Hemerocallis as bioindicator of fluoride pollution in tropical countries

In the frame of an active monitoring study at the Serra do Mar near the industrial pool of Cubatão, Brazil,Hemerocallis was tested for its suitability as bioindicator of airborne fluoride pollution. In a screening experiment with various cultivars comparison of susceptibility, correlation of visible injury and foliar fluoride concentration as well as comparison with exposure ofGladiolus gave best results for theHemerocallis cultivar ‘Red Moon’. When exposed simultaneously foliar fluoride accumulation ofHemerocallis showed a highly significant linear correlation with fluoride content ofLolium multiflorum, the ‘standardized grass culture’ andGladiolus, well-known bioindicator species for fluoride impact. With respect to the extent of foliar injuryHemerocallis was less sensitive thanGladiolus, in terms of accumulation capacity it ranged betweenLolium andGladiolus. In general, the results of the biomonitoring study proved that the Atlantic Forest vegetation in a valley downwind from the Cubatão fertilizer industries is still suffering from severe fluoride pollution.     

Assessing water quality in a tropical lake using biomarkers in zebrafish embryos: developmental toxicity and stress protein responses

In order to achieve a more substantial appraisal of lake water quality, the assessment must not be based only on chemical measurements and analyses of the water itself, but even more so on the impact of existing conditions on aquatic biota. This is possible with the use of biotests or biomarkers, e.g. investigations of the developmental parameters (96-h embryotoxicity evaluate) or of the induction of heat shock proteins (proteotoxicity evaluate). To evaluate the suitability of these tests for environmental screening, fertilized zebrafish eggs were exposed to water samples collected from five sites of varying levels of stress from Laguna Lake, Philippines. Reconstituted water was used as laboratory control while water samples from a highly polluted freshwater source was used as positive control. Developmental parameters were noted and described within 48 and 96 h of exposure. Dilution experiments of the positive control were also done to further assess and compare toxicity potentials of Laguna Lake waters with those originating from a polluted freshwater. After the 96-h exposure, the levels of stress proteins (hsp 70) were determined in embryos from all exposure groups. Results showed 100% mortality in embryos exposed to undiluted positive control (PC) within only 12 h. Increasing dilution levels, however, resulted in lower mortality and lower abnormality rates. No detectable developmental differences were noted among embryos exposed to either the laboratory control or Laguna Lake waters at the end of 96 h, regardless of the source. Very high survival rates and high hatching success rates were observed in embryos exposed to lake waters as well as laboratory control, and the data did not differ significantly among the groups. Likewise, no significant malformations were noted among all developing embryos throughout the exposure period. However, the levels of heat shock proteins in the two sites located closest to Manila, the Philippine capital (Northern West Bay and Central West Bay), showed a pronounced elevation relative to the control, indicating that these stress proteins protect the embryos from the detrimental effects of pollutants in the water. Based on the 96-h early life stage (ELS) test, the water quality of the lake is good for fishery propagation despite the current levels of pollutants in the water. This finding is in accordance with the Class C status (i.e. suitable for fish growth and propagation) as given to the lake by the local environmental agency. On the other hand, data on proteotoxicity showed that the fish are under stress, presumably deriving from pollutants. This calls for a continuous monitoring and improvement of the lake water. The present study indicates that the two biomarker methods are very easy to use, practical, rapid, and sensitive for assessing water quality in a tropical lake and recommends for their incorporation into the future monitoring program of Laguna Lake.     

Histopathology alterations and lipid peroxidation as biomarkers of hydrocarbon-induced stress in earthworm, Eudrilus eugeniae

This study investigated the biomarker responses of the earthworm, Eudrilus eugeniae, exposed to sublethal concentrations of benzene, toluene and xylene (BTX) for a time span of 28 days. Lipid peroxidation (LPO) and histopathological alterations were examined. Toxicological evaluations of BTX were carried out against E. eugeniae. On the basis of the 96-h LC₅₀ value, xylene (1.212 mg/kg) was found to be the most toxic followed by toluene (1.335 mg/kg) and benzene (1.896 mg/kg) was the least toxic. The exposure of earthworms to sublethal concentrations (1/10th and 1/100th of 96 h LC₅₀) of BTX premixed with the substrate (loamy and humus soil) induced pathological changes in the clitella such as severe lesion, necrosis and dark brown pigments. The result of the lipid peroxidation assay showed a significant increase in oxidative damage with LPO values ranging from 2.58–7.8 nM/g in exposed animals when compared to 0.07 nM/g in the control group. The findings from this study suggest the use of LPO and histopathology as useful biomarkers of exposure for early detection of petroleum related stress in terrestrial ecosystems     

Induction of sperm impairments in mice as a sensitive biomarker of arsenic toxicity

The ubiquitous presence of arsenic (a toxic metalloid) in our environment, particularly in our drinking water, is a serious health hazard of global concern. The present work deals with the assessment of arsenic toxicity through the analysis of induced sperm impairments in sperm head morphology and sperm count in mice at low exposures compared to the magnitude of response at high exposure levels. The animals were exposed to four doses of arsenic, ranging from lowest dose of 0.3 μg kg^?1 day^?1 (the human reference dose) to higher dose of 30 μg kg^?1 day^?1 for 15 consecutive days. The epididymal sperms were harvested after one spermatogenic cycle on the 36th day and were scored for the presence of any abnormality in their head morphology as well as changes in their count. Exposure to arsenic significantly induced, in a dose-dependent manner, increases in the frequency of sperms with abnormal head morphology from 5.12 % in control to 9.23 % in lowest dose group and 23.02 % in highest dose group. In contrast, the mean sperm counts in the epididymal wash were decreased from 6.05 million per milliliter in the control to 4.95 million per milliliter in the lowest dose group and 3.07 million in the highest dose group. The analysis of sperm impairments in mice was, therefore, found to be a highly sensitive assay to assess arsenic toxicity, exhibiting a marked male reprotoxic effect of arsenic even at its low exposure levels including the human reference dose.     

Naphthalene and its biomarkers as measures of occupational exposure to polycyclic aromatic hydrocarbons

Polycyclic aromatic hydrocarbons (PAH) include compounds with two or more fused benzene rings, many of which are carcinogens. Industrial sources produce hundreds of PAH, notably in the coke- and aluminium-producing industries. Because PAH are distributed at varying levels between gaseous and particulate phases, exposure assessment has been problematic. Here, we recommend that occupational exposures to naphthalene be considered as a potential surrogate for occupational PAH exposure for three reasons. Naphthalene is usually the most abundant PAH in a given workplace; naphthalene is present almost entirely in the gaseous phase and is, therefore, easily measured; and naphthalene offers several useful biomarkers, including the urinary metabolites 1- and 2-hydroxynaphthalene. These biomarkers can be used to evaluate total-body exposure to PAH, in much the same way that 1-hydroxypyrene has been applied. Using data from published sources, we show that log-transformed airborne levels of naphthalene are highly correlated with those of total PAH (minus naphthalene) in several industries (creosote impregnation: Pearson r= 0.815, coke production: r= 0.917, iron foundry: r= 0.854, aluminium production: r= 0.933). Furthermore, the slopes of the log-log regressions are close to one indicating that naphthalene levels are proportional to those of total PAH in those industries. We also demonstrate that log-transformed urinary levels of the hydroxynaphthalenes are highly correlated with those of 1-hydroxypyrene among coke oven workers and controls (r= 0.857 and 0.876), again with slopes of log-log regressions close to one. These results support the conjecture that naphthalene is a useful metric for occupational PAH exposure. Since naphthalene has also been shown to be a respiratory carcinogen in several animal studies, it is also argued that naphthalene exposures should be monitored per se in industries with high levels of PAH.     

Protein adducts as biomarkers of exposure to aromatic diisocyanates in workers manufacturing polyurethane (PUR) foam

This work was undertaken to investigate the usefulness of diisocyanate-related protein adducts in blood samples as biomarkers of occupational exposure to toluene diisocyanate (TDI; 2,4- and 2,6-isomers) and 4,4'-methylenediphenyl diisocyanate (MDI). Quantification of adducts as toluene diamines (TDAs) and methylenedianiline (MDA) was performed on perfluoroacylated derivatives by gas chromatography-mass spectrometry (GC-MS/MS) in negative chemical ionisation mode. TDI-derived adducts were found in 77% of plasma and in 59% of globin samples from exposed workers manufacturing flexible polyurethane foam. The plasma levels ranged from 0.003 to 0.58 nmol mL(-1) and those in globin from 0.012 to 0.33 nmol g(-1). The 2,6-isomer amounted to about two-thirds of the sum concentration of TDA isomers. MDI-derived adducts were detected in 3.5% of plasma and in 7% of globin samples from exposed workers manufacturing rigid polyurethane foam. A good correlation was found between the sum of TDA isomers in urine and that in plasma. The relationship between globin adducts and urinary metabolites was ambiguous. Monitoring TDI-derived TDA in plasma thus appears to be an appropriate method for assessing occupational exposure. Contrary to TDI exposure, adducts in plasma or globin were not useful in assessing workers' exposure to MDI. An important outcome of the study was that no amine-related adducts were detected in globin samples from TDI- or MDI-exposed workers, alleviating concerns that TDI or MDI might pose a carcinogenic hazard. Further studies are nevertheless required to judge whether diisocyanates per se could be such a hazard.     

Sea-urchin (Paracentrotus lividus) glutathione S-transferases and cholinesterase activities as biomarkers of environmental contamination

Activities of glutathione S-transferases (GST) and cholinesterase (ChE) from Paracentrotus lividus were investigated as possible biomarkers of environmental contamination in the coastal zone. In the first phase of the study, the activity of both enzymes was determined in various tissues in order to select the most appropriate ones to be used in the following assays. In the second phase, the ChEs present in ambulacra were characterized using different substrates and selective inhibitors. In the next phase, laboratory bioassays were performed with dilutions of water-accommodated fraction of #4 fuel-oil (WAF) and benzo[a]pyrene (BaP) to determine the response of those enzymes to these pollutants and, finally, the activity of both enzymes was determined during a year in indigenous specimens from six sites on the Northwest coast of Portugal, with different pollution levels, to determine basal values and seasonal variations of ChE and GST activities. Among the several tissues tested, ambulacra and the anterior portion of the intestine were selected for ChE and GST assays, respectively. The determination of ChE in ambulacra tissue may be performed in a non-destructive way. Ambulacra ChE hydrolysed acetylthiocholine preferentially to propionylthiocholine and butyrylthiocholine and, inhibition by excess of substrate was observed. Enzymatic activity was almost fully inhibited by eserine sulfate (>98%) at concentrations equal or higher than 6.25 microM. Sensitivity to both BW284C51 (reaching 98% at 200 microM) and iso-OMPA (73% at 8 mM) was found. In laboratory bioassays, GSTs activity was inhibited by WAF and induced by BaP, whereas ChE activity was not affected by any of these environmental contaminants. Seasonal variations in enzymatic activities were found. For example, in a reference site, ChE values changed from 52.2 +/- 9.3 U mg(-1) protein in autumn to 71.8 +/- 13.3 U mg(-1) protein in spring, while GST activity changed from 129.9 +/- 29.8 U mg(-1) protein in winter to 279.0 +/- 48.0 U mg(-1) protein in autumn. Sea-urchins from reference sites presented significantly higher values of both ChE and GST than animals from contaminated sites in all seasons. In conclusion, the results of this study indicate that (i) ambulacra and the anterior portion of intestine are the most suitable tissues to measure ChE and GST activities, respectively; (ii) only one form of ChE seems to be present in ambulacra, showing properties of both typical acetylcholinesterase (AChE) and pseudocholinesterase (PChE); (iii)P. lividus GST is sensitive to both WAF and BaP even after acute exposures while ChE is not, and (iv) in spite of the significant seasonal variations observed in both enzymes in the field, P. lividus ChE and GST were capable of discriminate sites with different contamination levels and, thus, they are suitable for use as biomarkers in biomonitoring studies in the coastal zone.     

Occurrence, source apportionment and toxicity assessment of polycyclic aromatic hydrocarbons in surface sediments of Chaohu, one of the most polluted lakes in China

In order to evaluate the effect of local anthropogenic activities on Chaohu Lake, one of the most eutrophicated lakes in China, surface sediments have been collected from the whole lake with 0.05 × 0.05 degree latitude/longitude resolution and in the estuaries of three main inflowing rivers. The concentrations of the 28 polycyclic aromatic hydrocarbons (PAHs) determined were in a range 82.4-13,000 ng g(-1) with an average value of 1670 ng g(-1) dry weight for total 28 PAHs (referred to as Σ(28)PAH). Amongst the 28 PAHs, 16 are listed as high priority PAHs by the USEPA and they were in the range of 60.8-10,200 ng g(-1) with an average value of 1230 ng g(-1) for the total of them (referred to as Σ(16)PAH); 7 are known as carcinogenic PAHs and their levels ranged from 34.2 to 6400 ng g(-1) with an average of 815 ng g(-1) in total (referred to as Σ(7)PAH). Chaohu Lake was considered significantly polluted by PAHs through the comparison with the PAH burdens in fresh-water lakes both in China and worldwide. Toxic units (TUs) evaluation showed some sampling locations possibly were over the median lethal level for benthic invertebrate. The highest PAH concentrations were found in sediments from the Nanfei River estuary, suggesting the major contributor of PAHs contamination to the lake. The PAHs with four and five rings were found to be dominant among the PAHs detected in all of the sediment samples, and perylene was the most abundant. Σ(16)PAH had a good correlation with those PAHs from pyrogenic sources, such as anthracene and phenanthrene, but a poor correlation with perylene. The results demonstrated that the environmental behavior of PAHs from pyrogenic sources is significantly different to that of perylene from diagenetic sources. The PAHs in sediments were mainly from traffic-related emission by qualitatively assessing with the diagnostic ratios of PAH isomers, and the ratios for low molecular weight PAHs were strongly altered during their transport.     

Oxidative DNA damage levels and catalase activity in the clam Ruditapes decussatus as pollution biomarkers of Tunisian marine environment

Levels of the oxidative DNA damage 7, 8-dihydro-8-oxo-2′-deoxyguanosine (8-oxodG) and catalase (CAT) activity were measured in the digestive gland and gills of clams Ruditapes decussatus, related to the presence of pollutants along Tunisian marine environment. Increased levels of CAT were observed in tissues of clams from all the sites studied, compared to control values, and elevated 8-oxodG levels were observed at specific sites. Results obtained in this work indicate that the measurement of 8-oxodG levels and CAT activity in tissues of R. decussatus is promising in pollution monitoring studies of the Tunisian marine environment.     

Cytochrome P450 system as potential biomarkers of certain toxicants: comparison between plant and animal models

In present study, we measured the activities of some selected cytochrome P450 isozymes like ethoxyresorufin-o-deethylase (EROD), pentoxyresorufin-o-deethylase (PROD), and N-nitrosodimethylamine demethylase (NDMA-d) treating experimental rats with different environmental toxicants, namely trichloroethylene (TCE), heavy metal mixture, and ethyl alcohol individually or in combination and by exposing the Allium cepa bulbs to increasing concentration of TCE for 48 h. In animal system, NDMA-d activity displayed a remarkable amplification by 4.2-fold in the liver of alcohol ingested rats compared with control animals. TCE intake also resulted in a marked increase of around 3.8-fold and 1.2-fold in rats’ NDMA-d activity in the liver and kidney, respectively. In onion bulbs, an amazing rise in the activity of test enzymes was recorded in a dose-dependent manner. Among the said enzymes, PROD showed maximum increase up to the extent of 22-fold in comparison with control at 20 ppm of TCE exposure, whereas NDMA-d and EROD showed 11- and 9-fold increase in enzymatic activity, respectively, compared with the control. Based on this study, we conclude and advocate that all the selected isozymes of CYP450 system, viz. PROD, NDMA-d, and EROD can act as potent biomarkers in plant system for assessing the TCE pollution.     

Lipid peroxidation and antioxidant defense enzymes in Clarias gariepinus as useful biomarkers for monitoring exposure to polycyclic aromatic hydrocarbons

The toxicological evaluations of crude oil, xylene, toluene and benzene were carried out against juvenile stages of Clarias gariepinus. On the basis of LC(50) value, benzene (0.017 ml/l) was found to be the most toxic followed by xylene (0.086 ml/l), toluene (0.398 ml/l) and crude oil (2.219 ml/l) was the least toxic. The results of the lipid peroxidation assay showed that the level of malonaldehyde (MDA) in liver and gills of fish exposed to all the test chemicals increased significantly (P < 0.05) when compared to control. Measurement of activities of antioxidant enzymes, superoxide dismutase (SOD), catalase (CAT) and glutathione-S-transferase (GST) in liver and gill of fishes exposed to sublethal concentration of the test chemicals over a 28-day period was found to decrease significantly (P < 0.05) when compared to control. The observed reduction in the activities of antioxidant defense enzymes such as SOD, CAT and GST in conjunction with an increase in MDA levels in the gill and liver tissues of test animals exposed to aromatic hydrocarbon compounds were recommended as a good battery of biomarkers for early detection of pollution during biomonitoring programmes.     

Artemia salina as test organism for assessment of acute toxicity of leachate water from landfills

Artemia salina has, for the first time, been used as test organism for acute toxicity of leachate water from three landfills (the municipal landfills at Kristianstad, Sweden and Siauliai, Lithuania, and an industrial landfill at Stena fragmenting AB, Halmstad, as well as for leachate from Kristianstad treated in different ways in a pilot plan). Artemia can tolerate the high concentrations of chloride ions found in such waters. Large differences in toxicities were found, the leachate from Siauliai being the most toxic one. To increase the selectivity in the measurements, a fractionation was done by using ion exchange to separate ammonium/ammonia and metal ions from the leachate, and activated carbon adsorbents for organic pollutants. The influence of some metals and phenol compounds on the toxicity was investigated separately. It was found that most of the toxicity emanated from the ammonium/ammonia components in the leachate. However, there was also a significant contribution from organic pollutants, other than phenol compounds, since separate experiments had in this latter case indicated negligible impact. The concentrations of metals were at a level, shown by separate experiments, where only small contribution to the toxicity could be expected.     

Seasonal variation of Sarpa salpa fish toxicity, as related to phytoplankton consumption, accumulation of heavy metals, lipids peroxidation level in fish tissues and toxicity upon mice

The aim of this work was to investigate for Sarpa salpa the seasonal trend in the food sources, heavy metals bioaccumulation and the oxidative stress in the organs. In addition, the toxicity was assessed by mouse bioassay of extract of the fish's organs collected in autumn, the peak of occurrence of hallucinatory syndrome. The toxicity was further studied for compounds present in epiphyte collected from the sea at the end of spring and in summer that are digested by the S. salpa in these seasons. We observed a higher lipid peroxydation in different tissues of S. salpa compared to the control fish Diplodus annularis. Furthermore, heavy metals accumulation in organs of these fish showed a significant variation between the two species (P?<?0.05). The lethal dose (LD50%) determined for crude ciguatoxin (neurotoxins) extracts of viscera, liver, brain and muscle of S. salpa were as follows: 1.217, 2.195, 14.395, 18.645 g/kg mouse, respectively. We noticed a significant correlation (P?<?0.05) between the total amount of toxic dinoflagellates and the level of TBARS in the liver, the brain and the muscle, this for all seasons and all sizes. Moreover, the cytotoxic effect observed for epiphytes extract confirms the transfer of toxins originating from toxic dinoflagellates, which live as epiphytes on P. oceanica leaves, to the fish by grazing. Our work indicates that, toxic phytoplanktons and heavy metals accumulation are responsible for the increase of oxidative stress in the organs of S. salpa. Hence, the edible part of S. salpa, especially the viscera and liver, can cause a threat to human health, and consumption should, for this reason, be dissuaded.     

Fish scales as indicators of wastewater toxicity from an international water channel Tung Dhab drain

The effect of wastewater exposure on scales and chromatophores of freshwater fish Channa punctatus was studied using wastewater dilutions (60–100%) from an international water channel Tung Dhab drain at an interval of 15 and 30 days. The exposed fish showed significant alterations such as uprooted and damaged lepidonts and dispersal of chromatophores. These observations strongly suggest that fish scales can be successfully employed as indicators of wastewater pollution.     

发光菌监测浑河(抚顺段)底泥的生物毒性

利用发光茵对浑河(抚顺段)底泥进行了生物毒性测试.结果表明,浑河(抚顺段)主断面底泥均为低毒性,七条支流中以将军河底泥毒性最大,为重毒级,其它支流为低毒级.     

Theoretical aspects of fluoride air contaminant formation in aluminium smelter potrooms

The amount of particulate fluorides evolved from aluminium electrolysis cells is not entirely accounted for by the fluorides entrained in the anode gas. The largest additional source of particulate fluoride formation is by direct evaporation of fluorides into the anode gas stream and subsequent condensation on the drops of electrolyte generated in the process of bubble burst. A theoretical model was used for the calculation of the main physical parameters responsible for the formation of particle nuclei when the hot anode-gas is mixed with ambient air. The results of these calculations are in agreement with experimental observations reported in the literature. In particular, the size distribution, composition and morphology of the nano-particles support the theory of a vapour condensation mechanism under conditions of extreme supersaturation, but further studies are necessary.     

The assessment of biomarkers in sentinel cattle for monitoring vanadium exposure

Various potential biomarkers were sampled for vanadium every 3-4 months from Bos indicus beef cattle farmed extensively immediately adjacent (high exposure (HE) group) and two km away (low exposure (LE) group) from a vanadium processing plant, respectively. Vanadium intake (mg vanadium kg(-1) bwt d(-1)) was modelled using environmental and physiological data as inputs. The vanadium intake ranged from 0.57 to 5.44 mg vanadium kg(-1) bwt d(-1) in the HE group and 0.41 to 2.61 mg vanadium kg(-1) bwt d(-1) in the LE group over a five-year period of monitoring. Samples collected from live sentinel animals over the five-year period included caudal coccygeal vertebrae, tail-switch hair, milk, urine, faeces, rib-bone biopsies and a wide range of blood clinical pathology and haematological parameters. The data was analysed for differences in response between the HE and LE groups. Where differences were found, a linear mixed-effects regression model was fitted to model the relationship between the exposure dose and the response variable. The model included the effects of age, duration of exposure and response, and allowed the prediction of the exposure dose given these inputs. Moreover, forty-two adult cattle were slaughtered over the five years. A wide range of tissue samples, rumen content and whole blood were taken from the cattle at slaughter for vanadium determination. In live animals, a difference in response was found between the HE group and LE group with respect to serum albumin (n = 36), monocyte (n = 36) and thrombocyte (n = 36) counts, and hair (n = 2) and faeces (n = 34) vanadium concentrations. No difference in vanadium concentrations could be shown for urine (n = 36), the traditional occupational health biomarker. Regression models are described for serum albumin, monocyte counts, faeces and hair, which showed the most promise as biomarkers. Average concentrations of vanadium in the tissues of slaughtered cattle ranged from 0.08 to 2.94 mg kg(-1) (wet-weight basis) and rumen content contained 16.67 mg kg(-1). Significant correlations were found between the exposure dose (end-dose) just prior to slaughter and the concentrations of vanadium in the coccygeal vertebrae, liver, diaphragm and rib-bone in descending order of magnitude. Other tissues showed poor correlation to the end-dose. Tissue levels of vanadium in healthy cattle include a much wider range than is currently reflected in the literature. The best tissue from slaughter animals for assessing chronic vanadium exposure is probably the liver.