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 JoVE Biology

Protocols for Oral Infection of Lepidopteran Larvae with Baculovirus

1, 1, 1

1Department of Entomology, Iowa State University

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    Summary

    In this video, we demonstrate oral infection techniques of lepidopteran larvae with baculovirus in order to determine insecticidal efficiency.

    Date Published: 9/03/2008, Issue 19; doi: 10.3791/888

    Cite this Article

    Sparks, W., Li, H., Bonning, B. Protocols for Oral Infection of Lepidopteran Larvae with Baculovirus. J. Vis. Exp. (19), e888, doi:10.3791/888 (2008).

    Abstract

    Baculoviruses are widely used both as protein expression vectors and as insect pest control agents. This video shows how lepidopteran larvae can be infected with polyhedra by droplet feeding and diet plug-based bioassays. This accompanying Springer Protocols section provides an overview of the baculovirus lifecycle and use of baculoviruses as insecticidal agents, including discussion of the pros and cons for use of baculoviruses as insecticides, and progress made in genetic enhancement of baculoviruses for improved insecticidal efficacy.

    Protocol

    Please visit Springer Protocols to learn more about the engineering baculovirus as an insecidical agent and the oral infection techniques used in this assay.

    Disclosures

    The authors have nothing to disclose.

    Comments

    9 Comments

    thanks a lot!
    Reply

    Posted by: AnonymousOctober 15, 2009, 11:34 AM

    I would like to know where to buy larvae.
    thanks a lot
    Reply

    Posted by: AnonymousApril 23, 2011, 7:19 AM

    You can purchase H. virescens eggs from Bioserv in New Jersey. Possibly also Agripest in North Carolina.
    Reply

    Posted by: AnonymousMay 16, 2011, 12:07 PM

    hi this a great search i need the full text please and the video thanks nd pest regards doctors
    Reply

    Posted by: AnonymousAugust 9, 2011, 8:33 PM

    Dear Ms. Sparks,
    Thanks for uploading this video. I am one of the students studying about insect bioassay. I also plan to do droplet feeding and diet plus bioassay like you. But, after watching this video, I would like to ask you some questions. (1) In droplet feeding bioassay, do you mix virus sample and food dye only?? Without adding insect diet, is it possible for the insect larvae to be willing to eat? Thus, I would like to know whether I need to add insect diet or not. (²) To determine insect lethal dose, you mentioned to arrange the mortality 1- 99%. But, I have no idea how to arrange. (3) In droplet feeding bioassay, when you drop the the virus solution dyed with calorie in 60 mm petridish, how much volume do you drop in each drop? And do the drops need to be same volume? (4) When you draw two cycles, the drop lines need to be same position? (5) After letting the larvae eat the diet infected with virus, how do you know the concentration of the virus that the larvae eat? For example, the first larvae eats one drop. the other eats two drops. As standard method to determine the concentration, which one do you use? For me, it is difficult to purchase the related articles. So, if you have some papers related that one, please could you share with me. Thanks so much.
    Best regards,
    MŒ.
    Reply

    Posted by: AnonymousSeptember 19, 2011, 3:02 AM

    (1) Yes, you mix food dye with the virus solution. The larvae are thirsty as neonates, or if using later instars, you starve them overnight so that they will readily drink. No need to add insect diet. The droplet feeding only takes about 15 minutes. (²) Take a look at Robertson and Priestler's book on Insect bioassays for full details. (3) Drops do not need to be same volume. Just use a 10 or ²0 ul pipetter for this. (4) No. (5) For neonates, you do not know the concentration ingested with the exception of species for which the amount of liquid ingested has been determined so you can calculate based on this number. Neonate larvae only ingest a few nl. More mature larvae can be fed on a known amount of virus in a given droplet. This feeding is done by hand however.
    Reply

    Posted by: AnonymousDecember 27, 2011, 12:13 PM

    Thank so much, Dr. Bonning!
    Reply

    Posted by: AnonymousJanuary 26, 2012, 2:19 AM

    Dear Dr. Bonning,
    Have a great day! Dr. Bonning, I would like to site your paper, "Protocols for Oral Infection of Lepidopteran Larvae with Baculovirus" as my reference in bioassay. Even though I am looking for the free full PDF file, I can't find it anywhere. Please could you guide me to get free full paper from any place?
    Thanks so much,
    mŒ.
    Reply

    Posted by: AnonymousApril 11, 2012, 11:06 AM

    Hi MŒ,
    Scroll up from this message and you'll see a header "Files" with the "JoVE article pdf" immediately below. Click on that for the free pdf.
    Reply

    Posted by: AnonymousApril 11, 2012, 1:22 PM

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