Rearing of Trichoplusia ni

OVIPOSITION CAGES
DISINFECTING EGGS
PINTO BEAN LARVAL DIET
LARVAL-PUPAL CUPS
DISINFECTING PROCEEDURE FOR PUPAE
SEXING PUPAE
SOURCE OF READING MATERIAL
AGING LARVAL INSTARS

OVIPOSITION CAGES
 1/4 "  hardware cloth is used to make a  cylindar 12" h  x  8" diameter . Cover top of cylindar with hardware cloth and also fsaran wrap to confine insects.  A hole 1" in diameter is cut in the top for access to sugar water supply.  A 200 ml. pyrex beaker , filled with 10% sugar water solution , covered with organdy cloth and secured with a rubberband, is inverted and placed over hole in the top of the cage. This serves as the food source for the moths.  Wax paper is wrapped around the outside of the cage, and secured with tape. The moths will oviposition between the hardware cloth and onto the wax paper. The cage is placed on a tray over a petri dish, containing a layer of moistened Vermiculite. About 70-95 mixed male and female pupae are placed on the Vermiculite and allowed to emerge. Keep the vermiculite moist as this helps adults to emerge with proper wing expansion.

DISINFECTING EGGS

1. Disinfect egg sheet with 10 % clorox solution.
2 .Rinse again with distilled water. Use squirt bottles to gently run solutions down over eggs.
3. Sheets are then placed between  clean towels to dry.

LARVAL DIET

1.Cut up ovipsition sheet into squares which have 12 - 15 eggs / square.
2. Scotch tape( one square to the LID of each diet cup containing Pinto bean diet. Larvae will hatch and string down onto the diet in the bottom of the cup, which contains about 3/8 " of media.
3. Place Pinto Beans and water in blender and chop briefly - 2 sec.
     Pinto Beans (Ground)     300 gms.
     Water, distilled      700 ml.
4. Add next 6 dry ingredients and chop briefly for 1 seconds, 4-5 times
     Brewers Yeast ( fortified )    64.0 gms.
     Ascorbic Acid        6.0 gms.
     Methyl paraben        4.0 gms.
     Sorbic Acid         2.0 gms.
     Aureomycin, 14.1 % active      1.0 gm.
     Propionic Acid, Na salt       1.5 gms.
5. Cook agar and water until thickened, not boiling.
     Agar          27.0 gms.
     Water, distilled      550.0 mls.
6. Add cooked agar to ingredients in blender and mix thoroughly. Remove bl  ender cover, let media cool 2 minutes before adding vitamin solution.
7. Add vitamin solution and formaldehyde to blender ingredients. Stir with spoon carefully and then blend thoroughly.
     Vitamin Mix        10.0 gms.
     Formaldehyde          4.0 mls.
8. Immediately, while hot, transfer media to appropriate containers.
9. Allow media to cool 3 hours or longer until excess moisture has evaporated from surface   before adding larvae and capping.

DIET / PUPATION CUPS
Pupation cups are held in a growth chamber which has a photo period of 16 : 8  L/D hrs., and a temperature of 78 degrees fahrenheit.
Cups are held for approximately  2 1/2  weeks, at which time most larvae have pupated and need to be collected. Pull pupae out, webbing and all.

DISINFECTING PROCEEDURE FOR PUPAE
 Place pupae with webbing in a solution of 10% clorox and stir around gently with  your fingers for 60 seconds. This separates the pupae from the webbing and also disinfects pupae. Rinse pupae carefully  under room temperature tap water, about 60 seconds,until clorox smell is gone. Place rinsed pupae on several layers of paper towelling. Pull rest of webbing from pupae. Pupae are now ready to sex and/or use for a new mating cage.

SEXING PUPAE

Hold the pupae in good light, count down the segments below the segment covered by the wing buds.
If the forth segment (or 8th segment on A-P axis) is marked, notched, split or therwise not completely intact -> Female; if entire -> Male
Male with a very noticable slit, very close to anal opening, usally on top of perturberance.

 

 

 

 

 

 

SOURCE OF REARING MATERIAL

--VS 508 Sweetheart Hot Food Cups  1000 / Case    $73.45
--LS8 Sweetheart Lids                       2000 / Case    $102.75
           Seneca Bar & Restaurant Supply 20 E. Castle St. Geneva, N. Y.  14456
           Phone #  315-789-2422

 --Propionic acid (Sodium salt, 99%) Aldrich Chemical Co., Inc., Milwaukee, WI  53233

 --Formaldehyde (certified ACS-F-79, 37% w/w solution)
   Fisher Scientific Co.,  Fair Lawn, NJ  07410

 --All other ingredients: Bio-Serv Inc., P.O. Box 450, Frenchtown,  NJ

Procedure modified by Andre Levy from protocol by Kathy Poole, NYSAES, Cornell
 

Trichoplusia ni  -  Aging Instar Stages

Larval instar is established by measuring width of head capsule, with the aid of a microscope and a ocular with micrometer.
   1st Instar     0.2-0.24 mm
   2nd Instar    0.4-0.48 mm
   3rd Instar    0.72-0.8 mm
   4th Instar    1.12-1.2 mm
   5th Instar    1.8-2 mm

    General description

Eggs that are on the verge of hatching assume a black coloration, corresponding to color of 1st instar head capsule.
Early Instar stage -  head and body are even in size.
Late Instar stage -   head size stays constant but body size of larva increases about 4 times, therefore head looks comparatively small.
Pre-molt stage  -  Very distinctive , easy to pick out .  Head and body are separated by a whitish 'swollen neck'  area.  Therefore the head again looks comparatively small.