The Adventures of

Sludge Bugs
Betsy Betros
Johnson County Environmental
Department
Kansas Water Environment
Association
Annual Meeting
April 3, 2007

Sludge bugs is a fun way to refer to the

microorganisms which inhabit wastewater
treatment systems.

Sludge bugs include tiny single celled

organisms such as bacteria too small to be
seen individually, to big aquatic earthworms,
visible to the naked eye.

Microbe identification
primarily for activated
sludge plants.

WWTPs are designed

to provide Utopia for
sludge bugs.

Keeping the Bugs Happy: Utopia

Happy
Bugs

Unhappy
Bugs

All they ask of us is to provide them plenty of

oxygen and food and some extra nutrition if the
wastewater is a little shy on some.

And in turn they eat and eat and eat and begat
and begat and begat lots more sludge bugs!

Nice to have some micro

critters that will eat our sh..!
Keeping the Bugs Happy
isnt always easy!

Micro-managing.the sludge!
(they wont mind!)
Observe on a routine basis.

Need to

understand growth patterns.

Once a week is good.
If problemsmay need to do more

often.

Microbes, as with all living things,

will live where they can live.
Every species has

tolerance limits.

Physical factors:

availability of nutrients,
temperature, dissolved oxygen level,
toxicity of substances, pH, etc.

Biological factors:

availability of food
source, competition for food, etc.

Its all about ecology: interactions of

organisms with each other and their
environment to provide a sustainable
biological community.

From: Microbiology for Sanitary Engineers, Ross McKinney

Bacteria are Key to the Efficiency

of
Wastewater
Treatment
Bacteria are not easily identifiable just looking at
through a microscope. Use other microbes as indicators.

Yet, it is the bacteria which are chowing* down on all

that tasty organic matter and stabilizing it by turning it
into more bacteriawhich can form floc and be removed
from the system through settling.

McKinney and Horwood discovered in the early 1950s

that many bacteria are capable of floc formation.
* Bacteria do not actually chow down on foodthey do not have mouths. A molecule of food lines up with a
reactive site on the body, and with enzymes, a chunk of the molecule is taken into the body at a time, until the
remaining portion of the molecule is no longer digestible.

When lots of soluble food

and
limited
microbial
life:
Bacteria,
Bacteria,
phytoflagellates
dominate-take in
food thru cell walls.

Bacteria reproduce
fast and quickly outcompete the
flagellates for the
food.

The phytoflagellates
die off.
Must have bacterial growth (prey)
before the protozoa (predators startup)

Microscopes

Compound Microscope vs. Stereoscope.

Compound: high magnification, light goes

through sample and image appears flat.

Stereoscope: low magnification, usually

for viewing opaque objects, using
reflected light, image is 3 dimensional
because there are two complete sets of
optics for each eye piece.

Stereo Viewing-3D
Not needed for process
control, but get a whole
different view of
microbes in 3D.

Can use transmitted

light or reflected light.

Important Features of Microscope

Use a microscope with

two eyepieces.

Mechanical stage.

3 objectives: 5X 10 X
40X or similar range.

Oil immersion difficult

to use with wet
mounts.

Phase contrast
versus flat field

Phase contrast is not

necessary for much
routine work.

Butit can be very nice, since

can see body parts much easier.

Dark Field Microscopy

Using the Microscope

Collect a sample of

MLSS.

Mix Well.
Put 1 to 2 drops on

the microscope
slide.

Get Used to Looking thru

Scope

Dont worry about

names at first.

Draw simple sketches,

make up a name.

Look for the common ones first.

Develop a way to record

your observations.

Actual counts have limited use.

Maybe use a cross between quantitative

and qualitative.

e.g. rare, occasional, common.

Variety of types is more important.

Look at Whole Slide First

-under low power

Is anything moving?

Are there any filaments?

Space between floc clear?

What are larger animals?

What is the appearance of the sludge: granular, flaky,

normal, dark, light?

Is it Floc?

Floc is made up of
living and dead
bacteria which gives
it an organic look.

Floc

Dirt is made up of
minerals which gives
it an inorganic
look.
Dirt

Is it Floc?

Dirt: minerals

Appearance of Floc

Look at under low

power.
Is floc wellformed?
Is space between
floc clear?
Is it normal for
your plant?

Appearance of Floc

Look at under high

magnification.

Filament status
lots, not many.

Well formed floc?

Appearance of Floc

Older sludges may

become more dirtlike.

Darker, denser.

Appearance of Floc

Type of aeration affects floc

size.

Inadequate shearing, develops

thicker floc which can
promote filamentous bacterial
growth which starts growing
in the anaerobic insides of the
thicker floc.

Appearance of Floc

Heavy polymer use

can yield odd
looking sludge that
still settles okay.

Next Look at Filaments

Not necessary as a
backbone.

Most sludge will

have some type of
filamentous growth.

Usually bacterial.

Filaments woven through

floccules

Most sludges will tend

to have these types, but
not often a problem.

Poor shearing of sludge

by aeration system, can
lead to extensive
growth.

Slower settling sludge.

Filaments-Extended from Floc

Most common is
Sphaerotilus, a
filamentous bacteria.

Filaments branching.

Low oxygen.

Can grow in high oxygen,

if thick floc.

Filaments-Sphaerotilus sewage
fungus

Grossly polluted
water ways can have
extensive growths of
Sphaerotilus.

It is bacteria, not
fungus.

Great indicator
organism.

Dark field illumination, stereoscope.

From a creek contaminated with
soft drink waste.

Filaments-Extended from Floc

Pin Point Flocundesirable,

poor settling.

Filaments-Extended from Floc

Sludge BBs

Extended aeration
winter, reduced aeration,
resulted in slow roll.

Created BBs with

filaments. Anaerobic in
center of floc.

Sunk like a rock.

Filaments-Free from Floc

Sulphur bacteria.

Can see sulphur

granules in the
filaments.

Serious oxygen
problems!

Filaments-Free from Floc

Nocardia, a fungus.

Free from floc.

Short branches.

Thrives in foam from

over-aged sludge which
is the brown, greasy type.

Greasiness is the
polysaccharide buildup.

Filaments-Free from Floc

Dried sludge on microscope

slide.

Sometimes
easier to assess
filaments. Can
backtrack when
it starting
developing.

Dried Nocardia

Other Filaments

Fungus growth.

Low pH.

Over digested sludgenitrous acid can form.

Low N and P.

Fungal finger-like growth

from a grease ball.

Other Filamentous-looking
microbes

Cocci chain bacteria.

Lots of soluble food.

Low D.O.

Serious operational
problems!

Next-Look at the Sludge

Bugsbut firstis it a
Sludge Bug?!

If it is alive, it
probably will move!

Air Bubble

Air bubbles, cloth

fibers, dirt, etc. often
deceive the beginner.

Is it a Sludge Bug?

Human hair-large strands

Lint-small strands.

Is it a Sludge Bug?

Polyester Fibers

Is it a Sludge Bug?

Toilet paper

Is it a Sludge Bug?

Sometimes, just wont know what it is!

Major Groups of Sludge

Bugs:
Protozoa,
Worms,
Rotifers,
Water Bears,
Gastrotrichs

Protozoa and Other Microbes

Easily viewed under a

microscope.

Excellent indicator
organisms.

With practice are fairly

easily identified.

Most feed on bacteria.

Look at Bigger
Ones to Start

Why?

Well..theyre big
and easier to see!

Two Common Worms

Round Worms
Aquatic Earthworms

Round Worms

In WWTP are freeliving.

Move by whipping
body back and forth.

Common in activated
sludge, need good
D.O.

Eat chunks.

Not usually a problem.

Not segmented.

Aquatic
Earthworms

Like garden worms-stiff hairs.

Segmented.

Indicator of older sludges.

Eat chunks of floc.

Can be seen in supernate with

the naked eye.

Aquatic
Earthworms

Aelosoma-sometimes
common.
Orange globules id it.
Aulophorus-forked
tail.
The older the sludge,
the more aquatic
earthworms.
Not usually
considered a problem.

Aelosoma

Aulophorus

Water Bears

Tartigrades

Multi-celled

Fun to watch

High survival skills!

Water bears can

survive years in a
desiccated state and
can form a cyst state.

Water Bears

Females expel eggs

into their cuticle that
they shed.
Most species are
semi-aquatic.
Generally, found
more often in older
sludges.

Gastrotrichs

Ciliated, multi-celled animal.

Appear to have a neck.

Smooth, gliding type

movement, usually
associated with a substrate.

Eat bacteria, protozoa,

algae.

Only females known.

Seed Shrimps

Ostracoda

Occasionally seen,
not important in
process control
evaluation.

Rotifers

Multi-celled animal.

Important process
control indicators.

Many species-aquatic
and semi-aquatic, from
the Arctic to Antarctic!

Few males known.

Two major groups.

Philodina

Rotifers-two
groups

Bdelliod:
inchworm-like

Ploimate: usually
with a shell
Platyias

Rotifers

Feed on chunks of floc.

Wheel animals

Cilia on head, pulls food inside.

Mastax, food grinder unique to rotifers.

Rotifers

Well run plant.

Plenty of oxygen.

Mere presence doesnt mean alls well.

Number and condition

Ext. Aer. Plants tend to have highest

diversity.

Which specific type in plant, might not

be significant.

Adineta

Rotifers can feed on chunks of

flocrather than depending on freeswimming bacteria.

Rotifers

The inchwormtype is capable of

forming a
desiccated state to
survive bad times,
particularly low
oxygen.

Important indicator.

Desiccated
Philodina

Rotifers

Monostyla

One foot

Rotifer

Cephalodella2 feet

Common Protozoa
Flagellates
Amoeba
Free Swimmer
Crawlers
Stalked Ciliates

Protozoa-Flagellates
Phytomastigophora take in
(wigglies!)
food through their cell

Tiny, wiggly.

Very common at startup

and recovery.

Will always have some.

Lots indicate lots of

soluble foodthey can
compete with bacteria at
that time.

membrane. Only common

when lots of food and few
bacteria.

Zoomastigophora are
animal-like in that they
ingest their food,
bacteria.

Which type you have?

Really doesnt matter to
identify.

Protozoa-Flagellates
Peranema-largest, not

unusual to have in
sludge.

Single cell animals.

Use a flagellum to
move and to bring
food into mouth.
Flagellum

Protozoa-Flagellates

Dinobryon,
large, colonial
flagellate.

Can swim
freely.

Seen
occasionally.

Other little wigglies

spirillium bacteria

Quite visible and

mobile.

Lots indicate poor

conditions-low
oxygen.

Protozoa-Amoeba

Will always have

some.
Lots more at startup
and recovery.

ProtozoaAmoeba

Move by pushing out

cell wall.

Feed by engulfing foodbacteria, algae with cell

wall.

Tiny amoebaVahlkamfia-if lots, poor

conditions.

Mayorella

Vahlkamfia

ProtozoaAmoeba

Shelled amoeba
testate.

They actually make

shells by cementing
together tiny sand
grains!

Arcella and Euglypha

most common.

Arcella

Arcella

Euglypha

Scanning electron
photograph of Euglypha

Made of silica.
From: The Biology of Protozoa by Michael Sleigh

Ciliates-Free Swimmer

Fine hair-like cilia for

swimming and food
gathering.

Can swim freely

thru water.

Primarily feed on freeswimming bacteria.

Some are predaceous.

High energy needs.

Ciliates-Free
Swimmers

Paramecium

Rare in activated
sludge.

Common in trickling
filters.

Negatively geotactic!
(they wont sink!)

Ciliates-Free Swimmers

Cyclidium

Feed on bacteria.

Tends to be more
common in poorer
conditions.

Ciliates-Free Swimmers

Coleps
Barrel shaped.
Move, in a spinning
rolling motion, like a
football.
Predaceous on other
protozoa.
Sensitive to NH4+ and
free ammonia.
(Bick)

Ciliates-Free Swimmers
Bacteria eater..note
cytostome. Also eat diatoms
and cyanobacteria.
Not uncommon even in
well run plants.
scrubbrush

Chilodenella

Mouth,
(Cytostome)

Ciliates-Free Swimmers

Chilodenella

Conjugation-type
of reproduction

Ciliates-Free Swimmers

Trachelophyllum

billy club, eat

bacteria.

Not uncommon
even in well run
plants.

Very flexible bodies.

Trachelophyllum-mouth on very end

Ciliates-Free Swimmers
Litonotus, predaceous

Litonotus-mouth on side

Ciliates-Free Swimmers

Amphileptus

Note food vacuoles.

Carnivorous on
stalked ciliates.

(Bick)

Ciliates-Free Swimmers

Colpidium, kidney
shaped.

Feed on bacteria and

small flagellates. (Bick)

If common, likely
indicating poorer
conditionsespecially
low oxygen.

Ciliates-Crawlers

Have thick cilia

called cirri.

Crawl over
substances.

Not so good at
swimming freely.

Indicators of
stable sludge,
healthy
conditions.

Crawlers and free-swimmers were lumped in as one

group at the time of this chartlater separated.

Ciliates-Crawlers

Euplotes

This is one similar to

Aspidisca, but more
ellipsoid in shape.

Can graze on bacteria

associated with the floc
rather than free-swimming
bacteria.

Lower energy needs than

free-swimmer ciliates.

Ciliates-Crawlers

Euplotes
Predaceous on flagellates,
algae, ciliates, bacterial
colonies. (Bick)

Side view.

Suctorian having a tasty

Euplotes for lunch.

CiliatesCrawlers

Aspidica feed on
bacteria.

Often the most common

along with the stalked
ciliate, Vorticella, in a
healthy activated sludge.

ruffles potato chip

Ciliates-Crawlers

Aspidisca

Reproduction via
cell division.

Ciliates-Crawlers

Oxytrichs feed on
algae, flagellates,
and ciliates.
(Bick)

Ciliates-Stalked Ciliates

Attached to a substrate by
a stalk.

Cilia on head for

bringing food into mouth.
All are bacteria feeders.

Usually indicative of
healthy conditions in
activated sludge.

Types and condition do

vary with activated sludge
conditions.

Ciliates-Stalked Ciliates

Two major groupscolonial and single.

Three main groups of colonial stalked

ciliates in activated sludge: Epistylis,
Opercularia, and Carchesium.

One main group of single stalked ciliates

Vorticella.

Ciliates-Stalked Ciliates

Ciliates-Stalked Ciliates

Epistylis

Large colonies.

Stalks are noncontractile.

Heads are like

Vorticella.

Ciliates-Stalked Ciliates

Opercularia

Colonial stalked
ciliate.

More common at
start up and
recovery.

Ciliates-Stalked Ciliates

Opercularia

Note cap on head:

operculate.

Ciliates-Stalked
Ciliates

Carchesium

Contractile stalk, note

myoneme within stalk
which provides this
action.

Heads are like Epistylis

and Vorticella.

Good sludge conditions.

Ciliates-Stalked
Ciliates

Vorticella-stalk can
contract due to
myoneme.

Variety of species in
activated sludge.

Note large nucleus.

Note dots on stalk-this

particular species is
V. picta.

Vorticella of poor
conditions

Tiny Vorticella.

These arent
baby vorticellas!

Poor conditions.

Stalked Ciliates of poor

conditions

Most stalked ciliates

need good oxygen.

Will try to escape if

conditions get bad.

Form a girdle of
cilia.

Stalked Ciliates of poor

conditions

Body detaches from

stalk with a freeswimming telotroch
stage to help it get out of
Dodge!

Not indicative so much

of toxic conditions.
Usually oxygen-telotroch needs to time
to form.

Stalked Ciliates of poor

conditions

Vorticella reproducing
by cell division

If lotstheyre trying
to get out of Dodge,
too!

Stalked Ciliates of poor

conditions

Vorticella can also

form a cyst to
survive extended
times of poor
conditions.

Note, the stalk is

still attached.

Ciliates-Stalked Ciliates

Reproduction is
always occurring
and it is only a
concern if there is
a sudden increase.

Why it is important
to observe your
MLSS on a regular
basis.

Epistylis cell dividing

Ciliates-Stalked Ciliates

Vaginicola has
a sheath
around the
body and does
not appear to
have a stalk.

Good
conditions.

Look like stalked ciliates-but

arent!

Suctorians

Feed on other protozoa.

This one is feeding on a

vorticella telotroch.

If lots of protozoalots
of suctorians.

Suctorians
Do have a stalk that
is attached to
substrate, cannot
contract.
Have tentacles
instead of cilia.
Mostly feed on
ciliates.

Numerous
suctorians,
using an
Epistylis
colony as
the
substrate!

Summary
Get to know

your activated sludge

microbes.
Get use to using your microscope!
Observe on a regular basis.
Develop a way to record observations.

Summary

Develop a routine way to observe.

Practice using the microscope on a regular basis.
Look at known stuff first, like TP, dirt.
Look at big picture, first.
Assess the floc condition.
Assess the filaments occurrence and type.
Look at the diversity of microbes.
Record your observations.
Relate them to the operational status of your plant.

Summary

Wastewater treatment
is about creating
Utopia for the sludge
bugs!

Summary

Why do we do this?
Dont want to go

out to a creek or
river and see
scenes like this..

Summary

We want clean water

for allboth us and the
rest of the organisms
we share this planet
with!

Visit JoCo Environmental Department

On-line: jced.jocogov.org