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and keep its meconium. |
Monarchs and their meconium |
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viewing through a microscope without *stain |
through a microscope |
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unstained slide resembles green jelly beans |
next to a butterfly scale |
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Buffalo Black Stain (c) Amanda Lawrence |
Buffalo Black Stain (c) Amanda Lawrence |
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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 |