Download Novozyme - Steve Leach-Wastewater microbiology

Wastewater
W
t
t
Microbiology
Mi
bi l
Microscopic review and
Fil
Filaments
t
Steve Leach
Novozymes
Biologicals
Disturbing Water Facts
 10 million people die every year from dirty drinking water
 40% of the worlds population has no access to basic
sanitation (toilets & running water)
 According
d
to USEPA 17% off treated
d water is lost
l
in lleaky
k
pipes
 40% of US rivers are identified as heavily polluted
 (75% in China)
population
p
will live with
 Its estimated that half the worlds p
chronic water shortages by 2050
 Underground aquifers are being depleted at higher rates
than natural
nat al processes
p ocesses can replenish
eplenish them
 AWWA Journal May 2010 issue
Bacterial Enzymatic
y
Activity
y
small, soluble compounds
Extracellular
enzymes
Intracellular
enzymes
CO2 & H2O
Membrane-bound
Membraneenzymes
P ti
Protein
G
Grease
large molecules,
molecules
particulate or colloidal
material
•
Microbiology
Bacteria
do vast majority of organic degradation.
Generation Time: replication in PURE culture:
Bacillus sp. (BOD eating bacteria) 20-30 minutes
Nit ifi
Nitrifiers
22-48
22 48 hours
h
Methanogens 10-30 days
Protozoa
Their
are useful as bacteria predators.
number and sophistication tell us what the state of
the bacterial population is
is.
A
Aeration
ti
b
basin
i
Home of active biomass
Biomass Growth Pressures
Operator
SRT
Inert material
pH
Temperature
Toxins
Dissolved Oxygen
Competition
Substrate/ concentration
Bacteria formation
Zoogloea
g
Filamentous bacteria
Free motile rod-shaped bacteria
Floc’d bacteria
Spored bacteria
Encapsulated bacteria in India Ink Stain
Brightfield View 100X
Phase contrast View 100X
Daily Microexams (on site)
 Should be performed at least 3X per week
 Limited in scope
Find Funnyy Hungry
g y Bugs
g
 Floc
 Size, shape, density
 Filaments
Fil
 Abundance, type
 Higher life forms
 Presence, abundance, diversity
 Bulk water
 Clean,, containingg inerts or bacteria
When should a detailed microexam be performed?
 When daily microexams show signs of change.
 When treatment performance is poor or worsens.
 If foaming problems occur.
 When the plant is undergoing operational changes.
 For plant confirmation and confidence
 When everything is running smoothly*
Objective progression
10X darkfield
40X Phase contrast
10X Phase contrast
40X Phase contrast
40X Phase contrast
100X Phase contrast
w/oil immersion
(Microlife / Indicator Organisms)
Amoeba
Flagellates
Swimming ciliates
Crawling ciliates
Stalked ciliates
Suctoria
Rotifers
Nematodes
A change in the microlife can
indicate a change in the
wastestream and/or plant
conditions.
P t
Protozoa
Facts
F t
 Single-celled organisms
 Feed heterotrophically
heterotrophically*
 Various forms of motility
 Eukaryotes
 Approx. 65,000 currently
identified*
g or in colonies
 Occur single
 Some are parasitic
 Oldest identified- over 500
million years (Cambrian)
(Cambrian)*
 Found almost everywhere
P t
Protozoa
Facts
F t cont.;
t
 Reproduce asexually
asexually*
 Some species are the cause
of human diseases
 Many
M
species
i form
f
cysts
t
 International Society of
Protistologists
(Protozoologists)
Single Cell- Amazing!
 Motility
 Reproduction
 Respiration
 Feeding
 Defense(encystment)
 Adaption
Protozoa
Amoebas
Flagellates
Free-swimming
F
i
i
ciliates
ili t
Crawling ciliates
Carnivore ciliates
Stalked
Stalked ciliates
Suctorians
M t
Metazoa
F t
Facts
(regards to wastewater)
 Multicellular
 Slower growing
 Worms, rotifers, gastrotrich, etc…
 Evolved
l dd
during the
h Precambrian
b
age ((545 million
ll
years ago)*
)*
 Worms and higher have approx. 55 specialized cells
 Typically larger than protozoa
 Sexual and asexual reproduction
 Heterotrophic
 All are motile*
Microexam Sample
Fil
Filament
t Facts
F t
 Filaments are chains of individual cells
 Filaments aid in Floc formation
 Filaments remove BOD
 Filaments grow in adverse conditions
 Filaments have no Boundaries
 Filaments grow in all types of WWT systems
 Filaments can create poor settling
 Filaments cost “Money”
Fil
Filament
t environment
i
t
 Low Dissolved Oxygen
yg
(D.O.)
(
)
 Low Nutrients (N and P)
 Low Food to Mass ratio ((F/M)
/ ) or High
g F/M
/
 Septicity/Organic acids
 Readily degradable substrate
 Manual on the Causes and Control of Activated Sludge Bulking, Foaming, and Other Solids
Separation Problems 3rd edition Jenkins, Richard, Daigger Lewis Publishers
Filaments and associated causes
Manual on the Causes and Control of Activated Sludge Bulking, Foaming, and Other Solids
Separation Problems 3rd edition Jenkins, Richard, Daigger Lewis Publishers
 Low Dissolved oxygen
 Low F/M
 Septicity
 Oil and Grease
 Sphaerotilus natans, Type 1701,
Haliscomenobacter hydrosis
 Type 0041, Type 0675, Type 1851,
Type 0803
 Type 021N, Thiothrix I&II, Type
0914,, Type
yp 0411,, Type
yp 0961,, Type
yp
0581, Type 0092, Nostocoida limicola
I, II, and III
 Nocardia spp., Microthrix parvicella,
Type 1863
 Nutrient Deficiency
Type 021N
021N, Thi
Thiothrix
th i I and
d II
II, S.
S
 T
natans, N. limicola III, H. hydrosis
 Low pH
 Fungi
Morphological Features
 Attached Growth:
False Branching:
 True Branching:
 Cell Shape:
Square
q
Spherical
Disc-shaped
Disc
shaped
Rectangular
Rod
Morphological Features
Constrictions:
 Crosswalls:
Sheath:
 Shape:
Straight
Curved
Coiled
N
Nocardia
di
Oil and Grease
Gram stained slide
1000X/ Bright field w/oil
Nostocoida Limicola sp.
Nutrient deficiency
Organic acids
F t
Factors
affecting
ff ti
“Chlorination”
“Chl i ti ”
 Chlorine, Peroxide, or Chlorine Dioxide
 Handling and cost influence this decision
 Exposure to Filaments ( RAS vs. Basin)
 Where is the best possible application point
 Frequency of Exposure
 A minimum of 1X per day (2-3X preferred)
 Concentration based on filament type
 Certain filaments require higher concentrations and longer exposure time
 Residual organic material
 Will the BOD/COD be oxidized by available chlorine
 Closely monitor microscopically
 Look
k for
f cell
ll changes
h
and
d possible
bl d
damage
Ch
Chemical
i l facility
f ilit Chlorination
Chl i ti
1,299
,
m3//day
y
Activated sludge (pure
oxygen)
yg )
Thiothrix II
N. limicola II
N
Type 1851
Type
T
0041
Before Chlorination
Last day of Chlorination
Last day of Chlorination
B f
Before/
/ Aft
After
Dense floc
Too many solids
Weak floc and filaments
Problem #1
Refinery
y
ASU
Solids
S lid fl
floating
ti
on
surface of secondary
Eff.
Eff TSS violation
i l i
No lab testing
available
1000X Phase
Solution
Problem
Denitrification
Increase wasting
( d
(reduce
totall
inventory)
Monitor ORP in 2nd
S
Sweet
t crude
d
Encapsulated floc
S
Sour
crude
d
Low phosphate
O
Opportunity
t it crude
d
Hyphomicrobium
Problem #2
Chemical
ASU- Pure oxygen
Scum
S
on surface
f
off
secondary
Eff.
Eff TSS violation
i l i
No lab testing
available
1000X Phase
monocolonies
Solution
Problem
Extreme Low F/M
Institute Step-feed
Inf. COD monitoring
Purchase
Purchase microscope
100X Phase
Problem #3
Paper
p Mill
ASB
Loss
L
off COD
reduction
No
N llab
b testing
i
available
400X Phase
Solution
Problem
Enzyme addition for
Pitch control
BICHEM
BICHEM 1005
addition to
compensate for
increased COD
400X Phase
High ferric addition
Unknown dump
Chlorination of filaments
Sulphide presence
Nocardia
Inter-floc bridging