Nationwide survey of hepatitis E virus infection among wildlife in Japan

In Japan, hepatitis E virus (HEV) causes hepatitis in humans through the consumption of raw or undercooked meat, including game meat. In the present study, nationwide surveillance of HEV infection among a total of 5,557 wild animals, including 15 species, was conducted in Japan. The prevalence of anti-HEV antibodies in wild boar was 12.4%, with higher positive rates in big boars (over 50 kg, 18.4%) than in small individuals (less than 30 kg, 5.3%). Furthermore, HEV RNA was more frequently detected in piglets than in older boars. Interestingly, the detection of HEV among wildlife by ELISA and RT-PCR suggested that HEV infection in Sika deer was a very rare event, and that there was no HEV infection among wild animals except for wild boar, Sika deer and Japanese monkeys. In conclusion, wild boar, especially piglets, are at high risk of HEV infection, while other wild animals showed less risk or no risk of HEV transmission.     

Experimental hyperlipemia induces insulin resistance in cats

The effect of experimental hyperlipemia on insulin sensitivity was evaluated in seven healthy cats. Serum triglyceride and free fatty acid concentrations were significantly (P<0.05) higher when lipid-heparin was administered (2,894 ± 1,526 mg/dl and 4.54 ± 0.70 mEq/l, respectively) than when saline was administered (70 ± 42 mg/dl and 0.22 ± 0.08 mEq/l, respectively). A glucose clamp test revealed that the mean glucose infusion rate when lipid-heparin was administered (5.80 ± 0.67 mg/kg/min) was significantly (P<0.05) lower than when saline was administered (8.52 ± 1.83 mg/kg/min). These results suggest that experimental hyperlipemia induced insulin resistance in the healthy cats.     

Advanced maternal age induces fetal growth restriction through decreased placental inflammatory cytokine expression and immune cell accumulation in mice

Advanced maternal age is a risk factor for female infertility, and placental dysfunction is considered one of the causes of pregnancy complications. We investigated the effects of advanced maternal aging on pregnancy outcomes and placental senescence. Female pregnant mice were separated into three groups: young (3 months old), middle (8–9 months old), and aged (11–13 months old). Although the body weights of young and middle dams gradually increased during pregnancy, the body weight of aged dams only increased slightly. The placental weight and resorption rate were significantly higher, and live fetal weights were reduced in a maternal age-dependent manner. Although mRNA expression of senescence regulatory factors (p16 and p21) increased in the spleen of aged dams, mRNA expression of p16 did not change and that of p21 was reduced in the placenta of aged dams. Using a cytokine array of proteins extracted from placental tissues, the expression of various types of senescence-associated secretory phenotype (SASP) factors was decreased in aged dams compared with young and middle dams. The aged maternal placenta showed reduced immune cell accumulation compared with the young placenta. Our present results suggest that models using pregnant mice older than 8 months are more suitable for verifying older human pregnancies. These findings suggest that general cellular senescence programs may not be included in the placenta and that placental functions, including SASP production and immune cell accumulation, gradually decrease in a maternal age-dependent manner, resulting in a higher rate of pregnancy complications.     

Molecular and antigenic similarities of the fimbrial major components between Porphyromonas gulae and P. gingivalis

Porphyromonas gulae is black-pigmented anaerobic bacteria associated with canine periodontitis. There is little information available about the specific identify and relative occurrence of pigmented anaerobes in companion animals. Our aim was to clarify the factor involved in the adherence and colonization of the organism in the oral cavity. Fimbrial protein was purified from P. gulae ATCC 51700. The molecular mass of this protein was approximately 41kDa as estimated by SDS-PAGE. An antibody against 41-kDa fimbrial protein from P. gingivalis ATCC 33277 reacted with fimbrillin of P. gulae ATCC 51700. Immunogold electron microscopy revealed that the anti-41kDa fimbrial serum bound to fimbria on the cell surface of P. gulae ATCC 51700. Thus, fimbrial protein of P. gulae ATCC 51700 had the same size and antigenicity as 41-kDa fimbriae of P. gingivalis ATCC 33277. The nucleotide sequence of the fimA gene from P. gulae ATCC 51700 showed 94% homology with that of P. gingivalis ATCC 33277. Moreover, the deduced amino acid sequences have 96.8% identity. P. gulae has adherent ability to gingival epithelial cells. The properties of P. gulae fimbriae are similar to those of P. gingivalis fimbriae. We suggest that the surface structure of P. gulae may play a role in the colonization of this organism in periodontal pockets in companion animals.     

Establishment of a new canine cell line (CCT) originated from a cutaneous malignant histiocytosis

A new canine cell line, named CCT, was established from the cutaneous malignant histiocytosis in a 4-year-old male Borzoi. CCT proliferated with loose adherence and doubling time was approximately 30 hr. When co-cultured with latex beads, CCT phagocytized beads vigorously. Lysozyme and vimentin were positive by immunostaining, and non-specific esterase and acid phosphatase were positive by cytochemical staining. These features indicated the cells had a histiocytic nature. Furthermore, by subcutaneous injection to nude mice CCT could successfully form tumors with the morphological and immunohistochemical features similar to the original tumor.     

A comparative pathological study on canine necrotizing meningoencephalitis and granulomatous meningoencephalomyelitis

Canine necrotizing meningoencephalitis (NME) and granulomatous meningoencephalomyelitis (GME) were compared pathologically. Gross observation exhibited lateral ventricular dilation and discoloration, malacia and/or cavitation of the cerebrum in NME. On the contrary, gross changes were milder in GME, except for occasional visible granulomatous mass formation. Histopathologically, the lesions of NME were distributed predominantly in the cerebral cortex and various degrees of inflammatory and necrotic changes were observed according to clinical stages. Besides, microscopic lesions of GME were mainly distributed in the white matter of the cerebrum, cerebellum and brainstem, which are characterized by perivascular cuffing, multiple granulomas and leptomeningeal infiltrates. Although macrophages and lymphocytes were predominant in the inflammatory lesions of both disorders, macrophages in GME transformed into epithelioid cells and exhibited more massive infiltration. Although lectin RCA-1-reactive cells were numerous in both disorders, lysozyme immunoreactive cells in NME were fewer than that in GME. Parenchymal infiltration of MAC387-positive cells was common in GME and limited in NME. The number of CD3-positive lymphocytes in the GME lesions tended to be greater than in NME, though the difference was not statistically significant. Morphological and immunohistochemical differences of the lesions, in particular, the characteristics of infiltrative macrophages may reflect these different pathogeneses of the two disorders.     

Establishment of a Conditional Transgenic System Using the 2A Peptide in the Female Mouse Germline

Transgenic mice are essential research tools in developmental biology studies. The 2A peptide allows multiple genes to be expressed simultaneously at comparable levels in somatic cells, but there are no reports of it being used successfully in germ cells. We constructed a Cre/loxP-based conditional vector containing the 2A peptide to significantly enhance the expression of a reporter and target gene from a constitutive promoter in oocytes. Mice with a transgene insertion containing the chicken β-actin promoter, floxed EGFP-polyA cassette, mCherry reporter, 2A peptide and target gene DNA methyltransferase 3A2 (Dnmt3a2) were crossed with TNAP- or Vasa-Cre mice to produce offspring, in which mCherry and DNMT3A2 proteins were highly expressed in oocytes upon Cre-mediated removal of EGFP-polyA. This novel transgenic mouse line based on the 2A expression system can serve as a useful tool for examining gene function during oogenesis.     

Detection of CAP59 gene in 2 feline cases of systemic cryptococcosis

Two feline cases were diagnosed as systemic cryptococcosis due to Cryptococcus neoformans (teleomorph: Filobasidiella neoformans) by PCR assay with CAP59 gene primers using urine, serum and biopsy samples. The results of molecular analysis were consistent with the mycological findings.     

Differences between carcass grades in some morphological, behavioral and physiological measurements at early and middle stages of fattening in crossbred steers

The differential variables for carcass grades were identified from morphological, behavioral and physiological measurements of young steers at a commercial farm. Thirty‐five Japanese Black × Holstein steers aged 6–10 months were randomly assigned to three pens. The steers had free access to ryegrass hay and a restricted amount of concentrate diet for the first 6 months of trial. They were then given oat straw ad libitum and an additional concentrate diet until slaughter. Behavioral observations (15 categories) were carried out once every 2 weeks for 2 h after feeding. Bodyweight was measured, blood sampled (for seven metabolic hormones and five metabolites), ultrasonic scanning conducted and physical measurements taken (10 parts for each animal) in months 1, 3 and 5 (early fattening stage, EFS) and in months 7, 9 and 11 (middle fattening stage, MFS) of the trial. Temperament scores during these procedures, entry order into restraint stalls, social rank and average daily gain were also assessed. A factor analysis and one‐way anova were used to identify clusters of variables which had different factor scores between carcass grades. As for the carcass yields, a cluster of chest girth and depth, bodyweight, withers and hip height, and rump length (P < 0.01), and a cluster of the frequency of scratching the body with facilities (P < 0.05) were identified as differential variables in the EFS. In the MFS, a cluster of concentrations of cortisol and epinephrine, and the frequency of eating straw were identified (P < 0.10). As for the carcass quality grade, a cluster of social rank, triglyceride concentration and the frequencies of stand‐chewing cud and eating hay, and a cluster of the frequency of investigating facilities (both P < 0.10) were identified in the EFS. In the MFS, a cluster of hip height, bodyweight, cannon circumference, chest depth, withers height and chest girth, and a cluster of the frequencies of eating a concentrate diet and stand‐chewing the cud, and social rank (both P < 0.10) were identified. In conclusion, bodyweight, chest girth and depth, withers and hip height were identified as good differential variables for future carcass grades of young steers. Facility enrichment that encourages steers to eat hay in the EFS, and to eat a concentrate diet in and after the MFS would be effective in upgrading carcass grades. High incidence of investigating and scratching the body with facilities and stand‐chewing the cud might lead to lower carcass grades.