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.     

Serum progesterone concentrations using P-EIA kit in captive and free-ranging Hokkaido brown bears, Ursus arctos yesoensis.

Serum progesterone (P) concentrations using P-EIA kit (Ovucheck, Cambridge Life Science Co., Ltd.) were examined in 8 captive and 7 free-ranging female Hokkaido brown bears (Ursus arctos yesoensis). The intra- and inter-assay coefficients of variation were 8.9%, 12.6% and 16.6%, 22.7%, respectively, based on 2 serum samples. There was a significant correlation between EIA and radioimmunoassay results based on 64 serum samples (r = 0.725; p less than 0.01). Serum P concentrations were examined in 5 pregnant, 2 solitary non-pregnant bears and a lactating non-pregnant bear in captivity. Annual changes of P levels in pregnant bears were observed as a small elevation during the mating season (May-June), a re-elevation in September-October and a sharp elevation in November-December. The sharp elevation was suspected to reflect changes when implantation occurred. Annual changes of P levels in solitary non-pregnant bears were similar to those in pregnant bears. An annual change of P levels in a lactating non-pregnant bear maintained levels under 5 ng/ml. Two of 7 free-ranging bears exhibited P levels over 1 ng/ml and the birth of cubs was confirmed in the following year in 1 of the 2 bears. P concentrations of other free-ranging bears exhibited less than 1 ng/ml, and these bears were considered to be non-pregnant. It was concluded that P-EIA kit was available for measuring P concentrations in Hokkaido brown bears.     

Genetic relationship of litter traits between farrowing and weaning in Landrace and Large White pigs

We estimated genetic parameters in Landrace and Large White pig populations for litter traits at farrowing (total number born, number born alive, number stillborn, total litter weight at birth (LWB), and mean litter weight at birth) and those at weaning (litter size at weaning (LSW), total litter weight at weaning (LWW), mean litter weight at weaning (MWW), and survival rate from farrowing to weaning). We analyzed 65,579 records at farrowing and 6,306 at weaning for Landrace, and 52,557 and 5,360, respectively, for Large White. Single‐trait and two‐trait repeatability animal models were exploited to estimate heritability and genetic correlation respectively. Heritability estimates of LSW were 0.09 for Landrace and 0.08 for Large White. Genetic correlations of LSW with MWW were –0.43 for Landrace and –0.24 for Large White. Genetic correlations of LSW with LWW and LWB ranged from 0.5 to 0.6. The genetic correlation of MWW with LWW was positive, but that with LWB was negligible. The results indicate that utilizing LWW or LWB could improve LSW efficiently, despite the antagonistic genetic correlation between LSW and MWW.     

Effect of negative dietary cation‐anion differences on carcass characteristics and beef tenderness of Japanese Black steers

Lowering dietary cation‐anion differences (DCAD) can enhance responsiveness to Ca‐homeostatic hormones and increase Ca availability, which might have potential to activate a Ca‐dependent protease, calpain, and to enhance postmortem myofibrillar proteolysis. In this study, we investigated the effects of DCAD manipulation on calpain activity and beef tenderness in Japanese Black cattle which are characterized by their high marbling. Thirty‐six Japanese Black steers were allotted to one of two treatments: (i) control (CON; DCAD +6.09 mEq/100 g of dry matter (DM)) or (ii) negative DCAD (NEGD; DCAD ?8.27 mEq/100 g DM) for 70 days before slaughter. Lowering DCAD decreased DM and energy intake (P < 0.01) even though it did not negatively affect the growth performance or carcass characteristics. In NEGD, urine pH was decreased by acidification caused by the negative DCAD (P < 0.01). Calpain activities tended to be improved in NEGD (P = 0.09), but Warner‐Bratzler shear force values were not affected by treatment. Although calpain activities tended to improve, lowering DCAD to ?8.27 for 70 days before slaughter was insufficient to enhance beef tenderness in Japanese Black steers.     

Genetic evaluation of the longevity of the Holstein population in Japan using a Weibull proportional hazard model

The objective of this study was to confirm the stability of the genetic estimation of longevity of the Holstein population in Japan. Data on the first 10 lactation periods were obtained from the Livestock Improvement Association of Japan. Longevity was defined as the number of days from first calving until culling or censoring. DATA1 and DATA2 included the survival records for the periods 1991–2003 and 1991–2005, respectively. The proportional hazard model included the effects of the region-parity-lactation stage-milk yield class, age at first calving, the herd–year–season, and sire. The heritabilities on an original scale of DATA1 and DATA2 were 0.119 and 0.123, respectively. The estimated transmitting abilities (ETAs) of young sires in DATA1 may have been underestimated, but coefficient δ, which indicated the bias of genetic trend between DATA1 and DATA2, was not significant. The regression coefficient of ETAs between DATA1 and DATA2 was very close to 1. The proportional hazard model could steadily estimate the ETA for longevity of the sires in Japan.     

Volatile organic compounds from wood and their influences on museum artifact materials II: Inference of causal substances of deterioration based on intercomparison of laser Raman spectra of deteriorated products

Volatile organic compounds (VOCs) from wood and their effects on artifacts are of intense interest to museum staff who must strictly manage the preservation environment. In this study, the causal substances of deterioration were inferred by comparison with laser Raman (LR) spectra of deterioration products. The artifact samples were deteriorated using VOCs from woods (WV test), and using specific substances that occur in wood VOCs (SV test). The deteriorated samples were analyzed by LR spectroscopy and the results of the WV and SV tests were compared. Hinokitiol and acetic acid were found to be two of the main causal substances of deterioration in western red cedar. Deterioration of iron and copper by spruce did not appear to be caused by acetic acid, as is generally assumed, but by some other unknown compound(s). Sensitivity to wood-based VOC components depended on the type of artifact. In western red cedar, mitsudasou (litharge) was very sensitive to acetic acid, while other artifacts were sensitive to hinokitiol. The LR method used in this study is very useful for the inference of causal substances of deterioration based on the detection of small amounts of deteriorated products such as those generated by deterioration due to wood VOCs. Part of this article was presented at the 25th Annual Meeting of the Japan Society for the Conservation of Cultural Property, Kyoto, June 2003     

Molecular cloning of tropomyosins identified as allergens in six species of crustaceans

Although tropomyosin is known to be a major allergen of crustaceans, its structural information is limited to only five species. In this study, tropomyosin was confirmed to be a major allergen in six species of crustaceans (black tiger prawn, kuruma prawn, pink shrimp, king crab, snow crab, and horsehair crab) by immunoblotting. Then, the amino acid sequences of tropomyosins from these crustaceans were elucidated by a cDNA cloning technique. Sequence data for crustacean tropomyosins including the obtained results reveal that fast tropomyosins are contained in shrimps (or prawns) and lobsters, slow tropomyosins in crabs, and both tropomyosins in crayfishes and hermit crabs. Although fast and slow tropomyosins share a high sequence identity (about 90%) with each other, significant differences are observed in specific regions between both tropomyosins.     

Comprehensive analytical method for the determination of hydrophilic metabolites by high-performance liquid chromatography and mass spectrometry

A method for the comprehensive analysis of hydrophilic metabolites, based on a combination of high-performance liquid chromatography and mass spectrometry, is described. We evaluated three types of stationary phases to achieve the separation of highly hydrophilic metabolites. Good chromatographic retention and separation of these metabolites were achieved on a pentafluorophenylpropyl-bonded silica column with gradient elution, using 0.1% aqueous formic acid and acetonitrile as the mobile phase. The optimized conditions allowed the comprehensive determination of the standard 49 kinds of amino acids, 6 kinds of amines, 45 kinds of organic acids, 18 kinds of nucleic bases, 5 kinds of nucleosides, and 14 kinds of nucleotides, and then the linearity, dynamic range, detection limit, and precision of the retention time and the peak area were validated. We applied this method for the targeted analysis of the components in soy sauce. The results from the quantitative determination of amino acids were compared to those obtained with an amino acid analyzer, and the accuracy was in the range between 85 and 119%. The accuracy of other detected components was confirmed to be 105-133% by the recovery rate after the addition of standard compounds. We also applied the method for the nontargeted metabolic profiling of the components in several kinds of soy sauces with the principal component analysis. They were classified by the manufacturing methods, and the components that corresponded to the differences were identified. This method could be useful for the targeted analysis of hydrophilic metabolites as well as their nontargeted metabolic profiling.     

Radical scavenging and singlet oxygen quenching activity of marine carotenoid fucoxanthin and its metabolites

Antioxidant activity of carotenoids is suggested to be one of the factors for their disease preventing effects. Marine carotenoids fucoxanthin and its two metabolites, fucoxanthinol and halocynthiaxanthin, have been shown to exhibit several biological effects. The antioxidant activities of these three carotenoids were assessed in vitro with respect to radical scavenging and singlet oxygen quenching abilities. The 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity of fucoxanthin and fucoxanthinol was higher than that of halocynthiaxanthin, with the effective concentration for 50% scavenging (EC 50) being 164.60, 153.78, and 826.39 microM, respectively. 2,2'-Azinobis-3-ethylbenzo thizoline-6-sulphonate radical scavenging activity of fucoxanthinol (EC 50, 2.49 microM) was stronger than that of fucoxanthin (EC 50, 8.94 microM). Hydroxyl radical scavenging activity as measured by the chemiluminescence technique showed that the scavenging activity of fucoxanthin was 7.9 times higher than that by fucoxanthinol, 16.3 times higher than that by halocynthiaxanthin, and 13.5 times higher than that by alpha-tocopherol. A similar trend was observed when the hydroxyl radical scavenging was assessed by the electron spin resonance (ESR) technique. ESR analysis of the superoxide radical scavenging activity also showed the superiority of fucoxanthin over the other two carotenoids tested. Singlet oxygen quenching ability of the three carotenoids was lower than that of beta-carotene, with quenching rate constants ( k Q, x10 (10) M (-1) s (-1)) being 1.19, 1.81, 0.80, and 12.78 for fucoxanthin, fucoxanthinol, halocynthiaxanthin, and beta-carotene, respectively. The higher radical scavenging activity of fucoxanthin and fucoxanthinol compared with halocynthiaxanthin is assumed to be due to presence of the allenic bond.