Scribd
Upload
131 views

Methods of Identification of Nematodes in The Laboratory

This document outlines various methods for identifying nematodes in the laboratory, including: extracting nematodes from soil and plant tissues using techniques like Baermann funnel, sieving, and mist extraction; observing morphological features under microscopes; and using molecular data and techniques like scanning electron microscopy, antibodies, and cytogenetics. Accurate identification requires examining characteristics like body size, stylet shape, and genital papillae under different magnifications. These identification methods help determine nematode species, genera, and relationships.

Uploaded by

palak
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPT, PDF, TXT or read online on Scribd
131 views

Methods of Identification of Nematodes in The Laboratory

This document outlines various methods for identifying nematodes in the laboratory, including: extracting nematodes from soil and plant tissues using techniques like Baermann funnel, sieving, and mist extraction; observing morphological features under microscopes; and using molecular data and techniques like scanning electron microscopy, antibodies, and cytogenetics. Accurate identification requires examining characteristics like body size, stylet shape, and genital papillae under different magnifications. These identification methods help determine nematode species, genera, and relationships.

Uploaded by

palak
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPT, PDF, TXT or read online on Scribd
You are on page 1/ 18

Methods of Identification of nematodes

in the laboratory
Nematodes

• Phylum Nematoda
• Roundworms
• Microbial Feeders
• (90% plus)
• Animal Parasitic
• Human Parasitic
• Insect Parasitic
• Plant Parasitic
INTERESTINGFacts
Interesting FACTS

• Most numerous animal


• Second most
numerous species
• Size: mostly
microscopic
• Longest is 26 feet (in
Blue whale)
• Simple morphology
• No circulatory system
• No respiratory system
• No skeleton
Examples of Non-Plant Parasitic
Nematodes
• Animal Parasites
Canine heart worms,
Ascaris
• Human Parasites
pinworms,
hookworms
• Insect Parasites
mosquitoes, mole
crickets, citrus weevils
Plant-Parasitic Nematodes
• Over 6000 known species
• Present in all ecological
niches
• Attacks almost all plants
• Cause 10% losses to
crops
• Reduces ornamental
growth
• Serious turf problems
• Big problem on tree
crops
 Nematode identification requires special techniques
to extract nematodes from soil or plant tissues
 Samples be taken when the soil is moist but not wet and be made up
of 10 to 20 small samples from an area of no more than 10 acres in
size.
 use a shovel to cut through the soil profile and then take a 1- to 2-inch
slice from the edge of the opening to simulate a core. Include feeder
roots.

 for trees, shrubs and vines, sample 12 to 15


 inches deep in a zig-zag pattern under the canopy at the drip line.

 Place the mixed sample in a plastic bag.

 Label the outside of the bag with location information using a


 permanent marker.

 Keep samples cool, below 80 °F if practical, and out of direct sunlight.


 Do not put samples in the back of trucks, trunks or no insulated
floorboards of trucks or cars. Take samples to the county

 Accurate information helps the diagnosticians do a better job.


 After this, plant parasitic nematode can be extracted from soil and
plant parts.
There are following techniques used for extraction of
nematodes :

 Extraction of nematodes from soil: Baermann funnel


technique, Cobb- sieveing and decanting method and
mist extraction method
 Extraction of cyst nematodes from soil : Fenwick-can
method
 Extraction of sluggish nematodes: Centrifugal flotation
techniques
 Extraction of nematodes from roots: Baermann funnel
technique, mist extraction method and blending
 Extraction of nematodes other plant parts: include
tubers, bulb. Stem, leaves,crowns etc. Baermann
funnel technique, mist extraction
Baermann funnel technique
Cobb- sieveing and decanting method
Fenwick-can method
 These all are methods for getting
different plant parasitic nematodes
from plant and soil samples.

 Collecting nematodes in suspension.

 Nematodes are ready for counting and


identification.
High-powered microscopes to observe minute
morphological features
Morphological Identification

 Estimate the body length and diameter of body. (0.01 -0.5


mm length)
 Using the medium power (4mm), observe the stylet. clearly
such characters as the amphids, labial and cephalic papillae,
lateral field incisures and genital papillae.

 Study all the characters shown for all stages of the nematode,
including larvae, male and female.
Scanning electron
microscope (SEM)
 Radopholus similis and R. similis citrophilus, which are not differentiated
by morpho-anatomical characters studied with a light microscope, are
readily differentiated by examining the SEM photomicrographs of the
cloacal region.
 Computers are now being used for recording and analysing identification
data.
 All such records and data can now be stored in computerized filing
systems providing databases that can be searched, retrieved and sorted
within a few minutes.

 
Genetically dependent molecular data can be used with the
morpho-anatomical characters to determine natural groupings

 Phenotypes, particularly esterases, have been used in the identification


of Meloidogyne spp. (Esbenshade and Triantaphyllou, 1990).
 polyclonal (PCA) and monoclonal (MCA) antibodies have been utilized so
far only in a limited area for the detection and identification of nematode
pests.
 Immunofluorescence could provide valuable tools for the detection of
nematode diseases and identifying and classifying plant-parasitic
nematodes.
 Serological techniques using PCA and MCA have been tried and some
specific antigens from endoparasitic nematodes have been identified
and used, for example in separating Globodera pallida from G.
rostochiensis (Schots et al., 1990).
 Cytogenetic techniques to determine chromosome number have been
used successfully in the classification of higher taxa and the
identification of species and pathotypes of Meloidogyne and
Pratylenchidae.
Thanku

You might also like