Reproductive biology of male skipjack tuna <Emphasis Type="Italic">Katsuwonus pelamis</Emphasis> (Linnaeus) in the tropical western and central Pacific Ocean

The reproductive biology of male skipjack tuna Katsuwonus pelamis was examined from May 2005 to December 2007 in the tropical western and central Pacific Ocean. Testis maturity was classified into five stages (i.e., immature, early maturation, mid maturation, late maturation, and regression) on the basis of morphological changes in the germinal epithelium. The testis duct system (TS), which was characterized by lobules that lack spermatocysts, store only sperm, and form an anastomosing network, was located under the main sperm duct. The TS was observed at all stages of maturity except the immature stage, and first appeared in individuals with a fork length (FL) of 38–40 cm. The estimated minimum size at first maturity was 35.5 cm FL and the size at 0.5 maturity was 40.7 cm in FL. It was difficult to clearly differentiate the testis maturity stage based solely on the gonad index (GI) class because each GI class contained mature fish. Monthly changes in testis height index and GI did not follow a seasonal cycle. Mature individuals dominated throughout the year. These results imply that male skipjack tuna in this area showed evidence of spawning activity throughout the year without a clear seasonal pattern.     

Motility and fertility of cryopreserved spermatozoa of the Japanese sea cucumber Apostichopus japonicus

We developed both a cryopreservation method for Japanese sea cucumber spermatozoa and an artificial fertilization method using post‐thaw spermatozoa. Twenty per cent dimethyl sulfoxide (DMSO), 16% foetal bovine serum, and 64% artificial seawater were suitable cryodiluent, and the diluent was pre‐cooled to 0°C. Semen was diluted with the solution and enclosed in a 250 μl straw, cooled to ?50°C at 10.4 ± 0.4°C/min, and immediately immersed in liquid nitrogen. Although this method showed the highest post‐thaw motility in all the conditions we examined, its post‐thaw motility was still less than approximately 15%. Artificial fertilization was carried out by adding post‐thaw semen with a cryodiluent to the oocytes. The fertilization rate of 200 oocytes/ml seawater increased with the amount of post‐thaw semen from 1 to 5 μl but showed a significant decrease at 25 μl. This decrease was considered to be due to DMSO in the cryodiluent, because the fertilization rate of the fresh semen decreased sharply when the DMSO concentration around the oocytes was 1.0% or more. Further improvement in increasing post‐thaw motility and lowering the cryoprotectant concentration is necessary for commercial‐scale artificial fertilization.     

Evaluation of changes in left ventricular myocardial function observed in canine myocardial dysfunction model using a two-dimensional tissue tracking technique

This study was conducted to assess the ability of two-dimensional tissue tracking (2DTT) to evaluate changes in left ventricular (LV) myocardial function associated with sustained high electrical pacing. Pacemakers were implanted at the right ventricular (RV) apex of five female Beagles, and sustained high electrical pacing of 250 beats per minute (bpm) was performed for three consecutive weeks. Conventional echocardiography and 2DTT were performed at baseline, and at every week for three weeks with pacing. The baseline parameters were then compared to those of weeks 1, 2, and 3. Three weeks of pacing resulted in significant reduction of radial and circumferential global strains (p < 0.001). Regional analysis revealed reduction of segmental strains in both radial and circumferential directions, as well as increased dyssynchrony after three weeks of pacing in the radial direction (p = 0.0007). The results of this study revealed the ability of 2DTT to measure radial and circumferential strains in dogs with sustained high-electrical pacing, and allowed assessment of global and regional myocardial function and the degree of dyssynchrony.     

Laminin binds to myostatin and attenuates its signaling

Myostatin is a growth and differentiation factor and acts as a negative regulator of skeletal muscle mass. Although the mechanism whereby myostatin controls muscle cell growth is mostly clarified, the regulation of myostatin activity after its secretion into the extracellular matrix (ECM) is still unclear. In the present study, we investigated the interaction between laminin and myostatin and the effect of laminin on myostatin signaling in vitro. The surface plasmon resonance assay showed that laminin bound to mature myostatin and activin receptor type IIB (ActRIIB), but did not bind to latency‐associated protein, which remains non‐covalently linked to mature myostatin. Furthermore, kinetic analysis demonstrated that the affinity of mature myostatin for laminin was similar to that for ActRIIB. Next, we examined the action of laminin on the myostatin signaling pathway using a conventional reporter assay. The luciferase activity of myostatin‐treated cells was repressed significantly (P < 0.05) by coincubation of laminin. These results suggest that laminin has a potential to regulate myostatin activity through binding to mature myostatin and/or its receptor ActRIIB.     

Study on structure and function of an earthen bund irrigation system in Northeast Thailand

In Northeast Thailand, which overlaps with an erosional plain that is not suitable for irrigation because of poor water resources and terrains that prevent efficient water distribution, farmers have long supplied water to surrounding paddy fields by blocking rivers completely with earthen bunds. Although such a traditional irrigation system fits well with the characteristics of Northeast Thailand, those who are concerned with modern irrigation development projects in Northeast Thailand seem to have been paying little attention to it. The present study was performed to facilitate development of more appropriate irrigation systems in Northeast Thailand by providing information regarding traditional irrigation methods. We investigated the traditional irrigation methods with earthen bunds at three study sites, and our results indicated that styles of irrigation vary with riverbed slope. The traditional irrigation system that is used in most of Northeast Thailand is different from ordinal weir irrigation, in that paddy fields are not irrigated by gravity flow but by backwater of earthen bunds. Our results suggest that the development of more successful irrigation systems would be possible through improvement of the design ideas of traditional irrigation methods.

Effects of rice feeding and carnitine addition on growth performance and mRNA expression of protein metabolism‐related genes in broiler grower chicks

This study was carried out to evaluate the nutritional effects of rice feeding and carnitine addition to a diet for broiler chicks. Thirty‐six male 10‐day‐old broiler chicks were assigned to one of the following four treatment groups: corn‐based diet (corn group), rice‐based diet (rice group), and each diet with added carnitine (100 ppm). The experimental period was 2 weeks. Rice feeding resulted in significantly higher growth performance (body weight gain and feed efficiency) compared to corn feeding. Carnitine addition also resulted in higher growth performance. Breast muscle and thigh muscle weight (g) were significantly higher in broiler chicks fed rice and those fed diets with added carnitine. Liver mRNA expression of IGF‐I was significantly higher in broiler chicks fed rice compared to those fed corn. There was no significant difference in mRNA expression of muscle atrogin‐1 or liver CPT‐I between broiler chicks fed rice and those fed corn, not between broilers chicks fed diets containing carnitine or not. Overall, these results show that rice feeding and carnitine addition improve the growth performance of broiler chicks by increasing mRNA expression of liver IGF‐I. In addition, carnitine action is not affected by different cereals (corn and rice).     

Genomic characterization and distribution of bovine foamy virus in Japan

Bovine foamy virus (BFV) is distributed through worldwide cattle herds. Although the biological features of BFV are not well understood, appearance of clinical manifestation by superinfection with other microorganisms is inferred. In Japan, reports of genomic characterizations and epidemiology of this virus are limited. In this study, we performed whole genomic sequencing of BFV strains Ibaraki and No.43, which were isolated in this country. Additionally, we investigated BFV in geographically distant four daily farms in Japan, to estimate the distribution of BFV and its correlation to bovine leukemia virus (BLV). BFV was distributed throughout Japan; the average positive rate was 12.7%. The nucleotide sequence identities of the isolates were 99.6% when compared with BFV strain isolated in the USA. The phylogenetic tree using env gene sequence showed strains Ibaraki, No.43 and Kagoshima were sorted in the same cluster including the USA and Chinese strains, while Hokkaido strain was in the other cluster including European strains. Although no clear correlation between BFV and BLV could be found, BFV and BLV infections were likely to increase with ages. Our data on epidemiology and characteristics of BFV will provide important information to reveal biological features of BFV.     

Changes in nitrogen transformation in forest soil representing the climate gradient of the Japanese archipelago

Net nitrogen transformation was investigated under different climate conditions by soil transplantation and in situ incubation of forest surface soils using the resin-core method. Selected conditions were considered to reflect those of the natural climate gradient in the Japanese archipelago. Study sites were established in natural forests in northern Hokkaido (Uryu), northern Kanto (Kusaki), central Kinki (Kamigamo), and southern Kyushu (Takakuma), representing the northernmost to the southernmost island regions of Japan. Field experiments comparing soils incubated at “native” and “transplanted” sites were conducted from June 2008 to May 2009. Net production, accumulation, and leaching of soil ammonium (NH₄ ⁺) and nitrate (NO₃ ⁻) were measured at each of the sites during the growing season (June–October), the dormant season (November–May), and throughout the year. Net nitrate production was highest in Kusaki soil, especially during the growing season, whereas net ammonium production was highest in Uryu soil, the coldest site, especially during the dormant season. Net nitrate production increased significantly in soils transplanted to a warmer climate during the growing season. However, net ammonium production increased in soils transplanted to colder climates during the dormant season. These findings indicate that, with the exception of the infertile soil samples from Kamigamo, the range of natural climates in Japan has a significant effect on nitrogen availability in surface soil. In addition, the original characteristics of the nitrogen cycle of the surface soil from each native site were retained, even when marked changes in soil temperature (approximately 8°C) occurred after transplantation.     

Developmental ability of vitrified mouse oocytes expressing water channels

Previously, we showed that the exogenous expression of aquaporin 3 (AQP3), an aquaglyceroporin, improved the tolerance of mouse oocytes to vitrification with a glycerol-based solution. In the present study, we examined conditions suitable for the expression of AQP3 and the ability of vitrified oocytes to develop in vitro and in vivo after fertilization. After only partial remove of cumulus cells, immature mouse oocytes (germinal vesicle stage) were injected with 5, 10 or 20 pg of AQP3 cRNA and cultured for 12 h for maturation. When matured oocytes were vitrified with a glycerol-based solution, 57-61% survived regardless of the amount of cRNA injected (5-20 pg). By contrast, no oocytes injected with water (control) survived. When the zona pellucida was removed from the vitrified oocytes and the oocytes were then fertilized in vitro and cultured, the proportions that were fertilized and developed into blastocysts were higher when the amount of cRNA injected was 5 pg than 10-20 pg. When 16 blastocysts were transferred to a pseudopregnant mouse, 5 developed to term, demonstrating that oocytes vitrified after injection of AQP3 cRNA retained the ability to develop to term. The water-permeability of cRNA-injected oocytes was higher than that of control oocytes from the maturing stage to the 1-cell zygote stage, whereas glycerol-permeability was higher only at metaphase II. This indicates that AQP3 was expressed for a relatively short period of time. These results suggest that the transient expression of water/cryoprotectant channels is effective for cryopreserving cells that have low membrane-permeability, such as mammalian oocytes.