太湖微囊藻毒素在罗非鱼体内累积及生物降解的初步研究

为了解蓝藻水华期间微囊藻毒素在罗非鱼体内的分布及累积传递过程,2008年6～8月采集了高密度蓝藻池塘及太湖网箱内的鱼样及水样,用ELISA法对鱼样和水样进行微囊藻毒素MC-LR含量的检测.结果表明,池塘水体微囊藻毒素MC-LR含量在0.123～0.514 μg/L,MC-LR含量随着藻密度的下降而降低,对照组水体MC-LR浓度显著高于实验组MC-LR含量.池塘鱼体肌肉组织微囊藻毒素MC-LR累积含量在1.194～3.615ng/g,肝脏组织微囊藻毒素MC-LR累积含量显著高于肌肉组织.将池塘与网箱罗非鱼转至无微囊藻水体中暂养,跟踪检测MC-LR含量变化,池塘和网箱鱼体肌肉组织微囊藻毒素MC-LR含量均低于人体每日可耐受摄入量,而肝脏组织藻毒素MC-LR含量则分别需要经过10～20d自然生物降解后降低至安全摄入量之下.讨论了微囊藻毒素在鱼体组织分布与食物链中的累积传递.     

太湖微囊藻毒素在罗非鱼体内累积及生物降解的初步研究

为了解蓝藻水华期间微囊藻毒素在罗非鱼体内的分布及累积传递过程,2008年6～8月采集了高密度蓝藻池塘及太湖网箱内的鱼样及水样,用ELISA法对鱼样和水样进行微囊藻毒素MC-LR含量的检测。结果表明,池塘水体微囊藻毒素MC-LR含量在0.123～0.514μg/L,MC-LR含量随着藻密度的下降而降低,对照组水体MC-LR浓度显著高于实验组MC-LR含量。池塘鱼体肌肉组织微囊藻毒素MC-LR累积含量在1.194～3.615ng/g,肝脏组织微囊藻毒素MC-LR累积含量显著高于肌肉组织。将池塘与网箱罗非鱼转至无微囊藻水体中暂养,跟踪检测MC-LR含量变化,池塘和网箱鱼体肌肉组织微囊藻毒素MC-LR含量均低于人体每日可耐受摄入量,而肝脏组织藻毒素MC-LR含量则分别需要经过10～20d自然生物降解后降低至安全摄入量之下。讨论了微囊藻毒素在鱼体组织分布与食物链中的累积传递。     

凡纳滨对虾养殖水中藻毒素和环境因子间的相关性

调查了凡纳滨对虾养殖池塘水体中微囊藻毒素的含量及其与环境因子间的关系,为凡纳滨对虾健康养殖提供理论依据.本实验室采用固相萃取(SPE)和高效液相色谱(HPLC)测定其微囊藻毒素MC-LR的含量;研究了MC-LR浓度与氨氮(NH3-N)、总氮(TN)、总磷(TP)和水温等环境因子间的关系.结果发现其水样检测出微囊藻毒素MC-LR的最高浓度为23.15μg/L,氨氮、TN、TP与MC-LR均呈显著性正相关,氨氮、TN、TP、MC-LR与水温均呈显著性负相关.该实验结果验证了在发病的凡纳滨对虾养殖池塘中检出微囊藻毒素MC-LR的现象,初步推论凡纳滨对虾养殖池塘中微囊藻毒素MC-LR的含量与相关环境因子有关.     

尼罗与奥利亚罗非鱼对池塘蓝藻水华及水质影响的研究

利用尼罗罗非鱼和奥利亚罗非鱼进行了池塘抑藻试验，采用血球计数板测定水体中的微囊藻密度，并对水体理化性质进行了检测分析。试验结果表明，罗非鱼能有效消减水体中蓝藻生物量，降低水体藻毒素浓度，并对水体TN、TP产生一定影响。其中奥利亚罗非鱼试验组蓝藻初始密度为1．53-108cells／L，下降至0．86-108eells／L，降低43．79％；尼罗罗非鱼组蓝藻初始密度为1．54-108cells／L，下降至O．51×108cells，L，降低66．88％。水体总磷含量无显著变化，总氮含量有所下降，水体微囊藻毒素MC—LR含量随着蓝藻密度的下降而降低，并讨论了尼罗与奥利亚罗非鱼摄食抑制蓝藻的摄食抑制及其对水质的影响。     

南太湖地区一典型青虾养殖池塘中微囊藻毒素浓度与环境因子间的相关性

采用固相萃取(SPE)和高效液相色谱-串联质谱法(HPLC/MS-MS)对湖州某淡水青虾养殖池塘水体中溶解态微囊藻毒素(MC-LR,MC-RR)浓度进行了监测,研究了微囊藻毒素浓度变化及其与总氮(TN)、总磷(TP)、氨氮(NH3-N)、水温、叶绿素a(Chl-a)等水体环境因子之间的关系。结果显示:养殖水体藻毒素浓度在监测期间波动明显,游离的MC-LR的最高浓度达到了102.3 mg/L,MC-RR的最高值为32.6 mg/L,MCLR的浓度要高于相应时段的MC-RR。MC-LR和MC-RR的浓度变化呈显著性正相关。在整个监测周期内TN、TP与MC-LR均呈显著性正相关,NH3-N、TN与水温均呈显著性负相关,TP与MC-RR呈显著性正相关,Chl-a与TP、MCs呈显著性正相关。     

微囊藻毒素在鲤体内富集的初步研究

为了解微囊藻毒素MC-LR在鲤各组织器官中的生物富集作用,采用腹腔注射法将MC-LR纯品稀释液(0.215μg/g)注射到鲤体内,酶联免疫吸附法(ELISA)检测0、1、3、12、24和48 h时肾脏、肝脏、肌肉、胆囊、空肠和卵巢中MC-LR含量分布和积累规律.结果显示,MC-LR在肾脏含量最高,均值为(1.007±0.120)μg/g(各器官均按干重计),其次是肝脏(0.490±0.060)μg/g、胆囊(0.355 ±0.011) μg/g、空肠(0.210±0.005)μg/g、卵巢(0.082±0.021)μg/g和肌肉(0.047±0.003) μg/g.鲤不同组织器官对MC-LR的富集能力存在较大差异,肾脏是MC-LR的主要靶器官,卵巢中也存在少量MC-LR富集,肌肉对MC-LR的生物富集量远低于其他组织器官.试验48 h时,鲤体内各组织器官中和鱼缸水中MC-LR含量均有一定程度的下降,表明鲤体内对MC-LR有较强的解毒机制.     

不同鱼类和采样时间的养殖池塘环境微生物群落研究

微生物群落是养殖池塘生态系统重要的组成部分。本研究在不同月份采集尼罗罗非鱼和斑点叉尾鮰池塘水体样品,分析水样中的硝酸盐氮、亚硝酸盐氮、氨氮、总氮和总磷等理化指标,利用Biolog-Eco微平板技术分析水体中微生物对各类碳源代谢的平均颜色变化率,利用高通量测序技术分析水体中的菌群结构,以期了解养殖鱼种类和季节对养殖池塘环境及菌群结构的影响。结果表明：冬季淡水养殖池塘水质和菌群结构不同于其他季节,养殖鱼类种类对池塘水体理化指标和水体微生物菌群结构的影响不大。不同采样时间的池塘水体理化指标差异显著,同一采样时间不同养殖鱼类池塘水体理化指标之间无显著差异。其中,1月尼罗罗非鱼养殖池塘水样中的NH4-N含量高于其他月份,显著高于4月和7月的水样(P<0.05)；1月TP含量显著高于1月、4月和7月(P<0.05)。1月斑点叉尾鮰养殖池塘水样中的TP和NO₃-N含量显著高于其他三个月份(P<0.05)。Biolog-ECO微平板技术检测到尼罗罗非鱼和斑点叉尾鮰养殖池塘水样中的微生物群落对碳水化合物和氨基酸的利用能力较强,对酚胺类化合物的利用能力较弱,4月斑点叉尾鮰养殖池塘水体微生物对碳源的利用率高于尼罗罗非鱼养殖池塘。高通量测序结果发现厚壁菌门(Firmicutes)和拟杆菌门(Bacteroidetes)是尼罗罗非鱼和斑点叉尾鮰养殖池塘水体中的绝对优势菌门。hgcI clade、CL500-29 marine group和C39等参与氮磷循环的菌属为淡水池塘养殖水体中的主要菌属。本研究结果为了解淡水养殖环境中微生物群落结构与变化规律、更有针对性的构建淡水养殖池塘尾水生态化处理方法提供科学依据。     

K6Nb10.8O30对微囊藻毒素的光催化降解研究

通过合成含铌类复合氧化物光催化剂,开展其光催化降解藻毒素试验,研究其光催化降解微囊藻毒素的降解效率和机理,为去除水体中微囊藻毒素提供高效环保的水处理技术。采用溶胶凝胶法制备了铌酸盐光催化剂K6Nb10.8O30,利用X射线衍射、扫面电镜、紫外可见漫反射光谱等对光催化剂进行了结构表征,该化合物结构是属于四方相钨青铜结构,空间群为P4/mbm（127）,晶粒呈长方柱性,直径约为150nm,长度约400～600nm,吸收能带为2.92eV;在紫外光（UVA）辐照条件下,考察了催化剂对微囊藻毒素（MC-LR）的光催化降解效果,利用高效液相色谱测定水体中微囊藻毒素浓度。结果表明,光催化剂K6Nb10.8O30能够有效的降解藻毒素,降解过程受到pH值、藻毒素的初始浓度、催化剂投加量和光照的影响;在光辐射强度为556μW/cm2、pH4.24、催化剂投加量为0.375g/L的试验条件下,初始浓度3mg/L的藻毒素在180min之内降解率达到93%;对催化剂反应前后的红外光谱分析表明,藻毒素的降解最终不是被催化剂吸附,而是被光催化降解;动力学研究表明,微囊藻毒素的光催化剂降解反应符合一级反应动力学规律,随着微囊藻毒素初始浓度的增大,反应速率常数逐步降低,半衰期也逐渐变长。     

武汉官桥湖蓝藻毒素BMAA的生物累积与健康风险评估

鉴于蓝藻"水华"能产生新型神经毒素-β-N-甲氨基-L-丙氨酸(β-N-methylamino-L-alanine,BMAA),并通过生物富集作用在水生态系统的各营养级进行传递;选取武汉东湖子湖—官桥湖,采用HPLC-MS/MS分析方法,在蓝藻水华暴发期间,测定湖水、蓝藻及鱼体内游离态和蛋白结合态BMAA毒素的含量。结果表明,在水体中未检测到溶解态BMAA(在检测限0.05μg/L以下),但在微囊藻细胞及鱼体内(干重)均能检测到,含量均值分别为(0.040±0.002)μg/g和(0.32±0.317)μg/g,说明官桥湖在暴发水华后,蓝藻产生了BMAA毒素且被鱼类吸收和放大;不同鱼体(干重)累积BMAA的程度不同,总BMAA含量(干重)分别为鳑鲏(0.243±0.205)μg/g,鲫(0.126±0.040)μg/g,鲤(0.613±0.120)μg/g,鲢(0.028±0.018)μg/g,鳙(0.039±0.021)μg/g,鳊(0.879±0.243)μg/g;鲤和鳊的富集浓度较高,且与其它几种鱼有显著性差异(P0.05)。按照WHO生活饮用水安全标准的建议进行推算,官桥湖鱼肉EDI值(估算的每天摄入量)为1.015μg/(kg·d),显著低于估算出的TDI值(日容许摄入量)0.5mg/(kg·d),基本不会对人造成急性中毒,但由于BMAA毒素为慢性毒素,不能忽视经常食用鱼肉后的累积风险。建议有关部门将东湖水产品的BMAA毒素纳入长期监测。     

贵州百花湖鱼体器官及肌肉组织中重金属的分布特征及其与水体重金属污染水平的相关性

通过对鳃、肝脏、肾脏、心脏、背肌、腹肌和尾肌等器官与肌肉中6种重金属(Pb、Cd、Cu、Cr、As和Hg)的调查, 分析了百花湖野生鲢和鲤体内重金属含量水平、分布特征及其与水体重金属之间的相关性。在所测4种器官和3处肌肉组织中, 鲢和鲤体内重金属平均含量由高到低的顺序为 Cu > Pb > Cr > Cd > As > Hg; 在鲢和鲤各器官中, Cr和Cu表现出较强的相关性, 其相关系数分别为r=0.878(P<0.01)和 r=0.972(P<0.01), 而Cr和Hg只在鲤体内表现出负的相关性 r= -0.782(P<0.05); 以6 种重金属为依托的鱼体器官自身之间的相关性较强, 但各器官与沉积物和水质中重金属平均含量之间的相关性较弱。聚类分析表明, 在两种鱼体内, 这6 种重金属可以分为3类。研究结果表明, Pb主要蓄积在鲤的肝脏和心脏中, Cd主要存在于两种鱼的肾脏中, Cu 主要在肝脏及心脏中蓄积, Cr主要蓄积在两种鱼体的肝脏和心脏中, As在鲤心脏中含量较为突出, 但在鲢各器官中含量差异不大, Hg在整个鱼体器官及组织中的含量较为均匀, 鱼体可食用的肌肉组织中这6种重金属含量普遍较低。沉积物中重金属主要以硫化物形态存在, 生物可利用度低, 可能是鱼体重金属元素平均含量普遍较低的主要原因。     

Biochemical parameters of blood plasma and content of microcystins in tissues of common carp (Cyprinus carpio L.) from a hypertrophic pond with cyanobacterial water bloom

The aim of this study was to evaluate the dynamics of the blood plasma parameters and the content of microcystins in the tissues of common carp ( Cyprinus carpio L.) in relation to the toxic cyanobacterial water bloom. Fish (average body mass 2176±697 g) in the hypertrophic pond were exposed to natural water bloom (dominated by Planktothrix agardhii, Pseudanabaena limnetica and Limnothrix redekei ), which contained microcystins (concentration in biomass 20–181 μg g⁻¹ dry wt, concentration in water 0.3–9.5 μg L⁻¹). Biochemical parameters in fish blood plasma were analysed in 89 fish at 14-day intervals during the whole season (nine sampling periods). Our results demonstrated high variability and fluctuations in the investigated parameters. The content of microcystins and density of cyanobacterial cells correlated with some haematological indices as lipase, alanine–aminotransferase, albumin, magnesium and chlorides. The concentrations of microcystins in the muscle and liver of the fish (determined by high-performance liquid chromatography with mass spectrometer) were below the limit of detection during the monitored period [0.31 ng g⁻¹ fresh weight (f.w.) for the liver and 0.13 ng g⁻¹ f.w. for muscle]. Our study demonstrates that although known cyanobacterial toxin microcystins were not detected in the fish tissues, several biochemical parameters important for the fish physiology were modulated by natural cyanobacterial bloom.     

In situ studies on the bioaccumulation of microcystins in the phytoplanktivorous silver carp (Hypophthalmichthys molitrix) stocked in Lake Taihu with dense toxic Microcystis blooms

The phytoplanktivorous silver carp is an important biomanipulation fish to control cyanobacterial blooms and is also a food fish with the greatest production in China. The accumulation of the hepatotoxic microcystins (MCs) determined by LC-MS in various organs of silver carp was studied monthly in Lake Taihu dominated by toxic Microcystis aeruginosa. Average recoveries of spiked fish samples were 78% for MC-RR and 81% for MC-LR. The highest content of MCs was found in the intestine (97.48 μg g^− 1 DW), followed by liver (6.84 μg g^− 1 DW), kidney (4.88 μg g^− 1 DW) and blood (1.54 μg g^− 1 DW), and the annual mean MC content was in the order of intestine > liver > kidney > blood > muscle > spleen > gallbladder > gill. Silver carp could effectively ingest toxic Microcystis cells (up to 84.4% of total phytoplankton in gut contents), but showed fast growth (from 141 g to 1759 g in 1 year in mean weight). Silver carp accumulated less microcystins in liver than other animals in the same site or other fish from different water bodies at similar level of toxin ingestion. There was possible inhibition of the transportation of the most toxic MC-LR across the gutwall. Muscle of silver carp in Lake Taihu should not be consumed during period of dense Microcystis blooms while viscera were risky for consumption in more months.     

Effect of dietary cyanobacteria on growth and accumulation of microcystins in Nile tilapia (Oreochromis niloticus)

A 12-week growth trial was conducted in a flow-through system to investigate the chronic toxic effect of dietary intake of cyanobacteria on growth, feed utilization and microcystins accumulation in Nile tilapia (Oreochromis niloticus L.) (initial body weight: 5.6 g). Six isonitrogenous and isocaloric diets were formulated to include different contents of cyanobacteria with the dietary microcystins increasing from 0 to 5460.06 ng/g diet. The results showed that dietary intake of cyanobacteria could increase the growth of tilapia while there are no impacts on feed conversion efficiency or mortality. Feeding rate was higher for the diets containing highest cyanobacteria. Microcystins were mostly accumulated in fish liver. The relationship between microcystins contents in muscle, liver, spleen and dietary intake could be described by quadratic equations.Microcystins content in the muscle of Nile tilapia in present study exceeded the upper limit of the tolerable daily intake (TDI) of microcystins suggested by the WHO (0.04 μg/kg body weight/d). It is suggested that Nile tilapia fed on toxic cyanobacteria is not suitable for human food.     

LMG在养殖水体中的残留及其在鱼肌肉中的积累

检测曾投放过孔雀石绿(MG)养殖池塘中水样MG和隐性孔雀石绿(LMG)含量,并以此数据为基础通过室内模拟实验研究了不同浓度LMG养殖水体中生长的罗非鱼(Tilapia)和淡水白鲳(Colussoma brachypomum)肌肉中的积累情况。结果显示:所选取的5个池塘中LMG平均含量为30.32 ng/mL,6个月前使用过孔雀石绿的5个鱼塘的养殖水体中仍有一定量LMG残留。室内模拟实验显示:在LMG初始浓度为1.00 ng/mL的养殖水体中,LMG在罗非鱼和淡水白鲳肌肉中积累浓度分别高达60.0 ng/g和70.8 ng/g;罗非鱼肌肉中脂肪含量为0.53%,淡水白鲳为0.75%,说明脂肪含量高的淡水白鲳更容易积累LMG。同时发现,罗非鱼和淡水白鲳积累LMG浓度与水体中LMG浓度成正相关。     

Microcystin production during algal bloom occurrence in Laguna de Bay, the Philippines

ABSTRACT:   The amount of microcystin in Microcystis aeruginosa bloom was investigated during the rainy season of 1999 in Laguna de Bay, the Philippines. Bloom samples taken from the West Bay and East Cove stations of the lake were studied in relation to the characteristics of environmental conditions. Four types of microcystins, microcystin-LR (MC-LR), microcystin-RR (MC-RR), 6(Z)-Adda-microcystin-RR, and 3-desmethylmicrocystin-LR were identified from the natural bloom samples among which MC-LR was the most dominant type of microcystin. Production of microcystin (88.6 µg/100 mg dried cells) was highest during the first sampling week that coincided with high water transparency and high conductivity. The occurrence of a strong typhoon during the second sampling week had changed the environment drastically, which was characterized by low water transparency, high turbidity, low water temperature, and with trace amounts of MC-LR detected at the East Cove station. Thus, toxin production over time as well as the relationship between Microcystis production and toxin concentration could not be fully evaluated.     

围塘混合养殖环境中重金属在水、生物体和底泥间的富集分布

研究了椒江口围塘混合养殖环境中重金属在水、生物体和底泥间的富集分布,以及该环境中重金属在鲈鱼、鲻鱼、鲶鱼3种鱼样各组织中的分布积累情况。结果表明,与其他海域相比,围塘养殖水体和沉积物中重金属铜的平均含量偏高;3种鱼样中铜的积累规律相同的是皮>鳍>肌肉,镉、锌、铅在鱼体中的相同积累规律是鳍>皮>肉,不同鱼体各组织中重金属的含量为鲻鱼>鲶鱼>鲈鱼,鱼样各组织中重金属的分布积累具有显著性差异。     

Confirmation of catfish, Ictalurus punctatus (Rafinesque), mortality from Microcystis toxins

Unexplained deaths of pond-grown catfish have occurred for many years. At least some of these mortalities could be from cyanobacteria toxins ingested during feeding on floating diets or passively assimilated through gills during breathing. Recently we were able to document algal production and subsequent ingestion of these toxicants by catfish during a mortality event. The causative organism, Microcystis aeruginosa, was the dominant species within the phytoplankton community during the cooler autumn-winter season. Pond conditions included a drop in water temperature by c . 5 °C during the 10 days preceding the fish mortalities. Microcystin-LR, a hepatotoxin produced by Microcystis , was detected in water samples and in catfish liver tissue. Fish exposed to pond water containing this toxic bloom were killed within 24 h. Necropsy of fish revealed congested liver and spleen tissues. The combination of clinical signs, detection of microcystin LR in water and in liver, and death of fish exposed to pond water supports the diagnosis of microcystin toxicosis. More research is needed to identify specific environmental conditions initiating toxin production to model and predict occurrence of these toxic algal blooms.     

饲料脂肪水平对吉富罗非鱼体脂沉积及脂肪酸组成的影响

研究了饲料脂肪水平对吉富罗非鱼(Genetic Improvement of Farmed Tilapia,GIFT)的部分形体指标、肌肉肝脏及腹腔脂肪组织的脂肪沉积及脂肪酸组成的影响,同时探讨了鱼体对饲料脂肪酸的吸收利用能力.实验设置饲料脂肪水平分别为1.73%,3.71%,5.69%,7.67%,9.64%,16.55%共6个梯度组,每组3个重复,饲养90 d.结果显示:5.69%,7.67%及9.64%脂肪组吉富罗非鱼肥满度较高1.73%和16.55%脂肪组的肝体指数显著高于其他实验组(P<0.05);除了1.73%脂肪组,其他各组中饲料脂肪水平越高,鱼体的脏体指数越高.7.67%脂肪组吉富罗非鱼肌肉脂肪含量显著高于3.71%组(P<0.05),同时显著低于16.55%组(P<0.05),但是其肝脏中脂肪含量与其他各组差异均不显著(P>0.05);饲料脂肪水平越高,鱼体的脂肪含量越高,同时不饱和脂肪酸在总脂肪酸中的比例越高.结果表明,饲料脂肪水平影响吉富罗非鱼的部分形体指标,尤其对肝脏形态的影响较为明显.饲料中过多的脂肪容易在肌肉和肝脏组织中沉积,同时鱼体的脂肪含量和脂肪酸组成能够反映饲料的脂肪水平和脂肪酸组成.本研究旨在为饲料脂肪水平的合理设置及脂肪营养对脂肪代谢的调控提供参考.     

Effects of microcystin-containing cyanobacterial extract on hematological and biochemical parameters of common carp (Cyprinus carpio L.)

The aim of the study was to assess the effects of a cyanobacterial extract containing microcystins (MCs) on selected hematological and biochemical parameters in common carp (Cyprinus carpio L.), as well as to determine the accumulation of toxins in fish tissues. The fish were immersed for 5 days in water containing toxins at a final concentration of 12 μg/L of microcystin LR equivalent. Microcystin LR residues were detected in fish liver, reaching 207, 238 and 260 ng/g f.w. of the tissues taken 24 h, 72 h and 5 days after the end of intoxication, respectively. The most substantial changes were found in fish plasma, including increases in creatine kinase, lactate dehydrogenase, ammonia, glucose, aspartate aminotransferase and alanine aminotransferase levels. A decline of about 50% in lysozyme activity was observed by the end of the experimental period. Moreover, a marked increase in ceruloplasmin activity was detected 24 h after the end of intoxication with a subsequent decrease in its activity after 72 h and 5 days. This study concludes that not only consumption of food containing toxins but also MCs dissolved in water may pose a threat to fish health. Additionally, detected changes in lysozyme and ceruloplasmin activity may have distinct effects in fish resistance against pathogens or oxidative stress, which should be taken into account in the future studies.