Selection of Biological Methods for Industrial Effluents Assessments

1. Selection of Biological Methods
for
Industrial Effluents Assessments
Rubinger. Carla Ferreira1, Leão. Mônica Maria Diniz, Paulinyi. Helene Maria
Universidade Federal de Minas Gerais
Departamento de Engenharia Sanitária e Ambiental
Programa de Pós graduação em Saneamento, Meio Ambiente e Recursos Hídricos
Belo Horizonte – Minas Gerais, Brasil.
E-mail: carlarubinger@ufmg.br

2. Sustainable Development
Production Quality
Environmental Quality
Cleaner Production

3. Water Quality Standard
Effluent Quality Standard
Physical – Chemicals Methods
Complexity of Wastewater
Effects on Biota

4. GOALS
This study aimed to identify existing biological methods, bioassays of
acute and chronic toxicity, seeking their comparison and their
applicability to the assessment of quality of treatment applied to
industrial effluents and to understand its advantages and limitations.

5. RESULTS
Therefore we performed a survey of the main methods of toxicity
tests used in the analysis of industrial effluents.
Identifying existing bioassays for the evaluation of acute and chronic
toxicity of industrial wastewater, so this has led to comparisons
between the main methods used for the analysis of chronic toxicity
of effluent from its applicability, contributing to the compilation of
data for environmental monitoring.

6. BIOLOGICAL METHODS
Biological methods or bioassays are tests that use living organisms
to produce information on the development and changes, the
effect of acute or chronic condition that a particular chemical may
exert on living matter.

7. Biological methods can be divided into three groups for analysis:
 Microbiological: those who engage primarily in detection of
pathogens;
 Limnology: provide information on the development and
changes in long-term biotic community in aquatic ecosystems;
 Ecotoxicological: reveal acute or chronic effects produced by
chemicals or compounds on living organisms.

8. Ecotoxicological studies generally include:
 Study of emissions and inputs of pollutants in the abiotic
environment, distribution and destiny in the various
compartments.
 Study of entry and fate of pollutants in biological chains and their
forms of transfer via the food chain as food;
 Qualitative and quantitative study of the toxicological effects of
pollutants on the ecosystem with consequences for man.

9. ECOTOXICOLOGICAL TESTS
Toxicity tests to assess the effects of test
species, are the exposure of aquatic
organisms representative of the
environment to various concentrations of
one or more substances or environmental
factors, for a certain period of time. These
tests are called bioassays or biological
assays.

10. AQUATIC TOXICITY TESTS
Duration
Method of adding the test solution
Purpose

11. SELECTION OF TEST METHODS
Bioassays with indicators of short life cycle
Several beings
Easy handling
Speed in performing a test
Potential effects of synthetic compounds in the biota.

12. LEGISLATION AND STANDARD RELEVANCE
 Brazilian Environmental Law
 Internationally Regulation

13. LEGISLATION AND STANDARD RELEVANCE
Toxicity tests are standardized by national and international
associations and standards organizations, such as:
 Brazilian Technical Standardization Association (ABNT),
 French Association for Standardization (AFNOR),
 American Society for Testing and Materials (ASTM),
 American Association Works with Water (AWWA),
 German Institute for Standardization (DIN),
 International Organization for Standardization (ISO),
 Organization for Economic Cooperation and Development (OECD).

14. ECOTOXICOLOGICAL IDENTIFIED TESTS

15. ECOTOXICOLOGICAL IDENTIFIED TESTS
Test-organisms used in bioassays of toxicity test
Acanthocyclops robustus Danio rerio Pimephales promelas
Artemia salina Daphnia ambigua Placopecten magellanicus
Asellus aquaticus Daphnia magna Poecilia reticulata
Biomphalaria tenagophila Daphnia pulex Pseudokirchneriella subcapitata
Brachionus calyciflorus Daphnia similis Saccharomyces cerevisiae
Brachionus plicatilis Daphnia spinulata Salmonella typhimurium
Ceriodaphnia dubia Enterobacter aerogenes Scenedesmus quadricauda
Ceriodaphnia silvestrii Euglena gracilis Scenedesmus subspicatus
Chironomus riparius Gammarus pulex Selanastrum capricornutum
Chlamydomonas reinhardi Hyalella curvispina Tetrahymena pyriformis
Clorella sp Hydra oligactis Tetrahymena thermophila
Cnesterodon
Lemna minor Thamnocephalus platyrus
decemmaculatum
Coliforms and bacterian flake Onchorynchus mykiss Vibrio fisheri
Crassostrea rhizophorae Oryzias latipes Vorticella sp

16. TOXICITY TO ALGAE
Algae in an aquatic ecosystem are part of the group of primary producers, the words are
the base of the aquatic life cycle. Participate in maintaining the balance of these
environments through their roles in nutrient cycling of biogeochemical cycles, particularly
in the carbon, oxygen, nitrogen, phosphorus and silicon. Also part of the diet of several
other types of organisms, both aquatic and terrestrial.
Growth inhibition test for algae Chlorella vulgaris
Test method of chronic toxicity test with algae(Chorophyceae)
Test of inhibition of growth and fluorescence Scenedesmus subspicatus

17. TOXICITY TO ALGAE
Lemna minor
Pseudokirchneriella subcapitata
Clorella vulgaris

18. TOXICITY TO MICROCRUSTACEOUS
Microcrustaceuos herein are plankton of freshwater and saltwater. They act as primary
consumers among metazoan, feeding by filtration of particulate organic material, mainly
algae.
Inhibition test swimming ability of Daphnia magna
Acute toxicity test for Daphnia similis
Test of chronic toxicity to Daphnia similis
Test Evaluation of Chronic Toxicity to Ceriodaphnia dubia

19. TOXICITY TO MICROCRUSTACEOUS
Daphnia magna
Daphnia similis Daphnia pulex

20. TOXICITY TO MICROCRUSTACEOUS
Ceriodaphnia dubia Ceriodaphnia quadrangula

21. TOXICITY TO FISH
Fishes are vertebrates, aquatic freshwater and saltwater representing consumers of
various orders (primary, secondary and tertiary). Tests with fish as test-organisms are
generally intended to understand how a toxicant affects agencies greater degree of
structure, such as vertebrates and how these changes can affect more complex cycles
of bioconcentration, bioaccumulation, biomagnification along the aquatic ecosystem.
Often fish are used as environmental indicators because they are widely studied
organisms, because they develop well in laboratory cultures and are easily observed.
Evaluation tests of acute toxicity to fish
Test evaluation of chronic toxicity to fish
Assessment Test Bioconcentration in Fish

22. TOXICITY TO FISH
Cyprinodon variegatus
Brachydanio rerio
Danio rerio
Cnesterodon decemmaculatum
Pimephales promelas

23. TOXICITY TO BACTERIA
Bacteria used as test-organisms can be grouped with decomposers in the food chain of
aquatic ecosystems. They act in the degradation of organic deposits resulting from dead
organisms or their metabolites into simpler substances so that they can be easily
reassimilate bodies producers and also consumers of such filter feeders. For this reason
become good indicators of environmental quality has that an environmental pollutant,
even at low concentrations, can be identified by these bodies and answers in a short
period of time.
Test of inhibition of bacterial bioluminescence
Cytotoxicity test (Ames test)
Genotoxicity testing (Umu test)

24. TOXICITY TO BACTERIA
Vibrio fischeri Salmonella typhimurium

25. ANALYSIS OF TEST METHODS
Routine monitoring of effluents
Research in industrial effluents
Regular uniformity of procedures
Precision data for the validation of results

26. Comparison between the test methods most commonly used in toxicological evaluation of industrial
effluents.
Test – organism Type Test duration Variable response Expression results
Unicellular Green
Acute 72h Specific growth rate of average ECNO
Algae
Acute 48h Mobility (ability nadatória) in 24h and 48h EC50
Acute 48h Immobility of 50% at the end of 48 h EC50
Microcrustacian
Daphnia sp ECNO, Cex(50, 20,
Chronic 21 days Number of offspring, mortality among the parental
10%)
Microcrustacian Paternogenica number of offspring, survival of adult
Chronic 7 days ECNO
Ceriodaphnia SP females
Acute 96h 100% mortality of the lowest effective concentration LC50
Chronic 7 - 10days Appearance and abnormal LC50
Phish
Chronic 14 - 28days Mortality and abnormal behavior and appearance ECNO
Chronic 10 – 14days Shock and survival, appearance and abnormal behavior ECNO
Acute 15min - 2h Decrease bioluminescence ECNO
ECNO, Cex(50, 20,
Chronic 2 – 3days Reducing the number of colonies, synthesis of histamine
10%)
Bacteria
Reducing the number of colonies, synthesis of β - ECNO, Cex(50, 20,
Chronic 30h
galactose 10%)

27. CONCLUSIONS
The methods of biological assays are preventive action procedures, and essential
in the estimation of environmental impacts, were used by the authorities.
There is no consensus on what would be the best type of test organisms for a
particular effluent, but what set of tests should be developed to ensure that the
components and byproducts of the degradation of effluents cause the least
adverse impact to aquatic biota and consequently to humans.

28. CONCLUSIONS
The test methods for toxicity tests are acute or chronic, to an effluent as a whole,
they generally follow several standard protocols that include a description of use
and culture of the species recommended as test organisms. There is clearly a
tendency to use test-organisms which are regulated and standardized over the
use of native species.
Among the test-organisms as the most frequent search terms are highlighted in
the cladoceran Daphnia magna and Ceriodaphnia dubia, followed by the
bioluminescent bacterium Vibrio fischeri. No less significant is the use of algae as
Pseudokirchneriella subcapitata and fish species such as fathead minnows.

29. CONCLUSIONS
Recent studies show changes in standardized test methods as an attempt to
increase the degree of security in the face of the same composition of the biota
of specific local in which you want to develop particular study.
Because they are standardized tests and described internationally longer than
the others, the tests using cladocerans as daphnideos are more widespread in
research and regulations aimed at examining the toxicity of effluents and
chemicals complex.
The studies suggest that the toxicological evaluation of aquatic ecosystems by
the combined or isolated from an environmental pollutant is recommended that
biological testing is done in different trophic levels of the ecosystem.

30. CONCLUSIONS
Regarding the response time of a test-organism to the presence or absence of
toxicant action of an effluent or chemical compound, it is recommended the use
of tests with bacteria, because they generally have a duration of 30 minutes to 2
days only, for sensitivity, the toxicity of industrial effluents and compounds
chemical complex there is a definition of pattern or individual species described
as more or less sensitive to the action of pollutants.
The tests are preferred to chronic acute tests, since they describe situations of
damage deeper in the body test and consequently the impact that this toxicant
can cause the biocenose that this pollutant is inserted.