Cadmium toxicity to embryonic-larval development and survival in red sea bream Pagrus major

At 18 degrees C and 33 psu, 24 and 48 h LC50 values of cadmium (Cd) for red sea bream Pagrus major embryos were 9.8 and 6.6 mg l(-1), respectively, while 24,48, 72, and 96 h LC50 values for larvae were 18.9,16.2, 8.0, and 5.6 mg l(-1), respectively, indicating that embryos were more sensitive to Cd toxicity than larvae. Cd concentrations at >= 0.8 mg l(-1) led to low hatchability (0-90% in >= 0.8 mg l(-1) solutions vs. 97-100% in lower ones), delay in time to hatch, high mortality (38-100% vs. 1-10%), morphological abnormality (42-100% vs. 1-10%), reduced length (3.55-3.60 vs. 3.71-3.72 mm) in the embryos and larvae. They were Cd concentration dependent and potential biological significant endpoints for assessing the risk of Cd to aquatic organisms. Heart beat and yolk absorption of the larvae were significantly inhibited at some high concentrations but they were not as sensitive as other endpoints to Cd exposure. (C) 2008 Elsevier Inc. All rights reserved.

Cloning, characterization and expression of ferritin subunit from clam Meretrix meretrix in different larval stages

Shell formation is one of the important events during larval development and metamorphosis in bivalves. However, the molecular mechanisms and environmental cues regulating shell initiation and growth are unclear. Here, we report that ferritin, a principal protein for biological iron storage and metabolism, might play a role in larval shell development of the bivalve mollusk Meretrix meretrix. A full-length ferritin subunit cDNA, named as MmeFer, was cloned and characterized. The MmeFer mRNA expression in different developmental stages, from trochophore to post larvae, was analyzed by real-time reverse transcription polymerase chain reaction (RT-PCR). MmeFer mRNA expression in larvae of later developmental stages increased at least 8-fold following trochophores. Moreover, the temporal and spatial expressions of MmeFer mRNA were examined by whole mount in situ hybridization. In the trochophore stage, MmeFer was detectable where it was supposed to be for shell initiation. In the later developmental stages, MmeFer was found near digestive glands and mantle that secret larval shell. MmeFer expression was also detected in larvae cultured in artificial seawater with different iron concentrations ranging from 0 to 100 mu M. These results suggest that ferritin may play a role in the shell formation of mollusks. (C) 2009 Elsevier Inc. All rights reserved.

A Dicer-1 gene from white shrimp Litopenaeus vannamei: Expression pattern in the processes of immune response and larval development

Dicer is a member of the RNAase III family which catalyzes the cleavage of double-stranded RNA to small interfering RNAs and micro RNAs, and then directs sequence-specific gene silencing. In this paper, the full-length cDNA of Dicer-1 was cloned from white shrimp Litopenaeus vannamei (designated as LvDcr1). It was of 7636 bp, including a poly A tail, a 5' UTR of 136 bp, a 3' UTR of 78 bp, and an open reading frame (ORF) of 7422 bp encoding a putative protein of 2473 amino acids. The predicted amino acid sequence comprised all recognized functional domains found in other Dicer-1 homologues and showed the highest (97.7%) similarity to the Dicer-1 from tiger shrimp Penaeus mondon. Quantitative real-time PCR was employed to investigate the tissue distribution of LvDcr1 mRNA, and its expression in shrimps under virus challenge and larvae at different developmental stages. The LvDcr1 mRNA could be detected in all examined tissues with the highest expression level in hemocyte, and was up-regulated in hemocytes and gills after virus injection. These results indicated that LvDcr1 was involved in antiviral defense in adult shrimp. During the developmental stages from fertilized egg to postlarva VII, LvDcr1 was constitutively expressed at all examined development stages, but the expression level varied significantly. The highest expression level was observed in fertilized eggs and followed a decrease from fertilized egg to nauplius I stage. Then, the higher levels of expression were detected at nauplius V and postlarva stages. LvDcr1 expression regularly increased at the upper phase of nauplius, zoea and mysis stages than their prophase. The different expression of LvDcr1 in the larval stages could provide clues for understanding the early innate immunity in the process of shrimp larval development. (C) 2010 Elsevier Ltd. All rights reserved.

Effects of various algal diets and starvation on larval growth and survival of Meretrix meretrix

Effects of food availability on larval growth and survival of Meretrix meretrix were studied in two experiments by feeding the larvae with different algae diets and by starving the larvae for different periods of time. Newly hatched larvae of M meretrix were fed with five different marine microalgae species, singly and in various mixtures. Best growth was with Isochrysis galbana as a single species diet. Nutritional value of the other single species diets was in the order of Dunaliella sp.> Phaeodactylum tricornutum > Platymonas subcordiformis > Pavlova viridis. Of the mixtures tested, 50% I. galbana/50% Dunaliella sp., 50% I. galbana/50% P tricornutum, and 50% 1 galbana/50% P subcordiformis, supported growth and metamorphosis equivalent to those of the I. galbana control. At 25 degrees C, larvae of M meretrix were deprived of food for various days to study the growth compensation from the outset of development. The results showed that M meretrix larvae could survive long feeding delays, and even reach metamorphosis without food added, although starvation had significant effects on growth. These results suggested that M meretrix larvae had the capacity to survive 'starvation' using alternative sources of energy. It also showed that growth, survival and metamorphosis of M meretrix were affected by many factors besides food quality and quantity. (c) 2005 Elsevier B.V. All rights reserved.

Pharmacological and immunocytochemical investigation of the role of catecholamines on larval metamorphosis by beta-adrenergic-like receptor in the bivalve Meretrix meretrix

Catecholamines regulate several physiological processes in mollusks. Many pharmacological experiments have been conducted to determine the effects of adrenergic agonist and antagonist of catecholamine receptors on Meretrix meretrix metamorphosis. Results showed that adrenaline (AD) and noradrenaline (NA) had substantial effects (p < 0.05) on larval metamorphosis at concentrations ranging from 10 mu M to 100 mu M. 10 mu M beta-adrenergic receptor (AR) agonist isoproterenol showed the same inducement effect as that of NA and AD on metamorphosis, whereas the alpha-AR agonist phenylephrine had no significant effect at concentrations between 0.1 mu M and 100 mu M concentrations (p > 0.05). Furthermore, I mu M beta-AR antagonist propanolol, but not alpha-AR antagonist prazosin, depressed the larval metamorphosis induced by NA or AD. By immunocytochemistry, two cell bodies of beta-adrenergic-like receptor, C/A1, C/A2, were observed in the cerebral/apical ganglion of competent larvae. In addition, there were other immunoreactive dots near C/A1 and C/A2. The results of pharmacology and immunocytochemistry suggests that beta-adrenergic-like receptor located in the larval CNS, might play a considerable role in the larval metamorphosis of M meretrix by AD or NA. (c) 2006 Elsevier B.V. All rights reserved.

Production of a base population and its responses to F-1 selection in the bay scallop, Argopecten irradians irradians Lamarck (1819)

A base population of the bay scallop, Argopecten irradians irradians Lamarck, was produced by crossing two cultured bay scallop populations. After 1 year of rearing, the top 10% truncation selection of the top 10% (i=1.755) was carried out in the base population of about 1300 adults. A control parental group with a an identical number to the select parental group was randomly selected from the entire population before isolation of the select parental group. The result showed that, at the larval stage, the growth rate of larvae in the selected line was significantly higher than that of the control (P < 0.05), and that the genetic gain was 6.78%. Owing to the lower density of control at the spat stage, the mean shell length of the control line was larger than that of the select line at day 100. When the same density was adjusted between two lines in the grow-out stage (from day 100 to 160), the daily growth rate of the selected line was significantly higher than that of the control line (P < 0.05). Survival of the select line was significantly larger than that of the control line in the grow-out stage. In conclusion, the results obtained from this experiment indicate that selective breeding from a base population with a high genetic diversity established by mass spawning between different populations appears to be a promising method of genetic improvement in bay scallop, A. irradians irradians Lamarck.

Effects of starvation on larval growth, survival and metamorphosis of Ivory shell Babylonia formosae habei Altena et al., 1981 (Neogastropoda : Buccinidae)

The impact of starvation on larvae of Ivory shell Babylonia formosae habei was studied in a laboratory experiment. Newly hatched veligers showed considerable tolerance to starvation due to their endogenous yolk material, and time to the point-of-no-return (PNR; the threshold point during starvation after which larvae can longer metamorphose even if food is provided) was calculated to be 104.5 h. However, starvation still affected larval growth, survival, and metamorphosis. Mean shell length of larvae increased 49.77 mum day(-1) for nonstarved, but only 11.13 mum day (-1) for larvae starved for 108 h. After larvae began feeding, their growth rates rapidly recovered to the level of the nonstarved following short periods of starvation (less than 48 h), but were inhibited and unable to ever reach the level of the nonstarved when being starved beyond 48 h. Percent metamorphosis was 53.75% for the nonstarved, but all larvae died before 10 days for those starved for 108 h. Starvation not only affected larval time to reach metamorphosis, but also caused the delay in the time to metamorphosis. For the nonstarved, larvae took only 11.5 days to reach spontaneous metamorphosis, but they took 20 days to reach spontaneous metamorphosis when starved for 96 h, and this duration of delayed metamorphosis reached 8.5 days. Furthermore, the importance of yolk material for maintaining larval survival of B. formosae habei during starvation periods is also discussed. (C) 2004 Elsevier B.V. All rights reserved.

A new three-phase culture method for Manila clam, Ruditapes philippinarum, farming in northern China

Studies on reproduction, hatchery management, and culture of Manila clams Ruditapes philippinarum were carried out in an attempt to optimize their culture conditions and techniques. Results from these studies led to the development of a three-phase culture method for Manila clam farming in northern China. The key components of the new method were: 1) early spawning and over-wintering indoors (greenhouse); 2) optimized larval culture conditions and techniques; 3) juvenile rearing in shallow, fertilized nursery ponds; 4) optimized stocking size and density and substrate for mudflat grow out. Broodstock were maturated indoors for a month from early April to early May. Primarily because of higher water temperatures in the greenhouse the clams spawned more than one month earlier than in the natural environment. From May to July, juveniles were reared for 1-2 months indoors to a size of 2.0-3.0 mm in shell length before being moved to outdoor, pre-disinfected, nursery ponds. Juveniles were then reared in the nursery ponds for one month to about 1.0 cm before being transferred to the mudflat for grow out. Juvenile clams in nursery ponds grew considerably faster than in the natural environment probably because of higher temperatures and more abundant natural food. During grow out, the clams were reared for 4-7 months until they reached a market size (3.0-3.3 cm). Juveniles produced after August were over-wintered in the greenhouse in which the water temperature was about 3 degrees C higher than that of the outdoor environment. Juveniles grew at an average rate of > 20 mu m day(-1), while in the natural environment no growth was observed during winter because of low temperatures. Juveniles in the greenhouse grew to 2-3 mm by the following March before being moved into outdoor nursery ponds. The three-phase culture method not only shortened the production period from spawn to market size from 24-36 months to about 10-14 months, but also prolonged the spawning season from 2 to 7 months, resulting in increased production of seed and market-size clams. Compared with the traditional method, the new method could increase the yield of market-size clams by 10-11 times, and increase the profit per ha mudflat by as much as 124 times and the profit per kg market-size clams produced by 13 times. (c) 2006 Elsevier B.V. All rights reserved.