Genotoxicity of two novel pesticides for the earthworm, Eisenia fetida

In this paper, several studies were conducted to evaluate the genotoxicity of two pesticides, Imidacloprid and RH-5849, for earthworm (Eisenia fetida). Earthworms were exposed in different exposure systems to evaluate their acute toxicity and the genotoxicity of the two pesticides was evaluated by using the method of sperm deformity assessment, micronucleus test of root tip cells in Vicia faba, a mouse bone-marrow micronucleus test, and comet assay. LC(50) (interpolated concentration at which 50% mortality of test population occurs) for earthworms varied in different exposure systems. The results indicated that Imidacloprid was consistently more toxic than RH-5849 in all exposure systems. In this study, sperm deformity test was used to detect the potential adverse influences of pesticides on the reproduction of earthworms. The results demonstrated that significant induction of sperm deformity (p<0.01) and a dose-effect relationship displayed at Imidacloprid concentrations higher than 0.5 mg/kg dry soil. However, the sperm deformity frequency of groups exposed to RH-5849 did not show significant difference (p>0.05) from the control until the dose reached 100 mg/kg dry soil. The results of the V. faba micronucleus tests showed that micronuclei frequency of the exposed group did not show significant difference (p>0.05) from the control until the concentration of Imidacloprid and RH-5849 reached 100 mg/ml. The results of the mouse bone-marrow micronuclei test also indicate that two pesticides did not show significant effects (p>0.05) on the micronuclei frequency in mice bone-marrow cells until the dose reached 100 mg/kg for Imidacloprid and 300 mg/kg for RH-5849 (2/3 LD(50)). Although no genotoxicity was detected by using the micronucleus tests, the results of the comet assay showed that the two pesticides induce significant DNA damage (p<0.01) in earthworms and dose-effect relationships were displayed. The 'earthworm comet assay' is a rapid and sensitive way to screen chemicals or terrestrial environments for their DNA-damaging properties.     

Modulation of hepatic and intestinal glutathione S-transferases and other antioxidant enzymes by dietary lipids in streptozotocin diabetic rats

Antioxidant enzymes in liver and small intestine were investigated using control and streptozotocin diabetic rats fed diets with 5% olive, sunflower or fish oil for five weeks. In liver, Glutathione Peroxidase and Superoxide Dismutase decreased and in intestine Glutathione-S-transferase (GST) increased by diabetes. In isolated jejunum and ileum, this increase in GST activity was due to an increase in GST-alpha and -mu isoenzymes in jejunum and GST-alpha, mu and -pi in ileum. Since GST plays an important role in protecting tissues from oxidative damage, our results highlight the role of the intestine against free radicals in physiological or pathological situations.     

Decreased blood level of β2-microglobulin in the employees of a factory which produced polychlorinated biphenyls

Under the project of evaluating the health status of the employees of CHEMKO factory (East Slovakia) which produced PCBs between 1955 and 1985, the level of β₂-microglobulin (β₂-m) was measured in the serum of 242 subjects from CHEMKO (Group A) and two control groups from much less polluted areas: 1. 277 females from a small mountainous village (Group B), 2. 179 adults from the area of a large city of Ko ice (Group C). The level of thymidine kinase (TK) was measured in the groups A and B only. In addition, age-matched groups of 155 women each from all areas were evaluated. In both the whole group and the age-matched group from CHEMKO the level of β₂-m was significantly lower (P<0.001) than that in the respective control groups, while no difference was found in the level of TK. In conclusion, it is suggested that the decrease of β₂-m in CHEMKO employees might be related to the immunotoxic effects of organochlorines.     

Effects of ozone on yield, growth, and root starch concentrations of two alfalfa (Medicago sativ A L.) cultivars

Ozone (O3) is considered to be a major air pollutant that affects the yield of several sensitive crop species. Its concentration may reach phytotoxic levels several times during the growing season in Eastern Canada. This study was initiated to evaluate the O3 effects on alfalfa, a major crop species. The objective was to compare the yield and growth parameters of the main alfalfa cultivar used in Québec, Apica, to a cultivar more tolerant to O3, Team. Effects on root starch concentrations were also examined as this parameter is an important indicator of alfalfa perennity. The results obtained have shown that the forage yield of Apica was more reduced by O3 during two growing seasons than the yield of Team. For O3 concentrations of 20 to 40 nl liter(-1), yield reductions were 14-26% for Apica and 0-20% for Team. Whereas Apica could be considered more susceptible to O3 than Team, the latter has shown contrasting responses from year to year. This fact suggests that the mechanisms involved in O3-tolerance could be modulated by environmental conditions. At low O3 levels, Apica has shown reduced root growth in terms of dry matter and length. However, contrary to the current hypothesis that O3 would affect more root than shoot growth, we were unable to show a consistent alteration of the biomass allocation between the two. Ozone seems to reduce globally the growth of the whole plants. The greater O3-tolerance of Team could partly be associated to its capacity to maintain more leaves, to delay their senescence, or to keep a larger leaf:stem ratio under increasing levels of O3. At the end of the two growing seasons, the amount of starch reserves stored below ground was shown to be reduced by the current O3 levels. This reduction was mainly associated with a decrease in root biomass under O3 stress. This result support the hypothesis that O3 may accelerate alfalfa decline under field conditions.     

Assessing the efficacy of dredged materials from lake panasoffkee,Florida: Implication to environment and agriculture

BACKGROUND, AIMS AND SCOPE: Current dredged material disposal alternatives have several limitations. Options for dealing with dredged materials include leaving them alone, capping them with clean sediments, placing them in confined facilities, disposing of them at upland sites, treating them chemically, or using them for wetlands creation or other beneficial uses The ability to reuse lake-dredge materials (LDM) for agricultural purposes is important because it reduces the need for offshore disposal and provides an alternative to disposal of the materials in landfills. Often these materials can be obtained at little or no cost to the farmers or landowners. Thus, forage production offers an alternative to waste management since nutrients in the LDM are recycled into crops that are not directly consumed by humans. The objective of this study (Part 2) were to: (1) assess dredge materials from Lake Panasoffkee, Florida as a soil amendment to establish bahiagrass (BG) in a subtropical beef cattle pasture in Sumter County, Florida; and (2) determine the effect of LDM application on the crude protein (CP) and nutrient uptake of BG. This series of two papers aims at providing assessment of the efficacy of lake-dredged materials especially its implication to environment (soil quality, Part 1) and agriculture (forage quality and pasture establishment, Part 2). METHODS: The experimental treatments that were evaluated consisted of different ratios of natural soil (NS) to LDM: LDM0 (100% NS:0% LDM); LDM25 (75% NS:25% LDM); LDM50 (50% NS:50% LDM); LDM75 (25% NS:75% LDM); and LDM100 (0% NS:100% LDM). Bahiagrass plots at its early establishment were cut to a 5-cm stubble height on Julian days 112 and harvested to the same stubble height on Julian days 238 and on Julian days 546 following the double-ring method. Field layout was based on the principle of a completely randomized block design with four replications. Plant samples harvested at 546 Julian days were ground to pass through a 1-mm mesh screen in a Wiley mill. Ground forage was analyzed for crude protein. Ground forage samples were also analyzed for tissue P, K, Ca, Mg, Mn, Cu, Fe, Al, and Mo concentrations using an ICP spectroscopy. The effects of dredged materials addition on forage yield and on crude protein and nutrient uptake that were taken at 546 Julian days were analyzed statistically following the PROC ANOVA procedures. RESULTS AND DISCUSSION: Part 1 of this study demonstrated that the heavy and trace metal contents of LDM were below the probable effect levels and threshold effect levels. As such, the agricultural or livestock industry could utilize these LDM to produce forages. Resuits showed consistently and significantly (p < 0.001) higher BG biomass production and CP from plots amended with LDM than those of BG planted on plots with 0% LDM. Forage yield of BG during its establishment increased linearly (Forage Yield = 1724.3 + 25.64*LDM; R2 = 0.83; p < or = 0.0001) with increasing rates of LDM application. The CP of BG also varied significantly with varying levels of LDM applications. The tissues of BG with 100% LDM had the greatest CP content while the lowest CP content was from the control plots (LDM0). The CP of BG increased linearly with increasing rates of LDM application. The crude protein response to BG application can be described by a linear equation: Crude Protein = 10.38 + 0.052*LDM; R2 = 0.85 p < or = 0.0001. Addition of LDM had increased the levels of Ca by about 1811% when compared with the level of soil Ca among plots with no LDM application. Liming the field could have some direct and indirect effects on the chemical status of the soils. The physiological functions performed by Ca in plants are not clearly defined, but it has been suggested that Ca favors the formation of and increases the protein content of mitochondria. CONCLUSIONS: Beneficial uses of dredged materials from LP, Florida are both economical and environmental. Often these materials can be obtained at little or no cost to the farmers or landowners. Results showed that dredged materials can be used as soil amendments (lime and fertilizer) for early establishment of BG in beef cattle pastures. Environmentally, dredging of sediments that are rich in CaCO3 should restore the 19.4-sq km LP by removing natural sediments from the lake bottom to improve the fishery, water quality, and navigation of the lake. The nutritional uptake of BG grown in unfertile sandy soils of Sumter County was enhanced significantly (p < or = 0.001) by LDM addition. Uptake of TKN, TP, K, Ca, and Mg were remarkably increased as a result of LDM. RECOMMENDATION AND OUTLOOK: Land application of LDM from LP may not only provide substantial benefits that will enhance the environment, community, and society in south Florida, but also in other parts of the world especially those areas with forage-based beef cattle pastures and similar climatic conditions. The heavy and trace metal contents of these materials were below the PEL and TEL (see Part 1). As such, the agricultural or livestock industry could utilize these LDM to produce forages. LDM should be regarded as a beneficial resource, as a part of the ecological system. Although our results have demonstrated the favorable and beneficial effects of added LDM on the early establishment of BG in pasture fields., further studies are still needed not only in pastures of south Florida, but also in other areas with subtropical or tropical climatic conditions to determine whether the environmental and ecological implications of LDM application are satisfied over the longer term.     

Toxic Effects of Crude Oil Combined with Oil Cleaner Simple Green on Yolk-sac Larvae and Adult Rainbow Trout Oncorhynchus mykiss (4 pp)

Background, Goal and Scope Cleaner CRYSTAL Simple Green (SG) was used for the cleanup of the oil spill in the Baltic Sea near Lithuania in 2001. No scientific data are available on the effects and consequences of its application for local aquatic life. The aim of this study was to determine and compare sublethal effects of a) solution SG; b) crude oil alone; c) SG in combination with oil on rainbow trout Oncorhynchus mykiss at different stages of its development in laboratory conditions.Methods Laboratory studies were performed on adult rainbow trout (4-day duration) and on yolk-sac larvae (25-day duration) evaluating their biological parameters. Concentrations of water-soluble and thin-dispersed fractions of petroleum hydrocarbons were measured using gas chromatography.Results and Discussion SG solution (0.5 mg/l) did not affect the survival of larvae and adult fish, and no significant changes were determined in respiratory parameters of the exposed larvae and adult fish. The most expressed alterations were found in morphological parameters (a decrease in the average body mass) of larvae and in haematological indices (a decrease in the leukocyte count) of adult fish at the end of the tests. Crude oil (1610 mg/l) did not affect the survival of adult fish during the 4-day exposure. An increase in larvae mortality rate (~ 36%) was recorded at the end of the tests. A significant decrease in the average body mass and heart rate of larvae as well as in gill ventilation frequency of larvae and adult fish were determined. SG combined with oil induced an increase in larval mortality &#61566; 46% of individuals died at the end of the tests. No mortality was recorded in adult fish. The average body mass and heart rate of larvae were significantly decreased. Marked changes were also found in respiratory parameters (gill ventilation frequency of larvae and adult fish significantly decreased, while “coughing” rate increased). A 1-day, 2-day exposure of fish to SG combined with oil induced a significant decrease in the leukocyte count of adult fish, which was also determined at the end of the tests. The augmentation of adverse impact could be explained by the data obtained from our studies. When SG was added into dilution water with crude oil the concentration of petroleum hydrocarbons in the mixture increased 3 ~ 4.5 times after 24 h and 96 h, respectively. Conclusions The comparative study of the effects of crude oil alone, SG and SG combined with oil showed that their toxic effects on fish differed. Oil combined with SG was found to be more toxic to fish (larvae and adults) than SG alone and oil alone. Fish at early stages of development (yolk-sac larvae) were more sensitive to the effects of the compounds studied than adults.Recommendations and Outlook To diminish the negative impact of oil spill cleanup using chemicals on aquatic ecosystems, it is recommended to carry out more comprehensive studies of their effects and after-effects in laboratory conditions using a wide scale of local aquatic organisms. The selected species of the most sensitive aquatic organisms should include those which are unable to escape the impact of combined action of oil and cleaners. Special attention should be directed to the research of effects of these pollutants on studied organisms at their most sensitive stages of life (reproduction, hatching, early stages of development), as after-effects of exposure to pollutants may be observed in future generations.     

Acute and Chronic Toxicity of Chlorine Dioxide (ClO2) and Chlorite (ClO2ˉ) to Rainbow Trout (Oncorhynchus mykiss) (4 pp)

Goal, Scope and Background Chlorite (ClO2ˉ) is a primary decomposition product when chlorine dioxide (ClO2) is added during water treatment; therefore the toxic effects of both compounds on aquatic organisms are possible. Limited data are available concerning their toxicity to fish. The aim of this study was to investigate sensitivity of rainbow trout to acute and chronic toxicity of chlorine dioxide and chlorite, and to estimate the Maximum-Acceptable-Toxicant-Concentration (MATC) of those compounds in fish. Methods The acute and chronic toxicity of chlorine dioxide and chlorite to larval and adult rainbow trout was investigated in 96-hour to 20-day laboratory exposures evaluating the wide range spectrum of biological indices under semi-static conditions. Results and Discussion Median lethal concentration (96-hour LC50) values derived from the tests were: 2.2 mg/l for larvae; 8.3 mg/l for adult fish and 20-day LC50 for larvae was 1.6 mg/l of chlorine dioxide, respectively. Chlorite was found to be from 48 to 18 times less acutely toxic to larvae and adult fish, correspondingly. Both chemical compounds induced similar toxic effects in rainbow trout larvae during chronic tests (they affected cardio-respiratory and growth parameters), but chlorine dioxide had a higher toxic potency than chlorite. A significant decrease in the heart rate and respiration frequency of larvae was established. However, within an increase in exposure duration recovery of cardio-respiratory responses was seen to have occurred in larvae exposed to chlorite. Meanwhile, in larvae exposed to chlorine dioxide, a significant decrease in cardio-respiratory responses remained during all 20-day chronic bioassays. Chlorine dioxide also more strongly affected growth parameters of rainbow trout larvae at much lower test concentrations. Decreased rate of yolk-sack resorption occurred only in the tests with chlorine dioxide. Conclusions Maximum-Acceptable-Toxicant-Concentration (MATC) of 0.21 mg/l for chlorine dioxide and of 3.3 mg/l for chlorite to fish was derived from chronic tests based on the most sensitive parameter of rainbow trout larvae (growth rate). According to substance toxicity classification accepted for Lithuanian inland waters, chlorine dioxide and chlorite can be referred to substances of \moderate\ toxicity to fish. Recommendations and Outlook Due to its very reactive nature, chlorine dioxide is rapidly (in a few hours) reduced to chlorite, which is persistent also as a biocide but 16 times less toxic to fish, according to MATC. Therefore, it is much more likely that fish will be exposed to chlorite than to chlorine dioxide in natural waters. Presently accepted, the Maximum-Permitted-Concentration of total residual chlorine (TRC) in waste-water discharging into receiving waters is 0.6 mg/l. If this requirement will not be exceeded, it is unlikely that fish would be exposed to lethal or even to sublethal concentrations of chlorine dioxide or chlorite. Furthermore, chlorine dioxide does not generate toxic nitrogenous (chloramines) or carcinogenic organic residuals (trihalomethanes). All these properties make chlorine dioxide a more promising biocide than chlorine.     

Changes in the levels of aspartate and alanine aminotransferase activities of Anabastestudineus (Bloch) exposed to disyston

In $\text{Anabas}$$\text{testudineus}$, the per cent changes noted in the aspartate aminotransferase (AAT) activities of gill, brain, intestine, liver, muscle and kidney until 6 hr of exposure to lethal concentration (10.5 mg/L disyston) were relatively much less when compared to those of sublethal concentration (4.0 mg/L disyston). But in the case of alanine aminotransferase (AIAT) activity, the tissues which showed such effect (paradoxical effect) were gill, brain and liver.     

First Detection of Hepatitis E Virus in Shellfish and in Seawater from Production Areas in Southern Italy

Shellfish samples (n = 384) from production areas, water samples from the same areas (n = 39) and from nearby sewage discharge points (n = 29) were analyzed for hepatitis E virus (HEV) by real-time and nested RT-PCR. Ten shellfish samples (2.6%) and five seawater samples (12.8%) tested positive for HEV; all characterized strains were G3 and showed high degree of sequence identity. An integrated surveillance in seafood and waters is relevant to reduce the risk of shellfish-associated illnesses.