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Culturing beneficial nematodes

Fungal-feeding nematodes have small, narrow stylets, or spears, in their stoma (mouth) which they use to puncture thecell walls of fungal hyphae and withdraw the cell fluid. This interaction releases plant-available nitrogen from fungal biomass.



This bacterial-feeding nematode, Elaphonema, has ornate lip structures that distinguish it from other nematodes. Bacterial-feeders release plant-available nitrogen when they consume bacteria.



Root-feeding nematodes use their stylets to puncture the thick cell wall of plant root cells and siphon off the internal contents of the plant cell. This usually causes economically significant damage to crops. The curved stylet seen inside this nematode is characteristic of the genus Trichodorus.



The Pratylenchus, or lesion nematode, has a shorter, thicker stylet in its mouth than the root feeder



Most nematodes in the soil are not plant parasites. Beneficial nematodes help control disease and cycle nutrients.



___________________________________________________________________
THE LIVING SOIL: NEMATODES

Nematodes are non-segmented worms typically 1/500 of an inch (50 µm) in diameter and 1/20 of an inch (1 mm) in length. Those few species responsible for plant diseases have received a lot of attention, but far less is known about the majority of the nematode community that plays beneficial roles in soil.

An incredible variety of nematodes function at several trophic levels of the soil food web. Some feed on the plants and algae (first trophic level); others are grazers that feed on bacteria and fungi (second trophic level); and some feed on other nematodes (higher trophic levels).

Free-living nematodes can be divided into four broad groups based on their diet. Bacterial-feeders consume bacteria. Fungal-feeders feed by puncturing the cell wall of fungi and sucking out the internal contents. Predatory nematodes eat all types of nematodes and protozoa. They eat smaller organisms whole, or attach themselves to the cuticle of larger nematodes, scraping away until the prey’s internal body parts can be extracted. Omnivores eat a variety of organisms or may have a different diet at each life stage. Root-feeders are plant parasites, and thus are not free-living in the soil.

WHAT DO NEMATODES DO?

Nutrient cycling. Like protozoa, nematodes are important in mineralizing, or releasing, nutrients in plant-available forms. When nematodes eat bacteria or fungi, ammonium (NH4+) is released because bacteria and fungi contain much more nitrogen than the nematodes require.

Grazing. At low nematode densities, feeding by nematodes stimulates the growth rate of prey populations. That is, bacterial-feeders stimulate bacterial growth, plant-feeders stimulate plant growth, and so on. At higher densities, nematodes will reduce the population of their prey. This may decrease plant productivity, may negatively impact mycorrhizal fungi, and can reduce decomposition and immobilization rates by bacteria and fungi. Predatory nematodes may regulate populations of bacterial-and fungal-feeding nematodes, thus preventing over-grazing by those groups. Nematode grazing may control the balance between bacteria and fungi, and the species composition of the microbial community.

Dispersal of microbes. Nematodes help distribute bacteria and fungi through the soil and along roots by carrying live and dormant microbes on their surfaces and in their digestive systems.

Food source. Nematodes are food for higher level predators, including predatory nematodes, soil microarthropods, and soil insects. They are also parasitized by bacteria and fungi.

Disease suppression and development. Some nematodes cause disease. Others consume disease-causing organisms, such as root-feeding nematodes, or prevent their access to roots. These may be potential biocontrol agents.

WHERE ARE NEMATODES?

Nematodes are concentrated near their prey groups. Bacterial-feeders abound near roots where bacteria congregate; fungal-feeders are near fungal biomass; root-feeders are concentrated around roots of stressed or susceptible plants. Predatory nematodes are more likely to be abundant in soils with high numbers of nematodes.

Because of their size, nematodes tend to be more common in coarser-textured soils. Nematodes move in water films in large (>1/500 inch or 50 µm) pore spaces.

Agricultural soils generally support less than 100 nematodes in each teaspoon (dry gram) of soil. Grasslands may contain 50 to 500 nematodes, and forest soils generally hold several hundred per teaspoon. 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.

NEMATODES AND SOIL QUALITY

Nematodes may be useful indicators of soil quality because of their tremendous diversity and their participation in many functions at different levels of the soil food web. Several researchers have proposed approaches to assessing the status of soil quality by counting the number of nematodes in different families or trophic groups.* In addition to their diversity, nematodes may be useful indicators because their populations are relatively stable in response to changes in moisture and temperature (in contrast to bacteria), yet nematode populations respond to land management changes in predictable ways. Because they are quite small and live in water films, changes in nematode populations reflect changes in soil microenvironments.

http://www.google.com/imgres?imgurl...0&sa=X&oi=image_result&resnum=1&ct=image&cd=1
 
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Scay Beez

Active member
Tactical Farmer: Those pics rule man, thanks for taking the time to post them up the different species!

CT GUY: It is a huge blessing to have someone in the industry aboard. When I get the cash, I'm going to buy a really nice scope. I want one that I can hook up to a computer or TV and get down to business. What magnification level does it take just to recognize that nematodes are present? I figure most folks are going to start with a culture purchased from a beneficial insect seller so identification is not necessary. Out of curiosity -> What magnification level will allow one to differentiate species?

So after a little oatmeal reading it seems I don't want to use instant or quick cooking.

Here's a simple baby oatmeal recipe I found:

Oatmeal Cereal

1/4 cup of ground oats (do NOT use instant or Quick Cook), ground in blender or food processor
3/4 cup - 1 cup water

1. Bring liquid to boil in saucepan. Add the oatmeal powder while stirring constantly.
3. Simmer for 10 minutes, whisking constantly
3. Serve warm. (*Cool down before adding nematodes*)


- sbz
 
C

CT Guy

Sbz,

The photos that were posted are excellent examples! My only caveat is that when you actually are looking in a microscope, it will look closest to that bottom photo. It becomes a little more challenging to identify. I'm guessing the bottom photo in the group was taken at 200x magnification, and the others are probably 400x. Another thing to consider is that since we're looking at live samples, they rarely hold still long enough to get a very long look at the mouth, and then the higher the magnification, the trickier it gets to keep them in your field of view.

I think I mentioned that microscope advisory already...I'd get a scope that you can see the bacteria and fungi too while you're at it. Typically, one that goes up to 400x. Mine goes to 1000x, but then it's oil immersion and you don't need it for the teas (it's more just for fun). Depending on what you want to spend, you can get a decent scope for a few hundred dollars. Mine is a $4000 scope, but then we wanted to have phase contrast, as it makes identification even easier.

I'll have to try out the baby oatmeal recipe. We've just been buying Gerber or other commercial brands.
 

sophisto

Member
CT guy, scay beez, jay kush...You guys always impress me with your knowledge and love of what you do....

Great thread and great posts...


Is it possible to overpopulate the soil with nematodes???? Seems like with compost teawe can accuire not just nematodes but all the other parts of the soil biology that are just as importan as our worm like friends..

New to all this so I could be off...
 

Kenny Lingus

Active member
Steinernema Feltiae

Steinernema Feltiae

Tactical Farmer said:
THE LIVING SOIL: NEMATODES

Nematodes are non-segmented worms typically 1/500 of an inch (50 µm) in diameter and 1/20 of an inch (1 mm) in length. Those few species responsible for plant diseases have received a lot of attention, but far less is known about the majority of the nematode community that plays beneficial roles in soil.

An incredible variety of nematodes function at several trophic levels of the soil food web. Some feed on the plants and algae (first trophic level); others are grazers that feed on bacteria and fungi (second trophic level); and some feed on other nematodes (higher trophic levels).

.....Predatory nematodes eat all types of nematodes and protozoa. They eat smaller organisms whole, or attach themselves to the cuticle of larger nematodes, scraping away until the prey’s internal body parts can be extracted.....


KOPPERT (NL) Biological Pest control producer, claim their Steinernema f. enters larvae orifices and digest them from within. Leaves corpse of for example fungus gnat larvae gray/slimy and hard to detect in the medium. They come in a substrate based on clay and will survive for 1-3weeks after receiving and are susceptible to mold. (However they are best used immediate and may reproduce in medium. (I've seen the thrive and hunt by scouting/smelling their surroundings with their slender bodies above the surface for 2 weeks after release, but then diminish. (one real dry spell and they're reduced by great numbers. koppert.nl say application may be needed twice the first two weeks and then once every sixth week.
(I get a bunch enough for spraying over 100sqm!!! So I always end up squirting around the house and trying to get my neighbors to use a little in their houseplants soil too, but now I will try to cultivate my own for reapplications. The blister pack I get is costing me like 80usd or so.)

On the other hand Predatory mites like Hypoaspiis is really easy to keep alive in a perpetual garden, but I don't find any of them being seriously effective.
 
C

CT Guy

There was a previous link that showed some photos for ID of nematodes, but it's not too bad, just look at the mouth. Also, I've found that when I inoculate a good compost with baby oatmeal, I get a huge increase in bacteria feeding nematodes after a week or so (along with large diameter fungal hyphae)
 
G

Guest

So am I correct in assuming that ew castings under the brand name Wiggle Worm contains all the good species of nematodes?
 

MrFista

Active member
Veteran
Bump - I vote this thread gets stickied.

Great link Dignan.

Learn something new every day...

The mystery of why Terra Preta strips nitrogen has now been answered to my full satisfaction.

And nematodes (among others) are responsible for re-releasing the nutrients to make them plant available. Quite the eye opener!

I don't know where I read it but aren't the plant root feeding nematodes larger than others, making a screening process possible?

Waterways (clean) have adenophorean nematodes (bacterial feeders) and so should be a good source of starting cultures.

Fascinating stuff.
 

Tilt

Member
I had a problem with fungus gnats. I ordered up some beneficial nematodes and inoculated everything houseplants,water traps for sinks even outside my place. Gnats disappeared over next few weeks. A few months later I bring home some new soil to supplement what I had. Bamm gnats are back and no money to inoculate so I mixed my old soil with the new and gnats disappear again. My conclusion is I have a healthy nematode population going and it is a good idea to recycle my soil.
 

Microbeman

The Logical Gardener
ICMag Donor
Veteran
mixing oat flour (or powdered oatmeal) about 1:20 with your compost and keeping damp and covered with a cloth for 8 to 10 days. This does work, although I am unsure whether there is a diversity of species of fungal hyphae grown. If you see white or blue fuzz growing on the surface turn it under. What we want is transparent and colored microscopic fungal hyphae. A side benefit to this procedure is that if left longer than 10 days I have seen multitudes of bacterial feeding nematodes growing. I’m not sure if this is peculiar to my compost. Try it. Compost tea is not a good medium for distributing nematodes. Better to distribute them by hand in the compost.

From my webpage; been there a few years. Surprised you guys never noticed. There really is a lot of info on there.

I've attached a few of my photos below, one of which is a close up of a bacterial feeding nematode with mouth closed. Another shows the mouth partly open. The species of bacterial feeders I have exhibit a goldfish type action with their mouth. Using compost or better yet, vermicompost is a good source of the good guys. Collecting nematodes to culture willy nilly for feeding fish is great but I would not do this for my soil. You may be multiplying root feeders. Although judging by the description in the literature provided, I would think only bacterial or organic matter feeders would be multiplied. In my best knowledge, the way it works is that adding the wet 'flour' promotes fungi which promotes bacteria (& maybe fungal feeding nematodes) which promotes bacterial feeding nematodes. I will be doing an experiment this fall/winter utilizing cheap wheat bran to see if it works as well.

I have pretty good microscopes for sale along with cameras for computer interface. I'm Tad's buddy who wrote the microscope adisory he referenced.
 

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Microbeman

The Logical Gardener
ICMag Donor
Veteran
Here's a nice breakdown that covers what the Soil Biology Primer does, but in a more digestible way:

http://www.soilsecrets.com/Soil Ecology and the Soil Food Web.pdf

This is from the guy who argues that it was he and not Elaine Ingham who first described the Soil Food Web. Note he claims it as trademark or something. [Interesting all the 1985+ citations in a 1975 paper.] Actually in reality it was a little known European, Vigdis Torsvik
 

Dignan

The Soapmaker!
Veteran
*I* first described the soil food web. I mean, Soil Food Web (TM).

I also invented the internet.
 

DARC MIND

Member
Veteran
beta
please keep us posted on ur progress
i tried this method a while back and just got a hard dry muck
best of luck and any tips or advice on ur success would be much appreciated
 

Microbeman

The Logical Gardener
ICMag Donor
Veteran
Can nematode cultures becomes 'infected' with detrimental nematodes? Can nematodes be overapplied? I have what I think is a root aphid infestation, and would like to use this culturing method to supply me with enough nematodes to re-innoculate every other day or so until they're gone. I'll be sure to doccument my progress, or lack thereof...

Collecting nematodes to culture willy nilly for feeding fish is great but I would not do this for my soil. You may be multiplying root feeders.

From post earlier in this thread.

Yes you could end up propagating the wrong species by gathering them here and there. I'd use a reliable (vermi)compost source.
 

floral

Member
Love this thread.

I think I accidentally ended up culturing nematodes the last time I was "activating" compost to encourage more fungal development before brewing tea. Life intervened and the container of compost with ground-up oatmeal etc. soon started pulsing with what I assume were nematodes.

In response to the sighting of a few gnats, I'm about to introduce some nematodes to my outdoor garden in the next couple days and am wondering if it would make sense to apply compost tea before the nematodes, after the nematodes, or both? I would guess I'd want the nematodes on arrival to just seek out any pest larvae, and then maybe add some tea to boost the potential prey species a day or two later, but maybe that's totally wrong. Any thoughts from the microbe experts?
 

floral

Member
I read here that only the "infective juvenile" stage of entomopathogenic nematodes will live outside of a host insect. Does that mean there's no point in trying to culture your own predatory nematodes in a medium, that in order to propagate them effectively you need to expose an insect and wait for the nematodes and their bacteria comrades to bust out with an "Alien"-like explosion of new "infective juvenile"-stage nematodes?
 
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floral

Member
Holy crap, never mind. Put a dead beetle in there that I found outside and looking at it with my 30x magnifier I see the beetle is covered with tiny writhing worms. I'm almost - ALMOST! - wishing to find a caterpillar or leafhopper so I can put it in there and watch it become a nematode hotel.

Still confused as to whether the cereal mush would have been sufficient to multiply the nematodes. There must be a way to reproduce them outside of bug hosts or companies wouldn't be able to sell them, right? But is my mushy oatmeal going to be an adequate substitute for their lab gear, who knows.
 

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