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Nosema in Lepidoptera - Butterfly and Moth Disease
Nosema and other diseases infect butterflies. The method to check a live butterfly for the disease nosema without harming it is to check its meconium through a microscope. Alternate methods to check for nosema require destroying the insect whether it is an egg, larva (caterpillar), pupa (chrysalis), or adult.
collect butterfly monarch meconium
monarch butterfly meconium
Emerge the butterfly
and keep its meconium.
Several cups with fresh
Monarchs and their meconium
meconium for a microscope
Meconium collected for
viewing through a microscope
without *stain
Checking meconium
through a microscope
nosema through a microscope from lepidoptera
lepidoptera nosema through a microscope
This microsporida on an
unstained slide resembles
green jelly beans
Nosema and a larger OE spore
next to a butterfly scale
nosema microsporida stained with buffalo black stain (Copyright Amanda Lawrence)
nosema microsporida stained with buffalo black stain (Copyright Amanda Lawrence)
Nosema spores stained with
Buffalo Black Stain
(c) Amanda Lawrence
Nosema spores stained with
Buffalo Black Stain
(c) Amanda Lawrence


We appreciate the Insect Pathologists that have taken time to share their knowledge and teach us about nosema and Vairimorpha imperfecta. Nosema is prevalent in corn borer and other moths in the US. Nosema can be passed from adult to offspring INSIDE the egg. Bleaching eggs will not kill nosema spores. Uninfected adult butterflies must be used for egg production to produce healthy butterflies.

* Buffalo Black Stain Recipe Courtesy of Amanda Lawrence, Mississippi State University Lepidoptera Pathology Department

0.15 g Buffalo Black stain (Napthol Blue Black)
50 ml methanol (wood alcohol)
30 ml acetic acid
20 ml distilled water

Add stain to methanol, then add water and acetic acid. Filter before use to remove any un-dissolved stain particles. Stain can be stored at 5 degrees C (41 degrees F) for up to one month.

Allow slide to air dry.
Place the slide on a warmer heated to 40 degrees C (104 degrees F) and flood with stain.
Allow to stain for 5 minutes keeping the slide flooded, not allowing the stain to dry on the slide.
Pour the residual stain from the slide into a *waste container then gently rinse the slide in water.
Blot and then allow the slide to air dry.
Observe at 1000x using oil immersion. (We use 10x eyepiece and 100x lens.)

*MSU considers the methanol and acetic acid in the stain to be hazardous waste and therefore must be discarded and stored accordingly.

Articles about nosema and Vairimorpha imperfecta:
http://cat.inist.fr/?aModele=afficheN&cpsidt=17945909
http://agspace.nal.usda.gov:8080/bitstream/10113/11284/1/IND23314132.pdf
http://journals.cambridge.org/action/displayAbstract?fromPage=online&aid=24567&previous=true&jid=PAR&volumeId=119&issueId=03