Production of inhibin A and inhibin B in boars: changes in testicular and circulating levels of dimeric inhibins and characterization of inhibin forms during testis growth

We investigated the production of inhibin in boars from the infantile to pubertal periods by: (1) measurement of testicular and circulating levels of inhibin, (2) characterization of inhibin forms and (3) localization of inhibin subunits in the testis. Total inhibin levels in the testis increased until 8 weeks of age but then declined to much lower values at 15 weeks. Testicular inhibin A and inhibin B were high until 8 weeks. Circulating levels of total inhibin and inhibin A were also high until 8 weeks, then declined from 10 weeks; inhibin B was not detected, because of low sensitivity of the inhibin B assay. Analyses of inhibin A and inhibin B levels in the eluted fractions obtained from testes after immunoaffinity chromatography and SDS-PAGE showed the presence of a peak of approximately 45 kDa until 10 weeks of age. As the boars aged, the levels of inhibin A and inhibin B increased in the molecular weight region of 29–31 kDa. The fractions corresponding to 29 and 30 kDa suppressed FSH release from rat pituitary cells, but the 45 kDa fraction had no FSH-suppressing activity. Total amounts of inhibin A isolated from the SDS gels were similar to those of inhibin B until 10 weeks of age, but were three times higher than those of inhibin B between 15 and 25 weeks. Further fractionation by reverse phase high-performance liquid chromatography revealed that the 29–31 kDa immunoreactive material was composed of mature forms of inhibin A and inhibin B, in addition to a 26 kDa monomer. Immunohistochemistry indicated that positive immunostaining for the subunits was observed in Sertoli cells from the infantile to pubertal periods. Elongated spermatids also showed positive signals at age 25 weeks. These results clearly indicated that: (1) the boar testis has the ability to produce inhibin A and inhibin B during the infantile period but inhibin A is the predominant form towards puberty and (2) the molecular weight forms of inhibin and the sites of production of inhibin change with testicular development.     

Effects of dietary astaxanthin and vitamin E and their interactions on the growth performance,pigmentation, digestive enzyme activity of kuruma shrimp (Marsupenaeus japonicus)

A 56‐day feeding trial was done to investigate the interactive effects of astaxanthin (Ax) and vitamin E (α‐Toc) on the performance of kuruma shrimp (Marsupenaeus japonicus). A 2 × 3 factorial experiment was conducted with six experimental diets containing two levels of Ax (0 and 0.6 g/kg diet [Ax₀ and Ax_0.6]) and three levels of α‐Toc (0, 0.2 and 1 g/kg diet [α‐Toc₀, α‐Toc_0.2 and α‐Toc₁]). The results indicated that growth performance was significantly (p < 0.05) increased in shrimp fed with the Ax_0.6 × α‐Toc_0.2 diet. Interactive effects between Ax and α‐Toc on the growth parameters were observed. Furthermore, pigmentation performance was significantly (p < 0.05) better in the Ax_0.6 groups. Interaction between Ax and α‐Toc was also found in the Ax content of shrimp body parts. Interestingly, dietary α‐Toc helped to reduce the Ax consumption rate, promote the absorption and increase the deposition of Ax in the muscle. Shrimps from the Ax_0.6 groups showed significantly (p < 0.05) improved hepatopancreatic digestive enzyme activities compared with those of the Ax₀ groups. Although no interactive (p > 0.05) effects were found between dietary α‐Toc and Ax on total haemocyte count and tolerance against freshwater, dietary Ax and α‐Toc supplementation showed better performance on these two parameters. It was concluded that dietary Ax and α‐Toc functioned synergistically, and the shrimp fed with the diet containing 0.6 g Ax/kg diet Ax and 0.2 g α‐Toc/kg diet showed improved growth and pigmentation performance compared with the other groups in the current study.     

Inhibition of Df-protease associated with allergic diseases by polyphenol.

It was reported that Df-protease from house dust mite (Dermatophagoides farinae) catalyzes the activation of the kallikrein-kinin system in human plasma and is closely associated with mite-induced allergy. Therefore, to prevent the release of kinin by Df-protease, the inhibitory activity of polyphenols including catechins and flavonols was tested in vitro and in vivo. Among them, myricetin and epigallocatechin gallate (EGCg) effectively inhibited the amidase activity of Df-protease with K(i) values of 1 x 10(-)(8) and 6 x 10(-)(4) M, respectively. The kinin release in human plasma was extensively inhibited by the addition of EGCg in comparison with myricetin. Enhancement of vascular permeability in guinea pigs caused by Df-protease was markedly suppressed by EGCg.     

Preparation of Anti-Rabies Virus N Protein IgYs by DNA Immunization of Hens Using Different Types of Adjuvants

DNA immunization has been used to study vaccination methods and for production of specific antibodies. The present study aimed to apply DNA immunization to prepare specific IgYs, which react against rabies virus N protein (RV-N) and can be used to research and diagnose rabies virus. The DNA sequence of RV-N was ligated into a pcDNA 3.1 plasmid for constructing pcDNA-N. Eight hens were divided into four groups. Group 1 comprised the control group (non-immunized). In Groups 2, 3, and 4, hens were injected intramuscularly with pcDNA-N (400 µg/hen). Eight injections were administered every other week. From the 4th week, an adjuvant was injected in addition to pcDNA-N. Freund''s complete adjuvant (FCA) and λ-carrageenan were administered to Groups 3 and 4, respectively. Eggs were collected daily, and the specific antibody activities of egg yolks were measured by ELISA. IgYs were purified from pooled egg yolks at 16–19 weeks post-administration in each group. The detection sensitivities of the RV-N were compared using purified IgY as the primary antibody for ELISA, dot blotting, and western blotting. Egg yolks from one of the two hens in Group 2 (pcDNA-N alone) and all hens in Groups 3 (pcDNA-N + FCA) and 4 (pcDNA-N + λCarra) had increased ELISA values. The combined use of λ-carrageen in DNA immunization resulted in an adjuvant effect comparable to that of FCA. Each purified specific IgY detected RV-N in the ELISA, western blotting, and dot blotting; however, the detection sensitivity differed. Higher detection sensitivity of the +λCarra IgY was observed by ELISA, whereas there was higher detection sensitivity of +FCA IgY in western blotting and dot blotting. In summary, anti-rabies virus N protein IgY was prepared through DNA immunization of hens using FCA or λ-carrageenan as adjuvants and can be used as a primary antibody to detect rabies viruses.     

Springiness of Pancake and Its Relation to Binding of Prime Starch to Tailings in Stored Wheat Flour

Wheat flours were stored at room temperature (15–25°C), 40, 60, 80, and 100°C for various times. The baking performance of these flours was then evaluated in terms of the springiness of pancakes (recovery from crushing). Baking performance improved with increased storage time at each temperature. Brabender Amylograph tests of the flours indicated that the onset temperature in viscosity decreased with increased storage time at each temperature. When the flours were fractionated by acetic acid (pH 3.5) with mortar and pestle, recoveries of the water-solubles and gluten fractions were unchanged, but recoveries of prime starch and tailings fractions changed remarkably with increased storage time. On the other hand, those changes were not observed when flours were fractionated with a Waring blender. The binding of prime starch to tailings was correlated significantly with baking performance.     

Multiplex PCR method for differentiating highly pathogenic Yersinia enterocolitica and low pathogenic Yersinia enterocolitica,and Yersinia pseudotuberculosis

A multiplex PCR method for rapid and sensitive diagnosis, differentiating three pathogenic Yersinia groups such as the highly pathogenic Y. enterocolitica, including serotype O8, low pathogenic Y. enterocolitica, and Y. pseudotuberculosis, was developed. Four primer pairs were chosen to detect the genes fyuA, ail, inv, and virF, responsible for the virulence in pathogenic Yersinia species. Under the multiplex PCR conditions, the unique band patterns for the highly pathogenic Y. enterocolitica, low pathogenic Y. enterocolitica, and Y. pseudotuberculosis were generated from Yersinia strains. The detection limit of this method was 10¹–10³ CFU per reaction tube. This multiplex PCR method could detect highly pathogenic Y. enterocolitica O8 from the wild rodent fecal samples that were culture-positive. Therefore, the new multiplex PCR method developed in this study is a useful tool for rapid and sensitive diagnosis, distinguishing three pathogenic Yersinia groups.     

Understanding why farmers plant trees in the homestead agroforestry in Bangladesh

Because trees cover only 6.4% of the total land area of Bangladesh, while agricultural expansion continues to massively deplete the natural forests, a well-managed homestead forestry practice is vital for reversing the existing trend and promoting the ecological balance of the country. An understanding of the decision-making process of the farmers who practice homestead forestry is important in expanding and improving the practice. This paper seeks to characterize and analyze factors influencing farmers' decisions about tree planting. Logistic and multiple regression analyses were applied to determine the factors that influence the farmers' tree-planting decisions. The analyses demonstrate a number of important conclusions: (i) tree-planting increases with the amount of homestead land owned; (ii) farmers whose main source of income is non-agricultural are more likely to decide to plant trees in the homestead; (iii) purchasing cost of fuelwood has a positive influence on tree-planting decisions; (iv) number of male family member has a positive influence on farmers' tree-planting decisions; and (v) knowledge of the activities of the forestry extension programs has a positive influence on tree-planting decisions. The results of the study demonstrate that, in recent years, farmers' decisions of whether or not to plant trees have been based primarily on economic rather than ecological concerns. It is concluded that there is substantial potential for the improvement of homestead forestry, and that properly managed homestead forestry can alleviate the poverty of rural people by increasing overall household income. To this end, it is suggested that forestry extension workers work more closely with the local people in order to implement homestead forestry.This revised version was published online in November 2005 with corrections to the Cover Date.     

Individual-scale responses of five dominant tree species to single-tree selection harvesting in a mixed forest in Hokkaido, northern Japan

We investigated the individual-scale responses of five dominant species (Abies sachalinensis, Acer mono, Tilia japonica, Quercus crispula, and Betula ermanii) to single-tree selection harvesting in a conifer–broadleaved mixed forest in Hokkaido, northern Japan. Using data from stems with a diameter at breast height of ≥12.5 cm, collected during 20 years of monitoring a 6.7-ha stand, we analyzed the effects of harvesting in the neighborhood on tree recruitment and the growth (diameter class transition) and mortality of the residual trees. The effects of harvesting varied considerably among tree sizes and species. Harvesting improved the recruitment of A. mono and B. ermanii, and moderated the negative effect of the initial basal area of the surrounding canopy trees on the recruitment of Q. crispula. Conversely, harvesting limited the recruitment of A. sachalinensis by offsetting the positive effect of the initial basal area of the surrounding canopy trees. The growth of A. sachalinensis and Q. crispula decreased with the initial basal area of the surrounding canopy trees. Harvesting in the neighborhood resulted in an improvement in the growth of the trees of these species only in the smaller size classes. With increasing local harvesting intensity, the mortality of smaller A. sachalinensis trees decreased, whereas the mortality of larger trees increased. These results suggest that differences in the local harvesting intensity, spatial patterns of harvesting, and initial stand structures influence the stand-scale dynamics in response to partial harvesting in the mixed forests of this region.     

Individual growing conditions that affect diameter increment of tree saplings after selection harvesting in a mixed forest in northern Japan

We analyzed temporal patterns in diameter growth of saplings following selection harvesting in an uneven-aged mixed stand dominated by Abies sachalinensis, Acer mono, Quercus crispula, and Betula ermanii in Hokkaido, northern Japan. We examined interspecific differences in growth responses to local growing conditions including harvesting intensity, crowding, stem size, and past duration of the small growth period. Consistent with expectations based on shade tolerance of the species, the age at which the individual reached a diameter at breast height (DBH) of 12.5 cm was highest for A. sachalinensis and lowest for B. ermanii. The interspecific growth differences between saplings that had or had not experienced local harvesting increased gradually for A. sachalinensis and B. ermanii, but peaked at around 4–6 years after harvesting for Q. crispula. Generalized linear mixed model analysis clearly suggested that individual growth conditions required to enhance diameter growth of saplings differed considerably among species. For Q. crispula and B. ermanii, local harvesting intensity was most strongly and positively associated with diameter growth rate, whereas for A. sachalinensis and A. mono, stem size had the strongest negative effect. Abies sachalinensis saplings responded more to surrounding harvesting when they were relatively small, whereas A. mono showed a weak opposite response. The duration of the small growth period before harvesting had negative effect for A. sachalinensis, but not for the other species. Our study indicated that the influence of selection harvesting on growth of shade-tolerant species depends upon pre- and post-harvest growing conditions.     

Effects of water-soluble and fat-soluble choline sources on the performances of juvenile kuruma shrimp, Marsupenaeus japonicus Bate

The present study was undertaken to clarify the effect and interaction of choline chloride (CC) and phosphatidylcholine (PC), the water‐ and fat‐soluble sources of choline, on the performances of kuruma shrimp, Marsupenaeus japonicus juveniles using as response variables such as percent weight gain (%WG), specific growth rate (SGR), survival (SR%), feed efficiency ratio (FER), protein efficiency ratio (PER), proximate composition, fatty acids profile and free choline and PC contents in the whole body of kuruma shrimp. A 40‐day feeding trial was conducted as a 2 × 3 factorial experiment with six test diets containing three levels of CC (0.3, 0.6 and 1.2 g kg^?1) and two levels of soybean PC (SPC; 2 and 4 g kg^?1). Both of the choline sources (CC and SPC) showed a significant interaction (P<0.05) regarding all the evaluated response variables except for the proximate and fatty acid contents. Interestingly, there was a positive effect (P<0.05) of CC observed with increasing its level at the low level of SPC and vice versa. However, the magnitudes of the beneficial effect of CC in terms of the evaluated parameters were changed with the different dietary levels of SPC. Also, the inclusion of higher levels of SPC (4 g kg^?1) or CC (0.6 and 1.2 g kg^?1) diminished the effect of the other source.