Annual changes in testicular size and serum and fecal testosterone concentrations in male bharals, Pseudois nayaur

The serum and fecal testosterone (T) concentrations and testicular sizes of two male bharals (Pseudois nayaur) were determined for approximately one year. The profiles of the fecal T concentrations showed a similar tendency as the profiles of serum T concentrations, and there was a significant correlation between serum and fecal T concentrations (r=0.72). T concentrations rose drastically in October and decreased gradually until January. The maximum testicular size was observed between November and January. Semen collected between December and January was excellent in quality and comparable to domestic sheep and goats. The active periods of the testes were synchronized with the early breeding season of females.     

Mortality processes of hatchery‐reared Pacific bluefin tuna Thunnus orientalis (Temminck et Schlegel) larvae in relation to their piscivory

In mass culture of Pacific bluefin tuna Thunnus orientalis, yolk‐sac larvae of other species are fed as a major prey item to tuna larvae from 7 to 8 mm in total length. Marked growth variations in tuna larvae are frequently observed after feeding of yolk‐sac larvae, and this variation in the growth of tuna larvae is subsequently a factor leading to the prevalence of cannibalistic attacks. To elucidate details of the mortality process of hatchery‐reared tuna larvae after the initiation of yolk‐sac larvae feeding, we compared the nutritional and growth histories of the surviving (live) tuna larvae to those of the dead fish, found dead on the bottom of the tank, as direct evidence of their mortality processes. Cause of mortality of tuna larvae 3 and 5 days after the initiation of feeding of yolk‐sac larvae was assessed from nitrogen stable isotope and otolith microstructure analyses. Stable isotope analysis revealed that the live fish rapidly utilized prey fish larvae, but the dead fish had depended more on rotifers relative to the live fish 3 and 5 days after the initiation of feeding of yolk‐sac larvae. The growth histories based on otolith increments were compared between the live and dead tuna larvae and indicated that the live fish showed significantly faster growth histories than dead fish. Our results suggest that fast‐growing larvae at the onset of piscivory could survive in the mass culture tank of Pacific bluefin tuna and were characterized by growth‐selective mortality.     

Photosynthetic traits around budbreak in pre-existing needles of Sakhalin spruce (Picea glehnii) seedlings grown under elevated CO2 concentration assessed by chlorophyll fluorescence measurements

To assess the effects of elevated CO(2) concentration ([CO(2)]) on the photosynthetic properties around spring budbreak, we monitored the total leaf sugar and starch content, and chlorophyll fluorescence in 1-year-old needles of Sakhalin spruce (Picea glehnii Masters) seedlings in relation to the timing of budbreak, grown in a phytotron under natural daylight at two [CO(2)] levels (ambient: 360?μmol mol(-1) and elevated: 720?μmol mol(-1)). Budbreak was accelerated by elevated [CO(2)] accompanied with earlier temporal declines in the quantum yield of PSII electron transport (Φ(PSII)) and photochemical quenching (q(L)). Plants grown under elevated [CO(2)] showed pre-budbreak leaf starch content twice as high with no significant difference in Φ(PSII) from ambient-CO(2)-grown plants when compared at the same measurement [CO(2)], i.e., 360 or 720?μmol mol(-1), suggesting that the enhanced pre-budbreak leaf starch accumulation might not cause down-regulation of photosynthesis in pre-existing needles under elevated [CO(2)]. Conversely, lower excitation pressure adjusted for the efficiency of PSII photochemistry ((1?-?q(P)) F(v)'/F(m)') was observed in plants grown under elevated [CO(2)] around budbreak when compared at their growth [CO(2)] (i.e., comparing (1?-?q(P)) F(v)'/F(m)' measured at 720?μmol mol(-1) in elevated-CO(2)-grown plants with that at 360?μmol mol(-1) in ambient-CO(2)-grown plants), which suggests lower rate of photoinactivation of PSII in the elevated-CO(2)-grown plants around spring budbreak. The degree of photoinhibition, as indicated by the overnight-dark-adapted F(v)/F(m), however, showed no difference between CO(2) treatments, thereby suggesting that photoprotection during the daytime or the repair of PSII at night was sufficient to alleviate differences in the rate of photoinactivation.     

Inoculation with N2-generating denitrifier strains mitigates N2O emission from agricultural soil fertilized with poultry manure

Pelleted poultry manure is recommended for use with agricultural soil as a replacement for chemical fertilizers; however, application of the manure stimulates nitrous oxide (N₂O) emission from the soil through denitrification. To mitigate the N₂O emission caused by application of pelleted poultry manure, soil microcosms were set up; each microcosm was inoculated with one of the following N₂-generating denitrifier strains previously been isolated from paddy soil: Azoarcus, Dyella, Dechloromonas, Niastella, and Burkholderia. When pelleted poultry manure was incubated on its own, N₂O was produced by denitrification. In contrast, N₂O emission was significantly lowered when the manure was inoculated with most of the N₂-generating strains. In soil microcosms, N₂O was emitted during incubation after application of the pelleted manure, while N₂O flux was significantly lowered when the soil was inoculated with Azoarcus sp. KS11B, Niastella sp. KS31B, or Burkholderia sp. TSO47-3 on the 12th day of incubation. In addition, when pelleted manure was inoculated with the strains prior to application in the soil microcosms, the level of N₂O emission was significantly lowered to ca. 40–60 % that from the non-inoculated control. Our study provides the prototype of a technique that uses microbial technology to mitigate N₂O emission from agricultural soil fertilized with pelleted poultry manure.     

A basic experiment of coral culture using sexual reproduction in the open sea

Coral larvae, produced from a mass spawning event, were successfully settled on special stone settlement sticks and raised in situ for eventual transport to other reefs. The test area, Sekisei lagoon, Okinawa Prefecture, Japan, is located close to the warm Kuroshio current and is surmised to be the source from which major Japanese corals are derived. A total of 131 settlement sticks, with small holes in their sides to increase protection from grazing (4 mm in diameter and approximately 5 mm deep), were deployed in the lagoon the day before the coral’s mass spawning. After 3 months, 61 sticks were recovered containing 71 corals, mostly in the holes. After 1 year, three corals were confirmed to be growing well and extending outside the holes of the three sticks out of 70 sticks left in the water at the lagoon site. They survived two potentially lethal conditions, that is, high water temperatures with associated extensive coral bleaching and continuous grazing pressure from predators. This procedure is applicable for large-scale coral transplantation, not only in Japan but also in other tropical countries.     

Onset of individual growth difference in larviculture of Pacific bluefin tuna <Emphasis Type="Italic">Thunnus orientalis</Emphasis> using fertilized eggs obtained from one female

We conducted a rearing experiment on Pacific bluefin tuna (PBT) larvae, which originated from one female broodstock, and determined the growth history of the larvae to clarify when the growth difference occurs. We investigated the otolith microstructure of the PBT larvae to examine the individual growth history and to predict the age and body size at the onset of individual growth difference. Since total length (TL) of fish can be back-calculated from otolith radius, we back-calculated TLs of 100 fish of 19 days post hatch (dph) divided into three size groups (small, intermediate, large). Growth difference was recognized from 3 dph (mouth opening), and the difference became larger thereafter. Growth of large-size fish was assumed to be promoted by the feeding conditions of PBT larvae.     

Isolation of microsatellite markers by in silico screening implicated for genetic linkage mapping in Japanese pufferfish Takifugu rubripes

ABSTRACT:   The search for dinucleotide repeat microsatellites within scaffolds 1–25 of genome database JGI Fugu v3.0 for the pufferfish Takifugu rubripes revealed that 80% of microsatellite loci consisted of five to 13-fold repeats with locus-specific differences in density. Eleven out of 15 microsatellite loci isolated from the database with which genotyping using wild pufferfish was successfully performed showed polymorphism; that is, the means of the number of alleles and expected and observed heterozygosities at these 11 loci were 21.8, 0.915 and 0.829, respectively. It was confirmed that eight out of the 11 polymorphic loci were inherited through the Mendelian law and one pair of microsatellite loci derived from the same scaffold was linked. These results demonstrated that these loci are useful for constructing a linkage map in the pufferfish as DNA markers.     

Degenerative changes of the cranial cruciate ligament harvested from dogs with cranial cruciate ligament rupture

Degenerative cranial cruciate ligament (CCL) rupture is characterized histologically by degenerating extracellular matrix (ECM) and chondroid metaplasia. Here, we describe the progression of chondroid metaplasia and the changes in the expression of ECM components in canine CCL rupture (CCLR). CCLs from 26 stifle joints with CCLR (CCLR group) and normal CCLs from 12 young beagles (control group) were examined histologically and immunohistochemically for expression of type I (COLI), type II (COLII), type III collagen (COLIII) and Sry-type HMG box 9 (SOX9). Cell density and morphology of CCLs were quantified using hematoxylin–eosin staining. The percentage of round cells was higher in the CCLR group than in controls. COLI-positive areas were seen extensively in the connecting fibers, but weakly represented in the cytoplasm of normal CCLs. In the CCLR group, there were fewer COLI-positive areas, but many COLI-positive cells. The percentages of COLII-, COLIII- and SOX9-positive cells were higher in the CCLR group than in controls. The number of spindle cells with perinuclear halo was high in the CCLR group, and most of these cells were SOX9-positive. Deposition of COLI, the main ECM component of ligaments, decreased with increased COLIII expression in degenerated CCL tissue, which shows that the deposition of the ECM is changed in CCLR. On the contrary, expression of SOX9 increased, which may contribute to the synthesis of cartilage matrix. The expression of COLII and SOX9 in ligamentocytes showed that these cells tend to differentiate into chondrocytes.