An in vivo evaluation of induction of abnormal sperm morphology in mice by landfill leachates
04 Apr 2005-Mutation Research-genetic Toxicology and Environmental Mutagenesis (Elsevier)-Vol. 582, Iss: 1, pp 28-34
TL;DR: Physico-chemical analysis of the test samples shows that they contained constituents that are capable of inducing mutation in biologic system, which is relevant in environmental waste management, and for the assessment of the hazardous effects of the chemicals in landfill leachates.
Abstract: Although several reports have demonstrated the acutely toxic and genotoxic effects of landfill leachates in microbial organisms, plants and aquatic animals, the effects of pollutants present in these leachates have not been clarified yet in terrestrial animals. This study mainly aimed to evaluate a potential genetic effect of raw and simulated leachates from Orita-Aperin and Oworonsoki landfills in south-west Nigeria by use of the murine sperm-head abnormality test. These landfills neither have a synthetic membrane liner at the bottom, nor a natural layer of compacted soil with the desired hydraulic conductivity, nor a run-off control system. As a result, the leachates produced are discharged into the environment. Samples designated as Orita-Aperin Raw Leachate (OARL), Orita-Aperin Simulated Leachate (OASL), Oworonsoki Raw Leachate (OWRL) and Oworonsoki Simulated Leachate (OWSL) were analyzed in the sperm-head abnormality test at concentrations (v/v) of 1%, 2.5%, 5%, 10% and 25%. Mice were given 0.5 ml sample per day for five consecutive days by intraperitoneal injection. Each dose group comprised seven mice, and a 5-week exposure period was utilized. The data show that the test mixtures induced a dose-dependent, statistically significant increase (P
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TL;DR: An integrated strategy to evaluate the toxicity of the leachate using chemical analyses, risk assessment guidelines and in vitro assays using the hepatoma HepG2 cells as a model was applied on an industrial waste landfill in northern Italy.
Abstract: Solid wastes constitute an important and emerging problem. Landfills are still one of the most common ways to manage waste disposal. The risk assessment of pollutants from landfills is becoming a major environmental issue in Europe, due to the large number of sites and to the importance of groundwater protection. Furthermore, there is lack of knowledge for the environmental, ecotoxicological and toxicological characteristics of most contaminants contained into landfill leacheates. Understanding leachate composition and creating an integrated strategy for risk assessment are currently needed to correctly face the landfill issues and to make projections on the long-term impacts of a landfill, with particular attention to the estimation of possible adverse effects on human health and ecosystem. In the present study, we propose an integrated strategy to evaluate the toxicity of the leachate using chemical analyses, risk assessment guidelines and in vitro assays using the hepatoma HepG2 cells as a model. The approach was applied on a real case study: an industrial waste landfill in northern Italy for which data on the presence of leachate contaminants are available from the last 11 years. Results from our ecological risk models suggest important toxic effects on freshwater fish and small rodents, mainly due to ammonia and inorganic constituents. Our results from in vitro data show an inhibition of cell proliferation by leachate at low doses and cytotoxic effect at high doses after 48 h of exposure.
108 citations
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TL;DR: From the review it appears that there is a need for a multispecies approach to evaluate leachate toxicity, and different bioassays available for assessing the hazard posed by landfill leachates are reviewed.
Abstract: Landfilling is the most common solid waste management practice. However, there exist a potential environmental risk to the surface and ground waters due to the possible leaching of contaminants from the landfill leachates. Current municipal solid waste landfill regulatory approaches consider physicochemical characterization of the leachate and do not assess their potential toxicity. However, assessment of toxic effects of the leachates using rapid, sensitive and cost-effective biological assays is more useful in assessing the risks as they measure the overall toxicity of the chemicals in the leachate. Nevertheless, more research is needed to develop an appropriate matrix of bioassays based on their sensitivity to various toxicants in order to evaluate leachate toxicity. There is a need for a multispecies approach using organisms representing different trophic levels so as to understand the potential impacts of leachate on different trophic organisms. The article reviews different bioassays available for assessing the hazard posed by landfill leachates. From the review it appears that there is a need for a multispecies approach to evaluate leachate toxicity.
101 citations
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TL;DR: In this paper, the authors focus on leachate composition, plume migration, and contaminant façade contamination in landfill leachates and its treatment prior to disposal into the environment.
Abstract: Landfills are the primary option for waste disposal all over the world. Most of the landfill sites across the world are old and are not engineered to prevent contamination of the underlying soil and groundwater by the toxic leachate. The pollutants from landfill leachate have accumulative and detrimental effect on the ecology and food chains leading to carcinogenic effects, acute toxicity, and genotoxicity among human beings. Management of this highly toxic leachate presents a challenging problem to the regulatory authorities who have set specific regulations regarding maximum limits of contaminants in treated leachate prior to disposal into the environment to ensure minimal environmental impact. There are different stages of leachate management such as monitoring of its formation and flow into the environment, identification of hazards associated with it, and its treatment prior to disposal into the environment. This article focuses on: (i) leachate composition, (ii) plume migration, (iii) contaminant fa...
98 citations
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TL;DR: Wood mice showed lower bioaccumulation of metals and lower variation caused by biotic factors, and the morphological and physiological alterations demonstrated that they were also more sensitive at environmental pollution, which indicates that the landfill affects the health of wild mice.
Abstract: We assess the bioaccumulation of metals (Pb, Hg, Cd, Fe, Mg, Zn, Cu, Mn, Mo, Cr) and effects of landfill leachates on morphological (RI, relative weights), plasma (GPT, GOT, creatinine), and genotoxic (MNT) parameters in wood mice, Apodemus sylvaticus, inhabiting close the Garraf landfill site (NE Spain). Due to the high age- and sex-dependent variation in wild populations, we also studied the effect of these biotic factors on the parameters studied. Wood mice from the landfill site, sited in a partially protected area, showed more Cd, Fe, Zn, Cu, Mn, Mo, and Cr than specimens from the reference site. Moreover, mice near the landfill registered low RI and high relative renal weight, GPT, and MN frequency, which indicate that the landfill affects the health of wild mice. In contrast to sympatric shrews from a previous study, wood mice showed lower bioaccumulation of metals and lower variation caused by biotic factors. Moreover, the morphological and physiological alterations demonstrated that they were also more sensitive at environmental pollution. Given the contribution of small mammals to ecosystem function and the scarce ecotoxicological data on the effects of landfill pollution on wild terrestrial mammals, we consider that our study can be used to improve the management of this protected area.
86 citations
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TL;DR: The statistical analysis and spatial variations indicated the leaching of metals from the landfill to the groundwater aquifer system, a potential hazard to both surrounding ecosystems and human populations.
Abstract: Release and transport of leachate from municipal solid waste landfills pose a potential hazard to both surrounding ecosystems and human populations. In the present study, soil, groundwater, and surface water samples were collected from the periphery of a municipal solid waste landfill (located at Ranital of Jabalpur, Madhya Pradesh, India) for laboratory analysis to understand the release of contaminants. The landfill does not receive any solid wastes for dumping now as the same is under a landfill closure plan. Groundwater and soil samples were collected from the bore holes of 15m deep drilled along the periphery of the landfill and the surface water samples were collected from the existing surface water courses near the landfill. The landfill had neither any bottom liner nor any leachate collection and treatment system. Thus the leachate generated from the landfills finds paths into the groundwater and surrounding surface water courses. Concentrations of various physico-chemical parameters including some toxic metals (in collected groundwater, soil, and surface water samples) and microbiological parameters (in surface water samples) were determined. The analyzed data were integrated into ArcGIS environment and the spatial distribution of the metals and other physic- chemical parameter across the landfill was extrapolated to observe the distribution. The statistical analysis and spatial variations indicated the leaching of metals from the landfill to the groundwater aquifer system. The study will help the readers and the municipal engineers to understand the release of contaminants from landfills for better management of municipal solid wastes.
83 citations
References
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TL;DR: The results suggest that sperm abnormalities might provide a rapid inexpensive mammalian screen for agents that lead to errors in the differentiation of spermatogenic stem cells in vivo and thus indicate agents which might prove to be mutagenic, teratogenic, or carcinogenic.
Abstract: The sperm of (C57BL X C3H)F1 mice were examined 1, 4, and 10 weeks after a subacute treatment with one of 25 chemicals at two or more dose levels. The fraction of sperm that were abnormal in shape was elevated above control values of 1.2-3.4% for methyl methanesulfonate, ethyl methanesulfonate, griseofulvin, benzo[a]pyrene, METEPA [tris(2-methyl-l-aziridinyl)phosphine oxide], THIO-TEPA [tris(l-aziridinyl)phosphine sulfide], mitomycin C, myleran, vinblastine sulphate, hydroxyurea, 3-methylcholanthrene, colchicine, actinomycin D, imuran, cyclophosphamide, 5-iododeoxyuridine, dichlorvos, aminopterin, and trimethylphosphate. Dimethylnitrosamine, urethane, DDT [1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane], 1,1-dimethylhydrazine, caffeine, and calcium cyclamate did not induce elevated levels of sperm abnormalities. The results suggest that sperm abnormalities might provide a rapid inexpensive mammalian screen for agents that lead to errors in the differentiation of spermatogenic stem cells in vivo and thus indicate agents which might prove to be mutagenic, teratogenic, or carcinogenic.
811 citations
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TL;DR: The mouse sperm morphology test has potential use for identifying chemicals that induce spermatogenic dysfunction and perhaps heritable mutations, and is found to be highly sensitive to germ-cell mutagens.
Abstract: The literature on the mouse sperm morphology test and on other sperm tests in nonhuman mammals was reviewed (a) to evaluate the relationship of these tests to chemically induced spermatogenic dysfunction, germ-cell mutagenicity, and carcinogenicity, and (b) to make an interspecies comparison to chemicals. A total of 71 papers were reviewed. The mouse sperm morphology test was used to assess the effects of 154 of the 182 chemical agents covered. 4 other murine sperm tests were also used: the induction of acrosomal abnormalities (4 agents), reduction in sperm counts, (6 agents), motility (5 agents), and F1 sperm morphology (7 agents)). In addition, sperm tests for the spermatogenic effects of 35 agents were done in 9 nonmurine mammalian species; these included analyses for sperm count, motility, and morphology, using a large variety of study designs. For the mouse sperm morphology test, 41 agents were judged by the reviewing committee to be positive inducers of sperm-head shape abnormalities, 103 were negative, and 10 were inconclusive. To evaluate the relationship between changes in sperm morphology and germ cell mutagenicity, the effects of 41 agents on mouse sperm shape were compared to available data from 3 different mammalian germ-cell mutational tests (specific locus, heritable translocation, and dominant lethal). The mouse sperm morphology test was found to be highly sensitive to germ-cell mutagens; 100% of the known mutagens were correctly identified as positives in the sperm morphology test. Data are insufficient at present to access the rate of false positives. Although it is biologically unclear why one might expect changes in sperm morphology to be related to carcinogenesis, we found that (a) a positive response in the mouse sperm morphology test is highly specific for carcinogenic potential (100% for the agents surveyed), and (b) overall, only 50% of carcinogens were positive in the test (i.e., sensitivity approximately equal to 50%). Since many carcinogens do not produce abnormally shaped sperm even at lethal doses, negative findings with the sperm test cannot be used to classify agents as noncarcinogens. We conclude that the mouse sperm morphology test has potential use for identifying chemicals that induce spermatogenic dysfunction and perhaps heritable mutations. Insufficient numbers of chemicals agents have been studied by the other sperm tests to permit similar comparisons. A comparison of 25 chemicals tested with sperm counts, motility, and morphology in at least 2 species (including man, mouse and 9 other mammals) demonstrated good agreement in response among species. With further study, interspecies comparisons of chemically induced sperm changes may be useful for predicting and evaluating human effects.
380 citations
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TL;DR: The premeiotic synthesis of DNA takes place in primary spermatocytes during the resting phase and terminates just before the visible onset of the meiotic prophase.
Abstract: Mice were injected intraperitoneally with 15 µc of H3-thymidine. The time course of the labeling in spermatogonia and spermatocytes was studied by using autoradiography on 5 µ sections stained by the periodic acid-Schiff method and hematoxylin over a period of 57 hours after injection. Four generations of type A (called AI, AII, AIII, and AIV), one of intermediate, and one of type B spermatogonia occur in one cycle of the seminiferous epithelium. The average life span is about the same in all spermatogonia, i.e., about 27 to 30.5 hours. The average pre-DNA synthetic time, including the mitotic stages from metaphase through telophase and the portion of interphase preceding DNA synthesis, is also not very different, ranging between 7.5 and 10.5 hours. A remarkable difference exists, however, in the duration of DNA synthesis and of the post-DNA synthetic period. The average DNA synthetic time is very long and is highly variable in type B (14.5 hours), a little shorter and less variable in intermediate (12.5 hours) and AIV (13 hours) spermatogonia, and much shorter and very constant in AIII (8 hours), AII and AI (7 to 7.5 hours) spermatogonia. Conversely, the average post-DNA synthetic time, corresponding essentially to the duration of the prophase, is short and very constant in type B (4.5 hours), longer and variable in intermediate (6 hours) and AIV (8 hours) spermatogonia, and much longer and much more variable in AIII (11 hours), AII and AI (14 hours) spermatogonia. The premeiotic synthesis of DNA takes place in primary spermatocytes during the resting phase and terminates just before the visible onset of the meiotic prophase. Its average duration is 14 hours. No further synthesis of DNA takes place in later stages of spermatogenesis.
327 citations
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TL;DR: The evaluation of hazardous wastes and effluents by genotoxicity assays may provide data useful not only for hazard identification but for comparative risk assessment.
Abstract: A review of the literature published on the genotoxicity of industrial wastes and effluents using short-term genetic bioassays is presented in this document. The importance of this task arises from the ubiquity of genotoxic compounds in the environment and the need to identify the sources of contamination so that efforts aimed at control and minimization can be implemented. Of even greater significance is the immediate concern for the welfare of human health and the environment. Subheadings of this document include a description of the genetic bioassays that have been used to test industrial wastes, a compendium of methods commonly used to prepare crude waste samples for bioassay, and a review of the genetic toxicity of wastes and effluents. Wastes and effluents have been grouped according to industrial source. Major categories include chemical and allied products, pulp and paper manufacturing, defense and munitions, petroleum refining, primary metal industries, and miscellaneous industrial manufacturers. Within each industrial category, a synopsis of individual genetic toxicity studies is presented, followed by an interpretation of results on a comprehensive, industry-wide basis. In this evaluation, a discussion of the types and extent of genotoxic damage caused by a particular set of wastes is presented, and potential sources of genotoxic activity are identified. Concluding the document is a commentary, which discloses potential shortcomings in the way in which current legislation protects human heath and the environment from the release of genotoxic substances via industrial wastes and effluents. It also provides an assessment of the genotoxic burden that industrial wastes place on the environment.
302 citations
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TL;DR: Study of the association of the somatic and gametic genotypes with the gametic phenotype has been termed the genetics of gametes.
Abstract: Summary
1. Study of the association of the somatic and gametic genotypes with the gametic phenotype has been termed the genetics of gametes. The latter is closely associated with problems of fertility and infertility; with the general validity of the postulate of random union of the gametes; with experimental attempts to control sex ratio or other Mendelian segregations; and with the study of chromosomal anomalies. Effects of the post-segregational genotype (after meiosis) are of exceptional interest.
2. The dimensions of mammalian spermatozoa show numerous patterns of genetic behaviour, such as strain and breed differences, additive variation, heterosis, high heritability, maternal effects, and a tentative ‘paternal cytoplasmic effect’. The high degree of additive genetic determination is reflected in the smooth progress of a genetic selection programme that brought into existence strains of mice with either long or short spermatozoan midpieces. The balance sheet of variation in mammalian spermatozoan dimensions is as follows. Apart from the natural variation between individual spermatozoa of a male, the paramount factors are the genetic ones. Variation is extraordinarily independent of other biological sources of variation and of environmental ones. The high heritability of most spermatozoan dimensions suggests that they are not closely associated with reproductive fitness, whereas a measure of fitness (resistance of spermatozoa to eosin staining) shows a low heritability.
3. Post-segregational gene action has been sought but not conclusively demonstrated in attempts to recognize X- and Y-bearing spermatozoa visually, and in attempts to control sex ratio by separating them according to electrophoretic or sedimentation behaviour.
4. Variations in the DNA content of spermatozoa, and gross morphological defects, are associated with infertility and their incidence is often genetically controlled.
5. There is controversy over the balance between specific spermatozoan antigens and ‘coating antigens’ acquired from the seminal fluid. Reports that AB individuals (heterozygous for blood-group antigens) produce phenotypically A and B spermatozoa would indicate post-segregational gene action but are also controversial. Sex ratio has not yet been controlled by subjecting spermatozoa to anti-Y-chromosome antibodies. Genetic (strain) differences exist in spermatozoan antigens.
6. The incidence of non-eosinophil spermatozoa is a measure of semen fertility. Strain and breed differences exist but the heritability is low. An absence of demonstrable genetic variation in spermatozoan motility may be due to a swamping effect of non-genetic factors.
7. Diploid spermatozoa differ genetically and phenotypically from the normal haploid ones. The incidence of diploid spermatozoa is associated with infertility and is controlled genetically. The incidence of polyspermy and of supplementary spermatozoa are also controlled genetically.
8. Heterospermic (mixed) insemination from two or more sires discriminates efficiently between the sires in terms of the relative numbers of offspring produced. One experiment showed a genetic control (strain differences) in heterospermic performance. Heterospermic performance of a sire is constant over periods of time and is a predictor of the relative fertility obtained in ‘ordinary’ fertility tests. Heterospermic insemination per se has no apparent practical value.
9. Species crosses in animals tend to produce diploid oocytes and triploid embryos. ‘Foreign’ mammalian eggs or spermatozoa did not fertilize in the same proportion as native ones.
10. The most recent determination shows a I : I sex ratio in the very early embryo.
11. Mammals are scarcely likely to be exempt from certain unorthodox genetic mechanisms, but positive evidence is only indicative. Micro-organisms have been detected and sometimes have a genetic role in spermatozoa and eggs of animals. There is a theory that mitochondria (and therefore most of the spermatozoan midpiece) are modified bacteria. A tentative ‘paternal cytoplasmic effect’ on mouse spermatozoan dimensions, mediated by mitochondria1 inheritance, has been invoked. Conditions may exist for possible transfer of genetic information from spermatozoa to body cells via leucocytes. There is no confirmation of experiments for transfer of genetic information by injection of DNA of specific genotypes prepared from spermatozoa or other tissues.
12. The behaviour of ‘tailless’ alleles in mice provides a unique example of post-segregational gene action, the t-spermatozoa of heterozygous Tt males yielding more offspring than the T-spermatozoa. This work explains the aberrant transmission of t through the male parent, and also the high frequency of the allele in wild populations. It is the best example of post-segregational gene action in animals, provides an exception to the random union of gametes, and permits a novel way of controlling the transmission of genetic information between generations. There is presumptive post-segregational gene action in other segregations in mouse and man.
13. The study of heteroploidy (abnormal chromosome numbers) provides massive evidence of anomalies in the mammalian egg that are in themselves genetic (e.g. diploidy) and phenotypic (e.g. abnormal numbers of polar bodies or pronuclei); that come into being under genetic control, and that have genetic consequences (e.g. triploidy) and phenotypic ones (e.g. inviability) for the embryo. This is illustrated by suppression of the cytoplasmic second meiotic division of the egg, giving diploid parthenogenetic embryos from unfertilized eggs and triploid ones from fertilized eggs. Triploidy is the dominant form of polyploidy in vertebrates. Triploidy is lethal in the mammalian embryo and is a main cause of the ‘natural’ level of prenatal mortality in man.
14. The groups of cells produced by meiosis in both sexes of animals are virtually syncytial. This has a bearing on post-segregational heterosis and gene effects.
15. The unique and apparently indisputable instance of post-segregational gene action in ‘tailless’ mice is to be contrasted on the one hand with numerous persuasive arguments (especially effective in Drosophila) that such action cannot or does not occur in animals or else is mimicked by another phenomenon, meiotic drive; and on the other hand contrasts with an hypothesis that post-segregational gene action occurs on a gigantic scale in all organisms in which crossing-over takes place. It is concluded for the time being that post-segregational gene action does occur in mammals, but is rare. Because of the rarity, a reasonable first interpretation in all gamete genetics is that gene action is mediated by the somatic genotype, unless there are specific reasons to the contrary. There is urgent need for a technique whereby the biological potential of a given spermatozoon can be recognized by fertilizing a given egg with it.
16. As a series of working hypotheses, it is proposed that the general architecture and course of differentiation of gametes are determined by the genotype of the soma and the same pre-segregational genotype of the germ line, the initial gene action being a synthesis of messenger RNA before meiosis in the germ line or at any time in the soma. These effects of the somatic genotype bring the gamete into existence and are also responsible for most of the widespread variation in its phenotype. The gamete so produced can be varied by a very few genes or sets of genes synthesizing messenger RNA in the germ cells after segregation, or else exerting action in some other way connected with their direct physical and chemical nature or their arrangement. These post segregational effects are more likely to be mediated by whole blocs of genes, especially heterochromatic and nucleolar ones, than by ‘billiard-ball’ single genes in the narrow sense.
175 citations