Production of androgenetic amago salmon<Emphasis Type="Italic"> Oncorhynchus masou ishikawae</Emphasis> with dispermy fertilization

With the aim of improving artificial androgenesis in teleost fishes, we tested two methods for producing androgenetic diploids of amago salmon (Oncorhynchus masou ishikawae), namely, fertilization of gamma-ray irradiated eggs with fused spermatozoa (sperm-fusion method) and the fertilization of irradiated eggs with untreated sperm followed by the blocking of cell division (mitosis-inhibition method). Our results showed that the optimal condition for sperm fusion was to treat the sperm with 50% polyethylene glycol (molecular weight 7500) for 100 s. The efficiency of the two methods of androgenesis was compared in terms of fertilization rate, hatching rate, and larval survival after hatching. The rate of fertilization was lower with the sperm-fusion method than with the mitosis-inhibition method, but the reverse was true for the hatching rate. The survival rate of hatched larvae was the same with the two methods. Androgenesis was confirmed with a recessive albino color marker, and all viable offspring were found to be heterozygous based on analysis of the microsatellite markers. Our results suggest that androgenesis with the sperm-fusion method is a promising approach with potential applications in both aquaculture breeding programs and the preservation of endangered freshwater fishes.     

Nutritional evaluation of rotifers in rearing tanks without water exchange during seed production of amberjack <Emphasis Type="Italic">Seriola dumerili</Emphasis>

We evaluated the nutrient contents of rotifers sampled from larval-rearing tanks (tank rotifers) without water exchange during the seed production of amberjack Seriola dumerili at three facilities (Kamiura, Kagoshima, and Miyazaki) and compared them with the nutrient contents of freshly enriched rotifers. Compared to the enriched rotifers, the lipid contents, especially neutral lipids and proportion of 22:6n-3, tended to decrease in the tank rotifers. These trends were clearer at Miyazaki where the tank rotifers were sampled before daily supplementation of microalgae (Nannochloropsis). Crude protein content of the tank rotifers did not decrease markedly although the proportion of lysine tended to decrease. Vitamin C and E contents of the tank rotifers decreased significantly only at Miyazaki. Calcium content of the tank rotifers increased at Kamiura and Miyazaki, and the increases in iron and manganese contents of the tank rotifers at Miyazaki and zinc content at Kagoshima were pronounced. These results suggest that the nutritional value of rotifers in larval-rearing tanks without water exchange can be maintained by appropriate supplementation of microalgae. The effect of certain minerals that became high in tank rotifers on subsequent larval development requires further investigation.     

Influence of soya saponin, soya lectin, and cholyltaurine supplemented to a casein-based semipurified diet on intestinal morphology and biliary bile status in fingerling rainbow trout Oncorhynchus mykiss

To investigate the cause of the changes in intestinal morphology and biliary bile status of rainbow trout Oncorhynchus mykiss fed defatted soybean meal (SBM)-based diets, casein-based semipurified diets supplemented with soya saponin, soya lectin, and cholyltaurine were fed to rainbow trout for 6 weeks. An unsupplemented control diet and a SBM-based diet were also tested as references. Poor development of microvilli and pinocytotic vacuoles, and accumulation of large vacuoles in the epithelial cells were observed in the distal intestine of fish fed diets containing saponin but not cholyltaurine. Hyperplastic connective tissue in the mucosal folds of the distal intestine was observed in fish fed a diet containing both saponin and lectin but not cholyltaurine. However, intestinal histological features in fish fed diet supplemented with cholyltaurine and lectin and/or saponin were similar to those in the control diet group. Liver morphology and biliary bile status were not affected by saponin and lectin. These results suggest that the abnormal features of the distal intestine of rainbow trout fed SBM-based diets are caused by the combination of soya saponin and soya lectin, and that supplemental cholyltaurine plays certain roles in normalizing the intestinal abnormalities caused by the saponin and lectin.     

Morphological and physiological studies on densely branched lateral roots triggered by localized phosphate in Sesbania cannabina

Densely branched lateral roots (DBLRs) in Sesbania cannabina are formed in response to patchily distributed phosphorus (P) in volcanic soils. Little attention has been paid to morphological and physiological responses of DBLRs. Here, we investigated the relation between plant growth and DBLR development, enzymatic activities involved in P acquisition, and the influence of arbuscular mycorrhizal fungi (AMF), which contribute to P uptake, to clarify the function of DBLRs. We investigated DBLR development induced by localized application of P fertilizer and we compared the activities of phosphoenolpyruvate carboxylase (PEPCase) and acid phosphatase (APase) between DBLRs and non‐DBLRs. Additionally, plants were grown with or without AMF to investigate the effect of AMF colonization on the numbers of DBLRs and plant P uptake, and we compared AMF colonization between DBLRs and non‐DBLR roots. Secondary to quaternary lateral DBLRs were produced after the primary lateral roots passed near P fertilizer. P_i content per DBLR increased as DBLRs developed, promoting higher shoot growth. Under P deficiency, PEPCase and APase activities increased in non‐DBLR, but were significantly lower in DBLRs in the same plants. AMF inoculation changed the root system architecture by significantly decreasing the number of DBLRs, and AMF colonization was lower in DBLRs than in non‐DBLRs. Our results indicate that DBLR formation is a P‐coacquisition strategy of S. cannabina grown in P‐deficient andosolic soil. Roots that form DBLR are clearly different from non‐DBLR roots in morphological and biochemical response and AMF symbiosis.     

Long‐term changes in recruitment of age‐0 Pacific bluefin tuna (Thunnus orientalis) and environmental conditions around Japan

Recruitment of age‐0 Pacific bluefin tuna (Thunnus orientalis) from 1952 to 2014 was examined by a sequential regime shift detection method. The regime shifts in recruitment were detected in 1957, 1972, 1980, 1994 and 2009. The durations of regime shift ranged from 8–15 years and averaged 13.0 years. In both the total (1952–2014) and data rich (1980–2014) periods, negative relationships were found between recruitment and the Pacific Decadal Oscillation in autumn, and positive relationships were found between recruitment and sea surface temperature (SST) anomalies in the northern part of the East China Sea, in the southwestern part of the Sea of Japan, and in the waters off Shikoku and Tokai in summer and autumn. The 1994 and 2009 regime shifts in recruitment occurred in the same years as shifts in SST anomalies in the northern part of the East China Sea in summer. These results suggest that the ocean conditions in the northern part of the East China Sea are closely related to recruitment of Pacific bluefin tuna, and that the warmer conditions result in higher recruitment of the species.     

Phylogenetic study of Japanese Dickeya spp. and development of new rapid identification methods using PCR–RFLP

Forty-one representative Japanese Dickeya spp. (Erwinia chrysanthemi) strains isolated from 24 plants in Japan were investigated using multilocus sequence analysis of recA, dnaX, rpoD, gyrB and 16S rDNA; PCR–RFLP (restriction fragment length polymorphism) of recA, rpoD and gyrB genes; PCR genomic fingerprinting; and biochemical tests. Based on the recA, dnaX, rpoD, gyrB and 16S rDNA sequences and PCR genomic fingerprinting, the strains were essentially divided into six groups (I–VI). Group I corresponded to D. chrysanthemi, group II corresponded to D. dadantii, group III to D. dianthicola and group IV to D. zeae. Meanwhile, group V and group VI could not be assigned to any existing Dickeya species, and they were deduced to be two putative new species. The PCR–RFLP analysis of gyrB, rpoD and recA clearly differentiated the six groups of Dickeya strains. From the results of the biochemical tests, the strains were assigned to biovars 1, 3, 5, 8 and 9; only one strain (SUPP 2525) was not assignable to the existing biovars. We also showed that the PCR–RFLP analysis of rpoD, gyrB and recA can be used as a rapid technique to identify Japanese Dickeya strains.     

Development of an adaptive marine ecosystem management and co-management plan at the Shiretoko World Natural Heritage Site

The Marine Management Plan for the Shiretoko World Natural Heritage Site, Japan, provides a case study for adaptive marine ecosystem management and co-management of coastal fisheries. Shiretoko was the third World Natural Heritage Site registered in Japan and earned this title because of its (i) formation of seasonal sea ice at some of the lowest latitudes in the world, (ii) high biodiversity, and (iii) many globally threatened species. The natural resource management plan of the Shiretoko site is characterized by transparency and consensus building, because (i) UNESCO and IUCN require that the plan be sustainable and (ii) the Government of Japan has guaranteed local fisheries that there will be no additional regulations included in the plan. The Marine Management Plan describes which species and factors are monitored, how these data are evaluated, and how the benchmarks specified by ecosystem management are determined. The plan will provide a valuable example for the establishment of “environment-friendly fisheries” in Japan and other countries, because it includes voluntary activities by resource users that are suitable for use in a local context, flexible to ecological/social fluctuations, and efficiently implemented through increased legitimacy and compliance. This approach is appropriate for coastal communities where a large number of small-scale fishers catch a variety of species using various types of gear. We develop a method to evaluate fisheries integrity by-catch and yield data.