Optimal egg viability storage conditions in two commercial fairy shrimps (Crustacea: Branchiopoda: Anostraca) from Thailand

Optimal egg viability storage conditions for two commercial fairy shrimps, Branchinella thailandensis and Streptocephalus sirindhornae, were investigated. Eggs for each treatment were kept under eight different conditions at four different temperatures for 24 months. Every 2 months, representative eggs were hatched to evaluate hatching percentages. Low temperature and oxygen, darkness and dry conditions significantly influenced egg hatchability of both species. Undehydrated eggs hatchability in all conditions ranged from 0 to 10% in S. sirindhornae and 0 to 20% in B. thailandensis. Hatching percentages of B. thailandensis were higher than those of S. sirindhornae. Hatchability under all experimental regimes continuously decreased over time, except for dry eggs stored at ?18°C in dark‐anoxic conditions. Hatchability was markedly delayed in both species over time. We demonstrate that both species’ eggs can be stored at ?18°C at least for 24 months with high hatchability in B. thailandensis (80%) and S. sirindhornae (60%) under dry, dark, anoxic conditions.     

The influence of diet on the grass carp intestinal microbiota and bile acids

To elucidate the influence of different diet on the intestinal microbe and bile acids, we characterized the microbiota and bile acids in the hindgut content of grass carp fed on formula feed (FF group) or Sudan grass (SG group). Fusobacteria and Proteobacteria were significantly more represented in FF group than in SG group whereas Bacteroidetes was significantly more abundant in SG group than in FF group. Simpson diversity was significantly higher in FF group than in SG group (t = 2.33, P < 0.05). Chenodeoxycholic acid (CDCA) was the most abundant primary bile acid in the two groups, with average concentrations of 1.03 ± 0.62 and 4.44 ± 1.80 ng mg^?1 in SG and FF group respectively. The most abundant secondary bile acid was deoxycholic acid (DCA) in SG group and ursodeoxycholic acid (UDCA) in FF group, with average concentrations of 0.17 ± 0.06 and 2.67 ± 0.88 ng mg^?1 respectively. UDCA is significantly more abundant in FF group than in SG group, and the total bile acids were higher in FF group than in SG group. Cetobacterium and Fusobacteriaceae U114 were significantly related with the concentrations of CDCA (r = 0.85, P < 0.05 and r = 0.82, P < 0.05 respectively) and UDCA (r = 0.92, P < 0.01 and r = 0.92, P < 0.01 respectively). However, Bacteroides was negatively related with the concentration of UDCA (r = ?0.67, P < 0.05). Overall, there existed certain relationship between the intestinal microbes and the faecal bile acids, and they were both influenced by the diet.     

Two biomass preparation methods provide insights into studying microbial communities of intestinal mucosa in grass carp (Ctenopharyngodon idellus)

Animal digestive tract is habitat for a large number of autochthonous microbiota, which play central roles in multiple biological and physiological processes of the host. In this study, two different micro‐biomass preparation methods were employed to evaluate the diversity of intestinal mucosa‐associated microbiota in grass carp (Ctenopharyngodon idellus). Genomic DNAs were isolated either directly from intestinal mucosal samples (group A), or from micro‐biomass after microbial dissociation (group B). Community richness, diversity and evenness indices were all higher in group B, but differences were not statistically significant (P = 0.97, P = 0.33, P = 0.34 respectively). Furthermore, group B samples exhibited an increased ratio of bacterial DNA in comparison with group A samples, but the difference was also not statistically significant (P = 0.74). In addition, there were no statistically significant differences between the two groups (P > 0.05) at the taxonomic level. Our results support previous findings that there exists a great abundance of the intestinal mucosa‐adherent microbiota in the grass carp; among these, Proteobacteria, Firmicutes, Bacteroidetes, Actinobacteria, Spirochaetes and Fusobacteria were the most common phyla. Within these microbiota, Paenibacillus, Bacteroides, Bacillus and Cetobacterium genera comprise the majority of the community, implicating their functional importance (e.g. as probiotics) to their host. Our results contribute towards a better understanding of the intestinal microbial profile of grass carp. Both micro‐biomass preparation techniques proved to be feasible for studying mucosa‐adherent microbiota of grass carp; however, the second method (group B) provides a protocol that is somewhat more effective than the first method (group A).     

Comparative survival and growth performance of European lobster larvae,Homarus gammarus,reared on dry feed and conspecifics

A bottleneck of crustacean larval culture concerns nutrition and associated cannibalism in communal rearing systems, which impact on larval survival, development and growth. For early‐stage European lobster, Homarus gammarus larvae, feeding ecology and body composition are largely unknown. We initiated four progressive feeding experiments (novel feed types, feeding regime and feed size and cannibalism effects) on growth and survival, to inform and update husbandry protocols. Performance of larvae offered a dry commercial feed was not significantly different compared with a conventional wet plankton feed of the same ration and size grade (both within 600–1,000 μm). Further experiments found that the same ration of dry feed offered six times daily improved development and growth, over the conventional regime of three times daily. Small‐grade dry feed (particles: 250–360 μm) improved larval performance compared with a larger feed (360–650 μm). Larvae were also fed different proportions of dry feed and/or conspecifics in both communal and individual rearing systems (the latter preventing cannibalism via segregation). Individually reared larvae, fed only dead conspecifics, displayed the greatest survival (80%) to postlarvae. This underlines the impact of cannibalism on survival and nutrition in H. gammarus larviculture. A final experiment analysed H. gammarus zoea 1 composition, identifying deficiencies in ash and carbohydrate in lobster feeds. This suggests a need for a species‐specific, formulated dry feed for H. gammarus larviculture. Our research represents the first investigation of H. gammarus larval composition and dietary requirements and highlights decreased growth potential associated with providing nutrition solely from generic commercial feed.     

Effect of l‐ascorbyl‐2‐polyphosphate supplementation on growth performance,body composition,antioxidative capacity and salinity stress tolerance of juvenile Pacific white shrimp,Litopenaeus vannamei

An 8‐week feeding trial was conducted to evaluate the effects of ascorbic acid (AsA), in the form of l ‐ascorbyl‐2‐polyphosphate (LAPP) on growth performance, body composition, antioxidative capacity and salinity stress tolerance of juvenile Pacific white shrimp, Litopenaeus vannamei. Five practical diets (46% crude protein and 7.6% lipid) supplemented with graded levels of AsA (14.64, 48.55, 84.98, 308.36 and 639.27 mg kg^?1 diet) were fed to five replicate groups of L. vannamei (mean initial wet weight 0.57 g). No significant differences were found on growth performance among all treatments. However, whole body lipid content significantly decreased with dietary AsA levels increasing. Activities of total antioxidant capacity, glutathione reductase and glutathione peroxidase were significantly affected by dietary AsA levels. Shrimp fed LAPP‐free diet had higher malondialdehyde content than those fed the diets supplemented with LAPP. Dietary AsA levels higher than 308.36 mg kg^?1 diet increased the survival of shrimps after 1, 2 and 3 h of acute salinity change. Broken‐line regression analysis on survival after 3 h of salinity stress and second‐degree polynomial regression analysis on glutathione reductase data indicated that the optimal dietary AsA requirement of L. vannamei was estimated to be 306.39, 319.75 mg kg^?1 diet respectively.     

The effects of diet supplemented with Lactobacillus rhamnosus on tissue parameters of rainbow trout,Oncorhynchus mykiss (Walbaum)

This study was carried out to establish the effects of a 6 week treatment with the diet supplemented with L. rhamnosus in concentrations of 10⁷ CFU g^?1 (G1 group) and 10⁸ CFU g^?1 (G2 group) on the condition expressed by condition factors (Fulton's, Clark's and B), intestinal microbiology, haematological, histological and selected antioxidative parameters of rainbow trout. A significantly higher condition factors were found in G1 group indicating that higher concentration of probiotic (10⁸ CFU g^?1) did not result in the better condition. Cholesterol and urea levels were significantly higher in both G1 and G2 groups, albumin in G1 and creatinine in G2 group with respect to control. A significantly higher liver TBARS level was observed in G2 group. The feeding with supplemented probiont apparently changed the resident microbiota. Three weeks after withdrawal of the supplemented feed, the microflora mostly reverted to the control composition, although L. rhamnosus in faecal matter of fish remained inherent. The epithelial structure of the proximal and distal intestine revealed the increased absorptive area in both treated groups, as well as the increase in the mucin‐secreting goblet cells. The L. rhamnosus‐treated groups demonstrated the capacity for the augmentation of the innate host defence.     

Phenotypic indicators for cultured pearl size improvement in the black‐lipped pearl oyster (Pinctada margaritifera): towards selection for the recipient growth performance

The black‐lipped pearl oyster, Pinctada margaritifera, is the most important farmed species in French Polynesia and the basis of the most valuable export industry. Mass production of black pearls relies on a surgical operation requiring tissue from a donor pearl oyster to be grafted, together with a nucleus made of shell, into the gonad of a recipient oyster. Improving pearl size through family selection remains one of the main challenges for future aquaculture development. This study analyses the relative contribution of donor and recipient oysters to pearl size. To this end, hatchery‐produced donor oysters of two batches, large and small (based on shell height), were used to supply grafts for recipients, which were then monitored individually for their growth performance by recording shell height, width, and thickness, and total live weight (flesh + shells) every 6 months (four biometric measurement times) over 20 months of culture. Pearls issued from the two batches of donors showed no significant differences in nacre weight or thickness. In contrast, recipient oyster shell height and total weight were increasingly positively correlated with these pearl size parameters over the culture period, becoming significant at 8 months post‐grafting. Potential therefore exists to use shell height and oyster weight as phenotypic indicators for selective breeding of recipient oysters with high growth performance to increase pearl size in P. margaritifera.     

Effect of dietary protein content on growth rate,survival and body composition of juvenile cauque river prawn,Macrobrachium americanum (Bate 1868)

We determined the effect of four concentrations of dietary crude protein, 30.7, 37.2, 41.8 and 46.8% on growth rate, survival and body composition of the juvenile cauque river prawn (Macrobrachium americanum). The prawns were hatched in the laboratory from the spawn of one wild ovigerous female. Prawns consuming 37.2% crude protein reached a final weight of 0.58 g (feed conversion ratio of 2.15), which was significantly better than the other treatments. Survival was 100% in all treatments. Protein content in the diets had no significant effect on whole body proximate composition and amino acid profile. Juveniles consuming the 37.2% crude protein diet grew faster than those fed the other diets. Specific growth rate was adjusted to the two‐slope broken‐line regression analysis model to estimate the optimal protein requirement. In conclusion, these results demonstrated that the 37.2% protein level diet is optimal for juvenile cauque river prawn M. americanum in the experimental conditions of this study.     

Induction of meiotic gynogenesis in bagrid catfish (Pseudobagrus ussuriensis) with homologous sperm and its confirmation for female homogamety

The bagrid catfish, Pseudobagrus ussuriensis, exhibits significant sexual dimorphism in growth rate and body size with males growing faster than females. Therefore, an all‐male culture can dramatically increase the output and profitability of fishery products. According to the monosex breeding route, super‐male individuals’ acquirement by XY male sex reversal and artificial gynogenesis is the key step. An effective protocol to induce meiotic gynogenesis using homologous sperms has been developed in this study. The most effective UV irradiation for sperm genetic inactivation was found to be at a distance of 20 cm with 66 μW/cm² light intensity for 25 min. Optimal treatment for cold shock was 5 min post‐fertilization at 0‐4°C for 30 min, which gave the best survival rate of 13.65 ± 2.87%. The sex ratio in the meiotic gynogens showed a significant female‐biased deviation (p < .05); thirty meiogynogens and their parents were further analysed using a male‐specific AFLP marker, of which only the male parent produced a male‐specific DNA band of 412 bp. These results indicated the female homogametic XX/XY sex determination system in P. ussuriensis. The optimization of a protocol for the successful induction of meiogynogenesis in the bagrid catfish lays the basis for all‐male production and is useful in ascertaining the genetic sex determination system in this promising aquaculture species.