Identification & Control of Filamentous Bacteria
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1 Identification & Control of Filamentous Bacteria Toni Glymph, Wastewater Microbiologist ARWA-Lonoke
2 Filamentous Bacteria Sludge Bulking
3 Filamentous Bacteria Sludge Foaming
4 FILAMENTOUS BACTERIA In the activated sludge treatment system, bacteria may occur singly, or in small chains or clumps. Shifts in the activated sludge environment such as changes in ph, dissolved oxygen, nutrients etc. will often cause a change in the behavior of the bacteria.
5 FILAMENTOUS BACTERIA In stead of single cells, small chains or clumps, the bacteria will begin to form longer chains which develop into filamentous bacteria.
6 Filamentous Bacteria Round bacteria will form a chain with other round bacteria, square with square, rectangle with rectangle etc.
7 FILAMENTOUS BACTERIA (oval) (discoid) (round) Cocci (rod-shaped) Bacillus (square) (rectangle)
8 Typical Observation Filament Shape Filament Size Cell shape Cell size Cell Septa Indentations Sheath (present/absent) Branching (true/false) Epiphyte (attached growth) Motility
9 Typical Observation: Filament Shape Straight Smoothly Curved Irregular
10 Typical Observation: Filament Size Filament length Filament width
11 Typical Observation: Cell Shape
12 Typical Observation: Cell Size Length of cell Width of cell
13 Typical Observation: Cell Septa Cell septa with no indentation Cell septa with indentation Cell septa not clearly seen
14 Typical Observation: Sheath Sheath Individual cells within a sheath
15 Typical Observation: Branching True branching Fungus False branching S. natans
16 Typical Observation: Epiphyte (Attached Growth)
17 Typical Observation: Motility
18 Microscope Requirements Phase contrast Microscope 10X or 20 X objective 40 X objective 100 X (oil immersion lens)
19 Stains Gram Stain Neisser Stain
20 Gram Stain Separates bacteria into two categories Determined by the bacteria cell wall Gram (+) bacteria cell wall retains the purple color Gram ( - ) bacteria cell wall loses the purple color Gram positive bacteria are purple Gram negative bacteria are pink
21 Gram Stain Gram positive Gram negative Polysaccharide Slime Layer Cell Wall Lipopolysaccharide Slime Layer Phospholipid Lipoprotein Cell Wall Cell Membrane Cell Membrane
22 Gram Stain Gram (+) Nocardia sp. Gram (-) Type 1701
23 Gram variable Type 0041 Gram Stain
24 Neisser Stain Separates bacteria into two categories Distinguishes the bacteria which contain granules that accumulate polyphosphate Neisser (+) bluish color Neisser ( - ) brownish color
25 Neisser Stain Neisser (+) Nostocoida limicola Neisser (+) granules
26 Neisser Stain Neisser (+) Type 0092 Neisser (+) Nostocoida limicola and Neisser (-) Thiothrix
27 Slide Preparation Wet Mount Smear Dried for staining
28 Slide Preparation: Wet Mount A drop of a well mixed representative sample is place on a clean grease-free slide. A clean cover-glass is placed on top. Avoid entrapment of air as much as possible. The size of the drop is critical Use a pipette with a small opening
29 Slide Preparation: Wet Mount Drop Cover glass Slide
30 Slide Preparation: Wet Mount This allows the observer to view the microorganisms live in their environment. With a live sample, measurements can be taken and cell shape and size can be accurately determined.
31 Slide Preparation: Smear Place a drop of sample on a clean, wax-free slide and smear across the slide. Let the slide air dry. DO NOT HEAT FIX!
32 Slide Preparation: Smear Slide
33 Sample Collection For bulking sludge, collect samples from the discharge end of the aeration basin. For foaming sludge, collect a sample of the foam.
34 Filamentous Bacteria Most filamentous bacteria in Activated Sludge are associated with low F/M along with some other factor(s) that uniquely gives them an advantage over the other microorganisms. Discovering and changing what gives them the advantage will help control their dominance.
35 Filamentous Bacteria A Simpler Approach Bulking Filaments Foaming Filaments
36 Filamentous Bacteria Bulking Filaments Collect a sample of the mixed liquor from the discharge end of the aeration basin Make a smear Gram stain Neisser stain (optional) Wet Mount
37 Filamentous Bacteria A Simpler Approach Nostocoida limicola Sphaerotilus natans Haliscomenobacter Hydrossis Thiothrix I & II Microthrix parvicella Nocardia Beggiatoa Type 0041 Type 0675 Type 0092 Type 1851 Type 1701 Type 021N Type 0914 Type 0961 Type 1863
38 Filamentous Bacteria Bulking Filaments Gram Stain Gram (+) (purple) Nostocoida limicola Microthrix parvicella Gram Variable Type 0041 Type 0675 Type 1851 Gram (-) (pink) Investigate further
39 Filamentous Bacteria Bulking Filaments Gram Stain Gram (+) (purple) Nostocoida limicola Microthrix parvicella Gram Variable Type 0041 Type 0675 Type 1851 Gram (-) (pink) Investigate further
40 Bulking Filaments Nostocoida limicola
41 Bulking Filaments Nostocoida limicola
42 Bulking Filaments Nostocoida limicola This filament is associated with Low F:M Starch containing wastes (potato processing) More commonly found in industrial waste plants
43 Bulking Filaments Microthrix parvicella
44 Bulking Filaments Microthrix parvicella This filament is associated with Low F:M Animal and vegetable greases, oils and fats More common in colder temperatures Also causes foaming
45 Filamentous Bacteria Bulking Filaments Nostocoida limicola Sphaerotilus natans Haliscomenobacter Hydrossis Thiothrix I & II Microthrix parvicella Beggiatoa Type 0041 Type 0675 Type 0092 Type 1851 Type 1701 Type 021N Type 0914 Type 0961
46 Bulking Filaments Gram Stain Gram (+) (purple) Gram Variable Gram (-) (pink) Nostocoida limicola Microthrix parvicella Type 0041 Type 0675 Type 1851 Investigate further
47 Bulking Filaments Type 0041 Type 0675 Type 1851 Gram Variable
48 Gram Variable Type 0041
49 Gram Variable Type 0041 This filament is associated with Low F:M Nutrient deficient conditions
50 Gram Variable Type 0675
51 Gram Variable Type 0675 This filament is associated with Low F:M Nutrient deficient conditions
52 Gram Variable Type 1851
53 Gram Variable Type 1851
54 Gram Variable Type 1851 Bundles
55 Type 0041 Type 0675 Type 1851 Gram Variable All three are Gram Variable All three are favored under the same conditions Low F/M and Low nutrients
56 Filamentous Bacteria Bulking Filaments Nostocoida limicola Sphaerotilus natans Haliscomenobacter Hydrossis Thiothrix I & II Microthrix parvicella Beggiatoa Type 0041 Type 0675 Type 0092 Type 1851 Type 1701 Type 021N Type 0914 Type 0961
57 Bulking Filaments Gram Stain Gram (+) (purple) Gram Variable Gram (-) (pink) Nostocoida limicola Type 0041 Type 0675 Type 1851 Investigate further
58 Bulking Filaments Neisser Stain Neisser (+) (bluish) Nostocoida limicola Type 0092 Neisser (-) (brownish) All others
59 Bulking Filaments Neisser Stain Neisser (+) (bluish) Nostocoida limicola * Type 0092 Neisser (-) (brownish) All others
60 Filamentous Bacteria Bulking Filaments Nostocoida limicola Sphaerotilus natans Haliscomenobacter Hydrossis Thiothrix I & II Microthrix parvicella Beggiatoa Type 0041 Type 0675 Type 0092 Type 1851 Type 1701 Type 021N Type 0914 Type 0961
61 Neisser Positive Nostocoida Limicola
62 Neisser Positive Type 0092
63 Neisser Positive Type 0092 This filament is associated with Low F:M Excess oils and grease Colder temperatures
64 Filamentous Bacteria Bulking Filaments Nostocoida limicola Sphaerotilus natans Haliscomenobacter Hydrossis Thiothrix I & II Microthrix parvicella Beggiatoa Type 0041 Type 0675 Type 0092 Type 1851 Type 1701 Type 021N Type 0914 Type 0961
65 Bulking Filaments Neisser Stain Neisser (+) (bluish) Nostocoida limicola Type 0092 Neisser (-) (brownish) All others
66 Bulking Filaments Other Characteristics Sheath Epiphyte (Attached growth) Sulfur granules Motility
67 Bulking Filaments Sheath Sheath w/epiphyte Type 1701 Type 0041 Type 0675 Type 1851 Sheath w/o Epiphyte Sphaerotilus natans Thiothrix I & II Haliscomenobacter hydrossis
68 Bulking Filaments Sheath Sheath w/epiphyte ( Attached Growth) Type 1701 Type 0041* Type 0675* Type 1851*
69 Filamentous Bacteria Bulking Filaments Nostocoida limicola Sphaerotilus natans Haliscomenobacter Hydrossis Thiothrix I & II Microthrix parvicella Beggiatoa Type 0041 Type 0675 Type 0092 Type 1851 Type 1701 Type 021N Type 0914 Type 0961
70 Sheath w/epiphyte Type 0041
71 Sheath w/epiphyte Type 0675
72 Sheath w/epiphyte Type 1851
73 Sheath w/epiphyte Type 1701
74 Type 1701
75 Sheath w/epiphyte Type 1701 This filament is associated with low DO conditions
76 Filamentous Bacteria Bulking Filaments Nostocoida limicola Sphaerotilus natans Haliscomenobacter Hydrossis Thiothrix I & II Microthrix parvicella Beggiatoa Type 0041 Type 0675 Type 0092 Type 1851 Type 1701 Type 021N Type 0914 Type 0961
77 Sheath w/o Epiphyte Sheath w/o Epiphyte (Attached growth) Sphaerotilus natans Thiothrix I & II Haliscomenobacter hydrossis
78 Sheath w/o Epiphyte Sphaerotilus natans
79 Sheath w/o Epiphyte Sphaerotilus natans
80 Sheath w/o Epiphyte Sphaerotilus natans S. natan also exhibits false branching Associated with conditions where there is inadequate DO for the applied organic loading
81 Sheath w/o Epiphyte Thiothrix I & II Type II Type I
82 Sheath w/o Epiphyte Thiothrix I & II This filament is associated with The presence of reduced sulfur compounds (septic wastes) Wastes deficient in nitrogen The presence of organic acids
83 Sheath w/o Epiphyte H. Hydrossis
84 Sheath w/o Epiphyte H. Hydrossis This filament is associated with low DO conditions
85 Filamentous Bacteria Bulking Filaments Nostocoida limicola Sphaerotilus natans Haliscomenobacter Hydrossis Thiothrix I & II Microthrix parvicella Beggiatoa Type 0041 Type 0675 Type 0092 Type 1851 Type 1701 Type 021N Type 0914 Type 0961
86 Bulking Filaments Sulfur Granules Type 021N Thiothrix I & II* Type 0914 Beggiatoa
87 Sulfur Granules Thiothrix I
88 Sulfur Granules Thiothrix I
89 Sulfur Granules Thiothrix II
90 Bulking Filaments Sulfur Granules These filaments are all associated with The presence of reduced sulfur compounds (septic wastes) Wastes deficient in nitrogen The presence of organic acids
91 Sulfur Granules Type 021N
92 Sulfur Granules Type 021N Stacked (hockey puck) cells
93 Sulfur Granules Type 0914
94 Sulfur Granules Beggiatoa
95 Filamentous Bacteria Bulking Filaments Nostocoida limicola Sphaerotilus natans Haliscomenobacter Hydrossis Thiothrix I & II Microthrix parvicella Beggiatoa Type 0041 Type 0675 Type 0092 Type 1851 Type 1701 Type 021N Type 0914 Type 0961
96 Bulking Filaments Beggiatoa* Motility
97 Bulking Filaments Motility Beggiatoa is the only filament that is motile The presence of this filament is associated with Organic overloading Septic conditions Most often found in trickling filters and RBCs
98 Bulking Filaments Beggiatoa
99 Filamentous Bacteria Bulking Filaments Nostocoida limicola Sphaerotilus natans Haliscomenobacter Hydrossis Thiothrix I & II Microthrix parvicella Beggiatoa Type 0041 Type 0675 Type 0092 Type 1851 Type 1701 Type 021N Type 0914 Type 0961
100 Bulking Filaments Type 0961 Not Gram + Not Neisser + No sheath No attached growth No sulfur granules Not motile
101 Bulking Filaments Type 0961
102 Bulking Filaments Type 0961
103 Bulking Filaments Type 0961 Low F:M Low soluble BOD
104 Filamentous Bacteria Foaming Filaments All foaming filaments are associated with: Excess greases, oils and fats Low F:M Hydrophobic cell walls
105 Filamentous Bacteria Foaming Filaments Collect a sample of the foam Make a smear Gram stain
106 Filamentous Bacteria Foaming Filaments Type 1863 Microthrix parvicella Nocardia
107 Filamentous Bacteria Foaming Filaments Gram Stain Gram (+) (purple) Microthrix parvicella Nocardia Gram (-) (pink) Type 1863
108 Filamentous Bacteria Foaming Filaments Gram Stain Gram (+) (purple) Microthrix parvicella Nocardia Gram (-) (pink) Type 1863
109 Foaming Filaments Type 1863
110 Foaming Filaments Type 1863
111 Foaming Filaments Type 1863 Type 1863 is associated with Excess grease and fat Low F:M Decline in aeration basin ph
112 Filamentous Bacteria Foaming Filaments Type 1863 Microthrix parvicella Nocardia
113 Filamentous Bacteria Foaming Filaments Gram Stain Gram (+) (purple) Microthrix parvicella Nocardia Gram (-) (pink) Type 1863
114 Filamentous Bacteria Microthrix parvicella
115 Filamentous Bacteria Microthrix parvicella
116 Filamentous Bacteria Microthrix parvicella Microthrix parvicella is associated with Animal and vegetable greases and fats Low F:M Colder temperatures
117 Filamentous Bacteria Nocardia
118 Filamentous Bacteria Nocardia This filament is strongly associated with Long MCRTs (sludge age) Greases, oils, fats, petroleum, rubber Warmer temperatures
119 Filamentous Bacteria About Nocardia Very little foaming Foaming No foaming
120 Filamentous Bacteria About Nocardia Foaming Less foaming No foaming
121 Nocardia Early Stage Nocardia
122 Nocardia Early Stage Nocardia
123 Nocardia Mid - Stage Nocardia
124 Nocardia Mid - Stage Nocardia
125 Nocardia Late Stage Nocardia
126 Nocardia Late Stage Nocardia
127 Nocardia Adult Stage Nocardia
128 Filamentous Bacteria Foaming Filaments Type 1863 Microthrix parvicella Nocardia
129 Foaming Filaments Neisser Stain Neisser (+) (bluish) Nostocoida limicola Type 0092 Neisser (+) Granules Microthrix Nocardia Neisser (-) (brownish) All others
130 Neisser Positive Granules Microthrix
131 Neisser Positive Granules Nocardia
132 Filament Control For low F:M or low food Add more food or get rid of some bugs For low DO Add more oxygen
133 Filament Control Excess greases, oils and fats Grease control For a long MCRT or Sludge age Shorten it Sulfur granules Pre-aeration, add oxygen
134 Most Important Keep a process chart of treatment system parameters Measure the parameters routinely and consistently Measure the parameters when the treatment system is working properly
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