New genetic markers to identify European resistant abalone to vibriosis revealed by high-resolution melting analysis,a sensitive and fast approach

Increasing temperature of seawater is often associated with increased exposure incidence of disease in field and in aquaculture populations. Numerous episodic mass mortalities of the abalone Haliotis tuberculata have been observed along the northern Brittany coast of France caused by a complex interaction between the host, pathogen and environmental factors. Here, we evaluated the potential of high-resolution melting (HRM) analysis for mutation genotyping and development of genetic markers for resistance to vibriosis in the gastropod species H. tuberculata. Small amplicon assays were developed and revealed genetic polymorphism between surviving and susceptible abalone obtained after two successive infections of aquaculture families in controlled conditions. Together with specific COI haplotypes, we identified particular genotypes in nascent polypeptide-associated complex subunit alpha and ferritin genes linked to the susceptibility or resistance of abalone to vibriosis. Selection of genitors based on these genes may increase the proportion in offspring of resistant individuals of more than 76 %. Finally, HRM assays constitute a very efficient genotyping tool to validate the genetic markers on a representative number of individuals of wild populations and thus identify future resistant genitors for aquaculture or conservation purposes.     

Low genetic differentiation between isolated populations of the colonial ascidian Symplegma rubra Monniot, C. 1972

Pelagic larvae are highly important for maintaining the gene flow among populations of sessile marine invertebrates. Colonial ascidians consist, exclusively, of brooding species, with lecithotrophic larvae that have a limited dispersal. As a result, there is a marked differentiation among populations. In this work, we used allozyme electrophoresis to access the genetic variation in four populations of Symplegma rubra, a colonial ascidian frequently found in the intertidal zone of Southeastern Brazilian coast. High variability was found at three of the four sites sampled, the exception being Praia Grande in the State of Rio de Janeiro. At this site, there was a great preponderance of clones, which possibly reflected the enclosed nature of the location and its low water circulation that reduce the dispersal capabilities of these animals. S. rubra did not conform to expectations for random mating (Hardy–Weinberg equilibrium): there was a deficit of heterozygotes that was more related to the small population size than to inbreeding processes, since F _is analysis per locus revealed a deficiency of heterozygotes at only one locus—MDH*. The greatest variation in allele frequency was found for GPI-2*. Analyses of genetic variability revealed moderate differentiation among the populations (F _ST=0.051), which was unexpected for a species with a low dispersal capability. Rafting, a frequently underestimated means of dispersal, may be the main mode of gene flow in this species over large areas, since colonies of S. rubra are frequently seen growing on drift material and there is no evidence that the larva survive for a long time in the plankton.     

苯并[a]芘和9,10-二甲基蒽对皱纹盘鲍(Haliotis dis-cus hannai)早期发育的毒性研究

为探究海洋中多环芳烃(PAHs)对海洋生物的毒性作用,以皱纹盘鲍(Haliotis discus hannai)为受试生物,采用半静水式实验方式,探讨了2种多环芳烃苯并[a]芘和9,10-二甲基蒽对皱纹盘鲍早期发育的毒性效应。在不同浓度的苯并[a]芘和9,10-二甲基蒽作用下,观察皱纹盘鲍的卵子受精率、胚胎发育时间和幼体死亡率。结果表明,高浓度的多环芳烃处理组与对照组相比,卵子的受精率显著降低,胚胎发育时间和幼体死亡率显著增加,与处理浓度之间存在显著的剂量-效应关系(P<0.05)。3～12 d中,苯并[a]芘对皱纹盘鲍幼体的LC_(10)分别为11.6、8.18、7.67和7.66 mg·L~(-1),9,10-二甲基蒽对皱纹盘鲍幼体的LC10分别为14.91、14.11、12.82和9.64 mg·L~(-1)。苯并[a]芘毒性大于9,10-二甲基蒽。     

A genetic analysis of the pea crabs (Decapoda: Pinnotheridae) of New Zealand

Seven populations of the pea crabPinnotheres atrinicola Page were sampled from around the North Island of New Zealand from February to October 1987, and individuals were scored at 23 presumptive enzymatic loci. For a brachyuran crab,P. atrinicola revealed high levels of polymorphism and heterozygosity. Of the loci scored, phosphoglucose isomerase (Gpi) and phosphoglucomutase (Pgm) were distinguished by high variability ( =0.602 and 0.526, respectively). A clinal variation in electromorph frequency was evident at several loci, and atGpi in particular. Statistical analyses revealed that, despite relatively small genetic distance separation, a high degree of structuring was present between the geographic populations. The degree of population subdivision observed in this study is atypical of brachyuran crabs. It is suggested that the genetic differentiation observed between pea crab populations is maintained by life-history attributes and current movements which restrict gene flow between populations and, to some extent, by random genetic drift.Publication No. 41 from the Evolutionary Genetics Laboratory, University of Auckland     

Contrast in the importance of arrow squid as prey of male New Zealand sea lions and New Zealand fur seals at The Snares,subantarctic New Zealand

New Zealand sea lions (Phocarctos hookeri) are threatened by incidental bycatch in the trawl fishery for southern arrow squid (Nototodarus sloanii). An overlap between the fishery and foraging sea lions has previously been interpreted as one piece of evidence supporting resource competition for squid. However, there is currently no consensus about the importance of squid in the diet of New Zealand sea lions. Therefore, we investigated this importance independently of spatial and temporal differences in squid availability through a simultaneous study with sympatric New Zealand fur seals (Arctocephalus forsteri), a species known to target arrow squid. Diet sampling at The Snares (48°01′S 166°32′E), subantarctic New Zealand, in February 2012 coincided with peak annual catch in the nearby squid fishery. Diets were deduced by analyses of diagnostic prey remains from scats (faeces) and casts (regurgitations). The contribution of each prey species to the diet was quantified using the per cent index of relative importance (% IRI) that combined frequency of occurrence, mass and number of prey items. Arrow squid was a minor component in sea lion scats (2 % IRI), and none was found in their casts. In contrast, arrow squid was the major component in fur seal scats and casts (93 and 99 % IRI, respectively). This study found that New Zealand sea lions ate minimal squid at a time when squid was clearly available as evidenced by the diet of New Zealand fur seals; hence, there was no indication of resource competition between sea lions and the squid fishery.     

Tissue and sub-cellular distribution of Fe,Cu, Zn and 210Po in the abalone Haliotis rubra

The tissue and sub-cellular distribution of Fe, Cu, Zn and the naturally occurring radionuclide polonium-210 was determined in the gastropod mollusc Haliotis rubra collected from Western Port Bay, Australia, between March and July 1988. The highest concentrations of the metals, with the exception of Cu, were found in the digestive gland. Copper was more uniformly distributed, with tissues that are more vasculated having higher concentrations. Ultrastructural examination of the digestive gland, gill and kidney showed dense membrane-bound granules within the cytoplasm. Elemental analysis of the granules by electron probe x-ray microanalysis indicated that the granules in the digestive gland and gill contained high concentrations of iron, with small amounts of copper and zinc. In contrast, the metal-containing granules in the kidney were predominantly composed of iron, copper and phosphorus, with variable contributions of sodium potassium, and calcium. Homogenisation and fractionation of the digestive gland by differential centrifugation confirmed that approximately 80, 10, 90 and 50% of the total homogenate Fe, Cu, Zn and ²¹⁰Po, respectively, sedimented at 1200xg. In the haemolymph, all the elements studied were associated with the soluble high molecular weight component of the serum, not with the amoebocytes. ²¹⁰Po was present in the mucus-secreting hypobranchial glands at about half the concentration found in gill tissue.     

Recruitment to adult habitats following marine planktonic development in the fiddler crabs, Uca pugilator, U. pugnax, and U. minax

Three congeneric species of fiddler crabs, Uca pugilator (Bosc, 1802), U. pugnax (Smith, 1870), and U. minax (LeConte, 1855), co-occur in estuaries along the east coast of North America, from Cape Cod to northern Florida. Although U. minax adults are generally found at lower salinities than the other two species, the distributions of all three species overlap to some degree. The distribution of megalopae and juvenile fiddler crabs (from first crab stage to those with a carapace width of 3.0 mm) was examined at four sites along a salinity gradient (from 35.0±2.0 to 3.0±3.0; ) in the North Inlet Estuary, South Carolina, USA, in August 2002. A PCR-RFLP technique was developed to identify individuals from the genus Uca to species from first zoea through the early crab stages. An examination of the distribution of early life stages showed that U. minax reinvades low-salinity adult habitats at settlement, following planktonic larval development in the coastal ocean. Also, juveniles of U. pugilator were found to occupy Spartina alterniflora stands, where adult conspecifics rarely occur. Species frequencies were different for adults compared to early life stages in low-salinity areas of the marsh, where populations overlap. Settlement and survival dynamics of early life-history stages in wet and dry years likely determine the distribution of adult Uca spp. populations along salinity gradients in estuarine ecosystems.Communicated by J.P. Grassle, New Brunswick     

Application of a spatial meta-population model with stochastic parameters to the management of the invasive grass Nassella trichotoma in North Canterbury, New Zealand

Optimising the management of invasive plants requires the identification of the population size outcomes for alternative management strategies. Mathematical models can be useful tools for making such management strategy comparisons. In this paper we develop a generic landscape meta-population model and apply it to the weedy grass, Nassella trichotoma, an invasive species occupying approximately 800 land parcels, predominantly pastoral farms, in the Hurunui district, North Canterbury, New Zealand. Empirical evidence reveals that this meta-population is currently stable (at a median density of 6 plants ha⁻¹) under a community strategy requiring manual removal (termed ‘grubbing’) of plants annually from all land parcels. Reduction in population size requires an alternative management strategy. Field data, collected over a 12 year period, were used to provide stochastic parameter values for land parcel size, carrying capacity, rates of local population growth and grubbing.The model reveals that at steady state, the most significant contribution to population growth on a land parcel comes from within the land parcel itself; the expected annual per capita growth on an individual land parcel is 4 orders of magnitude greater than the expected annual contribution from plants arising from other land parcels. This result implies that many of the farms currently supporting N. trichotoma may pose little or no threat to, nor are threatened themselves by, other farms infested by the weed. However, the steady state distribution (of the weed's population density) was sensitive to the spread rate, revealing a need for data on this process. It was also sensitive to how any increase in the grubbing rate is distributed; increasing it via a uniform distribution U(0, 1) where all rates between 0 and 100% year⁻¹ are equally probable did not affect the steady state, whereas increasing the rates via the uniform distribution U(0.25, 0.75) resulted in fewer farms with high population densities. In general the model provides a basis for exploring the effects of a wide range of alternative grubbing strategies on population growth in N. trichotoma.     

Temporal stability of genetic population structure in the New Zealand snapper, <Emphasis Type="Italic">Pagrus auratus</Emphasis>, and relationship to coastal currents

Genetic diversity and population structure of snapper (Pagrus auratus, Bloch and Schneider), a coastal demersal sparid fish, were determined using six nuclear microsatellite loci and SSCP (single strand conformational polymorphism) analysis of themitochondrial (mt) DNA D-loop in samples collected across the range of the species in New Zealand. Microsatellite data showed similar results to allozyme data collected in the late 1970s that found differentiation between the north-east and southern populations. In addition, an isolated population of snapper in Tasman Bay was identified. The two data sets provide evidence for the temporal stability of the genetic population structure of snapper over 22 years, with differentiation over relatively small spatial scales separated by oceanographic boundaries rather than isolation by distance. In contrast to nuclear markers, mtDNA did not reveal any significant genetic heterogeneity among samples.     

14C标记的月桂酸盐在淡水勾虾（Hyalella azteca）中的生物富集、代谢及空间分布

对水的生物蓄积性监管评估通常是基于鱼类流水暴露试验得出生物浓缩因子（BCF）。这样的实验需要很多实验动物,而且费时费钱。最近,有人提出了一种替代性的有机中性化合物的测试方法,即用勾虾（Hyalella azteca）进行测试得到BCF值,该值与鱼类 BCF值有很强的相关性。本文研究了离子化合物月桂酸盐在H. azteca中的生物浓缩潜力。 采用流动和半静态的方法对勾虾H. azteca进行1-14C-月桂酸钠暴露实验。由于该化合物的生物降解速度快,需要采用半静态方法,即频繁更换介质,以确保介质浓度稳定。月桂酸盐也可被H. azteca快速代谢。在勾虾组织中测得的总放射性中有很大一部分是不能提取的,这表明矿化月桂酸盐富集在H. azteca的钙化外骨骼中。这在使用14C标记的碳酸盐的进一步研究中得到了证实。以游离脂肪酸计量,月桂酸盐的脂质标准化（5.0%）的勾虾 BCF为8.9。H. azteca的生物浓缩研究结果证实了先前在鱼类中观察到的试验项目的低生物富集潜力。然而,需要测试更多的具有不同性质的有机离子化合物,以评估鱼和勾虾的BCF数据之间是否存在一般相关性。     

Low levels of global genetic differentiation and population expansion in the deep-sea teleost <Emphasis Type="Italic">Hoplostethus atlanticus</Emphasis> revealed by mitochondrial DNA sequences

The orange roughy Hoplostethus atlanticus is a well-known commercial species with a global distribution. There is no consensus about levels of connectivity among populations despite a range of techniques having been applied. We used cytochrome c oxidase subunit I (COI) and cytochrome b sequences to study genetic connectivity at a global scale. Pairwise Φ_ST analyses revealed a lack of significant differentiation among samples from New Zealand, Australia, Namibia, and Chile. However, low but significant differentiation (Φ_ST = 0.02–0.13, P < 0.05) was found between two Northeast Atlantic sites and all the other sites with COI. AMOVA and the haplotype genealogy confirmed these results. The prevalent lack of genetic differentiation is probably due to active adult dispersal under the stepping-stone model. Demographic analyses suggested the occurrence of two expansion events during the Pleistocene period.     

Concordance of genetic and fin photo identification in the great white shark, Carcharodon carcharias, off Mossel Bay, South Africa

Visual identification of naturally acquired marks has been a popular if subjective method of animal identification and population estimation over the last 40 years. Molecular genetics has also independently developed objective individual marking techniques during the same period. Here, we assess the concordance of individual great white shark (Carcharodon carharias) dorsal fin recognition and identification, using seven microsatellite loci as the independent unbiased arbiter, over a period of 5 years. As a monitoring technique, fin photographs offer a very good individual identification key for white sharks over a relatively short period of time (5 years), matching with genetic data in about 85% of cases, whilst caution and a continuously updated database is required for animal recognition over a longer period.     

Population biology of the annual grass Triplasis purpurea in relation to distance from shore on Staten Island, New York

Four populations of the native annual grassTriplasis purpurea were surveyed on coastal beaches along the south shore of Staten Island, NY, to determine the potential of this species to colonize shoreline habitats mostly devoid of other vegetation. If the species can establish and maintain dense populations, it may have conservation value for urban beaches disturbed by human activities. For two populations, survivorship, growth, and reproduction were monitored at different distances from shore to determine the ability of this species to maintain viable populations. At three sites,T. purpurea occurred in >75% of all quadrats and the highest density was 1195 plants/m² at 74 m from shore in one recently disturbed site. Density generally increased with increasing distances from shore at low tide (from ca. 40–90 m). Plants showed the greatest growth and reproduction at close distances to shore (30–40 m); part of this effect was due to density in one population, but when density effects were removed statistically, there still remained a decline in growth and reproduction with increasing distance from shore. Improved vigor nearest to shore may be due to continual sand deposition. Survivorship showed a Type I pattern, with low mortality throughout the growing season. By colonizing newly-deposited and continually shifting sands,T. purpurea can contribute to the earliest stages of ecological succession along disturbed beaches in eastern North America and may be valuable to the development and management of urban coastal plant communities.     

Temporal limits to simulating the future spread pattern of invasive species: Buddleja davidii in Europe and New Zealand

The spread of invasive species is a major ecological and economic problem. Dynamic spread modelling is a potentially valuable tool to assist regional and central government authorities to monitor and control invasive species. To date a lack of suitable data has meant that most broad scale dispersal models have not been validated with independent datasets, and so their predictive ability and reliability has remained unscrutinised. A dynamic, stochastic dispersal model of the widely invasive plant Buddleja davidii was calibrated on European spread data and then used to project the temporal progression of B. davidii's distribution in New Zealand, starting from several different historical distributions. To assess the model's performance, we constructed an occupancy map based on the average number of simulation realisations that have a population present. The application of Receiver Operating Characteristic (ROC) curves to occupancy maps is introduced, but with specificity substituted by the proportion of available area used in a realisation. A derivative measure, the partial area under these curves when assessed through time (pAUC), is introduced and used to assess overall performance of the spread model. The model was able to attain a high level of model sensitivity, encompassing all of the known locations within the occupancy envelope. However, attempting to simulate the spread of this invasive species beyond a decade had very low model specificity. This is due to several factors, including the exponential process of spread (the further a population spreads the more sites exist from which it can spread stochastically), and the Markovian chain property of the stochastic system whereby differences between realisations compound through time. These features are seen in many reports of spread models, without being explicitly acknowledged. Our measure of pAUC through time allows a model's temporal performance and its specificity to be simultaneously assessed. While the rapid deterioration in model performance limits the utility of this type of modelling for forecasting long-term broad-scale strategic management of biological invasions, it does not necessarily limit its attractiveness for informing smaller scale and shorter term invasion management activities such as surveillance, containment and local eradication.     

Reproductive biology of the antipatharian black coral Antipathes fiordensis in Doubtful Sound, Fiordland, New Zealand

The reproductive biology of Antipathes fiordensis Grange, a species endemic to south-western New Zealand, was followed from April 1994 to May 1995. Ten colonies were individually tagged in Doubtful Sound and sampled on a monthly basis in order to determine their reproductive activity. The fecundity of each of the five tagged female colonies was determined by estimating the total number of polyps per colony from photographs of each colony and by planimetry, the proportion of gravid polyps per colony, and the mean number of oocytes per gravid polyp. In addition, 56 colonies were sampled in March 1995 to estimate the sex ratio, height at sexual maturity, and mean sizes of females and males. A. fiordensis was found to be a dioecious species which, in the absence of gonads in the polyps, has no obvious external morphological differences between the sexes. Broadcast spawning of gametes is the likely mode of reproduction. Gametogenesis began in November 1994 and was highly synchronous within and between colonies, with spawning occurring in March 1995. The sex ratio in adults was 1:1. Colonies reached sexual maturity between the heights of 70 and 105 cm which, based on existing estimates of growth rate, corresponds to a minimum age for sexual maturity of about 31 yr. The largest oocytes measured ranged from 100 to 140 μm in size. Female colonies produced between 1.3 and 16.9 million oocytes, with the larger colonies dominating the reproductive output of the population. Received: 23 June 1997 / Accepted: 1 August 1997     

The structure of subtidal algal and invertebrate assemblages at the Chatham Islands,New Zealand

We examined the distribution and abundance of organisms on subtidal rocky reefs at nine sites around the Chatham Islands, a remote group 780 km east of southern New Zealand. We sampled five depth strata ranging from 1 to<16 m to identify spatial patterns in the abundance of algae and invertebrates and to assess their variation within and among sites. This information is used to discuss hypotheses concerning community structure at this remote locality. Several patterns were apparent. The immediate subtidal was occupied by the southern bull kelp Durvillaea spp. A suite of 11 fucalean species were dominant to a depth of 10 m with an average abundance of 28 m^-2, while one species, Carpophyllum flexuosum, occurred mostly in deeper water. Only two laminarian species of algae were present at the islands. The indigenous Lessonia tholiformis was abundant at 2.5 to 15 m and was not found in deeper water, while the giant kelp Macrocystis pyrifera was abundant at two sites in 12 to 18 m. The commercially valuable abalone Haliotis iris was extremely abundant in shallow water, with an overall mean of 6 m^-2 at 5 m. The sea urchin Evechinus chloroticus was common, but reached high densities only in small (<25 m²) patches. The characteristic urchin-dominated zones reported in kelp beds world-wide were not seen. There was considerable site-to-site variation in the occurrence and abundance of individual species. Some differences between sites were associated with shelter from swell (e.g. M. pyrifera was found only in sheltered sites) and physical habitat (e.g. juvenile H. iris were found only beneath boulders inshore), but much of the variation could not be explained by physical or depth-related factors alone. We hypothesize that the differences in these kelp bed assemblages compared to mainland New Zealand are partially due to the high degree of endemism at the Chatham Islands. Local variation cannot be explained by herbivory, and is most likely the result of the various life-history characteristics of the major habitat-forming species, the large brown algae.     

No apparent genetic basis to demographic differences in scarid fishes across continental shelf of the Great Barrier Reef

 Two species of parrot fish, Scarus frenatus and Chlorurus sordidus, are known to exhibit demographic and life-history differences across the continental shelf of the northern Great Barrier Reef (GBR). DNA sequences from the mitochondrial (mt) control region were analysed to determine whether there were genetic differences between the populations from the mid- and outer-shelf reefs. Analysis of molecular variance (AMOVA) indicated high levels of gene exchange for both species at a local scale between reefs on mid- and outer continental shelf positions (20 km apart) and at a broader scale along the length of the GBR province (>1000 km apart). There was no evidence to suggest that local differences in scarid life-history characteristics on the northern GBR have a genetic basis. Rather it appears more likely that phenotypically plastic responses to prevailing social and environmental conditions explain differences in the life-history characteristics of both taxa. However, analysis of genetic variation and historical demography revealed striking differences between the two species. S. frenatus haplotypes differed from one another at relatively few nucleotide sites (mean = 3.30), and the pairwise mismatch distribution suggested this species has undergone a population expansion within the limit of the resolution of the marker. C. sordidus haplotypes, however, differed from one another at a number of sites (mean = 7.67). Mismatch distribution analysis suggested that the population size of this species has remained at equilibrium over time. These patterns could also reflect differences in the metapopulation sizes or generation times between taxa. Some of the implications for fisheries management are discussed. Received: 28 January 2000 / Accepted: 9 July 2000