Effects of synthetic kisspeptin peptides and GnRH analogue on oocyte growth and circulating sex steroids in prepubertal female chub mackerel (Scomber japonicus)

In recent years, kisspeptin peptides, encoded by kiss genes have been used to manipulate reproductive processes in farmed animals, including fish. Our previous studies demonstrated that the chub mackerel brain expresses kiss1 and kiss2 and intramuscular injection of synthetic Kiss1 pentadecapeptide (Kiss1‐15) but not Kiss2 dodecapeptide (Kiss2‐12) accelerates spermatogenesis in prepubertal male chub mackerel (Scomber japonicus). In the present study, we evaluated their effects in prepubertal female chub mackerel. The gonadosomatic index (GSI) values of experimental fish did not show any significant changes. Condition factor (CF) values increased significantly in Kiss1‐15 treated fish, in comparison with control and GnRH analogue (GnRHa) injected fish. Histologically, only perinucleolar oocytes were found in all experimental fish. However, Kiss and GnRHa treated fish showed a significant increase in the perinucleolar oocyte diameter, in comparison with the control fish. Gene expression analyses revealed decreased expression of gnrh1 in the telencephalon‐preoptic region of the brain of Kiss2‐12 and GnRHa injected fish, in comparison with control fish. In contrast, GnRHa injected fish exhibited higher levels of fshβ in the pituitary, with no changes in the levels of lhβ among different treatments. Levels of circulating sex steroids, testosterone, and estradiol‐17β were significantly higher in Kiss1‐15 injected fish, in comparison with control fish. These results indicate that synthetic kisspeptin peptides and GnRHa can induce oocyte growth in prepubertal female chub mackerel.     

Analysis of genetic and biological characters of Japanese potato strains of <Emphasis Type="Italic">Ralstonia solanacearum</Emphasis>

We assessed the geographic distribution, biovar, phylotype, DNA fingerprints (rep-PCR), and/or endoglucanase sequence of potato bacterial wilt pathogen, Ralstonia solanacearum (Rs), in Japan. Rs has been isolated from potato fields in southwestern, warm, temperate regions. Of the 188 isolates, 74 belonged to biovar N2 (39%), 44 to biovar 3 (24%), and 70 to biovar 4 (37%). Biovars N2 and 4 strains were widely distributed, from northern (Hokkaido) to southern (Okinawa) Japan. Based on the results of multiplex-PCR analysis, every potato strains belonged to either phylotype I or IV. Phylotype I comprised both biovars 3 and 4 strains. On the other hand, phylotype IV included biovar N2 strains. None of the strains belonged to phylotype II or III or biovar 1 or 2. Phylogenetic analysis based on DNA fingerprints and endoglucanase gene sequences clarified the genetic diversity of the Japanese potato strains and the close genetic relationship between the Japanese strains and the Asian strains in phylotypes I and IV.     

Antiobese effects of novel saponins from edible seeds of Japanese horse chestnut (Aesculus turbinata BLUME) after treatment with wood ashes

Recently, we have identified novel saponins from edible seeds of Japanese horse chestnut ( Aesculus turbinata BLUME) after processing the natural seeds with wood ashes to remove bitterness. We attempted to determine anti-obesity effects of those saponins from edible seeds as well as natural seeds. The purified individual components of saponins from natural and edible seeds inhibited pancreatic lipase in vitro. The potency was in the order of escins > desacylescins > deacetylescins. Escins Ib and IIb as well as deacetylescins Ib and IIb with the angeloyl moiety were more potent than the corresponding Ia and IIa series with the tigloyl moiety. Moreover, in vivo anti-obesity effects of the saponin fractions were monitored for 8 weeks in mice fed high-fat diets. Saponin fractions from both seeds significantly attenuated the elevation in body weight, the mass of peritoneal adipose tissues, and plasma triacylglycerol, which was accompanied by higher contents of undigested fats in feces without changes in food intake, indicating the effective inhibition of fat digestion in vivo. Taken together, saponin fractions including desacylescins and deacetylescins from edible seeds are potentially useful for the development of nutraceutical foods with anti-obesity effects and more attenuated bitter taste.     

Factors Affecting Gestation Length in Heavy Draft Mares

The purpose of this study was to examine the factors affecting gestation length in 209 foalings from 65 heavy draft horses by multiple linear regression analysis. The objective variable was gestation length, and the explanatory variables were stud farm, year foaled, month foaled, age of mare, parity of mare, last gestation length, foal gender, and some meteorological parameters (the mean values of prepartum 10 days: maximum temperature, minimum temperature, average barometric pressure, and hours of sunshine in a day). The mean gestation length for 209 foalings was 334.9 (standard deviation = 8.3) days, with a range of 313-352 days. Stepwise regression analysis showed that last gestation length and foal gender had significant (P < .01) effects on gestation length. Multiple regression analysis showed that last gestation length (standard partial regression coefficient = 0.54) was more influential on the present gestation length than foal gender (standard partial regression coefficient = 0.22). The present gestation length and the last gestation length were moderately positively correlated (r = 0.56, P < .01). Colt foal (n = 106) pregnancies were significantly (P < .01) longer than fillies (n = 103) (mean ± SEM: colt 337.1 ± 0.7 vs. filly 332.7 ± 0.9). According to our results, last gestation length (mare itself) and foal gender were the main factors affecting gestation length in heavy draft horses. Once several gestation lengths are known, predictions with acceptable accuracy can be made about the gestation length of that mare's following foaling.     

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.     

Allele frequency distribution of the canine dopamine receptor D4 gene exon III and I in 23 breeds

Various canine breeds are remarkably different from each other not only in their sizes and shapes but also in behavioral traits, suggesting that some of them are under genetic control. Although dopaminergic neurotransmission system is considered to affect animal behavior, little is known about related genes in canine. Relations between specific alleles in polymorphic regions of the dopamine receptor D4 gene (DRD4) and personality or psychiatric disorders have been reported in humans, and we first found polymorphism in exon III region of the gene in 4 canine breeds. In this study we surveyed allele frequency distribution in 23 breeds including a total of 1,535 unrelated individuals. In exon III, 8 alleles including a novel allele were identified. A group of breeds in which the alleles 447b, 498 and 549 were frequent tended toward high scores in aggression-related behavioral traits than that with frequent alleles 435 and 447a. Moreover, a polymorphism based on 24 bp insertion/deletion was found in exon I region for the first time in dogs. This information may be of use for candidate gene studies of behavioral variation in dogs.     

Seedling rot of garland chrysanthemum caused by Gibellulopsis chrysanthemi and ecological characters of the causal fungus

A new disease found on garland chrysanthemum in Osaka, Japan in 2009 and its causal agent were identified and characterized. Light brown spots first appeared on lower leaves of seedlings, and the leaves blighted or rotted. A fungus isolated from diseased plants, described recently as Gibellulopsis chrysanthemi, was demonstrated to reproduce the natural symptoms in inoculation tests. Sufficiently long, moist periods after inoculation promoted the infection even after lengthy dry periods. The pathogen also caused the disease on chrysanthemum and lettuce, but not on seven other vegetables. The fungus caused the disease after 6 months of dry storage. The disease was termed seedling rot (“nae-fuhai-byo” in Japanese).     

Pathogenicity evaluation of Ralstonia pseudosolanacearum race 4 on ginger by leaf-clipping inoculation

Journal of General Plant Pathology - An easy, rapid method has been needed to test the pathogenicity of strains of Ralstonia pseudosolanacearum, the cause of serious bacterial wilt on ginger...     

Changes in the expression of pituitary gonadotropin subunits during reproductive cycle of multiple spawning female chub mackerel Scomber japonicus

The endocrine regulation of reproduction in a multiple spawning fish with an asynchronous-type ovary remains largely unknown. The objectives of this study were to monitor changes in the mRNA expression of three gonadotropin (GtH) subunits (GPα, FSHβ, and LHβ) during the reproductive cycle of the female chub mackerel Scomber japonicus. Cloning and subsequent sequence analysis revealed that the cDNAs of chub mackerel GPα, FSHβ, and LHβ were 658, 535, and 599 nucleotides in length and encoded 117, 115, and 147 amino acids, respectively. We applied a quantitative real-time PCR assay to quantify the mRNA expression levels of these GtH subunits. During the seasonal reproductive cycle, FSHβ mRNA levels remained high during the vitellogenic stages, while GPα and LHβ mRNA levels peaked at the end of vitellogenesis. The expression of all three GtH subunits decreased during the post-spawning period. These results suggest that follicle-stimulating hormone (FSH) is involved in vitellogenesis, while luteinizing hormone (LH) functions during final oocyte maturation (FOM). Both GPα and FSHβ mRNA levels remained high during the FOM stages of the spawning cycle and increased further just after spawning. Thus, FSH synthesis may be strongly activated just after spawning to accelerate vitellogenesis in preparation for the next spawning. Alternatively, LHβ mRNA levels declined during hydration and then increased after ovulation. This study demonstrates that chub mackerel are a good model for investigating GtH functions in multiple spawning fish.