Session 3 - sMApp & Filamentous Organisms Flashcards
Beneficial Fungi (Deutero/Asco/Basidiomycetes)
Color?
Diameter?
Septa?
Clumps/Knots?
Oxalate Crystals?
Clean Brakes?
Color: usually different from background color (asco/basidio), but can also be colorless (deutero/asco)
Diameter: 2µm – 3.5µm (deutero) / 2 – 3.5 + (asco) / 3 – 5+ µm (basidio), most are ≥ 3µm, uniform diameter throughout each branch of hyphae
Septa: yes, can be non-uniform (deutero), but mostly uniform (asco/basidio), basidio form clamp connections
Clumps/Knots: nope
Oxalate Crystals: best examples form visible crystals
Clean Breaks: most of the time
Disease-causing fungi (Ooomycetes)
Color?
Diameter?
Septa?
Clumps/Knots?
Oxalate Crystals?
Clean Brakes?
Color: Clear, colorless, often with bubbly cytoplasm
Diameter: 1.5 – 2.5 (+) µm, not uniform throughout branches (bumpy)
Septa: no, or if so then not uniform, no clamp connections
Clumps/Knots: likely (rapid growth around food source)
Oxalate crytsals: nope
Clean Breaks: Uneven brakes can occur
Actinobacteria
Color?
Diameter?
Septa?
Clumps/Knots?
Oxalate Crystals?
Clean Brakes?
Color: clear (but usually appears as a visible black “pencil” line)
Diameter: 1 – 1.5µm, uniform throughout whole structure
Septa: nope
Clumps/Knots: possible
Oxalate crytsals: nope
Clean Brakes: most of the time
What are the criteria for recognizing actinobacteria?
What is the general shape?
What is the diameter?
- filamentous organisms that seem likes pencil line, usually a bit curved (true?)
- diameter of ca. 1μm (use smallest coccus next to it as a refernce)
- do they branch?
How do you determine length of filamentous organisms?
How do you determine the diameter?
What diameter do you record for it on sMApp?
- divide FoV horizintally in 10 units and gauge how long it is
- use smalles round coccus bacterium (in focus!) and gauge how many times it would fit into the diameter
- if the diameter is e.g. varying between 1.5- 2.5 μm, then record 2 μm
What are the criteria for recognizing oomycetes?
What is the general shape?
What is the diameter?
- filamentous, but you can see the sides of the filament (wtf does that mean?), can be uneven diameter and/or filled with bubbly cytoplasm, usually colorless and often forms clumpy structures
- 1.5- 2.5 μm, but can sometimes be wider
What are the criteria for recognizing fungi?
What is the general shape?
What is the diameter?
- filamentous, hyphae uniform diameter, septate likely, clean 90°ends brakes, not ragged, color different from background
- color can be green, purple, tan, from light brown to almost black
- diameter > 2.5 μm
Soil analysis - step by step
- open sMApp
- enter do first information
nematode scan: (10x / 100x total) scan whole cover slip, line by line including outside of the edges
- once you find one dial in 40x and ID functional group
- if necessary flame them to slow them down
main assessment: (40x / 400x total)
- use swimming lane approach, as this gives more option for adjusting to uneven distribution of sample or bubbles obstructing view
- dilute if necessary (no space between particles or view obscured)
- in each of the five lanes you select 5 random field of views along the line (without looking through the eyepiece)
- measure filamentous organims: actino, fungi, oomycetes
- count protozoa
bacterial count: (40x / 400x total)
- prepare right dilution so you can count roughly 30 bacteria in your preferred division of the FoV
- wait a few minutes for bacteria to settle
- count bacterias in 5 readings in the center of each reading area (5 face die apporach)
- only count round shaped organims, no jagged edged mineral particles, each coccus represents a bacterium (even in strepto- and staphyllococci)
Mimimum levels for BioComplete compost:
Bacterial Biomass: 135 μg/g compost
Fungal biomass: 135 μg/g compost
F:B ratio: Equal to or greater than 0.3:1
Protozoa: 10,000/g compost
Beneficial Nematodes: 100/g compost
Ciliates: must be less than 5/drop at 1:5 dilution
Minimum levels for BioComplete Liquids:
Bacterial Biomass: 135 μg/ml liquid
Fungal biomass: 135 μg/ml liquid
F:B ratio: Equal to or greater than 0.3:1(for tea, can be lower for extract)
Protozoa: 10,000/ml liquid
Beneficial Nematodes: 100/ml liquid
Ciliates: must be less than 5/drop at 1:5 dilution
How many readings/FoVs?
5 FoVs per reading
5 readings
if standard deviation ist higher than 70% it might be necessary to do more FoVs/reading
Standard Deviation
<20%: ideal, very exact results
<50%: good
up to 70%: acceptable
if your SDV is too high either do more FoVs, a new drop or the number of orgaisms is just too low to be statistically significant
If the concentration of particulates at one or both sides of the coverslip is really high, what should you do?
Use swimming lane approach and skip that part as it is not representative
