轮虫、卤虫营养强化研究进展

轮虫和卤虫具有丰富的营养价值,是鱼类和甲壳类幼体培育的重要饵料。文章从轮虫和卤虫营养强化剂的选择、营养成分、强化方法和时间等层面概述了该领域的研究进展,可为开展轮虫、卤虫的营养强化以及在水产养殖中的应用提供参考。     

轮虫与卤虫在营养价值等方面的比较

近些年来,随着水产养殖业的发展,轮虫(rotifer)和卤虫(Artemia)由于适口性好、蛋白含量高、对水质污染小等优点,已被广泛地应用于鱼、虾、蟹类的苗种生产,成为不可替代的生物饵料。本文对轮虫和卤虫的生物学特性、研究历史、营养价值、培养方式和营养强化等方面进行了比较性的综述,以便对轮虫与卤虫有进一步的了解。     

不同饵料对锯缘青蟹幼体发育的影响

锯缘青蟹幼体培育实验表明在幼体培育过程中,单投轮虫、卤虫、人工饵料及藻类幼体的发育并不理想,均在某个变态阶段受阻.人工饵料和藻类在投喂的第四天已全部死亡.但适当添加人工饵料及藻类以几种饵料混合投喂方式可有效地提高幼体成活率.轮虫、丰年虫、裸腹蚤均是幼体良好的生物饲料.     

生物饵料的n－3 PUFA营养强化技术

生物饵料的ｎ－３　ＰＵＦＡ营养强化技术张利民（山东省海洋水产研究所，烟台２６４０００）在海水鱼虾育苗过程中，轮虫（Ｂｒａｃｈｉｏｎｕｓｐｌｉｃａｔｉｌｉｓ）和卤虫（Ａｒｔｅｍｉａｓａｌｉ－ｎａ）是重要的生物饵料，而不同的养殖种类对营养需求不一样，例如...     

细点石斑鱼工厂化人工繁育及养殖技术(中)

<正>5.饵料投喂策略初孵仔鱼→3天仔鱼(投放益生菌1毫克/升、投喂小球藻30万个/毫升、S型轮虫2个/毫升)→8天仔鱼营养强化(投喂S型或L型轮虫3个/毫升)→18天仔鱼(投喂卤虫或桡足类无节幼体0.5个/毫升)→28天稚鱼(停喂藻类和轮虫、投喂小卤虫或桡足类1个/毫升)→38天稚鱼(投喂卤虫或桡足类成体2个/毫升)→40天幼鱼(投喂颗粒饲料，投喂量为幼鱼体重的5%～8%)。     

芽孢杆菌微生态制剂对卤虫异养细菌的影响

芽孢杆菌是目前水产养殖中广泛应用的益生菌之一。目前已知的芽孢杆菌的主要作用包括改善养殖水体的理化和微生物指标、提高消化酶活力、增强免疫力,并对某些养殖对象的仔稚鱼的肠道菌群有调节作用。芽孢杆菌采用泼洒的施用方法需要大量并连续多次的使用才能取得显著效果,成本较高,且对生态环境具有潜在风险。近年来的研究表明,轮虫、卤虫、桡足类等饵料生物在投喂前用芽孢杆菌等益生菌菌液进行短时间、高浓度的浸泡处理,     

中湿度表面的海洋细菌对厚壳贻贝稚贝附着的影响

为研究自然微生物膜中海洋细菌对厚壳贻贝稚贝附着的影响,通过海洋贝类生物学、分子微生物生态学等手段调查附着基表面湿度、微生物膜的密度以及细菌种属系统发育与厚壳贻贝(Mytilus coruscus)稚贝附着关系。结果表明,所有测试海洋细菌形成微生物膜的最终密度随初始密度的增加而增加。所测海洋细菌均能不同程度地诱导厚壳贻贝稚贝的附着,其中Cobetia sp.3形成的微生物膜显示出最高诱导活性,其诱导的稚贝附着率为(70%±3%);Nautella sp.2、Pseudoalteromonas sp.9、Pseudoalteromonas sp.10、Bacillus sp.5和Pseudoalteromonas sp.11等5株细菌表现中等程度的诱导活性,其诱导的附着率范围为51%~60%。所有测试菌株所形成的微生物膜密度与稚贝附着均呈显著相关,尤其是Pseudoalteromonas sp.9和Cobetia sp.3诱导活性与附着率相关性极强,其相关性系数分别为0.774 1和0.723 3。系统发育分析结果表明,微生物膜密度和厚壳贻贝稚贝的附着率显著相关,然而海洋细菌诱导活性与细菌种属无关。因而,中湿度表面的海洋附着细菌对厚壳贻贝的附着有着明显的促进作用,本研究为后续开展厚壳贻贝稚贝的附着机制提供了理论依据。     

养殖刺参早期发育阶段体内可培养细菌的菌群特征及其与环境菌群相关性分析

采用传统细菌培养方法，对养殖刺参(Apostichopus japonicus)早期发育各阶段幼体体内及环境(投入饵料及培育用水)菌群的组成与结构展开研究，对分离的优势细菌进行分子鉴定，在此基础上，进行了刺参幼体体内菌群结构与环境菌群结构相关性分析。幼体各发育期的细菌培养结果显示，在幼体开口前的各发育时期(性腺、卵、受精卵、原肠胚)均无可培养细菌，在投饵以后，耳状幼体、樽形幼体体内可分离到可培养细菌，幼体发育到稚参以后，消化道可培养细菌总数急剧增加，并在4月龄时达到108 CFU/g数量级。在幼体体内可培养细菌中，弧菌(Vibrio)占比为2.2%~77.3%。对环境菌群的细菌培养结果显示，培育用水中细菌含量变化不显著，随着幼体发育期饵料的转变，不同时期饵料中细菌含量差异显著。整个养殖系统中共分离到65株优势细菌，16S rDNA鉴定结果显示，所分离的65株优势菌鉴定为14个属43种细菌。相关性分析结果显示，随着幼体的发育，生物饵料中的细菌对消化道中的菌群结构影响越来越大。本研究结果为解析刺参消化道菌群的形成过程和演替规律以及养殖用益生菌的筛选与应用奠定了基础。     

饵料生物营养强化剂及效用

水产育苗用饵料生物的微藻、轮虫、卤虫、枝角类等,经营养强化后,能提高水产种苗的健壮程度。本文就自动控制培养装置、冷冻浓缩微藻液,以及油液类、虫卵类、微囊粉末类及饵料生物代用品微粒饲料等营养强化剂及效用,进行分述。     

云斑尖塘鳢转饵期仔鱼生长和存活率的研究

在水温(30±1)℃条件下对人工孵化的云斑尖塘鳢(Oxyeleotris marmorata)仔鱼进行转饵期不同投喂策略实验,对仔鱼生长和成活情况进行了研究,设计5个投喂组:轮虫投喂组(A组);70%轮虫+30%卤虫投喂组(B组);50%轮虫+50%卤虫投喂组(C组);30%轮虫+70%卤虫投喂组(D组);卤虫(幼虫)投喂组(E组)。结果显示:云斑尖塘鳢仔鱼对卤虫的摄食强度随卤虫在饵料中比例的升高而增大,其对混合饵料中轮虫和卤虫的摄食强度的比值(Ir/Ia)随食物组成的变化发生相应的变化。以轮虫-卤虫不同比例的混合饵料投喂时,D组仔鱼的增长率、增重率最高,可存活率不足50%,高于E组,显著低于其它各组;C组仔鱼生长指标略低于D组,但存活率为(74.00±2.29)%,综合指标显著好于其它各组。当混合饵料中轮虫和卤虫各占50%时,10日龄开始投喂组仔鱼的全长(13.07±0.13)mm,特定生长率(10.25±0.37)%/d,存活率(82.14±0.72)%,综合生长指标最好。结果表明,云斑尖塘鳢仔鱼转饵期进行联合投喂的最佳饵料比例为50%轮虫+50%卤虫,最佳转饵时间为10日龄。     

Characterization of bacterial community composition in rotifer cultures under unexpected growth suppression

Unexpected growth suppression is common in rotifer mass culturing, and bacteria present in culture tanks are believed to be one of the main causes. We used a polymerase chain reaction (PCR)-amplified 16S ribosomal RNA (rRNA) gene finger-printing method to determine the bacterial community composition in water and that in association with rotifers present in tanks when rotifer growth was suppressed. Bacterial 16S rRNA genes representing Nautella sp. and Marinomonas sp. were identified as dominant in control tanks with good rotifer growth. In contrast, the bacterial community composition was more diverse at start-up in tanks where rotifer growth was suppressed. The community composition changed during rotifer cultivation, and bacterial sequences representative of Ruegeria sp. and Hyphomonas sp. were dominant when rotifer growth recovered. These findings indicate that the bacterial community composition varied according to the rotifer growth conditions and suggest that the initial microbial flora present in the culture water may cause rotifer growth suppression.     

Lactobacillus spp. bacteria as probiotics in gilthead sea bream (Sparus aurata, L.) larvae: Effects on growth performance and digestive enzyme activities

In this study, the influence of commercial probiotic, Lactobacillus spp., supplementation was investigated on growth parameters and digestive enzyme activities in gilthead sea bream, Sparus aurata, during larval development. All experiments were triplicated and designed in three different administrations of probiotic from 3 days after hatching (DAH) concurrently with starting of exogenous feeding. In the first group, probiotic was added to live food (rotifer and Artemia). In the second group, probiotic was supplemented directly to both live food and water. In the third group, probiotic was added directly to water. Also, no probiotic treatment was maintained in control group. Total bacterial counts among probiotic probiotic-supplemented groups were significantly different from total bacterial counts in controls in water and digestive tract of larvae (p < 0.05). The mean of total bacterial counts in control was approximately 4 × 10⁴-fold increased from the experimental groups in the sea water (p < 0.05). Besides, mean digestive enzyme activities of all probiotics treatment groups were significantly different (p < 0.05) with that of the control. Except probiotic water supplementation group, in all treatments, the specific activities of pancreatic and intestinal enzymes were significantly higher (p < 0.05) in larvae to which probiotic had been supplemented by live food and live food with water. Also, S. aurata larvae that had probiotic administered by live food with water demonstrated significant (p < 0.05) increases in both survival (13–105% higher) and specific growth rate (2–9% higher) as compared to controls. As a result, supplementation of probiotic to directly tank water could not significantly increase growth parameters and digestive enzyme activities and therefore, administration of probiotics by this method would not be effective in terms of husbandry parameters and nutritional condition.     

基于16SrDNA分析的浒苔外生细菌多样性研究

为研究浙江近海浒苔Ulva spp.(Enteromorpha spp.)外生细菌多样性,采用传统的形态学和16S rDNA测序分析的方法,从舟山朱家尖、宁海国华电厂和奉化南沙3个地区分离到可培养的浒苔外生细菌及其周围海水细菌65株。根据细菌菌落特征和革兰氏染色结果等将分离到的细菌分为26种表型。16S rDNA序列测序比对发现:菌株与不动杆菌属(Acinetobacter)、交替单胞菌属(Alteromonas)、弧菌属(Vibrio)、假交替单胞菌属(Pseudoalteromonas)、微小杆菌属(Exiguobacterium)、芽孢杆菌属(Bacillus)、赤细菌属(Erythrobacter)、微球菌属(Micrococcus)、假单胞菌属(Pseudomonas)、深海杆菌属(Idiomarina)、Phaeobacter、Roseivirgaj和Silicibacter等23个属相应菌株具有较高同源性。对不同地区浒苔外生细菌进行了Shannon-Wiener多样性指数分析。研究表明:(1)宁海国华电厂的浒苔外生细菌多样性最丰富,多样性指数为93.98%;(2)浒苔外生细菌与其生活地区密切相关,其群落组成具有地域差异,其优势类群也不尽相同,但均归属于变形菌门。     

PCR-DGGE指纹图谱技术分析发病龙须菜附生菌菌群组成

本研究以07-2品系龙须菜(Gracilaria lemaneiformis)为实验材料,提取自然条件下代表不同病变程度的健康、病间、白化藻体的附生菌DNA,经PCR扩增16S r DNA基因的V3可变区并进行变性梯度凝胶电泳(DGGE),分离V3片段,再对目的条带进行克隆鉴定。实验结果经数据化后计算龙须菜附生菌数目并比较附生菌菌群多样性,通过非度量多维尺度分析(n MDS)及多元梯度分析(RDA)来比较附生菌菌群组成相似性及优势菌群。实验结果表明,随着龙须菜病变程度的加深,附生菌菌落数及菌群多样性逐渐增加(P0.05)。n MDS分析结果表明,健康藻体与白化藻体在附生菌菌群组成上存在差异,病间藻体在附生菌菌群组成上与健康藻体更为接近。RDA分析结果表明,龙须菜健康藻体附生菌可能的优势菌群是海洋单胞菌属(Marinomonas),病间藻体附生菌可能的优势菌群是单胞菌属(Alteromonas)、红杆菌科(Rhodobacteraceae)、假交替单胞菌属(Pseudoalteromonas),白化藻体附生菌可能的优势菌未能鉴定到,而白化残留藻体附生菌鉴定到的优势菌有Maribacter、红杆菌科、玫瑰变色菌属(Roseovarius)。从上述结果可以看出,龙须菜健康藻体与发病藻体在附生菌菌群组成及优势菌上存在较大差异,而介于两者之间的藻体在菌群组成上更偏向于健康藻体。     

Pathogenic threats and probiotic use in larviculture of the scallop,Pecten maximus

During a series of experiments, bacteriological elements in scallop larval rearing were investigated: larvae susceptibility to pathogens as a function of their age, and the use of probiotic bacteria during larviculture. Younger larvae (d5 PF) were highly more susceptible to pathogenic‐challenge than their older siblings, which were challenged at an older age (d15 PF). A challenge with 10⁴ CFU mL^?1 of V. pectenicida killed 100% of d5 PF larvae 7 days following challenge, yet killed only 9% of d15 PF larvae 9 days following challenge. Use of the probiotics Phaeobacter gallaeciensis, Alteromonas macleodii 0444 and Neptunomonas sp. 0536, provided for larger larvae, a high yield of competent larvae and, perhaps more importantly, protection against pathogen‐challenge similar to levels achieved from antibiotic use. When challenged with V. pectenicida, d29 survivals were 20.3%, 85.1% and 75.0% respectively for control (no probiotic), antibiotic treated, and ‘probiotic mix’ administered larvae. Use of potential probiotic Pseudoalteromonas sp. D41 appeared to hinder scallop larvae. Future use of probiotics in scallop larval rearing would benefit from combined use of P. gallaeciensis, A. macleodii 0444 and Neptunomonas sp. 0536.     

Use of macroalgae supplemented with probiotics in the Haliotis rufescens (Swainson, 1822) culture in Northern Chile

In this study, we evaluate the use of macroalgae as vectors of probiotics bacteria into the digestive tract of abalone to improve their survival and growth. It is shown that when abalone Haliotis rufescens of different sizes were fed with a natural diet composed of fronds of the macroalga Macrocystis integrifolia supplemented with a mixture of Vibrio sp. C21‐UMA, Agarivorans albus F1‐UMA and Vibrio sp. F15‐UMA bacteria, there was a significant increase (P<0.05) in the average monthly growth rate and survival (%)in a period of 210 days, compared with the control without a probiotic supplement. The permanence of the probiotics in the digestive tract of the animals was monitored, and it was found that the number of culturable C21‐UMA and F1‐UMA bacteria decreased significantly (P<0.05) in recently weaned and adult abalone, and they almost disappeared completely on day 19 of the bioassay. However, the culturable Vibrio sp. F15‐UMA disappeared completely from the digestive tract on day 22 of the bioassay, and these were the bacteria that remained at the highest concentration compared with the other two bacterial strains over the experimental period. It is therefore shown that it is feasible to use a probiotic mixture to improve the profitability of the H. rufescens culture.     

Bacteria associated with early life stages of the great scallop, <Emphasis Type="BoldItalic">Pecten maximus</Emphasis>: impact on larval survival

A bacteriological study was carried out at a scallop (Pecten maximus) hatchery near Bergen, western Norway following a severe increase in mortality rates during the larval stages of the scallops. No larvae survived to settling, except for those in groups treated prophylactically with chloramphenicol. In order to identify pathogenic strains of bacteria, we performed a challenge test on 10- to 16-day-old larvae using isolated bacterial strains from the hatchery. Infection with six of these strains produced mortalities that were not statistically different from that resulting from infection with the known pathogen Vibrio pectenicida. However, about 5% of the strains tested in the challenge experiment produced higher motility rates than found in the unchallenged control group, indicating a possible probiotic effect. On the basis of 16S rDNA analysis on these strains, the phylogenetic tree indicated two groups of apparent pathogens: (1) one strain, LT13, grouped together with Alteromonas/Pseudoalteromonas; (2) a cluster of strains grouped together with Vibrio splendidus (LT06, LT21, LT73, PMV18 and PMV19). Strain LT13 was isolated from cultures of the microalga Chaetoceros calcitrans used for feed, while the other strains were isolated from larval cultures. Transmission electron microscopy showed intracellular bacteria that resembled bacteria in the groups Chlamydiaceae and Rickettsiaceae.     

培养基对微生物被膜形成和厚壳贻贝附着的影响

为探讨微生物被膜生物学特性、培养基以及大型生物附着三者之间的相互关系,本实验通过微生物分子生态学和海洋贝类生态学等方法调查了不同培养基对海洋细菌所形成微生物被膜的影响及其对厚壳贻贝幼虫附着的影响。结果显示,厚壳贻贝稚贝率与培养基类型和细菌初始密度显著相关,进一步分析表明,Zo Bell 2216E和Seawater Luriabertani(SWLB)条件下分别有6株和9株细菌形成的微生物被膜密度与稚贝率显著相关。扫描电镜结果显示,Staphylococcus sp.3在Zo Bell 2216E培养基条件下形成微生物被膜的细菌分布较为紧密,Pseudoalteromonas sp.8则在SWLB培养基条件下微生物被膜细菌分布较为紧密,且形态变为短杆状。SDS-PAGE结果显示,相比Zo Bell 2216E培养基,Staphylococcus sp.3在SWLB培养基条件下9条条带蛋白显著下降,其中2个条带完全消失;Pseudoalteromonas sp.8则在SWLB培养基条件下明显增加5条蛋白条带。研究表明,微生物被膜的形成受到培养基的影响,培养基的不同导致微生物被膜的形态结构、分布和蛋白有所差异,最终导致微生物被膜诱导厚壳贻贝幼虫附着变态的活性差异,本研究为后续开展厚壳贻贝附着的分子机制奠定良好基础。     

饲料添加益生菌对多病原阳性的凡纳滨对虾生长与存活的影响

采用假交替单胞菌(Pseudoalteromonas sp.) KL-3 2010和微小杆菌(Exiguobacterium sp.) KL-C2 2014作为益生菌，进行凡纳滨对虾(Litopenaeus vannamei)投喂实验，研究上述菌株对带毒对虾的生长与存活的影响。假交替单胞菌KL-3 2010对致急性肝胰腺坏死副溶血弧菌(Vibrio parahemolyticus) (VPAHPND 20130629002S01)有拮抗作用和胞外蛋白酶活性，微小杆菌KL-C2 2014有胞外蛋白酶活性。待试对虾经检测为白斑综合征病毒(WSSV)、致急性肝胰腺坏死病副溶血弧菌(VPAHPND)和虾肝肠胞虫(EHP)弱阳性。经过为期60 d的养殖实验，结果显示，与投喂普通颗粒饲料的对照组相比，投喂假交替单胞菌KL-3 2010的对虾存活率提高了213%±43% (P<0.01)；投喂微小杆菌KL-C2 2014的对虾平均生长率提高了105.5%±28.1% (P<0.05)；交替投喂2株菌的对虾存活率提高了184%±52% (P<0.05)，平均生长率提高了70.6%±32.8%。肠道可培养优势菌研究表明，2株益生菌的投喂显著影响了对虾肠道优势菌群的种类。本研究为带毒虾苗的养殖提供一种有效的病害防控和促生长的手段。     

Effects of feeding regime and probionts on the diverting microbial communities in rotifer Brachionus culture

Rotifer growth performance and microbial community changes associated with rotifer cultures were monitored while different feed types (Nannochloropsis oculata paste and the commercial yeast based feed CS-3000), different regimes (daily changes, changes per batch and no changes) and mixtures of three probionts (Phenylobacterium sp.; Gluconobacter sp. and Paracoccus denitrificans) were provided. It was shown that the dominant bacterial species in the cultures receiving either N. oculata or CS-3000 were different. However, in cultures receiving both feeds (either switching between feeds on a daily basis or on a batch basis), a high similarity in microbial community fingerprint was found. The presence of probionts was detected by the end of four batch culture cycles in spite of strong shifts of the bacterial community. By group discriminant analysis, it was found that Phenylobacterium sp. and Paracoccus sp. contributed positively to the CS-3000-fed group, while Gluconobacter sp. contributed positively to the N. oculata-fed group, although they did not appear as very dominant species.