Investigation of genetic diversity and inbreeding in a Japanese native horse breed for suggestions on its conservation

Because native breeds can serve as genetic resources for adapting to environment changes, their conservation is important for future agroecosystems. Using pedigree analysis, we investigated genetic diversity and inbreeding in Japanese Hokkaido native horses, which have adapted to a cold climate and roughage diet. Genetic diversity was measured as the number of founders and the effective number of founders, ancestors and genomes. All metrics imply a decrease in genetic diversity. A comparison of these metrics suggested that pedigree bottlenecks contributed more than did random gene losses to the reduction of genetic diversity. Estimates of marginal contributions of ancestors suggest that the bottlenecks arose mainly because related stallions had been used for breeding. A tendency for an increase in inbreeding coefficients was observed. F‐statistics revealed that a small effective population size majorly contributed to this increase, although non‐random mating in particular regions also contributed. Because the bottlenecks are thought to have reduced the effective population size, our results imply that mitigation of bottlenecks is important for conservation. To this end, breeding should involve genetically diverse stallions. In addition, to prevent non‐random mating observed in particular regions, efforts should be made to plan mating with consideration of kinships.     

Dominant Characteristics Between <Emphasis Type="Italic">Microcystis aeruginosa</Emphasis> and <Emphasis Type="Italic">Cyclotella</Emphasis> Sp. Accompanying Dilution Process in Eutrophic Lake

Although dilution of lake water has been used for improvement of water quality and algal blooms control, it has not necessarily succeeded to suppress the blooms. We hypothesized that the disappearance of algal blooms by dilution could be explained by flow regime, nutrient concentrations, and their interaction. This study investigated the effects of daily renewal rate (d), nitrogen (N) and phosphorus (P) concentration, and their interaction on the domination between Microcystis aeruginosa and Cyclotella sp. through a monoxenic culture experiment. The simulation model as functions of the N:P mass ratio and dilution rate (D) (calculated from d) was constructed, and the dominant characteristics of both species were predicted based on the model using parameters obtained in a monoculture experiment and our previous study. Results of monoxenic culture experiment revealed that M. aeruginosa dominated in all conditions (d = 5 or 15%; N = 1.0 or 2.5 or 5.0 mg-N L^?1; P = 0.1 or 0.5 mg-P L^?1) and the predicted cell densities were substantially correspondent to experimental data. Under various N:P ratios and D values, characteristics of domination for each species were predicted, indicating that Cyclotella sp. tended to be dominant under high P concentrations (P ≥ 0.36 mg-P L^?1) when the N:P ratio was less than 7.0, and M. aeruginosa could not form algal blooms at the N:P ratio ≤ 7.0 (N ≤ 0.7 mg-N L^?1). It was also suggested that the dilution rate leading to the Cyclotella sp. domination required 0.20 day^?1 or higher regardless of the N:P ratios.

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Graphical Abstract ? M. aeruginosa and Cyclotella sp. could be a superior competitor in nutrient-limited and nutrient-rich conditions, respectively. ? The simulation model in this study indicated that the predicted cell density and nutrient concentration were substantially correspondent to experimental data. ? The model predicted that Cyclotella sp. tended to be dominant at the P ≥ 0.36 mg-P L^?1 when the N:P ratio was less than 7.0, and M. aeruginosa could not form algal blooms at the N:P ratio ≤ 7.0 (N ≤ 0.7 mg-N L^?1).

     

Simultaneous extraction and fractionation and thin layer chromatographic determination of 14 mycotoxins in grains.

A simple, systematic analytical method for multiple mycotoxins was developed for detecting 14 mycotoxins; aflatoxins B1, B2, G1, and G2, sterigmatocystin, T-2 toxin, diacetoxyscirpenol, neosolaniol, fusarenon X, zearalenone, ochratoxin A, citrinin, luteoskyrin, and rugulosin. These mycotoxins were extracted with 20% H2SO4-4% KCl-acetonitrile (2 + 20 + 178), defatted with isooctane, and transferred to chloroform. The chloroform extract was cleaned up by silica gel column chromatography; the first 10 toxins were eluted with chloroform-methanol (97 + 3) and the remaining 4 toxins with benzene-acetone-acetic acid (75 + 20 + 5). Each fraction was analyzed by thin layer chromatography for the final determination. The method has been applied to polished rice, rough rice, corn, wheat, and peanuts as an analytical screening procedure. The detection limits in these commodities ranged from 10.00 to 800.0 microgram/kg, depending on the mycotoxin, but all limits were superior to those obtained for the individual mycotoxins by using other methods.     

Changes in GnRH levels in the brain and pituitary gland during migrations of sockeye salmon and chum salmon

Levels of two moleculer types of gonadotropin-releasing hormone (GnRH), salmon GnRH (sGnRH) and chicken GnRH–II (cGnRH–II) in the various brain regions and pituitary gland of sockeye salmon (Oncorhynchus nerka) and chum salmon (O. keta) during smoltification and spawning migration, respectively, were measured using specific time-resolved fluoroimmunoassay (TR-FIA) systems. Changes in sGnRH levels in different brain regions tended to be specifically synchronized with serum thyroid hormone or pituitary gonadotropin (GTH) levels during smoltification and spawning migration, respectively. In contrast, cGnRH–II levels did not show such synchronized changes. SGnRH and cGnRH–II in various brain regions might have different roles during smoltification and spawning migration of salmonid fishes.     

Effects of biotechnologically processed soybean meals in a nonfishmeal diet on growth performance, bile acid status, and morphological condition of the distal intestine and liver of rainbow trout Oncorhynchus mykiss

This study evaluated four commercially available biotechnologically processed soybean meals (SBMs) that were included as the primary protein sources in a nonfishmeal diet for juvenile rainbow trout Oncorhynchus mykiss. The processed SBMs were Afcep SBM fermented with mixed bacteria predominantly Bacillus spp., DaBomb SBM fermented with Lactobacillus acidophilus, PepSoyGen SBM fermented with Aspergillus oryzae and Bacillus subtilis, and HP300 SBM treated enzymatically. Fishmeal (FM) and common heat-treated and hulled SBM were used in the reference diets. Sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) analysis revealed that the protein components of Afcep SBM appeared to have relatively small molecular sizes compared with the other SBM products. After a 10-week feeding trial, growth of fish fed the common SBM diet was depressed, and abnormal liver and distal intestinal morphologies and bile acid characteristics were observed. The Afcep SBM diet showed approximated growth performance and comparable physiological conditions relative to the FM diet, while only limited improvements were observed in fish fed the other processed SBMs. These findings suggest that Afcep SBM, with its smaller molecular mass proteins, is the most promising processed SBM in nonfishmeal diets for rainbow trout.     

Changes in the immunostaining intensities of follicle-stimulating hormone and luteinizing hormone during ovarian maturation in the female Japanese flounder

The role of gonadotropin (GTH) in the reproduction of the Japanese flounder, Paralichthys olivaceus, was studied by assessing the changes in the apparent activity of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) in the pituitary gland during gonadal maturation by immunohistochemical analyses. Corresponding changes in plasma levels of testosterone (T), estradiol-17β (E₂), and 17α,20β-dihydroxy-4–pregnen-3-one (DHP) were also studied. Reared fish at the early spawning to termination stages were sampled from May to August and wild fish at the previtellogenic to termination stages were caught at 3- to 4-week intervals between April and September offshore from the northern mainland of Japan by gill nets. The gonadosomatic index of the reared fish decreased from the early spawning stage to the termination stage, while that of the wild fish increased significantly from the previtellogenic stage to the early spawning stage and decreased thereafter. In the reared fish, the immunostaining intensities of FSH and LH were high during the spawning period, accompanied by high plasma levels of T, E₂, and DHP. In the wild fish, the immunostaining intensities of FSH and LH were low during the previtellogenic stage but increased during the maturing and spawning stages. These results indicate that both FSH and LH are likely associated with oocyte maturation in the Japanese flounder.     

Tetrodotoxin accumulation conflicts with low salinity tolerance in juvenile tiger puffer Takifugu rubripes

We evaluated whether bearing tetrodotoxin (TTX) affects salinity stress in the juvenile tiger puffer Takifugu rubripes. Juveniles of hatchery-reared non-toxic T. rubripes [body weight (BW): 1.7?±?0.2 g, n?=?120] were divided into six tanks and acclimatized to salinity (8.5 ppt) that is equivalent to blood osmolality. Fish in three tanks were fed non-toxic diet, and those in the other three tanks were fed a TTX-containing diet (356 ng/g diet) three times a day until satiation. In each diet treatment, salinity of one tank was kept at 8.5 ppt, and the other two tanks were adjusted to either 1.7 or 34.0 ppt, and fish were reared for another 33 days. Then, we compared survival, growth, TTX accumulation, plasma osmolality, plasma cortisol, and glucose levels among treatments. We detected TTX only in the fish in the TTX-diet groups. Survival was highest at 8.5 ppt (70%) and lowest at 1.7 ppt in the TTX-diet group (20%). The BW was greater at 8.5 ppt, and plasma osmolality was significantly higher at 34.0 ppt than at any other salinities. Plasma cortisol level was significantly higher but glucose level was lower at 1.7 ppt. Possessing TTX at a low salinity may be lethal to tiger puffer juveniles.

     

Evidence for existence of cAMP-Epac signaling in the heads of mouse epididymal spermatozoa

Cyclic adenosine 3',5'-monophosphate (cAMP) signaling regulates the expression of fertilizing ability in mammalian spermatozoa. Many articles indicate that this signaling is mediated mainly via protein kinase A. Recently, a guanine nucleotide exchange factor for small G protein Rap1 (an exchange protein directly activated by cAMP: Epac) was discovered as a new mediator of cAMP signaling in somatic cells. The aim of this study was to reveal the existence of cAMP-Epac signaling in mouse spermatozoa. Northern blot analysis and in situ hybridization suggested that Epac1 and Epac2 mRNAs were transcribed in the seminiferous epithelia of the testis. This shows that expression of Epac mRNAs is present in mouse testicular germ cells. Indirect immunofluorescence with specific polyclonal antibodies suggested possible co-localization of Epac1 and Rap1 proteins in the heads of epididymal spermatozoa. Moreover, treatment of epididymal spermatozoa with an Epac-specific cAMP analog, 8-pMeOPT-2'-O-Me-cAMP, induced activation of Rap1, as revealed with a commercial kit for pull-down assay. These results indicate the existence of cAMP-Epac signaling in the heads of mouse epididymal spermatozoa.     

Evaluation from otolith Sr stable isotope ratios of possible juvenile growth areas of Japanese eels collected from the West Mariana Ridge spawning area

Fisheries Science - The widely distributed East Asian Japanese eel Anguilla japonica constitutes a single genetically homogeneous population with a single spawning area near the West Mariana Ridge....     

人工合成小麦根愈伤组织细胞和不定根的染色体变异

本研究以日本京都大学农学部生殖质研究所人工合成小麦品系和普通小麦品种“中国春”为材料,种子根为外植体诱导出愈伤组织和不定根,对其染色体数目的变异进行了分析比较。结果表明,品系间愈伤组织细胞的染色体变异有很大差别,品系89W36的变异幅度较大2n=7～58,品种“中国春”（88W70）的染色体变异较小2n=15～43。 小麦染色体组基数 X=7的整倍体 14、21、28、35、42的变异频率都大于相近数目的非整倍体。小麦正常染色体数42的频率比其他数目的变异高。不定根染色体的变异与愈伤组织相比相对稳定,最大变异范围是2n=25-45,染色体数的变异主要集中在39～44,各品系正常染色体数42的细胞频率都在68％以上。