Selection of In Vitro-Matured Porcine Oocytes Based on Localization Patterns of Lipid Droplets to Evaluate Developmental Competence

Localization patterns of lipid droplets in the cytoplasm of porcine oocytes were evaluated as a novel marker for in vitro maturation (IVM) of oocytes with high developmental competence. Porcine oocytes were cultured in TCM-199, which is a complete synthetic medium, for 44 h at 38.5 C. Localization patterns were divided into 2 classes: lipid droplets localized uniformly in the whole cytoplasm (class I) and those that were centrally located (class II). After IVM in TCM-199, 60% of matured oocytes exhibited the class II pattern. To investigate the relation between the distribution of lipid droplets and the developmental rate of the oocyte, the developmental rates of class I and class II oocytes were compared after in vitro fertilization (IVF). Class II oocytes showed a significantly higher rate of blastocyst development than class I oocytes. These results suggest that porcine oocytes with high developmental competence can be selected based on the localization patterns of lipid droplets.     

Characteristic Variation of Concentration and Chemical Form in Sulfur, Nitrate, Ammonium, and Chloride Species Observed at Urban and Rural Sites of Japan

A field survey on the concentration of chemical species in particulate matter and gaseous compounds at two monitoring sites with different site classifications (urban and rural) was conducted over three years. Total (particulate matter + gaseous compounds) concentrations at the rural site were significantly lower than those at the urban site for all species (sulfur $\left( {{\text{SO}}^{{{\text{2 - }}}}_{{\text{4}}} {\left( {\text{p}} \right)}} \right.$ and SO₂(g)), nitrate ${\text{(NO}}_{{{\text{3}}^{{\text{ - }}} }} {\left( {\text{p}} \right)}$ and HNO₃(g)), ammonium ${\text{(NH}}_{{{\text{4}}^{{\text{ + }}} }} {\text{(p)}})$ and ammonia (NH₃(g)), and chloride (Cl^? (p) and HCl (g))), which is thought to reflect classification of the site. The difference in the sulfur concentration at the urban and rural sites was characterized by the difference in SO₂ (g) concentration. Further, a clear seasonality was observed for the nitrate species. The HNO₃ (g) concentration was high in the summer compared with other seasons at both the urban and rural sites. The ${\text{NH}}_4^ + \left( {\text{p}} \right)$ concentration levels were approximately the same as those of NH₃ (g) at both sites. The molar ratios of the particulate matter concentration to the total concentration showed different characteristics; the nitrate, ammonium and ammonia, and chloride species showed a clear seasonal variation: low in summer and high in winter and the values were similar regardless of the site. On the other hand, the sulfur species showed constant values at both the urban and rural sites, however the concentrations were significantly different for the two sites. Ammonium accounted for the largest proportion of cations in the particulate matter, regardless of the site classification. In contrast, ${\text{SO}}_4^{2 - } \left( {\text{p}} \right)$ accounted for the largest proportion of anions at the rural site, whereas ${\text{NO}}_3^ - \left( {\text{p}} \right)$ was comparable to ${\text{SO}}_4^{2 - } \left( {\text{p}} \right)$ at the urban site. Ammonia accounted for the largest proportion of all chemical species at both sites. Seasonal analysis of the proportional distribution in particulate matter and gaseous compounds provides information on atmospheric conditions.     

Agronomic evaluation and identification of potential cowpea (Vigna unguiculata L. Walp) genotypes in South Africa

Cultivar development of cowpea with increased grain yield depends on evaluation and selection of genetically unique and complementary breeding lines. The objective of this study was to evaluate cowpea collections using agronomic traits to select promising parents to develop breeding population. Twenty-two cowpea genotypes were field evaluated using a randomised complete block design with three replications in two locations in South Africa. Analysis of variance revealed significant genotypic (G) and locations (L) differences for all agronomic traits evaluated. Significant differences were also observed for most of the traits due to G X L, G X Year (Y), L X Y and G X L X Y interactions effects. The genotypes tested at two locations showed considerable genetic variation for the following agronomic traits that varied for number of leaves (NL) (20.08–179.78), leaf length (LL) (12.36–67.72 cm), leaf width (LW) (4.78–22.02 cm), number of branches (NB) (6.81–13.85), number of pods per plant (NPPP) (14.33–54.04), number of seeds per pod (NSPP) (11.7286.28), pod diameter (PD) (5.75–22.73 mm), plant height (PH) (6.57–233.71 cm), pod length (PL) (3.94–100.92 mm), hundred seed weight (HSWt) (8.11–49.52 g) and grain yield (GY) (0.10-1.48 t ha^?1). Grain yield was significant and positively correlated with all of the traits except leaf length. The principal component analysis (PCA) identified four principal components (PCs) contributing to 73.62% of the total explained variation amongst the tested genotypes. The selected cowpea genotypes for most of agronomic characteristics are useful and candidate genetic resources for the development of breeding population in cowpea breeding and enhanced production and productivity for food and nutrition in the country.     

An additional life-history tactic of masu salmon: Migration of parr to coastal habitats

Alternative life-history tactics of masu salmon Oncorhynchus masou are well documented. Subsequent to the freshwater parr stage (age ≥ 1 + years), an anadromous form migrates to the sea after smolting, while a resident form matures without seaward migration. In addition to this typical migratory dimorphism, anecdotal reports based on field observations have indicated that some underyearling masu salmon use estuarine waters. However, no empirical evidence indicates saltwater utilisation and subsequent survival in the early parr stage. Here, we used otolith microchemistry to examine whether a portion of masu salmon parr in northern Japan enters coastal habitats. The otolith Sr:Ca ratios of most juveniles collected from six rivers had consistently low values, indicating that masu salmon parr inhabiting these rivers stay only in freshwater. In contrast, in individuals from a steep-gradient river the Sr:Ca ratios increased at about a 200-µm distance from the otolith core. These results suggest that some masu salmon parr might use brackish water or sea water temporarily. In addition, three masu salmon parr were found in another steep river where a culvert located only ten metres from the river mouth completely blocked upstream migration for spawning. The Sr:Ca ratios in these fish increased at about >200 µm from the otolith core, indicating the parr had immigrated to the non-natal river from the sea. Such flexible behaviour at an early life stage may contribute to the spatial expansion of masu salmon, and the movement could moreover help to stabilise its population dynamics.     

Preimplantation development of embryos in labrador retrievers

Preimplantation development of canine embryos is not well understood. To understand the timing of preattachment embryogenesis relative to the luteinizing hormone (LH) surge, early embryonic development was examined in Labrador Retrievers after artificial insemination. The embryos migrated from the oviduct to the uterus beginning on day 11 after the LH surge. This transport must be completed within 24 h. By day 13 after the LH surge, all of the embryos had moved and were localized in the uterus. The embryos developed to the morula stage within 11-13 days and to the blastocyst stage within 14 days after the LH surge, respectively. These findings add to the current understanding concerning the physiology of preimplantation development and should help further develop assisted reproductive techniques in canine species, such as cryopreservation and subsequent embryo transfer.     

Downregulation of male-specific cytochrome P450 by profenofos

The health hazards of individual organophosphorus insecticides have been characterized by their acute toxicity, mainly by investigating their cholinesterase inhibition. However, the chronic effects of most of these toxicants on the drug-metabolizing enzymes have not been investigated. Profenofos (O-4-bromo-2-chlorophenyl O-ethyl S-propyl phosphorothioate) is an organophosphorus pesticide widely used in cotton cultivation. In the present study, we investigated the effect of profenofos on male-specific cytochrome P450 (CYP) enzymes in adult Wistar rats. We orally administered 17.8 mg/kg body weight, twice weekly for 65 days. Profenofos downregulated levels of hepatic and testicular CYP2C11 and CYP3A2 mRNA and protein expression. Testicular aromatase (CYP19A) mRNA was decreased in the profenofos-treated rats compared to controls. Overall, the present study suggests that profenofos acts as an endocrine disruptor of male-specific CYP enzymes and affects testosterone concentration, which implicates its deleterious effects on animal or human males chronically exposed to organophosphorus pesticide.     

Effects of resveratrol treatment on mitochondria and subsequent embryonic development of bovine blastocysts cryopreserved by slow freezing

This study evaluated the effects of cryopreservation by slow freezing on the mitochondrial function, DNA integrity, and developmental ability of bovine embryos and examined whether resveratrol treatment of the frozen‐thawed blastocysts improved embryonic viability. In vitro produced bovine embryos were subjected to slow freezing. After thawing, the ATP content and mitochondrial DNA integrity (mtDNA), determined by real‐time PCR targeting short and long mitochondrial sequences, was found to be lower in frozen‐thawed embryos than in fresh embryos, and mtDNA copy number was significantly reduced during the 24‐hr incubation post warming. Furthermore, immunostaining against double‐strand DNA revealed DNA damage in frozen‐thawed embryos. When frozen‐thawed embryos were incubated in the medium containing 0.5 µM resveratrol, SIRT1 expression, and survival rate of the embryos significantly improved compared with the vehicle‐treated embryos. In addition, cell‐free mtDNA content in medium was higher in case of resveratrol‐treated embryos than of vehicle‐treated embryos. In conclusion, slow freezing affects mitochondrial integrity and function in the blastocysts. In the frozen‐thawed embryos, mitochondria were removed during post‐thawing incubation and resveratrol enhanced the process, resulting in improved survivability of the embryos.     

Parent‐of‐origin effects on carcass traits in Japanese Black cattle

Variances caused by the differential expression of paternally and maternally imprinted genes controlling carcass traits in Japanese Black cattle were estimated in this study. Data on marbling score (BMS), carcass weight, rib thickness, rib‐eye area (REA) and subcutaneous fat thickness (SFT) were collected from a total of 13,115 feedlot steers and heifers in a commercial population. A sire–maternal grandsire model was used to analyse the data, and then, imprinting parameters were derived by replacing the genetic effect of the dam with the effect of the maternal grandsire in the imprinting model to calculate the genetic parameter estimates. The proportions of the total genetic variance attributable to imprinted genes ranged from 8.7% (SFT) to 35.2% (BMS). The remarkably large imprinting variance of BMS was mainly contributed by maternally expressed inheritance because the maternal contribution of the trait was much larger than that of the paternal trait. The parent‐of‐origin effect originating from maternal gene expression was also observed for REA. The results suggested the existence of genomic imprinting effects on the traits of the Japanese Black cattle. Hence, the parent‐of‐origin effect should be considered for the genetic evaluation of Japanese Black cattle in breeding programmes.