5. RESULTS
5.7 Body burdens
Body burdens are important for evaluation of accumulation and transfer of fullerene in aquatic food chains. Organisms with bigger body size can ingest more. Biouptake may is the main option for fullerenes route upward in aquatic food chains. It was not possible to extract fullerenes from the samples because of too low biomass for extraction. This situation was different from stirred suspension with larger particle sample because tiny particles don’t stick to the gut wall as strongly as bigger ones.
33 6. DISCUSSION
6.1 Findings of suitable concentration for low toxicity TPT on Daphnia magna
In the present study show that most of the D. magna were immobilized at 300µg/l treatment. Six concentrations were used for measurement suitable concentration for chronic tests, 0.1 µg/l all daphnia were mobile at 24h and at 48h up to 80% whereas in control 90% was mobile. In the test 50% immobility was not reach in any treatment group which indicated only low acute toxicity of fullerenes on D. magna as noted also in previous studies (e.g. Henry et al. 2011).
Percentage of immobility depends on water stability that in turn depends on the agglomerate size of .
6.2 The exposure affects the body size of F1 and F2 generation.
Exposure of F0 generation affected the body size of F1 and F2 generations. In F1 generation had bigger body size compared to the control group (Figure 24 A). Each generation body size also higher compares to each control group except F2 generation (Figure 24 B). But F1 generation mother sample smaller compare to F2 generation mother samples. Carbon nanomaterial’s have shown to induce changes in the size of adult D. magna but also in the number of neonates.
Increased size was determined for each exposure group. Due to environmental stress, life strategy there is a shift to produce fewer neonates (reduced number of offspring) of larger neonates size (Arndt 2014).
Changed body sizes of next-generation can be a result of variation in factors such as temperature, food habit, environmental stress, genetic and maternal (Heugens et al., 2006, Ebert 1993). These previous findings are in accordance with those made in this study., F1 generation body sizes were increased and F2 generation body sizes were increased. Decrease of neonate body lenght depends also on mother’s age (Ebert 1993).
Previously it has been shown that, there is positive relationship EC50 (24 and 48h) and neonate body size, a smaller size of Daphnia species the higher the sensitivity to heavy metal toxicity (Vesela et al. 2006). This effect is a species-specific. It has also been shown that larger species of Daphnia living to permanent water, where the species have a much smaller body (Hebert 1978). In this study 200ml jar/8 daphnia then transfer 50ml jar/1 daphnia for 28 days. I
34
think here is main factor to environment for larger bodies of daphnia and produce of fewer neonates. When larger species were living small habitats has been most convincingly predation by fish. Body sizes were the direct relationship of food particles ingest (Hebert 1978).
6.3 Sex ratio
The results showed that almost all individuals were females in different treatments and the production of males would indicates poor conditions. Sex of invertebrates is determined by temperature, daylight, nutrition, density, humidity, pH, ionic composition UV light, metabolic products, host size, age, and type, etc (Heugens et al. 2006). In addition, parasites can affect the determination of sex in invertebrates (Korpelainen 1990). Changes in food levels does not affect the determination of sex if the starvation is not strong. In crowed ponds 43% can be made as a response to the metabolites of daphnias. A very short day length results also production of male offspring (Hobaek and Larsson 1990).
Artificial fresh water and 16:8 h photoperiod was used in this study. The population density was not especially high around 200ml for 8 daphnias and this followed guidliens (OECD/OCDE). So results showed that, F1 and F2 generation treatment group, 99.20% female and also tin group 100% female (Table 4).
6.4 Nanoparticle affect the number of neonates, birth rate, and mortality rate in Daphnia magna in studied generations
The number of neonates was high in the F1 generation in all treatment groups compared to control group were high in the treatment group compared to control (Figure 26A). Results also showed that in F2 generation the number of neonates was high in all treatment group compare to control group. The body size also high in treatment compared to control (Figure 26B).The number of neonates depend on body size. Increase in body size results in increased number of neonates. For example the exposure of Daphnia magna species to zinc has also induced larger bodies of higher number of neonates produced (Vessel et al. 2007). In this study fullerene and orgonotin compounds were shown to be toxic to D. magna but larger bodies with fewer neonates was observed. On the other hand smaller size of Daphnia, means higher sensitivity to heavy metal toxicity (Vessel et al. 2007).
35
Additionally it has been shown that, exposure of filtered fullerenes and filtered TiO2 increases mortality and increase higher level of toxicity. Sonication of fullerene solution increases the mortality rate (Lovern and klaper 2006).
Mortality was higher in F0 generation compared to the control group. Mortality was also high in F1 mother generation in Tin treatment. Lovern and later (2006) have shown that,sonication increases mortality 9% of D. magna in 500mg/L nTiO2 suspension. However, in this study mortality rate was very low. The toxicity of nanoparticles depends on particles size, preparation method, and also test design. In most studies sonication in creases the toxicity of fullerenes (Zhu et al. 2009).
7. CONCLUSION
Nanoparticles has been studied for acute and chronic ecotoxicological effects on freshwater zooplankton like Daphnia species. The present work clearly shows that fullerene and orgonotin compounds are potentially more toxic to D. magna upon joint exposure. In addition, fullerene and organotin change the life cycle of Daphnia also by reducing the number of offspring.
Fullerenes are very stable molecules. This stability allows to potentially accumulate in aquatic environment which may increase the environmental concentration of fullerene. The environmentl fate of fullerens depend on the characters of water.
From the observations made in the experiments presented in this thesis, are ingested by Daphnia magna. This effective intake makes the problem for other species in a tropic level transferring system. This experiment also observed that very low concentrations (0.1µg/l) of Triphenyl tin may cause toxic response occurred in the Daphnia both in chronic and jointly with fullerene in F1&F2 generation. For example there was changes in growth rate, body size, reproduction, and mortality rate Joints effects follow, so if one generation is exposed be affected the next generations ill be affected.
This study also indicates the ability to excrete fullerene from the gut tract on daphnia species in higher body weight, but also found a very few amount in small weight daphnia sample.The tested extraction method was not sensitive enough to quantify fullerne in Daphnia, because of small sample size. To concluded behavior of fullerenes in the environment has been studied insufficiently and requires close attention because a release of nanomaterial may be huge in the near future.
36 ACKNOWLEDGEMENTS
The present study was conducted at the Department of Environmental and Biological Sciences of the University of Eastern Finland (Joensuu Campus). I am very grateful to this Institute for a great opportunity to do my Master’s Thesis. I would like to thank Dr. Jarkko Akkanen and Dr.
Kukka Pakarinen for supervision and helpful advice during my practical work and the writing process. My warmest thanks to my supervisor for valuable comments I got on manuscript, helped me a lot to improve my thesis. Furthermore, I thank the rest of the staff of the aquatic Ecotoxicology group in the University of Eastern Finland (Joensuu).
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