Effects of chloramphenicol, erythromycin, and furazolidone on growth of Isochrysis galbana and Chaetoceros gracilis

This study focused on determining the effects of antibiotics on microalgae used as food for scallop larvae. Six different dose levels of chloramphenicol, erythromycin, and furazolidone were added to cultures of Isochrysis galbana and Chaetoceros gracilis. An in vivo experiment was subsequently conducted to determine the effect of chloramphenicol and erythromycin on larval survival of the Pacific calico scallop Argopecten ventricosus in tanks and on the population of its associated bacteria. Results showed that growth of I. galbana was not significantly affected by chloramphenicol or erythromycin at the test doses of 0.5, 1.0, 3.0, 6.0, 9.0, and 12.0 mg/l. C. gracilis was significantly sensitive to erythromycin and chloramphenicol at doses higher than 0.5 and 3.0 mg/l, respectively. Furazolidone inhibited the growth of both I. galbana and C. gracilis at all test doses. Results showed that exposure of scallop larvae to a dose of 6 mg/l chloramphenicol or erythromycin did not significantly affect growth of I. galbana, significantly enhanced survival of the scallop larvae, and inhibited the growth of Vibrio spp. in tanks. This study demonstrated the adverse effect of chloramphenicol, erythromycin and furazolidone on I. galbana and C. gracilis microalgae but the positive effect on survival of the scallop larvae, decreasing associated bacterial population.     

The effect of diet type and quantity on the development of common sea urchin larvae Echinus esculentus

Several species of sea urchins are now being cultivated for commercial purposes and with the continued increased demand for sea urchin gonads as a food product, new species are being assessed for their aquaculture and market potential. This study focussed on establishing protocols for the production of common sea urchin Echinus esculentus larvae and juveniles to assess its potential as an echinoculture species. Two trials were carried out, the first trial evaluated the influence of three microalgal diets (D=Dunaliella tertiolecta only, mixed D/P=D. tertiolecta plus Phaeodactylum tricornutum and P=P. tricornutum only) on larval morphology. Larval length, width, post-oral arm length and rudiment length were significantly effected by diet. Diets D and D/P prompted more rapid metamorphosis. In the second trial, the effects of different rations of D. tertiolecta were tested. The food ration, standard ration (SR; 1000, 3000, and 5000 cells ml⁻¹) and high ration (HR; 3000, 9000, and 15,000 cells ml⁻¹) were increased as the larvae acquired the 3rd and 4th pair of larval arms. Larvae fed the SR were significantly larger (longer and wider) and had significantly longer rudiments than those in the HR treatment. The number of larvae metamorphosing and settling onto substrates was significantly higher in treatment SR compared to HR. Optimising the larval diet shortened the larval stage from 21–23 days in the first trial to 16 days in the second trial. The maximum percentage of metamorphosing individuals which survived to post-larvae or juveniles (10 days after they were first judged competent to settle) was 46.6%, suggesting E. esculentus is a viable aquaculture candidate.     

Effect of different permeable and non-permeable cryoprotectants on the hatching rate of rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>) embryos

The purpose of the present study was to investigate the toxicity of cryoprotectants on the hatching rate of rainbow trout (Oncorhynchus mykiss) embryos. Epiboly and first eye pigmentation stage embryos were immersed in six permeable cryoprotectants, dimethyl sulfoxide (DMSO), glycerol (Gly), methanol (MeOH), propylene glycol (PG), ethylene glycol (EG), and acetamide (Ac), in concentrations of 1–5 M for 5 or 10 min and two non-permeable cryoprotectants, sucrose (Suc) (10%, 15%, 20%) and polyvinyl pyrrolidone (PVP) (5%, 10%, 15%) for 5 min. The embryos were then washed and incubated until hatching occurred. The toxicity of the cryoprotectant was assessed by the hatching rate. The results illustrated that permeable cryoprotectant toxicity for rainbow trout embryos increased in the order of PG < DMSO < MeOH < Gly < EG < Ac. The hatching rate of the embryos treated with permeable cryoprotectants decreased (P < 0.05) with increased concentration and duration of exposure. There were no significant decreases in hatching rate of embryos treated with sucrose and PVP with the increase of concentration; sucrose had higher hatching rates than PVP. Rainbow trout embryos at first eye pigmentation stage exhibited greater tolerance to cryoprotectants than embryos at epiboly stage.     

Production of microalgal concentrates by flocculation and their assessment as aquaculture feeds

A novel technique was developed for the flocculation of marine microalgae commonly used in aquaculture. The process entailed an adjustment of pH of culture to between 10 and 10.6 using NaOH, followed by addition of a non-ionic polymer Magnafloc LT-25 to a final concentration of 0.5 mg L⁻¹. The ensuing flocculate was harvested, and neutralised giving a final concentration factor of between 200- and 800-fold. This process was successfully applied to harvest cells of Chaetoceros calcitrans, C. muelleri, Thalassiosira pseudonana, Attheya septentrionalis, Nitzschia closterium, Skeletonema sp., Tetraselmis suecica and Rhodomonas salina, with efficiencies ≥80%. The process was rapid, simple and inexpensive, and relatively cost neutral with increasing volume (cf. concentration by centrifugation). Harvested material was readily disaggregated to single cell suspensions by dilution in seawater and mild agitation. Microscopic examination of the cells showed them to be indistinguishable from corresponding non-flocculated cells. Chlorophyll analysis of concentrates prepared from cultures of ≤130 L showed minimal degradation after 2 weeks storage.

Concentrates of T. pseudonana prepared using pH-induced flocculation gave better growth of juvenile Pacific oysters (Crassostrea gigas) than concentrates prepared by ferric flocculation, or centrifuged concentrates using a cream separator or laboratory centrifuge. In follow up experiments, concentrates prepared from 1000 L Chaetoceros muelleri cultures were effective as supplementary diets to improve the growth of juvenile C. gigas and the scallop Pecten fumatus reared under commercial conditions, though not as effective as the corresponding live algae. The experiments demonstrated a proof-of-concept for a commercial application of concentrates prepared by flocculation, especially for use at a remote nursery without on-site mass-algal culture facilities.     

Energetic physiology of the Caribbean scallops Argopecten nucleus and Nodipecten nodosus fed with different microalgal diets

The Caribbean scallops Argopecten nucleus and Nodipecten nodosus are currently being placed into mass culture in Colombia. The limited availability of wild seed upon which to base these cultures has promoted research into the development of artificial production of this seed in hatcheries. In support of this effort, we studied the effects of different diets on the physiology of the two scallop species in order to determine the optimal feeding regimes for maintenance of adult specimens in the laboratory. Seven monoalgal diets were tested, including Chaetoceros calcitrans, Chaetoceros muelleri, Isochrysis galbana (Ig), Nannochloris oculata (No), Phaeodactylum tricornutum (Pt), Tetraselmis chui (Tc) and Tetraselmis tetrahele (Tt). Four mixed diets were also tested, including I. galbana + C. calcitrans (Ig + Cc), I. galbana + N. oculata (Ig + No), I. galbana + T. tetrahele (Ig + Tt), and I. galbana + lipid emulsion of docosahexanoic acid DHA (Ig + lip). All the dietary trials were carried out under uniform conditions of temperature (25 °C), salinity (36‰) and algal concentration (0.45 mg L^− 1). Physiological variables measured in association with each diet included feeding rates (clearance, ingestion and absorption), oxygen consumption and ammonium excretion rates as well as their scope for growth. The results showed that the best scope for growth for both scallops was obtained with diet Ig since this diet induced the highest feeding rates, accompanied by the lowest oxygen consumption and ammonium excretion. The feeding rates and scope for growth of A. nucleus were greater than those of N. nodosus for the majority of the diets, which was attributed to a higher rate of water pumping by the former species. Greater capacity for branchial food retention by A. nucleus was discarded as a possibility since N. nodosus had a greater branchial surface area per unit dry weight than A. nucleus. In spite of these differences, the oxygen consumption and the excretion rates were similar between the two scallops which suggested that A. nucleus was more efficient in its use of oxygen and retention of body proteins for physiological functioning. Mixed diets or addition of DHA did not permit increases in scope for growth in either of the scallops over that observed using monoalgal diet Ig, which suggest that biologically and economically this diet is optimal for the feeding of adult scallops in the laboratory.     

Viability of mass algal cultures preserved by freezing and freeze-drying

The microalgae Chaetoceros sp. and Phaeodactylum tricornutum were grown in semicontinuous 15 1 cultures, a volume commonly used for mid-scale inocula in aquacultural practices. The daily yields were concentrated and stored by freezing and freeze-drying for up to 1 month. After slow or fast freezing without the protective agents glycerol or Me₂SO, neither of the microalgae was capable of active growth, while with the cryoprotectants slow freezing gave good results after 7 days of storage at −20°C. After 15 and 30 days, however, microalgae growth was poor, probably due to bacterial contamination. Freeze-drying gave even worse results, which we attribute to severe losses of storage products after this treatment.     

溶藻细菌控藻作用及其在对虾养殖池塘中的应用前景

浮游微藻与养殖池塘的水质及对虾的健康状况密切相关,控制有害微藻的过度繁殖,维持优良的藻相对于养殖水环境的稳定、减少对虾疾病的发生具有重要意义。文章对浮游微藻与对虾养殖的关系及溶藻细菌的研究进展进行了概述,分析了溶藻细菌在对虾养殖中的应用前景,提出了利用溶藻细菌控制对虾养殖池塘有害微藻的研发思路。     

Preliminary studies on the cryopreservation of gilthead seabream (Sparus aurata) embryos

Vitrification could provide a promising tool for the cryopreservation of fish embryos. In order to achieve successful cryopreservation, several parameters should be taken into account in the design of a vitrification protocol. In the present study some relevant factors were investigated (permeable and non-permeable cryoprotectant toxicity, toxicity of vitrificant solutions, adequate container for embryo loading and temperature of thawing) using two gilthead seabream embryonic development stages (tail bud and tail-bud-free). Permeabilized embryos were incubated in dimethyl sulfoxide (DMSO), methanol (MeOH), ethylene glycol (EG) and 1,2-propanediol (PROH) in concentrations ranging from 0.5 to 6 M for 10 and 30 min and in 5%, 10% and 15% polyvinyl pyrrolidone (PVP), 10%, 15% and 20% sucrose or 0.1%, 1% and 2% X1000® for 2 min. After treatment, embryos were washed and incubated in seawater until hatched. The toxicity of permeable cryoprotectants increased with concentration and exposure time. EG was best tolerated by the embryos. Exposure to non-permeable cryoprotectants did not affect the hatching rate except at F stage. Six vitrificant solutions (DMSO—V1, V2 and V3 and EG—V1, V2 and V3) were tested using a stepwise incorporation protocol. The DMSO-based solutions contained 5 M DMSO + 2 M MeOH + 1 M EG plus 5% PVP, 10% sucrose or 2% X1000® and the EG-based solutions contained 5 M EG + 2 M MeOH + 1 M DMSO plus 5% PVP or 10% sucrose. Before loading the embryos into 0.5 ml straws or 1 ml macrotubes, toxicity tests were effected with these solutions. Our results demonstrated that DMSO-based solutions were better tolerated by seabream embryos than EG-based solutions. After thawing (water bath, 0 or 25 °C), embryos were evaluated by stereoscopic microscopy and the percentage of embryos with intact morphology was registered. The highest percentage of embryos with intact morphology (28%) was observed in samples frozen in macrotubes and thawed at 25 °C. Several malformations associated with ice crystal formation inside the embryos were detected. None of these embryos achieved hatching. Our results suggest that the absence of a proper incorporation of cryoprotectants prior to vitrification is the main problem that must be overcome. This procedure should be optimized in order to avoid ice crystal formation inside embryo compartments.     

PVP-碘对南美白对虾无节幼体的急性毒性试验

采用静水试验法，用PVP-碘对南美白对虾无节幼体进行了急性毒性试验。试验结果表明：PVP-碘对南美白对虾无节幼体的24h及40h半致死浓度分别为8.737mg/L及6.512mg/L，其安全浓度为0.65mg/L。     

Replacement of living microalgae with spray-dried diets for the marine mussel Mytilus galloprovincialis

Growth experiments were carried out in which juvenile mussels Mytilus galloprovincialis were fed diets composed of spray-dried algal products (Sanders Brine Shrimp) which included Schizochytrium sp. (SZ; Docosa Gold), Spirulina platensis (Sp) [=Arthrospira platensis] and Hematococcus pluvialis (H) either alone or in combination with partial rations of living microalgae. Growth of mussels fed 1/4 ration of living algae plus a mixed supplement of 50/50% w/w SZ:Sp grew significantly (SNK; P<0.05) faster than mussels fed a full live algal ration. Furthermore, growth (both wet weight and dry organic weight increase) of mussels fed diets composed of either 70/30% w/w SZ: ground Sp (GSp; <20 μm) or 25/50/25% w/w H:SZ:GSp was significantly (PLSD; P<0.05) greater than that of mussels fed an equal ration (by dry weight) of living Tahitian Isochrysis galbana.     

3种微藻对海水集约化对虾养殖尾水氮磷的去除效果

基于对虾生物絮团集约化养殖尾水含有高浓度硝态氮和磷酸盐的特征，比较分析钝顶螺旋藻（Spirulina platensis）、牟氏角毛藻（Chaetoceros muelleri）、盐藻（Dunaliella sp.）3种微藻在配制尾水中的存活生长状况及其对无机氮磷的去除效果，以期筛选出适宜的微藻用于后续尾水净化技术。采用显微镜计数法测定藻细胞密度，国标法测定总无机氮、氨氮、硝态氮、亚硝态氮和磷酸盐的含量。结果显示，钝顶螺旋藻在实验前后的藻细胞密度变化不大（P>0.05），约为3.32×106 个/mL和5.88×106 个/mL；牟氏角毛藻和盐藻细胞密度有明显增加（P<0.05），分别从初始的4.00×104 个/mL和2.50×105 个/mL升高至实验结束时的1.66×106 个/mL和1.06×107 个/mL。经过16 d实验，钝顶螺旋藻组对硝态氮和总无机氮去除率分别为79.60%和46.06%，显著高于其他各组（P<0.05），第8天时对磷酸盐的去除率可高达98.55%；牟氏角毛藻组16 d的磷酸盐去除率为98.25%，显著高于其他各组（P<0.05）。研究表明，3种微藻均可在对虾养殖尾水环境中存活，且对尾水氮磷具有较好的净化效果。     

凡纳滨对虾(Litopenaeus vannamei)工厂化养殖系统微藻的群落特征分析

2015年6-9月期间,对青岛市宝荣水产科技有限公司凡纳滨对虾(Litopenaeus vannamei)工厂化养殖车间的6个实验池进行采样调查,分析了水体中微藻的种类组成、丰度、多样性和优势种演替特征,并结合养殖情况进行了讨论.共检出微藻5门28属49种(其中优势种14种),丰度范围为5.2×105-9.4×108 cell/L,生物量范围为1.23-208.00 mg/L,多样性指数范围为0.42-2.44.多样性指数低于0.9时,生态系统稳定性差,对虾易发病.不同养殖阶段微藻优势种种类不同,前期主要是绿藻门(Chlorophyta)、硅藻门(Bacillarionphyta)和部分甲藻门(Pyrrophyta)的种类,中、后期以蓝藻门(Cyanophyta)的微囊藻(Microcystis sp.)和颤藻(Oscillatoria sp.)为主.对虾养殖密度显著影响微藻优势种演替,300 ind/m2养殖密度(A1池)藻相稳定且以绿藻和硅藻为优势种,对虾生长良好;400-500 ind/m2养殖密度(B1、C1和C2池)颤藻在中、后期演替成为绝对优势种,对虾易发病死亡.本研究为优化对虾工厂化养殖环境、指导养殖生产提供参考.     

Protein, carbohydrate, lipid and chlorophyll a content in Isochrysis aff. galbana (clone T-Iso) cultured with a low cost alternative to the f/2 medium

Growth, proximate composition and chlorophyll a content was estimated in static cultures of Isochrysis aff. galbana (clone T-Iso) to compare the quality and quantity of the biomass produced with an agricultural fertilizer medium versus f/2 medium. Culture of microalgae was done with controlled temperature, air flux of 4.5 l min⁻¹, salinity of 33‰ and an irradiance of 110±3 μmol m⁻² s⁻¹. Daily samples from the cultures were taken to estimate cellular density, proximate composition and chlorophyll a content during 7 days. Mean cellular density from samples with f/2 medium and with agricultural fertilizer were similar (no significance at P≤0.05), and mean growth rates of 0.62 and 0.61 doubling per day, respectively. Maximum similar values of protein content of 7.3 and 7.6 pg per cell were estimated on the 5th day in f/2 and agricultural fertilizer medium, respectively. Carbohydrate and lipid content decreased during the first 5 days corresponding to the exponential growth in both experiments. Higher content of carbohydrate (no significance at P≤0.05) was found in both media during the first 2 days, after 4 days the carbohydrates reduced three times theirs concentrations. Generally lipid contents in the cultures with agricultural fertilizer were significantly higher (P≤0.05) than f/2 medium. The chlorophyll a content increased exponentially during the culture, and its concentration was similar in both experiments. In conclusion, the cellular density and the biochemical constituents were similar when using either media. This fertilizer can be used in the production of I. galbana (T-Iso) employed in aquaculture.     

日本对虾仔虾培育试验

用无节幼体培育仔虾,采用的对虾幼体饵料有中肋骨条藻、卤虫无节幼体及台湾产的人工配合饵料,并注意饵料的配合使用及消毒工作;为维持水质稳定,整个培育期间不换水,加大布气石的密度（３．６个／ｍ２）。在３００ｍ３水体中,一茬育出仔虾２２１８万尾,成活率为６６．２％,取得了良好的经济效益。     

Feeding early larval stages of fire shrimp Lysmata debelius (Caridea, Hippolytidae)

An important constraint to the commercial rearing of the marine ornamental shrimp Lysmata debelius is high larval mortality during early stages due to inappropriate procedures of larval collection and not feeding a live prey before one day elapsed after hatching. This incorrect feeding practice is commonly adopted in larval rearing of L. debelius and other ornamental marine shrimps because it is wrongly assumed that reserves of the newly hatched are enough for the first 24 h of life. Present work demonstrates that captive newly hatched L. debelius larvae ingest microalgae within minutes after hatching. When fed solely with Artemia nauplii, they have acceptable survival rates with stocking densities at or below 50 larval L^–1; but when nauplii are combined with microalgae, survival is further improved to zoea 2 as initial mortality is reduced, and higher stocking densities are supported (up to 75 larvae L^–1). The microalgae used were Rhinomonas reticulata, Skeletonema costata and Tetraselmis chuii. Higher survival through metamorphosis to zoea 2 was always observed for groups fed combinations of microalgae including Tetraselmis chuii. It is recommended that, larval collection methods ensure that larvae are fed microalgae within 2–3 h of release.     

Effects of silage preservation on astaxanthin forms and fatty acid profiles of processed shrimp (Pandalus borealis) waste

Acid silage of shellfish processing waste has been reported to be a good and economical technique to protect these biomasses from bacterial decomposition. Shrimp (Pandalus borealis) by-products contain some value-added nutrients for the aquaculture industry such as carotenoid pigments (mainly astaxanthin) and n − 3 polyunsaturated fatty acids. The aim of this work was to determine the effect of ensiling shrimp waste during a long period of time (more than 3 months) on some unstable components such as the astaxanthin forms (free, mono- and diesterified) and polyunsaturated fatty acids. Comparisons of astaxanthin forms and the fatty acid profiles were performed on defrosted shrimp waste and on a 14-week-old shrimp waste silage. No significant difference (P ≥ 0.05) in the total lipids extracted from the two forms of shrimp waste was observed. Nevertheless, a small quantity of the red pigment, presumably an astaxanthin portion, was observed to stay firmly bound to the shrimp carapace after the solvent extraction in shrimp waste compared to the full recovery obtained in ensiled shrimp waste. This may explain the significantly (P < 0.05) higher concentration of total astaxanthin (4.57 vs 3.99 mg/g) found in the crude oil extracted from shrimp waste silage. Higher percentages (P < 0.01) of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) were found in esterified astaxanthin from shrimp waste silage (43.9% and 45.5%) in comparison with their shrimp waste (24.7% and 20.3%) counterparts. This suggests that EPA and DHA are the principal fatty acids esterified with the portion of astaxanthin linked to chitin in the shrimp carapace. The utilization of shrimp waste silage as a pigmenting component of salmonid feeds is also discussed.     

Aurantiochytrium sp. meal can replace fish oil in practical diets for the juvenile Pacific white shrimp

This study evaluated the use of microalgae (Aurantiochytrium sp.) meal as a substitute for fish oil in the diet of juvenile Pacific white shrimp (Litopenaeus vannamei) reared in a clear‐water system. Dietary treatments at five replacement levels (0%, 25%, 50%, 75%, 100%) were performed in triplicate. After 46 days, only a slight difference in shrimp final weight was observed among treatments (0.61 g). An increase in final weight was observed with replacement of up to 50% fish oil for microalgae meal, while the optimal percentage of replacement estimated was 44.7%. Feed conversion rate (FCR) of animals fed a diet of Aurantiochytrium sp.meal to replace up to 50% fish oil decreased, and the optimal percentage of replacement estimated was 49.3%. The fatty acids profile in shrimp muscle tissue demonstrated an increase in docosahexaenoic acid (DHA) from 10.03% to 14.28% with increased replacement of fish oil by microalgae meal in the diet. Therefore, the partial replacement of fish oil by microalgae meal resulted in improved shrimp growth and FCR, and total replacement of dietary fish oil had no negative effects on these parameters. In addition, inclusion of microalgae meal raises the level of DHA in shrimp muscle.     

Ontogenetic variability of chill tolerance in larval Artemia salina: II. Dual cryoprotectants

Chill tolerance was determined for various larval stages of Artemia salina in solutions with and without dual cryoprotectants (glycerol and sucrose) and in saline solutions of corresponding osmolalities. Synchronous batch cultures of larvae were removed from a donor culture at post-hatch intervals (0, 3, 6, 12 and 24 h) and transferred immediately to solutions at -1 ± 0.2°C for 10-, 30- and 90-min intervals. Survival of control groups (cryoprotectant-free sea water, salinity 40 ppt) varied with age. In these groups, 0 h post-hatch larval survival decreased linearly with exposure time to 27.1%. Older stages, however, were insensitive to prolonged -1°C exposure after the initial 10-min period. The addition of combined glycerol-sucrose cryoprotectants to the chilled solutions enhanced survival patterns, with 0h post-hatch nauplii showing the greatest enhancement and 3-, 6- and 12-h nauplii the least. Maximum survival enhancement exceeded 250% of controls. A 4 M glycerol concentration provided more cryoprotection than 1, 2 or 3M. No enhancement was afforded any age groups maintained under hyper-saline conditions.     

浮游微藻生态调控技术在对虾养殖应用中的研究进展

微藻技术是一项有机结合藻类生物学、藻类生态学及水生生态系统学的新兴生物技术,它以水体中的浮游微藻为对象,充分运用微藻的生理、生态特点,进而用以调控和优化水体生态环境。文章就当前在水产养殖领域中的微藻生态调控技术的相关研究进行了系统的综述,并对其在对虾养殖中的应用进行了展望。     

Bioprospection of Microalgae and Cyanobacteria as Biocontrol Agents Against Vibrio campbellii and Their Use in White Shrimp Litopenaeus vannamei Culture

Vibrio spp. are the most common and harmful shrimp pathogenic bacteria; however, microalgae and cyanobacteria have the ability to produce antimicrobial substances against these species. In this study, the organic and aqueous extracts of 28 species of marine microalgae and cyanobacteria were screened against Vibrio campbellii M1. Two of these phytoplankton species with antibacterial activity in aqueous extracts (Dunaliella tertiolecta and Skeletonema costatum) and nontoxic to brine shrimp Artemia franciscana nauplii were used to evaluate their anti‐Vibrio effect when used as green‐water cultures in Vibrio‐challenged white shrimp Litopenaeus vannamei cultures. No differences in mortality of juvenile L. vannamei were observed between treatments tested, suggesting that the pathogenicity of V. campbellii could be related to the growth stage of shrimp. The proximal composition of D. tertiolecta and S. costatum was in the recommended range for penaeid shrimp nutrition, allowing shrimp supplemented with these microalgae to have significantly greater total length and weight than control shrimp. Shrimp supplemented with S. costatum presented the highest values of organic mass (11.48 mg/organism) and growth rate (0.31 mg/d) in comparison to D. tertiolecta. These results indicate that microalgae are not only capable of producing antibacterial compounds against Vibrio but can also help shrimp nutrition.