Genotypes,haplotypes and diplotypes of <Emphasis Type="Italic">IGF</Emphasis>-<Emphasis Type="Italic">II</Emphasis> SNPs and their association with growth traits in largemouth bass (<Emphasis Type="Italic">Micropterus salmoides</Emphasis>)

Insulin-like growth factor II (IGF-II) is involved in the regulation of somatic growth and metabolism in many fishes. IGF-II is an important candidate gene for growth traits in fishes and its polymorphisms were associated with the growth traits. The aim of this study is to screen single nucleotide polymorphisms (SNPs) of the largemouth bass (Micropterus salmoides) IGF-II gene and to analyze potential association between IGF-II gene polymorphisms and growth traits in largemouth bass. Four SNPs (C127T, T1012G, C1836T and C1861T) were detected and verified by DNA sequencing in the largemouth bass IGF-II gene. These SNPs were found to organize into seven haplotypes, which formed 13 diplotypes (haplotype pairs). Association analysis showed that four individual SNPs were not significantly associated with growth traits. Significant associations were, however, noted between diplotypes and growth traits (P < 0.05). The fish with H1H3 (CTCC/CGCC) and H1H5 (CTCC/TTTT) had greater body weight than those with H1H1 (CTCC/CTCC), H1H2 (CTCC/TGTT) and H4H4 (TGCT/TGCT/) did. Our data suggest a significant association between genetic variations in the largemouth bass IGF-II gene and growth traits. IGF-II SNPs could be used as potential genetic markers in future breeding programs of largemouth bass.     

Variations in piscivory of invasive largemouth bass <Emphasis Type="Italic">Micropterus salmoides</Emphasis> associated with pond environments

Introductions of non-native predatory fishes can be a major driver of aquatic biodiversity loss. The largemouth bass Micropterus salmoides (L.) has been introduced throughout much of the world, thereafter negatively affecting native faunal communities owing to its predatory impact. To investigate the environmental factors affecting the predatory performance of invasive bass, we examined the stomach contents and habitat characteristics of bass in 15 irrigation farm ponds in northeastern Japan. The food habits of the bass populations differed among the studied ponds: the predominant prey items were fishes among bass in seven of the ponds, whereas aquatic invertebrates (mainly insects and zooplankton) were the predominant taxa in the diets of bass in the eight remaining ponds, with the onset of piscivory related to body size. The results of multivariate analysis indicated that the extent to which the bass consumed fish was positively associated with fish prey abundance and negatively associated with percentage of aquatic vegetation coverage. We suggest that the extent of aquatic vegetation coverage strongly influenced the predation efficiency of bass in the ponds. These findings might be employed to assess a pond ecosystem’s vulnerability to invasive largemouth bass and to reduce the predator’s impact on native fish species by improvements to the habitat.     

Association between expression levels and growth trait-related SNPs located in promoters of the <Emphasis Type="Italic">MC4R</Emphasis> and <Emphasis Type="Italic">MSTN</Emphasis> genes in <Emphasis Type="Italic">Spinibarbus hollandi</Emphasis>

Melanocortin 4 receptor: (MC4R) and Myostatin (MSTN) are two important growth trait-related genes in animals. In this study, we showed that two SNPs, MC4R-719A>G and MSTN-519C>T, found in the promoters of the MC4R and MSTN genes, respectively, are both associated with growth traits in Spinibarbus hollandi. Furthermore, we observed that there were significant associations between the expression levels of the MC4R and MSTN genes and these two growth trait-related SNPs. The expression level of MC4R gene in brain was lower in GG genotype fish with extremely high growth performance than that in AA genotype fish with extremely low growth performance. Expression level of the MSTN gene in muscle was lower in TT genotype fish with extremely high growth performance than that in CC and CT genotype fish with lower growth performance. The results indicated that these SNPs located in the promoters of MC4R and MSTN are associated with growth-related traits through modification of gene expression levels. The MSTN and MC4R SNPs may have useful application in effective marker-assisted selection aimed to increase output in S. hollandi.     

Fcγ receptor-mediated phagocytosis pathway was involved in phagocytosis of mIgM+ B lymphocytes from largemouth bass (Micropterus salmoides)

The potential for phagocytosis has been proven in teleost B cells, but the research on the regulatory mechanism of phagocytosis remains lacking. In this study, three largemouth bass (Micropterus salmoides) (15 ± 5 g) were injected intraperitoneally with Nocardia seriolae (10⁵ CFU/100 μl/fish) in vivo, and their spleen was collected at 72 h post-infection for mRNA-seq. After the de novo assembly of the paired-end reads, 73,622 unigenes were obtained. Gene expression profiling revealed that 2043 unigenes were differentially expressed after N. seriolae infection, comprising 1285 upregulated and 758 downregulated unigenes (q-value <0.05, log2FC > |2|) of which 181 genes were involved in phagocytosis. The Kyoto Encyclopaedia of Genes and Genomes (KEGG) analysis demonstrated that 12 differentially expressed genes (DEG) associated with phagocytosis were enriched in the Fcγ receptor-mediated phagocytosis signalling pathway. In vitro, the phagocytic ability of mIgM⁺ B lymphocytes was validated using indirect immunofluorescence assay (IIFA) and fluorescence activating cell sorter (FACS), and the phagocytosis rates of the mIgM⁺ B lymphocytes incubated with a Lyn inhibitor had decreased from 18.533 ± 6.00% to 11.610 ± 4.236% compared with the unblocked group. These results suggested that the Fcγ receptor-mediated phagocytosis signalling pathway had participated in the phagocytosis of B cells and provide further insight into the role of B cells in innate immunology.     

大口黑鲈转录组SNPs筛选及其与生长的关联分析

为开发人工饲料代替冰鲜杂鱼养殖大口黑鲈的分子标记,以食用冰鲜鱼和配合饲料的同批大口黑鲈为研究材料,利用RNA-Seq（RNA sequencing）技术挖掘SNPs（Single nucleotide polymorphisms）标记,并以关联分析筛选可用于育种的候选标记。转录组进行测序共获得174 M数据,8681个SNPs位点。挑选其中具有表达差异的50个SNPs位点进行SNaPshot分型,结果39个分型成功,其中有4个为假阳性,通过转录组技术开发出SNPs标记35个,成功率为70.0%。为进一步检验这些标记是否可用于评估驯食饲料的大口黑鲈选育研究,研究以327尾摄食人工配合饲料的大口黑鲈为试验材料,SPSS软件进行一般线性模型分析SNPs的不同基因型与生长性状的相关性,结果显示有2个SNPs位点与体质量、全长和体高等生长性状存在显著相关性（P<0.05）,可作为候选标记用于大口黑鲈的分子辅助育种。由于转录组数据直接反应基因的表达情况,从中挖掘与性状相关的优势基因型与分子标记的成功率高,效果较好。同时也为解决大口黑鲈选育研究中标记缺乏提供了有效途径,为选育提供遗传依据、加速育种进程。     

The energetic impact of overwinter prey assemblages on age-0 largemouth bass, <Emphasis Type="Italic">Micropterus salmoides</Emphasis>

Synopsis We compared survival, growth, and swimming performance of two size classes of age-0 largemouth bass, Micropterus salmoides, in the spring after being fed diets of bluegill, Lepomis macrochirus, fathead minnows, Pimephales promelas, or invertebrate prey during the winter. Regardless of prey assemblage, survival was uniformly high and independent of size. Length, wet- and dry-mass, and condition was also similar among treatments for both size classes. However, variation in individual performance differed, with the lowest variability in growth occurring among small age-0 largemouth bass in the invertebrate only treatment. Absolute and length corrected swimming speeds of largemouth bass were highest for invertebrate prey assemblages, intermediate for fathead minnow prey, and lowest for bluegill prey. The patterns in growth and spring swimming performance likely reflect the varied nutritive quality of different prey, the ability of largemouth bass to capture different prey, and competition with the piscine prey.     

Food habits and relative abundances of native piscivores: implications for controlling common carp

Common carp (Cyprinus carpio, carp) are a widespread and ecologically destructive invasive fish species. Carp management is critical for maintaining healthy aquatic ecosystems, and many control options are available, but most have proven to be ineffective. Carp abundances have increased at The Nature Conservancy’s Emiquon Preserve, Illinois, since its restoration in 2007 despite management efforts to suppress this species. We conducted a comparative diet study in Illinois, Tennessee, and Wisconsin to test whether bowfin (Amia calva), spotted gar (Lepisosteus oculatus, gar), and largemouth bass (Micropterus salmoides) commonly preyed upon carp. We focused on bowfin and gar because they are hypoxia-tolerant, similar to carp. We also assessed whether specific fish community characteristics were correlated with carp relative abundances. We found no evidence that bowfin, gar, and bass consumed large numbers of carp. However, carp may be limited in some ecosystems (e.g., Reelfoot Lake, Tennessee) through alternative mechanisms associated with bowfin, gar, bass, and bluegill (Lepomis macrochirus) included in a diverse native fish community.     

Growth,survival, and foraging abilities of early life history stages of blue tilapia,Oreochromis aureus,and largemouth bass,Micropterus salmoides

Synopsis I compared growth, survival, and foraging abilities of early life history stages of blue tilapia,Oreochromis (=Sarotherodon = Tilapia) aureus, and largemouth bass,Micropterus salmoides, at equivalent prey (zooplankton) densities in the laboratory. Survival and growth of blue tilapia exceeded those of largemouth bass and, at comparable ages, blue tilapia were superior to largemouth bass in ability to prey on zooplankton; the enhanced survival and growth of blue tilapia appeared to result from their larger initial (egg) size. Predatory abilities of the species at equivalent sizes were similar. The success of blue tilapia in colonizing new systems and rapidly achieving high abundances in Florida may be a consequence of large initial size.     

A genome-wide association study on growth traits in orange-spotted grouper (<Emphasis Type="Italic">Epinephelus coioides</Emphasis>) with RAD-seq genotyping

The orange-spotted grouper, Epinephelus coioides, is one of the most popular fish in China and Southeast Asian countries because of its important economic value. However, molecular mechanism underlying the growth of orange-spotted grouper has never been fully understood. Herein, we performed a genome-wide association study (GWAS) on a natural population of 198 individuals aiming to screen the whole genome of orange-spotted grouper for identification of growth-related loci by restrictionsite associated DNA sequencing. In this research, 261,366 single nucleotide polymorphisms (SNPs) were developed, in which 110 SNPs were identified to be correlated with growth and 20 SNPs were further confirmed to be associated with both body weight and total length. From these identified SNPs, we annotated a total of 34 genes, including adgrb2, csnkza1, cers5, col22a1, creb5, dnd1, dzank1, dnai1, npy2r, fat3, lrrk2, lrp5, map3k9, and so on. Among these candidate genes, npy2r (neuropeptide Y receptor Y2) was reported to play a critical role in growth of the orange-spotted grouper. In addition, population structure, principal component analysis, kinship matrix and linkage disequilibrium were examined to verify the accuracy and reliability of our GWAS results. Our data will also provide a valuable genetic resource for further marker-assisted selection program to improve growth quality in groupers.     

Phenotypic variation and vulnerability to predation in juvenile bluegill sunfish (<Emphasis Type="Italic">Lepomis macrochirus</Emphasis>)

Bluegill sunfish (Lepomis macrochirus) are known to diversify into two forms specialized for foraging on either limnetic or littoral prey. Because juvenile bluegills seek vegetative cover in the presence of largemouth bass (Micropterus salmoides) predators, natural selection should favor the littoral body design at size ranges most vulnerable to predation. Yet within bluegill populations, both limnetic and littoral forms occur where vegetation and predators are present. While adaptive for foraging in different environments, does habitat-linked phenotypic variation also influence predator evasiveness for juvenile bluegills? We evaluate this question by quantifying susceptibility to predation for two groups of morphologically distinct bluegills; a limnetic form characteristic of bluegills inhabiting open water areas (limnetic bluegill) and a littoral form characteristic of bluegills inhabiting dense vegetation (littoral bluegill). In a series of predation trials, we found that bluegill behaviors differed in open water habitat but not in simulated vegetation. In open water habitat, limnetic bluegills formed more dense shoaling aggregations, maintained a larger distance from the predator, and required longer amounts of time to capture than littoral bluegill. When provided with simulated vegetation, largemouth bass spent longer amounts of time pursuing littoral bluegill and captured significantly fewer littoral bluegills than limnetic fish. Hence, morphological and behavioral variation in bluegills was linked to differential susceptibility to predation in open water and vegetated environments. Combined with previous studies, these findings show that morphological and behavioral adaptations enhance both foraging performance and predator evasiveness in different lake habitats.     

Predation of Japanese dace, Tribolodon hakonensis, by largemouth bass, Micropterus salmoides, in experimental aquaria

 To test the size range of prey fish that largemouth bass, Micropterus salmoides, can successfully consume, live Japanese dace, Tribolodon hakonensis, were given as prey fish to individual largemouth bass in aquaria. The ratio of maximum standard length (SL) of the Japanese dace consumed by largemouth bass was 46–69% of bass SL. The maximum length of Japanese dace consumed did not differ significantly between largemouth bass and smallmouth bass (M. dolomieu) previously studied, although largemouth bass have relatively larger mouth sizes than smallmouth bass. Largemouth bass occasionally injured and killed Japanese dace larger than the limit that could be consumed.     

There must be something in the water: assessing the behavioral responses of rusty crayfish (<Emphasis Type="Italic">Orconectes rusticus</Emphasis>) to fish and amphibian predator kairomones

Animals use chemical cues to find food, locate mates, and detect potential predators. Detecting cues in a risky environment can induce behavioral changes to increase survival. Rusty crayfish (Orconectes rusticus) reduce activity, increase refuge use, and make defensive displays after detecting fish predator cues. However, no studies have introduced amphibian cues. We investigated crayfish responses to hellbender salamander (Cryptobranchus alleganiensis, a dominant predator of crayfish) cues and compared these to responses to largemouth bass (Micropterus salmoides) cues. Largemouth bass occur sympatrically with hellbenders and are known to induce distinct responses in rusty crayfish. We randomly assigned crayfish to predator cue and conspecific alarm cue combinations and recorded frozen behavior, appendage movement, locomotion, and refuge use. We found crayfish increased their proportion of time spent frozen and reduced their proportion of time spent active in the tank when exposed to either predator cue. Moreover, these responses were magnified when crayfish were exposed to predator cues in combination with conspecific alarm cues. Our experiment demonstrates evidence in support of the crayfish’s ability to detect and appropriately respond to predator cues alone and in combination with conspecific alarm cues. Future work should investigate the effects of these behavioral changes on trophic dynamics in a natural system.     

Distribution and diet of Suwannee bass and largemouth bass in the lower Santa Fe River,Florida

Synopsis Suwannee bass,Micropterus notius, and largemouth bass,Micropterus salmoides, were collected by electrofishing in six habitats in the lower Santa Fe River, Florida during May 1981–March 1982. Both bass species were collected concomitantly in all habitats and habitat segregation was not evident. Crayfish (Procambarus spp.) were the primary food of Suwannee bass. Fish were the primary food of largemouth bass, but crayfish were common in the diet of largemouth bass ≥300 mm long. Suwannee bass have a greater throat width and consumed longer and wider forage than did largemouth bass of equal length. Available evidence suggests that Suwannee bass exhibit a positive selection for crayfish and a diverse forage resource, including abundant crayfish, is necessary for a Suwannee bass to coexist with a largemouth bass. This is Journal Series Number 6034 of the Florida Agricultural Experiment Station.     

Fish behavior in elevated CO<Subscript>2</Subscript>: implications for a movement barrier in flowing water

Preventing the spread of invasive fishes is an important aspect of management programs, but is challenging due to the behavior of fish and the nature of aquatic environments. The use of dissolved carbon dioxide (CO₂) has recently gained traction as a non-physical barrier for invasive fishes due to its ability to elicit avoidance behaviors in fish. Research to date has focused on the development of CO₂ barriers using static water environments. Because CO₂ barriers have been proposed for flowing water (i.e., in rivers or shipping canals), understanding the dynamics between fish and elevated CO₂ in flowing water is essential. Our study aims to define threshold levels required to alter behavior of bluegill (Lepomis macrochirus) and largemouth bass (Micropterus salmoides) in flowing water, and to quantify behavioral metrics of fish exposed to < 200 [ambient], 25,000, 50,000, and 100,000 µatm pCO₂. We also sought to quantify the impacts of repeated CO₂ exposure on fish behavior. Bluegill showed increased activity at 25,000 µatm, while largemouth bass showed increased activity at 100,000 µatm. When repeatedly exposed to cycles of 50,000 µatm pCO₂, bluegill exhibited increased activity followed by a diminished response after the second exposure. Results from this study define threshold levels required to elicit behavioral responses, and show that the effects that multiple exposures of elevated pCO₂ can decline, possibly due to habituation. Results will help shape the development and deployment of a CO₂ barrier to control the movements of invasive fishes.