Sixteen polymorphic microsatellites in bighead carp (Aristichthys nobilis) and cross-amplification in silver carp (Hypophthalmichthys molitrix)

A (GT)(n) enriched partial genomic library of bighead carp (Aristichthys nobilis) was constructed by employing the (fast isolation by AFLP of sequences containing repeats) FIASCO protocol. Sixteen loci exhibited polymorphism with two to seven alleles/locus (mean 3.263) in a test population and the observed heterozygosity ranging from 0.100 to 0.690 (mean 0.392). Eleven of the 16 bighead carp microsatellites were found to be also polymorphic in silver carp. These polymorphic loci should provide sufficient level of genetic diversity to evaluate population structure of bighead carp.     

Construction of a genetic linkage map for silver carp (Hypophthalmichthys molitrix)

For silver carp (Hypophthalmichthys molitrix), a combined microsatellite (or simple sequence repeat) and amplified fragment length polymorphism (AFLP) sex average linkage map was constructed. A total of 483 markers (245 microsatellites and 238 AFLPs) were assigned to 33 linkage groups. The map spanned 1352.2 cM, covering 86.4% of the estimated genome size of silver carp. The maximum and average spaces between 420 loci were 21.5 cM and 3.2 cM, respectively. The length of linkage groups ranged from 3.6 cM to 98.5 cM with an average of 41.0 cM. The number of markers per group varied from 2 to 44 with an average of 14.6. The AFLP markers significantly improved the integrity of microsatellite-based linkage groups and increased the genome coverage and marker evenness. A genome-wide recombination suppression was observed in male. In an extreme case, six microsatellites co-segregated in male, but spanned a 45.1 cM region in female.     

The effect of light stimuli on dark-adapted visual sensitivity in invasive silver carp Hypophthalmichthys molitrix and bighead carp H. nobilis

Non-physical barriers, including the use of underwater strobe lights alone or paired with sound or bubbles, are being considered as a means to prevent the upstream migration of invasive silver carp Hypophthalmichthys molitrix and bighead carp H. nobilis. To optimize potential optical deterrents, it is necessary to understand the visual sensitivity of the fishes. Dark-adapted H. molitrix and H. nobilis were found to possess broad visual sensitivity between 470 to 620 nm with peak spectral sensitivity at 540 nm for H. molitrix and 560 nm in H. nobilis. To assess the effect of a strobe light on vision, dark-adapted H. molitrix, H. nobilis and common carp Cyprinus carpio, were exposed to three different 5 s trains (100, 200, or 500 ms on–off flashes) of white light and the recovery of visual sensitivity was determined by measuring the b-wave amplitude of the electroretinogram (ERG). For all species, the longest recoveries were observed in response to the 500 ms flash trains (H. molitrix mean ± SE = 702.0 ± 89.8 s; H. nobilis 648.0 ± 116.0 s; C. carpio 480 ± 180.0 s). The results suggest that strobe lights can temporarily depress visual sensitivity, which may render optical barriers less effective.     

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.     

Cross‐species amplification in silver carp and bighead carp with microsatellite primers of common carp

About a third of microsatellite primers designed for common carp (Cyprinus carpio) was successfully amplified in silver carp (Hypophthalmichthys molitrix) and bighead carp (Aristichthys nobilis). These markers, inherited in Mendelian mode, are of potential applications in cypinid genetics.     

First U.K. recording in the wild of the bighead carp Hypophthalmichthys nobilis

Five specimens of bighead carp Hypophthalmichthys nobilis were captured from a lake in eastern England in January 2006, their first confirmed recording in the wild in the U.K. They varied in fork lengths from 746 to 796 mm and mass from 10·0 to 13·5 kg. All the fish were age 9 years and were slow growing compared to populations elsewhere in their expanded range.     

Development of silver carp (Hypophthalmichthys molitrix) and bighead carp (Aristichthys nobilis) genetic maps using microsatellite and AFLP markers and a pseudo-testcross strategy

Silver carp (Hypophthalmichthys molitrix) and bighead carp (Aristichthys nobilis) are two of the four most important pond-cultured fish species inhabiting the major river basins of China. In the present study, genetic maps of silver carp and bighead carp were constructed using microsatellite and AFLP markers and a two-way pseudo-testcross strategy. To create the maps, 60 individuals were obtained from a cross of a single bighead carp (female) and a single silver carp (male). The silver carp map consisted of 271 markers (48 microsatellites and 223 AFLPs) that were assembled into 27 linkage groups, of which 22 contained at least four markers. The total length of the silver carp map was 952.2 cM, covering 82.8% of the estimated genome size. The bighead carp map consisted of 153 markers (27 microsatellites and 126 AFLPs) which were organized into 30 linkage groups, of which 19 contained at least four markers. The total length of the bighead carp map was 852.0 cM, covering 70.5% of the estimated genome size. Eighteen microsatellite markers were common to both maps. These maps will contribute to discovery of genes and genetic regions controlling traits in the two species of carp.     

Biochemical genetic comparison of bighead (Hypophthalmichthys molitrix), silver carp (Aristichthys nobilis), and their hybrids reared in Czechoslovakia

Incidental and/or uncontrolled hybridization between silver carp ( Hypophthalmichthys molitrix ) and bighead ( Aristichthys nobilis ) represents a serious problem in Czechoslovak aquaculture. This fact affects fitness traits very negatively in successive generations of hybrids. To solve this problem, 1076 individuals in a total of both H. molitrix. A. nobilis , and their hybrids in 12 groups from six rearing facilities were analysed. Twelve protein systems representing 21 presumptive loci were studied to analyse the electrophoretic patterns of their products using horizontal starch gel electrophoresis of blood and tissue extracts. Both species displayed identical electrophoretic patterns in MYO-I* , LDH-A *, LDH-B *, sMDH-1 *, sIDHP-3*, GPI-1*, and CK-I * loci. For a reliable differentiation of both species products of the following nine loci are applicable ALB *, PA *, TF, sMDH-2 *, SOD *, NDH *, MYO-IF , MYO-III *, and CK-2 *. In addition, some polymorphic variants in slDHP-1 *, sIDHP-2 *, LDH-C *, EST-II *, and GPI-2 * loci are of use as auxillary markers while the other variants are common to both species. A high level of gene introgression was evident through hybridization between both species. All groups declared previously as ' H. molifrix ' were actually confirmed biochemically to be H. molitrix . However, all groups declared as ' A. nobilis ' were proved to be a mixture of A. nobilis and its hybrids of different level with H. molitrix . This suggests it is impossible to distinguish between A. nobilis and hybrids using their external morphology only.     

Swimming performance of adult bighead carp Hypophthalmichthys nobilis (Richardson, 1845) and silver carp H. molitrix (Valenciennes, 1844)

Although the movement of invasive bighead carp (Hypophthalmichthys nobilis) and silver carp (H. molitrix) in the Upper Mississippi River system is dependent on their ability to swim through its numerous lock‐and‐dams, the swimming performance of adults of these species is at present unknown. Using a large (2,935‐L) mobile swim tunnel, the swimming performance of adult bighead and adult silver carp was quantified at water velocities that challenged them to exhibit either prolonged and/or burst swimming (76–244 cm/s) with fatigue times of less than 10 min. Simple log‐linear models best described the relative swim speed to fatigue relationships for both species. Under these conditions, the swimming performances of adult bighead and silver carp were similar to several species of adult fishes native to the Mississippi River system, but relatively low (<3 total body lengths per second, TL/s) compared to previously studied juveniles and sub‐adult bigheaded carps (3–15 TL/s). The decline in endurance with water velocity was three times greater in bighead carp (slope = ?2.98) than in silver carp (slope = ?1.01) and the predictive ability of the bighead model was appreciably better than the silver carp model. The differences in adult swimming performance between the two species were coincident with behavioral differences (e.g. breaching in silver carp but not in bighead carp). The swimming performance data of adult bighead and silver carp can now be used to evaluate whether their passage through manmade river structures including the gates of lock‐and‐dams in the Upper Mississippi River might be reduced.     

Comparative studies on the feeding selectivity of silver carp Hypophthalmichthys molitrix and bighead carp Aristichthys nobilis

Silver carp and bighead carp exhibited size-selection for food particles in aquarium experiments, but did not select their preferred species of plankton actively when they were distributed evenly in the water. They also possessed the capacity of selection for feeding area. The removal rates (% g⁻¹ fish weight) of silver carp for smaller plankton (phytoplankton) were higher than those of bighead carp. The removal rates by the latter for bigger plankton (zooplankton) were higher than those of silver carp, but for plankton about 70 μm dia. the rates by the two species were almost equal.     

Characterization of microsatellite loci in silver carp (Hypophthalmichthys molitrix), and cross‐amplification in other cyprinid species

Captive populations of silver carp (Hypophthalmichthys molitrix), a major aquaculture species in Asia, would undoubtedly benefit from genetic monitoring and improvement programs. We report the isolation and preliminary characterization of 16 microsatellite loci derived from both conventional and microsatellite‐enriched libraries. Inheritance studies confirmed the allelic nature of observed polymorphisms at all loci, while identifying null alleles at two loci. These loci, having varying degrees of polymorphism, should provide useful markers for applied genetic studies. A high degree of cross‐amplification among 10 other cyprinid species suggests that these loci may have more widespread utility.     

A comparison of complete mitochondrial genomes of silver carp Hypophthalmichthys molitrix and bighead carp Hypophthalmichthys nobilis: implications for their taxonomic relationship and phylogeny

Based upon morphological characters, Silver carp Hypophthalmichthys molitrix and bighead carp Hypophthalmichthys nobilis (or Aristichthys nobilis ) have been classified into either the same genus or two distinct genera. Consequently, the taxonomic relationship of the two species at the generic level remains equivocal. This issue is addressed by sequencing complete mitochondrial genomes of H. molitrix and H. nobilis , comparing their mitogenome organization, structure and sequence similarity, and conducting a comprehensive phylogenetic analysis of cyprinid species. As with other cyprinid fishes, the mitogenomes of the two species were structurally conserved, containing 37 genes including 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA (tRNAs) genes and a putative control region (D-loop). Sequence similarity between the two mitogenomes varied in different genes or regions, being highest in the tRNA genes (98·8%), lowest in the control region (89·4%) and intermediate in the protein-coding genes (94·2%). Analyses of the sequence comparison and phylogeny using concatenated protein sequences support the view that the two species belong to the genus Hypophthalmichthys . Further studies using nuclear markers and involving more closely related species, and the systematic combination of traditional biology and molecular biology are needed in order to confirm this conclusion.     

中华鲟养殖池背角无齿蚌和鲢鳙鱼生态修复效果比较

2009年5月31日-6月20日在广东省大沙河水库利用微型生态系统比较不同放养密度的褶纹冠蚌(Cristaria plicata)和背角无齿蚌(Anodonta woodiana)对水体氮、磷及浮游植物的影响,探讨两种蚌在控制南亚热带水库富营养化水体藻类水华上的可行性。实验结果表明,在褶纹冠蚌和背角无齿蚌处理组中,总氮、总磷和叶绿素a浓度显著增加,而铵氮的浓度显著下降;褶纹冠蚌和背角无齿蚌导致了浮游植物群落组成结构的改变和数量的增加,实验过程中绿藻所占的比例迅速上升。两种蚌之间没有显著的差异,只是在不同的作用强度下,时间上的响应不同。综合实验结果,褶纹冠蚌和背角无齿蚌难以有效地运用于我国华南地区水库的水质改善与富营养化控制。     

Suction volume and filtering efficiency of silver carp (Hypophthalmichthys molitrix Val.) and bighead carp (Aristichthys nobilis Rich.)

Experiments were conducted to measure the suction volume of silver carp and bighead carp of age 1 + with respiratory chamber, and to calculate the suction volume and the filtering efficiency with respect to changes in concentrations of food particles. Suction volume (B. ml/mouth) and filtering efficiency (E. %) were calculated using the following formula: C ₁=C₀(1-BE/v)ⁿ where C₀ and C₁ were the concentrations of specific food particles at the beginning and at the end of experiment, respectively, V was the volume (ml) of experimental water, and n was the total number of observation of suction made during the experimental period. The relationships between suction volume (ml/mouth) of age I+ silver carp (B_h) and bighead carp (B_a) and their standard lengths (L, cm) were: B _h=0.561L-8.94, B_a= 0.627L-7.48 while those of the fingerlings were: B _h= O.l70L-0.837, B_a= 0.157L-0.418. The suction volume of the fingerlings was mainly affected by fish size, the function of temperature between 15 and 25° C being negligible. However, temperature affected filtering rate (filtered volume per unit time) through its effect on filtering frequency. The filtering efficiency of the fishes for rotifers (Brachionus caliciflorus) was 100 per cent. The relationships between filtering efficiency and sizes of food particles smaller than or equal to that of a rotifer were: E _h=25.1 ln e.s.d. -13.6, E_a=22.2 In e.s.d. -33.1 where E_h and E_a were filtering efficiency of silver carp and bighead carp, respectively, and e.s.d. was the equivalent spherical diameter (μm) of food particles.     

Native bighead carp Hypophthalmichthys nobilis and silver carp Hypophthalmichthys molitrix populations in the Pearl River are threatened by Yangtze River introductions as revealed by mitochondrial DNA

Bighead carp Hypophthalmichthys nobilis and silver carp Hypophthalmichthys molitrix have been two economically important aquaculture species in China for centuries. In the past decades, bighead and silver carp have been introduced from the Yangtze River to many river systems in China, including the Pearl River, in annual, large-scale, stocking activities to enhance wild fisheries. Nonetheless, few studies have assessed the ecological or genetic impacts of such introductions on native conspecific fish populations. An mtDNA D-loop segment of 978 bp from 213 bighead carp samples from 9 populations and a 975 bp segment from 204 silver carp samples from 10 populations were obtained to evaluate genetic diversity and population integrity. Results from a haplotype network analysis revealed that most haplotypes of the Pearl River clustered with those of Yangtze River origin and only a small proportion were distinct, suggesting that both the native Pearl River bighead and silver carp populations are currently dominated by genetic material from the Yangtze River. The genetic diversity of Pearl River populations is high in both species because of this inter-population gene flow, but the diversity of native Pearl River populations is low. To preserve the native genetic diversity, stocking of non-native fingerlings should cease immediately and native Pearl River bighead and silver carp fish farms should be established. This research demonstrates the danger to native biodiversity across China because of the substantial, ongoing stock-enhancement activities without prior genetic assessment.     

The hearing abilities of the silver carp (Hypopthalmichthys molitrix) and bighead carp (Aristichthys nobilis)

Concern regarding the spread of silver carp (Hypopthalmichthys molitrix) and bighead carp (Aristichthys nobilis) through the Illinois River has prompted the development of a Bio-acoustic Fish Fence (BAFF) to act as an acoustic fish deterrent. The application of this technology has resulted in a need to understand the auditory physiology of the target species, in order to maximise the effect of the barrier in preventing the migration of the non-indigenous carp species into Lake Michigan, whilst minimising the effect on indigenous fish populations. Therefore, the hearing thresholds of 12 H. molitrix and 12 A. nobilis were defined using the Auditory Brainstem Response (ABR) technique, in a pressure-dominated sound field generated by submerged transducers of the type used in the construction of the BAFF system. The results clearly show that these fish are most sensitive to sounds in a frequency bandwidth of between 750 Hz and 1500 Hz, with higher thresholds below 300 Hz and above 2000 Hz.     

利用17个微卫星标记分析鳙鱼的遗传多样性

选用本实验室克隆的17个鳙鱼微卫星分子标记分析四川泸州和江西鄱阳湖的两个种群鳙鱼的遗传多样性及种质特性,计算和统计了杂合度、多态信息含量（PIC）、有效等位基因数、等位基因频率、遗传距离、遗传相似系数、Hardy-Weinberg平衡偏离指数等方面内容。结果表明：选择使用17个微卫星标记,其中有4个为单态标记,13个为多态标记。江西和四川鳙鱼群体每个微卫星位点的平均等位基因数分别为3.325及3.882,平均有效等位基因数分别为3.531及2.676,多态位点百分率分别为82.4及70.5, 17个微卫星标记共有等位基因71个,多态微卫星位点的PIC在0.114~0.960之间变动,平均为0.417 ,两群体位点平均观测杂合度为0.385和0.452,平均期望杂合度为0.360和0.422,两个群体间的遗传相似系数为0.897,群体间的遗传距离为0.109。     

Digestive enzyme pattern of two stomachless filter feeders, silver carp, Hypophthalmichthys molitrix Val., and bighead carp, Aristichthys nobilis Rich.

Activities of digestive enzymes (trypsin and amylase) of microplanktophagous silver carp, Hypophthalmichthys molitrix , and macroplanktophagous bighead carp, Aristichthys nobilis , were investigated in relation to pH in 10 different segments of the gut. In addition, the possible existence of 'classical' lysozyme in both species and cellulase in silver carp were scrutinized, but no activities were detected. Tryptic and amylolytic activities decreased sharply from fore-gut to hind-gut indicating an efficient reabsorption mechanism. Trypsin and amylase of both species had a pH optimum at 8.3 and 7.0 respectively. The relatively low pH of 6.4 in the fore-gut of silver carp did not support an efficient operation of these enzymes, which was compensated by a higher enzyme concentration as compared to bighead carp. The latter, however, showed a higher pH of 7.3 in the fore-gut and thereby maintained similar activities as silver carp. The distinction of herbivorous and omnivorous fish in terms of digestion mechanisms is discussed.     

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.     

A second‐generation genetic linkage map for bighead carp (Aristichthys nobilis) based on microsatellite markers

Bighead carp (Aristichthys nobilis) is an important aquaculture fish worldwide. Genetic linkage maps for the species were previously reported, but map resolution remained to be improved. In this study, a second‐generation genetic linkage map was constructed for bighead carp through a pseudo‐testcross strategy using interspecific hybrids between bighead carp and silver carp. Of the 754 microsatellites genotyped in two interspecific mapping families (with 77 progenies for each family), 659 markers were assigned to 24 linkage groups, which were equal to the chromosome numbers of the haploid genome. The consensus map spanned 1917.3 cM covering 92.8% of the estimated bighead carp genome with an average marker interval of 2.9 cM. The length of linkage groups ranged from 52.2 to 133.5 cM with an average of 79.9 cM. The number of markers per linkage group varied from 11 to 55 with an average of 27.5 per linkage group. Normality tests on interval distances of the map showed a non‐normal marker distribution; however, significant correlation was found between the length of linkage group and the number of markers below the 0.01 significance level (two‐tailed). The length of the female map was 1.12 times that of the male map, and the average recombination ratio of female to male was 1.10:1. Visual inspection showed that distorted markers gathered in some linkage groups and in certain regions of the male and female maps. This well‐defined genetic linkage map will provide a basic framework for further genome mapping of quantitative traits, comparative mapping and marker‐assisted breeding in bighead carp.