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8/9/2019 Microbial Diversity and the Quality of Roof Harvested Rainwater
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Microbial Diversity and the Quality of
Roof-harvested Rainwater
C.A. Evans, P.J.C.A. Evans, P.J. CoombesCoombes, R.H., R.H. DunstanDunstan, T.L. Harrison, A. Martin,, T.L. Harrison, A. Martin,K.K. PigottPigott, J.N. Harris, J.N. Harris
School of Environmental and Life SciencesSchool of Environmental and Life Sciences
University of NewcastleUniversity of Newcastle
AustraliaAustralia
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Are nonAre non--pathogenic bacteria significant?pathogenic bacteria significant?
1. The treatment traineffect
Apparent improvements to the microbial and chemical qualityof water with passage through the collection system.
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Contaminatedrun-off
INLET
OUTLET
Improvedwater quality
Treatment train
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Are nonAre non--pathogenic bacteria significant?pathogenic bacteria significant?
1. The treatment traineffect
Apparent improvements to the microbial and chemical qualityof water with passage through the collection system.
2. Environmental bacteria comprise majority of organismspresent in rainwater tanks.
HPC >>> coliform counts - (orders of magnitude)
Bacterial composition influenced by wind patterns
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Wind direction/velocity & HPCWind direction/velocity & HPC
0
1000
2000
3000
4000
5000
6000
0 2 4 6 8
Wind velocity (m/sec)
HPC(cfu
/mL)
N
S
(r2 = 0.98, p
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Contaminatedrun-off
INLET
OUTLET
Improvedwater quality
Treatment train
Biofilm formation
Nutrient cycling
Chemical remediation
Competitive exclusionof pathogens
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OverviewOverview
1. Assessment of microbial diversity in 2 rainwater tanks.
Impact of hot water systems
Assessment of coliforms
2. Biofilm pilot studies
Biofilm development and the treatment train
In-vitro examination of Pb2+ uptake
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Microbial Diversity of 2 Rainwater TanksMicrobial Diversity of 2 Rainwater Tanks
Urban tank
Located in Newcastle (160kM Nth of Sydney) Duel 2.2kL galvanized iron tanks
Mains water top-up system
Hot water service set at 60C
Rural tank Located at Gulgong (250kM inland of Newcastle)
10kL aquaplate tank
Hot water service set at 50C.
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Table 1: Plate count and diversity summary
Tank Urban Rural
Sample type Cold Hot Cold Hot
Total no. of species identified 64 20 49 39
mean no. species/sample 23 3.4 8 1.2 16 2.3 14 4.3
Mean plate count (cfu/mL) 366 153 17 2.6 2263 1070 94 31
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Table 1: Plate count and diversity summary
Tank Urban Rural
Sample type Cold Hot Cold Hot
Total no. of species identified 64 20 49 39
mean no. species/sample 23 3.4 8 1.2 16 2.3 14 4.3
Mean plate count (cfu/mL) 366 153 17 2.6 2263 1070 94 31
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Table 1: Plate count and diversity summary
Tank Urban Rural
Sample type Cold Hot Cold Hot
Total no. of species identified 64 20 49 39
mean no. species/sample 23 3.4 8 1.2 16 2.3 14 4.3
Mean plate count (cfu/mL) 366 153 17 2.6 2263 1070 94 31
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Table 1: Plate count and diversity summary
Tank Urban Rural
Sample type Cold Hot Cold Hot
Total no. of species identified 64 20 49 39
mean no. species/sample 23 3.4 8 1.2 16 2.3 14 4.3
Mean plate count (cfu/mL) 366 153 17 2.6 2263 1070 94 31
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Table 1: Plate count and diversity summary
Tank Urban Rural
Sample type Cold Hot Cold Hot
Total no. of species identified 64 20 49 39
mean no. species/sample 23 3.4 8 1.2 16 2.3 14 4.3
Mean plate count (cfu/mL) 366 153 17 2.6 2263 1070 94 31
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Table 1: Plate count and diversity summary
Tank Urban Rural
Sample type Cold Hot Cold Hot
Total no. of species identified 64 20 49 39
mean no. species/sample 23 3.4 8 1.2 16 2.3 14 4.3
Mean plate count (cfu/mL) 366 153 17 2.6 2263 1070 94 31
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Table 1: Plate count and diversity summary
Tank Urban Rural
Sample type Cold Hot Cold Hot
Total no. of species identified 64 20 49 39
mean no. species/sample 23 3.4 8 1.2 16 2.3 14 4.3
Mean plate count (cfu/mL) 366 153 17 2.6 2263 1070 94 31
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Fig. 2: % composition of gram negative and gram
positive bacteria in samples from rainwater tanks.
0%10%
20%
30%
40%50%
60%
70%
80%
90%
100%
Ncle(cold) Gulg(cold) Ncle(hot) Gulg(hot)
Sample type
%c
om
position
G+ve
G-ve
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Fig. 1: Profiles of the Newcastle and Gulgong tanks basedon average counts of most abundant genera present.
0
50
100
150
200
250
300
350
400
450
Pseudom
onas
sp.
Serra
tiasp.
Delfti
asp
.
Janthino
bacte
rium
sp.
Yersini
asp
.
Bacillus
sp.
Unide
ntifie
d
Fungi
Herba
spirillum
sp.
Sphin
gomonas
sp.
Avge
count(cfu/ml
Gulg.
Ncle
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Faecal indicator presence and abundance relative to totalplate count
Sample type COLD HOT
Presence Avge % of Presence Avge % of
(% of samples) plate count (% of samples) plate count
E.coli 50 0.06 0.03 0 0
Enterococci 23 0.05 0.03 28 0.62 0.41
Faecal col. 77 0.52 0.27 28 0.62 0.41
Total col. 100 3.77 2.08 43 3.36 2.21
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BiofilmsBiofilms and the treatment trainand the treatment train
Location:Newcastle on east coast of Australia(approx. 160kM Nth of Sydney)
Tank Biofilm development Comment
A (G.I.) Advanced heavy slime layer with extensive clumping
B (Plastic) Moderate uniform slime layer around interior
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Table 3: Variation in bacterial counts between the tankwater column and tap outlet of tanks A and B.
Tank Sample type Bacterial count
E.coli HPC
(cfu/100ml) (cfu/mL)
A Tank water column 48 610Tap outlet 29 390
Improvement 34% 36%
B Tank water column 70 200
Tap outlet 60 160
Improvement 14% 20%
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Table 3: Variation in bacterial counts between the tankwater column and tap outlet of 2 tanks.
Tank Sample type Bacterial count
E.coli HPC
(cfu/100ml) (cfu/mL)
A Tank water column 48 610Tap outlet 29 390
Improvement 34% 36%
B Tank water column 70 200
Tap outlet 60 160
Improvement 14% 20%
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PbPb2+2+ uptake by inuptake by in--vitro cultivatedvitro cultivated biofilmbiofilm
E.colibiofilms cultivated in wells of perspex tissueculture plates for 72 hours.
Incubation broth replaced with sterile milli-Q waterspiked with Pb2+ (100ppb).
Pb2+ determined by ICP-MS at time = 0, 5, 10 and 20hours.
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Pb2+ uptake by in-vitro cultivated biofilm
40
50
60
70
80
90
100
110
0 5 10 15 20 25
Time (hrs)
Leadconc
.(ppb)
Biofilm
Control
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ConclusionsConclusions
Rainwater tanks harbour a wide diversity of microbial species.
Species composition may vary considerably between tanks atdifferent locations.
Species composition varies temporally at any given site.
Minimal evidence of faecal contamination.
Hot water systems deliver water of quality suitable for bathing.
Biofilms have capacity to chemically remediate water.
Class Order Family GenusPhylumSphingomonadaceae
Afi i
NovosphingobiumPorphyrobacter
SphingomonadalesB d hi bi
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Caulobacterales Caulobacteraceae Caulobacter
RhizobialesMethylobacteriaceae
Afipia
Methylobacterium
PorphyrobacterSphingomonas
BurkholderiaBurkholderiaceae
AcidovoraxComomonasDelftiaPolaromonasVariovorax
Comomonadaceae
Duganella
MassiliaNaxibacterHerbaspirillumJanthinobacterium
Oxalobacteriaceae
Ralstonia
Burkholderiales
AquaspirillumAquitaleaChromobacteriumIodobacterMicrovirgula
NesseriaceaeNesseriales
DechloromonasRhodocyclaceae
CitrobacterEnterobacter
ErwiniaEscherichiaHafniaKlebsiellaLeclerciaObesumbacteriumPantoeaProteusRahnellaRaoultellaSerratiaYersinia
Pseudomonadaceae
LuteibacterStenotrophomonasPseudoxanthomonas
Xanthomonadales
ClostridiumClostridia
Planococcaceae
BrevibacillusPaenibacillus
Paenibacillaceae
Staphylococcaceae
Bradyrhizobiaceae
RalstoniaceaeProteobacteria
Rhodocyclales
Enterobacteriales Enterobacteriaceae
Aeromonadales Aeromonadaceae Aeromonas
Chromatiales Chromatiaceae RheinheimeraMoraxellaceae Acinetobacter
MoraxellaPseudomonadalesPseudomonas
Xanthomonadaceae
ClostridiaceaeClostridiales
BacillaceaeBacillusExiguobacterium
Listeriaceae Listeria
Bacillales PlanococcusPlanomicrobiumSporasarcina
Staphylococcus
AerococcusAerococcaceae
CarnobacteriumDesemzia
Carnobacteriaceae
EnterococcusEnterococcaceae
LactococcusStreptococcaceae
Lactobacillales
Bacilli
FrigoribacteriumMicrobacterium
Microbacteriaceae
CorynebacteriumCorynebacteriaceae
ArthrobacterLeucobacterMicrococcus
Micrococcaceae
Gordoniaceae Gordonia
Mycobacteriaceae Mycobacterium
Nocardiaceae Rhodococcus
Streptomycetaceae Streptomyces
ActinomycetalesActinobacteria
Flavobacteriaceae ChryseobacteriumFlavobacterium
Flexibacteriaceae FlectobacillusHymenobacter
Sphingobacteriaceae Sphingobacterium
Flavobacteroidales
Sphingobacteriales
Flavobacteria
Sphingobacteria
Firmicutes
Actinobacteria
Bacteroidetes