Proteomics analysis of mitochondrial extract from liver of female zebrafish undergoing starvation and refeeding

Many fish species can withstand long period of food deprivation and consequently compensate for any weight loss by undergoing rapid growth during resumption of feeding. There is an interest in taking advantage of compensatory growth to reduce feed cost. In this present study, we attempted to compare the changes in hepatic mitochondrial proteome of zebrafish undergoing starvation and refeeding. Two‐dimensional gel separation and image analysis revealed a total of 65 spots that showed changes in expression after 15 days of starvation followed by 7 days of refeeding. A total of 35 proteins were selected for mass spectrometry analysis, resulting in the positive identification of 18 proteins. Identified proteins indicated that starvation resulted in reduction in glycolysis and increase in gluconeogenesis, while refeeding caused these activities to return to normal levels. Expression pattern of several proteins related to fatty acid and amino acid metabolism also suggested the utilization of non‐carbohydrate resources for energy during starving conditions. Proteins with chaperoning and antioxidative roles such as glucose‐regulated protein, paraxonase and heat‐shock protein were also upregulated in starved conditions.     

Interactive effect of two methyl‐group sources on the growth and phospholipids’ content of kuruma shrimp Marsupenaeus japonicus Bate post‐larvae

Six micro‐bound diets were formulated to contain three levels of choline chloride (CC) (0.0, 0.6 and 1.2 g kg⁻¹) and 2 levels of methionine (Met) (0 and 15 g kg⁻¹). Soybean protein isolates (SPI) were used as the main protein source for its limited Met content. A significant (P < 0.05) interaction was determined between CC and Met on the survival (S %), weight gain (WG%), specific growth rate (SGR % day⁻¹), feed efficiency ratio (FER), protein efficiency ratio (PER) and phosphatidylcholine (PC), phoshphatidylethanolamine (PE) and Met contents of the whole body of shrimp. The shrimp group did not receive either supplemental CC or Met showed lower (P < 0.05) values of the above‐mentioned parameters than other shrimp groups fed with 0.6 and 1.2 g kg⁻¹ supplemental CC with or without Met supplementation. The present study showed that supplementation of 1.2 g kg⁻¹ CC in the diets could compensate shrimp post‐larvae with the needed methyl group when received Met‐deficient diets. The study also assumed that the biosynthesis of PC in the shrimp’s body can be achieved by the methylation of PE through the S‐adenosylmethionine (SAM) pathway and/or through the cytosine di‐phosphoryl (CDP) choline pathway directly from dietary choline.     

Evaluation of two anaesthetic agents and the passive integrated transponder tagging system in Octopus vulgaris (Cuvier 1797)

Octopus vulgaris is a species of demand in the market with the potential to diversify European aquaculture. However, this species develops complex social interactions under culture conditions, which may have negative effects on its growth, survival and profitability. In order to understand its behaviour under such conditions, individual tagging systems allow a careful evaluation of biological parameters, such as growth and longevity. The present work describes a combined protocol (anaesthetic and tagging) for implanting subcutaneous passive integrated transponder tags (PIT). The effect of two anaesthetic agents in facilitating octopus handling is assessed: clove oil at 20–40–100 mg L^?1 and ethanol (96%) at 1–1.5–2%. The most suitable body location of PIT tags, its effect on growth and mortality, the addition of a stitch and the PIT retention rate after 2 months in floating cages were evaluated. It was concluded that immersion in seawater with 1.5% of ethanol at 22.3±0.5 °C is a suitable anaesthetic for this species. The results showed that the best‐selected PIT body location was the upper left arm III. No effect of the PIT tagging system was found on growth and survival when tagged and untagged octopuses were compared. It was observed after 2 months that the stitch did not induce an increase in the retention rate and 81–100% tag retention regardless of the dietary treatment.     

Effect of emerged shipment on the physiological condition of the adductor muscle in adult giant lion's paw scallop Nodipecten subnodosus (Sowerby 1835)

Currently, there is an increasing interest in shipping live adult scallops to markets and broodstock to hatcheries; nevertheless, information about the shipping effect on live scallops physiology is very scarce. In a previous study, a method in emersion was developed to ship scallop seed out of water, but it was not known whether this method is useful to transport adult organisms. As a consequence, the purpose of this study was to evaluate the effects of transportation by the emersion method on the physiological status of the adductor muscle of adult giant lion's paw scallop. Live specimens were packaged in a container and transported in emersion for 11 h. Six scallops were frozen in a farm and a similar number were frozen as soon as they arrived to the laboratory. Physiological indices were determined in each lot and the survival was estimated 24 h after re‐immersion. As a result of the transportation, a significant loss (P<0.05) of total carbohydrates, glycogen, adenosine 5′‐triphosphate and adenilated energetic charge, and a significant increase (P<0.05) in the free amino acids concentration were observed. Eighty‐eight per cent survival was achieved; therefore, we conclude that this method is appropriate for shipping live adult scallops of Nodipecten subnodosus.     

Dietary protein requirement of juvenile marbled spinefoot rabbitfish Siganus rivulatus

Rabbitfish are an Indo‐Pacific herbivorous marine fish that have good market demand and are suitable for aquaculture. The present work was performed to determine dietary protein inclusion necessary for optimal growth of juvenile rabbitfish Siganus rivulatus. Six diets with increasing levels of protein (10, 20, 30, 40, 50 and 60 g crude protein 100 g^?1 feed) and similar levels of gross energy (20 MJ kg^?1) were prepared and offered to S. rivulatus juveniles maintained in triplicate cages placed in two large water tanks for 49 days. Growth progressively improved with dietary protein for fish offered diets from 10% to 40% protein inclusion. Diets with greater protein levels did not improve fish growth beyond that observed in the 40% group. Daily feed intake, apparent protein utilization and feed conversion ratio decreased as dietary protein increased. Protein efficiency (PE) was greatest (1.47) in fish offered the 10% protein diet and least in fish offered the 60% protein diet (0.80). No differences in PE were observed among all other treatments (20–50%). Results of the present work suggest that minimum dietary requirement for suitable growth of S. rivulatus juveniles is 40% protein when digestible energy of the diet is 16–18 MJ kg^?1.     

Use of macroalgae supplemented with probiotics in the Haliotis rufescens (Swainson, 1822) culture in Northern Chile

In this study, we evaluate the use of macroalgae as vectors of probiotics bacteria into the digestive tract of abalone to improve their survival and growth. It is shown that when abalone Haliotis rufescens of different sizes were fed with a natural diet composed of fronds of the macroalga Macrocystis integrifolia supplemented with a mixture of Vibrio sp. C21‐UMA, Agarivorans albus F1‐UMA and Vibrio sp. F15‐UMA bacteria, there was a significant increase (P<0.05) in the average monthly growth rate and survival (%)in a period of 210 days, compared with the control without a probiotic supplement. The permanence of the probiotics in the digestive tract of the animals was monitored, and it was found that the number of culturable C21‐UMA and F1‐UMA bacteria decreased significantly (P<0.05) in recently weaned and adult abalone, and they almost disappeared completely on day 19 of the bioassay. However, the culturable Vibrio sp. F15‐UMA disappeared completely from the digestive tract on day 22 of the bioassay, and these were the bacteria that remained at the highest concentration compared with the other two bacterial strains over the experimental period. It is therefore shown that it is feasible to use a probiotic mixture to improve the profitability of the H. rufescens culture.     

A physiological and biochemical approach to selecting the ideal diet for Ostrea edulis (L.) broodstock conditioning (part A)

To select the best microalgae for Ostrea edulis conditioning, four single species diets were fed to batches of broodstock, which were then compared using physiological and biochemical approaches. Ingestion and absorption were first studied, showing that Chaetoceros gracilis and Skeletonema marinoï were efficiently ingested (4.9–5.3 mg g^?1 h^?1) and absorbed (1.9–2.5 mg g^?1 h^?1) while Tetraselmis suecica led to the lowest physiological responses (0.36 and 0.12 mg g^?1 h^?1 respectively). When O. edulis were fed any microalgae except T‐ISO with only 79 mg g^?1, gonads accumulated carbohydrates from 116 to 141 mg g^?1 and the extent of accumulation depended on the species supplied in the order T. suecica<C. gracilis<S. marinoï. When oysters were fed either of the diatoms (C. gracilis or S. marinoï), an efficient transfer of 20:5(n‐3) to all tissues, including the gonads, was recorded while no enrichment in 22:6(n‐3) occurred in all tissues (polar fraction) when oysters were fed T‐ISO. In contrast (22:5(n‐6)), a characteristic fatty acid of T‐ISO accumulated in all tissues, confirming its allocation despite particularly low ingestion (1.66 mg g^?1 h^?1) and absorption (0.32 mg g^?1 h^?1). When oysters were fed C. gracilis or S. marinoï an efficient transfer of cholesterol and campesterol from diatoms to the gonads was observed, while no sterol accumulation occurred in the gonad when fed T. suecica. Because of low ingestion, absorption and poor biochemical compounds' transfer, T. suecica is not recommended for O. edulis conditioning. T‐ISO also exhibited low physiological performances but due to a specific 22:6(n‐3) enrichment in the gonad neutral fraction (16.1%), its potential role in reproduction should be considered. Chaetoceros gracilis is highly recommended for O. edulis broodstock while a source of DHA other than that provided by T‐ISO should be found due to its poor absorption. Because a mixed diet has been shown to be more efficient for O. edulis broodstock conditioning, complementary trials dealing with the effects of other species rich in 22:6(n‐3) such as Rhodomonas salina or Pavlova lutheri should be performed.     

Karyotype analysis of interspecific hybrids between Haliotis rufescens and Haliotis discus HANNAI

Evidence of hybridization in Haliotis has been mainly supported by hatchery experiences and collection of wild hybrid abalones among several species from natural populations worldwide. However, despite the importance to understand the role of the hybridization process through Haliotidae evolution, and also its impact on the abalone aquaculture, genetic studies in hybrid abalones have been poorly developed. Herein, cytogenetic approach allows studying the genetic conformation in hybrid organisms at the chromosome level. This paper reports a quantitative karyotype analysis in Haliotis rufescens, Haliotis discus hannai and their interspecific hybrid. Thus, to characterize chromosome pairs and establish cytogenetic comparisons, chromosome banding with distamycin‐A/4,6‐diamidino‐2‐phenylindole fluorochromes and morphologic measurements were performed. The results showed that the hybrids are successfully viable and their karyotypes evidenced a conservative chromosome number of 2n=36. The karyo‐idiogram showed a high correspondence in chromosome pair morphology among the hybrids and their parental species, except for a single heteromorphic pair that corresponds to the chromosome 16 from H. rufescens andH. d. hannai respectively. The implications of the abalone hybrid viability derived from its chromosome composition are discussed.     

Development of digestive biochemistry in the initial stages of three cultured Atherinopsids

The early development of the digestive biochemistry of three cultured Atherinopsids (two Odontesthes species and Chirostoma estor) was investigated. Relatively high lipolytic and high alkaline proteolytic activities were found in all species. In addition, gene expression of lipase and trypsin during the early development of C. estor not only indicates an early onset of the lipase activity but also demonstrates a major importance of protein digestion in this species. However, despite all of them share similar habitats, differences in their digestive activities were evident. The rise in the activities of brush border membrane enzymes correlated with the decrease in cytosolic activities, used as indicator of maturation of the digestive tract, was detected in both species of Odontesthes at 9 weeks after hatching. In contrast, no similar signs were measured in C. estor, this being in agreement to the long weaning period (nearly 5 months) reported for this species.