Influence of cortisol on the attachment and metamorphosis of larval Utterbackia imbecillis on bluegill sunfish (Lepomis macrochirus)

The larvae of unionid freshwater mussels (i.e., glochidia) undergo a parasitic stage requiring their attachment to the external epithelia of fish hosts, where they metamorphose into free-living juveniles. We describe the physiological effects in bluegill sunfish (Lepomis macrochirus) of infection with glochidia from the paper pondshell (Utterbackia imbecillis). Glochidia accumulation on bluegill increased dramatically at concentrations of 2000 glochidia liter(-1) and above, reaching a maximum attachment density of about 30 glochidia g(-1) fish at 4000 glochidia liter(-1). Plasma cortisol was the most sensitive indicator of biological effect to glochidial exposure, increasing significantly in hosts exposed to 2000 glochidia liter(-1) or greater. Glochidia were 31% more likely to undergo successful juvenile metamorphosis when attached to bluegill with elevated plasma cortisol, largely due to the enhanced survivorship of these larvae during the first 48 h after infection. We tested the hypothesis that glochidial attachment and juvenile metamorphosis were stimulated directly by plasma cortisol in fish hosts. Bluegill were given an intraperitoneal injection of cortisol, then infected with 1000 glochidia liter(-1) at 48 h after hormone supplementation. Cortisol-injected fish had a 42% increase in the number of attached glochidia g(-1) fish and a 28% increase in larval metamorphosis compared to sham-injected and control fish. We provide evidence that cortisol enhances glochidial metamorphosis on hosts by improving the retention of attached glochidia. This study gives insights into the influence of host physiology on glochidial attachment and juvenile mussel transformation.     

Physiological and ecological hosts of Popenaias popeii (Bivalvia: Unionidae): laboratory studies identify more hosts than field studies

1. Freshwater mussels are critically endangered in North America, making it important to understand their environmental requirements at all life stages. As glochidia (larvae), they attach to fish hosts where they undergo substantial mortality, making this transition important in their life cycle. Larval host fish requirements have typically been described using data from laboratory infestations to determine suitable hosts. 2. Laboratory infestations circumvent many natural barriers that prevent infestation of physiologically compatible fishes by mussel larvae. While such methods are invaluable for identifying ‘physiological hosts,’ they cannot fully describe realised ‘ecological hosts’ in the field. 3. We studied Popenaias popeii in the Black River in New Mexico, because it is of conservation concern and it is the only mussel species present, facilitating identification of glochidial infestation. To explore the difference between physiological hosts and ecological hosts, we conducted a 3‐year field study of fishes infested by P. popeii glochidia. 4. Substantially fewer fish species were infested by P. popeii in the wild (10 of 20 observed) than had been identified as physiological hosts in laboratory trials (24 of 31). We combined data on fish abundance, proportion of fish hosts infested (prevalence) and the number of glochidia per fish (intensity) and identified three fish species that probably contributed substantially more to mussel recruitment by carrying more glochidia than other host species. 5. Similarities in behaviour among these fishes allowed us to hypothesise routes of infestation, such as benthos‐feeding by catostomids, that allow glochidia to infest these hosts at higher rates than other suitable hosts. Overall, this approach provides a method of quantifying the relative importance of different species of host fish in the mussel lifecycle.     

Site selection and attachment duration of Anodonta woodiana (Bivalvia: Unionacea) glochidia on fish hosts

The relationship between parasitic glochidia of Anodonia woodiana (Unionidae: Anodonti-nae) and potential fish hosts was investigated in the laboratory. Intensity of parasitism was highest on the exotic fish Gambusia affinis , lower on the native species Puntius semijascio-latus and Metzia takakii , and least on Rhodeus sinensis . Glochidia generally attached at fin margins, particularly the pectorals and caudal. In Gambusia affinis , the incidence of glochidia on the pectorals was higher than would be expected on the basis of ratios between fin margin lengths. Apparently, the role of the pectoral fins in locomotion makes them more liable to glochidial contact, thereby increasing their susceptibility to attachment.
The duration of glochidial attachment was shortened as temperatures increased. Mean values ranged from 14-4 days at 15°C to 6 days at 27°C. At 33°C glochidia rapidly detached and metamorphosis was unsuccessful. Significantly, water temperatures in A. woodiana habitats in Hong Kong rarely exceed 30°C.     

Evolution of bilaterally asymmetrical larvae in freshwater mussels (Bivalvia: Unionoida: Unionidae)

Bilaterally asymmetrical glochidia (i.e. bivalved parasitic larvae bearing a large marginal appendage on a single valve) have been reported from five Asian freshwater mussel genera belonging to two separate subfamilies, the Gonideinae (i.e. Pseudodon, Solenaia, and Physunio) and Rectidentinae (i.e. Contradens and Trapezoideus). This classification requires that the bilaterally asymmetrical glochidium‐bearing mussels are not monophyletic, and suggests that this atypical larval morphology evolved twice in the same geographic region. Although homoplastic glochidium characters are known (e.g. marginal appendages and size), we hypothesized that bilaterally asymmetrical glochidia represent a novel morphological synapomorphy. We tested the monophyly of the mussels bearing bilaterally asymmetrical glochidia using a molecular matrix consisting of representatives from all six freshwater mussel families and three molecular markers (28S, 16S, and COI). Bayesian inference, maximum likelihood, and ancestral state reconstruction were employed to estimate the phylogeny and larval trait transformations. The reconstructed phylogeny rejects the monophyly of the asymmetrical glochidium‐bearing mussels and resolves two putative origins of asymmetrical glochidia; however, ancestral state reconstruction supports asymmetrical glochidia as a synapomorphy of only one supraspecific taxon of the Rectidentinae. In the Gonideinae, asymmetrical glochidia were autapomorphic of Pseudodon cambodjensis (Petit, 1865). That is, no other taxa resolved among the Gonideinae had bilaterally asymmetrical glochidia, including other Pseudodon species. We describe how the alleged intrageneric glochidial variation in Pseudodon, and in the other genera of the Gonideinae reported to have asymmetrical glochidia (i.e. Solenaia and Physunio), challenge the resolved convergence of asymmetrical glochidia. Our results are discussed in the context of freshwater mussel larval evolution, patterns in life‐history traits, and the classification of freshwater mussels generally. © 2015 The Linnean Society of London     

Acquired resistance of bluegill sunfish Lepomis macrochirus to glochidia larvae of the freshwater mussel Utterbackia imbecillis (Bivalvia: Unionidae) after multiple infections

The effects of multiple infections on the host-parasite relationship between bluegill sunfish (Lepomis macrochirus) and parasitic glochidial larvae of the freshwater mussel Utterbackia imbecillis were examined. Na?ve, young-of-the-year bluegills were infected with glochidia and placed in individual observation chambers. Each day, water was drained from each chamber and the numbers of dead glochidia, live glochidia, partially metamorphosed glochidia, and fully metamorphosed juvenile mussels were counted. The same fishes were infected a total of 4 times. After 2 infections, the fish began to exhibit evidence of acquired resistance to glochidia. During the third and fourth infections, this resistance was clearly evidenced by the marked increase in the percentage of dead and live glochidia shed during the first 5 days of the infection and by the significant decrease in the success of metamorphosis. The total number of glochidia that successfully attached to the fish decreased significantly during the fourth infection relative to the first. The number of larvae attached to the host fish was positively correlated with the size of the fish during the first infection but was negatively correlated during all subsequent infections. Variance to mean ratios indicated that larvae were aggregated among host fishes during the infections. This study has important implications in propagation and conservation efforts of this endangered group of organisms.     

Effects of increasing temperatures on population dynamics of the zebra mussel <Emphasis Type="Italic">Dreissena polymorpha</Emphasis>: implications from an individual-based model

Zebra mussels (Dreissena polymorpha, Pallas, 1771) have had unprecedented success in colonizing European and North American waters under strongly differing temperature regimes. Thus, the mussel is an excellent model of a species which is able to cope with increasing water temperatures expected under global change. We study three principle scenarios for successful survival of the mussel under rising temperatures: (1) no adaptation to future thermal conditions is needed, existing performance is great enough; (2) a shift (adaptation) towards higher temperatures is required; or (3) a broadening of the range of tolerated temperatures (adaptation) is needed. We developed a stochastic individual-based model which describes the demographic growth of D. polymorpha to determine which of the alternative scenarios might enable future survival. It is a day-degree model which is determined by ambient water temperature. Daily temperatures are generated based on long-term data of the River Rhine. Predictions under temperature conditions as recently observed for this river that are made for the phenology of reproduction, the age distribution and the shell length distribution conform with field observations. Our simulations show that temporal patterns in the life cycle of the mussel will be altered under rising temperatures. In all scenarios spawning started earlier in the year and the total reproductive output of a population was dominated by the events later in the spawning period. For maximum temperatures between 20 and 26°C no thermal adaptation of the mussel is required. No extinctions and stable age distributions over generations were observed in scenario 2 for all maximum temperatures studied. In contrast, no population with a fixed range of tolerated temperatures survived in scenario 3 with high maximum temperatures (28, 30, 32°C). Age distributions showed an excess of 0+ individuals which resulted in an extinction of the population for several thermal ranges investigated. Priority programme of the German Research Foundation—contribution 14.     

珠蚌科六种蚌的钩介幼虫形态比较研究

本文观察了背角无齿蚌、沼纹冠蚌、剑状矛蚌、圆顶珠蚌、刻裂丽蚌和洞穴丽蚌等６种蚌的育儿囊类型;用光镜和扫描电镜观察和研究了幼虫的形态特征。结果表明,前４种蚌的育儿囊属外鳃类的同生型,后２种的育儿囊属外鳃类的四生型;６种幼虫均为钩介幼虫,其中前４种为有钩型,后２种为无钩型;６种幼虫的大小、形态有别,并证明种间幼虫的壳钧和壳饰的亚显微形态显然有异。本文还就６种蚌在Ｓｉｍｐｓｏｎ分类系统中的地位与按它们的育儿囊和钩介幼虫类型得到的归类进行比较。结果表明,Ｓｉｍｐｓｏｎ对６种蚌中３种的分类与归类不妥。本文为研究我国淡水贝类的分类提供一条新的途径,并为建立珠蚌类的自然分类系统积累资料。     

Factors influencing the upper temperature tolerances of three mussel species in a brackish water canal: size, season and laboratory protocols

Mussels are the most problematic organisms encountered in the water intake systems of electrical power plants. Various fouling control measures are adopted, among which heat treatment is considered the relatively more attractive from economic and ecological points of view. Thermal tolerance experiments were carried out to determine the effects of mussel size (2-20 mm shell length), season (breeding vs non-breeding), nutritional status (fed vs non-fed), acclimation temperature (5-25 degrees C) and acclimation salinity (1-35%o) on the mortality pattern of three important mussel species, viz. a freshwater mussel Dreissena polymorpha, a brackish water mussel Mytilopsis leucophaeata and a marine mussel Mytilus edulis under different temperatures (36-41 degrees C). The mussels in the 10 mm size group exposed to 36 degrees C showed 100% mortality after 38 min (D. polymorpha), 84 min (M. edulis) and 213 min (M. leucophaeata). The effect of mussel size on M. edulis and M. leucophaeata mortality at different temperatures was significant, with the largest size group of mussels showing greater resistance, while no significant size-dependence was observed in the case of D. polymorpha. All the three mussel species collected during the non-breeding season (June-October). Nutritional status had no significant influence on the thermal tolerance of the three mussels; fed and non-fed mussels showed 100% mortality at comparable rates. Acclimation temperature had a significant effect on the mortality of all three species. Survival time at any given target temperature increased with increasing acclimation temperature. The acclimation salinity showed no significant effect on the thermal tolerance of the three mussel species. In comparison, M. leucophaeata was more tolerant to high temperature stress than the other two species. The present studies clearly show that various factors can influence the mortality of D. polymorpha, M. edulis and M. leucophaeata to elevated temperatures. The results, therefore, suggest that if heat treatment were to be used as a control measure for these mussels, it has to be employed judiciously, depending on the mussel species, mussel size, breeding season, water temperature and salinity.     

Habitat traits, population structure and host specificity of the freshwater pearl mussel Margaritifera margaritifera in the Waldaist River (Upper Austria)

In European streams and rivers, the freshwater pearl mussel (Margaritifera margaritifera L., 1758) faces extinction. This is also true for the Waldaist River, with 20,000 specimens recorded in the early 1990s then Austria’s most important pearl mussel river. Nowadays, there is only a single 320 m stretch with noteworthy mussel densities. During an in-depth survey of this river stretch in 2010, we detected a total of 2,774 specimens. Mussel microhabitats were confined to patches of sand and fine gravel (psammal and akal) at run sections of the river, stabilized by large boulders. Pearl mussels avoided large accumulations of fine sediments. Typically situated at undercut slopes, preferred microhabitats were 0.25–0.50 m deep at baseflow with current velocities (at 40% depth) of 0.2–0.6 m s^?1. A comparison of the present stock with data from 1997 revealed a rapid decline in mussel density down to 27%. We also noticed strongly reduced growth and a high mortality of medium age classes. Juvenile mussels were completely lacking. With respect to host specificity in terms of glochidia survival, the brook char Salvelinus fontinalis (Mitchill, 1814), a suitable host in North America, shed glochidia within eight days. In the brown trout Salmo trutta L., 1758, two strains were investigated. Glochidia survival, growth and prevalence were significantly higher in the Danish than in the Austrian hatchery strain.     

A delayed effect of the aquatic parasite <Emphasis Type="Italic">Margaritifera laevis</Emphasis> on the growth of the salmonid host fish <Emphasis Type="Italic">Oncorhynchus masou masou</Emphasis>

Parasitic species often have detrimental effects on host growth and survival. The larvae of the genus Margaritifera (Bivalvia), called glochidia, are specialist parasites of salmonid fishes. Previous studies have reported negligible influences of the parasite on their salmonid hosts at natural infection levels. However, those studies focused mainly on their instantaneous effects (i.e., during the parasitic period). Given the time lag between physiological and somatic responses to pathogen infections, the effect of glochidial infection may become clearer during the post-parasitic period. Here, we examined whether the effect of glochidial infections of Margaritifera laevis on its salmonid host Oncorhynchus masou masou would emerge during the post-parasitic period. We performed a controlled aquarium experiment and monitored fish growth at two time intervals (i.e., parasitic and post-parasitic periods) to test this hypothesis. Consistent with previous observations, the effects of glochidial infection were unclear in the middle of the experiment (day 50; parasitic period). However, even with a natural glochidial load (48 glochidia per fish), we found a significant reduction in growth rates of infected fish in the extended period of the experiment (day 70; post-parasitic period). Our results suggest that examining only instantaneous effects may provide misleading conclusions about mussel–host relationships.     

Timing of spawning and glochidial release in Scottish freshwater pearl mussel (Margaritifera margaritifera) populations

1. The timing of reproduction was investigated in six Scottish freshwater pearl mussel populations from 1993 to 2002. Gravid females were examined and the release of mussel larvae (glochidia) was monitored. 2. Annual spawning (oviposition) and spat (glochidial release) events occurred during June to July and June to September, respectively. 3. Between‐river differences in timing seem to be related to water temperature. Mussels in the warmest rivers tend to spawn and spat first, and vice‐versa. 4. Thermal variations also seem to influence the timing of reproduction within rivers, which can be delayed by several weeks during cold years. At least 3000°‐days occur between annual episodes of glochidial release. 5. The timing of spawning is determined gradually, probably by a thermal summation effect. 6. The release stage occurs as a sudden, synchronised event, with most of the glochidia spat over 1–2 days, indicating that it is triggered by an environmental cue. Sudden changes in water temperature and/or river level often result in spats, and the underlying mechanism may be respiratory.     

Practical experience in the rearing of freshwater pearl mussels (Margaritifera margaritifera): advantages of a work-saving infection approach,survival, and growth of early life stages

The critically endangered freshwater pearl mussel (Margaritifera margaritifera Linnaeus 1758) is the target species of an Austrian conservation project that involves captive breeding. In order to optimize the operational procedure, controls were conducted at several decisive stages, including infection of host fish (for which a time- and work-saving enclosure approach was tested), larval growth during the parasitic stage, growth of juvenile mussels in climate chambers at different temperatures, and growth and survival of re-introduced juveniles in field cages. High infection rates could be attained under near natural conditions. Distinctive patterns in the way the gill arches of the host fish were infected could be detected. Encysted glochidia showed significantly different successive growth stages, related to water temperature. In all, five distinctive growth stages could be detected in the course of the first 562 days of observation. The stages are described and the respective daily increments given. Very high survival rates were achieved during hibernation in the field as well as at the laboratory. The study suggests a way for saving time in the infection procedure that can more effectively be invested in an intensive maintenance of juveniles at the laboratory and during hibernation in the field.     

The pathology and seawater performance of farmed Atlantic salmon infected with glochidia of Margaritifera margaritifera

The pathology of glochidial infection of the freshwater mussel Margaritifera margaritifera was examined in farmed Atlantic salmon Salmo salar in fresh water and for 150 days after transfer of salmon to sea water. Prevalence of infection in fresh water was 95%, mean abundance 134 per fish and mean infection intensity 140. Prevalence in sea water was 80–94% in the first 7 weeks after transfer but glochidia were absent, apart from remains, after 50 days in sea water. Glochidia on salmon in fresh water were associated with localized hyperplasia and fusion of secondary gill lamellae. Focally extensive epithelial hyperplasia and fusion of secondary lamellae were present 4–10 days after transfer to sea water. Twenty-three days after transfer, small nodules with a more discrete appearance were present suggesting partial resolution of tissue response; hyperplastic responses associated with glochidia were much reduced after 50 days. Plasma chloride in infected fish 10 days after transfer was 153 mmol. 1⁻¹, significantly higher than fish without infection, suggesting poorer adaptation to sea water. No mortalities due to glochidial infection in sea water were recorded and there was no significant difference in mean weight between infected and control fish.     

Persistence of host response against glochidia larvae in Micropterus salmoides

Host fish acquire resistance to the parasitic larvae (glochidia) of freshwater mussels (Unionidae). Glochidia metamorphose into juvenile mussels while encysted on host fish. We investigated the duration of acquired resistance of largemouth bass, Micropterus salmoides (Lacepède, 1802) to glochidia of the broken rays mussel, Lampsilis reeveiana (Call, 1887). Fish received three successive priming infections with glochidia to induce an immune response. Primed fish were held at 22-23 degrees C and were challenged (re-infected) at intervals after priming. Metamorphosis success was quantified as the percent of attached glochidia that metamorphosed to the juvenile stage and were recovered alive. Metamorphosis success at 3, 7, and 12 months after priming was significantly lower on primed fish (26%, 40%, and 68% respectively) than on control fish (85%, 93%, and 92% respectively). A second group of largemouth bass was similarly primed and blood was extracted. Immunoblotting was used to detect host serum antibodies to L. reeveiana glochidia proteins. Serum antibodies were evident in primed fish, but not in naive control fish. Acquired resistance of host fish potentially affects natural reproduction and artificial propagation of unionids, many of which are of conservation concern.     

The role of fecundity and reproductive effort in defining life‐history strategies of North American freshwater mussels

Selection is expected to optimize reproductive investment resulting in characteristic trade‐offs among traits such as brood size, offspring size, somatic maintenance, and lifespan; relative patterns of energy allocation to these functions are important in defining life‐history strategies. Freshwater mussels are a diverse and imperiled component of aquatic ecosystems, but little is known about their life‐history strategies, particularly patterns of fecundity and reproductive effort. Because mussels have an unusual life cycle in which larvae (glochidia) are obligate parasites on fishes, differences in host relationships are expected to influence patterns of reproductive output among species. I investigated fecundity and reproductive effort (RE) and their relationships to other life‐history traits for a taxonomically broad cross section of North American mussel diversity. Annual fecundity of North American mussel species spans nearly four orders of magnitude, ranging from < 2000 to 10 million, but most species have considerably lower fecundity than previous generalizations, which portrayed the group as having uniformly high fecundity (e.g. > 200000). Estimates of RE also were highly variable, ranging among species from 0.06 to 25.4%. Median fecundity and RE differed among phylogenetic groups, but patterns for these two traits differed in several ways. For example, the tribe Anodontini had relatively low median fecundity but had the highest RE of any group. Within and among species, body size was a strong predictor of fecundity and explained a high percentage of variation in fecundity among species. Fecundity showed little relationship to other life‐history traits including glochidial size, lifespan, brooding strategies, or host strategies. The only apparent trade‐off evident among these traits was the extraordinarily high fecundity of Leptodea, Margaritifera, and Truncilla, which may come at a cost of greatly reduced glochidial size; there was no relationship between fecundity and glochidial size for the remaining 61 species in the dataset. In contrast to fecundity, RE showed evidence of a strong trade‐off with lifespan, which was negatively related to RE. The raw number of glochidia produced may be determined primarily by physical and energetic constraints rather than selection for optimal output based on differences in host strategies or other traits. By integrating traits such as body size, glochidial size, and fecundity, RE appears more useful in defining mussel life‐history strategies. Combined with trade‐offs between other traits such as growth, lifespan, and age at maturity, differences in RE among species depict a broad continuum of divergent strategies ranging from strongly r‐selected species (e.g. tribe Anodontini and some Lampsilini) to K‐selected species (e.g. tribes Pleurobemini and Quadrulini; family Margaritiferidae). Future studies of reproductive effort in an environmental and life‐history context will be useful for understanding the explosive radiation of this group of animals in North America and will aid in the development of effective conservation strategies.     

Guild structure in water mites (Unionicola spp.) inhabiting freshwater mussels: choice,competitive exclusion and sex

Summary Unionicolid water mites inhabit freshwater unionid mussels during the nymphal and adult stages of their life-cycle. Regular sampling of mussels from two sites in St. Mark's River, Fl. established that each of four species of water mite (Unionicola abnormipes, U. fossulata, U. serrata and U. formosa) occurred mainly in one or two of the mussel species available at each site.The role of preference for particular mussel species during host location was assessed for the first three mite species by choice experiments, in which mites were offered different mussel species simultaneously. In five out of six experiments, mites entered normally unused mussels as often as they did normally used ones. Additionally, a sexual difference in choice was found for U. fossulata, with males preferring one mussel species and females showing no preference. One mussel species, (Anodonta imbecilis), normally unused but chosen by mite species during the lab. experiments, is inhabited exclusively by the fourth mite species, U. formosa, in the field. An experiment showed that U. formosa excludes other mite species aggressively from Anodonta imbecilis.The results illustrate the sometimes misleading nature of simple sampling data as an indication of host specificity or host preference in parasites. They suggest also that the population dynamics of some parasites might be more fruitfully compared to unrelated, free-living species than to other parasites.     

Experimental measurements of zebra mussel (Dreissena polymorpha) impacts on phytoplankton community composition

1 . To investigate direct effects of zebra mussel ( Dreissena polymorpha ) feeding activities on phytoplankton community composition, short‐term microcosm experiments were performed in natural water with complex phytoplankton communities. Both gross effects (without resuspension of mussel excretions) and net effects (with resuspension) were studied.
2. Gross clearance rates were not selective; essentially all taxa were removed at similar rates ranging from 24 to 63 mL mussel^–1 h^–1. Net clearance rates were highly selective; different plankton taxa were removed at very different rates, ranging from 12 to 83% of the gross rates, leading to consistent changes in the phytoplankton community composition. Thus, although zebra mussels can cause most phytoplankton to decline, there is considerable variation among taxa in either pre‐digestive selection or post‐digestive survival.
3. The direct, short‐term effects of zebra mussels on phytoplankton community composition are consistent with some of the major changes observed in the Hudson River since establishment of zebra mussels.
4. We show, with simple calculations, how zebra mussel filtration rate, its selective efficiency on various taxa, and phytoplankton growth rates interact to produce changes in the phytoplankton composition.     

Mussel responses to flood pulse frequency: the importance of local habitat

1. Understanding mechanisms behind the distribution of organisms along a gradient of hydrological connectivity is crucial for sustainable management of river–floodplain systems. We tested the hypothesis that frequency of flood pulses exerts a direct influence on the distribution of freshwater mussels (Unionoida) by creating a local environment that limits their fitness. 2. Multiscale habitat analyses combined with transplant‐rearing experiments were carried out with a focus on abundance, presence/absence, survival rates and growth rates of mussels. Sixty‐nine floodplain waterbodies (FWBs) were surveyed within a 15‐km lowland segment of the Kiso River in Japan. 3. The abundance of mussels significantly increased with increased frequency of inundation associated with flood pulses at the among‐FWB scale, while the probability of occurrence of mussels was negatively predicted by the amount of benthic organic matter at the within‐FWB scale. 4. Field‐rearing experiments showed that survival rates were low and growth rates nearly zero in infrequently inundated FWBs (these FWBs had no naturally occurring resident mussels). In such FWBs, hypoxia (DO < 2 mg L^?1) was frequently observed near the bottom when temperature was optimal for mussel growth (>15 °C). 5. These findings demonstrated that flood pulse frequency was the most important factor in determining mussel distribution in FWBs because it directly limits mussels’ fitness by mediating local environmental factors, possibly dissolved oxygen (DO) levels. Successful restoration efforts for mussel habitat conservation should focus on processes that lead to improved local conditions.     

Effects of temperature on baseline and genotoxicant-induced DNA damage in haemocytes of Dreissena polymorpha

The potential application of the Comet assay for monitoring genotoxicity in the freshwater mussel Dreissena polymorpha was explored and a preliminary investigation was undertaken of the baseline levels of DNA damage in mussel haemocytes of animals kept at different temperatures. In addition, in vitro cell sensitivity against genotoxicants was assessed in relation to increasing temperatures. The mussels were kept at four different constant temperatures (4, 18, 28 and 37 degrees C) for 15 h. The haemocytes withdrawn were treated in vitro with melphalan, as a model genotoxic compound, or sodium hypochlorite, a common water disinfectant capable of producing mutagenic/carcinogenic by-products, at the established temperatures for 1h. The data obtained in vivo, in cells directly withdrawn from the mussels showed a significant (P<0.001, Student's t test) inter-individual variability, probably due to genetic and epigenetic factors and an increasing amount of DNA damage at increasing temperature. Mussel haemocytes showed a clear dose-response effect after in vitro melphalan treatment. Hypochlorite treatment also significantly increased DNA migration: the damage was temperature dependent, with a similar increase at 4 and 28 degrees C and a minimum level at 18 degrees C. This study demonstrates the potential application of the Comet assay to haemocytes of D. polymorpha. However, these findings suggest that temperature could alter both DNA damage baseline levels in untreated animals and cell sensitivity towards environmental pollutants in in vitro conditions. Therefore, more information is needed about seasonal variations and the natural background levels of DNA damage in mussels living in the wild, before they are used for the monitoring of genotoxic effects in aquatic environments.     

The reproductive biology of the depressed river mussel, Pseudanodonta complanata (Bivalvia: Unionidae), with implications for its conservation

Pseudanodonta complanata is listed as ‘Near Threatened’on the IUCN Red List (IUCN, 2006) and is a species of conservationpriority on the UK Biodiversity Action Plan. The UK is hostto some of the largest populations of this species, but littleis known about their reproductive biology. Two populations werestudied in the Great Ouse catchment and the Waveney and Yarecatchment, East Anglia, UK. Both populations are reproductivelyactive, producing viable glochidia. Small mussels (<30 mmlength) in both catchments indicate that recruitment is occurring.A short non-gravid period in May is followed by three monthsof glochidial formation in June, July and August; by Septembermussels contain mature glochidia ready for release the followingApril. Females brood between 5,000 and 50,000 glochidia, andthis scales with mussel length L (mm) as: n_glochL^2.1. The sexratio is skewed towards females (2.5 females:1 male), and malesare larger than females. No hermaphrodites were found duringthe histological examination of the gonads of 24 mussels. Thein vivo examination of demibranchs is shown to be an effectivenonsacrificial means of determining sex and gravidity. Conservationrecommendations include: minimizing management operations inriver stretches containing large populations; avoiding formsof management which preferentially remove large mussels (i.e.males and the most fecund females); performing management duringthe non-gravid period to avoid causing the premature releaseof glochidia; leaving sufficient time between management operationsfor populations to recover; and temporarily translocating musselsto refugia during management operations. (Received 9 February 2005; accepted 2 May 2007)