Abstract

The design and performance characteristics of municipal drinking water systems can profoundly influence public health. To assess the operational attributes of an Accra, Ghana drinking water distribution system, high-throughput 454 pyrosequencing was employed to characterize its bacterial community composition. Samples from the waterworks and four household sources (one household tap and three polytank storage units) were analyzed within one of the Accra's distribution networks over a 4-month period. Samples provided between 9,059 and 20,076 reads (average = 13,056) that represented a broad range of bacterial diversity, including rare genera. Minimum Entropy Decomposition (MED) analysis showed that the sequences described four major assemblages. Assemblages 1 and 2 dominated the waterworks and household tap samples while polytank storage unit samples, with one exception, contained assemblages 3 or 4. The considerable bacterial taxonomic difference between different sources suggests that contamination and/or selective growth shapes bacterial community structures after treatment at the waterworks. Of particular interest are the major differences between the polytank samples following storage and the tap/waterworks samples, suggesting that water storage (stagnation) can select for unique microbial populations.

INTRODUCTION

Human health and development in every society depend upon access to safe drinking water. Despite the widespread use of vigorous sand filtration and disinfection of water supplies, contamination frequently leads to the transmission of microbial infections that cause serious illnesses and associated mortality worldwide (Craun et al. 2010). In Ghana, contaminated water accounts for about 70% of reported diseases (International Fact-Finding Mission 2002; WaterAid 2005). In many parts of the world, inconsistent supplies coupled with increased demand forces local storage of water (International Fact-Finding Mission 2002) that increases the potential to introduce other sources of water contamination. Thus, water quality depends upon the initial water treatment, the water transmission from the water plant to the point of use, and on-site water storage systems.

In Ghana, specifically Accra, residents face acute water supply shortages. An estimated one quarter of its residents receives a continuous municipal water supply, while approximately 35% have access only 2 days each week (WaterAid 2005). To insure access to sufficient amounts during the non-supply periods, most Ghana citizens must store water. Inhabitants of Accra employ various forms of water storage tanks, but black polyethylene tanks referred to as ‘polytanks’ have gained wide acceptance. The storage of water for hours or days encourages stagnancy that can result in microbial contamination of otherwise good quality drinking water (Ling et al. 2018).

Microbiological monitoring of water quality generally relies upon conventional culturing of indicator organisms. However, these culture-dependent approaches underestimate bacterial diversity and do not provide assessments of pathogenic species (Amann et al. 1995). Several reports describe the presence of microbes associated with waterborne disease outbreaks in treated drinking water that meet coliform regulations (Payment et al. 1991), and in some cases, bacterial biofilms in drinking water distribution systems (DWDS) persist despite heavy chlorine treatment (LeChevallier et al. 1996). As an alternative, sequence analyses of polymerase chain reaction (PCR) amplicons for 16S ribosomal ribonucleic acid (rRNA) genes can document the presence of viable and non-viable microbes in drinking water networks that conventional microbiological approaches would fail to detect (Williams et al. 2004; Eichler et al. 2006).

High-throughput sequencing of PCR amplicons from hypervariable regions in 16S rRNA genes provides an efficient means to profile bacterial communities including the detection of sequence reads from very low abundance taxa that constitute less than 0.1% of a microbial community (Tan et al. 2015). Taking advantage of this technology, this investigation characterized the bacterial diversity in treated water samples collected from the Weija waterworks in Accra, Ghana and its distribution system over a 4-month time frame (September 2009 to December 2009; rainy season to the beginning of the dry season). Analysis of thousands of amplicon sequences for the 16S rRNA gene hypervariable regions V5–V6 (such regions are bordered by constant sequences convenient for PCR amplification and have been used in multiple DNA sequence studies for microbial identification) provided information about the occurrence, relative abundance and diversity of bacterial microflora in this Ghanaian drinking water supply system and how the microbial communities differed based upon the source (waterworks versus direct domestic tap versus polytank storage units) and seasonal collection time of the water samples.

MATERIALS AND METHODS

Sampling sites

The Ghana Water Company Limited provides drinking water for people in western Accra through its operation of the Weija waterworks located 15 km west of Accra, Ghana (see Supplementary Figure 1). The warm and humid climate of the region varies from 24 °C in August to 32 °C in March with a mean temperature of 27 °C. A dry period extends from December through March, followed by a rainy period with two peaks in June and September (Karikari & Ansa-Asare 2006). The waterworks (GPS coordinates 5.575493 N 0.343162 W) is supplied by a dam impoundment on the river Densu that forms a reservoir 14 km long, 2.2 km wide, with a total surface area of 38 km2 and a mean depth of 5 m (Vanden Bossche & Bernacsek 1990). Treatment at the waterworks involves the abstraction of water from the intake into the treatment train: aeration, flocculation and coagulation using alum, sedimentation, rapid sand filtration and, finally, pH adjustment and chlorination with hydrated lime and chlorine gas (Addico et al. 2006). The distribution pipes supply about 64 million m3 of treated water annually to consumers (Van Rooijen et al. 2008) by gravity feed.

This study sampled treated water over a 4-month period in 2009 (30 September, 28 October, 25 November and 15 December) from the waterworks and four randomly selected households located in Dansoman, a suburb of western Accra, which is approximately 12 km from the waterworks (see Supplementary Figure 1). Water was sampled from households with either a direct tap supply or an emergency polyethylene storage tank (polytank), all connected to the waterworks supply network. The three polytanks that were analyzed in this work were labeled as A, B and C (as shown in Figure 1 and located in Supplementary Figure 1).

Figure 1

Sequence variant relative abundance versus the sample sources and the month of collection using MED analysis. The cluster heatmap was produced as part of the MED analysis (Eren et al. 2015), using the R packages vegan (Oksanen et al. 2016) and pheatmap (Kolde 2015). The underlying matrix was calculated using the Bray–Curtis dissimilarity metric (Bray & Curtis 1957). Clustering indicates similarity among samples (top dendrogram) and among ASV abundances (left dendrogram). The source [Weija waterworks/Mains (blue); Household tap (green), etc.] and month of the collection (September 2009; October 2009, etc.) of different samples are presented horizontally. One hundred and fifty-four (154) different ASVs were identified by the 16s sequence in the samples (listed in the same vertical order in Table 1). The % abundance of any given ASV in any of the 20 samples is indicated by the color of the sample bar (the color and number 0–16 refer to the sample abundance). These ASVs fell into four different MED microbial assemblages (1, 2, 3 and 4 on the left). The four assemblages were analyzed versus their distribution in regard to sources and sampling months (Figure 1); waterworks and tap samples are dominated by assemblages 1 and 2 and, with one exception (polytank A September sample), all polytank samples are dominated by assemblage 3 or 4.

Figure 1

Sequence variant relative abundance versus the sample sources and the month of collection using MED analysis. The cluster heatmap was produced as part of the MED analysis (Eren et al. 2015), using the R packages vegan (Oksanen et al. 2016) and pheatmap (Kolde 2015). The underlying matrix was calculated using the Bray–Curtis dissimilarity metric (Bray & Curtis 1957). Clustering indicates similarity among samples (top dendrogram) and among ASV abundances (left dendrogram). The source [Weija waterworks/Mains (blue); Household tap (green), etc.] and month of the collection (September 2009; October 2009, etc.) of different samples are presented horizontally. One hundred and fifty-four (154) different ASVs were identified by the 16s sequence in the samples (listed in the same vertical order in Table 1). The % abundance of any given ASV in any of the 20 samples is indicated by the color of the sample bar (the color and number 0–16 refer to the sample abundance). These ASVs fell into four different MED microbial assemblages (1, 2, 3 and 4 on the left). The four assemblages were analyzed versus their distribution in regard to sources and sampling months (Figure 1); waterworks and tap samples are dominated by assemblages 1 and 2 and, with one exception (polytank A September sample), all polytank samples are dominated by assemblage 3 or 4.

Table 1

Microbes found in a Ghanaian drinking water system. The 154 ASVs presents are indicated, with reference to Figure 1; that is, the listing is in the same order as presented in Figure 1 

ASV id ASV size September mains October mains November mains December mains September tap October tap November tap December tap September polytankA October polytankA November polytankA December polytankA September polytankB October polytankB November polytankB December polytankB September polytankC October polytankC November polytankC December polytankC Taxonomic classification 
2636 602 0.32 0.34 1.90 2.13 0.51 0.04 0.00 0.09 1.24 0.12 0.30 0.09 1.55 0.20 0.30 0.04 0.05 0.28 0.21 0.43 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Alcaligenaceae 
46 789 0.85 0.41 2.29 3.06 2.10 0.03 0.00 0.23 1.57 0.02 0.27 0.23 0.68 0.10 0.25 0.28 0.02 0.01 0.21 0.78 Bacteria; Proteobacteria; Gammaproteobacteria; Xanthomonadales 
2940 602 0.41 0.58 1.59 1.90 2.04 0.17 0.00 0.16 1.38 0.14 0.06 0.18 0.26 0.34 0.15 0.31 0.00 0.07 0.16 0.33 Bacteria; Proteobacteria; Deltaproteobacteria; Gammaproteobacteria; Xanthomonadales; Myxococcales 
2540 291 0.05 0.32 2.05 0.43 0.92 0.06 0.00 0.09 0.56 0.00 0.00 0.07 0.21 0.03 0.05 0.00 0.00 0.01 0.05 0.09 Bacteria; Planctomycetes; Phycisphaerae; Phycisphaerales; Phycisphaeraceae 
78 387 0.88 0.24 0.73 0.93 1.32 0.06 0.00 0.10 0.94 0.19 0.09 0.05 0.53 0.08 0.04 0.06 0.00 0.10 0.09 0.16 Bacteria; Proteobacteria; Gammaproteobacteria; Xanthomonadales 
2643 273 0.23 0.17 1.15 1.08 0.17 0.00 0.00 0.13 0.17 0.17 0.11 0.00 0.43 0.03 0.23 0.09 0.00 0.15 0.18 0.13 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Alcaligenaceae 
838 1,615 0.55 0.34 0.64 1.81 0.55 0.48 0.07 0.07 0.57 1.48 0.75 0.85 3.32 1.60 3.85 0.84 0.07 2.44 1.16 4.02 Bacteria; Actinobacteria; Actinobacteria; Frankiales; Sporichthyaceae 
155 847 0.93 0.21 0.44 0.63 0.38 0.06 0.02 0.03 0.42 0.14 0.38 0.11 0.85 2.29 1.40 0.16 0.05 0.44 1.83 2.73 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Burkholderiaceae; Polynucleobacter 
842 734 0.20 0.30 0.78 1.53 0.13 0.23 0.02 0.07 0.32 0.46 0.43 0.45 1.85 0.61 1.17 0.37 0.11 0.81 0.67 1.44 Bacteria; Actinobacteria; Actinobacteria; Frankiales; Sporichthyaceae 
232 1,204 0.85 1.09 7.07 2.24 0.29 0.48 0.26 0.44 0.74 0.32 0.47 0.16 0.94 0.52 0.78 0.22 0.04 1.13 0.65 1.28 Bacteria; Verrucomicrobia 
3042 727 2.26 0.51 0.75 2.35 0.74 0.35 0.05 0.07 0.53 0.22 0.09 0.18 0.66 0.13 0.30 0.19 0.00 1.05 0.47 1.12 Bacteria; Actinobacteria; Acidimicrobiia; Acidimicrobiales; Acidimicrobiaceae 
3041 576 1.03 0.94 0.86 2.11 0.42 0.46 0.00 0.19 0.28 0.22 0.11 0.14 0.17 0.25 0.11 0.22 0.00 0.58 0.30 1.21 Bacteria; Actinobacteria; Acidimicrobiia; Acidimicrobiales; Acidimicrobiaceae 
171 239 0.28 0.09 0.08 0.22 0.53 0.01 0.00 0.01 0.45 0.24 0.12 0.09 0.81 0.25 0.14 0.04 0.05 0.43 0.05 0.06 Bacteria; Verrucomicrobia; Spartobacteria; empty_order; empty_family; Chthoniobacter 
153 438 1.76 0.51 0.42 0.60 0.51 0.30 0.00 0.06 0.57 0.17 0.08 0.11 0.58 0.08 0.14 0.04 0.04 0.63 0.21 0.40 Bacteria; Actinobacteria; Acidimicrobiia; Acidimicrobiales; Acidimicrobiaceae 
69 313 2.26 0.06 0.08 0.15 0.23 0.04 0.02 0.04 0.39 0.27 0.08 0.11 0.45 0.02 0.08 0.01 0.00 0.54 0.07 0.17 Bacteria; Verrucomicrobia; Spartobacteria; empty_order; empty_family; Chthoniobacter 
840 208 0.07 0.00 0.10 0.24 0.06 0.06 0.00 0.03 0.07 0.20 0.14 0.20 0.40 0.13 0.40 0.13 0.00 0.17 0.23 0.67 Bacteria; Actinobacteria; Actinobacteria; Frankiales; Sporichthyaceae 
2471 351 0.00 0.11 0.25 1.12 0.31 0.01 0.05 0.04 0.32 0.09 0.17 0.07 0.47 0.29 0.10 0.10 0.00 0.29 0.07 1.78 Bacteria; Proteobacteria; Betaproteobacteria; Methylophilales; Methylophilaceae 
168 288 0.32 3.06 0.51 0.86 0.08 0.09 0.12 0.29 0.08 0.00 0.00 0.00 0.06 0.00 0.00 0.01 0.00 0.04 0.00 0.03 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
2286 340 0.30 1.03 1.83 1.36 0.17 0.03 0.00 0.47 0.13 0.02 0.17 0.20 0.02 0.00 0.10 0.01 0.00 0.01 0.05 0.07 Bacteria; Proteobacteria; Gammaproteobacteria; Legionellales; Legionellaceae; Legionella 
278 322 0.51 1.18 1.00 1.21 0.34 0.15 0.00 0.25 0.36 0.03 0.05 0.20 0.02 0.00 0.01 0.00 0.00 0.14 0.09 0.11 Bacteria; Proteobacteria; Gammaproteobacteria; Legionellales; Legionellaceae; Legionella 
3100 384 0.45 0.71 1.63 0.71 0.44 0.04 1.06 0.91 0.18 0.00 0.11 0.07 0.04 0.00 0.14 0.04 0.02 0.03 0.14 0.10 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
3166 310 0.03 1.82 1.83 0.45 0.06 0.01 0.90 0.35 0.15 0.00 0.00 0.00 0.02 0.00 0.01 0.03 0.00 0.03 0.04 0.09 Bacteria; Cyanobacteria 
3167 214 0.00 0.98 1.39 0.56 0.04 0.03 0.47 0.37 0.00 0.00 0.00 0.04 0.02 0.00 0.03 0.01 0.00 0.01 0.00 0.00 Bacteria; Cyanobacteria 
3023 434 0.10 0.71 2.75 1.94 0.04 0.00 0.00 0.38 0.01 0.02 0.08 0.09 0.04 0.07 0.14 0.06 0.00 0.01 0.21 0.80 Bacteria; Proteobacteria; Alphaproteobacteria 
2539 407 0.00 0.43 1.85 3.32 0.27 0.01 0.00 0.51 0.38 0.02 0.02 0.07 0.11 0.00 0.04 0.04 0.00 0.03 0.02 0.03 Bacteria; Planctomycetes; Phycisphaerae; Phycisphaerales; Phycisphaeraceae 
2542 386 0.02 0.34 2.37 1.98 0.27 0.00 0.00 0.42 0.27 0.43 0.08 0.05 0.15 0.05 0.03 0.01 0.00 0.00 0.05 0.16 Bacteria; Planctomycetes; Phycisphaerae; Phycisphaerales; Phycisphaeraceae 
133 357 0.05 0.17 2.10 2.09 0.57 0.01 0.00 0.23 0.53 0.00 0.02 0.02 0.09 0.02 0.10 0.01 0.04 0.00 0.04 0.07 Bacteria; Planctomycetes; Phycisphaerae; Phycisphaerales; Phycisphaeraceae 
2283 303 0.05 0.19 0.05 2.69 0.00 0.03 0.00 1.86 0.00 0.00 0.02 0.05 0.00 0.03 0.01 0.06 0.00 0.01 0.02 0.03 Bacteria; Proteobacteria; Gammaproteobacteria; Legionellales; Legionellaceae; Legionella 
189 232 0.00 0.04 0.92 2.48 0.00 0.00 0.00 0.53 0.01 0.02 0.00 0.02 0.00 0.00 0.00 0.03 0.00 0.00 0.00 0.03 Bacteria; Proteobacteria; Gammaproteobacteria; Methylococcales; Methylococcaceae; Methylocaldum 
681 267 0.00 0.11 1.29 2.43 0.02 0.03 0.00 0.64 0.01 0.00 0.02 0.00 0.00 0.00 0.01 0.01 0.00 0.00 0.00 0.07 Bacteria; Proteobacteria; Gammaproteobacteria; Methylococcales; Methylococcaceae; Methylocaldum 
1349 3,463 0.00 0.02 0.03 0.00 0.04 46.95 0.05 0.00 0.01 0.00 0.03 0.00 0.00 2.88 0.01 0.00 0.00 0.10 0.65 0.00 Bacteria; Proteobacteria; Betaproteobacteria; Neisseriales; Neisseriaceae; Vogesella 
108 455 0.23 0.73 0.03 0.00 0.46 1.97 0.00 0.10 0.43 0.00 0.00 0.11 0.02 3.25 0.00 0.01 0.04 0.03 0.04 0.00 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Comamonadaceae; Aquabacterium 
2946 400 0.02 0.00 0.12 0.07 0.23 4.18 0.09 0.00 0.01 0.05 0.99 0.00 0.06 0.07 0.00 0.07 0.00 0.00 0.02 0.03 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Comamonadaceae; Pelomonas 
1348 350 0.00 0.00 0.00 0.00 0.00 4.40 0.09 0.00 0.00 0.00 0.00 0.00 0.00 0.71 0.00 0.00 0.00 0.00 0.02 0.00 Bacteria; Proteobacteria; Betaproteobacteria; Neisseriales; Neisseriaceae; Vogesella 
57 841 0.00 0.00 0.25 0.00 0.00 9.28 2.44 0.07 0.00 0.00 0.08 0.00 0.30 0.42 0.23 0.03 0.02 0.04 0.14 0.01 Bacteria; Proteobacteria; Gammaproteobacteria; Pseudomonadales; Moraxellaceae; Acinetobacter 
64 630 0.07 0.06 0.63 0.09 0.00 4.24 4.47 0.10 0.03 0.02 0.27 0.02 0.02 0.51 0.40 0.00 0.04 0.00 0.14 0.01 Bacteria; Proteobacteria; Gammaproteobacteria; Aeromonadales; Aeromonadaceae; Aeromonas; caviae 
2232 1,456 0.00 0.00 0.02 0.02 2.96 0.87 21.31 2.53 1.03 0.07 0.05 0.05 0.02 0.10 0.25 0.45 0.05 0.04 0.23 0.10 Bacteria; Proteobacteria; Alphaproteobacteria; Rhodospirillales; Rhodospirillaceae 
1963 1,019 0.38 4.86 0.05 0.00 0.23 0.09 12.75 2.06 0.07 0.05 0.29 0.16 0.00 0.03 0.05 0.10 0.00 0.00 0.28 0.04 Bacteria; Proteobacteria; Alphaproteobacteria; Caulobacterales; Hyphomonadaceae 
2942 472 0.00 0.00 0.00 0.00 0.00 0.81 9.20 0.19 0.00 0.00 0.00 0.00 0.00 0.00 0.01 0.00 0.00 0.15 0.04 0.00 Bacteria; Proteobacteria; Betaproteobacteria; Rhodocyclales; Rhodocyclaceae; Dechloromonas; CL 
162 245 0.00 0.00 0.00 0.00 0.19 0.07 4.52 0.20 0.18 0.00 0.00 0.07 0.09 0.00 0.00 0.03 0.02 0.00 0.00 0.00 Bacteria; Proteobacteria; Deltaproteobacteria 
996 466 0.95 0.09 0.02 0.00 0.63 0.84 4.68 0.34 0.45 0.10 0.06 0.07 0.11 0.19 0.16 0.18 0.05 0.00 0.04 0.00 Bacteria; Proteobacteria; Alphaproteobacteria; Sphingomonadales; Sphingomonadaceae; Sandarakinorhabdus 
42 417 0.48 0.00 0.00 0.00 0.67 0.28 1.68 1.95 0.85 0.05 0.11 0.05 0.11 0.02 0.10 0.36 0.02 0.07 0.19 0.01 Bacteria; Proteobacteria; Alphaproteobacteria; Sphingomonadales 
980 439 0.81 0.02 0.03 0.00 0.32 0.33 2.10 2.21 0.36 0.07 0.21 0.11 0.06 0.10 0.07 0.27 0.00 0.03 0.37 0.03 Bacteria; Proteobacteria; Alphaproteobacteria; Rhizobiales 
160 434 0.88 0.49 0.44 0.45 0.38 0.30 1.68 0.20 0.17 0.03 1.17 0.34 0.11 0.17 0.19 0.13 0.00 0.21 0.12 0.16 Bacteria; Proteobacteria; Alphaproteobacteria; Rickettsiales; Rickettsiaceae; Rickettsia 
986 380 2.17 0.58 0.03 0.00 0.42 0.23 2.32 0.29 0.32 0.03 0.02 0.04 0.13 0.00 0.07 0.18 0.00 0.03 0.00 0.14 Bacteria; Proteobacteria; Alphaproteobacteria; Sphingomonadales; Sphingomonadaceae; Sphingomonas; alpina 
1731 348 0.07 0.00 0.10 0.21 0.59 0.44 0.66 0.12 1.33 0.09 0.05 0.04 0.21 0.00 0.15 0.15 0.05 0.29 0.84 0.30 Bacteria; Proteobacteria; Alphaproteobacteria; Rhizobiales 
3195 378 0.08 0.00 0.00 0.00 1.03 0.45 0.59 0.31 0.85 0.12 0.02 0.04 1.15 0.00 0.14 0.28 0.04 0.30 0.90 0.09 Bacteria; Proteobacteria; Alphaproteobacteria; Sphingomonadales; Sphingomonadaceae; Sphingomonas; BAC151 
102 264 0.00 0.00 0.02 0.11 0.21 0.15 0.54 0.25 0.38 0.20 0.53 0.07 0.15 0.03 0.14 0.31 0.90 0.06 0.18 0.18 Bacteria; Proteobacteria; Deltaproteobacteria 
2945 384 0.00 0.09 0.17 0.02 0.11 1.87 0.76 0.31 0.34 0.58 0.33 0.13 0.04 0.19 0.73 0.01 0.16 0.00 0.25 0.06 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Comamonadaceae; Roseateles; Mitsuaria 
2787 226 0.00 0.00 0.00 0.00 0.11 0.74 0.73 0.39 1.01 0.09 0.06 0.09 0.08 0.05 0.00 0.19 0.00 0.00 0.07 0.01 Bacteria; Proteobacteria; Alphaproteobacteria; Rickettsiales 
249 1,013 0.22 2.57 3.90 4.20 2.38 0.03 0.00 0.78 2.00 0.07 0.11 0.13 0.49 0.08 0.05 0.09 0.04 0.12 0.25 0.26 Bacteria; Planctomycetes; Phycisphaerae; Phycisphaerales; Phycisphaeraceae 
3144 1,121 2.34 3.45 3.22 2.69 1.56 0.07 0.19 0.47 1.78 0.26 0.05 0.11 1.06 0.13 0.00 0.07 0.07 1.10 0.35 0.48 Bacteria; Cyanobacteria; empty_class; Chroococcales; empty_family; Microcystis; aeruginosa 
3054 1,578 5.59 4.56 2.58 3.43 3.34 0.70 0.02 2.66 3.27 0.05 0.06 0.05 0.25 0.02 0.00 0.09 0.11 0.15 0.02 0.06 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
3072 1,678 3.67 5.95 4.19 1.36 1.93 1.26 2.81 2.87 2.06 0.31 0.29 0.36 0.28 0.08 0.25 0.25 0.20 0.47 0.46 0.37 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
1495 1,312 3.83 4.11 1.27 1.38 1.81 0.91 1.23 3.64 1.58 0.38 0.12 0.13 0.55 0.29 0.08 0.12 0.09 0.50 0.16 0.30 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
3131 1,088 7.80 1.01 0.54 0.78 2.86 0.22 0.38 1.07 2.59 0.05 0.03 0.04 0.34 0.07 0.04 0.01 0.07 0.14 0.07 0.10 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
926 4.13 1.58 0.71 1.18 2.90 0.13 0.05 1.56 2.46 0.07 0.06 0.04 0.25 0.07 0.05 0.06 0.07 0.08 0.04 0.07 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
3143 608 2.17 3.83 0.39 1.25 0.95 0.09 0.05 0.23 1.23 0.09 0.02 0.04 0.36 0.08 0.04 0.01 0.00 0.06 0.07 0.03 Bacteria; Cyanobacteria; empty_class; Chroococcales; empty_family; Microcystis; aeruginosa 
3145 569 1.43 2.80 0.71 1.51 0.61 0.07 0.07 0.22 1.37 0.07 0.02 0.09 0.43 0.24 0.00 0.01 0.02 0.11 0.11 0.18 Bacteria; Cyanobacteria; empty_class; Chroococcales; empty_family; Microcystis 
1457 907 4.41 2.29 1.02 0.76 1.72 0.45 0.61 0.98 1.61 0.19 0.09 0.07 0.66 0.17 0.08 0.07 0.07 0.12 0.11 0.11 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
1456 721 2.46 3.08 0.59 0.69 1.07 0.57 0.17 1.05 1.20 0.20 0.12 0.05 0.38 0.10 0.04 0.07 0.05 0.21 0.16 0.18 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
3112 604 1.82 1.95 1.07 0.80 0.92 0.30 0.35 1.26 1.09 0.10 0.05 0.07 0.06 0.07 0.04 0.07 0.02 0.17 0.11 0.03 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
979 3,169 1.33 0.83 0.08 0.00 9.27 2.52 2.10 21.26 8.61 0.22 0.08 0.22 0.43 0.32 0.14 1.50 0.09 0.21 0.21 0.17 Bacteria; Proteobacteria; Alphaproteobacteria; Rhizobiales 
50 2,679 1.09 3.32 0.05 0.00 4.21 3.48 6.08 18.06 3.30 0.34 0.14 0.22 1.19 0.25 0.29 0.87 0.33 0.25 0.47 0.07 Bacteria; Proteobacteria; Alphaproteobacteria; Caulobacterales; Hyphomonadaceae 
1192 3,486 5.21 12.82 4.68 7.91 2.27 1.36 3.74 12.53 2.67 1.13 0.41 0.47 0.75 0.20 0.37 0.75 0.11 0.80 0.81 1.36 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
3022 3,703 1.74 6.08 21.43 14.16 2.78 0.44 0.00 2.27 2.68 0.15 0.33 0.72 2.15 0.49 1.20 0.40 0.20 0.28 0.93 5.06 Bacteria; Proteobacteria; Alphaproteobacteria 
721 3,239 8.08 2.55 8.26 8.51 4.37 0.33 0.00 0.92 5.05 0.44 0.65 1.01 3.83 1.03 1.83 0.94 0.11 0.70 1.42 4.14 Bacteria; Proteobacteria; Alphaproteobacteria 
3084 444 1.89 1.54 0.31 0.26 0.88 0.23 0.12 0.80 0.87 0.03 0.03 0.02 0.23 0.00 0.00 0.00 0.02 0.25 0.00 0.09 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
3113 381 2.41 1.05 0.31 0.32 0.48 0.10 0.17 0.72 0.49 0.07 0.02 0.02 0.15 0.03 0.01 0.06 0.00 0.07 0.02 0.03 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
1649 420 2.06 2.29 0.36 0.54 0.38 0.10 0.07 0.04 0.50 0.14 0.00 0.00 0.30 0.13 0.08 0.04 0.04 0.19 0.09 0.11 Bacteria; Proteobacteria; Alphaproteobacteria; Rhodospirillales; Acetobacteraceae; Roseomonas; NA; SK_65; NA 
1455 350 0.93 1.84 0.03 0.34 0.61 0.30 0.07 0.78 0.38 0.10 0.03 0.02 0.09 0.07 0.01 0.10 0.02 0.26 0.00 0.09 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
3098 343 1.54 1.84 0.15 0.19 0.61 0.07 0.02 0.42 0.46 0.15 0.00 0.00 0.28 0.00 0.01 0.00 0.02 0.12 0.09 0.07 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
3132 266 1.39 0.45 0.20 0.35 0.61 0.12 0.07 0.23 0.70 0.03 0.02 0.02 0.11 0.03 0.01 0.03 0.04 0.01 0.04 0.01 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
3056 228 0.66 0.92 0.12 0.11 0.46 0.16 0.07 0.35 0.70 0.02 0.00 0.04 0.19 0.05 0.00 0.01 0.00 0.03 0.02 0.00 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
3086 225 0.80 0.71 0.07 0.24 0.38 0.16 0.12 0.42 0.38 0.07 0.00 0.02 0.11 0.02 0.01 0.06 0.02 0.18 0.00 0.06 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
984 564 0.30 0.15 0.00 0.00 2.76 0.61 0.45 1.43 2.56 0.10 0.03 0.02 0.26 0.12 0.11 0.06 0.00 0.08 0.04 0.03 Bacteria; Proteobacteria; Alphaproteobacteria; Rhizobiales 
2338 913 0.35 0.30 0.02 0.02 6.23 0.29 0.45 0.04 5.76 0.12 0.03 0.04 0.87 0.00 0.41 0.00 0.04 0.03 0.07 0.00 Bacteria; Proteobacteria; Alphaproteobacteria; Sphingomonadales; Sphingomonadaceae; Novosphingobium; aromaticivorans; NA; NA 
983 880 0.10 0.00 0.00 0.00 3.79 0.55 4.26 0.25 3.75 0.15 0.15 0.13 1.45 0.07 0.32 0.04 0.04 0.25 0.33 0.00 Bacteria; Proteobacteria; Alphaproteobacteria; Sphingomonadales; Sphingomonadaceae; Sphingomonas 
139 533 2.12 0.00 0.02 0.02 2.67 0.01 0.00 0.00 2.46 0.00 0.00 0.00 1.26 0.10 0.00 0.00 0.04 0.14 0.00 0.01 Bacteria; Chlamydiae; Chlamydiae; Chlamydiales; Chlamydiaceae 
74 624 2.24 0.41 0.20 0.11 3.15 0.03 0.02 0.01 2.46 0.14 0.00 0.02 1.06 0.08 0.03 0.03 0.09 0.18 0.02 0.20 Bacteria; Chlamydiae; Chlamydiae; Chlamydiales 
2943 462 1.56 0.04 0.03 0.09 2.36 0.32 0.14 0.18 1.66 0.29 0.02 0.02 0.53 0.03 0.08 0.06 0.13 0.03 0.09 0.04 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Comamonadaceae; Pelomonas 
2944 348 1.92 0.00 0.00 0.00 2.48 0.17 0.07 0.16 0.60 0.02 0.00 0.00 0.17 0.02 0.01 0.03 0.04 0.07 0.21 0.00 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Comamonadaceae; Inhella 
340 221 0.43 0.00 0.00 0.00 0.95 0.06 0.02 0.04 1.29 0.02 0.00 0.00 0.60 0.02 0.00 0.03 0.11 0.01 0.02 0.01 Bacteria; Proteobacteria; Deltaproteobacteria; Bdellovibrionales; Bacteriovoracaceae; Peredibacter 
1013 245 0.03 0.69 0.00 0.00 2.31 0.01 0.05 0.00 0.98 0.00 0.02 0.00 0.23 0.00 0.01 0.01 0.00 0.00 0.00 0.03 Bacteria; Proteobacteria; Alphaproteobacteria; Sphingomonadales; Sphingomonadaceae; Sphingomonas 
3239 2,214 0.00 0.11 0.05 0.04 0.10 0.04 0.09 0.03 0.06 0.00 0.02 0.11 19.51 9.20 0.49 0.04 0.94 0.12 8.71 0.04 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Comamonadaceae; Pseudorhodoferax 
3242 1,398 0.00 0.04 0.07 0.15 0.00 0.03 0.07 0.00 0.00 0.00 0.20 0.00 0.49 14.95 0.43 0.03 0.00 0.28 6.71 0.26 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Comamonadaceae 
1689 5,588 0.00 0.11 0.00 0.00 0.00 0.06 0.21 0.03 0.08 9.26 1.82 1.25 3.77 6.89 15.90 29.28 0.74 3.51 4.44 8.09 Bacteria; Bacteroidetes; Sphingobacteriia; Sphingobacteriales; Chitinophagaceae; Lacibacter 
1389 3,245 0.07 0.13 0.00 0.00 0.10 0.04 0.05 0.04 0.34 3.10 3.45 2.94 8.53 4.68 0.78 3.00 0.60 8.39 8.19 7.55 Bacteria; Bacteroidetes; Sphingobacteriia; Sphingobacteriales; Chitinophagaceae; Sediminibacterium 
45 2,210 0.00 0.00 0.00 0.00 0.02 0.00 0.00 0.00 0.32 2.40 3.12 1.95 3.74 4.90 2.45 2.50 2.80 3.54 7.34 0.97 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Oxalobacteraceae; Paucimonas 
3206 3,202 0.48 0.11 0.02 0.00 0.06 0.03 0.05 0.07 0.52 7.59 2.55 2.29 10.24 5.76 4.92 4.84 0.85 2.80 7.32 2.06 Bacteria; Proteobacteria; Alphaproteobacteria; Sphingomonadales; Sphingomonadaceae; Novosphingobium; FW − 6 
2570 1,585 0.05 0.00 0.02 0.02 0.11 0.00 0.02 0.00 0.08 0.05 0.17 0.00 0.19 3.35 8.56 7.57 0.00 0.14 2.81 0.65 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Comamonadaceae; Pseudorhodoferax 
128 1,137 0.00 0.00 0.00 0.00 0.00 0.04 0.00 0.00 0.04 0.12 0.05 0.04 0.98 1.69 4.92 3.61 0.05 3.79 1.46 0.10 Bacteria; Proteobacteria; Deltaproteobacteria 
1852 1,066 0.00 0.00 0.00 0.00 0.00 0.07 0.00 0.00 1.94 0.14 0.29 0.00 0.15 2.02 5.22 3.48 0.00 0.70 1.84 0.03 Bacteria; Proteobacteria; Alphaproteobacteria; Caulobacterales; Caulobacteraceae; Caulobacter; Brevundimonas 
269 956 0.00 0.00 0.02 0.04 0.02 0.00 0.02 0.00 0.04 0.03 0.00 0.00 3.58 4.50 2.35 0.84 0.20 0.22 3.86 0.21 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Comamonadaceae; Polaromonas 
123 597 0.00 0.00 0.03 0.00 0.13 0.59 0.09 0.00 0.07 0.00 0.00 0.02 1.09 2.65 2.80 1.15 0.02 0.06 0.58 0.04 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Comamonadaceae; NA; Piscinibacter 
924 668 0.05 0.21 0.00 0.00 0.00 0.01 0.09 0.00 0.10 0.03 0.11 0.02 0.45 1.58 2.75 1.20 0.02 0.55 1.55 1.51 Bacteria; Proteobacteria; Betaproteobacteria 
1650 434 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.01 0.00 6.15 Bacteria; Proteobacteria; Alphaproteobacteria; Rhodospirillales; Acetobacteraceae; Roseomonas; gilardii; NA; NA; subsp._rosea 
146 998 0.00 0.00 0.00 0.00 0.06 0.00 0.00 0.00 0.07 0.68 0.64 0.42 0.02 1.77 0.16 0.82 0.11 0.12 7.03 4.22 Bacteria; Verrucomicrobia 
145 643 0.12 0.00 0.07 0.00 0.06 0.09 0.09 0.06 0.13 0.03 0.24 0.18 0.06 0.13 0.99 1.75 0.23 0.18 3.58 2.10 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae; Planctomyces 
2148 351 0.00 0.00 0.02 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.02 0.07 0.99 0.28 0.00 0.32 3.90 0.13 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae; Planctomyces 
2573 344 0.00 0.02 0.02 0.04 0.11 0.62 0.26 0.00 0.10 0.02 0.23 0.02 0.98 1.40 0.82 0.01 0.16 0.06 0.79 0.03 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Comamonadaceae; Acidovorax 
1994 428 0.00 0.00 0.00 0.00 0.04 0.00 0.00 0.00 0.06 1.47 0.14 0.40 0.26 0.86 1.51 0.31 0.00 0.29 1.26 0.23 Bacteria; Proteobacteria; Alphaproteobacteria; Rhodospirillales; Acetobacteraceae; Gluconacetobacter; diazotrophicus; ICB557; NA 
1151 296 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.34 0.66 1.11 1.53 0.00 0.02 0.00 1.12 0.00 Bacteria; Cyanobacteria; Chloroplast 
1849 270 0.02 0.00 0.00 0.00 0.00 0.01 0.00 0.00 0.00 0.00 0.00 0.00 1.28 0.71 1.55 0.21 0.00 0.00 0.54 0.00 Bacteria; Proteobacteria; Alphaproteobacteria; Rhodobacterales; Rhodobacteraceae; Rhodobacter 
1734 337 0.00 0.00 0.00 0.00 0.02 0.07 0.00 0.03 0.11 0.46 0.20 0.09 1.15 0.37 0.40 1.83 0.13 0.14 0.46 0.03 Bacteria; Proteobacteria; Alphaproteobacteria; Caulobacterales; Caulobacteraceae; Brevundimonas 
680 328 0.00 0.04 0.02 0.00 0.00 0.00 0.02 0.00 0.22 0.14 0.03 0.00 1.81 0.15 0.25 1.33 0.54 0.19 0.12 0.50 Bacteria; Proteobacteria; Gammaproteobacteria; Xanthomonadales; Xanthomonadaceae; Thermomonas; R039N 
2685 228 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.06 0.03 0.44 0.11 0.00 0.54 0.12 0.27 0.02 0.14 1.98 0.06 Bacteria; Bacteroidetes; Sphingobacteriia; Sphingobacteriales 
1265 206 0.00 0.00 0.00 0.06 0.06 0.03 0.00 0.01 0.17 0.09 0.05 0.00 0.15 0.44 0.43 0.25 0.00 0.61 0.76 0.11 Bacteria; Proteobacteria; Alphaproteobacteria; Caulobacterales; Hyphomonadaceae; Hirschia 
1956 195 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.56 0.06 0.27 0.09 0.20 1.04 0.24 0.00 0.26 0.07 0.16 Bacteria; Proteobacteria; Alphaproteobacteria; Rhizobiales; Hyphomicrobiaceae; Hyphomicrobium 
3223 194 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.32 0.26 0.05 0.08 0.22 0.43 0.00 0.02 1.35 0.00 0.11 Bacteria; Proteobacteria; Alphaproteobacteria; Rickettsiales 
2470 262 0.00 0.00 0.00 0.00 0.06 0.00 0.14 0.04 0.39 0.03 0.08 1.01 0.04 0.00 0.27 0.07 0.54 1.24 0.21 0.00 Bacteria; Proteobacteria; Betaproteobacteria; Nitrosomonadales; Nitrosomonadaceae 
14 347 0.00 0.00 0.00 0.00 0.00 0.00 0.02 0.01 0.00 0.39 0.43 0.99 0.13 0.13 0.60 0.34 0.07 0.61 0.07 1.51 Bacteria; Acidobacteria 
621 499 0.00 0.00 0.03 0.07 0.00 0.00 0.02 0.00 0.03 0.73 1.38 1.08 0.00 0.39 0.65 0.60 0.23 0.48 0.11 1.88 Bacteria; Proteobacteria; Betaproteobacteria; Nitrosomonadales; Nitrosomonadaceae 
76 536 0.00 0.02 0.00 0.02 0.00 0.00 0.00 0.01 0.36 0.70 0.73 0.87 0.13 0.37 0.48 0.49 0.51 1.50 1.33 0.85 Bacteria; Proteobacteria; Deltaproteobacteria; Myxococcales 
182 426 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.01 0.48 0.90 2.17 0.02 0.32 0.26 0.85 0.67 0.21 0.37 0.70 Bacteria; Verrucomicrobia 
2778 291 0.00 0.00 0.00 0.00 0.10 0.00 0.07 0.04 0.11 0.14 0.55 0.88 0.02 0.19 0.37 0.45 0.49 0.40 0.18 0.63 Bacteria; Proteobacteria; Alphaproteobacteria; Rhodospirillales; Rhodospirillaceae 
1961 955 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.15 2.24 0.98 0.00 2.14 0.55 0.16 0.13 4.51 0.14 3.20 Bacteria; Proteobacteria; Alphaproteobacteria; Rhodobacterales; Rhodobacteraceae; Pseudorhodobacter; Rhodobacter; NA; 16 − 62; NA 
2683 904 0.00 0.00 0.03 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.03 1.32 0.11 0.59 0.80 0.70 0.00 3.76 0.00 5.80 Bacteria; Bacteroidetes; Sphingobacteriia; Sphingobacteriales 
285 538 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.06 0.29 0.00 0.78 0.07 0.49 0.00 3.38 0.44 2.33 Bacteria; Proteobacteria; Alphaproteobacteria; Sphingomonadales; Sphingomonadaceae; Zymomonas 
15 678 0.00 0.00 0.15 0.09 0.04 0.00 0.00 0.04 0.00 0.00 0.05 0.20 0.00 0.22 1.54 2.64 0.00 2.98 0.04 1.79 Bacteria; Proteobacteria; Betaproteobacteria; Methylophilales; Methylophilaceae 
1957 583 0.03 0.04 0.00 0.02 0.00 0.00 0.00 0.04 0.01 0.92 0.47 0.47 0.06 0.39 1.13 1.23 0.20 1.89 0.12 1.68 Bacteria; Proteobacteria; Alphaproteobacteria; Rhodospirillales; empty_family; Reyranella 
2334 772 0.00 0.00 0.02 0.00 0.00 0.00 0.00 0.00 0.01 1.26 0.67 1.54 0.17 0.54 1.98 0.55 0.00 2.76 0.07 2.00 Bacteria; Proteobacteria; Alphaproteobacteria; Rickettsiales 
3219 714 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.01 1.09 0.64 1.44 0.08 0.35 1.03 0.78 0.02 3.03 0.11 2.10 Bacteria; Proteobacteria; Alphaproteobacteria; Rickettsiales 
110 1,531 0.00 0.00 0.02 0.02 0.17 0.00 0.05 0.04 0.01 0.97 1.89 4.55 0.02 0.08 0.04 0.19 18.16 0.47 0.04 0.26 Bacteria; Verrucomicrobia; Opitutae; Opitutales; Opitutaceae; Opitutus 
1242 1,440 0.00 0.00 0.00 0.00 0.04 0.01 0.00 0.00 0.06 0.00 0.00 0.00 0.06 0.00 0.00 0.01 25.73 0.00 0.09 0.00 Bacteria; Chlamydiae; Chlamydiae; Chlamydiales; Simkaniaceae; Candidatus_Rhabdochlamydia 
94 612 0.00 0.00 0.00 0.00 0.02 0.01 0.09 0.01 0.24 0.44 0.99 1.08 0.02 0.13 0.45 0.33 3.96 1.46 0.07 0.63 Bacteria; Gemmatimonadetes; Gemmatimonadetes; Gemmatimonadales; Gemmatimonadaceae 
2145 603 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.01 0.00 0.44 1.28 0.56 0.00 0.00 0.00 0.16 5.62 1.34 0.04 0.57 Bacteria; Proteobacteria; Alphaproteobacteria; Rhodospirillales 
16 305 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.03 0.00 0.56 1.05 1.14 0.00 0.00 0.00 0.00 2.48 0.01 0.00 0.00 Bacteria; Nitrospirae; Nitrospira 
180 250 0.00 0.00 0.00 0.00 0.00 0.00 0.05 0.01 0.01 0.12 1.05 0.00 0.00 0.02 0.03 0.01 2.98 0.00 0.02 0.00 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae; Planctomyces 
2895 716 0.00 0.00 0.02 0.00 0.00 0.00 0.00 0.03 0.21 0.00 0.00 0.02 0.02 0.94 0.22 5.76 3.99 0.21 0.00 0.06 Bacteria; Verrucomicrobia 
32 435 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 7.86 0.00 0.00 0.00 Bacteria; Verrucomicrobia 
82 304 0.00 0.02 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.02 0.02 0.02 0.00 0.00 0.00 0.00 5.42 0.00 0.00 0.00 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae; Gemmata 
2896 505 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.11 0.02 0.11 6.39 0.04 0.00 0.19 0.12 0.22 0.04 0.14 1.25 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae; Planctomyces 
124 370 0.00 0.00 0.00 0.00 0.00 0.00 0.02 0.01 0.00 0.36 1.14 3.67 0.02 0.00 0.00 0.00 0.47 0.54 0.00 0.04 Bacteria; Proteobacteria; Betaproteobacteria; Nitrosomonadales; Nitrosomonadaceae 
1733 291 0.00 0.06 0.00 0.00 0.04 0.01 0.17 0.57 0.06 0.19 0.88 2.67 0.04 0.00 0.01 0.04 0.14 0.01 0.02 0.03 Bacteria; Proteobacteria; Alphaproteobacteria; Rhodospirillales; Rhodospirillaceae 
126 224 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.01 0.56 0.82 1.14 0.00 0.00 0.15 0.04 0.38 0.50 0.00 0.04 Bacteria; Acidobacteria 
65 360 0.03 0.02 0.02 0.06 0.00 0.00 0.05 0.01 0.06 0.87 1.58 2.02 0.02 0.00 0.15 0.04 0.23 0.08 0.07 0.58 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
113 216 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.63 1.49 1.46 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 Bacteria 
97 255 0.00 0.00 0.00 0.02 0.04 0.03 0.02 0.04 0.01 0.77 1.51 1.32 0.00 0.00 0.03 0.01 0.36 0.04 0.04 0.00 Bacteria; Proteobacteria; Alphaproteobacteria; Rhodospirillales 
620 410 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 1.77 2.27 1.79 0.00 0.02 0.01 0.01 0.04 0.73 0.00 0.00 Bacteria; Proteobacteria; Betaproteobacteria; Nitrosomonadales; Nitrosomonadaceae 
3196 572 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.03 2.05 3.22 2.33 0.02 0.07 0.10 0.00 1.66 0.03 0.00 0.04 Bacteria; Proteobacteria; Alphaproteobacteria; Rickettsiales 
1735 418 0.00 0.00 0.00 0.00 0.00 0.01 0.05 0.00 0.03 2.17 1.40 2.62 0.00 0.00 0.00 0.00 0.47 0.23 0.04 0.06 Bacteria; Proteobacteria; Alphaproteobacteria; Rhodospirillales; empty_family; Reyranella 
3186 521 0.00 0.02 0.00 0.00 0.00 0.00 0.00 0.00 0.20 1.82 0.70 1.34 0.15 0.12 1.61 0.16 0.23 1.52 0.04 0.16 Bacteria; Planctomycetes; Phycisphaerae; Phycisphaerales; Phycisphaeraceae 
1149 287 0.00 0.00 0.02 0.07 0.00 0.01 0.02 0.06 0.41 1.31 0.21 2.15 0.00 0.05 0.01 0.16 0.29 0.00 0.05 0.04 Bacteria 
2146 522 0.00 0.00 0.00 0.00 0.13 0.00 0.00 0.04 0.22 6.39 0.49 0.27 0.21 0.00 0.01 0.03 0.45 0.47 0.02 0.00 Bacteria; Proteobacteria; Alphaproteobacteria; Caulobacterales; Caulobacteraceae; Caulobacter; Es3; MDB1–11; NA 
24 315 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 5.34 0.00 0.00 0.00 0.00 0.00 0.00 0.04 0.00 0.00 0.00 Bacteria; Verrucomicrobia 
261 290 0.10 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 3.07 0.04 0.02 0.00 0.00 0.00 0.00 1.09 0.00 0.00 Bacteria 
311 470 0.02 0.00 0.00 0.00 0.11 0.04 0.33 0.07 0.80 0.26 4.23 1.28 0.02 0.02 0.01 0.04 0.07 0.03 0.09 0.04 Bacteria; Proteobacteria; Deltaproteobacteria 
163 755 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.02 5.81 2.82 0.00 0.00 0.00 0.00 0.00 2.95 0.00 0.03 Bacteria; Bacteroidetes; Sphingobacteriia; Sphingobacteriales 
2231 845 0.02 0.00 0.00 0.19 0.02 0.03 0.00 0.01 0.01 0.80 9.35 2.00 0.00 0.00 0.00 0.22 0.05 0.45 0.02 0.06 Bacteria; Proteobacteria 
1386 2,909 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 20.34 10.55 12.66 0.00 0.03 0.04 0.01 0.07 4.05 0.00 0.28 Bacteria; Bacteroidetes; Sphingobacteriia; Sphingobacteriales; Chitinophagaceae; Ferruginibacter 
925 1,808 0.00 0.00 0.02 0.02 0.00 0.00 0.00 0.00 0.06 3.26 7.01 5.24 0.02 0.07 2.35 3.33 2.42 2.98 0.09 1.52 Bacteria; Proteobacteria; Betaproteobacteria 
2143 1,324 0.10 0.02 0.12 0.28 0.21 0.04 0.07 0.07 0.45 1.36 2.54 2.35 0.11 1.31 1.84 1.36 0.78 5.64 0.97 0.70 Bacteria; Proteobacteria; Alphaproteobacteria; Rhizobiales 
3184 1,131 0.00 0.06 0.03 0.02 0.02 0.00 0.00 0.03 0.55 2.63 1.72 3.88 0.43 0.25 2.76 1.29 0.98 2.49 0.07 0.53 Bacteria; Planctomycetes; Phycisphaerae; Phycisphaerales; Phycisphaeraceae 
ASV id ASV size September mains October mains November mains December mains September tap October tap November tap December tap September polytankA October polytankA November polytankA December polytankA September polytankB October polytankB November polytankB December polytankB September polytankC October polytankC November polytankC December polytankC Taxonomic classification 
2636 602 0.32 0.34 1.90 2.13 0.51 0.04 0.00 0.09 1.24 0.12 0.30 0.09 1.55 0.20 0.30 0.04 0.05 0.28 0.21 0.43 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Alcaligenaceae 
46 789 0.85 0.41 2.29 3.06 2.10 0.03 0.00 0.23 1.57 0.02 0.27 0.23 0.68 0.10 0.25 0.28 0.02 0.01 0.21 0.78 Bacteria; Proteobacteria; Gammaproteobacteria; Xanthomonadales 
2940 602 0.41 0.58 1.59 1.90 2.04 0.17 0.00 0.16 1.38 0.14 0.06 0.18 0.26 0.34 0.15 0.31 0.00 0.07 0.16 0.33 Bacteria; Proteobacteria; Deltaproteobacteria; Gammaproteobacteria; Xanthomonadales; Myxococcales 
2540 291 0.05 0.32 2.05 0.43 0.92 0.06 0.00 0.09 0.56 0.00 0.00 0.07 0.21 0.03 0.05 0.00 0.00 0.01 0.05 0.09 Bacteria; Planctomycetes; Phycisphaerae; Phycisphaerales; Phycisphaeraceae 
78 387 0.88 0.24 0.73 0.93 1.32 0.06 0.00 0.10 0.94 0.19 0.09 0.05 0.53 0.08 0.04 0.06 0.00 0.10 0.09 0.16 Bacteria; Proteobacteria; Gammaproteobacteria; Xanthomonadales 
2643 273 0.23 0.17 1.15 1.08 0.17 0.00 0.00 0.13 0.17 0.17 0.11 0.00 0.43 0.03 0.23 0.09 0.00 0.15 0.18 0.13 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Alcaligenaceae 
838 1,615 0.55 0.34 0.64 1.81 0.55 0.48 0.07 0.07 0.57 1.48 0.75 0.85 3.32 1.60 3.85 0.84 0.07 2.44 1.16 4.02 Bacteria; Actinobacteria; Actinobacteria; Frankiales; Sporichthyaceae 
155 847 0.93 0.21 0.44 0.63 0.38 0.06 0.02 0.03 0.42 0.14 0.38 0.11 0.85 2.29 1.40 0.16 0.05 0.44 1.83 2.73 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Burkholderiaceae; Polynucleobacter 
842 734 0.20 0.30 0.78 1.53 0.13 0.23 0.02 0.07 0.32 0.46 0.43 0.45 1.85 0.61 1.17 0.37 0.11 0.81 0.67 1.44 Bacteria; Actinobacteria; Actinobacteria; Frankiales; Sporichthyaceae 
232 1,204 0.85 1.09 7.07 2.24 0.29 0.48 0.26 0.44 0.74 0.32 0.47 0.16 0.94 0.52 0.78 0.22 0.04 1.13 0.65 1.28 Bacteria; Verrucomicrobia 
3042 727 2.26 0.51 0.75 2.35 0.74 0.35 0.05 0.07 0.53 0.22 0.09 0.18 0.66 0.13 0.30 0.19 0.00 1.05 0.47 1.12 Bacteria; Actinobacteria; Acidimicrobiia; Acidimicrobiales; Acidimicrobiaceae 
3041 576 1.03 0.94 0.86 2.11 0.42 0.46 0.00 0.19 0.28 0.22 0.11 0.14 0.17 0.25 0.11 0.22 0.00 0.58 0.30 1.21 Bacteria; Actinobacteria; Acidimicrobiia; Acidimicrobiales; Acidimicrobiaceae 
171 239 0.28 0.09 0.08 0.22 0.53 0.01 0.00 0.01 0.45 0.24 0.12 0.09 0.81 0.25 0.14 0.04 0.05 0.43 0.05 0.06 Bacteria; Verrucomicrobia; Spartobacteria; empty_order; empty_family; Chthoniobacter 
153 438 1.76 0.51 0.42 0.60 0.51 0.30 0.00 0.06 0.57 0.17 0.08 0.11 0.58 0.08 0.14 0.04 0.04 0.63 0.21 0.40 Bacteria; Actinobacteria; Acidimicrobiia; Acidimicrobiales; Acidimicrobiaceae 
69 313 2.26 0.06 0.08 0.15 0.23 0.04 0.02 0.04 0.39 0.27 0.08 0.11 0.45 0.02 0.08 0.01 0.00 0.54 0.07 0.17 Bacteria; Verrucomicrobia; Spartobacteria; empty_order; empty_family; Chthoniobacter 
840 208 0.07 0.00 0.10 0.24 0.06 0.06 0.00 0.03 0.07 0.20 0.14 0.20 0.40 0.13 0.40 0.13 0.00 0.17 0.23 0.67 Bacteria; Actinobacteria; Actinobacteria; Frankiales; Sporichthyaceae 
2471 351 0.00 0.11 0.25 1.12 0.31 0.01 0.05 0.04 0.32 0.09 0.17 0.07 0.47 0.29 0.10 0.10 0.00 0.29 0.07 1.78 Bacteria; Proteobacteria; Betaproteobacteria; Methylophilales; Methylophilaceae 
168 288 0.32 3.06 0.51 0.86 0.08 0.09 0.12 0.29 0.08 0.00 0.00 0.00 0.06 0.00 0.00 0.01 0.00 0.04 0.00 0.03 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
2286 340 0.30 1.03 1.83 1.36 0.17 0.03 0.00 0.47 0.13 0.02 0.17 0.20 0.02 0.00 0.10 0.01 0.00 0.01 0.05 0.07 Bacteria; Proteobacteria; Gammaproteobacteria; Legionellales; Legionellaceae; Legionella 
278 322 0.51 1.18 1.00 1.21 0.34 0.15 0.00 0.25 0.36 0.03 0.05 0.20 0.02 0.00 0.01 0.00 0.00 0.14 0.09 0.11 Bacteria; Proteobacteria; Gammaproteobacteria; Legionellales; Legionellaceae; Legionella 
3100 384 0.45 0.71 1.63 0.71 0.44 0.04 1.06 0.91 0.18 0.00 0.11 0.07 0.04 0.00 0.14 0.04 0.02 0.03 0.14 0.10 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
3166 310 0.03 1.82 1.83 0.45 0.06 0.01 0.90 0.35 0.15 0.00 0.00 0.00 0.02 0.00 0.01 0.03 0.00 0.03 0.04 0.09 Bacteria; Cyanobacteria 
3167 214 0.00 0.98 1.39 0.56 0.04 0.03 0.47 0.37 0.00 0.00 0.00 0.04 0.02 0.00 0.03 0.01 0.00 0.01 0.00 0.00 Bacteria; Cyanobacteria 
3023 434 0.10 0.71 2.75 1.94 0.04 0.00 0.00 0.38 0.01 0.02 0.08 0.09 0.04 0.07 0.14 0.06 0.00 0.01 0.21 0.80 Bacteria; Proteobacteria; Alphaproteobacteria 
2539 407 0.00 0.43 1.85 3.32 0.27 0.01 0.00 0.51 0.38 0.02 0.02 0.07 0.11 0.00 0.04 0.04 0.00 0.03 0.02 0.03 Bacteria; Planctomycetes; Phycisphaerae; Phycisphaerales; Phycisphaeraceae 
2542 386 0.02 0.34 2.37 1.98 0.27 0.00 0.00 0.42 0.27 0.43 0.08 0.05 0.15 0.05 0.03 0.01 0.00 0.00 0.05 0.16 Bacteria; Planctomycetes; Phycisphaerae; Phycisphaerales; Phycisphaeraceae 
133 357 0.05 0.17 2.10 2.09 0.57 0.01 0.00 0.23 0.53 0.00 0.02 0.02 0.09 0.02 0.10 0.01 0.04 0.00 0.04 0.07 Bacteria; Planctomycetes; Phycisphaerae; Phycisphaerales; Phycisphaeraceae 
2283 303 0.05 0.19 0.05 2.69 0.00 0.03 0.00 1.86 0.00 0.00 0.02 0.05 0.00 0.03 0.01 0.06 0.00 0.01 0.02 0.03 Bacteria; Proteobacteria; Gammaproteobacteria; Legionellales; Legionellaceae; Legionella 
189 232 0.00 0.04 0.92 2.48 0.00 0.00 0.00 0.53 0.01 0.02 0.00 0.02 0.00 0.00 0.00 0.03 0.00 0.00 0.00 0.03 Bacteria; Proteobacteria; Gammaproteobacteria; Methylococcales; Methylococcaceae; Methylocaldum 
681 267 0.00 0.11 1.29 2.43 0.02 0.03 0.00 0.64 0.01 0.00 0.02 0.00 0.00 0.00 0.01 0.01 0.00 0.00 0.00 0.07 Bacteria; Proteobacteria; Gammaproteobacteria; Methylococcales; Methylococcaceae; Methylocaldum 
1349 3,463 0.00 0.02 0.03 0.00 0.04 46.95 0.05 0.00 0.01 0.00 0.03 0.00 0.00 2.88 0.01 0.00 0.00 0.10 0.65 0.00 Bacteria; Proteobacteria; Betaproteobacteria; Neisseriales; Neisseriaceae; Vogesella 
108 455 0.23 0.73 0.03 0.00 0.46 1.97 0.00 0.10 0.43 0.00 0.00 0.11 0.02 3.25 0.00 0.01 0.04 0.03 0.04 0.00 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Comamonadaceae; Aquabacterium 
2946 400 0.02 0.00 0.12 0.07 0.23 4.18 0.09 0.00 0.01 0.05 0.99 0.00 0.06 0.07 0.00 0.07 0.00 0.00 0.02 0.03 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Comamonadaceae; Pelomonas 
1348 350 0.00 0.00 0.00 0.00 0.00 4.40 0.09 0.00 0.00 0.00 0.00 0.00 0.00 0.71 0.00 0.00 0.00 0.00 0.02 0.00 Bacteria; Proteobacteria; Betaproteobacteria; Neisseriales; Neisseriaceae; Vogesella 
57 841 0.00 0.00 0.25 0.00 0.00 9.28 2.44 0.07 0.00 0.00 0.08 0.00 0.30 0.42 0.23 0.03 0.02 0.04 0.14 0.01 Bacteria; Proteobacteria; Gammaproteobacteria; Pseudomonadales; Moraxellaceae; Acinetobacter 
64 630 0.07 0.06 0.63 0.09 0.00 4.24 4.47 0.10 0.03 0.02 0.27 0.02 0.02 0.51 0.40 0.00 0.04 0.00 0.14 0.01 Bacteria; Proteobacteria; Gammaproteobacteria; Aeromonadales; Aeromonadaceae; Aeromonas; caviae 
2232 1,456 0.00 0.00 0.02 0.02 2.96 0.87 21.31 2.53 1.03 0.07 0.05 0.05 0.02 0.10 0.25 0.45 0.05 0.04 0.23 0.10 Bacteria; Proteobacteria; Alphaproteobacteria; Rhodospirillales; Rhodospirillaceae 
1963 1,019 0.38 4.86 0.05 0.00 0.23 0.09 12.75 2.06 0.07 0.05 0.29 0.16 0.00 0.03 0.05 0.10 0.00 0.00 0.28 0.04 Bacteria; Proteobacteria; Alphaproteobacteria; Caulobacterales; Hyphomonadaceae 
2942 472 0.00 0.00 0.00 0.00 0.00 0.81 9.20 0.19 0.00 0.00 0.00 0.00 0.00 0.00 0.01 0.00 0.00 0.15 0.04 0.00 Bacteria; Proteobacteria; Betaproteobacteria; Rhodocyclales; Rhodocyclaceae; Dechloromonas; CL 
162 245 0.00 0.00 0.00 0.00 0.19 0.07 4.52 0.20 0.18 0.00 0.00 0.07 0.09 0.00 0.00 0.03 0.02 0.00 0.00 0.00 Bacteria; Proteobacteria; Deltaproteobacteria 
996 466 0.95 0.09 0.02 0.00 0.63 0.84 4.68 0.34 0.45 0.10 0.06 0.07 0.11 0.19 0.16 0.18 0.05 0.00 0.04 0.00 Bacteria; Proteobacteria; Alphaproteobacteria; Sphingomonadales; Sphingomonadaceae; Sandarakinorhabdus 
42 417 0.48 0.00 0.00 0.00 0.67 0.28 1.68 1.95 0.85 0.05 0.11 0.05 0.11 0.02 0.10 0.36 0.02 0.07 0.19 0.01 Bacteria; Proteobacteria; Alphaproteobacteria; Sphingomonadales 
980 439 0.81 0.02 0.03 0.00 0.32 0.33 2.10 2.21 0.36 0.07 0.21 0.11 0.06 0.10 0.07 0.27 0.00 0.03 0.37 0.03 Bacteria; Proteobacteria; Alphaproteobacteria; Rhizobiales 
160 434 0.88 0.49 0.44 0.45 0.38 0.30 1.68 0.20 0.17 0.03 1.17 0.34 0.11 0.17 0.19 0.13 0.00 0.21 0.12 0.16 Bacteria; Proteobacteria; Alphaproteobacteria; Rickettsiales; Rickettsiaceae; Rickettsia 
986 380 2.17 0.58 0.03 0.00 0.42 0.23 2.32 0.29 0.32 0.03 0.02 0.04 0.13 0.00 0.07 0.18 0.00 0.03 0.00 0.14 Bacteria; Proteobacteria; Alphaproteobacteria; Sphingomonadales; Sphingomonadaceae; Sphingomonas; alpina 
1731 348 0.07 0.00 0.10 0.21 0.59 0.44 0.66 0.12 1.33 0.09 0.05 0.04 0.21 0.00 0.15 0.15 0.05 0.29 0.84 0.30 Bacteria; Proteobacteria; Alphaproteobacteria; Rhizobiales 
3195 378 0.08 0.00 0.00 0.00 1.03 0.45 0.59 0.31 0.85 0.12 0.02 0.04 1.15 0.00 0.14 0.28 0.04 0.30 0.90 0.09 Bacteria; Proteobacteria; Alphaproteobacteria; Sphingomonadales; Sphingomonadaceae; Sphingomonas; BAC151 
102 264 0.00 0.00 0.02 0.11 0.21 0.15 0.54 0.25 0.38 0.20 0.53 0.07 0.15 0.03 0.14 0.31 0.90 0.06 0.18 0.18 Bacteria; Proteobacteria; Deltaproteobacteria 
2945 384 0.00 0.09 0.17 0.02 0.11 1.87 0.76 0.31 0.34 0.58 0.33 0.13 0.04 0.19 0.73 0.01 0.16 0.00 0.25 0.06 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Comamonadaceae; Roseateles; Mitsuaria 
2787 226 0.00 0.00 0.00 0.00 0.11 0.74 0.73 0.39 1.01 0.09 0.06 0.09 0.08 0.05 0.00 0.19 0.00 0.00 0.07 0.01 Bacteria; Proteobacteria; Alphaproteobacteria; Rickettsiales 
249 1,013 0.22 2.57 3.90 4.20 2.38 0.03 0.00 0.78 2.00 0.07 0.11 0.13 0.49 0.08 0.05 0.09 0.04 0.12 0.25 0.26 Bacteria; Planctomycetes; Phycisphaerae; Phycisphaerales; Phycisphaeraceae 
3144 1,121 2.34 3.45 3.22 2.69 1.56 0.07 0.19 0.47 1.78 0.26 0.05 0.11 1.06 0.13 0.00 0.07 0.07 1.10 0.35 0.48 Bacteria; Cyanobacteria; empty_class; Chroococcales; empty_family; Microcystis; aeruginosa 
3054 1,578 5.59 4.56 2.58 3.43 3.34 0.70 0.02 2.66 3.27 0.05 0.06 0.05 0.25 0.02 0.00 0.09 0.11 0.15 0.02 0.06 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
3072 1,678 3.67 5.95 4.19 1.36 1.93 1.26 2.81 2.87 2.06 0.31 0.29 0.36 0.28 0.08 0.25 0.25 0.20 0.47 0.46 0.37 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
1495 1,312 3.83 4.11 1.27 1.38 1.81 0.91 1.23 3.64 1.58 0.38 0.12 0.13 0.55 0.29 0.08 0.12 0.09 0.50 0.16 0.30 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
3131 1,088 7.80 1.01 0.54 0.78 2.86 0.22 0.38 1.07 2.59 0.05 0.03 0.04 0.34 0.07 0.04 0.01 0.07 0.14 0.07 0.10 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
926 4.13 1.58 0.71 1.18 2.90 0.13 0.05 1.56 2.46 0.07 0.06 0.04 0.25 0.07 0.05 0.06 0.07 0.08 0.04 0.07 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
3143 608 2.17 3.83 0.39 1.25 0.95 0.09 0.05 0.23 1.23 0.09 0.02 0.04 0.36 0.08 0.04 0.01 0.00 0.06 0.07 0.03 Bacteria; Cyanobacteria; empty_class; Chroococcales; empty_family; Microcystis; aeruginosa 
3145 569 1.43 2.80 0.71 1.51 0.61 0.07 0.07 0.22 1.37 0.07 0.02 0.09 0.43 0.24 0.00 0.01 0.02 0.11 0.11 0.18 Bacteria; Cyanobacteria; empty_class; Chroococcales; empty_family; Microcystis 
1457 907 4.41 2.29 1.02 0.76 1.72 0.45 0.61 0.98 1.61 0.19 0.09 0.07 0.66 0.17 0.08 0.07 0.07 0.12 0.11 0.11 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
1456 721 2.46 3.08 0.59 0.69 1.07 0.57 0.17 1.05 1.20 0.20 0.12 0.05 0.38 0.10 0.04 0.07 0.05 0.21 0.16 0.18 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
3112 604 1.82 1.95 1.07 0.80 0.92 0.30 0.35 1.26 1.09 0.10 0.05 0.07 0.06 0.07 0.04 0.07 0.02 0.17 0.11 0.03 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
979 3,169 1.33 0.83 0.08 0.00 9.27 2.52 2.10 21.26 8.61 0.22 0.08 0.22 0.43 0.32 0.14 1.50 0.09 0.21 0.21 0.17 Bacteria; Proteobacteria; Alphaproteobacteria; Rhizobiales 
50 2,679 1.09 3.32 0.05 0.00 4.21 3.48 6.08 18.06 3.30 0.34 0.14 0.22 1.19 0.25 0.29 0.87 0.33 0.25 0.47 0.07 Bacteria; Proteobacteria; Alphaproteobacteria; Caulobacterales; Hyphomonadaceae 
1192 3,486 5.21 12.82 4.68 7.91 2.27 1.36 3.74 12.53 2.67 1.13 0.41 0.47 0.75 0.20 0.37 0.75 0.11 0.80 0.81 1.36 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
3022 3,703 1.74 6.08 21.43 14.16 2.78 0.44 0.00 2.27 2.68 0.15 0.33 0.72 2.15 0.49 1.20 0.40 0.20 0.28 0.93 5.06 Bacteria; Proteobacteria; Alphaproteobacteria 
721 3,239 8.08 2.55 8.26 8.51 4.37 0.33 0.00 0.92 5.05 0.44 0.65 1.01 3.83 1.03 1.83 0.94 0.11 0.70 1.42 4.14 Bacteria; Proteobacteria; Alphaproteobacteria 
3084 444 1.89 1.54 0.31 0.26 0.88 0.23 0.12 0.80 0.87 0.03 0.03 0.02 0.23 0.00 0.00 0.00 0.02 0.25 0.00 0.09 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
3113 381 2.41 1.05 0.31 0.32 0.48 0.10 0.17 0.72 0.49 0.07 0.02 0.02 0.15 0.03 0.01 0.06 0.00 0.07 0.02 0.03 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
1649 420 2.06 2.29 0.36 0.54 0.38 0.10 0.07 0.04 0.50 0.14 0.00 0.00 0.30 0.13 0.08 0.04 0.04 0.19 0.09 0.11 Bacteria; Proteobacteria; Alphaproteobacteria; Rhodospirillales; Acetobacteraceae; Roseomonas; NA; SK_65; NA 
1455 350 0.93 1.84 0.03 0.34 0.61 0.30 0.07 0.78 0.38 0.10 0.03 0.02 0.09 0.07 0.01 0.10 0.02 0.26 0.00 0.09 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
3098 343 1.54 1.84 0.15 0.19 0.61 0.07 0.02 0.42 0.46 0.15 0.00 0.00 0.28 0.00 0.01 0.00 0.02 0.12 0.09 0.07 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
3132 266 1.39 0.45 0.20 0.35 0.61 0.12 0.07 0.23 0.70 0.03 0.02 0.02 0.11 0.03 0.01 0.03 0.04 0.01 0.04 0.01 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
3056 228 0.66 0.92 0.12 0.11 0.46 0.16 0.07 0.35 0.70 0.02 0.00 0.04 0.19 0.05 0.00 0.01 0.00 0.03 0.02 0.00 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
3086 225 0.80 0.71 0.07 0.24 0.38 0.16 0.12 0.42 0.38 0.07 0.00 0.02 0.11 0.02 0.01 0.06 0.02 0.18 0.00 0.06 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
984 564 0.30 0.15 0.00 0.00 2.76 0.61 0.45 1.43 2.56 0.10 0.03 0.02 0.26 0.12 0.11 0.06 0.00 0.08 0.04 0.03 Bacteria; Proteobacteria; Alphaproteobacteria; Rhizobiales 
2338 913 0.35 0.30 0.02 0.02 6.23 0.29 0.45 0.04 5.76 0.12 0.03 0.04 0.87 0.00 0.41 0.00 0.04 0.03 0.07 0.00 Bacteria; Proteobacteria; Alphaproteobacteria; Sphingomonadales; Sphingomonadaceae; Novosphingobium; aromaticivorans; NA; NA 
983 880 0.10 0.00 0.00 0.00 3.79 0.55 4.26 0.25 3.75 0.15 0.15 0.13 1.45 0.07 0.32 0.04 0.04 0.25 0.33 0.00 Bacteria; Proteobacteria; Alphaproteobacteria; Sphingomonadales; Sphingomonadaceae; Sphingomonas 
139 533 2.12 0.00 0.02 0.02 2.67 0.01 0.00 0.00 2.46 0.00 0.00 0.00 1.26 0.10 0.00 0.00 0.04 0.14 0.00 0.01 Bacteria; Chlamydiae; Chlamydiae; Chlamydiales; Chlamydiaceae 
74 624 2.24 0.41 0.20 0.11 3.15 0.03 0.02 0.01 2.46 0.14 0.00 0.02 1.06 0.08 0.03 0.03 0.09 0.18 0.02 0.20 Bacteria; Chlamydiae; Chlamydiae; Chlamydiales 
2943 462 1.56 0.04 0.03 0.09 2.36 0.32 0.14 0.18 1.66 0.29 0.02 0.02 0.53 0.03 0.08 0.06 0.13 0.03 0.09 0.04 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Comamonadaceae; Pelomonas 
2944 348 1.92 0.00 0.00 0.00 2.48 0.17 0.07 0.16 0.60 0.02 0.00 0.00 0.17 0.02 0.01 0.03 0.04 0.07 0.21 0.00 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Comamonadaceae; Inhella 
340 221 0.43 0.00 0.00 0.00 0.95 0.06 0.02 0.04 1.29 0.02 0.00 0.00 0.60 0.02 0.00 0.03 0.11 0.01 0.02 0.01 Bacteria; Proteobacteria; Deltaproteobacteria; Bdellovibrionales; Bacteriovoracaceae; Peredibacter 
1013 245 0.03 0.69 0.00 0.00 2.31 0.01 0.05 0.00 0.98 0.00 0.02 0.00 0.23 0.00 0.01 0.01 0.00 0.00 0.00 0.03 Bacteria; Proteobacteria; Alphaproteobacteria; Sphingomonadales; Sphingomonadaceae; Sphingomonas 
3239 2,214 0.00 0.11 0.05 0.04 0.10 0.04 0.09 0.03 0.06 0.00 0.02 0.11 19.51 9.20 0.49 0.04 0.94 0.12 8.71 0.04 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Comamonadaceae; Pseudorhodoferax 
3242 1,398 0.00 0.04 0.07 0.15 0.00 0.03 0.07 0.00 0.00 0.00 0.20 0.00 0.49 14.95 0.43 0.03 0.00 0.28 6.71 0.26 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Comamonadaceae 
1689 5,588 0.00 0.11 0.00 0.00 0.00 0.06 0.21 0.03 0.08 9.26 1.82 1.25 3.77 6.89 15.90 29.28 0.74 3.51 4.44 8.09 Bacteria; Bacteroidetes; Sphingobacteriia; Sphingobacteriales; Chitinophagaceae; Lacibacter 
1389 3,245 0.07 0.13 0.00 0.00 0.10 0.04 0.05 0.04 0.34 3.10 3.45 2.94 8.53 4.68 0.78 3.00 0.60 8.39 8.19 7.55 Bacteria; Bacteroidetes; Sphingobacteriia; Sphingobacteriales; Chitinophagaceae; Sediminibacterium 
45 2,210 0.00 0.00 0.00 0.00 0.02 0.00 0.00 0.00 0.32 2.40 3.12 1.95 3.74 4.90 2.45 2.50 2.80 3.54 7.34 0.97 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Oxalobacteraceae; Paucimonas 
3206 3,202 0.48 0.11 0.02 0.00 0.06 0.03 0.05 0.07 0.52 7.59 2.55 2.29 10.24 5.76 4.92 4.84 0.85 2.80 7.32 2.06 Bacteria; Proteobacteria; Alphaproteobacteria; Sphingomonadales; Sphingomonadaceae; Novosphingobium; FW − 6 
2570 1,585 0.05 0.00 0.02 0.02 0.11 0.00 0.02 0.00 0.08 0.05 0.17 0.00 0.19 3.35 8.56 7.57 0.00 0.14 2.81 0.65 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Comamonadaceae; Pseudorhodoferax 
128 1,137 0.00 0.00 0.00 0.00 0.00 0.04 0.00 0.00 0.04 0.12 0.05 0.04 0.98 1.69 4.92 3.61 0.05 3.79 1.46 0.10 Bacteria; Proteobacteria; Deltaproteobacteria 
1852 1,066 0.00 0.00 0.00 0.00 0.00 0.07 0.00 0.00 1.94 0.14 0.29 0.00 0.15 2.02 5.22 3.48 0.00 0.70 1.84 0.03 Bacteria; Proteobacteria; Alphaproteobacteria; Caulobacterales; Caulobacteraceae; Caulobacter; Brevundimonas 
269 956 0.00 0.00 0.02 0.04 0.02 0.00 0.02 0.00 0.04 0.03 0.00 0.00 3.58 4.50 2.35 0.84 0.20 0.22 3.86 0.21 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Comamonadaceae; Polaromonas 
123 597 0.00 0.00 0.03 0.00 0.13 0.59 0.09 0.00 0.07 0.00 0.00 0.02 1.09 2.65 2.80 1.15 0.02 0.06 0.58 0.04 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Comamonadaceae; NA; Piscinibacter 
924 668 0.05 0.21 0.00 0.00 0.00 0.01 0.09 0.00 0.10 0.03 0.11 0.02 0.45 1.58 2.75 1.20 0.02 0.55 1.55 1.51 Bacteria; Proteobacteria; Betaproteobacteria 
1650 434 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.01 0.00 6.15 Bacteria; Proteobacteria; Alphaproteobacteria; Rhodospirillales; Acetobacteraceae; Roseomonas; gilardii; NA; NA; subsp._rosea 
146 998 0.00 0.00 0.00 0.00 0.06 0.00 0.00 0.00 0.07 0.68 0.64 0.42 0.02 1.77 0.16 0.82 0.11 0.12 7.03 4.22 Bacteria; Verrucomicrobia 
145 643 0.12 0.00 0.07 0.00 0.06 0.09 0.09 0.06 0.13 0.03 0.24 0.18 0.06 0.13 0.99 1.75 0.23 0.18 3.58 2.10 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae; Planctomyces 
2148 351 0.00 0.00 0.02 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.02 0.07 0.99 0.28 0.00 0.32 3.90 0.13 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae; Planctomyces 
2573 344 0.00 0.02 0.02 0.04 0.11 0.62 0.26 0.00 0.10 0.02 0.23 0.02 0.98 1.40 0.82 0.01 0.16 0.06 0.79 0.03 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Comamonadaceae; Acidovorax 
1994 428 0.00 0.00 0.00 0.00 0.04 0.00 0.00 0.00 0.06 1.47 0.14 0.40 0.26 0.86 1.51 0.31 0.00 0.29 1.26 0.23 Bacteria; Proteobacteria; Alphaproteobacteria; Rhodospirillales; Acetobacteraceae; Gluconacetobacter; diazotrophicus; ICB557; NA 
1151 296 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.34 0.66 1.11 1.53 0.00 0.02 0.00 1.12 0.00 Bacteria; Cyanobacteria; Chloroplast 
1849 270 0.02 0.00 0.00 0.00 0.00 0.01 0.00 0.00 0.00 0.00 0.00 0.00 1.28 0.71 1.55 0.21 0.00 0.00 0.54 0.00 Bacteria; Proteobacteria; Alphaproteobacteria; Rhodobacterales; Rhodobacteraceae; Rhodobacter 
1734 337 0.00 0.00 0.00 0.00 0.02 0.07 0.00 0.03 0.11 0.46 0.20 0.09 1.15 0.37 0.40 1.83 0.13 0.14 0.46 0.03 Bacteria; Proteobacteria; Alphaproteobacteria; Caulobacterales; Caulobacteraceae; Brevundimonas 
680 328 0.00 0.04 0.02 0.00 0.00 0.00 0.02 0.00 0.22 0.14 0.03 0.00 1.81 0.15 0.25 1.33 0.54 0.19 0.12 0.50 Bacteria; Proteobacteria; Gammaproteobacteria; Xanthomonadales; Xanthomonadaceae; Thermomonas; R039N 
2685 228 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.06 0.03 0.44 0.11 0.00 0.54 0.12 0.27 0.02 0.14 1.98 0.06 Bacteria; Bacteroidetes; Sphingobacteriia; Sphingobacteriales 
1265 206 0.00 0.00 0.00 0.06 0.06 0.03 0.00 0.01 0.17 0.09 0.05 0.00 0.15 0.44 0.43 0.25 0.00 0.61 0.76 0.11 Bacteria; Proteobacteria; Alphaproteobacteria; Caulobacterales; Hyphomonadaceae; Hirschia 
1956 195 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.56 0.06 0.27 0.09 0.20 1.04 0.24 0.00 0.26 0.07 0.16 Bacteria; Proteobacteria; Alphaproteobacteria; Rhizobiales; Hyphomicrobiaceae; Hyphomicrobium 
3223 194 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.32 0.26 0.05 0.08 0.22 0.43 0.00 0.02 1.35 0.00 0.11 Bacteria; Proteobacteria; Alphaproteobacteria; Rickettsiales 
2470 262 0.00 0.00 0.00 0.00 0.06 0.00 0.14 0.04 0.39 0.03 0.08 1.01 0.04 0.00 0.27 0.07 0.54 1.24 0.21 0.00 Bacteria; Proteobacteria; Betaproteobacteria; Nitrosomonadales; Nitrosomonadaceae 
14 347 0.00 0.00 0.00 0.00 0.00 0.00 0.02 0.01 0.00 0.39 0.43 0.99 0.13 0.13 0.60 0.34 0.07 0.61 0.07 1.51 Bacteria; Acidobacteria 
621 499 0.00 0.00 0.03 0.07 0.00 0.00 0.02 0.00 0.03 0.73 1.38 1.08 0.00 0.39 0.65 0.60 0.23 0.48 0.11 1.88 Bacteria; Proteobacteria; Betaproteobacteria; Nitrosomonadales; Nitrosomonadaceae 
76 536 0.00 0.02 0.00 0.02 0.00 0.00 0.00 0.01 0.36 0.70 0.73 0.87 0.13 0.37 0.48 0.49 0.51 1.50 1.33 0.85 Bacteria; Proteobacteria; Deltaproteobacteria; Myxococcales 
182 426 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.01 0.48 0.90 2.17 0.02 0.32 0.26 0.85 0.67 0.21 0.37 0.70 Bacteria; Verrucomicrobia 
2778 291 0.00 0.00 0.00 0.00 0.10 0.00 0.07 0.04 0.11 0.14 0.55 0.88 0.02 0.19 0.37 0.45 0.49 0.40 0.18 0.63 Bacteria; Proteobacteria; Alphaproteobacteria; Rhodospirillales; Rhodospirillaceae 
1961 955 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.15 2.24 0.98 0.00 2.14 0.55 0.16 0.13 4.51 0.14 3.20 Bacteria; Proteobacteria; Alphaproteobacteria; Rhodobacterales; Rhodobacteraceae; Pseudorhodobacter; Rhodobacter; NA; 16 − 62; NA 
2683 904 0.00 0.00 0.03 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.03 1.32 0.11 0.59 0.80 0.70 0.00 3.76 0.00 5.80 Bacteria; Bacteroidetes; Sphingobacteriia; Sphingobacteriales 
285 538 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.06 0.29 0.00 0.78 0.07 0.49 0.00 3.38 0.44 2.33 Bacteria; Proteobacteria; Alphaproteobacteria; Sphingomonadales; Sphingomonadaceae; Zymomonas 
15 678 0.00 0.00 0.15 0.09 0.04 0.00 0.00 0.04 0.00 0.00 0.05 0.20 0.00 0.22 1.54 2.64 0.00 2.98 0.04 1.79 Bacteria; Proteobacteria; Betaproteobacteria; Methylophilales; Methylophilaceae 
1957 583 0.03 0.04 0.00 0.02 0.00 0.00 0.00 0.04 0.01 0.92 0.47 0.47 0.06 0.39 1.13 1.23 0.20 1.89 0.12 1.68 Bacteria; Proteobacteria; Alphaproteobacteria; Rhodospirillales; empty_family; Reyranella 
2334 772 0.00 0.00 0.02 0.00 0.00 0.00 0.00 0.00 0.01 1.26 0.67 1.54 0.17 0.54 1.98 0.55 0.00 2.76 0.07 2.00 Bacteria; Proteobacteria; Alphaproteobacteria; Rickettsiales 
3219 714 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.01 1.09 0.64 1.44 0.08 0.35 1.03 0.78 0.02 3.03 0.11 2.10 Bacteria; Proteobacteria; Alphaproteobacteria; Rickettsiales 
110 1,531 0.00 0.00 0.02 0.02 0.17 0.00 0.05 0.04 0.01 0.97 1.89 4.55 0.02 0.08 0.04 0.19 18.16 0.47 0.04 0.26 Bacteria; Verrucomicrobia; Opitutae; Opitutales; Opitutaceae; Opitutus 
1242 1,440 0.00 0.00 0.00 0.00 0.04 0.01 0.00 0.00 0.06 0.00 0.00 0.00 0.06 0.00 0.00 0.01 25.73 0.00 0.09 0.00 Bacteria; Chlamydiae; Chlamydiae; Chlamydiales; Simkaniaceae; Candidatus_Rhabdochlamydia 
94 612 0.00 0.00 0.00 0.00 0.02 0.01 0.09 0.01 0.24 0.44 0.99 1.08 0.02 0.13 0.45 0.33 3.96 1.46 0.07 0.63 Bacteria; Gemmatimonadetes; Gemmatimonadetes; Gemmatimonadales; Gemmatimonadaceae 
2145 603 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.01 0.00 0.44 1.28 0.56 0.00 0.00 0.00 0.16 5.62 1.34 0.04 0.57 Bacteria; Proteobacteria; Alphaproteobacteria; Rhodospirillales 
16 305 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.03 0.00 0.56 1.05 1.14 0.00 0.00 0.00 0.00 2.48 0.01 0.00 0.00 Bacteria; Nitrospirae; Nitrospira 
180 250 0.00 0.00 0.00 0.00 0.00 0.00 0.05 0.01 0.01 0.12 1.05 0.00 0.00 0.02 0.03 0.01 2.98 0.00 0.02 0.00 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae; Planctomyces 
2895 716 0.00 0.00 0.02 0.00 0.00 0.00 0.00 0.03 0.21 0.00 0.00 0.02 0.02 0.94 0.22 5.76 3.99 0.21 0.00 0.06 Bacteria; Verrucomicrobia 
32 435 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 7.86 0.00 0.00 0.00 Bacteria; Verrucomicrobia 
82 304 0.00 0.02 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.02 0.02 0.02 0.00 0.00 0.00 0.00 5.42 0.00 0.00 0.00 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae; Gemmata 
2896 505 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.11 0.02 0.11 6.39 0.04 0.00 0.19 0.12 0.22 0.04 0.14 1.25 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae; Planctomyces 
124 370 0.00 0.00 0.00 0.00 0.00 0.00 0.02 0.01 0.00 0.36 1.14 3.67 0.02 0.00 0.00 0.00 0.47 0.54 0.00 0.04 Bacteria; Proteobacteria; Betaproteobacteria; Nitrosomonadales; Nitrosomonadaceae 
1733 291 0.00 0.06 0.00 0.00 0.04 0.01 0.17 0.57 0.06 0.19 0.88 2.67 0.04 0.00 0.01 0.04 0.14 0.01 0.02 0.03 Bacteria; Proteobacteria; Alphaproteobacteria; Rhodospirillales; Rhodospirillaceae 
126 224 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.01 0.56 0.82 1.14 0.00 0.00 0.15 0.04 0.38 0.50 0.00 0.04 Bacteria; Acidobacteria 
65 360 0.03 0.02 0.02 0.06 0.00 0.00 0.05 0.01 0.06 0.87 1.58 2.02 0.02 0.00 0.15 0.04 0.23 0.08 0.07 0.58 Bacteria; Planctomycetes; Planctomycetacia; Planctomycetales; Planctomycetaceae 
113 216 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.63 1.49 1.46 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 Bacteria 
97 255 0.00 0.00 0.00 0.02 0.04 0.03 0.02 0.04 0.01 0.77 1.51 1.32 0.00 0.00 0.03 0.01 0.36 0.04 0.04 0.00 Bacteria; Proteobacteria; Alphaproteobacteria; Rhodospirillales 
620 410 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 1.77 2.27 1.79 0.00 0.02 0.01 0.01 0.04 0.73 0.00 0.00 Bacteria; Proteobacteria; Betaproteobacteria; Nitrosomonadales; Nitrosomonadaceae 
3196 572 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.03 2.05 3.22 2.33 0.02 0.07 0.10 0.00 1.66 0.03 0.00 0.04 Bacteria; Proteobacteria; Alphaproteobacteria; Rickettsiales 
1735 418 0.00 0.00 0.00 0.00 0.00 0.01 0.05 0.00 0.03 2.17 1.40 2.62 0.00 0.00 0.00 0.00 0.47 0.23 0.04 0.06 Bacteria; Proteobacteria; Alphaproteobacteria; Rhodospirillales; empty_family; Reyranella 
3186 521 0.00 0.02 0.00 0.00 0.00 0.00 0.00 0.00 0.20 1.82 0.70 1.34 0.15 0.12 1.61 0.16 0.23 1.52 0.04 0.16 Bacteria; Planctomycetes; Phycisphaerae; Phycisphaerales; Phycisphaeraceae 
1149 287 0.00 0.00 0.02 0.07 0.00 0.01 0.02 0.06 0.41 1.31 0.21 2.15 0.00 0.05 0.01 0.16 0.29 0.00 0.05 0.04 Bacteria 
2146 522 0.00 0.00 0.00 0.00 0.13 0.00 0.00 0.04 0.22 6.39 0.49 0.27 0.21 0.00 0.01 0.03 0.45 0.47 0.02 0.00 Bacteria; Proteobacteria; Alphaproteobacteria; Caulobacterales; Caulobacteraceae; Caulobacter; Es3; MDB1–11; NA 
24 315 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 5.34 0.00 0.00 0.00 0.00 0.00 0.00 0.04 0.00 0.00 0.00 Bacteria; Verrucomicrobia 
261 290 0.10 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 3.07 0.04 0.02 0.00 0.00 0.00 0.00 1.09 0.00 0.00 Bacteria 
311 470 0.02 0.00 0.00 0.00 0.11 0.04 0.33 0.07 0.80 0.26 4.23 1.28 0.02 0.02 0.01 0.04 0.07 0.03 0.09 0.04 Bacteria; Proteobacteria; Deltaproteobacteria 
163 755 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.02 5.81 2.82 0.00 0.00 0.00 0.00 0.00 2.95 0.00 0.03 Bacteria; Bacteroidetes; Sphingobacteriia; Sphingobacteriales 
2231 845 0.02 0.00 0.00 0.19 0.02 0.03 0.00 0.01 0.01 0.80 9.35 2.00 0.00 0.00 0.00 0.22 0.05 0.45 0.02 0.06 Bacteria; Proteobacteria 
1386 2,909 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 20.34 10.55 12.66 0.00 0.03 0.04 0.01 0.07 4.05 0.00 0.28 Bacteria; Bacteroidetes; Sphingobacteriia; Sphingobacteriales; Chitinophagaceae; Ferruginibacter 
925 1,808 0.00 0.00 0.02 0.02 0.00 0.00 0.00 0.00 0.06 3.26 7.01 5.24 0.02 0.07 2.35 3.33 2.42 2.98 0.09 1.52 Bacteria; Proteobacteria; Betaproteobacteria 
2143 1,324 0.10 0.02 0.12 0.28 0.21 0.04 0.07 0.07 0.45 1.36 2.54 2.35 0.11 1.31 1.84 1.36 0.78 5.64 0.97 0.70 Bacteria; Proteobacteria; Alphaproteobacteria; Rhizobiales 
3184 1,131 0.00 0.06 0.03 0.02 0.02 0.00 0.00 0.03 0.55 2.63 1.72 3.88 0.43 0.25 2.76 1.29 0.98 2.49 0.07 0.53 Bacteria; Planctomycetes; Phycisphaerae; Phycisphaerales; Phycisphaeraceae 

To be included in Figure 1 and this table, a given ASV must have at least a 1.0% presence in one of the collection sites for at least one sampled month. Each ASV is assigned to microbial assemblage 1 or 2 or 3 or 4 based upon where they were found as determined by the MED analysis. Each sample was found to contain multiple ASVs drawn from different bacterial phyla, and most phyla are represented in all samples. However, the particular ASV identified is quite different from the different assemblages found in the different sampling sites and months.

Sample collection and filtration

Prior to sampling, water was allowed to flow from each tap for about 3 min, followed by flame sterilization of the faucet. We collected 2.5 L of water samples over a 3-min time interval into pre-sterilized bottles. The samples were maintained at 4 °C and processed within 4 h of collection. Temperature and pH were determined at the time of sampling. Microorganisms were harvested by filtering 1.9 L of each water sample through a 0.2 μm Sterivex™ GP filter unit (Millipore, Billerica, MA) and stored at −18 °C until DNA isolation.

Nucleic acid extraction and purification

Nucleic acids were extracted from the Sterivex™ GP filter units using the Puregene DNA extraction kit (Gentra Systems, Minneapolis, MN) following the manufacturer's protocol with modifications (Sinigalliano et al. 2007). The reservoir of each filter cartridge was flooded with 2 mL of Puregene lysis buffer and 10 μL of the lytic enzyme (Qiagen, Valencia, CA) and incubated at 37 °C for 30 min with occasional shaking. Ten microlitres of Proteinase K (Qiagen, Valencia, CA) was added, and the sample was mixed thoroughly and incubated at 60 °C for 70 min with occasional agitation. The lysed cell suspensions were removed from the reservoir surrounding the filters using sterile 3 mL syringes and transferred into 2 mL sterile screw-cap tubes. Two hundred and thirty-five microlitres of protein precipitation solution (Qiagen, Valencia, CA) was then added, and the suspensions were incubated on ice for 15 min followed by centrifugation at 12,000 × g for 5 min. The supernatants were transferred into tubes containing 930 μL of 100% isopropanol. DNAs were pelleted by centrifugation at 12,000 × g for 5 min, washed with 700 μL of ice cold 70% ethanol, and the suspensions were centrifuged again at 12,000 × g for 5 min. The DNA pellets were air-dried at room temperature and resuspended in 30 μL of DNA hydration buffer (Qiagen, Valencia, CA). The quality of the extracted DNAs was assessed by agarose gel electrophoresis, and concentrations were determined on a Nanodrop ND-1000 (Nanodrop Technologies, Wilmington, DE).

Polymerase chain reaction

PCR amplification of the V4 through V6 hypervariable regions of bacterial 16S rRNA genes employed fusion primers as described previously (Newton et al. 2013). We generated PCR amplicons in triplicate in 30 μL reaction volumes and included a separate no-template negative control. Reactions were performed in a GeneAmp PCR System 2720 cycler (Applied Biosystems, Foster City, CA) with amplification conditions described previously (Newton et al. 2013).

The resulting PCR products were purified using AMPure size exclusion beads (Agencourt Bioscience Corporation, Beverly, MA) to eliminate fragments below 400 bp and eluted in 24 μL of Buffer EB. The quality and size distribution of the amplicons were assessed on a Bioanalyzer 2100 (Agilent, Palo Alto, CA) using a DNA 1000 Lab Chip™ while the concentrations were determined using the Invitrogen Quant-IT™ Picogreen assay (Molecular Probes, Carlsbad, CA) with fluorescence measured on a TBS 380 Minifluorometer (Turner Biosystems, Promega, Madison, WI). PCR products with unique barcode sequences were pooled together at equimolar concentrations (25 ng each) for emulsion PCR.

Sequencing

We amplified the pooled DNA products in PCR mixture-in-oil emulsion and sequenced from the V6 end of the amplicon using Roche GS-FLX Titanium (Roche, Switzerland) 454 protocols.

Sequence analysis

Our bioinformatics process removed chimeras and low-quality reads as previously described (Huse et al. 2007). Reads were mapped to a bacterial reference alignment, degapped and re-trimmed to an average length of 261 nt (V5–V6) using PyNAST (Caporaso et al. 2010) and oligotyping tools (Eren et al. 2013). The final aligned dataset served as input to the Minimum Entropy Decomposition (MED) pipeline, an information theory-based clustering algorithm for sensitive partitioning of high-throughput marker gene sequences followed by the determination of their co-occurrence in various environments (Eren et al. 2015). MED identifies ‘amplicon sequence variants’ (ASVs), previously called MED nodes or oligotypes (Callahan et al. 2017). ASVs were assigned rank-based taxonomy using vsearch v1.9.5 (Rognes et al. 2016) and the silva v119 reference database (Quast et al. 2013).

MED pipeline-linked analyses

First, each sample (location and month) was analyzed with regard to the detected ASVs and their relative abundances. Second, the samples were arranged according to whether they shared ASVs, and the degree of similarity is plotted on the horizontal distribution (see Figure 1). Third, the detailed analysis of the abundance of all 154 ASVs with the presence of at least 1% in at least one sample month is presented in Table 1 and in the vertical distribution of Figure 1. Note that additional 178 ASVs were detected whose abundance was below the 1% cut-off. The entire list of detected ASVs is presented in Supplementary Table 1. Of note, some different ASVs with the same family name are located at different positions in the taxonomic list. This is because the different individual members are represented by different, but related 16S RNA sequences and presumably have different genome constituents, and thus different and perhaps unique physiological capabilities.

RESULTS AND DISCUSSION

Results

General water quality metrics

pH ranged between 6.6 and 7.6, within WHO standards for drinking water (6.5–8.5). Temperature variation ranged between 27 and 33.5 °C. There were no significant differences in either pH or temperature between sites on the same sampling date, but there were significant differences in both metrics between different months (e.g. November pH = 7.00 vs. August pH = 7.48).

Bacterial diversity

We recovered an average of 11,839 and 14,159 sequences from the September 2009 through December 2009 waterworks and tap samples. Water samples from polytanks A, B and C yielded averages of 14,466, 12,572 and 12,247 sequences, respectively.

Figure 1 presents a heat map that displays the relative abundance of 154 ASVs versus their occurrence in different sample sources (each sample site is color-coded in both Figure 1 and Supplementary Figure 1). Table 1 lists these ASVs in the same order as Figure 1 along with their percentage relative abundance and taxonomic affinity. Inclusion of an ASV in Table 1 and Figure 1 required their relative abundance to be equal or exceeded 1.0% of the reads in one of the collection sites for at least 1 month. The relative abundance of any given ASV in any of the 20 samples is indicated by the color of the sample bar [red indicates high abundance (>16% representation in the sample), blue for low abundance (<1% or not observed)]. This beta diversity analysis identified four MED assemblages. With the exception of the September sample from polytank A, assemblages 1 and 2 dominated the waterworks and tap samples, and assemblages 3 or 4 dominated polytank samples. The enumeration of major taxa for each site also identified the same difference patterns between sampling sites and months (Table 1). We observed distinctive community composition for the different sites despite their having a common source, the Weija waterworks. All samples included representatives of most phyla, but each sample contained multiple ASVs that resolved to taxa that did not appear in other samples. Several distinct MED nodes resolved to the same taxonomic assignments. For all of the sampling sites (except for the polytank A September sample), the identified taxa fell into the same assemblages from month to month, although there were considerable differences in relative abundances of taxa in different months.

Analysis of the abundant microbial taxa (genus-level classification) at the waterworks showed the presence of fairly constant qualitative genus distributions with considerable quantitative changes each month. Household sample populations also showed constant qualitative genus distributions with considerable quantitative changes each month. However, comparative analysis revealed that, with the exception of September, the dominant microbial community at the waterworks and tap varied drastically from the dominant bacterial populations observed in all but one of the polytank samples (Figure 1). These results demonstrate the presence of distinctive bacterial populations at the various sampling points.

Discussion

Our analysis described the diversity and relative taxon abundance for bacterial communities in a metropolitan DWDS in western Accra, Ghana. Sequence comparisons with existing databases revealed wide bacterial diversity with differing membership at each sampling point. There were members unique to each of the samples, and in general, the different sources manifested distinct populations with month-to-month changes in their relative abundance. For instance, the waterworks samples for all months were dominated by taxa from cluster 1 (Figure 1), but with different subpopulations in different months, while the household tap samples were dominated by taxa from cluster 2. This suggests that although the waterworks and household tap samples are related, there is likely some contamination and/or selection for different water taxa subsets during the delivery of water to the household taps. It would be of interest to study the relationship of microbial populations in other tap sources. The differences between most polytank samples (all but one, September polytank A) and the waterworks and household tap samples were dramatic for all sampling periods. Most polytank samples were dominated by taxa from clusters 3 or 4.

The origins of dominant taxa in the tap samples and polytank samples are not known definitively by the data in Table 1. However, we can look at whether the dominant taxa in the tap samples are present at detectable levels in the relevant waterworks samples (and thus could be potentially seeded by waterworks water), and we could do a similar analysis of the dominant polytank taxa (Table 1). For instance, the tap sample from December was dominated (21.26%) by ASV #979 that was found at low abundance in the October (0.83%) and November (0.08%) waterworks samples (Table 1). Thus, the ASV #979 could have been seeded by waterworks water. On the other hand, the dominant taxon in polytank A samples (ASV #1386 for October (20.34%), November (10.55%) and December (12.66%)) was not detectable in either the tap or the waterworks samples (Table 1), suggesting that contamination after the waterworks treatment contributed to the presence of this taxon in the polytanks. Regardless of the original source of this and other taxa in the tap and polytank sources, it is likely that the high abundance of various taxa reflects selection pressures. It should be noted that the polytanks offered distinct habitats compared to the waterworks and taps.

The September polytank A taxa types were unique in that they primarily fell into clusters 1 and 2 similar to the September tap and waterworks samples (Figure 1). A possible explanation for this result is that polytank A was cleaned (emptied, cleaned with household detergent and rinsed) a few days before the September sampling occurred, so it may not have had time to undergo the contamination/selective community change experienced by polytanks B and C. By October, this distinction had disappeared. It should be noted that these observations are consistent with the observation that water stagnation can give rise to distinct microbial assemblages in water taps (Ling et al. 2018).

It is of interest to determine whether any of the taxa found in the water system were likely to be human pathogens. It is not possible to definitively determine this without complete genome sequences and/or experimental tests. Nonetheless, the MED analysis identified taxa from some orders worthy of further investigation. Of particular concern were taxa from the Burkholderiales that were represented by several ASVs found in several sites including samples from the waterworks, tap source and various polytanks. Pathogenic strains resistant to antibiotics and disinfectants have been identified within the Burkholderiales. Waterborne disease outbreaks associated with this order have been reported previously (Inglis et al. 1999). Another order of particular concern is the Rickettsiales that were represented by several taxa mostly restricted to tap water and polytank sources. Rickettsiales are typically obligate intracellular pathogens of free-living amoeba in drinking water ecosystems (Fritsche et al. 1999).

The results from these studies indicate that while the water quality of the treated waterworks samples may be similar to that of other water plants (Eichler et al. 2006), the water quality for the ultimate consumers also depends upon the delivery system and storage systems. Storage in polytanks results in unique microbial contamination problems that should be considered in their design and/or maintenance regardless of efforts to improve the quality of water at the waterworks.

CONCLUSIONS

  • 1.

    Water treatment plants can provide water contaminated with complex microbial assemblages that are similar month to month over a 4-month period.

  • 2.

    Water taps provide water with microbial assemblages similar to that found in the water plant water.

  • 3.

    Water storage tanks newly cleaned and filled with water have a microbial assemblage similar to that observed in the tap water.

  • 4.

    Water stored for a month or more contains microbial assemblages distinct from those found in the water plant, in the tap and in the newly filled water tank.

  • 5.

    The existence of distinct microbial assemblages in stored water suggests that the provision of adequate dependable quantities of water (thus bypassing the need for domestic water storage) should be a priority in water system improvements or alternatively that water storage tanks should be cleaned at frequent intervals.

FUNDING

No funding source played a role in study design, collection or analysis and interpretation of data, nor did they play a role in manuscript writing or submission. The Alfred P. Sloan Foundation provides support for MBL personnel, supplies and equipment. This support played an important role in data analysis and data collection activities. The Wisconsin Alumni Research Foundation also supported activities related to collection and analysis through its funding of sample purification and sample data collection. The American Society for Microbiology International Professorships Program provided support for W.S. Reznikoff's participation as a 2009 Visiting Professor in the Department of Biochemistry, Cell and Molecular Biology at the University of Ghana. The Promega Corporation supported Dr G. Ecklu-Mensah's attendance and participation at the American Society for Microbiology 2011 annual meeting.

ACKNOWLEDGEMENTS

We thank the Ghana water company and all households for their permission and collaboration during water sampling and collection.

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