A Fish Two Era Poisonous quality Test Point by point Audit Paper Endocrine Disruptor Strategies Approval Subcommittee De.


79 views
Uploaded on:
Category: Food / Beverages
Description
Point by point Review Paper: A Fish Two Generation Toxicity Test Detailed Review Paper. WORK PERFORMED BY:andSpringbornSmithers Laboratories LLC for the United States Environmental ProtectionAgency EPA CONTRACT NUMBER 68-W-01-023. Routines USED IN THIS ANALYSIS. On-line Literature Search
Transcripts
Slide 1

A Fish Two Generation Toxicity Test Detailed Review Paper Endocrine Disruptor Methods Validation Subcommittee December 2002 Leslie Touart

Slide 2

Detailed Review Paper: A Fish Two Generation Toxicity Test Detailed Review Paper WORK PERFORMED BY: and SpringbornSmithers Laboratories LLC for the benefit of the United States Environmental Protection Agency EPA CONTRACT NUMBER 68-W-01-023

Slide 3

METHODS USED IN THIS ANALYSIS On-line Literature Search "Exchange" On-Line look with database Biosis Previews Aquatic Science and Fisheries Abstracts Endocrine disruptor screening strategies for fathead minnow, zebrafish, medaka, sheepshead minnow Key Words "estrogen* or testosteron* or endocrin* or antiandrogen* or androgen* or hormon* or thyroxin* or * thyroid * technique, convention and so forth… 7453 records were refined down to 601 records

Slide 4

METHODS USED IN THIS ANALYSIS Cont. Interviews With The Following Experts Tom Hutchinson Reynaldo Patino Taisen Iguchi Alf Lundgren Dan G. Cyr Nancy Denslow

Slide 5

METHODS USED IN THIS ANALYSIS Cont. Outer/Internal Peer Review Dr. Dave Hinton – Duke Unv. USA Dr. John Sumpter – Brunell Unv. UK Dr. Gary Thorgaard - Univ. of Wash. USA EPA Technical Experts

Slide 6

OVERVIEW AND SCIENTIFIC BASIS Evidence exists that EDCs influence sexual separation, improvement, and propagation in Fish life cycle test techniques have been institutionalized for quite a long time Existing strategies don\'t evaluate transgenerational impacts and need applicable biochemical, morphological, and behavioral endpoints The proposed Fish Two Generation Test addresses these EDC important oversights

Slide 7

Test Species Fathead minnow ( Pimephales promelas ) Medaka ( Oryzias latipes ) Zebrafish ( Danio rerio ) Sheepshead minnow ( Cyprinodon variegatus) little size at development simplicity of society support costs nonconcurrent spawners

Slide 8

Fathead Minnow Family Cyprinidae 35 to 75 mm length Extensive sea-going poisonous quality in USA era time around 4 months sexually dimorphic females produce 50 to 250 incipient organisms for every bring forth

Slide 9

Strengths sufficiently large to gather singular blood plasma tests Distinct optional sex attributes in both genders Large recorded administrative database Many research centers are acquainted with society and testing Spawn on a substrate High treatment rate Indigenous to North America Weaknesses Relatively long life cycle Relatively high variability in fertility Relative size of the fish requires more space for society and testing Intersex condition is less as often as possible watched contrasted with different fishes. Genome inadequately portrayed Fathead Minnow

Slide 10

Medaka Family Adrianichthyidae indigenous to Japan, Taiwan, and southeastern Asia Generation interim of 2 to 3 months sexually dimorphic 25 mm to 50 mm length females produce 10 to 30 eggs for every bring forth evaluated to be more than 500 developed strains Genetically Engineered/Inbred Strains in Toxicity Testing

Slide 11

Strengths Relatively short life cycle Relatively little fish, making society and testing conceivable in littler space Female sex decided amid fetus stage versus male sex decided after portal Sex-connected shading strain Weaknesses Smaller size diminishes singular blood test volumes contrasted with fathead minnow Less particular optional sex attributes Regulatory information base less broad contrasted with fathead minnow. Constrained use in transient tests in the U.S.A. Medaka

Slide 12

Zebrafish Family Cyprinidae Native to East India and Burma 4 cm to 5 cm long Extensive sea-going harmfulness in Europe Difficult to sex zebrafish Sexual development in 10 to 12 weeks 150 to 400 eggs for each female Development of transgenic zebrafish

Slide 13

Strengths Short life cycle Small fish, making society and testing conceivable in littler spaces Male fish experience an androgynous stage as adolescents Widely utilized as a part of other medicinal and hereditary research Frequently utilized as a part of Europe for administrative purposes Transgenic angle progressively accessible Anticipated that whole genome will be sequenced soon. Shortcomings Small size makes singular blood plasma tests not likely Minimal auxiliary sex qualities Limited US administrative information base Limited testing knowledge in the US Zebrafish

Slide 14

Sheepshead Minnow Family Cyprinodonitidae Native to Atlantic and Gulf of Mexico estuaries 35 mm to 50 mm length Tolerates wide ranges in temperature (0 to 40ºC) and saltiness (0.1 ppt to 149 ppt) Sexually dimorphic Sexual development in 60 days Low variability in fruitfulness Large authentic administrative database

Slide 15

Strengths Very short life cycle (<60days to sexual development), seawater expenses might be balanced by shorter presentation times for testing Males sufficiently substantial for individual blood plasma tests Distinct sexual dimorphism Relatively low variability in fertility Relatively huge chronicled administrative database Many research centers are acquainted with society and testing Relatively little fish making society and testing conceivable in littler space Weaknesses Estuarine/marine species, saltiness of 15 to 30 ppt prescribed, be that as it may, lower saltiness might be conceivable (5 ppt) Culture requires countless to deliver enough eggs in a 24-hr period to start an existence cycle test Limited data on conceptive endocrinology Small size makes singular blood plasma tests not likely Sheepshead Minnow

Slide 16

Routes of Administration of Chemical Exposure Aqueous Dietary exposures Direct infusion systems Intravascular intraperitoneal

Slide 17

Measurement Endpoints Growth and Morphological Alterations Gonadosomatic Index Histology Techniques Sexual Differentiation Secondary Sex Characteristics Measures of Reproductive Performance Fecundity Gamete Viability Changes in Spawning Behavior Biochemical Measures Vitellogenin Induction Tissue Steroid Concentrations Thyroid hormones

Slide 18

MEASUREMENT OF BIOCHEMICAL ENDPOINTS Sex Steroids in Tissues Estrogens/Androgens/Progestins Radioimmunoassay (RIA) Enzyme-connected Immunosorbent Assay (ELISA) Liquid/Gas Chromatography with Mass Selective Detection (LC/GC-MS)

Slide 19

Measurement of Vitellogenin Indirect Quantification of Vitellogenin Protein Alkaline-labile Phosphate Assay Direct Quantification of Vitellogenin Protein RIA Enzyme-connected Immunosorbent Assay Antibody-catch Antigen-catch Direct Enzyme-connected Immunosorbent Assay Sandwich Enzyme-connected Immunosorbent Assay Quantifying Vitellogenin mRNA Ribonuclease Protection Assay (RPA) Quantitative Reverse Transcription-Polymerase Chain Reaction (QRT-PCR) Mass spectrometry (MS)

Slide 20

CANDIDATE PROTOCOLS Partial Life-Cycle Test {Adult (P) to Juvenile (F1)} 2) Full Life-Cycle Test {Egg (P) to Juvenile (F1)} 3) Multi-Generation Test {Egg (P) to Juvenile (F2)} 4) Two Generation Test {Adult (P) to Juvenile (F2)}

Slide 21

Partial Life-Cycle Test {Adult (P) to Juvenile (F1)} A fractional life-cycle lethality test, which uncovered P grown-up, sexually develop fish and the early life cycle of F1 fish, ( can be utilized to gauge the NOEC) A pre-introduction regenerative assessment is led on the P fish. Organic endpoints assessed: P pre-introduction, optional sexual attributes and fruitfulness/proliferation (e.g., eggs/female) P post-presentation, survival, auxiliary sexual qualities, fertility/generation (e.g., eggs/female), GSI, histopathology, and protein and sex steroid biomarkers (e.g., VTG) F1 bring forth achievement, survival, development (length and weight)

Slide 22

Full Life-Cycle Test {Egg (P) to Juvenile (F1)} A full life cycle test created (Benoit 1981-fathead minnows ) and (Hansen et al. 1978 - sheepshead minnow). Started with prepared eggs (P) and the fish are consistently uncovered through regenerative development, took after appraisal of the early improvement of the F1 era. Natural endpoints assessed: P developing life time-to-seal, incubating achievement, larval survival and length, weight of diminished fish, survival, optional sexual attributes, fruitfulness/multiplication (e.g., eggs/female), development. F1 bring forth achievement, survival, development (length and weight)

Slide 23

Multi-Generation Test {Egg (P) to Juvenile (F2)} Multi-era lethality test, uncovered all life-phases of two eras of fish Test is started with eggs and two full eras of fish are uncovered amid the test (can gauge the NOEC) Biological endpoints assessed: C P and F1 bring forth achievement, survival, development (length and weight), time-to-development, sex proportion, auxiliary sexual attributes, fruitfulness/multiplication (e.g., eggs/female), and protein and sex steroid biomarkers (e.g., VTG). F2 bring forth achievement, survival and development.

Slide 24

Two Generation Test {Adult (P) to Juvenile (F2)} Two era life-cycle poisonous quality test, uncovered the grown-up P, full F1generation,and measures F2 reasonability Variables including the course of events of the test, the quantity of fish required in the test, and getting endpoints, for example, Vtg plasma levels will be connected with the diverse species. Natural endpoints assessed: P Survival, optional sex qualities, conceptive conduct, bringing forth action, fruitfulness, treatment achievement F1 bring forth achievement, survival, development (length and weight), time-to-development, sex proportion, auxiliary sexual attributes, fertility/proliferation (e.g., eggs/female), and protein and sex steroid biomarkers (e.g., VTG). F2 bring forth achievement, survival and development.

Slide 25

Timeline forTwo-Generation Protocol with the Fathead Minnow - 3 0 3 18 21 25 Time (weeks)

Slide 26

Significant Data Gaps • Male-particular impacts of estrogen agonists other than VTG enlistment. • The impacts of hostile to estrogens • The impacts of androgen agonists and opponents • Baseline information for thyroid hormone levels in test species. • The impacts of thyroid hormone agonists (or thyroid incitement) on proliferation. Pu

Recommended
View more...