Indigenous microflora and opportunistic pathogens of the freshwater zebra mussel, Dreissena polymorpha

Freshwater fouling invertebrate zebra mussels (Dreissena polymorpha) harbor a diverse population of microorganisms in the Great Lakes of North America. Among the indigenous microorganisms, selective species are opportunistic pathogens to zebra mussels. Pathogenicity to zebra mussels by opportunistic bacteria isolated from the mussels was investigated in this study. Among the more than 30 bacteria isolated from temperature-stressed mussels, Aeromonas media, A. veronii, A. salmonicida subsp. salmonicida, and Shewanella putrefaciens are virulent pathogens to juvenile zebra mussels. Inoculation of a bacterial concentration of A. media, A. salmonicida subsp. salmonicida and S. putrefaciens at 10⁷ cells per zebra mussel resulted in 100% mortality within 5 days, and only 64.9% for A. veronii. In contrast, mortality was less than 12.3% following inoculation of a sterile phosphate buffer solution as a control. In addition, mortality was dependent on the size of the pathogen population used in inoculation and the incubation temperature, indicating the close relationship between the bacterial population and subsequent death. On the mussel tissue, a dense microbial population was evident from the moribund mussels viewed with Scanning Electron Microscope (SEM). Opportunistic bacteria invaded and destroyed the D. polymorpha tissue after 7 days of incubation when the bacterial inoculation was larger than 10⁵ per zebra mussel. Our results suggest that mussels are reservoirs of opportunistic pathogenic microorganisms to aquatic organisms and humans and a better understanding of the microbial ecology of the mussels will provide insights to the possible health hazards from these microorganisms.     

Bioprocessing of polluted suspended matter from the water column by the zebra mussel (Dreissena polymorpha Pallas)

Measurements of pseudofaeces production of Dreissena polymorpha were carried out with the aim of developing a biological filter at the freshwater inlet of Lake Volkerak-Zoommeer, the Netherlands. Bioprocessing of polluted suspended matter by suspended cultures of D. polymorpha occurs by filtration and sedimentation of the suspended matter as pseudofaeces. The measurements were conducted under semi-natural conditions.Pseudofaeces production was mainly determined by the dry matter content of the water; the relation is linear. Temperature was of much less importance. This agrees with earlier investigations of the filtration rate of D. polymorpha. Even at the lowest temperature measured during the experiments (6.4 °C) no large decrease in activity was observed. The relation of pseudofaeces production with shell length was sigmoid in shape, in accordance with measurements of the filtration rate.The pseudofaeces produced was slightly more polluted than suspended matter, partly due to a finer grainsize. D. polymorpha from Lake IJsselmeer exposed for 217 days at the intended location of the filter showed bioaccumulation of toxicants, especially organic pollutants and Polycyclic Aromatic Hydrocarbons (up to 10-fold accumulation). The required number of D. polymorpha in the biological filter to treat the waterflow of 14 m³ s^–1 entering Lake Volkerak-Zoommeer is 1.24 * 10⁹. The purification efficiency of the filter, the reduction of the amount of toxicants, partly depends on the binding properties of the toxicants and is highest for those strongly bound to suspended matter.     

Distribution and aspects of population dynamics of the zebra mussel,Dreissena polymorpha (Pallas, 1771), in the lake IJsselmeer area (The Netherlands)

Summary The distribution, settling and growth of the zebra mussel, Dreissena polymorpha Pallas, in Lake IJsselmeer and Lake Markermeer in The Netherlands were studied from 1980 to 1985. In these lakes D. polymorpha is the most important food source for wintering diving ducks. The study was part of an investigation into the carrying capacity of the lakes for these birds.     

Changes in the distribution and abundance of Dreissena polymorpha within lakes through time

Dreissena polymorpha population densities and biomass were followed in three Belarusian lakes with different trophic status over a 12-year period subsequent to initial colonization. In all three lakes zebra mussel population densities did not change once they reached a maximum. Application of the Ramcharan et al. [1992. Canadian Journal of Fisheries and Aquatic Sciences 49: 2611–2620] model for predicting population dynamics of zebra mussels was accurate for two of the three lakes studied. Population density appears to depend on the time since initial colonization, relative abundance of substrate available for colonization, lake morphometry and trophic type. Zebra mussel distribution within lakes was highly patchy, but the degree of dispersion decreased over time after initial colonization, which may be a result of saturation of suitable substrates by zebra mussels as populations increase and reach carrying capacity. In lakes where submerged macrophytes are the dominant substrate for zebra mussel attachment, populations may be less stable than in lakes with a variety of substrates, which will have a more balanced age distribution, and be less impacted by year to year variation in recruitment. Dreissena polymorpha usually reach maximum population density 7–12 years after initial introduction. However, the timing of initial introduction is often very difficult to determine. Both European and North American data suggest that zebra mussels reach maximum density in about 2–3 years after populations are large enough to be detected.     

Physiological energetics of the zebra musselDreissena polymorpha in lakes I. Growth and reproductive effort

Shell growth of tagged zebra mussels (Dreissena polymorpha) was monitored in lakes at 3 sites over 1 1/2 years. It varied greatly with the season and was almost absent during winter months. Shell growth was significantly correlated with seston concentration, but not with water temperature. The theoretical maximum size (L∞) of the v. Bertalanffy growth equation did not vary seasonally. Tissue weight underwent a pronounced seasonal cycle. In animals of 20 mm shell length, minimum weights recorded in September only corresponded to one quarter to one third of the maxima in late spring. Tissue weight of animals from the 3 sites was distinctly different. Carbohydrate content of the tissue stayed below 10 per cent and tended to be highest in spring. Spring maxima of lipids were very pronounced. These lipids were primarily located in the digestive gland. These data were combined with data of gonad size by Borcherding (1991). Results imply that production of reproductive tissue even continues at a retarded rate during winter months, if food conditions were favourable. With poor food conditions, no production of reproductive tissue was estimated during winter; this, however, was followed by an elevated rate of reproductive tissue production in spring. Weight specific production decreased with a weight exponent of −0.24. Shell and byssus production contributed only in the range of 10 per cent to total production. Reproductive effort showed maxima of 30 to 45%. It increased with shell length at 2 sites and decreased at the other site with the largest animals.     

The filtration rate of the zebra mussel Dreissena polymorpha used for water quality assessment in Dutch rivers

The effects of contaminated river water on the filtration rate of zebra mussels from a clean reference site were studied. After a 48-h exposure period to filtered water from the rivers Rhine, Meuse and Amstel (The Netherlands), the filtration rate was measured. It was demonstrated that water from contaminated locations inhibited the filtration rate. Inhibition was higher during low water levels in the rivers Rhine and Meuse than during high water levels, suggesting that contaminants are diluted during high water levels. It is concluded that the shortterm filtration assay with D. polymorpha can be used for assessing water quality.     

Influence of food size and food quantity on the feeding of the mussel Dreissena polymorpha

Summary In common with many other suspension feeders, the freshwater mussel Dreissena polymorpha has a maximum filtration rate at low food concentrations and a maximum ingestion rate at high food concentrations. These high rates, which reflect the potential maximum food uptake of the animal, are called the filtration capacity and the ingestion capacity respectively. The ingestion capacity was attained without forming pseudofaeces with Chlamydomonas reinhardii as food. The incipient limiting level could be calculated as the quotient of these two values. A decrease of the filtration rate at high food concentrations was correlated with changes in pumping activity, which showed more frequent interruptions, or a lower level of water transport. Dreissena can filter out particles of diameter greater than 0.7 m from the water. Retention reaches a plateau at about 5 m particle diameter. Scanning electron micrographs of the arrangement of the cilia on the gill filaments are given.     

Synchronous spawning in a recently established population of the zebra mussel,Dreissena polymorpha,in western Lake Erie,USA

Seasonal patterns of reproduction in a newly established population of Dreissena polymorpha are described for a site in the western basin of Lake Erie. Reproduction was monitored by histological examination of gonads, analysis of shell length-dry weight relationships, and following abundance of planktonic and settling larvae. Patterns of planktonic larval abundance and settling showed a distinct bimodal pattern in July and August, but conflicted with histological and length-dry weight data that showed spawning was a brief, highly synchronous event occurring in late August. Differences between histological data and abundance of larvae in the plankton can be explained as resulting from drift of larvae from disjunct populations of D. polymorpha which spawned earlier, into the study area. Veliger larvae were present in the plankton at low densities throughout the warm months before and after periods of peak abundances. The presence of these larvae can be explained by a combination of asynchronous spawning among local populations and postponed settlement by planktonic larvae.     

Size-selective predation by roach (Rutilus rutilus) on zebra mussel (Dreissena polymorpha): field stuides

Summary Studies of predation by roach (Rutilus rutilus) on zebra mussel (Dreissena polymorpha) in a large, eutrophic lake showed that there was a clearly marked size threshold ( 160 mm SL) above which roach began to feed on mussels. Roach preying on various sizes of mussels selected them in proportions different from their abundance and accessibility in the habitat. The mean size of mussels ingested by roach of 220 mm and larger, which fed predominantly on Dreissena, closely followed the pattern expected for a constant ratio of mean prey size to mean predator mouth size = 0.59. To explain the size selection we applied an optimal foraging approach, based on the ability of different-sized fish to crush (cost) mussels of different sizes, and hence crushing resistance, and energy contents (benefit). We found that fish smaller than 160 mm, which showed no inclination to eat Dreissena, would only be able to take small mussels with a very high cost/benefit ratio. The real switch to Dreissena would be expected in fish of 230–240 mm that could take most of their prey from highly profitable, numerous, and easily accessible size classes while keeping the mean prey size at the optimal level relative to mean predator mouth size.     

Physiological energetics of the zebra musselDreissena polymorpha in lakes II. Food uptake and gross growth efficiency

Food uptake ofDreissena polymorpha from two shallow water sites and one site at a 9 m depth in lakes was monitored over the course of 1 1/2 years. Feeding was characterized by the filtration capacity and ingestion capacity (i.e. the unrestricted maximum rates) at ambient temperature. The available food was estimated as seston dry weight, as seston ash free dry weight and as seston volume (sum of the volumes of the particles in the water column). The filtration capacity and ingestion capacity varied strongly during the course of the year by more than 1:10, with maximum values during late spring and early summer. The filtration capacity was correlated with water temperature, the ingestion capacity with temperature and seston concentration. However, at identical water temperatures both capacities were higher in spring than in autumn. At the 9 m site, filtration and ingestion capacity was also significantly influenced by lack of oxygen caused by stratification during summer. In the allometric equations by which both capacities (C) were related to body weightW (C=a W ^b ), an exponent b of 0.89±0.14 (mean±95% confidence interval) was calculated for the filtration capacity and 1.00±0.18 for the ingestion capacity. Both exponents did not vary significantly during the course of the year. The seasonal trend of the incipient limiting concentration (i.e. the seston concentration at which the intestine is filled to its capacity when the animal filters at maximum rate) was less pronounced. It showed a significantly positive correlation with seston concentration and a negative correlation with temperature. Except during the spring bloom, the incipient limiting concentration was in the range of the seston volume registered at the sites. Gross growth efficiencies were highly variable. An annual average between 5 and 20% was calculated. Growth efficiencies were lower at the shallow water sites with the highest seston concentrations.     

Zebra mussels (Dreissena polymorpha): a new perspective for water quality management

In the evaluation of the role of lake restoration programmesin situ measurements of the filtration rate of the freshwater musselDreissena polymorpha have been performed in Lake Wolderwijd, The Netherlands. The filtration rate mainly depends on the suspended matter content of the water, and shows an inverse exponential relationship with this factor. The filtration activity is temperature indifferent between approx. 5 and 20 °C. At low temperatures the filtration rate drops abruptly, at high temperatures gradual inhibition occurs. The filtration rate shows a sigmoidal relation with the length of the mussel. The largestD. polymorpha have a diminished filtration rate compared to animals of smaller size. This might be a degenerative feature of the oldest mussels. In Lake Wolderwijd a population density of 675 per m² is required to compensate phytoplankton growth by grazing. Manipulation of the size of the population can be executed by adding suitable substrates for the mussel.     

Genetic variability and phylogeography of the invasive zebra mussel, Dreissena polymorpha (Pallas)

The zebra mussel, Dreissena polymorpha (Pallas), a bivalve species originally native to the Black and Caspian seas, has invaded Ireland in the last decade. Five microsatellite loci were used to investigate genetic diversity and population structure in 10 populations across Europe (Ireland, UK, the Netherlands and Romania) and the Great Lakes (Lake Ontario and Lake St Clair). Levels of allelic diversity and mean expected heterozygosity were high for all populations (mean number of alleles/locus and H(E) were 10-15.2 and 0.79-0.89, respectively). High levels of polymorphism observed in Irish populations suggest that the Irish founder population(s) were large and/or several introductions took place after foundation. Significant deficits of heterozygotes were recorded for all populations, and null alleles were the most probable factor contributing to these deficits. Pairwise comparisons using Fisher exact tests and F(ST) values revealed little genetic differentiation between Irish populations. The UK sample was not significantly differentiated from the Irish samples, most probably reflecting an English origin for Irish zebra mussels. No significant differentiation was detected between the two Great Lakes populations. Our data support a northwest rather than a central or east European source for North American zebra mussels.     

Physiological energetics of the zebra musselDreissena polymorpha in lakes III. Metabolism and net growth efficiency

Oxygen consumption and ammonia excretion of the zebra musselDreissena polymorpha from 3 sites in lakes were estimated regularly over the course of 1 1/2 years at ambient temperature. They showed a pronounced annual cycle, when expressed in absolute terms (at standard shell length) and in weight specific terms (at standard tissue weight). The atomic ratio of oxygen consumed to ammonia-N released (O/N ratio) was lowest in late summer at all sites (10 to 20) and highest during winter and spring (50 to > 100). The mean body weight exponent pooled from these sites was 0.78 (95% confidence interval±0.07) for the oxygen consumption rate and 0.80 (confidence interval±0.10) for the ammonia excretion rate. Both oxygen consumption and ammonia excretion were significantly correlated with the water temperature at the 2 shallow water sites, where temperature variation was most pronounced. Correlation with seston content or gonad volume were insignificant at these sites. The quotient of filtration capacity to oxygen consumption rate was about 3 times higher at the site with the poorest food conditions compared to the other sites. Net growth efficiency was highly variable; its annual average was 35 to 40 per cent and independent of locality and animal size.     

Shoals ofDreissena polymorpha as bio-processor of seston

Mussels ofD. polymorpha were collected from mesotrophic lake Partęczyny in which it occurs in form of narrow but very dense shoal. Having transported them to the laboratory live specimens were sorted with respect to size and placed in lake water on artificial substrata made with plexi plates. During the night most of mussels reattached to substrate with byssus threads. The plates covered with mussels were transported to shallow, eutrophic lake Druzno in whichD. polymorpha has not been present in recent years. A set of plates and separate plates were exposed in lake water to which a fine fraction of sediment was added. The resulting concentration of suspension was slightly higher than those found during intensive sediment resuspension. It was observed that during first 10–30 minutes of exposure to suspension mussels kept their valves closed, but after this time they were able to continue filtration. The obtained results showed that the relatively high concentration of sediment added to lake seston did not affect clearance capacity of suspension inD. polymorpha and even enhanced it.     

Growth and reproductive cycle of the zebra mussel in the River Rhine as studied in a river bypass

In a Rhine River bypass built at a water pollution control station, the growth and reproductive cycle of the zebra mussel (Dreissena polymorpha) were studied over a period of nearly 3 years. The experimental setup offered the possibility to manipulate the temperature of the continuously passing river water and to calculate influences of temperature and food on the growth of individually marked mussels. Shell growth rates were a function of algal density (measured as chlorophyll a). This function followed saturation curves with temperature-dependent plateau levels, and saturation occurred at values above 40 μg/l chlorophyll a. The relationship between shell growth rate and temperature followed the courses of bell-shaped curves with algal-food-dependent heights. No clear correlations could be found between shell growth and other environmental factors such as dissolved organic matter, total organic content, seston, and turbidity. Two spawning periods per year characterized the reproductive cycle of the zebra mussels at the study site. The first one covered nearly 3 months (April to July) comprising several spawning events. The second shorter one occurred in August. Young mussels of the first spawning period settled at the end of May, and they were able to reproduce a few months later (end of August) when their shell length exceeded 9 mm. This first spawning period of the young-of-the-years coincided with the second spawning period of the older generations. Through the combined analysis of the results of both body weight determinations and shell growth experiments, it was possible to model mussel growth over a period of nearly 3 successive years, and to suggest the presence of an endogenous factor that might be involved in determining the start of the annual shell growth period. Received: 8 April 1997 / Accepted: 3 November 1997     

Ecotoxicity of lead to the zebra musselDreissena Polymorpha,Pallas

The effect of lead on the filtration rate of the zebra musselDreissena polymorpha was investigated, together with the accumulation of Pb in the soft tissues of the mussels. The NOEC-filtration was 116 g.l^–1 (0,56 mol.l^–1) and the EC₅₀-filtration was 370 g.l^–1 (1.79 mol.l^–1). The NOEC-accumulation was the concentration found in the control water (1.4g.l^–1). These experiments show that the EC₅₀-filtration for Pb is similar to that for Cd, higher than that for Cu and lower than that for Zn. The water quality criteria for lead allow 25 g Pb.l^–1 in surface water. This will not cause short-term effects. Long-term effects may, however, occur, since an accumulation of Pb as low as 16 g.l^–1 was recorded in this study.     

Fouling of European freshwater bivalves (Unionidae) by the invasive zebra mussel (Dreissena polymorpha)

1. The zebra mussel (Dreissena polymorpha) is well known for its invasive success and its ecological and economic impacts. Of particular concern has been the regional extinction of North American freshwater mussels (Order Unionoida) on whose exposed shells the zebra mussels settle. Surprisingly, relatively little attention has been given to the fouling of European unionoids. 2. We investigated interspecific patterns in fouling at six United Kingdom localities between 1998 and 2008. To quantify the effect on two pan‐European unionoids (Anodonta anatina and Unio pictorum), we used two measures of physiological status: tissue mass : shell mass and tissue glycogen content. 3. The proportion of fouled mussels increased between 1998 and 2008, reflecting the recent, rapid increase in zebra mussels in the U.K. Anodonta anatina was consistently more heavily fouled than U. pictorum and had a greater surface area of shell exposed in the water column. 4. Fouled mussels had a lower physiological condition than unfouled mussels. Unlike tissue mass : shell mass ratio, tissue glycogen content was independent of mussel size, making it a particularly useful measure of condition. Unio pictorum showed a stronger decline in glycogen with increasing zebra mussel load, but had a broadly higher condition than A. anatina at the time of study (July). 5. Given the high conservation status and important ecological roles of unionoids, the increased spatial distribution and fouling rates by D. polymorpha in Europe should receive more attention.     

Expression pattern of vasa in gonads of sea cucumber Apostichopus japonicus during gametogenesis and reproductive cycle

The vasa gene is a reliable germline marker to study the origin and development of germ cells and gonads, although the gene product (mRNA or protein) varies between different species. However, there has been little study on vasa genes in holothuroids to date. Here we determined the expression characteristics of the Apostichopus japonicus vasa gene (Aj-vasa) during gametogenesis in the ovary and testis using in situ hybridization and immunohistochemistry. During oogenesis, the expression pattern of Aj-vasa coincided at the mRNA and protein levels. Intensive signals in oogonia decreased gradually with the development of oocytes. Interestingly, the pattern was different during spermatogenesis. The Aj-vasa mRNA level was the highest in spermatogonia, reduced in spermatocytes, low in spermatids and absent in spermatozoa, but the Aj-VASA protein was restricted to spermatogonia and early spermatocytes. These expression characteristics of Aj-vasa persisted in both male and female gonads throughout the reproductive cycle. Our findings show that Aj-vasa mRNA is a good marker for studying the origin and migration of germline cells; moreover, Aj-VASA is a useful tool to identify spermatogonia in A. japonicus. Our findings indicate that Aj-vasa is vital in the development and differentiation of germ cells.     

The zebra mussel (Dreissena polymorpha), a new pest in North America: reproductive mechanisms as possible targets of control strategies

Summary

The zebra mussel (Dreissena polymorpha) has spread rapidly in temperate fresh waters of North America since its introduction into the Great Lakes in 1985 or 1986. It attaches to hard substrates, forming layers, occluding water intakes, encrusting and killing native mussels, filtering algae in competition with other planktivores, and possibly interfering with fish spawning. It reproduces prolifically, suggesting that an approach to its control may be by controlling its reproduction. Previous literature suggests that spawning in bivalves is regulated by both environmental and internal chemical cues. A suggested sequence is that phytoplankton chemicals initially trigger spawning; chemicals associated with gametes provide a species-specific pheromonal positive feedback for spawning; and the response to environmental chemicals is mediated internally by serotonin (5-HT). The role of 5-HT in zebra mussels is under investigation. Both males and females can be induced to spawn by either injection or external application of 5-HT. The response can also be activated by 8-hydroxy-2-(di-n-propylamino)-tetralin, an agonist at 5-HT_1A receptors. HPLC analysis has detected 5-HT as the major biogenic amine in both male and female gonads. 5-HT immunocytochemistry demonstrates nerves containing serotonergic fibers innervating gonads of both males and females, with prominent varicosities surrounding the follicles in both sexes. A role of 5-HT in mediating spawning responses in zebra mussels is thus strongly supported. These studies have shown that reproductive behavior of zebra mussels can be modified by outside chemicals, a property that may be exploited for purposes of control.     

Detection of DNA damage in haemocytes of zebra mussel using comet assay

The aim of the study was to use the comet assay on haemocytes of freshwater mussel, Dreissena polymorpha Pallas, for detection of possible DNA damage after exposure to pentachlorophenol (PCP) and to evaluate the potential application of the comet assay on mussel haemocytes for genotoxicity monitoring of freshwater environment. Zebra mussels were exposed for seven days to different concentrations (10, 80, 100, 150 μg/l) of PCP and in the river Sava downstream from Zagreb municipal wastewater outlet. Significant increase in DNA damage was observed after exposure to PCP at doses of 80 μg/l and higher and after in situ exposure in the river Sava as well. This study confirmed that the comet assay applied on zebra mussel haemocytes may be a useful tool in determining the potential genotoxicity of water pollutants.