NEMATODES SAMPLING IN THREE DIFFERENT TYPES OF AREAS

A Scientific Paper submitted

in Partial fulfillment of the requirements
in Invertebrates Biology Laboratory
Under Prof. Rodgee Mae Guden

Rohanisah B. Guro

April 26 May 20, 2017

 INTRODUCTION

Soil is an excellent habitat for nematodes, and 100 cc of soil may contain several

thousand of them. Because of their importance to agriculture, much more is known about plant-

parasitic nematodes than about the other kinds of nematodes which are present in soil. Most

kinds of soil nematodes do not parasitize plants, but are beneficial in the decomposition of

organic matter. These nematodes are often referred to as free-living nematodes. Juvenile or other

stages of animal and insect parasites may also be found in soil. Although some plant parasites

may live within plant roots, most nematodes inhabit the thin film of moisture around soil

particles. The rhizosphere soil around small plant roots and root hairs is a particularly rich habitat

for many kinds of nematodes.

Nematodes play an important role in essential soil processes. The direct contribution of

nematodes to nitrogen mineralization and distribution of biomass within plants has been

demonstrated in controlled experiments. In petri-dish experiments, more nitrogen is available in

the ammonium form when bacterivorous and fungivorous nematodes are present than when they

are absent (Trofymow and Coleman, 1982).

Nitrogen mineralized through microbial grazing is available subsequently to plants

(Seastedt et al., 1988; Sohlenius et al., 1988) and has been demonstrated to affect biomass

allocation in plants. In a microcosm experiment with buffalo grass (Bouteloua gracilis), Ingham

et al. (1985) demonstrated that plant shoots grow larger in soils with bacteria, fungi, and their

respective grazers than in soils with less complex soil food webs. Root biomass may also

increase in the presence of microbial-grazing nematodes.

 Unlike earthworms, nematodes are ubiquitous and certain species are frequently the last

animals to die in polluted or disturbed areas (Freckman, 1988; Samoiloff, 1987), partly because

they can survive desiccation and revive with moisture. Relative to other soil fauna, trophic or

functional groups of nematodes can be separated easily, primarily by morphological structures

associated with their various modes of feeding (Freckman, 1988; Yeates and Coleman, 1982).

The relative abundance and size of nematodes typically make sampling and extraction easier and

less costly than for other soil fauna.

 METHODOLOGY

Sampling Process

In three different areas; pristine, polluted and decomposing, adequate amount of soil
sample was collected using a small shovel and was preferably stored in plastic or zip lock bags to
avoid dehydration. The areas are within the MSU-IIT campus. The pristine soil was collected
outside the PRISM building and the polluted soil was collected just a few meters away from it.
The decomposing soil was collected behind the COE building.

Figure1. The area where the polluted soil was collected

Extraction from the Soil

Tray Extraction Method

Instead of metal tray we used a small plastic container and attached the mesh into it. Soil
was placed in the plastic container lined with two-ply paper tissue and extracted for 3 days with
enough water. The amount of water should be in contact with the tissue paper but should not go
beyond the soil sample. The figure below shows the complete apparatus with sediments in place.

Figure2. Filling the container with enough water

Figure3. Shows the trays used to extract live free-living nematodes from the sediments.
Picking Nematodes

Fishing needle

A fish needle or handling needle can be made in several ways, for example:
1. Cut with a razor blade a fine tip to a bamboo splinter (under a dissecting microscope) and
fix it in a needle holder, or:
2. Take a hair (eyebrow hair or a pig hair; the last one is sturdier but splits easy), a fine
fishbone, a nylon hair from a toothbrush or a fine metal thread. Mount this with a drop of
glyceel on a small bamboo stick (3 cm) ,which is fixed in a needle holder, or on the tip
of a dissecting needle. If necessary the tip can be cut slantwise under a dissecting
microscope. Soft material (hairs) reduces the chance of damaging nematodes, but makes
it harder to pick heavy nematodes. When working under sterile conditions, for example if
nematodes have to be transferred to agar plates (Chapter 4), a handling needle with a
metal thread is required, because it can be sterilized in a flame. A cats whisker, which is
spatulate, is suitable for picking nematode eggs.

Mounting

Nematode mounts can be made on ordinary glass slides. Glass slides are cheaper, and are used
for temporary mounts. The advantage is that the mounts can be observed from either side and
breaking is less probable as the glass slide is fixed between pieces of cardboard, thicker than the
glass. The mounts can also be piled up without touching each other.

Dissecting or Compound Microscope

Either of the two can be used to study the structure of the nematodes.
 RESULTS AND DISCUSSION

Figure1. Nematode from Pristine Environment

The proportion of bacterial-feeding and fungal-feeding nematodes is related to the

amount of bacteria and fungi in the soil. Commonly, less disturbed soils contain more predatory
nematodes, suggesting that predatory nematodes are highly sensitive to a wide range of
disturbances.
 Figure2. Nematode A from Polluted Environment

They are susceptible to soil disturbance and are often absent from disturbed, polluted, or
intensely-managed environments (Bongers, 1990, 1999). The biology of many of the taxa in cp
classes 35 is not well known; further study may reveal physiological and behavioral
characteristics that warrant some realignment of functional guilds.
 Figure3. Nematode B from Polluted Area

Nematodes are unsegmented roundworms. Not all nematodes are bad; in fact, most
species are beneficial, feeding on bacteria, fungi, or other microscopic organisms. There are even
nematodes that can be used as biological control organisms to help manage important turf insect
pests. However, there also are genera of nematodes that are pests or pathogens of animals or
plants. Nematodes that feed on plants are called plant-parasitic nematodes. Plant-parasitic
nematodes are very small, and microscopes are required to see them (Figure 2). Plant-parasitic
nematodes have a stylet or mouth-spear that is similar in structure and function to a hypodermic
needle (Figure 3a, b). The stylet is used to puncture plant cells, and then the nematode can inject
digestive juices and ingest plant fluids through it. Plant-parasitic nematodes are divided into
groups based on how they feed on plants.
Figure4. Nematode from Decomposing Area
 CONCLUSION

The results show that different types of environment drive modifications in the
composition of nematode communities based on ecological traits conferring local adaptations.
this study highlights the potential of nematodes' derived ecological indices in understanding
ecosystem processes along ecological gradients. Future accurate sampling strategies of
nematodes across more specific habitats are nevertheless required to dissect how ecosystems
types and ecological factors affect nematode-driven soil ecosystem processes along elevation
gradient.
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