Development of fish models for assessment of chemicals, which may interfere with different parts ... more Development of fish models for assessment of chemicals, which may interfere with different parts of the life cycle, act over consecutive generations and with a potential to have impact on populations, are essential for risk assessment and environmental protection.
Archives of Environmental Contamination and Toxicology, Jul 3, 2006
Juvenile zebrafish (Danio rerio) were exposed to different dilutions (0, 0.67, 2.5, 10, and 50%) ... more Juvenile zebrafish (Danio rerio) were exposed to different dilutions (0, 0.67, 2.5, 10, and 50%) of effluent water from a Swedish pulp mill that previously has been reported to be androgenic to fish. Exposure was performed between days 10-38 days post-hatch. Fish were sampled for whole-body vitellogenin concentrations at day 38 post-hatch and for histological examination of gonads at day 60 post-hatch. In fish exposed to the highest concentration of pulp mill effluent, elevated concentrations of vitellogenin were measured. The androgenicity of the pulp mill water was confirmed by the increased number of males recorded at 60 days post-hatch. Image analysis of testes indicated stimulation of spennato genesis. Intersex fish were observed in all exposure groups. An androgenic activity equivalent to 5.6 ng/L dihydroxytestosterone was measured using the yeast androgen screen (YAS) assay. The present study demonstrates that both androgenic and estrogenic effects can be detected when exposing zebrafish during the juvenile period to complex mixtures of chemicals.
The accumulation and effects of 150 pg A1 1 ' on minnows (Phoxinusphoxinus L.) were studied in so... more The accumulation and effects of 150 pg A1 1 ' on minnows (Phoxinusphoxinus L.) were studied in soft water at pH 7 , 6 and 5. The fish were kept in a flow-through water system for up to 48 days. Addition ofaluminium to the water resulted in poor appetite, passivity and a tendency for the fish to gather as far as possible from the aluminium inlet. The mortality was high at pH 5 and also after addition of aluminium at pH 6. Severe lesions were noted in the gills and olfactory organs after exposure to aluminium. The concentration of aluminium in the gills, liver and kidney was increased at low pH. A large proportion of the aluminium was located on the surface of the gill epithelium. The recovery capacity was pronounced when the aluminium exposure was terminated and the pH was increased from 5 to 7. No further mortality occurred and gill and olfactory organ structure returned to near normality in about 36 days.
Surface water is an important resource for drinking water, irrigation water and recreation. It is... more Surface water is an important resource for drinking water, irrigation water and recreation. It is therefore necessary to protect it from pollution deteriorating its quality. Whereas several environmental directives have been implemented at the European level, e.g. Nitrate Directive (91/676/EEC) and Water Framework Directive (2000/60/EC), animal and human health is poorly protected. The Bathing Water Directive (2006/7/EC) was implemented in order to reduce human exposure to potential hazards, but rather than being a tool to improve water quality it is used merely as a means to locate public beaches and their access. Agricultural practices (e.g. manure and sludge fertilisation, animal grazing) and wastewater outlets are the main sources of faecal pollution of surface water, along with risks for further transmission of human and animal gastro-intestinal diseases. Therefore, biosecurity aspects were included in the Baltic Compass project as an important component of sustainable agriculture. This report summarises the main messages on biosecurity risks within the Baltic Sea drainage area, and how these risks can be reduced by best management practices.
The establishment of stormwater ponds for treatment of runoff water can be beneficial for amphibi... more The establishment of stormwater ponds for treatment of runoff water can be beneficial for amphibians due to creations of new habitats which often develop vegetative characteristics similar to that of natural wetlands. These ponds might help to promote stability in amphibian populations if the benefits of creation of new habitats exceed the risk for toxic impact. In the present study, Swedish stormwater ponds were searched for larval amphibians. Further, the impact of stormwater quality on embryo development was tested using Xenopus (Silurana) tropicalis. Data suggest that most of the studied stormwater ponds had water of proper quality for promoting amphibian embryo development. However there were sites in which the water caused premature hatching and/or reduced heart rate in X. tropicalis embryos. Pollutant toxicity might be one explanation for sublethal effects caused by some of the stormwater samples. In a majority of ponds, no amphibian larvae were detected and this may partly be explained by toxic compounds in the waters.
Archives of Environmental Contamination and Toxicology, Nov 24, 2006
The phthalate esters are a group of industrial chemicals considered to have endocrine-disrupting ... more The phthalate esters are a group of industrial chemicals considered to have endocrine-disrupting properties. The most common tonnage product among these, di-2-ethylhexyl phthalate (DEHP), is widely spread in the environment. The objectives with the present work were to study uptake and metabolism of orally administered DEHP and its major metabolite mono-2-ethyl hexyl phthalate (MEHP) and to evaluate the impact of early life exposure on sex differentiation in Atlantic salmon. The feeding with contaminated diet started immediately after yolk sac resorption and continued for 4 weeks. Nominal concentrations of DEHP in the diet were 400 (measured 359), 800 (measured 827), and 1500 (measured 1648) mg DEHP/kg and a control group was fed food mixed with solvent. After the exposure period, fish were fed non-contaminated diet until final sampling 4 months postexposure. There were no effects on growth or survival of the fish and no late effects on hepatosomatic index or sex ratio. However, the histological examination of gonads from fish exposed to 1500 mg DEHP/kg revealed a small but significant incidence (3%) of intersex fish (ovo-testis). Chemical residues of DEHP and MEHP were analyzed weekly during the first 3 months of the post-exposure period. Both DEHP and MEHP were rapidly eliminated to near background levels within one week post exposure. The study indicates that exposure of Atlantic salmon to relatively high concentrations of DEHP during a sensitive part of the life cycle may interfere with gonad differentiation. Environmental pollutants that can interfere with endocrine functions in animals have raised concern during the last decades. These so-called endocrine-disrupting chemicals include compounds with the ability to mimic natural hormones, block the action of natural hormones, or interfere with the synthesis and/or degradation of natural hormones (Soto et al. 1995). A group of widely used industrial chemicals considered to have endocrine-disrupting properties are the phthalate esters. These chemicals mainly serve as important additives in polyvinylchloride (PVC) resins and are used as plasticizers in various products, e.g., industrial hardware, food packaging, medical products, and toys (Staples et al. 1997; Balafas et al. 1999; Wahl et al. 1999; Earls et al. 2003). Since the phthalate esters are not chemically bound to the plastic material, they can leach into the environment (Page and Lacroix 1995). The most common tonnage product among the phthalate esters, di-2ethylhexyl phthalate (DEHP), has been found in both fresh and marine water environments (Giam et al. 1975; Tan 1995) as well as in food (Page and Lacroix 1995) and considerable concern has been focused on its potential risk to human and wild life health. DEHP, via its metabolite mono-2-ethyl hexyl phthalate (MEHP), has been shown to be a peroxisome proliferator (PP) by activating peroxisome proliferator-activated receptors in rodents (PPARs) (e.g., Moody and Reddy 1978; Lapinskas et al. 2005) and long-term exposure of rodents to DEHP has resulted in development of hepatocarcinomas (Reddy and Lalwani 1983). In fish, peroxisome proliferation has been reported after exposure to phthalates (i.e., dibutylphthalate) (Ortiz-Zarragoitia and Cajaraville 2000) and PPARs with similar sequence and distribution as in mammals have recently been found in a few fish species, including Atlantic salmon (Salmo salar) (Ruyter et al. 1997; Andersen et al. 2000). Effects of DEHP on reproduction and development have been shown in several vertebrates but the mechanism of action remains unclear. The estrogenic activity of DEHP has been tested in different in vitro assays showing weak or no affinity to the estrogen receptor (ER) at high concentrations (
Clozapine (an atypical antipsychotic drug) is indicated in treatment-refractory schizophrenia. An... more Clozapine (an atypical antipsychotic drug) is indicated in treatment-refractory schizophrenia. Antipsychotics have been reported to have cardiotoxic effects in humans.The zebra fish (Danio rerio) is a tropical cyprinid used in toxicological research. In the present study, the toxicity of clozapine was evaluated in zebra fish embryos with the aid of the fish embryo toxicity test (FET). The endpoints monitored included circulation, heart rate, pericardial oedema, tail extension, spinal deformation, hatching time, coagulation and death. Clozapine was tested at five different concentrations ranging from 1 µg/L to 10 mg/L. The toxic effects observed were a marked bradycardia at 10 mg/L and various abnormalities in the zebra fish embryos exposed to clozapine at 1 µg/L to 10 mg/L. Based on the results obtained in this study, clozapine is toxic to zebra fish embryos and its level in the aquatic environment should be closely monitored in order to forestall adverse effects on the reproductive capabilities and survival of aquatic organisms.
... The fish also had diminished body weight gain, poor food conversion, low hema-tocrit, decreas... more ... The fish also had diminished body weight gain, poor food conversion, low hema-tocrit, decreased hepatosomatic index, as well as reduced ... 6. Parihar, MS, Dubey, AK, Javeri, T, and Prakash, P., Changes in lipid peroxida-tion, superoxide dismutase activity, ascorbic acid and ...
Toxicity tests of musk ketone (MK) and tetrabromobisphenol-A (TBBPA) on embryos were conducted in... more Toxicity tests of musk ketone (MK) and tetrabromobisphenol-A (TBBPA) on embryos were conducted in two amphibian species, Xenopus (Silurana) tropicalis and the Swedish native species Rana arvalis. TBBPA was also tested on fish embryos of Danio rerio. All species were tested in similar experimental setup. Musk ketone caused decreased heart rates at concentrations from 10 and 100 g/L in R. arvalis and X. tropicalis, respectively. TBBPA caused effects at 1000 g/L in all three species. The responses were comparable between all three species which supports the relevance for using data from non-native species in national risk assessment.
Environmental Toxicology and Chemistry, Nov 1, 1993
In 1974, abnormally high mortality was recorded among yolk-sac fry of Baltic salmon (Salmo salar)... more In 1974, abnormally high mortality was recorded among yolk-sac fry of Baltic salmon (Salmo salar) originating from feral females manually stripped and fertilized with milt from feral males. The cause of this mortality, designated M74, is unknown. Our hypothesis is that xenobiotic compounds responsible for reproduction failure in higher vertebrates in the Baltic Sea also interfere with reproduction in Baltic salmon. The significance of M74 should not be underestimated, because the syndrome has caused up to 75% yearly mortality of developing Baltic salmon yolk-sac larvae in a fish hatchery dedicated to production of smolt during the last two decades. We cannot exclude the possibility that only a relatively low number of naturally spawned eggs develop normally because of M74. No individual pollutant has been shown to be responsible for the development of M74 syndrome. However, a higher total body burden of organochlorine substances may be responsible for the M74 syndrome. The presence of induced hepatic cytochrome P450 enzymes in both yolk-sac fry suffering from M74 and adult feral females producing offspring affected by M74 supports this hypothesis. In addition, the P450 enzyme activity in offspring from feral fish is higher than the activity in yolk-sac fry from hatchery-raised fish, suggesting that feral Baltic salmon are influenced by organic xenobiotics.
Comparative Biochemistry and Physiology C-toxicology & Pharmacology, Sep 1, 2006
Managed by the Organisation for Economic Co-operation and Development (OECD), a comprehensive wor... more Managed by the Organisation for Economic Co-operation and Development (OECD), a comprehensive work is carried out in numerous laboratories to develop test guidelines for the detection of endocrine disrupting chemicals in humans, and various animal species. Development of tests to detect chemicals with endocrine disrupting properties in fish is a part of that work. A Fish Sexual Development Test (FSDT) (an extension of the existing OECD TG 210, fish early life stage toxicity test), proposed as an international test guideline for the detection of endocrine disrupting chemicals, was evaluated by water exposure of juvenile zebrafish to the three natural estrogens: estrone, 17beta-estradiol, and estriol and the synthetic androgen trenbolone (trenbolone acetate). As endpoints, vitellogenin induction and histological changes including changes in sex ratios were investigated. The sex ratio was significantly altered towards females from 49 ng/l estrone, 54 ng/l 17beta-estradiol and 22 microg/l estriol, respectively. An all male population was observed from exposure to 9.7 ng/l trenbolone and above. Significant vitellogenin induction in whole body homogenate was measured after exposure to 14 ng/l estrone, 54 ng/l 17beta-estradiol and 0.6 mug/l estriol, respectively. Significant vitellogenin reduction was measured after exposure to 193 ng/l trenbolone or higher. The present results provide strong evidence that the FSDT is a sensitive test toward estrogenic and especially androgenic exposure and the validation of the FSDT as an OECD test guideline should continue.
Female three-spined sticklebacks were fed with freeze-dried chironomids contaminated with low or ... more Female three-spined sticklebacks were fed with freeze-dried chironomids contaminated with low or high doses of polybrominated diphenyl ethers (PBDEs) (Bromkal 70-5DE), polychlorinated naphthalenes (PCNs) (Halowax 1014) and polychlorinated biphenyls (PCBs) (Clophen A50). After 3.5 months of exposure, reproduction studies were started. No significant difference in number of eggs laid was found between the control and exposed groups. Spawning success (considered successful if it occurred within 24 h) in the control group was 80%, whereas it was 20% and 25% in the groups that received high doses of Bromkal 70-5DE or Clophen A50, respectively. After spawning, the fish were dissected for chemical, biochemical and morphological analyses. Levels of PBDE in the low-and high-dose Bromkal 70-5DE groups were 861 + 271 and 1630 + 275 mg/kg fat, respectively, whereas the corresponding concentrations of PCN in the Halowax 1014 groups were 845 + 43 and 1929 + 72 mg/kg fat, respectively. Concentrations of PCB in fish from the Clophen A50 groups were 1972 + 158 and 3594 + 521 mg/kg fat, respectively. Uptake efficiency was approx. 40% for PCB and 20% for PBDE and PCN. Hepatic cytochrome P450-dependent ethoxyresorufin-O-deethylase (EROD) activity was induced by Clophen A50 and Halowax 1014, whereas Bromkal 70-5DE did not significantly induce the enzyme system. EROD activity was 0.8 pmol/mg prot./min in the control group, whereas it was between 2.7 and 16.3 pmol/mg prot./min in the exposed groups. Furthermore, the morphological examination of the liver disclosed pronounced lipid accumulation in all exposed groups.
Page 1. Jenny Lundstrom, Bethanie Carney, Patric Amcoff, Annette Pettersson, Hans Borjeson, Lars ... more Page 1. Jenny Lundstrom, Bethanie Carney, Patric Amcoff, Annette Pettersson, Hans Borjeson, Lars Forlin and Leif Norrgren ... pollution of the Baltic Sea has been considered as a key causative factor for reproduction disturbances in mammalian (1, 2), bird (3), and fish (4) species ...
Peri-urban water bodies are at risk from excessive pollution as they are direct sinks for urban e... more Peri-urban water bodies are at risk from excessive pollution as they are direct sinks for urban effluents. The occurrence of oestrogenic and androgenic endocrine disrupting chemicals (EDCs) in effluents and water bodies around the city of Bulawayo, Zimbabwe, and their effects on wild fish was investigated in 2013. Effluent and water were sampled from sewage treatment plants, an urban stream, and effluent-polluted dams, and were compared with a 'pristine' dam upstream of Bulawayo. Organic pollutants were extracted by solid-phase extraction and tested for EDCs using a yeast oestrogen/androgen screen. Oestrogenic and androgenic potencies were expressed as 17β-oestradiol equivalent (EEq) or dihydrotestosterone equivalent (TEq). Tilapia and catfish from the dams were analysed for gonado-somatic indices and testis histopathology. Effluents from STPs, which directly flow into Umguza Dam, had EEq of 33 ng l −1 and 55 ng l −1 , respectively. Umguza Dam, Khami Dam and Matsheumhlope Stream had EEqs of 237 ng l-1 , 9 ng l −1 and 2 ng l −1 , respectively. Androgenic activity was detected in only one STP (TEq = 93 ng l −1). Tilapia sampled from effluent-polluted dams had high incidences of testis-ova, but catfish had no signs of reproductive dysfunction. These findings underscore the need for greater attention to EDCs in developing countries where there is scant literature regarding their occurrence and impacts.
Aquatic Ecosystem Health & Management, Sep 1, 2001
Morphological studies of gonads from roach (Rutilus rutilus) in a small Swedish lake were perform... more Morphological studies of gonads from roach (Rutilus rutilus) in a small Swedish lake were performed. The lake is a potential recipient for drainage water from a refuse dumping area. All fishes were macroscopically identified as males, but histological examination of the gonads showed a high incidence of intersex. This high incidence might be caused by endocrine disrupting chemicals. The relationships between exposure to endocrine disrupters and intersex observations in wild fish is unknown and the effect of endocrine disrupters on zebrafish (Danio rerio) were evaluated in order to determine whether they might serve as a model laboratory species for effects of endocrine disrupting chemicals in wild fish. Zebrafish were exposed from pre-blastula stage until sexual maturation to either standardised water (controls), 17a-methyltestosterone (1, 10 μg l-1) or 17b-estradiol (1, 10 μg l-1). Exposure to both concentrations of 17bestradiol caused a significant feminisation, while a significant increase in the proportions of males was found after exposure to 1 mg 17a-methyltestosterone l-1. Furthermore, exposure to 1 and 10 mg 17a-methyltestosterone l-1 caused development of intersex individuals. Additionally, sexually mature male zebrafish were exposed to 1 μg 17b-estradiol l-1. After 10 weeks of exposure the fish were transferred to non-contaminated water for two weeks. The control group was kept in standardised water for 12 weeks. At termination of the experiment, measurement of whole-body vitellogenin concentrations were performed. Exposure to 17b-estradiol caused a significant increase in whole body vitellogenin concentrations. After 2 weeks in water without addition of 17b-estradiol, the vitellogenin concentration was reduced by approximately 50%. Roach and zebrafish may share properties which makes these species suitable for further comparisons to evaluate effects of endocrine disrupting chemicals as well as domestic and industrial effluents containing hormone-like substances.
This study investigated the suitability of using tadpoles of the West-African clawed frog (Xenopu... more This study investigated the suitability of using tadpoles of the West-African clawed frog (Xenopus tropicalis) for studying adverse effects on the thyroid hormone system after chemical exposure. Tadpoles were exposed to the thyroxine synthesis inhibitor 6-propylthiouracil (PTU) at concentrations between 2-75 mg/L during 14 days. After 5 and 14 days of exposure the developmental stage, hind limb length, body length and weight were measured. Moreover, histological measurements of the thyroid glands were performed after 14 days of exposure. These measurements included maximum thyroid cross-section area, follicular area and epithelial cell height. Tadpoles in the 75 mg/L treatment were less developed and had shorter hind limb length than the control group after 14 days of exposure. No effects were recorded on these parameters at lower PTU concentrations. The histological measurements revealed clear dose-response relationships in both follicular cross-section area and epithelial cell height, with lowest observed effect concentrations (LOECs) recorded at 2 and 5 mg/L, respectively. This study shows that X. tropicalis is a suitable species for detection of thyroid disrupting chemicals. Further, histopathological measurements of thyroid glands are more sensitive parameters compared with apical endpoints when studying adverse effects on thyroid hormone system caused by PTU exposure in X. tropicalis.
The high concentrations of polybrominated diphenylethers (PBDEs) in the environment have raised t... more The high concentrations of polybrominated diphenylethers (PBDEs) in the environment have raised the need for generating more information about the impact of these substances on animals. To study the distribution of 14 C-labelled 2,2 ,4,4 ,5-pentabromodiphenyl ether (14 C-BDE-99) in Xenopus tropicalis (West African clawed frog) 14 C-BDE-99 was administered by dietary exposure to tadpoles at stage 54 or to juvenile frogs at stage 66. Whole-body autoradiography and liquid scintillation counting were used to examine the distribution of the substance at different survival times. Further, X. tropicalis tadpoles were dietarily exposed to the PBDE congeners BDE-47 and BDE-99 to study the effects on metamorphosis process. Measurements like body weight, body length, hind limb length and developmental stage as well as histological measurements on thyroid glands were performed after 14 days of exposure. Autoradiograms revealed high concentrations and long term retention of 14 C-BDE-99 in adipose tissue and melanin in frogs exposed both as tadpoles and juveniles. Further, a difference in uptake was recorded between the exposures at stages 54 and 66, implying that the juvenile frogs have higher uptake and more prolonged retention of the chemical than the tadpoles. Hind limb length was reduced in tadpoles dietarily exposed to 1 mg/g feed of both BDE congeners. This was associated with reduced body weight and body length for BDE-47, suggesting general toxicity. Tadpoles exposed to BDE-99 also showed lower developmental stage but no effects on body weight or body length, suggesting possible thyroid hormone disruption. Higher concentrations of both congeners caused increased mortality. Thus, it can be concluded that in the present study, BDE-99 was retained for a longer period in the juvenile frogs than in metamorphosing tadpoles and that BDE-99 had an impact on X. tropicalis metamorphosis that might be of thyroid disrupting origin.
Archives of Environmental Contamination and Toxicology, Apr 1, 2000
Zambia is a country with an extensive mining industry with the majority of mines located in the C... more Zambia is a country with an extensive mining industry with the majority of mines located in the Copperbelt province. Through this region of the country, the Kafue River drains and receives effluent water from mining activities as well as from other industrial point sources. In addition, production of agricultural products and pest control requires use of different pesticides in the area. Information on industrial and agricultural pollution has not been clearly identified in Zambia, and little attention has been paid to pollution control and possible impact of metals, pesticides, and other persistent compounds in the environment. The objective of this study was to introduce and to evaluate a few methodologies based on in situ bioassays for environmental assessment to promote sustainable and environmentally sound water resource management of the Kafue River. The results show that caged threespot tilapia exposed downstream of industrial points sources rapidly bioaccumulate several trace elements, i.e., Cd, Co, Cu, Cr, Ni. These elements also occurred in much higher concentrations in water samples downstream of the industrial area compared with a locality upstream. Furthermore, the use of a semipermeable membrane device (SPMD) for passive absorption of lipophilic pollutants in the water showed relatively high concentration of several pesticides, i.e., DDT with major metabolites, PCB, and dieldrin. The present study shows that only 2 weeks of in situ studies in waters contaminated by pollutants affects in situ exposed fish and that the correlation between water and tissue concentrations was relatively good. Both trace elements and persistent organic pollutants occurred in such high concentrations that they must be considered from ecotoxicological aspects and may affect aquatic animal health.
Development of fish models for assessment of chemicals, which may interfere with different parts ... more Development of fish models for assessment of chemicals, which may interfere with different parts of the life cycle, act over consecutive generations and with a potential to have impact on populations, are essential for risk assessment and environmental protection.
Archives of Environmental Contamination and Toxicology, Jul 3, 2006
Juvenile zebrafish (Danio rerio) were exposed to different dilutions (0, 0.67, 2.5, 10, and 50%) ... more Juvenile zebrafish (Danio rerio) were exposed to different dilutions (0, 0.67, 2.5, 10, and 50%) of effluent water from a Swedish pulp mill that previously has been reported to be androgenic to fish. Exposure was performed between days 10-38 days post-hatch. Fish were sampled for whole-body vitellogenin concentrations at day 38 post-hatch and for histological examination of gonads at day 60 post-hatch. In fish exposed to the highest concentration of pulp mill effluent, elevated concentrations of vitellogenin were measured. The androgenicity of the pulp mill water was confirmed by the increased number of males recorded at 60 days post-hatch. Image analysis of testes indicated stimulation of spennato genesis. Intersex fish were observed in all exposure groups. An androgenic activity equivalent to 5.6 ng/L dihydroxytestosterone was measured using the yeast androgen screen (YAS) assay. The present study demonstrates that both androgenic and estrogenic effects can be detected when exposing zebrafish during the juvenile period to complex mixtures of chemicals.
The accumulation and effects of 150 pg A1 1 ' on minnows (Phoxinusphoxinus L.) were studied in so... more The accumulation and effects of 150 pg A1 1 ' on minnows (Phoxinusphoxinus L.) were studied in soft water at pH 7 , 6 and 5. The fish were kept in a flow-through water system for up to 48 days. Addition ofaluminium to the water resulted in poor appetite, passivity and a tendency for the fish to gather as far as possible from the aluminium inlet. The mortality was high at pH 5 and also after addition of aluminium at pH 6. Severe lesions were noted in the gills and olfactory organs after exposure to aluminium. The concentration of aluminium in the gills, liver and kidney was increased at low pH. A large proportion of the aluminium was located on the surface of the gill epithelium. The recovery capacity was pronounced when the aluminium exposure was terminated and the pH was increased from 5 to 7. No further mortality occurred and gill and olfactory organ structure returned to near normality in about 36 days.
Surface water is an important resource for drinking water, irrigation water and recreation. It is... more Surface water is an important resource for drinking water, irrigation water and recreation. It is therefore necessary to protect it from pollution deteriorating its quality. Whereas several environmental directives have been implemented at the European level, e.g. Nitrate Directive (91/676/EEC) and Water Framework Directive (2000/60/EC), animal and human health is poorly protected. The Bathing Water Directive (2006/7/EC) was implemented in order to reduce human exposure to potential hazards, but rather than being a tool to improve water quality it is used merely as a means to locate public beaches and their access. Agricultural practices (e.g. manure and sludge fertilisation, animal grazing) and wastewater outlets are the main sources of faecal pollution of surface water, along with risks for further transmission of human and animal gastro-intestinal diseases. Therefore, biosecurity aspects were included in the Baltic Compass project as an important component of sustainable agriculture. This report summarises the main messages on biosecurity risks within the Baltic Sea drainage area, and how these risks can be reduced by best management practices.
The establishment of stormwater ponds for treatment of runoff water can be beneficial for amphibi... more The establishment of stormwater ponds for treatment of runoff water can be beneficial for amphibians due to creations of new habitats which often develop vegetative characteristics similar to that of natural wetlands. These ponds might help to promote stability in amphibian populations if the benefits of creation of new habitats exceed the risk for toxic impact. In the present study, Swedish stormwater ponds were searched for larval amphibians. Further, the impact of stormwater quality on embryo development was tested using Xenopus (Silurana) tropicalis. Data suggest that most of the studied stormwater ponds had water of proper quality for promoting amphibian embryo development. However there were sites in which the water caused premature hatching and/or reduced heart rate in X. tropicalis embryos. Pollutant toxicity might be one explanation for sublethal effects caused by some of the stormwater samples. In a majority of ponds, no amphibian larvae were detected and this may partly be explained by toxic compounds in the waters.
Archives of Environmental Contamination and Toxicology, Nov 24, 2006
The phthalate esters are a group of industrial chemicals considered to have endocrine-disrupting ... more The phthalate esters are a group of industrial chemicals considered to have endocrine-disrupting properties. The most common tonnage product among these, di-2-ethylhexyl phthalate (DEHP), is widely spread in the environment. The objectives with the present work were to study uptake and metabolism of orally administered DEHP and its major metabolite mono-2-ethyl hexyl phthalate (MEHP) and to evaluate the impact of early life exposure on sex differentiation in Atlantic salmon. The feeding with contaminated diet started immediately after yolk sac resorption and continued for 4 weeks. Nominal concentrations of DEHP in the diet were 400 (measured 359), 800 (measured 827), and 1500 (measured 1648) mg DEHP/kg and a control group was fed food mixed with solvent. After the exposure period, fish were fed non-contaminated diet until final sampling 4 months postexposure. There were no effects on growth or survival of the fish and no late effects on hepatosomatic index or sex ratio. However, the histological examination of gonads from fish exposed to 1500 mg DEHP/kg revealed a small but significant incidence (3%) of intersex fish (ovo-testis). Chemical residues of DEHP and MEHP were analyzed weekly during the first 3 months of the post-exposure period. Both DEHP and MEHP were rapidly eliminated to near background levels within one week post exposure. The study indicates that exposure of Atlantic salmon to relatively high concentrations of DEHP during a sensitive part of the life cycle may interfere with gonad differentiation. Environmental pollutants that can interfere with endocrine functions in animals have raised concern during the last decades. These so-called endocrine-disrupting chemicals include compounds with the ability to mimic natural hormones, block the action of natural hormones, or interfere with the synthesis and/or degradation of natural hormones (Soto et al. 1995). A group of widely used industrial chemicals considered to have endocrine-disrupting properties are the phthalate esters. These chemicals mainly serve as important additives in polyvinylchloride (PVC) resins and are used as plasticizers in various products, e.g., industrial hardware, food packaging, medical products, and toys (Staples et al. 1997; Balafas et al. 1999; Wahl et al. 1999; Earls et al. 2003). Since the phthalate esters are not chemically bound to the plastic material, they can leach into the environment (Page and Lacroix 1995). The most common tonnage product among the phthalate esters, di-2ethylhexyl phthalate (DEHP), has been found in both fresh and marine water environments (Giam et al. 1975; Tan 1995) as well as in food (Page and Lacroix 1995) and considerable concern has been focused on its potential risk to human and wild life health. DEHP, via its metabolite mono-2-ethyl hexyl phthalate (MEHP), has been shown to be a peroxisome proliferator (PP) by activating peroxisome proliferator-activated receptors in rodents (PPARs) (e.g., Moody and Reddy 1978; Lapinskas et al. 2005) and long-term exposure of rodents to DEHP has resulted in development of hepatocarcinomas (Reddy and Lalwani 1983). In fish, peroxisome proliferation has been reported after exposure to phthalates (i.e., dibutylphthalate) (Ortiz-Zarragoitia and Cajaraville 2000) and PPARs with similar sequence and distribution as in mammals have recently been found in a few fish species, including Atlantic salmon (Salmo salar) (Ruyter et al. 1997; Andersen et al. 2000). Effects of DEHP on reproduction and development have been shown in several vertebrates but the mechanism of action remains unclear. The estrogenic activity of DEHP has been tested in different in vitro assays showing weak or no affinity to the estrogen receptor (ER) at high concentrations (
Clozapine (an atypical antipsychotic drug) is indicated in treatment-refractory schizophrenia. An... more Clozapine (an atypical antipsychotic drug) is indicated in treatment-refractory schizophrenia. Antipsychotics have been reported to have cardiotoxic effects in humans.The zebra fish (Danio rerio) is a tropical cyprinid used in toxicological research. In the present study, the toxicity of clozapine was evaluated in zebra fish embryos with the aid of the fish embryo toxicity test (FET). The endpoints monitored included circulation, heart rate, pericardial oedema, tail extension, spinal deformation, hatching time, coagulation and death. Clozapine was tested at five different concentrations ranging from 1 µg/L to 10 mg/L. The toxic effects observed were a marked bradycardia at 10 mg/L and various abnormalities in the zebra fish embryos exposed to clozapine at 1 µg/L to 10 mg/L. Based on the results obtained in this study, clozapine is toxic to zebra fish embryos and its level in the aquatic environment should be closely monitored in order to forestall adverse effects on the reproductive capabilities and survival of aquatic organisms.
... The fish also had diminished body weight gain, poor food conversion, low hema-tocrit, decreas... more ... The fish also had diminished body weight gain, poor food conversion, low hema-tocrit, decreased hepatosomatic index, as well as reduced ... 6. Parihar, MS, Dubey, AK, Javeri, T, and Prakash, P., Changes in lipid peroxida-tion, superoxide dismutase activity, ascorbic acid and ...
Toxicity tests of musk ketone (MK) and tetrabromobisphenol-A (TBBPA) on embryos were conducted in... more Toxicity tests of musk ketone (MK) and tetrabromobisphenol-A (TBBPA) on embryos were conducted in two amphibian species, Xenopus (Silurana) tropicalis and the Swedish native species Rana arvalis. TBBPA was also tested on fish embryos of Danio rerio. All species were tested in similar experimental setup. Musk ketone caused decreased heart rates at concentrations from 10 and 100 g/L in R. arvalis and X. tropicalis, respectively. TBBPA caused effects at 1000 g/L in all three species. The responses were comparable between all three species which supports the relevance for using data from non-native species in national risk assessment.
Environmental Toxicology and Chemistry, Nov 1, 1993
In 1974, abnormally high mortality was recorded among yolk-sac fry of Baltic salmon (Salmo salar)... more In 1974, abnormally high mortality was recorded among yolk-sac fry of Baltic salmon (Salmo salar) originating from feral females manually stripped and fertilized with milt from feral males. The cause of this mortality, designated M74, is unknown. Our hypothesis is that xenobiotic compounds responsible for reproduction failure in higher vertebrates in the Baltic Sea also interfere with reproduction in Baltic salmon. The significance of M74 should not be underestimated, because the syndrome has caused up to 75% yearly mortality of developing Baltic salmon yolk-sac larvae in a fish hatchery dedicated to production of smolt during the last two decades. We cannot exclude the possibility that only a relatively low number of naturally spawned eggs develop normally because of M74. No individual pollutant has been shown to be responsible for the development of M74 syndrome. However, a higher total body burden of organochlorine substances may be responsible for the M74 syndrome. The presence of induced hepatic cytochrome P450 enzymes in both yolk-sac fry suffering from M74 and adult feral females producing offspring affected by M74 supports this hypothesis. In addition, the P450 enzyme activity in offspring from feral fish is higher than the activity in yolk-sac fry from hatchery-raised fish, suggesting that feral Baltic salmon are influenced by organic xenobiotics.
Comparative Biochemistry and Physiology C-toxicology & Pharmacology, Sep 1, 2006
Managed by the Organisation for Economic Co-operation and Development (OECD), a comprehensive wor... more Managed by the Organisation for Economic Co-operation and Development (OECD), a comprehensive work is carried out in numerous laboratories to develop test guidelines for the detection of endocrine disrupting chemicals in humans, and various animal species. Development of tests to detect chemicals with endocrine disrupting properties in fish is a part of that work. A Fish Sexual Development Test (FSDT) (an extension of the existing OECD TG 210, fish early life stage toxicity test), proposed as an international test guideline for the detection of endocrine disrupting chemicals, was evaluated by water exposure of juvenile zebrafish to the three natural estrogens: estrone, 17beta-estradiol, and estriol and the synthetic androgen trenbolone (trenbolone acetate). As endpoints, vitellogenin induction and histological changes including changes in sex ratios were investigated. The sex ratio was significantly altered towards females from 49 ng/l estrone, 54 ng/l 17beta-estradiol and 22 microg/l estriol, respectively. An all male population was observed from exposure to 9.7 ng/l trenbolone and above. Significant vitellogenin induction in whole body homogenate was measured after exposure to 14 ng/l estrone, 54 ng/l 17beta-estradiol and 0.6 mug/l estriol, respectively. Significant vitellogenin reduction was measured after exposure to 193 ng/l trenbolone or higher. The present results provide strong evidence that the FSDT is a sensitive test toward estrogenic and especially androgenic exposure and the validation of the FSDT as an OECD test guideline should continue.
Female three-spined sticklebacks were fed with freeze-dried chironomids contaminated with low or ... more Female three-spined sticklebacks were fed with freeze-dried chironomids contaminated with low or high doses of polybrominated diphenyl ethers (PBDEs) (Bromkal 70-5DE), polychlorinated naphthalenes (PCNs) (Halowax 1014) and polychlorinated biphenyls (PCBs) (Clophen A50). After 3.5 months of exposure, reproduction studies were started. No significant difference in number of eggs laid was found between the control and exposed groups. Spawning success (considered successful if it occurred within 24 h) in the control group was 80%, whereas it was 20% and 25% in the groups that received high doses of Bromkal 70-5DE or Clophen A50, respectively. After spawning, the fish were dissected for chemical, biochemical and morphological analyses. Levels of PBDE in the low-and high-dose Bromkal 70-5DE groups were 861 + 271 and 1630 + 275 mg/kg fat, respectively, whereas the corresponding concentrations of PCN in the Halowax 1014 groups were 845 + 43 and 1929 + 72 mg/kg fat, respectively. Concentrations of PCB in fish from the Clophen A50 groups were 1972 + 158 and 3594 + 521 mg/kg fat, respectively. Uptake efficiency was approx. 40% for PCB and 20% for PBDE and PCN. Hepatic cytochrome P450-dependent ethoxyresorufin-O-deethylase (EROD) activity was induced by Clophen A50 and Halowax 1014, whereas Bromkal 70-5DE did not significantly induce the enzyme system. EROD activity was 0.8 pmol/mg prot./min in the control group, whereas it was between 2.7 and 16.3 pmol/mg prot./min in the exposed groups. Furthermore, the morphological examination of the liver disclosed pronounced lipid accumulation in all exposed groups.
Page 1. Jenny Lundstrom, Bethanie Carney, Patric Amcoff, Annette Pettersson, Hans Borjeson, Lars ... more Page 1. Jenny Lundstrom, Bethanie Carney, Patric Amcoff, Annette Pettersson, Hans Borjeson, Lars Forlin and Leif Norrgren ... pollution of the Baltic Sea has been considered as a key causative factor for reproduction disturbances in mammalian (1, 2), bird (3), and fish (4) species ...
Peri-urban water bodies are at risk from excessive pollution as they are direct sinks for urban e... more Peri-urban water bodies are at risk from excessive pollution as they are direct sinks for urban effluents. The occurrence of oestrogenic and androgenic endocrine disrupting chemicals (EDCs) in effluents and water bodies around the city of Bulawayo, Zimbabwe, and their effects on wild fish was investigated in 2013. Effluent and water were sampled from sewage treatment plants, an urban stream, and effluent-polluted dams, and were compared with a 'pristine' dam upstream of Bulawayo. Organic pollutants were extracted by solid-phase extraction and tested for EDCs using a yeast oestrogen/androgen screen. Oestrogenic and androgenic potencies were expressed as 17β-oestradiol equivalent (EEq) or dihydrotestosterone equivalent (TEq). Tilapia and catfish from the dams were analysed for gonado-somatic indices and testis histopathology. Effluents from STPs, which directly flow into Umguza Dam, had EEq of 33 ng l −1 and 55 ng l −1 , respectively. Umguza Dam, Khami Dam and Matsheumhlope Stream had EEqs of 237 ng l-1 , 9 ng l −1 and 2 ng l −1 , respectively. Androgenic activity was detected in only one STP (TEq = 93 ng l −1). Tilapia sampled from effluent-polluted dams had high incidences of testis-ova, but catfish had no signs of reproductive dysfunction. These findings underscore the need for greater attention to EDCs in developing countries where there is scant literature regarding their occurrence and impacts.
Aquatic Ecosystem Health & Management, Sep 1, 2001
Morphological studies of gonads from roach (Rutilus rutilus) in a small Swedish lake were perform... more Morphological studies of gonads from roach (Rutilus rutilus) in a small Swedish lake were performed. The lake is a potential recipient for drainage water from a refuse dumping area. All fishes were macroscopically identified as males, but histological examination of the gonads showed a high incidence of intersex. This high incidence might be caused by endocrine disrupting chemicals. The relationships between exposure to endocrine disrupters and intersex observations in wild fish is unknown and the effect of endocrine disrupters on zebrafish (Danio rerio) were evaluated in order to determine whether they might serve as a model laboratory species for effects of endocrine disrupting chemicals in wild fish. Zebrafish were exposed from pre-blastula stage until sexual maturation to either standardised water (controls), 17a-methyltestosterone (1, 10 μg l-1) or 17b-estradiol (1, 10 μg l-1). Exposure to both concentrations of 17bestradiol caused a significant feminisation, while a significant increase in the proportions of males was found after exposure to 1 mg 17a-methyltestosterone l-1. Furthermore, exposure to 1 and 10 mg 17a-methyltestosterone l-1 caused development of intersex individuals. Additionally, sexually mature male zebrafish were exposed to 1 μg 17b-estradiol l-1. After 10 weeks of exposure the fish were transferred to non-contaminated water for two weeks. The control group was kept in standardised water for 12 weeks. At termination of the experiment, measurement of whole-body vitellogenin concentrations were performed. Exposure to 17b-estradiol caused a significant increase in whole body vitellogenin concentrations. After 2 weeks in water without addition of 17b-estradiol, the vitellogenin concentration was reduced by approximately 50%. Roach and zebrafish may share properties which makes these species suitable for further comparisons to evaluate effects of endocrine disrupting chemicals as well as domestic and industrial effluents containing hormone-like substances.
This study investigated the suitability of using tadpoles of the West-African clawed frog (Xenopu... more This study investigated the suitability of using tadpoles of the West-African clawed frog (Xenopus tropicalis) for studying adverse effects on the thyroid hormone system after chemical exposure. Tadpoles were exposed to the thyroxine synthesis inhibitor 6-propylthiouracil (PTU) at concentrations between 2-75 mg/L during 14 days. After 5 and 14 days of exposure the developmental stage, hind limb length, body length and weight were measured. Moreover, histological measurements of the thyroid glands were performed after 14 days of exposure. These measurements included maximum thyroid cross-section area, follicular area and epithelial cell height. Tadpoles in the 75 mg/L treatment were less developed and had shorter hind limb length than the control group after 14 days of exposure. No effects were recorded on these parameters at lower PTU concentrations. The histological measurements revealed clear dose-response relationships in both follicular cross-section area and epithelial cell height, with lowest observed effect concentrations (LOECs) recorded at 2 and 5 mg/L, respectively. This study shows that X. tropicalis is a suitable species for detection of thyroid disrupting chemicals. Further, histopathological measurements of thyroid glands are more sensitive parameters compared with apical endpoints when studying adverse effects on thyroid hormone system caused by PTU exposure in X. tropicalis.
The high concentrations of polybrominated diphenylethers (PBDEs) in the environment have raised t... more The high concentrations of polybrominated diphenylethers (PBDEs) in the environment have raised the need for generating more information about the impact of these substances on animals. To study the distribution of 14 C-labelled 2,2 ,4,4 ,5-pentabromodiphenyl ether (14 C-BDE-99) in Xenopus tropicalis (West African clawed frog) 14 C-BDE-99 was administered by dietary exposure to tadpoles at stage 54 or to juvenile frogs at stage 66. Whole-body autoradiography and liquid scintillation counting were used to examine the distribution of the substance at different survival times. Further, X. tropicalis tadpoles were dietarily exposed to the PBDE congeners BDE-47 and BDE-99 to study the effects on metamorphosis process. Measurements like body weight, body length, hind limb length and developmental stage as well as histological measurements on thyroid glands were performed after 14 days of exposure. Autoradiograms revealed high concentrations and long term retention of 14 C-BDE-99 in adipose tissue and melanin in frogs exposed both as tadpoles and juveniles. Further, a difference in uptake was recorded between the exposures at stages 54 and 66, implying that the juvenile frogs have higher uptake and more prolonged retention of the chemical than the tadpoles. Hind limb length was reduced in tadpoles dietarily exposed to 1 mg/g feed of both BDE congeners. This was associated with reduced body weight and body length for BDE-47, suggesting general toxicity. Tadpoles exposed to BDE-99 also showed lower developmental stage but no effects on body weight or body length, suggesting possible thyroid hormone disruption. Higher concentrations of both congeners caused increased mortality. Thus, it can be concluded that in the present study, BDE-99 was retained for a longer period in the juvenile frogs than in metamorphosing tadpoles and that BDE-99 had an impact on X. tropicalis metamorphosis that might be of thyroid disrupting origin.
Archives of Environmental Contamination and Toxicology, Apr 1, 2000
Zambia is a country with an extensive mining industry with the majority of mines located in the C... more Zambia is a country with an extensive mining industry with the majority of mines located in the Copperbelt province. Through this region of the country, the Kafue River drains and receives effluent water from mining activities as well as from other industrial point sources. In addition, production of agricultural products and pest control requires use of different pesticides in the area. Information on industrial and agricultural pollution has not been clearly identified in Zambia, and little attention has been paid to pollution control and possible impact of metals, pesticides, and other persistent compounds in the environment. The objective of this study was to introduce and to evaluate a few methodologies based on in situ bioassays for environmental assessment to promote sustainable and environmentally sound water resource management of the Kafue River. The results show that caged threespot tilapia exposed downstream of industrial points sources rapidly bioaccumulate several trace elements, i.e., Cd, Co, Cu, Cr, Ni. These elements also occurred in much higher concentrations in water samples downstream of the industrial area compared with a locality upstream. Furthermore, the use of a semipermeable membrane device (SPMD) for passive absorption of lipophilic pollutants in the water showed relatively high concentration of several pesticides, i.e., DDT with major metabolites, PCB, and dieldrin. The present study shows that only 2 weeks of in situ studies in waters contaminated by pollutants affects in situ exposed fish and that the correlation between water and tissue concentrations was relatively good. Both trace elements and persistent organic pollutants occurred in such high concentrations that they must be considered from ecotoxicological aspects and may affect aquatic animal health.
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