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Nematode Extraction Methods:Root Extraction Techniques
Kavitha GovindasamyJimmy R.RichMaria L .Mendes
A Plant-Parasitic Nematode
Sampling for Nematodes
Sampling and extracting for nematodes serves two purposes:
Diagnosis a current problem Predict a future problem
Objectives of sampling and extracting nematodes
Identify the nematode Population estimation Diagnose a nematode disease Make management decisions Regulatory purposes Research and surveys
Some Common Nematode Extraction Methods
Root Samples Jar incubation and sieving Blending and sieving Soil samples Baermann funnel Modified Baermann funnel Foliar samples Punching and incubating Scissors technique
Root Extraction Techniques:Jar Method
Principle
Nematodes are extracted based on their active migration from roots
Materials required
Glass jars and caps 325 or finer mesh sieve Labels and rubber bands Dark chamber
Sample ready for processing
Wash the roots
Damp dry the roots
Weigh the damp-dried roots
Place roots in jars, add water
Place jars in tray
Store in the dark for 72 hours
Following incubation, remove from storage and add water
Collect nematodes on sieve
Rinse nematodes from sieve
Nematode samples ready for observation
Advantages and Limitations
Advantages Extracts migratory endoparasitic nematodes Few materials needed Simple procedure to follow
Limitations Not useful for ectoparasitic nematodes Lack of aeration can be a problem Sieves are needed to concentrate nematodes
Root Extraction Techniques:Blending and Sieving
Principle
Nematodes are released from roots due to maceration.
Materials required
Blender 40 and 325 or 500 mesh sieves Pan or bowl Wire mesh Filter paper Squeeze bottle Weighing balance Scissors
Wash roots well
Cut roots
Damp dry the roots
Weigh roots
Place roots into blender, cover roots with water
Blend at low speed for 10-20 seconds
Pour the roots into 325 or finer mesh sieve
Set up a Baermann tray
Place filter paper on the Baermann tray
Add water to Baermann tray
Transfer the blended roots into the filter paper
Incubate for 24 hours
Remove roots from the pan
Pour the water in the dish through a 325 or fine mesh sieve
Concentrate nematodes for observation
Advantages and Limitations
Advantages Fast method to extract nematodes from roots Foliar nematodes can also be easily extracted
Limitations Chemicals released from the roots can be injurious to nematodes Blending for long time may damage nematodes
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
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