Effect of dietary taurine enhancement on growth and development in red sea bream Pagrus major larvae

This study investigated the effect of feedings taurine‐enriched rotifers on the growth and development of larval red sea bream (RSB). Rotifers incubated in taurine‐enriched water at a taurine concentration of 800 mg L^?1 (T‐800) and 0 mg L^?1 (T‐0) were fed to larvae from 3 to 20 days after hatching (DAH). Notochord length, body weight and specific growth rate of T‐800 group were significantly greater than those of T‐0 at 14, 17, 9–11 and 18–20 DAH. Taurine content of larvae in the T‐800 group increased rapidly from 11 DAH and thereafter remained significantly higher than T‐0. Flexion larvae firstly appeared in both groups at 8 DAH, however, at 20 DAH post‐flexion larvae were significantly more abundant in T‐800 than T‐0. While nucleic acid and protein contents (μg mg^?1 wet fish) showed remarkable changes, ontogenetic growth in RSB larvae stage was observed to switch from hyperplastic growth to hypertrophic growth with the start of the flexion stage. Although a similar change in nucleic acid contents was observed between the two groups, the protein content (μg fish^?1) and protein/DNA ratio of T‐800 remained higher than that of T‐0 during the hypertrophic growth period. These results suggest that dietary taurine accelerates the growth and development in RSB larvae especially during hypertrophic growth (flexion stage) after the early hyperplastic growth.     

Quality evaluation of different types of non-fish meal diets for yellowtail

SUMMARY: Two feeding experiments were conducted to evaluate the feed quality of non-fish meal diets having the same protein ingredient composition but prepared as different types, and to determine the supplemental effect of crystalline essential amino acids (EAA) on feed utilization by young yellowtail, Seriola quinqueradiata . Non-fish meal diets formulated with soy protein concentrate, defatted soybean meal, corn gluten meal, meat meal, and krill meal were prepared as either soft dry pellets (SDP) or extruded pellets (EP) by using a large- or a small-sized twin screw extruder under different preparation conditions; or as a single moist pellet (SMP), each with and without EAA mixtures. Commercial yellowtail SDP was used as the control diet. Fish weighing 134 g and 237 g on average were reared with the experimental diets, for 93 (net cages) and 44 (aquariums) days, respectively. The fish fed both the control and test diets were found to have a good appetite. Growth rate and feed gain ratio were highest in the control diet group. The physiological condition of fish fed the control diet was evaluated as superior compared to those on the non-fish meal diets. Among the non-fish meal diet groups, the best performances were obtained for fish fed the SDP type diet with EAA supplement, and performance parameters excelled in the order of SDP, EP and SMP both among the diets with and without supplemental EAA. This suggests that the nutritional quality of non-fish meal diet was affected by the diet preparation method. It also indicates that supplementation of EAA could improve the quality of non-fish meal diets, irrespective of the diet type, probably as a result from the enhancement of feed protein utilization.     

Genotyping‐by‐sequencing for construction of a new genetic linkage map and QTL analysis of growth‐related traits in Pacific bluefin tuna

Pacific bluefin tuna (Thunnus orientalis) has high market value, but its wild populations have decreased in recent years. The broodstock of Pacific bluefin tuna that were hatched artificially and reared under aquaculture conditions is beginning to be used for production. The creation of broodstock with commercially valuable traits, such as rapid growth, is therefore of great interest. Genetic linkage map‐based identification of markers associated with quantitative trait loci (QTLs) facilitates marker‐assisted selection (MAS) breeding and allows efficient genetic improvement of broodstock. Single nucleotide polymorphism (SNP)‐based genetic linkage map construction using the genotyping‐by‐sequencing method can expand the number of mapped markers and help identify growth‐related QTLs. In this study, we constructed sex‐specific maps for 24 linkage groups consisting of 677 SNP and 651 microsatellite markers. The total lengths of 93 progenies in the mapping population followed normal distribution, with an average length of 9.4 mm. We performed composite interval mapping in the mapping population. QTL analysis revealed one significant QTL in LG10 on the female linkage map. The genetic linkage map—the second such map generated for Pacific bluefin tuna—and the growth‐related QTLs detected in this study will be useful for tuna aquaculture MAS programs.     

Characterization of a Phytophthora mating hormone

Water molds of the genus Phytophthora include many plant pathogens responsible for epidemics such as potato blight and sudden oak death, causing global economic damages. Sexual reproduction is of biological importance in Phytophthora and has been believed to be stimulated by unknown endogenous factors named a hormones. We describe here the chemical characterization of a Phytophthora mating hormone, a1, which was obtained from approximately 2 tons of culture fluid of one mating type of a species and which induced sexual spores on the counter-mating type at a nanogram level.     

Effect of fusion/activation protocol on in vitro development of porcine nuclear transfer embryos constructed with foreign gene-transfected fetal fibroblasts

The effect of fusion/activation protocol on in vitro development of porcine nuclear transfer (NT) embryos constructed with foreign gene-transfected somatic cells were investigated. NT embryos were produced by using enucleated M II oocytes and enhanced green fluorescence protein (EGFP) gene-transfected or non-transfected porcine fetal fibroblasts. One group of NT embryos received a single electrical pulse to induce fusion and activation simultaneously (FAS). The other group was fused 2 hr before activation (FBA) using two kinds of electrical pulses. Electrically activated NT embryos in both groups were treated with cycloheximide (CHX) before culture to assess the development to the blastocyst stage. After 6 days of culture, all morulae and blastocysts derived from EGFP-transfected fibroblasts emitted green fluorescence without mosaicism, and EGFP-gene product was also detected in all morulae and blastocysts examined. NT embryos undergoing FAS showed higher developmental capacity to blastocysts than those undergoing FBA, regardless of the EGFP transfection into the nuclear donor cells. The results also indicated that EGFP-gene transfection into nuclear donor cells has no obvious deleterious effect on the development of NT embryos to blastocysts.     

Functional insulin‐like factor 3 (INSL3) hormone‐receptor system in the testes and spermatozoa of domestic ruminants and its potential as a predictor of sire fertility

Insulin‐like factor 3 (INSL3) is essential for fetal testis descent, and has been implicated in the testicular and sperm functions in adult males; however, similar functions in domestic ruminants remain largely unknown. This study investigated the functional INSL3 hormone‐receptor system in adult ruminant testes and spermatozoa, and explored its potential to diagnose the fertility of sires. Testes and spermatozoa were obtained from fertile bulls, rams and he‐goats, whereas subfertile testes and spermatozoa were obtained only from bulls. As expected, INSL3 was visualized in Leydig cells, while we clearly demonstrated that the functional receptor, relaxin family peptide receptor 2 (RXFP2), enabling INSL3 to bind was identified in testicular germ cells and in the sperm equatorial segment of bulls, rams and he‐goats. In comparison to fertile bulls, the percentage of INSL3‐ and RXFP2‐expressing cells and their expression levels per cell were significantly reduced in the testes of subfertile bulls. In addition, the population of INSL3‐binding spermatozoa was also significantly reduced in the semen of subfertile bulls. These results provide evidence for a functional INSL3 hormone‐receptor system operating in ruminant testes and spermatozoa, and its potential to predict subfertility in sires.     

Phylogenetic analysis of the hemagglutinin (H) gene of canine distemper viruses isolated from wild masked palm civets (Paguma larvata)

Hemagglutinin (H) gene of two CDV isolates, the Haku93 and Haku00 strains, from masked palm civets was molecularly analyzed. H genes of both two CDVs contained one open reading frame encoding 607 amino acids. Nucleotide and predicted amino acid sequences of H gene of the CDV Haku93 and Haku00 revealed high similarity to those of recent field isolates such as the Yanaka and Tanu96, while they showed limited identity to those of old vaccine strains. Potential N-linked glycosylation sites in both Haku93 and Haku00 were identical to other recent CDV isolates. Phylogenetic analysis revealed that the CDV strains derived from masked palm civets were classified into the group of recent Japanese CDV isolates.     

Production of offspring from one-day-old oocytes stored at room temperature

To determine the feasibility of preserving oocytes without freezing, we stored mouse oocytes in several media at different temperatures for one day. Confocal microscopy of the metaphase-II spindle in these stored oocytes revealed gross abnormalities in both the spindle and the arrangement of chromosomes. The abnormal spindles could not be rescued by transplanting the aged spindle-chromosome complex into a fresh enucleated oocyte. A diploid parthenogenetic development showed that some of the oocytes stored at room temperature could still develop into blastocysts (10-57%). However, oocytes stored in a refrigerator (5%) or incubator (0%) lost the potential almost entirely. Fertilization of room-temperature-preserved oocytes with fresh spermatozoa by ICSI or IVF resulted in, respectively, 4 and 10%, full-term births. These results suggest that when oocytes are stored at room temperature for one day, most have irreversible damage not only to their cytoplasm but also to the spindle. However, since at least a few percent of stored oocytes retained the potential for full-term development, it may be possible to overcome these problems and develop a simple method for preserving mammalian oocytes without freezing.     

Assessment of long-term changes in nitrogen pollution load potential from sewage treatment water in the Tedori River Alluvial Fan Area, Japan

Long-term changes (1974–2007) in the nitrogen pollution load potential (NPLP) arising from sewage treatment water were assessed in the Tedori River Alluvial Fan Area of Japan. The total NPLP from sewage treatment systems (STS) during the 34 year period was 439 t (10³ kg) year⁻¹ from about 260,000 users in 1974 increasing to a peak of 793 t year⁻¹ in 1992 from about 363,000 users, and then decreasing to 676 t year⁻¹ from about 400,000 users in 2007. The NPLP outflow into the area increased from 356 t year⁻¹ in 1974 to a peak of 596 t year⁻¹ in 1985 followed by a rapid decrease to 98 t year⁻¹ in 2007. The NPLP outflow from the public STS to the Japan Sea began in 1979 and rapidly increased to 575 t year⁻¹ in 2007 from about 362,000 users. This represents 85.5% of the total NPLP. The NPLP from septic tanks in the area was 356 t year⁻¹ from about 107,000 users in 1974 gradually increasing to a peak of 587 t year⁻¹ from about 177,000 users in 1985 before rapidly decreasing to 60 t year⁻¹ from about 15,000 users in 2007. Although the current NPLP is about 98 t year⁻¹ in the study area, the average NPLP during 34 years was very different at 424 t year⁻¹. NPLP assessments affecting groundwater and closed water bodies should consider long-term processes of nitrogen pollution from STS over time periods compatible with the residence time of the receiving waters.     

Pyridinoline concentrations in muscular and skin collagen of fish and relationship between collagen solubility and pyridinoline concentration in fish muscular collagen

Pyridinoline (Pyr), one of the mature crosslinks of collagen, was determined in muscular collagen of three species of fish. The amounts of muscular Pyr in red sea bream, yellowtail, and tiger puffer were 3.4, 8.8, and 50.3 mmol/mol collagen, respectively, indicating that the Pyr concentration in muscular collagen differs greatly among fish species. The Pyr concentration in tiger puffer muscular collagen was the greatest, but it was only one-fourth that in rabbit muscle. As in mammalian skin collagen, Pyr was not detected in skin collagen of red sea bream and yellowtail. However, tiger puffer skin contained Pyr (3.75 mmol/mol collagen). The presence of Pyr would have a relationship to some features of tiger puffer skin, such as mechanical strength and thickness. Pyr concentrations in acid-soluble collagen (ASC), pepsin-solubilized collagen (PSC), and insoluble collagen (ISC) in muscles of three species of fish were determined. Pyr was found in ISC > PSC > ASC, from the highest to the lowest concentration, and the concentration in ISC was 45–200 times that in ASC. Therefore, Pyr crosslinks that are formed between collagen molecules would have a close relationship to collagen solubility.