The cellular community in the intestine of the shrimp Penaeus penicillatus and its culture environments

In the present study, bacterial community diversities were investigated in the shrimp intestines, outside seawater and culture-associated environments using clone library analysis. Comparative analyses revealed that the bacterial community in the intestinal contents was most similar to that of the sediment. Bacteria in the phyla Proteobacteria and Firmicutes constituted the dominant components of the intestinal bacteria in the shrimp. The potential pathogens and probiotics were dominant members of the intestine in healthy or diseased shrimp. Intestinal bacterial communities in the healthy shrimp were composed of Bacillales, Rhodobacterales, Vibrionales, and Alteromonadales, whereas fewer bacterial groups were observed in the intestines of diseased shrimp, which were dominated by Vibrionales and Bacillales. Phylogenetic analysis revealed that nearly half of the sequences in the intestines of diseased shrimp were closely related to the potential pathogen Photobacterium damselae and Vibrio harveyi. Obvious differences in microbial diversity were also observed between outside seawater and culture-associated environments. Eukaryotes in the outside seawater were dominated by Skeletonema, Cyclotella, Thalassiosira, Guinardia, and Chaetoceros. The pond water showed less diversity in the eukaryotic community and was dominated by Skeletonema, Thalassiosira, Chaetoceros, and Oithona. The implications of the present study may be significant in the management of shrimp cultivation, as well as the detection of potential pathogens and the application of probiotics.     

Origin of the diclidophorid monogenean Neoheterobothrium hirame Ogawa, 1999, the causative agent of anemia in olive flounder Paralichthys olivaceus

In the mid-1990s, Neoheterobothrium hirame suddenly appeared as a new species in olive flounder Paralichthys olivaceus in Japanese coastal waters. Anemia caused by the parasite has prevailed in wild and cultured populations of olive flounder since that time. In this study, to clarify the origin of N. hirame, two Neoheterobothrium species, namely unidentified Neoheterobothrium species (tentatively abbreviated as Neoheterobothrium sp. PL) and N. affine, were collected from Paralichthys lethostigma and Paralichthys dentatus, respectively, off the east coast of North America and compared with N. hirame collected in Japan. No substantial differences were detected in the morphology and DNA sequences of ITS1-5.8S rRNA-ITS2 and mitochondrial cytochrome oxidase subunit I (mt COI) regions between N. hirame and Neoheterobothrium sp. PL. On the other hand, the congeneric N. affine was clearly distinguished from both N. hirame and Neoheterobothrium sp. PL in its longer isthmus and the DNA sequences in ITS1 and mt COI. The absence of differences between N. hirame and Neoheterobothrium sp. PL and the clear difference between both of these and N. affine indicate that N. hirame is conspecific with Neoheterobothrium sp. PL infecting P. lethostigma and that N. hirame was recently introduced from North America to Japan.     

Escapement from the main body of the bottom trawl used for the Mediterranean international trawl survey (MEDITS)

Escapement through the body or under the footrope of the sampling trawl used for the Mediterranean international trawl survey programme (MEDITS) was assessed i) by means of visual observations performed with a towed operating vehicle equipped with a high-sensitivity video camera, and ii) by attaching small bags to the outside of the trawl body and under the footrope. Due to the small size of most of the individuals present in the study area and to the towing speed, fish behaviour in front of the trawl could not be frequently observed. In contrast, their behaviour was easily observed inside the net, especially upstream and inside the codend. Some species, such as horse mackerel (Trachurus sp.), showed greater swimming endurance compared with others such as annular seabream (Diplodus annularis). In three fishing trips conducted in different seasons in 1996 and 1997 to obtain data on several species in different areas of the Adriatic, six small net bags were mounted outside the body of the MEDITS trawl to quantify escapement through the trawl belly and under the footrope. Significant escapement of many species occurred in the near-bottom part of the side panels. Norway lobster (Nephrops norvegicus) had the highest rate (64 %), mainly represented by small-size individuals. The escapement of greater forkbeard (Phycis blennoides) was also high (53 %). For hake (Merluccius merluccius), common pandora (Pagellus erythrinus) and poor cod (Trisopterus minutus capelanus), rates ranged from 10 to 16 %. The escapement of red mullet (Mullus barbatus) was very low despite their great abundance. Blue whiting (Micromesistius poutassou) escaped mainly through the mid-height part of the side panel. This great variability of escapement values could affect the proportion rates of the species sampled during a standard survey. Different escape behaviours among size classes were observed for M. poutassou, T. minutus capelanus and P. erythrinus, with smaller individuals tending to escape towards the bottom and larger ones towards the higher parts of the trawl body. In the species where the escape rate was size-dependent, therefore, the size-frequency distributions obtained from the codend catch may not reflect actual size-frequency distributions.     

Effect of Rhodomonas salina addition to a standard hatchery diet during the early ontogeny of the scallop Pecten maximus

The main objective of this study was to identify algal diets that maximize the survival and growth and alter the biochemical content of Pecten maximus larvae with the aim of improving metamorphosis. We also evaluated the potential of the cryptophyceae Rhodomonas salina as a food source for these larvae. Two flagellates, Isochrysis aff. galbana (T) and Pavlova lutheri (P), and two diatoms, Chaetoceros gracilis (C) or Skeletonema costatum (S), were tested as two ternary diets, namely PTC and PTS. PTC and PTS were compared with diets that also included R. salina (R). The addition of R. salina and the replacement of C. gracilis by S. costatum in the traditional hatchery diet seem to be two interesting alternatives for increasing the productivity of larval scallop culture and improving the metamorphosis rate. With these two diets, larval growth increased and metamorphosis was observed to occur more rapidly. Moreover, our results showed that the addition of R. salina significantly improved the overall condition of the larvae by promoting an increase in organic matter and total lipids. This accumulation of lipids during ontogeny seems to promote larvae to grow and to complete metamorphosis more rapidly than with the other diets. The level of polyunsaturated fatty acids in the algae could also at least partially explain the results obtained, since the PUFA level of C. gracilis was about half those of S. costatum and R. salina.     

A comparison of the gametogenic cycle between the Pacific oyster Crassostrea gigas and the Suminoe oyster Crassostrea rivularis in Washington State

The gamotogenic cycle of the Suminoe oyster, Crassostrea rivularis, was compared with that of two groups of Pacific oysters (Crassostrea gigas) on an intertidal oyster bed in Oakland Bay, Washington. The Pacific oysters exhibited an earlier and more extensive development of gonad relative to C. rivularis. Resulting carbohydrate depletion was much greater in C. gigas relative to C. rivularis. Both groups of Pacific oysters spawned completely in mid-August. C. rivularis did not spawn but, rather, initiated phagocytosis of gametes by leucocytes. The implications of the results on the development of a marketable “summer” oyster are discussed.     

Dietary supplementation with Bacillus can improve the growth and survival of the kuruma shrimp Marsupenaeus japonicus in high-temperature environments

This study examined the effects of a commercial Bacillus complex used as probiotics on the performance of Marsupenaeus japonicus under high water temperature after 60 days of rearing. Growth and survival rate of M. japonicus were significantly increased in the probiotics-treated group (P < 0.05). Vibrio count and percentage of Vibrio in the intestinal tracts of M. japonicus reared with Bacillus were significantly lower than in other treatments (P < 0.05). The expressions of immune-related gene including prophenoloxidase, lysozyme, cytosolic manganese superoxide dismutase and hemocyanin subunit L in M. japonicus were significantly increased in the Bacillus-treated group (P < 0.05). The results indicated that Bacillus can be used as a diet supplement to minimize the damages caused by free radicals generated from insufficient oxygen metabolism due to the high-temperature stress, and to enhance immunity and activate the immune response levels. Our findings can improve the growth and survival rates of M. japonicus during the high-temperature farming period.     

Feeding experiments on the larvae of the fiddler crab Uca pugilator (Brachyura,Ocypodidae), reared in the laboratory

Larvae of Uca pugilator (Bosc) were reared in the laboratory from hatching to the megalopa stage on three different diets: (1) newly hatched Artemia salina nauplii (diet A), (2) the rotifer Brachionus plicatilis (O.F. Müller) and a ciliate Euplotes sp. (diet RC), and (3) a combination of the above two diets (diet ARC). The survival rate of zoeae fed diet A (90.0%) and diet ARC (93.8%) was much higher than that of the larvae fed diet RC (22.5%). The duration of the zoeal stages was significantly shorter for the larvae fed diet ARC than for those fed diets A and RC. The survival rate of megalopa larvae (reared on diets A and ARC in the zoeal stages) was high (above 90%) for megalopa fed Artemia nauplii only, as well as for those fed a combination of Artemia nauplii and shrimp. No significant differences in duration of the megalopa stage were found between the latter diet groups.     

Relationship between the water molecules in fish-meat gel and the gel structure

To elucidate the structure of the network of the water retention mechanism of fish-meat gel, the microscopic relationship between the water content and viscoelasticity was investigated, and the specific surface area (S) where the water molecules are adsorbed was also measured. The gel structures of fish-meat, actomyosin (AM), and myosin (M) were investigated. In the fish-meat gel, as the water content increased, the number of network chains (??) decreased and the molecular weight between the cross-linking points (M _e) increased. At 10?% salinity and approximately 79?% moisture, ?? decreased to a minimum and M _e began to increase remarkably. In addition, when the moisture increased to 78?%, S increased about 1.5 times and the elasticity decreased significantly, indicating that this level of water content is the limit for sufficiently forming a gel structure in fish-meat gel. In the AM and M gels, M _e and S were similar to those observed in the fish-meat gels. When the moisture content increased, they also increased, regardless of the salt concentration. This result suggests that the size of the network was enlarged or that the network chains were cleaved.     

Preliminary studies on the growth and food consumption of the juvenile loggerhead turtle (Caretta caretta L.) in captivity

The effects of temperature and food ration on the growth of juvenile loggerhead turtle, Caretta caretta L. were examined in relation to body weight under controlled conditions in the laboratory. With increasing food ration there was increasing growth rate. The best daily growth rate at 20°C water temperature was 1.87%. Basal metabolism was reached at a daily feed rate of 1.01% body weight at 20.5°C derived from a formula for growth, namely g = Af ? B where g = 0.62f ? 0.63.     

Dealing with the presence of the ciliate Euplotes sp. in cultures of the copepod Acartia tonsa

Ciliates in live feed cultures can be a pest that lower production yields. This could dramatically affect the management and success of copepod cultures. In this study, we investigated the effect of the ciliate Euplotes sp. on egg production, specific egg production and egg hatching success of Acartia tonsa fed with Rhodomonas salina. We found that at a concentration of 2 cells ml^?1, Euplotes sp. had no effect on the production and hatching success of eggs but increased/decreased the mortality/quality of non-subitaneous eggs. Euplotes sp. had a good fatty acid profile containing high proportion of unsaturated fatty acids, but the amount of fatty acids compared to their carbon weight was very low and, therefore, it was considered as a poor food source. We propose a short-term interruption of food supply to copepod cultures in order to mitigate bloom formation of ciliates. This will force copepods to feed on them actively as prey and therefore decrease their long-term negative effects.     

The effects of food and the sum of effective temperature on the embryonic development of the seahorse,Hippocampus kuda Bleeker

The effect of different food (live Acetes spp., live Mysis spp., frozen Mysis spp., and mixed food of 50% live Acetes spp. and 50% frozen Mysis spp.) on gonad development of seahorses, Hippocampus kuda Bleeker, was evaluated in this experiment. The developmental durations of testes and ovary of seahorses were significantly different among the four experimental treatments. The live Acetes spp. treatment presented the shortest developmental durations to stage V, which were 87.6 ± 3.84 days and 89.2 ± 3.71 days, respectively, for ovary and testes. The frozen Mysis spp. treatment had the longest developmental durations (F_3,15 = 13.284, P < 0.05). The relationship between developmental duration of the ovary and gonad developmental stages could be formulated: Y₁ = 12.04x + 24.36 (r² = 0.9722, n = 16, P < 0.05). The gonadosomatic index (GSI) of parent seahorses among the four treatments differed significantly (F_3,15 = 18.364, P < 0.05). The standard GSI of seahorses fed live Acetes spp. was 15.64 ± 1.65%, which was the highest. Feeding live food had a significant effect on the fecundity and spawning of seahorses. The fecundity and spawning number of the live Acetes spp. treatment were 598 ± 45.49 and 552 ± 49.19 individuals, respectively, which were dramatically higher than those of frozen treatment (F_3,19 = 34.152, P < 0.05). Live food also had a similar effect on the fertilization and hatching rate during the embryonic development of seahorses (F_3,19 = 11.386, P < 0.05). Food treatment could also induce an indirect effect on survival rate of juveniles through gonadal and embryonic development of the parents (F_3,15 = 14.519, P < 0.05). In this experiment, the mortality within parent seahorses in the frozen Mysis spp. treatment was the highest (15.1 ± 6.55%), and the survival of juveniles was the highest in the live Acetes spp. treatment (90.4 ± 2.26% at 10 days). In conclusion, feeding live Acetes spp. significantly benefited the gonadal and embryonic development of H. kuda.The effect of temperature (22 °C, 24 °C, 26 °C, 28 °C, 30 °C and 32 °C) on the hatching time of H. kuda was also studied. We demonstrated that the higher the temperature, the shorter the hatching time, as well as the higher the hatching speed. The relationship between hatching time and temperature could be expressed: T = − 39.337 t + 677.75 (r² = 0.9755, n = 30, P < 0.05). In this finding, we provided the sum of effective temperature (SET) and threshold temperature of embryonic development of H. kuda (14066.9 °C h^− 1 and 13.7 °C, respectively). This new information on the effect of feeding type and culture temperature is beneficial for the commercial rearing and breeding industry of this species.     

Effect of antiprotozoal drugs on the proliferation of the bivalve parasite Perkinsus olseni

The protozoan parasite Perkinsus olseni causes severe losses among Ruditapes decussatus clams. The development of an in vitro culture of this parasite together with the use of a proliferation assay has provided the opportunity to screen for drug sensitivity of this parasite. Xenobiotics known for their antimalarial and antiprotozoal properties were tested against P. olseni. Only four of these drugs, namely cycloheximide, pyrimethamine, deferoxamine (DFO) and 2,2-bipyridyl (BIP), showed in vitro inhibitory effect on the parasite proliferation. Two in vivo experiments were designed to determine the effect of iron chelators on reducing P. olseni infection in clams. For this purpose, naturally infected clams from the Ria Formosa, Portugal, were exposed to DFO and BIP at various concentrations. In the first experiment, hemolymph samples were taken from each clam before and after treatment to determine the infection intensity and in the second experiment, clams were randomly distributed in groups of five and the parasite burden in treated and untreated groups was determined at the end of the experiment on the whole clam wet tissues. Only DFO was found to be effective in reducing in vivo P. olseni infections. In addition, acute toxicity of DFO and BIP has been determined and no mortality of Perkinsus-free clams was observed.     

Diversity of denitrifying bacteria in the greenwater system of coastal aquaculture

Greenwater technology is based on integrating finfishes inside pens for zero water exchange system of shrimp aquaculture. Nitrogen transformation could be improved by rearing euryhaline finfishes like grey mullet, milkfish which have a broad diet and tolerate poor water quality. The abundance of four denitrifying functional genes coding for nitrate reductase (narG and napA), nitrite reductase (nirS), nitric oxide reductase (qnorB) and nitrous oxide reductase (nosZ) has been examined in the greenwater system through a metagenomic approach. Phylogeny revealed homology of narG clones with uncultured environmental clones, whereas napA clone sequences were found to have homology with cultured (Stappia aggregata) and uncultured microorganisms. The nirS clones show uniqueness with Marinobacter hydrocarbonoclasticus, Aromatoleum aromaticum, and Ruegeria pomeroyi. The qnorB gene has been reported for the first time from culture systems along the Indian coast and clone exhibited 84–87 % identity with different uncultured bacteria. The nosZ clones are closely affiliated with S. aggregate and alpha bacterium. This study revealed denitrifying diversity from a greenwater system which could eventually be used in planning the future strategy for comprehending nitrogen fluxes, greenhouse gases and their mitigation in coastal aquaculture systems.     

The effects of green and blue-green algal diets on the reproductive rate of the rotifer Brachionus plicatilis

The reproductive rate of the rotifer Brachionus plicatilis cultured on a variety of diets was investigated. Unialgal and mixed diets of the green alga Chlorella and the blue-green alga Schizothriz were compared. Rotifer reproductive rate was found to be an average of 2.7 times higher on a mixed diet of Chlorella and Schizothrix than on either Chlorella or Schizothrix as a unialgal diet. Enhancement of rotifer reproductive rate was not observed on a mixed diet of Chlorella and Dunaliella compared with a unialgal diet of Chlorella. Ingestion of Schizothrix was shown not to be required for enhancement of rotifer reproduction. It was further shown that the enhancing factor is a heat labile substance.     

尼罗罗非鱼雌雄鱼肌肉组织差异表达基因的筛选

应用引物退火控制技术(ACP)筛选尼罗罗非鱼雌雄鱼肌肉组织差异表达基因,寻找与雌雄鱼肌肉生长发育相关的候选基因。本实验从同等条件下养殖的尼罗罗非鱼群体中随机选取雌、雄鱼各5尾组成RNA池,采用引物退火控制技术,分析了两组个体肌肉组织差异表达基因。利用20对随机引物差异显示扩增,共获得8条ESTs,其中5个已知的ESTs分别为转录变体3(LOC100691543)、60S核糖体蛋白(RL3)、小白蛋白β样蛋白、肌型肌酸激酶M2-CK和转录因子Sox4,其余3个为未知的ESTs。实时定量PCR分析各差异表达基因在尼罗罗非鱼雌雄肌肉组织中的表达发现,8个差异表达基因中转录变体3与ACP6-Y在尼罗罗非鱼雄鱼肌肉组织中的表达均极显著高于雌鱼(P0.01),ACP3-X、60S核糖体蛋白(RL3)、小白蛋白β样蛋白、ACP15-X、肌型肌酸激酶M2-CK与转录因子Sox4在尼罗罗非鱼雌鱼肌肉组织中的表达均极显著高于雄鱼(P0.01)。结果表明,应用引物退火控制技术筛选获得了8个可能参与了雌雄鱼肌肉生长发育调控的ESTs,为进一步筛选雌雄鱼肌肉生长发育相关候选基因奠定了基础。     

The effect of nitrate supplementation on the biochemical composition of benthic diatoms and the growth and survival of post-larval abalone

Elevating the nitrogen concentration in microalgal growth media can elevate the protein content of the algae and consequently increase their dietary value to molluscs. This study examined the protein content, and dietary value to abalone post-larvae, of seven benthic diatoms acclimated to high-(“H” = 49.4 mg NO₃-N l^− 1) and standard-nitrate (“S” = 16.5 mg NO₃-N l^− 1) Walne medium. In Experiment 1, the level of nitrate in the culture medium did not significantly affect the protein content of Cocconeis sp., Cylindrotheca closterium or Nitzschia ovalis, and variation in the growth of post-larval Haliotis iris was not associated with nitrate level or the protein content of the diatoms. When the same six diatom diets were added daily as cell suspensions, growth and survival were much lower than for the equivalent strains fed as established films (P < 0.0001). In Experiment 2a, the protein content of Navicula incerta was elevated by nitrate supplementation, and the high nitrate strain retained a higher protein content (P = 0.005) even after 24 days growing in unenriched, flowing seawater. Haliotis rufescens post-larvae grew significantly faster when fed N. incerta H than when fed N. incerta S (P = 0.006) and survival was also higher with this diet (67 versus 54%). In Experiment 2b, there was no significant effect of nitrate level on protein content of four diatoms or the performance of post-larval H. rufescens. There was, however, a strong positive correlation (r = 0.95, P < 0.0001) between post-larval survival and the lipid content of the diatom diets. Post-larval growth showed positive, but marginally non-significant, relationships with diatom protein and carbohydrate, but no relationship with lipid content. Overall, only one of seven benthic diatom species acclimated to high-nitrate growth medium displayed significantly elevated protein content. Diatom protein content explained relatively little of the variation in food value of diatom strains for post-larval abalone but lipid content correlated strongly with post-larval survival.     

The culture of the red algal genus Eucheuma in the Philippines

The author discusses the recent development and present status of a Eucheuma seaweed farming industry in The Philippines. He describes Eucheuma cultivation methods, assesses farm productivity, and considers the economics of Eucheuma farming by Filipino families. The author concludes with a discussion of current problems and an evaluation of future prospects for the industry.