570 Biowissenschaften; Biologie
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Wild boars belong to the most wide spread ungulates in the world. They are characterized by a well performed adaption to their environment mainly due to their omnivorous dietary. The wild boar population in Germany increased during the past three decades. Nowadays their high density leads to problems in agricultural areas due to damage of crops and plays a significant role as disease vector as the classical swine fever. For an effective population management population size information is of crucial importance. Different traditional methods exist to estimate population sizes as direct sightnings, faecal drop counts or hunting harvest which provide only relative estimates and population trends. Absolute population sizes could be yielded by a Capture-Mark-Recapture (CMR) approach. However, capturing of wild boars is difficult to realize and costly in terms of personnel and field effort.
Furthermore the capture probabilities are heterogeneous due to the variable behaviour of individuals influenced by age, sex, and experience of the animals. Non-invasive genetic methods are a promising complement to the traditional methods for population size estimation particularly for wild boar. These methods reduce stress and capture bias and increase the number of re-captures. Faeces proved to be a suitable DNA source for wild boar genotyping, due to almost equal capture probability. However working with faeces implicates difficulties such as low DNA rnquality and quantity, genotyping errors as dropout and false alleles.
The main aim of the present study was to develop a reliable, cost-efficient, reproducible and practicable method for wild boar genotyping. This method should provide a reliable dataset of genotypes obtained from the collected faeces samples. Individual identification forms the basis for an improved mark-recapture approach. As there is no sound method for absolute population counts in free living wild boar, reference values for the validation of this new approach are missing. Therefore, different routines to reduce and to assess genotyping errors were compared within this thesis. For maximum amplification rate, the storage, the extraction methods and the PCR-procedure were optimised. A step by step procedure was evaluated in order to determine the minimum required microsatellite (MS) number for reliable individual identification including a test with family groups (female and embryo tissue) to distinguish even between close relatives. A multiple-tubes approach, post-amplification checking and different correction procedures were applied to reduce genotyping errors. In order to quantify real genotyping error rates (GER) of datasets derived from sampling in the Palatinate Forest in western Germany, different methods for GER determination were compared with each other, obtaining GERs between 0% and 57.5%. As a consequence, more strict criteria for the multi-tube approach and increased repetition number of homozygous samples were used. An additional method validation was the implementation of a blind test to achieve the reliability of the genotyping and error checking procedure. Finally a strict and practicable proposal for the lab procedure was developed, by beginning with faecal sample collection and ending with a reliable dataset with genotypes of each sample.
The results of the presented method were derived from two sampling periods in a 4000 ha area in the Palatinate Forest in Rhineland-Palatinate in December 2006 and 2007. Both provided high confidence intervals (CI) applying inaccurate estimates (eg. for 2006 population size amounted to 215 with CI 95% of 156-314 and for 2007 population size amounted to 415 with CI 95% of 318-561) due to low sampling sizes (for 2006 n = 141 and for 2007 n = 326), successfully analysed samples (for 2006 n = 89 and for 2007 n = 156) and recapture numbers (for 2006 n = 12 and for 2007 n = 24). Furthermore, the population estimates even for the lowest values were considerably higher than previously assumed by hunting statistics, which implicates an ineffective hunting regime in the study area. For the future prospect, to obtain more precise population size estimations the increase of sampling sizes is inevitable, because absolute and reliable estimates are highly desirable for wildlife management and the control of diseases transmission. Nevertheless, the method for individual genotyping of wild boars evaluated in this thesis could be successfully established resulting in reliable datasets for population estimation modelling with sufficiently low GER.
Population genetic structure in European Hyalodaphnia species: Monopolization versus gene flow
(2012)
Cyclic parthenogens displays an alternation of asexual and sexual reproduction which has consequences for the genetic structure of these organisms. The clonal diversity of cyclic parthenogenetic zooplankton populations is influenced by the size of the dormant egg bank, i.e., the amount of sexually produced dormant eggs that assembled in the sediment, as these dormant eggs contribute new genetic variants to the populations. Further, the clonal diversity is impacted by clonal erosion over time, which reduces the number of different clones through stochastic and selective processes. Although freshwater invertebrates are good dispersers through their dormant stages, the influence of gene flow is assumed to be negligible, as the local population successfully monopolizes the available resources. As these populations reach carrying capacity fast due to the asexual reproduction, the first colonizing individuals are able to successfully establish in the habitat, resulting in a priority effect which hinders the invasion of new genotypes. Due to clonal selection and sexual reproduction a population will locally adapt over time and will establish a dormant egg bank which facilitates the fast re-colonization after a hostile period. This thesis evaluates the processes altering the population genetic structure of cyclic parthenogenetic zooplankton with a special focus on the concepts of monopolization as well as the counteracting effects of gene flow, using large-lake Daphnia species. Thirty-two variable microsatellite DNA markers were developed and a subset of twelve markers was evaluated regarding their suitability for species assignment and hybrid class detection. With this marker set and an additional mitochondrial DNA marker forty-four natural European populations of the species D. cucullata, D. galeata and D. longispina were studied. In D. galeata, most populations were characterized by low clonal diversities which suggest high influence from clonal erosion over the growing season and a low contribution from the dormant egg bank. Further, recent expansions as well as gene flow were detected, probably caused by the anthropogenic alteration of freshwater habitats, in particular eutrophication of many European lakes. D. longispina and D. cucullata revealed a different genetic structure compared to D. galeata, with high genetic differentiation among populations. This indicates low levels of effective gene flow which is in line with the predictions of monopolization. Further, high clonal diversities were found in populations of the two taxa, suggesting a high contribution from the dormant egg bank while clonal erosion was often not detectable. In D. longispina, mitochondrial data revealed an ancient expansion which was probably initiated by the formation of glacial lakes after the last ice age.
In addition, in D. longispina not only clonal diversity but also genetic diversity was high, indicating that during the build-up of the studied populations the influence from gene flow was probably high. To better understand the processes that act on early populations the population build-up in regard to the temporal advantage of clones during invasion succession was experimentally studied and revealed that priority effects shape population structure of Daphnia species. However, in certain cases the highly superior clones resulted in the extinction of inferior clones independent of the temporal advantage the single clones had.
This clearly shows that not only the time of succession is important but also the competitive strength. rnIn conclusion, the results obtained show that the population genetic structure in cyclic parthenogenetic zooplankton species is impacted by various processes. In addition to earlier studies, which mainly focus on local adaptation, clonal erosion and the size of the dormant egg bank to understand population genetic structure, this thesis could show that gene flow may be effective as well. During population build-up the advantage of early arriving individuals does not necessarily predict the outcome of population assembly, as additional genotypes may contribute to the population. Finally, the genetic structure of established populations may be severely impacted by effective gene flow, if severe environmental changes alter the habitat of the locally adapted population.
To assess the effect of organic compounds on the aquatic environment, organisms are typically exposed to toxicant solutions and the adverse effects observed are linked to the concentration in the surrounding media. As compounds generally need to be taken up into the organism and distributed to the respective target sites for the induction of effects, the internal exposure is postulated to best represent the observed effects.
The aim of this work is to contribute to an improved effect assessment of organic compounds by describing experimental and modelling methods to obtain information on the internal exposure of contaminants in organisms.
Chapter 2 details a protocol for the determination of bioconcentration parameter for uptake (k1) and elimination (k2) of organic compounds in zebrafish (Danio rerio) eggs. This enables the simulation of the internal exposure in zebrafish eggs from an ambient exposure concentration over time. The accumulated contaminant amount in zebrafish eggs was also determined, using a biomimetic extraction method. Different bioconc-entration estimation models for the determination of internal steady-state concentrat-ion of pharmaceutical compounds in fish to an environmental exposure are presented in Chapter 3. Bioconcentration factors were estimated from the compounds octanol: water partition coefficient (KOW) to determine the internal exposure to an ambient concentration.
To assess the integral bioavailable fraction from the water and sediment phase of environmental contaminants for rooted aquatic plants, the internal exposure in river-living Myriophyllum aquaticum plants were determined over time, presented in Chapter 4. The plants were collected at different time points, with the accumulated organic contaminants determined using a liquid extraction method.
In Chapter 5 a protocol was established to enable the non-invasive observation of effects in M. aquaticum plants exposed to contaminated sediments over time. Since the toxicant effects are a result of all uptake and distribution processes to the target site and the toxico-dynamic process leading to an observed effect during static exposure, information on the internal exposure could thus be gained from the temporal effect expression.rn
Nandi forests (South and North Nandi forests) are situated in the Rift Valley Province of Kenya very close to Kakamega forest. From previous documents it has been seen that Kakamega and Nandi forests were connected to each other forming one big "U" shaped forest block till the beginnings of 1900s. Due to human pressures, currently there are three different forests form the previous one block forest. Although they were one forest, information on Nandi forests is very scanty when it is compared to that of Kakamega forest. The species composition and diversity as well as plant communities and population structure of Nandi forests have not been studied. Information is not available about the similarity status of South and North Nandi forests. Furthermore the natural regeneration potential (seedling bank) of these forests is not well studied and documented. Hence this study aims to fill these gaps.
In this study totally 76 quadrates (49 from South Nandi and 27 from North Nandi) were used to collect data. In the South Nandi forests 27 of the quadrates were laid in the better side of the forest (at Kobujoi) and the remaining 22 were in the heavily disturbed part of this forest (Bonjoge). The quadrates were arranged on transects that have one to one and half km which were parallel to the slope. The distance between the quadrates was 100 meter and transects are 500 m apart. The size of the main quadrate was 400 m2 (20 X 20 m) which also had five small plots (3 X 3 m) distributed on the four corners and in the center. Each woody plants (climbers, shrubs and trees) having more than one meter and greater than two centimeter diameter at breast height (dbh) were measured and recorded. Seedlings and herbaceous plants were sampled in the smaller plots. Individual plants were identified at species level and when it was not possible to identify in the field voucher specimen were prepared and latter identified at the East African Herbarium, National Museum of Kenya, and Nairobi. Clustering and ordination were performed using PC-ORD and CANOCO ecological softwares, respectively. For both clustering and ordination abundance data of the species was used. Shannon diversity index and evenness were computed using PC-ORD while similarity indices, Fisher alpha, rarefaction, species richness estimation (nonparametric species richness estimators) were conducted using EstimateS. Indicator species analysis was undertaken using PC-ORD. Basal area and height class distribution at forests level or site level (Bonjoge and Kobujoi) and diameter (dbh) class distribution for selected trees species were performed to evaluate population structure.
Furthermore importance value (IV) of woody plant species was calculated. SPSS version 16 was used to undertake both parametric (when data assume normal distribution) and nonparametric (when data are not assuming normal distribution) comparison of means, correlation and regression analysis.
In this study totally 321 vascular plant species comprising 92 families and 243 genera were identified in Nandi forests (both South and North Nandi forests). In South Nandi forest 253 plant species form 82 families and 201 genera were recorded while in North Nandi 181 species comprising 67 families and 155 genera were recorded. Jackknife second order estimators gave the highest species richness estimate for both South and North Nandi forests i.e. 284 and 209, respectively. In the case of highly disturbed and less disturbed parts of South Nandi forest 138 and 172 vascular plant species were recorded, respectively. Asteraceae, Rubiaceae and Euphorbiaceae are the top three species rich families of Nandi forests. In terms of different diversity measures (i.e. alpha and beta diversity, Fisher alpha, Shannon diversity and evenness indices) South Nandi is more diverse than North Nandi forest. Sörensen and Jaccard (classic) as well as their respective abundance based similarities showed that there is a low species similarity between South and Nandi forests. The cluster analysis resulted in three different plant communities and this result is supported by the ordination result.
South and North Nandi forest has inverted "J" height class distribution showing that larger proportion of woody plant individuals are found in the lower height classes. Similar pattern is observed when the diameters of all woody plants were considered together. However, different diameter class distributions (seven types) were identified when selected tree species were analyzed separately. It has been observed that the basal area of South Nandi forest is significantly lower than that of North Nandi forest (Mann-Whitney U =358, p < 0.001). Similarly Bonjoge has significantly lower basal area (t-value=3.77, p<0.01) than that of Kobujoi. Number of woody plat seedlings in South Nandi forest is significantly higher than that of North Nandi (Mann-Whitney U = 362.5, p<0.001). In the same way Bonjoge has significantly smaller number of ssedlings than Kobujoi (t-value 4.24, p<0.001). Most of species in both forests are able to resprout from stumps after physical damage; hence this helps the regeneration of the forests in addition to seedling banks. This study enables to fill some of the information gaps about Nandi forests especially of floristic composition, population structure, natural regeneration and human impacts on this ecosystem.