Diagnostic SNPs reveal widespread introgressive hybridization between introduced bighead and silver carp in the Mississippi River Basin

Hybridization among conspecifics in native and introduced habitats has important implications for biological invasions in new ecosystems. Bighead (Hypophthalmichthys nobilis) and silver carp (H. molitrix) are genetically isolated and occur in sympatry within their native range. Following their introduction to North America, however, introgressant hybrids have been reported throughout their expanded range within the Mississippi River Basin (MRB). The extent of introgression, both spatially and generationally, is largely unknown. Therefore, we examined mixed‐species populations from across the MRB to characterize the extent of interspecific gene flow. We assayed 2798 individuals from nine locations with a suite of species‐diagnostic SNPs (57 nuclear and one mitochondrial). Forty‐four per cent (n = 1244) of individuals displayed hybrid genotypes. Moreover, the composition of hybrid genotypes varied among locations and represented complex hybrid swarms with multiple generations of gene flow. Introgressive hybrids were identified from all locations, were bidirectional and followed a bimodal distribution consisting primarily of parental or parental‐like genotypes and phenotypes. All described hybrid categories were present among individuals from 1999 to 2008, with parents and later‐generation backcrosses representing the largest proportion of individuals among years. Our mitochondrial SNP (COII), tested on a subset of 730 individuals, revealed a silver carp maternal bias in 13 of 21 (62%) F₁ hybrids, in all silver carp backcrosses, and maintained throughout many of the bighead carp backcrosses. The application of this suite of diagnostic markers and the spatial coverage permits a deeper examination of the complexity in hybrid swarms between two invasive, introduced species.     

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.     

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.     

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.     

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.     

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.     

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.     

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.     

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.     

控(微囊)藻鲢、鳙排泄物光能与生长活性

利用叶绿素荧光技术,通过原位培养滤食性鱼类鲢(Hypophthalmichthys molitrix)、鳙(Aristichthys nobilis)摄食微囊藻(Microcysis)后的排泄物,研究鲢、鳙滤食对藻类光能及生长活性的影响。结果表明,微囊藻经鲢、鳙滤食后,排泄当天,其叶绿素荧光参数PSⅡ最大光能转化效率(Fv/Fm)、PSⅡ潜在光合活性(Fv/Fo)、PSⅡ实际光能转化效率(Yield)和光合电子传递速率(ETR)显著低于对照组(P0.05),而光化学猝灭(qP)和非光化学猝灭(NPQ)显著高于对照组(P0.05)。随后,微囊藻的光合活性逐渐恢复,鲢、鳙组的Fv/Fm、Fv/Fo、Yield和qP分别在第3天和第5天开始呈上升趋势,至实验结束时显著高于对照组,其中鲢组高于鳙组。而NPQ呈下降趋势,第13 d极显著低于对照组(P0.01)。原位培养期间,鲢、鳙组藻细胞密度和叶绿素a(Chl a)浓度呈增长趋势,且鲢组明显高于鳙组;鳙组藻类游离胞外多糖含量增长幅度高于鲢组。实验结束时,鲢、鳙组浮游藻类总生物量分别为对照组7.78、6.55倍,绿藻(Chlorophyta)和硅藻(Bacillariophyta)相对生物量增加,但微囊藻贡献率达93%以上。相关性分析显示,鲢组藻细胞密度、Chl a浓度和胞外多糖含量与Fv/Fm、Fv/Fo、Yield、ETR、qP均极显著正相关(P0.01),而与NPQ呈极显著负相关(P0.01)。鳙组藻细胞密度、Chl a浓度和胞外多糖含量仅与ETR呈极显著正相关(P0.01),与NPQ呈极显著负相关(P0.01)。可见,鲢、鳙单次滤食未对微囊藻造成生理上的致命损伤,藻类由于超补偿生长,其光合及生长活性在短期恢复并显著增强,有潜在加速水体富营养化可能,鲢、鳙增殖放流以控制蓝藻水华的措施值得商榷。     

Confocal microscopy as a useful approach to describe gill rakers of Asian species of carp and native filter‐feeding fishes of the upper Mississippi River system

To better understand potential diet overlap among exotic Asian species of carp and native species of filter‐feeding fishes of the upper Mississippi River system, microscopy was used to document morphological differences in the gill rakers. Analysing samples first with light microscopy and subsequently with confocal microscopy, the three‐dimensional structure of gill rakers in Hypophthalmichthys molitrix, Hypophthalmichthys nobilis and Dorosoma cepedianum was more thoroughly described and illustrated than previous work with traditional microscopy techniques. The three‐dimensional structure of gill rakers in Ictiobus cyprinellus was described and illustrated for the first time.     

50年来长江鲢、鳙形态特征的变迁(英文)

采用多元分析方法,对采自长江中游的上世纪中期(下称早期)和本世纪初(下称当前)的121尾1龄鲢、鳙的30项形态度量性状进行了分析。依据对判别贡献最大的5个参数所建立的判别公式,对鲢、鳙早期和当前群体的平均判别准确率分别达94.2%和98.%,显示鲢或鳙的当前与早期群体间在总体上存在着显著差异。单因素方差分析显示,在早期和当前群体间,鲢有12个特征差异极显著、8个差异显著,鳙分别有8个特征差异极显著或显著。其中,鲢和鳙的6个显著或极显著变大的特征参数集中在头部,而14个鲢和10个鳙的显著或极显著变小的特征参数则主要集中在躯干部与尾部。研究表明,最近50年来长江鲢、鳙的头及头部特征有相对变大的趋势,而躯干部与尾部的许多特征参数则有相对变小的趋势,这些形态演变趋势符合人们喜食鱼头、追求高经济效益的期望。     

珠江东塔产卵场鳙繁殖的生态水文需求

鳙(Aristichthys nobilis(Richardson)),我国著名的四大家鱼之一,是一种重要的大型淡水经济鱼类。然而近年来由于各种原因,我国鳙种群数量急剧下降。依据2006—2013年珠江中下游长时间序列仔鱼数据和日平均流量数据,分析了研究水域鳙的繁殖生态,包括仔鱼出现的时间分布特征和早期资源周年变化规律,同时采用交互小波光谱分析方法 (cross wavelet analysis),分析了径流量与鳙仔鱼多度的关系。发现珠江水系鳙早期资源发生主要在5至8月,2006—2011年早期补充资源量逐渐减少,2012—2013年间呈明显著增加;鳙早期资源年度出现的时间呈缩减趋势,其中最大繁殖量出现的时间逐年提前。交互小波光谱分析结果表明:径流量的改变与鳙仔鱼的多度显著正相关,当径流量大于5×103m3/s,且持续2d以上,是珠江水系鳙大量繁殖的基础,将对受梯级水坝控制的鳙产卵场的繁殖生态水文保障具有指导意义。     

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.     

Direct and indirect analysis of the fitness of Chrysochus (Coleoptera: Chrysomelidae) hybrids

The chrysomelid beetles Chrysochus auratus and C. cobaltinus form a narrow hybrid zone in western North America. We used a combination of direct and indirect analyses to examine the fitness of Chrysochus hybrids. For the direct analyses, we compared the mating frequency, longevity, fecundity and fertility of hybrid females and parentals. For the indirect approach, we tested predictions of multilocus genotype frequencies at a focal site in the hybrid zone, based on the frequencies of mating combinations during the previous generation. Hybrid females produced fewer eggs than did parentals and the eggs they produced in the lab failed to hatch, in contrast to those of parental females. In addition, contrary to predictions that 15.8% of the individuals at the focal site would have multilocus genotypes other than those expected of parentals or F₁ individuals, we found no such genotypes at this site. This hybrid zone appears to be an example of a classic tension zone, with endogenous selection against hybrid individuals. We discuss the implications of low hybrid fitness for the evolution of premating barriers in this system, and argue that the integration of direct and indirect approaches is a powerful means of assessing the relative fitness of hybrids, particularly for species in which mate choices are easy to observe in the field.  © 2005 The Linnean Society of London, Biological Journal of the Linnean Society , 2005, 84 , 273–286.     

Investigating temporal changes in hybridization and introgression in a predominantly bimodal hybridizing population of invasive sika (Cervus nippon) and native red deer (C. elaphus) on the Kintyre Peninsula,Scotland

We investigated temporal changes in hybridization and introgression between native red deer (Cervus elaphus) and invasive Japanese sika (Cervus nippon) on the Kintyre Peninsula, Scotland, over 15 years, through analysis of 1513 samples of deer at 20 microsatellite loci and a mtDNA marker. We found no evidence that either the proportion of recent hybrids, or the levels of introgression had changed over the study period. Nevertheless, in one population where the two species have been in contact since ～1970, 44% of individuals sampled during the study were hybrids. This suggests that hybridization between these species can proceed fairly rapidly. By analysing the number of alleles that have introgressed from polymorphic red deer into the genetically homogenous sika population, we reconstructed the haplotypes of red deer alleles introduced by backcrossing. Five separate hybridization events could account for all the recently hybridized sika‐like individuals found across a large section of the Peninsula. Although we demonstrate that low rates of F1 hybridization can lead to substantial introgression, the progress of hybridization and introgression appears to be unpredictable over the short timescales.     

Characterization of microsatellite loci in Spartina species (Poaceae)

The cordgrasses in the genus Spartina have become model organisms for studying biological invasions from both ecological and genetic perspectives. Here we characterize 11 disomic loci in Spartina alterniflora that show promise for population studies and for studying hybridization events between S. alterniflora and S. foliosa. Comparisons among invasive and native S. alterniflora populations showed that levels of allelic variation are lower in invasive populations. In addition, nearly all loci that amplified in S. foliosa populations and in a swarm of S. alterniflora×foliosa hybrids were polymorphic. We also found that several loci amplified successfully in other Spartina species.     

Modelling bioenergetic and population-level impacts of invasive bigheaded carps (Hypophthalmichthys spp.) on native paddlefish (Polyodon spathula) in backwaters of the lower Mississippi River

1. While invasions of large rivers by exotic fish species are well documented, assessing actual or potential impacts on native species is a challenge. Rapid assessments may be possible through the application of a combination of bioenergetic and population dynamic models.
2. Paddlefish (Polyodon spathula) is a native species in the central USA with a history of population decline due to waterway development and overharvesting for roe. It is not known whether paddlefish are impacted by resource competition from invasive bigheaded carp populations, including silver (Hypophthalmichthys molitrix) and bighead carp (Hypophthalmichthys nobilis), which have expanded dramatically in the Mississippi River.
3. We used bioenergetic models to project the potential impact of invasive silver and bighead carp on zooplankton density and paddlefish somatic growth in backwater habitat. Bioenergetic outputs were translated to impacts on fecundity, becoming inputs for 50-year metapopulation simulations of backwater habitat connected to the main-stem Mississippi River by episodic flood events.
4. Competition with carp reduced growth and increased the risk of population decline for paddlefish. Impacts increased disproportionately with increased carp abundance and were further exacerbated in scenarios with increased diet overlap or decreased zooplankton abundance.
5. We also analysed paddlefish condition data collected at sites near the lower Mississippi River with varying histories of carp invasion. These data give credence to the bioenergetic model output; paddlefish had reduced body condition at sites with long-established, high-density carp populations.
6. We conclude that invasive bigheaded carps have great potential to reduce paddlefish growth, fecundity, and abundance. The pairing of bioenergetics and population models is likely to be broadly useful in assessing the risks posed by other invasive species.