Activated Sludge Troubleshooting Chart Unit 8 Appleton Court, Calder Business Park,

Wakefield, WF2 7AR

Tel: + 44 (0)1924 242255
Fax: + 44 (0)1924 257455
E-mail: enquiries@aquaenviro.co.uk

Independent specialists in wastewater treatment

www.aqua-enviro.net

The Microscope Examining Activated Sludge

A clean, well adjusted microscope is essential for identification of sludge microfauna. When using the microscope remember to: 1) On receipt of sample, spread a drop evenly over each of two microscope slides and allow them to air dry, whilst undertaking steps 2 and 3.
SSVI<90 SSVI>120 No SSVI ensure the condenser is setup correctly 2) Spread a drop of sample over a third slide, cover with a cover slip and gently press it down.
keep the lenses clean (always use lens paper - ethanol is good for cleaning very dirty lenses) 3) Examine under high power and phase contrast, and note:
never scratch the lens on the slide, watch from the side when making large adjustments the size and shape of the flocs

SSVI
watch from the side when changing from 10X to 40X objectives. presence of protozoa and an estimate of the predominant types
Then at a low power note:
presence of filamentous bacteria and an estimate of their abundance (on a scale of 0 to 4)

Protozoa Identification 4) After examining the wet mount, the air dried slides are used for filament identification. One of them is stained with the Gram stain, the

The Test
other using the Neisser stain.

Protozoa can Flagellated Free swimming Stalked Rotifers

comprise up to 5% of the
mass of a healthy activated
protozoa
Small (5-20m)
ciliated protozoa
Much larger
ciliates
Very easily
These are very
large
Filament Identification
sludge, with as many as and very fast (20-100m) and recognised (100-500m), Correct identification of the prevalent filament types requires a wet In addition to the above, the staining characteristics of the filaments
20,000 organisms/ml. moving protozoa slower swimming bell-shaped slow-moving and mount which is examined under phase contrast, an air-dried Gram stain are determined from the slides which received the Gram stain and
There are over 200 species with one or more than the protozoa, easily recognised. and an air dried Neisser stain. A limited number of characteristics are the Neisser stain, which are both examined under direct illumination
encountered and so whip-like flagellates. With attached by a They are not needed to identify filaments, and most of them can be seen with the (bright field).
wet mount at a magnification of 1,000X. These characteristics are: These characteristics are:
Symptoms identification to species level
is rarely carried out! They
flagella.
Because they
short, hair-like cilia
which aid their
stalk to the
sludge floc. Each
actually protozoa,
but metazoa.
Branching (present or absent, if present true or false)
Filament shape (straight, smooth-curve, bent, chains, coiled,
Gram +ve (organisms stain a blue colour)
Gram -ve (organisms stain a red colour;
are divided up into four move so fast mobility. Many of stalk can have mycelial) most filaments are gram -ve)
Poor settling sludge, many easily identifiable groups for they are difficult these protozoa can one, two or Attached growth (present or absent) Neisser -ve (whole filament stains light brown)
Sheath (present or absent) Neisser +ve (whole filament stains blue)
Good settling sludge, many filamentous bacteria, dispersed the purpose of diagnosing to see clearly. be observed numerous bells
No discernible settlement, problems with an activated crawling over the attached to each
Cross walls (present or absent) Neisser +ve granules (filament stains light brown, with dark blue
protozoa in mixed liquor, floc with few stalked protozoa Shape of cells (square, rectangular, barrel, sausage shaped) granules clearly visible)
highly dispersed floc, few or sludge plant. surface of sludge stalk.
especially stalked ciliates, and but abundant flagellates and Size (length and width of cell in m)
no protozoa - mainly flocs. Sulphur deposits
free-swimming protozoa with free-swimming protozoa. flagellates.
some rotifers. Unable to control sludge
blanket in clarifier. Vorticella Carchesium Paramecium Opercularia Thiothrix Nocardia Neisser Stain Filament Shape
(sulphur granules) (unstained) (+ve purple/blue; -ve yellow/brown)

Straight Smooth Curve

Causes
Filamentous sludge bulking or
Toxic material in influent, Coiled Mycelial
foaming. May be due to: low
organic shock loading,
Ideal operating conditions. dissolved oxygen (DO),
operating F/M too high, chronic Crosswalls Present
nutrient deficiency,
nutrient deficiency. O21N M. parvicella Gram Stain
inappropriate F/M, septicity. Aspidisca Euplotes Chilodonella (unstained) (gram stained) (+ve and -ve)

Square Rectangular

Cures Barrel
Optimise operation of
secondary sedimentation tank Undertake toxicity evaluation, Presence of Sheath
Continue this operating regime. (use the WRc nomograph) monitor influent loading,
check DO, nutrients and check nutrient levels. S.natans H. hydrosis M. parvicella
sulphide levels. Flagellated Protozoa Suctoria Rotifer Nematode (unstained) (unstained) (unstained)

Present Absent

Attached Growth

Loading Rates
Is your activated sludge plant operating well?
Absent Present
If a treatment plant receives adequate organic material for
the biomass present in the reactor (i.e. it has the correct F/M
ratio), with enough oxygen, and no toxic substances present,
Yara Industrial offers a range then a healthy sludge will develop.
of products and dosing
controllers for biological
If the F/M ratio is too high, or too low, then the result will be
sub-optimal effluent treatment. This is illustrated below.
Regular monitoring of protozoa Naming filaments
effluent treatment. The SciSol Neisser stain
Changes in protozoal populations can indicate a wide range of operating problems Free-swimming Filaments are identified and named using Filament type Gram Shape & size Cross
Sheath
and Vitamax ranges offer a Organic Loading Predominant Protozoa 100 -
ciliates a dichotomous key, and those produced
stain Filament Granule (m) walls
such as: toxicity, shock loading, inadequate nutrients and oxygen deficit. However 90 -
source of balanced nutrients and Rate (F/M) to interpret these it is important to have baseline conditions regarding your own 80 -
Stalked by Eikelboom or Jenkins are commonly Nocardia + - + Branched 1.0 x 1.0 - 2.0 - -
ciliates
trace metals; Nutriox will plant. The figure opposite is typical of an activated sludge plant - construct one 70 - employed. However in the UK a limited
Too Low Stalked ciliates and rotifers, with some nematode for your own plant as follows: 60 - Flagellated range of filament types are routinely
prevent the odour and treatability 1) Dilute the sample 1:1 with tap water and prepare a wet mount. A Lund cell protozoa encountered, and with experience these
M. parvicella + - + Irregular coils 0.6 x 100 -400 - -
worms present. 50 - Rotifers
problems caused by hydrogen allows sufficient depth of liquid for protozoal mobility, and can be calibrated to 40 - are quickly recognised. They are:
obtain a quantitative assessment of numbers. Nocardia, S. natans - - - Rounded rods 1.4 x 2.0 + +
sulphide. Ideal A large number of protozoa and a wide range of
30 -
Type 021N,
2) Count five fields of view in a vertical plane. 20 -
species. Dominated by free-swimming and stalked 3) Count protozoa in the order: flagellates, free-swimming protozoa, rotifers, stalked Type 0041, H. hydrosis - - - Straight and rigid 0.5 x 20 - 100 - +
10 -
Our customers are supported by ciliates. ciliates. 0 -| | | | | | | | | | |
Microthrix parvicella,
4) Record each group as numbers/litre mixed liquor and also note the plant sludge Sphaerotilus natans and 021N - - -, + Barrel to ovoid 1.0 - 2.0 x 1.5 - 2.0 + -
a network of field-based age, loading rate and effluent BOD. Plot them graphically and display the results.
0 1 2 4 6 8 10 16 18 20 24
Haliscomenobacter hydrosis.
Too High Large number of flagellates and small, Sludge age (d)
technical service staff. 5) Carry out the test at least three times each week (it takes only 20 minutes from Toxicity increasing Their key characteristics are summarised 0041 + - -, + Straight, smooth curve or bents + +
free-swimming ciliates. receipt of sample). Loading Rate increasing opposite: 1.4 x 1.5 - 2.0

This chart was prepared by Dr Nigel Horan and the photomicrographs supplied by Dr Louise Hornsby (School of Civil Engineering, University of Leeds).

Yara Industrial, Immingham, N. E. Lincolnshire, DN40 2NS

Tel: 01469 554711 Email: yarauk.info@yara.com

www.yara.co.uk