Integrating fish and parasite data as a holistic solution for identifying the elusive stock structure of Pacific sardines (Sardinops sagax)

There is an urgent need to clarify how different stocks, or subpopulations of fish species, are vulnerable to fishing pressure and unfavorable ocean conditions because of the increasing demand on fisheries for human consumption. For marine fishes, the potential for high gene flow increases the difficulty in determining the number of subpopulations managed in a specific fishery. Although the use of molecular data has become a common method in the past 15 years to identify fish subpopulations, no single technique or suite of techniques has been established for fish stock structure studies. We review the use of fish morphometrics, artificial tags, fish genetics, parasite genetics, and parasites as biological tags to identify subpopulations of marine fishes with a focus on the Pacific sardine (Sardinops sagax) fishery off the west coast of North America. We suggest an integration of fish- and parasite-based techniques for future stock structure studies, particularly for pelagic fish species whose stock structure can be elusive. An integration of techniques may also resolve fish stock structure over small geographic areas by increasing the number of spatial and temporal scales studied simultaneously leading to methods for successful management of marine fish species.     

Characterization of polymorphic microsatellites in the Pacific sardine Sardinops sagax sagax (Clupeidae)

We isolated 11 microsatellites from the Pacific sardine Sardinops sagax sagax. The number of alleles and H_E among 30 individuals from a single population ranged from four to 24 and from 0.606 to 0.959, respectively. Pacific sardines are a vital economic resource in the northeastern Pacific Ocean, but insufficiently polymorphic loci have limited inferences about its stock structure and genetic variation. The level of variability of these new markers will prove useful in testing hypotheses on the stock‐structure and long‐term genetic integrity of the species.     

Population genetic structure of three tree species in the mangrove genus Ceriops (Rhizophoraceae) from the Indo West Pacific

Ceriops is a viviparous mangrove with widespread species Ceriops decandra and C. tagal, and an endemic species C. australis. Genetic diversity of the three species was screened in 30 populations collected from 23 locations in the Indo West Pacific (IWP) using Inter-simple sequence repeats (ISSR) and sequences of partial nuclear gene (G3pdh) and chloroplast DNA (trnV-trnM). At the species level, the total gene diversity (Ht) revealed by ISSRs was 0.270, 0.118, and 0.089 in C. decandra, C. tagal, and C. australis, respectively. A total of six haplotypes of G3pdh and five haplotypes of trnV-trnM were recognized among the three species. Only C. decandra was detected containing more than one haplotype from each sequence data set (four G3pdh haplotypes and three trnV-trnM haplotypes). At the population level, genetic diversity of Ceriops was relatively low inferred from ISSRs (He = 0.028, 0.023, and 0.053 in C. decandra, C. tagal, and C. australis, respectively). No haplotype diversity within population was detected from any of the three species. Cluster analysis based on ISSRs identified three major geographical groups in correspond to the East Indian Ocean (EIO), South China Sea (SCS), and North Australia (NA) in both C. decandra and C. tagal. The cladogram from DNA sequences also detected the same three geographical groups in C. decandra. Analysis of molecular variance (AMOVA) revealed that most of the total variation was accounted for by differentiation between the three major geographical regions of both C. decandra and C. tagal. The significant genetic structure may result from the geological events in these regions during the recent Pleistocene glaciations. This study also provided insights into the phylogenetics of Ceriops. Yelin Huang and Fengxiao Tan contributed equally to this work.     

Changes in growth and maturation parameters of Pacific sardine Sardinops sagax collected off California during a period of stock recovery from 1994 to 2010

Whether fluctuation in density influenced the growth and maturation variables of three aggregated cohorts (fish born during the 1986–1993, 1996–2003 and 2004–2008 periods) of Pacific sardine Sardinops sagax caeruleus collected off the Californian coast from 2004 to 2010 was investigated. Using a von Bertalanffy mixed‐effects model with aggregated cohorts as covariates, estimated growth rate significantly covaried with aggregated cohorts. Growth rate (K) was modelled as a fixed effect and estimated to be 0·264 ± 0·015 (±s.e ). Statistical contrasts among aggregated cohorts showed that the 1996–2003 cohorts had a significantly lower growth rate than the other two aggregated cohorts. The theoretical age at length zero (t₀) and the standard length at infinity (L_S∞) were modelled as random effects, and were estimated to be ?2·885 ± 0·259 (±s.e ) and 273·13 ± 6·533 mm (±s.e ). The relation of ovary‐free mass at length was significantly different among the three aggregated cohorts, with the allometric coefficient estimated to be 2·850 ± 0·013 (±s.e ) for the S. sagax population. The age‐at‐length trajectory of S. sagax born between 1986 and 2008 showed strong density dependence effects on somatic growth rates. In contrast to the density‐dependent nature of growth, the probability to be mature at‐size or at‐age was not significantly affected by aggregated cohort density. The size and the age‐at‐50% maturity were estimated to be 150·92 mm and 0·56 years, respectively. Stock migration, natural fluctuations in biomass and removal of older and larger S. sagax by fishing might have been interplaying factors controlling growth parameters during 1986–2010.     

Living with uncertainty: genetic imprints of climate shifts in East Pacific anchovy (Engraulis mordax) and sardine (Sardinops sagax)

In the upwelling zone of the northeastern Pacific, cold nutrient-rich conditions alternate with warm nutrient-poor intervals on timescales ranging from months to millennia. In this setting, the abundances of Pacific sardine (Sardinops sagax) and northern anchovy (Engraulis mordax) fluctuate by several orders of magnitude, with sardine dominating during warm conditions and anchovy dominating during cool conditions. Two population models can explain the response of these fishes to adverse conditions. Under the basin model, species distributions contract to a central (optimal) range during population crashes. Expectations of this model may include a single range-wide population with a decline in genetic diversity on both sides of a central refuge. In contrast, the self-recruitment model invokes a series of local oceanographic domains that maintain semi-isolated subpopulations. During adverse conditions, some subpopulations cannot complete the life cycle within the local environment and are extirpated. Expectations of this model include some degree of population genetic structure and no clear gradient in genetic diversity. We examined mitochondrial DNA cytochrome b sequences to assess these competing models for anchovy (N = 196; 539 bp) and sardine (N = 107; 425 bp). The mitochondrial DNA gene genealogies are shallow but diverse for both species. Haplotype frequencies are homogeneous among subpopulations, but genetic diversities peak for both species along Baja California and adjacent southern California. Mismatch distributions and Tajima's D-values reveal distinctive signatures of population bottlenecks and expansions. Sardine haplotypes coalesce at approximately 241,000 years bp, with an initial female effective population size Nf0 = 0 followed by exponential growth to Nf1 = 115 million. Anchovy haplotypes coalesce at approximately 282,000 years bp, with an initial population size of Nf0 = 14,000, followed by exponential growth to Nf1 = 2.3 million. These results indicate a founder event for sardine and a severe population decline for anchovy in the California Current during the late Pleistocene. Overall, these data support the basin model on decadal timescales, although local recruitment may dominate on shorter timescales.     

An improved method for studying the condition of fish, with an example using Pacific sardine Sardinops sagax (Jenyns)

The statistical treatment of condition used in many previous studies has been to determine a predicted weight for a fish, and to calculate a ratio of actual to predicted weight. The ratios obtained, known as condition factor K , are then further analysed using analysis of variance or other techniques to relate changes in K to other independent variables. It is suggested that condition can be studied better by using a single model to analyse the response of fish weight to a number of factors simultaneously. This approach affords benefit from the simplicity of using an integrated analysis and avoids problems with skewed distributions of ratios. An example using data on the Pacific sardine, Sardinops sagax (Jenyns), is given. Changes in condition of this fish due to monthly, yearly and temperature effects are calculated.     

Comparative population structure of three snook species (Teleostei: Centropomidae) from the eastern central Pacific

Three snook species, Centropomus viridis, Centropomus medius, and Centropomus robalito, from the eastern central Pacific, representing three of the four proposed phyletic lineages in the genus, were analyzed for genetic variability by means of allozyme and RAPD to evaluate the divergence between populations at different levels of dispersal ability and to evaluate the importance of barriers to dispersal in the population subdivision and genetic diversity. Levels of genetic diversity among species estimated by allozymes were similar and consistent with the observed levels of differentiation in marine fish species. Mean heterozygosity ranged from 0.089 for C. viridis to 0.10 for C. robalito. Genetic diversity for the snook species studied was slightly higher than the mean estimation reported in allozymes for 106 marine fish (0.055) and for anadromous fish species (0.043 to 0.057). Multilocus allele frequency homogeneity tests and population-subdivision estimates for both allozyme and RAPD markers revealed the existence of population structure in C. viridis and C. medius, in coincidence with geographic separation of samples, whereas no divergence was detected in C. robalito. This finding may be attributed to the greater population size of C. robalito, which originated by a recent population range expansion, and hence the potential for dispersal is mediated by larval drift. Fluctuations in population size and population range expansion are used to explain discrepancies between levels of genetic diversity and population structure in the studied species. Supplementary material to this paper is available in electronic format at     

Population genetics of Howellia aquatilis (Campanulaceae) in disjunct locations throughout the Pacific Northwest

Howellia aquatilis A.Gray (water howellia) is a federally-listed threatened aquatic plant species with limited distribution in four states: California, Idaho, Montana, and Washington. Previous studies have shown a lack of genetic variation within the species; these studies, however, have excluded samples from one or more states. There have been no published studies on the population biology or genetics of the six known California populations or their evolutionary relationship to the other Pacific Northwest populations. We used Amplified Fragment Length Polymorphisms to identify genetic variation within and among the California populations, and to compare the California populations to the Idaho, Montana, and Washington populations. Analysis of molecular variance of 92 individuals from the six California populations show that 83.8% of genetic variation is found within populations and 16.2% among populations (P < 0.001). All sampled populations from all states provide 83.7% variation within and 16.3% variation among populations (P < 0.001). A UPGMA analysis confirms there is no clear clustering of Howellia aquatilis populations within California, that the Montana populations cluster within the California populations, and, although with limited population sample sizes, the Idaho and Washington populations are distantly related to all other populations. Waterfowl migration patterns support a hypothesis for avian dispersal as a primary factor in gene flow in Howellia aquatilis.     

Diversity and genetic structure of three species of Dioon Lindl. (Zamiaceae, Cycadales) from the Pacific seaboard of Mexico

We have estimated levels of genetic diversity and partitioning in the Mexican endemic cycad species Dioon sonorense, Dioon tomasellii, and Dioon holmgrenii, whose populations are exclusively distributed along the Pacific seaboard. For the three species, the patterns of variation at 19 allozyme loci in a total of 11 populations were evaluated. The average number of alleles per locus was in the range 2.05–1.68, corresponding to the northernmost population of D. sonorense (Mazatán), and the southernmost population of Dioon holmgrenii (Loxicha), respectively. In turn, the percentage of polymorphic loci peaked (94.73) in the El Higueral and Altamirano populations of Dioon tomasellii, and was estimated to be lowest (57.89) in the Loxicha population of D. holmgrenii. The mean expected heterozygosis varied markedly between taxa, with relatively high indices for D. sonorense and D. tomasellii (H_E = 0.314 and 0.295, respectively) and substantially lower values for D. holmgrenii (H_E = 0.170). Comparison of the inferred genetic structure based on F‐statistics for the three species also indicated differences along the north‐south Pacific seaboard axis. For D. sonorense and D. tomasellii, local inbreeding (F_IS) was zero but global inbreeding (F_IT) values were positive and significantly different from zero (0.130 and 0.116, respectively). By contrast, values of both F_IT and F_IS were negative and significantly different from zero (?0.116 and ?0.201, respectively) for D. holmgrenii. The genetic differentiation between populations (F_ST) had positive values in all taxa and corresponded with their geographic location along the north‐south axis: according to this statistic, D. sonorense was the most differentiated species (F_ST = 0.151), D. tomasellii had intermediate values (F_ST = 0.145), and D. holmgrenii was the less differentiated taxon (F_ST = 0.069). Finally, a phenogram representing Nei's genetic distances among populations displayed three major groups, each one corresponding to each of the studied species. Within D. tomasellii (of intermediate geographic distribution), a further division into two clusters corresponded precisely to the pair of populations that are geographically divided by the Trans Mexican Neovolcanic Mountains. © 2008 The Linnean Society of London, Biological Journal of the Linnean Society, 2008, 94 , 765–776.     

Population structure of Anisakis simplex (Nematoda) in harbor porpoises Phocoena phocoena off Denmark

The population structure and habitat selection of Anisakis simplex in 35 harbor porpoises off Denmark are described. The nematodes were collected from the stomach and duodenal ampulla and were categorized as third-stage larvae, fourth-stage larvae, subadults, and adults. The porpoises harbored 8,043 specimens of A. simplex. The proportion of adults and subadults increased with infrapopulation size. The number of development stages across infrapopulations covaried significantly (Kendall's test of concordance). Concordance was higher in hosts with the highest intensities than in those with low and medium intensities. All stages occurred mainly in the forestomach, but this trend was stronger for the adults. Adult and subadult sex ratios did not depart significantly from 1:1. Our data suggested that recruitment and duration of each stage were the main factors accounting for infrapopulation structure. The preference of A. simplex for the forestomach conformed with previous studies, but the narrower distribution of adults relative to other stages might indicate a strategy to enhance mating opportunities. Information on sex ratios of A. simplex is scarce and contradictory. We suggest that the discrepancies might partly reflect differences in categorization criteria and statistical methods.     

Population structure and taxonomic discrimination among three species of Biomphalaria Preston, 1910 (Gastropoda: Planorbidae) from Kenya

The genetic basis for species designations for African species of Biomphalaria has received little attention. Populations of B. choanomphala, B. sudanica and B. pfeifferi from Kenya were examined using starch gel electrophoresis. Analysis of population structure as measured using Wright's F -statistics for B. sudanica and B. choanomphala revealed that both species show substantial departures from expectations under random mating. Populations of B. choanomphala show the least evidence of inbreeding, and populations of A sudanica are intermediate between B. choanomphala and previously reported levels of inbreeding for B. pfeifferi. Syn topic populations of B. pfeifferi and B. sudanica were genetically distinct. Syntopic populations of B. choanomphala and B. sudanica also showed evidence of separate gene pools, although the evidence is not as conclusive as for B. pfeifferi and B. sudanica. Possible explanations for the failure to find clear differences between B. sudanica and B. choanomphala are considered.     

Population genetic structure of three pond-inhabiting Daphnia species on a regional scale (Flanders, Belgium)

1. Only a few studies have compared patterns of genetic variation among populations of different Daphnia species on a regional scale. The present study addresses this gap and examines the relationship between diversity as revealed by allozyme variation and habitat size for populations of Daphnia pulex, D. obtusa and D. curvirostris in Flanders (Belgium). In addition, we examined whether patterns of isolation‐by‐distance could be observed in each of these three Daphnia species. 2. The relationship between genetic diversity and habitat size varied among Daphnia species that occur in the same region. In D. pulex and D. obtusa populations, a positive relationship between local genetic diversity and habitat size was found, whereas the relationship was negative in D. curvirostris populations. 3. Regional genetic diversity was lower than expected from patterns of local genetic diversity in D. pulex and D. obtusa populations in Flanders. This suggests that the subdivision of local Daphnia populations in a region did not obviously increase genetic diversity. 4. Genetic differentiation among populations of these three species in Flanders was moderate and comparable with values observed in other Daphnia species. Patterns of isolation‐by‐distance could be observed, but the scatter was high (D. pulex) or the slope was very low (D. obtusa).     

Population genetic structure of the catadromous Australian bass from throughout its range

One hatchery-derived impoundment and nine wild riverine populations of the catadromous percoid Australian bass Macquaria novemaculeata were examined at six polymorphic allozyme loci to reveal population genetic structure. Subtle genetic differences among many populations indicated that fish did not interact as a panmictic unit, but rather conformed to an isolation by distance pattern of population structure. The hatchery-derived Glenbawn Dam sample differed from the riverine populations in both average heterozygosity and number of polymorphic loci, demonstrating the need for caution in supplementation programs to rehabilitate wild riverine populations.     

Population genetic structure of three freshwater mussel (Unionidae) species within a small stream system: significant variation at local spatial scales

1. Unionid mussels are highly threatened, but little is known about genetic structure in populations of these organisms. We used allozyme electrophoresis to examine partitioning of genetic variation in three locally abundant and widely distributed species of mussels from a catchment in Ohio. 2. Within‐population variation was similar to that previously reported for freshwater mussels, but genotype frequencies exhibited heterozygote deficiencies in many instances. All three species exhibited significant among‐population variation. Evidence of isolation‐by‐distance was found in Elliptio dilatata and Ptychobranchus fasciolaris, while Lampsilis siliquoidea showed no geographical pattern of among‐population variation. 3. Our results suggest that the isolating effects of genetic drift were greater in L. siliquoidea than in the other species. Differentiation of populations occurred at a much smaller spatial scale than has previously been found in freshwater mussels. Differences among species may reflect differences in the dispersal abilities of fishes that serve as hosts for the glochidia larvae of mussels. 4. Based on our results, we hypothesise that species of mussels that are common to large rivers exhibit relatively large amounts of within‐population genetic variation and little differentiation over large geographical distances. Conversely, species typical of small streams show lower within‐population genetic variation and populations will be more isolated. If this hypothesis can be supported, it may prove useful in the design of conservation strategies that maintain the genetic structure of target species.     

Two new species of phacelia (Hydrophyllaceae) from California

Phacelia cookei andP. stebbinsii, thus far found only in California, are described as new. The former is apparently endemic to ashy volcanic sand on Mt. Shasta; the latter is found on metamorphic rock cliffs in Eldorado County.     

Analysis of the complete plastomes of three species of Membranoptera (Ceramiales,Rhodophyta) from Pacific North America

Next generation sequence data were generated and used to assemble the complete plastomes of the holotype of Membranoptera weeksiae, the neotype (designated here) of M. tenuis, and a specimen examined by Kylin in making the new combination M. platyphylla. The three plastomes were similar in gene content and length and showed high gene synteny to Calliarthron, Grateloupia, Sporolithon, and Vertebrata. Sequence variation in the plastome coding regions were 0.89% between M. weeksiae and M. tenuis, 5.14% between M. weeksiae and M. platyphylla, and 5.18% between M. tenuis and M. platyphylla. We were unable to decipher the complete mitogenomes of the three species due to low coverage and structural problems; however, we assembled and analyzed, the cytochrome oxidase I, II, and III loci and found that M. weeksiae and M. tenuis differed in sequence by 1.3%, M. weeksiae and M. platyphylla by 8.4%, and M. tenuis and M. platyphylla by 8.1%. Evaluation of standard marker genes indicated that sequences from the rbcL, RuBisCO spacer, and CO1 genes closely approximated the pair‐wise genetic distances observed between the plastomes of the three species of Membranoptera. A phylogenetic tree based on rbcL sequences showed that M. tenuis and M. weeksiae were sister taxa. Short rbcL sequences were obtained from type specimens of M. dimorpha, M. multiramosa, and M. edentata and confirmed their conspecificity with M. platyphylla. The data support the recognition of three species of Membranoptera occurring south of Alaska: M. platyphylla, M. tenuis, and M. weeksiae.     

A new species of Scutellaria (Lamiaceae) from California

Scutellaria holmgreniorum from Lassen Co., California, is described as new. A key to the species ofScutellaria in the Intermountain Region of western United States is provided.     

Population genetic structure of the South American species Hypochaeris lutea (Asteraceae)

The genus Hypochaeris has a recent evolutionary history caused by long‐distance dispersal in conjunction with adaptive radiation in the South American continent. Hypochaeris lutea is a perennial herb that grows mostly at altitudes of around 1000 m in cold swamps of the southern regions of Brazil. We investigated the amplified fragment length polymorphism (AFLP) in 270 individuals representing 11 Brazilian populations of H. lutea to elucidate the population genetic structure of this species. The frequencies of polymorphic loci and gene diversity ranged from 83.42% to 91.66% and from 0.26 to 0.34, respectively. Analysis of molecular variance revealed that most of the genetic variability was found within (76.67%) rather than among (23.3%) populations, agreeing with the pattern of genetic distribution within and among populations observed in other allogamous species of Hypochaeris. A Mantel test showed no correlation between genetic and geographic distances when all populations were considered. Simulations performed using a Bayesian approach consistently identified two clusters with different admixture proportions of individuals, as also revealed by a UPGMA dendrogram of populations. The pattern of genetic structure observed in H. lutea is consistent with a process of successive colonization events by long‐distance dispersal resembling the rapid and recent radiation that has been proposed to explain the origin of the South American species of Hypochaeris.