Genetic structure of<Emphasis Type="Italic"> Linckia laevigata</Emphasis> and<Emphasis Type="Italic"> Tridacna crocea</Emphasis> populations in the Palawan shelf and shoal reefs

Allozyme variation of 10 populations of Linckia laevigata at 8 polymorphic loci and 13 populations of Tridacna crocea at 6 polymorphic loci were analyzed to compare genetic variability and genetic affinities among reefs in Palawan, Philippines. Two to five populations were sampled from each of four regions: the shelf reefs in (1) northern Palawan and (2) southern Palawan and the offshore reefs in (3) the Kalayaan island group (KIG) in the South China Sea and (4) the Tubbataha shoals in the Sulu Sea. Heterozygosity was highest in populations of L. laevigata from the south shelf of Palawan and populations of T. crocea from the Tubbataha shoals of the Sulu Sea. The lowest heterozygosity estimates were from the reefs of the KIG in the South China Sea, for both species. Overall F _ST values for both species were significant, with an estimated average number of effective migrants per generation (N _EM) of 4.85 (~5 individuals) for L. laevigata and 3.54 (~4 individuals) for T. crocea. Within-region comparisons showed N_EM ranging from 6.29 to 92.34 for L. laevigata and from 3.40 to 6.30 for T. crocea. The higher gene flow among L. laevigata populations relative to T. crocea is consistent with the greater dispersal potential of the former species. Finer scale genetic structuring was evident in T. crocea populations. For both species, the Tubbataha reefs in the Sulu Sea have higher genetic affinity with the populations from the southern shelf of Palawan, while the reefs in the KIG had higher affinity with the northern Palawan shelf reefs. The north and south shelf populations have the least genetic affinity. Genetic patchiness among reefs within regions suggests the importance of small-scale physical factors that affect recruitment success in structuring populations in small island and shoal reef systems in Palawan.     

Physiological adaptations of the invasive cordgrass<Emphasis Type="Italic"> Spartina anglica</Emphasis> to reducing sediments: rhizome metabolic gas fluxes and enhanced O<Subscript>2</Subscript> and H<Subscript>2</Subscript>S transport

Cordgrasses of the genus Spartina form dense monospecific stands worldwide, profoundly influencing the ecology of estuaries. One species, Spartina anglica, originated by allopolyploidy in the 1800s and has been particularly prolific as an invasive species worldwide. S. anglica tolerates low-lying estuarine mudflats that its progenitor species and other coastal halophytes cannot. However, very little is known of the physiology of S. anglica. In the present study, an automated flow-through respirometry system was used to quantify metabolic gas fluxes (O₂, H₂S, CO₂, and NH₃) of S. anglica rhizomes. Enhanced physiological mechanisms to transport O₂ and H₂S in both directions between the rhizosphere and the atmosphere were exhibited by S. anglica, but not by the native North American species S. alterniflora. These results suggest that tolerance of anoxia and H₂S may assist S. anglica in colonizing extensively flooded environments. Enhanced sediment oxygenation by S. anglica may be potentially useful for phytoremediation of contaminated sediments, since microbial degradation of organic pollutants is often limited by O₂ availability.     

Patterns of gamete incompatibility between the blue mussels<Emphasis Type="Italic"> Mytilus edulis</Emphasis> and<Emphasis Type="Italic"> M</Emphasis>.<Emphasis Type="Italic"> trossulus</Emphasis>

Previous research on gametic incompatibility in marine invertebrates suggests that for highly dispersive marine invertebrate species, barriers to fertilization among closely related taxa are often incomplete and sometimes asymmetric. The nature of these barriers can dramatically affect the patterns of gene flow and genetic differentiation between species, and thus speciation. Blue mussels, in the genus Mytilus, are genetically distinct in allopatry yet hybrids are present wherever any two species within the group co-occur. The present study sampled M. edulis (L.) and M. trossulus (Gould) in May and June 2001 from the East Bay section of Cobscook Bay, Maine, USA (latitude 44°56′30″N; longitude 67°07′50″W), where the two species are sympatric. Gamete incompatibility was investigated in a series of laboratory fertilizations carried out in July 2001. The proportion of fertilized eggs typically exceeded 80% at sperm concentrations of 10³–10⁴ ml^?1 among intraspecific matings (n=18), but was <30% even at sperm concentrations in excess of 10⁵–10⁶ ml^?1 for interspecific matings (n=13). Further analysis indicated that approximately 100- to 700-fold higher sperm concentrations were required to achieve 20% fertilization in interspecific matings relative to intraspecific matings, indicating strong barriers to interspecific fertilization. The proportion of fertilized eggs did not follow this general pattern in all matings, however. The eggs from two (out of five) M. edulis females were almost as receptive to M. trossulus sperm as they were to M. edulis sperm. In contrast, the eggs from all M. trossulus females (n=3) were unreceptive to M. edulis sperm, suggesting that fertilization barriers between these species may be asymmetric. Given the experimental design employed in this study, the results are also consistent with a strong maternal or egg effect on the level of interspecific gamete compatibility in M. edulis.     

Developmental and genetic evidence for the existence of three morphologically cryptic species of<Emphasis Type="Italic"> Crepidula</Emphasis> in northern Chile

The taxonomy of Crepidula has been based principally on shell morphology, but shell characteristics in this genus may be strongly influenced by the substrates they inhabit and are thus of limited use in classification. Four species in this genus are currently recognized from the Chilean coast, of which two, Crepidula dilatata and C. fecunda, are sympatric. These species are morphologically cryptic and differ only in their larval development: C. dilatata has indirect development with a planktonic phase, and C. fecunda direct development within egg capsules. The other two species, C. coquimbensis and C. philippiana, are species that occur within gastropod shells inhabited by hermit crabs and differ from the other species in internal morphology of the shell, developmental features, and geographic distribution. Analyses of nine morphological characters, eight developmental characters, and 26 enzymatic loci were carried out on four Crepidula populations in the Coquimbo region of northern Chile. Four populations of Crepidula were identified based on morphological, developmental, and protein electrophoresis features. Our results suggest the presence of C. coquimbensis plus three morphologically cryptic species including C. dilatata, C. fecunda, and a new previously unrecognized cryptic species. All four populations showed differences in larval development pattern. C. fecunda have indirect development, with free-swimming larvae. The remaining three species have intracapsular metamorphosis, but different modes of larval nutrition. C. dilatata have nurse eggs that are consumed by mechanical destruction; C. coquimbensis engulf nutritional embryos; and the new species eat nutritive embryos by rotation. Protein electrophoresis data support the specific condition of each group with different larval development and endorse the use of developmental differences in taxonomy of Crepidula. Genetic distance shows that C. dilatata and C. fecunda are closely related, and that C. coquimbensis is closer to them than to the new cryptic species.     

Differences in stress tolerance and brood size between a non-indigenous and an indigenous gammarid in the northern Baltic Sea

Differences in stress tolerance and reproductive traits may drive the competitive hierarchy between non-indigenous and indigenous species and turn the former ones into successful invaders. In the northern Baltic Sea, the non-indigenous Gammarus tigrinus is a recent invader of littoral ecosystems and now occupies comparable ecological niches as the indigenous G. zaddachi. In laboratory experiments on specimens collected between June and August 2009 around Tvärminne in southern Finland (59°50′N/23°15′E), the tolerances towards heat stress and hypoxia were determined for the two species using lethal time, LT₅₀, as response variable. The brood size of the two species was also studied and some observations were made on maturation of juveniles. Gammarus tigrinus was more resistant to hypoxia and survived at higher temperatures than G. zaddachi. Brood size was also greater in G. tigrinus than in G. zaddachi and G. tigrinus matured at a smaller size and earlier than G. zaddachi. Hence, there are clear competitive advantages for the non-indigenous G. tigrinus compared to the indigenous G. zaddachi, and these may be further strengthened through ongoing environmental changes related to increased eutrophication and a warming climate in the Baltic Sea region.     

Evolution of coral reef fish<Emphasis Type="Italic"> Thalassoma</Emphasis> spp. (Labridae). 1. Molecular phylogeny and biogeography

Wrasses in the genus Thalassoma comprise 27 recognized species that occur predominantly on coral reefs and subtropical rocky reefs worldwide. The phylogenetic relationships for 26 species were examined based on two mitochondrial genes (cytochrome b and 16S rRNA) and one nuclear intron (the first intron of the ribosomal protein S7). Two closely related species, the bird-wrasses (Gomphosus varius Lacepède, 1801 and G. caerulaeus Lacepède, 1801), were also included in the analysis. These species grouped within the genus Thalassoma. Thalassoma newtoni (Osório, 1891) from Sao Tome, which is generally synonymized with the Atlantic/Mediterranean Thalassoma pavo (Linnaeus, 1758) appears to be a valid species. Using a molecular clock, the genus was estimated to have originally diverged 8–13 million years ago, with Thalassoma ballieui (Vaillant and Sauvage, 1875) from Hawaii and Thalassoma septemfasciata Scott, 1959 from Western Australia as the ancestral species. Approximately 5–10 million years ago, a sudden burst of speciation resulted in seven clades, which were resolved with the sequence data. The terminal Tethyan event and the closing of the Isthmus of Panama were probably the major historical factors shaping the evolution of species in the genus Thalassoma. These data on the spatio-temporal pattern of speciation in the Indo-Pacific indicate that peripheral species have been generated at various times throughout the history of the genus, and that none of the widespread species are relatively young. Thus, there is no clear support for centrifugal (youngest at the periphery) versus centripetal (oldest at the periphery) modes of generation of species, two theories which have been used to account for geographic gradients in species diversity.     

A population comparison of the strength and persistence of innate colour preference and learning speed in the bumblebee <Emphasis Type="Italic">Bombus terrestris</Emphasis>

Studies of innate colour preference and learning ability have focused on differences at the species level, rather than variation among populations of a single species. Initial strength and persistence of colour preferences are likely to affect colour choices of naïve flower visitors. We therefore study the influence of both the strength and persistence of innate colour preference (for blue) on an operant learning task (associating food reward with yellow flowers) in two populations of the bumblebee Bombus terrestris. We found that both strength and persistence of blue preference differed significantly between populations: B. terrestris dalmatinus had a weaker and less persistent blue preference than B. terrestris audax. These differences in preference also influenced learning performance. Considering only landing behaviours, one-trial learning occurred in the majority (73%) of bees, and was achieved sooner in B. terrestris dalmatinus because of its weaker blue preference. However, compared to landing behaviours the relative frequency of approach flights to rewarding and unrewarding flower types changed more slowly with task experience in both populations. When considering both approaches and landings, the rate of learning, following the first rewarded learning trial, was faster in B. terrestris audax than B. terrestris dalmatinus. However, the net effects of population differences in blue preference and learning dynamics result in similar final levels of task performance. Our results provide new evidence of behavioural differences among isolated populations within a single species, and raise intriguing questions about the ecological significance and adaptive nature of colour preference.     

Intraspecific variation of photosynthesis,respiration and photoprotective carotenoids in<Emphasis Type="Italic"> Gracilaria birdiae</Emphasis> (Gracilariales: Rhodophyta)

In most rhodophytes, the quenchers zeaxanthin or lutein seem to be used for carotenoid photoprotection next to β-carotene. However, our study shows that the tropical intertidal alga Gracilaria birdiae possesses high concentrations of antheraxanthin, as well as lower concentrations of both zeaxanthin and violaxanthin. In higher plants and some algal groups, these three oxygenated carotenoids are known to protect photosystem II in a xanthophyll cycle. We were able to manipulate the carotenoid concentrations in G. birdiae by varying irradiance and temperature in laboratory experiments. Higher concentrations of violaxanthin were found in dark incubations and higher concentrations of zeaxanthin in light incubations, which is an indication that a xanthophyll cycle is active in G. birdiae. Our results also suggest a biosynthesis pathway from lycopene to β,β-carotene and β-cryptoxanthin in the light and synthesis of new lycopene in darkness. This would imply that the production of functional carotenoids is regulated by their need in photoprotection. Despite cultivation in the laboratory under the same conditions for at least 6 years, there were differences in photosynthetic properties and carotenoid dynamics between algae obtained from a population close to the Equator and another population close to the Tropic of Capricorn. These differences most probably have their origin in genetic differentiation mediated by ambient irradiance and temperature regimes. Four phycoerythrin-deficient mutant strains of G. birdiae were studied as well. Photosynthetic properties and carotenoid dynamics were basically the same in the mutants and the wild strains, but two green strains had remarkably low dark respiratory rates. This is an indication of low production of metabolites, which may be a competitive disadvantage.     

Variation in fertilisation abilities between hemiclonal hybrid and sexual parental males of sympatric water frogs (<Emphasis Type="Italic">Rana lessonae</Emphasis>,<Emphasis Type="Italic"> R. esculenta</Emphasis>,<Emphasis Type="Italic"> R. ridibunda</Emphasis>)

In many species, males and females mate with multiple partners, which gives rise to sperm competition and multiple paternity. The experiments on water frogs presented here demonstrate that such sperm competition can affect the structure and dynamics of mixed-species communities. The hybrid frog Rana esculenta (LR) mates with one of its parental species, usually R. lessonae (LL), although in some areas R. ridibunda (RR), to regain the premeiotically eliminated parental genome ("hybridogenesis"). Mixed LL/LR-populations are stable although hybrid numbers should continuously increase at the expense of parental animals, because of differences in female fecundity and other factors. This would finally lead to the extinction of the sexual host, followed by that of the sexual parasite, unless the reproductive superiority of R. esculenta is reduced by other factors, such as lower hybrid male fertility. Eggs from LL- and LR-females were fertilised in vitro by single- and multi-male sperm suspensions of LL-, LR- and RR-males. In all experiments, the proportion of offspring sired by R. esculenta sperm was significantly lower than that sired by R. lessonae or R. ridibunda sperm. Gonad mass, sperm morphology, sperm swimming velocity, and sperm survival did not explain these differences in fertilisation success; nor did gamete recognition and compatibility. Sperm density was the only trait that paralleled fertilisation success, but it offers no explanation either, because densities were equalised for the in-vitro fertilisations. In natural LL/LR populations, the significantly smaller amount, poorer competitive ability and lower long-term survival of R. esculenta compared to R. lessonae sperm will reduce the initial reproductive superiority of hybrids and contribute to the stabilisation of mixed water-frog populations. Differences in fertilisation ability are also likely to be relevant for the structure and dynamics of several other systems with encounters between eggs and sperm from different genotypes, ecotypes, ploidy levels and/or species.     

Population structure of two northern hemisphere polychaetes,<Emphasis Type="Italic"> Neanthes virens</Emphasis> and<Emphasis Type="Italic"> Hediste diversicolor</Emphasis> (Nereididae), with different life-history traits

The present study compared population genetic structure of two northern hemisphere polychaetes, Neanthes virens (Sars, 1835) and Hediste diversicolor (O.F. Müller, 1776), with different life-history traits. N. virens has a pelagic larval phase that may last as long as 9 days, while H. diversicolor broods its offspring. It was predicted that N. virens populations, especially over relatively short geographic distances, would be more genetically similar than H. diversicolor populations. Sequence data were obtained for two mitochondrial genes, cytochrome c oxidase I (COI) and cytochrome b (cyt b). The COI gene showed no variation in 16 N. virens individuals from 12 locations, and it was not examined in H. diversicolor. The nucleotide sequence of the cyt b gene was examined in 93 N. virens individuals collected in 1992, 1999, and 2000 from 12 localities throughout its range (North America, Europe, Japan) and from 33 H. diversicolor individuals collected in 1999 and 2001 from one locality in eastern Canada and two localities in France (Roscoff and Marseille). In N. virens, one cyt b haplotype was shared by 89% (83 out of 93) and only two other haplotypes were found. Significant population structure was observed in H. diversicolor. In the 33 sequences compared, 18.9% of sites were polymorphic and 12 haplotypes were identified. There were two haplotypes in the Canadian population, three in the Marseille samples, and seven in the Roscoff samples, and no haplotype was found in common between any two sites. These results show that widely separated H. diversicolor populations are strongly differentiated at the cyt b locus. The most plausible interpretation for the low variation at the COI and cyt b loci in N. virens, and for the lack of variation at allozyme loci (based on an initial survey of a smaller subset of populations), is that this species has experienced a drastic bottleneck and that a few remaining lineages recolonized its entire range.     

Evaluation of microelement contents in <Emphasis Type="Italic">Clethra barbinervis</Emphasis> as food for human and animals in contrasting geological areas

The young leaves of Clethra barbinervis Sieb. et Zucc, which is a deciduous tree species found in secondary forests widely in Japan, are used in spring as a local traditional food by local populations, and the bark of this plant is also preferred by sika deer, Cervus nippon. However, C. barbinervis has been known to accumulate heavy metals in its leaves. Then, we aimed to clarify the characteristics of microelement contents in C. barbinervis and to discuss the value of this species as food for humans and animals through the analysis of seasonal changes and distribution in various organs of C. barbinervis growing under two different geological conditions. We found that C. barbinervis is an accumulating and tolerant plant for Ni, Co and Mn. It accumulates Ni from serpentine soil containing Ni at high concentration, and Co and Mn from acidic soils based on crystalline schist. The seasonal variation in element concentrations in leaves indicates that the young leaves contain Cu at high concentration and that eating them in spring season may be advantageous to humans, due to the associated increase in Cu intake. The high concentrations of Cu and Zn in the bark of C. barbinervis might explain why deer prefer to eat the bark of this species.     

Four genes,morphology and ecology: distinguishing a new species of <Emphasis Type="Italic">Acesta</Emphasis> (Mollusca; Bivalvia) from the Gulf of Mexico

There is currently an active scientific debate about the most correct and efficient way to identify species. To date, few studies in the marine realm have combined the available taxonomic methods. In this study we have used morphology, ecology and molecular analyses to identify a new species within the bivalve genus Acesta. All four genes studied (12S, 16S, Cytb, COI) suggested that a common cold-seep species in the Gulf of Mexico, A. bullisi, should be divided into two distinct species. This conclusion was supported by morphological traits and by observations of ecological distribution. A. oophaga Järnegren, Schander and Young n. sp. is described here, and A. bullisi Vokes (Tulane Stud Geol 2:75–92, 1963) is re-described. This study shows that DNA barcoding in combination with traditional morphological and ecological analyses may be an important tools to identify hidden biodiversity among deep-water organisms such as bivalves.     

Phytotoxic effects of phenolic acids from <Emphasis Type="Italic">Merostachys riedeliana,</Emphasis> a native and overabundant Brazilian bamboo

Merostachys riedeliana Rupr. is a native and overabundant bamboo species in the Brazilian Atlantic Forest. Moderate to strong allelopathic activity may be one mechanism that explains this super-dominance and the changes in structure and composition of forest areas occupied by bamboo. This study evaluated the phytotoxic effect of M. riedeliana extracts and fractions and identified their putative allelochemicals. We investigated the presence of allelochemicals in soil collected from stands occupied by M. riedeliana. Furthermore, we evaluated the putative effect of tree allelochemicals, individually and combined, on germination and growth. The aqueous extract of leaves and its ethyl acetate fraction presented the highest inhibitory effects on seed germination and seedling growth. The effect of the extracts and fractions on the target species was species-specific. Neither the individual nor the combined phenolic acids significantly inhibited seed germination; however, a pronounced growth inhibition was observed in M. bimucronata seedlings treated with vanillic acid and in E. verna and M. bimucronata seedlings treated with combined phenolic acids. Isovitexin, vitexin, isoorientin, orientin, and their O-glycoside derivatives, the lactonic dimer of the p-hydroxybenzoic acid and 3,4-methylenedioxymandelic acid were identified in the aqueous extracts and ethyl acetate fraction by Liquid Chromatography-Diode Array Dectector/Electrospray Ionization/Mass Spectrometry (LC-DAD/ESI–MS/MS). The Gas Chromatography-Mass Spectrometer (GC–MS) profile of the same extract and fraction showed the presence of benzoic, benzeneacetic, salicylic, p-hydroxybenzoic, p-hydroxyphenylacetic, vanillic, p-coumaric, protocatechuic, syringic, gallic, m-coumaric vanillylmandelic, 4-methylmandelic, 3,4-methylenedioxymandelic and trans-ferulic acids. The p-benzoic acid and the apigenin 6-C-glucoside (isovitexin) were identified in the soil extract collected from under bamboo-growing areas. Even though laboratory bioassays are not completely predictive of the allelopathic effects that occur in nature, the results of this study provide preliminary evidence of allelopathy as a possible species-specific inhibition mechanism of native species that explain the impoverishment of floristic richness and the functional groups in areas where M. riedeliana is overabundant.     

Differences in the species- and size-composition of fish assemblages in three distinct seagrass habitats with differing plant and meadow structure

Fish faunas were sampled seasonally using a large and a small beam trawl in three seagrass habitats comprising predominantly Amphibolis griffithii or Posidonia sinuosa or Posidonia coriacea, which differ in seagrass and meadow structure. Amphibolis griffithii and P. sinuosa both produce a relatively dense leaf canopy, but the former exhibits a distinct architecture, with the leaf canopy overlying relatively open spaces surrounding woody stems, compared to the uniformly dense blade-like leaves of P. sinuosa which emerge directly from the sediment. In comparison, P. coriacea provides a landscape of patchy seagrass amongst areas of bare sand. Since the latter seagrass habitat contains large areas of sand, fish were also sampled in adjacent unvegetated areas. The number of species and density of fish were greater (P<0.05) in P. sinuosa than in either A. griffithii or P. coriacea. The mean number of species caught using the large trawl ranged from 16 to 24 in the first of these habitats compared to 14–21 and 9–15 in the last two habitats, respectively, and the mean densities ranged between 78 and 291 fish 1000 m^?2 in P. sinuosa compared to 31–59 fish 1000 m^?2 in A. griffithii and 31–59 fish 1000 m^?2 in P. coriacea. The biomass of fish was greater (P<0.05) in both P. sinuosa and A. griffithii than in P. coriacea (4.2–5.3 kg and 3.3–6.2 kg versus 0.7–1.9 kg 1000 m^?2, respectively). Furthermore, the size-structure of fish differed among these habitats, where the median weight of fish was 72.1 g in A. griffithii, compared to 7.5 g and 19.8 g in P. sinuosa and P. coriacea, respectively. Ordination and ANOSIM demonstrated that the species-composition differed markedly among the three seagrass habitats (P<0.001), suggesting that fish species display a distinct preference for particular seagrasses characterised by different architecture. Differences in species-composition among the seagrass habitats partly reflected the size-composition of fish in each habitat. The open space below the canopy of A. griffthii is likely to allow larger fish to occupy this habitat, whereas only small fish would be able to penetrate the dense foliage of P. sinuosa. Differences in species- and size-composition of fish among these habitats may be the result of settlement-sized larvae discriminating between particular seagrass and meadow structures, or fish being subject to different levels of predation and/or accessibility to food or space. The species-composition in P. coriacea was highly dispersed and did not differ from that of unvegetated areas. While several species were associated with both P. coriacea and bare-sand habitats, some species did display a high affinity with the seagrass P. coriacea. This may reflect an association with the sparse and narrower leaves of this seagrass or with the patchy occurrence of the seagrass Heterozostera tasmanica, which commonly occurs as an understorey in this habitat.     

Effect of ultraviolet irradiation and chlorination on ampicillin-resistant <Emphasis Type="Italic">Escherichia coli</Emphasis> and its ampicillin resistance gene

Antibiotic resistance is a serious public health risk that may spread via potable and reclaimed water. Effective disinfection is important for inactivation of antibiotic-resistant bacteria and disruption of antibiotic resistance genes. Ampicillin is a widely prescribed antibiotic but its effectiveness is increasingly undermined by resistance. In this study, changes in ampicillin resistance for Escherichia coli (E. coli) CGMCC 1.1595 were analyzed after exposure to different doses of ultraviolet (UV) or chlorine, and damage incurred by the plasmid encoding ampicillin resistance gene bla _TEM-1 was assessed. We reported a greater stability in ampicillinresistant E. coli CGMCC 1.1595 after UV irradiation or chlorination when compared with previously published data for other E. coli strains. UV irradiation and chlorination led to a shift in the mortality frequency distributions of ampicillin-resistant E. coli when subsequently exposed to ampicillin. The ampicillin hemiinhibitory concentration (IC₅₀) without disinfection was 3800 mg·L^–1, and an increment was observed after UV irradiation or chlorination. The IC₅₀ of ampicillin-resistant E. coli was 1.5-fold higher at a UV dose of 40 mJ·cm^–2, and was 1.4-fold higher when exposed to 2.0 mg·L^–1 chlorine. These results indicate that UV irradiation and chlorination can potentially increase the risk of selection for E. coli strains with high ampicillin resistance. There was no evident damage to bla _TEM-1 after 1–10 mg Cl₂·L^–1 chlorination, while a UV dose of 80 mJ·cm^–2 yielded a damage ratio for bla _TEM-1 of approximately 1.2-log. Therefore, high UV doses are required for effective disruption of antibiotic resistance genes in bacteria.     

Purification and sequencing of a mate-recognition protein from the copepod<Emphasis Type="Italic"> Tigriopus japonicus</Emphasis>

Male copepods in the genus Tigriopus use surface proteins of female copepods to recognize appropriate mates. In a previous study of pre-copulatory mate-guarding in T. japonicus Mori, a monoclonal antibody was developed against one protein that contributed to the attractiveness of female T. japonicus as mate-guarding partners. In the present study, which was performed between December 1998 and November 2000, the antibody was used to purify two proteins from homogenates of pooled T. japonicus. A 70 kD protein was partially sequenced using mass spectrometry, revealing sequence similarity to α₂-macroglobulin. It was shown that the 70 kD protein was highly expressed in stage CV females, the most attractive stage for mate-guarding. A purified 36 kD protein was also shown to be a cognate of α₂-macroglobulin. Because the structure of both the 70 and 36 kD proteins suggested that protease activity and its inhibition might play a role in male mate choice, three protease inhibitors were tested in bioassays of male discrimination of conspecifics. The male's ability to discriminate among females was not completely blocked by the protease inhibitor, but their behavior was affected in more subtle ways. This suggests that the 70 kD protein, designated CSP70, is a variant of α₂-macroglobulin, but with more specific activity.     

Geographical expansion of the invader <Emphasis Type="Italic">Caprella scaura</Emphasis> (Crustacea: Amphipoda: Caprellidae) to the East Atlantic coast

Caprella scaura (Templeton in Trans Entomol Soc Lond 1:185–198, 1836) is a native species to the western Indian Ocean. It was first described from Mauritius and later reported from several regions of the world. During the last decade, the species spread out of the Adriatic Sea, and in 2005, C. scaura was reported in Gerona, on the north-eastern coast of Spain. The present study shows the recent rapid expansion of the species to the East Atlantic. During a sampling survey of harbours along the Strait of Gibraltar (Dec 2009–June 2010), we found high densities of C. scaura associated with the bryozoans Zoobotryon verticillatum Della Chiaje, 1822, in Chipiona and Bugula neritina (L.) in Cádiz, Atlantic coast of southern Spain; on the other hand, during May 2009, an important population of C. scaura was also found in the Canary Islands (Tenerife) associated with aquaculture resources. A total of 1,034 individuals of C. scaura were studied, and we confirm that the populations are effectively established, with high densities and reproducing females during the whole year. The population from Cádiz was characterised by smaller specimens but a higher number of eggs per female. These differences could be related to environmental conditions, mainly sea water temperature and to other factors such as competition with other species, or availability of food or substrate. Significant correlation was found between female size and number of eggs in the three populations, but egg size did not vary with female body length. The morphometric relation length/width of the second gnathopod was also calculated and compared between sexes: larger males presented longer gnathopods at the three sites. The most probable introduction vector of this species is shipping; in fact, the Strait of Gibraltar is characterised by an intense shipping traffic. Also, we have observed that the native Caprella equilibra Say, 1818, is being displaced by C. scaura in the harbour of Cádiz, where this species reach densities of 35,945 ind/1,000 g of B. neritina. Further experimental studies are necessary to explore the potential danger of this species, which might spread into natural habitats too.     

Intraspecific variation of sesquiterpene lactones associated to a latitudinal gradient in <Emphasis Type="Italic">Smallanthus macroscyphus</Emphasis> (Heliantheae: Asteraceae)

According to theory, variation in plant secondary metabolism against herbivores is driven by variation in biotic and abiotic conditions interacting with plants genotype to determine the expression of resistance traits. Particularly, it has been long postulated that plants growing along latitudinal gradients experience changes in biotic and abiotic interactions, specifically leading to a decrease of plant toxicity towards the poles. We tested this hypothesis using the asteraceous species Smallanthus macroscyphus. Smallanthus species are known to contain sesquiterpene lactones (STLs), bitter compounds with a broad spectrum of biological activities, including deterrence to herbivores. S. macroscyphus showed a decrease in chemical diversity of STLs when investigating populations growing from the tropical regions to less tropical ones. Populations from lower latitudes were found to be more chemically diverse with enhydrin, uvedalin and fluctuanin as main components, while populations southward were chemically fairly uniform, with polymatin A as the main and largely dominant STL. The STL chemistry of S. macroscyphus is in agreement with the hypothesis that plants of tropical forests have a greater diversity of secondary metabolites when compared to their temperate counterparts.     

Free-living marine nematodes from Biscayne Bay,Florida, I. Comesomatidae: the male of Laimella longicauda Cobb, 1920, and description of Actarjania new genus

The male of Laimella longicauda Cobb, 1920 is described as having short arcuate spicules and a gubernaculum with caudally directed apophyses. The genus Paracomesoma Schuurmans Stekhoven, 1950 is reinstated to accommodate species of Laimella sensu Chitwood, 1937. Paramesonchium n. gen. is proposed to accommodate Laimella (?) serialis Wieser, 1954. Actarjania n. gen. (type species A. splendens n. sp.) differs from Sabatieria in having irregular lateral rows of longitudinal punctations and short arcuate spicules with distinctive distal extremities.     

Seasonal changes in biomass,growth rates and production of subantarctic calanoid copepods in the Bay of Morbihan,Kerguelen Islands

The present study addresses the ecology of two dominant copepod species in the Bay of Morbihan, Kerguelen Archipelago. The biomass of the herbivore Drepanopus pectinatus (from 2 mg m^?3 in winter up to 500 mg m^?3 in summer) is tightly coupled to seasonal changes in chlorophyll a concentration in the region, whereas the biomass of the predatory euchaetiid Paraeuchaeta antarctica increases during two distinct periods over the year: 250 mg m^?3 in early summer, with the recruitment of the annual generation, and 100 mg m^?3 in autumn, with the deposition of lipids as energy reserves in C5 stages and adults. The juvenile growth rates predicted by temperature-dependent models (0.09 day^?1) closely approximate those observed in D. pectinatusin summer, but are much greater than those observed in P. antarctica (from 0.001 to 0.04 day^?1 depending on developmental stages). This difference can be explained by the reproductive strategies and trophic positions of the two species and may also result from the dependence of larval growth on energy reserves in P. antarctica. The production rates are five- and tenfold greater in juvenile stages than in adults, respectively, for D. pectinatus and P. antarctica. The secondary production by D. pectinatusis insufficient to support P. antarcticaduring winter, when the predatory species probably shifts to alternate prey. In summer the predation by P. antarctica accounts for only a minor part of the mortality estimated for D. pectinatus (from 20% to 60% depending on the examined station). At two of the three stations examined in the Bay of Morbihan, the production of P. antarctica could potentially support the dietary requirements of planktivorous seabirds in the region (~2,000 kg prey day^?1 for common diving petrels, Pelecanoides urinatrix, and ~90 kg prey day^?1 for rockhopper penguins, Eudyptes chysocome filholi).