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.

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.

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.

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.

Marked hyperphosphatasemia associated with an acute leukemia in a Great Dane.

This is the report of a 5-year-old male neutered Great Dane with an extreme leukocytosis (544.9 × 10(9) cells/L; RI 5.2-13.9 × 10(9) cells/L) characterized by highly atypical round cells. Cellular morphologic features such as cytoplasmic membrane blebs, a high nuclear-to-cytoplasmic ratio, and nuclear indentations and irregularities and large nucleoli, as well as immunocytochemistry for CD3 and CD79, myeloperoxidase cytochemistry, and clonality testing were not conclusive for myeloid or lymphoid origin. Marked alkaline hyperphosphatasemia was present at the first visit (2783.0 U/L; RI 6-80.0 U/L), followed by a 5-fold increase (14,000 U/L) a week later, identified as being mostly contributed by the bone-ALP isoform (11,062 U/L; RI 0-30 U/L). In addition, the atypical leukocytes were strongly positive for cytoplasmic ALP activity. In vitro lysis of a heparin blood sample resulted in a 1.7-fold increase of ALP activity, supporting the origin of the hyperphosphatasemia at least in part from the leukemic cell population. To the authors' knowledge, this is a unique case of alkaline hyperphosphatasemia, due at least to a leukemic cell population producing a bone-ALP isoform, regardless of the exact nature of the leukemia.

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.

Bronchoalveolar lavage and pulmonary histopathology in harp seals (Phoca groenlandica) experimentally infected with Otostrongylus circumlitus.

The objective of this study was to characterize pathologic changes associated with experimental infection of harp seals (Phoca groenlandica) with the lungworm Otostrongylus circumlitus (Metastrongyloidea: Crenosomatidae). The leukocyte differential cell count in samples obtained by unguided bronchoalveolar lavage (BAL) and the intensity of the histologic lesions in the lungs were assessed in seven harp seals experimentally exposed to 300 infective, third-stage O. circumlitus larvae. Seven unexposed harp seals were used as controls. First-stage larvae were observed in the feces of three of the seven exposed seals at 38, 42, and 45 days postexposure (dpe). Adult nematodes were found in the right primary bronchi of two of these three seals at necropsy 53 dpe. Fifty-six BALs were performed on the 14 seals. No statistical difference was observed between the exposed and control seals and among the four sampling times in percentage of neutrophils and macrophages in the BAL fluid. A significant difference was observed between the exposed and control seal groups in the percentage of eosinophils (P<0.0001), the count of eosinophils having increased by a factor of 70.4 in exposed seals. Significant statistical differences were observed between exposed and control seals in intensity of interstitial inflammation (P=0.001), bronchitis (P=0.02), bronchiolitis (P=0.04), alveolitis (P=0.03), and interstitial granulomatous inflammation (P=0.04). Our findings showed that harp seals are susceptible to infection with O. circumlitus. However, parasitic infections were transient and of low intensity, at least under our experimental conditions.

Can sonographic findings predict the results of liver aspirates in dogs with suspected liver disease?

While abdominal ultrasound and ultrasound-guided fine-needle aspiration cytology are often combined to help determine the type of liver disease in dogs, little is known about the relationship that may exist between the results of these tests. We hypothesized that specific sonographic findings, or combinations of findings, may predict results of liver ultrasound-guided fine-needle aspiration cytology. Hepatic and extrahepatic sonographic findings were recorded prospectively using a standardized form in 70 dogs with clinically suspected liver disease and in which liver ultrasound-guided fine-needle aspiration cytology was performed. The predictive value of sonographic findings in regard to the category of cytology results was assessed with stepwise logistic regression analysis. Sonographic detection of a hepatic mass (> or = 3cm; risk ratio [RR] 3.83, 95% Wald confidence intervals [95% CI] 2.42-3.93, P = 0.0036), ascites (RR 3.82, 95% CI 1.94-4.28, P = 0.0044), abnormal hepatic lymph node(s) (RR 3.01, 95% CI 1.22-4.88, P= 0.0262), and abnormal spleen (RR 3.26, 95% CI 1.20-3.85, P = 0.0274) were the most predictive of liver neoplasia on cytology. Conversely, sonographic detection of hepatic nodules (< 3cm; RR 1.97, 95% CI 0.95-2.96, P = 0.0666) was most predictive of vacuolar hepatopathy on cytology. In dogs with suspected liver disease, several sonographic findings, alone or combined, are thus predictive of liver ultrasound-guided fine-needle aspiration cytology results. In the light of the fact that ultrasound-guided fine-needle aspiration cytology of the liver has limitations, these predictabilities could influence the selection of diagnostic tests to reach a reliable diagnosis.

What is your diagnosis? Coelomic mass in a rock dove (Columba livia).

A 15-year-old domestic rock dove (Columbia livia) was presented to the Service de Médecine Zoologique of the Université de Montréal with a 10-day history of lethargy and chronic weight loss of 1-year duration. Ultrasonographic evaluation of the coelomic cavity revealed the presence of effusion and a well-defined mass containing variably sized hypoechoic nodules of unknown origin. Cytologic evaluation of fine-needle aspirates of the mass indicated a malignant epithelial tumor consisting of round cells arranged individually and in clusters, with large nuclei, prominent nucleoli, moderate anisocytosis and anisokaryosis, atypical mitoses, occasional binucleation and multinucleation, and large numbers of ruptured cells. A diagnosis of seminoma was established on histologic sections obtained at necropsy. The cytologic features of this tumor were similar to those of canine seminomas except for the presence of lymphocytes, which were not observed in significant numbers in cytologic smears or histologic sections. Seminoma should be included in the differential diagnosis for a space-occupying mass causing organ displacement within the coelomic cavity of a male bird.

Transgenic mice expressing the p75 CCAAT-displacement protein/Cut homeobox isoform develop a myeloproliferative disease-like myeloid leukemia.

The p75 CCAAT-displacement protein/Cut homeobox (CDP/Cux) isoform was previously reported to be overexpressed in human breast cancers. To investigate its oncogenic potential, we engineered two transgenic mouse lines expressing p75 CDP/Cux under the control of the mouse mammary tumor virus-long terminal repeat. The FVB strain of mouse is generally used in the generation of mouse models for breast cancer. The transgene was introduced into the hprt locus of 129/Ola embryonic stem cells and, following germ line passage, was backcrossed onto the FVB and C57BL/6 mouse strains. Here, we describe the phenotype of p75 CDP/Cux transgenic virgin female mice of the first backcross generations. We report that after a long latency period, approximately 33% of mice from two independent transgenic lines and from backcrosses into either the FVB or the C57BL/6 strains succumbed to a similar disease characterized by splenomegaly, hepatomegaly, and frequent infiltration of leukocytes into nonhematopoietic organs like the kidneys and lungs. Although an excess of B or T cells was observed in three diseased mice, in 17 other cases, histologic and flow cytometry analyses revealed the expansion of a population of neutrophils in the blood, spleen, and bone marrow. The increase in neutrophils correlated with signs of anemia and thrombocytopenia, whereas there was no indication of a reactive process. Therefore, p75 CDP/Cux transgenic mice displayed heightened susceptibility to a disease defined as a myeloproliferative disease-like myeloid leukemia. These results indicate that the overexpression of p75 CDP/Cux could alter homeostasis in the hematopoietic compartment.