Protothecosis and Toxoplasma gondii co-infection in a dog from Nova Scotia, Canada.

A 2-year-old, neutered male mixed-breed dog from Nova Scotia, Canada was evaluated for panuveitis, polyuria, polydipsia, and hind limb ataxia. Toxoplasmosis was diagnosed ante-mortem based on markedly increased Toxoplasma gondii titers. The post-mortem examination confirmed systemic toxoplasmosis and demonstrated disseminated protothecosis. This article documents the first reported case of canine protothecosis in Atlantic Canada. Key clinical message: This case report demonstrates that protothecosis should be a clinical consideration for dogs in Canada. Co-infection with other organisms may occur, which may mask clinical signs and potentially delay diagnosis.

Protothécose et co-infection à Toxoplasma gondii chez un chien de la Nouvelle-Écosse, Canada. Un chien de race mixte mâle castré de 2 ans de la Nouvelle-Écosse, au Canada, a été évalué pour une panuvéite, une polyurie, une polydipsie et une ataxie des membres postérieurs. La toxoplasmose a été diagnostiquée antemortem sur la base d'une augmentation marquée des titres de Toxoplasma gondii. L'autopsie a confirmé la toxoplasmose systémique et mis en évidence une protothécose disséminée. Cet article documente le premier cas signalé de protothécose canine en Atlantique. Message clinique clé: Ce rapport de cas démontre que la protothécose devrait être une considération clinique pour les chiens au Canada. Une co-infection avec d'autres organismes peut survenir, ce qui peut masquer les signes cliniques et potentiellement retarder le diagnostic.(Traduit par Dr Serge Messier).

Identification of single-nucleotide variants associated with susceptibility to Salmonella in pigs using a genome-wide association approach.

BACKGROUND: Salmonella enterica serovars are a major cause of foodborne illness and have a substantial impact on global human health. In Canada, Salmonella is commonly found on swine farms and the increasing concern about drug use and antimicrobial resistance associated with Salmonella has promoted research into alternative control methods, including selecting for pig genotypes associated with resistance to Salmonella. The objective of this study was to identify single-nucleotide variants in the pig genome associated with Salmonella susceptibility using a genome-wide association approach. Repeated blood and fecal samples were collected from 809 pigs in 14 groups on farms and tonsils and lymph nodes were collected at slaughter. Sera were analyzed for Salmonella IgG antibodies by ELISA and feces and tissues were cultured for Salmonella. Pig DNA was genotyped using a custom 54 K single-nucleotide variant oligo array and logistic mixed-models used to identify SNVs associated with IgG seropositivity, shedding, and tissue colonization. RESULTS: Variants in/near PTPRJ (p = 0.0000066), ST6GALNAC3 (p = 0.0000099), and DCDC2C (n = 3, p < 0.0000086) were associated with susceptibility to Salmonella, while variants near AKAP12 (n = 3, p < 0.0000358) and in RALGAPA2 (p = 0.0000760) may be associated with susceptibility. CONCLUSIONS: Further study of the variants and genes identified may improve our understanding of neutrophil recruitment, intracellular killing of bacteria, and/or susceptibility to Salmonella and may help future efforts to reduce Salmonella on-farm through genetic approaches.

Dual RNA-Seq characterization of host and pathogen gene expression in liver cells infected with Crimean-Congo Hemorrhagic Fever Virus.

Crimean-Congo hemorrhagic fever virus (CCHFV) is a tick-borne virus that can cause a hemorrhagic fever in humans, with a case fatality rate of up to 40%. Cases of CCHFV have been reported in Africa, Asia, and southern Europe; and recently, due to the expanding range of its vector, autochthonous cases have been reported in Spain. Although it was discovered over 70 years ago, our understanding of the pathogenesis of this virus remains limited. We used RNA-Seq in two human liver cell lines (HepG2 and Huh7) infected with CCHFV (strain IbAr10200), to examine kinetic changes in host expression and viral replication simultaneously at 1 and 3 days post infection. Through this, numerous host pathways were identified that were modulated by the virus including: antiviral response and endothelial cell leakage. Notably, the genes encoding DDX60, a cytosolic component of the RIG-I signalling pathway and OAS2 were both shown to be dysregulated. Interestingly, PTPRR was induced in Huh7 cells but not HepG2 cells. This has been associated with the TLR9 signalling cascade, and polymorphisms in TLR9 have been associated with poor outcomes in patients. Additionally, we performed whole-genome sequencing on CCHFV to assess viral diversity over time, and its relationship to the host response. As a result, we have demonstrated that through next-generation mRNA deep-sequencing it is possible to not only examine mRNA gene expression, but also to examine viral quasispecies and typing of the infecting strain. This demonstrates a proof-of-principle that CCHFV specimens can be analyzed to identify both the virus and host biomarkers that may have implications for prognosis.

Identification of genetic variation in equine collagenous lectins using targeted resequencing.

Collagenous lectins are a family of soluble pattern recognition receptors that play an important role in innate immune resistance to infectious disease. Through recognition of carbohydrate motifs on the surface of pathogens, some collagenous lectins can activate the lectin pathway of complement, providing an effective means of host defense. Genetic polymorphisms in collagenous lectins have been shown in several species to predispose animals to a variety of infectious diseases. Infectious diseases are an important cause of morbidity in horses, however little is known regarding the role of equine collagenous lectins. Using a high-throughput, targeted re-sequencing approach, the relationship between genetic variation in equine collagenous lectin genes and susceptibility to disease was investigated. DNA was isolated from tissues obtained from horses submitted for post-mortem examination. Animals were divided into two populations, those with infectious or autoinflammatory diseases (n = 37) and those without (n = 52), and then subdivided by dominant pathological process for a total of 21 pools, each containing 4-5 horses. DNA was extracted from each horse and pooled in equimolar amounts, and the exons, introns, upstream (approximately 50 kb) and downstream (approximately 3 kb) regulatory regions for the 11 equine collagenous lectin genes and related MASP genes were targeted for re-sequencing. A custom target capture kit was used to prepare a sequencing library, which was sequenced on an Illumina MiSeq. After implementing quality control filters, 4559 variants were identified. Of these, 92 were present in the coding regions (43 missense, 1 nonsense, and 48 synonymous), 1414 in introns, 3029 in the upstream region, and 240 in the downstream region. In silico analysis of the missense short nucleotide variants identified 12 mutations with potential to disrupt collagenous lectin protein structure or function, 280 mutations located within predicted transcription factor binding sites, and 95 mutations located within predicted microRNA binding elements. Analysis of allelic association identified 113 mutations that segregated between the infectious/autoinflammatory and non-infectious populations. The variants discovered in this experiment represent potential genetic contributors to disease susceptibility of horses, and will serve as candidates for further population-level genotyping. This study contributes to the growing body of evidence that pooled, high-throughput sequencing is a viable strategy for cost-effective variant discovery.

Pancreatitis and Systemic Coronavirus Infection in a Ferret (Mustela putorius furo).

A 1-y-old spayed female ferret (Mustela putorius furo) was referred for additional diagnostic evaluation after physical examination by the referring veterinarian revealed a cranial abdominal mass. The ferret had a 2-wk history of inappetence, weight loss, and lethargy. On presentation, the ferret was thin, and an approximately 3-cm mass was palpable in the cranial abdomen. No other abnormalities were noted. Abdominal ultrasonography confirmed the presence of a soft-tissue structure, with a moderate blood supply and mesenteric lymphadenopathy. Fine-needle aspirates of the mass were nondiagnostic. Exploratory laparotomy revealed multiple nodules and thickened tissues throughout the mesentery, a thickened and nodular pancreas, and a small amount of free abdominal fluid. Histopathology of mesenteric, lymphatic, and pancreatic biopsies revealed suppurative pancreatitis and necrotizing and pyogranulomatous mesenteric steatitis. Positive immunohistochemistry for feline coronavirus confirmed a diagnosis of ferret systemic coronavirus disease (FSCD). The ferret was treated medically with oral prednisolone, improved dramatically, and was still doing well 22 mo after diagnosis. Although FSCD has been reported extensively, this case is noteworthy for the presence of suppurative pancreatitis and the positive long-term outcome after corticosteroid therapy.

Interferon-gamma-dependent inhibition of late allergic airway responses and eosinophilia by CD8+ gammadelta T cells.

We have previously shown that CD8(+)gammadelta T cells decrease late allergic airway responses, airway eosinophilia, T helper 2 cytokine expression and increase interferon-gamma (IFN-gamma) expression. We hypothesized that the effects of CD8(+)gammadelta T cells were IFN-gamma mediated. Brown Norway rats were sensitized to ovalbumin on day 1. Cervical lymph node CD8(+)gammadelta T cells from sensitized animals were treated with antisense oligodeoxynucleotide (5 micromol/l) to inhibit IFN-gamma synthesis or control oligodeoxynucleotide and 3.5 x 10(4) CD8(+)gammadelta T cells were injected intraperitoneally into sensitized recipients on day 13. Rats were challenged with aerosolized ovalbumin on day 15 and lung resistance was monitored over an 8 hr period, after which bronchoalveolar lavage was performed. Control oligodeoxynucleotide treated gammadelta T cells decreased late airway responses and eosinophilia in bronchoalveolar lavage. There was a complete recovery of late airway responses and a partial recovery of airway eosinophilia in recipients of antisense oligodeoxynucleotide treated cells. Macrophage ingestion of eosinophils was frequent in rats administered gammadeltaT cells but reduced in recipients of antisense oligodeoxynucleotide treated cells. These results indicate that CD8(+)gammadelta T cells inhibit late airway responses and airway eosinophilia through the secretion of IFN-gamma. Defective or altered gammadelta T-cell function may account for some forms of allergic asthma.

Medio de telurito para aislar y tipear C. diphtheriae

traducido y resumido por la Oficina Sanit. Panam. del Pub. Health Rep. sbre. 10, 1948