Bacterial flora associated with the giant freshwater prawn Macrobrachium rosenbergii, in the hatchery system

The objective of this study was to understand the microbial flora associated with the hatchery system of giant fresh water prawn, Macrobrachium rosenbergii during an entire rearing cycle. Bacteriological and physico-chemical analysis was done for different samples of water, larvae, and Artemia. The total bacterial load in well water, seawater and inlet water varied from 10¹ to 10⁵ cfu ml^− 1 with higher counts seen in larval rearing tank (LRT) water. The Vibrio count ranged between 10¹ to 10³ cfu ml^− 1. Larval samples harboured a bacterial load of 10⁶ to 10⁷ cfu/10 larvae. The bacterial load in Artemia hatching water ranged from 4.90 × 10⁴ to 5.63 × 10⁶ cfu ml^− 1 while Artemia had a load ranging from 1.08 × 10⁷ to 2.09 × 10⁹ cfu g^− 1. Vibrio count in the LRT water ranged from 10¹-10³ cfu ml^− 1 while the count in larvae ranged from 10² to 10⁴ cfu/10 larvae. The bacterial genera were predominantly Gram-negative and comprised of Aeromonas spp., Pseudomonas spp., Vibrio spp. and Bacillus spp. and non-spore formers (NSF) were the dominant Gram-positive bacteria. This study documents the bacterial flora associated with Macrobrachium hatchery system during a regular normal run. Knowledge of the qualitative and quantitative aspects of bacterial flora in the hatchery would help to understand disturbances, if any, brought about during disease outbreaks.     

Effect of nitrite on larval development of giant river prawn Macrobrachium rosenbergii

The effects of ambient nitrite concentrations on larval development of giant river prawn Macrobrachium rosenbergii were evaluated. The trials were conducted in two phases: phase 1, larvae from stages I through VIII and phase 2, larvae from stage VIII until post-larvae. In both phases larvae were kept in water with nitrite (NO₂-N) concentrations of 0, 2, 4, 8 and 16 mg/L. Oxygen consumption was analyzed for larvae in stage II at nitrite concentrations of 0, 4, and 8 mg/L. Survival, weight gain, larval stage index and metamorphosis rate decreased linearly with increasing ambient nitrite concentration. However, there was no significant difference between larvae subjected to 0 and 2 mg/L NO₂-N. In phase 1, there was total mortality at 16 mg/L NO₂-N, while in phase 2 larval development stopped at stage X in this treatment. The oxygen consumption in stage II increased significantly at NO₂-N concentration from 0 to 4 mg/L, but there was no difference between 4 and 8 mg/L NO₂-N. In conclusion, increasing ambient nitrite up to 16 mg/L NO₂-N delays larval development, reduces larval growth rate and causes mortality, whereas no significant effect occurs for levels below 2 mg/L NO₂-N. However, the establishment of a general safe level of nitrite to M. rosenbergii hatchery may be difficult due to the great variability in larvae individual sensitivity.     

罗氏沼虾谷氨酰胺合成酶基因的克隆与表达

谷氨酰胺合成酶(glutamine synthetase, GS)是一种广泛分布在动植物体内的酶,并参与细胞多种代谢调控。在甲壳动物中,GS在能量代谢和渗透压调节过程中起着重要作用。实验克隆了罗氏沼虾GS基因。GS基因cDNA全长1 965 bp,开放读码框(ORF)为1 086 bp,编码361个氨基酸(aa),分子量大小为40.75 ku,等电点为5.81。进化树分析发现,罗氏沼虾GS基因与凡纳滨对虾、斑节对虾和中国明对虾GS聚为一支,氨基酸相似性达到95%。氨基酸多序列比对分析结果显示,罗氏沼虾GS属于无脊椎动物分支的GSⅡ群,有5个保守区域。实验对罗氏沼虾GS蛋白进行表达并制备了多克隆抗体。采用荧光定量PCR技术(qRT-PCR)和免疫印迹(Western blot)对罗氏沼虾蜕壳前后的不同组织GS表达进行检测。qRT-PCR结果显示,GS基因在所检测的8个组织中均有表达;蜕壳前,其相对表达量顺序为:肝胰脏肌肉胃肠鳃心脏脑血淋巴。蜕壳后和蜕壳前GS基因在组织中的差异表达比较结果显示,除了在肝胰脏表达下调外,其他7个组织都表达升高,其相对表达量的差异顺序为:脑鳃胃肠肌肉心脏血淋巴。Westernblot结果显示,蜕壳后GS蛋白在鳃和肌肉组织表达量上调,与其mRNA表达一致。此外,罗氏沼虾蜕壳后肝胰脏GS酶的活性和谷氨酰胺的含量下调,而其在鳃、肌肉和血淋巴中上调,结果与其基因表达一致。研究表明,GS基因在不同组织中表达的差异可能和罗氏沼虾的能量代谢、渗透压调节有关。     

Salinity effects on reproduction of giant freshwater prawn Macrobrachium rosenbergii (de Man)

Understanding the role of salinity in breeding and growth has the potential to enhance production of giant freshwater prawn, Macrobrachium rosenbergii. This study investigated the reproduction of females reared in salinities of 0, 6, 12, and 18 g L^− 1. Mean weight of females decreased with increased salinity (31.40 ± 1.54, 25.14 ± 1.16, 20.80 ± 0.81, and 16.62 ± 1.04 g at 0, 6, 12, and 18 g L^− 1, respectively). Larval production was delayed by 2 months in females reared in 12 g L^− 1 compared to 6 and 0 g L^− 1 and the cumulative number of berried females decreased with increased salinity. The number of larvae produced was positively correlated to weight of female. A larval production not only differed between salinity treatments, but also that larval production per gram of female differed between treatments. Females reared in lower salinity of 0 and 6 g L^− 1 produced larger numbers of larvae (12,155 ± 480 and 6519 ± 323, respectively) compared to 12 and 18 g L^− 1 (3751 ± 256 and 0, respectively). The number of larvae produced per gram of female was inversely related to the salinity levels (Y = − 37.54X + 685.65, n = 339, r² = 0.995, p < 0.05). Survival of larvae from females reared at 0 and 6 g L^− 1 was higher than those from females reared in 12 g L^− 1. This study clearly shows that female broodstock reared in lower salinity was larger, reproduced early, and produced more offspring than at higher salinity and this could significantly impact coastal prawn culture where seasonal fluctuation of salinity in the hatchery is common.     

罗氏沼虾几丁质酶3B基因的克隆及其在蜕皮周期中的表达

罗氏沼虾的生长发育及繁殖都与蜕皮紧密相关。几丁质酶是甲壳动物在蜕皮过程中最重要的酶之一。本研究对罗氏沼虾蜕皮周期外观特征和腹肢刚毛进行了观察描述,克隆并分析了罗氏沼虾的几丁质酶Ⅲ基因(MrChi3B),制备了几丁质酶Ⅲ蛋白的多克隆抗体,研究了其在蜕皮周期中的表达。罗氏沼虾几丁质酶基因cDNA的开放阅读框为1 143 bp,编码380个氨基酸,大小为41.91 ku。通过氨基酸序列比对发现,MrChi3B与日本沼虾几丁质酶基因(Chi3B)的同源性最高,为94%。MrChi3B含有一个糖苷键水解酶家族18的催化域。实时定量PCR (qRT-PCR)和蛋白印迹法(WB)检测结果显示,MrChi3B在罗氏沼虾多个组织中均有表达,但是不同组织中的表达有差异。在蜕皮期之后,其在胃、表皮和肌肉中的表达量显著上升。在胃中其表达量在蜕皮间期达到最高;在表皮和肌肉中其表达量在蜕皮后期达到最高;肠中的表达量在蜕皮前期和蜕皮期有所上升;眼柄期的表达量在所有蜕皮期都偏低。本研究结果为进一步研究几丁质酶的功能提供参考依据。     

饲料中n-3 PUFA/n-6 PUFA比值对罗氏沼虾幼虾生长性能和抗氧化能力的影响

为探讨饲料多不饱和脂肪酸n-3 PUFA/n-6 PUFA比值对罗氏沼虾幼虾生长性能、虾体肌肉组成、抗氧化能力、血清生理指标以及消化能力的影响,实验设计了n-3 PUFA/n-6 PUFA比值分别为0.37 (D1)、0.59 (D2)、0.93 (D3)、1.51 (D4)和4.38 (D5)的5种等氮等脂饲料饲喂罗氏沼虾幼虾8周,每组设4重复,每个重复40尾虾 。结果显示,饲料n-3 PUFA/n-6 PUFA比值对罗氏沼虾幼虾存活率 (SR)无显著影响;实验虾终末体重 (FW)、增重率 (WGR)和特定生长率 (SGR)随饲料n-3 PUFA/n-6 PUFA比值增加先升后降,均在D3组最高;且D3组虾有最大的肝胰腺蛋白酶活性及脂肪酸合成酶活性。虾体肌肉粗蛋白质、水分和灰分含量不受饲料n-3 PUFA/n-6 PUFA比值影响,但总脂肪含量在D3组显著高于其他组;各组虾体肌肉的n-3 PUFA/n-6 PUFA比值的变化趋势与饲料的n-3 PUFA/n-6 PUFA比值变化趋势呈正相关。随饲料n-3 PUFA/n-6 PUFA比值增加,实验虾血清和肝胰腺中超氧化物歧化酶 (SOD)活性、总抗氧化能力 (T-AOC)和血清铜蓝蛋白 (CP)含量均呈现先升后降趋势,并在n-3 PUFA/n-6 PUFA比值为0.93~1.51时达到最大,但丙二醛 (MDA)含量持续上升;D1组血清总胆固醇 (T-CHO)和甘油三酯 (TG)含量显著高于其他组;血清谷草转氨酶 (AST)和谷丙转氨酶 (ALT)活性先降后升,且D3组最低。研究表明,饲料适宜的n-3 PUFA/n-6 PUFA可显著提升罗氏沼虾生长性能和抗氧化能力,对增重率和特定生长率进行折线回归,建议罗氏沼虾幼虾饲料中最适n-3 PUFA/n-6 PUFA比值为0.86~0.94。     

Effects of dissolved oxygen on hemolymph parameters of freshwater giant prawn, Macrobrachium rosenbergii (de Man)

Profiles of changes in physiological parameters of freshwater giant prawns, Macrobrachium rosenbergii, exposed to various dissolved oxygen (DO) levels of 7.75, 4.75, 2.75, and 1.75 mg l⁻¹ are reported. The parameters involved in osmoregulation and oxygen transport were monitored for a 6-day period. Notable depressions in hemolymph osmolality, Na⁺, K⁺, and Cl⁻ contents were observed within 24 h after exposure to hypoxia at 2.75 and 1.75 mg O₂ l⁻¹, and thereafter remained at rather steady levels, which were significantly lower than those under normoxic conditions (4.75–7.75 mg O₂ l⁻¹). The extent of depression of osmotic-related constituents, hemolymph osmolality and Cl⁻ in particular, increased with decreased DO.

Oxyhemocyanin constituted 65.46–65.84% of total hemolymph proteins under the various DO levels examined; both hemolymph oxyhemocyanin and proteins showed notable elevations 24 h after exposure to hypoxic conditions, and reached the highest and constant level by 48 h after exposure. The compensatory responses of prawns to reduced O₂ were manifested by increased O₂ uptake through augmentation of hemocyanin, which results in enhancement of oxygen binding capacity of the hemolymph. In the same period, a significant surge of the respiratory products, PCO₂ and HCO₃⁻, was also demonstrated 6 h after exposure to hypoxic conditions which resulted in hemolymph alkalosis. These processes likely resulted in an increase in water influx and consequent declines in hemolymph osmolality and ion composition. Furthermore, hyperventilation and respiratory alkalosis, indicated by increased oxyhemocyanin and pH, respectively, were found to be predominant responses of M. rosenbergii to hypoxic stress.     

A comparative study on larviculture techniques for the giant freshwater prawn, Macrobrachium rosenbergii (de Man)

Comparative studies of larviculture of Macrobrachium rosenbergii (de Man) were made for three systems: (1) the static water system with partial changes of water, (2) the closed recirculating water system with a separate subsand filter unit, and (3) the closed recirculating water system with a subsand filter inside the rearing tank. The mean production of juveniles per unit volume of water of these three systems was comparable (3.2, 1.9, and 2.9, respectively). Production fluctuated more in the static system than in the closed recirculating system. A significant asymptotic relationship between the stocking density and the survival of prawn larvae was observed in the first type of the closed recirculating system; higher stocking density resulted in lower survival. A stocking density of 20 larvae/l in the closed recirculating system was considered to be good for growth and survival. The intermittent high nitrite level was a problem encountered in the static system but not in the closed recirculating system. The operational costs of these three systems were discussed.     

罗氏沼虾养殖群体表型性状间的相关性及类群差异分析

为研究罗氏沼虾(Macrobrachium rosenbergii)养殖群体表型性状间的相互关系及不同类群间的性状差异,本研究对人工养殖的罗氏沼虾雌、雄及“老头虾”3个类群共490个个体的体重、体长、全长等12个表型性状进行了相关分析、形态性状对体重影响的通径分析等。结果表明, 3个类群除上、下额刺数2个可数性状外,其他9个可量形态性状与体重都呈极显著的正相关(P<0.01);影响体重的关键形态性状在3个类群中各不相同,但体长、头胸甲长、第二步足长在3个类群中均是影响体重的关键指标;分别建立了各类群体重与关键形态性状的多元回归方程。聚类分析和主成分分析结果表明,雌、雄2个类群表型上更为相似,“老头虾”与雌性个体的差异最大。腹部轮廓特征(包括腹长、腹宽、腹高)、全长、体长及第二步足长对3个类群的差异贡献较大。雌性个体腹部发达,“老头虾”则第二步足尤为发达。本研究结果可为罗氏沼虾的选择育种、基础生物学及行业标准的制定提供数据资料。     

Observations on the breeding and growth of the giant freshwater prawn Macrobrachium rosenbergii (de Man) in the laboratory

Three male and 20 female prawns, Macrobrachium rosenbergii (de Man), were observed for 390 days. They were maintained in brackish water (5%. salinity) at 28°C which was recycled continuously through a percolating biological filter. Illumination was artificial and did not exceed 10 lm/ft² (approx. 0.1 m²) at the water surface. Mating readily occurred in the experimental tanks (48 × 28 × 25 cm deep). Eggs were incubated for 20 days; the mean number of larvae per brood was 24 000 (range 50–98 100). Over 750 000 larvae were hatched during the experiment. Larger females had proportionately larger broods and larvae from seven broods were cultured to the post-larval stage at intervals throughout the experimental period, which demonstrated their viability.The increase in length of the adults at each moult was constant (arithmetic growth) and did not alter when ova were maturing in the ovary.Prawns achieved larger mean length increments after the environmental conditions were improved. The moulting frequency was very variable and, with one exception, did not change proportionately with length or age of the prawns.Females grew from 115 to 205 mm and males from 145 to 230 mm total length.Three females spawned more than four times in successive intermoult periods, and one produced viable larvae five times in succession.Two of the males sired viable larvae four and seven times respectively during one intermoult period.     

The isosmotic concept in relation to the aquaculture of the giant prawn, Macrobrachium rosenbergii

Several workers have suggested that maximal growth of an organism should occur in isosmotic media, since the animals would not be expending energy in doing osmotic work. Growth and osmoregulatory studies, however, show that this is not true in the giant prawn, Macrobrachium rosenbergii. This paper attempts to explain the phenomenon.     

Effect of probiotics on immune competence of giant freshwater prawn Macrobrachium rosenbergii

This study evaluates the effect of probiotics, Zymetin, on the immune efficiency of Macrobrachium rosenbergii juvenile against pathogenic Vibriospp. and Aeromonasspp. The experiment was conducted in glass aquaria with same level of feeding under different treatments, that is, negative control (Cn), positive control with Vibrio spp. (Cv) and Aeromonasspp. (Ca), prawn juveniles fed with probiotics (Zymetin) at 5 g/kg of feed but without pathogen (T1), probiotic fed prawn challenged with Vibrio spp. (T2) and Aeromonasspp. (T3). The results demonstrated that T1 showed higher survival rate, total haemocyte count, non‐granular haemocyte count, phagocytic activity, and clearance efficiency compared to other treatment groups. In contrast, decreased number of small and large granular haemocyte was observed in T1. Despite that, THC was found to be significantly different (p < 0.05) among all the controls and lowest was in Cv, followed by Ca. Besides, phagocytic activity and clearance efficiency against Vibrio spp. and Aeromonasspp. decreased significantly in Cn, Ca, and Cv fed M. rosenbergiijuvenile, while the values were found to be higher in T1 followed by T2 and T3, in that order. In addition, the challenge test showed an increasing trend of total and beneficial bacterial density as well as declining trend of some harmful bacteria in the water and gut of prawn in all the groups tested (p < 0.05).     

Evaluation of vitamin C-enriched Artemia nauplii for larvae of the giant freshwater prawn

The effect of high levels of ascorbic acid (AA) delivered through enriched live food has been verified through the successful culture of larval giant freshwater prawn, Macrobrachium rosenbergii. Two successive feeding trials were set up using a control (550 g AA g^–1 DW) and two different AA-enrichment levels in Artemia (1300 and 2750 g AA g^–1 DW). Under standard culture conditions, no differences in growth nor survival could be observed demonstrating that the nutritional requirements are below 550 g AA g^–1 DW, which is the normal level occurring in freshly-hatched Artemia. However, a significantly positive effect could be demonstrated on the physiological condition of the postlarvae, measured by means of a salinity stress test, when vitamin C-boosted live food was administered. Since the AA levels in the predator larvae are linked with the enrichment levels in the live prey, it may be assumed that a positive influence on stress resistance was caused by feeding vitamin C-enriched Artemia. It is expected that under suboptimal conditions, supplementation of high vitamin C levels might also enhance production characteristics.     

Preparation of monoclonal antibody against Macrobrachium rosenbergii Nodavirus and application of TAS-ELISA for virus diagnosis in post-larvae hatcheries in east China during 2000-2004

“Whitish muscle disease” of Macrobrachium rosenbergii, also called “whitish disease” or “white tail disease”, is a new serious epizootic disease that has occurred in recent years in giant freshwater prawn culture regions, mainly in southern China. This disease occurred in post-larvae 3-5 days to 3 weeks after desalting. Clinical signs include the development of white spot in muscles or milky muscles throughout the body, causing serious loss in few days, with a mortality rate of 40-90%. A 26-27 nm icosahedral non-enveloped virus, identified as M. rosenbergii Nodavirus (MrNV), was confirmed as the aetiological agent. Twelve hybridomas strongly secreting monoclonal antibodies (Mabs) against MrNV were shown to be specific for MrNV and reacted with MrNV 42 kDa coat protein by Western blot. A triple antibody enzyme-linked immunosorbent assay (TAS-ELISA) was developed and shown to be a useful diagnostic tool for MrNV infection.     

罗氏沼虾胞质锰超氧化物歧化酶的功能

为了解超氧化物歧化酶 (SOD)分子生物学特性和免疫功能,实验克隆了罗氏沼虾胞质锰SOD基因 (MrcMnSOD),制备多克隆抗体,并分析在嗜水气单胞菌感染下该基因的表达模式。结果显示,嗜水气单胞菌感染可显著诱导MrcMnSOD在转录水平和蛋白质水平进行高表达。为探究MrcMnSOD参与免疫应答的机制,进一步的抑菌实验表明,该蛋白质可显著抑制3种革兰氏阴性细菌 (大肠杆菌、副溶血性弧菌、嗜水气单胞菌)和2种革兰氏阳性细菌 (无乳链球菌、金黄色葡萄球菌)的生长,且抑制作用与蛋白质浓度的关系不显著。研究表明,MrcMnSOD可能作为一种免疫相关分子参与免疫应答反应。本研究初步探讨了MrcMnSOD的免疫生物学功能,旨在为深入研究罗氏沼虾SOD的功能奠定相关基础。     

Classification of differentiating oocytes during ovarian cycle in the giant freshwater prawn, Macrobrachium rosenbergii de man

Based on the light microscopic observations of cells' sizes, chromatin patterns, amount of lipid droplets and yolk granules, the female germ cells could be classified into four different phases, which include 1) oogonia (Oog), 2) primary oocytes (pOc), 3) secondary oocytes (sOc), and 4) mature oocyte (mOc). Oog are small oval-shaped cells with irregular-shaped nuclei sizing 4–6 μm in diameter. They rest on the connective tissue germinal cord at the tip of each ovarian pouch (lobule). Oogonia increase their number through mitotic division, and the daughter cells move into ovarian pouch where they undergo first meiotic division to become primary oocytes, which have various steps of 1st meiotic prophase accumulating at the innermost zone of the ovarian pouch. The primary oocytes are small oval-shaped cells (8.5–10 μm in diameter) with large nuclei containing chromatin in various states of condensation that finally transform into chromatids. Their nuclei are surrounded by thin rim of faint blue-stained cytoplasm. The secondary oocytes derived from 2nd meiosis and comprise five steps: Oc1 and Oc2, classified as previtellogenic oocytes, Oc3 and Oc4, classified as vitellogenic oocytes, and mature oocyte (mOc) The zones of ovarian pouch are defined based on the accumulation of various steps of developing oocytes, namely, oogenic, previtellogenic, vitellogenic and mature zones, respectively. The ovarian cycle is divided into five stages based on the number and types of oocytes present in each stage. Stage 0 and I are spawn and spent stages. Stage II and III are proliferative and premature stages, while stage IV is mature stage. During ovarian stage I, each ovarian pouch contains primarily oogonia, primary oocytes, Oc1 and a few Oc2. In stage II, the pouch contains mainly Oc2 and Oc3, while in stage III the predominant cells are Oc4. Mature oocytes appear synchronously, in stage IV. The ovulating mature oocytes pass through the thin disrupted wall of ovarian pouch into subcapsular space, that leads into the oviduct situated on the ventro-lateral side of the ovarian lobe. At spawning, the ovarian pouches break down and only connective sheaths and hemolymph sinuses remain. The germinal cords and islets of oogonia remain in the central area of stage 0 ovary. The ovarian capsule, including the muscular layer, becomes attenuated as the ovary progresses from stage 0 to IV. The hemolymph vessels become highly convoluted in the central area of the ovary, and they branch radially into smaller hemolymph sinuses around each oogenic pouch.     

Prey ingestion and live food selectivity of marble goby (Oxyeleotris marmorata) using rice field prawn (Macrobrachium lanchesteri) as prey

Marble goby (Oxyeleotris marmorata), a carnivorous fish native to freshwater in Asia-Pacific region, is a high-valued species in many Asian countries. The present study consisting of three experiments was conducted to determine the appropriate density, size and ingestion time of marble goby fingerlings on rice field prawn (Macrobrachium lanchesteri) as prey. Results showed that the ingestion rate of marble goby fingerlings (0.26-1.43 g size) on prawns (9-12 mm in length) had an asymptotic pattern peaked at a predator to prey ratio of 1:20. Marble goby fingerlings did catch prawns during both daytime (07:00-17:00 h) and nighttime (17:00-07:00 h), but the ingestion peaked during nighttime period. The hourly ingestion rate during nighttime was significantly higher than that during daytime for the smaller-size marble goby fingerlings of 0.26-0.87 g, while there was no significant difference in the hourly ingestion between nighttime and daytime for the larger-size marble goby fingerlings of 1.43 g. The daily ingestion of marble goby fingerlings on prawns (mg consumed prawns/goby/day) increased significantly with increasing average weights (g) of marble goby fingerlings (Y = 27.6 + 47.8X, n = 30, r² = 0.86, p < 0.05). Results also showed that the preferred prawn size of marble goby fingerlings of average weights of 0.26-4.18 g was small (9-12 mm) with positive electivity index of 0.19-0.33, and the shift from small- to medium-size (12-14 mm) prawns was slow. The present study has demonstrated the feasibility of using rice field prawns as live foods to nurse marble goby fingerlings.     

壬基酚(NP)对罗氏沼虾幼虾生长和性别分化的影响

采用外部形态观察和性腺组织学连续切片方法,确定罗氏沼虾幼虾的性别分化时期,并探讨壬基酚(NP)对罗氏沼虾幼虾(体长约1.5 cm)生长和性别分化的影响。结果表明,罗氏沼虾仔虾后15 d出现生殖原基;仔虾后24 d,部分仔虾第四至第五步足基部明显凹陷,腹甲出现皱折和突起,并出现生殖管;仔虾后45 d,部分仔虾第二游泳足内侧出现雄性附肢,性腺明显区分为精巢和卵巢。处理时间短于15 d,NP对罗氏沼虾幼虾生长无显著影响;但处理超过20 d,NP以明显的剂量依存关系显著抑制罗氏沼虾体长和体重(P<0.05)。实验结束时(30 d),对照组个体均已完成性别分化,精巢和卵巢内分别出现大量精原细胞和卵原细胞,但NP处理组无法鉴定性别的幼虾比例随NP剂量增加而增加。研究表明,罗氏沼虾幼虾外部性征出现略早于内部性征的分化,早期性别分化发生在仔虾变态后21～45 d;此外,NP可能通过干扰蜕皮及卵黄蛋白原合成从而抑制罗氏沼虾幼虾生长和精巢发育。     

大豆抗原蛋白对罗氏沼虾生理生化及免疫的影响

为探讨大豆抗原蛋白与豆粕饲用效果的关系,本研究配制4种等氮等能的实验饲料,以含有21.5%鱼粉、15.45%豆粕和15%发酵豆粕的实用饲料为基础饲料(T15组),用发酵豆粕完全替代T15组中的豆粕(TF组),比较发酵豆粕对豆粕的替代效果;再以鱼粉、酪蛋白和大豆分离蛋白为主要蛋白源分别配制与T15、TF组相近抗原蛋白含量的半纯化饲料AP5、AP0(抗原蛋白含量分别为5%和0%)组,探究抗原蛋白的生物学效应。选用初始体重为(0.17±0.02) g的罗氏沼虾(Macrobrachium rosenbergii)在室内水泥池网箱中进行为期64 d的养殖实验。结果显示:两种系列饲料中,大豆抗原蛋白含量的降低对罗氏沼虾生长、血清生化指标及免疫相关基因表达有不同的影响。在实用饲料组中,随着大豆抗原蛋白的降低,罗氏沼虾的增重率、肝胰腺蛋白酶活力、血清白蛋白、总蛋白、鳃Toll受体mRNA、NF-κB mRNA和肝胰腺HSP70 mRNA相对表达量都显著降低(P0.05),而血清超氧化物歧化酶活力显著升高(P0.05);在纯化饲料组中,随着大豆抗原蛋白的降低,罗氏沼虾的增重率、血清超氧化物歧化酶活力和丙二醛含量、肝胰腺HSP70mRNA相对表达量没有显著性差异(P0.05),肝胰腺淀粉酶活力、蛋白酶活力、血清白蛋白、总蛋白都显著降低(P0.05),而肌肉粗灰分、血清谷草转氨酶活力、鳃Toll受体mRNA、NF-κBmRNA相对表达量显著升高(P0.05)。比较实用饲料组与半纯化饲料组的结果可知,罗氏沼虾对实用饲料中5%的大豆抗原蛋白有一定的耐受性,并且有利于其生长和健康,推测大豆抗原蛋白与豆粕中的其他抗营养因子间存在有益的联合效应,显著降低其中的大豆抗原蛋白含量则不利于蛋白合成代谢和肝胰腺健康,导致生长性能下降。建议在实际生产中发酵豆粕和豆粕按一定比例混合使用。     

罗氏沼虾不同养殖模式对水体浮游生物的影响

通过围隔实验比较罗氏沼虾不同养殖模式对水体浮游生物的影响。实验设置6种养殖模式:罗氏沼虾单养(MP)、罗氏沼虾+浮萍(水面覆盖率5%)(PP)、罗氏沼虾+鲢(PF)、罗氏沼虾+背角无齿蚌+鲢(PMF)、罗氏沼虾+背角无齿蚌+浮萍(PMP)、罗氏沼虾+背角无齿蚌+浮萍+鲢(PMPF)。养殖64 d后,测定不同模式中浮游植物和三大类浮游动物(轮虫、枝角类及桡足类)的种类和数量。结果显示,上述6种模式中浮游植物共同优势种有4种,但优势度指数最大的浮游植物不同,MP组是锥囊藻属,有浮萍的PP和PMP组均为细小平裂藻,混养鲢的PF、PMF和PMPF组均为针杆藻。不同养殖模式无共同的浮游动物优势种。养殖模式对浮游生物密度具有显著影响,PF组浮游植物密度最高,MP组浮游植物密度最低,PF组浮游植物密度比MP、PP和PMP组分别高78%、53%和61%。相反,浮游动物密度MP组最高,PF组最低。混养鲢的PF、PMF和PMPF组浮游动物密度显著低于其他3组。研究表明,罗氏沼虾养殖中混养鲢可增加浮游植物密度而降低浮游动物密度,浮萍和鲢影响池塘优势种。