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Nematode Extraction Methods: Baermann Funnel and Modified Baermann Techniques

Nematode Extraction Methods: Baermann Funnel and Modified Baermann Techniques. Kavitha Govindasamy Jimmy R.Rich Maria L .Mendes. A Plant-Parasitic Nematode. Sampling for Nematodes. Sampling and extracting of nematodes serves two purposes Diagnosis a current problem

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Nematode Extraction Methods: Baermann Funnel and Modified Baermann Techniques

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  1. Nematode Extraction Methods:Baermann Funnel and Modified Baermann Techniques Kavitha Govindasamy Jimmy R.Rich Maria L .Mendes

  2. A Plant-Parasitic Nematode

  3. Sampling for Nematodes Sampling and extracting of nematodes serves two purposes • Diagnosis a current problem • Predict a future problem

  4. Objectives of sampling and extraction Nematode identification Population estimation Diagnose a nematode disease Make management decisions Regulatory purposes Research and surveys

  5. Some Common Nematode Extraction Methods • Soil samples Baermann funnel Modified Baermann funnel • Root Samples Jar incubation and sieving Blending and sieving • Foliar samples Punching and incubating Scissors technique

  6. Soil Extraction Techniques:Baermann Funnel Principle • Extraction based on the active movement of the nematodes (motility dependent method)

  7. Materials required • Funnel • Funnel stand • Wire mesh • Tissue paper/ coffee filter • Rubber or plastic tube • Clamp or spring clip • Beaker or vial • Plastic drink cup cover

  8. Baermann Funnel set up Funnel Soil sample over wire mesh Clamp to hold funnel Rubber tubing Clamp Beaker

  9. Begin Baermann funnel setup

  10. Add the soil sample to the coffee filter or tissue paper

  11. Place the sample on the funnel

  12. Cover the tissue over the soil

  13. Incubate for 48 hours and collect the nematodes

  14. Collect nematodes in a beaker

  15. Advantages and Limitations Advantages • Recovery of clean, live samples • Sieves not needed • Inexpensive to construct Limitations • Not effective for inactive nematodes • Recovery percentages may be low

  16. Modified Baermann Funnel Baermann trays(Whitehead & Hemming’s modification, 1964) Principle • Extraction based on the active migration of the nematodes (motility dependent method)

  17. Materials required • Pan or tray • Wire mesh • Tissue paper, coffee filter • Squeeze bottle • 500 mesh sieve

  18. Add water to the dish

  19. Place soil on tissue paper

  20. Place tissue paper on the wire mesh

  21. Fold the tissue over soil, add water

  22. Completed technique, ready for incubation

  23. Incubate at room temperature for 3 days

  24. Following incubation, remove wire mesh from the dish

  25. Pour water into sieve

  26. Rinse nematodes from the sieve

  27. Advantages and Limitations Advantages • Recovery of clean, live samples • Inexpensive to construct • Materials readily available Limitations • Fine soil particles cloud sample • Lack of aeration can be a problem • Sieves are needed to concentrate nematodes

  28. Acknowledgments • Janete Brito, Department of Plant Industry, Gainesville, Florida • Frank Woods, Nematode Assay Lab, University of Florida, Gainesville, Florida • Joey Orajay, Entomology and Nematology Department, University of Florida, Gainesville, FL

  29. For additional information or comments, please contact • Kavitha_nem@yahoo.com • jimmyr@ufl.edu • mlmendes@ufl.edu

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