# Weekly Rearing Room Schedule As of May 13, 2002

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:
   * Carefully remove ‘S-weight’ from top of egg sheets
   * Using forceps, carefully remove the sheets from the lid of the E/L containers
   * Distribute egg sheets evenly between six (6) ½-pint jars and close jars with solid cap and ring
   * Place jars in 50°F incubator in holding room
   * Replace egg sheets on lids of E/L containers – 4 sheets per lid
   * 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:
   * Fill sugar-water vial with 10% fructose solution
   * Invert vial(s) by placing Petri dish on top; hold together; and invert in one smooth motion
   * Carefully set one inverted sugar-water vial on top center of each E/L container
   * Carefully remove and refill or replace any vials that are less than 1/3 full
   * 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:
   * Using a small camel hair brush, infest each diet cup with 3-4 larvae
   * Place the larvae on the underside of the lid
   * Place lid on cup and push into place, making sure it is fully seated in cup
   * 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:
   * Gently harvest PTB pupae from cups
   * Place pupae from about 3 trays into each E/L container
   * Place E/L containers on shelf in holding room
   * Place 1 or 2 egg sheets on top of container with ‘S’-weights
   * Place sugar-water vial on top center of lid
This work is in the public domain in the United States because it is a work of the United States federal government (see 17 U.S.C. 105).
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