The effects of light and temperature on the photosynthesis of the Asparagopsis armata tetrasporophyte (Falkenbergia rufolanosa), cultivated in tanks

The integrated aquaculture of the tetrasporophyte of Asparagopsis armata Harvey (Falkenbergia rufolanosa) using fish farm effluents may be viable due to the species high capacity of removing nutrients and its content of halogenated organic compounds with applications on the pharmaceutical and chemical industries. In order to optimize the integrated aquaculture of F. rufolanosa, we followed the daily variation of the potential quantum yield (F_v/F_m) of PSII on plants cultivated at different biomass densities and different total ammonia nitrogen (TAN) fluxes to check if they are photoinhibited at any time of the day. Moreover, the photoinhibition under continuous exposure to highly saturating irradiance and its potential for subsequent recovery in the shade was assessed. The potential for year round cultivation was evaluated by measuring rates of O₂ evolution of plants acclimated at temperatures ranging from 15 to 29 °C, the temperature range of a fish farm effluent in southern Portugal where an integrated aquaculture system of F. rufolanosa was constructed.Photoinhibition does not seem to be a major constrain for the integrated aquaculture of F. rufolanosa. Only when cultivated at a very low density of 1.5 g fresh weight (FW) l^− 1 that there was a midday decrease in maximal quantum yield (F_v/F_m). At densities higher than 4 g FW l^− 1, no photoinhibition was observed. When exposed to full solar irradiance for 1 h, F. rufolanosa showed a 33% decrease in F_v/F_m, recovering to 86% of the initial value after 2 h in the shade. A midday decline of the F. rufolanosa F_v/F_m was also observed under the lowest TAN flux tested (∼6 μM h^− 1), suggesting that this fast and easy measurement of fluorescence may be used as a convenient diagnostic tool to detect nutrient-starved unbalance conditions of the cultures. Maximum net photosynthesis peaked at 15 °C with 9.7 mg O₂ g dry weight (DW)^− 1 h^− 1 and remained high until 24 °C. At 29 °C, the net oxygen production was significantly reduced due to a dramatic increase of respiration, suggesting this to be the species' lethal temperature threshold.Results indicate that F. rufolanosa has a considerable photosynthetic plasticity and confirm it as a good candidate for integrated aquaculture at temperatures up to 24° C and cultivation densities of at least 5 g FW l^− 1. When cultivated at these densities, light does not penetrate below the first few centimetres of the surface zone. Plants circulate within the tanks, spending around 10% of the time in the first few centimetres where they are able to use efficiently the saturating light levels without damaging their photosynthetic apparatus.     

Exploring Northeast American and Asian species of Porphyra for use in an integrated finfish-algal aquaculture system

Many aquaculture industries generate a nutrient-rich waste stream that can lead to eutrophication of coastal waters. To address this environmental issue, the bioremediation potential of several native Northeast American species of Porphyra was assessed and compared to the well-known Asian species. Porphyra thalli were cultured over 4 weeks at 15 °C at a stocking density of 0.4 g FW L^− 1. At 3- to 4-day intervals nutrient uptake, tissue N accumulation and phycobiliprotein concentration (PBP) were determined as functions of nitrogen (N) concentration (25-300 μM) and N source (nitrate vs. ammonium). Growth rates were measured weekly. Growth and tissue N reached maximal levels at inorganic N concentrations of 150-300 μM. Maximum growth rates ranged from 10% to 25% day^− 1, although induction of archeospores reduced average growth rates in many cases. No evidence of ammonium toxicity (reductions in growth rate) was observed; in fact, similar values were found with both N sources. Ammonium generally yielded higher PBP and tissue N contents than nitrate. Porphyra amplissima presented the highest growth rate, followed by the Asian Porphyra yezoensis. Under the experimental conditions, Porphyra spp. removed 70-100% of N within 3-4 days at N concentrations up to 150 μM, but was less efficient in removing inorganic phosphorus (35-91% removal). The highest tissue N and PBP concentrations were found at 150-300 μM of N, with N values close to 7% DW. Overall, Porphyra appears to be an excellent choice for bioremediation of moderately eutrophic effluents, with the added benefit that tissue may be harvested for sale.     

The influence of stocking density, light and temperature on the growth, production and nutrient removal capacity of Porphyra dioica (Bangiales, Rhodophyta)

The optimal conditions for growth of Porphyra dioica gametophytes were investigated in the laboratory, focusing on bioremediation potential. Porphyra dioica is one of the most common Porphyra species along the northern coast of Portugal and can be found year-round. The influence of stocking density and photon flux density (PFD) on the growth, production and nutrient removal was tested. Maximum growth rates, up to 33% per day, were recorded with 0.1 g fw l^− 1 at 150 and 250 μmol photons m^− 2 s^− 1. Growth rate decreased significantly with increasing stocking density. Productivity (g fw week^− 1) had an inverse trend, with more production at the higher stocking densities. At 150 μmol m^− 2 s^− 1 and with 1.5 g fw l^− 1, 1.4 g fw week^− 1 were produced. At this PFD, there was no significant difference in production between 0.6 to 1.5 g fw l^− 1. Nitrogen (N) content of the seaweeds decreased with increasing stocking densities and PFDs. The maximum N removal was recorded at 150 μmol m^− 2 s^− 1, with 1.5 g fw l^− 1 stocking density (1.67 mg N day^− 1). However, the N removed by thalli at 50 μmol photons m^− 2 s^− 1 was statistically equal to that at 150 and 250 μmol photons m^− 2 s^− 1, at a stocking density of 1.0 g fw l^− 1. The influence of temperature and photoperiod on growth and reproduction was also assessed. Growth rates of P. dioica were significantly affected by temperature and photoperiod. In this experiment (with 0.3 g fw l^− 1 stocking density), the highest growth rate, 27.5% fw day^− 1, was recorded at 15 °C and 16 : 8¯, L : D¯. Male thalli started to release spermatia 21 days after the beginning of the experiment, in temperatures from 10 to 20 °C and with 10, 12 and 16 h of day length. Unfertilized female-like thalli were observed at 10 to 20 °C, under all photoperiods tested. Growth of these thalli declined after 4 weeks. By then, formation of young bladelets in the basal portion of these thalli was observed. After 7 weeks all biomass produced was solely due to these vegetatively propagated young thalli, growing 22.4% to 26.1% day^− 1. The results of this study showed that P. dioica appears to be a candidate as a nutrient scrubber in integrated aquaculture systems.     

Enhanced production of the sea urchin Paracentrotus lividus in integrated open-water cultivation with Atlantic salmon Salmo salar

Survivorship and performance was investigated for two size classes of the sea urchin Paracentrotus lividus grown adjacent to open-water Atlantic salmon (Salmo salar) cultivation on the north-west coast of Scotland. Juvenile P. lividus were maintained for 12 months at 0 m, 50 m and 2.5 km from the mariculture activity. The sea urchins at the 0 m station showed higher survivorship than at the 50 m and 2.5 km stations and had significantly greater test diameter than at the 50 m station. Only urchins at 0 m developed gonads and, although small, these were of acceptable or excellent colouration in terms of their marketability. Adult P. lividus were maintained for 3 months at 0 m and 15 m from the mariculture activity, either with or without additional macroalgae Laminaria spp. Sea urchins at the 0 m station and fed additional macroalgae showed higher gonadal growth than sea urchins at the 15 m station held without additional food. Gonad colouration in the adult urchins, irrespective of the treatment, was acceptable or excellent. The 0 m station also received significantly greater quantities of particulate organic matter (POM) than the other stations in both the juvenile and adult experiments. At the 0 m station, the presence of the fatty acid 22:1n-11 and elevated levels of the long chain n-3 fatty acid DHA 22:6n-3 in the gonadal tissue of the urchins confirmed the consumption of fish farm derived POM and highlighted the potential human health benefits from consuming sea urchin roe.The results show that P. lividus can thrive in the salmon culture environment and suggests that the integration of P. lividus with Atlantic salmon can provide a viable means to culture this species, even at this northerly latitude. Salmon-sea urchin co-culture would enable fish farms to diversify into producing a second highly valuable product and would reduce the increasing worldwide pressure of sea urchin fisheries on wild stocks.     

Ammonia toxicity and its effect on the growth of the South African abalone Haliotis midae Linnaeus

The current study investigated acute toxicity to ammonia of the South African abalone, Haliotis midae, from three size classes relevant to mariculture operations, and the chronic impact of sub-lethal ammonia levels on growth of juvenile abalone.Results showed that tolerance to ammonia (at pH 7.8 and T_a = 15 °C) increases with body size (i.e. age) as indicated by 36 h LC₅₀ values: juvenile abalone (1-2.5 cm shell length) had the lowest LC₅₀ of 9.8 μg l^− 1 FAN, whereas LC₅₀ was 12.9 μg l^− 1 FAN in “cocktail”-size abalone (5-8 cm shell length). The highest LC₅₀ of 16.4 μg l^− 1 FAN was observed in “brood stock”-size animals (10-15 cm). When “cocktail”-size abalone were allowed to acclimatize to sub-lethal ammonia levels for 48 h, their ammonia tolerance increased compared with non-acclimatized abalone of the same size: LC₅₀ was 2.0 μg l^− 1 FAN higher at 14.8 μg l^− 1 FAN.Growth of juvenile abalone (1-2.5 cm shell length) during chronic exposure to sub-lethal FAN levels is inhibited: specific growth rate (SGR) is significantly reduced by 58.7% to 0.10 ± 0.03% d^− 1 (weight) compared with 0.24 ± 0.06% d^− 1 of abalone of a control group (no ammonia).The results demonstrate the negative effects of ammonia not only on survival but also on growth of farmed abalone, both impair profitability of the farming operation. The information from the present study will be helpful in determining water quality requirements in South African abalone farms.     

Comparison of diagnosis techniques for the protozoan parasite Bonamia ostreae in flat oyster Ostrea edulis

In recent years, the cultured production of the flat oyster Ostrea edulis has suffered a dramatic decrease in Europe partially attributed to the protozoan parasite Bonamia ostreae, the causative agent of the Bonamiosis. In this paper the results of a PCR assay for the diagnosis of B. ostreae were compared with those obtained using two classical methods of diagnosis recommended by the OIE (Office International des Epizooties) and the European Union, histology and cytology. The same samples were analyzed by two different laboratories, showing that the results obtained with the PCR method have high sensitivity and good correlation between laboratories. This method is cheaper and faster than histopathology and cytology, with no need of specifically trained personnel to perform the diagnoses. It is appropriate for fast screening of stocks of great numbers of oysters.     

Evaluation of three seaweeds Gracilaria bursa-pastoris, Ulva rigida and Gracilaria cornea as dietary ingredients in European sea bass (Dicentrarchus labrax) juveniles

The aim of this study was to evaluate the inclusion of three seaweeds Gracilaria bursa-pastoris (GP), Ulva rigida (UR) and Gracilaria cornea (GC) as dietary ingredients on the performance, nutrient utilisation and body composition of European sea bass juveniles. Six experimental diets were formulated to replace 5% (GP-5, UR-5, and GC-5 Diets) and 10% (GP-10, UR-10 and GC-10 Diets) fish protein hydrolysate (CPSP) by each of the three seaweeds. A control diet was used, without inclusion of any seaweed. Diets were fed to duplicate groups of 25 European sea bass (Dicentrarchus labrax) juveniles (IBW = 4.7 g) for 10 weeks. Growth performance was only significantly reduced (P < 0.05) in fish fed the GC-10 diet, whereas the feed conversion ratio increased significantly in those fish. The apparent digestibility coefficients of dry matter and lipid were significantly lower in fish fed diet GC-10 relative to those fed the control diet. Carcass composition was similar among treatments, although fish fed GC-10 exhibited significantly higher ash content.The results obtained in this study suggest that the inclusion of G. bursa-pastoris (GP) and U. rigida (UR), up to 10%, can be considered as very interesting ingredients in diets for sea bass juveniles, as no negative consequences on growth performance, nutrient utilization or body composition were observed. On the other hand, the inclusion of G. cornea (GC) should be limited to 5% of the diet.     

Investigating fragment size for culturing reef-building corals (Porites lobata and P. compressa) in ex situ nurseries

Culturing small propagules of coral has the potential for high yield with low environmental impact, provided that mortality is low. This study investigated the size-specific mortality and growth of minute fragments (as small as 5 polyps) of two of the most abundant reef-building corals in Hawaii (Porites lobata and P. compressa). Two ex situ nursery systems differing in cost, design complexity, and labor intensity were compared. The first nursery experiment lasted for four months in a large tank with high water motion/surge and extensive manual cleaning to remove competitive algae. The corals were then transferred to a simple low-flow tank containing sea urchins (Tripneustes gratilla) and reduced cleaning, where they were grown for six more months (Nursery II). ‘Nursery I’ resulted in 92% of P. lobata and 73% of P. compressa fragments surviving and nearly doubling in area, in spite of a brief infestation by a nudibranch (Phestilla sibogae) that primarily fed on larger P. compressa fragments. There was a significant positive relationship between fragment size and growth rate, and survivorship was significantly different between donor colonies (genets), but there was no evidence of size-specific mortality. ‘Nursery II’ on the other hand had clear size-specific mortality and higher urchin damage for smaller fragments, resulting in moderate survivorship (78% for P. lobata and 76% for P. compressa), and only a slight increase in the total area covered by coral tissue. Fragments larger than 3 cm² were undamaged and had the highest survival and growth rates. This study illustrates how size-specific mortality can be reduced by ex situ nursery conditions.     

Experimental integrated aquaculture of fish and red seaweeds in Northern Portugal

Three potentially valuable red seaweeds, Chondrus crispus Stackhouse, Gracilaria bursa pastoris (S.G. Gmelin) P.C. Silva and Palmaria palmata (L.) O. Kuntze, collected in northern Portugal, were cultivated using the nutrient-rich effluents from a local turbot (Scophthalmus maximus Linnaeus) and sea bass (Dicentrarchus labrax Linnaeus) farm. The algae were cultivated in a two level cascade system. Several arrangements of the cascade system, stocking densities (3, 5, 7 and 8 kg m^− 2) and water fluxes (140 and 325 l h^− 1) were tested to optimize biomass yield and nitrogen uptake rate and efficiency. The yield and the total ammonium nitrogen (TAN) uptake of the three species were highly seasonal. Palmaria could not survive culture conditions during the summer when water temperature was above 21 °C. In the spring, Palmaria had an average yield of 40.2 (± 12.80) g DW m^− 2 day^− 1 and a nitrogen uptake efficiency (NUE) of 41.0% (± 17.26%). NUE expresses, in percentage, the average reduction in TAN concentration between the inflows and the outflows of the tanks. Chondrus performed better in summer with an average yield of 37.0 (± 11.10) g DW m^− 2 day^− 1 and removing 41.3% (± 17.32%) of nitrogen. Gracilaria grew year round, but also performed better during spring/summer, producing an average of 29.1 (± 2.90) g DW m^− 2 day^− 1, and only 7.3 (± 5.08) g DW m^− 2 day^− 1 during autumn. Yield of C. crispus did not differ significantly when grown at two different stocking densities (5 kg m^− 2 and 8 kg m^− 2). On the other hand, Gracilaria had significantly higher yields at 5 than at 7 kg m^− 2. Better NUE, on average 76.7% (± 22.13%), was also obtained with 5 kg m^− 2 stocking density and only 63.8% (± 24.62%) with 7 kg m^− 2. The yield of Gracilaria increased significantly with the increase of water flux from 140 to 325 l h^− 1 and more nitrogen was removed from the water. However, NUE decreased from 48.4% to 33.4% at 140 and 325 l h^− 1, respectively. Biofiltration was highly improved by a cascade system with a NUE as high as 83.5%.     

半滑舌鳎成鱼开放流水与循环水养殖模式下生长及肌肉营养成分差异研究

为半滑舌鳎的集约化养成提供基础数据,设定开放流水和循环水两种养殖模式,以处于快速生长期的大规格鱼种(0.640 ±0.063) kg为研究对象,检测半滑舌鳎的生长及肉质相关指标,实验共进行7个月,结果显示:①半滑舌鳎在两种养殖模式下的成活率分别为79.89％和90.38％,月增重率分别是0.154 7和0.1109 kg,肥满度分别是0.920和0.838,生长激素分泌量分别是2.812和2.706 μg/L.循环水养殖模式组在成活率、增重率、肥满度和生长激素分泌量上分别高于开放流水养殖模式组13.13％、39.5％、9.8％和3.92％.②两种养殖模式,开放流水养殖模式组在粗蛋白、氨基酸、PUFA上更占优势,但在粗脂肪、MUFA上循环水养殖模式明显占优势,分别高于开放流水养殖模式组41.732％和16.912％.实验表明,循环水养殖模式优越的水质环境更适合半滑舌鳎的育肥.本实验说明,半滑舌鳎在循环水养殖模式下具有很好的适应性,其生长处于较好的状态,营养品质上也能够得到保证.同时,也说明循环水养殖模式可以极大地发掘半滑舌鳎的养殖潜力,是一种适合半滑舌鳎集约化养殖的具有优势的养殖模式.     

Effects of Echerichia coli lipopolysaccharides and dissolved ammonia on immune response in southern white shrimp Litopenaeus schmitti

The success of penaeid shrimp aquaculture has been limited by endemic and epidemic infectious diseases around the world and shrimp defense mechanisms are a priority for control, prevention, and diagnosis. The lethal dose (LD₅₀) of Escherichia coli lipopolysaccharides was calculated and pathogen injection and dissolved ammonia concentration on immune response were evaluated in southern white shrimp Litopenaeus schmitti. The lethal dose of lipopolysacharides was 3.78 mg kg^− 1 body weight. Injection caused changes in phenoloxidase activity in plasma and nitric oxide and total haemocyte counts within the first 24 h. High concentration of dissolved ammonia caused a decrease in haemocytes by 66% within the first 72 h, when compared to the control. Hemagglutination in plasma was not affected by injection or high concentrations of dissolved ammonia. The results showed that white shrimp recognized the lipopolysacharides and responded to this microbial elicitor, as indicated by a variety of immunological indicators and that increased dissolved ammonia affected the number of circulating haemocytes.     

Distribution of a fish pathogen Listonella anguillarum in the Japanese flounder Paralichthys olivaceus hatchery

The present study was undertaken to investigate the distribution of Listonella anguillarum in a Japanese flounder (Paralichthys olivaceus) hatchery. A total of 2704 isolates were obtained from the developing fish, live diets and artificial feeds of Japanese flounder and their rearing water, 439 of which were identified as L. anguillarum by the combining incubation on thiosulfate-citrate-bile salt-sucrose (TCBS) agar at 35 °C overnight with polymerase chain reaction (PCR) detection for the VAH1 hemolysin gene. L. anguillarum was detected in all seven rotifer samples, with densities of 2.5 × 10³ to 4.6 × 10⁶ colony forming units (CFU) g^− 1. Both the analyzed samples of Nannochloropsis oculata contained this bacterium at densities of 1.6 × 10⁴ to 1.4 × 10⁵ CFU g^− 1. L. anguillarum was detected in only one of four samples of Artemia nauplii with a density of 4.8 × 10⁵ CFU g^− 1 (35%) and it was not detected in the two analyzed artificial feed samples. L. anguillarum was detected in 11 of 18 specimens of larval and juvenile Japanese flounder at densities of 5.0 × 10¹ to 7.4 × 10⁵ CFU g^− 1, while it was not detected in the two analyzed egg specimens of Japanese flounder. These results indicate that L. anguillarum associated with the developing Japanese flounder is likely derived from rearing water and live diets such as rotifers. Further, it is strongly suggested that L. anguillarum is a transient bacterium of the intestinal microflora for the Japanese flounder but is a permanently indigenous one for the Japanese flounder hatcheries.     

副溶血弧菌对斑节对虾非特异性免疫酶活性的影响

为了研究副溶血弧菌对斑节对虾非特异性免疫酶活性的影响,分别对斑节对虾注射生理盐水和副溶血弧菌,于不同时间点测定肝胰腺和鳃中的总抗氧化能力(T-AOC)、酸性磷酸酶(ACP)、碱性磷酸酶(ALP)和溶菌酶(LSZ)的活性变化。结果显示,与对照组相比,感染副溶血弧菌后,肝胰腺和鳃中T-AOC活性分别于12和6 h达到最大值(P0.05),随后逐渐降低;肝胰腺中ACP活性于3 h显著升高,随后逐渐降低,并于24 h达到最小值(P0.05),而鳃中ACP活性于12 h达到最大值,随后逐渐降低,并于48 h显著低于对照组(P0.05);肝胰腺和鳃中ALP活性整体均呈现出先升高后下降的趋势,但未出现显著低于对照组的现象;肝胰腺和鳃中LSZ活性均于3 h显著升高至最大值(P0.05),随后逐渐恢复至初始水平。研究表明,副溶血弧菌感染对斑节对虾非特异性免疫酶活有显著影响,对其机体免疫防御系统有明显的破坏作用。     

合浦珠母贝表皮生长因子样(EGF-like)基因的克隆与表达分析

为探究表皮生长因子(epidermal growth factor,EGF)在合浦珠母贝幼体发育以及组织生长过程中的作用,实验通过RACE技术克隆了合浦珠母贝EGF-like基因并做了相应的表达分析。实验结果获得cDNA全长序列4 107 bp,命名为Pf-egf1,开放阅读框(ORF)702 bp,编码234个氨基酸,包含一个信号肽序列和一个跨膜结构域,功能结构域分析发现Pf-egf1具有一个EGF-like结构域,有6个半胱氨酸残基,由3个二硫键维持,是表皮生长因子家族及其相关蛋白的特征结构域,但其余序列与现有相关基因序列差异较大,推测可能是一个新的EGF-like基因。表达结果显示,Pf-egf1 mRNA在合浦珠母贝外套膜、闭壳肌、鳃、肝胰腺、珍珠囊、肠和性腺中均有表达,在肠中的表达显著高于其他组织;在合浦珠母贝幼体发育的担轮期、D型期、壳顶期、眼点期和变态期的表达呈现逐渐升高的趋势,并且其在变态期的表达量极显著地高于其他时期。上述结果表明,Pf-egf1基因可能在合浦珠母贝肠道修复和幼虫变态发育阶段起着重要作用,为进一步开展育珠与生长调控奠定了基础。     

一株引起坛紫菜绿斑病病原的分离鉴定及致病性研究

为了明确引起坛紫菜绿斑病的病原,对2012年发生在福建省莆田养殖海区坛紫菜叶状体绿斑病开展了病原的分离鉴定和致病性研究。利用2216E海水培养基平板,从患病藻体分离得到一株优势细菌X5;在实验室条件下将X5进行回接感染实验,在显微镜下观察到坛紫菜叶状体发生组织病理变化,出现绿斑病症状:感染初期出现黄绿色小斑点,病斑逐渐扩大出现空洞,最后导致紫菜叶片流失;用浓度为10⁴~10⁸ cfu/mL的X5感染坛紫菜时,在7 d内引起紫菜出现10%~100%病斑面积,这些结果表明X5为坛紫菜绿斑病的病原。利用生理生化和多基因序列分析方法对X5进行鉴定,结果显示该菌为革兰氏阴性菌,大小为0.6 μm×0.4 μm~1.0 μm×0.4 μm,无鞭毛,在温度4~42℃、盐度50~150、pH6~10范围内生长,最适生长温度为16℃,最适生长盐度为50,最适pH为7,对甲硝唑、林可霉素和青霉素耐药,对25种抗生素敏感,Biolog和API ID-32E细菌鉴定系统的结果显示X5为弧菌属细菌(Vibriosp.);基于多基因序列(16S rRNA,rpoA,recA,pyrH)构建的系统进化树分析显示,X5与V.casei、V.litoralis、V.rumoiensis聚为一支,进化距离为0.095~0.108,这些结果说明X5为弧菌属的一个新种或是V.casei、V.litoralis、V.rumoiensis之一的一个新亚种。本研究报道了弧菌X5能够引起坛紫菜绿斑病,为紫菜流行病学及疾病防治提供了理论支撑。     

Experimental evaluation of a composted seaweed extract as microalgal culture media

An organic culture media based on a composted liquid seaweed extract was developed and tested for growth of Chaetoceros muelleri. The extract was evaluated at two concentrations and then compared with two commercial media: Walne and agricultural fertilizer (AF). The concentrations of nitrogen and phosphorous differed between the two commercial media and the seaweed extract; however, growth of Chaetoceros muelleri was similar. The successful culture of Chaetoceros muelleri in composted liquid seaweed extract suggests the feasibility of and potential use for this natural, organic fertilizer in aquaculture. The seaweed extract is simple to obtain and inexpensive.     

Effects of dietary carotenoid on the gut and the gonad of the sea urchin Paracentrotus lividus

Gonad colour, determined by accumulated carotenoids, is an important marketability factor in sea urchin products. In this study, dietary carotenoids were fed to Paracentrotus lividus in prepared diets to test their effects on the sea urchin's gonad colour. All diets had the same basic content of protein, lipid, carbohydrate, ash and energy. Four diets were enriched with: (I) all-trans β-carotene; (II) astaxanthin; (III) zeaxanthin/lutein/β-carotene; and (IV) capsanthin/zeaxanthin/lutein/cryptoxanthin/β-carotene. A fifth diet had Dunaliella bardawil algal powder containing all-trans and 9-cis β-carotene added, and a sixth one was a control diet with no added carotenoids. Sea urchins were fed the control diet (no pigments) for 4 weeks, then the six experimental diets for 8 weeks. Carotenoid concentration in the gut and gonad was determined by tri-dimensional photo-diode array high performance liquid chromatography. Four qualitative colour categories were established: brown, pale yellow, medium-orange, and mango-orange.Gonad development and test diameter increased equally in all diet treatments. Carotenoid profile and concentration in the gut were higher than in the gonad or in the feed. Total carotenoid concentration was four- to ten-fold greater in the gut than in the gonad, and carotenoid profile in the gut was different from that of the diet. Gut and gonad total carotenoids, β-carotene, and echinenone concentration were not dependent on total dietary carotenoid concentration. Total carotenoid and β-carotene concentration in the gut, and total carotenoid and echinenone concentration in the gonad were significantly dependent on dietary β-carotene concentration. The percentage of gonads with acceptable colour was positively correlated with dietary and gut β-carotene concentration. Carotenoids not normally contained in the diet of P. lividus (astaxanthin, capsanthin and capsorbin), did not accumulate in the gonad. Echinenone was found in all gut and gonad samples. Over 90% of the gonads from P. lividus fed Dunaliella algal powder diet developed the optimal mango-orange colour and this diet led to the greatest echinenone concentration.     

Comparison of Spore Inoculated and Vegetative Propagated Cultivation Methods of <Emphasis Type="Italic">Gracilaria chilensis</Emphasis> in an Integrated Seaweed and Fish Cage Culture

In Chile the integration of Gracilaria chilensis with salmon culture has shown high potential. Seaweed integrated aquaculture is of great interest as it allows waste recycling within fed cage aquaculture. The development of economically feasible suspended methods of seaweed cultivation is therefore of high importance. Hence, production and performance of two suspended Gracilaria cultivation methods, spore inoculated ropes and ropes with twined field collected seaweed, were studied in open water. The production from spore-seeded ropes was comparable to that of twined ropes for the first month of culture. Thereafter, the twined ropes had a significantly higher productivity. Fish farm wastes had no significant fertilizing effect upon Gracilaria growth rate. In addition, spore-originated thalli and field collected thalli were compared under laboratory conditions and in suspended culture using the same cultivation method. Spore-originated thalli had a 50% lower growth rate than the field collected thalli under laboratory conditions; however, no significant differences were detected in the field. Also, the occurrence of spore coalescence growth enhancement was not significant on the spore-seeded ropes. It was concluded that spore-originated cultivation techniques could be of interest for an integrated open seawater aquaculture system due to the high levels of Gracilaria polymorphism. This would result in greater adaptability to environmental variations, and a continuous supply of restocking material.     

Histopathology and a real-time PCR assay for detection of Bonamia ostreae in Ostrea edulis cultured in western Canada

Bonamia ostreae is an intracellular haplosporidian parasite in European flat oysters Ostrea edulis that occurs on both coasts of the United States and causes significant mortality in Europe. Canada was considered free of B. ostreae until 2004, when it was first detected in O. edulis obtained for laboratory study from a western Canadian oyster farm. Bonamia ostreae was confirmed in O. edulis at the index farm in November 2004 using histopathology, conventional polymerase chain reaction (PCR) assays, restriction fragment length polymorphism (RFLP) analysis, and sequencing of the PCR product. Archived samples of European flat oysters obtained from the index farm between 1999 and 2004 (n = 343) were re-examined and all samples collected before 2003 (n = 306) were confirmed negative for B. ostreae by histopathology (n = 306) and PCR (n = 62). In archived samples from 2003, B. ostreae was detected in 3 of 37 O. edulis by histopathology (n = 2) and/or PCR (n = 2). Also, records indicate that B. ostreae was not detected in O. edulis (n = 348) from five other locations in western Canada between 1986 and 2000. To better understand the distribution and prevalence of B. ostreae in western Canada, 607 oysters from the index farm and 2 additional farms were sampled in the summer of 2005. All 3 farms had been stocked with O. edulis spat from the State of Washington, USA, where B. ostreae is endemic. Samples were analyzed by histopathology and a new real-time PCR that amplifies a 68-bp target DNA fragment. B. ostreae was detected in all three locations, with prevalence ranging from 0.5 to 11.1%. Diagnostic sensitivity of the real-time PCR method was consistently greater than histopathology. Also, preliminary evidence supports the conclusion that real-time PCR on paraffin sections is more sensitive than histopathology; B. ostreae DNA was confirmed in 4 oysters by real-time PCR on paraffin-embedded tissues (and independently confirmed on unfixed tissues) that was not detected by histopathology. As a result of these findings, O. edulis spat are no longer allowed to be imported from endemic areas into Canada.     

西伯利亚鲟海豚链球菌的分离鉴定及毒力基因检测

2013年8-9月,四川雅安汉源湖养殖的西伯利亚鲟发生一种以体表溃疡、内脏出血和心外膜囊肿为特征的传染病.为明确其病因,本研究从自然发病鱼的肝、脾和肾进行了病原菌的分离、人工感染、分离菌的表型特征和分子生物学特征的检测.结果从患病鱼体内分离到一株G+链状球菌(Ab130920),人工感染实验证实了其病原性,生理生化特性与海豚链球菌(ATCC29178)基本一致;16S rDNA序列(GenBank登录号:KJ162337)与GenBank中S.iniae 16S rDNA序列同源性最高,在以16S rDNA序列及其GenBank中同源性较高的相关序列构建的系统发育树上,分离菌与海豚链球菌聚为一支;同时,在基于S.iniae lctO基因的特异性PCR检测中,从分离株基因组DNA扩增出预期大小的870 bp条带,进而鉴定分离菌Ab130920为S.iniae.在对cpsD、simA、sagA、pdi和scpI等5种S.iniae毒力基因的多重PCR检测中,分离菌均扩增出相应大小的特异性片段,表明其为一毒力较强的菌株,与人工感染实验的高致病性结果相佐证;药物敏感性检测发现其对阿莫西林、强力霉素、氟苯尼考等抗菌药物敏感,但对新生霉素、利福平、氧氟沙星耐药.