Authentication of flying-fish-meal content of processed food using PCR-RFLP

Nucleotide sequence and polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) analysis of the 3′-portion of the mitochondrial 16S RNA gene (rDNA) coding sequence were used to differentiate between flying fishes and other fishes that are frequently used in ago-noyaki production. In this study, we successfully distinguished between flying fishes and the other fishes by combining amplified DNA fragments with universally designed primers and digesting the PCR products with AfaI and MfeI restriction endonucleases. PCR-RFLP of 15 ago-noyaki, 2 noyaki, and 8 other processed flying-fish products were analyzed to authenticate flying-fish content among processed seafood products. After digestion of the PCR products with both enzymes, we found that all ago-noyaki and processed flying fish products contained either two or three DNA fragments of ~200, 300, and 530 bp, and noyaki samples (which do not contain flying fish) contained only one fragment of ~530 bp. Here, we present a new procedure to confirm the content of flying-fish meal in ago-noyaki.     

Approachability and contact behavior of commercial dairy calves to humans

Calves (n = 106) on four dairy farms were observed for their approachability to humans. All calves experienced similar rearing conditions: Beginning individual pen, after birth until weaning at about 2 months, where they were housed individually and fed milk and a milk replacement; Late individual pen, after weaning until grouping at about 3.5 months, where they were housed individually and fed hay, silage and concentrate feed; Beginning group pen, after grouping until 5 months, where they were housed in groups of 2–5 animals and fed hay, silage and concentrate feed; later group pen, from 5 to 7 months. The number of calves that contacted an experimenter who stood in front of their pens for 10 min was recorded on 6 separate days over 3 months. Latency to touch and time spent in activities during touching such as sucking, licking, biting and rubbing were also measured. There were no significant differences in the latency to touch and the ratio of touch to non‐touch calves between the rearing conditions and the farms. The time spent touching was significantly affected by the interaction between the rearing condition and the farm (P < 0.01). In detail, the time spent sucking (P < 0.001) and licking (P < 0.01) was different between the rearing condition × farm variables. The proportion of calves that approached and touched the experimenter tended to be higher in the farms in which a stockperson worked longer inside and outside their pens (both ρ = 0.95, P = 0.051). These results were interpreted according to the perspectives of early positive reinforcement with food and the habituation process to humans existing nearby.     

Isolation and characterization of the interspecific fusants from Streptomycetes obtained using a liquid regeneration method

ABSTRACT: Protoplast fusion between different species of Streptomyces was performed using a liquid regeneration method developed for a rapid and simple preparation of the fusants. Consequently, new clones, which could not be obtained using the conventional agar regeneration method, were obtained. In the crosses between S. griseus and S. durhamensis , and between S. californicus and S. catenulae , eight and two recombinants, respectively, were obtained using the liquid regeneration method. Conversely, in the case of crosses between S. ornatus and S. catenulae , and between S. ornatus and S. vendargensis , seven recombinants each were obtained using only the agar method. The physiological characteristics, such as the assimilation of carbohydrate and antibiotic resistance, of these fusants differed considerably from those of their parental strains. Using the proposed liquid regeneration method, a simpler and quicker procedure for protoplast fusion is described.     

Glomus tumor of the liver in a cow

An 11-year-old Holstein-Friesian cow exhibited anorexia and jaundice. A large mass was found in the liver during necropsy. Macroscopically, the mass was composed of dark red multilobular tissue and a centrally located abscess, which was connected to the hepatic duct. Histologically, the mass consisted of proliferation of small neoplastic cells and was demarcated from the hepatic parenchyma by a thick region of granulation tissue. The neoplastic cells were predominantly arranged in solid sheets, but they also formed blood-filled cancellous structures, and proliferating foci were seen around blood vessels. Periodic acid-Schiff reaction demonstrated that a fine basement membrane-like structure surrounded the neoplastic cells. Immunohistochemically, the neoplastic cells were positive for vimentin and alpha smooth muscle actin and negative for cytokeratin, factor VIII-related antigen, chromogranin and desmin. Based on its histopathological features, the hepatic neoplasm was diagnosed as a primary glomus tumor. This is the first report about a primary glomus tumor of the liver in a cow.     

Evaluation of S100B in cerebrospinal fluid as a potential biomarker for neurological diseases in calves

S100B in cerebrospinal fluid (CSF-S100B) was measured in calves with 20 neurologic and 21 non-neurologic diseases to clarify its utility as a biomarker for neurologic diseases. The median CSF-S100B value in the neurologic disease group (43.0 ng/ml) was significantly higher than that in the non-neurologic disease group (10.2 ng/ml). As CSF-S100B levels in calves with neurologic diseases widely differed, the utility of CSF-S100B as a diagnostic marker for neurologic diseases in cattle remains inconclusive.     

Detection of the nonsense mutation of OPA3 gene in Holstein Friesian cattle with dilated cardiomyopathy in Japan

Bovine dilated cardiomyopathy (DCM) is an autosomal recessive genetic disorder causing congestive heart failure and subsequent death. Recently, a nonsense mutation c.343C>T in the bovine optic atrophy 3 (OPA3) gene had been reported to cause the DCM in Holstein cattle in Switzerland. However, the mutation has not been confirmed in bovine DCM outside Switzerland. Nine Holstein Friesian cows that were macroscopically and histologically diagnosed with or suspected of DCM and 12 control cows kept in Japan were tested for the mutation. The mutation surrounding OPA3 DNA fragment was amplified by PCR and subjected to direct sequences. The homogeneous c.343C>T mutation was proved to occur in all the affected cows and not in the control cows. The present study is the first report of the mutation in the DCM affected cows outside Switzerland.     

Arrhythmogenic right ventricular cardiomyopathy coincided with the cardiac fibrosis in the inner muscle layer of the left ventricular wall in a boxer dog

A 7-year-old female boxer dog died suddenly without any clinical signs. It was suspected that the dog had arrhythmogenic right ventricular cardiomyopathy (ARVC) due to ventricular premature complexes and ventricular tachycardia at 3 years of age. The final diagnosis of ARVC was confirmed by histological characteristics, such as loss of cardiocytes and fibrofatty replacement, occurring in the right and left ventricular walls. In the cardiocytes, non-lipid vacuoles were observed. Cardiac fibrosis and intimal thickening of the small arteries occurred without fatty replacement in the inner muscle layer including the papillary muscles of the left ventricular wall. This paper describes the pathomorphological details of an ARVC case with coincidental cardiac fibrosis in the inner muscle layer of the left ventricular wall.     

Analysis of the bone morphogenetic protein 6 gene promoter region in young beef cattle affected by enzootic bovine leukosis

Enzootic bovine leukosis (EBL) is typically observed in cattle over 3 years old. However, some cases of EBL onset in young beef cattle have been reported in Japan. The mechanism for early EBL onset is unclear. In Japan, beef cattle are given large amounts of concentrated feed with low vitamin A. Bone morphogenetic proteins (BMPs) are regulators of cell proliferation, differentiation, and apoptosis, and thought to represent one of the key players in tumor malignancy. The purpose of this study was to evaluate the differences in BMP-6 methylation status between EBL beef cattle under 3 years old and other cattle. We investigated the methylation status of the BMP-6 promoter region in 32 EBL beef cattle under 3 years old. We also compared the methylation status of EBL dairy cattle to that of healthy cattle. Median methylation rate of the BMP-6 promoter region in EBL beef cattle under 3 years old was 8.9%, which was significantly higher than that of other groups. Hypermethylation of the BMP-6 promoter region might contribute to early onset of EBL in beef cattle under 3 years old, and animal feeding management practices specific to beef cattle may affect the methylation status of the BMP-6 promoter region.     

Genetic diversity of Sicyos angulatus in central and north-eastern Japan by inter-simple sequence repeat analysis

For the development of effective procedures to control invasive plants, it is necessary to learn about their mechanism of spread, for which an understanding of the plant's genetic variation may be important. Sicyos angulatus is a widespread and invasive weed that grows among forage crops and natural vegetation in Japan. Inter-simple sequence repeat (ISSR) genotyping was used to detect the regional patterns of genetic variation of S. angulatus in its introduced range in Japan. The goal of this research was to assess the introduction dynamics and mechanism of spread of S. angulatus in Japan. Four screened ISSR primers produced 15 reliable bands, and 12 of these were polymorphic among six areas in central and north-eastern Japan. The analysis of molecular variance ( amova ) revealed that 88.4% of genetic variation occurred within areas rather than between the two regions (1.1%), or among areas within regions (10.5%). While the total gene diversity among areas ( Ht ) was 0.1684, the coefficient of gene differentiation was low among two geographically distinct regions ( Gst = 0.053). The genetic diversity indices ( h  = 0.116; I  = 0.173) of the Fuji river area, assumed to be near the first introduction site, were not higher than other areas (total h  = 0.165; total I  = 0.263). In contrast, those of the Chikuma river and Tenryu river areas were relatively high ( h  = 0.173 and 0.187; I  = 0.276 and 0.277 respectively). The Mantel test showed no significant correlation between geographical and genetic distances. These results suggest multiple introductions from the same gene pools into both central and north-eastern regions of Japan.