Estimation of effective population size for the long-lived darkblotched rockfish Sebastes crameri

We report the variance effective population size (Ne) in darkblotched rockfish (Sebastes crameri) utilizing the temporal method for overlapping generations, which requires a combination of age-specific demography and genetic information from cohorts. Following calculations of age-specific survival and reproductive success from fishery data, we genotyped a sample (n = 1087) comprised by 6 cohorts (from 1995 to 2000) across 7 microsatellite loci. Our Ne estimate (Ne) plus 95% confidence interval was (Ne) = 9157 [6495-12 215], showing that the breeding population number could be 3-4 orders of magnitude smaller than the census population size (N) = 24 376 210). Our estimates resemble closely those found for fishes with similar life history, suggesting that the small (Ne)/(N) ratio for S. crameri is most likely explained by a combination of high variance in reproductive success among individuals, genetic structure, and demographic perturbations such as historical fishing. Because small (Ne)/(N) ratios have been commonly associated with potential loss of genetic variation, our estimates need careful consideration in rockfish management and conservation.     

Comparative allometric growth of the gastrointestinal tract of two sympatric congeners, copper rockfish (Sebastes caurinus) and quillback rockfish (S. maliger)

Allometric growth of the stomach, intestine, and caeca was compared between two sympatric congeners, the copper rockfish ( Sebastes caurinus ) and the quillback rockfish ( S. maliger ), to examine the species-specific differences in their gut tract in relation to their general feeding habits. The intestine of copper rockfish was shorter and its mass less than the intestine of quillback rockfish. Copper rockfish also had a greater stomach mass and, on average, a greater number of pyloric caeca than quillback rockfish. The differences in the gut allometry between the species were consistent with differences in their general food habits as copper rockfish consume proportionally more fish and larger crustaceans than quillback rockfish.     

Gastric evacuation rates for two foods in the black rockfish, Sebastes melanops Girard

The rates of gastric evacuation of two food types, squid and fish, were measured in the laboratory for adult black rockfish, Sebastes melanops , held nominally at 11°C. Linear, logarithmic and square root regression models were applied to analyse the data expressed as both wet and dry weight proportions of the original meal. The linear model provided the best fit for the wet weight relationship of squid. The dry weight of squid and both the wet and dry weight of fish were best described by a logarithmic model, or by a square root model when the regressions were' forced' through the original meal size. The instantaneous evacuation rates (≃ 6% h⁻¹) and times to complete evacuation (≃ 76 h) were similar for all relations except those involving the wet weight of squid. The addition of the meal size as an independent variable improved the models in most cases whereas the addition of the weight of fish used in the experiments did not improve the model in any instance.     

Reproductive performance of yellowtail rockfish,Sebastes flavidus

Synopsis Field and laboratory studies were conducted for 3 years on the yellowtail rockfish,Sebastes flavidus, from Cordell Bank, California, in order to characterize the reproduction of this species whose northern stocks have declined. Research findings included reversal of the sex ratios and male-female ages and sizes at age throughout the annual cycle, heavier and longer, females at age than males after sexual maturation, maturation of females at 6 and males at 8 years, long reproductive lifespans, distinct male and female gonadosomatic index patterns over the annual cycle, age- and size-specific fecundity, no difference between potential and realized fecundity and the seasonal changes associated with gonadogenesis. The reproductive profile of the Cordell Bank yellowtail rockfish provided a base for comparison with northern populations that appeared to differ, especially in age and size.     

Metabolic costs of viviparity in yellowtail rockfish,Sebastes flavidus

Synopsis We estimated the energy utilized by developing embryos and larvae of yellowtail rockfish,Sebastes flavidus, by determining the routine metabolic rate of adult females throughout gestation and subtracting the mean metabolic rate of non-gestating fish. The pattern of maternal oxygen consumption during gestation was such that metabolic demands did not become significantly elevated until intra-ovarian embryos had hatched and larvae were being incubated (i.e. 24–28 d post-fertilization). Metabolic rates of females in this latter stage of gestation were 82% higher than spent females and 101% higher than those of non-reproductive males. We determined that 0.08996 cal embryo^-1 was consumed during gestation as compared to the change in energy content of embryos from fertilization to birth which was 0.08690 cal embryo^-1. The difference between the energy consumed and the energy lost during the 29 d gestation period indicated that extra embryonic sources contributed only 3.4% or 0.00306 cal embryo^-1 of the energy utilized after fertilization. Our observations agree with previous work which determined that embryos and larvae ofS. flavidus are primarily lecithotrophic and that the relatively high fecundity ofS. flavidus coupled with a small egg size, rapid embryogenesis and brief gestation period characterize this species as a relative r-strategist within a genus whose species range broadly over the K end of the r-K continuum.     

Molecular characterization, expression, and functional analysis of two thioredoxins in the black rockfish (Sebastes schlegelii)

Thioredoxins (TRxs) are a family of small evolutionarily conserved proteins that are essential for the maintenance of cellular homeostasis. Two TRx homologue cDNAs were isolated from a black rockfish concanavalin A (Con A)/phorbol myristate acetate (PMA)-stimulated leucocyte cDNA library and named BrTPx1-1 and BrTPx1-2. As compared with other known TRx peptide sequences, the most conserved regions of both BrTRx1-1 and BrTRx1-2 peptides were found to be the redox-active site Trp-Cys-X-X-Cys (WCXXC). The TRx present in most species is a TRx1-2 protein with a Cys-Pro-Gly-Cys (CPGC) active site. However, in the larger 13 kDa BrTRx1-1 protein, a Cys-Pro-Pro-Cys (CPPC) active site was identified. Here, we report the identification of a new member of the TRx protein family from the teleost black rockfish, which defines a new subclass of 13-kDa TRx1-1 proteins. Phylogenetic analysis indicated that both BrTRx1-1 and BrTRx1-2 were grouped with other vertebrate TRx1 peptides. BrTRx1-1 expression was strongly induced in peripheral blood leucocytes (PBLs) 12-24 h following Con A/PMA stimulation, with peak expression at 24 h post-stimulation. BrTRx1-2 was induced in PBLs after stimulation with lipopolysaccharide (LPS), Con A/PMA, or poly I:C at 24 h. The BrTRx1-1 gene was predominantly expressed in the liver and gills, while BrTRx1-2 was expressed in PBLs and gills. After treatment with a high concentration (10 μg/mL) of rBrTRx1-1 or rBrTRx1-2, kidney leucocytes exhibited increased cell proliferation and viability under oxidative stress.     

Cloning and characterization of novel microsatellite DNA markers for the grass rockfish,Sebastes rastrelliger,and cross‐species amplification in 10 related Sebastes spp

Here we report development and characterization of seven polymorphic loci derived from grass rockfish (Sebastes rastrelliger) genomic DNA phagemid libraries enriched for microsatellite motifs. Within grass rockfish, average allelic diversity was 11.3 alleles per locus and average heterozygosity was 0.73. The seven loci also were surveyed in 10 related species of Sebastes, where allelic diversity ranged from highly polymorphic to monomorphic. Deviations from Hardy–Weinberg were nonsignificant in all but one species/locus combination suggesting low occurrence of null alleles. Significant linkage disequilibrium was detected among three loci, but these events were limited to a single species in each case.     

Microsatellite markers for the heavily exploited canary (Sebastes pinniger) and other rockfish species

The isolation and characterization of nine polymorphic microsatellite loci (eight tetranucleotide and one dinucleotide) from the canary rockfish Sebastes pinniger are described. Polymorphism at these loci revealed from six to 28 alleles, with expected heterozygosities ranging between 0.42 and 0.88, enabling high‐resolution genetic population structure investigation for this overfished species in the northeastern Pacific. They also amplify in 13 other congeneric species, providing highly variable loci for research on other rockfishes.     

Isolation and characterization of microsatellite markers for the heavily exploited rockfish Sebastes schlegeli, and cross-species amplification in four related Sebastes spp.

Here we report development and characterization of 14 polymorphic microsatellite loci from Sebastes schlegel. Polymorphism at these loci revealed from 3 to 23 alleles. The observed heterozygosity ranged from 0.34 to 1.00, and the expected heterozygosity ranged from 0.31 to 0.95. No linkage disequibrium was found. Two loci were significantly deviated from HWE (P < 0.01). The 14 loci were also surveyed in four other Sebastes species and 12 loci successfully amplified, where allelic diversity ranged from highly polymorphic to monomorphic. These results demonstrate these microsatellite markers can be used for the study of intra- and inter-specific genetic diversity.     

Isolation and characterization of microsatellite markers for the Korean rockfish, Sebastes schlegeli

The Korean rockfish (Sebastes schlegeli) is an important commercial fish that is widely used in aquaculture. We isolated and characterized 18 polymorphic microsatellite loci from the Korean rockfish using a (GT)(13)-enriched genomic library. Polymorphism was assessed in 48 individuals from a single population collected from the northern coastal waters of the Yellow Sea. The observed and expected heterozygosities ranged from 0.0244 to 0.7660 (mean 0.4194) and 0.0244 to 0.8758 (mean 0.5002), respectively. Polymorphism at these loci indicated from two to 15 alleles (mean 5.7); 14 of 18 loci conformed to Hardy-Weinberg equilibrium. These markers should be useful for management and conservation studies of this species.     

Henneguya sebasta sp. n. (protozoa, myxosporida) from California rockfish, Sebastes spp.

Henneguya sebasta sp. n. was found on the bulbus and truncus arteriosus and in the heart chambers of 7 species of marine rockfish, Sebastes, from central and southern California. The incidence of this parasite may be of economic interest to the sport and commercial fisheries because of its possible pathogenicity.     

Purification and characterization of luteinizing hormone from pituitary glands of rockfish, Sebastes schlegeli

To acquire greater knowledge of the reproductive function of luteinizing hormone (LH) in the viviparous rockfish Sebastes schlegeli, LH from the pituitary glands of mature rockfish was isolated, purified, and localized and its biological activity was characterized. The molecular mass of purified LH was estimated to be approximately 33 kDa, similar to that of known LH. When rockfish LH was purified by reverse-phase high-performance liquid chromatography, its N-terminal amino acid sequences were found to coincide with those of predicted cDNA sequences of rockfish gonadotropin α (ssGTHα) and ssLHβ mature peptides. Immunocytochemical analysis using antisera against ssGTHα (molecular weight [MW], ~14.5 kDa) and ssLHβ (MW, ~18.5 kDa) indicated that the LH-producing cells are mainly distributed throughout the proximal pars distalis and along the periphery of the pars intermedia. Further, in vitro ovarian follicle analysis demonstrated that purified intact rockfish LH significantly enhances E(2) secretion in a dose-dependent manner. This is the first report on the purification and characterization of LH from a viviparous teleost, and these results will enable future research and increase our understanding of the mechanisms underlying the maturation of such fish.     

Energy utilization by embryos during gestation in viviparous copper rockfish,Sebastes caurinus

Synopsis The energy utilized by developing copper rockfishSebastes caurinus embryos was measured through direct respirometry (0.408 cal embryo⁻¹) and compared with the change in energy content of embryos from fertilization to birth (0.361 cal embryo⁻¹). The difference between the energy consumed and energy lost during gestation indicates that 0.047 cal embryo⁻¹ or 11.5% of the energy utilized during gestation is contributed by the mother after fertilization. Available information on other membres of the genusSebastes suggests that matrotrophic viviparity may be the common mode of reproduction, but energy contribution varies greatly between species.     

Immunochemical identification and partial characterization of female-specific serum proteins in white-edged rockfish,Sebastes taczanowskii

Synopsis Female-specific serum proteins (FSSPs) in white-edged rockfish,Sebastes taczanowskii, were identified and partially characterized by immunochemical procedures. Two FSSPs, which clearly reacted with antiserum against egg proteins, were confirmed in the serum of mature females, and estrogen treatment induced similar FSSPs in the serum of mature males. Hence, the FSSPs were considered to be vitellogenin. The vitellogenin concentration in female fish was high during the vitellogenic period and low during gestation, parturition and the recovery period, indicating that vitellogenin is used only for yolk formation in the oocytes and not as a direct nutritional source for developing embryos during gestation. On the other hand, an FSSP (FS3), which was considered not to be vitellogenin, was also identified in the sera of mature females and males after estradiol-17β administration by using an antiserum (a-FS3) that removed the components of the male serum and egg extracts from the anti-mature female serum antiserum. Moreover, immunohistochemical observation with a-FS3 illustrated that FS3 was a major constituent of the ovarian fluid but not of vitellogenic oocytes. The cross-reactivities of these FSSPs among seven viviparous rockfishes demonstrated that vitellogenin existed in the sera of all rockfishes studied belonging to the generaSebastes andSebastiscus, whereas FS3 was not present in several species ofSebastes.     

Molecular cloning and characterization of peptidoglycan recognition proteins from the rockfish,Sebastes schlegeli

Peptidoglycan recognition proteins (PGRPs) are innate immune molecules that are structurally conserved through evolution in both invertebrate and vertebrate animals. Here we report the identification and characterization of two long forms of PGRP (SsPGRP-L1 and SsPGRP-L2) from the rockfish, Sebastes schlegeli. The deduced amino acid sequences of SsPGRP-L1 and SsPGRP-L2, 466 and 482 residues respectively, contain the conserved PGRP domain and the four Zn²⁺-binding amino acid residues required for amidase activity. In addition to peptidoglycan-lytic amidase activity, recombinant SsPGRPs have broad-spectrum antimicrobial activity like zebrafish PGRPs. RT-PCR analysis of total RNA shows that the expression patterns of SsPGRP-L1 and SsPGRP-L2 genes are different, though they are widely expressed in the tissues that come in contact with bacteria. Overall, these data suggest that rockfish PGRPs are involved in the innate host defense of S. schlegeli against bacterial infections.