Dynorphin-Kappa Opioid Receptor Signaling Partly Mediates Estrogen Negative Feedback Effect on LH Pulses in Female Rats

Accumulating evidence suggests that the arcuate nucleus (ARC) kisspeptin/neurokinin B (NKB)/dynorphin (KNDy) neurons play a role in estrogen negative feedback action on pulsatile gonadotropin-releasing hormone (GnRH)/luteinizing hormone (LH) release. The present study aimed to determine if dynorphin (Dyn) is involved in estrogen negative feedback on pulsatile GnRH/LH release. The effect of the injection of nor-binaltorphimine (nor-BNI), a kappa-opioid receptor (KOR) antagonist, into the third cerebroventricle (3V) on LH pulses was determined in ovariectomized (OVX) adult female rats with/without replacement of negative feedback levels of estradiol (low E₂). The mean LH concentrations and baseline levels of LH secretion in nor-BNI-injected, low E₂-treated rats were significantly higher compared with vehicle-treated controls. On the other hand, the nor-BNI treatment failed to affect any LH pulse parameters in OVX rats without low E₂ treatment. These results suggest that Dyn is involved in the estrogen negative feedback regulation of pulsatile GnRH/LH release. The low E₂ treatment had no significant effect on the numbers of ARC Pdyn (Dyn gene)-,Kiss1- and Tac2 (NKB gene)-expressing cells. The treatment also did not affect mRNA levels of Pdyn and Oprk1 (KOR gene) in the ARC-median eminence region, but significantly increased the ARC kisspeptin immunoreactivity. These findings suggest that the negative feedback level of estrogen suppresses kisspeptin release from the ARC KNDy neurons through an unknown mechanism without affecting the Dyn and KOR expressions in the ARC. Taken together, the present result suggests that Dyn-KOR signaling is a part of estrogen negative feedback action on GnRH/LH pulses by reducing the kisspeptin release in female rats.     

Motility and fertility of boar semen after liquid preservation at 5°C for more than 2 weeks

This study investigated the effects of skim milk on the quality and fertility of boar spermatozoa under long‐term chilled preservation. Semen samples were stored in Modena solution supplemented with 0 (control) to 50 mg/mL skim milk at 5°C for 4 weeks; spermatozoa stored with 7.5 and 15 mg/mL of skim milk (7.5‐SM and 15‐SM groups, respectively) exhibited significantly higher motility indices than those of the control group up to 3 weeks (P < 0.05), and the 7.5‐SM group showed improved motility indices even after 4 weeks (P < 0.05). In vitro fertilization using spermatozoa in the 7.5‐SM and 15‐SM groups stored at 5°C for 2 weeks showed significantly higher fertilization rates of spermatozoa and the development rates to blastocyst than the control group (P < 0.05), and the 7.5‐SM group showed similar rates of fertilization and blastocyst formation in the fresh non‐stored spermatozoa group. After artificial insemination using spermatozoa stored for 2 weeks in the 7.5‐SM group, healthy piglets were obtained. Boar spermatozoa can be stored at 5°C in a Modena solution containing skim milk. Supplementation of 7.5 mg/mL skim milk improves boar spermatozoa motility and fertility even after liquid preservation at 5°C for 2 weeks.     

Characterization of a novel potyvirus tentatively named <Emphasis Type="Italic">Ornithogalum</Emphasis> virus 3, successfully isolated from <Emphasis Type="Italic">O. thyrsoides</Emphasis> co-infected with two other potyviruses by single-aphid inoculation

A potyvirus tentatively named Ornithogalum virus 3 (OV-3) was successfully isolated by single-aphid transmissions from O. thyrsoides mix-infected with OV-3, Ornithogalum mosaic virus (OrMV) and Ornithogalum stripe mosaic virus (OrSMV). OV-3, a flexuous, rod-shaped particle of ca. 690 nm, was sap and aphid transmissible. The virus had a narrow host range and caused necrotic mosaic on O. thyrsoides under cold conditions. We therefore propose the name Ornithogalum necrotic mosaic virus (OrNMV) for OV-3. A synergistic increase in symptom severity was apparent on O. thyrsoides mix-infected with OrSMV/OrNMV, but not with either OrMV/OrNMV or OrMV/OrSMV. The nucleotide sequence data reported is available in the DDBJ/EMBL/GenBank databases under accession number AB282754.     

Basin-scale distribution pattern and biomass estimation of Japanese anchovy Engraulis japonicus in the western North Pacific

The distribution pattern and biomass of the Japanese anchovy Engraulis japonicus in the offshore region of the western North Pacific (north of 35°N and west of 170°E) were studied using a quantitative echosounder. This is the first attempt at such a study in this region. Data were collected in summer from 2004 to 2007. The biomass was estimated using data collected at 38?kHz. Species compositions in the backscatterings from pelagic fish were assigned based on the results of trawl hauls taking account of sea surface temperature (SST). Japanese anchovy tended to be high density to the west of 153°E and were distributed in an SST range of 9?C24?°C. Although the temporal and spatial coverage of the survey differed each year, at least 1.5?C3.4 million tons of Japanese anchovy were present in the survey area between 2004 and 2007. To take account of the spatial coverage of the survey each year, the most reliable biomass estimate for this region in the time period was 3.4 million tons (coefficient of variation?0.22).     

Oceanographic factors affecting interannual recruitment variability of Pacific saury (Cololabis saira) in the central and western North Pacific

Pacific saury (Cololabis saira) has a short life span of 2 years and tends to exhibit marked population fluctuations. To examine the importance of sea surface temperature (SST) and mixed layer depth (MLD) as oceanographic factors for interannual variability of saury recruitment in early life history, we analyzed the relationship between abundance index (survey CPUE (catch per unit of effort)) of age‐1 fish and the oceanographic factors in the spawning and nursery grounds of the previous year when they were born, for the period of 1979–2006, in the central and western North Pacific. Applying the mixture of two linear regression models, the variability in the survey CPUE was positively correlated with previous year's winter SST in the Kuroshio Recirculation region (KR) throughout the survey period except 1994–2002. In contrast, the survey CPUE was positively correlated with the previous year's spring MLD (a proxy of spring chlorophyll a (Chl‐a) concentration) in the Kuroshio‐Oyashio Transition and Kuroshio Extension (TKE) during 1994–2002. This period is characterized by unusually deep spring MLD during 1994–1997 and anomalous climate conditions during 1998–2002. We suggest that saury recruitment variability was generally driven by the winter SST in the KR (winter spawning/nursery ground), or by the spring Chl‐a concentration (a proxy of prey for saury larvae) in the TKE (spring spawning/nursery ground). These oceanographic factors could be potentially useful to predict abundance trends of age‐1 saury in the future if the conditions leading to the switch between SST and MLD as the key input variable are elucidated further.     

Comparison of the nutritional transition date distributions of marbled sole larvae and juveniles in Hakodate Bay, Hokkaido

To examine survival processes in marbled sole Pseudopleuronectes yokohamae larvae, nutritional transition date distributions (NTDDs) were compared between larvae and large juveniles (LJs) in Hakodate Bay from 2001 to 2003. NTDDs were used instead of hatch date distributions because the onset of increment formation coincides with the timing of the transition from endogenous to exogenous nutritional sources for marbled sole. LJs were defined as being large enough to be safe from predation by the sand shrimp Crangon uritai. In 2001 and 2002, the ranges of the NTDDs of LJs largely overlapped with those of the larvae. However, in 2003, the early cohort (those whose otolith rings formed before 17 March) disappeared from the NTDDs during and after April, and the NTDDs of LJs were heavily biased toward a later period compared to those of the larvae. In 2003, Oyashio coastal water (<3°C and ≤33.0 salinity) unexpectedly intruded into the bay in late March and lowered the water temperature from 6.3°C on 6 March to 4.1°C on 25 March. One possible cause of mortality in the early cohort of 2003 may be feeding failure caused by the extremely low water temperature.     

Direct estimation of stock abundance of Pacific saury <Emphasis Type="Italic">Cololabis saira</Emphasis> in the northwestern Pacific Ocean using a mid-water trawl

Drifting gill-nets were for a long time the main research gear for investigating the distribution of Pacific saury Cololabis saira. However, it has proven to be difficult to estimate the absolute abundance of saury using drifting gill-nets due to a lack of knowledge on the survey areas swept by these gill-nets. A mid-water trawl stock abundance estimation has the advantage of being able to estimate the absolute stock abundance before the commercial fishing season begins, whereas the conventional stock abundance estimation based on catch per unit efforts of commercial fisheries must be done after the commercial fishing season has ended. To confirm the practicality of a mid-water trawl to estimate the stock abundance of saury, we conducted a comparative survey in the northwestern Pacific Ocean using both a mid-water trawl and drifting gill-nets. We found that the geographic distribution pattern of catch per unit effort for both the mid-water trawl and driftnets was similar. Using the area-swept method and data on the fishing efficiency of the mid-water trawl obtained in a previous study, we were able to estimate the stock abundance plus confidence limits within the research areas. Our results suggest that the size composition of saury sampled by the mid-water trawl can be considered to approximate the actual size composition of saury.     

Immunohistochemical analysis of the distribution of classical swine fever (CSF) viral antigen in boar-pig hybrids and pigs four weeks after infection

We detected the classical swine fever virus (CSFV) antigen in three boar-pig hybrids (hybrids) and three pigs. All animals were experimentally infected with CSFV strain JPN/27/2019 to optimize diagnostic sampling and risk assessment of virus dissemination. Two hybrids died 17- and 19-days post-inoculation (dpi). The other animals were euthanized at 28 dpi. The detection of CSFV antigen at 28 dpi in epithelial cells of the apocrine sweat and sebaceous glands in the skin, salivary glands, mucosal epithelial cells in the rectum, and epithelial cells in the kidney and urinary bladder, suggests that CSFV persists in these tissues and spreads via sweat, saliva, feces, and urine for at least 4 weeks. These findings reveal that hybrids and pigs represent a high risk of virus dissemination four weeks after infection with CSFV strain JPN/27/2019. Prominent CSFV antigens were also detected in hair follicles of the skin. These results suggest that postmortem sampling of animal skin may be effective for CSF diagnosis and can be used to develop a rapid and easy diagnostic method using hair follicles.     

Effect of maturation culture period of oocytes on the sex ratio of in vitro fertilized bovine embryos

It has been suggested that the maturational stage of oocytes at time of insemination influences the sex ratio of resulting embryos. However, there are very few reports concerning the relationship between the maturation culture period of oocytes and the sex ratio of resulting embryos. The objective of this study was to investigate the effects of in vitro maturation culture period for bovine oocytes on the sex ratio of in vitro produced blastocysts using a novel technique of loop-mediated isothermal amplification (LAMP). Cumulus-oocyte complexes were collected from the ovaries of slaughtered cows, and then matured in vitro for various periods (16, 22, 28, and 34 h). After maturation culture for each period, the oocytes were inseminated with frozen-thawed spermatozoa, and then cultured in vitro. Blastocysts were harvested on Day 7 after insemination, and the sex of the embryos was examined using the LAMP method. The rates of oocytes matured to the metaphase II stage were significantly lower (P < 0.05) in the 16-h maturation group than in the other groups. The proportion of blastocyst formation after insemination was significantly higher (P < 0.05) in the 22-h maturation group than in the other groups. The proportion of male blastocysts increased with the increase in maturation culture period. The proportion of male blastocysts derived from oocytes matured for 34 h was significantly higher (P < 0.05) than from oocytes matured for 16 and 22 h. These results indicate that the sex ratio of in vitro fertilized embryos is apparently influenced by the maturation culture period of the oocytes.