Ecotoxicological evaluation of municipal sludge

Municipal wastes originating from urban and industrial areas have become a major source of soil, ground and surface water pollution. These undesirable agents in our environment significantly interact with our flora and fauna. The aim of this study was to test samples of municipal sludge (MS) for their ecotoxicological potential by using sensitive bioassays involving a plant, Vicia faba, and the earthworm, Eisenia foetida. A 10% leachate of MS was prepared for the experiments, and V. faba seedlings were exposed to three leachate concentrations (2.5%, 5% and 10%) for 5 days. The findings revealed chromosome aberrations during the metaphase as well as the anaphase of cell division, and inhibition of the mitotic index, which reflects that MS originating from domestic and other human activities may be genotoxic to the living organisms of the ecosystem. Abnormalities in chlorophyll content, plant growth, root length, shoot length and root/shoot length ratio in V. faba clearly indicated the toxicity of the sludge. Behavioural and reproduction studies with E. foetida also provided evidence for the toxic nature of the MS.     

Municipal sludge leachate-induced genotoxicity in mice--a subacute study

Inappropriate disposal of municipal sludge (MS) results in the leaching of toxic metals and organic chemicals, which can contaminate the surface and ground water leading to the serious health hazards. In this study, the genotoxic potential of the leachate prepared from MS sample was examined in mouse bone marrow cells through chromosomal aberrations (CA), micronucleus test (MT) and comet assay. Analysis of metals and physicochemical parameters of the leachate was also carried out to correlate the genotoxic results. The dried sludge showed high concentrations of heavy metals, viz. Cr, Cu, Pb and Ni. However, in 10% leachate, concentrations of these metals were manifold lower than that of obtained in dried sludge. Male mice orally gavaged to leachates (0.1-0.4 ml/mouse/day) for 15 days revealed significant (P<0.01, P<0.001) inhibition of mitotic index (MI) and induction of chromatid/chromosome fragments and breaks in all the treatment groups. The effect was observed to be dose-dependent. Treatment of mice with leachates also induced significant (P<0.001) frequencies of micronucleated polychromatic erythrocytes (MNPCE). The results of comet assay revealed a statistically significant (P<0.05 and <0.01) DNA damage in bone marrow cells exposed to 0.2-0.4 ml/mouse/day. Findings of the present study indicate that the constant exposure of MS leachate can cause genotoxic effects in mammals and suggest risks in human population.     

Municipal sludge-induced phytotoxicity

Municipal sludge (MS), containing various types of environmental pollutants, can exert phytotoxic effects in plants. Seed germination, chlorophyll content, plant weight, root and shoot growth, and metal levels, were measured as endpoints to assess the phytotoxic effects of leachates of MS from Lucknow city in three plants, Triticum aestivum, Brassica campestris and Phaseolus aureus. Among the five metals analysed, lead was found to be present at the highest concentration, and chromium at the lowest. Aqueous MS leachates had lower amounts of metals than pure MS. Seed germination was inhibited significantly in all three plants. The greatest inhibition of root and shoot growth was observed in B. campestris and T. aestivum. Total chlorophyll was depleted in all three plants, but the maximum depletion was observed in T.aestivum exposed to 10% leachate. Symptoms such as necrosis, leaf discoloration and root curling were also observed after exposure to 10% leachate, and plant weight was reduced significantly. It is evident that MS from Lucknow city exerted phytotoxic effects on all three test plant species.     

Cytogenetic and microtubule array effects of the zineb-containing commercial fungicide formulation Azzurro(®) on meristematic root cells of Allium cepa L

Zineb [ethylene bis(dithiocarbamate) zinc] is a widely employed foliar fungicide for agricultural and industrial applications. Allium cepa L. is a reliable model for the assessment of xenobiotic genotoxicity and cytotoxicity. We evaluated the effects of the zineb-containing commercial formulation Azzurro(?) (70% zineb) in cell cycle stages of the meristem root cells of A. cepa. The mitotic index (MI), chromosomal aberrations at anaphase/telophase (CAs), micronuclei (MN), and abnormalities in immunodetected microtubule structures, e.g., preprophasic band (PPB), mitotic spindle (MS), and phragmoplast (Phrag), were used as end-points. Azzurro(?) (1 and 10μg/ml) induced a significant increase in the frequency of CAs (P<0.05), and the higher concentration inhibited the MI (P<0.05) compared to control values. The frequency of MN did not differ from control values at any concentration. Treatment with 1μg/ml Azzurro(?) induced a significant increase in the frequency of abnormal PPB (P<0.01), MS (P<0.001), and Phrag (P<0.01) and, at 10μg/ml, enhancements in the frequencies of abnormal MS (P<0.05) and Phrag (P<0.05) were seen. A tubulin immunodetection assay showed that exposure to Azzurro(?) interferes with normal assembly of microtubule structures during mitosis.     

Co-digestion of intermediate landfill leachate and sewage sludge as a method of leachate utilization

This study examines the co-digestion of intermediate landfill leachate and sewage sludge from a municipal wastewater treatment plant. Application of leachate as a co-fermentation component increased the concentrations of soluble organic compounds (expressed as total organic carbon), ammonium nitrogen, and alkalinity in the digester influents.The biogas yield obtained from the co-fermentation of a 20:1 sewage sludge: intermediate leachate mixture was 1.30 m³ per kg of removed volatile solids (VS), while that from a 10:1 mixture was 1.24 m³ per kg of removed VS. These values exceeded the biogas yield for the sludge alone by 13% and 8%, respectively. The leachate addition influenced the proportion of methane to a minor extent. Increased methane yields of 16.9% and 6.2% per kg of removed VS were found for the two sewage sluge:intermediate leachate mixtures, respectively.     

Genotoxicity of five food preservatives tested on root tips of Allium cepa L

The effects of the food preservatives sodium benzoate (SB), boric acid (BA), citric acid (CA), potassium citrate (PC) and sodium citrate (SC) have been studied on root tips of Allium cepa L. Roots of A. cepa were treated with a series of concentrations, ranging from 20 to 100 ppm for 5, 10 and 20 h. The results indicate that these food preservatives reduced mitotic division in A. cepa compared with the respective control. Mitotic index values were generally decreased with increasing concentrations and longer treatment times. Additionally, variations in the percentage of mitotic stages were observed. The total percentage of aberrations generally increased with increasing concentrations of these chemicals and the longer period of treatment. Different abnormal mitotic figures were observed in all mitotic phases. Among these abnormalities were anaphase bridges, C-mitosis, micronuclei, lagging, stickiness, breaks and unequal distribution.     

Toxicological evaluation of three contaminants of emerging concern by use of the Allium cepa test

Di(2-ethylhexyl)phthalate, triclosan and propylparaben are contaminants of emerging concern that have been subjected to extensive toxicological studies, but for which limited information is currently available concerning adverse effects on terrestrial plant systems. The Allium cepa test, which is considered one of the most efficient approaches to assess toxic effects of environmental chemicals, was selected to evaluate the potential risks of these ubiquitous pollutants. Our data demonstrate that all three compounds studied may in some way be considered toxic, but different effects were noted depending on the chemical and the end point analysed. Results derived from the analysis of macroscopic parameters used in testing for general toxicity, revealed that while di(2-ethylhexyl)phthalate had no apparent effects, the other two chemicals inhibited A. cepa root growth in a dose-dependent manner. On the other hand, although all three compounds caused alterations in the mitotic index of root-tip cells, propylparaben was the only one that did not show evidence of genotoxicity in assays for chromosome aberrations and micronuclei. The results of the present study clearly indicate that sensitive plant bioassays are useful and complementary tools to determine environmental impact of contaminants of emerging concern.     

Cytotoxic and genotoxic effects of Br-containing oxaphosphole on Allium cepa L. root tip cells and mouse bone marrow cells

The continuous production and release of chemicals into the environment has led to the need to assess their genotoxicity. Numerous organophosphorus compounds with different structures have been synthesized in recent years, and several oxaphosphole derivatives are known to possess biological activity. Such chemical compounds may influence proliferating cells and cause disturbances of the genetic material. In this study, we examined the cytotoxicity and genotoxicity of 4-bromo-N,N-diethyl-5,5-dimethyl-2,5-dihydro-1,2-oxaphosphol-2-amine 2-oxide (Br-oxph). In A. cepa cells, Br-oxph (10(-9) M, 10 (-6) M and 10 (-3) M) reduced the mitotic index 48 h after treatment with the two highest concentrations, with no significant effect at earlier intervals. Mitotic cells showed abnormalities 24 h and 48 h after treatment with the two lowest concentrations but there were no consistent changes in interphase cells. Bone marrow cells from mice treated with Br-oxph (2.82 x 10 (-3) μg/kg) also showed a reduced mitotic index after 48 h and a greater percentage of cells with aberrations (principally chromatid and isochromatid breaks). These findings indicate the cytotoxicity and genotoxicity of Br-oxph in the two systems studied.     

Enhancement of hydrolysis and acidification of solid organic waste by a rotational drum fermentation system with methanogenic leachate recirculation

A cascade process of a rotational drum fermentation system with leachate recirculation from a methanogenic to the acidogenic reactor was constructed to enhance the hydrolysis and acidification of solid organic waste. Using fresh soybean meal as substrates, two process configurations, Cascade process 1 and 2, without and with leachate recirculation, respectively were employed to perform the experimental estimation under mesophilic condition and a total HRT of 20 days. An apparent first-order hydrolysis rate constant of 9.0 x 10(-3)/d for Cascade process 1 at pH 4.5-4.6, and 15.8 x 10(-3)/d for Cascade process 2 at pH 4.6-5.2 were obtained. The apparent VS degradation ratios ranged from 16.5% to 21.1% and total VA (as acetic acid) from 14.5 to 16.7 g/L. Occupying ratios for ionized VA decreased from 40.5% to 35.3% for Cascade process 1 and increased to 68.5% for Cascade process 2. However, occupying ratios of acetic acid decreased from 96.1% to 94.3% for Cascade process 1 and to 72.6% for Cascade process 2, whereas propionic acid and butyric acid ratios increased in acidogenesis of Cascade process 2. The leachate recirculation promoted hydrolysis of substrate in Cascade process 2, where apparent hydrolysis rate constant and VS degradation ratio were higher than that of Cascade process 1.     

Genotoxicity assessment of a pharmaceutical effluent using four bioassays

Pharmaceutical industries are among the major contributors to industrial waste. Their effluents when wrongly handled and disposed of endanger both human and environmental health. In this study, we investigated the potential genotoxicity of a pharmaceutical effluent, by using the Allium cepa, mouse- sperm morphology, bone marrow chromosome aberration (CA) and micronucleus (MN) assays. Some of the physico-chemical properties of the effluent were also determined. The A. cepa and the animal assays were respectively carried out at concentrations of 0.5, 1, 2.5, 5 and 10%; and 1, 5, 10, 25 and 50% of the effluent. There was a statistically different (p < 0.05), concentration-dependent inhibition of onion root growth and mitotic index, and induction of chromosomal aberrations in the onion and mouse CA test. Assessment of sperm shape showed that the fraction of the sperm that was abnormal in shape was significantly (p < 0.05) greater than the negative control value. MN analysis showed a dose-dependent induction of micronucleated polychromatic erythrocytes across the treatment groups. These observations were provoked by the toxic and genotoxic constituents present in test samples. The tested pharmaceutical effluent is a potentially genotoxic agent and germ cell mutagen, and may induce adverse health effects in exposed individuals.     

Genotoxicity of drinking water disinfection by-products (bromoform and chloroform) by using both Allium anaphase-telophase and comet tests

Genotoxic effects of bromoform and chloroform, disinfection by-products of the chlorination of drinking water, were examined by using mitotic index (MI), mitotic phase, chromosome aberrations (CAs) and comet assay on root meristematic cells of Allium cepa. Different concentrations of bromoform (25, 50, 75 and 100 μg/mL) and chloroform (25, 50, 100 and 200 μg/mL) were introduced to onion tuber roots. Distilled water was used as a negative control and methyl methansulfonate (MMS-10 μg/mL) as positive control. All obtained data were subjected to statistical analyses by using SPSS 15.0 for Windows software. For comparison purposes, Duncan multiple range tests by using one-way analysis of variance were employed and p < 0.05 was accepted as significant value. Exposure of both chemicals (except 25 μg/mL applications of bromoform) significantly decreased MI. Bromoform and chloroform (except 25 μg/mL applications) increased total CAs in Allium anaphase-telophase test. A significant increase in DNA damage was also observed at all concentrations of both bromoform and chloroform examined by comet assay. The damages were higher than that of positive control especially at 75–100 μg/mL for bromoform and 100–200 μg/mL for chloroform.     

Clastogenicity of pentachlorophenol, 2,4-D and butachlor evaluated by Allium root tip test

The meristematic mitotic cells of Allium cepa is an efficient cytogenetic material for chromosome aberration assay on environmental pollutants. For assessing genotoxicity of pentachlorophenol (PCP), 2,4-dichlorophenoxyacetic acid (2,4-D) and 2-chloro-2,6-diethyl-N-(butoxymethyl) acetanilide (butachlor), 50% effective concentration (EC(50)), c-mitosis, stickiness, chromosome breaks and mitotic index (MI) were used as endpoints of genotoxicity. EC(50) values for PCP and butachlor are 0.73 and 5.13 ppm, respectively. 2,4-D evidently induced morphological changes at higher concentrations. Some changes like crochet hooks, c-tumours and broken roots were unique to 2,4-D at 5-20 ppm. No such abnormalities were found in PCP and butachlor treated groups, however, root deteriorated and degenerated at higher concentrations (<3 ppm) in PCP. MI in 2,4-D showed a low average of 14.32% followed by PCP (19.53%), while in butachlor it was recorded 71.6%, which is near to the control value. All chemicals induced chromosome aberrations at statistically significant level. The highest chromosome aberration frequency (11.90%) was recorded in PCP at 3 ppm. Large number of c-mitotic anaphases indicated that butachlor acts as potent spindle inhibitor, whereas, breaks, bridges, stickiness and laggards were most frequently found in PCP showing that it is a potent clastogen.     

pH值和产甲烷抑制剂对两相耦合系统污泥发酵定向产乙酸的影响

采用产氢产乙酸/同型产乙酸两相耦合工艺对剩余污泥进行了半连续式厌氧发酵,主要研究了pH值和产甲烷抑制剂2-bromoethanesulphonate(BES)对耦合系统定向产乙酸的影响.结果表明:碱性pH(pH=10.0)和添加BES都能促进A相乙酸的积累,提高乙酸的产率,同时碱性pH比添加BES更有利于污泥的水解.当...     

添加污泥对尾矿砂理化性质及香樟生理特性的影响

以香樟作为指示植物,选取黄岩、临海和路桥地区污水处理厂污泥,将污泥与尾矿砂按(污泥质量比例为0%对照、25%、50%和75%)配比进行栽培试验。测定添加污泥对尾矿砂理化性质以及香樟生理特性的影响。结果表明:随着污泥比例的增加,混合基质中的有机质、全氮、全磷明显增加,pH值明显降低,离子交换量明显减少,Cu和Cd总量明显增加,而Pb总量明显减少,Zn总量没有明显变化,Cu、Cd和Zn的DTPA提取量明显增加,而Pb的DTPA提取量明显减少。黄岩和临海污泥在25%和50%比例时,香樟叶和茎的生物量和叶绿素含量明显增加,而根的生物量没有明显变化,在75%比例时,生物量和叶绿素含量均明显减少;而添加路桥污泥使香樟叶、茎和根的生物量和叶绿素含量明显减少。丙二醛含量则与生物量和叶绿素含量呈现相反的变化特征。黄岩和临海污泥在25%和50%比例时,根和叶的Cu、Cd、Pb和Zn含量明显减少,在75%比例时,Cu、Cd和Zn含量则明显增加;而添加路桥污泥使叶和根的Cu、Cd和Zn含量明显增加,Pb含量明显减少。研究表明添加污泥提高了尾矿砂的养分含量,同时改变了其重金属组成,对污泥重金属含量和有机质组成的监控可以准确地预测污泥改良后尾矿砂对植物毒性的变化。     

The effect of lead on Allium cepa L.

The effect of lead on Allium cepa L. at concentrations of 0.1, 1.0, 10, 50, 100 and 200 ppm were studied. Analysis focused on root growth, frequency of mitosis in a meristematic zone, and chromosomal aberrations. It was observed that lead reduces root growth and the frequency of mitotic cells in meristematic zones, and increases the frequency of aberrant cells. The intensity of the effects is a function of lead concentration.     

Anaerobic co-digestion of algal sludge and waste paper to produce methane

The unbalanced nutrients of algal sludge (low C/N ratio) were regarded as an important limitation factor to anaerobic digestion process. Adding high carbon content of waste paper in algal sludge feedstock to have a balanced C/N ratio was undertaken in this study. The results showed adding 50% (based on volatile solid) of waste paper in algal sludge feedstock increased the methane production rate to 1170+/-75 ml/l day, as compared to 573+/-28 ml/l day of algal sludge digestion alone, both operated at 4 g VS/l day, 35 degrees C and 10 days HRT. The maximum methane production rate of 1607+/-17 ml/l day was observed at a combined 5 g VS/l day loading rate with 60% (VS based) of paper adding in algal sludge feedstock. Results suggested an optimum C/N ratio for co-digestion of algal sludge and waste paper was in the range of 20-25/1.     

Cycloheximide genotoxicity in in vitro and in vivo test systems

The aim of this study was to investigate if there was any genotoxic effect produced by the antibiotic cycloheximide, widely used as a fungicide in agriculture as well as in everyday laboratory practice. The battery of test systems included the bacterium Salmonella typhimurium (strains TA98 and TA100), the yeast Saccharomyces cerevisiae (D7), Allium cepa somatic cells and mouse bone marrow cells. This combination of test systems enabled us to establish possible effects caused by cycloheximide at different levels of the genome and to indicate a possible mechanism of action. The results obtained in experiments showed that cycloheximide did not induce frameshift or base-pair substitution mutations in S. typhimurium regardless of metabolic activation. In S. cerevisiae cycloheximide had only toxic effects but no increase of mitotic gene conversion was noticed under the conditions of the experiment. However, in A. cepa somatic cells as well as in mouse bone marrow cells cycloheximide showed its activity causing different genetic damages, e.g., chromosome breaks, mitotic disturbances and nuclear abnormalities.     

Toxicity and genotoxicity of wastewater from gasoline stations

The toxicity and genotoxicity of wastewater from eight gasoline stations in Brasília, Brazil's capital city, was studied by assessing chromosomal aberrations, chromosomal malsegregation and the mitotic index in Alliumcepa root cells, and the occurrence of micronucleus and nuclear abnormalities in peripheral erythrocytes of tilapia fish (Oreochromis niloticus). The content of gasoline station effluents was also analyzed based on several physico-chemical parameters. None of the wastewater samples was genotoxic to A. cepa root cells, although cell proliferation was significantly inhibited, especially at the highest concentrations. Likewise, no micronuclei were observed in O. niloticus peripheral erythrocytes, even after exposure to high concentrations, but there was an increase in the number of nuclear abnormalities and fish mortality. These results show that although the effluent from gasoline stations is processed by an oil/water separation system before being discharged into the main sewage system, the wastewater still contains toxic compounds.     

Inhibitory effect of adrenaline and hydrocortisone on the growth of Allium cepa roots

The roots of Allium cepa were allowed to grow in distilled water containing 10(-4) M adrenaline hydrochloride or 2 X 10(-3) M hydrocortisone sodium succinate. Adrenaline inhibited the growth of roots; they decreased in length, number and total dry weight. The total amount of DNA in the roots was reduced much less than that of extracellular root components after adrenaline. Also hydrocortisone treatment resulted in a considerable decrease of the length and dry weight of Allium cepa roots. Both DNA and extracellular root components were influenced.     

Influence of inoculum activity on the bio-methanization of a kitchen waste under different waste/inoculum ratios

The use of a granular inoculum prevented acidification during the anaerobic batch biodegradation of a kitchen waste for waste/inoculum ratios in the range of 0.5–2.3 g VS/g VS, when the alkalinity/COD ratio was 37 mg NaHCO₃/g COD. In similar experiments but using a suspended sludge with a significantly lower activity, the methane production rates and the biodegradability were significantly lower and the pH decreased below 5.5 at the waste/inoculum ratio of 2.3 g VS/g VS. When the added alkalinity was decreased to 2 mg NaHCO₃/g COD, the ratio waste/inoculum was clearly more important than the inoculum activity, since, irrespective of the sludge used, acidification occurred at waste/inoculum ratios higher than 0.5 g VS/g VS. The advantage of using granular sludge was further investigated in order to define reasonable condition of waste/inoculum ratio and added alkalinity that could be applied in practice. For a waste/inoculum ratio of 1.35, there were no significant differences between the results obtained for the biodegradability and maximum methane production rate (MMPR), when the alkalinity decreased from 44 to 22 mg NaHCO₃/g COD.