基于珊瑚骨基质的凡纳滨对虾内循环养殖系统的构建与运行效果

采用珊瑚骨作为生物膜载体,利用海水素配制人工海水,构建盐度为15‰(海水)和5‰(淡水)的两个凡纳滨对虾内循环养殖系统,通过添加硝化细菌菌剂和氮源,分别用8 d和13 d建立硝化功能。按照500尾/m~3密度投入虾苗后,海水系统和淡水系统分别运行97 d和83 d。在运行期间淡水和海水系统养殖水体氨氮浓度始终维持在较低水平,平均浓度分别为(0.015±0.008) mg/L和(0.014±0.008) mg/L;在海水系统运行前60 d,亚硝氮浓度维持在较低水平,在60～90 d,亚硝氮浓度呈缓慢上升趋势,在90 d后,海水系统亚硝氮浓度开始快速增加,最终达到3.43 mg/L;淡水系统在运行前40 d亚硝氮浓度维持在较低水平,40 d后开始小幅上升,运行至70 d后,亚硝氮浓度开始快速增加,最终达到0.52 mg/L。最终海水系统和淡水系统凡纳滨对虾存活率分别为51.5%和48.5%。     

人工培养条件下环境因子对微小小环藻生长的影响

在实验室培养条件下,研究了温度、光照、盐度、pH值和氮、磷、铁、硅等营养盐对微小小环藻(Cyclotella caspia)生长繁殖的影响。实验结果表明:微小小环藻生长的适宜温度为10～40℃,最适温度为30～35℃;适宜光照为500～10 000 lx,最适光照为2 000～8 000 lx;适宜盐度为2～85,最适盐度为25～50;适宜pH值为4～9,最适pH为8.0～8.5。筛选出微小小环藻培养液配方是每立方米海水加入(NH2)2CO-N 30g,NaH2PO4-P 1 g,FeC6H5O7-Fe 0.1 g,Na2SiO3-Si 25 g。     

凡纳滨对虾养殖池塘硝化细菌的分离鉴定及脱氮效果研究

利用平板稀释涂布法和平板划线分离纯化法,自凡纳滨对虾养殖池塘中分离得到5株细菌,经过形态观察、生理生化特性以及16S rDNA序列分析,对分离菌株进行鉴定,同时对其硝化功能和氨氮去除特性进行研究。试验结果显示,分离得到的5株细菌分别为门多萨假单胞菌、嗜胺甲基杆菌、放射根瘤菌、施氏假单胞菌和氧化微杆菌。对其净化效果进行验证可知,门多萨假单胞菌和施氏假单胞菌对氨氮的去除率超过85%,分别为88.83%和91.73%;对亚硝态氮的去除率超过90%,分别为95.08%和97.39%;对硝态氮的去除率超过90%,分别为90.49%和91.22%。以上两种菌株有较高的氨氮和亚硝态氮去除效果,具有潜在的应用价值。     

硝态氮对凡纳滨对虾血清免疫相关酶活性的影响

以体质量(8.68±1.62)g、体长(9.31±0.65)cm的凡纳滨对虾(Litopenaeus vannamei)为研究对象,通过48h胁迫,研究了不同质量浓度(4.24 mg·L-1、35 mg·L-1、70 mg·L-1、105 mg·L-1和140 mg·L-1,以氮质量浓度计)硝态氮(NO-3-N)对对虾血清中酚氧化酶(PO)、超氧化物歧化酶(SOD)、碱性磷酸酶(AKP)、酸性磷酸酶(ACP)、溶菌酶(LSZ)、一氧化氮合酶(NOS)等活性的影响,探讨凡纳滨对虾养殖中NO-3-N的安全质量浓度。结果表明,质量浓度不高于140 mg·L-1的NO-3-N对凡纳滨对虾血清中SOD、PO、AKP、ACP、i NOS、TNOS、LSZ等酶活性的作用主要表现为短期影响;随实验时间增加,组间差异减小;第48小时除35 mg·L-1组外,其他组的诱导型一氧化氮合成酶(i NOS)活性差异并不显著。     

天津市周边地区养殖凡纳滨对虾肠道菌株的分离鉴定及耐药性分析

随着抗生素在水产养殖业中的普遍应用,细菌耐药性成为环境和食品安全的主要问题。本文旨在通过对天津地区养殖凡纳滨对虾体内细菌的耐药信息进行分析统计,为凡纳滨对虾养殖业的疾病控制和提高水产品安全性提供理论指导。本研究对天津市津南区、汉沽区及西青区等对虾养殖基地取样的凡纳滨对虾进行肠道微生物的分离培养,抗生素药敏实验以及抗生素抗性基因的检测。共获得菌株1 219株,选取其中有代表性的215株菌株进行鉴定,结果显示,所鉴定菌株属于56种不同种属,主要以芽孢杆菌、副溶血性弧菌、葡萄球菌、肺炎克雷伯菌、席罗弧菌、溶藻弧菌、格氏乳球菌、希瓦氏菌、霍氏肠杆菌、库特氏菌、施氏假单胞菌、荚膜红细菌、液化沙雷氏菌、不动杆菌、假单胞菌、假交替单胞菌及霍乱弧菌等为主。所鉴定菌株的抗生素药敏实验分析结果发现,凡纳滨对虾肠道微生物普遍存在抗生素抗性,其中β-内酰胺类的氨苄西林、糖肽类的杆菌肽、喹诺酮类的罗美沙星和磺胺类的磺胺抗药性最为明显。因此本文进一步对其中耐药种类较多且耐药性较强的82株细菌对四大类抗生素(链霉素类、磺胺类、四环素类和青霉素类)的抗性基因(str A-str B,sul II,sul II,sul III,tet A,tet C,tet G,blao XA-1,bla PSE-1,bla TEM)以及整合子(in T1和in T2)携带情况进行检测。研究结果表明,所检测耐药菌株均携带抗性基因,其中磺胺类抗性基因的检出率最高,可达63%,其次是青霉素类抗性基因,检出率为61.4%,整合子基因平均检出率为42.1%,四环素类相对较少为35.4%,且四环素类只检测出tet A抗性基因,其中67.1%的菌株存在多种耐药基因。     

颤藻对凡纳滨对虾生长和免疫相关酶活力的影响

颤藻(Oscillatoria sp.)是对虾养殖水体中的一种常见蓝藻。通过在凡纳滨对虾养殖水体中投加不同浓度的颤藻,测定凡纳滨对虾的成活率与体长、体重,以及不同颤藻浓度下凡纳滨对虾的免疫相关酶活力,研究颤藻对凡纳滨对虾生长和免疫酶活力的影响。结果表明:颤藻对对虾的成活率、体长和体重有显著影响(P<0.05),随藻浓度增加对虾的成活率、体长和体重呈下降的趋势。颤藻对对虾超氧化物歧化酶(SOD)、过氧化物酶(POD)、酚氧化酶(PO)、碱性磷酸酶(ALP)、谷胱甘肽S-转移酶(GST)、抗菌活力影响显著(P<0.05),低浓度的颤藻诱导酶活力增大,当浓度大于12.5 mg/L时,其酶活力呈下降的趋势。当颤藻浓度为2.5 m g/L和12.5 mg/L时,GST、ALP活力分别达到最大值(39.05 U/mg、73.62 U/g)。以上研究结果说明,颤藻具有一定的毒性,低浓度的颤藻可诱导酶活性增加,高浓度的颤藻能抑制酶的活性和对虾生长。为了保证凡纳滨对虾的健康养殖,养殖水体中的颤藻浓度应该控制在12.5 mg/L以下。     

颤藻浓度和水温对凡纳滨对虾响应颤藻粗提液毒性的影响

文章研究了绿色颤藻(Oscillatoria chlorina)不同成分对凡纳滨对虾(Litopenaeus vannamei)的毒性,分析了颤藻浓度与水温对对虾响应粗提液毒性的影响,并初步检测了颤藻粗提液所含的毒素。结果显示,湿藻质量浓度为5.04 mg·m L~(-1)的颤藻细胞及其粗提液可导致93.72%以上的对虾死亡,藻细胞培养液对对虾无致死效应。用湿藻质量浓度高于2.52 mg·m L~(-1)的粗提液注射对虾,对虾死亡率超过86.67%,随质量浓度低至0.504 mg·m L~(-1),对虾死亡率降至2.22%,差异显著(P0.05);水温显著影响了粗提液对对虾的致死效应(P0.05),湿藻质量浓度为2.52 mg·m L~(-1)的粗提液在22℃水温条件下可引起20.0%对虾死亡,当温度升至34℃,对虾死亡率增加至97.8%;以微囊藻毒素(Microcystin-RR/YR/LR,MC-RR/YR/LR)作标准品,使用高效液相色谱法检测颤藻粗提液毒素,结果在粗提液中均未检出RR、YR、LR等3种常见的微囊藻毒素。结果表明,绿色颤藻可致对虾急性死亡,其主要有害成分是颤藻粗提液所含的非MC-RR/YR/LR等藻毒素,对虾对颤藻粗提液毒性的响应与颤藻浓度和水温显著相关(P0.05)。     

生物絮团对凡纳滨对虾养殖过程中氨氮和亚硝酸氮含量的影响

本实验以非生物絮团养殖模式作为对照,研究了生物絮团凡纳滨对虾养殖模式中,水质因子氨氮和亚硝酸氮的变化规律。结果表明:试验组的生物絮团沉积量至第35天达到峰值(15.93±0.31)m L/L,而后保持相对稳定状态,对照组的生物絮团量一直处于极低水平(1.5 m L/L),两组之间差异显著(P0.05);对照组氨氮含量至第35天达到峰值(1.05±0.19)mg/L,试验组氨氮含量增加缓慢,至第60天时仅为(0.37±0.04)mg/L,显著低于对照组(P0.05);在实验的前15天,实验组和对照组的亚硝酸氮含量无显著差异(P0.05),随后试验组亚硝酸氮含量增速减慢并趋于稳定,而对照组则直线上升,对照组亚硝酸氮含量显著高于试验组(P0.05)。     

凡纳滨对虾中发光坎氏弧菌的分离、鉴定及致病性

为研究凡纳滨对虾死亡的致病机理,实验采用TCBS培养基从濒死凡纳滨对虾肝胰腺分离到浅绿色、直径3～7 mm的发荧光菌落;革兰氏染色为阴性短杆菌;经API 20E鉴定,分离株PvL-1与坎氏弧菌参考菌株CAIM249相似度为90％;fitZ序列与坎氏弧菌CP020076相似度为99.31％;gapA序列与坎氏弧菌EF59...     

亚硝态氮慢性胁迫对凡纳滨对虾体成分和糖代谢的影响

为研究亚硝态氮(NO2–-N)慢性胁迫对凡纳滨对虾(Litopenaeus vannamei)[(2.03±0.33) g]生长、摄食、体成分和糖代谢的影响,实验设置0 (对照组)、8 (N8组)、15 (N15组)和30 mg/L (N30组) NO2–-N浓度组,进行36 d的NO2–-N慢性胁迫实验。结果显示,随着NO2–-N浓度升高,对虾的终末体重、特定生长率、增重率和日摄食率均呈显著下降趋势。N8组和N15组对虾的血糖和肝胰腺中糖原含量在实验中期高于对照组,但最终均低于对照组,而肌肉中糖原含量在实验期间始终低于对照组。胁迫组凡纳滨对虾肌肉中的己糖激酶活力低于对照组;而肝胰腺己糖激酶活力和丙酮酸激酶活力均高于对照组,同时,肝胰腺乳酸含量与乳酸脱氢酶活力呈先上升后下降的趋势,最终与对照组相比无显著差异。终末体成分中,N30组的粗脂肪含量显著低于对照组。结果表明,NO2–-N慢性胁迫会降低对虾食欲,减缓凡纳滨对虾的生长,机体糖代谢调节可作为对虾应对NO2–-N慢性胁迫的短期响应,对虾对脂质的利用在应对NO2–-N慢性胁迫过程中起重要作用。     

Phytoplankton community structure and diversity in the indoor industrial aquaculture system for Litopenaeus vannamei revealed by high‐throughput sequencing and morphological identification

The explosive multiplication of phytoplankton caused by water eutrophication often occur in the intensive shrimp aquaculture. To comprehensively assess the diversity and community structure of phytoplankton in the waters of typical indoor industrial aquaculture system for Litopenaeus vannamei, a combination of high‐throughput sequencing and morphological identification methods were used in the present study. A total of 41 genera belong to nine phyla were detected by both methods. Chlorophyta, Cyanophyta and Bacillariophyta were found to be three dominant phyla. The high‐throughput sequencing revealed that green algae and cyanobacteria were the most dominant phytoplankton; however, diatoms were the first dominant phytoplankton by using the morphological identification. At the genus level, Picochlorum and Synechococcus were dominant, accounting for 20.94%–97.19% and 0.01%–52.81% of total phytoplankton, revealed by the high‐throughput sequencing. Therefore, more attention should be paid to their ecological impacts on the surrounding sea areas or potential toxicity to shrimp. Cyclotella was the most dominant genus revealed by the morphological identification. High‐throughput sequencing revealed a high diversity and small‐sized phytoplankton which were undetected by microscopy. Both methods provide similar information on the environmental drivers of phytoplankton community. NO₃^?, NH₄⁺, DIP, DSi, DON and DOP concentrations were the main factors influencing the phytoplankton community structure and diversity.     

一株凡纳滨对虾病原菌的分离、鉴定及其致病力分析

从患病凡纳滨对虾肝胰腺中分离出菌株20100612001,经人工感染实验证实,该分离菌株对凡纳滨对虾的半数致死量为1.44×106CFU/ml。形态学观察和革兰氏染色表明,该菌株为革兰氏阴性,无芽孢,有一根端极鞭毛;呈球杆状、球状、棒状或梨状且有两极浓染现象;在2216E培养基上为透明或半透明的圆形菌落,而在TCBS选择性培养基上为绿色或蓝绿色菌落。经Biolog碳源利用反应、脂肪酸气相色谱分析得出,该菌株与副溶血弧菌、需钠弧菌等的生理生化特性最相似;16SrD-NA序列测定表明,该菌株与弧菌属中几株病原菌的同源性均达到98.9%以上。在分子进化树中该菌株与副溶血弧菌Vibrio parahaemolyticus的进化地位最接近。综合上述实验结果分析得出,该细菌分离物为副溶血弧菌。     

Isolation and identification of Vibrio campbellii as a bacterial pathogen for luminous vibriosis of Litopenaeus vannamei

Outbreak of luminescent disease was reported from Litopenaeus vannamei shrimp farms in Zhangpu County, Southern China during May–July 2011. The clinical signs included fluorescent, less food consumption and high mortality. Bacteria were isolated from the infected shrimps. The pathogen, a luminescent bacterium named VH1 was identified as Vibrio campbellii based on MLSA analysis (16S rDNA, rpoD and toxR). The haemolysin (hly) gene specific in V. campbellii was detected in strain VH1. Pathogenicity test using immersion infection confirmed that strain VH1 was virulent to L. vannamei postlarvae and juveniles, and the LC₅₀ value was 1.55 × 10⁶ CFU mL^?1 and 1.7 × 10⁶ CFU mL^?1 respectively.     

Rearing of the Pacific white shrimp,Litopenaeus vannamei in a biofloc system: The effects of different food sources and salinity levels

The present study assessed the effects of different types of feeds and salinity levels on water quality, growth performance, survival rate and body composition of the Pacific white shrimp, Litopenaeus vannamei, juveniles in a biofloc system. Shrimp juveniles (2.56 ± 0.33 g) were cultured for 35 days in 300 L fibreglass tanks (water volume of 180 L) with a density of 1 g/L in six treatments. Three sources of feed (100% formulated feed, mixture of 66.6% formulated diet and 33.3% wet biofloc, and 100% wet biofloc) and two levels of salinity (10 and 32 ppt) were considered in two control groups and four biofloc treatments. Water quality parameters in the biofloc treatments were significantly better than control groups (p < .05). The highest increase in growth performance and survival rate were obtained in salinity of 32 ppt and mixed feed sources. Analysing the proximate composition of body shrimp indicates an increase in lipid and ash levels in biofloc treatments, which was more evident in the salinity of 32 ppt. In addition, the proximate analysis of shrimp body showed significant differences between biofloc treatments and control groups (p < .05). The highest FCR was found in the treatment with salinity level of 10 ppt and fed only with floc. Overall, it was found that the artificial diet supplemented with biofloc at the salinity of 32 showed better performance in the juvenile stage of Pacific white shrimp.     

饲料中添加侧孢短芽孢杆菌FAS05对凡纳滨对虾生长、抗病及免疫力的影响

侧孢短芽孢杆菌(Brevibacillus laterosporu)是一种重要的生物防治益生菌,但在水产养殖中鲜见报道。为研究饲料中添加侧孢短芽孢杆菌FAS05对凡纳滨对虾(Litopenaeus vannamei)生长、抗病及免疫力的影响,以体重为(1.00±0.08) g的凡纳滨对虾为研究对象,开展了28 d的养殖实验。实验分为4组,每组3个重复,每个重复50尾对虾,分别饲喂添加0 (C组,作为对照组)、10⁵ (BL1组)、10⁷ (BL2组)和10⁹ CFU/g(BL3组)侧孢短芽孢杆菌FAS05的实验饲料。结果显示,各组存活率无显著差异(P>0.05),BL1和BL2组对虾的体长、体重和特定生长率显著高于C组(P<0.05);与C组相比,BL1～BL3组养殖水体的弧菌数显著降低(P<0.05);侵染副溶血弧菌(Vibrio parahemolyticus)后,BL1～BL3组存活率显著高于C组(P<0.05),BL1～BL3组之间的差异不显著(P>0.05);与C组相比,BL1～B...     

Genetic parameters and response to selection for harvest body weight of pacific white shrimp,Litopenaeus vannamei

Genetic parameters and response to selection were estimated for the harvest body weight of Litopenaeus vannamei. The data consisted of 24 072 progeny from 178 sires and 171 dams in two generations (G₀ and G₁) with a nested mating structure. All families were randomly divided into two groups and then cultured in two farms at different locations (Huanghua and Qingdao). The heritability estimates from G₀ and G₁ were 0.278 ± 0.136 and 0.423 ± 0.065 respectively. Over two generations, the heritability estimate was 0.335 ± 0.087, and the common environmental effect was 0.084 ± 0.031. A bivariate animal model was used to estimate variance and covariance components, whereby the body weight in the two farms was treated as a genetically distinct trait. Genetic correlation was close to unity (0.943 ± 0.066), indicating that a genotype by environmental interaction for harvest body weight was small. The response to selection in harvest body weight was estimated using two methods (the realized and predicted responses). The realized response was estimated from the difference in the least squares means of body weight for the selection and control populations. The predicted response was obtained from the difference in the mean estimated breeding values between generations. The realized response was 2.30%, while the predicted responses were 2.00% and 1.37% for within‐ and across‐generation datasets using two sets of genetic parameters respectively. The results would provide crucial information in pacific white shrimp breeding programs in China.     

Nursery performance of Pacific white shrimp (Litopenaeus vannamei Boone, 1931) cultivated in a biofloc system: the effect of adding different carbon sources

Effect of different carbon sources on nursery performance of Pacific white shrimp (Litopenaeus vannamei) cultivated in biofloc system was investigated. Shrimp postlarvae (98.47 ± 8.6 mg) were fed for 32 days in tanks with water volume of 130 L and density of 1 individual L^?1. One control treatment and four biofloc treatments (BFT1, BFT2, BFT3 and BFT4) with adding different carbon sources including molasses, starch, wheat flour and mixture of them, respectively, were considered at equal weight ratios. According to the results, salinity, dissolved oxygen and pH were not significantly different among the biofloc treatments (P > 0.05). Maximum pH (8.27) and maximum dissolved oxygen (6.35 mg L^?1) were recorded in the control. Maximum (0.43 mg L^?1) and minimum (0.09 mg L^?1) ammonia were recorded in the control and BFT2, respectively (P < 0.05). Using simple carbohydrates (molasses and starch) lowered the ammonia concentration significantly. The highest increase in body weight (1640.43 ± 231.28 mg), growth rate, specific growth rate (8.97 ± 0.42% per day) and biomass (190.29 ± 26.83 mg) were found in BFT1 and the highest survival (90 ± 0.77%) was found in BFT4. The highest feed conversion (1.52 ± 0.23) and the lowest feed efficiency (66.81 ± 7.95) were observed in the control (P < 0.05). The proximate composition analysis revealed an increase in lipid and ash in biofloc treatments. Results indicated that using biofloc technology with zero‐water exchange system and adding carbon sources could help to recycle waste and improve the water quality. Moreover, the type of carbonaceous organic matter as a substrate for heterotrophic bacteria would be effective in degradation and metabolization of ammonia and nitrite.