Spatial distribution of freshwater mussels: the role of host fish and metabolic rate

SUMMARY. 1. Three species of the superfamily Unionacea live in Bavarian trout streams: the freshwater pearl mussel Margaritifera margaritifera which usually occurs alone, and the unionids Unio crassus and Anodonta piscinalis which are frequently associated.
2. We found no evidence for competition between M. margaritifera and the unionids. Two types of habitat factors were analysed: (i) the occurrence of host fish, which does not explain the distributional relationships of mussels; and (ii) hydrochemical factors indicating a lower productivity in the rivers inhabited by M. margaritifera.
3. With respect to physiological differences, M. margaritifera is separated from the other two species by low growth constants and low metabolic rates. We conclude, therefore, that the metabolic rate is an important factor governing distribution of the species: M. margaritifera is able to grow in rivers with low food availability, whereas the higher metabolic rates of U. crassus and A. piscinalis presumably necessitate a richer food supply.     

Phylogeography and conservation genetics of endangered European Margaritiferidae (Bivalvia: Unionoidea)

Margaritifera margaritifera and M. auricularia are among the most endangered freshwater mussels in the world, and the only species of the genus found in Europe. Our genetic study explores allozymic variability (27 loci) and differentiation at the mitochondrial sequence level (partial COI and 16S rRNA gene sequences). The Spanish M. auricularia population showed genetic parameters of variation that were of the same order as those of other freshwater molluscs (though at the lower end of the range), probably permitting its potential recovery. The difference between this species and M. margaritifera was clearly established (ten diagnostic allozymic loci, Nei = 0.462, and mean nucleotide divergence around 9.4%). The M. margaritifera populations analysed showed a certain degree of population genetic structure (according to allozyme data) that was not, however, related to a geographical cline. Nevertheless, two mitochondrial lineages (albeit very closely related) were identified: a northern lineage extending from Ireland to the Kola Peninsula including the western Atlantic coast, and a second cluster distributed from Ireland to the Iberian Peninsula. The phylogenetic relationships between these two species and other related taxa were established. The putative M. m. durrovensis could be considered an 'ecophenotype'. Palaeobiogeographical scenarios are presented and indicate unexpected 'recent' gene flow between M. margaritifera populations that were theoretically isolated in the early Tertiary.  © 2003 The Linnean Society of London, Biological Journal of the Linnean Society 2003, 78, 235–252.     

Freshwater Pearl mussels of the genus Margaritifera (Mollusca: Bivalvia) described as M. elongata (Lamarck, 1819) and M. borealis (Westerlund, 1871) should be classified with M. margaritifera (Linnaeus, 1758)

The shells of Pearl mussels from the basins of the Solza, Keret’, and Umba rivers flowing into the White Sea have been measured to determine the ratio of shell convexity to its maximum height. This ratio is the main character that, according to Bogatov et al. (2003), allows one to distinguish between three species of the genus Margaritifera: M. margaritifera, M. elongata, and M. borealis. It has been found that the above ratio gradually increases as the shell grows. Therefore, this character is unsuitable for species diagnosis, the more so that no hiatus in it between the three forms of pearl mussels has been revealed in any of the samples studied. On this basis, it may be concluded that Northern Europe, including Russia, is inhabited by only one species of pearl mussels, M. margaritifera.     

Translocation of wild populations: conservation implications for the genetic diversity of the black-lipped pearl oyster Pinctada margaritifera

Translocation has been widely studied as a tool for conservation management to restore or enhance degraded populations. On the contrary, few studies have been conducted on translocation for commercial purposes. In this study, we evaluate the genetic consequences of translocation of wild individuals of Pinctada margaritifera on farmed and adjacent wild populations. We tested the hypotheses that translocations would induce high genetic heterogeneity in farmed populations and this heterogeneity would then leak into the adjacent wild populations. In fact, farmed samples exhibit high levels of heterogeneity and low pairwise relatedness compared to wild populations, highlighting the pooling of genetically divergent populations into farms. We also demonstrate that this heterogeneity is transmitted to adjacent wild populations as a result of interbreeding. Adjacent wild populations tend to have higher genetic diversity values and greater pairwise relatedness coefficient with farmed populations than wild populations. Overall, pearl culture in French Polynesia promotes the mixing of unrelated individuals in farmed locations and reduces genetic divergence among geographically distant populations as well as among farmed and wild populations of a same lagoon. We also studied for the first time a farmed population originating from spat collected in a lagoon where release of hatchery-produced larvae occurred 10 years ago and we were able to identify four distinct genetic groups. These groups contribute highly to reproduction and caused considerable genetic drift in the lagoon, suggesting that hatchery-produced larvae are neither sustainable method for pearl culture nor for conserving the diversity of P. margaritifera in French Polynesia.     

Colonisation of freshwater habitats by the European eel Anguilla anguilla

1. The spatial distribution of European eels in 18 U.K. rivers was related to distance from tidal limit using a negative exponential model. This function accounted for between 19 and 90% of the variation in eel density where quantitative data was available. For semiquantitative data the negative exponential function was a significant predictor of eel densities in only six out of 10 cases, although all rivers showed a consistent decline in abundance with distance upstream from the tidal limit. 2. The spatial distribution of different age groups of European eel in River Severn showed an initial rapid dispersion into freshwater followed by a much slower dispersion rate. Movement of the population upstream by a wave‐form migration process does not occur in this system. Instead colonisation of freshwaters can be seen as a two‐phase dispersion. Phase‐1 is a rapid dispersion upstream driven by density at the point source. Phase‐2 commences once the eels become yellow eels and is equivalent to random diffusion of particles. 3. These processes have important implications for the penetration of freshwaters with reduced numbers of eel larvae arriving on the coast of Europe and North America. Eel abundance will decrease more in freshwaters in an upstream direction whilst it may remain stable or decrease to a lesser extent in estuaries. They are also able to explain the demography of eels migrating upstream over weirs and the observations of varying sex ratios within catchments. We conclude that a dispersion model dependent on age, temperature, difficulty of migration, habitat quality and density of eels should be an important part of freshwater eel management.     

Distribution of Melissococcus plutonius in Honeybee Colonies with and without Symptoms of European Foulbrood

A sensitive hemi-nested polymerase chain reaction (PCR) was used for detection of Melissococcus plutonius, the causative agent of European foulbrood (EFB). Sampling was made in Switzerland, where EFB is a widespread disease and incidences have increased in recent years. Larvae from brood samples with and without clinical signs of disease (n = 92) and honey (n = 92) from the same colonies were investigated. Individual larvae (n = 60) and pupae (n = 30) from diseased brood in single colonies were also investigated to study the distribution of the bacterium within the brood between larvae. M. plutonius was detected in larvae in all apiaries where symptoms of EFB could be seen, but not in all colonies judged as cases of EFB in the field, when healthy-looking larvae from such colonies were tested. The occurrence of the bacterium within the brood was not limited to larvae with symptoms only, but was mainly found in diseased larvae. The bacterium was also found in pupae. Healthy-looking larvae—even from heavily diseased combs—failed, in a number of cases, to amplify product in the PCR. M. plutonius could only be detected in 35% of the brood nest honey from clinically diseased colonies.     

Saline rivers provide arid landscapes with a considerable amount of biochemically valuable production of chironomid (Diptera) larvae

Saline rivers are supposed to be ‘hot spots’ of high biological productivity in arid landscapes. To test this, we quantified the production of chironomid larvae, because river production is known to be transferred to arid landscapes primarily by birds fed on these larvae. In addition, we studied the potential biochemical quality of the larvae for birds based on the essential highly unsaturated fatty acid (HUFA) contents in their biomass. We studied species composition and measured production of chironomid larvae in two saline rivers (Volgograd region, Russia). We also evaluated the fatty acid composition and contents of the dominant taxa and estimated the flux of HUFA from the studied saline rivers to land via chironomid potential emergence. Average monthly production of chironomids measured for only 1 month, August, was quite comparable to annual production in some freshwater rivers. All the dominant chironomid larvae had comparatively high essential eicosapentaenoic acid contents, especially Cricotopus salinophilus, which showed the highest value, reported for Chironomidae. The monthly flux of HUFA from the studied rivers to land due to the chironomid potential emergence was roughly comparable to the global average estimation of annual water–land HUFA export via emerging insects.     

Influence of freshwater flows on the distribution of eggs and larvae of black bream Acanthopagrus butcheri within a drought-affected estuary

This study tested the hypothesis that variable freshwater flow in the Gippsland Lakes, Australia, influences the location and extent of environmental conditions suitable for spawning and larval development of black bream, Acanthopagrus butcheri. Freshwater flow had a large influence on the salinity and level of stratification (difference between bottom and surface salinity) in the Gippsland Lakes. Freshwater flows throughout this study varied from no or low flows through to major flooding in 2007. Eggs of A. butcheri were found in similar concentrations throughout the lakes and rivers. More than 99% of larvae, however, were collected from within rivers, with very few larvae being collected from the lakes. A comparison of two spawning seasons revealed that the year with higher freshwater flows also had greater spawning activity and higher concentrations of larvae. Interestingly, there was a significant relationship between the distribution of eggs and larvae with the level of stratification. The highest concentrations of larvae occurred at sites with a difference in bottom and surface salinities of 15-20. This study demonstrates that despite A. butcheri spawning in the lakes and rivers, it is only locations with a halocline that function as larval nursery habitat.     

Effects of cryoprotectant agents and freezing protocol on motility of black-lip pearl oyster (Pinctada margaritifera L.) spermatozoa

Gamete cryopreservation techniques have been applied to several bivalve mollusc species. However, research activity in this area has primarily focused on cryopreserving gametes from edible oysters (Ostreiidae). Few studies have examined the effect of cryoprotectants and freezing protocols in the preservation of spermatozoa from cultured pearl oysters (Pteriidae). Pearl oyster producers are increasingly looking towards the development of improved family lines and, as a consequence, the ability to cryopreserve gametes would bring about significant benefits to the cultured pearl industry. In response to this need, we evaluated the effect of three cryoprotectant additives (CPA) on motility of spermatozoa from the black-lip pearl oyster, Pinctada margaritifera. These additives have previously been used to cryopreserve gametes of other bivalve species. The following CPA mixtures were evaluated: (1) 0.45M trehalose and 0, 0.64, 1.02 and 1.53 M dimethyl sulfoxide (Me(2)SO); (2) 0.2M glucose and 2M Me(2)SO and (3) 1.31 M propylene glycol (PG). The effects of four different freezing protocols on motility of P. margaritifera spermatozoa were also evaluated (slow, medium, medium-rapid and rapid cooling). This study showed that total motility was best retained when spermatozoa were cryopreserved in 0.45 M trehalose and 0, 0.64, 1.02 or 1.53 M Me(2)SO and frozen using slow to medium-rapid cooling rates (2.1-5.2 degrees Cmin(-1)). Rapid freezing through direct plunging of spermatozoa into liquid nitrogen resulted in the lowest overall retention of motility regardless of the CPA additive; however, CPA mixture also influenced retention of motility, with 0.2M glucose in 2M Me(2)SO and 1.31 M PG retaining the lowest levels of motility for the CPAs evaluated.     

Larval presence prediction through logistic regression: an early warning system against Mytilopsis leucophaeata biofouling

Mytilopsis leucophaeata is a biofouling bivalve causing major problems in the cooling water system of BASF, Antwerp NV, Belgium, a large water-using industrial facility. This study aimed to develop a statistical model to predict the response of M. leucophaeata larvae to environmental conditions in estuarine ecosystems. Multiple logistic regression, taking into account temporal autocorrelation, was applied on a large dataset allowing the prediction of the probability of occurrence of M. leucophaeata larvae at BASF NV as a response to the environmental variables. The final model made it possible to predict larval presence in the water column solely by monitoring water temperature. The results from subsampling indicated that the model was stable. The model was tested with 2005 data, demonstrating a 98% precise prediction of the occurrence of M. leucophaeata larvae in the water column, with a sensitivity of 100% and a specificity of 97%, even though autumn 2005 was exceptionally warm, which led to an extended presence of the larvae.     

Identification of calconectin, a calcium-binding protein specifically expressed by the mantle of Pinctada margaritifera

Nacre or mother-of-pearl in the shell of Pinctada margaritifera is composed of 95-99% calcium carbonate and 1-5% organic matrix. In this study, we developed an original technique to characterize the genes differentially expressed in nacre-forming cells (NFC) by combining suppression subtractive hybridization (SSH), to establish a cDNA subtractive library, with rapid amplification of cDNA ends (RACE)-PCR. Seventy-two specific cDNA sequences have been obtained so far. These include a protein containing two EF-hand Ca2+-binding domains which was completely sequenced after amplification by RACE-PCR. Its specific expression as well as the specificity of the SSH method was confirmed by semi-quantitative RT-PCR on NFC and mantle cells.     

Temperature responses in larvae of Macoma balthica from a northerly and southerly population of the European distribution range

Planktonic marine invertebrate embryos and larvae experience high mortality rates. Processes during these early vulnerable stages of development are an important determinant of the dynamics of marine invertebrate populations. In order to evaluate possible specific local adaptations of the bivalve Macoma balthica (L.), larvae from parents living in Norway (Balsfjord) and France (Gironde Estuary) were reared in the laboratory at 10, 15 and 20 °C. The rate of growth and the time it took to develop a foot were measured. Larvae grew faster and developed quicker at higher temperatures. This was true for both origins tested. Within temperature treatments, the French larvae always developed and grew fastest. Size at metamorphosis (defined as the appearance of the foot) was 250 μm (SD=12.7) in five out of the six cases; the only exception was Norwegian larvae kept at the highest temperature that metamorphosed at a smaller size (229 μm, SD=6.4). Size at metamorphosis thus appears to be largely independent of temperature. In both populations, instantaneous survival rates declined with temperature with no effect of origin. Instantaneous survival declined faster with temperature than development rates increased, resulting in lower net survival of larvae to metamorphosis at the higher temperatures. Although the French larvae had a shorter development time at the same temperature than the Norwegian larvae, the total survival of larvae from the two origins was not significantly different. The larvae of M. balthica of both populations prove to be tolerant to considerably higher rearing temperatures than they will ever experience in their natural habitat.     

Diversity of Melissococcus plutonius from honeybee larvae in Japan and experimental reproduction of European foulbrood with cultured atypical isolates

European foulbrood (EFB) is an important infectious disease of honeybee larvae, but its pathogenic mechanisms are still poorly understood. The causative agent, Melissococcus plutonius, is a fastidious organism, and microaerophilic to anaerobic conditions and the addition of potassium phosphate to culture media are required for growth. Although M. plutonius is believed to be remarkably homologous, in addition to M. plutonius isolates with typical cultural characteristics, M. plutonius-like organisms, with characteristics seemingly different from those of typical M. plutonius, have often been isolated from diseased larvae with clinical signs of EFB in Japan. Cultural and biochemical characterization of 14 M. plutonius and 19 M. plutonius-like strain/isolates revealed that, unlike typical M. plutonius strain/isolates, M. plutonius-like isolates were not fastidious, and the addition of potassium phosphate was not required for normal growth. Moreover, only M. plutonius-like isolates, but not typical M. plutonius strain/isolates, grew anaerobically on sodium phosphate-supplemented medium and aerobically on some potassium salt-supplemented media, were positive for β-glucosidase activity, hydrolyzed esculin, and produced acid from L-arabinose, D-cellobiose, and salicin. Despite the phenotypic differences, 16S rRNA gene sequence analysis and DNA-DNA hybridization demonstrated that M. plutonius-like organisms were taxonomically identical to M. plutonius. However, by pulsed-field gel electrophoresis analysis, these typical and atypical (M. plutonius-like) isolates were separately grouped into two genetically distinct clusters. Although M. plutonius is known to lose virulence quickly when cultured artificially, experimental infection of representative isolates showed that atypical M. plutonius maintained the ability to cause EFB in honeybee larvae even after cultured in vitro in laboratory media. Because the rapid decrease of virulence in cultured M. plutonius was a major impediment to elucidation of the pathogenesis of EFB, atypical M. plutonius discovered in this study will be a breakthrough in EFB research.     

Isolation and characterization of microsatellite loci in the western pearlshell mussel, Margaritifera falcata (Gould)

Ten microsatellite loci were isolated from the western pearlshell, Margaritifera falcata (Gould, 1850) and characterized in populations from Washington and Montana, USA. We also assessed eight microsatellite loci developed in M. margaritifera, two of which showed utility. Both of our test populations showed significant heterozygote deficiencies at most loci, consistent with a hermaphroditic life history. Populations differed markedly with respect to allelic richness, allele frequencies and numbers of identical multilocus genotypes. This panel of loci should prove useful in describing gene flow and genetic diversity patterns among M. falcata populations, information that should aid future conservation efforts.     

Restoration of stratum corneum with nacre lipids

To discover potential new products for the atopic dermatitis treatment, lipids extracted from nacre from the oyster Pinctada margaritifera were tested on artificially dehydrated skin explants. Expression of filaggrin and transglutaminase 1 was investigated after treatment of dehydrated skin with P. margaritifera lipid extracts according to light microscopy after labelling with specific monoclonal antibodies. The lipids were extracted from the nacre with methanol/chloroform mixture at room temperature and the extract composition was determined according to TLC and densitometry measures. Relative to the dry nacre material, a yield of extraction in lipids of 0.54% (w/w) was determined. Fatty acids, triglycerides, cholesterol and ceramides were in low abundance. Then, application of lipid formulations on skin explants previously dehydrated gave after 3 h an overexpression of filaggrin and a decrease of transglutaminase expression as shown by light microscopy. Using immunofluorescence labelling, we showed that lipids extracted from the mother of pearl of P. margaritifera induced a reconstitution of the intercellular cement of the stratum corneum. The signaling properties of the nacre lipids could be used for a development of new active product treatment against the symptoms of the dermatitis.     

Nutrient optima and tolerances of benthic invertebrates,the effects of taxonomic resolution and testing of selected metrics in lakes using an extensive European data base

A large dataset from 1,077 lakes in Finland, Ireland, Sweden and the United Kingdom was collated to analyse the relationship between nutrient status and occurrence of different taxa, as well as between total phosphorus or chlorophyll and commonly used macroinvertebrate metrics developed for river assessment. We found that most taxa were associated with mesotrophic conditions (sensu OECD). Species associated with oligotrophic status included Baetis rhodani, Gammarus lacustris and plecopteran larvae, a group commonly associated with low nutrient status also in rivers. Species tolerant of eutrophic conditions were the chironomid larvae (Chironomus plumosus and Cryptochironomus defectus); and two species of tubificids (Psammoryctides barbatus and Potamothrix hammoniensis). For a number of taxa the associations of benthic invertebrates with nutrient state reported in the literature were not supported by analysis of the REBECCA data. The analysis indicated a variable response of littoral macroinvertebrates to eutrophication pressure when using common metrics developed for macroinvertebrates in rivers. Several metrics showed significantly different responses in lakes with different alkalinity, justifying the use of alkalinity for typing water bodies. These significant responses suggest that benthic invertebrates may be a useful component for classification of ecological status in lakes. The low amount of variance explained by the regressions (<30%), however, suggests that further harmonisation of sampling methods, as well as statistically more robust assessment tools are needed to increase the comparability of datasets and to improve the precision in the dose–response relationships.     

Assessing non-target effects and host feeding of the exotic parasitoid <Emphasis Type="Italic">Apanteles taragamae</Emphasis>, a potential biological control agent of the cowpea pod borer <Emphasis Type="Italic">Maruca vitrata</Emphasis>

Apanteles taragamae Viereck is a larval parasitoid introduced in Benin for classical biological control of the cowpea pod borer Maruca vitrata Fabricius. In the laboratory, we evaluated the effects of A. taragamae on non-target herbivore species, and on another parasitoid of M. vitrata, i.e. the egg-larval parasitoid Phanerotoma leucobasis Kriechbaumer. Furthermore, we addressed the host feeding behaviour of A. taragamae. The host specificity of A. taragamae was assessed by offering six other lepidopteran species to the wasp. The competitive ability of A. taragamae was studied by providing the wasp with one- and two-days-old M. vitrata larvae that had hatched from eggs previously parasitized by P. leucobasis. Controls consisted of eggs and larvae offered only to P. leucobasis and A. taragamae, respectively. None of the other six lepidopteran species was successfully parasitized by A. taragamae. The larval parasitoid A. taragamae outcompeted the egg-larval parasitoid P. leucobasis when offered two-days-old host larvae. Competition between the two parasitoid species did not significantly affect one-day-old host larvae that were less suitable to A. taragamae. Host feeding by A. taragamae did not affect survival of one-day-old or two-days-old M. vitrata larvae. However, the percentage parasitism of two-days-old larvae was significantly reduced when exposed to female A. taragamae wasps that had been starved during 48 h. The data are discussed with regard to host specificity, host feeding patterns and to factors underlying the outcome of intrinsic competition between parasitoid species.     

Growth Biorhythms of the Freshwater Pearl Mussel Margaritifera margaritifera (Bivalvia,Margaritiferidae), Nemina River Population (Karelia)

Biology Bulletin - Individual linear growth was studied in 23 specimens of the freshwater pearl mussel Margaritifera margaritifera by measuring successive annual growth rings on the shell surface....     

Genetic analysis of threatened Australian grayling Prototroctes maraena suggests recruitment to coastal rivers from an unstructured marine larval source population

Population genetic variation of Australian grayling Prototroctes maraena was examined to determine whether the dispersal strategy of this amphidromous species favours retention of larvae and juveniles in close proximity to their natal river, or mixing of populations via marine dispersal. Variation in microsatellite and mitochondrial DNA markers was unstructured and differentiation was indistinguishable from zero across four coastal rivers spanning approximately one-quarter of the continental range of the species. This result indicates that the marine larval and juvenile phase probably facilitates extensive gene flow among coastal rivers and agrees with a previous analysis of otolith chemistry that suggested larvae probably move into the sea rather than remain in estuaries. It appears likely that the dispersal strategy of P. maraena would enable recolonization of rivers that experience localized extinction provided that connectivity between freshwater habitats and the sea is sufficient to permit migration and that enough source populations remain intact to support viability of the wider population.     

Habitat characterization of the morphologically similar mayfly larvae,Caenis and Tricorythodes (Ephemeroptera)

The larvae of Caenis (Caenidae) and Tricorythodes (Tricorythidae), once considered to be confamilial, have notable morphological and behavioural similarities. Univariate and multivariate (discriminant analysis) techniques were used to determine which environmental variables best characterized the larval habitats of Caenis and Tricorythodes at 40 sample sites on 29 rivers within the Interior Plains of Alberta, Canada. River width, depth and substrate type distinguished riverine habitats of the two genera. Larvae of Tricorythodes occurred in wide rivers of varying depths that possessed coarse substrates. Although larvae of Caenis occurred in a variety of habitats, they were found more frequently on stable substrates in narrow, deep rivers.