Reference Intervals for Blood Analytes of Adult Aquarium-Housed Russian Sturgeon Acipenser gueldenstaedtii.

Russian Sturgeon Acipenser gueldenstaedtii are an important, critically endangered, roe-producing species. Despite a wealth of knowledge pertaining to other members of family Acipenseridae, there is very limited published information regarding baseline blood analytes in Russian Sturgeon. The objectives of this study were (1) to establish reference intervals for a suite of hematological and biochemical data and (2) to compare plasma chemistry data to two point-of-care (POC) cartridges, tested on the VetScan iSTAT 1 analyzer, that use heparinized whole blood for the assessment of clinically normal, aquacultured adult Russian Sturgeon sedated with eugenol (AQUI-S 20E) at a single institution. Reference intervals are reported. The calculated hematocrit measured by the POC analyzer tended 4-5% lower than the spun packed cell volume, confirming the importance of spun packed cell volume as a reliable measurement of red blood cell mass. Various analytes, notably whole-blood urea nitrogen, glucose, sodium, total carbon dioxide, chloride, ionized calcium, and anion gap, were significantly different by both POC cartridges. This study successfully produced reference intervals for blood analytes in adult Russian Sturgeon under managed care and creates a foundation for future studies into the effects of extrinsic and intrinsic factors and variations of analytical methodologies on blood analytes in this species.

Molecular diagnosis using RNAscope in-situ hybridization in canine malignancies.

Immunohistochemistry has been used extensively to evaluate protein expression in clinical and research settings. However, immunohistochemistry is not always successful in veterinary medicine due to the lack of reliable antibody options, poor tissue preservation, labor-intensive staining, and antigen-retrieval optimization processes. RNAscope in-situ hybridization (ISH) is a powerful technology that uses a specific sequence probe to identify targeted mRNA. In this study, we demonstrate RNAscope ISH in 4 common canine malignancies, which are traditionally diagnosed by histopathology and immunohistochemistry. Probes were designed for commonly targeted mRNA markers of neoplastic tumors; these included c-kit in mast cell tumor, microphthalmia-associated transcription factor in malignant melanoma, ionized calcium-binding adapter molecule-1 in histiocytic sarcoma, and alkaline phosphatase in osteosarcoma. A strong staining signal was obtained by these 4 targets in each canine malignancy. These results support the use of RNAscope ISH for definitive diagnosis in canine malignancies.

L'immunohistochimie a grandement été utilisée pour évaluer l'expression de protéines dans des environnements clinique et de recherche. Toutefois, l'immunohistochimie n'est pas toujours un gage de réussite en médecine vétérinaire étant donné le manque d'options pour des anticorps fiables, la mauvaise conservation des tissus, la demande intensive de travail pour la coloration et les processus d'optimisation de recouvrement des antigènes. L'utilisation de l'hybridation in situ (ISH) et du RNAscope est une technologie puissante qui utilise une sonde avec une séquence spécifique afin d'identifier l'ARNm ciblé. Dans la présente étude, nous démontrons l'utilisation de l'ISH RNAscope pour quatre tumeurs canines fréquentes, qui sont traditionnellement diagnostiquées par histopathologie et immunohistochimie. Les sondes furent élaborées pour des marqueurs ARNm fréquemment ciblés de tumeurs néoplasiques; ceux-ci incluaient c-kit pour les tumeurs à cellules mastocytaires, le facteur associé à la transcription de la microphtalmie lors de mélanome malin, la molécule-1 adaptative à l'attachement du calcium ionisé lors de sarcome histiocytaire et la phosphatase alcaline lors d'ostéosarcome. Un signal fort de coloration fut obtenu par ces quatre cibles dans chacun des types de tumeurs. Ces résultats supportent l'utilisation d'ISH RNAscope pour le diagnostic définitif lors de tumeurs canines.(Traduit par Docteur Serge Messier).

Characterization of a novel picornavirus isolated from moribund aquacultured clownfish.

Over the last decade, a number of USA aquaculture facilities have experienced periodic mortality events of unknown aetiology in their clownfish (Amphiprion ocellaris). Clinical signs of affected individuals included lethargy, altered body coloration, reduced body condition, tachypnea, and abnormal positioning in the water column. Samples from outbreaks were processed for routine parasitological, bacteriological, and virological diagnostic testing, but no consistent parasitic or bacterial infections were observed. Histopathological evaluation revealed individual cell necrosis and mononuclear cell inflammation in the branchial cavity, pharynx, oesophagus and/or stomach of four examined clownfish, and large basophilic inclusions within the pharyngeal mucosal epithelium of one fish. Homogenates from pooled external and internal tissues from these outbreaks were inoculated onto striped snakehead (SSN-1) cells for virus isolation and cytopathic effects were observed, resulting in monolayer lysis in the initial inoculation and upon repassage. Transmission electron microscopy of infected SSN-1 cells revealed small round particles (mean diameter=20.0-21.7 nm) within the cytoplasm, consistent with the ultrastructure of a picornavirus. Full-genome sequencing of the purified virus revealed a novel picornavirus most closely related to the bluegill picornavirus and other members of the genus Limnipivirus. Additionally, pairwise protein alignments between the clownfish picornavirus (CFPV) and other known members of the genus Limnipivirus yielded results in accordance with the current International Committee on Taxonomy of Viruses criteria for members of the same genus. Thus, CFPV represents a proposed new limnipivirus species. Future experimental challenge studies are needed to determine the role of CFPV in disease.

The effect of diet on the gastrointestinal microbiome of juvenile rehabilitating green turtles (Chelonia mydas).

Threatened and endangered green turtles (Chelonia mydas) are unique because as juveniles they recruit from pelagic to near-shore waters and shift from an omnivorous to primarily herbivorous diet (i.e. seagrass and algae). Nevertheless, when injured and ill animals are admitted to rehabilitation, animal protein (e.g. seafood) is often offered to combat poor appetite and emaciation. We examined how the fecal microbiome of juvenile green turtles changed in response to a dietary shift during rehabilitation. We collected fecal samples from January 2014 -January 2016 from turtles (N = 17) in rehabilitation at the Georgia Sea Turtle Center and used next generation sequencing to analyze bacterial community composition. Samples were collected at admission, mid-rehabilitation, and recovery, which entailed a shift from a mixed seafood-vegetable diet at admission to a primarily herbivorous diet at recovery. The dominant phyla changed over time, from primarily Firmicutes (55.0%) with less Bacteroidetes (11.4%) at admission, to primarily Bacteroidetes (38.4%) and less Firmicutes (31.8%) at recovery. While the microbiome likely shifts with the changing health status of individuals, this consistent inversion of Bacteroidetes and Firmicutes among individuals likely reflects the increased need for protein digestion, for which Bacteroidetes are important. Firmicutes are significant in metabolizing plant polysaccharides; thus, fewer Firmicutes may result in underutilization of wild diet items in released individuals. This study demonstrates the importance of transitioning rehabilitating green turtles to an herbivorous diet as soon as possible to afford them the best probability of survival.

In vitro effects of the chemotherapy agent water-soluble micellar paclitaxel (Paccal Vet) on canine hemangiosarcoma cell lines.

Paccal Vet (Osamia Pharmaceuticals) is a water-soluble nanoparticle micellar formulation of the drug paclitaxel that is well tolerated in dogs. This study evaluated the in vitro effect of Paccal Vet on two canine hemangiosarcoma (HSA) cell lines and their expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF). Paccal Vet caused HSA cell death in a dose- and time-dependent manner. The 50% inhibitory concentration (IC50 ) for the two HSA cell lines were 7 to 610 ng/mL, which are clinically achievable. Cell cycle analysis through flow cytometry showed cell cycle arrest at G2/M phase. Annexin-V and caspase 3/7 activity assays showed significant increases in apoptosis in correlation with the IC50 in each cell line. Reverse transcriptase-PCR was performed on the cell lines to validate the gene expression of VEGF and bFGF. Results obtained from this study support future studies involving the use of paclitaxel (micellar) for treatment of canine HSA.

Molecular epidemiology of Epizootic haematopoietic necrosis virus (EHNV).

Low genetic diversity of Epizootic haematopoietic necrosis virus (EHNV) was determined for the complete genome of 16 isolates spanning the natural range of hosts, geography and time since the first outbreaks of disease. Genomes ranged from 125,591-127,487 nucleotides with 97.47% pairwise identity and 106-109 genes. All isolates shared 101 core genes with 121 potential genes predicted within the pan-genome of this collection. There was high conservation within 90,181 nucleotides of the core genes with isolates separated by average genetic distance of 3.43 × 10-4 substitutions per site. Evolutionary analysis of the core genome strongly supported historical epidemiological evidence of iatrogenic spread of EHNV to naïve hosts and establishment of endemic status in discrete ecological niches. There was no evidence of structural genome reorganization, however, the complement of non-core genes and variation in repeat elements enabled fine scale molecular epidemiological investigation of this unpredictable pathogen of fish.