Effects of Nitrate Concentration on Larval Development of the Giant River Prawn,Macrobrachium rosenbergii

Abstract

The effect of nitrate concentration on giant river prawn, Macrobrachium rosenbergii, larvae was investigated. Survival rate, weight gain, and larval development were evaluated for different concentrations of nitrate in three experiments. The experiments were divided in to two phases. In the first phase, larvae from stages I through VIII were analysed, while in the second phase larvae from stage VIII through post-larvae metamorphosis were analysed. Oxygen consumption was also determined for zoea I, II, and VIII exposed to 0, 700, and 1,000 mg/L of nitrate-N. No effect was observed for concentrations up to 180 mg/L NO₃-N (experiments I and II), and nitrate levels as 1,000 mg/L NO₃-N did not affect survival in the first phase of the third experiment. On the other hand, larval stage index (LSI) and weight gain decreased as nitrate-N concentration increased from 0 to 1,000 mg/L. In the second phase, survival and metamorphosis rate decreased as nitrate concentration increased, according to a linear model. The effect of nitrate levels on weight gain followed a curvilinear pattern. Larval respiration decreased in the water where nitrate was added, but only during stage II. The results demonstrated that nitrate presents extremely low toxicity for giant river prawn larvae, and data were related to the levels of nitrate that usually occur in larviculture systems also discussed. Therefore, nitrate is not a limiting factor for giant river prawn larviculture.     

Biochemical Changes in Freshwater Prawn Macrobrachium rosenbergii During Larval Development

Abstract.— Quantitative changes in the protein, lipid and carbohydrate were studied in the early larval stages of developing freshwater prawn Macrobrachium rosenbergii reared under fed and starved conditions to understand the relative importance of these nutrients in energy metabolism of the growing larvae. Larvae obtained from several females were stocked into three 250-L tanks at a density of 30 larvae/L. The feeding regime consisted of newly hatched Artemia nauplii. Protein was always the major organic constituent followed by lipids and then carbohydrates of both fed and starved larvae. Protein levels of both fed and starved larvae increased during development, suggesting an important role in morphogenesis. The decline of lipid during the larval growth that was more rapid for starved larvae, suggests a probable utilization of lipid as the major metabolic source of energy. Carbohydrates formed less than 5 and 2.4% of the larval dry weight of fed and starved larvae, respectively, suggesting their limited role in larval metabolism.     

人工海水培育罗氏沼虾幼体试验

按海盐∶MgSO4∶CaCl2∶KCl∶H3B3O3∶KBr∶EDTA∶生石灰为2000∶600∶72∶36∶24∶4∶1∶2的比例配制成人工海水,研究在该人工海水中培育罗氏沼虾幼体的可行性。结果表明,人工海水中的Mg2+、K+、Ca2+的比值为3∶1∶1,与天然海水接近,且水质无毒性作用;人工海水培育80万尾罗氏沼虾虾苗,获得68万尾幼体,成活率约为85%,蜕皮率为16.82%,特定生长率为25.34%/d,高于天然海水的培育水平。结果表明,自制人工海水可用于罗氏沼虾幼体培育。     

不同温度下2种沼虾磷酸酶活性变化的研究

用比色法测量4℃和-70℃下离体的罗氏沼虾和日本沼虾肝胰腺中的碱性磷酸酶、酸性磷酸酶酶活性在一周内的变化。试验结果表明,4℃条件下保存的日本沼虾与罗氏沼虾的离体组织中碱性磷酸酶和酸性磷酸酶酶活性有效测定下时间为3 d,-70℃下有效测定时间为2 d,对后续酶活性试验没有大的影响。     

高位池养殖罗氏沼虾试验报告

在黄河三角洲地区对高位海水对虾养殖池进行改造,利用低盐度水源进行罗氏沼虾养殖试验, 获得成功。经116天养殖,亩产罗氏沼虾50.1 kg,平均体长7.86cm,平均体重15.2 g,投入产出比1:2.4, 亩获纯利润1 205元。     

罗氏沼虾胚胎发育期主要消化酶和同工酶特性

采用生物化学方法分别测定了罗氏沼虾胚胎发育期5种消化酶:胃蛋白酶、类胰蛋白酶、淀粉酶、纤维素酶和脂肪酶的比活力,测定了6种同工酶:酯酶(EST)、苹果酸脱氢酶(MDH)、谷氨酸脱氢酶(GDH)、碱性磷酸酶(ALP)、乳酸脱氢酶(LDH)和淀粉酶(AMY)在成熟卵细胞和胚胎发育时期的表达特征。结果显示:5种消化酶的活力表现出两种变化趋势,即胃蛋白酶、类胰蛋白酶、淀粉酶和纤维素酶的酶活力呈现出“高—低—高”的变化趋势,而脂肪酶活力则呈现出下降的变化趋势。6种同工酶酶谱则随胚胎发育渐趋复杂,酶活性随之增强。研究表明,所测定的消化酶以及同工酶在罗氏沼虾受精卵期均已表现出酶活性,此阶段为卵源性的酶;原肠期是胚胎发育中一个较为重要的时期,胚胎内酶基因表达能力开始逐渐增强,开始合成一些酶类,为胚胎发育后期利用卵黄物质提供了保证。消化酶以及同工酶对卵黄物质的利用和胚胎发育的物质代谢起重要作用。     

青虾幼虾发育时期性腺发育组织学研究

采用组织切片及类固醇激素含量测定等方法,研究青虾Macrobrachium nipponense幼虾发育第1~31d的精巢、卵巢及促雄腺发育的起始时间、发育过程及成熟时间。结果表明:青虾幼虾发育到第10d(PL10)时促雄腺呈索状结构开始发育,随后经历增殖期(PL10)、合成期(PL13)和分泌期(PL19)3个发育阶段发育成熟,形成完整的促雄腺。青虾精巢和卵巢均在幼虾发育第13d(PL13)开始发育,精巢形成不规则排列的精原细胞,而卵巢生殖上皮开始分化为椭圆或多边形的卵原细胞。精巢经精原细胞期(PL13)、精母细胞期(PL16)、精细胞期(PL19)和精子期(PL22)4个发育阶段成熟,此时成熟的精巢生精小管内充满成熟的精子。卵巢经卵原细胞期(PL13)、初级卵母细胞期(PL16)、次级卵母细胞期(PL16)和卵子期(PL19)4个时期发育成熟,此时卵巢充满成熟卵子。从PL1到PL22,睾酮分泌量逐渐增加,维持在稳定水平至PL25后逐渐下降;从PL1到PL19,17β-雌二醇的分泌量逐渐增加,维持在稳定水平至PL25后逐渐下降。本研究显示:促雄腺与精巢发育过程均持续10d,但促雄腺早于精巢3d开始发育和成熟,这可为性别和生殖相关基因的筛选及性别调控机制的研究提供参考。     

泼洒糖蜜对池塘养殖罗氏沼虾生长和水质的影响

将单口面积1.33~2.33hm~2、环境相似的6口罗氏沼虾养殖池塘分为2组,3口池塘投喂蛋白质水平为35.4%的饲料,且每日适量泼洒糖蜜(试验组),3口池塘投喂蛋白质水平为41.9%的饲料,不泼洒糖蜜(对照组),常规管理。养殖期间,每隔15d取水样检测氨氮和亚硝态氮含量。156d的饲养结果表明,对照组和试验组罗氏沼虾的产量分别为6600kg/hm~2和6427.5kg/hm~2,降低饲料中的蛋白水平结合泼洒糖蜜不影响罗氏沼虾的生长(P0.05),养殖效益亦无显著差异(P0.05)。但试验组池塘水中氨氮较对照组低54.0%,亚硝态氮低21.0%,泼洒糖蜜显著降低了池塘氨氮和亚硝态氮含量(P0.05)。     

患铁虾综合征罗氏沼虾眼柄转录组研究

为探索罗氏沼虾(Macrobrachium rosenbergii)铁虾综合征(IPS)的分子机制,采用高通量测序平台(Illumina Hiseq-2500)分别对患IPS罗氏沼虾(IPS虾)和正常罗氏沼虾开展转录组测序,进行生物信息学分析。结果显示,高通量测序共获得56.42 G高质量数据,拼接后得到221 901条单基因序列(unigene),长度范围为201~30 985 bp,平均长度为1572 bp,N50长度为2867 bp,N90长度为646 bp。将单基因序列分别在Nr、Nt、Swissprot、KEGG、KOG、GO、PFAM数据库进行序列比对及功能注释,103 570条得到注释,其中,GO数据库注释到的单基因序列最多。差异表达分析显示,2003个基因在IPS虾眼柄中差异表达,包括1209个上调基因和794个下调基因,516个基因被注释到242条KEGG通路中,翻译、信号转导和免疫系统富集的差异基因数目最多。催乳素、雌激素、胰岛素、促性腺激素释放激素、胰高血糖素、催产素、谷氨酸能突触、血清素能突触等与生殖调控相关激素的代谢过程在IPS虾与正常虾眼柄之间存在差异。此外,一些已被证明在免疫反应中起重要作用的基因在IPS虾眼柄中显著上调,如血管内皮生长因子受体1、丝氨酸蛋白酶抑制剂6、C型凝集素、芳基硫酸酯酶B、酚氧化酶原激活酶2a、组织蛋白酶B、组织蛋白酶L、甲壳类抗菌肽4等。同时,注释到溶酶体、吞噬体、抗原处理与呈递、细胞凋亡、内吞作用等多条与免疫相关的途径,支持近期研究得出的罗氏沼虾IPS与病原感染相关的结论。本研究为解析罗氏沼虾IPS的成因和分子机制提供了数据支撑。     

复方中草药对青虾免疫功能的影响

将大黄、黄芩、党参、板蓝根等十余味中草药粉碎过筛后制成复方添加剂按1%、2%、3%的比例添加于基础饲料中制成颗粒药饵投喂青虾,测定青虾相关免疫指标的变化。结果表明,青虾血细胞吞噬百分比和吞噬指数、血清溶菌酶活力均显著提高。经嗜水气单胞菌攻毒后,各试验组的免疫保护率也明显提高。在各试验组中以2%的添加量的效果最好。试验证明,该添加剂可显著地提高青虾的免疫功能,能有效地预防嗜水气单胞菌的感染。     

Larval Development of the Giant River Prawn Macrobrachium rosenbergii at Different Ammonia Concentrations and pH Values

The effect of ammonia and pH levels on giant river prawn Macrobrachium rosenbergii larvae were evaluated to provide science-based information on safe levels of ammonia and pH for larviculture. Survival rate, developmental stage, and larval weight gain were determined for larvae kept in water with total ammonia (NH ₄-N)concentrations of 0, 1, 2, 4, and 8 mg\L and pH 7, 8, and 9. The trials were conducted in two phases: phase 1, larvae from stages I through VIII and phase 2, larvae from stage VIII until metamorphose. Oxygen consumption was determined for larvae in stages I and VIII at total ammonia concentrations of 0, 4, and 8 mg/L and pH 8. Survival rate up to stage VIII varied from 86 to 98% and did not differ for total ammonia concentrations in pH 7 and 8 and for 0 mg/L NH₄ N in pH 9. Survival rate was significantly lower (0–20%) for total ammonia concentrations from 1 to 8 mg/L (0.43–3.41 mg/L of unionized ammonia) in pH 9. Larval stage indexes (7.9–8.0 range) and weight gain (1.572–2.931 mg range) of larvae at the end of phase 1 of the experiment did not differ for the different ammonia concentration solutions, but were significantly lower in pH 9. In phase 2, no parameter differed among treatments for pH 7 and 8; however there was total mortality at pH 9 until 96h. Respiration rates diminished when larvae were exposed to total ammonia concentrations of 4 and 8 mg/L (0.28 and 0.55 mg/L of unionized ammonia), but development remained unaltered. Therefore, M. rosenbergii larvae tolerate high levels of total ammonia, while toxicity depends primarily on unionized ammonia concentrations. In addition, alkaline pH (9) acted directly on the larvae, curbing development and causing severe mortality. Larval tolerance to high ammonia and pH levels decreases for the last zoeal stages.     

鄱阳湖日本沼虾幼体发育及其同工酶研究

对鄱阳湖日本沼虾(Macrobrachiumnipponensis)幼体发育的5个阶段即Z0、Z1、Z3、Z5～Z6及Z9～Z11期进行观察,分析乳酸脱氢酶(LDH)、酯酶(EST)、苹果酸脱氢酶(MDH)同工酶谱带变化。结果表明,各期间幼体形态均有明显变化,幼体蜕皮次数随发育快慢不同,发育快的需9次蜕皮完成变态发育过程,慢的则需14次;LDH酶谱在5个发育阶段各有5条带,仅酶活性有强弱变化;EST酶谱不仅条带数从2条增加到9条,酶活性也有明显变化,EST1和EST3在5个阶段都出现,且酶活性逐渐升高;MDH酶谱共出现6条带,MDH1和MDH4分布延伸时间跨度大,酶活性高,可看作主带。因此日本沼虾变态发育过程中同工酶谱带随变态发育而变化,反映了不同组织器官细胞组成和不同组织中不同同工酶的活性和出现顺序,可作为鉴别种属特异性和发育阶段特异性的一项生化指标。     

罗氏沼虾气单胞菌MrM0602株的分离和致病性研究

从濒死的罗氏沼虾体内分离到菌株M rM0602,经形态学观察及API 20E鉴定系统鉴定为嗜水气单胞菌。人工感染健康罗氏沼虾,100%致死剂量为1.0×104cfu/m l,半致死剂量(LD50)为3.16×103cfu/m l。测定菌株M rM0602的胞外产物对小鼠和新西兰大白兔红细胞的溶血活性,表明胞外产物对小鼠红细胞有微弱的溶血活性,对新西兰兔红细胞无溶血活性。胞外蛋白酶活力测定显示,菌株M rM0602胞外产物具有很高的蛋白酶活力,且耐热性很强,65℃仍保持较高活性。常规药物敏感性试验表明,菌株M rM0602对环丙沙星和恩诺沙星高度敏感。     

Effects of Stocking Density on Nursery Production and Economics of the Freshwater Prawn,Macrobrachium rosenbergii

ABSTRACT

In temperate regions, post-larvae freshwater prawn, Macrobrachium rosenbergii, are grown to more advanced sizes in tanks prior to pond stocking. This intermediate stage of culture is referred to as the nursery period. Little research has been conducted on different management practices on juvenile prawn growth and survival during this 30-60 day period. Survival during the nursery stage has been highly variable and may be related to the cannibalistic behavior of juvenile freshwater prawn when cultured at high densities in the nursery. The objective of this study was to evaluate the effect of stocking density, relative to the provision of artificial substrate (number of prawns/m² of substrate), on growth, survival, and economic variables for freshwater prawn juveniles during nursery production. Post-larvae (0.01%0.00 g, n = 300) were stocked into nine 1900 L tanks, each provided with 20.5 m² of artificial substrate in the form of horizontal layers of black plastic mesh (10 mm) spaced 5 cm apart. Tanks were randomly assigned one of three prawn densities (215, 430, or 860 post-larvae/m² of substrate), which equated to 2.3,4.6 and 9.2 prawn/L, respectively. Juvenile prawn were fed a commercial trout diet (42% protein) at a percentage of body weight according to a feed rate table. Water quality was maintained using a flow rate of 8 L/min in each tank from a reservoir pond. Temperature was maintained at approximately 28°C using heat pumps. After 56 days there was no significant difference (P >0.05) in average weight of juvenile prawn stocked at the three densities (0 = 0.58%0.12 g, n = 9). Survival was significantly lower (P <0.05) for prawn stocked at 860 m² (62%) than in those stocked at 430/m² (78%) and 215/m² (94%), which were not statistically different (P >0.05). Even with reduced survival, the highest stocking density produced the greatest number of nursed juveniles based on both tank volume (5.5/l) and surface area (530/m²), at the lowest average cost.     

利用地热水进行罗氏沼虾越冬试验

利用地热水进行罗氏沼虾越冬试验。定时充气增氧,每天排污,日换水量15%～25%。约20d大换水一次,彻底排污。投喂人工配饵,消毒防病。实验证明:越冬水温21～25℃,溶氧保持3mg/L以上,能加速越冬对虾生长和发育,促进交尾,提高成活率。     

异型雄性罗氏沼虾遗传多样性的微卫星分析

利用微卫星DNA标记研究3种近亲交配形态型雄性罗氏沼虾遗传多样性,所用虾样均采自国家级(南宁)罗氏沼虾良种场,共150尾成年近亲交配的雄虾,其中蓝色长臂型、橙色长臂型、小个体型各50尾。研究结果表明,3组样品的生长发育差异显著,蓝色长臂型、橙色长臂型、小个体型各组中微卫星位点的总等位基因分别为24、28和22,3组样品所有位点的平均观察杂合度为0.474～0.556。其平均群体内近交系数为-0.0189,说明杂合子过剩,对比分析和群体间分化系数表明,3组雄虾遗传差异明显,最大遗传间距位于蓝色长臂型和小个体型,最小的遗传间距位于蓝色长臂型和橙色长臂型。研究结果将为罗氏沼虾优质父本选育和调控雄性遗传发育提供基础性遗传信息。     

氨态氮和亚硝态氮对日本沼虾酚氧化酶活力及血蓝蛋白含量的影响

在水温(24±1)℃下分别测定了日本沼虾暴露在0、2.2、4.2、8.7、17.8、36.6 mg/L氨氮和0、2.0、3.2、5.0、7.9、13.3 mg/L亚硝酸盐后血淋巴中酚氧化酶活力和血蓝蛋白含量的变化。试验结果表明,随着氨氮和亚硝酸盐浓度的升高,酚氧化酶活力先升后降,在氨氮质量浓度为8.7 mg/L、亚硝酸盐质量浓度为5.0 mg/L时,酚氧化酶活力最高。在氨氮质量浓度为2.2 mg/L、亚硝酸盐质量浓度为2.0 mg/L时,血蓝蛋白含量显著高于对照组。日本沼虾血淋巴中酚氧化酶活力和血蓝蛋白含量没有相关性。     

混养鲫鱼对罗氏沼虾种虾培育效果的影响

在3口面积各为0.3 hm^2、深度2.0 m的长方形池塘(1#、2#、3#)中放入体长0.7～0.8 cm的罗氏沼虾"南太湖2号"无特定病原种虾,密度为3.0×10~5尾/hm^2;养殖30 d时,2#、3#池塘分别混养质量3～4 g/尾的鲫鱼450、900尾,1#池塘不混养作对照组。经过近110 d的培育,1#、2#、3#池塘"南太湖2号"种虾的平均体质量分别达18.36、18.95、22.6 g,收获量分别达931.5、898.4、842.95 kg,饵料系数分别为1.36、1.53和1.71。试验结果表明,罗氏沼虾"南太湖2号"种虾培育池塘混养合理密度的鲫鱼,虽然影响种虾养殖产量,饲料系数增大,但培育出的种虾个体较大,饱满度较高,作为亲本可以为后期育苗工作的顺利开展打下良好基础。     

病毒、弧菌共同作用引发虾病的预防与治疗技术初探

通过用肉眼观察、镜检,并配合核酸探针法检测池塘对虾病虾的病毒、细菌感染情况,结果发现,由细菌引起的白斑病症较多,携带病毒和感染上病毒的虾数量很少。经分离培养和人工感染试验,证实1号菌是很强的致病菌。经药敏试验找出了首选药物阿莫西林。     

Apparent Control of Sexual Differentiation of Freshwater Prawn,Macrobrachium rosenbergii,Through Dietary Administration of Dopamine Hydrochloride

ABSTRACT

Dopamine at concentrations of 0.15, 1.5, and 15.0 mg/kg was included in two different formulated diets that were fed to recently metamorphosed postlarvae of the freshwater prawn, Macrobrachium rosenbergii, for 60 days. Dopamine, in the form of dopamine hydrochlo-ride, was incorporated into the formulated diets by either solubilizing it in alcohol, followed by submersion of diet and evaporation, or inclusion in a lipid premix ingredient. The alcohol evaporation method at 0.15, 1.5, and 15.0 mg/kg significantly increased the percentage of individuals without external male characteristics. This research is the first reported use of dietary administration of dopamine to crustaceans, and the first documentation of the apparent dietary control of gonadal development and sex differentiation in crustaceans. Replication of results, a histological examination of a large sample of individuals, and continued monitoring of those individuals fed the dietary dopamine are needed to verify the apparent effect.