Microsatellite marker isolation and cultured strain identification in <Emphasis Type="Italic">Carassius auratus gibelio</Emphasis>

Microsatellites have become the preferred molecular markers for strain selection and genetic breeding in fish. In this study a total of 105 microsatellites were isolated and identified in gibel carp (Carassius auratus gibelio) by microsatellite sequence searches in GenBank and other databases and by screening and sequencing of positive clones from the genomic library enriched for AG and GATA repeats. Moreover, nineteen microsatellites were randomly selected to design locus-specific primer pairs, and these were successfully used to identify and discriminate different cultured strains of gibel carp including strains A, D, L, and F. Three different types of microsatellite pattern were distinguished by the number and length of fragments amplified from the 19 primer pairs, and some microsatellite primer pairs were found to produce different microsatellite patterns among strains and strain-specific fragments. In addition, some duplicated alleles were also detected in two microsatellite patterns. Therefore, the current study provides direct molecular markers to discriminate among different cultured strains for selective breeding and aquaculture practice of gibel carp.     

Effect of N-acetyl cysteine and glycine supplementation on growth performance,glutathione synthesis,and antioxidative ability of grass carp, <Emphasis Type="Italic">Ctenopharyngodon idella</Emphasis>

An 8-week feeding trial was conducted to evaluate the effect of N-acetyl cysteine (NAC) and glycine supplementation on growth performance, glutathione (GSH) synthesis, and antioxidative ability of grass carp, Ctenopharyngodon idella. Four practical diets were formulated: control, control + 0.2% NAC, control + 0.5% glycine, and control + 0.2% NAC + 0.5% glycine. Each diet was randomly assigned to quadruplicate groups of 30 fish (approximately 8.8 g). Weight gain and specific growth rate were significantly increased with the supplementation of NAC. Supplementation of NAC plus glycine significantly increased the feed efficiency. Glutathione peroxidase (GPx) and γ-glutamine cysteine synthase (γ-GCS) in plasma were significantly increased with the supplementation of NAC plus glycine. GSH in plasma increased and malondialdehyde (MDA) decreased in fish fed diets supplemented with NAC. Respiratory burst, superoxide dismutase (SOD), and catalase (CAT) activity were not affected by NAC or glycine. These results clearly indicated that NAC improved the growth performance and restored GSH of grass carp, supplemented NAC together with glycine enhanced GSH synthesis, and improved the antioxidative ability of grass carp.     

The development of the Norwegian wrasse fishery and the use of wrasses as cleaner fish in the salmon aquaculture industry

Norway leads the world aquaculture production of Atlantic salmon Salmo salar and farmed Norwegian Atlantic salmon is currently consumed around the globe. However, sea lice infestation is a major problem faced by the salmon aquaculture industry in Norway and elsewhere. The use of wild-caught cleaner fish, mainly wrasses, has been recommended over the other available methods as the most economical and environmentally friendly option to control sea lice infestation in salmon farming. Here, we review the development of the Norwegian wrasse fishery and the use of wrasses as cleaner fish. In this document, we address the sea lice problem and introduce the main wrasse species employed as cleaner fish, document the cleaning behaviour of wrasses, present the development of a new wrasse fishery associated with the salmon aquaculture industry, and finally, we identify the main challenges associated with the intensive use of wild-caught cleaner wrasses and provide some insight for future directions of the wrasse fishery and further development of aquaculture techniques to supply salmon facilities with domesticated cleaner fish.     

Effects of light intensity on molting,growth, precocity,digestive enzyme activity,and chemical composition of juvenile Chinese mitten crab <Emphasis Type="Italic">Eriocheir sinensis</Emphasis>

A 74-day growth trial was carried out to investigate the effects of light intensity on the juvenile Chinese mitten crab, Eriocheir sinensis, under semi-natural conditions. The experiment had three light intensity treatment groups, natural light (NL), middle light (ML), and low light (LL), as light intensity became weaker. The results indicated that light intensity had no significant effect on molting interval and molting frequency but did have a significant effect on the molting weight gain of the crab. Molting weight gain in group NL was significantly higher than that in group LL (P < 0.05). Specific growth rate in weight (SGRw) and in carapace width (SGRcw), weight gain, and final body weight were significantly affected by light intensity (P < 0.05). No significant difference was detected in survival between groups (P > 0.05). The precocious rate in groups NL, ML, and LL was 26.14, 15.48, and 17.14%, respectively. The precocious rate in group NL was significantly higher than that in groups ML and LL (P < 0.05). Chemical composition of the crab body was significantly affected by light intensity, but the activity of alkaline phosphatase, trypsin, and pancreatic lipase was not significantly affected by light intensity. The results indicated that the submitted light intensity was useful in reducing the precocious rate without affecting the normal growth of juvenile E. sinensis.     

Inhibition of hirame rhabdovirus growth by RNA aptamers

RNA aptamers are artificial nucleic acids that specifically bind to a wide variety of targets. They are an effective tool for pharmaceutical research and development of antiviral agents. Here, we describe four Hirame rhabdovirus (HIRRV)‐RNA aptamers (H1, H2, H3 and H4) that we obtained from an in vitro process called the systematic evolution of ligands by exponential enrichment (SELEX). The HIRRV‐RNA aptamers specifically bind to HIRRV. Hirame natural embryo (HINAE) cells treated with virus and the RNA aptamer showed a decrease in appearance of cytopathic effect when compared with control (treated only with virus). Rhodovulum sulfidophilum was transformed with genes for the RNA aptamers, and the aptamers were detected in the culture medium, indicating that they were secreted from the cells. Thus, the recombinant R. sulfidophilum might be a powerful tool for the prevention of HIRRV in aquaculture.     

Species identification method for <Emphasis Type="Italic">Scombrops boops</Emphasis> and <Emphasis Type="Italic">Scombrops gilberti</Emphasis> based on polymerase chain reaction-restriction fragment length polymorphism analysis of mitochondrial DNA

ABSTRACT:   Gnomefish Scombrops boops and Scombrops gilberti are commercially important fishes in Japan, but these species are often confused in the markets because of their morphological similarity. To identify these two species, we performed nucleotide sequencing and restriction fragment length polymorphism (RFLP) analysis on 16S ribosomal RNA (rRNA) gene and the control region in mitochondrial DNA. Five and 12 nucleotide substitutions were observed between species in the 777-bp 16S rRNA gene and 471-bp control region, respectively. Diagnostic restriction sites for discriminating between S. boops and S. gilberti were found in the 16S rRNA gene, but not in the control region. Polymerase chain reaction (PCR)–RFLP analysis using two enzymes, Eco NI and Mva I, clearly discriminated between S. boops and S. gilberti identified by meristic characters. The PCR–RFLP analysis identified most of the 168 Scombrops young caught in the coastal waters of the Izu and Miura peninsulas as S. boops , suggesting that S. gilberti juveniles are rare in this area.     

Sex reversal and growth performance in juvenile females of the freshwater crayfish <Emphasis Type="Italic">Cherax quadricarinatus</Emphasis> (Parastacidae): effect of increasing temperature and androgenic gland extract in the diet

Monosex male culture of C. quadricarinatus is proposed for increasing yields because males reach a larger size at harvest than females. An experiment was done to evaluate the effects of androgenic gland extract and temperature on sex reversal and somatic growth in early juvenile females. Females were exposed to the following experimental conditions: C: control food and temperature maintained at 26 ± 1°C; HT (high temperature): control food and temperature maintained at 28.5 ± 1°C; VHT (very high temperature): control food and temperature maintained at 31 ± 1°C; AG: food with enriched with androgenic gland (1/10 dose for each juvenile per day) and temperature maintained at (26 ± 1)°C. The juveniles were weighed, sexed, and growth increment (GI) and growth rate (GR) were calculated twice a month. The HT and AG groups differed from the C group in GI and GR indicating a greater somatic growth. At the end of the experiment, the HT and AG groups had similar weight but only the HT group had enhanced oocyte diameter, with some vitellogenic oocytes compared to the C group. In the HT group, there was significant sex reversal, as indicated by development of male and intersex male characteristics in different individuals. Both temperature and AG diet have a high potential for culture of this species because of their enhancement of somatic growth; higher temperature increases the proportion of males, a desirable characteristic of cultured populations.     

Ontogenetic development of digestive enzymes and effect of starvation in miiuy croaker <Emphasis Type="Italic">Miichthys miiuy</Emphasis> larvae

The ontogenetic development of the digestive enzymes amylase, lipase, trypsin, and alkaline phosphatase and the effect of starvation in miiuy croaker Miichthys miiuy larvae were studied. The activities of these enzymes were detected prior to exogenous feeding, but their developmental patterns differed remarkably. Trypsin activity continuously increased from 2 days after hatching (dah), peaked on 20 dah, and decreased to 25 dah at weaning. Alkaline phosphatase activity oscillated at low levels within a small range after the first feeding on 3 dah. In contrast, amylase and lipase activities followed the general developmental pattern that has been characterized in fish larvae, with a succession of increases or decreases. Amylase, lipase, and trypsin activities generally started to increase or decrease at transitions from endogenous to exogenous feeding or diet changes, suggesting that these enzymatic activities can be modulated by feeding modes. The activities of all the enzymes remained stable from 25 dah onwards, coinciding with the formation of gastric glands and pyloric caecum. These results imply that specific activities of these enzymes underwent changes due to morphological and physiological modifications or diet shift during larval development but that they became stable after the development of the digestive organs and associated glands was fully completed and the organs/glands functioned. Trypsin and alkaline phosphatase were more sensitive to starvation than amylase and lipase because delayed feeding up to 2 days after mouth opening was able to adversely affect their activities. Enzyme activities did not significantly differ among feeding groups during endogenous feeding; however, all activities were remarkably reduced when delayed feeding was within 3 days after mouth opening. Initiation of larvae feeding should occur within 2 days after mouth opening so that good growth and survival can be obtained in the culture.     

Compensatory growth and body composition of juvenile black rockfish <Emphasis Type="Italic">Sebastes schlegeli</Emphasis> following feed deprivation

ABSTRACT:   Compensatory growth, feeding rate, feed efficiency and chemical composition of juvenile black rockfish (mean weight 1.43 g) were investigated for 35 days after a 14-day feed deprivation treatment under four feeding conditions: one group continuously fed (control) and the other three groups fasted for 5 days (F5), 10 days (F10) and 14 days (F14). All fasted fish were re-fed from day 15. Only F5 achieved the same body weight as the control, indicating that complete compensation occurred in F5. The specific growth rate (SGR) of F5 was the highest at day 21 and then decreased thereafter, showing higher values than the control at days 21, 28 and 42. In contrast, although SGRs of F10 and F14 were higher than that of the control during the whole refeeding period except day 21, they did not catch up the control in body mass, indicating that only partial compensation occurred in F10 and F14. The feeding rate (FR) of all groups except F14 changed in a pattern similar to SGR (Spearman's rank correlation, r _s > 0.9), suggesting that SGR varied depending on FR. Similar feeding efficiencies (FEs) were found in the four groups and they did not vary significantly during the whole refeeding period, suggesting that FE was not the factor affecting SGR. At day 14, the ratios of lipid to lean body mass in F10 and F14 were lower than those in the control and F5, and there was no difference between the control and F5. At day 49, however, only F14 showed a lower value than the other three groups, and there was no difference among the three groups. These results indicate that juvenile black rockfish fasted for 5–14 days can exhibit compensatory growth after refeeding, but timing and degree vary depending on the duration of feed deprivation.