Natural mercury exposure of European insectivorous bats may exceed a recognized toxicity threshold.

Heavy metals are an important group of toxic substances harmful for many organisms. Of these, mercury is one of the most monitored in the environment. Several matrices are used for the monitoring of environmental load, including a range of organisms; bats, however, have only been examined rarely. Insectivorous bats are apex predators threatened by several human interventions in their natural environment, including heavy metal pollution. The aim of this study was to analyze the content of total mercury in the fur, flight membrane, and pectoral muscle of greater mouse-eared bats (Myotis myotis). Total mercury concentrations were also measured in carabid beetles from the catch locality Zastávka u Brna. Samples were obtained from 43 bat carcasses at two different localities in the Czech Republic (Zastávka u Brna, Malá Morávka). Total mercury content varied between 1.76-72.20 µg/g in fur, 0.04-0.14 µg/g in skin, and 0.05-0.20 µg/g in muscle. Total mercury values in the fur of some individuals from Malá Morávka exceeded the recognized toxicity limit. Furthermore, there was a significant difference (p < 0.001) in content of total mercury in fur between localities, and there was a clear effect of age on concentrations in fur, skin, and muscle, the concentrations being significantly correlated (fur and skin rs = 0.783; fur and muscle rs = 0.716; skin and muscle rs = 0.884). These findings confirm the usefulness of fur samples from living bats for biomonitoring mercury burden in the environment.

Mercury distribution in the great cormorant (Phalacrocorax carbo) from the Krogulna ponds and Nysa Klodzka River (Poland).

Mercury belongs among the highly hazardous substances present in the environment and represents a great health risk for many animals including predatory and piscivorous birds. The aim of this study was to assess the total mercury content in the main detoxifying organs (liver and kidney) and in the muscles of adult great cormorants (Phalacrocorax carbo) caught at two localities (the Krogulna ponds and the Nysa Klodzka River) in southern Poland. The aim was to compare the locality with the iron ore mining history (the Krogulna ponds) with a site without such load (the Nysa Klodzka River). The total mercury content in the great cormorants decreases as follows: kidneys > liver > muscle in both monitored localities and significantly differs between the localities. The average mercury content varied from 0.58 ± 0.38 mg/kg (muscle) to 1.39 ± 1.42 mg/kg (kidney) in the cormorant from the Krogulna ponds and from 1.09 ± 0.40 mg/kg (muscle) to 3.12 ± 1.55 mg/kg (kidney) in the cormorant from the Nysa Klodzka River. The accumulation of mercury does not correlate with the mining history as it was higher at the Nysa Klodzka River, but it is probably influenced by the different fish stocks in these two localities (omnivorous fish in the Krogulna ponds versus predatory fish in the Nysa Klodzka River).

Mercury content in fish from drinking-water reservoirs in the Morava River Basin (Czech Republic).

This study focused on the total mercury content in fish from seven drinking-water reservoirs located in the Morava River Basin: Bojkovice, Boskovice, Hubenov, Karolinka, Landstejn, Ludkovice and Nova Rise in the Czech Republic. A total of 308 fish were collected for the analysis. The content of total mercury was measured in the muscle tissue of bream, roach and perch using atomic absorption spectrometry and varied from 0.057±0.009 to 0.440 mg kg-1 in bream, from 0.030±0.005 to 0.393±0.138 mg kg-1 in roach and from 0.092±0.007 to 0.638±0.042 mg kg-1 in perch. The highest total mercury content was found in perch from Landstejn and the lowest was measured in roach from Ludkovice. A positive statistically significant relationship was found between fish weight and total mercury content in fish muscle for almost all species and all sampling sites, except for roach from Ludkovice. A total of 19 samples exceeded the maximum mercury level set by legislation on food contaminants-0.5 mg kg-1 in freshwater fish.

Mercury speciation in fish muscles from major Czech rivers and assessment of health risks.

The aim of this work was to determine the mercury and methylmercury content in muscle tissue of chub (Leuciscus cephalus L.), to assess the health risks of eating the fish and to determine the number of fish meat servings that are suitable for weekly consumption. Total mercury concentrations were determined using a single-purpose atomic absorption spectrophotometer AMA 254. Methylmercury concentrations were determined by gas chromatography. The location where the highest total mercury concentrations in fish muscle tissues were found was the Vltava - Vranany (0.236±0.1001mg/kg(-1)), and the highest methylmercury concentration was found at the Labe - Obríství (0.231±0.1056mg/kg(-1)). The conclusion based on the data ascertained is that the locations from which the lowest number of fish meat servings can be eaten are the Vltava - Vranany and the Labe - Obríství. The results of this study helped evaluate contamination levels of rivers that flow out of the Czech Republic.

Total mercury content in canine hair before and after administration of vaccines containing thiomersal.

OBJECTIVES: Thiomersal is ethylmercury containing compound. It has been used as a preservative in vaccines since the 1930s because it is very effective in preventing bacterial contamination. Ethylmercury penetrates into growing hair in a similar manner as methylmercury. DESIGN: A total of 48 hair samples were collected from vaccinated dogs. Each sample was accompanied with a questionnaire including data on age, gender, vaccinations. Total mercury content in hair, granules and vaccines was determined by the direct method of cold vapours using an AMA 254 (advance mercury analyser; Altec Ltd., Czech Republic). RESULTS: At first we performed two pre-experiments. In first pre-experiment, the highest value of total mercury content was 0.732 mg.kg-1. The content of total mercury ranged from 0.022 to 0.092 mg.kg-1 in the second pre-experiment. The results were not statistically significant in the pre-experiments. In the main experiment the lowest concentration of total mercury in dog's hair was 0.002 mg.kg-1 and the highest value was 0.560 mg.kg-1. The median value of total mercury ranged from 0.023 to 0.033 mg.kg-1. The results were not statistically significant in the main experiment. Total mercury content in vaccines corresponded with the declared quantity. Rather, results showed mercury content to be correlated with the consumption of feed containing fish. CONCLUSIONS: Thiomersal preservative, contained in vaccine, does not increase content of total mercury in canine hair. Our results have shown that content of mercury in hair depends on fish consumption (fish granules, fish treats and fresh fish).

Distribution of mercury in rainbow trout tissues at embryo-larval and juvenile stages.

The aims of the study were to determine total mercury concentrations in "rainbow trout Oncorhynchus mykiss (Walbaum)" at their embryo-larval and juvenile stages and to assess mercury concentration dynamics in individual tissues. Samples of rainbow trout were collected at two-month intervals over a period of 18 months (one stock production cycle) at the Velká Losenice trout farm. Feedstuff samples were collected at the same time and analyzed for mercury concentrations. Tissue mercury concentrations were determined in muscle, liver, and kidneys. Analyses were performed using the AMA 254 atomic absorption spectrophotometer. The lowest mercury concentration was found in 14-day-old embryos (hard roe), and the highest concentrations in muscle tissue, liver, and kidneys at the end of monitoring, that is, in rainbow trout aged 18 months. The amount of mercury in feedstuffs showed an increasing trend and ranged between 0.0126 and 0.0859 mg kg(-1). A significant effect (P < 0.001) of mercury intake on mercury concentrations in muscle tissue, liver, and kidneys was demonstrated. Muscle mercury concentrations in 18-month-old market-ready rainbow trout of 0.128 ± 0.048 mg kg(-1) met the criteria for fish meat hygiene.

The effects of Click 500 SC (terbuthylazine) on common carp Cyprinus carpio under (sub)chronic conditions.

OBJECTIVES: Effects of the herbicide formulation Click 500 SC (terbuthylazine 500 g/l) on common carp Cyprinus carpio were assessed through biometric, biochemical, haematological and antioxidant indices, induction of xenobiotic metabolizing enzymes and histological examination of selected tissues. DESIGN: The fish were exposed to the formulation with terbuthylazine concentrations of 380 ng/l (environmental concentration); 60 µg/l and 550 µg/l for up to 91 days. Haematological indices were assessed using unified methods of haematological examination in fish. Biochemical indices in plasma were measured by biochemical analyzer, ferric reducing ability of plasma (FRAP) and ceruloplasmin activity were determined spectrophotometrically. Concentration of total cytochrome P450, glutathione-S-transferase activity and glutathione content were assessed spectrophotometrically in liver. Activity of liver ethoxyresorufin-O-deethylase (EROD) activity was measured spectrofluorimetrically. Histopathological examination of liver, skin, gills, spleen, cranial and caudal kidney was performed by light microscopy. RESULTS: An increase (p<0.05) was observed in hepatosomatic index and condition factor in fish from the environmental concentration. A decrease (p<0.05) in haemoglobin and mean corpuscular haemoglobin concentration (MCHC) was found in fish treated with terbuthylazine of 550 µg/l. There was a decline in mean corpuscular volume (MCV) and mean corpuscular haemoglobin (MCH) (p<0.05) in terbuthylazine of 60 µg/l and 550 µg/l. Triglycerides (TAG) (p<0.01) were elevated in all pesticide-treated groups. Alanine aminotransferase (ALT) (p<0.01) and phosphorus (p<0.05) decreased in fish exposed to terbuthylazine of 60 µg/l and 550 µg/l, while albumin (p<0.01) rised in the same groups. An elevation in natrium (p<0.05) in terbuthylazine of 550 µg/l and a rise in protein (p<0.01) in the concentrations of 380 ng/l and 550 µg/l were observed. Correlations between several indices were significant. Ceruloplasmin activity and FRAP were augmented (p<0.01) in the highest concentration tested. Examined xenobiotic detoxification systems were not significantly affected by the exposure. Non-specific histopathological changes were found in the gills and skin of the test fish. CONCLUSION: The fish treated with terbuthylazine developed a disorder in several haematological and plasma biochemical indices. The levels of markers of oxidative stress increased in response to the exposure. Examined systems involved in detoxification of xenobiotics did not reflect long-term contact with the herbicide. Detected histological lesions were non-specific. The environmental concentration of terbuthylazin affected biometric indices of the test fish.

Assessment of contamination of the Svitava and Svratka rivers in the Czech Republic using selected biochemical markers.

The aim of the present study is to assess aquatic ecosystem contamination using selected biochemical markers: cytochrome P450, ethoxyresorufin-O-deethylase (EROD), glutathione S-transferase (GST), tripeptide glutathione, vitellogenin, and 11-ketotestosterone in chub (Leuciscus cephalus L.). Seven locations on the Svitava and Svratka rivers (in the Brno conurbation, Czech Republic) were assessed. The results were compared with the levels of the most important inductors of these biomarkers: organic pollutants hexachlorocyclohexane (HCH), hexachlorobenzene (HCB), DDT and its metabolites, polychlorinated biphenyls, and polycyclic aromatic hydrocarbons in bottom sediment, fish muscle, and semipermeable membrane devices (SPMDs) and metals in sediment. The highest levels of pollutants were observed at sites situated downstream from Brno, especially at Modrice and Rajhradice. Significant positive correlations (p < 0.05) were found between EROD activity and HCH concentration in SPMDs, and also between GST and EROD activity with HCB concentration in muscle, after adjusting for age.

1-Hydroxypyrene--a biochemical marker for PAH pollution assessment of aquatic ecosystem.

The aim of the present study was to assess aquatic contamination by polycyclic aromatic hydrocarbons (PAH), using the 1-hydroxypyrene (1-OHP) content in fish bile as a biochemical marker. A total of 71 chub (Leuciscus cephalus L.) were collected from seven locations on the Svitava and Svratka rivers in and around the industrial city of Brno, Czech Republic. The levels of 1-OHP were determined by reverse phase HPLC with fluorescence detection after deconjugation. Normalising the molar concentration of the biliary 1-OHP to the biliary protein content reduced sample variation. The content of 1-OHP was correlated with the PAH level in bottom sediment and semi-permeable membrane devices (SPMD), which was analyzed by a combination of HPLC/FLD and GC/MS methods. The highest mean values of 1-OHP were found in fish caught at the Svratka River at locations Modrice (169.2 ± 99.7 ng · mg(-1) protein) and Rajhradice (152.2 ± 79.7 ng · mg(-1) protein), which are located downstream from Brno. These values were significantly (P < 0.05) higher than those obtained from localities Knínicky (98.4 ± 66.1 ng · mg(-1) protein) and Bílovice nad Svitavou (64.1 ± 31.4 ng · mg(-1) protein). The lowest contents of PAH in sediment and SPMD were found at location Knínicky (1.5 mg · kg(-1) dry mass and 19.4 ng · L(-1), respectively). The highest contents of PAH in sediment and SPMD were found in Rajhradice (26.0 mg · kg(-1) dry mass) and Svitava before junction (65.4 ng · L(-1)), respectively. A Spearman correlation test was applied to determine the relationship between biliary 1-OHP and the sum of PAH in sediment and SPMD. A positive, but no statistically significant correlation was found. The main impact sources of elevated level of PAHs in sites located downstream from Brno are most probably intensive industrial and agricultural activities and domestic waste.

Fish oil and cod liver as safe and healthy food supplements.

OBJECTIVES: The aim of this study is to determine the content of mercury, methylmercury and persistent organic pollutants in the capsules filled with fish oil from marine fish and in the canned cod liver and find out, whether analyse products are safe and suitable for human consumption. METHODS: Total mercury was determined by cold vapour atomic absorption spectroscopy on an AMA-254 (Altec Ltd., Czech Republic) single-purpose mercury analyzer. Methylmercury in the cod liver in the form of chlorid methylmercury was determined by gas chromatography. Seven indicator congeners of PCB (PCB 28, 52, 101, 118, 138, 153, 180), HCH, HCB, DDT and its degradation products DDE and DDD, were determined by gas chromatography-mass spectrometry. RESULTS: In capsules filled with fish oil (n=19) total Hg levels were in the range of 0.013 to 2.03 ng/g. All the capsule oil samples analyzed for MeHg were below the detection limit. The highest concentration of total Hg was found in cod liver - can A (0.223 ng/g). The maximum percentage of MeHg in total Hg concentration was found in a liver sample - can B. The values of alpha-, beta-, gamma-, delta- HCH and of HCB were found below the detection limits in all the capsule samples analyzed. In eight capsules, the presence of highly lipophilic PCB congeners was demonstrated. DDT and its important DDE metabolite were found in ten samples. In canned cod liver the highest concentration of all PCB congeners was demonstrated in can D. The lowest congener concentrations, however, were found in smoked cod liver - can C. DDT with its main metabolite DDE was detected in can C. No additional DDT and DDD persistent pollutants were detected. CONCLUSIONS: The consumption of fish oil in capsules, and canned cod liver is safe and healthy and should be encouraged.

Using human hair as an indicator for exposure to mercury.

OBJECTIVES: Exposure to mercury, a risk factor for neuro-developmental toxicity, was evaluated in the Czech Republic by performing mercury determination using human hair as an indicator. METHODS: Hair samples from Czechs (n=311; 2-66 years old) were analyzed for mercury content. Total mercury was analysed by mercury analyzer AMA 254. RESULTS: The highest total mercury content found in sampled hair was 3.55 microg/g and the lowest content was 0.015 microg/g. No correlation was found between the mercury levels in the sampled hair and the subject's age, gender, and the amount of amalgam fillings. A total of 38 hair samples were analyzed for methylmercury content. CONCLUSION: The results show a positive correlation between the total mercury content in human hair and the consumption of marine and freshwater fish. Hair are a very good indicator of fish consumption.

Fish biochemical markers as a tool for pollution assessment on the Svitava and Svratka rivers, Czech Republic.

OBJECTIVES: The study was designed to assess the pollution of the Svitava and Svratka rivers in and around the industrial city of Brno (Czech Republic) by persistent organic pollutants using selected biochemical markers in chub. DESIGN: Levels of selected biochemical markers were measured in liver and plasma samples of chub. The concentrations of persistent organic pollutants (POPs) were determined in bottom sediment, semi-permeable membrane devices (SPMDs) and muscle samples, and consequently used for correlation with biochemical markers. RESULTS: Significant alterations (p < 0.05) in some biochemical markers were observed and associated with combined exposure to pollutants. The highest levels of pollutants were found at sites situated downstream from Brno. The most widespread changes were identified in the function of phase I detoxifying enzymes. Significant positive correlations were observed in cytochrome P450 content and DDT concentration in the semi-permeable membrane device (p = 0.019, rs = 0.886), and between ethoxyresorufin-O-deethylase activity and content of DDT (p = 0.041, rs = 0.352) and polychlorinated biphenyls (p = 0.034, rs = 0.365) in muscle tissues of indicator fish. CONCLUSION: The results presented in our study indicate the highest contamination of sites situated downstream from Brno, where the intensive industrial and agricultural activities as well as domestic waste and sewage most probably comprise the main impact sources of the enhanced level of pollutants and some biochemical markers in fish.

Effects of subchronic exposure to Successor 600 (pethoxamid 600 g L-1) on common carp Cyprinus carpio.

OBJECTIVES: We investigated the effects of the herbicide preparation Successor 600 on biochemical and haematological indices and the histolopathological parameters of common carp after 28 day exposure. Furthermore, the hepatosomatic index and induction of xenobiotic metabolizing enzymes and biomarkers were assessed. DESIGN: Juvenile fish were exposed to sublethal concentrations of Successor(R) 600 (0.06 mg L-1, 0.22 mg L-1 or 0.60 mg L-1) for 28 days. Haematological indices were assessed using unified methods of haematological examination in fish. Plasma biochemical indices were measured by biochemical analyzer, the vitellogenin concentration in male fish plasma was estimated by direct sandwich ELISA. In hepatopancreas, ethoxyresorufin-O-deethylase (EROD) activity was measured spectrofluorimetrically, concentration of total cytochrome P450 (CYP), glutathion (GSH) content and glutathion-S-transferase (GST) activity were determined spectrophotometrically. Histological changes in samples of hepatopancreas, skin, gills, spleen, head kidney and trunk kidney were examined by light microscopy. RESULTS: Haemoglobin, MCH and MCHC were significantly (p<0.05) reduced in fish treated with Successor(R) 600 of 0.22 and 0.60 mg L-1. LDH was enhanced (p<0.05), in the highest concentration of the preparation. Vitellogenin was detected in all male fish, with no difference between groups. HSI, GSH and GST were elevated (p<0.05), owing to the exposure, whereas CYP and EROD were not affected. Slight histopathological changes were demonstrated in skin, gills and hepatopancreas, with steroid tissue in head kidney samples of 0.60 mg L-1 treated fish. CONCLUSION: Successor 600 affected the haematological profile of the treated fish, while the effects on biochemical indices were less expressed. Male plasma vitellogenin concentrations were not indicative of estrogen disruptive effects after 28 days. The importance of GSH and GST for the metabolisation were demonstrated. In contrast, CYP and EROD were not influenced by any concentration tested. HSI was found to reflex pollution with Successor 600. Histopathological indices caused by the treatment were observed in various tissue samples of the treated fish.

Mercury in human hair as an indicator of the fish consumption.

OBJECTIVES: Mercury and most of its compounds are extremely toxic and should be handled with care. It can be inhaled and absorbed through the skin and mucous membranes. The most toxic forms of mercury are its organic compounds such as dimethylmercury and methylmercury. Fish have a natural tendency to accumulate mercury. Methylmercury is produced by microbial methylation of inorganic mercury in water sediment then it infiltrates the food chain and it consequently accumulates in fish. Fish are the main source of methylmercury in human food. Mercury is transferred into a hair; and this can be than used to monitor the long-term exposure to mercury. The content of mercury in hair depends on the frequency of fish consumption. The aim of our study was to compare mercury content in the hair of children that had various amounts of fish consumption (increased or reduced). DESIGN: Total mercury content in hair was determined by direct method of cold vapors using an AMA 245 analyzer. A total of 174 hair samples from the children (9-17 years old) were analyzed. In this study, the following localities were compared: Neratovice (n=42), Jeseníky (n=44), Prague (n=59) in Czech Republic and Olsztyn in Poland (n=29). Every sample was accompanied with questionnaire about age, gender, regions, amalgam fillings and fish consumption. RESULTS: We did not find a correlation between the content of mercury in hair with age, gender or amalgam fillings. We did find a correlation between fish consumption and the amount of mercury found in the hair samples. CONCLUSION: The amount of mercury in hair increases with more frequent consumption of freshwater and marine fish.

Effects of subchronic metribuzin exposure on common carp (Cyprinus carpio).

OBJECTIVES: Effects of metribuzin on biochemical and haematological indices, induction of specific biomarkers and impacts on biometric parameters of Cyprinus carpio were investigated for subchronic conditions. METHODS: Juvenile fish were exposed to 0.175 mg.L-1 or 1.75 mg.L-1 metribuzin for 28 days. Haematological indices were assessed using unified methods of haematological examination in fish. Biochemical indices were determined by biochemical analyzer, hepatic vitellogenin content was estimated by direct sandwich ELISA. Cytochrome P450 concentration and ethoxyresorufin O-deethylase activity were measured in liver spectrophotometrically and spectrofluorimetrically, respectively. RESULTS: Increased haematocrit and RBC count were found in fish exposed to 1.75 mg.L-1 metribuzin compared to control fish (p<0.05). Other indices contents and biomarkers levels were not significantly changed by either concentration of metribuzin. CONCLUSION: Sublethal metribuzin pollution may have adverse impacts on haematological parameters in common carp.

Biliary 1-hydroxypyrene as a biomarker of exposure to polycyclic aromatic hydrocarbons in fish.

OBJECTIVES: The aim of this study was the assessment of the Svitava and Svratka rivers contamination by polycyclic aromatic hydrocarbons (PAH) using 1-hydroxypyrene (1-OHP) concentrations in fish bile as a biomarker. DESIGN: Levels of 1-OHP were determined by reverse phase HPLC with fluorescence detection. For valid assessment of bile accumulation levels, the 1-OHP concentration was normalized to the biliary protein content. The content of 1-OHP was correlated with the PAH content of river bottom sediments. RESULTS: The highest level of 1-OHP in fish bile (136.1 ng.mg-1 protein) was found at the locality Rajhradice, which is situated downstream of Brno. Also the greatest level of sum of PAHs was found at this locality (17.1 microg.g-1 dry mass). Significant positive correlation (p< or =0.05) between the level of 1-OHP and sum of PAHs in sediment was found only in case of sediments collected in the same month as fish samples (in June). CONCLUSION: Our results document that 1-OHP in fish bile is a suitable biochemical marker for the assessment of aquatic ecosystem contamination by PAHs.

Stress responses in adult cattle due to surgical dehorning using three different types of anaesthesia.

Dehorning adult cattle is a surgical procedure causing distress of varying intensities that can be reflected in behavioural changes and alterations in plasma cortisol levels. Stress responses during the dehorning process were evaluated in 18 Red Pied cows. The cows were divided into 3 groups of six and kept in tie-stall housing. Those in the first group were dehorned under general anaesthesia (GA) induced by intravenous administration of xylazine and ketamine. The second group was dehorned under sedation and local anaesthesia (SLA) induced by intramuscular administration of xylazine and local anaesthesia with lidocaine. The third group was dehorned under local anaesthesia (LA) with lidocaine. Dehorning was performed with a foetotomy wire. Blood samples were taken 0.5 h before dehorning to determine cortisol levels, and, by means of a central venous catheter inserted into the jugular vein, during surgery at 0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 7 and 8 h post-surgery. Concurrently, occurrence of stress behaviours was assessed. Cortisol levels were measured by enzyme immunoassay (EIA). The lowest mean peak levels of plasma cortisol (82.53 +/- 6.04 nmol l(-1)), the most rapid return of plasma cortisol levels to baseline values (1.92 +/- 1.11 h), and the lowest occurrence of stress behaviours (2.38 +/- 5.83%) were noted in the SLA group. The highest mean peak levels plasma cortisol (113.86 +/- 25.65 nmol l(-1)), the slowest return of plasma cortisol levels to baseline values (3.83 +/- 2.18 h) and the most frequent occurrence of stress behaviours (65.48 +/- 28.72%) were observed in the LA group. There were significant differences between the SLA and LA groups in peak plasma cortisol levels (p = 0.011) and in occurrence of stress behaviours (p = 0.003). Sedation induced by intramuscular administration of xylazine in conjunction with local anaesthesia with lidocaine is considered the most suitable method of anaesthesia when dehorning adult cattle. Local anaesthesia with lidocaine alone was least suitable.