Molecular characterization and evolution of haemocyanin from the two freshwater shrimps Caridina multidentata (Stimpson, 1860) and Atyopsis moluccensis (De Haan, 1849)

Haemocyanin (Hc) is a copper-containing respiratory protein, floating freely dissolved in the hemolymph of many arthropod species. A typical haemocyanin is a hexamer or oligohexamer of six identical or similar subunits, with a molecular mass around 75 kDa each. In the crustaceans, the haemocyanins appear to be restricted to the remipedes and the malacostracans. We have investigated the haemocyanins of two freshwater shrimps, the Amano shrimp Caridina multidentata and the bamboo shrimp Atyopsis moluccensis. We obtained three full-length and one partial cDNA sequences of haemocyanin subunits from the Amano shrimp, which were assigned to the α- and γ-types of decapod haemocyanin subunits. Three complete and two partial haemocyanin cDNA sequences were obtained from the bamboo shrimp, which represent subunit types α, β and γ. This is the first time that sequences of all three subunit types of the decapod haemocyanins were obtained from a single species. However, mass spectrometry analyses identified only α- and γ-type subunits, suggesting that a β-subunit is not a major component of the native haemocyanin of the bamboo shrimp. Phylogenetic and molecular clock analyses showed that malacostracan haemocyanins commenced to diversify into distinct subunit types already ~515 million years ago. β-subunits diverged first, followed by α- and γ-type subunits ~396 million years ago. The haemocyanins of phyllocarids and peracarids form distinct clades within the α/γ-cluster. Within the Caridea, an early divergence of distinct α-type subunits occurred ~200 MYA. The tree of the γ-subunits suggests a common clade of the Caridea (shrimps) and Penaeidae (prawns).     

A molecular phylogenetic study of the freshwater talitrid amphipod Orchestia cavimana (Crustacea).

In this paper we performed a molecular phylogenetic study of Orchestia cavimana, the sole talitrid amphipod inhabiting beaches of European freshwater lakes and rivers. For that purpose, we have PCR amplified and sequenced regions of the mitochondrial cytochrome oxidase subunit I (COI) gene, basing our analysis on both nucleotide and amino acid sequences and considering also structural classes of the COI enzyme. Phylogenetic analyses were conducted by neighbour-joining (NJ) and maximum-parsimony (MP) methods comparing homologous sequences of talitrids and other Crustacea. In both NJ and MP trees, O. cavimana shows a basal placement with respect to other talitrid amphipods.     

Haemocyanin oxygen binding and the physiological ecology of a range of talitroidean amphipods (Crustacea) II. Effect of freezing,inorganic ions,and urate on O2 binding in vitro

Summary The effect of variations in [K], [Ca], [Mg], [NaCl], and [urate] on the in vitro O₂ binding properties of haemocyanin (Hc) from three talitroidean species, viz. the aquatic Apohyale pugettensis, the semi-terrestrial Megalorchestia californiana, and the semi-/euterrestrial Traskorchestia traskiana were studied. Freezing altered the cooperativity of Hc from A. pugettensis and M. californiana but not T. traskiana. Variations in [NaCl], [K], and [Mg] had no effect on cither O₂ affinity or cooperativity of the Hc except for A. pugettensis Hc where an increase in [Mg] resulted in an increase in both O₂ affinity and cooperativity. Increasing [Ca] or [urate] increased O₂ affinity of both A. pugettensis and M. californiana but not T. traskiana Hc. These effects were most marked in A. pugettensis. The results suggest a negative correlation between sensitivity to Hc effectors and the degree of terrestrial adaptation of a particular amphipod species.Abbreviations Hc haemocyanin - STR Stock traskorchestia ringer     

Studies on habitat selection by amphipods using artificial substrates within an estuarine environment

Field studies on habitat selection and community development of four gammarid amphipod species were conducted in a brackish lagoon at Nanakita River Estuary, Japan. Development of amphipod community was studied by focussing mainly on provision of shelter for these invertebrates. Seven artificial substrates (including wire net cage, circular concrete block, dried oyster shell, brush, sponge, blue and green tapes) were used in the field as substitutes of natural substrates (macroalgae). This paper demonstrates that amphipods will mostly select and inhabit substrates with available space that provide them with adequate protection. Brush substrate was ranked the best among the selected substrates, because its filamentous nature like natural substrates (macroalgae) provided suitable habitat for both tube-dwelling and free-living amphipods and accommodated maximum densities of amphipods in all seasons at all stations. Tube-dwelling amphipods (domicolous) constructed their tubes in and on algal substrates and their densities tended to be correlated with algal biomass, whilst free-living species (nestlers) prowled on surfaces, clang to filamentous structures and hid themselves in crevices, holes and boulders. Brush substrates at different stations suggested the importance of site difference.     

On the potential of fatty acids as trophic markers in Arctic grazers: feeding experiments with sea urchins and amphipods fed nine diets of macroalgae

To elucidate dietary preferences of benthic grazers at the Arctic Kongsfjorden by means of fatty acid trophic markers, ten different parallel treatments (starvation, 3 species of red macroalgae, 4 species of brown macroalgae and 2 species of green macroalgae) were offered as mono-algal diet to specimens of the dominant sea urchin Strongylocentrotus droebachiensis (Echinodermata, Echinoidea) and the gammarid amphipod Gammarellus homari (Crustacea, Amphipoda). At the end of the 3-week feeding experiments, amphipods and sea urchins (soft tissue) were deep-frozen and analysed for total lipid contents as well as fatty acid (FA) compositions. In addition, FA profiles of the algal species were determined and screened for specific FA patterns or single FAs qualifying as potential trophic markers in the grazers. Despite their diets of nine algal species with different FA compositions, FA patterns of the sea urchins and amphipods revealed a pronounced similarity between treatments. This strong similarity was also observed in the faecal pellets of the sea urchins. Hence, deviating FA compositions of the macroalgae were neither reflected in the FA patterns of the grazers’ tissue nor in their faecal pellets. Suitable algal FA trophic markers could thus not be identified in the two grazers from Kongsfjorden. The rather low lipid levels, especially in the amphipods, as well as a pronounced degradation and modification of FAs may explain that the FA trophic marker approach did not provide evidence of dietary preferences. Future experiments may obtain a higher resolution of potential FA trophic markers by analysing separate lipid classes or single tissues of lipid-poor grazers. Alternatively, different methods are needed to reveal high-resolution trophic relationships between macroalgae and herbivores in Kongsfjorden.     

Host manipulation of a freshwater crustacean (Gammarus roeseli) by an acanthocephalan parasite (Polymorphus minutus) in a biological invasion context

Several gammarid species serve as intermediate hosts for the acanthocephalan parasite Polymorphus minutus. This parasite influences gammarid behaviour in order to favour transmission to its ultimate host, generally a bird. We investigated this host manipulation in Gammarus roeseli, a gammarid species introduced in France 150 years ago which now coexists with several exotic species from different origins. In the field, vertical distribution of G. roeseli revealed a higher proportion of infected individuals close to the water's surface and the size distribution of infected gammarids revealed predation pressure on infected individuals. However, under laboratory conditions both infected and non-infected individuals remained benthic. The addition of a second gammarid, Dikerogammarus villosus, to the experimental device involved a vertical displacement of infected G. roeseli towards the water's surface. Dikerogammarus villosus, originating from the Ponto-Caspian basin, can be considered as an aggressive predator. The substitution of D. villosus with Atyaephyra desmarestii, a planktivore decapod, did not alter the gammarids' distribution, with both infected and uninfected G. roeseli staying benthic. Thus, biotic interactions between D. villosus and G. roeseli represent selective pressure encouraging the expression of manipulated behaviour in infected amphipods. Through manipulation, P. minutus was found to increase the survival of infected G. roeseli when faced with non-host predators and to make it more vulnerable to predation by the parasite's definitive host.     

STUDIES OF MESOHERBIVORY IN AQUARIA AND IN AN UNBARRICADED MARICULTURE FARM ON THE CHINESE COAST1

The effects upon macrophytic and epiphytic algae of caprellid amphipods, including Caprella irregularis Mayer, and of gammarid amphipods, including Ampithoe lacertosa Bate, Jassa falcata (Montagu) and Pontogeneia rostrata Gurjanova, were studied in aquaria and in a coastal mariculture farm near Qingdao, People's Republic of China. The macrophytic species studied included Enteromorpha linza (L.) J. Agardh, Gracilaria asiatica Zhang & Xia, Gracilaria lemaneiformis (Bory) Weber-van Bosse and Scytosiphon lomentaria (Lyngbye) C. Agardh. These algae and amphipods were selected for study because of their local abundances. Aquarium experiments demonstrated that C. irregularis, J. falcata and P. rostrata feed on ephemeral/epiphytic algae. A. lacertosa (adults and juveniles) ate Gracilaria but preferred E. linza. The different species of amphipods had substantially different patterns of daily activity. Caprellids prevented epiphytic overgrowth of G. asiatica when added to plants in an experimental, openocean, mariculture farm, despite the presence of numerous amphipod predators (e.g. the fishes Hexagrammos otakii Jordan & Starks and Sebastes thompsoni Jordan & Hubbs). Periodic removals of all (naturally colonizing) amphipods from another group of plants in the farm resulted in heavy epiphytic overgrowth. Since these results were obtained in an unbarricaded mariculture farm, we conclude that mesoherbivores may have large effects upon algal community structure in temperate intertidal and subtidal habitats.     

Structural basis for salinity-induced alteration in oxygen binding by haemocyanin from the estuarine amphipod Chaetogammarus marinus (L.)

In the estuarine amphipod Chaetogammarus marinus, differences in O(2) binding by haemocyanins (Hc) could be related to natural and salinity-related quantitative variation in just one polypeptide subunit (band 2) and not to variations in any of the other seven bands present. Band 2 was always present, irrespective of salinity treatment, and naturally makes up 6%-36% of the total Hc. However, low salinity exposure (S=4 per thousand for 48-49 h, T=15 degrees C) was accompanied by an increase in the prevalence of greater concentrations of band 2 (i.e., range, 20%-36% of total Hc present) and a concomitant increase in Hc-O(2) affinity (half saturation [P(50)] decreased from 1.38 to 1.12 kPa at pH=7.81, T=15 degrees C). A similar salinity-related mechanism (that is, one band altering) has been shown previously for the blue crab Callinectes sapidus, although the functional consequences were different. In contrast with C. marinus, an increase in the proportion of one polypeptide subunit in C. sapidus resulted in a decrease in Hc-O(2) affinity. This study has confirmed that between-individual variation (quantitative rather than qualitative) in just one Hc subunit may have functional consequences, although the significance of such variation is difficult to interpret.     

BRIEF COMMUNICATIONS Persistence times of four amphipod species in the stomachs of brown trout

Four amphipod species fed to brown trout Salmo trutta persisted in an easily identifiable form in trout stomachs for similar lengths of time. The data also allowed estimation of the maximum length of time ingested amphipods could persist in an identifiable form in brown trout stomachs at different temperatures. The data are thus useful in assessment of fish feeding patterns on amphipod assemblages in the laboratory and field.     

Accumulation and exchange of parasites during adaptive radiation in an ancient lake

In the ancient Lake Baikal, Russia, amphipod crustaceans have undergone a spectacular adaptive radiation, resulting in a diverse community of species. A survey of microsporidian parasites inhabiting endemic and non-endemic amphipod host species at the margins of Lake Baikal indicates that the endemic amphipods harbour many microsporidian parasite groups associated with amphipods elsewhere in Eurasia. While these parasites may have undergone a degree of adaptive radiation within the lake, there is little evidence of host specificity. Furthermore, a lack of reciprocal monophyly indicates that exchanges of microsporidia between Baikalian and non-Baikalian hosts have occurred frequently in the past and may be ongoing. Conversely, limitations to parasite exchange between Baikalian and non-Baikalian host populations at the margins of the lake are implied by differences in parasite prevalence and lack of shared microsporidian haplotypes between the two host communities. While amphipod hosts have speciated sympatrically within Lake Baikal, the parasites appear instead to have accumulated, moving into the lake from external amphipod populations on multiple occasions to exploit the large and diverse community of endemic amphipods in Lake Baikal.     

中国淡水钩虾一新种(甲壳纲:端足目:钩虾科)

记述了采自四川九寨沟自然保护区的淡水钩虾1新种,四川钩虾Gammarus sichuanensisi sp.nov.新种的形态特征和与近似种的区别在文中均有详细的论述,同时附有详细的特征图,模式标本保存于中国科学院动物研究所。     

Patterns of gammaridean amphipod abundance and species composition associated with dominant subtidal macroalgae from the western Antarctic Peninsula

The communities of gammaridean amphipods associated with eight dominant macroalgal species were examined near Palmer Station, Western Antarctic Peninsula. A total of 78,415 individuals belonging to 32 amphipod taxa were identified with mean densities ranging up to 20 individuals/g algal wet wt. The most abundant amphipod taxon, Metaleptamphopus pectinatus, was found to associate predominately with the brown alga Desmarestia menziesii, while the second most common taxon, Jassa spp. occurred primarily on the red alga Gigartina skottsbergii. Non-metric multidimensional scaling analysis demonstrated that the population densities of each amphipod species and amphipod species composition were similar on the same algal species but dissimilar on different species of algae. Comparisons of amphipod communities associated with a given algal species but from different sampling sites indicated that although the structure of species-specific macroalgal-associated amphipod communities can vary across spatial scales of 3 km, 50% of the macroalgal species examined showed no significant inter-site differences in associated amphipod community structure. Spearman rank correlation analyses showed that higher abundances of amphipods occurred on the macroalgae with the highest number of branches. As many Antarctic amphipods are known consumers of macroalgae, their remarkable abundances are likely to play a significant role in mediating energy and nutrient transfer in nearshore Antarctic Peninsular macroalgal communities.     

Patterns of population genetic variation in sympatric chiltoniid amphipods within a calcrete aquifer reveal a dynamic subterranean environment

Calcrete aquifers from the Yilgarn region of arid central Western Australia contain an assemblage of obligate groundwater invertebrate species that are each endemic to single aquifers. Fine-scale phylogeographic and population genetic analyses of three sympatric and independently derived species of amphipod (Chiltoniidae) were carried out to determine whether there were common patterns of population genetic structure or evidence for past geographic isolation of populations within a single calcrete aquifer. Genetic diversity in amphipod mitochondrial DNA (cytochrome c oxidase subunit I gene) and allozymes were examined across a 3.5 km² region of the Sturt Meadows calcrete, which contains a grid of 115 bore holes (=wells). Stygobiont amphipods were found to have high levels of mitochondrial haplotype diversity coupled with low nucleotide diversity. Mitochondrial phylogeographic structuring was found between haplogroups for one of the chiltoniid species, which also showed population structuring for nuclear markers. Signatures of population expansion in two of the three species, match previous findings for diving beetles at the same site, indicating that the system is dynamic. We propose isolation of populations in refugia within the calcrete, followed by expansion events, as the most likely source of intraspecific genetic diversity, due to changes in water level influencing gene flow across the calcrete.     

Gill Na(+), K(+)-ATPase in a series of hyper-regulating gammarid amphipods: enzyme characterisation and the effects of salinity acclimation

The enzyme Na(+), K(+)-ATPase was investigated in the gills of selected hyper-regulating gammarid amphipods. Gill Na(+), K(+)-ATPase was characterised with respect to the main cation and co-factor concentrations for the freshwater amphipod Gammarus pulex. The optimum cation and co-factor concentrations for maximal gill Na(+), K(+)-ATPase activity in G. pulex were 100mM Na(+), 15mM K(+), 15mM Mg(2+) and 5mM ATP, at pH 7.2. The effects of salinity acclimation on gill Na(+), K(+)-ATPase activity and haemolymph sodium concentrations was investigated in selected gammarid amphipods from different salinity environments. Maximal enzyme activity occurred in all gammarids when acclimated to the most dilute media. This maximal activity coincided with the largest sodium gradient between the haemolymph and the external media. As the haemolymph/medium sodium gradient decreased, a concomitant reduction in gill Na(+), K(+)-ATPase activity occurred. This implicates the involvement of gill Na(+), K(+)-ATPase in the active uptake of sodium from dilute media in hyper-regulating gammarids.     

Effects of Acid Mine Drainage on the Estuarine Food Web, Britannia Beach, Howe Sound, British Columbia, Canada

We investigated the effects of acid mine drainage (AMD) from Britannia Creek, BC, Canada, a stream contaminated by an abandoned copper mine, on the estuarine food web. Near the mouth of Britannia Creek, sediments and the water contained high concentrations of dissolved copper and the waters were highly acidic. Relative to a reference location, rockweed (Fucus gardneriSilva) cover was reduced, phytoplankton biomass was lower, chironomid larvae were less abundant, and fewer gammarid amphipods colonized basket traps. Laboratory toxicity tests confirmed that sediments and water from the lower reaches of Britannia Creek and its estuary were highly toxic to midge larvae (Chironomus tentansFabricius), the marine amphipod (Eohaustorius washingtonianusThorsteinson) and also impaired fertilization of sand dollars (Dendraster excentricusEschscholtz). Analyses of stomach contents of chum salmon fry (Oncorhynchus ketaWalbaum) indicated that they preferred to feed on chironomid larvae and gammarid amphipods at Britannia Creek estuary but these two taxa were less abundant at Britannia Creek estuary relative to the reference location. AMD from Britannia Mine disrupted the structure and function of the intertidal ecosystem resulting in loss of food production for fish including chum salmon fry and other species.     

Island endemism, morphological stasis, and possible cryptic speciation in two coral reef, commensal Leucothoid amphipod species throughout Florida and the Caribbean

Coral reefs are believed to be one of the most diverse ecosystems, but the true magnitude of their biodiversity and patterns of endemism is uncertain. This uncertainty stems partly from the relative paucity of investigations on the small, difficult to collect taxa (cryptofauna) that may make up the majority of reef biodiversity and require specialized expertise for morphological identification. To assess the extent of diversity in some of the reef micro-cryptofauna, we analyzed 414 bp of the mitochondrial cytochrome oxidase subunit I gene from 556 individuals representing two brooding amphipod species (Leucothoe ashleyae and Leucothoe kensleyi). These amphipods are commensal inside the branching vase sponge Callyspongia vaginalis, and were sampled throughout Florida and the Caribbean. Phylogenetic analyses revealed 11 deeply divergent, strongly supported lineages (seven L. ashleyae and four L. kensleyi) each with very narrow geographic ranges. The level of intraspecific lineage divergence for both morphospecies was among the highest reported for any marine crustacean (12.4–26.0% uncorrected), and exceeded that of congeners from nine diverse amphipod families, as well as the patristic genetic distance suggested as a threshold for crustacean species delineation. These findings suggest a history of cryptic speciation within each morphospecies, concomitant with a pronounced period of morphological stasis involving each of the morphotypes. The observation of multiple, highly divergent, evolutionary significant units, each endemic to Florida and Caribbean island locations, supports the emerging view that coral reef biodiversity, especially in the cryptofaunal component, is likely vastly underestimated.     

Effects of height on the shore and complexity of habitat on abundances of amphipods on rocky shores in New South Wales, Australia

Patterns of abundance of epifaunal crustaceans were compared between two common brown algae in intertidal rock pools on a shore near Sydney, New South Wales, Australia. Amphipods were significantly more abundant on Sargassum globulariaefolium (which was most common low on the shore) than on Hormosira banksii (more common at mid-shore levels). Experiments tested the hypotheses that height on the shore and structural complexity of the algae were the main factors influencing patterns of abundance of amphipods. Reciprocal transplants using defaunated plants and similar experiments using artificial plants allowed simultaneous tests of these hypotheses. The abundance of a common intertidal amphipod, Sunampithoe graxon was significantly smaller on Sargassum transferred from low to mid-tidal level of the shore than on any other treatments, indicating that height on the shore affects numbers of this species. There were no Sunampithoe on Hormosira even when transplanted to low shore areas, indicating that other factors, apart from the height on the shore, are also important for this amphipod. Another common amphipod, Hyale maroubrae was generally found in greatest abundances on Hormosira transplanted from mid- to low shore and on Sargassum low on the shore, again indicating the importance of height on the shore. There was no difference in abundance of amphipods on ‘complex’ and ‘simple’ artificial plants indicating that structural complexity (as defined in this study) did not influence these amphipods.     

Feeding rates of common Antarctic gammarid amphipods on ecologically important sympatric macroalgae

Single species feeding trials employing both fresh algal tissues and alginate food pellets containing dried finely ground algal tissues were conducted to examine the relative palatability of sympatric Antarctic macroalgae (three brown and five red macroalgal species) to three common herbivorous gammarid amphipods (Prostebbingia gracilis Chevreux, Gondogeneia antarctica (Chevreux) Thurston, and Metaleptamphopus pectinatus Chevreux). In fresh algal tissue bioassays, both the amphipods P. gracilis and G. antarctica consumed significantly greater amounts of the red alga Palmaria decipiens over all other seven species of macroalgae. The amphipod M. pectinatus failed to consume measurable quantities of fresh thalli of any macroalgae and therefore is likely to feed on other resources. In food pellet bioassays, the consumption rates of amphipods fed with eight different species of macroalgae were compared with consumption rates on a highly palatable control green alga. Alginate pellets containing finely ground tissues of P. decipiens were consistently the most palatable of any of the macroalgae to P. gracilis and G. antarctica, while pellets containing the brown algae Desmarestia menziesii, D. anceps and the red alga Plocamium cartilagineum were not consumed by any of the three amphipod species. Regression analysis indicated that feeding rates of the amphipods P. gracilis and G. antarctica on alginate food pellets were not significantly correlated with known species-specific parameters of macroalgal nutritional quality (%N, %C, C:N ratio, soluble protein, soluble carbohydrate, and lipid). Therefore, differences in amphipod macroalgal palatability are most likely related to other factors including physical and/or chemical deterrents.     

Seasonal variation in trace metal concentrations in three talitrid amphipods from the Gulf of Gdansk, Poland

Talitrid amphipod crustaceans are increasingly being used as biomonitors of trace metal bioavailabilities in coastal waters, and it is important to understand how other factors, in addition to metal pollution, might affect their accumulated body concentrations. Seasonal variation (April-October 2000) in body concentrations of copper, zinc, cadmium, iron, manganese, lead and nickel has been investigated in three species, Orchestia cavimana, Talitrus saltator and Talorchestia deshayesii, from the region of the Gulf of Gdansk (including the Vistula Lagoon), Poland, an area with considerable temperature variation between summer and winter. Seasonal variation in the body metal concentrations of talitrid amphipod crustaceans was confirmed in each population, but only copper showed a consistent pattern. Copper concentrations fell from early to late summer, perhaps either as a result of the replacement of the older overwintering generation by the younger spring one, or as a result of the loss of haemocyanin in autumn amphipods entering a period of low metabolic activity. Metal concentration data for T. saltator from Gdynia-Osada Rybacka compared with equivalent data for 1996 and 1998 show a significant drop in metals associated with the influx of the Vistula river water, which occurred in 1998, following a major flood event in 1997.