Technical note: development of a duodenal cannula for sheep.

A single T-shaped duodenal cannula of silicone rubber with a gutter-type small inner flange was developed for sheep. The barrel of the cannula was 24 mm long with an internal diameter of 12 mm. A polyester surgical mesh (100 mm x 100 mm) was connected to the barrel of the cannula as an anchor. Fibrous tissues grew on the polyester mesh anchor and adhered firmly to the serosa of the intestine, thus leaving no gap and, hence, preventing any leakage of intestinal contents from the side of the fistula. The small (24 mm in diameter) and thin (3 mm in thickness) outer flange of the cannula became buried in the wool and prevented any mechanical disturbance of the cannula by the activity of the animal. The elasticity of the silicone rubber prevented distortion of the duodenum around the barrel of the cannula. No erosion of the tissue between the inner flange of the cannula and the mesh was seen in postmortem observations.     

Oral allergy syndrome induced by tomato in a dog with Japanese cedar (Cryptomeria japonica) pollinosis

A dog with Japanese cedar (Cryptomeria japonica, CJ) pollinosis had oral allergy syndrome (OAS) after ingesting fresh tomato. The dog showed specific IgE to both CJ and tomato allergens. As a negative control, twenty dogs without atopic dermatitis that had no exposure to tomato and no specific IgE to CJ allergen were used. They had no specific IgE to tomato allergen. Furthermore, IgE cross-reactivity was observed between CJ and tomato allergens in the dog. We found that OAS induced by tomato exists in the dog and there is a relationship between CJ and tomato allergens.     

Genetic diversity of <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">spinaciae</Emphasis> in Japan based on phylogenetic analyses of rDNA-IGS and <Emphasis Type="Italic">MAT1</Emphasis> sequences

Twenty-eight isolates of Fusarium oxysporum f. sp. spinaciae (FOS; the causal agent of spinach wilt) collected from Japan were assessed for mating type and subjected to phylogenetic analysis. Mating type analysis revealed all isolates to be MAT1-2, suggesting that there is no sexual recombination within the population. Phylogenetic analyses based on nucleotide sequences of the ribosomal DNA intergenic spacer (IGS) and the mating type locus (MAT1) suggested that FOS is polyphyletic. The cluster analysis based on IGS showed four phylogenetic groups (S1–S4) among the isolates. Two distinct lineages, S1 and S3, included FOS isolates both of the vegetative compatibility group (VCG) types, 0330 and 0331, demonstrating that VCG differentiation in FOS may not necessarily reflect the phylogenetic relationships based on IGS and MAT1-2-1.     

Three evolutionary lineages of tomato wilt pathogen, Fusarium oxysporum f. sp. lycopersici, based on sequences of IGS, MAT1, and pg1, are each composed of isolates of a single mating type and a single or closely related vegetative compatibility group

Three evolutionary lineages of the tomato wilt pathogen Fusarium oxysporum f. sp. lycopersici were found among a worldwide sample of isolates based on phylogenetic analysis of the ribosomal DNA intergenic spacer region. Each lineage consisted of isolates mainly belonging to a single or closely related vegetative compatibility group (VCG) and a single mating type (MAT). The first lineage (A1) was composed of isolates VCG 0031 and MAT1-1; the second (A2) included VCG 0030 and/or 0032 and MAT1-1; and the third (A3) included VCG 0033 and MAT1-2. Race 1 and race 2 isolates belonged to the A1 or A2 lineages, and race 3 belonged to A2 or A3 lineages, suggesting that there is no correlation between race and lineage. However, for the isolates from Japan, race 1 (with one exception), race 2, and race 3 isolates belonged to A2, A1, and A3 lineages, respectively. These results suggest that the races could have evolved independently in each lineage; and in Japan the present races were likely to have been introduced independently after they had evolved in other locations.     

Chimeric mice with humanized liver as a model for testing organophosphate and carbamate pesticide exposure

BACKGROUND

Diagnosis of acute intoxication with organophosphate (OP) or carbamate (CM) pesticides in humans is achieved by measuring plasma butyrylcholinesterase (BuChE) activity. However, BuChE activity is not an ideal biomarker in experimental animal models. The aim of this study was to establish an experimental mouse model for evaluating exposure to OP and CM pesticides by monitoring BuChE activity using chimeric mice in which the liver was reconstituted with human hepatocytes.

RESULTS

A single oral administration of acephate (300 mg/kg), chlorpyrifos (10 mg/kg), fenobucarb (300 mg/kg) or molinate (250 mg/kg) in chimeric mice led to inhibition of >95%, > 95%, 28% and 60% of plasma BuChE activity after 7, 0.5, 0.5 and 7 h, respectively. Dose‐dependent decreases in plasma BuChE activity were also observed for acephate and chlorpyrifos. A 5‐day repeated‐dose study with 10 or 30 mg/kg acephate found a constitutive reduction in plasma BuChE activity to 80% and 70% of pre‐dose levels, respectively.

CONCLUSION

Changes in plasma BuChE activity in chimeric mice with humanized liver clearly reflected the exposure levels of OP and CM pesticides. These results suggest that the humanized‐liver mouse model may be suitable for estimating levels of exposure to these pesticides in humans. © 2017 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

Imatinib elicited a favorable response in a dog with a mast cell tumor carrying a c-kit c.1523A>T mutation via suppression of constitutive KIT activation

Target therapy using the tyrosine kinase inhibitor imatinib is one of the new therapeutic approaches for canine mast cell tumors (MCTs). In the present report, we demonstrate a clinical response to imatinib in a dog with MCT carrying a c-kit c.1523A>T mutation. Moreover, the effect of this mutation on the phosphorylation status of KIT and the inhibitory potency of imatinib on the phosphorylation of the mutant KIT were examined in vitro. A dog with a MCT tumor mass on the right forelimb sole with lymph node metastasis and mastocytemia was treated with imatinib. The MCT mass markedly shrank and mastocytemia became undetectable with 2 weeks of treatment. The lymph node enlarged by metastasis became normal in size with 5 weeks of treatment. From the sequencing analysis of c-kit in tumor cells, a substitution mutation c.1523A>T that alters the amino acid composition (p.Asn508Ile) within the extracellular domain of KIT was identified. The mutant KIT expressed on 293 cells showed ligand-independent phosphorylation and imatinib suppressed this phosphorylation in a dose-dependent manner. From these findings, imatinib was considered to elicit a clinical response in a canine case of MCT via inhibition of the constitutively activated KIT caused by a c-kit c.1523A>T mutation.     

Morphometric and histochemical study of involution in the rat uterus after parturition

Morphological changes to and collagen loss from the rat uterus during postpartum involution were investigated. The expression patterns of collagen type III and myeloperoxidase (MPO) were also determined. Morphological changes were studied on days 1, 3, 5, 10, 15, 20, 22 and 25 postpartum. As a control, diestrus rats’ uterine were used. Specimens from the uterine horn were embedded in paraffin, cut into 8 µm coronal sections, and stained with hematoxylin‐eosin. The thickness of the longitudinal and circular smooth muscle layers and of the endometrium were measured. The collagen content was determined using hydroxyproline analysis. Immunostaining was used to examine the expression of collagen type III on days 1, 3, 5, 10, 15 and 20 postpartum; and MPO on days 1, 3, 5, 10 and 22 postpartum. The thickness of the smooth muscle layers was found to decrease rapidly postpartum: the circular smooth muscle layer returned to that of a non‐pregnant, control uterus by day 5 postpartum and the longitudinal smooth muscle layer by day 15 postpartum. Eosinophilic cells were observed in the endometrial stroma adjacent to the myometrium on days 10, 15 and 20 postpartum, and were confirmed as collagenous cells. Immunostaining identified collagen type III positive cells in the vessel‐rich layer adjacent to the placental site on days 1, 3, 5 and 10 postpartum, and these cells were confirmed to be phagocytic. Postpartum reduction in the weight of the uterus was accompanied by decreases in both the collagen content and the thickness of the longitudinal and circular smooth muscle layers. Furthermore, the phagocytic cells were shown to express MPO during postpartum involution of the uterus.     

Th1/Th2 balance in canine peripheral blood lymphocytes--a flow cytometric study

The canine immune system undergoes continuous remodeling with advancing age. We measured the Th1/Th2 balance in peripheral blood lymphocytes (PBLs) obtained from 23 Beagles ranging in age from 0.5 to 11.8 years by flow cytometric analysis using intracellular cytokine staining. The percentage of CD4 cells producing interferon-gamma (Th1) increased with age. The percentage of CD4 cells producing interleukin-4 (Th2) increased but much less so. In conclusion, we demonstrated that the Th1/Th2 balance in canine peripheral blood could be measured by this flow cytometry technique and the Th1/Th2 balance inclined to dominance of the Th1 subpopulation in PBLs as the dog matured.