A survey on the prevalence and distribution of antibodies to BLV was performed by the agar-gel immunodiffusion test over a period from 1983 to 1985. More than 2,407 serum samples were collected from Holstein cattle raised in the eastern part of Saitama prefecture where suburban dairy farm is operated. The average positive rate of this period was 4.9 %. The rates of reactive samples varied from 2.6 to 9.8 % among the age groups of cattle from younger than one year to 14 years of age. The positive rate increased gradually with age. The positive rates also varied widely from 0 to 21 % among areas surveyed. Furthermore, there were large differences in this rate among farms even in the same area. The results were interpreted and discussed in connection with the enzootic feature of BLV infection.

Inflammatory mammary carcinoma (IMC) is a rare disease with a poor prognosis and one affecting dogs. Inflammatory breast carcinoma (IBC) is a subtype of malignant breast cancer in humans with a high degree of malignancy and a similarly poor prognosis. Since the clinical symptoms and prognoses of both are similar, canine IMC has been considered as a model of human IBC. In this study, we newly established a stable IMC-derived cell line from a patient at the Yamaguchi University Animal Medical Center in Japan. The patient was a female toy poodle presenting with an inflamed mammary gland, which was diagnosed as IMC. The cell line was established from a tissue biopsy. Surface antigen marker (CD24 and CD44) expression was determined. Cystine/glutamate antiporter (xCT) expression was determined by Western blotting, flow cytometry and fluorescence immunostaining, and sulfasalazine was administered to ascertain if it suppressed xCT expression. Stem cell marker (Nanog, Sox2, Myc and Klf4) expression and aldehyde dehydrogenase (ALDH) activity were also investigated. The cultured cells showed xCT, and its suppression showed downregulation of stem cell markers and ALDH activity. Stable cell proliferation was verified. A new canine IMC-derived cell line was established. In the future, we aim to study the effect of xCT on the maintenance of cancer stem cell properties in canine tumours, and propose a new therapeutic method for the treatment of canine IMC by targeting xCT.

Seal parapoxvirus (SPPV) infection has been reported among pinnipeds in aquaria in Japan; however, its seroprevalence is unknown. Therefore, an enzyme-linked immunosorbent assay (ELISA) was developed for serological diagnosis of SPPV infection. The gene encoding the major envelope protein of SPPV was cloned into the eukaryotic expression vector pAcGFP1-N1, which encodes the green fluorescence protein (GFP), thereby producing a fusion protein (Env-GFP). Parental and cloned vector DNA was independently transfected into cultured seal cells for the expression of GFP and Env-GFP. The wells of an ELISA plate were coated with either GFP- or Env-GFP-transfected cell lysates. The light absorbance of each serum sample was adjusted by subtracting the absorbance of GFP-coated wells from that of Env-GFP-coated wells. Sera from two spotted seals (Phoca largha), six beluga whales (Delphinapterus leucas), three Pacific white-sided dolphins (Lagenorhynchus obliquidens), and ten bottlenose dolphins (Tursiops truncatus) from an aquarium in Japan were examined using the ELISA. Positive reactions were not observed, except in one preserved sample collected ten years ago from a naturally SPPV-infected spotted seal. The established ELISA could be useful in screening marine mammal sera for anti-SPPV antibodies.

Land application of manure that contains antibiotics and resistant bacteria may facilitate the establishment of an environmental reservoir of antibiotic-resistant microbes, promoting their dissemination into agricultural and natural habitats. The main objective of this study was to search for acquired antibiotic resistance determinants in the gut microbiota of wild boar populations living in natural habitats. Gastrointestinal samples of free-living wild boars were collected in the Zemplén Mountains in Hungary and were characterised by culture-based, metagenomic, and molecular microbiological methods. Bioinformatic analysis of the faecal microbiome of a hunted wild boar from Japan was used for comparative studies. Also, shotgun metagenomic sequencing data of two untreated sewage wastewater samples from North Pest (Hungary) from 2016 were analysed by bioinformatic methods. Minimum spanning tree diagrams for seven-gene MLST profiles of 104 E. coli strains isolated in Europe from wild boars and domestic pigs were generated in Enterobase. In the ileum of a diarrhoeic boar, a dominant E. coli O112ab:H2 strain with intermediate resistance to gentamicin, tobramycin, and amikacin was identified, displaying sequence type ST388 and harbouring the EAST1 toxin astA gene. Metagenomic analyses of the colon and rectum digesta revealed the presence of the tetQ, tetW, tetO, and mefA antibiotic resistance genes that were also detected in the gut microbiome of four other wild boars from the mountains. Furthermore, the tetQ and cfxA genes were identified in the faecal microbiome of a hunted wild boar from Japan. The gastrointestinal microbiota of the free-living wild boars examined in this study carried acquired antibiotic resistance determinants that are highly prevalent among domestic livestock populations.

Introduction: Breeding profiles at the periparturient stage in red foxes which mated naturally or were subjected to artificial insemination were retrospectively surveyed using 130 vixens during their reproductive seasons of 2012–2017 in Japan. Material and Methods: Natural mating vixens were encouraged a maximum of three times with the same male, while artificial insemination was conducted using frozen-thawed semen with the bovine semen extender as a diluent. Results: With natural mating, conception rates after one, two, and three copulations were 55.8%, 68.0%, and 85.7%, respectively, showing a significant difference between the rates for one and three copulations. Conception rates with artificial insemination were 82.4%. Mean gestation periods were between 52.1 and 53.3 days in all groups. Mean litter sizes were 3.7–4.3 cubs with natural mating, and 4.4 cubs with artificial insemination. Although some sporadic and inconsistent changes in litter sizes were noted between primiparous and multiparous groups, these were of doubtful clinical importance. Conclusion: This is the first report from Japan concerning basic breeding events of red fox vixens in captivity.

Tetrodotoxin (TTX) is a toxin mainly occurring naturally in contaminated puffer fish, which are a culinary delicacy in Japan. It is also detected in various marine organisms like globefish, starfish, sunfish, stars, frogs, crabs, snails, Australian blue-ringed octopuses, and bivalve molluscs. TTX is produced by marine bacteria that are consumed mainly by fish of the Tetraodontidae family and other aquatic animals. TTX poisoning through consuming marine snails has recently begun to occur over a wider geographical extent through Taiwan, China, and Europe. This neurotoxin causes food intoxication and poses an acute risk to public health. The aim of this review is to present the most recent information about TTX and its analogues with particular regard to toxicity, methods of analysis, and risk to humans of exposure.

The aim of this study was to characterize peripartum metabolic profiles, including the insulin sensitivity index, in cows diagnosed with subclinical ketosis (SCK) in the early stage of lactation. Cows that calved from January 2011 through December 2014 on a dairy farm with alarm level prevalence of SCK in Hokkaido, Japan (n = 175) were used. Blood and body condition scores (BCS) were obtained at regular health examinations in 2 consecutive periods, the first between 14 and 2 d before parturition, and the second between 3 and 14 d after parturition. Animals were divided into 3 groups at postpartum sampling: an SCK group with 35 multiparous and 15 primiparous cows having β-hydroxybutyrate (BHBA) concentrations ≥ 1.2 mM without clinical signs, a disease group of 36 multiparous and 9 primiparous cows that received treatment between parturition and postpartum sampling, and a control group consisting of 49 multiparous and 31 primiparous cows with BHBA concentrations < 1.2 mM. The prepartum revised quantitative insulin sensitivity check index was significantly lower in the multiparous SCK and disease groups than in the control group, demonstrating decreased insulin sensitivity in these cows, but not in primiparous cows. The prepartum BCS was significantly higher only in the multiparous SCK and disease groups. The prepartum apolipoprotein B-100 (ApoB-100) concentration was significantly decreased in the multiparous disease group, suggesting hepatic lipidosis. Conversely, primiparous cows had a higher prepartum ApoB-100 concentration. Prepartum decreased insulin sensitivity in the multiparous SCK and disease groups was considered to facilitate progression to SCK after calving.

OBJECTIVE To evaluate colonization of Streptococcus suis and Streptococcus parasuis on pig farms in Japan and to identify sources of infections. SAMPLE Saliva, feces, and vaginal swab samples from 84 healthy pigs of several growth stages on 4 farms and swab samples of feed troughs and water dispensers at those farms. PROCEDURES Samples were collected from August 2015 to June 2016. Two quantitative PCR (qPCR) assays (one for S suis and the other for S parasuis) were designed for use in the study. The novel qPCR assays were used in combination with previously described qPCR assays for S suis serotype 2 or 1/2 and total bacteria. Relative abundance of bacteria in each sample was evaluated. RESULTS Streptococcus suis was detected in all saliva samples and some of the other samples, whereas S parasuis was not detected in any of the samples, including saliva samples, which indicated a difference in colonization preference. The ratio of S suis to total bacteria in saliva appeared to increase with age of pigs. Streptococcus suis serotype 2 or 1/2 was detected in a few saliva samples and feed trough swab samples at 2 farms where S suis infections were prevalent. CONCLUSIONS AND CLINICAL RELEVANCE Saliva, especially that of sows, appeared to be a reservoir and source of S suis infection for pigs. The qPCR assay described here may provide an effective way to monitor for S suis in live pigs, which could lead to effective disease control on pig farms.

Objective: To assess the status of antimicrobial resistance (AMR), identify extraintestinal virulence factors (VFs) and phylogenetic origins, and analyze relationships among these traits in extraintestinal pathogenic Escherichia coli (ExPEC) isolates from companion animals. Sample: 104 E coli isolates obtained from urine or genital swab samples collected between 2003 and 2010 from 85 dogs and 19 cats with urogenital infections in Japan. Procedures: Antimicrobial susceptibility of isolates was determined by use of the agar dilution method; a multiplex PCR assay was used for VF gene detection and phylogenetic group assessment. Genetic diversity was evaluated via randomly amplified polymorphic DNA analysis. Results: Of the 104 isolates, 45 (43.3%) were resistant to > 2 antimicrobials. Phylogenetically, 64 (61.5%), 22 (21.2%), 13 (12.5%), and 5 (4.8%) isolates belonged to groups B2, D, B1, and A, respectively. Compared with other groups, group B2 isolates were less resistant to all tested antimicrobials and carried the pap, hly, and cnf genes with higher frequency and the aer gene with lower frequency. The aer gene was directly associated and the pap, sfa, hly, and cnf genes were inversely associated with AMR. Randomly amplified polymorphic DNA analysis revealed 3 major clusters, comprised mainly of group B1, B2, and D isolates; 2 subclusters of group B2 isolates had different VF and AMR status. Conclusions and Clinical Relevance Prevalences of multidrug resistance and human-like phylogenetic origins among ExPEC isolates from companion animals in Japan were high. It is suggested that VFs, phylogenetic origins, and genetic diversity are significantly associated with AMR in ExPEC.