Embryotoxicity of Great Lakes Lake Trout Extracts to Developing Rainbow Trout.

Metadata:

• Identification_Information
• Data_Quality_Information
• Entity_and_Attribute_Information
• Distribution_Information
• Metadata_Reference_Information

Identification_Information:

Citation:

Citation_Information:

Originator: Peggy J. Wright, Donald E. Tillitt

Originator:

U.S. Geological Survey, Biological Resources Division, Columbia Environmental Research Center

Publication_Date: 1999

Title:

Embryotoxicity of Great Lakes Lake Trout Extracts to Developing Rainbow Trout.

Publication_Information:

Publication_Place: Columbia, Missouri

Publisher:

U.S. Geological Survey, Biological Resources Division, Columbia Environmental Research Center

Description:

Abstract:

Planar halogenated hydrocarbons (PHHs), such as polychlorinated dibenzo-p-dioxins, dibenzofurans, and biphenyls are present in aquatic systems, and are known to produce adverse effects in fish. This study investigated the embryotoxicity of PHH mixtures through the nanoinjection of environmental extracts into newly fertilized eggs from two strains of rainbow trout. Organic extracts were obtained from whole adult lake trout collected from Lake Michigan in 1988 and Lake Superior in 1994. The graded doses of the final extracts used for injections were quantified as 2,3,7,8-tetrachlorodibenzo-p-dioxin toxic equivalents (TEQs) based on the concentrations of dioxins, furans and non-o-PCBs in each, and as equivalent amounts found in the eggs of the original lake trout (eggEQ). Total TEQs in the lake trout were 14.7 pg TEQ/g in the Lake Michigan sample and 7.3 pg TEQ/g in the Lake Superior sample. The extract of the Lake Michigan lake trout was embryotoxic to rainbow trout; LD50 values were 35 eggEQ (15-90, 95% F.L.) in the Arlee strain and 14 eggEQ (5-99, 95% F.L.) in the Erwin strain of rainbow trout. The L.D50 values of the Lake Michigan extract in either of these strains of rainbow trout fall within the actual range of TCDD LD50 values based on TEQs. This indicates that an additive model of toxicity is appropriate to quantify PHHs in relation to early life stage mortality in fish. Gross lesions characteristic of exposure to PHHs (i.e. yolk-sac edema, craniofacial deformities, and hemorrhaging) increased in a dose-related manner. The lowest observable adverse effect concentrations (LOAEC) for these gross lesions and cumulative mortalities suggests that current concentrations of PHHs in lake trout from Lake Michigan are above a threshold for adverse effects and these compounds may have implications on the lack of recruitment in certain Great Lakes lake trout populations.

Purpose:

The objectives of the study were to extract and quantitate the PHHs present in Lake Michigan lake trout tissue; determine the embryotoxic effects of this extract toward rainbow trout; determine if an additive model of toxicity of PHHs adquately describes the embryotoxic effects of this chemical mixture; and estimate the hazard these chemicals may represent toward lake trout populations in the Great Lakes.

Time_Period_of_Content:

Time_Period_Information:

Multiple_Dates/Times:

Single_Date/Time:

Calendar_Date: 1988

Calendar_Date: 1994

Currentness_Reference: observed (fish collection dates)

Status:

Progress: Complete

Maintenance_and_Update_Frequency: None planned

Keywords:

Theme:

Theme_Keyword_Thesaurus: None

Theme_Keyword: additivity

Theme_Keyword: chemical mixtures

Theme_Keyword: developmental toxicity

Theme_Keyword: polychlorinated biphenyls

Theme_Keyword: PCBs

Theme_Keyword: salmonids

Theme_Keyword: 2,3,7,8-TCDD

Place:

Place_Keyword_Thesaurus: None

Place_Keyword: Lake Superior

Place_Keyword: Lake Michigan

Taxonomy:

Taxonomic_Keywords: rainbow trout

Taxonomic_Coverage:

Specific_Taxonomic_Information:

Kingdom: Animal

Division-Phylum: Chordata

Class: Actinopterygii

Order: Salmoniformes

Family: Salmonidae

Genus: Oncorhynchus

Species: Oncorhynchus mykiss

Applicable_Common_Names: rainbow trout

Access_Constraints: None

Use_Constraints: None

Point_of_Contact:

Contact_Information:

Contact_Organization_Primary:

Contact_Organization:

U.S. Geological Survey, Biological Resources
Division, Columbia Environmental Research Center

Contact_Person: Donald Tillitt

Contact_Address:

Address_Type: mailing and physical address

Address: 4200 New Haven Road

City: Columbia

State_or_Province: Missouri

Postal_Code: 65201

Contact_Voice_Telephone: (573) 876-1886

Contact_Facsimile_Telephone: (573) 876-1896

Contact_Electronic_Mail_Address: Donald_Tillitt@usgs.gov

Data_Set_Credit:

John Meadows, Dennis Schroeder, and Mark Alexander for their technical assistance with extract preparation, and Kathy Echols, Paul Peterman and Robert Gale for chemical analyses. Susannah Cantrell, Pam Alt, and Diane Nicks for assistance in the lab.

Cross_Reference:

Citation_Information:

Originator: Peggy J. Wright, Donald E. Tillitt

Publication_Date: 1999

Title:

Embryotoxicity of Great Lakes lake trout extracts to developing rainbow trout.

Series_Information:

Series_Name: Aquatic Toxicology

Issue_Identification: 47

Other_Citation_Details: pp. 77-92.

Data_Quality_Information:

Attribute_Accuracy:

Attribute_Accuracy_Report:

Mortality data and pathological lesion data were analyzed by probit, corrected for control responses.

Logical_Consistency_Report: not applicable

Completeness_Report: unknown

Lineage:

Methodology:

Methodology_Type: Lab

Methodology_Identifier:

Methodology_Keyword_Thesaurus: None

Methodology_Keyword: chemicals

Methodology_Description:

Triolein (95% purity, Sigma) was used as the vehicle for the extracts. The triolein was filtered using disposable sterile syringe filters (25 mm, 20 micrometers, cellulose acetate membrane), and stored in 1 ml vials. 2,3,7,8-TCDD (99% purity, Dow Chemical, gift) was stored in hexane at room temperature until the appropriate dose in triolein was made. Extracts were obtained from processed lake trout tissue.

Methodology_Type: Lab

Methodology_Identifier:

Methodology_Keyword_Thesaurus: None

Methodology_Keyword: extraction and clean-up of lake trout tissue

Methodology_Description:

Archived lake trout tissue collected in 1988 from Sheboygan, WI on Lake Michigan was used for the extraction of the complex environmental mixture. The lake trout was part of the National Contaminant Biomonitoring Program (NCBP) (Schmitt et al., 1990). Extraction and clean-up procedures were followed according to Meadows et al. (1993). Briefly, a total of 4.832 kg of Lake Michigan lake trout tissue (whole fish) wsa ground and mixed with four times the sample wet weight of Na2SO4 (sodium sulfate) in order to dehydrate the tissue. The homogenized lake trout tissue was then extracted in large, 4-cm i.d. glass extraction columns with methylene chloride (Ch2Cl2). The extracted lipid was dialyzed against 80:20 hexaneCH2Cl2 utilizing polyethylene membranes. The sample was then subjected to a two-step clean-up on sulfuric acid-impregnated silica gel and potassium silicate, and final clean-up by adsorption chromatography. The clean-up procedures effectively enrich the target compounds (i.e. PHHs), while removing lipids and many non-target compounds such as oxygenated organochlorine pesticides and labile compounds. The final sample was then subject to high-performance gel permeation chromatography (HPGPC). Dosing solutions were prepared by evaporation of the extract in methylene chloride, and then dissolution of the residues into triolein. The stock solution of the extract (4.832 kg/ml) was diluted to obtain the appropriate amounts of each dose. Whole lake trout collected in 1994 from Devils Island Shoal, Lake Superior (Station 22, NCBP) was used as a negative control for the Lake Michigan injection experiment. Lake Superior lake trout reproduce successfully in the wild, and are considered relatively 'clean' when compared to Lake Michigan lake trout. A total of 4.8 kg of Lake Superior lake trout tissue (whole fish) was processed. The extraction and clean-up procedure was identical to that used for the Lake Michigan tissue, as was the dosing solution preparation.

Methodology_Citation:

Citation_Information:

Originator: Schmitt, C.J., Zajicek, J.L., Peterman, P.H.

Publication_Date: 1990

Title:

National Contaminant Biomonitoring Program: residues of organochlorine chemicals in U.S. freshwater fish, 1976-1984.

Series_Information:

Series_Name: Arch. Environ. Contam. Toxicol.

Issue_Identification: 19

Other_Citation_Details: pp. 748-781.

Methodology_Citation:

Citation_Information:

Originator:

Meadows, J., Tillitt, D.E., Huckins, J., Schroeder, D.

Publication_Date: 1993

Title:

Large-scale dialysis of sample lipids using a semipermeable membrane device.

Series_Information:

Series_Name: Chemosphere

Issue_Identification: 26

Other_Citation_Details: pp. 1993-2006.

Methodology_Type: Lab

Methodology_Identifier:

Methodology_Keyword_Thesaurus: None

Methodology_Keyword: analytical preparation methods

Methodology_Description:

Triplicate aliquots (25 g) of both the Lake Michigan lake trout sample and the Lake Superior lake trout sample were prepared for analysis. Samples were stored frozen at -20 degrees C in their original containers until sample processing began. Two additional quality control samples were also prepared and spiked: a procedural blank and a positive control carp sample (Saginaw Bay, Michigan). Approximately 25-g aliquots of the samples were blended with three times their weight of anhydrous sodium sulfate, and homogenized for determination of PHH concentrations in each. Each sample was spiked with 5 ng of 13 C-labeled non-o-PCBs (#77, 126 and 169) using 50 ml of MSC Standard 91W-2, and column-extracted with CH2Cl2. All extracts were then treated by a two stage reactive clean-up; employing first, a sulfuric acid silica gel/potassium silicate (SASG/KS) column, and second, a column of sulfuric acid silica gel/potassium silicatae/silica gel (SASG/KS/SG) The extracts were further purified using HPGPC. PCDDs, PCDFs, and PCBs were separated on PX-21-activated carbon dispersed on C18 HPLC packing material and analyzed as previously described (Feltz et al, 1995). Briefly, HPLC-carbon loadings were determined by GC/ECD analyses of total PCBs in each extract. The analytes were then separated by HPLC-C, isolating four fractions: fraction 1 bulk, bulk and di-o-PCB congeners; fraction 2, mono-o-PCB congeners; fraction 3, non-o-PCB congeners; and fraction 4, PCDDs/PCDFs. Mono-o-PCB congeners were determined by GC/ECD, while non-o-PCBs, PCDDs and PCDFs were determined by gas chromatography/high-resolution mass spectrometry (GC/HRMS) as described by Peterman et al. (1966). Dioxin toxic equivalents (TEQs) in the samples were calculated with toxic equivalency factors (TEFs) calculated with toxic equivalency factors (TEFs) developed for early life stage mortality in rainbow trout where available (Walker et al, 1991); Zabel et al., 1995), or TEFs from the PLHC-1 bioassay (Tillitt and Cantrell, 1992) and an additive model of toxicity.

Methodology_Citation:

Citation_Information:

Originator:

Feltz, K.P.; Tillitt, D.E.; Gale, R.W.; Peterman, P.H.

Publication_Date: 1995

Title:

Automated HPLC fractionation of PCDDs and PCDFs and planar and nonplanar PCBs on C18-dispersed PX-21carbon.

Series_Information:

Series_Name: Environ. Sci. Technol.

Issue_Identification: 29

Other_Citation_Details: pp. 709-718.

Methodology_Citation:

Citation_Information:

Originator:

Walker, M.K., Spitsbergen, J.M., Olson, J. R., Peterson, R. E.

Publication_Date: 1991

Title:

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) toxicity during early life stage development of lake trout Salvelinus namaycush.

Series_Information:

Series_Name: Can. J. Fish. Aquat. Sci.

Issue_Identification: 48

Other_Citation_Details: pp. 875-883.

Methodology_Citation:

Citation_Information:

Originator: Zabel, E.W., Cook, P.M., Peterson, R.E.

Publication_Date: 1995

Title:

Toxic equivalency factors of polychlorinated dibenzo-p-dioxin, dibenzo-furan, and biphenyl congeners based on early life stage mortality in rainbow trout (Oncorhynchus mykiss).

Series_Information:

Series_Name: Aquati. Toxicol.

Issue_Identification: 31

Other_Citation_Details: pp. 315-328.

Methodology_Citation:

Citation_Information:

Originator: Tillitt, D.E., Cantrell, S.M.

Publication_Date: 1992

Title:

Planar Halogenated Hydrocarbon (PHH) structure Activity Relationship in a Teleost (PLHC-1) Cell Line.

Series_Information:

Series_Name: Society of Environmental Toxicology and Chemistry

Issue_Identification: 13th Annual Meeting Cincinnati, Ohio

Publication_Information:

Publication_Place: Pensacola, Florida

Publisher: SETAC Press

Methodology_Type: Lab

Methodology_Identifier:

Methodology_Keyword_Thesaurus: None

Methodology_Keyword: rainbow trout fish eggs

Methodology_Description:

Rainbow trout eggs were obtained from Ennis National Fish Hatchery, US Fish and Wildlife Service, located in Ennis, Montana. Unfertilized rainbow trout eggs (pooled from at least three females) and milt (pooled from at least two males) arrived packed in ice (~4 degrees C), and were fertilized after several hours of slow warming to within 1 degree Centigrade of the incubator water temperature (10 + or - 1 degree Centigrade). After water-hardening, eggs were placed in preformed agarose plates in preparation for injection.

Methodology_Type: Lab

Methodology_Identifier:

Methodology_Keyword_Thesaurus: None

Methodology_Keyword:

Injection of Lake Michigan and Lake Superior extracts

Methodology_Description:

Three dose-response experiments using the Lake Michigan lake trout extract were conducted, two with Arlee strain rainbow trout eggs, and one with Erwin strain eggs. Additionally, two dose-response studies were conducted withthe Lake Superior lake trout extract, both using Erwin strain eggs. Doses, measured as egg-equivalents (eggEQ) were based on gram tissue/gram egg values normalized to the lipid content of lake trout eggs (Mac et al., 1985; Herbert and Keenleyside, 1995). Doses used were control (triolein), 0.02, 0.10, 0.20, 1.0, 2.0, 4.0, 10.0 and 20.0 eggEQ per gram of egg were prepared and used. Injections were conducted using newly fertilized eggs, and were completed within 48 hours post-fertilization. The injection method allows small volumes of liquid to be delivered precisely with glass micropipettes, a regulated gas pressure system, and a digital control device which are described in detail in Walker et al. (1996), The injection volume delivered into the yolk of each egg (~50 nl, or 0.1% of egg volume) was quanitified by measuring the size of the triolein droplet which forms at the tip of the needle during injection.

Methodology_Citation:

Methodology_Citation:

Citation_Information:

Originator: Herbert, C.E., Keenleyside, K. A.

Publication_Date: 1995

Title:

To normalize or not to normalize? Fat is the question.

Series_Information:

Series_Name: Environ. Toxicol. Chem

Issue_Identification: 14

Other_Citation_Details: pp. 801-807.

Methodology_Citation:

Citation_Information:

Originator:

Walker, M.K., Zabel, E.W., Akerman, G., Balk, L., Wright, P., Tillitt, D.E.

Publication_Date: 1996

Title:

Fish egg injection as an alternative exposure route for early life stage toxicity studies. Description of two unique methods.

Larger_Work_Citation:

Citation_Information:

Originator: Ostrander, G. (Ed.)

Publication_Date: 1996

Title: Techniques in Aquatic Toxicology

Edition: vol. 4

Publication_Information:

Publication_Place: unknown

Publisher: CRC Press Lewis Publishers

Other_Citation_Details: pp. 41-77.

Methodology_Type: Field

Methodology_Identifier:

Methodology_Keyword_Thesaurus: None

Methodology_Keyword: maintenance of rainbow trout eggs and fry

Methodology_Description:

Eggs were reared in a vertical flow incubator, with water temperature held at 10 degrees (+/- 1 degree). The egg fertilization rate was consistently greater than 85%, and unfertilized eggs were discarded and not considered in data analysis. Mortality was recorded daily through swim-up for all experiments. The incidence of gross physical abnormalities was quantified on day 41 (post fertilization) for all of the Lake Michigan and Lake Superior experiments. Live fry in each dosing group on day 41 were examined individually for three main categories of gross pathological lesions: yolk-sac edema, hemorrhaging, and craniofacial deformities.

Process_Step:

Process_Description:

Mortality data and gross pathological lesions in the two different strains of rainbow trout eggs (Arlee and Erwin) were analyzed separately due to their different relative sensitivities to TCDD and related compounds (Zabel et al., 1995). Egg mortality was included in calculations of LD50 values because dose-related mortality was evident in the egg stage of development, unlike in previous TCDD and TCDF studies with rainbow trout embryos (Walker et al., 1992). Mortality data and pathological lesion data were analyzed by probit, corrected for control responses (SAS, 1988). Probit models were tested for goodness-of-fit (P values > 0.05). The no observable adverse effect level (NOAEL) and the lowest observable adverse effect level (LOAEL) for mortality data and pathological lesion data were analyzed using Dunnett's test (P < 0.05; SAS, 1988) comparing treatment and control groups.

Process_Date: unknown

Entity_and_Attribute_Information:

Overview_Description:

Entity_and_Attribute_Overview:

Entity - Rainbow trout; Associated attributes - embryotoxicity

Entity_and_Attribute_Detail_Citation: unknown

Distribution_Information:

Distributor:

Contact_Information:

Contact_Organization_Primary:

Contact_Organization:

U.S. Geological Survey, Biological Resources
Division, Columbia Environmental Research Center

Contact_Person: Christopher Henke

Contact_Position: Webmaster

Contact_Address:

Address_Type: mailing and physical address

Address: 4200 New Haven Rd

City: Columbia

State_or_Province: MO

Postal_Code: 65201

Contact_Voice_Telephone: 573-875-5399

Contact_Facsimile_Telephone: 573-876-1896

Contact_Electronic_Mail_Address: chris_henke@usgs.gov

Distribution_Liability:

Although these data have been processed successfully on a computer system at the U.S. Geological Survey, no warranty expressed or implied is made regarding the accuracy or utility of the data on any other system or for general or scientific purposes, nor shall the act of distribution constitute any such warranty. This disclaimer applies both to individual use of the data and aggregate use with other data. It is strongly recommended that these data are directly acquired from a U.S. Geological Survey server, and not indirectly through other sources which may have changed the data in some way. It is also strongly recommended that careful attention be paid to the contents of the metadata file associated with these data. The U.S. Geological Survey shall not be held liable for improper or incorrect use of the data described and/or contained herein.

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Metadata_Reference_Information:

Metadata_Date: 200004

Metadata_Contact:

Contact_Information:

Contact_Organization_Primary:

Contact_Organization:

Raytheon Information Technology and Scientific
Services (ITSS)

Contact_Person: Cheryl Solomon

Contact_Position: Ecosystem Coordinator

Contact_Address:

Address_Type: Mailing and Physical Address

Address: 4500 Forbes Boulevard

City: Lanham

State_or_Province: MD

Postal_Code: 20706

Country: USA

Contact_Voice_Telephone: 301 794-3049

Contact_Facsimile_Telephone: 301 794-3164

Contact_Electronic_Mail_Address: solomon@gcmd.nasa.gov

Metadata_Standard_Name:

NBII Content Standard for National Biological Information Infrastructure Metadata

Metadata_Standard_Version: December 1995

Metadata_Access_Constraints: None

Metadata_Use_Constraints: None