Indices using external measurements for assessing fat deposition of adult feral raccoons (Procyon lotor) in Kanagawa Prefecture, Japan

We examined the use of external measurements and relative fat deposition of adult feral raccoons (Procyon lotor) to develop relative indices of body fat deposition in post-growth feral raccoons. From March 2006 to March 2010, 288 adult raccoon carcasses (110 males, 178 females) collected in Kanagawa Prefecture, Japan, which were determined to be 24 months old, were subjected to external measurements of body weight (BW), girth measurement (GM), and body mass index (BMI). To assess relative body fat deposition, we visually classified abdominal subcutaneous fat into three grades (Visible Fat Index [VFI]: I-III). Significant differences in the means of BW (both sexes:P<0.01), GM (females: P<0.05, males: P<0.01), and BMI (both sexes: P<0.01) were detected between seasons. Notably, the means of BW, GM, and BMI (all, both sexes: P<0.01) differed significantly between VFI grades. However, by discriminant analysis with BW, GM, and BMI as independent variables, we obtained a significant discriminant function (both sexes: P<0.01) for distinguishing VFI I from higher VFI grades, but no significant equation was obtained for distinguishing between VFI II and VFI III. Based on the obtained structure matrix of discriminant analysis, BMI was the most valuable component for the discrimination of VFI grades. Thus, we conclude that BMI is a suitable complementary index for assessing relative body fat deposition of adult feral raccoons in Kanagawa Prefecture and may be generalizable to populations in other areas.     

Development of a pen-site test kit for the rapid diagnosis of H7 highly pathogenic avian influenza

As well as H5 highly pathogenic avian influenza viruses (HPAIV), H7 HPAIV strains have caused serious damages in poultry industries worldwide. Cases of bird-to-human transmission of H7 HPAIV have also been reported [11]. On the outbreak of avian influenza, rapid diagnosis is critical not only for the control of HPAI but also for human health. In the present study, a rapid diagnosis kit based on immunochromatography for the detection of H7 hemagglutinin (HA) antigen of influenza A virus was developed using 2 monoclonal antibodies that recognize different epitopes on the H7 HAs. The kit detected each of the tested 15 H7 influenza virus strains and did not react with influenza A viruses of the other subtypes than H7 or other avian viral and bacterial pathogens. The kit detected H7 HA antigen in the swabs and tissue homogenates of the chickens experimentally infected with HPAIV strain A/chicken/Netherlands/2586/03 (H7N7). The results indicate that the present kit is specific and sensitive enough for the diagnosis of HPAI caused by H7 viruses, thus, recommended for the field application as a pen-site test kit.     

Water adsorption properties of bamboo in the longitudinal direction

Differences in the water adsorption properties of moso bamboo (Phyllostachys pubescens) were examined by analyzing the isotherms. Hygroscopicity decreased from the bottom to the top of the culm, and this tendency was marked above about 80% relative humidity. Results of alkali extraction and the analysis of bundle sheath distribution revealed that the distribution of hygroscopic saccharides, for example, hemicelluloses and less-hygroscopic vascular bundles, affects the hygroscopicity, which varies depending on the position of the internode.     

Habitat management of little terns in Japan’s highly developed landscape

The little tern (Sterna albifrons) is a species of shorebird that nests in colonies on sandy beaches and riverbanks with little vegetation cover. In Japan, the natural breeding habitats of the terns have decreased drastically, and the species is listed under the category “vulnerable” in the Red List of the Government of Japan. To reduce the effects of habitat loss on the terns, conservationists are attempting to create and manage artificial colony sites in highly developed landscapes, such as reclaimed lands located in Tokyo Bay, central Japan. We present the factors that contribute to breeding success for this species and the habitat characteristics related to nest-site selection in the artificial colony sites along Tokyo Bay. Our data show that around 3 ha of a building rooftop without vegetation cover could sustain more than 2000 little tern nests. We found a strong positive relationship between colony size and hatching success, observing that a colony size of more than 100 lowered predation rates of eggs. On sites built upon white crushed-concrete, nest densities were high and predation rates of eggs were low. We also found that feeding rates were affected by foraging habitats, of which the best types were the shoreline of sandy beaches and mud flats. The 40 km dispersal range of the breeding terns, inferred from distribution data, should be considered when establishing a network of the multiple colony sites.     

Molecular cloning, nucleotide sequence and characteristics of a xylanase gene (xynA) from Ruminococcus albus 7

A 2.6-kb DNA fragment encoding a xylanase gene ( xyn A) was cloned from the rumen hemicellulolytic bacterium Ruminococcus albus 7. The deduced primary structure of the protein (XynA) was divided into a signal peptide region and 3 domains. Domain A was identified as a family 11 (G) catalytic domain, but one amino acid residue was replaced by another in an active site signature 1 of family 11. Domain B is a stabilizing domain for the catalytic domains of families 10 and 11. Deletion of domain B reduced stability of the xylanase at high temperature and at high and low pH. Domain B may be useful for protein engineering of xylanase. Domain C has sequence similarity to deacetylases and NodB proteins.     

Sex ratios of nigorobuna Carassius auratus grandoculis reared in paddy fields with fluctuating temperatures during larval and juvenile growth stages

The sex ratios of the nigorobuna Carassius auratus grandoculis reared in paddy fields during the sex differentiation periods of larval and juvenile stages were examined, since the phenotypic expression of sex in the fish is thermolabile, so that sex determination is under the control of genetic factors and temperature. After 2 days from hatching, the larvae were introduced into five paddy fields at five different densities in late May. After 37 or 41 days from hatching the juveniles were collected when the water was drained from the paddy fields, and reared in laboratory tanks supplied with running freshwater pumped from the lake shore until the age of 144–164 days. The mean water temperatures in the paddy fields were 22.4–24.1 ºC, and the maximum water temperature during the day frequently exceeded 30 ºC during the supposed critical period of sex determination of the fish. The sex ratios in the five paddy fields did not show a male-bias. Instead, all fields produced either balanced or slightly female-bias sex ratios. These results suggest that temporary high and fluctuating temperatures in paddy fields with low mean temperatures have hardly any masculinization effects on genotypic females of C. auratus grandoculis during sex differentiation stages.     

Carrier rate of Factor XI deficiency in stunted Japanese black cattle

Blood examinations and genotyping of Factor XI (F11) were performed in growth retardation Japanese Black cattle and their dams. Genotyping of F11 revealed that the recessive homozygous and heterozygous genotype frequencies were 5.2% and 50.0% in the Claudin-16 (CL-16) deficiency group (n=58), 0% and 14.2% in the renal dysplasia group (n=7), 0% and 26.1% in the non-CL-16 deficiency nephritis group (n=23), 8.9% and 46.7% in the hypogenesis syndrome group (n=45), 6.2% and 25.0% in the neonatal weak calf syndrome group (n=32), 9.1% and 38.6% in the respective dams group (n=44), 0% and 23.1% in the normal cattle group (n=13), and 5.9% and 38.2% in total (n=222), respectively. These results showed that the carrier rate of F11 deficiency was high in Japanese Black cattle, and that the CL-16 deficiency, hypogenesis syndrome, neonatal weak calf syndrome, and dams groups had a large amount of recessive homozygous genotype than the other groups. No abnormal bleeding was observed clinically in the present study, and 4 of the recessive homozygous dams showed normal growth and parturition.     

Parentage assignment in hatchery population of brown sole <Emphasis Type="Italic">Pleuronectes herzensteini</Emphasis> by microsatellite DNA markers

ABSTRACT:   Five loci ( Phz2 , Phz6 , Phz7 , Phz12 , and Phz14 ) of microsatellite DNA markers developed in a previous study for parentage assignment in the hatchery population generated by mating among 61 broodstock fish (35 females and 26 males) in a spawning tank, were selected. After natural spawning in the same tank, larvae collected at three different times were categorized into early phase (EP), middle phase (MP), and late phase (LP) groups. In the parental broodstock, the mean number of alleles per locus was 21.8 and expected heterozygosity ( H _E ) was 0.813. In the progeny, the mean number of alleles per locus decreased to 11.6 (EP), 14.4 (MP), and 6.4 (LP) and H _E to 0.796 (EP), 0.833 (MP), and 0.681 (LP). Parental assignment determined eight dams and six sires as major parents for the EP group. In the MP group, 13 dams and ten sires genetically contributed to spawning, but only three dams and two sires were involved in LP group progeny. In the hatchery population produced from a limited number of parental fish such as the LP group, genetic variability was apparently decreased.     

Cryopreservation of male and female gonial cells by vitrification in the critically endangered cyprinid honmoroko <Emphasis Type="Italic">Gnathopogon caerulescens</Emphasis>

We investigated the feasibility of cryopreservation of spermatogonia and oogonia in the critically endangered cyprinid honmoroko Gnathopogon caerulescens using slow-cooling (freezing) and rapid-cooling (vitrification) methods. Initially, we examined the testicular cell toxicities and glass-forming properties of the five cryoprotectants: ethylene glycol (EG), glycerol (GC), dimethyl sulfoxide (DMSO), propylene glycol (PG), and 1,3-butylene glycol (BG), and we determined cryoprotectant concentrations that are suitable for freezing and vitrification solutions, respectively. Subsequently, we prepared the freezing solutions of EG, GC, DMSO, PG, and BG at 3, 2, 3, 2, and 2 M and vitrification solutions at 7, 6, 5, 5, and 4 M, respectively. Following the cryopreservation of the testicular cells mainly containing early-stage spermatogenic cells (e.g., spermatogonia and primary spermatocytes), cells were cultured for 7 days and immunochemically stained against germ cell marker protein Vasa. Areas occupied by Vasa-positive cells indicated that vitrification led to better survival of germ cells than the freezing method, and the best result was obtained with 5 M PG, about 50% recovery of germ cells following vitrification. In the case of ovarian cells containing oogonia and stage I, II, and IIIa oocytes, vitrification with 5 M DMSO resulted the best survival of oogonia, with equivalent cell numbers to those cultured without vitrification. The present data suggest that male and female gonial cells of the endangered species G. caerulescens can be efficiently cryopreserved using suitable cryoprotectants for spermatogonia and oogonia, respectively.