Well Water Sample Microbial Analysis

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1 Report Well Sample Microbial Analysis Submitted to XXX 23 November 2015 Submitted by Trevor Ghylin, P.E, Ph.D. and Tony Dallmier, Ph.D.

2 Summary Bacteria typically associated with human waste such as Bacteroides, Lachnospiracaea, Prevotella, E. coli and others were not found indicating that significant contamination with human waste is unlikely. Coliforms were also not detected, however Pseudomonas was detected, which is known to trigger a positive coliform result even though it is not a coliform. Though significant human waste contamination is not apparent, the presence of Bifidobacteria indicate potentially minor contamination with a fecal source as these are anaerobic bacteria typically found in mammal digestive systems. A small amount of potentially pathogenic genera were detected. These include Borrelia, Leptospira, Legionella, and Clostridium. Also, some problematic microorganisms including iron oxidizing and sulfate reducing bacteria which can corrode metal surfaces, discolor water, and produce foul odors in a well were found. Bacteria indicative of surface water intrusion (i.e. Limnohabitans) into the well were measured at high levels. Intrusion of surface water has the potential to contaminate wells with animal sources of fecal pollution and other contaminants. Also, some common slime forming bacteria such as Pseudomonas were found. Though evidence of significant contamination with human wastewater is not apparent, we would still recommend some disinfection or other treatment of this water prior to consumption as some problematic bacteria were found as well as evidence of surface water intrusion. Background One sample was received by Microbe Detectives on 9 October 2015 labelled Supply. Results DNA results are displayed in the DNAmy tables below. The results in these tables are a subset of the total data package focusing on bacteria that are typically important in drinking water systems. The full results are also provided in a spreadsheet. Results are presented as a percentage of total bacterial DNA in each sample. Limitations It should be noted that the presence of DNA does not mean the bacteria are either viable or pathogenic. The presence of DNA indicates the presence of a recently live or dead organism. The existence of an oxidizing compound such as chlorine will degrade DNA relatively rapidly.

3 Presently, high throughput DNA sequencing technology cannot distinguish at some species or a strain level. The taxonomic resolution is currently limited to genus level information. For example, the DNA sequence can show the presence of Campylobacter, but cannot determine whether the Campylobacter are viable or pathogenic, Campylobacter jejuni, or a related non-pathogenic species. The technology is continually improving and this limitation will likely diminish in the near future.

4 Table 1. DNAmy Summary of Bacteria Present (1-few, 10-abundant) Huntoon Pond Notes Coliforms 0 None detected Non-coliforms that trigger coliform test 2 Pseudomonas Fecal Indicators 0 None detected Contamination Indicators 2 Clostridia, Spirochaetae and Bifidobacterium are anaerobic bacteria; found in mammalian GI tracts; Borrelia can cause Lyme disease (needs zoonotic vector) or relapsing fever; Leptospira which can cause leptospirosis Potential Pathogens 2 Legionella and Clostridium found at very low levels Surface Bacteria 6 Potential surface water intrusion Iron Oxidation 10 High levels of iron oxidizing bacteria which can cause discoloration, corrosion and odors Sulfate Reduction 5 Can corrode metals and produce foul odors Sulfur Oxidation 2 Abundant and/or problematic bacteria are highlighted in red

5 Table 2. Phylum Distribution

6 Table 3. Genus Distribution

7 Copyright Microbe Detectives All Rights Reserved DNAmy (Microbe Detectives ) Bacterial Relative Abundance (% of Total Bacterial DNA) Bacteria Name Coliforms Pantoea 0.00% Escherichia 0.00% Enterobacter 0.00% Serratia 0.00% Citrobacter 0.00% Klebsiella 0.00% Hafnia 0.00% Proteus 0.00% Kluyvera 0.00% Buttiauxella 0.00% Erwinia 0.00% Hafvia 0.00% Obesumbacterium 0.00% Morganella 0.00% Proteus 0.00% Providencia 0.00% Rahnella 0.00% Raoultella 0.00% Trabusiella 0.00% Yersinia 0.00%

8 Non-Coliforms that can trigger Coliform test Streptococcus 0.00% Acinetobacter 0.00% Aeromonas 0.00% Bacillus 0.00% Chryseobacterium 0.00% Herbaspirillum 0.00% Microbacterium 0.00% Plesiomonas 0.00% Pseudomonas 0.27% Shewanella 0.00% Staphylococcus 0.00% Stenotrophomonas 0.00% Vibrio 0.00% Wautersia 0.00% Fecal Indicators Bacteroides 0.00% Prevotella 0.00% Escherichia 0.00% Ruminococcaceae 0.00% Enterococcus 0.00% Lachnospiraceae 0.00% Leptospira 0.00%

9 Contamination Indicators Enterobacteriales (Order) 0.00% Clostridia (Class, strictly anaerobic) 0.43% Spirochaetae (Phylum) 0.16% Fusobacterium 0.00% Streptococcus 0.00% Staphylococcus 0.00% Leptotrichia 0.00% Rothia 0.00% Bifidobacterium 0.18% Aggregatibacter 0.00% Trichococcus 0.00% Gemella 0.00% Porphyromonas 0.00% Capnocytophaga 0.00% Arcobacter 0.00% Potential Pathogens Mycobacterium 0.00% Helicobacter 0.00% Haemophilus 0.00% Legionella % Escherichia, Shigella 0.00% Salmonella 0.00% Campylobacter 0.00% Aeromonas 0.00% Alternaria 0.00% Clostridium 0.04% Freshwater or Marine Bacteria (potential indicators of surface water intrusion) Polynucleobacter 0.00% Prochlorococcus 0.05% Limnohabitans 3.83% Pelagibacter 0.00% aci Freshwater Actinobacteria 0.00%

10 Nitrogen Fixation Azospirillum 0.00% Acidithiobacillus 0.00% Rhizobiales (Order) 0.00% Desulfovibrio 0.41% Azospira 0.00% Bradyrhizobium 0.00% Methylosoma 0.00% Clostridium 0.04% Carbon Fixation Sediminibacterium 0.00% Sideroxydans 0.00% Nitrospira 0.00% Acidithiobacillus 0.00% Sulfuricurvum 0.12% Jettenia 0.00% Nitrosomonas 0.00% Sulfuricella 0.00% Geobacter 6.88% Gallionella 4.21% Brocadia 0.00% Scalindua 0.00% Leptospirillum 0.00% Crenothrix 7.00% Nitrospinaceae 0.00% Ferritrophicum 0.00% Candidatus, Nitrosoarchaeum 0.00% Nitrosococcus 0.00% Nitrosopumilus 0.00% Sulfurivirga 0.00%

11 Ammonia Oxidation Nitrosospira 0.00% Jettenia 0.00% Nitrosomonas 0.00% Brocadia 0.00% Scalindua 0.00% Pirellula 0.00% Gemmata 0.00% Planctomyces 0.43% Candidatus_Nitrosoarchaeum 0.00% Planctomycetaceae 0.00% Isosphera 0.00% Candidatus, Kuenenia 0.00% Anammoxoglobus 0.00% Nitrosococcus 0.00% Nitrosopumilus 0.00% Nitrosocaldus 0.00% Nitrososphaera 0.00% Crenarchaeum 0.00% Nitrite Oxidation Nitrospira 0.00% Nitrobacter 0.06% Nitrospina 0.00% Nitrococcus 0.00% Iron Oxidation Ferribacterium 0.00% Sediminibacterium 0.00% Sideroxydans 0.00% Acidithiobacillus 0.00% Geobacter 8.90% Gallionella 4.21% Leptospirillum 0.00% Crenothrix 7.00% Ferritrophicum 0.00% Ferrovibrio 0.00% Acidiferrobacter 0.00% Sulfate Reduction Desulfuromonas 0.00%

12 Desulfurivibrio 0.00% Desulfovibrio 0.45% Desulfocapsa 0.12% Desulfomonile 3.12% Desulfobacteraceae 0.00% Desulfobulbaceae 0.00% Desulfobacterium 0.00% Desulfosporosinus 0.00% Sulfur Oxidation Thiobacillus 0.00% Thiothrix 0.11% Sulfuritalea 0.21% Sulfuricurvum 0.12% Sulfuricella 0.00% Sulfurospirillum 0.00% Sulfuritalea 0.21% Magnetovibrio 0.00% Methane Oxidation Methanosaeta 0.00% Methanoculleus 0.00% Methyloversatilis 0.08% Methylophilaceae 0.00% Methylosoma 0.00% Methylococcus 0.00% Biofilm Slime Formers Pseudomonas 0.27% Sphingomonas 0.14%

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