Distribution of sea snakes in the Great Barrier Reef Marine Park: observations from 10 yrs of baited remote underwater video station (BRUVS) sampling

The distributions of three species of sea snake (olive sea snake: Aipysurus laevis, spine-bellied sea snake: Lapemis curtus, and ornate sea snake: Hydrophis ocellatus) were estimated over 14° of latitude within the Great Barrier Reef Marine Park (GBRMP) using data from baited remote underwater video stations (BRUVS). A total of 2,471 deployments of BRUVS were made in a range of locations, in sites open and closed to trawl fishing. Sightings of sea snakes were analysed alongside six spatial factors [depth, relative distance across (longitude) and along (latitude) the GBRMP, proximity to land, proximity to the nearest reef, and habitat complexity] to determine the factors that most strongly influenced the distribution and abundance of sea snakes. The results showed a strong latitudinal effect on the distribution of all three sea snake species, with the highest densities and diversities occurring in central and southern GBRMP locations, while the northern Great Barrier Reef was relatively depauperate in terms of both occurrence and diversity. Shallow inshore areas were identified as key habitats for A. laevis and L. curtus, whereas deeper offshore habitats were most important for H. ocellatus. No significant difference was found in the mean number of snakes sighted per hour between sites open and closed to trawling. There was a high degree of congruence in the distribution of sea snakes estimated from the BRUVS data and results from previous trawl and underwater visual surveys, demonstrating the utility of BRUVS to estimate distribution and relative abundance in these species of sea snake at broad spatial scales in a non-extractive manner.     

Distribution of two species of sea snakes, <Emphasis Type="Italic">Aipysurus laevis</Emphasis> and <Emphasis Type="Italic">Emydocephalus annulatus</Emphasis>, in the southern Great Barrier Reef: metapopulation dynamics,marine protected areas and conservation

Aipysurus laevis and Emydocephalus annulatus typically occur in spatially discrete populations, characteristic of metapopulations; however, little is known about the factors influencing the spatial and temporal stability of populations or whether specific conservation strategies, such as networks of marine protected areas, will ensure the persistence of species. Classification tree analyses of 35 years of distribution data (90 reefs, surveyed 1–11 times) in the southern Great Barrier Reef (GBR) revealed that longitude was a major factor determining the status of A. laevis on reefs (present = 38, absent = 38 and changed = 14). Reef exposure and reef area were also important; however, these factors did not specifically account for the population fluctuations and the recent local extinctions of A. laevis in this region. There were no relationships between the status of E. annulatus (present = 16, absent = 68 and changed = 6) and spatial or physical variables. Moreover, prior protection status of reefs did not account for the distribution of either species. Biotic factors, such as habitat and prey availability and the distribution of predators, which may account for the observed patterns of distribution, are discussed. The potential for inter-population exchange among sea snake populations is poorly understood, as is the degree of protection that will be afforded to sea snakes by the recently implemented network of No-take areas in the GBR. Data from this study provide a baseline for evaluating the responses of A. laevis and E. annulatus populations to changes in biotic factors and the degree of protection afforded on reefs within an ecosystem network of No-take marine protected areas in the southern GBR. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Tetranucleotide markers from the loggerhead sea turtle (Caretta caretta) and their cross-amplification in other marine turtle species

The loggerhead sea turtle (Caretta caretta) is a federally threatened species and listed as endangered by the World Conservation Union (IUCN). We describe primers and polymerase chain reaction (PCR) conditions to amplify 11 novel tetranucleotide microsatellite loci from the loggerhead sea turtle. We tested primers using samples from 22 females that nested at Melbourne Beach, Florida (USA). Primer pairs yielded an average of 11.2 alleles per locus (range of 4–24), an average observed heterozygosity of 0.83 (range 0.59–0.96), and an average polymorphic information content of 0.80 (range 0.62–0.94). We also demonstrate the utility of these primers, in addition to primers for 15 loci previously described, for amplifying microsatellite loci in four additional species representing the two extant marine turtle families: olive ridley (Lepidochelys olivacea), hawksbill (Eretmochelys imbricata), green turtle (Chelonia mydas), and leatherback (Dermochelys coriacea).     

Molecular evidence that the deadliest sea snake Enhydrina schistosa (Elapidae: Hydrophiinae) consists of two convergent species

We present a striking case of phenotypic convergence within the speciose and taxonomically unstable Hydrophis group of viviparous sea snakes. Enhydrina schistosa, the ‘beaked sea snake’, is abundant in coastal and inshore habitats throughout the Asian and Australian regions, where it is responsible for the large majority of recorded deaths and injuries from sea snake bites. Analyses of five independent mitochondrial and nuclear loci for populations spanning Australia, Indonesia and Sri Lanka indicate that this ‘species’ actually consists of two distinct lineages in Asia and Australia that are not closest relatives. As a result, Australian “E. schistosa” are elevated to species status and provisionally referred to Enhydrina zweifeli. Convergence in the characteristic ‘beaked’ morphology of these species is probably associated with the wide gape required to accommodate their spiny prey. Our findings have important implications for snake bite management in light of the medical importance of beaked sea snakes and the fact that the only sea snake anti-venom available is raised against Malaysian E. schistosa.     

Pomphorhynchus laevis (Müller) in the flounder, Platichthys flesus L., in the tidal River Thames: population structure, microhabitat utilization and reproductive status in the field and under conditions of controlled salinity

The infection of R. Thames flounders, Platichthys flesus L., at Fulham by the acanthocephalan parasite Pomphorhynchus laevix (Müller) is described in terms of parasite population structure, life-cycle organization, reproductive biology and host microhabitat utilization. The parasites demonstrated 100% prevalence in this tidal but essentially freshwater locality (intestinal intensity 34.47) and were overdispersed in the flounder population (variance/mean ratio = 10.39, k= 1.04). Overall about 11% of the worms occurred in peritoneal cavity sites, the remainder being firmly attached to the gut wall in the posterior region of the intestine and rectum. The P. laevis populations were reproductively active, demonstrating that the flounder acts as a significant final host in this locality. Viable larval stages were identified in Gammarus zaddachi Sexton, a predominantly estuarine amphipod, indicating that P. laevis is likely to be able to complete its life cycle at Fulham. Over half the female parasites examined were gravid and many of the non-gravid worms were inseminated but had not yet started egg production. The proportion of females gravid, the number of ovarian balls and the number of eggs in gravid females was seen to increase with worm size. Both the identity of the intermediate host and the reproductive status of P. laevis in the Thames flounders differ from the Pomphorhynchus/flounder system studied by Kennedy (1984) in the R. Avon, suggesting that the two P. laevis populations may belong at least to discrete subspecies or strains. Parasites in flounders maintained under laboratory conditions in fresh water, 50% sea water and 100% sea water showed similar population, microhabitat and reproductive characteristics to those observed in the field. This suggests that increased salinity has a negligible effect on established parasites in the short term, and therefore that salinity may not form a barrier to the survival and dispersal of the R. Thames parasite when the flounders return to sea.     

Isolation and characterization of microsatellite loci in the black rat snake (Elaphe obsoleta)

We obtained molecular markers useful for population level studies of the black rat snake (Elaphe obsoleta) by screening genomic DNA libraries enriched for dinucleotide, tetranucleotide, and pentanucleotide microsatellite repeats. Following sequencing of the positive clones, 11 pairs of primers were designed for polymorphic loci and their variability assessed in > 350 individuals from four populations in North America. The loci had between 9 and 40 alleles and observed heterozygosities ranged from 0.071 to 0.87. Some of these pairs of primers also successfully amplified DNA from two other snake species.     

Development of microsatellite markers for the European white elm (Ulmus laevis Pall.) and cross‐species amplification within the genus Ulmus

Ulmus laevis Pall. is a broad‐leaved deciduous tree with a central and eastern European distribution. We describe the development of six polymorphic microsatellite markers for this species. These markers were also tested for utility in U. americana, U. glabra, U. minor and U. pumila. One additional marker gave ambiguous results in U. laevis but amplified clearly in three other species. In U. laevis, the number of alleles observed per locus ranged from two to nine. Five loci showed polymorphism in at least one of the nontarget species tested.     

Isotopic signatures, foraging habitats and trophic relationships between fish and seasnakes on the coral reefs of New Caledonia

A predator’s species, sex and body size can influence the types of prey that it consumes, but why? Do such dietary divergences result from differences in foraging habitats, or reflect differential ability to locate, capture or ingest different types of prey? That question is difficult to answer if foraging occurs in places that preclude direct observation. In New Caledonia, amphibious sea kraits (Laticauda laticaudata and L. saintgironsi) mostly eat eels—but the species consumed differ between snake species and vary with snake body size and sex. Because the snakes capture eels within crevices on the sea floor, it is not possible to observe snake foraging on any quantitative basis. We used stable isotopes to investigate habitat-divergence and ontogenetic shifts in feeding habits of sympatric species of sea kraits. Similarities in δ15 N (~10.5‰) values suggest that the two snake species occupy similar trophic levels in the coral-reef foodweb. However, δ13C values differed among the eight eel species consumed by snakes, as well as between the two snake species, and were linked to habitat types. Specifically, δ13C differed between soft- vs. hard-substrate eel species, and consistently differed between the soft-bottom forager L. laticaudata (~ −14.7‰) and the hard-bottom forager L. saintgironsi (~ −12.5‰). Differences in isotopic signatures within and between the two sea krait species and their prey were consistent with the hypothesis of habitat-based dietary divergence. Isotopic composition varied with body size within each of the snake species and varied with body size within some eel species, reflecting ontogenetic shifts in feeding habits of both the sea kraits and their prey. Our results support the findings of previous studies based on snake stomach contents, indicating that further studies could usefully expand these isotopic analyses to a broader range of trophic levels, fish species and spatial scales.     

Antipredator tactics: a kin-selection benefit for defensive spines in coral catfish?

Morphological features that impair a predator's ability to consume a prey item may benefit individual prey; but what of features that prolong prey-handling but do not enhance prey survival? For example, a striped eel catfish Plotosus lineatus will be fatally envenomated if struck by its specialist predator, the greater sea snake Hydrophis major. Nonetheless, the catfish typically erects long, toxic pectoral and dorsal spines that increase prey-handling times for the snake by around eightfold. Because the catfish travel in swarms of closely-related individuals, the delay enforced by spines may enable the victim's swarm-mates to disperse before the snake is able to search for another meal. In keeping with that hypothesis, defensive spines tend to be longer in catfish from regions where the greater sea snake occurs, than from areas where the snake does not occur.     

Isolation and characterization of novel microsatellite markers from the Australian tiger snakes (Elapidae: Notechis) and amplification in the closely related genus Hoplocephalus

We provide details of seven microsatellite loci (out of 20 primer pairs designed) that exhibited the cleanest and strongest banding patterns for the Eastern tiger snake Notechis scutatus. These loci were used to screen 76 individuals from across the geographical range of N. scutatus and a further 14 individuals of the closely related and endangered broadheaded snake Hoplocephalus bungaroides. We observed large numbers of alleles per locus (14–52) and relatively high levels of heterozygosity (0.270–0.696) within the N. scutatus sample. These markers are also likely to be informative for work on H. bungaroides.     

Spatial and genetic structure of two sandhills oaks: Quercus laevis and Quercus margaretta (Fagaceae)

Quercus laevis Walt, (turkey oak) and Q. margaretta Ashe (scrubby post oak) are important scrub oaks in the sandhills forest communities of the Coastal Plain of the southeastern United States. We used allozyme loci and Ripley's L-statistics to examine clonal structure and spatial dispersion in these species. Q. laevis greater than 1.5 m in height were randomly dispersed on a scale of 0–40 m; smaller individuals (< 1.5 m) were slightly clustered on a scale of 0–12 m. Larger individuals separated by ≤1 m had 15% probability of being ramets of the same clone. Q. margaretta showed strong clustering on a scale of 0–20 m. Stems separated by ≤ 1 m had a 71% probability of being ramets of the same clone. Clonal offspring were strongly clustered about the presumed clonal parent: 50% fell within 0.50 m of this individual. Simulation modeling and direct comparison of adult and juvenile genotypes indicated that acorns are dispersed on a scale of tens of meters for both species, suggesting animal vectors such as squirrels or blue jays.     

Development of simple sequence repeats (SSRs) in olive tree (Olea europaea L.)

We report the development of microsatellites or simple sequence repeats (SSRs) in the olive tree (Olea europaea L.). Forty three positive clones obtained by the screening of a GA-enriched genomic library were sequenced and primers were designed for 13 microsatellite loci. Five primer pairs amplified polymorphic products of the expected size range. SSR polymorphism was explored in a set of 46 olive cultivars. A total of 26 alleles were detected for the five loci. Heterozygosity ranged from 0.46 to 0.71. Ninety one per cent of the cultivars had unique multilocus genotypes. Microsatellite segregation was studied in a complex population from a cross between the commercial cultivars ’Leccino’ and ’Dolce Agogia’. Received: 3 February 2000 / Accepted: 21 March 2000     

Isolation and characterization of polymorphic microsatellite loci in the olive flounder (Paralichthys olivaceus)

We described the isolation and characterization of 27 new microsatellite loci from olive flounder, Paralichthys olivaceus. All loci were found to be polymorphic, and had between five and 22 alleles with observed heterozygosity ranging from 0.161 to 1.0 in 31 individuals examined. These micorsatellite makers are likely to be useful for studies of genome mapping, mating systems and population genetics in this species.     

Large‐scale connectivity,cryptic population structure,and relatedness in Eastern Pacific Olive ridley sea turtles (Lepidochelys olivacea)

Endangered species are grouped into genetically discrete populations to direct conservation efforts. Mitochondrial control region (mtCR) haplotypes are used to elucidate deep divergences between populations, as compared to nuclear microsatellites that can detect recent structuring. When prior populations are unknown, it is useful to subject microsatellite data to clustering and/or ordination population inference. Olive ridley sea turtles (Lepidochelys olivacea) are the most abundant sea turtle, yet few studies have characterized olive ridley population structure. Recently, clustering results of olive ridleys in the Eastern Tropical Pacific Ocean suggested weak structuring (F_ST = 0.02) between Mexico and Central America. We analyzed mtCR haplotypes, new microsatellite genotypes from Costa Rica, and preexisting microsatellite genotypes from olive ridleys across the Eastern Tropical Pacific, to further explore population structuring in this region. We subjected inferred populations to multiple analyses to explore the mechanisms behind their structuring. We found 10 mtCR haplotypes from 60 turtles nesting at three sites in Costa Rica, but did not detect divergence between Costa Rican sites, or between Central America and Mexico. In Costa Rica, clustering suggested one population with no structuring, but ordination suggested four cryptic clusters with moderate structuring (F_ST = 0.08, p < .001). Across the Eastern Tropical Pacific, ordination suggested nine cryptic clusters with moderate structuring (F_ST = 0.103, p < .001) that largely corresponded to Mexican and Central American populations. All ordination clusters displayed significant internal relatedness relative to global relatedness (p < .001) and contained numerous sibling pairs. This suggests that broadly dispersed family lineages have proliferated in Eastern Tropical Pacific olive ridleys and corroborates previous work showing basin‐wide connectivity and shallow population structure in this region. The existence of broadly dispersed kin in Eastern Tropical Pacific olive ridleys has implications for management of olive ridleys in this region, and adds to our understanding of sea turtle ecology and life history, particularly in light of the natal‐homing paradigm.     

Immunolocalization of Na/K–ATPase and Na/K/2Cl cotransporter in the tubular epithelia of sea snake salt glands

The sublingual salt gland is the primary site of salt excretion in sea snakes; however, little is known about the mechanisms mediating ion excretion. Na⁺/K⁺–ATPase (NKA) and Na⁺/K⁺/2Cl⁻ cotransporter (NKCC) are two proteins known to regulate membrane potential and drive salt secretion in most vertebrate secretory cells. We hypothesized that NKA and NKCC would localize to the basolateral membranes of the principal cells comprising the tubular epithelia of sea snake salt glands. Although there is evidence of NKA activity in salt glands from several species of sea snake, the localization of NKA and NKCC and other potential ion transporters remains unstudied. Using histology and immunohistochemistry, we localized NKA and NKCC in salt glands from three species of laticaudine sea snake: Laticauda semifasciata, L. laticaudata, and L. colubrina. Antibody specificity was confirmed using Western blots. The compound tubular glands of all three species were found to be composed of serous secretory epithelia, and NKA and NKCC were abundant in the basolateral membranes. These results are consistent with the morphology of secretory epithelia found in the rectal salt glands of marine elasmobranchs, the nasal glands of marine birds and the gills of teleost fishes, suggesting a similar function in regulating ion secretion.     

Cannibalism or congeneric predation? The African clawed frog,Xenopus laevis (Daudin), preferentially predates on larvae of Cape platannas,Xenopus gilli Rose & Hewitt

Predators are not limited to prey from other species as they can cannibalise vulnerable individuals within their own population. The African clawed frog, Xenopus laevis (Daudin), is a predator with a broad diet, known to consume multiple prey species, including congeners and conspecifics. African clawed frogs occur in sympatry with the Endangered Cape platanna, Xenopus gilli Rose & Hewitt, which are under threat through competition and predation from X. laevis. We investigated the threat of X. laevis predation on X. gilli using choice and no‐choice experiments to evaluate the relative vulnerability of X. laevis and X. gilli larvae. Results showed that large X. gilli larvae had a significantly higher vulnerability to X. laevis predation compared to small X. gilli larvae. However, the same discrimination was not discerned when offered large and small X. laevis larvae, or mixed larvae of the same size. We report ontogenic shifts in behaviour of X. gilli larvae that may be a factor in contributing to the vulnerability of large X. gilli larvae to adult X. leavis predation. Congeneric predation likely has negative implications for the population structure of the Endangered X. gilli. Our study underlines the call for the removal of X. laevis to conserve populations of X. gilli.     

Transferability of olive microsatellite loci across the genus <Emphasis Type="Italic">Olea</Emphasis>

The transferability of microsatellite markers developed for olive cultivars (Olea europaea L.) has been tested and confirmed in the Olea complex. Thirty two genotypes, belonging to different taxa of the genus Olea, have been analyzed with four olive SSRs. Positive amplifications at all loci were obtained in 13 taxa (at least one accession per species). Sixty seven different alleles have been detected at the four loci analyzed. Polymorphic products have been observed at the inter- and intra-species level. Some SSR loci have shown multiple amplification products in some species. The high number of unique alleles has allowed the unambiguous discrimination of most accessions. Similarity coefficients and relationships among the Olea taxa have been calculated based on SSR amplification results. The reliability of SSRs as markers for intra-species variability evaluation has been confirmed while their use to explore relationships at the inter-species level is discussed, being dependent on the locus analyzed.Communicated by H.F. Linskens