Our project on the Atlantic cod, Gadus morhua, received funding in the Fall of 1996 and I am reporting the progress made toward its goals in its second quarter.

Progress Outline:

1. Existing-Data analysis: Existing plus new sample analysis continues.
2. Serum Sample Analysis: We are proceeding with our study of cod serum proteins.
3. New Tissue Gathering: Necessary contacts have been successfully used.
4. Equipment Acquisition: Data archival equipment was obtained using other funding.
5. Personnel Identification: A work-study student was found to help with lab work.
6. New Sample Processing: New cod tissue samples have revealed useful information.
7. Publication: Hartling, Pereira and Kunkel (1997) is in press.

1. Existing-Data analysis: The existing data analysis is being refined and will be exhibited as WWW pages over the summer.
2. Serum Sample Analysis: Several serum samples of male serum have been run on the new spherical cellulose gel filtration media columns. The elution pattern for male serum did not resolve enough free column room for the vitellogenin (Vg) and lipovitellin (Lv) we expect to detect in the female samples. This suggests that we should use our previous Agarose A1.5 media approach instead.
3. New Tissue Gathering: We have obtained 15 samples of whole cod ovaries from egg laden cod females with the cooperation of long-line cod fishermen from the Chatham MA harbor. These have been frozen and are being used to develop a purification scheme for cod lipovitellin. Thus we have successfully formed a relationship with a series of fishermen who will provide us with all the material we need in the lipovitellin purification and antibody production phase of our project. They promise to also be useful in the serum sampling phase of the project since they have agreed to have us on their boats to collect fresh samples of serum from cod at sea.
4. Equipment Acquisition: We have obtained a CD writer (Pinnacle Micro 4x4) which we will be using to store images of tissue sections and specimens for archiving as well as rapid retrieval.
5. Personnel Identification: A work-study student has been identified who will be helping with lab organization and cleanup.
6. New Sample Processing: We have done initial experiments on heat denaturation of cod ovarian extracts which suggest that cod Lv, like flounder Lv, is heat stable, and thus, heat denaturation to remove contaminant proteins will be useful in the developing purification procedure.
7. Publication: Hartling, Pereira and Kunkel (1997) is in press in J Exp. Zool., the first paper contributing to Ruth Hartlings CMER funded dissertation is in press. This paper on winter flounder has significance to our present project on cod because it describes the technique of purification of flounder Lv which we are trying to extend to cod Lv. It also describes the immunological assay developed for flounder which we also will use as a model to apply to cod.

In general we seem to be on track to having a purified Lv and an antiserum for use in the Fall season of cod fishing.

Respectfully submitted,

Joseph G. Kunkel

jgk/hs