Sex determination of sika deer (Cervus nippon yesoensis) using nested PCR from feces collected in the field

We describe a method for determining the sex of sika deer (Cervus nippon yesoensis) from feces collected in the field. Using a nested polymerase chain reaction (nested PCR), partial sequences of the sex determination region of the Y chromosome (SRY) gene and X zinc finger protein (ZFX) gene were amplified. In 19 individuals with sex information, the correct sex was successfully detected and sequences of target amplicons were completely matched between muscle and feces from the rectum. Among 75 fecal samples collected noninvasively in the field, 68-71 samples (90.7-94.7%) yielded successful sex determinations. Using this technique, feces collected in the field would enhance the utility of genetic analysis. As a result, instead of biomaterials, these samples can serve as new or alternative materials. Finally, it can be expected that this technique will contribute to reveal in advanced detail of the population dynamics and genetic diversity that needed to carry out effective population control and to reduce the extinction risk of sika deer.     

Quantitative analyses of lymphoid tissue in the spleen, lymph nodes and Peyer's patches in cynomolgus monkeys

To clarify the morphological characteristics of the cynomolgus monkey immune system, we analyzed quantitative data on their lymphoid organs. Spleens, major lymph nodes and Peyer's patches were sampled from cynomolgus monkeys, and the lymphoid follicle and germinal center areas and percentages of CD3- and CD20-positive areas were calculated. All the organs analyzed showed large interindividual variations in the sizes of lymphoid follicles and germinal centers. Lymphoid follicle in the spleen, submandibular lymph nodes and Peyer's patches showed no marked difference in size. Germinal center size in the mesenteric lymph nodes and Peyer's patches were significantly smaller than those in the spleen. Areas containing T cells were largest in the lymph nodes, while those containing B cells were largest in the spleen and Peyer's patches. The mean size of the splenic lymphoid follicle in cynomolgus monkeys is larger than that in rats and similar to that in humans. Based on the large individual variation and the characteristics of lymphoid organs, it is important to use cynomolgus monkeys in standard toxicity studies. Taking advantage of the characteristics of each species enables reliable evaluation of the immunologic system in standard toxicity studies.     

Role of neck region in the thermal aggregation of myosin.

Carp dorsal myosin formed oligomers that retained ATPase activity upon heating. Cleavage of the oligomeric myosin at subfragment-1 (S-1)/rod junction released monomeric S-1 and rod, indicating that ATPase retaining myosin associated near the S-1/rod junction. The digest also contained rod oligomers. Heating a mixture of S-1 and rod generated neither ATPase retaining S-1 oligomers nor rod oligomers. Electron microscopic observation of the heated myosin revealed that some oligomers were formed by associating at the S-1/rod joining region, exhibiting a recognized double head, probably ATPase retaining oligomers. No myosin oligomers associated at the tail region were observed, thus, rod aggregation would be formed at its very restricted region near the S-1/rod junction. Based on the findings, we proposed that the neck structure is important in the thermal oligomerization process of myosin.     

Height growth,diameter-height relationships and branching architecture of <Emphasis Type="Italic">Pinus massoniana</Emphasis> and <Emphasis Type="Italic">Cunninghamia lanceolata</Emphasis> in early regeneration stages in Anhui Province,eastern China: effects of light intensity and regeneration mode

To determine the growth strategies during early regeneration stages of two major coniferous species (Pinus massoniana     

Protein metabolism in rice seedling

The relationship between protein synthesis and degradation in germinating rice seed were studied with protein synthetic inhibitors. Both DNP and 8-AG inhibited the degradation of glutelin, the major storage protein in rice seed, while the inhibitors had no direct effect in the activity of rice seed proteinllse in vitro. The prevention can be partly ascribed to the Inhibition of proteinase synthesis because the inhibitors depressed the increase in proteinase activity during germination. When DNP treatment was started at the onset of germination, the degradation of glutelin in the endosperm was seriously inhibited and the endosperm remained rigid over 9 days of incubation at 30°C. In contrast, the inhibition was less efficient clent when the treatment was started in the later stage. It is suggested that the degradation of the storage protein in rice seed depends on the synthetic process of the hydrolytic enzymes which increase during germination. disintegrate the compartmentation of the endosperm and allow the storage proteins to come in contact with the existing proteinases     

Studies on the effect of the paddy field soil kept under upland condition on the yield of rice

In Japan, the extent under paddy is approximately three million hectares and the average yield per hectare is about 3000 kg, though high yield gives as much as over 7000 kg per hectare. It is at present a serious problem as to what methods we can use to obtain a high yield of rice. Generally, in well-drained paddy fields where the system of double-cropping is practised, rice is cultivated under a flooded condition in summer, and upland crops such as barley or wheat is cultivated under upland condition in other seasons.     

Supplemental effect of bile salts to soybean meal-based diet on growth and feed utilization of rainbow trout <Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>

A feeding experiment was conducted to evaluate the supplemental effect of bile salts to a defatted soybean meal-based non-fish meal diet for rainbow trout Oncorhynchus mykiss. A fish meal-based diet (FM) and two non-fish meal diets with and without bovine bile salts (SC+B and SC, respectively) were fed to fish (13 g initial weight) for 10 weeks. Fish fed diet SC showed inferior growth and feed efficiency, while bile salt supplementation improved the parameters to the same levels as fish fed diet FM. Crude fat and starch digestibility of diet SC-fed fish decreased after the 10-week feeding trial compared to the data obtained with fish that had no experience of the diet. Total biliary bile salt content and intestinal maltase activity of fish fed diet SC were the lowest among treatments, while these parameters were improved by bile salt supplementation. Morphological changes occurred in the distal intestine and liver of the diet SC group, although the histological features of fish fed diet SC+B were similar to those of fish fed diet FM. These results suggest that bile salt supplementation to a soybean meal-based diet improves the nutrient utilization by normalizing digestive processes in rainbow trout.     

Response of enzyme activities and metabolic intermediate concentrations to epinephrine administration in hepatopancreas and muscle of carp

SUMMARY: Common carp Cyprinus carpio were administered intraperitoneally with 10 μmol epinephrine per 100 g bodyweight, and enzyme activities and metabolic intermediate concentrations were determined in the hepatopancreas and muscle. Glycogen phosphorylase a (GPase a) activity together with cyclic AMP (cAMP) concentration was increased, and glycogen content was decreased in the hepatopancreas and muscle at 2 h after the administration. The epinephrine administration also increased hepatopancreatic glucose-6-phosphatase and fructose-1,6-biphosphatase activities as well as serum glucose, lactate, and free amino acid concentrations. Furthermore, its administration increased phosphofructokinase activity together with lactate, fructose-6-phosphate, adenosine-5'-monophosphate, and adenosine-5'-diphosphate concentrations and decreased citrate concentration in the muscle. Thereafter, almost all parameter concentrations in the hepatopancreas and muscle recovered to the pre-administered levels during 24 h after the administration. These results suggest that epinephrine administration enhanced glycogenolysis and gluconeogenesis in the hepatopancreas, and released glucose into the bloodstream to supply it to the muscle. The blood glucose together with muscle glycogen seems to be metabolized through enhanced glycolysis in the muscle.     

Purification of serine carboxypeptidase from the hepatopancreas of Japanese common squid Todarodes pacificus and its application for elimination of bitterness from bitter peptides

ABSTRACT:   An acidic serine carboxypeptidase (CPase Tpa) from the hepatopancreas of Japanese common squid Todarodes pacificus was purified. Purified CPase Tpa had a molecular mass of 36 kDa on sodium dodecylsulfate–polyacrylamide gel electrophoresis, and an isoelectric point of 6.0. The optimum pH of CPase Tpa was pH 4.0. In investigating the specificity of CPase Tpa for several peptide substances, it was found that peptides with hydrophobic or bulky amino acid residues at the P₁ position reacted well. The enzymatic activity was almost completely inhibited by p -chloromercuribenzoic acid, monoiodoacetic acid, diisopropylfluorophosphate and HgCl₂. This is the basis for its grouping in the serine carboxypeptidase family (EC 3. 4. 16. 5). The substrate specificity of CPase Tpa can be used to eliminate the bitterness of bitter peptides. In this study, the bitterness-reductive effect using bitter peptides prepared by hydrolyzing soy protein, casein and corn gluten with pepsin or trypsin was tested. The bitterness of soy peptide digested with pepsin was completely eliminated by treatment with CPase Tpa, whereas the bitterness of casein digested with trypsin and corn peptide digested with pepsin were somewhat less efficient. On the basis of these results, it is anticipated that CPase Tpa would be effective in eliminating the bitterness of some bitter peptides.     

Molecular Detection of Avian Pathogens in Poultry Red Mite (Dermanyssus gallinae) Collected in Chicken Farms

Poultry red mite (PRM, Dermanyssus gallinae) is a blood-sucking ectoparasite as well as a possible vector of several avian pathogens. In this study, to define the role of PRM in the prevalence of avian infectious agents, we used polymerase chain reaction (PCR) to check for the presence of seven pathogens: Avipox virus (APV), Fowl Adenovirus (FAdV), Marek’s disease virus (MDV), Erysipelothrix rhusiopathiae (ER), Salmonella enterica (SE), Mycoplasma synoviae (MS) and Mycoplasma gallisepticum (MG). A total of 159 PRM samples collected between 2004 and 2012 from 142 chicken farms in 38 prefectures in Japan were examined. APV DNA was detected in 22 samples (13.8%), 19 of which were wild-type APV. 16S ribosomal RNA (16S rRNA) of MS was detected in 15 samples (9.4%), and the mgc2 gene of MG was detected in 2 samples (1.3%). Eight of 15 MS 16S rRNA sequences differed from the vaccine sequence, indicating they were wild-type strains, while both of the MG mgc2 gene sequences detected were identical to the vaccine sequences. Of these avian pathogen-positive mite samples, three were positive for both wild-types of APV and MS. On the other hand, the DNAs of ER, SE, FAdV and MDV were not detected in any samples. These findings indicated that PRM can harbor the wild-type pathogens and might play a role as a vector in spreading these diseases in farms.