生物絮团系统(BFT)中PHBV为碳源对斑点叉尾鮰生长、水质及硝化反应的影响

为了研究生物絮团系统(BFT)中以3-羟基丁酸-co-3-羟基戊酸共聚物(PHBV)为碳源对斑点叉尾鮰(Ictalurus punctatus)生长、养殖水质及对硝化过程反应速率及硝化酶的影响,设置不添加PHBV组(CL组)和挂袋300 g PHBV颗粒组(PHBV组)进行29 d的养殖实验,每4 d检测水质、絮团指标。实验结束检测鱼体生长指标,取实验末养殖水进行10 mg/L NH~+_4降解连续监测,取特征点总氨氮(TAN)最高(A),亚硝酸盐(NO~-_2-N)最高(B_1/B_2)及硝酸盐(NO~-_3-N)最高(C)时的絮团样检测氨单加氧酶(AMO)、羟氨氧化还原酶(HAO)及亚硝酸氧化还原酶(NOR)含量。结果显示:两组的成活率、增重率、特定增长率以及饲料系数均无显著差异,PHBV对斑点叉尾鮰鱼苗生长促进不显著。整个养殖过程中两组TAN、NO~-_2-N均处于安全范围,硝酸盐、磷酸盐、总氮、总磷及总固体悬浮物与絮团体积均呈现不断积累的趋势,且两组将10 mg/L NH~+_4转化为NO~-_3-N均需约28 h,表明两组生物絮团性能良好。硝化酶结果显示,PHBV组的AMO、HAO及NOR含量均显著大于CL组,但其硝化速率并未显著提高。两组游离菌的AMO、HAO、NOR含量均显著低于固着菌,且CL组游离菌中AMO、HAO及NOR含量与PHBV组差异不显著。     

生物絮团系统中的水制备方案:碳源和施肥量对水质和虾性能的影响

正本研究旨在评价不同碳源及其两种施肥量对生物絮团系统中的水质和虾性能的影响。在高密度(150只/m~2)零换水生物絮团技术(BFT)系统中养殖南美白对虾后期幼虫(PL17)。使用12个水池(0.85 m~3/个)进行了为期39 d的试验。试验处理内容包括将米糠(RB)和糖蜜(MO)两个有机碳源各按两种剂量(25 mg/L和100 mg/L的总悬浮固体)添加。在施肥期结束时,RB100和MO100处理组中     

硝化型生物絮团的驯化培养

为培养硝化型生物絮团、减少碳源投加、提高絮团效率并缩短培养周期,文章采用养殖废水排污口底泥为接种污泥培养生物絮团,通过逐渐减少碳源投加,开展了硝化型生物絮团的定向培养,并结合高通量测序分析了生物絮团菌群变化。结果显示,排污口底泥主要优势菌群与其他报道的异养生物絮团一致,具有良好的微生物菌群基础,能够在7 d内形成出水稳定的生物絮团。随着碳源减少,生物絮团微生物菌群结构随之改变,32 d后形成硝化型生物絮团。高通量测序结果显示,接种污泥和硝化型生物絮团主要优势菌群均为变形菌门和拟杆菌门。在纲水平上,原始污泥优势菌群为Gammaproteobacteria (γ-变形杆菌属)、Bacteroidia (拟杆菌属)和Deltaproteobacteria (δ-变形杆菌属),而硝化型生物絮团优势菌群为Bacteroidia、Gammaproteobacteria和Anaerolineae (厌氧绳菌属)。硝化型生物絮团硝化菌总相对丰度对比原始污泥有了较大提高,出水水质稳定,能有效调控养殖后期水质并降低养殖成本。     

不同碳源培养生物絮团对南美白对虾养殖影响试验

以红糖、糖蜜、豆粕作为碳源研究三种碳源培养的生物絮团对南美白对虾养殖的影响,结果表明:生物絮团含量:红糖组>糖蜜组>豆粕组,三种碳源培养的生物絮团能够有效降低水体中的氨氮和亚硝酸盐含量,对pH值影响不大,三个试验组的成活率均高于对照组,其中红糖组>糖蜜组>豆粕组。综上所述:红糖、糖蜜和豆粕作为碳源的生物絮团养殖均能降低水体中的氨氮和亚硝酸盐,提高南美白对虾的成活率,同时生物絮团养殖可以降低换水量,减少养殖废水排放,生物需团技术在今后的南美白对虾及其他经济鱼类养殖过程中值得推广。     

生物絮团对仿刺参幼参消化与免疫酶活性的影响

水温18.2-21.9 ℃和盐度30-32下,在室内200 L塑料水槽中添加不同的碳源（葡萄糖、蔗糖、玉米淀粉、地瓜粉等）形成生物絮团,养殖体重（0.9±0.1 ）g/只的仿刺参（Apostichopus japonicus ）幼参2个月,探讨水体中的生物絮团对其体内主要消化酶和免疫性酶活性的影响。结果表明,养殖水体中添加淀粉、蔗糖有利于提高幼参体内消化酶（淀粉酶、蛋白酶）的活性。复合碳源组（葡萄糖：果糖：蔗糖：玉米淀粉：地瓜粉=0:3:4:2:1）幼参胃蛋白酶活性（10.63 U/mg prot）显著高于其它组,复合碳源更利于提高幼参胃蛋白酶活性。复合碳源组（葡萄糖：果糖：蔗糖：玉米淀粉：地瓜粉=1:2:4:1:2）,幼参体壁中ALP活性最高,为2.66 U/mg;而玉米淀粉组中,幼参体液中SOD活性（204.66 U/mg ）显著高于其他各试验组（P < 0.05）。添加碳源后制得的生物絮团可以提高幼参机体的免疫功能。     

生物絮团在凡纳滨对虾封闭养殖试验中的形成条件及作用效果

本研究尝试将生物絮团技术应用到凡纳滨对虾试验性封闭养殖系统中,筛选生物絮团养殖所需的适宜碳源及其添加量,在此基础上研究生物絮团养殖系统中凡纳滨对虾的适宜养殖密度。结果表明,在养殖密度为150和300尾/m2的凡纳滨对虾养殖系统中,每天按照饲料(蛋白含量42%)投喂量的77%添加蔗糖,生物絮团4d即可形成,在84d的养殖期内,养殖水体的氨氮和亚硝酸氮浓度均维持在较低水平,对虾成活率在80%以上,取得较好的养殖收获。     

无机碳源对生物絮团降氮及沉降性能的影响

利用异位生物絮团反应器,分别在有机碳源存在(第Ⅰ阶段,持续21 d)和有机碳源缺失(第Ⅱ阶段,持续21 d)阶段,比较研究了无机碳源(NaHCO_3)浓度为0.0 (对照组),0.5,1.0和1.5 g/L的模拟养殖废水对反应器生物絮团降氮及沉降性能的影响。结果显示,第Ⅰ阶段对照组出水氨氮浓度显著高于其他处理组,但总体上呈先下降后稳定的趋势,各组亚硝态氮和硝态氮均有少量积累;生物絮团生物量及沉降速度对照组显著低于处理组,处理组之间差异不显著。第Ⅱ阶段各组出水的氨氮、亚硝态氮浓度无显著差异,对照组硝态氮浓度高于各处理组,氨氮浓度迅速下降;此阶段生物絮团的生物量、沉降速度有所下降,NaHCO_3浓度为1.0 g/L处理组表现出较好的沉降效果;粒径分布也趋向均匀。整个实验阶段,不同浓度无机碳源处理条件下,氨氮的去除效率均达到97.8%以上,亚硝态氮无显著积累,处理组生物絮团沉降速度和生物量显著高于对照组。研究表明,添加无机碳源可提高生物絮团降氮性能,增强其沉降速度;移除有机碳源后,生物絮团反应器可维持氨氮去除能力,但引起硝态氮积累,生物絮团生物量减少;有机碳源缺失时,无机碳源(≥0.5 g/L)有助于生物絮团反应器保持其氨氮去除能力。     

基于PCR-DGGE技术分析生物絮团的细菌群落结构

在草鱼养殖过程中添加碳源(葡萄糖)维持水体C∶N为20∶1以培养生物絮团,通过对生物絮团细菌群落构成进行种类鉴定来评价生物絮团中功能微生物的组成.采用PCR-DGGE(变性梯度凝胶电泳)技术分析生物絮团形成第5、10和15天的细菌群落结构.DGGE指纹图谱结果分析表明:第5天和第10天的相似性最高,达67.4％;第5天和第15天相似性系数最低仅为40.5％.第10天时微生物多样性最高,第15天时多样性最低.对DGGE指纹图谱特征条带进行回收、克隆测序,结果表明,生物絮团培养过程主要微生物类群隶属于以下6个纲:α-变形菌纲( Alphaproteobacteria)、γ-变形菌纲(Gammaproteobacteria)、β-变形菌纲( Betaproteobacteria)、放线菌纲(Actinobacteria)、芽孢杆菌纲(Bacilli)、拟杆菌纲(B acteroidetes),其中α-、β-及γ-变形菌占据主要位置.α-proteobacteria为3个阶段的共有优势菌,第5天时特异菌包括食酸菌属(Acidovorax)、气单胞菌属(Aeromonas)、土壤杆菌属(Agrobacterium),第10天和15天分别为芽孢杆菌属(Bacillus)与红球菌属(Rhodococcus).研究首次发现,生物絮团应用于淡水养殖系统时细菌的组成和多样性都极其丰富,通过结合分析这些微生物的功能特点,为生物絮团技术在实际养殖生产中的进一步应用奠定基础.     

不同盐度对生物絮团、对虾生长以及酶活性的影响

在不同盐度条件下进行凡纳滨对虾的生物絮团养殖试验,研究盐度对生物絮团养殖水质和对虾生长及其酶活性的影响。试验设5个盐度梯度(10、15、20、25、30),生物絮团初始量为20 mL/L,对虾密度为500尾/m^3,试验周期30 d。试验结果显示,15盐度组与20盐度组的对虾体质量增长率最大,达70.73%,10盐度组的体质量增长率最小,达50.24%。盐度越高生物絮团生长越快,30盐度组17 d生物絮团沉降量达200 mL/L,之后逐渐降至43 mL/L,其他组呈相同变化趋势。试验过程中水体总碱度与pH持续降低,但不同组间差异不显著(P>0.05)。盐度越高氨氮累积越快,30盐度组在第6 d达到最大质量浓度8.62 mg/L,之后降至0 mg/L,其他组呈相同趋势变化。盐度越低亚硝态氮累积越快,10盐度组在第6 d达到最大质量浓度9.18 mg/L,之后降至0 mg/L,其他组呈相同趋势变化。硝态氮在不同盐度中呈前期上升的趋势,第16 d之后开始缓慢下降。15盐度组的淀粉酶活性显著高于其他组(P<0.05),其他各组之间无显著差异(P>0.05)。脂肪酶在25盐度组活性最高,盐度升高或者降低酶活性均降低。在10、15、20盐度组中,超氧化物歧化酶、碱性磷酸酶、酸性磷酸酶活性均维持在较高水平,在相同盐度下,肌肉酶活性低于肝胰脏。     

饲料源Cu(Ⅱ)在生物絮团水产养殖系统中的积累及其对氨氧化的影响

为提高生物絮团技术的安全性,试验探究硝化型生物絮团水产养殖系统中饲料源铜Cu(Ⅱ)的积累情况和Cu(Ⅱ)对生物絮团氨氧化的影响.通过对生物絮团养殖系统中的水体铜Cu(Ⅱ)和絮团中Cu(Ⅱ)含量进行测定.结果显示:养殖系统中水体Cu(Ⅱ)含量随着饲料投喂而不断上升,在投喂91 d后达到(18.34±0.77)μg/L,絮...     

Intensive hatchery performance of Pacific white shrimp in the biofloc system under three different fertilization levels

Bacterial inorganic nitrogen control using carbon:nitrogen ratio (C:N) manipulation is a tool for aquaculture systems. The present study assessed the hatchery performance of Litopenaeus vannamei between the mysis 1 and postlarvae 5 stages in a zero-exchange biofloc system under different C:N fertilization levels (10:1, 12.5:1 and 15:1) with dextrose. Water quality, performance parameters and water microbiology were compared among treatments. The mean values of the evaluated water quality parameters were appropriate for this production stage. Fertilization with dextrose efficiently controlled ammonia levels, which did not reach the average concentrations considered toxic for the species. In the 10:1 C:N ratio treatment, the levels of ammonia started to increase early and showed significantly higher levels from the third to the last experimental day. There was no difference among groups in means of survival (>76%) and dry weight (0.26 mg) of L. vannamei production parameters and water quality were maintained without water exchange using a biofloc system supplemented with dextrose. Therefore, the use of biofloc systems without water exchange with dextrose as a carbon source in C:N ratios of 10:1, 12.5:1 and 15:1 results in both adequate production indexes and water quality during the misis 1 to post-larvae 5 hatchery phases of L. vannamei. However, the ratios of 12.5:1 and 15:1 keep lower levels of ammonia.     

不同碳源对凡纳滨对虾育苗标粗水体生物絮团的结构、营养成分、细菌群落及其水质的影响

为研究凡纳滨对虾育苗标粗阶段生物絮团形成所需要的适合碳源,设计3种不同碳源添加组(葡萄糖组、淀粉组和蔗糖组),每个处理组设置3个重复,实验期20 d,以分析不同碳源添加后对水体生物絮团的形成、营养成分、细菌群落结构及水质指标的影响。结果显示,在碳源添加量均为投喂量的80%时,形成的生物絮团可有效调节水质,降低水体中的氨氮和亚硝酸盐氮水平。3个碳源添加组水样中氨氮、亚硝酸盐氮和硝酸盐氮浓度显著低于对照组,淀粉组水样中氨氮、亚硝酸盐氮和硝酸盐氮浓度显著高于葡萄糖组和蔗糖组;最终对虾存活率统计结果显示,葡萄糖组、淀粉组、蔗糖组和对照组分别为72.9%、54.2%、69.8%和44.3%;淀粉组的生物絮团沉降体积(BFV)显著低于葡萄糖组,蔗糖组BFV最高,在13～15 d后3组均趋于稳定;葡萄糖组和蔗糖组的粗蛋白含量均显著高于淀粉组,葡萄糖组和蔗糖组则差异不显著;葡萄糖组和蔗糖组生物絮团中组氨酸、精氨酸、蛋氨酸等必需氨基酸和天冬氨酸、谷氨酸、丙氨酸等非必需氨基酸含量都显著高于淀粉组;葡萄糖组、淀粉组和蔗糖组的必需氨基酸指数(EAAI)值分别为0.93、0.89和0.92。3种类型生物絮团在门级水平的细菌群落共有18余种,其中变形菌门和拟杆菌门在各组占有比例均最高,淀粉组拟杆菌门含量显著高于其他2组,蔗糖组浮霉菌门和放线菌门含量显著高于葡萄糖组和淀粉组。研究表明,添加不同碳源可影响水体生物絮团的形成、营养成分、细菌群落结构和多样性,不同程度地改善水质。以必须氨基酸指数及存活率为评价指标,则葡萄糖和蔗糖都是凡纳滨对虾育苗标粗水体中适宜的碳源选择。     

一株异养硝化菌AD-28氨氮去除特性研究

以异养硝化菌——嗜碱假单胞菌AD-28为试验对象,通过单因子试验测定好氧条件下碳源、碳氮比、温度、pH、溶解氧、初始氮质量浓度及盐度等不同因素对嗜碱假单胞菌AD-28生长及氨氮去除的影响。试验结果表明,适用于嗜碱假单胞菌AD-28生长和氨氮去除的最佳条件为:碳源柠檬酸钠、丁二酸钠、乙酸钠,碳氮比20,温度25～35℃,pH 6.0,转速120r/min;在最佳条件下,嗜碱假单胞菌AD-28对初始质量浓度为20～160mg/L的氨氮去除率在24h内达95%以上。嗜碱假单胞菌AD-28能直接以氨氮为底物进行高效的硝化作用,并能耐受较低的温度(15℃)和较高的盐度(NaCl质量浓度为50g/L),在调节养殖水体水质方面具有广阔的应用前景。     

The effect of different alkalinity levels on <Emphasis Type="Italic">Litopenaeus vannamei</Emphasis> reared with biofloc technology (BFT)

The initial stages of rearing marine shrimp using biofloc technology (BFT) involve the biofloc formation process. At the same time, there is an increase in the levels of total suspended solids and a decrease in alkalinity and pH. This reduction of alkalinity and pH occurs due to the consumption of inorganic carbon by the autotrophic bacteria present in the bioflocs and biofilms. The aim of this study was to evaluate the effects of different alkalinities on water quality and the zootechnical performance of the marine shrimp Litopenaeus vannamei in a BFT system. The experiment consisted of four treatments, with three replicates each: 75, 150, 225 and 300 mg CaCO₃/L. To maintain the alkalinity at the established level, sodium bicarbonate was applied. For the experiments, twelve experimental units (area = 0.20 m²) with an effective volume of 50 L were stocked with 30 juvenile L. vannamei (0.20 ± 0.07 g), to achieve a stocking density of 150 shrimps/m² and were maintained for an experimental period of 49 days. The 75 treatment presented the highest levels of ammonia and nitrite throughout the study, compared to the 150 and 300 treatments. The results showed that higher alkalinity favors biofloc formation and the establishment of nitrifying bacteria.     

一株光合细菌的分离筛选及其对无机氮的去除能力

通过对海洋环境污泥进行富集培养及分离筛选得到一株光合细菌,通过16S rDNA全序列分析,结合菌株形态和结构,鉴定其为Ectothiorhodospira magna。研究表明菌株在盐度30‰、28℃、DO 8 mg/L的条件下,对初始浓度分别为280、84、98 mg/L的氨氮、亚硝酸盐氮和硝酸盐氮经过10 d处理的去除率分别为81.83%、46.21%、86.79%。在氨氮、亚硝酸盐氮和硝酸盐氮共存的环境下,菌株首先利用氨氮,之后将亚硝酸盐氮转化成硝酸盐氮。     

Strategies for ammonium and nitrite control in Litopenaeus vannamei nursery systems with bioflocs

In biofloc technology (BFT) rearing systems, nitrogen compounds, specially ammonia and nitrite, have to be controlled by microbial pathways, mainly through the activity of heterotrophic and chemoautotrophic bacteria. The objective of this work was to assess different water preparation strategies (heterotrophic, chemoautotrophic and mature) in BFT system for nursery of Pacific white shrimp (Litopenaeus vannamei). A 35-day study was conducted with post-larvae shrimp (0.08 g) stocked in twelve 300 L tanks at a stocking density of 2000 shrimp m⁻³. The water preparation strategies for shrimp rearing that were evaluated in this study included: i) Heterotrophic treatment, where the water received sugar as a carbon source; ii) Chemoautotroph treatment, where ammonium and nitrite salts were added to the water; and iii) Mature treatment, which was created by the addition of a significant amount of water containing mature biofloc from another established BFT system. In both mature and chemoautotrophic treatments, the nitrification process was able to keep toxic nitrogen compounds (ammonia and nitrite) at low levels without the addition of carbohydrates. In contrast, heterotrophic system showed peaks of ammonia and nitrite during the rearing cycle, and the level of these compounds were found to be higher in this treatment (relative to the mature and chemoautotrophic treatments). The chemoautotrophic system exhibited a lower abundance of bacteria from the family Vibrionaceae in the beginning of the experiment compared to the heterotrophic and mature treatments. The combination of low Vibrionaceae abundance and good water quality resulted in improved growth performance in this treatment. These findings demonstrate the importance of manipulating the environment of BFT systems to induce an enrichment of nitrifying bacteria before stocking shrimp. We have also found that the addition of a carbon source to BFT systems is necessary only in emergency situations, when ammonia spikes need to be controlled.     

盐度调节方式对培养海水养殖生物絮凝系统的影响

在9个容积为100 L的圆柱形养殖水桶中,用鳗鱼饲料为原料培养絮体,比较了3种盐度调节方式培养盐度为30的生物絮凝系统的启动效率。第一组为盐度直接调节组:试验开始时盐度即调为30;第二组为盐度缓慢调节组:闷曝结束后每日盐度增加5,每3 h增加1度;第三组为淡水驯化组:将培养好的淡水生物絮凝系统,按照盐度缓慢调节组的增加方式增加盐度。启动完成后,监测3个处理组对10 mg/L氨氮的去除效果。结果显示,盐度调至30时,淡水驯化组和盐度缓慢调节组系统中氨氮和亚硝酸盐氮含量先于盐度直接调节组降至低水平。培养期间,盐度缓慢调节组和淡水驯化组的絮体沉降性能较盐度直接调节组好。高通量测序分析结果表明,黄杆菌纲是盐度为30的生物絮凝系统中的优势菌纲,鞘脂杆菌纲是盐度直接调节组系统中的优势菌纲,放线菌纲是淡水驯化组中的优势菌纲。Leptobactrrium和norank_f_Segniliparacea是盐度为30的生物絮凝系统的主要优势菌属。启动完成后3个处理组氨氮的去除效果差异不显著(P>0.05),盐度缓慢调节组最有利于海水生物絮凝系统的构建。     

Effects of different carbon sources and dietary protein levels in a biofloc system on growth performance,immune response against white spot syndrome virus infection and cathepsin L gene expression of Litopenaeus vannamei

A 5‐week study was performed to evaluate the effect of spoilage date extract (SDE) as the biofloc carbon source on Litopenaeus vannamei (5.4 ± 0.3 g) performance. The two levels of dietary protein (15% and 25% crude protein) and two carbohydrate sources (molasses‐M and SDE‐P) were tested including: M15, M25, P15 and P25. The minimum (0.2 ± 0.0 mg/L) and the maximum (0.5 ± 0.0 mg/L) of total ammonia nitrogen were observed in the P15 and M25 groups respectively. The highest protein efficiency ratio (6.1 ± 0.3) and protein productive value (112.3 ± 5.8%) were found in the P15 group (p < 0.05). No significant difference was found between biofloc treatments in the expression of cathepsin L gene in hepatopancreas (p > 0.05). The number of total haemocyte count (THC), semigranular cells (SGC) and granular cells (GC) of shrimp in SDE‐based biofloc treatments was relatively higher than those in molasses‐based biofloc treatments. Following the white spot syndrome virus (WSSV) challenge, a significant decrease in THC, SGC, GC and hyaline cell values was observed in all treatments (p = 0.001). Plasma biochemical parameters were significantly influenced by dietary protein levels, biofloc carbon sources as well as WSSV challenge test. In conclusion, SDE successfully could be used as an alternative carbon source for establishing a biofloc system in L. vannamei production.     

Growth performance and robustness of African Catfish Clarias gariepinus (Burchell) in biofloc‐based nursery production with different stocking densities

The aim of this research is to evaluate the effects of biofloc system application at different fish density on the nursery production performance and the robustness of African catfish Clarias gariepinus (Burchell) against Aeromonas hydrophila and salinity stress. An economic analysis was also performed to determine the most optimum fish density in biofloc‐based African catfish nursery production. African catfish with an average body weight of 0.96 ± 0.04 g and average body length of 4.20 ± 0.46 cm, respectively, were distributed in 12 units of circular plastic‐lined tanks (1,020 L). The experiment consisted of four treatments in triplicates, that is, a control without carbon source addition at a density of 4 fish/L, and three biofloc treatments at different densities, that is, BFT4 (4 fish/L), BFT6 (6 fish/L) and BFT8 (8 fish/L). Tapioca flour was used as the organic carbon source in biofloc systems and was added at an estimated C/N ratio of 10. Housing the fish in biofloc systems resulted in higher fish growth, more efficient feed utilization, higher fish robustness against A. hydrophila infection and salinity stress, as well as higher profitability of nursery production. Increasing the fish density resulted in higher mortality. However, higher number of fish produced and lower feed conversion ratio were observed in the treatments with higher density (6 and 8 fish/L). In conclusion, the application of biofloc technology at a density of 8 fish/L could be recommended to increase the production and profitability of African catfish nursery culture.