Comparative study on the gut microbiotas of four economically important Asian carp species

Gut microbiota of four economically important Asian carp species(silver carp, Hypophthalmichthys molitrix; bighead carp,Hypophthalmichthys nobilis; grass carp, Ctenopharyngodon idella; common carp, Cyprinus carpio) were compared using 16 S rRNA gene pyrosequencing. Analysis of more than 590,000 quality-filtered sequences obtained from the foregut, midgut and hindgut of these four carp species revealed high microbial diversity among the samples. The foregut samples of grass carp exhibited more than 1,600 operational taxonomy units(OTUs) and the highest alpha-diversity index, followed by the silver carp foregut and midgut. Proteobacteria, Firmicutes, Bacteroidetes and Fusobacteria were the predominant phyla regardless of fish species or gut type. Pairwise(weighted) UniFrac distance-based permutational multivariate analysis of variance with fish species as a factor produced significant association(P0.01). The gut microbiotas of all four carp species harbored saccharolytic or proteolytic microbes, likely in response to the differences in their feeding habits. In addition, extensive variations were also observed even within the same fish species. Our results indicate that the gut microbiotas of Asian carp depend on the exact species, even when the different species were cohabiting in the same environment. This study provides some new insights into developing commercial fish feeds and improving existing aquaculture strategies.     

草鱼与鲢鱼肥胖基因克隆与序列分析

肥胖基因产物leptin是调节哺乳动物摄食、能量代谢等生命活动的重要细胞因子。 应用RT-PCR和RACE法获得了草鱼(Ctenopharyngodon idellus)和鲢鱼(Hypophthalmichthys molitrix) leptin基因的全长cDNA序列分别为1 096 bp和1 176 bp,编码173和172个氨基酸。氨基酸序列同源性分析表明,草鱼和鲢鱼的leptin序列与其它鲤科鱼类leptin的同源性较高,而与其他鱼类的leptin同源性很低,但所有鱼类的leptin均含有用于形成二硫键的高度保守的半胱氨酸。系统进化树分析显示,草鱼和鲢鱼leptin与其他鱼类leptin聚于一进化分支。应用PCR和Genome Walker方法,进一步获得了草鱼和鲢鱼leptin基因的内含子和5′侧翼区序列。结果表明,获得的草鱼和鲢鱼leptin基因长度分别为2 129 bp和2 192 bp,含有与其他脊椎动物leptin相似的基因结构（含三个外显子和两个内含子）。本研究为深入研究鱼类肥胖基因结构功能关系与鱼类抗肥胖品系定向遗传选育奠定了良好的基础。     

Effects of a filter-feeding fish [silver carp, Hypophthalmichthys molitrix (Val.)] on phyto- and zooplankton in a mesotrophic reservoir: results from an enclosure experiment

SUMMARY 1. Silver carp, Hypophthalmichthys molitrix (Val.), feeds on both phyto- and zooplankton and has been used in lake biomanipulation studies to suppress algal biomass. Because reports on the effects of silver carp on lake food webs have been contradictory, we conducted an enclosure experiment to test how a moderate biomass of the fish (10 g wet weight m⁻³) affects phytoplankton and crustacean zooplankton in a mesotrophic temperate reservoir.
2. Phytoplankton biomass <30 μm and particulate organic carbon (POC) <30 μm were significantly higher in enclosures with silver carp than in enclosures without fish, whereas Secchi depth was lower. Total copepod biomass declined strongly in both treatments during the experiment, but it was significantly higher in fish-free enclosures. Daphnid biomass was also consistently higher in enclosures without fish, although this effect was not significant. However, the presence of fish led to a fast and significant decrease in the size at maturity of Daphnia galeata Sars. Thus, the moderate biomass of silver carp had a stronger negative effect on cladoceran zooplankton than on phytoplankton.
3. Based on these results and those of previous studies, we conclude that silver carp should be used for biomanipulation only if the primary aim is to reduce nuisance blooms of large phytoplankton species (e.g. cyanobacteria) that cannot be effectively controlled by large herbivorous zooplankton. Therefore, stocking of silver carp appears to be most appropriate in tropical lakes that are highly productive and naturally lack large cladoceran zooplankton.     

Structural conservation and food habit-related liver expression of uncoupling protein 2 gene in five major Chinese carps

The full-length cDNA of grass carp (Ctenopharyngodon idellus) and silver carp (Hypophthalmichthys molitrix) uncoupling protein 2 (UCP2) was obtained from liver. The grass carp UCP2 cDNA was determined to be 1152 bp in length with an open reading frame that encodes 310 amino acids. Five introns (Intron 3, 4, 5, 6 and 7) in the translated region, and partial sequence of Intron 2 in the untranslated region of grass carp UCP2 gene were also obtained. Gene structure comparison between grass carp and mammalian (human and mouse) UCP2 gene shows that, the UCP2 gene structure of grass carp is much similar to that of human and mouse. Partial UCP2 cDNA sequences of bighead carp (Aristichthys nobilis) and mud carp (Cirrhinus molitorella), were further determined. Together with the common carp (Cyprinus carpio) UCP2 sequence from GenBank (AJ243486), multiple alignment result shows that the nucleotide and amino acid sequences of the UCP2 gene, were highly conserved among the five major Chinese carps that belong to four subfamilies. Using beta-actin as control, the ratio UCP2/beta-actin mRNA (%) was determined to be 149.4 +/- 15.6 (common carp), 127.4 +/- 22.1(mud carp), 96.7 +/- 12.7 (silver carp), 94.1 +/- 26.8 (bighead carp) and 63.7 +/- 16.2 (grass carp). The relative liver UCP2 expression of the five major Chinese carps, shows a close relationship with their food habit: benthos and detritus-eating fish (common carp and mud carp) > planktivorious fish (silver carp and bighead carp) > herbivorous fish (grass carp). We suggest that liver UCP2 might be important for Chinese carps to detoxify cyanotoxins and bacteria in debris and plankton food.     

氨氮胁迫下不同食性鱼类仔鱼抗氧化和非特异性免疫的差异响应

为探究氨氮胁迫对不同食性鱼类的影响,研究选取不同食性的4种鱼类(鲢Hypophthalmichthys molitrix、草鱼Ctenopharyngodon idella、团头鲂Megalobrama amblycephala和黄颡鱼Pelteobagrus fulvidraco)仔鱼为研究对象,探讨不同浓度氨氮(0、1、2和3 mg/L)短期胁迫(96h)对其生长、抗氧化和非特异性免疫响应的影响及其分子机制。结果显示:(1)氨氮胁迫导致4种仔鱼的体长生长速度呈现剂量依赖性减缓;(2)不同浓度氨氮导致4种仔鱼体内T-AOC、CAT和GPx含量显著降低(P<0.05), 2和3 mg/L氨氮暴露显著降低了草鱼、团头鲂和黄颡鱼仔鱼体内SOD活力(P<0.05),仅检测到黄颡鱼gpx及鲢sod转录水平出现显著性下调(P<0.05);(3)在不同浓度氨氮胁迫下, 4种仔鱼相关免疫基因转录水平均呈现一定的上调,仅鲢il1β转录水平显著下降(P<0.05),相对地草鱼仔鱼LYZ含量显著性下降,黄颡鱼仔鱼LYZ含量在2 mg/L氨氮组显著性上升(P<0.05)。双因素...     

Comparative analysis of gastrin-immunoreactive and argyrophil cells in bony fish larvae

A comparative study was made of enteroendocrine cells in the larvae of bony fishes (Teleostei): carp (Cyprinus carpio L.), big head (Aristichyts nobilis Richardson), silver carp (Hypophthalmichthys molitrix Valenciennes), and atlantic salmon (salmo salar L.). Argyrophil cells demonstrated by the method after Grimelius were found in all examined fish species. They are located in the pyloric appendices, stomach and intestinal mucosa. Gastrin-immunoreactive cells demonstrated by the immunocytochemical method were located in the pyloric part of the stomach in all examined fishes, except for the atlantic salmon in which they were not identified. The first argyrophil cells were identified at the age of about 10 days, while gastrin-immunoreactive cells proved to appear 5-7 days later.     

Feeding behaviour of yearling and older hybrid grass carp

Hybrid grass carp resulting from the cross of a female grass carp, Ctenopharyngodon idella (Val.), and a male bighead carp, Hypophthalmichthys ( Aristichthys ) nobilis Rich., 12–18 months old ( c . 300 mm T.L.) were studied in a two-part experiment to determine feeding preference and total daily consumption fish^-1 on selected species of aquatic plants. Fish were maintained in circular pools with 6840·8 1 of water inside a temperature-controlled greenhouse. Preference tests were conducted at three temperature ranges; 25–28° C, 17–20° C and 12–15° C. Based on the time to complete consumption or the relative quantity consumed, the most preferred plant was Lemna gibba when in combination with six other species. Chara sp., Najas guadalupensis and Potamogeton peciinatus were readily consumed and considered to be of about equal preference. Ceratophyllum demersum and Myriophyllum brasiliense were least preferred. Hybrid grass carp generally consumed as much plant material species^-1 and in the same order of preference at the 12–15°C range as they did at 25–28° C. In the second part, mean daily consumption (g) fish^-1 at 25·7–31·0° C for five plant species tested separately was as follows: Chara sp. 369·8; Lemna gibba 178·2; Najas guadalupensis 172·6; Hydrilla verticillata 106·4 and Ceratophyllum demersum 8·8.     

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.     

Influence of local-scale abiotic and biotic factors on stress and nutrition in invasive silver carp

Community structure and dynamics in aquatic ecosystems are influenced by a variety of abiotic and biotic factors including productivity, species composition, and temperature. These factors may also affect local-scale community resilience to nonnative species invasions. We used habitat characteristics, zooplankton concentrations, fish abundances, and species composition and richness data collected by two fish population monitoring programs to define relationships that influence stress and nutrition in invasive silver carp (Hypophthalmichthys molitrix). We collected blood samples and quantified nutritional (alkaline phosphatase, cholesterol, protein, and triglycerides) and stress metrics (cortisol and glucose) from individuals across three distinct time periods. Nutritional patterns in silver carp were explained by temperature and food resources, indicating elevated feeding in warmer months. Patterns in blood-based stress parameters were most strongly driven by environmental characteristics, elevating with high water temperatures and increased turbidity. Nutrient levels and community richness parameters did not influence the stress or condition of silver carp, likely due to the absence of limiting resources or competition for this species. Together, our results provide insights into the factors that may influence the spread and distribution of silver carp, as well as the characteristics of habitats that could be vulnerable to future silver carp invasion.     

Effect of temperature on proteinase activities of enteral microbiota and intestinal mucosa of fish of different ecological group

Effect of temperature on proteinases activities of enteral microbiota and of intestinal mucosa was studied in five fish species (roach Rutilus rutilus, crucian carp Carassius carassius, common perch Perca fluviatilis, pike-perch Zander lucioperca, and pike Esox lucius) belonging by the nutrition type to different ecological groups. Essential differences of temperature characteristics of proteinases of intestinal mucosa and of enteral microbiota are revealed in fish belonging by the nutrition type to different ecologic groups. The character of the t0-function of proteinases of intestinal mucosa and enteral microbiota by casein and hemoglobin as a rule is different. The highest values of relative proteinases activities for casein in the zone of low temperatures (38 and 45.3 % of the maximal activity) are found at study of proteinases of enteral microbiota in common perch and crucian carp. The latter indicates a significant adaptability of the enteral microbiota proteinases of common perch and crucial carp to functioning at low temperatures.     

Effects of fish introduction on the length of the tail of cryptomonads in mesocosm experiments

Two different morphs of the small cryptomonad, Plagioselmis prolonga var. nordica with a posterior tail, were observed during summer and fall in the hypertrophic lake, Lake Kasumigaura, Japan. The tail shortened in mesocosms stocked with planktivorous silver carp ( Hypophthalmichthys molitrix Val.) and elongated by more than 50% in mesocosms from which silver carp were removed. The density of Plagioselmis cells increased significantly upon fish stocking and decreased upon fish removal. The tail length was negatively correlated with algal abundance and positively correlated with crustacean densities, but there was no correlation with nutrient levels or physical environmental parameters in the mesocosms. The variation in tail length was induced by the presence/absence of fish, but was not related to their density. However, silver carp manipulation strongly affected the density of the majority of zooplankters and, interestingly, there was a strong correlation between zooplankton density and tail-length change in Plagioselmis. We propose a possible herbivore-induced defense mechanism triggered by the top predator, silver carp.     

Effect of total dissolved gas supersaturation on the survival of common carp (Cyprinus carpio) and silver carp (Hypophthalmichthys molitrix)

Flood discharge results in total dissolved gas (TDG) supersaturation downstream of a dam during the flood period. Fish suffer death from gas bubble disease (GBD) caused by TDG supersaturation. Nonetheless, current studies mainly attach importance to the survival of benthic fish affected by TDG supersaturation in the Yangtze River in China. Few studies have attempted to investigate the survival of pelagic fish influenced by TDG supersaturated water and compare the tolerance characteristics to TDG supersaturation between benthic and pelagic fish. To identify the survival of fish species that inhabit the various water layers affected by TDG supersaturation, silver carp (Hypophthalmichthys molitrix) (pelagic fish) and common carp (Cyprinus carpio) (benthic fish) were chosen to conduct an acute exposure experiment of four different TDG supersaturation levels (125%, 130%, 135% and 140%). The findings illustrated that the two fish species both exhibited evident aberrant behaviours of maladjustment in TDG supersaturated water. Obvious GBD symptoms were also found in the test fish. The survival probability of silver carp and common carp decreased with increasing levels of TDG supersaturation. The median survival time (ST₅₀) values of the silver carp exposed to four levels of TDG supersaturated water (125%, 130%, 135% and 140%) were 26.84, 7.96, 5.56 and 3.62 h, respectively, whereas the ST₅₀ values of common carp were 53.50, 26.00, 16.50 and 11.70 h, respectively. When compared with common carp, silver carp had a weaker tolerance to TDG-supersaturated water and were vulnerable to GBD. It shows that levels above 125% are not safe for common carp survival. In terms of the tolerance threshold value, silver carp merits further investigation because it showed lower tolerance to TDG than did common carp.     

论白鲢的食物问题

本文系作者结合自己的实践和国内外的文献资料,对白鲢食谱、食物的选择性和消化性等问题进行探讨,指出必须从新的角度来理解白鲢的食物问题和衡量养鱼池水的肥度。       

Relationship between meteorological conditions and natural reproduction behavior of the four famous Chinese carps

Known as the four famous carps in China, black carp (Mylopharyngodon piceus), grass carp (Ctenopharyngodon idella), silver carp (Hypophthalmichthys molitrix) and bighead carp (Hypophthalmichthys nobilis) are commercially important fish species with a high production in China. To reveal the relationships between meteorological conditions and the natural spawning behavior of these fishes, we considered six meteorological factors (including wind, rainfall, temperature, air pressure, sunshine hours and humidity) and the weather conditions in 80 spawning events. The results showed that the spawning activities were more likely to be activated in consecutive rainy days or days when weather changed drastically, while the spawning activities showed no tendency for the weather type except consecutive rainy days. Our analyses also showed that the average rainfall in the initial spawning days is higher than that in the spawning time windows (the time from the earliest spawning activity to the latest spawning activity), while other meteorological elements remained at more or less the same values in both time periods; spawning activity tended to happen in days when the average air pressure was going down or the average temperature going up, while the other meteorological elements showed no consistent tendency.     

Shift in pike, Esox lucius L., predation pressure following the introduction of ruffe, Gymnocephalus cernuus (L.) to Loch Lomond

Rapid and dramatic change in the fish community of Loch Lomond has resulted from a series of fish introductions in recent years. A comparison of the diet of pike, Esox lucius L., in 1989–1990 with data from 1955–1967, prior to recent introductions, demonstrates a shift in prey choice. In 1955–1967 powan, Coregonus lavaretus (L.), dominated in the diet (57% of prey by number) by 1989–1990 the introduced ruffe, Gymnocephalus cernuus (L.), was the commonest prey species (44% by number). This shift in pike predation to an abundant population of introduced ruffe has consequences for native species. Assuming that the pike population has not increased in response to increased food availability due to the introduction of ruffe, pike predation pressure on native species will be relieved. This is likely to have the greatest effect on powan. Comparison of the predation rate in 1955–1967 with 1989–1990 supports the hypothesis that the rate of predation on powan has declined, although the effect that this may have on the powan population is unclear, as the role of predation in the regulation of population size is unknown for this species.     

Development of fish communities in lakes after biomanipulation

Biomanipulation measures in the Netherlands are usually a combination of a drastic fish stock reduction and an introduction of pike fingerlings. In three small shallow lakes (Noorddiep, Bleiswijkse Zoom and Zwemlust) these measures resulted in a clear water state and the development of macrophytes. After the measures the fish community developed differently because of the new physical and biological conditions. Results of lake Noorddiep and lake Bleiswijkse Zoom showed that the fish community became more divers. Bream and carp became less dominant and were partly replaced by roach and perch. The importance of the main predator pike-perch was strongly reduced and replaced by pike and perch. The share of piscivorous fish in the total fish stock increased at all sites. The recruitment of young-of-the-year was similar or even higher in the clear overgrown areas than in the turbid water before the measures, but the recruitment of young-of-the-year to older fish differed between the species. Predation by pike and perch could not control the young-of-the-year cyprinids, but their predation may have contributed to the shift from bream to roach, because of selective predation on bream in the open water, while roach was hiding in the vegetation. The macrophytes provide new refugia and feeding conditions that favour roach and perch, but offer relatively poor survival conditions for bream and carp.     

长江水系钱草鱼遗传结构及变异性的RAPD研究

采用随机扩增多态性DNA（RAPD）技术对长江中游的湖北嘉鱼与江西瑞昌两个地理群体和中游的汉江和湘江两大水系的鲢和草鱼的群体进行了遗传学研究。发现长江水系链的遗传变异要高于草鱼,与现今生物量成反比的反常现象。鲢遗传多样性从大到小的分布是嘉鱼→湘江→瑞昌→汉江。草鱼遗传多样性从大到小的分布是瑞昌→汉江→湘江→嘉鱼,鲢和草鱼的遗传多样性地理分布并不一致。遗传分化指数Gst分别为12.3%和17.5%,表明鲢和草鱼的四个地理群体的遗传分化度较低,可能与地理较近和基因交流频繁有关。     

Amylolytic activity in fish intestinal mucosa: temperature effects

The activity, temperature characteristics and energy of activation of amylolytic enzymes in the intestinal mucosa were studied in six species of fish living in a boreal zone [burbot (Lota lota L.), northern pike (Exos lucius L.), perch (Perca fluviatilis L.), bream (Abramis brama L.), roach (Rutilis rutilis L.), and carp (Cyprinus carpio L.)] and in three species from tropical and subtropical areas [pilchard (Sardina pilchardus W.), jack mackerel (Trachurus trecae C.) and round sardinella (Sardinella aurita V.)]. The amylolytic activity correlated with the feeding habits: it was essentially lower in predators. The enzyme activity at low temperature, relative to the maximal activity, was correlated with the natural environmental temperature where the species lived. At low temperature the relative activity was higher in boreal fish than in tropical and subtropical fish. We found a breakpoint in the Arrhenius plots in all fish species, except for jack mackerel. The energy of activation in predators decreased below the breakpoint in the low-temperature region. The energy of activation in benthophages of the Aral-Ponto-Caspian area was lower at higher temperatures above the breakpoint. A reduction in activation energy in the range of physiological temperatures might indicate adaptation to the environmental temperature.     

武汉东湖鲢、鳙生长的几个问题的研究

采用退算法研究东湖鲢、鳙的生长,包括生长的基本模式,瞬时生长率及其变动,雌、雄鱼生长的差异等问题:1)鲢、鳙的生长模式可用von Bertalanffy方程来描述,其中鲢的参数l∞=99.8厘米、W∞=17,104克、K=0.3040、t0=0.4821年,鳙的参数l∞=117.6厘米、W∞=29,311克、K=0.3088、t0=0.5392年;2)鲢、鳙的瞬时生长率随着年龄的增长而逐渐下降,并且不同世代同龄鱼的瞬时生长率有一定的变动,其变差系数与年龄呈正相关;3)鲢、鳙的生长无显著的性别差异。通过对研究结果的综合分析,阐述了鲢、鳙4龄前后出现生长的拐折主要由其个体大小所制约,种群的不同世代和同一世代的不同个体一样存在生长补偿作用,以及瞬时生长率与肥满系数之间有着明显的对应关系。所取得的有关参数值可以应用于渔业估算。