Decreased transport of orally administered protein into the blood circulation of developing juveniles of Japanese eel Anguilla japonica

ABSTRACT: To clarify the quantitative changes in the transport of orally intubated protein into the blood circulation as macromolecules in development, immunoglobulin Y (IgY) extracted from chicken eggs was administered orally to juvenile Japanese eel, Anguilla japonica . For the first experiment, which was performed before the commencement of artificial feeding, the oral delivery of 2.0 μg/0.1 g bodyweight of IgY resulted in a rapid increase in plasma IgY to a maximum of 2.30 μg/mL. However, the transport of IgY into the blood decreased significantly in the experiments that followed, which were performed after 12, 25 and 42 days. During this period, bodyweight increased approximately by a factor of eight, and rapid growth of the stomach was observed histologically. Possible contributions for the development of the alimentary canal to the diminishment of intestinal protein assimilation are discussed.     

Bacterial leaf blight of sweet pepper (Capsicum annuum) caused by Pseudomonas cichorii in Japan

Sweet peppers (cv. Tosajishi beauty) with leaf blight symptoms were observed in Kochi Prefecture, 2003. Initially, small spots formed on leaves, and later enlarged to brown to dark brown spots, and eventually blighted leaves fell. Several bacteria that caused the same symptoms were isolated and subsequently reisolated. These bacteria were identified as Pseudomonas cichorii on the basis of bacterial characteristics and the 16S rRNA gene sequence. This is the first report of bacterial leaf blight in the sweet pepper in the world.     

Effect of environmental factors on fertility of Holstein–Friesian cattle in Japan

We investigated the environmental factors in Japan, including meteorological conditions, on the fertility of a European cattle breed, Holstein–Friesian, by examining conception rates in different regions. First artificial insemination and associated conception details were recorded for 69,952 Holstein female cattle. In general, meteorological conditions vary considerably according to latitude in Japanese islands. Conception rates were higher in the Northern (above 37°N) than the Southern (below 37°N) region (61.3% vs. 53.3%). All the factors analyzed in the statistical model, including insemination year, region, month, AI technician, service sire and interaction between region and month, had significant effects on the conception rate. In the Southern region, conception rates were lower in the summer months (average temperature, 27.8 °C, and maximum temperature, 32.3 °C). However, this seasonal decline was not observed in the Northern region (average temperature, 23.7 °C, and maximum temperature, 28.4 °C). Regression analysis of conception rate in relation to temperature showed highly significant negative regression coefficients (− 0.66 to − 0.63% per °C; p < 0.001) in the Southern region, but non-significant coefficients in the Northern region. Humidity had a minimal effect on conception rate in both regions. Our data collectively suggest that 37°N latitude is a threshold that affects Holstein–Friesian conception rates in Japan.     

Establishment of a potency test by ELISA for a rabies vaccine for animal use in Japan

The ELISA we developed was able to determine the antigen content and was suitable for a potency test, and we described a relative potency assay method which determines the potency of test vaccines by comparing the ELISA value of a test vaccine to that of a reference vaccine. In the present study, we standardized the reference vaccine used for determining the potencies of test vaccines, and established a potency test by ELISA. We evaluated the proposed reference vaccine by the neutralizing antibody responses in dogs after vaccination, by the challenge protection test in guinea pigs (GP potency test), which is the earlier official potency test used in Japan, and by the NIH potency test, which is widely used throughout the world. The results showed that a 4-fold dilution of the proposed reference vaccine induced sufficient immunity in dogs. A 3-fold dilution of the proposed reference vaccine passed the GP potency test. The international units (IU) calibrated by the NIH potency test were 3.7 IU/dose. From the results and the WHO recommendation that veterinary rabies vaccines should have a potency of at least 1.0 IU/dose, we determined to dilute the proposed reference vaccine by 3 fold and regarded it as the reference vaccine. Finally, we confirmed that there is a good agreement between the results of the potency test by ELISA and the results of the GP potency test. The establishment of the potency test by ELISA has made it possible to monitor the potency in the production process and has contributed to the stable production of the vaccine.     

Antibody response to toxic shock syndrome toxin-1 of Staphylococcus aureus in dairy cows

Antibody response to toxic shock syndrome toxin-1 (TSST-1) of Staphylococcus aureus in dairy cows was examined by enzyme linked immunosorbent assay (ELISA). Serum antibody to TSST-1 was not detected in 39 (76.5%) of 51 calves, which were 1-6 months of age. In contrast, TSST-1 antibody was demonstrated in 1728 (72.6%) of 2380 lactating cows housed on 36 dairy farms. The ELISA values of antibody ranged from 0.2 to 3.0 OD and presented a distribution with the peak at 1.6 OD. The mean ELISA value differed between farms, and it increased slightly along with parturient history. Somatic cell counts of milk from 174 lactating cows was compared with TSST-1 antibody and tst1,000,000 cells per ml. The mean ELISA values in milk were lower than those of sera, but they rose as somatic cells increased. The tst gene of S. aureus detected in 76.0-86.2% of the milk samples containing somatic cells > 500,000 cells per ml, a level which indicates mastitis. The data suggests that many lactating cows may be infected by TSST-1- producing S. aureus.     

Characterization of a novel nicotinamide adenine dinucleotide-cytochrome b5 reductase mutation associated with canine hereditary methemoglobinemia

Hereditary methemoglobinemia associated with nicotinamide adenine dinucleotide-cytochrome b5 reductase (b5R) deficiency is a rare autosomal recessive disorder in animals. Recently, nonsynonymous b5R gene (CYB5R3) variants have been reported to be associated with canine and feline hereditary methemoglobinemia. However, the underlying molecular mechanisms of canine and feline methemoglobinemia caused by these nonsynonymous variants have not yet been reported. Previously, we reported a Pomeranian dog family with hereditary methemoglobinemia, carrying CYB5R3 mutation of an A>C transition at codon 194 in exon 7, replacing an isoleucine residue with leucine (p.Ile194Leu). In this study, we investigated the enzymatic and structural properties of the soluble form of wild-type and Ile194Leu canine b5Rs to characterize the effects of this missense mutation. Our results showed that the kinetic properties of the mutant enzyme were not affected by this amino acid substitution. The secondary structure of the wild-type and Ile194Leu b5Rs detected by circular dichroism showed a similar pattern. However, the mutant enzyme exhibited decreased heat stability and increased susceptibility to trypsin hydrolysis. Moreover, the thermostability and unfolding measurements indicated that the mutant enzyme was more sensitive to temperature-dependent denaturation than the wild-type b5R. We concluded from these results that unstable mutant enzyme properties with normal enzymatic activity would be associated with hereditary methemoglobinemia in the Pomeranian dog family.     

Classification of abomasal displacement in cows according to histopathology of the liver and clinical chemistry

Histopathological features of livers and blood chemical values in cows with abomasal displacement were investigated. Liver biopsy samples were collected during redressment operations in 92 cows with abomasal displacement, and the samples were stained with haematoxylin and eosin or periodic acid Schiff (PAS). Blood was collected for chemical tests. Livers were histopathologically divided into the following four types: normal histology cases (21%), fatty degeneration cases (36%), cloudy swelling cases (19%) and fatty degeneration cases with cloudy swelling (24%). The number of PAS-positive samples was significantly higher in the normal histology group and significantly lower in the severe fatty degeneration group and severe cloudy swelling group. Cows with fatty degeneration had significantly higher levels of serum 3-hydroxybutyric acid, non-esterified fatty acid and aspartate aminotransferase than did those with cloudy swelling or normal histology. The results indicate that the morbid conditions of cows with abomasal displacement can be classified into four types.     

Suppressive effect of deoxynivalenol, a Fusarium mycotoxin, on bovine and porcine neutrophil chemiluminescence: an in vitro study

We evaluated the immunotoxicity of deoxynivalenol (DON), a Fusarium mycotoxin, on bovine and porcine neutrophils in vitro by using two function parameters, luminol-dependent chemiluminescence and random migration under agarose. A 2-hr DON treatment suppressed the chemiluminescence of bovine and porcine cells by 42% and 35% (on average) at 10(-5)M, and by 19% and 26% at 10(-6)M. Slight suppression was observed at concentrations lower than 10(-6)M. However, after an 18-hr DON treatment, random migration of neutrophils of both species remained unaffected, even at the highest concentration (10(-5)M). Although further extensive studies are needed, to our knowledge this is the first study to have revealed in vitro that DON can affect neutrophil function.     

Retrospective diagnosis of feline GM2 gangliosidosis variant 0 (Sandhoff-like disease) in Japan: possible spread of the mutant allele in the Japanese domestic cat population

GM2 gangliosidosis variant 0 (human Sandhoff disease) is a lysosomal storage disease caused by simultaneous deficiencies of acid beta-hexosaminidase (Hex) A and Hex B due to an abnormality of beta-subunit, a common component in these enzyme molecules, which is coded by the HEXB gene. In the present study, a retrospective diagnosis was performed in 2 previous suspected cases of feline Sandhoff-like disease using a DNA test to detect the causative mutation identified previously in 4 cats in 2 other families of Japanese domestic cats. Enzymic analysis was also performed using stored leukocytes and plasma collected from the subject families in order to investigate the usefulness of enzymic diagnosis and genotyping of carriers. The DNA test suggested that the 2 cases were homozygous recessive for the mutation. Consequently, 6 cats homozygous for the same mutation have been found in 4 separate locations of Japan, suggesting that this mutant allele may be spread widely in the Japanese domestic cat populations. In enzymic analysis, Hex A and Hex B activities in leukocytes and plasma measured using 4-methylumbelliferyl N-acetyl-beta-D-glucosaminide as a substrate were negligible in affected cats, compared with those in normal and carrier cats. However, there was a wide overlap in enzyme activity between normal and carrier cats. Therefore, it was concluded that enzymic analysis is useful for diagnosis of affected cats, but is not acceptable for genotyping of carriers.