Contribution of the symbiotic dinoflagellate Symbiodinium microadriaticum to the nutrition,growth and survival of larval and juvenile tridacnid clams

The role of the symbiotic alga Symbiodinium microadriaticum (= zooxanthellae) in growth, development, and survival of larval and juvenile tridacnid clams was investigated. The zooxanthellae were not present in eggs released from adult Tridacna gigas and Hippopus hippopus, but were ingested by veliger larvae after development from the trochophore stage. Growth of veligers of the clam H. hippopus fed freshly-isolated S. microadriaticum was significantly higher than the growth of starved veligers. In contrast, veligers fed cultured strains of S. microadriaticum and 1–5-day-old isolates of S. microadriaticum from H. hippopus had growth and survival rates not significantly different from starved veligers. These results suggest that translocated photosynthetically-fixed carbon from the ingested freshly-isolated algae was responsible for increased veliger growth and survival.The presence of zooxanthellae in the stomachs of veligers before metamorphosis facilitated establishment of a symbiosis with S. microadriaticum and a concurrent increase in growth after metamorphosis. The establishment of the symbiosis with S. microadriaticum entailed passage of the algae from the stomach, through the digestive gland, and into the developing haemal sinuses. The contribution of photosynthetically-fixed carbon from the symbiotic zooxanthellae could theoretically account for the respiratory demand of 6-week-old juveniles of H. hippopus, corroborating positive growth of unfed symbiotic juvenile clams maintained in filtered seawater.When offered a variety of free-living and symbiotic species of algae, juvenile H. hippopus established a symbiosis only with strains of S. microadriaticum. Other species of algae were apparently digested if small enough to be ingested. Less than 7% of the free-living phytoplankton species Isochrysis galbana and Platymonas subcordiformis that were ingested by symbiotic juvenile Tridacna gigas (3–12 months old) were passed intact through the digestive system after 3 days. In contrast, when freshly-isolated zooxanthellae from T. gigas were offered to symbiotic juvenile T. gigas, up to 76% of the zooxanthellae ingested passed through the clam's digestive system and were released in faecal pellets intact. The symbiotic dinoflagellate Amphidinium klebsii and the diatom Phaeodactylum tricornutum were not ingested by veliger or juvenile clams.     

Dietary habits of grazers influence their suitability as biological controls of fouling macroalgae in ex situ mariculture

The dietary habits of the sea urchin Salmacis sphaeroides and the gastropod Trochus maculatus in ex situ mariculture were examined. Fouling algal assemblages on terracotta tiles were exposed to the grazers and compared. Except for Day 0, the assemblages on Days 2, 13 and 27 differed significantly across treatments. S. sphaeroides rapidly consumed most algae, leaving an assemblage dominated by turf algae and bare tile. T. maculatus primarily fed on green filamentous algae, resulting in an even distribution of other algal functional groups, while control tiles were dominated by green filamentous algae. Using three representative fouling algae species (Bryopsis corymbosa, Hypnea spinella and Lobophora variegata), the consumption rates of both grazers and dietary preferences of S. sphaeroides were examined through a single‐diet and a choice experiment respectively. The single‐diet experiment revealed differential algal consumption rates for S. sphaeroides (H. spinella = B. corymbosa > L. variegata) and T. maculatus (B. corymbosa > H. spinella = L. variegata). The choice experiment showed that S. sphaeroides preferred H. spinella over the other two species. These experiments highlighted the importance of understanding the dietary habits of grazers as this has direct implications on their effectiveness as biological controls of fouling macroalgae in mariculture.     

The mass outdoor culture of macroscopic marine algae

Two species of macroscopic marine algae (Rhodophyceae) were grown and harvested continuously during 1974 in Fort Pierce, Florida. The seaweeds Gracilaria sp. and Hypnea musciformis were grown in both 600-l and 350-l tanks in the third stage of a nutrient removalaquaculture process that included microscopic marine algae and oysters in the first two stages. Mixtures of secondary-treated waste water and sea water constituted the initial nutrient source, with the seaweeds utilizing nutrients not assimilated by the microscopic algae and/or generated by the oysters. Standing crops of up to 7 900 g wet weight of Hypnea and 9 000 g Gracilaria were maintained. The Hypnea cultures could not be maintained in summer when tank temperatures reached 30°C, but Gracilaria grew during the entire year. On the basis of periodic harvesting, yields were consistently between 12 and 17 g dry weight per m² per day for both species. These yields are as high as those achieved by mass cultures of microscopic marine algae, or by fast growing commercial crops such as sugar and rice.     

Instantaneous effect of dibromomethane on metamorphosis of larvae of the sea urchins Strongylocentrotus nudus and Strongylocentrotus intermedius

A volatile chemical, dibromomethane (DBM), produced from red coralline algae is known as a chemical inducer of larval metamorphosis of the sea urchin Strongylocentrotus nudus. We performed experiments exposing DBM to the larvae of S. nudus and Strongylocentrotus intermedius through a hydrophobic membrane. Metamorphic rates resulting from different diluted DBM solutions and exposure times were ascertained. The highest metamorphic rate, more than 80% in both species, was found after 1 h exposure to 1/2 diluted DBM. With this dilution, more than 80% of S. nudus and S. intermedius larvae metamorphosed 1 h after start of the experiment after only 10 and 5 min exposure, respectively, which corresponded to the low concentrations of 52–61 ppm and 34–43 ppm DBM by GCMS analysis, respectively. These findings suggest that DBM has an instantaneous effect on high success of metamorphosis of larvae of S. nudus and S. intermedius.     

The effect of the addition of algae feeding stimulants to artificial diets for the sea urchin Tripneustes gratilla

A key concern when developing artificial aquaculture diets is their palatability. This study investigated whether the palatability of artificial diets could be improved by incorporating a small quantity of highly preferred natural foods as feeding stimulants. The preference of Tripneustes gratilla for marine plants with which it co-occurs naturally in New South Wales, Australia, was assessed in a laboratory choice feeding experiment. T. gratilla displayed significant preference for the brown alga Ecklonia radiata over five other algae and one seagrass when they were offered simultaneously. Total protein or energy in the plants was not found to account for this preference. However, T. gratilla showed no preference among E. radiata, Sargassum linearifolium and Ulva lactuca when they were dried. Three artificial diets were made by incorporating one of each of these dried seaweeds at 5% dry weight. Although not statistically significant, T. gratilla ate more than twice as much the artificial diet containing S. linearifolium compared to the control diet containing no algae in a choice feeding experiment. In a no-choice feeding experiment, T. gratilla consumed significantly more of the Ecklonia and the Sargassum diets than the control diet despite each of the diets containing approximately the same protein and energy levels. T. gratilla consumed 49 and 43% more protein, and 37 and 44% more energy, respectively, when fed with the Ecklonia and the Sargassum diets compared to the control diet. Juvenile T. gratilla grew significantly faster on a wet weight basis on the Sargassum diet than the control diet at an average increase of 2.4% per day. These results indicate that the small amounts of palatable seaweed added to the artificial diets act as feeding stimulants, increasing the acceptability of artificial sea urchin diets, boosting the protein and energy consumption, and significantly increasing the growth of T. gratilla. Moreover, the fast growth of T. gratilla achieved in this study also indicates that this species would be a good candidate for commercial aquaculture.     

Molecular identification of macroalgal fragments in gut contents of the sea urchin <Emphasis Type="Italic">Hemicentrotus pulcherrimus</Emphasis>

In order to improve the efficiency of stock enhancement programs for the sea urchin Hemicentrotus pulcherrimus, information on food algal species, which affect growth and gonad production greatly, is necessary. Since it is difficult to identify species from the macroalgal fragments within the gut contents of the sea urchin by microscopic observation, we tried to apply a DNA barcoding method for gut contents analysis. We used a partial rbcL gene sequence for taxonomic section and newly designed primer sets, respectively, for brown algae and for red algae. Direct sequencing of the PCR products was carried out. Species identification was based on the phylogenetic relationship. We could objectively identify four species and two taxonomic groups (genus or family) in brown algae, and two species and four taxonomic groups in red algae from the gut contents. Sargassum hemiphyllum was the most abundant brown alga in the gut contents but was not dominant in the study site. The result showed the importance of identification to the species level. In addition, red algal epiphytes were detected with brown algal fragments. The DNA barcoding method will enable the researchers to verify the important role of epiphytes as a potential food source.     

Impact of food type on growth,survival and reproduction of the cyclopoid copepod <Emphasis Type="Italic">Cyclopina kasignete</Emphasis> as a potential live food in aquaculture

This study compared the efficacy of different dietary algae on the growth and reproduction of the cyclopoid copepod Cyclopina kasignete, a potential live food species for fish larvae in aquaculture. The experimental diets for the copepod consisted of three monoalgal diets (Nannochloropsis oculata, Tisochrysis lutea and dry Melosira sp.) and two mixed algae diets (T. lutea?+?N. oculata, T. lutea?+?dry Melosira sp.). The experiment was carried out for 30 days, and the population growth, survival and reproductive performance (generation time, hatching rate, life spawning times, daily offspring production, eggs per sac, lifespan and sex ratio) were used to assess the responses of C. kasignete to different food types. Population growth, survival and reproductive capacities of C. kasignete were significantly affected by the mono and binary species of algal diets. The results showed that copepods exhibited superior growth, survival and productivity when fed on fresh T. lutea, dry Melosira sp. and a mixture of both species compared to other dietary treatments. Copepods produced comparable growth, survival and productivity when fed on diatoms (dry Melosira sp.) as a single or in combination with other algae. This study indicates that cyclopoid copepod C. kasignete grow fast and have the potential to serve as a live food for aquaculture. The algae T. lutea, dry Melosira sp. and their combination are appropriate food to sustain the growth and reproduction of this copepods in mass culture as a potential live food in fish hatchery.     

On the role of algae in larviculture of Macrobrachium rosenbergii

The possible roles of algae (i.e. “green water”) in facilitating Macrobrachium rosenbergii larviculture have been investigated.Larvae were incubated with ¹⁴C uniformly labeled Tetraselmis sp. and Phaeodactylum sp. No counts were incorporated by the larvae indicating that negligible quantities of algal cell material are ingested directly. In contrast, Artemia nauplii assimilated and digested appreciable quantities of algae under similar conditions.The algae rapidly removed ammonia that had been added to the larviculture medium even starting at levels toxic to the larvae (0.6 ppm). However, very high densities of Artemia (20/ml) caused ammonia accumulation even in the presence of the algae due to their own increased metabolic rate when fed upon algae and due to the reduced number of assimilating algae. Removal of the Artemia under these conditions was followed by rapid disappearance of the ammonia.The beneficial effects of Tilapia as a larviculture associate, reported elsewhere, was observed only in the presence of algae. Larval mortality actually increased in water that had supported the growth of Tilapia if algae were not present.It is concluded that algae facilitate the growth of Macrobrachium larvae only indirectly by removing toxic material, notably ammonia, from the medium. Under conditions where the Artemia are grown before contact with the Macrobrachium larvae and where other means of ammonia removal are employed, larval growth and development are not affected by the presence of algae and the latter can be dispensed with.     

Viability of a Freshwater Mussel (Elliptio complanata) as a Biomechanical Filter for Aquaculture Ponds I: Clearance Rate of Chlorophyll-α

The production and livelihood of aquaculture facilities depend on increased growth of species being harvested. Only 15%-25% of nitrogen and phosphorus are retained in farmed fish even when all feed is consumed. In both marine and freshwater aquaculture systems, bivalves have proven to be an inexpensive method for controlling algal growth through suspension feeding. With 25% of the native U.S. freshwater mussel species under federal protection, their use in aquaculture ponds could lead to further propagation and restoration of natural populations. Elliptio complanata is an important species in Delaware rivers and connected estuaries; a recent two-year study by the United States Geological Survey found that 98% of mussels found in the Delaware River were Elliptio complanata. The objective of this study was to calculate the clearance rate of E. complanata in a laboratory study. This would help us determine if mussels can serve as an addition to chemical and mechanical filtration regiments and thrive in an aquaculture setting. Using feeding chambers we investigated the effect of E. complanata on algae grown from an aquaculture pond and on a cultured alga over a four-hour period. Temperature, dissolved oxygen, and pH were monitored, and samples were than analyzed for total chlorophyll-α. The average of both the single cell using Chlorella vulgaris and mixed assemblage treatments chlorophyll-α clearance rate was 0.187 liters per hour per mussel. Prior to this experiment, these mussels were held for a period for two months in the holding system in the laboratory. The resulting low clearance rate as compared to the previous studies in the field is possibly due to the stress from holding these mussels. However, our study demonstrated that mussels have the potential to be used in an aquaculture setting and/or recreational ponds to help reduce algae populations that can occur as a result of eutrophication.     

Experiments of the rearing of larval turbot, Scophthalmus maximus L.

Experiments on the live food requirements of cultured turbot larvae are described. A mixture of six species of unicellular algae did not sustain early larvae, but the growth and survival of larvae fed with rotifers were considerably improved when the rotifers were feeding on Isochrysis galbana rather than on Dunaliella tertiolecta. Oyster (Crassostrea gigas) larvae offered alone or with rotifers, did not produce higher larval growth or survival than rotifers alone. Black tanks were found to be more suitable than white tanks for rearing larval turbot.     

Culture-independent analysis of the bacterial community during fermentation of pa-som,a traditional fermented fish product in Laos

Pa-som is a traditional salty fermented food made from varieties of freshwater fish with garlic and rice in Laos. A similar product, plaa-som, is made in Thailand. To investigate the fermentation process of pa-som, lactic acid production and pH were monitored during laboratory-scale production of pa-som. Furthermore, the variety of bacteria involved and their succession during fermentation were examined using culture-independent polymerase chain reaction (PCR) denaturing gel gradient electrophoresis targeting the 16S ribosomal RNA (rRNA) gene. d- and l-lactic acid production and a decrease in pH were observed in the samples after 1 day of fermentation. The lactic acid content and pH continued to increase and decrease, respectively, until 4 days of fermentation. Overall, six lactic acid bacteria and eight other indigenous bacteria species were detected during pa-som fermentation. Among the lactic acid bacteria detected, two Lactococcus and one Weissella species were detected in samples after 1–4 days of fermentation, suggesting that these three species play major roles in pa-som fermentation from the initial phase. This is the first report to investigate the fermentation process in pa-som at the microbial level. This research approach should be applicable to a wide variety of pa-som and related fermented fishery products in Laos to help understand their microbial diversity and to identify beneficial bacterial species for improving quality.     

Age and growth of four Sardinella species from Sri Lanka

Age, growth and spawning season were studied in Sardinella sirm, S. albella, S. gibbosa and S. longiceps collected by gill nets in the coastal waters of Sri Lanka.Ageing was based on primary growth rings in the otoliths. The daily periodicity as verified for S. sirm by comparing mean increase in number of rings and the time elapsed between a series of sampling dates. For the other species, the same periodicity was inferred from the similarity in structure to that of S. sirm.The species reach the following lengths after 1 year: S. albella, 12 cm; S. gibbosa, 13 cm; S. longiceps, 14 cm; S. sirm, 20 cm. The total life-cycles are 1–1.5 years.Apparently, all species reach maturity after about 1 year. There seem to be two spawning seasons per year. Estimates based on counting growth rings indicate that these coincide with the south west and north east monsoonal changes. This also agrees with the changes in maturity stages observed.     

Comparative effects of diets consisting of one or two algal species upon assimilation efficiencies and growth of juvenile oysters, Crassostrea virginica (Gmelin)

Juvenile oysters, Crassostrea virginica, were fed one of six diets consisting of three species of algae fed singly or in two-species combinations. Dietary components were Isochrysis galbana (Diet I), Platymonas suecica (Diet P), and Thalassiosira pseudonana (Diet T). High (Diet TI), intermediate (Diets T, I, PT, PI), and low (Diet P) growth responses were observed. Intermediate and low growth responses were explained in terms of differing levels of ingestion and assimilation. The high growth response with Diet TI was not explained by measured values of ingestion and assimilation.     

Susceptibility by amberjack (Seriola dumerili), yellowtail (S. quinqueradiata) and Japanese flounder (Paralichthys olivaceus) to Neobenedenia girellae (Monogenea) infection and their acquired protection

This study investigated: 1) susceptibility differences to infection by Neobenedenia girellae (Capsalidae) between amberjack Seriola dumerili (Carangidae), yellowtail S. quinqueradiata and Japanese flounder Paralichthys olivaceus (Paralichthyidae); 2) growth and egg production of N. girellae on each fish species; 3) acquired protection of each fish species against this parasite. The number of N. girellae on S. dumerili was significantly higher than on S. quinqueradiata and P. olivaceus when these fishes were exposed to oncomiracidia in the same aquarium. Neobenedenia girellae growth on S. dumerili was fastest and, thus the number of eggs laid by parasites on S. dumerili was greater than on the other two species. Seriola dumerili and P. olivaceus, which were previously infected with N. girellae and treated by freshwater bath, acquired partial protection against re-infection by N. girellae. The relative re-infection of three S. dumerili individuals out of eleven individuals was markedly low compared with the initial infection, and the relative initial infection and re-infection on two P. olivaceus out of eleven individuals was markedly low. The results of this study could be useful to control N. girellae infections when cultivating S. dumerili, S. quinqueradiata and P. olivaceus.     

不同地理种群瓦氏马尾藻ITS序列特征及其系统进化分析

为研究不同地理种群瓦氏马尾藻ITS的序列变异,实验采用PCR扩增和序列测定方法,获得了3个不同地理种群15株瓦氏马尾藻的ITS全长序列,并进行序列分析。结果显示,15个个体共出现4种不同的ITS序列,它们相应的ITS1、5.8S、ITS2长度相同,均为762、158和507 bp,共有3个位点发生碱基变异。结合从GenBank中下载的马尾藻科3个属24种马尾藻的ITS序列,以羊栖菜属的羊栖菜,喇叭藻属的拟小叶喇叭藻和下延喇叭藻作为3个外群,采用邻位相连法构建分子系统进化树,结果显示,瓦氏马尾藻的ITS序列优先聚在一起然后以较高的置信度与南海马尾藻和球囊马尾藻聚为一支,在系统发生上显示出更近的亲缘关系。在选取的马尾藻中瓦氏马尾藻与南海马尾藻遗传距离最近为0.004,与拟小叶喇叭藻遗传距离最远为0.422。     

Effect of algal diet and temperature on the biochemical composition of the rotifer, Brachionus plicatilis

Rotifers were fed on four species of unicellular algae (Dunaliella tertiolecta, Pavlova lutheri, Phaeodactylum tricornutum and Isochrysis galbana) at a range of temperatures. Measurements were made of growth rate, weight and level of total protein, carbohydrate and lipid. Large differences were found in rotifer weight and biochemical composition as a result of feeding and starvation. There were only minor differences attributable to the type of alga used as a food.     

Predation by neogastropods on <Emphasis Type="Italic">Turbo cornutus</Emphasis> juveniles and other small gastropods inhabiting coralline algal turfs

To understand the effects of predation by gastropods on juveniles of the Japanese spiny turban snail Turbo cornutus, a field survey and laboratory experiments were conducted. The species compositions of the order Neogastropoda inside turfs of articulated coralline algae (ACA) on the east coast of Sagami Bay were surveyed monthly, and the dominant species inside the ACA turfs were identified. Laboratory experiments were conducted to examine the effects of predation by the dominant neogastropods on small gastropods in ACA turfs. Turban snails, which inhabit ACA turfs during their juvenile stages, were predated by two muricid gastropods, Ergalatax contractus and Thais bronni, with the predation rate of E. contractus being significantly higher than that of T. bronni. While E. contractus could also predate two other species of gastropods, Anachis misera and Cantharidus japonicus, the predation rates on these two snails were significantly lower than on T. cornutus. The observed defensive behaviors were different among the three prey species, and the defensive strategy related to the calcified operculum of T. cornutus was not efficient against predation by E. contractus. It is concluded that the predation by E. contractus represents an important factor in the mortality of juvenile turban snails inside ACA turfs.     

Ontogenetic habitat and dietary shifts in Japanese turban snail <Emphasis Type="Italic">Turbo cornutus</Emphasis> at Nagai,Sagami Bay,Japan

A field survey and a laboratory experiment were conducted to examine ontogenetic shifts in habitat and diet of the turban snail Turbo cornutus. The main habitat of turban snail juveniles smaller than 10 mm shell height (SH) was turfs of articulated coralline algae, and that of adults larger than 50 mm SH was kelp beds of Ecklonia bicyclis and Ecklonia cava. However, the ontogenetic habitat shift during the juvenile stage of 20–50 mm SH was unclear. From the results of a long-term feeding experiment and stable isotope analysis, the gelidiacean alga Gelidium elegans was assumed to be more important as a food source for juvenile turban snail than E. cava in the field. However, the frequency of individuals inhabiting gelidiacean algal turfs was low in both juvenile and adult stages. Thus, the ontogenetic shifts in habitat and diet did not coincide and other factors, i.e., daytime refuge, are important. As the growth rate was higher in the juveniles fed on the two algal species than those fed on single algal species, co-occurrence of various algal habitats in rocky shore ecosystems as a coastal ecosystem complex may enhance growth of turban snail juveniles feeding on a combination of different algal species.     

Genetic identification and morphology of naturally spawned eggs of the Japanese eel Anguilla japonica collected in the western North Pacific

Eggs of the Japanese eel Anguilla japonica collected in the western North Pacific were identified by onboard species-specific polymerase chain reaction (PCR) and DNA nucleotide sequencing after the cruise. Fish eggs of various species were collected by large plankton net tows at 12 stations along the southern part of the West Mariana Ridge on 19–25 May 2009. A total of 43 fish eggs were distinguished morphologically as possibly being of A. japonica. Thirty-one of those were analyzed by PCR, which included 15 eggs collected at 12°50–55′N, 141°15–20′E (in 5 tows) that showed positive results. The 16S ribosomal RNA (rRNA) gene sequences of eggs determined after the cruise indicated that 31 A. japonica eggs had been collected. The remaining eggs were of mesopelagic eel species (Serrivomeridae and Derichthyidae), or unidentified species. The morphological characteristics of the A. japonica eggs were consistent with those of artificially spawned eggs, except they had a slightly larger diameter. The egg diameter range did not overlap with those of mesopelagic eels of the Serrivomeridae, which often spawn in the same area as A. japonica. These results suggest that egg diameter and embryo shape can be used to morphologically identify naturally spawned A. japonica eggs.     

Influence of the dietary inclusion of <Emphasis Type="Italic">Gracilaria cornea</Emphasis> and <Emphasis Type="Italic">Ulva rigida</Emphasis> on the biodiversity of the intestinal microbiota of <Emphasis Type="Italic">Sparus aurata</Emphasis> juveniles

Partial substitutions of fish meal by 5, 15, or 25 % of Gracilaria cornea or Ulva rigida in experimental diets were evaluated to study their effects on biodiversity of intestinal microbiota composition in gilthead seabream (Sparus aurata) juveniles. The diets were offered to duplicate groups of 15 juvenile fish (14.0 ± 0.5 g) for 70 days, and at the end of the experiment the intestinal microbiota from four specimens of each treatment was analysed by denaturing gel gradient electrophoresis. Results showed that the substitution of fish meal by algae meal induced important modifications in the intestinal microbiota community, as a big reduction of the biodiversity when the highest percentage (25 %) of U. rigida was included. On the contrary, an increase on the number of species was detected when a 15 % of algae was included. Various Lactobacillus delbrueckii subspecies were selectively stimulated when G. cornea was included in the feed, and other bacterial species, such as those included in the Vibrio genus, were reduced.