Mechanisms of Egg Yolk Formation and Implications on Early Life History of White Perch (Morone americana)

The three white perch (Morone americana) vitellogenins (VtgAa, VtgAb, VtgC) were quantified accurately and precisely in the liver, plasma, and ovary during pre-, early-, mid-, and post-vitellogenic oocyte growth using protein cleavage-isotope dilution mass spectrometry (PC-IDMS). Western blotting generally mirrored the PC-IDMS results. By PC-IDMS, VtgC was quantifiable in pre-vitellogenic ovary tissues and VtgAb was quantifiable in pre-vitellogenic liver tissues however, neither protein was detected by western blotting in these respective tissues at this time point. Immunohistochemistry indicated that VtgC was present within pre-vitellogenic oocytes and localized to lipid droplets within vitellogenic oocytes. Affinity purification coupled to tandem mass spectrometry using highly purified VtgC as a bait protein revealed a single specific interacting protein (Y-box binding protein 2a-like [Ybx2a-like]) that eluted with suramin buffer and confirmed that VtgC does not bind the ovary vitellogenin receptors (LR8 and Lrp13). Western blotting for LR8 and Lrp13 showed that both receptors were expressed during vitellogenesis with LR8 and Lrp13 expression highest in early- and mid-vitellogenesis, respectively. The VtgAa within the ovary peaked during post-vitellogenesis, while VtgAb peaked during early-vitellogenesis in both white perch and the closely related striped bass (M. saxatilis). The VtgC was steadily accumulated by oocytes beginning during pre-vitellogenesis and continued until post-vitellogenesis and its composition varies widely between striped bass and white perch. In striped bass, the VtgC accounted for 26% of the vitellogenin-derived egg yolk, however in the white perch it comprised only 4%. Striped bass larvae have an extended developmental window and these larvae have yolk stores that may enable them to survive in the absence of food for twice as long as white perch after hatch. Thus, the VtgC may play an integral role in providing nutrients to late stage fish larvae prior to the onset of exogenous feeding and its composition in the egg yolk may relate to different early life histories among this diverse group of animals.     

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.     

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.     

Maturation and fecundity of a stock-enhanced population of striped bass in the Savannah River Estuary, U.S.A

The striped bass Morone saxatilis population in the Savannah River (south-eastern U.S.A.) collapsed in the 1980s, and recent eorts to restore the population have resulted in increased catch-per-unit-eort (CPUE) of striped bass in the Savannah River Estuary (SRE). The abundance of eggs and larvae, however, remain well below historic levels. The primary cause of the population decline was remedied, and environmental conditions seem suitable for striped bass spawning. Regression analysis of data derived from ultrasonic imaging of 31 striped bass resulted in a statistical model that predicted ovary volume well (r²=0.95). The enumeration of oocytes from ovarian tissue samples and the prediction of ovary volume allowed fecundity to be estimated without sacrificing the fish. Oocyte maturation in Savannah River striped bass seemed to progress normally, with oocytes developing to final stages of maturity in larger fish (>750 mm L _T). Additionally, fecundity estimates were comparable to a neighbouring striped bass population. The environmental cues needed to trigger development and release of striped bass oocytes into the SRE appeared to be present. If most of the striped bass females in the SRE are still young (<7 years), the ability to produce large numbers of eggs will be limited. As these young fish mature, egg production probably will increase and the density of striped bass eggs eventually will approach historic levels, provided suitable habitat and water quality are maintained.     

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.     

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.).     

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.     

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.     

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.     

A phylogeny of the temperate seabasses (Moronidae) characterized by a translocation of the mt-nd6 gene

The entire mitochondrial genome of the striped bass Morone saxatilis was sequenced together with the mitochondrial (mt) control regions of the white bass Morone chrysops, white perch Morone americana, yellow bass Morone mississippiensis, spotted seabass Dicentrarchus punctatus, European seabass Dicentrarchus labrax and the Japanese seabass Lateolabrax japonicus. The resultant 17 580 base pair circular genome of M. saxatilis contains 38 genes (13 proteins, 23 transfer RNAs and two ribosomal RNAs) and a control region bordered by the proline and phenylalanine mitochondrial tRNAs. Gene arrangement was similar to other vertebrates, except that the mt-nd6 gene was found within the control region rather than the canonical position between the mt-nd5 and mt-cyb genes. This translocation was found in all the Morone and Dicentrarchus species studied without functional copies or pseudogenes in the ancestral position. In L. japonicus, the mt-nd6 gene was found in the canonical position without evidence of an mt-nd6 gene in the control region. A Bayesian analysis of these and published mt-nd6 sequences from 45 other Perciformes grouped the Morone and Dicentrarchus species monophyletically with a probability of 1·00 with respect to L. japonicus and all other perciforms, and placed the Dicentrarchus species in the basal position. These data reinforce current placement of L. japonicus outside the Moronidae and provide a clear evolutionary character to define this family. The phylogeny of the Moronidae presented here also supports the hypothesis of an anadromous common ancestor to this family that gave rise to the North American estuarine and freshwater species. A series of tandem repeats previously reported in M. saxatilis was found in the control region of all Morone species between the mt-nd6 and mt-rnr1 genes, but not in either Dicentrarchus species, which reinforces the continued use of these two separate genera.     

Comparative analysis of Hox paralog group 2 gene expression during Nile tilapia (Oreochromis niloticus) embryonic development

The hindbrain and pharyngeal arch-derived structures of vertebrates are determined, at least in part, by Hox paralog group 2 genes. In sarcopterygians, the Hoxa2 gene alone appears to specify structures derived from the second pharyngeal arch (PA2), while in zebrafish (Danio rerio), either of the two Hox PG2 genes, hoxa2b or hoxb2a, can specify PA2-derived structures. We previously reported three Hox PG2 genes in striped bass (Morone saxatilis), including hoxa2a, hoxa2b, and hoxb2a and observed that only HoxA cluster genes are expressed in PA2, indicative that they function alone or together to specify PA2. In this paper, we present the cloning and expression analysis of Nile tilapia (Oreochromis niloticus) Hox PG2 genes and show that all three genes are expressed in the hindbrain and in PA2. The expression of hoxb2a in PA2 was unexpected given the close phylogenetic relationship of Nile tilapia and striped bass, both of which are members of the order Perciformes. A reanalysis of striped bass hoxb2a expression demonstrated that it is expressed in PA2 with nearly the same temporal and spatial expression pattern as its Nile tilapia ortholog. Further, we determined that Nile tilapia and striped bass hoxa2a orthologs are expressed in PA2 well beyond the onset of chondrogenesis whereas neither hoxa2b nor hoxb2a expression persist until this stage, which, according to previous hypotheses, suggests that hoxa2a orthologs in these two species function alone as selector genes of PA2 identity.     

Spatial separation of fishes captured in passive gear in a turbid prairie lake

Synopsis Spatial separation of fishes in the littoral zone of a turbid prairie lake (Clear Lake, Iowa) was assessed with gill nets and fyke nets. Catch per unit of effort was used to determine differences among habitat types, sampling times within a 24 h period, and sampling months. Four of 10 species examined were significantly more numerous in one of the three habitats — nonvegetated, vegetated, or gravel-rock substrate. Black bullhead (Ictalurus melas) and bigmouth buffalo (Ictiobus cyprinellus) were most abundant in vegetated areas, yellow bass (Morone mississippiensis) in gravel-rock areas, and channel catfish (Ictalurus punctatus) in both non-vegetated and gravel-rock areas. Temporal patterns in habitat use were indicated for these four species, as well as yellow perch (Perca flavescens), white bass (Morone chrysops), common carp (Cyprinus carpio), walleye (Stizostedion vitreum vitreum), black crappie (Pomoxis nigromaculatus), white sucker (Catostomus commersoni). Journal Paper No. J-11039 of the Iowa Agriculture and Home Economics Experiment Station, Ames, Iowa. Project No. 2345. Financed by the U.S. Department of the Interior Office of Water Research and Technology and Iowa State University.     

Dietary shifts in a stressed fish assemblage: Consequences of a bivalve invasion in the San Francisco Estuary

We compared dietary patterns within a temperate estuarine fish assemblage (Suisun Marsh, CA, U.S.A.) during a period of high mysid shrimp abundance and after a major decline in mysid abundance caused by the invasion of the overbite clam Potamocorbula amurensis. Prior to the invasion, high dietary overlap, high stomach fullness, and low niche breadth occurred among the fishes in spring when mysid populations were high. Dietary overlaps decreased and niche breadth increased for all species but the endemic splittail Pogonichthys macrolepidotus in fall when mysid populations were low. Eight native species exhibited lower overall collective overlaps and fuller stomachs than five alien species, suggesting more efficient resource partitioning. After mysid abundance declined, only alien striped bass Morone saxatilis preyed upon mysids in greater than trace amounts. An alien mysid became an important prey for small striped bass, but striped bass also switched to piscivory at a smaller size than when mysids were abundant. Eight of 13 species exhibited significant declines in abundance during the study period, which were concordant with the original importance of mysids in their diets. Our results suggest that altered lower food web dynamics in the San Francisco Estuary caused by the invasion of the overbite clam changed fish diets and have contributed to declines in fish abundance.     

Interactions between bluefish and striped bass: Behavior of bluefish under size- and number-impaired conditions and overlap in resource use

A decline in bluefish (Pomatomus saltatrix L.) recreational landings during the 1990s and the early 2000s led to multiple theories on the ultimate cause. One theory was that a large portion of the bluefish population moved offshore and was unavailable to nearshore recreational fishers; one reason given for the movement offshore was increased competition with striped bass (Morone saxatilis W.). We conducted laboratory experiments (feeding and non-feeding) to examine behavioral interactions between adult bluefish and sub-adult striped bass in a large (121,000 L) research aquarium. Additionally, we examined diet and habitat overlap of bluefish and striped bass from the fall and spring bottom trawl surveys conducted by the National Marine Fisheries Service. Observations of feeding trials for the following treatments were made: non-impaired (i.e., same number and size of bluefish and striped bass), size-impaired (i.e., large striped bass/small bluefish), number-impaired (i.e.,10 striped bass/3bluefish), and single-species controls. Within a species, there was no difference in a variety of behavioral measures (e.g., attack rate, capture success, ingestion rate, and activity) between mixed- and control treatments under non-impaired or size-impaired conditions. However, behavior of number-impaired bluefish differed from control and size-impaired fish suggesting that striped bass may have a negative influence on bluefish foraging when bluefish are “out-numbered”. Feeding had a strong effect on swimming speeds for both species. Diet and habitat overlap between bluefish and striped bass in continental shelf waters was low. Overall, foraging behavior in mixed-species treatments and field observations suggest no competitive interactions between adult bluefish and sub-adult striped bass.     

Comparative study of proteolytic enzymes in the digestive tracts of the european sea bass and hybrid striped bass reared in freshwater

• 1.1. A comparative study of the proteolytic activity in four different sections of the digestive tracts of the European sea bass (Dicentrarchus labrax) and hybrid striped bass (Morone chrysops × M. saxatilis) reared in freshwater revealed minor differences between these fish.
• 2.2. Tryptic activity plays a major role in the proteolytic process in both fish.
• 3.3. The activity of seven intestinal proteolytic enzymes was detected utilizing a combination of specific substrates and inhibitors.
• 4.4. High levels of proteolytic activity were detected in both the proximal and distal sections of the fish intestine at a high pH range (9–10).
• 5.5. In situ monitoring of pH levels revealed a lower pH level in the intestinal proximal section of hybrid striped bass compared with the distal section.
• 6.6. In contrast, higher pH levels were detected at the proximal compared with the distal sections of D. labrax intestine.

     

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.     

Sequence conservation among fish myostatin orthologues and the characterization of two additional cDNA clones from Morone saxatilis and Morone americana

Myostatin (MSTN) negatively regulates mammalian skeletal muscle growth and development by inhibiting myoblast proliferation. Mice and cattle possessing mutant MSTN alleles display a 'double muscling' phenotype characterized by extreme skeletal muscle hypertrophy and/or hyperplasia. MSTN orthologues have been previously characterized in 12 vertebrate species, including the white bass Morone chrysops. Presented herein is the identification and characterization of novel cDNA clones from two additional Morone species: saxatilis (striped bass) and americana (white perch), which were obtained by PCR amplification and subsequent TA-cloning. The predicted amino acid sequence of each cDNA clone contains a putative signal sequence, conserved cysteine residues and a RXXR proteolytic processing site. The different Morone proteins were 97-99% identical to each other and approximately 91, 81, 68 and 67% identical to the tilapia, zebrafish, mammalian and avian proteins, respectively. However, the bioactive domains, which lie downstream of each processing site, were considerably more conserved. They were 99-100% identical within the genus and were approximately 99, 95, 88 and 88% identical to the tilapia, zebrafish, mammalian and avian domains, respectively. This high level of sequence conservation among all known MSTN orthologues suggests that the structure/function relationship of each is equally well conserved among vertebrates.