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.     

三峡水库小江回水区水华期间水环境因子对微囊藻丰度的影响

小江是三峡库区北岸流域面积最大的一条支流，其回水区富营养化备受关注。2016年4月下旬至5月，中旬小江回水区高阳断面和双江断面爆发了水华。以高阳断面和双江断面为监测区域，采用冗余分析方法，探究水华爆发期微囊藻丰度与水环境因子的关系，以期找出水华应急阶段关键水环境因子，为小江回水区微囊藻水华预警监测和防控提供科学依据。结果表明，随着水深逐渐增加，微囊藻的丰度逐渐减少，高阳断面上表层（水下0.5 m）、中层（1/2水深）、底层（底部以上0.5 m）微囊藻丰度为2467.67×104个/L、792.58×104 个/L和729.72×104 个/L；双江断面表、中、底层分别为1337.57×104个/L、298.75×104个/L和108.79×104个/L；高阳断面微囊藻丰度明显高于双江断面。高阳断面的水温、溶解氧和高锰酸盐指数明显高于双江断面，其他水环境指标的差异性不显著。冗余分析Monte Carlo检验结果显示，高阳断面溶解氧（P=0.045）与微囊藻丰度呈显著正相关，电导率（P=0.047）与微囊藻丰度呈显著负相关，双江断面水温（P=0.002）与微囊藻丰度呈极显著正相关，透明度与微囊藻丰度呈显著负相关（P=0.011）。     

Evidence of rapid transfer and bioaccumulation of Microcystin‐LR poses potential risk to freshwater prawn Macrobrachium rosenbergii (de Man)

Microcystins accumulate in aquatic organisms and can be transferred to higher trophic levels, eventually affecting vector animals and consumers. We examined three levels of an aquatic food chain (Microcystis aeruginosa, Daphnia magna and Macrobrachium rosenbergii) to identify the transfer efficiency and risk of microcystin on prawns. Samples were analysed using ultra performance liquid chromatography‐mass spectrometry (MS)/MS and microcystin‐LR (MC‐LR) distributions in prawn tissues were studied. The results showed that prawns accumulate MC‐LR both directly from M. aeruginosa and indirectly through D. magna which was pre‐exposed to M. aeruginosa. MC‐LR was detected in the gills, digestive tracts and hepatopancreas of the prawns 2 h after exposure. MC‐LR accumulated in prawns to 0.49 ± 0.04 μg g^?1 dry weight in hepatopancreas within 24 h, while it was not detected in muscle samples, and rarely appeared in blood samples in such a short period. Although MC‐LR was not detected in muscle, the head including hepatopancreas of the prawns accumulated troublesome amounts of MC‐LR. These results demonstrate that microcystis blooms in prawn farming potentially pose a risk to human consumers, although prawns may be exposed to the bloom for a very short time, hence regular monitoring of blue green algae population is recommended.     

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.     

金鱼藻和狐尾藻对铜绿微囊藻生长及藻毒素释放的影响

通过加入金鱼藻(Ceratophyllum)、狐尾藻(Myriophyllum)与铜绿微囊藻(Microcystis aeruginosa)共同培养的实验方式,测定其过程中藻密度、总氮和总磷、藻毒素含量的变化,研究金鱼藻、狐尾藻对微囊藻生长及藻毒素释放的影响。结果表明,金鱼藻、狐尾藻不仅能很好地抑制铜绿微囊藻的生长,吸收水中的氮磷元素,还能在一定程度上抑制微囊藻毒素的释放。其中狐尾藻对铜绿微囊藻密度的抑制率能高达91.19%,对氮和磷的吸附率分别为66.84%和53.75%,对藻毒素MC-LR的抑制率达到44.23%。     

太湖微囊藻毒素在罗非鱼体内的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月至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含量变化范围在0.123～0.514ug/L间，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间，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10～20天自然生物降解后降低至安全摄入量之下。并讨论了微囊藻毒素在鱼体内的组织分布与食物链中的累积传递。     

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。     

Changes in the nutritional parameters of muscles of the common carp (Cyprinus carpio) and the silver carp (Hypophthalmichthys molitrix) following environmental exposure to cyanobacterial water bloom

The present study evaluated the effect of naturally developing cyanobacteria on the composition of muscles of two commercially important freshwater fish species. Fish were exposed to cyanobacterial biomass including Microcystis aeruginosa and Microcystis ichthyoblabe for 4 weeks. Then, they were transferred to dechlorinated potable water without any cyanobacteria for another 4-week period, thus modelling their preparation for consumers. Samples of muscles were collected every week during exposure and subsequent stay in dechlorinated potable water. The cyanobacterial water bloom of 3.9–6 × 10⁵ cells mL⁻¹ (133–383 μg g⁻¹ of total MC DW) induced statistically significant effects only in the content of fatty acids ( P <0.05; P <0.01) in the common carp ( Cyprinus carpio ), while all studied parameters including the content of dry matter and fat ( P <0.01), proteins ( P <0.05), fatty acid composition ( P <0.05; P <0.01) and some amino acids ( P <0.05) were affected in the silver carp ( Hypophthalmichthys molitrix ). This study has shown that cyanobacteria in the environment of commercially produced fish may decrease the dietetic value of fish muscles.     

三类分子探针对常见赤潮生物的检测

集中使用寡核苷酸、肽核酸和细胞凝素3类探针对来自东海和厦门海域的现场赤潮样品进行了检测,尝试鉴定识别自然水样中有害的赤潮原因种塔玛亚历山大藻,微小原甲藻和纤小裸甲藻,建立和优化了这些探针的检测方法和样品处理程序.结果表明,在东海和厦门海域的赤潮样品中均成功地检出了塔玛亚历山大藻的分布情况,各探针的检测效率为DBA>Tama28S>Tama5S;在东海和厦门海域的赤潮样品中,也成功地检测出了微小原甲藻,各探针的检测效率为:ConA>PM18S02>PM28S02;在厦门海域的赤潮水样中检出了纤小裸甲藻,各探针的检测效率为:WGA>PNATP28S01>TP18S02>TP28S01.各探针检测结果与相关文献的报道吻合较好.比较这3类探针的特异性,其中以PNA探针为最好,其次为DNA;lectin探针的特异性相对较弱.     

浅议养鳗池中的微囊藻

养鳗池中常易引起微囊藻“水华”,国内外一些养鳗技术资料对此藻类的存在和大量繁殖持肯定态度,认为有利于增氧,除氨氮,笔者则认为,微囊藻产生的毒素对鱼类有害,鳗池中培养绿藻（如小球藻等）同样能起到增氧,除氧作用,我国的传统养鱼经验之一,掌握池水的“肥、嫩、爽”也适用于养鳗生产。     

Bottom culture of the sea cucumber Apostichopus japonicus Selenka (Echinodermata: Holothuroidea) in a fish farm,southern China

The bottom culture of southward‐transplanted sea cucumber Apostichopus japonicus in a subtropical fish farm was investigated in a field study at Dapeng Cove, Daya Bay, from January 5–August 5 2011, with the aim of finding the ideal period for culturing A. japonicus in fish farms, and developing an integrated multi‐trophic aquaculture (IMTA) in southern China. Results showed that the bottom‐cultured sea cucumbers survived well (100%) before summer, survival rates decreased to 65.00 ± 21.21% in July, and all animals had died at the end of the study. Specific growth rates of the sea cucumbers were high during winter (1.05 ± 0.03% d^?1), decreased in early spring (0.44 ± 0.11% d^?1) and became negative in the following months. Growth rate was mainly influenced by water temperature, dissolved oxygen and sulphide content; the anoxia caused by water column stratification at the seafloor in the summer were the main causes of mass mortality. Our results indicate that bottom culture in the temperate season (winter and spring, optimally from late November to early April) is a viable way to rear the deposit feeder A. japonicus underneath a subtropical fish farm.     

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.     

Characterization of depth-related changes and site-specific differences of microbial communities in marine sediments using quinone profiles

ABSTRACT:   Respiratory quinone compositions were analyzed by a high-performance liquid chromatography to characterize the depth-related changes and site-specific differences of microbial communities in marine sediments. Two deep-sea sediment samples and one coastal sediment sample were investigated from three sites on the coast of Japan, Sagami Bay, Suruga Bay and Tokyo Bay. Although depth-related changes in microbial community structures were observed, site-specific differences appeared to have greater influence on overall community structures. A variety of quinone homologs was commonly identified at all sampling sites and depths examined, but a few minor quinone fractions, mainly derived from Actinobacteria , were detected only at specific sampling sites. Methylmenaquinone-7 (MMK-7) was the major component throughout the examined depths in Sagami Bay and Tokyo Bay. Most abundant quinone homologs changed with each depth in Suruga Bay. Menaquinone-6 (MK-6) predominated in the 0 to 2-cm layer (19%). Below 0–2 cm, the most abundant homolog in each sampling depth was phylloquinone (K₁; 13%, 4–6 cm), MK-8 (19%, 8–10 cm) and MK-7 (13%, 16–18 cm). The microbial respiratory quinone profiling method shown here, successfully demonstrates the usefulness of this approach to characterize microbial communities in marine sediments.     

Confirmation of yessotoxin production by the dinoflagellate <Emphasis Type="Italic">Protoceratium reticulatum</Emphasis> in Mutsu Bay

ABSTRACT:   Yessotoxin (YTX) is a shellfish toxin and its contamination in bivalves has seriously damaged shellfish industries. The biogenetic origin of YTX was identified as the dinoflagellate Protoceratium reticulatum (Claparède et Lachmann) Bütschli collected in New Zealand and Yamada Bay, Iwate in Japan. Scallops cultured in Mutsu Bay, Japan, were frequently contaminated with YTX, however, occurrence of P. reticulatum in this bay and YTX production by the local strains have not been investigated. Eight strains of P. reticulatum , isolated from the bay, were cultured in the laboratory, and analyzed by fluorometric high-pressure liquid chromatography and liquid chromatography/mass spectrometry methods for YTX production and composition. All strains tested were confirmed to produce YTX, and none of them produced known YTX analogs. Toxin amount and composition differed from strain to strain. This result is also confirmation of one of the biogenetic origins of YTX in Mutsu Bay.     

杀藻剂对不同表型铜绿微囊藻的作用

分别用形态学和分子标记（ITS序列）方法对一株可保持稳定群体状态的微囊藻（Microcystis）进行了鉴定,发现该菌株为蓝藻水华常见种类铜绿微囊藻（M.aeruginosa）。采用紫外线、Cu2＋和放线菌发酵产物3种常见杀藻方式对不同表型微囊藻作用时发现,杀灭群体微囊藻所需要的杀藻剂剂量更大,作用时间更长。结果显示,在环境胁迫条件下,该菌株（群体株）比M.aeruginosa PCC7806（单细胞株）具有更强的环境适应能力,也提示在微囊藻水华的控制中,杀藻剂所用剂量和作用时间也应考虑微囊藻的表型。     

Co-occurrence of white shrimp, Litopenaeus vannamei, mortalities and microcystin toxin in a southeastern USA shrimp facility

Various freshwater and marine algal toxins are known to affect plants, fishes, mammals, and invertebrates. During recent mortality events in Texas white shrimp aquaculture ponds, water and shrimp tissue samples were analyzed for cyanobacterial toxins and found to contain microcystin-LR. Cyanoprokaryota dominated the phytoplankton assemblage in water from the affected pond, particularly Microcystis aeruginosa and Anabaena sp. Water samples from the affected pond also contained high levels of microcystin-LR (45 μg/l), whereas adjacent ponds had a diatom-green algal assemblage and no measurable toxin. Unialgal isolates of M. aeruginosa from the affected pond produced microcystin-LR. Free microcystin-LR concentrations in dead shrimp hepatopancreas determined by HPLC were 55 μg/g total shrimp weight, whereas shrimp hepatopancreas from the adjacent pond without shrimp mortalities had no measurable toxin. Muscle toxin concentration was below 0.1 μg/g.     

Evidence that channel catfish, Ictalurus punctatus (Rafinesque), mortality is not linked to ingestion of the hepatotoxin microcystin-LR

Catfish farms located in the south-eastern USA using brackish (3–5 g NaCl L⁻¹) well water experience sporadic fish kills sometimes with high mortality. An investigation of three catastrophic losses occurring in this region identified no involvement of infectious diseases or traditional water quality problems, including oxygen, ammonia or nitrite. The high mortality and time course of the problem was indicative of exposure to a toxin. Attempts by other workers to explain the cause of this unique syndrome (high chloride associated toxicosis of catfish, HCTC), suggested that the losses might be because of microcystin-producing blooms of Microcystis aeruginosa , but our investigations failed to support this conclusion . We found that (1) the liver histology of catfish experimentally exposed to pure microcystin-LR is very different from that of catfish sampled during outbreaks of HCTC; (2) measurements of microcystin-LR concentrations in the three cases were far lower than the concentration required to kill catfish by experimental immersion; (3) the HCTC toxin appears to have a short half-life, whereas microcystin-LR does not; (4) experimental gavage of catfish with massive amounts of microcystin-LR does not cause the acute mortality typical of HCTC; (5) outbreaks of HCTC appear to be associated with heavy blooms of Anacystis marina , a halophytic cyanobacteria, not with blooms of M. aeruginosa .     

Penaeus monodon (Fabricius) production related to water quality in East Java, Indonesia

On two shrimp farms in East Java, Indonesia, a total of 16 tambaks stocked with Penaeus monodon (Fabricius) were monitored for water quality during one growing season. The water quality parameter values were compared with production characteristics. Ranges of parameter values remained between acceptable reference values. Influences of farm, tambak and time on water quality were significant. Tambak production was explained for 25% by water transparency. Observed water quality parameter values could not be a reason for recently described decreased production. Therefore, other factors should be investigated.     

Temperature requirements for seed germination and seedling growth of Zostera marina from central Japan

ABSTRACT:   The optimal water temperature in seed germination and the upper critical water temperature in seedling growth were determined for Zostera marina collected from Ise Bay, Japan. The relationship between the seed germination rates and seed storage period (0, 30 and 60 days at 0°C) was also examined. The optimal water temperature for seed germination was in the range from 10 to 15°C regardless of the storage periods, in which germination rates ranged from 35 to 57%. Seedlings grown from seed up to 10 cm in total length were cultured for 1 week under various water temperatures to measure their relative growth rates. The optimal water temperature in growth was in the range from 20 to 25°C; relative growth rates ranged from 2.0 to 2.6%. Seedlings could survive up to a water temperature of 28°C, but most seedlings withered at 29 or 30°C. The optimal water temperatures for seed germination and seedling growth were related to the seasonal changes of water temperature at the sampling site. Although seedlings were rarely observed in the field in summer, they can grow at temperatures as high as 28°C. Therefore, Z. marina may extend its distribution as far as where the summer water temperature is lower than 28°C.     

Reproductive cycle of yellow sea bream <Emphasis Type="Italic">Dentex tumifrons</Emphasis> in Wakasa Bay,the Sea of Japan off central Honshu

ABSTRACT:   To determine the spawning season and reproductive cycle of the yellow sea bream, Dentex tumifrons , in Wakasa Bay, in the Sea of Japan off central Honshu, seasonal change in the gonadosomatic index ( GSI ) was examined and histological observations of both ovaries and testes were conducted. The female GSI gradually increased from May, maintained a high level between August and October and then rapidly decreased. The histological observations of the ovary showed that vitellogenesis began from May and developed until September. Spent fish began to be observed in October and all the fish collected in December were spent. There was only one peak in the seasonal change in GSI of males and females in Wakasa Bay. Thus, yellow sea bream spawn only in autumn in Wakasa Bay. Differences in the reproductive cycle and number of spawning seasons were found between the two different latitudinal areas, Wakasa Bay and the East China Sea (low latitudinal area) where the spawning seasons were spring and autumn. An increase in water temperature is an important factor to initiate gonadal maturation (recrudescence) and is terminated by the short day-length in Wakasa Bay.