Molecular cloning and characterization of alpha-class glutathione S-transferase gene from the liver of silver carp, bighead carp, and other major Chinese freshwater fishes

Two full-length cDNAs encoding glutathione S-transferase (GST) were cloned and sequenced from the hepatopancreas of planktivorous silver carp (Hypophthalmichthys molitrix) and bighead carp (Aristichthys nobilis). The silver carp and bighead carp GST cDNA were 920 and 978 bp in length, respectively, and both contained an open reading frame that encoding 223 amino acids. Partial GST cDNA sequences were also obtained from the liver of grass carp (Ctenopharyngodon idellus), crucian carp (Carassius auratu), mud carp (Cirrhinus molitorella), and tilapia (Oreochromis nilotica). All these GSTs could be classified as alpha-class GSTs on the basis of their amino acid sequence identity with other species. The three-dimensional structure of the silver carp GST was predicted using a computer program, and was found to fit the classical two-domain GST structure. Using the genome walker method, a 875-bp 5'-flanking region of the silver carp GST gene was obtained, and several lipopolysaccharide (LPS) response elements were identified in the promoter region of the phytoplanktivorous fish GST gene, indicating that the GST gene expression of this fish might be regulated by LPS, released from the toxic blue-green algae producing microcystins. To compare the constitutive expression level of the liver GST gene among the six freshwater fishes with completely different tolerance to microcystins, beta-actin was used as control and the ratio GST/beta-actin mRNA (%) was determined as 130.7 +/- 6.6 (grass carp), 103.1 +/- 8.9 (bighead carp), 92.6 +/- 15.0 (crucian carp), 72.3 +/- 7.8 (mud carp), 58.8 +/- 11.5 (silver carp), and 33.6 +/- 13.7 (tilapia). The constitutive expression level of the liver GST gene clearly shows that all the six freshwater fishes had a negative relationship with their tolerance to microcystins: high-resistant fishes (phytoplanktivorous silver carp and tilapia) had the lowest tolerance to microcystins and the high-sensitive fish (herbivorous grass carp) had the highest tolerance to microcystins. Taken together with the reciprocal relationship of constitutive and inducible liver GST expression level in some of the tested fish species to microcystin exposure, a molecular mechanism for different microcystin detoxification abilities of the warm freshwater fishes was discussed.     

Molecular characterization of glutathione peroxidase gene from the liver of silver carp, bighead carp and grass carp

The cDNAs encoding glutathione peroxidase (GPx) were cloned and sequenced from the liver of three Chinese carps with different tolerance to hepatotoxic microcystins, phytoplanktivorous silver carp (Hypophthalmichthys molitrix) and bighead carp (Aristichthys nobilis), and herbivorous grass carp (Ctenopharyngodon idellus). Using genome walker method, a 750 bp 5'-flanking region of the silver carp GPx gene was obtained, and several potential regulatory elements were identified in the promoter region of the GPx gene. The silver carp GPx gene was widely expressed in all tissues examined. Despite phylogenetic analysis, assigning this newly described carp GPx to the group of mammalian GPx2, the carp GPx seems more similar to GPx1 from a physiological point of view. The constitutive expression pattern of the three carp liver GPx gene, shows a positive relationship with their tolerance to microcystins.     

淡水鱼类微囊藻毒素去毒酶基因的克隆

微囊藻毒素去毒酶在鱼类微囊藻毒素去毒过程中起着关键作用,研究成功克隆鲢鱼、鳙鱼、草鱼、鲫鱼、鳜鱼、罗非鱼等淡水鱼类微囊藻毒素去毒酶基因cDNA核心片段而首次获得这些淡水鱼类微囊藻毒素去毒酶氨基酸序列。鲢鱼、鳙鱼、草鱼、鲫鱼、鳜鱼、罗非鱼微囊藻毒素去毒酶基因与人、小鼠、大鼠、牛、猪、羊的谷胱甘肽S-转移酶基因氨基酸同源性为60%左右,表明淡水鱼类微囊藻毒素去毒酶基因在进化上变异性较大,与其承担微囊藻毒素去毒代谢之特殊功能相适应。     

老江河故道四大家鱼天然资源库研究

扼要介绍了老江河故道四大家鱼天然资源库的结构,并根据老江河故道水域生态系统的结构和功能特点,提出了四大家鱼的放养量应在重视种质质量的基础上,合理调控其放养结构和密度,此外文中还就保护和合理利用长江四大家鱼种质资源的有效途径以及维持老江河四大家鱼天然资源库健康运行的关键进行了分析探讨。     

Introduction and cage culture of exotic carps and their impact on fish harvested in Lake Begnas,Nepal

The suitability of exotic carps namely Aristichthys nobilis (Bighead carp), Hypophthalmichthys molitrix (Silver carp), Ctenopharyngodon idellus (Grass carp), Cyprinus carpio (Common carp) and Labeo rohita (Rohu) in a sub-tropical lake was evaluated. The impact of their introduction on native fishes was also studied. After the introduction and cage culture of exotic carps the total harvest reached 92 kg·ha^–1; an increase of 266% within eight years. The planktivorous bighead and silver carps were most successful. The harvest of the other three species was poor. Since the introduction of exotic carp the harvest of indigenous fishes declined by 42%. Considering the food habits of these fish, further stocking should be limited to bighead and silver carps to limit the adverse effects on the indigenous species.     

In situ study on effect of food competition on diet shifts and growth of silver and bighead carps in large biomanipulation fish pens in Meiliang Bay, Lake Taihu

Silver and bighead carps were cultured in large fish pens to reduce the risks of cyanobacterial bloom outbreaks in Meiliang Bay, Lake Tauhu in 2004 and 2005. Diet compositions and growth rates of the carps were studied from April to November each year. Both carp species fed mainly on zooplankton (>50% in diet) in 2004 when competition was low, but selected more phytoplankton in 2005 when competition was high. Silver carp had a broader diet breadth than did bighead carp. Higher densities and fewer food resources increased diet breadths but decreased the diet overlap in both types of carps. It can be predicted that silver and bighead carps would be released from diet competition and shift to feed mainly on zooplankton at low densities, decreasing the efficiency of controlling cyanobacterial blooms. Conclusively, when silver and bighead carps are used to control cyanobacterial blooms, a sufficiently high stocking density is very important for a successful practice.     

Reproductive condition and occurrence of intersex in bighead carp and silver carp in the Missouri River

Little is known about the reproductive biology of the exotic bighead carp Hypophthalmichthys nobilis and silver carp Hypophthalmichthys molitrix in the Missouri River. In order to fill this gap in understanding, herein is described the reproductive condition of these Asian carps. Evidence is presented which indicates that bighead and silver carp in the Missouri River have a protracted spawning period that extends from early spring through fall and some individual bighead and silver carp are spawning multiple times during a reproductive season. Although bighead and silver carps are successfully maturing and spawning in the Missouri River some reproductive abnormalities such as intersex, atresia, and sterility were observed. Knowledge of the reproductive activity of these invasive carps may be useful to resource managers tasked with their control. Furthermore, the reproductive abnormalities observed should be considered when evaluating the environmental condition of the Missouri River relative to supporting a healthy fish fauna.     

葛洲坝水利枢纽兴建后对青、草、鲢、鳙繁殖生态效应的研究

本文分析了葛洲坝枢纽截流后,大坝上、下游草、青、鲢、鳙的繁殖条件,产卵场位置,产卵规模,鱼卵、鱼苗成色以及4种鱼的群体组成等。指出除原宜昌产卵场发生变化外,其余产卵场基本存在,并在上游新发现了几处产卵场。讨论了大坝对4种鱼的影响程度和上游存在草鱼地方性群体,论证了4种鱼不必过坝产卵的理由。       

Long-term changes in fish community structure in relation to the establishment of Asian carps in a large floodplain river

The Upper Mississippi River System, including the Illinois River, has been invaded by a number of nonnative species including silver carp Hypophthalmichthys molitirx and bighead carp H. nobilis, collectively referred to here as Asian carps. Silver carp densities in the Illinois River have increased dramatically and now represent some of the highest densities of wild silver carp anywhere in the world. Asian carps have the potential to alter existing ecosystems by consuming planktonic resources and therefore, could have the ability to alter existing fish communities as most fishes are dependent on planktonic resources during early development. However, identifying the relationship of fish community structure to the establishment of Asian carps has yet to be thoroughly investigated. Using long-term fish community data collected by the Upper Mississippi River Restoration’s Long Term Resource Monitoring element, we investigate changes in fish community structure pre- and post-establishment of Asian carps. Significant differences in the pre- and post-establishment communities were observed for the majority of gears and habitats. Species contributing to changes between establishment periods included most sportfish species and catostomids, which were less abundant post-establishment of Asian carps, while shortnose gar, grass carp, and emerald shiner were more abundant. While our analyses show Asian carps are likely contributing to major differences in fish community structure, future research and long-term monitoring should investigate the mechanisms and interactions responsible for community changes as well as identifying any potential concurrent or confounding factors such as changes in river hydrology or sedimentation.     

Molecular characterization of heat shock protein 70 genes in the liver of three warm freshwater fishes with differential tolerance to microcystin-LR

Heat shock protein 70 (HSP70) protect cell from oxidative stress by preventing the irreversible loss of vital proteins and facilitating their subsequent regeneration. Silver carp (Hypophthalmichthys molitrix), grass carp (Ctenopharyngodon idellus), and Nile tilapia (Oreochromis nilotica) are three warm freshwater fishes with differential tolerance to microcystin-LR (MC-LR). Full-length cDNAs encoding the HSP70 were cloned from the livers of the three fishes. The HSP70 cDNAs of silver carp, grass carp, and Nile tilapia were 2356, 2348, and 2242 bp in length and contained an open-reading frame of 1950 bp (encoding a polypeptide of 649 amino acids), 1950 bp (649 amino acids), and 1917 bp (638 amino acids), respectively. Like mammalian HSP70, the HSP70 of the three fish was also composed of an ATPase domain from residues 1 to 383 (44 kDa), substrate peptide binding domain from residues 384 to 544 (18 kDa), and a C-terminus domain from residues 545 to 649 (10 kDa). The relatively high conservation of HSP70 sequences among different vertebrates is consistent with their important role in fundamental cellular processes. Using beta-actin as an external control, RT-PCR within the exponential phase was conducted to determine the constitutive and inducible expression level of HSP70 gene among the three fishes (6-12 g) intraperitoneally injected with MC-LR (50 μg kg(-1) body weight). Both constitutive and inducible liver mRNA levels of the fish HSP70 genes showed positive relationships with their tolerance to MC-LR: highest in Nile tilapia, followed by silver carp, and lowest in grass carp. The differential expression pattern of liver HSP70 genes in the three fish indicated a potential role of HSP70 in the detoxification process of MC-LR.     

Abundant and constant expression of uncoupling protein 2 in the liver of red sea bream Pagrus major

Four overlapping cDNA fragments encoding a partial sequence for uncoupling protein 2 (UCP2) were amplified by PCR using degenerate primers from the liver of a marine teleost fish, red sea bream (Pagrus major). The partial sequence was 674 bp long, encoding 224 amino acids. The deduced amino acid sequence from the cDNA partial sequence contained the signature motifs for mitochondrial transporter protein and revealed positional identity higher than 72.8% with UCP2 from mammals. The fish UCP2 gene was highly expressed in the liver but almost undetectable in the visceral mesenteric adipose tissue. Using beta-actin as control, the UCP2 mRNA level was determined to be at least 20-fold higher in the liver than in the visceral mesenteric adipose tissues. Neither 48 h starvation nor high lipid diet had any significant effect on liver UCP2 gene expression, indicating that the abundant UCP2 gene expression was stable and might have some basic function in a fish liver that always contains high lipid content. The striking contrast of UCP2 gene expression in the two fish fat-depot organs is consistent with their large differences in oxidative capacity. We suggest that the fish liver may adapt to a constantly high fat deposit by maintaining high UCP2 expression to constrain reactive oxygen species (ROS) production and protect hepatocytes from apoptosis.     

Changes in abundance of larvae of the four domestic Chinese carps in the middle reach of the Yangtze River, China, before and after closing of the Three Gorges Dam

The four domestic Chinese carps, i.e. grass carp (Ctenopharyngodon idella), black carp (Mylopharyngodon piceus), silver carp (Hypophthalmichthys molitrix) and bighead carp (Hypophthalmichthys nobilis), cast their drifting eggs in the running water of the Yangtze River. In 1997 ~ 2005, a systematic study on the dynamics of the abundance the larvae of the four domestic Chinese carps was performed in the middle reach of the Yangtze River, close to Sanzhou city. It was calculated that in breeding seasons from early May to late June every year the abundance of fry drifting through the section of Sanzhou was 3.59 billion in 1997, 2.75 billion in 1998, 2.15 billion in 1999, 2.85 billion in 2000, 1.90 billion in 2001, 1.90 billion in 2002, 406 million in 2003, 339 million in 2004 and 105 million in 2005 respectively. Several environmental parameters related to the dynamics of larval abundance were also examined simultaneously. The construction of the Three Gorges Dam near Yichang between the upper and middle reaches of the Yangtze River has apparently had a drastic influence on the abundance of fish larvae.     

Development of microsatellite DNA markers of silver carp (Hypophthalmichthys molitrix) and their cross‐species application in bighead carp (Aristichthys nobilis)

Forty‐four microsatellite DNA markers were developed for silver carp, and used to investigate polymorphisms of 41 wild silver carps and seven wild bighead carps collected from Jingzhou fragment of Yangtze River. In silver carp, 40 markers were polymorphic. A total of 297 alleles were detected at 40 polymorphic loci. The number of alleles per locus varied from two to 16 with an average of 7.4 and the expected heterozygosities of each locus ranged from 0.07 to 0.91 with an average of 0.69. All markers amplified both silver carp and bighead carp DNAs.     

Persistence of DNA in Carcasses,Slime and Avian Feces May Affect Interpretation of Environmental DNA Data

The prevention of non-indigenous aquatic invasive species spreading into new areas is a goal of many resource managers. New techniques have been developed to survey for species that are difficult to capture with conventional gears that involve the detection of their DNA in water samples (eDNA). This technique is currently used to track the invasion of bigheaded carps (silver carp and bighead carp; Hypophthalmichthys molitrix and H. nobilis) in the Chicago Area Waterway System and Upper Mississippi River. In both systems DNA has been detected from silver carp without the capture of a live fish, which has led to some uncertainty about the source of the DNA. The potential contribution to eDNA by vectors and fomites has not been explored. Because barges move from areas with a high abundance of bigheaded carps to areas monitored for the potential presence of silver carp, we used juvenile silver carp to simulate the barge transport of dead bigheaded carp carcasses, slime residue, and predator feces to determine the potential of these sources to supply DNA to uninhabited waters where it could be detected and misinterpreted as indicative of the presence of live bigheaded carp. Our results indicate that all three vectors are feasible sources of detectable eDNA for at least one month after their deposition. This suggests that current monitoring programs must consider alternative vectors of DNA in the environment and consider alternative strategies to minimize the detection of DNA not directly released from live bigheaded carps.     

Study of the haemoglobin content of a single erythrocyte (M index) in various cultured fish species

Investigations of the erythrocyte count, haemoglobin content and M index of 7 fish species were performed under equivalent environmental conditions in the same period of the year. The species were: bighead Hypophthalmichthys nobilis (Richardson), carp Cyprinus carpio L., silver carp Hypophthalmichthys molitrix (Val.), grass carp Ctenopharyngodon idella (Val.), sheat-fish Silurus glanis L. , pike Esox Indus L., pike-perch Lucioperca lucioperca (L.). Bighead and carp are carnivorous and omnivorous, silver carp and grass carp are herbivorous and sheat-fish, pike and pike-perch are predatory. Significant differences were demonstrable between M values of fish living on different type of food, where the M index is: It is concluded that M index values are useful as one parameter differentiating between fish with distinctly different food habits.     

鳙鱼同工酶发育遗传学研究

采用淀粉或聚丙烯酰胺凝胶电泳法分析鳙鱼早期发育阶段(从未受精卵到卵黄吸尽期)及成体不同组织(脑、眼、心、肌、肾、肝)中六种同工酶(LDH,MDH,IDH,ADH,SDH,EST)的分化表达模式。鳙鱼同工酶基因的表达具有明显的组织特异性。早期发育阶段,ADH和SDH均无染色活性;LDH、MDH和IDH具有不同的发育变化谱式,而EST酶谱在整个早期发育阶段均无明显变化。与鲢、草鱼相比,鳙鱼早期发育过程中胚胎Ldh-A基因激活的时间被推迟。上述结果可为鳙鱼种群的生化遗传结构分析以及鳙鱼的人工育种提供基础资料。     

Tag retention and survival of juvenile bighead carp implanted with a dummy acoustic tag at three temperatures

Bighead carp Hypophthalmichthys nobilis and silver carp Hypophthalmichthys molitrix (together, the bigheaded carps) are invasive fishes in North America that have resulted in substantial negative effects on native fish communities and aquatic ecosystems. Movement and behavior of adult bigheaded carps has been studied previously using telemetry, while similar studies with juvenile bigheaded carps have yet to be attempted. Recent technological advances in telemetry transmitters has increased the availability of tags sufficiently small enough to implant in juvenile carps. However, the effects of surgical implantation of telemetry tags on juvenile bigheaded carps have not been evaluated. We determined tag retention and survival associated with surgical implantation of acoustic telemetry tags into juvenile bighead carp (range 128–152 mm total length) at three temperatures (13, 18, and 23°C). In addition, we assessed the effect of surgically implanted transmitters on the fitness, defined as changes in weight or critical swimming speed, of carp implanted with transmitters. Survival was high among tagged fish (85%) with 47% of tags retained at the conclusion of the 45‐day study. No substantial decline in fitness of the fish was observed in tagged fish compared to untagged fish.     

用鲢鳙直接控制微囊藻水华的围隔试验和湖泊实践

为了控制水体富营养化所形成的水华,国外经典的生物操纵论者提倡放养食鱼性鱼类以控制食浮游生物的鱼类,藉此壮大浮游动物种群,然后借助浮游动物遏制藻类.按照这条思路,以浮游生物为食的鲢和鳙应该是清除的对象.本文作者认为我国湖泊中危害性最大的是微囊藻水华,而浮游动物根本不能摄食这种水华,不如直接利用鲢鳙进行控制,因而1989-1992年间在武汉的东湖进行了三次原位围隔试验,2000年再度重复了一次围隔试验,结果证明,迄今在东湖中已消失18年的微囊藻水华,重新出现在不养鱼的围隔里.在养入一定密度的鲢或/和鳙的围隔中,就没有水华出现.已经出现水华的围隔,放入适当密度的鲢或/和鳙后,在短期内水华又复消失;而放入草鱼的围隔,则水华始终持续,不受影响.说明鲢和鳙能有效地遏制微囊藻水华.鲢、鳙遏制水华的有效放养密度(亦即有效生物量)为46-50g·m^-3.东湖湖泊中鲢、鳙的生物量,直到1985年才达到这个水平,所以1985年湖里反常地没有出现水华.往后的年份,东湖的鱼产量越来越高,也就是湖里鲢、鳙的生物量越来越大,所以迄今没有微囊藻卷土重来的机会.东湖的水质和其他非生物条件仍适于蓝藻水华的发生与发展,如果东湖鲢、鳙的产量下降到阈值以下,亦即降到1985年以前1000t水平,那么微囊藻及其它蓝藻水华的重来将是不可避免的.经典的生物操纵论学说和措施不能解决微囊藻水华猖獗的问题.相反,用滤食浮游生物的鲢和鳙直接遏制微囊藻却是有实验根据并已被渔业生产实践证明行之有效的.     

长江,珠江,黑龙江鲢,鳙和草鱼原种种群形态差异

对长江、珠江、黑龙江的鲢、鳙、草鱼原种种群的十项形态特征,于高维空间上用统计方法进行判别分析,发现种群间具有显著差异,不同江河鲢、鳙的侧线鳞数也有明显不同。这些形态特征上的差异的大小与种群间的地理距离呈正相关。 根据上述显著性差异的特征,用模式分类法推导得鉴别程序,可在IBM-PC-AT计算机上对鱼逐尾自动判别鉴定其种群,除长江鲢外,准确率在75％～100％。