EPM: Environmental Chemistry Laboratory wasia/EC/Toxicity_ : Environmental Chemistry Laboratory Instruction ... Argentina Genotoxicity, ... Biosensor is used for the determination

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  • Determination of soils' and sediments' toxicity with Heterocypris incongruens assay


    EPM: Environmental Chemistry

    Laboratory Instruction

    Gdask, 2015/2016

  • Determination of soils' and sediments' toxicity with Heterocypris incongruens assay


    Passing requirements: it is obligatory to fully participate in the exercise, no late-coming will

    be considered as an excuse. During the laboratory each student will catch 3-5 living

    organisms (depending on Tutors instructions) and measure their length, these values will be

    checked by Tutor. Error > 20% will be considered as failure to pass the laboratory. After the

    classes students will receive xls file with toxicological data for real soil/sediment sample

    and concluding on possible reason of ecotoxicological will have to be done as a part of

    laboratory report. The individual report (in pdf form) must be given to the Tutor within 7 days

    after the classes, also within 1 week the written tests will be performed individually for each

    student. The theoretical definitions below are only introductory guidelines, more detailed

    information will be presented at the beginning of laboratory classes. Delay in delivering the

    report or writing the tests > 14 days will be considered as not passing the laboratory.

    1. Theoretical background

    Sources and environmental fate of pollutants

    Environment is a very complex system divided into biotic (living) and abiotic (non-

    living) part, among which there is a continuous exchange of matter and energy. These

    processes should remain in balance called homeostasis. The sensitive balance between biotic and abiotic part may be disrupt by chemicals introduced into environment, which are derived

    from natural or anthropogenic sources. Through the years of evolution plant and animal

    organisms have developed number of defense mechanisms against different natural pollution.

    Champions in this race are plants, which have developed many defense mechanisms to

    survive due to the lack of mobility. Changes under the influence of anthropogenic stress can

    have negative effects on the organisms inhabiting a given environment. Every day water, soil

    and air are attacked by hazardous chemicals coming from different industrial branches,

    some of them undergo any legal regulations because these substances have not yet been

    identified or studied. Fig. 1. shows the distribution of pollution in terms of their origin and

    due to the legal regulation. Bold examples of sources of pollutants are classified as a natural

    environment pollution group, as can be seen in Fig. 1., they are a minority among the

    pollutants of anthropogenic origin. Virtually all industries cause environmental pollution, but

    some of them are especially famous for significant impact. Among them one can mention:

    petrochemical industry,

    mining of precious metals and stones,


    lead battery industry,

    industrial discharges and/or municipal.

    Other manifestations of human activity such as: motorization, housing, agriculture,

    sewage and municipal wastes are also not without significance. Negative effects can be cause

    by both organic and inorganic compounds, those that are synthesized by man deliberately or

    formed as a by-product, such as: PAHs (polycyclic aromatic hydrocarbons), PCBs

    (polychlorinated biphenyls), organochlorine pesticides OCPs, BPA (bisphenol A), PCDDs

    (polychlorinated dibenzo - paradioxins), PCDFs (polychlorinated dibenzofurans), PBDEs

    (polybrominated diphenyl ethers), NOx,(mixture of nitrogen oxides NO and NO2) SOx

    (mixture of sulfur oxides SO2 and SO3) and heavy metals. Tab. 1. summarizes information

    concerning compounds considered to be most environmentally burdensome. Part of these

    hazardous compounds has been removed from use years ago, but as may be noted by

    analyzing the data from Tab. 1. their content in the environment is still considerable.

  • Determination of soils' and sediments' toxicity with Heterocypris incongruens assay


    Chemical compounds characterized by toxic properties, that easily undergo

    bioaccumulation processes, are resistant to biodegradation and have high transport potential

    are called persistent organic pollutants (POPs). Due to lipophilic and hydrophobic nature of

    POPs, they accumulate mainly in sediments and soils. Half - life of POPs may vary from

    years or even decades in soils/sediments in contrast to several days half - life in atmosphere.

    More than a decade ago, scientists recognized the problem which may create POPs therefore,

    those compounds become a main topic of the Stockholm Convention on Persistent Organic

    Pollutants signed in 2001.

    Environmental pollution does not respect boundaries and under favorable conditions it

    may be transmitted over long distances and travel throughout the biosphere. Study of the Polar Regions show that pollutants may be transmitted over long distance. Polar Regions and

    its specific environment are ideal to study transport of pollutants, because all the sources of

    emission are distant urban areas.





















    Fig. 1. Classification of environmental pollution due to the their sources and legal regulations

  • Determination of soils' and sediments' toxicity with Heterocypris incongruens assay


    Tab. 1. Information about groups of chemicals showing special environmental arduousness.

    Group of



    Concentrations Sample type Toxic effect

    PAHs 0.69-32.73 [ng/m3] Air sample, Brazil,


    Genotoxicity, carcinogenesis,

    teratogenicity, cytotoxicity,


    PCBs 0.0039-0.0365 [ng/g]

    Soil and water

    samples, river Chao,


    Endocrine disrupting properties,

    cancerogenesis, immunotoxicity,

    hemotoxicity, infertility

    PBDEs 6.3-26050 [ng/L] River water samples,

    river Aire, UK Endocrine disrupting properties

    PBA 82-292 [ng/L] River samples,

    Aisonas river, Greece

    Endocrine disrupting properties,


    PCDD 162-3450 [pg/g] Soil samples, Hong

    Kong, China

    Immunotoxicity, neurotoxicity,

    hepatoxicity, mutagenic


    PCDF 3-213 [pg/g] Soil samples, Hong

    Kong, China mutagenic, cancerogenesis

    OCPs 5-180 [ng/g] Soil samples,

    Shouguang, China

    Endocrine disrupting properties,

    immunotoxicity, neurotoxicity,

    infertility, cancerogenesis

    NOx 110-150 [g/m3]

    Seville metropolita,

    Iberian Peninsula, Spain Acute toxicity, cytotoxicity,

    cytostatic, genotoxicity

    SOx Causes lipid peroxidation processes in organ tissues



    Hg LOD-90 [mg/kg]

    River s and lakes in

    the world

    Acute toxicity, nephrotoxicity,

    cardiotoxicity, cytotoxicity

    Cu 1.33-924.54 [mg/kg]

    Cd LOD-81.79 [mg/kg]

    As LOD-83 [mg/kg]

    Pb LOD-5778.1


    In the scientific terminology any movement of pollutants on a global scale is known as

    long-range transportation. Pollutants may be transferred by different environmental

    components like water and air or by living organisms. Water and air act as a transport

    medium, however transport by living organisms it strongly depends on the migratory species.

    In the environment chemical compounds undergo a number of processes depending on

    their physicochemical properties. Hydrophilic substances remain dissolved in water and

    hydrophobic substances accumulate in soil or/and sediment. Chemicals may be partially

    bioaccumulated by living organisms. Bioaccumulation is a phenomenon in which living

    organisms uptake substances from the environment, when consumption and the amount of

    xenobiotic in different tissues become greater, than the ability of the body to its removal.

    Bioaccumulation is a general term and applies to accumulation of pollutants from the soil,

    water and air. More specific term is bioconcentration, which refers only to accumulation of

    pollutants dissolved in water. Bioconcentration is usually defined by the ratio of chemical

    concentration in organism to concentration in the water and is described by the

    bioconcentration factor (BCF) shown in eq. (1).

    (1) enviromentintoxinofionoConcentrat


  • Determination of soils' and sediments' toxicity with Heterocypris incongruens assay


    Another term that often appears in the literature is biomagnification. In this case,

    accumulation of toxic substances increases along the food chain. Chemical substance,

    depending on the physicochemical properties may be treated by the body in din two ways.

    Hydrophilic compounds are filtered by the kidneys and excreted in the urine, and those

    characterizing by lipophilic properties are metabolized by the liver, from where they are

    excreted in the urine or faeces. Chemical compounds that hav