Chapter 25
PROTOCOL
Mollusca
Subprotocol 1
Gonads of Slugs (Pulmonata)
Materials
- Dissecting microscope and dissecting tray
Equipment
- Scissors
- Forceps
- Pasteur pipettes
- Plastic container (6 em deep)
- Culture vessels
- Glass bottle
- Chiarandini's physiological solution (see Appendix 1)
Solutions
- Snail organ culture medium (see Appendix 1)
- Slug Arion subfuscus
Animals
- Filter paper
Miscellaneous
- Cotton wool
- Solid CO 2
Procedure
1. Isolate slugs individually several days before culture into a sterile glass Pretreatment
container whose bottom is covered with sterile filter paper wet with a
sterile solution of penicillin G (100,000 IU1200 ml), streptomycin
(5 mg1200 ml), and polymyxin B (50,000 IU1200 ml).
Without anesthesia, slugs will secrete a considerable amount of mucus Anesthesia
by strong contractions, and this is unfavorable for the dissection of the
cephalic organs. Bailey (1969) developed the following method to anesthetize slugs.
1. Crush solid CO2 and place it under cotton wool, in a layer 1 em deep,
in a plastic container.
2. Place the slugs on moist filter paper on the cotton. The time taken for
anesthesia is variable.