Effect of Illuminance Transition Rate on Survival of Juvenile Pacific Bluefin Tuna,Thunnus orientalis

To investigate the reason for the high mortality of cultured juvenile Pacific bluefin tuna (PBT), especially during land‐based culture, we examined the effect of the rate of light‐intensity change during the dawn period. Juvenile PBT kept in a natural light environment (control) were compared with those exposed to an artificially slowed change in light intensity during the dawn period (test group). For the test group, lamps, which were connected to automatic timers and placed above the rearing tank, were switched on from 0345 to 0515 h at 15‐min intervals. After 9 d, the survival rate of the test and control group was identical. Between Day 1 and Day 6, whole‐body cortisol levels were not different between groups. However, whole‐body glucose levels in the test group were significantly higher than those in the control group. Both groups displayed a diel rhythm in plasma cortisol concentration that peaked at 0300–0600 h. These results suggest that slowing the increase in light intensity during the dawn period did not improve survival and that low light intensity itself induces high mortality in cultured PBT juveniles.     

Variations in the morphology of Rhizomucor pusillus in granulomatous lesions of a Magellanic penguin (Spheniscus magellanicus)

This report presents a new case of mucormycosis encountered in penguin characterized by morphological variation of hyphae and presence of sporangia with numerous sporangiospores. A 4.5-year-old Magellanic penguin (Spheniscus magellanicus) died after exhibiting anorexia, poor nutritional condition and dyspnea. Multiple nodular lesions were observed in the thoracic and abdominal regions. Histopathologically, hyphae of various sizes were seen in the lungs, air sac and nodular lesions. Myriad sporangiospores and several sporangia were observed in/around the bronchi or parabronchi. The very narrow and short hyphae in the nodules were not consistent with the characteristics of Mucorales. However, for most hyphae, including those in the nodules, sporangiospores and sporangia, immunohistochemistry revealed Mucorales-positive reactions. In addition, these fungi were identified as Rhizomucor pusillus by gene analysis.     

<Emphasis Type="Italic">In silico</Emphasis> genomic subtraction guides development of highly accurate,DNA-based diagnostics for <Emphasis Type="Italic">Ralstonia solanacearum</Emphasis> race 3 biovar 2 and blood disease bacterium

New rapid diagnostic methods are urgently needed to discriminate the quarantine pathogen Ralstonia solanacearum (Rs) race 3 biovar 2 (R3B2) from other populations of Rs that lack the adaptation to cause bacterial wilt disease in temperate regions. We used an in silico bioinformatic approach to identify several genome sequences potentially specific to R3B2 strains. Primer sets were designed to PCR-amplify sequences in these regions, and four sets were ultimately shown to be >99% accurate for detection of R3B2 strains. On the basis of these results, several primers were designed to enable development of a loop-mediated isothermal amplification assay that was rapid, technologically simple, and essentially 100% accurate for identification of R3B2 when applied to a comprehensive collection of geographically diverse Rs strains. We fortuitously found that a sequence in one of the “R3B2-specific” regions has ~90% identity to a sequence present in strains of the blood disease bacterium (BDB), a member of the Rs species complex that infects banana. Alignments of these sequences allowed design of a second PCR primer set that proved 100% accurate for identification of BDB strains when tested on the 22 BDB strains available to us. These results demonstrate the power of in silico genomic subtraction for rapid identification of population-specific DNA sequences and for the development of simple, reliable detection methods for Rs subpopulations.     

A Comparison of the Effects of Two Prey Enrichment Media on Growth and Survival of Pacific Bluefin Tuna,Thunnus orientalis,Larvae

Three experiments were conducted to investigate the growth, survival, and standardized cohort biomass of Pacific bluefin tuna, Thunnus orientalis, larvae fed nutritionally enhanced prey during the first week of feeding using two commonly used, commercially available enrichment media, AlgaMac Enrich and Marine Glos. T. orientalis larvae exhibited exponential growth in standard length and dry weight. The daily specific growth rates in length and weight are the first reported for T. orientalis larvae and the averages ranged from 3.8 to 4.1% and 27.5%, respectively, for larvae in the AlgaMac treatment and from 4.1 to 6.1% and 31.5%, respectively, in the Marine Glos treatment. Average daily growth rates in length ranged from 0.16 to 0.23 mm/d for larvae in the AlgaMac treatment and from 0.17 to 0.27 mm/d for those in the Marine Glos treatment. Daily growth rates in length were similar to those reported for other tuna larvae reared in the laboratory but slower than most published estimates for larval tunas in situ at similar water temperatures. Mean prey number per gut was positively associated with mean prey level in the tank. Both enrichment media appear to be good sources of nutritional improvement of planktonic prey for T. orientalis larvae.     

Gonadal morphogenesis and sex differentiation in cultured chub mackerel,Scomber japonicus

The timing of primordial germ‐cell (PGC) migration with regard to the gonadal anlagen, gonad formation and sex differentiation was examined histologically in the chub mackerel (Scomber japonicus) at 5–190 days post hatching (dph). At 5 dph, PGCs appeared on the peritoneal epithelium surface or in the mesentery, on the dorsal side of the abdominal cavity. By 10 dph, stromal cells around the PGCs proliferated. The gonadal primordium was formed by 15 dph. The gonadosomatic index was 0.01% at 30 dph and increased thereafter (0.32% in females and 0.04% in males at 160 dph). Ovarian differentiation occurred at 30–40 dph, indicated by ovarian cavity formation (elongation and fusion of the upper and lower ovarian edges). Meiosis was subsequently initiated. A few meiotic oocytes surrounded the cavity at 50 dph; most were in the perinucleolus stage at 60 dph and attained a diameter of 60–70 μm at 190 dph. Testicular differentiation occurred at 30 dph, indicated by the formation of the sperm duct primordium. Spermatogonia gradually proliferated, developing into spermatocytes at the chromatin–nucleolus stage (after 90 dph) and subsequently into spermatids and spermatozoa (160 dph). These data could aid the development of seeding and cell‐engineering technologies for scombrid fish.     

拖拉机在牧草地上自动引导行走的控制

利用车辆运动学模型,应用最优控制理论,对拖拉机自动引导行走方法进行了研究。利用前馈控制方法生成了车辆自动行走的轨道;通过对车辆运动状态方程的线性化,设计了车辆沿生成的轨道引导行走的负反馈控制器。最后利用该方法在牧草地上进行了实车实验,结果表明:拖拉机在倾斜度小于3°的地面上直线行走时的横向标准偏差小于0.08m。     

Evaluation of genetic variation among wild rice populations in Cambodia

A total of 448 samples in five natural populations of wild rice (Oryza rufipogon) were collected in Cambodia. They were examined using 12 SSR and two chloroplast markers to evaluate the degree of variation among populations and the genetic structure within populations. In the two annual populations, the number of plants with homozygous alleles at all 12 SSR loci were high (66.3% and 79.5%), suggesting that these plants propagate mainly through self-pollination. In the three perennial populations, no individuals had all homozygous genotypes, but redundant genotypes resulted from clonal propagation were observed. Percentages of the redundant genotypes were highly varied (3.6%, 29.2% and 86.0%). This may be due to the different stable levels of environmental conditions. As for chloroplast genome, most of the wild plants showed the same chloroplast types as most Indica-type cultivars have. However, plants with different chloroplast types were maintained, even in the same population. In tropical Asian countries, many wild rice populations were observed under similar ecological conditions examined in this study. Therefore, the present results concerning population structure will be important to further elucidate genetic features of wild rice, and will also give strong clues to utilize and conserve wild natural genetic resources.     

Long-term monitoring of feral genetically modified herbicide-tolerant Brassica napus populations around unloading Japanese ports

Genetically modified, herbicide-tolerant (GMHT) Brassica napus plants originating from seed spill have recently been found along roadsides leading from Japanese ports that unload oilseed rape. Such introductions have potential biodiversity effects (as defined by the Cartagena Protocol): these include replacement of native elements in the biota through competitive suppression or hybridization. We conducted surveys in the period 2006–2011 to assess such threats. We examined shifts in the population distribution and occurrence of GMHT plants in 1,029 volunteer introduced assemblages of B. napus, 1,169 of B. juncea, and 184 of B. rapa around 12 ports. GMHT B. napus was found around 10 of 12 ports, but its proportion in the populations varied greatly by year and location. Over the survey period, the distributions of a pure non-GMHT population around Tobata and a pure GMHT population around Hakata increased significantly. However, there was no common trend of population expansion or contraction around the 12 ports. Furthermore, we found no herbicide tolerant B. juncea and B. rapa plants derived from crosses with GMHT B. napus. Therefore, GMHT B. napus is not invading native vegetation surrounding its populations and not likely to cross with congeners in Japanese environment.