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! So Get to know your neighborsthe sludge bugs!

MicroMicro-managing.the sludge! (they wont mind!)

 Observe

on a routine basis. Need to understand growth patterns. a week is good.

 Once  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. factors: availability of food source, competition for food, etc.

 Biological

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, phytoflagellates dominatedominate-take in food thru cell walls. Bacteria reproduce fast and quickly outoutcompete 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 ViewingNot 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  Put

Well.

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

Floc

Dirt

Is it Floc?

Dirt: minerals

Appearance of Floc
   

Look at under low power. Is floc wellwellformed? Is space between floc clear? Is it normal for your plant?

Appearance of Floc


Look at under high magnification. Filament statuslots, not many. Well formed floc?

Appearance of Floc

Older sludges may become more dirtdirtlike. 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.

FilamentsFilaments-Extended from Floc



Most common is Sphaerotilus, a Sphaerotilus, filamentous bacteria. Filaments branching. Low oxygen. Can grow in high oxygen, if thick floc.

  

FilamentsFilaments-Sphaerotilus sewage fungus



Grossly polluted water ways can have extensive growths of Sphaerotilus. Sphaerotilus. It is bacteria, not fungus. Great indicator organism.
Dark field illumination, stereoscope. From a creek contaminated with soft drink waste.

FilamentsFilaments-Extended from Floc

Pin Point FlocFlocundesirable, poor settling.

FilamentsFilaments-Extended from Floc

 

Sludge BBs Extended aerationwinter, reduced aeration, resulted in slow roll. Created BBs with filaments. Anaerobic in center of floc. Sunk like a rock.

FilamentsFilaments-Free from Floc



Sulphur bacteria. Can see sulphur granules in the filaments. Serious oxygen problems!

FilamentsFilaments-Free from Floc

   

Nocardia, a fungus. Free from floc. Short branches. Thrives in foam from overover-aged sludge which is the brown, greasy type. Greasiness is the polysaccharide buildup.

FilamentsFilaments-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 sludgesludgenitrous acid can form. Low N and P.

Fungal finger-like growth from a grease ball.

Other Filamentous-looking Filamentousmicrobes



Cocci chain bacteria. Lots of soluble food. Low D.O. Serious operational problems!

NextNext-Look at the Sludge Bugsbut firstis it a Sludge Bug?!



If it is alive, it probably will move! Air bubbles, cloth fibers, dirt, etc. often deceive the beginner.

Air Bubble

Is it a Sludge Bug?

Human hair-large strands hairLintLint-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 freefreeliving. 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 wormshairs. Segmented. Indicator of older sludges. Eat chunks of floc. Can be seen in supernate with the naked eye.

   

Aquatic Earthworms
   

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

Aelosoma

Aulophorus

Water Bears
    

Tartigrades MultiMulti-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 semisemi-aquatic. Generally, found more often in older sludges.

Gastrotrichs


Ciliated, multi-celled multianimal. 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


MultiMulti-celled animal. Important process control indicators. Many species-aquatic speciesand semi-aquatic, semifrom the Arctic to Antarctic! Few males known. Two major groups.

RotifersRotifers-two groups


Philodina

Bdelliod: inchworminchworm-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 freefreeswimming bacteria.

Rotifers


The inchworminchwormtype 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 Cephalodella

Common Protozoa
Flagellates Amoeba Free Swimmer Crawlers Stalked Ciliates

ProtozoaProtozoa-Flagellates  Phytomastigophora (wigglies!)

 

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.


take in food through their cell membrane. Only common when lots of food and few bacteria. Zoomastigophora are animalanimal-like in that they ingest their food, bacteria. Which type you have? Really doesnt matter to identify.

 

ProtozoaProtozoa-Flagellates
Peranema-largest, not Peranema-

unusual to have in sludge.

 

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

ProtozoaProtozoa-Flagellates


Dinobryon, Dinobryon, large, colonial flagellate. Can swim freely. Seen occasionally.

Other little wigglies



spirillium bacteria Quite visible and mobile. Lots indicate poor conditionsconditions-low oxygen.

ProtozoaProtozoa-Amoeba
Will always have some.  Lots more at startup and recovery.


ProtozoaProtozoaAmoeba


Move by pushing out cell wall. Feed by engulfing foodfoodbacteria, algae with cell wall. Tiny amoebaamoebaVahlkamfiaVahlkamfia-if lots, poor conditions.

Mayorella

Vahlkamfia

ProtozoaProtozoaAmoeba


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

CiliatesCiliates-Free Swimmer


Fine hair-like cilia for hairswimming and food gathering. Can swim freely thru water. Primarily feed on freefreeswimming bacteria. Some are predaceous. High energy needs.

CiliatesCiliates-Free Swimmers


Paramecium Rare in activated sludge. Common in trickling filters. Negatively geotactic! (they wont sink!)

CiliatesCiliates-Free Swimmers

  

Cyclidium Feed on bacteria. Tends to be more common in poorer conditions.

CiliatesCiliates-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)

CiliatesCiliates-Free Swimmers
Bacteria eater..note cytostome. Also eat diatoms and cyanobacteria.

Not uncommon even in well run plants.

Chilodenella

scrubbrush

Mouth, (Cytostome)

CiliatesCiliates-Free Swimmers


Chilodenella

ConjugationConjugation-type of reproduction

CiliatesCiliates-Free Swimmers


Trachelophyllum billy club, eat bacteria.

Trachelophyllum-mouth on very end

Not uncommon even in well run plants. Very flexible bodies.

CiliatesCiliates-Free Swimmers
 Litonotus, Litonotus,

predaceous

Litonotus-mouth on side

CiliatesCiliates-Free Swimmers


Amphileptus Note food vacuoles. Carnivorous on stalked ciliates.

(Bick)

CiliatesCiliates-Free Swimmers


Colpidium, kidney shaped. Feed on bacteria and small flagellates. (Bick) If common, likely indicating poorer conditionsespecially low oxygen.

CiliatesCiliates-Crawlers


Have thick cilia called cirri. Crawl over substances. Not so good at swimming freely. Indicators of stable sludge, Crawlers and free-swimmers were lumped in as one healthy group at the time of this chartlater separated. conditions.

CiliatesCiliates-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 freebacteria. Lower energy needs than freefree-swimmer ciliates.

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


Side view.

Suctorian having a tasty Euplotes for lunch.

CiliatesCiliatesCrawlers


Aspidica feed on bacteria. Often the most common along with the stalked ciliate, Vorticella, in a Vorticella, healthy activated sludge. ruffles potato chip

CiliatesCiliates-Crawlers


Aspidisca Reproduction via cell division.

CiliatesCiliates-Crawlers


Oxytrichs feed on algae, flagellates, and ciliates.

(Bick)

CiliatesCiliates-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.

CiliatesCiliates-Stalked Ciliates


Two major groupscolonial and single. groups Three main groups of colonial stalked ciliates in activated sludge: Epistylis, Opercularia, and Carchesium. One main group of single stalked ciliates ciliates Vorticella.

CiliatesCiliates-Stalked Ciliates

CiliatesCiliates-Stalked Ciliates
  

Epistylis Large colonies. Stalks are nonnoncontractile. Heads are like Vorticella.

CiliatesCiliates-Stalked Ciliates


Opercularia Colonial stalked ciliate. More common at start up and recovery.

CiliatesCiliates-Stalked Ciliates


Opercularia Note cap on head: operculate.

CiliatesCiliates-Stalked Ciliates


Carchesium Contractile stalk, note myoneme within stalk which provides this action. Heads are like Epistylis and Vorticella. Good sludge conditions.

CiliatesCiliates-Stalked Ciliates


Vorticella-stalk can Vorticellacontract due to myoneme. Variety of species in activated sludge. Note large nucleus. Note dots on stalk-this stalkparticular 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 freefreeswimming telotroch stage to help it get out of Dodge! Not indicative so much of toxic conditions. Usually oxygen-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.

CiliatesCiliates-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

CiliatesCiliates-Stalked Ciliates


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

Look like stalked ciliates-but ciliatesarent!



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.
Numerous suctorians, using an Epistylis colony as the substrate!

Have tentacles instead of cilia.

Mostly feed on ciliates.

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