Ingestion,fecundity and population growth of Harpacticus sp. (Harpacticoida,copepod) fed on five species of algae

We investigated the ingestion, fecundity and population growth of Harpacticus sp. fed on monodiet or mixed diets to evaluate the effects of different algae on Harpacticus sp. Harpacticus sp. fed on diatoms (Skeletonema costatum and Chaetoceros curvisetus) had higher ingestion and pellet production. Time to attain 100% in proportions of gravid females differed, with quickest to slowest: S. costatum, C. curvisetus, Gymnodinium sp. and Heterosigma akashiwo, with the exception of Prymnesium parvum (≤16.67%). S. costatum or C. curvisetus produced higher populations than the other three diets, supported complete development to adulthood, and resulted in doubling copepod population within four days, while no population growth occurred for the other three diets. Mixed‐diet experiments showed that egg production and gross growth efficiencies reduced significantly when fed on H. akashiwo, Gymnodinium sp. or P. parvum mixed with Isochrysis galbana, in comparison with 100% I. galbana. Thus, S. costatum and C. curvisetus were beneficial foods while the other three diets were potentially toxic for Harpacticus sp. The data in this article provide further recognition of nutrient deficiency or toxicity of different algae on copepods.     

龙须菜苯丙烷类抑藻活性物质的分离纯化及其对6种赤潮微藻的抑藻活性

采用溶剂浸提、液液萃取、硅胶柱层析和硅胶薄层层析等分离方法以及红外光谱、质谱和核磁共振谱等光谱技术,初次从龙须菜中纯化得到4种苯丙烷类化合物:邻苯二丙酸、gossonorol、7,10-epoxy-ar-bisabol-11-ol和对羟基苯乙醇,并进一步分析了此4种苯丙烷类化合物对强壮前沟藻、赤潮异弯藻、米氏凯伦藻、球形棕囊藻、东海原甲藻和中肋骨条藻生长的影响。结果表明,它们对以上6种赤潮微藻的生长表现出明显的选择性抑制作用。其中,邻苯二丙酸、gossonorol和对羟基苯乙醇表现出更为广泛的抑藻活性。比较此4种苯丙烷类化合物和重铬酸钾对赤潮微藻生长的半抑制效应浓度EC50-96 h,发现邻苯二丙酸对强壮前沟藻、赤潮异弯藻和球形棕囊藻,gossonorol和羟基苯乙醇对赤潮异弯藻和球形棕囊藻,7,10-epoxy-ar-bisabol-11-ol对赤潮异弯藻和米氏凯伦藻在生长抑制方面比重铬酸钾更具有优势。     

浒苔对赤潮异湾藻的克生作用

研究了浒苔对赤潮微藻—— 赤潮异湾藻生长的克生效应。结果表明, 浒苔新鲜组织对赤潮异湾藻的生长具有强烈的克生效应; 一次性添加浒苔培养过滤液对赤潮异湾藻生长没有显著的抑制作用, 而半连续添加浒苔培养过滤液对赤潮异湾藻生长具有显著的克生作用; 浒苔干粉末和甲醇抽提液均对赤潮异湾藻的生长产生显著的抑制作用。试验结果表明, 抑制赤潮异湾藻生长的物质很可能更多地存在于浒苔组织内, 向环境中分泌的克生物质较少。因而, 克生物质的连续分泌是有效克制赤潮异湾藻生长的关键。同时, 克生作用存在着明显的浓度效应, 当抑制物浓度达到一定阈值才表现出明显的克生作用, 在阈值之上浓度越高克生作用越强。     

Species diversity of the marine diatom genus Skeletonema in Japanese brackish water areas

The genus Skeletonema includes phytoplankton species that are important primary producers in marine food chains. Brackish waters have been reported to be one of the important habitats of some species of Skeletonema. To elucidate the species diversity of Skeletonema in brackish waters, we investigated three Japanese brackish bodies of water: the coastal waters of Toyama Bay, a tidal area of the Chikugo River, and a constructed reservoir in Isahaya Bay. We used molecular analysis based on large subunit rDNA and fine morphological structure to identify species. Skeletonema costatum s.s. (sensu stricto) was isolated at salinities as low as 0.6, but Skeletonema dohrnii, Skeletonema subsalsum, and Skeletonema tropicum were not found at salinities below 11.0. S. costatum s.s. could survive transfer from a medium with a salinity of 15 to a salinity of 2, but S. dohrnii did not survive in the same experiment. Only S. costatum s.s. germinated from the sediment of a reservoir in which the salinity was 0.1–1.4; incubation conditions included temperatures of 10, 15, 20, 25, and 30 °C and salinities of 5 and 30. Skeletonema costatum s.s. was identified as the species most adaptable to low-level salinity variations throughout its lifecycle.     

Isolation and characterization of Noctiluca-killing bacteria from a shrimp aquaculture pond in Thailand

To control harmful algal blooms (HABs), in particular dinoflagellate Noctiluca scintillans, which causes damage to shrimp production, algicidal bacteria were isolated from shrimp pond water and screened for their killing effect against N. scintillans under laboratory condition. Among 260 bacterial isolates, 10 showed killing effects on N. scintillans. Out of 5 strains showing relatively stronger killing activity, 4 strains belonged to the Gammaproteobacteria group. Among these, strain BS2 (identified as Marinobacteria salsuginis) showed the strongest Noctiluca-killing activity. In the present study, we also investigated bacterial effects against growth of other phytoplankton species. Strain BS2 killed only N. scintillans but not any other phytoplankton (Heterosigma akashiwo NIES-6, Chatonella antique NIES-1, Chaetoceros ceratosporum, Prorocentrum lima, and Dunaliella sp.). Results of the present study lead us to expect prevention of damage by HABs, in particular dinoflagellate N. scintillans, in shrimp farming and mitigation for shrimp production in ponds using Noctiluca-killing bacteria.     

Evaluation of microalgae diets for Litopenaeus vannamei larvae using a simple protocol

We evaluated native marine microalgae isolated from NortheasternVenezuela, as food for Litopenaeus vannamei larvae,compared to six microalgae commonly used in aquaculture. We evaluated themicroalgal ingestability calculating the biomass consumed by larvae, later weevaluated three monoalgal diets on protozoea stages of L.vannamei over 72 hr based on the larval biomass,RNA/DNAindex and the proportion of stage retarded larvae. The presence of everymicroalgae species in the protozoea's digestive tract indicated theirconsumption. Chaetoceros spp. showed high ingestabilityandTetraselmis spp. and Skeletonema sp.Ch1 showed low ingestability. The native microalgaeChaetoceros sp. A1 and Chaetocerossp.G1 were evaluated as monoalgal diets compared to Chaetocerosgracilis. Ch. gracilis demonstrated the bestresults when the evaluation was performed with the RNA/DNA index. However, thesurvival rate, larval biomass and percentage of stage retarded larvae fed withChaetoceros sp. G1 were not statistically different fromthose fed with Ch. gracilis. Taking into account thatnative microalgae produced higher biomass in shorter times when they werecultured in field conditions, the results suggest that native microalgae areadequate for large-scale culture conditions, thus reducing costs. Thedetermination of microalgal size and the larval consumption, dry and organiclarvae biomass and specially the RNA/DNA index, were valuable features of thisevaluation. The protocol developed in this study could be used to evaluatemicroalgae quality under different environmental conditions and productionregimes.     

Salinity Preference of Two Diatoms and Their Growth Performance in Three Prepared and Two Alternative On-Farm Media Sources

ABSTRACT

Salinity and preferred nutrient composition of Chaetoceros affinis and Skeletonema costatum were tested. Salinity was tested at 12, 16, 23, 28, and 32 ppt. Three compositions of nitrogen (N), phosphorus (P), and potassium (K) and two on-farm low-cost alternative media sources, cow urine (CU) and hatchery waste water (HWW), were tested to find the suitable NPK ratio and to determine the efficacy of the on-farm media respectively. Chaetoceros affinis preferred 23 ppt salinity, while Skeletonema costatum preferred both 28 and 32 ppt. Among the NPK treatments, higher cell density was observed in 4:1:1 followed by 4:2:1 ratios. Significant differences in maximum cell density of both species were observed in the NPK treatments while only S. costatum showed significant difference in CU and HWW.     

珠江口海域双胞旋沟藻赤潮对卤虫幼体、鱼苗和虾苗的急性毒性

为研究2009年10月下旬在广东珠海海域爆发的双胞旋沟藻赤潮对养殖鱼类及水体中其它生物的影响,实验以卤虫幼体、金鼓鱼苗和凡纳滨对虾苗作为受试生物,在赤潮现场测试双胞旋沟藻对卤虫幼体、鱼苗和虾苗的急性毒性效应。结果显示,24 h双胞旋沟藻对卤虫幼体的LC₅₀(半致死浓度)为9.55×10⁴/mL,藻密度为2.5×10³/mL的双胞旋沟藻对卤虫幼体的LT₅₀(半致死时间)为48.5 h。60 h内该赤潮水体对鱼苗和虾苗的存活无不利影响,卤虫幼体和金鼓鱼苗均可摄食双胞旋沟藻,卤虫幼体对双胞旋沟藻的摄食率低。研究表明,双胞旋沟藻赤潮水体对卤虫幼体有一定的毒性作用,但在低藻密度条件下,卤虫幼体能以该藻为食并维持其生命,双胞旋沟藻对金鼓鱼苗和凡纳滨对虾苗无急性毒性作用。     

Selectivity of the rotifer Brachionus plicatilis fed mixtures of algal species with various cell volumes and cell densities

The ability for food selectivity of rotifer Brachionus plicatiliswas studied in the laboratory by feeding mixtures of microalgae with various cell volumes. Chlorella sp. (≈22 µm³) was the reference algal species, and Asteromonas gracilis, (Chlorophyta) (≈2150 µm³), Tetraselmis suesica (≈268 µm³), Dunaliella salina (≈52 µm³) and Chaetoceros sp. (≈150 µm³) the experimental species. Each was mixed with Chlorella and fed in three experiments. In the first experiment, filtration and ingestion rates of rotifers each fed with algae revealed that the highest values were measured with the mixture of Chlorella + Asteromonas, and the lowest for Chlorella + Chaetoceros. In the second and the third experiments, by using several combinations of algal densities with the mixture of Chlorella + Asteromonas, a selectivity ability of the rotifers for Asteromonas was found. A hypothesis is presented that accounts for the preference of rotifers for Asteromonas, which is suggested as a new candidate species for use in live food production of fish hatcheries.     

Drinking behaviour in sea water and fresh water teleosts,the role of the renin-angiotensin system

Basal drinking rate and responses to administered angiotensin were examined in 12 species of fish. The responses of representative euryhaline, stenohaline marine and fresh water species to pharmacological manipulation of endogenous renin-angiotensin system (RAS) activity were also investigated.Basal drinking rates were consistently low in stenohaline and euryhaline fresh water fish, and all species examined showed an increased imbibition in response to administered angiotensin. Marine fish drank large volumes of water, rates varying considerably between species, with euryhaline species exhibiting lower rates than stenohaline groups. The extremely high drinking rates observed in the sea scorpion were associated with a high plasma osmolality. With the exception of the sea scorpion, all other species examined in sea water showed a further rise in drinking in response to exogenous angiotensin.Although the freshwater stenohaline carp showed a dipsogenic response to angiotensin, it was apparently unable to evoke this response when fish were acclimated to brackish water. The high drinking rates of both euryhaline and stenohaline fish held in sea water appeared dependent upon an activated endogenous RAS, and were lowered following inhibition of Al to All conversion by Captopril. Drinking was further stimulated in these marine species following stimulation of endogenous RAS activity by the administration of the hypotensive agent Papaverine. The study endorses a role for the RAS in the control of adaptive drinking in euryhaline and stenohaline marine teleosts.     

Factors affecting the large-scale production of four species of commercially important marine algae

Semi-continuous cultures of four algal food species were maintained in 80 l and 200 l vessels internally illuminated by six 80 W fluorescent lamps. At standard densities Skeletonema costatum produced the greatest average yield, equivalent to 120 l d^?1 for 7–8 weeks. Chaetoceros calcitrans, Isochrysis aff. galbana (clone T. Iso.) and Chroomonas salina produced 50–60 l d^?1 for 2–4 weeks. Yields for each algal species were similar from 80 l and 200 l vessels and lower when four 80 W lamps were used. The harvesting regime to achieve the above yields is evaluated.     

Influence of different diets on the growth,survival, fecundity and proximate composition of brine shrimp Artemia franciscana (Kellog, 1906)

Artemia franciscana was studied under in vitro conditions, and fed with five different microalgae species, two organic diets and one mixed diet to evaluate the nutritional profile and growth performance. The A. franciscana instar‐I nauplii were stocked at a density of 100 per litre of ground borewell water with a salinity of 70 ppt in a plastic container and the culture continued for 12 days. The A. franciscana was fed with five microalgal species, such as Tetraselmis sp., Chaetoceros sp., Isochrysis sp., Thalassiosira sp. and Nannochloropsis sp., and organic diets of rice bran, soybean meal, and a mixture of Tetraselmis sp. and Chaetoceros sp. with rice bran, giving a total of eight experiments. The growth of A. franciscana was measured by length, and was observed to be the greatest (10,850 μm) in the group that was fed the mixed algae combination, and the lowest (7,290 μm) in the group that was fed Nannochloropsis sp. Among the different treatments, the highest survival value of 76% was seen in the group that was fed Chaetoceros sp. The fecundity was higher in A. franciscana that were fed with algae Chaetoceros sp., while the lower rate was observed with the group that was fed Nannochloropsis sp. Analysis of proximate composition showed increased levels in the animals fed with Chaetoceros sp. (lipid, 19.40%; eicosapentaenoic acid, 6.70 mg; docosahexaenoic acid, 10.20 mg; amino acids: proline, 2.06 μg; histidine, 0.77 μg, arginine, 0.65 μg, glycine, 0.80 μg, glutamic acid, 1.75 μg; threonine, 1.42 μg) and the mixed diet (protein, 58.59%; carbohydrate, 21.30%; amino acids: asparagine, 0.90 μg; serine, 1.65 μg; tryptophan, 1.66 μg; leucine, 1.50 μg; phenylalanine, 1.14 μg; valine, 0.66 μg). All components showed the lowest level in the diet fed with Tetraselmis species. These A. franciscana populations showed better survival, higher fecundity and biochemical profiles when cultured with microalgae Chaetoceros sp. These data are useful to improve A. franciscana culture in aquaculture to produce quality cysts and biomass, especially in feeding larvae of marine species.     

鲫肠道乳酸菌的分离及益生特性

为获得鱼源益生乳酸菌,本研究从健康鲫肠道内分离鉴定得到38株乳酸菌,并选择其中8株乳酸菌进行体外益生特性分析。结果显示,8株乳酸菌均能在p H=4.5和10%鲫胆汁环境中存活,对嗜水气单胞菌、豚鼠气单胞菌、无乳链球菌和副溶血弧菌均有较强的抑菌能力,但菌体表面疏水性和自凝集能力具有菌株特异性。选择5株疏水性较高(63%~89%)、自凝集能力强(37%~45%)的乳酸乳球菌进行体外黏附能力测定,结果显示,黏附能力在菌株间具有差异性(4.5%~7.9%),但均能显著降低嗜水气单胞菌NJ-35的体外黏附率,黏附抑制率达30%~35%;最后对筛选出的3株高黏附力的乳酸乳球菌进行安全性评价,发现3株菌对鲫均无致病力。鱼源乳酸菌的筛选,为鲫养殖生产中益生菌的实际应用提供了理论依据。     

Digestibility of the defatted microalgae Nannochloropsis sp. and Desmodesmus sp. when fed to Atlantic salmon,Salmo salar

The aim of the study was to investigate the apparent digestibility coefficient (ADC) of defatted biomass derived from microalgae Nannochloropsis sp. and Desmodesmus sp. when fed to Atlantic salmon postsmolts in seawater. Two experiments, one employing cold‐pelleted and the other employing extruded pellets, were carried out to determine the ADC of dry matter (DM), protein, ash and energy. The test feeds consisted of a fishmeal‐based reference feed and the algal biomass, at a ratio of 70:30, and yttrium oxide as the inert marker. The ADC of DM and protein in fish fed microalgae‐incorporated cold‐pelleted feeds were significantly higher for Nannochloropsis sp. compared with Desmodesmus sp. The Nannochloropsis sp. inclusion in extruded feeds also led to higher ADCs of DM and energy compared with Desmodesmus sp. The extrusion processing significantly increased the ADC of ash in both defatted microalgae biomass, as well as ADCs of DM in Nannochloropsis and protein in Desmodesmus sp. In conclusion, the microalga Nannochloropsis sp. was more digestible than Desmodesmus sp., and extrusion processing can be used as a means to improve digestibility of certain nutrients.     

The use of Artemia sp. or mysids as food source for hatchlings of the cuttlefish (Sepia officinalis L.); effects on growth and survival throughout the life cycle

Twoexperiments were conducted to determine the effects ofArtemia sp. or mysids on growth and survival ofS. officinalis hatchlings, and their effect throughout thelife cycle. For experiment I, for the first 20 days, one group was fed adultArtemia sp. and the other was fed mysid shrimp(Paramysis nouvelli). Eggs laid by females in both groupswere counted and weighed, and hatchlings were weighed, to determine differencesin both groups. For experiment II, during the first 10 days, one group was fedArtemia sp. and the other was fed mysids (P.nouveli). After the period of differentiated feeding, the 2 groupsinexperiment I were fed grass shrimp (Paleomonetes varians)to 70 days old, and dead crabs (Carcinus maenas)afterwards. Cuttlefish in experiment II were fed grass shrimp from day 10 untilthe end of the experiment. For both experiments, hatchlings fed mysids grewsignificantly bigger (p < 0.01) and survival was higher. For experiment I,eggs laid by females fed mysids and the hatchlings born from these eggs werebigger (p < 0.001) compared to the group fed Artemiasp.initially. Individual fecundity was slightly higher for females in the groupfedArtemia sp. (163 eggs female^–1) than forthe group fed mysids (144 eggs female^–1). Egg laying startedatthe age of 125 days and lasted 45 days in both groups. Time between first egglaying day and first hatchlings to be born was 21 days. The last female to die(after spawning) in both groups was 167 days (less than 6 months old).     

Mutualism between euryhaline tilapia Sarotherodon melanotheron heudelotii and Chlorella sp.—Implications for nano-algal production in warmwater phytoplankton-based recirculating systems

The West-African euryhaline tilapia, Sarotherodon melanotheron heudelotii shift from visually feeding on zooplankton when juveniles to mostly filter feeding on phytoplankton when adults. When reared using an appropriate ration in intensive aquaculture systems, S. m. heudelotii also consume algal-based detritus, and contribute to sediment mineralization, clean up their environment, and ultimately stimulate and sustain algal growth. We analysed such practical advantages for phytoplankton-based recirculating systems, using S. m. heudelotii and Chlorella sp. as biological material originating from the prototype of such a system operated in Senegal. We performed a 24-h factorial design experiment in 36 tubs, cross-classifying three levels of S. m. heudelotii (fishless control, unfed fish, and fed fish) with four levels of Chlorella initial density.Chlorella overall mean density increased significantly from fishless, to unfed fish, and fed fish treatments, and with Chlorella initial density. S. m. heudelotii did not alter nitrogen nor phosphorus concentrations, only affected by algal initial densities. Most ammonia excreted by fish was probably uptaken by Chlorella. Bacteria-mediated diurnal nitrification was possibly an alternative ammonium loss mechanism at highest oxygen concentrations. Algae were not limited by nitrogen or phosphorus but most likely by low dissolved organic carbon availability. Chlorella differential responses with fed vs. unfed Sarotherodon suggest that CO₂ supplied by heterotrophic S. m. heudelotii respiration played a key role. Observed Chlorella growth rates were similar to the highest rates obtained in algal mass cultures, enriched with CO₂, nitrate and phosphate, under artificial lighting.Our results suggest the existence of a Sarotherodon-Chlorella mutualism in our systems, where S. m. heudelotii provide CO₂, the major limiting factor of Chlorella growth, whereas Chlorella oxygenate and detoxify the water media from ammonia, promoting S. m. heudelotii production. This mutualism could be used to optimize photosynthetic suspended-growth aquaculture systems, particularly in the Tropics where light is abundant and temperature is continuously high.     

Evaluation of three phytoplankton species as food for the pearl oyster <Emphasis Type="Italic">Pinctada fucata</Emphasis>

In the pearl cultivation farms of the Ehime Prefecture, Japan, mass mortalities of the pearl oyster Pinctada fucata have occurred since 1994. The occurrences of mass mortality roughly coincided with a shift of the dominant phytoplankton from Skeletonema and Chaetoceros to Chaetoceros and Nitzschia all of which belong to Bacillariophyceae. Hence, we evaluated Nitzschia, together with Chaetoceros and Isocrysis, as food for the oyster. Wet weights, lengths, widths, glycogen contents, and growth rates in terms of wet weight of the oysters in all the feeding treatments were significantly higher than those in the non-feeding treatment. The highest glycogen content (2.34%) and growth rate (2.21 g month⁻¹) were found in the Chaetoceros treatment. Growth rate in the Isocrysis treatment (1.63 g month⁻¹) was also high, although glycogen content in this treatment (0.41%) was low. In the Nitzschia treatment, growth rate of the oyster (0.94 g month⁻¹) was the lowest and glycogen content (0.83%) was also low relative to that in the Chaetoceros treatment. Chlorophyll a concentration in fecal pellets was lowest in the Nitzschia treatment (<2.7 μg mg⁻¹), suggesting more complete digestion of Nitzschia by the oyster. Thus, Nitzschia was edible and digestible but not assimilated by P. fucata. We propose the following scenario for the relationship between Nitzschia dominance and mass mortality. When Nitzschia dominates in a culture area, the physiological condition of P. fucata deteriorates due to low assimilation of Nitzschia by the oyster, followed by susceptibility of the oyster to infection by agents lethal to the oyster.     

Antifreeze proteins in the urine of marine fish

Several species of marine teleosts have evolved blood plasma antifreeze polypeptides which enable them to survive in ice-laden seawater. Four distinct antifreeze protein classes differing in carbohydrate content, amino acid composition, protein sequence and secondary structure are currently known. Although all of these antifreezes are relatively small (2.6–33 kd) it was generally thought that they were excluded from the urine by a variety of glomerular mechanisms. In the present study antifreeze polypeptides were found in the bladder urine of winter flounder (Pseudopleuronectes americanus), sea raven (Hemitripterus americanus), ocean pout (Macrozoarces americanus) and Atlantic cod (Gadus morhua). Since the plasma of each of these fish contains a different antifreeze class it would appear that all four classes of antifreeze can enter the urine. The major antifreeze components in the urine of winter flounder were found to be identical to the major plasma components in terms of high performance liquid chromatography retention times and amino acid composition. It is concluded that plasma antifreeze peptides need not be chemically modified before they can enter the urine.     

Sprinkling: a new method of distributing live algae food in marine coastal ponds used for Manila clam Tapes philippinarum (Adams & Reeve) intensive culture

Sprinkling, an agricultural irrigation technique, has been tested for the intensive culture of Manila clams up to marketable size (35-50 mm) in coastal ponds 400 m² in area. The sprinkling system is used for the distribution of seawater and live algal food Skeletonema costatum (Grev.) Clev. Sprinkling is a simple, low cost and efficient method of food distribution which resolves the major problem of decreasing bivalve growth associated with single point water entry in small-scale raceways. Uniform growth obtained with sprinkling is principally due to an even scattering of food over the whole surface area of large-scale ponds.     

血卵涡鞭虫单克隆抗体的制备与初步应用

用血卵涡鞭虫可溶性抗原免疫BALB/c小鼠,经常规融合、间接ELISA方法筛选,将所得阳性克隆再经3次亚克隆后,共获得3株针对血卵涡鞭虫的单克隆抗体(2B₂、3G₄、4G₇),单克隆抗体亚类鉴定表明,三者为IgG类抗体。用筛选的杂交瘤细胞株制备小鼠腹水抗体,其细胞上清及腹水效价分别为5.12×10^-4和8.00×10^-4。进一步利用单克隆抗体建立间接荧光抗体方法对单抗特异性进行鉴定,阳性虫体被染上黄绿色荧光,而正常梭子蟹血淋巴则未被染色。用单克隆抗体和多克隆兔抗血清以羊抗鼠HRP-IgG为酶标抗体,建立了检测血卵涡鞭虫的双抗体夹心ELISA方法,该方法对血卵涡鞭虫阳性标本检测符合率为100%。结果表明,制备的单克隆抗体效价高、特异性好,可用于血卵涡鞭虫的早期临床诊断。