Materials for Caterpillar Salivary Gland Removal: Procedure A
Dissecting microscope
Small dissecting scissors
Small forceps
4-8 Dissecting pins for each dissection (can be reused)
Parafilm wax
Parafin wax (optional)
Petri dishes (9cm x 2cm, 1 per dissecting group)
Petri dishes (5cm x 1.5cm, 1 per dissecting group)
Styrofoam container filled with ice
1000 mL beaker
1.5 mL microcentrifuge tubes or sealable containers of similar volume
Distilled water

Materials in Addition to the Above for Caterpillar Salivary Gland Removal: Procedure B
Tetracycline (Optional antibiotic - available through Caroline Biological Supply and/or local livestock feed stores depending on your area)
Hydrogen peroxide (Household variety, 3%)
Liquid dish soap
Three 500 mL beakers
Medicine cups lined with tissue paper
2 Parafilm wax strips (for holding down caterpillars)

Planting Procedures (Tomato plants should be planted approx. 1 month before experiment)
1. Thoroughly dampen soil and place one to three seeds on the surface of the soil in each pot. If you place more than one seed in a pot,
allow for approximately 3 inches of spacing between seeds, in a pot that is large enough to maintain more than 1 month old tomato plant.
(Usually 3 seeds are sown in case some seeds do not germinate. Unwanted plants are removed in early development).
2. Cover the seeds lightly with a coat of dampened soil.
3. Place in growth area.
Note: Tomato plants are light loving plants. Optimal growth conditions are in a greenhouse with at least 14 hours of daylight and temperatures
of approximately 24-28 °C. However, plants placed on a window sill with bright light, and/or supplemented with indoor lighting should work
fine. If this project is performed during a normal calendar school year, and the window sill is the only option, try to plan the experiment during
the longest daylight hours of a particular semester.
During the first week, water as often as necessary to keep the soil damp. After the seedlings are visible, the watering schedule should be based
on soil dampness and the appearance of plant vigor. When the soil surface is relatively dry (before plant wilting), water the tomato plants again
thoroughly. As the plants become larger or stressed from mechanical wounding caused by the student researchers or caterpillar feeding on the
plants, the watering schedule will typically need to be increased. Fertilize the tomatoes once a week after the seedlings are visible
(4.5g Nitrogen/m2, N:P:K=15:30:15, American Plant Food Company, Creve Coeur, MO) following the manufacturer’s guidelines. Tomato
plants respond well to a variety of fertilizers and particularly phosphate fertilizer. At four weeks the tomato plants should have at least three fully
expanded compound leaves, be approximately 0.25 m tall, and can be utilized in the experiments (see tomato anatomy web site
http://www-plb.ucdavis.edu/cou.3.rses/plb105/Students/Tomato/Tomhome.html).

Caterpillar Rearing Procedure
Raising Neonates for Experiments and Dissection
1) Set out plastic medicine cups filled with approx. 1 teaspoon caterpillar diet, and have lids ready.
2) Lay out paper towel; this will help you spot potential runaway neonates on the table.
3) Open container of neonates and place on paper towel (including lid, as it is probably also covered in neonates).
4) Pick up one caterpillar at a time by gently touching it with the paint brush.
5) Deposit neonate onto diet, one per cup, either by tapping brush gently on rim of cup or, if caterpillar has let out a line of silk,
by lowering it onto diet and allowing it to touch diet (it will grab onto the food by itself). Make sure each caterpillar is touching
the diet, or it may never find it and starve.
6) Cover medicine cup with lid.
7) Place trays of caterpillars into a warm room (24-28 o C), preferably under a light timed for 15 hrs of light, 9 hrs dark, for approx.
two weeks (or until they’ve reached the sixth instar, and are approx. 2.5 cm long).
Tomato fruit worm caterpillars, H. zea can be obtained from the Insectary at the Department of Entomology NC State University (http://www.cals.ncsu.edu/entomology/insectary/homepg.html) for a few cents per larvae and can be shipped to your school.
The Entomology Department at NC State also sells artificial diets that the caterpillars can be reared on.. Insects can also be purchased
from other Entomology Departments, or the USDA, as well as biological supply houses such as Carolina Biological Supply Company.
One important note is that H. zea caterpillars are cannibalistic, so there is a need to maintain individuals in separate rearing containers such
as clear small (4-cm diameter) medicine-cups, small clear empty deli containers, or airtight sandwich bags if long term rearing of the
caterpillars is desirable. There is no need to punch holes into the tops of these containers, as there is ample oxygen for the caterpillar metabolic
needs (this is a common misconception spurred from punching holes in jar lids from childhood).
Two options for rearing caterpillars include using artificial diet, in which case you will need a few grams of diet per caterpillar (follow the
instructions provided by the manufacturer), or you can also make your own caterpillar artificial diet by readily available grocery store ingredients;
see Goodman et al. (2001) for details. Caterpillars can also be reared on tomato leaves. If you decide to use tomato leaves, grow extra plants,
extra large so that there is ample leaf material to feed the caterpillars. Leaf material not eaten by the caterpillar will need to be discarded after 5 days
and replaced with fresh leaves. If the leaves become too dry or moldy they will need to be replaced sooner. Often adding a little tissue or paper
towel under the leaf and caterpillar will soak up excessive moisture from the humid leaves and will prevent mold, and also prevent small caterpillars
from drowning in the excess moisture. An airtight container will help keep the leaves from drying out.
The caterpillars grow optimally in a warm environment with temperatures of approximately 24-28 °C and 15 hours of daylight. The insects do not
need direct light to grow, but the light maintains the caterpillar’s circadian rhythms. If for some reason the caterpillars grow too rapidly in relation
to the tomato plants, you can chill the caterpillars in the refrigerator for a couple of days and the growth will be slowed down. Similarly the
caterpillars will grow very slowly if they are in an air-conditioned room (sometimes too slow to survive, if over several days). If the classroom
is too cool the caterpillars will not grow very well as insects are ectothermic (traditionally referred to as "cold blooded") and so the environmental
conditions greatly
affect their growth.

The following procedures can be performed by students as in class activities or by teachers before hand.
Dissecting Platform Preparation Procedure
1) Melt strips of parafilm in glass petri dish on a hotplate/microwave until approx _ filled with wax.
2) Remove dish from hotplate/microwave. Let parafilm cool and harden.
Paraffin wax can be substituted for parafilm. Paraffin wax can be melted in a separate container and poured into either glass or plastic petri dishes.

Caterpillar Salivary Gland Removal: Procedure A.
In this procedure, caterpillar salivary glands are removed to make salivary extract and the caterpillars discarded (See figs. 1, 2, and 3 ).
1) Fill 500 mL beaker with ice and add water. Place desired number of caterpillars into ice water; let them cool there for at least 15 min.
(otherwise they start to revive upon dissection!)
2) Fill 1.5mL microcenterfuge tube halfway full w/distilled water and tuck it into ice-filled styrofoam box; this will keep the salivary
glands cool after they have been removed.
3) Fill petri dish _ of the way with distilled water. This facilitates the dissection.
4) Place caterpillar ventral side up on petri dish with wax. Push one pin through caterpillar’s head region and another through its posterior end.
5) Snip off one of the prolegs, usually the middle one on your right (if right handed).
6) Snip carefully and shallowly longitudinally up to the neck so as to cut only cuticle or "skin" (if the digestive tract pops through, you’ve cut
too deep -- discard the caterpillar and start another one).
7) Pin down each side of the cuticle to expose inside of caterpillar ventral area.
8) Carefully remove the salivary glands with forceps by pulling on them near the top of the glands near the head – they break easily, so pull gently!
Salivary glands are long, transparent, thin, tube-like structures that run nearly the length of the caterpillar’s body on each side. There should be one
per side.
9) Place salivary glands into microcentrifuge tube on ice. You may need to shake them a little to get the glands off of the forceps and into the water
10) Check caterpillar to be sure there are no fragments of gland remaining
11) Repeat w/another caterpillar. You will probably have to change the water in the petri dish every 15 caterpillars or so, since it becomes more
cloudy with each dissection.Caterpillar Salivary Gland Removal: Procedure B.
In this procedure, caterpillar salivary glands are removed and caterpillars are kept alive for use in experiments (See Fig 4a and 4b).1) Fill 1000 mL
beaker with room temperature tap water. Add 2-3 drops of liquid dishwashing soap(approx. 10mg/L) and enough hydrogen peroxide to make a
0.5% H2O2 solution. This will clean the caterpillars before dissection, as well as render them "unconscious."
2) Fill three 500 mL beakers with distilled water.
3) Optional - In one of the 500 mL beakers, add just 1mg/mL tetracycline. This should turn the water a very pale yellow.
4) Fill petri dish _ full with tetracycline solution (you can just pour straight from the beaker described in step 3).
5) Line bottom of each medicine cup with a small piece of tissue paper, one cup for each caterpillar you plan to dissect.
6) Place caterpillars into hydrogen peroxide mixture (no more than five at a time – this way, you have time to get to all of them before they drown).
7) As soon as they stop moving, remove them and dip them first into one beaker of distilled water, then the second, then finally into the beaker
with the tetracycline solution.
8) Place a caterpillar onto the petri dish ventral side up. Fold 2 pieces of parafilm wax several times to make 2 thin "straps" approx. 4cm long
and 0.5cm wide. Place one strap across the head region of the caterpillar (perpendicular to body) and secure each end of the strap by pinning it
to the wax. Repeat for posterior end of caterpillar. This holds the caterpillar in place.
9) Using the scissors, make as small an incision as possible into its "neck" on the second abdominal segement. The salivary glands may pop
out on their own. The scissors should leave the incision V-shaped, with a triangular flap above it. Fold back the flap.
10) If the glands are not immediately accessible, gently apply a small amount of pressure (using a fingertip or soft forceps) to the caterpillar’s
abdomen to squeeze out the glands.
11) Grasp glands with the forceps and gently slide them back and forth to remove the glands in their entirety.
12) Discard the glands by dabbing them onto a tissue wipe, OR, if also collecting them for salivary extract, deposit them into a microcentrifuge
tube half-filled with distilled water on ice.
13) Using the forceps, fold the triangular flap of cuticle back down and hold it over the incision for at least 30 seconds.
14) Unpin flaps; gently set caterpillar into medicine cup on the tissue paper on his back
15) Repeat. Be efficient with your time – you do not want the other caterpillars to drown.
16) It usually takes 30 minutes – 1 hour for the caterpillars to revive. Approximately 3 - 5 hours after the operation they can be used in experiments.

Mock Surgery:
For experimental control purposes you may desire to use a caterpillar that underwent the same procedure for salivary gland removal but the
salivary glands were left intact. Follow the same procedure for the above but after making the V-shaped incision with the scissors, temporarily
lift back the cuticle ("skin") flap then press it down and hold as directed in the above without removing the salivary glands.
This is the mock dissection.

Salivary Extract Procedure
Salivary glands are used to make a solution to place on mechanically wounded leaves to better simulate herbivory.
1) After putting approximately 10 salivary glands into 1.5 mL microcentrifuge tube, empty contents into small 5 cm petri dish, or similar container.
Add 200 uL distilled water.
2) Place petri dish on ice.
3) While on ice, use bottom of microcentrifuge tube to grind/mash the salivary glands in the water until the glands are broken up and distributed
evenly throughout the water.
4) Use in plant experiments immediately, or if this is not possible, place in freezer (-20oC or -80oC). Avoid thawing and refreezing.

Experimental Procedure A: Simulated Herbivory 1
The mean caterpillar growth rate for caterpillars feeding on plant tissue from mechanically wounded plants is compared to the mean caterpillar
growth rate for caterpillars feeding on plant tissue from non-wounded plants.

1) Wound 10 experimental plants by cutting off the tips of leaves with scissors (on tomato plants approx. 2-3cm). The number of leaves cut
will depend on how large the plants grew. The goal is to damage approx. 20-30% of the leaves. Following good experimental design cut off
the same leaves and same number of leaves for each plant so that they are as identical as possible.
2) Leave 10 tomato plants undamaged as a control.
3) Give plants 4 days to stimulate plant defenses.
4) After 4 days remove leaves from 5 non-wounded tomato plants and divide into 20 medicine cups with one caterpillar neonate per cup
(1st instar caterpillars that just hatched from eggs).
5) Also remove leaves from 5 wounded tomato plants and divide into 20 different medicine cups with one caterpillar neonate per cup.
Following good experimental design, make sure the neonates are of the same age and randomly assigned a treatment.
At this point you should have 40 cups with one neonate per cup. 20 cups contain non-wounded leaf material and 20 cups contain wounded
leaf material. At this stage of development, caterpillar neonates are too small to be weighed accurately so we consider their initial weights "0mg."
However, if you start with larger caterpillars you should weigh them before feeding them the leaf material.
6) Allow caterpillars to feed for 5-6 days. All caterpillars need to be in identical environmental conditions since factors such as temperature
will also influence their growth rates.
7) After feeding for 5-6 days, weigh individual caterpillars and obtain an average weight for each treatment.
8) Remove leaves from the 5 remaining non-wounded tomato plants and replace the leaf material in neonate cups corresponding to that treatment.
9) Remove leaves from the 5 remaining wounded tomato plants and replace the wounded leaf material in neonate cups corresponding to that
treatment.
10) Allow caterpillars to feed for another 5-6 days.
11) After 5-6 days, weigh the caterpillars for a final weight.
12) Determine the caterpillar weight difference before and after feeding and determine average weight gain.

Experimental Procedure B: Simulated Herbivory 2
The mean caterpillar growth rate for caterpillars feeding on plant tissue from mechanically wounded plants treated with salivary extract
are compared to the mean caterpillar growth rate for caterpillars feeding on mechanically wounded plants treated with water (for salivary
extract control). These treatments are also compared to the mean caterpillar growth rate for caterpillars feeding on plant tissue from
non-wounded plants.

1) Wound 10 experimental plants by cutting off the tips of leaves with scissors (on tomato plants approx. 2-3cm). The number of leaves
cut will depend on how large the plants grew. The goal is to damage approx. 20-30% of the leaves. Following good experimental design
cut off the same leaves and same number of leaves for each plant so that they are as identical as possible.
2) Dip the tip (0.5cm) of each wounded leaf in distilled water.
3) Wound 10 experimental plants by cutting off the tips of leaves with scissors (on tomato plants approx. 2-3cm). The number of leaves
cut will depend on how large the plants grew. The goal is to damage approx. 20-30% of the leaves. Following good experimental design
cut off the same leaves and same number of leaves for each plant so that they are as identical as possible.
4) Dip the tip (0.5cm) of each wounded leaf in caterpillar salivary extract solution.
5) Leave 10 tomato plants undamaged as a control.
6) Give plants 4 days to stimulate plant defenses.
7) After 4 days remove leaves from 5 non-wounded tomato plants and divide them into 20 medicine cups with one caterpillar neonate
per cup (1st instar caterpillars that just hatched from eggs).
8) Remove leaves from 5 wounded tomato plants and divide into 20 different medicine cups with one caterpillar neonate per cup.
9) Remove leaves from 5 wounded tomato plants that were dipped in salivary extract and divide into 20 different medicine cups with one
caterpillar neonate per cup.
Following good experimental design, make sure the neonates are of the same age and randomly assigned a treatment.
At this point you should have 60 cups with one neonate per cup. 20 cups contain leaf material from non-wounded plants. 20 cups contain
leaf material from mechanically wounded plants treated with water. 20 cups contain leaf material from plants treated with salivary extract.
At this stage of development, caterpillar neonates are too small to be weighed accurately so we consider their initial weights "0mg."
However, if you start with larger caterpillars you should weigh them before feeding them the leaf material.
10) Allow caterpillars to feed for 5-6 days. All need to be in identical environmental conditions since factors such as temperature will also
influence their growth rates.
11) After feeding for 5-6 days, weigh individual caterpillars and obtain an average weight for each treatment.
12) Remove leaves from the 5 remaining non-wounded tomato plants and replace the leaf material in medicine cups corresponding to
that treatment.
13) Remove leaves from the 5 remaining mechanically wounded tomato plants treated with water and replace the leaf material in medicine
cups corresponding to that treatment.
14) Remove leaves from the 5 remaining mechanically wounded tomato plants treated with salivary extract and replace the leaf in medicine
cups corresponding to that treatment.
15) Allow caterpillars to feed for another 5-6 days.
13) After 5-6 days, weigh the caterpillars for a final weight.
14) Determine the caterpillar weight difference before and after feeding and determine average weight gain.

Experimental Procedure C: Actual Herbivory 1
The mean caterpillar growth rate for caterpillars feeding on plant tissue from plants wounded by caterpillar feeding with intact salivary
glands is compared to the mean caterpillar growth rate for caterpillars feeding on plant tissue from non-wounded plants.Follow Experimental
Procedure A , but replace one of the wounded treatments with the following:
Use caterpillars that are at least 10 days old. Allow them to feed for 24 to 48 hours on the tomato plants.
Remove the caterpillars, wait four days and then feed the tomato leaves to neonates

Experimental Procedure D: Actual Herbivory 2
The mean caterpillar growth rate for caterpillars feeding on plant tissue from plants wounded by caterpillar feeding with intact salivary
glands that underwent the mock dissection are compared to the mean caterpillar growth rate for caterpillars feeding on plant tissue from
plants wounded by caterpillar feeding with removed salivary glands. These treatments are also compared to the mean caterpillar growth
rate for caterpillars feeding on plant tissue from non-wounded plants.
Follow Experimental Procedure B, but replace one of the wounded treatments with caterpillars with intact salivary glands that underwent
the mock dissection. Replace the other wounded treatment with caterpillars with removed salivary glands.

Interpreting the Data (Halpern, 2000)
1) Plot the average caterpillar bodyweight from day 1 (if using neonates this is "0mg"), to day 5 or 6, and a final weight on day 10 or 12.
(See Fig. 5).2) Calculate the mean (X bar) of each treatment. X bar = (X1 + X2…+…Xn)/N
Where X is each weighed caterpillar and N is the total number of caterpillars for that treatment.
3) Calculate the standard deviation (represented by lowercase sigma or "s")

To do this, first find the difference between all of your measurements for a treatment and the mean for that treatment. Square each result
to avoid negative numbers.
(X1 – X bar)2 + (X2 – X bar)2 …+… (Xn – X bar)2
Then divide the result above by the total number of caterpillars for that treatment minus one (N-1).
Finally, take the square root.
4) Calculate the standard error (s.e.). This is the standard deviation divided by the square root of the sample size.
5) Multiply the standard error by 2 (2 s.e.). This allows us to approximate a 95% confidence interval for analysis of our means. Add this
number (2 .s.e) to the mean you got from step 2. This is the "upper 95% confidence limit." Now, Subtract (2 s.e.) from the mean from
step two. This is the "lower 95% confidence limit."
6) Prepare a Dice-Leraas graph (see Halpern, 2000 and Fig 6).
Use a vertical line to represent the range of measurements you obtain for each treatment. The line will connect the points representing the lowest
and highest value within each sample.
Position a horizontal line perpendicular to the range at the point of the mean in each treatment (X bar).
Draw a rectangular box around the mean. The box represents the 95% confidence interval for the mean. The top of the box will be the value of
the "upper 95% confidence limit." The bottom of the box will be the value of the "lower 95% confidence limit." If this is done correctly the box
will be symmetrical around the horizontal line for the mean.
7) Compare the confidence intervals and means. Make a decision whether or not the means are significantly different based on the
following criteria:
A. If the rectangle of just one sample falls within the rectangle denoting the confidence interval of the comparison sample, the means are
not significantly different.
B. If neither rectangle falls within the rectangle denoting the confidence interval of the comparison sample, the means are significantly different.

Experiment Options
The above procedures are examples of experiments you can do. It is also possible to "mix and match" them and compare the effects of
mechanical wounding versus that of actual herbivory. Furthermore you can develop experiments with increasing complexity for the simulated
insect herbivory assays through the application of caterpillar oral secretions to wounded edge of leaves cut with scissors if you choose not to
do the dissections. Slightly squeezing a caterpillar abdomen between your finger tips will cause the caterpillar to regurgitate as a fright
response (see fig 7). This regurgitant can be collected with a fine tipped plastic pipette/capillary tube and stored in the refrigerator for a
few days. After the leaf is wounded this regurgitant can be applied to the wound to simulate caterpillar saliva. Methods of determining
salivary effects on induced plant defenses can be reviewed at Musser et al. (2002 a, b and 2003). These experiments can be further
developed and designed for science fair experiments, or undergraduate/graduate research projects. For example, the amount of wounding
caused on the plant may tested on a scale from no wounding to wounding that is greater than 90% of the total tomato leaves. Or the
experiment may be modified to test the effectiveness of induced plant defenses over time, to determine the number of days after tomato
plant wounding when the stimulated plant defenses are most or least effective. Experiments can also be performed to determine how
fertilizing, daylight conditions, or age of the plant affect induced plant defenses. For additional induced plant defense ideas for a
classroom environment, in particular plant resistance to plant pathogens, review Goetsch et al. (2002) and see fig.8.Plant-Herbivore Options
A good substitute caterpillar to purchase in place of the tomato fruit worm is the tobacco hornworm (Manducca sexta) which feeds on
tomato. Tobacco hornworm can be purchased from the suppliers listed. Also, one important to note that if you choose a different species
of caterpillar, you must determine if it feeds on tomatoes (or choose a different host plant). Another option for obtaining caterpillars is to
grow tomatoes in a local outdoor garden. Do not spray these tomato plants with pesticides, if your garden is in an insect friendly environment
you should get a variety of caterpillar pests. A beautiful and infamous caterpillar that can be readily reared by this method is the tomato
hornworm (Manduca quinquemaculata). This is a large green caterpillar with "tiger stripes" and an appendage that is seemingly like a
tail (practically identical to the tobacco hornworm).
Plants other than tomato may be used for induced plant defense experiments. Induced plant defenses occur among vast array of plants.
Care needs to be used when selecting another plant because you will need to make sure that it is a variety of plant that a particular insect
will feed on. The tomato fruit worm, H. zea, will feed on a vast array of crops such as corn, beans, cotton, tobacco, peppers, etc. Bean
plants could be an interesting comparison to tomato plants. The caterpillar growth could be quite different based on the levels of plant
defenses formed after previous mechanical damage for a particular plant. Although it is generally suspected that caterpillar growth will
be reduced as a result of mechanical damage to the plant, the level of caterpillar growth does not necessarily have to be less than those
found on a non-wounded plant. In an experiment Dr. Richard Musser performed with High School Students from the University of
Arkansas Math and Science Academy (an Upward Bound Program) they determined that H. zea caterpillars grew more when feeding
on tobacco plants that were mechanically wounded and treated with caterpillar saliva than those that fed on plants that were mechanically
wounded with no saliva application. It was later determined that this suppression of induced plant defenses was caused by a caterpillar
salivary enzyme glucose oxidase (Musser et al. 2002a, 2003). Conclusion
Plants have immune-like responses that are effective against herbivores, referred to as induced plant defenses (Karban and Baldwin, 1997).
These induced plant defenses are stimulated by plant damage and are an interesting phenomenon which can be tested relatively easily in most
classroom environments, and experiment complexity can be modified and designed for all levels of research experience from beginners to
leading researchers. The above experiment description can readily be adapted for solid science fair projects and undergraduate research
projects. The induced plant defense experiments can be used to teach basic experimental design, hypothesis testing and data handling in
most levels of science instruction.

References
Goetsch E., Mathias C., Mosley S., Schull M., and Brock D.L. (2002). Induced pathogen resistance
in bean plants: A model for studying "vaccination" in the classroom. The
American Biology Teacher. 58-65.
Goodman W., Jeanne R., and Sutherland P. (2001). Teaching about behavior with the
tobacco hornworm. The American Biology Teacher. 63: 258-261.
Halpern A. (2000). Toward scientific literacy for non-science majors. The American
Biology Teacher. 62: 276-281.
Karban R. and Baldwin I.T. (1997). Induced responses to herbivory. Chicago, IL: The
University of Chicago Press.
Musser R.O., Hum-Musser S.M., Eichenseer H., Peiffer M., Ervin G., Murphy J.B. and Felton G.W. (2002a). Caterpillar saliva beats
plant defences: a new weapon emerges in the coevolutionary arms race between plants and herbivores. Nature 416: 599-600.
Musser, R.O., Musser-Hum S.M., Slaten-Bickford S.E., Felton G.W. and Gergerich,
R.C. (2002b). Evidence that ribonuclease activity present in beetle regurgitant is found to stimulate virus resistance in plants. Journal of
Chemical Ecology. RC7-RC13.
Musser R.O., Farmer E., Peiffer M. and Felton G.W. (2003). Caterpillar
Salivary Gland Ablation Technique for the Clarification of the Role of the Labial
Enzyme Glucose Oxidase. (In Press Journal of Chemical Ecology).
Useful Web Sites
Tomato Anatomy:
http://www-plb.ucdavis.edu/courses/plb105/Students/Tomato/Tomhome.html
NC State Department of Entomology:
http://www.cals.ncsu.edu/entomology/insectary/homepg.html