SNP panel development for genetic management of wild and domesticated white bass (Morone chrysops)

White bass (Morone chrysops), striped bass and their interspecific hybrid are important game fishes, whereas the hybrid striped bass is an important aquaculture species in the US. Numerous state, federal and private hatcheries, therefore, rear these species for stocking purposes as well as for food fish. Although striped bass populations (both wild and domesticated) have been extensively evaluated, relatively little effort has been directed toward the study and improvement of white bass. In this study, we developed SNP resources to examine the genetic relationships among a long‐term domesticated white bass line and five potential founder stocks for selective breeding collected from drainages in Arkansas, Texas and Alabama. Using genotyping‐by‐sequencing, we generated 13 872 genome‐wide SNP loci across the six populations. Stringent filtering of SNP‐calling parameters identified 426 informative SNP loci. Population genetic and structure analyses using these loci revealed only moderate genetic differentiation between populations (global F_st = 0.083) and indicated two major genetic clusters. A final 57‐SNP assay was successfully designed and validated using the MassARRAY system. The developed SNP panel assigned 96 additional genotyped individuals to their population of origin with 100% accuracy. The SNP resources developed in this study should facilitate ongoing efforts in selective breeding and conservation of white bass.     

Microsatellite DNA markers for parental assignment in hybrid striped bass (Morone saxatilis × Morone chrysops)

Development of nine polymorphic microsatellites from a genomic library of hybrid striped bass (female Morone chrysops × male Morone saxatilus) DNA is described. Breeding of hybrid striped bass for aquaculture is based largely on breeding wild fish. Molecular markers such as microsatellites will be useful tools for developing broodstock, estimating heritability for production traits, and selective breeding via marker‐assisted selection. The nine polymorphic microsatellites include six dinucleotide and three complex repeat motifs. The number of alleles detected among a sample of 10 individuals of each species was relatively low. All polymerase chain reaction primer pairs also amplified products in the sea bass Dicentrarchus labrax.     

Isolation and characterization of 149 novel microsatellite DNA markers for striped bass,Morone saxatilis,and cross‐species amplification in white bass,Morone chrysops,and their hybrid

To support detailed genetic analysis of striped bass (Morone saxatilis) and white bass (Morone chrysops), we isolated 153 microsatellite loci from repeat‐enriched striped bass DNA libraries. Of these, 147 markers amplified in striped bass (average 4.7 alleles per locus) and 133 in white bass (average 2.2 alleles per locus). One hundred twenty‐two markers amplified in their hybrid. Development of new microsatellite markers will facilitate evaluations of genetic structure in wild populations and will support pedigree analysis and linkage mapping for selective breeding.     

Evolution of an MHC class Ia gene fragment in four North American Morone species

A nucleotide sequence analysis of a fragment of a Morone MHC class Ia gene detected high levels of polymorphism in striped bass Morone saxatilis, white perch Morone americana and yellow bass Morone mississippiensis. Extremely low levels of MHC diversity, however, were detected in white bass Morone chrysops, suggesting the possibility of a severe population bottleneck for this species.     

Identification and characterization of microsatellites for striped bass from repeat-enriched libraries

Striped bass (Morone saxatilis) is economically important in the US due to its value as an aquaculture species and in supporting commercial and recreational fisheries, especially those off the Atlantic coast and in the Gulf of Mexico. Modern strategies for managing fishery populations and aquaculture broodstocks employ the use of molecular genetic markers to identify individuals, assign parentage, and characterize population genetic structure and levels of inbreeding and migration. As part of a collaborative effort to utilize molecular genetic technologies in striped bass breeding programs we generated microsatellite markers for use in population genetic studies, broodstock selection and management strategies, and the construction of a genetic map. We developed 345 new microsatellite markers for striped bass, a subset (n=71) of which was characterized by genotyping samples from two striped bass broodstock populations to evaluate marker polymorphism, percent heterozygosity, Hardy–Weinberg equilibrium (HWE), linkage disequilibrium (LD) and utility for population genetic studies.     

A Microsatellite Linkage Map of Striped Bass (Morone saxatilis) Reveals Conserved Synteny with the Three-Spined Stickleback (Gasterosteus aculeatus)

The striped bass (Morone saxatilis) and its relatives (genus Morone) are of great importance to fisheries and aquaculture in North America. As part of a collaborative effort to employ molecular genetics technologies in striped bass breeding programs, we previously developed nearly 500 microsatellite markers. The objectives of this study were to construct a microsatellite linkage map of striped bass and to examine conserved synteny between striped bass and three-spined stickleback (Gasterosteus aculeatus). Of 480 microsatellite markers screened for polymorphism, 289 informative markers were identified and used to genotype two half-sib mapping families. Twenty-six linkage groups were assembled, and only two markers remain unlinked. The sex-averaged map spans 1,623.8 cM with an average marker density of 5.78 cM per marker. Among 287 striped bass microsatellite markers assigned to linkage groups, 169 (58.9%) showed homology to sequences on stickleback chromosomes or scaffolds. Comparison between the stickleback genome and the striped bass linkage map revealed conserved synteny between these two species. This is the first linkage map for any of the Morone species. This map will be useful for molecular mapping and marker-assisted selection of genes of interest in striped bass breeding programs. The conserved synteny between striped bass and stickleback will facilitate fine mapping of genome regions of interest and will serve as a new resource for comparative mapping with other Perciform fishes such as European sea bass (Dicentrarchus labrax), gilthead sea bream (Sparus aurata), and tilapia (Oreochromis ssp.).     

Use of cellular oncogene probes to identify Morone hybrids.

We tested the ability of cellular oncogene (c-onc) probes to identify F1 hybrids and the lineage of known backcrosses within the fish genus Morone. Total DNA was isolated from five to 14 individuals per North American Morone species (striped bass, white bass, white perch, and yellow bass). The DNA was digested with two restriction enzymes, Eco RI and Hin dIII, Southern blotted, and hybridized to six different c-onc probes including v-abl, v-erb B, c-myc, c-H-ras, c-K-ras, and v-src. We found fixed genotypic differences among the four species for all six probes in single restriction enzyme digests. The heritability of these nuclear DNA genotypes was evaluated in hatchery-produced F1 Morone hybrids (striped bass x white bass and striped bass x white perch) tested with the six informative single probe/restriction enzyme combinations. All F1 individuals exhibited heterozygosity in all diagnostic nuclear DNA fragments, confirming the Mendelian inheritance of these genotypes in these fish. Furthermore, analysis of these nuclear DNA genotypes in hatchery-produced backcrosses of F1 hybrids striped bass x (white bass x striped bass) detected both recombinant and parental genotypes at all six polymorphic c-onc sequences. The lineage of suspected Morone hybrids of unknown descent collected from Lewis Smith Lake, Alabama, and from the Occoquan River, Virginia, was determined using the c-onc probes. Our results suggest that c-onc probes are suitable markers to unequivocally identify F1 hybrids and backcrosses and to quantify introgression in natural populations of fishes. The addition of RFLP analysis of mtDNA provided a complete ancestral history of individual fish.     

Tissue essential fatty acid composition and competitive response to dietary manipulations in white bass (Morone chrysops), striped bass (M. saxatilis) and hybrid striped bass (M. chrysopsxM. saxatilis)

The effects of wide changes in dietary levels of docosahexaenoic (DHA) or arachidonic (ArA) acids on growth, survival and fatty acid composition in body tissues of Morone larvae were examined. White bass (WB, Morone chrysops), striped bass (SB, Morone saxatilis) and sunshine hybrid bass (HSB, M. chrysopsxM. saxatilis) larvae (day 24-46) were fed Artemia nauplii enriched with algal sources of varying proportions of DHA and ArA (from 0 to over 20% of total fatty acids). WB larvae fed DHA-deficient Artemia diet retarded over 50% of their potential growth, however, increasing dietary DHA/ArA ratios were associated with a significant growth improvement. The highest proportion of polyunsaturated fatty acids was found in WB neural tissue (approx. 50% of total fatty acids), while HSB neural tissue contained the highest proportion of saturated fatty acids (approx. 35% of total fatty acids). Within the neural tissues of all Morone larvae, both DHA and ArA were generally the most dominant as well as the most responding fatty acids to dietary manipulations (except in WB fed DHA or ArA deficient diets). HSB neural tissue was particularly efficient in retaining a significant amount of DHA in the face of dietary deficiency. However, WB neural tissue was the most responsive to dietary increase in DHA, accumulating a significantly higher amount of DHA (P<0.05) than SB or HSB. Results demonstrate significant differences in fatty acid composition and growth responsiveness to dietary manipulations between Morone larvae species and within specific tissues. WB weight gain and neural tissue composition was affected most by dietary changes in both DHA and ArA whereas SB and HSB tissue compositions were generally less affected by dietary manipulations.     

Isolation and Characterization of Microsatellite Loci for Striped Bass (Morone saxatilis)

Genetic variation has been difficult to detect in striped bass (Morone saxatilis). Therefore, we identified and characterized 13 microsatellite loci to provide additional genetic markers for striped bass. Microsatellites were identified by screening a striped bass genomic library or by using primers developed for European sea bass (Dicentrarchus labrax) microsatellite loci. We found that 6 of the 13 microsatellite loci were polymorphic in DNA samples obtained from wild populations of striped bass. The number of alleles per locus varied from 3 to 12, and the observed heterozygosities ranged from 0.55 to 0.78. These results indicate that microsatellite loci provide more alleles and higher heterozygosities than other genetic markers developed for striped bass. Received November 9, 1999; accepted February 11, 2000.     

Parasites and diseases of striped bass, Morone saxatilis (Walbaum), from the lower Chesapeake Bay

A survey to determine presence of parasitic and disease organisms and their effect on estuarine populations of striped bass, Morone saxatilis (Walbaum), was conducted monthly from May 1972 to May 1973. A total of 514 fish over 1 year old and 140 young-of-the-year were examined using standard necropsy and histological procedures. Other species of fishes were studied to determine the specificity of striped bass parasites and to determine if other fishes were reservoirs for striped bass pathogens. Forty-five species of parasitic organisms from viruses to Metazoa were recognized from striped bass. Heavy infections by some were associated with definite pathological conditions.     

Ontogeny of gonadotropin-releasing hormone (GnRH) neurons in hybrid striped bass Morone sp.: whole-mount in situ hybridization analysis

The ontogeny of gonadotropin-releasing hormone (GnRH) mRNA-producing neurons was studied in developing hybrid striped bass (white bass Morone chrysops female × striped bass Morone saxatilis male), 1–55 days post-fertilization (dpf), by whole-mount in situ hybridization. Neurons that produce salmon (s) GnRH were first detected at 32 h post-fertilization in the olfactory placodes. These neurons migrated posteriorly during development and reached their final position at the olfactory bulbs-telencephalon boundary, possibly the terminal nerve ganglion (TNg) by 11 dpf. First signal of chicken (c) GnRH-II neurons appeared in the midbrain 2 dpf and remained there throughout development. A signal of seabream (sb) GnRH mRNA was first detected at 21 dpf in the preoptic area (POA) and as a bilateral continuum along the ventral telencephalon at 32–55 dpf. The expression of all three forms of GnRH increased throughout development. These results suggest that cGnRH-II neurons originate in the mid-brain, and that sGnRH neurons originate in the olfactory placodes and migrate caudally to the TNg. Neurons that express sbGnRH seem to originate at the preoptic area and then to migrate anteriorly along the ventral telencephalon. An olfactory placodal origin of these neurons, however, cannot be ruled out.     

Comparative severity of experimentally induced mycobacteriosis in striped bass Morone saxatilis and hybrid tilapia Oreochromis spp

Twenty striped bass Morone saxatilis and 20 hybrid tilapia Oreochromis niloticus x O. mossambicus x O. aureus each received a single intramuscular injection of 1.6 x 10(6) colony forming units per gram body weight of Mycobacterium marinum. Striped bass manifested significantly greater clinical and microscopic disease compared to tilapia. Whereas all the striped bass had died or were clinically ill by Day 8 post-infection, there was no apparent disruption of normal behaviour, physical appearance, or growth in any of the sacrificed or surviving tilapia. Histologically, granulomas in striped bass were generally larger and less discrete, with a higher proportion of heavily vacuolated macrophages, and large cores of necrotic cells. Visceral granulomas in tilapia were smaller, with a higher proportion of epithelioid macrophages, more pigment-containing cells, more peripheral lymphocytes, and virtually no central necrosis. Visceral granulomas were 18-fold more numerous in striped bass than in tilapia. Based upon histomorphometric data, mean proportions of acid-fast bacteria within pronephros granulomas were 4-fold greater in striped bass than tilapia, and striped bass granulomas averaged more than twice as large as tilapia granulomas. In the anterior kidney of striped bass, a positive correlation existed between mean mycobacterial proportions and mean necrosis scores. In tilapia, mean mycobacterial proportions correlated negatively with mean granuloma numbers, whereas there was no correlation between these parameters in striped bass. Results suggest that intrinsic functional differences in the immunologic systems of striped bass and hybrid tilapia may contribute to inter-species variation in mycobacteriosis susceptibility.     

An in vitro assay to measure phagocytosis in striped bass hybrids (Morone saxatilis x Morone chrysops)

An in vitro phagocytosis assay was developed for hybrid striped bass (Morone saxatilis x Morone chrysops), using cells collected from the peritoneal cavity of this fish. The findings indicated that: (1) 10 days following a single intraperitoneal injection (1 ml) of Freund's incomplete adjuvant (FIA) was an appropriate time for collecting suitable working concentrations (5.3+/-4.0 x 10(7) cells ml(-1)) of peritoneal phagocytes (83.7+/-1.5% macrophages) from these hybrids held at 23 degrees C; (2) these cells phagocytosed latex beads (polystyrene microspheres 3.12 microm in diameter) after 30 min of in vitro incubation at room temperature (25+/-1 degrees C). The phagocytic ability and phagocytic capacity in a washed adherent layer exposure system were 67.2+/-2.76% and 4.14+/-0.35 beads phagocyte(-1), respectively. These results strongly suggest that a simple methodology, including baseline data serving as guidelines, is now available for conducting in vitro phagocytosis assays in this hybrid.     

Age‐0 striped bass,Morone saxatilis (Walbaum, 1792), response to immunostimulation

Young‐of‐the‐year (age‐0) striped bass, Morone saxatilis, were studied to characterize their responses to inflammatory stimuli. There were two studies, with the hypotheses that (i) <5 g striped bass would respond to inflammatory insult within 6 h, and (ii) response to antigens would be maintained for >24 h in larger fish. Study I was conducted to understand the cytokine expression pattern (IL‐1β, TNF‐α, Nramp and TGF‐β) in response to lipopolysaccharide (LPS) or Freund's complete adjuvant (FCA) stimulation in age‐0 striped bass (3.44 ± 1.68 g, 70.6 ± 10.3 mm fork length) up to 24 h post injection (hpi). Study II was similar to Study I, but striped bass were sampled over a longer time frame (by 48 hpi) and larger age‐0 striped bass were used (20.6 ± 5.9 g, 129.2 ± 10.9 mm fork length). It was confirmed that immunostimulants such as LPS and FCA induce production of inflammatory cytokines and Nramp, which are important in innate immune response to bacterial infection. The responses were rapidly stimulated with LPS (IL‐1β, TNF‐α, TGF‐β >3‐fold increase, compared to PBS) or FCA (IL‐1β >3‐fold and TGF‐β >2‐fold, compared to PBS) within 6 hpi and maintained in most cases 48 hpi (spleen Nramp and TGF‐β 2‐fold >PBS, at 24 and 48 h), similar to other teleosts. Intraperitoneal injection with PBS also simulated inflammatory gene expression, but was delayed (IL‐1β, TNF‐α, TGF‐β and Nramp, FCA and LPS< at 6 h; 24 h >LPS and PBS) in comparison to LPS and FCA, suggesting that this procedure and possibly the volume used can be stimulatory and potentially harmful in age‐0 fish. Therefore, this study suggests that age‐0 striped bass are capable of strong cytokine induction in response to immunological stimulation within a very short period of time.     

Apolipoprotein A-I from striped bass (Morone saxatilis) demonstrates antibacterial activity in vitro

HDL and apolipoprotein A-I from teleostean fishes demonstrate in vitro activity against gram-positive and gram-negative bacteria. In this study, we purified ApoA-1 from striped bass (Morone saxatilis) plasma and examined its in vitro antibacterial activity against Streptococcus sp., Escherichia coli, and Mycobacterium marinum. In addition, we obtained sequence for a putative striped bass ApoA-1 gene, which when translated contained the identical sequence generated from N-terminal sequencing of the purified ApoA-1. The predicted secondary and tertiary structures contained the characteristic proline residues and high alpha-helical content conserved between mammals and fishes. Purified ApoA-1 exhibited antibacterial activity against the bacteria assayed. Concentrations of 125 microg/mL for E. coli, 250 microg/mL for Streptococcus sp., and 250 microg/mL for M. marinum, inhibited bacterial growth by 50% compared to control. ApoA-1 plasma concentrations in experimental and wild fish ranged from undetectable levels to greater than 5 mg/mL, indicating that striped bass ApoA-1 is an effective antibacterial agent at concentrations below the range of physiological concentrations in striped bass plasma. We therefore conclude that ApoA-1 could play a role in innate defense against bacterial pathogens in striped bass.     

Contrasting population characteristics of yellow bass (Morone mississippiensis) in two southern Illinois reservoirs

Two southern Illinois reservoirs were investigated with contrasting size structures of yellow bass (Morone mississippiensis) to compare growth, mortality and recruitment patterns. Yellow bass were collected from Crab Orchard and Little Grassy lakes during April–May 2009 using AC electrofishing. Total length and weight were recorded and sagittal otoliths sectioned and aged by two readers. Increments between otolith annuli were measured and the Weisberg linear growth model was used to assess age and environmental (growth year) effects on individual growth for fish from the two lakes. Von Bertalanffy growth models indicated faster growth and a greater maximum total length for yellow bass in Little Grassy Lake. However, growth of fish in Little Grassy Lake nearly ceased after age 4. The Weisberg model indicated differences in individual growth rate between the two lakes that were consistent across years (age effects were significant, but growth year effects and the age‐growth year interaction were not). Inter‐lake differences in fish growth were present up to age 3. Recruitment was relatively stable in Crab Orchard, with year classes up to age 7 observed. Recruitment was more erratic in Little Grassy, with age 5 being the dominant year class and fish up to age 11 present. Differences in growth and recruitment patterns for yellow bass in the two lakes may be attributed to substantial inter‐lake differences in turbidity, morphoedaphic index, or yellow bass density. Maximum age of yellow bass (age 11) was higher than previously reported for this species, likely due to the use of otoliths to age fish rather than scales. This study provides baseline information on age and growth, mortality, recruitment, and size structure of yellow bass that can be compared to data in future studies to elucidate factors influencing population dynamics of this species.     

Conserved and Variant Molecular and Functional Features of Multiple Egg Yolk Precursor Proteins (Vitellogenins) in White Perch (Morone americana) and other Teleosts

Three complete cDNAs encoding different forms of vitellogenin (Vtg) were isolated from a white perch (Morone americana) liver cDNA library and characterized with respect to immunobiochemical and functional features of the three Vtgs and their product yolk proteins (YPs) in this species and in the congeneric striped bass (Morone saxatilis). The two longest cDNAs encoded Vtgs with a complete suite of yolk protein domains that, based on comparisons with vtg sequences from other species, were categorized as VtgAa and VtgAb using the current nomenclature for multiple teleost Vtgs. The shorter cDNA encoded a Vtg that lacked a phosvitin domain, had a shortened C-terminus, and was categorized as VtgC. Mapping of peptide sequences from the purified Vtgs and their derived YPs to Vtg sequences deduced from the cDNAs definitively identified the white perch VtgAa, VtgAb, and VtgC proteins. Detailed comparisons of the primary structures of each Vtg with partial or complete sequences of Morone yolk proteins or of Vtgs from other fishes revealed conserved and variant structural elements of teleost Vtgs with functional significance, including, as examples, signal peptide cleavage sites, dimerization sites, cathepsin D protease recognition sites, and receptor-binding domains. These comparisons also yielded an interim revision of the classification scheme for multiple teleost Vtgs. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Lesion induction by the plerocercoid Lacistorhynchus tenuis (Cestoda) and wound healing in the striped bass Morone saxatilis (Walbaum)

San Francisco Bay-Delta striped bass, Morone saxatilis (Walbaum), form open lesions in response to a plerocercoid infection of Lacistorhynchus tenuis (Van Beneden, 1858) (Cestoda: Trypanorhyncha). Laboratory infection experiments showed that striped bass can be infected with the plerocercoids by ingesting infected copepods. Histological sections indicated that a cellular host response was mounted early in the infection period, and that despite the leucocytic infiltration the parasites continued to develop. However, at 3 months post-infection some of the plerocercoids began to degenerate, and lesions formed at this time and 14 months post-infection. Open lesions in adult striped bass collected from the field took 2 months to heal and were detectable for at least 22 months. Regeneration of the muscle tissue did not occur although the wound completely healed externally.