Abstract:

Vitellogenin is an egg-yolk precursor protein expressed in females of nearly all oviparous species and is commonly used to detect estrogenic effects in male fish. Vitellogenin antibodies were validated for each of the collected fish species and will be analyzed used appropriate antibodies. The vitellogenin assays-concentrations will be reported as ng/ml of plasma.

Suggested Citation:

Wetzel, Dana. 2016. Plasma vitellogenin levels of collected fish species, DeSoto Canyon, August 16-29, 2013. Distributed by: GRIIDC, Harte Research Institute, Texas A&M University–Corpus Christi. doi:10.7266/N71C1TWG

Purpose:

Assessment of vitellogenin levels in plasma to determine liklihood of endocrine disruption.

Data Parameters and Units:

Plate (ID), Assay (ID), Curve Fit Type [linear regression, R-squared], Well Positions (ID), Sample (ID), Raw Data, Curve Fit, Final Results, Sample (ID), Sex (M,F), Species (common name) [Red Snapper, Golden tilefish], Vitellogenin Concentration Rep 1 to Rep 3 (ng/mL), Vitellogenin Concentration Average (ng/mL), Vitellogenin Concentration Standard Deviation, Vitellogenin Concentration % CV coefficient variation of absorbance (%). The following is a list of stations where samples in the dataset were collected from (Station ID, Westbound Extent, Northbound Extent, Southbound Extent, Eastbound Extent): WB-SL10-40, -88.52829, 29.12272, 29.08907, -88.52213; WB-HE-265, -90.30623, 28.23532, 28.22594, -90.28127; WB-SL16-150, -90.00278, 28.3727, 28.3387, -89.3387; WB-SL12-100, -89.35517, 28.3347, 28.32427, -89.33911; WB-SL11-150, -88.37047, 29.03165, 29.02626, -88.3441; WB-SL5-100, -85.26157, 28.3284, 28.32743, -85.22738; WB-SL4-40, -84.25941, 28.07729, 28.05337, -84.24252; The dataset file titled " Vitellogen 2013 Sample Locations" contains information about where each sample in the dataset was collected from and is organized by Plate ID. The following is a description of each field in the file: Fish Sample ID (unique identifier that corresponds to the Sample ID fields in the other dataset files), Station ID (station where sample was collected), Westbound Extent (western bounding extent of station (DD)), Northbound Extent (northern bounding extent of station (DD)), Southbound Extent (southern bounding extent of station (DD)), Eastbound Extent (eastern bounding extent of station (DD)), Center Point Lat (latitudinal center point of station bounding extent(DD)), Center Point Lon (longitudinal center point of station bounding extent (DD)).

Methods:

Sample Collection: plasma - Collect 4 mL whole blood into a green top lithium heparin tube containing aprotinin. - Invert the tube 5 times and allow to clot on ice for 30 minutes. - Spin at 2700 rpm for 15 minutes. - Remove supernatant and place in green top cryovial containing aprotinin. - Store in -20 freezer or in LN2 Sample Processing: Samples are run according to manufacturer's protocol

Instruments:

DS2 Automated ELISA instrument by Dynex Technologies Vitellogenin Rainbow Trout ELISA by Biosense Laboratories

Error Analysis:

Standard curves and no substrate blanks are run with each plate Samples are run in triplicate Sample %CV's are calculated and outliers are removed such that %CV is less than 20%

Individual Files: