Induction of ovine trophoblast cell fusion by fematrin‐1 in vitro

Endogenous retroviruses present in the genomes take a specific role in placental formation in various vertebrates, including bovine and sheep. Fematrin‐1, which is the envelope (Env) protein of bovine endogenous retrovirus found in bovine placenta, is involved in the formation of fetomaternal hybrid cells in cattle placenta. This study was conducted to clarify whether fematrin‐1 possesses fusogenic activity in trophoblast cells. Another question is whether Env proteins only have species‐specific activity or not. For this, fematrin‐1 gene was transfected in ovine trophoblast cells, and we examined fusogenic activity with Cos‐7 cells. Although fematrin‐1 fusogenic activity was detected in both neutral and acidic pH conditions, acidic condition significantly enhanced it. These activities were rather weaker than those of vesicular stomatitis virus G protein as a positive control. However, the ratio of fematrin‐1 and vesicular stomatitis virus G protein fusion index was confirmed similar to those in the previous reports. Some fusion cells showed multinucleate cells. These results imply that fematrin‐1 is involved in the formation of trophoblast hybrid cells even in different species trophoblastic cells.     

Geographical differentiation and diallel analysis of seed dormancy in barley

Seed dormancy is one of the most important parameters affecting the malting process and pre-harvest sprouting in barley (Hordeum vulgare L.). Variation of seed dormancy in 4365 cultivated and 177 wild barley (ssp. spontaneum) accessions derived from different regions of the world was investigated in Okayama University, Kurashiki, Japan. Seed dormancy of each accession was estimated from their germination percentages at 0, 5, 10 and 15 weeks post-harvest after-ripening periods. All of the wild barley accessions showed less than 10% germination at 0 week after-ripening period. Level of seed dormancy in 4365 cultivated barley accessions showed a clear geographical differentiation. Seventy seven percent of Ethiopian accessions showed high germination percentages, while 86% of Japanese, Turkish and North African accessions showed low germination percentages at 0 week after-ripening period. A half diallel cross using eleven barley accessions with different level of dormancy revealed that seed dormancy was predominately controlled by additive gene effects. These results suggest that large genetic diversity for seed dormancy in barley is explained as different levels of additive accumulation of genetic factors. Barley varieties showing appropriate dormancy could be developed by crossing among barley germplasm accessions used in the present study.     

Early pregnancy diagnosis by means of ultrasonography as a method of improving reproductive efficiency in goats

Eighteen cyclic Shiba goats were used in this study. Estrus was synchronized with a single injection of 125 microg of a synthetic analogue of prostaglandin F(2)alpha (PGF(2)alpha) after detection of at least one corpus luteum by B-mode ultrasonography. Blood samples were collected from each animal on days 0, 7 and 21 post-mating for progesterone assay. Animals in estrus were either allowed to be mated by fertile bucks twice during estrus (group I; n=12) or not at all (group II; n=6). Ultrasonographic examinations were performed transrectally or transabdominally using a real-time B-mode scanner equipped with a 7.5 or 5 MHz transducer. All animals exhibited estrus 56.0 +/- 2.7 h after injection of PGF2alpha. The results show that the accuracy of the progesterone assay in diagnosing pregnancy on day 21 after mating was 80% for pregnancy and 100% for non-pregnancy, retrospectively. Ultrasonographic examinations showed that gestational sac and embryos heartbeats were detected on days 20.2 +/- 0.6 and 24.3 +/- 0.7 of gestation, respectively. Placentomes were detected on day 35.4 +/- 1.0 of gestation as small nodules (0.7 +/- 0.2 cm in size). At two months pregnancy, skeletal structures like skull, thorax and long bones were clear. Biparietal diameter of the skull and length of long bones could be used as an estimate of gestational age. The accuracy of detection of fetal number using real-time B-mode ultrasonography was 91.7% on day 60 of gestation. In conclusion, progesterone assay at day 21 post-mating (cut-off value, 1 ng/ml) can be used for pregnancy diagnosis in goats. However, B-mode transrectal ultrasonography was more efficient due to detection of embryo and confirmation of its viability by heartbeats. In addition, fetal number and gestational age could be determined only by ultrasonography.     

Effect of iron-quercetin complex on reduction of nitrite in in vitro and in vivo systems

This study investigated whether reducing agents such as quercetin and iron(II) facilitate formation of nitric oxide (NO) gas from orally ingested nitrite in an vivo study. When 3 mg/kg Na (15)NO2 was orally administered to rats with or without iron(II) or quercetin, Hb (15)NO, which is indicative of systemic (15)NO, was detected in the blood, with the maximum blood concentration of Hb (15)NO at 15 min after nitrite or nitrite plus quercetin treatment, whereas after administration of nitrite plus iron(II) or nitrite plus iron(II) and quercetin, the time was shortened to 10 min. Interestingly, iron(II), quercetin, or iron(II) plus quercetin did not affect the total amount of Hb (15)NO generated from orally administered Na (15)NO2. However, the systemic nitrite concentration was significantly decreased in the presence of iron(II) or iron(II) plus quercetin. These results may indicate that iron(II) is critical to the generation of NO gas from nitrite, whereas quercetin contributed little under the in vivo experimental conditions.     

Passive immunoneutralization of endogenous inhibin increases ovulation rate in miniature Shiba goats

We reported previously that passive immunization against inhibin enhances follicular growth and increases the ovulation rate. However, the ovulation rate was not comparable to the number of follicles. Therefore, the aim of this study was to attempt to increase the ovulation rate by increasing the interval between inhibin immunization and PGF2alpha injection. Five miniature Shiba goats were treated with 10 ml inhibin antiserum (inhibin-AS) developed against [Tyro30]-inhibin alpha (1-30). A control group (n=5) was treated with normal goat serum. All animals were injected intramuscularly with 125 microg PGF2alpha 72 h after treatment to induce estrus and ovulation. Blood samples were collected for hormonal assay and the ovulation rate was determined by laparotomy. In contrast to the control group, there was a significant increase in plasma concentrations of FSH in the immunized group. After luteolysis, plasma concentrations of estradiol-17beta increased markedly to a preovulatory peak about 2 folds higher (P<0.01) than that of controls. In addition, the ovulation rate was greater in the immunized group (14.4 +/- 2.2) than in the control group (2.2 +/- 0.6), and the mean number of follicles > or = 4 mm in diameter was 10.0 +/- 0.8 in the inhibin-AS group compared with 2.4 +/- 0.3 in control group. The present results demonstrate that immunoneutralization of endogenous inhibin increased FSH secretions in miniature shiba goats. The increased FSH secretion enhanced follicular growth and increased the ovulation rate. Additionally, increasing the interval between inhibin-AS and PGF2alpha injections (to 72 h) resulted in a greater ovulation rate compared with the previous protocol (48 h). Therefore, inhibin-AS treatment proved to be an effective alternative to exogenous gonadotropin methods for induction of superovulation in goats.     

Migration and spawning behavior of the greater amberjack Seriola dumerili in eastern Taiwan

Greater amberjack (Seriola dumerili) is an important fishery resource with a circumglobal distribution from tropical to temperate waters. Here, we investigated the spawning migration and habitat utilization of S. dumerili in the East China Sea (ECS). Archival tags were attached to 22 adult fish to examine their horizontal and vertical movements and estimate the spawning ground. S. dumerili were captured and released in the coastal waters of eastern Taiwan on November of 2016 and 2017. Information from seven pop-up satellite archival tags and seven depth–temperature recorders was recovered. Almost all of the fish stayed in the Taiwanese exclusive economic zone. Most individuals moved from released site to the southern edge of the ECS and showed behavior associated with the topographic features in winter (November to December). These phenomena may be related to foraging and be driven by oceanographic features such as the seasonal monsoon and the Kuroshio. The fish then migrated to the south offshore area of Taiwan in January and February. During their southward migration, the fish experienced a slowly elevated water temperature regime (SETR), which is one of the environmental factors that induce final oocyte maturation. In the spawning season (February to April), tagged females exhibited continuous diel vertical movements (DVMs) after experiencing the SETR. These continuous DVMs were observed over a wide geographic range from north to south in the Kuroshio off eastern Taiwan. Our study demonstrated that the putative spawning ground of S. dumerili must extend further in a north–south direction than predicted in a previous study.     

Use of acceleration loggers in aquaculture to determine net-cage use and field metabolic rates in red sea bream <Emphasis Type="Italic">Pagrus major</Emphasis>

We investigated the usefulness of acceleration loggers in aquaculture by examining net-cage use and metabolic rates in red sea bream, Pagrus major. First, the fish’s metabolic rate (mg O₂ kg⁻¹ min⁻¹) was measured with the logger in a swim tunnel at designated water velocities. We found that metabolic rate could be expressed by using a linear regression model of the activity rate index (unitless min⁻¹) derived from acceleration data. Using this equation, the field metabolic rates of three fish in a net cage were monitored and were estimated at 14.1–15.0 kcal kg⁻¹ day⁻¹. The results suggested that 15–19% of energy from satiation feeding ration was consumed for metabolism and activity in the net cage. The loggers showed orderly net-cage use by the fish. Tagged individuals used the whole cage from surface to bottom, but individual fish that preferred the surface area rarely used the bottom, and vice versa. Metabolic rate increased significantly with distance of the fish from their preferred depths. The logger provided information on the physiological and behavioral responses of fish in a given breeding system, and its use should contribute to the design of practical aquaculture systems.     

Effects of hypothyroidism on gonadal function after transition of short day photoperiod in male golden hamsters (Mesocricetus auratus)

Thyroid hormones permit the annual reproductive transition of seasonal breeders. Although, precise function of thyroid hormones in seasonal breeding is not well understood. In the present study, we examined effects of hypothyroidism on the hypothalamus-pituitary-gonadal axis in adult male golden hamsters after transition of the short-day photoperiod (SD; 8 h light: 16 h dark) condition. We confirmed that hypothyroid, which had been induced by administration of thiouracil in drinking water for 4 weeks, did not have direct effects on testes in male hamsters under the long-day photoperiod. Plasma concentrations of free T3 and T4 decreased 15 weeks after transition of SD condition. Plasma concentrations of testosterone in the hypothyroid group decreased earlier than in the control group after the transition from LD to SD. In animals treated with testosterone after castration, plasma concentrations of LH in the hypothyroid group decreased earlier than in the control group after the transition of SD. On the other hand, pituitary response to GnRH for LH release did not change in castrated hamsters as a result of hypothyroidism. These results suggest that thyroid hormones act the hypothalamus and might be required to maintain GnRH secretion in male golden hamsters.     

Low expression of the antibacterial factor L‐amino acid oxidase in bovine mammary gland

In the mouse, L‐amino acid oxidase (LAO) produces hydrogen peroxide by utilizing free amino acids and is a proven antibacterial factor in mammary glands. Mastitis, a bacterial infection of the mammary gland, is the most frequent disease in dairy cattle. Here, we investigate whether LAO is expressed in the mammary gland of dairy cattle and is antibacterial. In dairy cattle, the expression level of LAO mRNA in the mammary gland was considerably lower than that in mice, and LAO activity was not observed in cattle milk that produced hydrogen peroxide. The expression of LAO mRNA was also low in Japanese Black cattle, the same as in Holstein cattle. A higher LAO mRNA expression was observed in the mastitis glands than in the lactating glands. Furthermore, spleen and lymph nodes expressed high levels of LAO mRNA in dairy cattle. We conclude that mammary glands in dairy cattle have lower ability to express the LAO gene compared to that in mice, which may result in a high incidence of mastitis.     

Role of polyamines in peach fruit development and storage

To investigate the role of polyamines in pre- and post-harvest fruit development of 'Akatsuki' peach (Prunus persica (L.) Batsch.) we measured polyamine concentrations, activities of polyamine biosynthetic enzymes and expression of genes encoding these enzymes. Concentrations of the free polyamines, putrescine (Put), spermidine (Spd) and spermine (Spm) in pre-harvest fruit peaked 16 days after full bloom (DAF) and then progressively decreased until harvest with the exception of Put, which showed a second peak at 94 DAF, just before the onset of ethylene production. In post-harvest fruit, minor changes in concentrations of Spd and Spm were observed, whereas Put concentration peaked on the harvest day, followed by an abrupt decrease and a subsequent 2-fold increase, which was opposite to the fluctuating pattern of ethylene production. Activities of arginine decarboxylase (ADC) and ornithine decarboxylase (ODC) peaked during the first stage of fruit development and then decreased until 80 DAF, after which the activities were below detection limits, suggesting that Put is synthesized during the early stage of fruit development. Activity of S-adenosylmethionine decarboxylase (SAMDC) decreased progressively until the end of S2. Expression levels of five putative polyamine biosynthetic genes, ADC, ODC, SAMDC, spermidine synthase (SPDS) and spermine synthase (SPMS), in pre-harvest and post-harvest fruit did not coincide precisely with the observed changes in enzymatic activities and polyamine concentrations. The possible role of polyamines during peach fruit development and the relationship between polyamines and ethylene biosynthesis are discussed.