Population Structure and Variation in Red Snapper (<Emphasis Type="Italic">Lutjanus campechanus</Emphasis>) from the Gulf of Mexico and Atlantic Coast of Florida as Determined from Mitochondrial DNA Control Region Sequence

The mitochondrial DNA control regions of red snapper (Lutjanus campechanus) from the Gulf of Mexico (n = 140) and Atlantic coast of Florida (n = 35) were sequenced to generate a prestocking genetic baseline for planned stock enhancement. Intrasample haplotype and nucleotide diversities ranged from 0.94 to 1.00 and 1.8% to 2.5%, respectively. All population analyses were consistent with the hypothesis that red snapper constitute a single, panmictic population over the sampled range. A ubiquitous, predominant haplotype, shared by 23% of the specimens, appeared to be evolutionarily recent, in contrast to previous findings based on restriction fragment length polymorphism data. Tajimas D values were suggestive of a recent bottleneck. Mismatch distributions from Gulf samples were smooth and unimodal, characteristic of recent population expansion. However, the Atlantic sample exhibited a comparatively broader, possibly multimodal distribution, suggestive of a more stable population history. Additional control-region data may clarify potentially disparate demographic histories of Gulf and Atlantic snapper.     

Diel spawning periodicity of red snapper Lutjanus campechanus in the northern Gulf of Mexico

Ovaries of red snapper Lutjanus campechanus were examined histologically to determine rates of oocyte maturation, diel spawning periodicity and whether lunar cycle influenced spawning rhythm. Hydration of red snapper oocytes began during the mid‐morning hours; c . 5 h was necessary for oocytes to become fully hydrated and ovulation occurred no more than 5 h after oocytes attained full hydration. Appearance of fresh postovulatory follicles after 1330 hours and the absence of hydrated oocytes after 1830 hours signified that red snapper spawning occurred during this 5 h period. In addition, evidence of a peak in spawning was seen near 1600 hours. Postovulatory follicles degenerated within a 24 h time period. A lunar spawning cycle was not evident.     

Isolation and characterization of microsatellites in lane snapper (Lutjanus synagris), mutton snapper (Lutjanus analis), and yellowtail snapper (Ocyurus chrysurus)

Polymerase chain reaction primer pairs for a total of 25 nuclear‐encoded microsatellites (loci) were developed from genomic DNA libraries of lane snapper (Lutjanus synagris), mutton snapper (Lutjanus analis), and yellowtail snapper (Ocyurus chrysurus). The microsatellites include 24 perfect (21 dinucleotide and three trinucleotide) and one imperfect (combination tetranucleotide/tetranucleotide) repeat motifs. A total of 32 individuals of each species were assayed for allelic variation at all 25 microsatellites; reliable amplification products were generated for lane snapper (25 loci), mutton snapper (21 loci), and yellowtail snapper (24 loci). Significant deviations from Hardy–Weinberg expectations, following Bonferroni corrections, were found for one microsatellite in lane and yellowtail snappers, and for three microsatellites in mutton snapper. All pairwise comparisons of microsatellites (all three species) did not deviate significantly from genotypic equilibrium.     

Protection of red snapper (Lutjanus sanguineus) against Vibrio alginolyticus with a DNA vaccine containing flagellin flaA gene

Aims: The main aims of this study were to construct a DNA vaccine containing flagellin flaA gene from Vibrio alginolyticus strain HY9901 and to explore the potential application of pcDNA‐flaA as a DNA vaccine candidate for red snapper (Lutjanus sanguineus). Methods and Results: Plasmid DNA encoding flagellin flaA gene (designated as pcDNA‐flaA) was used as a DNA vaccine to immunize red snapper. The distribution, expression and immunoprotection of the DNA vaccine were analysed in tissues of the red snapper by PCR, RT‐PCR and challenge test. PCR results indicated that pcDNA‐flaA distributed in liver, spleen, kidney, gill and injection site muscle at 7–28 days after vaccination. RT‐PCR results indicated that the flaA gene was expressed in all above tissues of vaccinated fish at 7–28 days after vaccination. In addition, fish receiving the DNA vaccine developed a protective response to live V. alginolyticus challenge 28 days post inoculation, the relative per cent survival (RPS) was 88%. Conclusions: This study showed that injection of pcDNA‐flaA induced an efficient, systemic and antigen‐specific immune response in red snapper, which makes it an effective vaccine candidate against V. alginolyticus infection. Significance and Impact of the Study: The finding that red snapper does adequately respond to pcDNA‐flaA intramuscular injection makes pcDNA‐flaA a promising candidate for DNA vaccine treatment. Furthermore, the availability of red snapper for foreign gene expression represents a useful model to develop effective prophylactic strategies and opens new perspectives for the treatment of bacterial pathogens of marine cultured fish.     

Isolation and characterization of microsatellite DNA loci from Russell's snapper (Lutjanus russellii)

A total of 37 microsatellite loci from the Russell's snapper, Lutjanus russellii, were successfully isolated and characterized. Thirty‐four loci were polymorphic in L. russellii samples. Twenty of the 37 markers did not differ significantly from Hardy–Weinberg expected genotype proportions. Significant linkage disequilibrium was detected in one pairwise comparison. The numbers of alleles and observed heterozygosities in polymorphic loci ranged from two to 16 and from 0.41 to 0.95, respectively. These markers will be useful for studying the population genetic structure of this species.     

Isolation and characterization of microsatellite loci from mangrove red snapper Lutjanus argentimaculatus

Lutjanus argentimaculatus, also called mangrove red snapper, is a commercially important fish in East Asia. A proper understanding of population structure is primarily linked with the management of genetic resources in exploiting marine fisheries. Herein, seven microsatellite loci, which showed high polymorphism (observed heterozygosity per locus ranging from 0.3571 to 0.7857 and expected heterozygosity per locus ranging from 0.6236 to 0.8821), were isolated and characterized from L. argentimaculatus. Cross‐species amplifications also indicate that primers designed for these loci may be useful for further studies about other closely phylogenetic species of the family Lutjanidae.     

Microsatellite and mitochondrial DNA analyses of the genetic structure of blacktip shark (Carcharhinus limbatus) nurseries in the northwestern Atlantic, Gulf of Mexico, and Caribbean Sea

Abstract We investigated the genetic structure of blacktip shark (Carcharhinus limbatus) continental nurseries in the northwestern Atlantic Ocean, Gulf of Mexico, and Caribbean Sea using mitochondrial DNA control region sequences and eight nuclear microsatellite loci scored in neonate and young-of-the-year sharks. Significant structure was detected with both markers among nine nurseries (mitochondrial PhiST = 0.350, P < 0.001; nuclear PhiST = 0.007, P < 0.001) and sharks from the northwestern Atlantic, eastern Gulf of Mexico, western Gulf of Mexico, northern Yucatan, and Belize possessed significantly different mitochondrial DNA haplotype frequencies. Microsatellite differentiation was limited to comparisons involving northern Yucatan and Belize sharks with nuclear genetic homogeneity throughout the eastern Gulf of Mexico, western Gulf of Mexico, and northwestern Atlantic. Differences in the magnitude of maternal vs. biparental genetic differentiation support female philopatry to northwestern Atlantic, Gulf of Mexico, and Caribbean Sea natal nursery regions with higher levels of male-mediated gene flow. Philopatry has produced multiple reproductive stocks of this commercially important shark species throughout the range of this study.     

Multiplex genotyping of novel tetranucleotide microsatellites from a marine foodfish species crimson red snapper (Lutjanus erythropterus)

Crimson red snapper (Lutjanus erythropterus) is an important marine foodfish species in Asia with great potentials for aquaculture. We isolated nine polymorphic microsatellites, among which eight were tetranucleotide repeats, and one was CA‐microsatellite. The average allele number present in 48 individuals was 11.1/locus, ranging from three to 33. The expected heterozygosity ranged from 0.49 to 0.97 with an average of 0.74. Six of the nine markers conformed to the Hardy–Weinberg expectations. No pairwise markers showed the possibility of linkage. Protocols for two multiplex polymerase chain reactions (PCRs), each amplifying two and seven markers, respectively, were presented. The developed microsatellites and optimized multiplex PCRs will be useful for studying population structure of wild stocks and parentage assignment for cultured populations.     

Morphological and biochemical effects of food deprivation during the early development of Pacific red snapper Lutjanus peru

We report the effects of food deprivation on the early development of Pacific red snapper Lutjanus peru during the first days of development. The point of no return (PNR) was determined using the feeding incidence after a delay in first feeding. The gradual deterioration of the larvae during food deprivation was recorded using morphometric, histological, enzymatic and biochemical analysis. The time to reach the PNR was 120 h after hatching. Morphologically, the total length, muscle height, head length, tail length and pectoral angle showed the biggest reductions and their growth coefficients changed significantly during food deprivation. Histologically, enterocyte height also was reduced significantly. The protein concentration and activities of the digestive enzymes trypsin, cathepsin-like and lipase showed a significant decrease; meanwhile, amylase activity remained constant during food deprivation. The concentration of total essential free amino acids (EFAAs) decreased significantly while that of the nonessential free amino acids (NEFAAs) remain stable during food deprivation. The most abundant EFAAs were lysine, leucine, isoleucine and valine; the most abundant NEFAAs were alanine, glycine and glutamate, suggesting a more prominent role as energy substrates. At the time of the PNR the concentration of almost all the free amino acids showed a significant decrease. Early food deprivation has a significant impact on the morphology and biochemical characteristics of L. peru. These results suggest that initial feeding of L. peru should begin within 3 days of yolk sac depletion to avoid the PNR. Further studies are necessary to confirm and validate the characters identified in this study as biomarkers of starvation under culture conditions and evaluate their possible utility in ichthyoplankton surveys.     

Isolation and Characterization of Four Microsatellite Loci in the Tidewater Goby (Eucyclogobius newberryi)

The first microsatellite loci for the endangered tidewater goby Eucyclogobius newberryi are described. A partial tidewater goby genomic library was constructed in pUC19 plasmid. Microsatellite-containing clones were isolated, sequenced, and amplified. Out of 12 positive clones sequenced, 3 were polymorphic and proved useful in a preliminary genetic screen of 3 well-separated populations (50 individuals each) along the central California coast. The number of alleles per locus ranged from 2 to 3. This initial study suggests a population bottleneck had occurred in one of the populations. Received May 30, 2000; accepted September 14, 2000.     

Evidence of multiple vicariance in a marine suture‐zone in the Gulf of Mexico

Aim Our aim was to assess whether single or multiple vicariance event(s) have occurred in a marine suture‐zone in the northern Gulf of Mexico. We estimated the divergence time of two genetically distinct groups of lane snapper (Lutjanus synagris), distributed on either side of this zone, and compared it to the timing of other vicariance events described previously in the area. Location The northern Gulf of Mexico from the Texas coastline to the Atlantic coast of South Florida, USA. The marine suture‐zone is proximal to Mobile Bay in Alabama. Methods A Bayesian Markov chain Monte Carlo (MCMC) approach was used to estimate mutation‐corrected divergence and genetic migration between eastern and western groups of lane snapper, based on genotypic differences at 13 nuclear‐encoded microsatellites obtained previously from 77 and 171 individual samples, respectively. A second estimate of divergence time for eastern and western groups of lane snapper was generated, based on 590 base pairs of DNA sequence from the mitochondrially encoded NADH dehydrogenase subunit 4 (ND4) obtained previously from 45 and 93 individual samples, respectively. Results Both classes of genetic markers (microsatellites and mtDNA) indicated a fairly recent divergence. Confidence intervals for microsatellite‐based estimates suggested that divergence began less than 21 thousand years ago (ka), while confidence intervals for mitochondrial DNA‐based estimates suggested divergence began less than 130 ka. These estimates were not consistent with vicariance events in the literature, which are hypothesized to have occurred well before 135 ka. Main conclusions These results indicate that the marine suture‐zone in the northern Gulf of Mexico features multiple vicariance events and may be characterized by a complex geological/environmental history. We suggest that processes during or after the Wisconsin glaciation (c. 110–18 ka) may have created a previously unrecognized barrier for lane snapper and possibly other species as well.     

Microsatellite loci from red sea urchins (Strongylocentrotus franciscanus)

Strongylocentrotus franciscanus is a keystone marine herbivore in the Pacific North West of North America, that is commercially harvested for its roe in wild fisheries and is under development as an aquaculture species. We report the development of 11 novel, highly polymorphic di‐ and trinucleotide loci for use in population studies of this species. In a survey of over 200 individuals from three coastal sites in British Columbia, observed heterozygosities of the loci ranged from 0.39 to 0.85 and an average of 26 alleles per locus were resolved. Seven of the 11 microsatellite loci were in Hardy–Weinburg equiulibruim (HWE).     

Population Genetic Structure of Pacific White Shrimp (<Emphasis Type="Italic">Litopenaeus vannamei</Emphasis>) from Mexico to Panama: Microsatellite DNA Variation

Genetic variation and population structure of wild white shrimp (Litopenaeus vannamei) from 4 geographic locations from Mexico to Panama were investigated using 5 microsatellite DNA loci. The genetic diversity between populations was indicated by the mean number of alleles per locus and mean observed heterozygosity, which ranged from 7.4 to 8.6 and from 0.241 to 0.388, respectively. Significant departures from Hardy-Weinberg equilibrium were found at most locations at each locus, with the exception Guatemala at Pvan0013, and were caused by high heterozygote deficiencies. Genetic differences between localities were detected by pairwise comparison based on allelic and genotypic frequencies, with the exception of locus Pvan1003. Significant pairwise F _ST values between locations and total F _ST showed that the white shrimp population is structured into subpopulations. However, population differentiation does not follow an isolation-by-distance model. Knowledge of the genetic diversity and structure of L.vannamei populations will be of interest for aquaculture and fisheries management to utilize and preserve aquatic biodiversity.     

Allozyme differentiation within and between red drum (Sciaenops ocellatus) from the Gulf of Mexico and Atlantic Ocean

Nine polymorphic nuclear-gene (allozyme) loci were surveyed among 491 red drum ( Sciaenops ocellatus ) sampled in 1988 and 1989 from nearshore localities in the northern Gulf of Mexico (Gulf) and the Atlantic coast of the southeastern United States (Atlantic). Data were combined with those from a previous study to generate a data set of 762 individuals representing 11 sample localities in the Gulf and 175 individuals representing five sample localities in the Atlantic. The combined data set included individuals from the 1986 and 1987 year classes and permitted rigorous testing of both temporal and spatial genetic heterogeneity. Average heterozygosity-per-locus values (estimated using 33 assumed monomorphic loci) were 0·048 (Gulf red drum) and 0·046 (Atlantic red drum). Tests of heterogeneity in allele frequencies between year classes at individual localities and across regions (Gulf and Atlantic) were non-significant. Tests of spatial (geographic) heterogeneity indicated that red drum are weakly subdivided: genetically-differentiated subpopulations occur in the northern Gulf and along the south-eastern Atlantic coast. Genetic data were consistent with the hypothesis that red drum within the Gulf and along the Atlantic coast comprise singie subpopulalions. Genetic differences between Gulf and Atlantic red drum seem likeiy to stem from historical or recent interactions between dispersal and impediments to gene flow.     

Interactions between range‐expanding tropical fishes and the northern Gulf of Mexico red snapper Lutjanus campechanus

Experimental investigation of the intensity of potential competitive interactions among increasingly abundant tropically‐associated grey Lutjanus griseus and lane snapper Lutjanus synagris and resident northern Gulf of Mexico (nGOM) red snapper Lutjanus campechanus was undertaken in large outdoor mesocosms. In pair‐wise interaction trials, compared with L. synagris, L. campechanus demonstrated significantly increased roving behaviour and predatory activity. While no significant difference in these activities was observed between L. campechanus and L. griseus, when all three snappers (Lutjanidae) were grouped together L. campechanus swimming activity significantly decreased in the presence of both tropically‐associated species. Overall, L. campechanus were more active and aggressive predators and appear to be competitively resistant to L. griseus and L. synagris. As lower latitude species have continued to become increasingly prevalent in nGOM habitats and regional warming continues to affect resident reef‐associated fishes, these findings contribute to the assessment of the effects of warming‐related species shifts upon nGOM fishes and document current partial resilience of L. campechanus to climate‐related expansions of tropical confamilials.     

Vibrios of the spotted rose snapper Lutjanus guttatus Steindachner, 1869 from northwestern Mexico

AIM: To characterize and identify vibrios present in wild and cultured juvenile snappers (Lutjanus guttatus) in northwestern Mexico. METHODS AND RESULTS: Spotted rose snapper juveniles were collected at four localities in northwestern Mexico. Bacteria were isolated from external lesions, kidney, liver, and spleen from cultured and wild caught organisms. In total, 280 isolates were obtained and fingerprinted with rep-PCR (GTG5). Nearly 93.2% of the strains belonged to the Vibrionaceae family. Species and genera identified were Photobacterium damselae subsp. damselae (76 strains), Photobacterium leiognathi (13), Vibrio sp. (24), Vibrio alginolyticus (12), Vibrio campbellii (19), Vibrio fortis (9), Vibrio harveyi (49), Vibrio ichthyoenteri (4), Vibrio mediterranei (4), Vibrio parahaemolyticus (1), Vibrio ponticus (2), Vibrio rotiferianus (22), and four potential new species. Conclusions: A wide diversity of vibrios was found in the external lesions and internal organs of both wild and cultured spotted rose snapper juveniles. In total, 12 species of vibrios and four potential new species were identified. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study on the vibrios present in the spotted rose snapper and therefore might serve as a basis for future studies and diagnosis.     

Isolation and Characterization of Twelve Microsatellite Loci for Rockfish (Sebastes)

We describe the first microsatellites for rockfishes in the diverse genus Sebastes. Clones containing microsatellites were isolated from the genomic library of a quillback rockfish, Sebastes maliger. Twelve microsatellites are characterized; six of these are polymorphic in quillback rockfish, and eight are polymorphic in at least one rockfish species on which they were tested. The number of alleles per variable locus ranged from 4 to 15 and averaged 6.8. The expected heterozygosities ranged from 0.38 to 0.79 and averaged 0.60 in these loci. These loci should prove valuable in studies examining species identification, population genetics, hybridization, paternity, kinship, and microsatellite evolution. Received September 8, 1998; accepted November 23, 1998.