Weekly Rearing Room Schedule As of May 13, 2002

by United States Department of Agriculture

INSECTARY: USDA, ARS, SJVASC, Parlier, CA

SOP Rearing Procedure: PTB (Rev. October 2007)

Peach Twig Borer, Anarsia lineatella Zeller; Lepidoptera: Gelechiidae

Origin: 2004; Fresno County; Parlier, CA; U.C. Davis, Kearney Agricultural Center; Laboratory Colony

Diet/Culture: Lima Bean Agar Diet in 1-ounce Cups (See Appendix)

Tasks Performed: Weekly

Day	Procedure
Monday	Harvest/Replace Egg Sheets (→ 50°F); Pull Diet Trays for Tomorrow
Tuesday	Infest Diet Cups; Check Sugar-water Vials
Wednesday	Harvest Pupae & Set up E/L Containers
Thursday	Harvest/Replace Egg Sheets (→ 50°F); Discard Oldest E/L Containers
Friday	Move Eggs from 50°F Incubator to Shelf in Rm. #1 (80°F); Check Sugar-water Vials

Harvest/Replace Egg Sheets (E/S):

Materials:

• PTB E/S with eggs
• PTB replacement E/S
• Forceps
• ‘S’-weights
• ½ pint jars with solid lids and rings

Procedure – performed on shelves in holding room:

1. Carefully remove ‘S-weight’ from top of egg sheets
2. Using forceps, carefully remove the sheets from the lid of the E/L containers
3. Distribute egg sheets evenly between six (6) ½-pint jars and close jars with solid cap and ring
4. Place jars in 50°F incubator in holding room
5. Replace egg sheets on lids of E/L containers – 4 sheets per lid
6. Put one ‘S-weight’ on top of each egg sheet

Refill/Replace Sugar-water Vials (10% Fructose):

Materials:

• 7-dram clear plastic vials with bridal veil material held tightly over top with two rubber bands
• 10% fructose solution
• Petri® dish top or bottom

Procedure – performed on shelves in holding room and over sink in handling room:

1. Fill sugar-water vial with 10% fructose solution
2. Invert vial(s) by placing Petri dish on top; hold together; and invert in one smooth motion
3. Carefully set one inverted sugar-water vial on top center of each E/L container
4. Carefully remove and refill or replace any vials that are less than 1/3 full
5. To avoid spills, lift vials straight up, smoothly – but not too slowly
   Infest cups – Set up new colony:

Materials:

• PTB neonate larvae
• PTB cups (1 oz.) with fresh diet
• Small camel hair brush
• Paper lids

Procedure – performed under fumehood:

1. Using a small camel hair brush, infest each diet cup with 3-4 larvae
2. Place the larvae on the underside of the lid
3. Place lid on cup and push into place, making sure it is fully seated in cup
4. Label trays and place on shelf in holding room for development

Set up Egg Layers (E/L):

Materials:

• PTB pupae
• Oviposition containers with lids with bridal veil centers
• Larval forceps

Procedure – performed under fumehood:

1. Gently harvest PTB pupae from cups
2. Place pupae from about 3 trays into each E/L container
3. Place E/L containers on shelf in holding room
4. Place 1 or 2 egg sheets on top of container with ‘S’-weights
5. Place sugar-water vial on top center of lid