Ecosystem responses to aquatic invasive species management: A synthesis of two decades of bigheaded carp suppression in a large river.

The invasion of silver carp (Hypophthalmichthys molitrix) and bighead carp (H. nobilis) or "bigheaded carps" has caused extensive ecological and economic harm throughout the Mississippi River and its tributaries. To prevent their continued spread upstream toward the Great Lakes, intense commercial harvest was implemented on the Illinois River, a large tributary that connects the Mississippi River to Lake Michigan. Since implementation, harvest has reduced densities at the invasion front while also presenting an opportunity to generate a synthesis on ecosystem resilience in the face of accelerating invasion. Resilience, the ability of an ecosystem to recover after perturbation, was observed at local scales and within some taxa but has yet to manifest at a river-wide scale and often co-varied with abiotic environmental or seasonal factors. Thus, while intensive harvest has limited further spread of bigheaded carps, and evidence of additional secondary ecosystem benefits exists, opportunities remain to identify potential pathways that could spread such ecosystem benefits even farther.

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.

Reduction of large vessel traffic improves water quality and alters fish habitat-use throughout a large river.

Rivers are increasingly used as superhighways for the continental-scale transportation of freight goods, but the ecological impact of large vessel traffic on river ecosystems is difficult to study. Recently, the temporary maintenance closure of lock and dam systems on the Illinois Waterway (USA) brought commercial vessel traffic to a halt along the river's length, offering a rare opportunity to study the response of the ecosystem before, during, and after an extended pause of this persistent anthropogenic disturbance. We observed improvements in main- and side-channel water quality and a redistribution of fish habitat-use during a months-long, near-complete reduction of large vessel traffic. Over 3600 water quality and 1300 fish community samples indicate that large vessel traffic reduction coincided with a 33 % reduction in turbidity as well as increased use of sampling strata near vessel navigation corridors by sound-sensitive and rheophilic fishes. Gizzard shad (Dorosoma cepedianum), the most abundant species in the system, also expanded their use of these 'impact' areas. Though inland waterway transport is an economically- and climate-friendly alternative to trucking and rail for the shipment of freight, our data suggest that intense vessel traffic may have profound physical and biological impacts across a large river. Monitoring and mitigation of ecological impacts of the ongoing expansion of inland waterway transport around the world will be critical to balancing large rivers as both useful navigation corridors and functional ecosystems.

New records of Spotted Bass, Micropterus punctulatus, within the Mississippi River Basin, Illinois.

Spotted Bass Micropterus punctulatus, like many sport fishes, have experienced range expansion through intentional introductions (i.e., legal stocking and illegal transfers) and migration across the United States. In Illinois, USA, native populations of Spotted Bass occur along the eastern and southern border of the state. We report new records of Spotted Bass in their non-native range of the Illinois Waterway and the Illinois portion of the Upper Mississippi River in addition to collections in their native range in the Illinois sections of the Ohio and Wabash rivers to better understand their current distribution. Continuous, collaborative efforts to track the distribution and expansion of non-native fishes are important for maintaining and establishing native and non-native fisheries management objectives and education, as non-native fishes can influence native species population distribution and dynamics.

Predation of invasive silver carp by native largemouth bass is size-selective in the Illinois River.

Silver carp (Hypophthalmichthys molitrix) are a nonnative, planktivorous, and highly invasive species of cyprinid located throughout the Mississippi River Basin. Although they co-occur with largemouth bass (Micropterus nigricans), an abundant native predatory fish, their predator-prey relationship is poorly understood. This potential relationship warrants investigation as largemouth bass are large-gaped predators capable of exhibiting top-down control on planktivorous fishes. The objectives of this study were to determine if largemouth bass consume juvenile silver carp, and if there was a relationship between length of largemouth bass and length of silver carp consumed. Largemouth bass were collected from the La Grange Pool of the Illinois River using 60 Hz-pulsed DC electrofishing and their diets were analyzed (n = 389, total length = 70-578 mm). Evidence of silver carp was present in 18% of diets of largemouth bass that consumed fish. Lengths of consumed silver carp were estimated from the dimensions of their recovered chewing pads or pharyngeal teeth in the stomachs of largemouth bass. A significant relationship between length of largemouth bass and length of silver carp consumed (p < 0.001, F = 34.63, r2 = 0.61) was observed. Estimated total lengths of silver carp were 34-101 mm and were recovered from diets of largemouth bass that were 94-262 mm total length. These results indicate enhancement of native largemouth bass populations is unlikely to substantially reduce silver carp populations in the Illinois River or in other waterways where these species co-occur.

Hydrological and lock operation conditions associated with paddlefish and bigheaded carp dam passage on a large and small scale in the Upper Mississippi River (Pools 14-18).

Movement and dispersal of migratory fish species is an important life-history characteristics that can be impeded by navigation dams. Although habitat fragmentation may be detrimental to native fish species, it might act as an effective and economical barrier for controlling the spread of invasive species in riverine systems. Various technologies have been proposed as potential fish deterrents at locks and dams to reduce bigheaded carp (i.e., silver carp and bighead carp (Hypophthalmichthys spp.)) range expansion in the Upper Mississippi River (UMR). Lock and Dam (LD) 15 is infrequently at open-river condition (spillway gates completely open; hydraulic head across the dam <0.4 m) and has been identified as a potential location for fish deterrent implementation. We used acoustic telemetry to evaluate paddlefish passage at UMR dams and to evaluate seasonal and diel movement of paddlefish and bigheaded carp relative to environmental conditions and lock operations at LD 15. We observed successful paddlefish passage at all dams, with the highest number of passages occurring at LDs 17 and 16. Paddlefish residency events in the downstream lock approach of LD 15 occurred more frequently and for longer durations than residency events of bigheaded carp. We documented upstream passages completed by two individual paddlefish through the lock chamber at LD 15, and a single bighead carp completed upstream passage through the lock chamber during two separate years of this study. We identified four bigheaded carp and 19 paddlefish that made upstream passages through the spillway gates at LD 15 during this study. The majority of the upstream passages through the spillway gates for both species occurred during open river conditions. When hydraulic head was approximately 1-m or greater, we observed these taxa opt for upstream passage through the lock chamber more often than the dam gates. In years with infrequent open-river condition, a deterrent placed in the downstream lock approach may assist in meeting the management goal of reducing upstream passage of bigheaded carps but could also potentially affect paddlefish residency and passage. Continued study to understand the effects of deterrents on native fish could be beneficial for implementing an integrated bigheaded carp control strategy. Understanding fish behavior at UMR dams is a critical information need for river managers as they evaluate potential tools or technologies to control upstream expansion of bigheaded carp in the UMR.

Gut microbiomes of bigheaded carps and hybrids provide insights into invasion: A hologenome perspective.

Gut microbiomes play an essential role in host survival and local adaptation and thus can facilitate the invasion of host species. Biological invasions have been shown to be linked to the genetic properties of alien host species. It is thus plausible that the holobiont, the host, and its associated microbiome act as an entity to drive invasion success. The bighead carp and silver carp (bigheaded carps), invasive species that exhibit extensive hybridization in the Mississippi River Basin (MRB), provided a unique model to test the holobiont hypothesis of invasion. Here, we investigated the microbiomes of foreguts and hindguts in bigheaded carps and their reciprocal hybrids reared in aquaculture ponds using 16S amplicons and the associated gene prediction. We found an admixed pattern in the gut microbiome community in bigheaded carp hybrids. The hybrid gut microbiomes showed special characteristics such as relatively high alpha diversity in the foregut, an increasing dissimilarity between foreguts and hindguts, and a remarkable proportion of genes coding for putative enzymes related to their digestion of main food resources (Cyanobacteria, cellulose, and chitin). The pond-reared hybrids had advantageous features in genes coding for putative enzymes related to their diet. The above results collectively suggested that the gut microbiomes of hybrids could be beneficial to their local adaptation (e.g., food resource utilization), which might have facilitated their invasion in the MRB. The gut microbial findings, along with the intrinsic genomic features likely associated with life-history traits revealed in our recent study, provide preliminary evidence supporting the holobiont hypothesis of invasion.

Resolving the genetic paradox of invasions: Preadapted genomes and postintroduction hybridization of bigheaded carps in the Mississippi River Basin.

The genetic paradox of biological invasions is complex and multifaceted. In particular, the relative role of disparate propagule sources and genetic adaptation through postintroduction hybridization has remained largely unexplored. To add resolution to this paradox, we investigate the genetic architecture responsible for the invasion of two invasive Asian carp species, bighead carp (Hypophthalmichthys nobilis) and silver carp (H. molitrix) (bigheaded carps) that experience extensive hybridization in the Mississippi River Basin (MRB). We sequenced the genomes of bighead and silver carps (~1.08G bp and ~1.15G bp, respectively) and their hybrids collected from the MRB. We found moderate-to-high heterozygosity in bighead (0.0021) and silver (0.0036) carps, detected significantly higher dN/dS ratios of single-copy orthologous genes in bigheaded carps versus 10 other species of fish, and identified genes in both species potentially associated with environmental adaptation and other invasion-related traits. Additionally, we observed a high genomic similarity (96.3% in all syntenic blocks) between bighead and silver carps and over 90% embryonic viability in their experimentally induced hybrids. Our results suggest intrinsic genomic features of bigheaded carps, likely associated with life history traits that presumably evolved within their native ranges, might have facilitated their initial establishment of invasion, whereas ex-situ interspecific hybridization between the carps might have promoted their range expansion. This study reveals an alternative mechanism that could resolve one of the genetic paradoxes in biological invasions and provides invaluable genomic resources for applied research involving bigheaded carps.

Body condition (Wr ) and reproductive potential of bighead and silver carp hybrids: Postzygotic selection in the Mississippi River Basin.

Invasive bighead (Hypophthalmichthys nobilis) and silver carp (H. molitrix) are reproductively isolated in their native range, but form a bimodal, multigenerational hybrid swarm within the Mississippi River Basin (MRB). Despite observed F1 hybrid superiority in experimental settings, effects of postzygotic selection on bighead and silver carp hybrids have not been tested in a natural system. Individual parent and hybrid genotypes were resolved at 57 species-specific loci and used to evaluate postzygotic selection for body condition (Wr ) and female reproductive potential (presence of spawning stage gonads and gonadosomatic index [GSI]) in the MRB during 2009-2011. Body condition in the Marseilles Reach, Illinois River declined with a decrease in species-specific allele frequency from 1.0 to 0.4 for each species and early generation hybrids (F1, F2, and first-generation backcross) had lower mean Wr than late generation hybrids (2nd+ generation backcrosses) and parentals. Proportions of stage IV and stage V (spawning stage) female gonads differed between bighead and silver carp, but not among parentals and their early and late generation hybrids within the MRB. Mean GSI values did not differ between parentals and hybrids. Because reproductive potential did not differ between hybrids and parentals, our results suggest that early generation hybrids occur in low frequency either as a factor of poor condition (Wr ) and postreproductive survival, infrequent reproductive encounters by parental bighead and silver carp, or selection pressures acting on juvenile or immature life stages. Our results suggest that a combination of genetic and environmental factors may contribute to the postzygotic success of bighead and silver carp hybrids in the Mississippi River Basin.

Differential gene expression in chronic wasting disease-positive white-tailed deer (Odocoileus virginianus).

Chronic wasting disease (CWD) is a transmissible spongiform encephalopathy (TSE) that affects cervid species throughout North America. We evaluated gene expression in white-tailed deer collected by Illinois Department of Natural Resource wildlife managers during annual population reduction (e.g., sharpshooting) and disease monitoring efforts throughout the CWD-endemic area of northcentral Illinois. We conducted comparative transcriptomic analysis of liver and retropharyngeal lymph node tissue samples between CWD-positive (n = 5) and CWD-not detected (n = 5) deer. A total of 74,479 transcripts were assembled, and 51,661 (69.36%) transcripts were found to have matched proteins in NCBI-NR and UniProt. Our analysis of functional categories showed 40,308 transcripts were assigned to at least one Gene Ontology term and 37,853 transcripts were involved in at least one pathway. We identified a total of 59 differentially expressed genes (DEGs) in CWD-positive deer, of which 36 and 23 were associated with liver and retropharyngeal lymph node tissues, respectively. Functions of DEGs lend support to previous relationships between misfolded PrP and cellular membranes (e.g., STXBP5), and internal cellular components. We identified several genes that suggest a link between CWD and retroviruses and identified the gene ADIPOQ that acts as a tumor necrosis factor (TNF) antagonist. This gene may lead to reduced production of TNF and impact disease progression and clinical symptoms associated with CWD (i.e., wasting syndrome). Use of candidate genes identified in this study suggests the activation of endogenous processes in CWD-positive deer, which in turn may enable earlier detection of the disease.

Transcriptomic comparison of invasive bigheaded carps (Hypophthalmichthys nobilis and Hypophthalmichthys molitrix) and their hybrids.

Bighead carp (Hypophthalmichthys nobilis) and silver carp (Hypophthalmichthys molitrix), collectively called bigheaded carps, are invasive species in the Mississippi River Basin (MRB). Interspecific hybridization between bigheaded carps has been considered rare within their native rivers in China; however, it is prevalent in the MRB. We conducted de novo transcriptome analysis of pure and hybrid bigheaded carps and obtained 40,759 to 51,706 transcripts for pure, F1 hybrid, and backcross bigheaded carps. The search against protein databases resulted in 20,336-28,133 annotated transcripts (over 50% of the transcriptome) with over 13,000 transcripts mapped to 23 Gene Ontology biological processes and 127 KEGG metabolic pathways. More transcripts were detected in silver carp than in bighead carp; however, comparable numbers of transcripts were annotated. Transcriptomic variation detected between two F1 hybrids may indicate a potential loss of fitness in hybrids. The neighbor-joining distance tree constructed using over 2,500 one-to-one orthologous sequences suggests transcriptomes could be used to infer the history of introgression and hybridization. Moreover, we detected 24,792 candidate SNPs that can be used to identify different species. The transcriptomes, orthologous sequences, and candidate SNPs obtained in this study should provide further knowledge of interspecific hybridization and introgression.

Restriction site-associated DNA sequencing generates high-quality single nucleotide polymorphisms for assessing hybridization between bighead and silver carp in the United States and China.

Bighead carp (Hypophthalmichthys nobilis) and silver carp (H. molitrix) are invasive species and listed as US federally injurious species under the Lacy Act. They have established populations in much of the Mississippi River Basin (MRB; Mississippi, Illinois, and Missouri rivers) and are capable of producing fertile hybrids and complex introgression. Characterizing the composition of this admixture requires a large set of high-quality, evolutionarily conserved, diagnostic genetic markers to aid in the identification and management of these species in the midst of morphological ambiguity. Restriction site-associated DNA (RAD) sequencing of 45 barcoded bighead and silver carp from the United States and China produced reads that were aligned to the silver carp transcriptome yielded 261 candidate single nucleotide polymorphisms (SNPs) with fixed allelic differences between the two species. We selected the highest quality 112 SNP loci for validation using 194 putative pure-species and F1 hybrids from the MRB and putative bighead carp and silver carp pure species from China (Amur, Pearl and Yangtze rivers). Fifty SNPs were omitted due to design/amplification failure or lack of diagnostic utility. A total of 57 species-diagnostic SNPs conserved between carp species in US and Chinese rivers were identified; 32 were annotated to functional gene loci. Twenty-seven of the 181 (15%) putative pure species were identified as hybrid backcrosses after validation, including three backcrosses from the Amur River, where hybridization has not been documented previously. The 57 SNPs identified through RAD sequencing provide a diagnostic tool to detect population admixture and to identify hybrid and pure-species Asian carps in the United States and China.

Substrate color-induced melanization in eight turtle species from four chelonian groups.

Background color convergence of prey occurring through local adaptation or phenotypically plastic responses can reduce predation rates by visual predators. We assessed the capacity for substrate color-induced melanization in eight turtle species within the groups Chelydridae, Emydidae, Kinosternidae, and Trionychidae by rearing individuals on black or white substrates for 160 days. In all aquatic turtle species, integuments of the head and carapace of the individuals that were reared on a black substrate were darker than of those reared on a white substrate. In the terrestrial turtle Terrapene carolina carolina, however, no significant differences in dorsal head skin or carapace color were observed between treatments. Histological examination of tail tips in three aquatic species (Chelydra serpentina serpentina, Graptemys geographica, and Trachemys scripta elegans) indicated that substrate color-induced melanization is morphological, involving the transfer of melanosomes from basal epidermal melanocytes to adjacent keratinocytes. Interestingly, substrate color-induced melanization in a previously studied Pleurodire species apparently involves physiological color change. We could not, however, rule out physiological components to color change in the turtles of our study.