Effects of ozonation treatment on benthic fauna
How does the density of aquatic invertebrates change in the river Knivsta during removal of pharmaceuticals?
Ludvig Hagberg
Student
Degree Thesis in Biology 15 ECTS Bachelor’s Level
Report passed: XX June 2016 Supervisor: Tomas Brodin
Effects of ozonation treatment on benthic fauna – How does the density of aquatic invertebrates change in the river Knivsta during removal of
pharmaceuticals?
Ludvig Hagberg
Abstract
The quantity of pharmaceuticals that end up in aquatic and terrestrial ecosystems are
increasing due to a global increase in usage and poor removal efficiency of pharmaceuticals in sewage treatment plants (STP). These compounds may cause great effects on both aquatic and terrestrial ecosystems. Pharmaceuticals are biologically active compounds that may affect non-target organisms. This report focused primarily on if and how the density of Chironomidae (nonbiting midges), Isopoda, Trichoptera (caddisfly) and Simuliidae (black fly) changed when sewage water was treated with ozonation in the river Knivsta, south of Uppsala. Invertebrates were sampled using the kick-sampling method at 6 sites downstream a STP both before and during ozonation treatment. Furthermore, the biodiversity of
invertebrates before and after treatment was studied. Significant differences between densities of Isopoda and Gastropoda were found when comparing samples collected before and during ozonation. Both groups were more abundant during ozonation treatment than before. No trend could be found for effects on biodiversity. Further studies are needed to determine which effects pharmaceuticals have on non-target organisms and also if
pharmaceutical removal techniques such as ozonation may have other effects, not related to the removal of pharmaceuticals, on ecosystems and the wildlife therein.
Table of contents
1. Introduction 1
1.1 Background 1
1.2 Changes in behavior due to pharmaceuticals 2
1.3 Effects on benthic fauna 2
1.4 Problem statements 3
2. Method and materials 3
2.1 Method 3
2.2 Statistical analysis 4
3. Result 5
3.1 Biodiversity 5
3.2 Invertebrate density 5
4. Discussion 7
4.1 Biodiversity 7
4.2 Invertebrate density 8
5. References 10
1
1. Introduction
1.1 Background
Pharmaceuticals are used globally on a daily basis, and in the last decade more and more pharmaceutical residues have been found in aquatic systems (Daughton and Ternes 1999).
This is mainly due to a poor removal efficiency of pharmaceuticals in our sewage treatment plants (STP) (Brodin et al. 2014, Grabicova et al. 2015 and Jonsson et al. 2015). The
concentrations of pharmaceuticals in aquatic systems are usually low and therefore hard to detect. Only recently with improved analytical technologies have those compounds been observed in the environment, usually in the ng–μg/L range. The active substances in pharmaceuticals may have a great impact on different ecosystems, both aquatic and
terrestrial (Sánchez-Argüello, Fernàndez and Tarazona 2009, Gilroy et al. 2012, Arnold et al.
2013 and Brodin et al. 2014).
In total over a hundred different pharmaceutical compounds have been found in the environment and presumably there are many more to be discovered (Guo et al. 2016 and Sumpter 2016). There are more than 4000 pharmaceuticals currently on the market, including antibiotics, antihistamines, analgesics and hormones, both for human and veterinary uses. Those compounds as well as personal care products are biologically active compounds which interact with specific processes in the target organism, but may also pose a threat for non-target organisms, in the environment, that carry similar receptors (Marshall and Williams et al. 2011, Royer 2012, Boxall et al. 2012 and Klaminder et al. 2015). As much as 90% of the active drug substance can still remain active after leaving the target organism (Guler and Ford 2010). The degradation rate, toxicity and the rate at which those
pharmaceuticals enter non-target organisms can vary depending on the pH of the environment (Williams et al. 2011). Temperature is also an important factor for the degradation rate of pharmaceuticals. The degradation rate increases with warmer
temperature and with an increase in pH (Webster, Mackay and Wania 1998). For lindane, an insecticide used in both agriculture and as a treatment for lice and scabies, the half-live can vary from 75 days at 30◦C up to one hundred years at temperatures close to zero (Webster, Mackay and Wania 1998). Even bigger changes in degradation rate can be found when looking at different pH levels. At pH 3 the half-live of lindane is as long as 175 000 years while at pH levels around 9 only 64 days (Webster, Mackay and Wania 1998). Biological factors like habitat preference, life cycle stage, and reproductive strategies may also affect an organism’s uptake of pharmaceuticals (Williams et al. 2011). Organisms that lives on the bottom of a lake or stream may have a greater uptake of pharmaceuticals from the soil compared to organisms that lives exclusively in vegetation (Rubach et al. 2011). However, a predator which feeds on contaminated prey may be more exposed to pharmaceuticals through food intake in comparison to habitat preference (Rubach et al. 2011).
Ozone is a gas which is very unstable. The rate of degradation in water ranges from a few seconds to 30 minutes (Water research center 2014). The molecule is composed of three oxygen atoms and due to its instability it’s degrading to O2 which is a far more stable state.
Once this happens there is one free oxygen atom which is highly reactive. The free oxygen atom can oxide iron and other metals in water and turn them into an insoluble metal that are removed by filtration. Ozone can also eliminate organic particles, chemicals and reduce odor in water (Water research center 2014). The ozone also has to be removed from the water because of its reactive properties; this can be done using a helium stream (Rodríguez et al.
2013). Rodríguez et al. (2013) has shown that pharmaceuticals from water may be removed in under 30 minutes with the ozonation technique.
According to Jiang and Zhou (2013), ferrate (FeVIO42−, Fe (VI)) is a chemical which can remove more than 80% of ciprofloxacin in waste water effluent. It may also remove other
2
pharmaceuticals such as ibuprofen and naproxen. The use of ferrate with its dual function as an oxidant and coagulant is therefore a promising technique for removal of pharmaceuticals.
This technique is however pH dependent and works best at pH levels around 4 (Jiang and Zhou 2013).
1.2 Changes in behavior due to pharmaceuticals
Studies have shown significant behavioral alterations in fish due to pharmaceutical impact.
For example, European perch (Perca fluviatilis) become more active and bolder and at the same time less social in the presence of the anxiolytic pharmaceutical oxazepam (Brodin et al.
2013 and Brodin et al. 2014). Another study showed that when hybrid striped bass (Morone saxatillis x Morone chrysops) got exposed to sub-lethal concentrations of fluoxetine, an anti- depressant, their ability to catch prey decreased which can lead to reduced fitness (Gaworecki and Klaine 2008). Gaworecki and Klaine (2008) also suggested that serotonin may be used as a biomarker due to its ability to remain in the organism under a non-exposure period as well. Invertebrates are also affected by pharmaceuticals and other pollutants. Fluoxetine in the environment can alter endocrine functions and serotonin levels among invertebrates (Guler and Ford 2010). Changes in serotonin levels among invertebrates may have many effects like changes in reproduction, swimming behavior, egg laying and feeding behavior (Guler and Ford 2010). De Lange et al. (2009) showed that Gammarus pulex exposed to fluoxetine, carbamazepine or ibuprofen showed a dual response for each pharmaceutical. At low concentrations (1-100ng/L) an increase in ventilation (resting) could be found and higher concentrations (1µg/L–1 mg/L) resulted in increased locomotion. A component called CTAB that are used in antiseptics had a more acute effect on G. pulex. A correlation could be found when comparing CTAB concentration with inactivity for G. pulex. At doses between 10- 100mg most individuals died within 2 hours (De Lange et al. 2006). Some pharmaceuticals have a more direct impact on species at even lower concentrations. For example in Pakistan the oriental white-backed vulture nearly got extinct because of high levels of diclofenac in livestock that the vultures feed on (Oaks et al. 2004). Other effects from pharmaceuticals range from changes in insect physiology and behavior to effects on algae or aquatic plant growth (Williams et al. 2011).
1.3 Effects on benthic fauna
Studies have shown that sediments in aquatic systems may act as a long term source of pharmaceutical compounds due to higher concentrations of those compounds in the
sediment compared to the surrounding water (Gilroy et al. 2012). Benthic organisms exposed to such sediment could then be affected (Gilroy et al. 2012). Due to low concentrations of pharmaceuticals in STP effluent water these compounds have gone undetected in aquatic systems until the last 15 years. Therefore, the knowledge of which ecological effects
pharmaceuticals may have in aquatic ecosystems are very limited, however more and more studies are focusing on this subject today. More studies on this subject are needed to get a better understanding of the short- respectively long-term effects of pharmaceuticals in our environment.
This study focuses on if and how biodiversity and density of benthic fauna are affected by pharmaceuticals in water. More specifically it tests if the density of benthic fauna changes when pharmaceutical residues are removed, by ozonation, from treated sewage water entering a river.
3
1.4 Problem statements
1. How does the biodiversity of aquatic insects change when sewage water is treated with ozonation before entering the river?
2. How does the density of Chironomidae (nonbiting midges), Isopoda, Trichoptera (caddisfly) and Simuliidae (black fly) in the river Knivsta change when sewage water is treated with ozonation before entering the river?
3. Are there any aquatic insects that can only be found in the treated respectively untreated water?
2. Method and materials
2.1 Method
Knivsta sewage treatment plant (STP) was renovated during 2013/2014 to be able to clean pharmaceuticals from sewage water. In June 2015 this STP was the first full scale STP with ozonation in Sweden. Studies have shown that pharmaceutical levels will be reduced between 80 and 99.9% (Jerker Fick personal commentary 2016). Benthic organisms included in this study were obtained from the river Knivsta, Knivsta, located south of the city Uppsala.
Invertebrate samples were collected from six sites (5-10) shown in figure 1 using the kick- sampling method with five subsamples in each site per sampling event (Appendix 1). Kick- sampling means that a net is “kicked” along the riverbed for a specific distance (here 50 cm) and if the ground is to muddy, gently dragged along the ground. The content collected using the kick-sampling method were put in plastic containers and marked with site number and date. All of these sites were located downstream from the sewage treatment plant. 30 samples were collected in spring (April-May) 2015, before the ozonation treatment began, and 30 in the autumn (October-November) 2015 while the ozonation treatment was in progress.
Figure 1. Map of river Knivsta with sample locations marked. The red square indicates where the sewage treatment plant is located with samples numbered from 5-10 placed downstream from the STP.
4
After the sample collection from the river, the content of each container were analyzed at Umeå University using a stereo microscope. All invertebrates found in each container were put in different small glass vials depending on which taxon they belonged to. For example, all chironomids from the same sample were put in the same vial. A 70% ethanol solution was added to every glass vial to conserve the invertebrates. Furthermore, all invertebrates were counted and assigned to categories of each taxa, sample and date.
2.2 Statistical analysis
All statistical analysis were made in Microsoft Excel with a 95% confidence interval for significance. If the data was normally distributed, T-tests and one way ANOVA’s were used to investigate changes between species composition before and during the ozonation treatment.
When data were not normally distributed, the non-parametric Kruskal–Wallis one-way analysis of variance were used. Firstly all data had to be ranked from 1-60. All tied values with the same number had to be averaged to get the rank they would have received had they not been tied. N is the total number of observations across all groups. ni is the number of observations in group i and the ri is the average rank of all observations in group I.
Figure 2. Calculations for the Kruskal-Wallis one-way analysis.
To calculate the diversity index in all the samples the Shannon-Wiener diversity index were used.
Figure 3. Calculations for the Shannon-Wiener diversity index.
Thereafter a one way ANOVA could be used to calculate the differences in diversity before and during the ozonation treatment.
5
3. Results
3.1 Biodiversity
There was a higher biodiversity at site 10 before ozonation treatment (1.79 ± 0.07) compared to during the treatment (1.31 ± 0.17) and this difference was significant (F = 21.86, p = 0.002). At site 7 the opposite pattern was found with a higher biodiversity during the treatment (1.34 ± 0.03) than before (1.12 ± 0.09) and this was marginally significant (F = 5.12, p = 0.054; fig 4). No other significant results could be found when analyzing
biodiversity before and after removal of pharmaceuticals.
Figure 4. Shannon-Wiener diversity index for site 7 and 10 both before and after of removal of pharmaceuticals in sewage water. Error bars denote ± 1 standard error (SE).
3.2 Invertebrate density
More Gastropoda were found during ozonation treatment (1.47 ± 0.32) compared to before the treatment (0.2 ± 0.09) when analyzing all sites collectively. This difference was
significant (p = <0.004; Fig 5). Other taxa showed no significant difference when all sites were compared together.
Figure 5. Mean values of Gastropoda sample both before and during ozonation treatment. Error bars denote ± 1 standard error (SE).
6
There were considerable more Gastropoda during ozonation treatment at three different sites compared to before the treatment. At site 6 no Gastropoda were found before ozonation treatment. The average number of Gastropoda per sample during ozonation treatment was 3
± 0.632 and this increase was significant (p= 0.003). Similarly, no Gastropoda were found before ozonation treatment at site 7 either. The average number of Gastropoda per sample during ozonation treatment was 3.4 ± 0.927 and this difference was also significant (p=
0.003). At site 10 an increase from 0.6 ± 0.4 to 2 ± 0.447 could be found. This difference was marginally significant (p = 0.06; Fig 6).
Figure 6. Mean values of Gastropoda sample both before and during ozonation treatment. Error bars denote ± 1 standard error (SE).
No significant difference could be found when comparing Isopoda for all sites collectively.
The same trend was found for Isopoda with more Isopoda found at both site 5 and 6 during ozonation treatment compared to before the treatment. The average number of Isopoda per sample at site 5 increased from 2 ± 1. 05to 9.2 ± 1.93 and this difference was significant (F = 10.71, p = 0.011). At site 6 the average number of Isopoda per sample was 1.4 ± 0.24 before removal of pharmaceuticals and increased to 5.4 ± 1.69 during the ozonation treatment. This difference was also shown to be significant (F = 5.48, p = 0.047; Fig 7).
7
Figure 7. Mean values of Isopoda/sample both before and during ozonation treatment. Error bars denote ± 1 standard error (SE).
No significant differences could be found when looking at Chironomidae, Trichoptera and Simuliidae per site, nor when comparing all sites collectively. Anisoptera (dragonflies) were only found before the ozonation treatment but only two individuals in total. Five taxa were only found in the ozonation treated water ranging from 1-6 individuals: 4 Tipulidea (crane flies), 3 Corixidae, 6 Lepidoptera (moths and butterflies), 1 Nepidae (water scorpions) and 1 Zygoptera (damselflies). In total 15 taxa were found before ozonation treatment and 19 taxa during the treatment (Appendix 2).
4. Discussion
4.1 Biodiversity
No clear patterns could be found when studying biodiversity in effluent water from a STP before and during ozonation treatment. When we compared site to site, a difference could be found at two different sites, 7 and 9. However, those two results differed in the direction of the effect as site 7 had a higher biodiversity during the treatment and site 9 before the treatment. This study suggests that biodiversity isn’t affected in a clear way by
pharmaceutical compounds in aquatic systems. Most likely those relatively low
concentrations (ng-µg/L) of pharmaceutical in the environment are not lethal and thus no difference in diversity could be found. Toxicology studies often use pharmaceuticals at concentrations around 1-300mg to test mortality of different organisms when exposed to pharmaceuticals (Nunes, Carvalho and Guilhermino 2005, Nalecz-Jawecki and Persoone 2005 and Coelho et al. 2011). If some species or groups of species are affected more than others they might slightly reduce population size but then a niche is available for other species to occupy which are not as affected by the pollution. This would not show when analyzing the diversity in an area and therefore more studies on how different taxa’s behaviors changes when exposed to pharmaceuticals are surely needed.
The sampling of data before and during ozonation treatment wasn’t collected at the same time of the year. One was collected in spring and the other in autumn, which might cause a temporally induced difference in the data. Some of the insects may live in the stream as
8
larvae and then leave when they are fully developed, if they undergo this change during summer or early autumn this may cause quite the difference in population size. However, between November and April, as were sample-times in this study, the production of biomass and change in biotic community in streams are very low due to low temperatures. Thus no significant changes due to temporal effects should occur during this time a year. Although if many species developed into their adult stage during summer and left the river, a lower number of species may be found in the autumn in comparison to what would have been found during spring. Ivkovic et al. (2013) states that an increase in water temperature and length of sunlight is two important factors when studying emergence of aquatic invertebrates.
Many aquatic insects tend to emerge from the river when water temperatures exceeds 16◦C. If the sampling of data would have been in spring 2 subsequent years a difference in
biodiversity might have been found. Mainly due to a longer ozonation treatment. It is unclear if 3 months of ozonation is enough to remove all pharmaceuticals from the water and the sediment. Thus a longer period of ozonation would have been preferred before the sampling began.
4.2 Invertebrate density
More Gastropoda during ozonation treatment than before could be found when comparing all sites downstream the STP. The same result was found in 3 of the 6 sites when comparing changes within sites. Gastropoda is clearly affected by, at least some, pharmaceuticals in their environment. Fluoxetine, an anti-depressant commonly known as Prozac and Sarafem is only one of many selective serotonin reuptake inhibitors (SSRIs) that are used widely today.
Serotonin exist in the nervous system among many vertebrates and invertebrates. SSRIs inhibits the reuptake of serotonin thus increases the serotonin neurotransmission which affects many physiological functions in mollusks like Gastropoda (De Lange et al. 2006). It also affects reflexes like heartbeat rhythm and behaviors like feeding, locomotion patterns and reproduction (De Lange et al. 2006 and Daughton and Ternes 1999). Fluoxetine may affect behavior at very low concentrations (100ng/L) which is observed in STP effluents (De Lange et al. 2006). Consequently if low concentrations of pharmaceuticals may have such a big effect on organisms in the environment the lack of Gastropoda before removal of pharmaceuticals in this study may be explained by the presence of pharmaceuticals in this aquatic system. When the ozonation treatment began and the water became less polluted, organisms that are sensitive to pharmaceutical and other pollutants might have migrated downstream or upstream to get closer to the cleaner water near the STP.
Freshwater Gastropoda may be used as biological indicators. Some radioactive materials and heavy metals are concentrated in their flesh and shell (Hart and Fuller 1974). Therefore, it would be interesting to study if pharmaceuticals also concentrate in their body like other pollutants. Gastropoda and other mollusks could be interesting subjects to study in the future and more studies would be necessary to determine if they may be used as indicators for pharmaceutical and other similar pollutants.
Significantly more Isopoda were found at 2 sites (5 and 6) during ozonation treatment in comparison to before. De Lange et al. (2009) showed that low concentrations (1-100ng/L) of ibuprofen, fluoxetine and carbamazepine had an effect on G. pulex. An increase in ventilation (resting) could be found. G. pulex are related to Isopoda on a class level, both belong to Malacostraca. Therefore there may be similar effects on Isopoda that might explain this result. An increase in resting can lead to lower food intake which in turn leads to lower fitness and less reproduction.
9
No significant result could be found when studying the number of individuals for many other species groups like Chironomidae (nonbiting midges), Trichoptera (caddisflies) and
Simuliidae (black flies). This does not mean that these taxa are unaffected by pharmaceuticals, but that the number of individuals in this study did not change
significantly. Other biological endpoints such as behavior might have been affected. Studies have shown that fish behavior can change due to pharmaceuticals in their environment. For example, European perch (Perca fluviatilis) were shown to become bolder, more active and less social in the presence of oxazepam (Brodin et al. 2013 and Brodin et al. 2014). This may also be the case for some invertebrates and thus no changes in quantity could be found. Even though no direct changes were found for many species groups pharmaceutical may bio accumulate in invertebrates who gets exposed to those compounds during a long time. Later if they get eaten by fish for example these compounds will affect fish’s as well. Some
pharmaceuticals which increase feeding rate for consumers might cause an even higher increase in bioaccumulation with a higher food intake (Brodin et al. 2014). Biomagnification in aquatic systems can therefore be an important factor to keep in mind when studying pharmaceuticals in aquatic systems. Due to bioaccumulation the level of pharmaceuticals in surface water does not directly have to represent the levels found in organisms (Brodin et al.
2014).
More taxa could be found during the ozonation treatment compared to before. Those taxa occurred only in small quantities ranging from 1-6 individuals per taxa. Therefore it is hard to know if those taxa are especially sensitive to pharmaceutical compounds or if these
differences are natural random occurrences. Studies with larger sampling sizes are hence needed to exclude such anomalies.
Lürling, Sargant and Roessink (2005) showed that Daphnia pulex, which is a freshwater grazer showed an increase in reproduction when exposed to the pharmaceutical
carbamazepine. They were also larger and matured and reproduced earlier. So this species showed an increase in individuals due to pharmaceuticals. These various responses that different organisms tend to have as an effect of pharmaceuticals make it hard to predict general responses of how organisms are affected when exposed to pharmaceuticals.
Ecosystems are diverse and complicated systems which makes it extremely hard to understand all the processes and interactions therein. Pharmaceuticals have been used during the last century and especially the last 50 years (Rowland et al. 2012). Therefore, ecosystems have had been exposed to those compounds for much longer than we have studied its effects. The removal of pharmaceutical with different removal techniques like ozonation may have a greater effect on ecosystems than expected. More studies are therefore needed to assess the effects which the removal of pharmaceuticals has on both different taxa and on the interactions between species in aquatic systems.
10
5. References
Arnold, K. E., Boxall, A. B. A., Brown, A, R., Cuthbert, R. J., Gaw, S., Hutchinson, T. H., Jobling, S., Madden, J. C., Metcalfe, C. D., Naidoo, V., Shore, R. F., Smits, J. E., Taggart, M. A, Thompson, H. M., 2013. Assessing the exposure risk and impacts of pharmaceuticals in the environment on individuals and ecosystems. Biology letters 9:
20130492.
Boxall, A. B. A., Rudd, M. A., Brooks, B. W., Caldwell, D. J., Choi, K., Hickmann, S., Innes, E., Ostapyk, K., Staveley, J. P., Verslycke T., et al. 2012. Pharmaceuticals and Personal Care Products in the Environment: What Are the Big Questions? Environmental Health Perspectives 120 (9): 1221-1229.
Brodin, T., Fick, J., Jonsson, M., Klaminder, J., 2013. Dilute concentrations of a psychiatric drug alter behavior of fish from natural populations. Science 339: 814-815.
Brodin, T., Piovano, S., Fick, J., Klaminder, J., Heynen, M., Jonsson, M., 2014. Ecological effects of pharmaceuticals in aquatic systems - impacts through behavioral alterations.
The Royal Society Publishing 369: 1-10.
Coelho, S., Oliveira, R., Pereira, S., Musso, C., Domingues, I., Bhujel, R. C., Soares, A. M. V.
M., Nogueira, A. J. A., 2011. Assessing lethal and sub-lethal effects of trichlorfon on different trophic levels. Aquatic Toxicology 103 (3-4): 191-198.
Daughton, C., Ternes, T., 1999. Pharmaceuticals and personal care products in the
environment: agents of subtle change? Environmental Health Perspectives 107 (6):
907-938.
De Lange, H. J., Noordoven, W., Murk, A. J., Lürling, M., Peeters, E. T. H. M., 2006.
Behavioural responses of Gammarus pulex (Crustacea Amphipoda) to low concentrations of pharmaceuticals. Aquatic Toxicology 78: 209-216.
De Lange, H. J., Peeters, E., Lurling, M., 2009. Changes in ventilation and locomotion of Gammarus pulex (Crustacea, Amphipoda) in response to low concentrations of pharmaceuticals. Human and Ecological Risk Assessment 15: 111–120.
Fick, J., 2016. E-mail 2016-04-27. <Jerker.fick@umu.se>
Gaworecki, K. M., Klaine, S. J., 2008. Behavioral and biochemical responses of hybrid striped bass during and after fluoxetine exposure. Aquatic Toxicology 88: 207–213
Gilroy, E. A. M., Balakrishnan, V. K., Solomon, K. R., Sverko, E., Sibley, P. K., 2012.
Behaviour of pharmaceuticals in spiked lake sediments – Effects and interactions with benthic invertebrates. Chemosphere 86 (6): 578-584.
Grabicova, K., Grabic, R., Blaha, M., Kumar, V., Cerveny, D., Fedorova, G., Randak, T., 2015.
Presence of pharmaceuticals in benthic fauna living in a small stream affected by effluent from a municipal sewage treatment plant. Water research 72: 145-153.
Guler, Y., Ford, A. T., 2010. Anti-depressants make amphipods see the light. Aquatic toxicology 99: 397-404.
Guo, J., Sinclair, C. J., Selby, K., Boxall, A. B. A., 2016. Toxicological and ecotoxicological risk-based prioritization of pharmaceuticals in the natural environment.
Environmental toxicology and chemistry. doi:10.1002/etc.3319
11
Hart Jr, C. W., Fuller, S. L. H., 1974. Pollution Ecology of Freshwater Invertebrates. New York and London: Academic Press.
Ivkovic, M., Miliša, M., Previšić, A., Popijač, A., Mihaljević, Z., 2013. Environmental control of emergence patterns: Case study of changes in hourly and dailyemergence of aquatic insects at constantand variable water temperatures. International Review of
Hydrobiology 98: 104-115.
Jiang, J., Zhou, Z., 2013. Removal of Pharmaceutical Residues by Ferrate (VI). PLoS ONE 8(2): e55729. doi:10.1371/journal.pone.0055729
Jonsson, M., Ershammar, E., Fick, J., Brodin, T., Klaminder, J., 2015. Effects of an antihistamine on carbon and nutrient recycling in streams. Science of the total environment 538: 240-245.
Klaminder, J., Brodin, T., Sundelin, A., Anderson, N. J., Fahlman, J., Jonsson, M., Fick, J., 2015. Long-Term Persistence of an Anxiolytic Drug (Oxazepam) in a Large Freshwater Lake. Environmental Science & technology 49 (17): 10406-10412.
Lürling, M., Sargant, E., Roessink, I., 2005. Life-History Consequences for Daphnia pulex Exposed to Pharmaceutical Carbamazepine. Environmental toxicology. 172-180 Marshall, E. J. R., Royer, T. V., 2012. Pharmaceutical Compounds and Ecosystem Function: An Emerging Research Challenge for Aquatic Ecosystems. Ecosystems 15:
867-880.
Nalecz-Jawecki, G., Persoone, G., 2005. Toxicity of Selected Pharmaceuticals to the Anostracan Crustacean Thamnocephalus platyurus - Comparison of Sublethal and Lethal Effect Levels with the 1h Rapidtoxkit and the 24h Thamnotoxkit Microbiotests.
Environmental Science and Pollution Research 13 (1): 22-27.
Nunes, B., Carvalho, F., Guilhermino, L., 2005. Acute toxicity of widely used pharmaceuticals in aquatic species: Gambusia holbrooki, Artemia parthenogenetica and Tetraselmis chuii. Ecotoxicology and Environmental Safety 61 (3): 413-419.
Oaks, L. J., Gilbert, M., Virani, M. Z., Watson, R. T., Meteyer, C. U., Rideout, B. A.,
Shivaprasad, H. L., Ahmed, S., Chaudhry, M. J. I., Arshad, M., Mahmood, S., Ali, A., Khan, A. A., 2004. Diclofenac residues as the cause of vulture population decline in Pakistan. Nature 427: 630-633.
Rodríguez, E. M., Márquez, G., León, E. A., Álcarez, P. M., Amat, A. M., Beltrán, F. J., 2013 Mechanism considerations for photocatalytic oxidation, ozonation and photocatalytic ozonation of some pharmaceturical compounds in water. Journal of Environmental Management 127: 114-124
Rowland, M., Noe, C. R., Smith, D. A., Tucker, G. T., Crommelin, D. J. A., Peck, C. C., Rocci Jr, M. L., Besancon, L., Shah, V. P., 2012. Impact of the Pharmaceutical Sciences on Health Care: A Reflection over the Past 50 Years. Journal of pharmaceutical science 101 (11): 4075-4099.
Rubach, M. N., Ashauer. R., Buchwalter, D. B., De Lange, H. J., Hamer, M., Preuss, T. G., Töpke, K., Maund, S. J., 2011. Framework for Traits-Based Assessment in
Ecotoxicology. Integrated Environmental Assessment and Management 7 (2): 172- 186.
Sánchez-Argüello, P., Fernández, C., Tarazona, J. V., 2009. Assessing the effects of fluoxetine on Physa acuta (Gastropoda, Pulmonata) and Chironomus riparius (Insecta, Diptera)
12
using a two-species water-sediment test. Science of the Total Environment 407: 1937- 1946.
Sumpter, John. P., 2016. Protecting aquatic organisms from chemicals: the harsh realities.
The royal society publishing 367: 3877-3894.
Water research center. 2014. Ozonation in Water Treatment. http://www.water- research.net/index.php/ozonation (Accessed 2016-05-03).
Webster, E., Mackay, D., Wania, F., 1998. Evaluating environmental persistence.
Environmental toxicology and chemistry 17 (11): 2148-2158.
Williams, M. W., Carter, L. J., Fussell, R., Raffaelli, D., Ashauer, R., Boxall A. B. A., 2011.
Uptake and depuration of pharmaceuticals in aquatic invertebrates. Environmental Pollution 165: 250-258.
Appendix 1
Table 1. Coordinates in DMS format (degrees minutes seconds) for sampling sites 5-10 in river Knivsta.
Sampling site Latitude (DMS) Longitude (DMS)
5 59 43'05.8'' 17 47'28.1''
6 59 43'4.1'' 17 47'29.2''
7 59 43'0.4'' 17 47'32.2''
8 59 42'17.5'' 17 47'31.7''
9 59 41'11.9'' 17 47'02.7''
10 59 39'38.7'' 17 45'06.8''
Appendix 2
Table 2. Number of individuals belonging to each species group and the total number of individuals found both before and during ozonation treatment in river Knivsta.
Taxa Before During
Chironomidae (Nonbiting midges) 604 995
Isopoda 138 170
Ephemeroptera (Mayfly) 22 11
Trichoptera (Caddisfly) 101 99
Zygoptera (Damselfly) 0 1
Odonata (Dragonfly) 2 0
Simuliidae (Black fly) 42 94
Plecoptera (Stonefly) 19 2
Amphipoda 33 55
Bivalvia (Mussel) 291 162
Coleoptera (Beetle) 2 2
Copepoda 22 4
Acari (Tick & mite) 9 2
Hirudinea (Leech) 92 117
Calopterygidae (Broad-winged damselfly) 5 16
Gastropoda (Snail & slug) 6 44
Tipulidae (Crane fly) 0 4
Corixidae (Water boatmen) 0 3
Lepidoptera (Moth & butterfly) 0 6
Nepidae (Water scorpions) 0 1
Total number of individuals 1388 1788
Dept. of Ecology and Environmental Science (EMG) S-901 87 Umeå, Sweden
Telephone +46 90 786 50 00 Text telephone +46 90 786 59 00 www.umu.se
