace = 314.2 ng/mL versus midrace = 283.7 ng/mL versus postrace = 249.5 ng/mL). There were no rises in median cfDNA in dogs with rhabdomyolysis (255 ng/mL) negating its potential utility as a measure of acute skeletal muscle compromise. In contrast, myoglobin concentration and creatine kinase activity at the midrace point for normal dogs were significantly lower than dogs with rhabdomyolysis. Values for myoglobin and creatine kinase were strongly positively correlated (R = 0.91). CONCLUSIONS: cfDNA is not a useful biomarker for exertional rhabdomyolysis in contrast to myoglobin and creatine kinase. Further evaluation of timing and clinical signs suggests that exertional rhabdomyolysis occurs early in endurance activities. Among the dogs with rhabdomyolysis, the dog that demonstrated clinical signs had the highest serum creatine kinase activity and myoglobin concentration.

Measurement of plasma cell-free DNA concentrations in dogs with sepsis, trauma, and neoplasia.

OBJECTIVES: To determine if cell-free DNA (cfDNA) was identifiable in canine plasma, to evaluate 3 techniques for the measurement of plasma cfDNA concentrations in dogs presented to an emergency service, and to compare the plasma cfDNA concentrations of healthy dogs to those with sepsis, trauma, and neoplasia. DESIGN: Retrospective study of banked canine plasma samples collected between May 2014 and December 2014. SETTING: Dogs presented to the emergency service of a university veterinary teaching hospital. ANIMALS: Plasma cfDNA was measured on residual plasma samples obtained from 15 dogs with sepsis, 15 dogs with moderate-severe trauma, 15 dogs diagnosed with a sarcoma. Plasma cfDNA was also measured in 15 healthy dogs. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Assay linearity, repeatability, and reproducibility were evaluated. Quantification of cfDNA was performed in duplicate on diluted citrated plasma and following DNA purification using 2 fluorescence assays (SYBR-Gold; Quant-iT) and by ultraviolet absorbance spectroscopy. Fluorescence intensities (FIs) were converted to cfDNA concentrations using standard curves. Median FI values and cfDNA concentrations were compared to healthy controls using the Kruskal-Wallis test, with adjustment for multiple comparisons. Alpha was set at 0.05. Both assays had excellent linearity, and acceptable repeatability and reproducibility. Compared to controls, plasma cfDNA concentrations were significantly increased in dogs with sepsis or moderate-severe trauma with both assays (P ≤ 0.003). Dogs with neoplasia had significantly increased cfDNA concentrations with the Quant-iT assay only (P = 0.003). When measurements were performed on purified DNA, only dogs with moderate-severe trauma had significantly increased cfDNA concentrations (P < 0.001; SYBR-Gold assay). CONCLUSIONS: cfDNA can be readily identified in canine plasma using 2 fluorescence assays. DNA extraction offers no advantage over direct measurement. Compared to healthy controls, dogs with sepsis or moderate-severe trauma have significantly increased plasma cfDNA concentrations.

Cranial vena cava syndrome secondary to cryptococcal mediastinal granuloma in a cat.

The successful management of cranial vena cava syndrome with suspected secondary chylothorax due to mediastinal cryptococcal granuloma in a 4-year-old male domestic shorthair cat is described. Treatment included long-term antifungal medication, short-term corticosteroids, intermittent thoracocentesis, rutin, octreotide, and enalapril.

Syndrome de la veine cave crâniale secondaire à un granulome médiastinal àCryptococcuschez un chat. La gestion réussie d'un syndrome de veine cave crâniale accompagnée d'un chylothorax secondaire suspecté causé par un granulome médiastinal à Cryptococcus chez un chat commun mâle âgé de 4 ans est décrite. Le traitement a inclus une médication antifongique à long terme, des corticostéroïdes à court terme, une thoracentèse intermittente, de la rutine, de l'octréotide et de l'énalapril.(Traduit par Isabelle Vallières).

A clinical review of the pathophysiology, diagnosis, and treatment of pyothorax in dogs and cats.

OBJECTIVE: To review the current literature in reference to the pathophysiology, diagnosis, and treatment of pyothorax in dogs and cats. ETIOLOGY: Pyothorax, also known as thoracic empyema, is characterized by the accumulation of septic purulent fluid within the pleural space. While the actual route of pleural infection often remains unknown, the oral cavity and upper respiratory tract appear to be the most common source of microorganisms causing pyothorax in dogs and cats. In human medicine, pyothorax is a common clinical entity associated with bacterial pneumonia and progressive parapneumonic effusion. DIAGNOSIS: Thoracic imaging can be used to support a diagnosis of pleural effusion, but cytologic examination or bacterial culture of pleural fluid are necessary for a definitive diagnosis of pyothorax. THERAPY: The approach to treatment for pyothorax varies greatly in both human and veterinary medicine and remains controversial. Treatment of pyothorax has classically been divided into medical or surgical therapy and may include administration of antimicrobials, intermittent or continuous thoracic drainage, thoracic lavage, intrapleural fibrinolytic therapy, video-assisted thoracic surgery, and traditional thoracostomy. Despite all of the available options, the optimal treatment to ensure successful short- and long-term outcome, including the avoidance of recurrence, remains unknown. PROGNOSIS: The prognosis for canine and feline pyothorax is variable but can be good with appropriate treatment. A review of the current veterinary literature revealed an overall reported survival rate of 83% in dogs and 62% in cats. As the clinical presentation of pyothorax in small animals is often delayed and nonspecific, rapid diagnosis and treatment are required to ensure successful outcome.