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ANALISIS EKOTOKSIKOLOGI Oleh Sudrajat FMIPA UNMUL 2010. Pengujian Toksisitas. Uji Toksisitas Akut Short time frame exposure (96h) “kill ‘em and count ‘em” LC 50 , TLM (median tolerance limit EC 50 (effective concentration) Uji Toksisitas Kronis
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ANALISIS EKOTOKSIKOLOGI Oleh Sudrajat FMIPA UNMUL 2010
Pengujian Toksisitas Uji Toksisitas Akut • Short time frame exposure (96h) • “kill ‘em and count ‘em” • LC50, TLM (median tolerance limit • EC50 (effective concentration) Uji Toksisitas Kronis • Longer time frame exposure (1 week to 3 years) • Endpoints are reproduction (brood size) physiology, behavior, biochemistry • More ecologically relevant
Uji Toksisitas Akut • Battery, consisting of four tropic levels • Vibrio fischeri luminescent bacteria test • Bacteria • Selenastrum capricornutum growth inhibition test • Algae • Daphnia pulex acute toxicity test • Invertebrate (waterflea) • Poecilia reticulata acute toxicity test • Vertebrate (fish)
Toxicity units • Toxicity units • Determine LC/EC50 value from dilution series tested • Toxicity units (TUa) = 100/LC or EC50 • Key to classification system: 0-1 Limited to not acutely toxic 1-2 Negligibly acutely toxic 2-10 Mildly acutely toxic 10-100 Acutely toxic >100 Highly acutely toxic
Applications • Surface and Groundwater • Drinking water • Mine water • Sediment pore water • Industrial effluents/discharges • Soil leachates • Extracts of solid wastes • Waste dump leachates • Stormwater run-off • Chemicals and products
Treated sewage effluent • Plant receiving domestic and industrial discharges • Discharged into a stream where very little if any dilution takes place • Water upstream and downstream of discharge tested
Biological And Microbiological Assessment And Treatment Of Water Present Standards (30) • Bactericidal and algaecidal efficacy of swimming pool chemicals • Determination of BOD • Water quality - Enumeration of microorganisms Vibrio cholerae, bacteriophages, E. coli • Biocides for use in industrial cooling water • Detection and enumeration of Legionella • Guidance on validation of microbiological method; Uncertainty of Measurement
Future Standards Legionnaires' disease: Risk Assessment; Control in water systems ADOPTING international standards for DEEEP methodology • Vibrio fischeri (Luminescent bacteria test) (ISO 11348-3:2007, second edition). • Algal Growth Inhibition Tests (ISO 8692 2004) • Daphniapulex acute toxicity test (US EPA 2002). • Fish, acute toxicity test (OECD Guideline for testing of chemicals - 203, 1992) Voting on future ISO standards for toxicity and Microbiotest kits internationally
Acute vs. Chronic Toxicity Tests Can broadly classify toxicity tests based on length of exposure • Acute Toxicity test • Drop dead testing • Time = 2 days (invertebrates) to 4 d. (fish) • LD50 • LC50 • TLm (median tolerance dose) • EC50 (effective concentration) • Lose equilibrium, sit on bottom “ecologically” dead • Not very ecologically relevent but quick, relatively cheap (but still ~$700-1,200 per test)
Acute vs chronic toxicity testing (con’t) • Chronic toxicity testing • Growth, reproduction • More ecologically relevant data but takes longer, more expensive • Shows effect at much lower dose • Test requires much more “baby-sitting”
Acute Testing - theory • Population of organisms has normally distributed resistance to toxicants acute toxicity test designed to identify mean response • Regulations allow 5% of species to be impacted • Most tests only use 2-3 species (up to 6) not really enough to protect 95% of all species!
Acute Toxicity Test Organisms • Use of test species based on • Lab hardiness • Common • Known life cycle • Cheap • Short-lived