Impact of anthropogenic stressors on the population genetics of the common bioindicator Dreissena polymorpha (Pallas, 1771)

Toxicant inputs from agriculture, industry and human settlements have been shown to severely affect freshwater ecosystems. Pollution can lead to changes in population genetic patterns through various genetic and stochastic processes. In my thesis, I investigated the impact of anthropogenic stressors on the population genetics of the zebra mussel Dreissena polymorpha. In order to analyze the genetics of zebra mussel populations, I isolated five new highly polymorphic microsatellite loci. Out of those and other already existing microsatellite markers for this species, I established a robust marker set of six microsatellite loci for D. polymorpha. rnMonitoring the biogeographical background is an important requirement when integrating population genetic measures into ecotoxicological studies. I analyzed the biogeographical background of eleven populations in a section of the River Danube (in Hungary and Croatia) and some of its tributaries, and another population in the River Rhine as genetic outgroup. Moreover, I measured abiotic water parameters at the sampling sites and analyzed if they were correlated with the genetic parameters of the populations. The genetic differentiation was basically consistent with the overall biogeographical history of the populations in the study region. However, the genetic diversity of the populations was not influenced by the geographical distance between the populations, but by the environmental factors oxygen and temperature and also by other unidentified factors. I found strong evidence that genetic adaptation of zebra mussel populations to local habitat conditions had influenced the genetic constitution of the populations. Moreover, by establishing the biogeographical baseline of molecular variance in the study area, I laid the foundation for interpreting population genetic results in ecotoxicological experiments in this region.rnIn a cooperation project with the Department of Zoology of the University of Zagreb, I elaborated an integrated approach in biomonitoring with D. polymorpha by combining the analysis techniques of microsatellite analysis, Comet assay and micronucleus test (MNT). This approach was applied in a case study on freshwater contamination by an effluent of a wastewater treatment plant (WWTP) in the River Drava (Croatia) and a complementary laboratory experiment. I assessed and compared the genetic status of two zebra mussel populations from a contaminated and a reference site. Microsatellite analysis suggested that the contaminated population had undergone a genetic bottleneck, caused by random genetic drift and selection, whereas a bottleneck was not detected in the reference population. The Comet assay did not indicate any difference in DNA damage between the two populations, but MNT revealed that the contaminated population had an increased percentage of micronuclei in hemocytes in comparison to the reference population. The laboratory experiment with mussels exposed to municipal wastewater revealed that mussels from the contaminated site had a lower percentage of tail DNA and a higher percentage of micronuclei than the reference population. These differences between populations were probably caused by an overall decreased fitness of mussels from the contaminated site due to genetic drift and by an enhanced DNA repair mechanism due to adaptation to pollution in the source habitat. Overall, the combination of the three biomarkers provided sufficient information on the impact of both treated and non-treated municipal wastewater on the genetics of zebra mussels at different levels of biological organization.rnIn my thesis, I could show that the newly established marker set of six microsatellite loci provided reliable and informative data for population genetic analyses of D. polymorpha. The adaptation of the analyzed zebra mussel populations to the local conditions of their habitat had a strong influence on their genetic constitution. We found evidence that the different genetic constitutions of two populations had influenced the outcome of our ecotoxicological experiment. Overall, the integrated approach in biomonitoring gave comprehensive information about the impact of both treated and non-treated municipal wastewater on the genetics of zebra mussels at different levels of biological organization and was well practicable in a first case study.

Application of a computational model for complex fluvial ecosystems: The population dynamics of zebra mussel Dreissena polymorpha as a case study

The potential and adaptive flexibility of population dynamic P-systems (PDP) to study population dynamics suggests that they may be suitable for modelling complex fluvial ecosystems, characterized by a composition of dynamic habitats with many variables that interact simultaneously. Using as a model a reservoir occupied by the zebra mussel Dreissena polymorpha, we designed a computational model based on P systems to study the population dynamics of larvae, in order to evaluate management actions to control or eradicate this invasive species. The population dynamics of this species was simulated under different scenarios ranging from the absence of water flow change to a weekly variation with different flow rates, to the actual hydrodynamic situation of an intermediate flow rate. Our results show that PDP models can be very useful tools to model complex, partially desynchronized, processes that work in parallel. This allows the study of complex hydroecological processes such as the one presented, where reproductive cycles, temperature and water dynamics are involved in the desynchronization of the population dynamics both, within areas and among them. The results obtained may be useful in the management of other reservoirs with similar hydrodynamic situations in which the presence of this invasive species has been documented.

Assessment of Ametryn Contamination in River Water, River Sediment, and Mollusk Bivalves in Sao Paulo State, Brazil

Sao Paulo state, Brazil, is one of the main areas of sugar cane agriculture in the world. Herbicides, in particular, ametryn, are extensively used in this extensive area, which implies that this herbicide is present in the environment and can contaminate the surface water by running off. Thereby, residues of ametryn were analyzed in samples of river water an river sediment and in freshwater bivalves obtained from the rivers Sapucai, Pardo and Mogi-Guacu in Sao Paulo State, Brazil. Samples were taken in the winter of 2003 and 2004 in two locations in each river. The specimens of freshwater bivalves collected and analyzed were Corbicula fluminea, an exotic species, and Diplodon fontaineanus, a native species. Additionally, the evaluation of the ability of bioconcentration and depuration of ametryn by the freshwater bivalve Corbicula fluminea was also performed. Ametryn concentrations in the samples were measured by liquid chromatography coupled to mass spectrometry. Residues of ametryn in water (50 ng/L) and in freshwater bivalves (2-7 ng/g) were found in the Mogi-Guacu River in 2004, and residues in river sediments were found in all rivers in 2003 and 2004 (0.5-2 ng/g). The observation of the aquatic environment through the analysis of these matrixes, water, sediment, and bivalves, revealed the importance of the river sediment in the accumulation of the herbicide ametryn, which can contaminate the biota.

Trends in fish prey capture by common terns following zebra mussel invasion : do terns track fish species abundance? /

The Common Tern (Sterna hirundo) is a ground nesting colonial seabird. Terns rely primarily on small prey fishes which they obtain through plunge diving for their survival as well as the survival of their offspring during the breeding season. The zebra mussel (Dreissena polymorpha) is a small bivalve mollusk that invaded North American waters in the late 1980's. Through its suspension feeding, the zebra mussel has the ability to alter the entire aquatic ecosystem, ultimately leading to a reduction in pelagic organisms including small prey fish. The objective of the study was to determine what (if any) indirect effects the invasion of the zebra mussel has had on fish prey captured by terns. The study took place in two separate two-year periods, 1990-91 and 1995-96 on a concrete breakwall off the north shore of Lake Erie near Port Colborne, Ontario. Daily nest checks revealed clutch initiation dates, egg-laying chronology, hatching success and morphological egg characteristics (length and breadth). Behavioural observations included time each sex spent in attendance with its brood, the frequency of feeding chicks and the prey species composition and size fed to chicks as well as to females (courtship feeding). Egg sizes did not differ between study periods, nor did feeding rates to chicks, suggesting that food was not a limiting resource. Terns spent less time with their broods (more time foraging) in the 1995-96 period. However, they also had significantly larger broods and fledged more offspring. The time of each individual foraging trip decreased, suggesting that fish were easier to obtain in 1995 and 1996. Lastly, kleptoparasitism rates decreased, suggesting that the costs of foraging (time, energy) actually decreased as fewer birds adopted this strategy to compensate for what I assumed to be a lack of available food (fish). The only significant difference between the periods of 1990, 1991 and 1995, 1996 was a change in diet. Terns delivered significantly fewer rainbow smelt and more emerald shiner in 1995 and 1996. However, the average size of fish delivered did not change. Thus, there was little impact on prey captured by Common Terns in Lake Erie since the invasion of the zebra mussel.

Environmental assessment and biomonitoring of the Twelve Mile Creek watershed, Niagara Peninsula, Ontario

In light of the heavy reliance of the people of the Niagara Peninsula on the T\\'elve Mile Creek (TMC) watershed for recreational activities and for municipal and industrial uses ( e.g., drinking water, shipping and discharge of effluents), it was deemed prudent to assess the envirol1tnental health of the system by analysing the sediments total and exchangeable metal, and TPH contents. The MOEE has set guidelines with limits for the protection and management of aquatic sediments, and the sediments from the headwaters of the TMC have total metal and TPH (subset of O&G) contents well below the lower provincial limits. Areas of environmental concern where total metal contents in sediments, either individually or collectively, exceed the guideline, are the south side of Lake Gibson, the Old WeIland Canal, a segment of TMC just south of the QEW and Martindale Pond. The total metal content of sediments does not in all instances identify areas of biological concern. Instead, it has been found that the exchangeable metal fraction of sediments is a better indicator of metal availability and thus potential accumulation in organisms. In some instances, the exchangeable metal fraction agrees with the total metal fraction defining areas of environmental concern, but it does vary from site to site reflecting the natural variability of the ambient environment. Overall, the exchangeable metal fraction of sediments appears to be a better indicator of anthropogenic pollution and ecosystem impact. A histochemical study of Anodon.ta sp., Elliptio sp. and zebra mussels (Dreissena polyn'101pha) was done in conjunction with passive biomonitoring of zebra and quagga mussels (Dreissena bugensis) from the Twelve Mile Creek watershed and Lake 51. Clair (Jeanette's Creek, Chatham, Ontario). The highest concentrations of divalent metals such as Cu, Ni, Cd, and Zn, and trivalent Al appear to accumulate in gill and kidney tissues. Metal contents of organ tissues in Anodonta sp. vary with size class. Organ metal content varies among size classes, thus requiring consideration of size in biomonitoring studies. Shucked zebra and quagga mussel tissues, exhibited similar size class to Al content trends. In addition they reflected the Al content trends of top (approximately 10 cm) most sediments in the Twelve Mile Creek watershed. Quagga mussels appear to have higher Al concentrations than zebra mussels, thus suggesting that quagga mussels may be better passive biomonitors of AI. Cd content in zebra mussel tissues, seemed to increase with size class trends. This was not demonstrated in the quagga mussel tissues. This suggests that Cd may be regulated by quagga mussels and not by zebra mussels, and that zebra mussels may be better passivebiomonitors of Cd than are quagga mussels. Zebra mussel, quagga mussel, Anodonta sp., and Elliptio sp. were used in a two part, active (translocated) biomonitoring study of the Twelve Mile Creek watershed. There was no statistical difference in death rates between zebra and quagga mussels after 65 days of biomonitoring. However there does appear to be a difference of death rates between sites. Unfortunately the data base did not permit us to differentiate between sites. Relative to Port Colborne Harbour (Port Colborne, Ontario), the Twelve Mile Creek watershed appears to be elevated in bioavailable AI. An area near the terminus of the Twelve Mile Creek appears to be an area of environmental concern since mussels seemed to have accumulated relatively large concentrations of Cd, Zn, and Pb. In addition to possible metal loading from a nearby outfalls, or possible upstream outfalls, road salt runoff from storm sewers may have contributed to metal accumulation through cation exchanges processes. Similar trends in cumulative quagga mussel metal concentrations during the two time periods (65 and 159 days), suggest that quagga mussels may reach equilibrium within 65 days of translocation. Differences in bioaccumulated metal concentrations of the two dreissenid species demonstrate that active biomonitoring studies must use a variety of organisms to adequately assess the environmental situation of specific waterways and/or bodies.

Auswirkungen des Klimawandels auf die klein- und großräumige genetische Populationsstruktur von Makrozoobenthos in Rhein, Main und Mosel

In den letzten Jahrzehnten wurde eine deutliche, anhaltende Veränderung des globalen Klimas beobachtet, die in Zukunft zu einer Erhöhung der durchschnittlichen Oberflächentemperatur, erhöhten Niederschlagsmengen und anderen gravierenden Umweltveränderungen führen wird (IPCC 2001). Der Klimawandel wird in Flüssen sowohl mehr Extremereignisse verursachen als auch das Abflussregime bisher schmelzwasserdominierter Flüsse zu grundwassergespeisten hin ändern; dies gilt insbesondere für den Rhein (MIDDELKOOP et al. 2001). Um die möglichen Auswirkungen dieser Veränderungen auf die genetische Populationsstruktur von Makrozoobenthosorganismen vorhersagen zu können, wurden in den grundwassergespeisten Flüssen Main und Mosel sowie im Rhein Entnahmestellen oberhalb und unterhalb von Staustufen beprobt, die durch kontrastierende Strömungsverhältnisse als Modell für die zu erwartenden Änderungen dienten. Als Untersuchungsobjekt wurden Dreissena polymorpha PALLAS 1771 sowie Dikerogammarus villosus SOWINSKI 1894 herangezogen. Sie zeichnen sich durch hohe Abundanzen aus, sind aber unterschiedlich u.a. hinsichtlich ihrer Besiedlungsstrategie und –historie. Bei beiden Spezies sind die phylogeographischen Hintergründe bekannt; daher wurde auch versucht, die Einwanderungsrouten in der Populationsstruktur nachzuweisen (phylogeographisches Szenario). Dies konkurrierte mit der möglichen Anpassung der Spezies an das Abflussregime des jeweiligen Flusses (Adaptations-Szenario). Die Populationen wurden molekulargenetisch mit Hilfe der AFLP-Methode („Amplified-Fragment Length Polymorphism“) untersucht. Die Ergebnisse zeigen, dass D. polymorpha deutlich durch die Abflussregimes der Flüsse (Schmelz- oder Grundwasserdominanz) beeinflusst wird. Die Allelfrequenzen in Populationen des Rheins sind von denen der beiden grundwassergespeisten Flüsse Main und Mosel deutlich unterscheidbar (Adaptations-Szenario). Jedoch ist kein Unterschied der genetischen Diversitäten zu beobachten; das ist auf die lange Adaptation an ihre jeweiligen Habitate durch die lange Besiedlungsdauer zurückzuführen. Dies ist auch der Grund, warum die Einwanderungsrouten anhand der Populationsstruktur nicht mehr nachzuweisen waren. Die kontrastierenden Strömungsverhältnisse um die Staustufen hatten ebenfalls keine konsistenten Auswirkungen auf die genetische Diversität der Populationen. Diese Ergebnisse zeigen eine hohe phänotypische Plastizität der Spezies und dadurch eine große Anpassungsfähigkeit an wechselnde Umweltbedingungen, die unter anderem für den großen Erfolg dieser Spezies verantwortlich ist. D. villosus wanderte erst vor Kurzem in das Untersuchungsgebiet ein; die Einwanderungsroute war anhand der genetischen Diversität nachvollziehbar (phylogeographisches Szenario); durch die kurze Besiedlungsdauer war eine Adaptation an die divergenten Abflussregime der Flüsse nicht zu erwarten und wurde auch nicht gefunden. Dagegen war ein deutlicher negativer Einfluss von starker Strömung auf die genetische Diversität nachweisbar. Die Ergebnisse weisen darauf hin, dass die zukünftigen Auswirkungen des Klimawandels auf die Strömungsgeschwindigkeit negative Konsequenzen auf die genetische Diversität von D. villosus haben werden, während D. polymorpha hier keine Auswirkungen erkennen lässt. Die Auswirkungen des veränderten Abflussregimes im Rhein sind für D. villosus mit den vorliegenden Daten aufgrund der kurzen Besiedlungsdauer nicht vorhersagbar; D. polymorpha wird durch die Veränderung des Rheins zu einem grundwassergespeisten Fluss zwar einen Wandel in der genetischen Struktur erfahren, aber auch hier keine Einbußen in der genetischen Diversität erleiden.

Distinct lake level lowstand in Lake Prespa (SE Europe) at the time of the 74 (75) ka Toba eruption

Abstract. The 74 (75) ka Toba eruption in Sumatra, Indonesia, is considered to be one of the largest volcanic events during the Quaternary. Tephra from the Toba eruption has been found in many terrestrial and marine sedimentary deposits, and acidity peaks related to the eruption have been used to synchronize ice core records from Greenland and Antarctica. Seismic profiles and sedimentological data from Lake Prespa on the Balkan Peninsula, SE Europe, indicate a lake level lowstand at 73.6±7.7 ka based on ESR dating of shells. Tephrostratigraphy, radiocarbon dating and tuning of the total organic carbon content with the NGRIP isotope record, corroborate that the lake level lowstand was a short-term event superimposed on the general cooling trend at the end of MIS 5, most likely at the onset of the Greenland Stadial GS- 20. Acknowledging that tectonic events or karst processes could have triggered this lake level lowstand, the chronological correspondence between the lowstand and the Toba eruption is intriguing. Therefore a Toba-driven short-term shift to aridity in the Balkan region, leading to lake level changes and triggering spatial expansion events in one of the lake’s most abundant benthic species, the carino mussel Dreissena presbensis, cannot be excluded.

The Global Plankton Database: an inventory and data from the Former Soviet Union expeditions

At present time, there is a lack of knowledge on the interannual climate-related variability of zooplankton communities of the tropical Atlantic, central Mediterranean Sea, Caspian Sea, and Aral Sea, due to the absence of appropriate databases. In the mid latitudes, the North Atlantic Oscillation (NAO) is the dominant mode of atmospheric fluctuations over eastern North America, the northern Atlantic Ocean and Europe. Therefore, one of the issues that need to be addressed through data synthesis is the evaluation of interannual patterns in species abundance and species diversity over these regions in regard to the NAO. The database has been used to investigate the ecological role of the NAO in interannual variations of mesozooplankton abundance and biomass along the zonal array of the NAO influence. Basic approach to the proposed research involved: (1) development of co-operation between experts and data holders in Ukraine, Russia, Kazakhstan, Azerbaijan, UK, and USA to rescue and compile the oceanographic data sets and release them on CD-ROM, (2) organization and compilation of a database based on FSU cruises to the above regions, (3) analysis of the basin-scale interannual variability of the zooplankton species abundance, biomass, and species diversity.