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1.
PLoS One ; 15(4): e0232265, 2020.
Article in English | MEDLINE | ID: mdl-32353009

ABSTRACT

The groundwater biome is a poorly characterized habitat hypothesized to harbor uniquely diverse bacterial communities; the degree to which these communities differ from associated soils is a central question in environmental microbiology. We characterized the Bacterial community composition in 37 aquifer and 32 surface soil samples across the island of O'ahu, Hawai'i. Several bacterial phyla (Acetothermia, Omnitrophica, Parcubacteria, Peregrinibacteria) relatively abundant in the aquifer samples were rare to absent in the soils. Immense bacterial diversity detected in the deep aquifers indicates that these environments are not as homogenous as expected, but provide various niches and energy sources for wide variety of bacteria. A small proportion of OTUs were widespread in all the basal (0.63%) and all the dike aquifer (0.31%) samples. However, these core bacteria comprised an average of 31.8% (ranging 16.2%-62.0%) and 15.4% (0.1%-31.5%) of all sequences isolated from the basal and dike aquifers respectively. Bacterial community composition correlated significantly with the sodium, sulfate, potassium, total dissolved solids, nitrate, conductivity, and pH in the basal aquifers, while phosphate and bicarbonate levels were also highly important when dike water samples were included in the analyses. This was consistent with high relative abundance of putative chemolithoautoroph taxa in the aquifer communities relative to soils. Targeted molecular and culture-based fecal indicator microbial analyses indicated good water quality of aquifers. The dominance of unique, deeply branching lineages in tropical aquifers emphasizes a large adaptive potential in O'ahu's aquifers; variability among groundwater samples suggests that aquifer habitats are surprisingly variable potentially harboring a variety of chemolithotrophic energy sources. Although parallel analyses of conventional and alternative indicators indicated good groundwater quality, this study calls for groundwater monitoring programs which would consider public as well as ecosystem health.


Subject(s)
Bacteria/classification , Groundwater/chemistry , Groundwater/microbiology , Ecosystem , Environmental Monitoring/methods , Islands , Nitrates/chemistry , Phylogeny , Sulfates/chemistry , Water Microbiology , Water Quality
2.
Bioresour Technol ; 299: 122554, 2020 Mar.
Article in English | MEDLINE | ID: mdl-31870707

ABSTRACT

The efficacy of biofilm based anaerobic-aerobic treatment to reduce caffeine, carbamazepine, and three estrogens (Estrone (E1), 17ß-estradiol (E2), and 17α-ethynylestradiol (EE2)), as well as E. coli (CN-13) and F+ specific coliphage (MS2), from synthetic wastewater was investigated. Results showed no observable reduction of carbamazepine by either anaerobic or aerobic biofilms over a dosing period of 51-days followed by an additional 23 days of observation. Caffeine, by contrast, was reduced by 11.09% in the upflow anaerobic packed bed biofilm reactor (UAnPBBR) and by 91.90% in the aerobic trickling filter biofilm reactor (TF). Estrone (E1) and 17ß-estradiol (E2) showed minimal reduction in the UAnPBBR but 99.67% reduction in the TF, while EE2 was reduced 1.62% in the AnPBBR and 20.36% in the TF. On average, a 3-log reduction of E. coli (CN-13) and a 1-log reduction of F+ specific coliphage (MS2) concentration was observed across the overall reactor system.


Subject(s)
Wastewater , Water Pollutants, Chemical , Anaerobiosis , Biofilms , Escherichia coli , Estrogens , Estrone , Waste Disposal, Fluid
3.
Water Res ; 116: 23-33, 2017 06 01.
Article in English | MEDLINE | ID: mdl-28292677

ABSTRACT

Indicator bacteria, which are conventionally used to evaluate recreational water quality, can originate from various non-human enteric and extra-enteric sources, hence they may not be indicative of human health risk nor do they provide information on the sources of contamination. In this study we utilized traditional (enterococci and Escherichia coli) and alternative (Clostridium perfringens) indicator bacteria, F+-specific coliphage, molecular markers for microorganisms associated with human sewage (human-associated Bacteroides and polyomaviruses), and microbial community analysis tools (16S rRNA gene fragment amplicon sequencing), to identify and evaluate human sewage-related impact in the Manoa watershed in Honolulu, Hawaii. Elevated concentrations of enterococci (geometric mean ranging from 1604 to 2575 CFU 100 mL-1) and C. perfringens (45-77 CFU 100 mL-1) indicated impairment of the urbanized section of the stream, while indicator bacteria concentrations decreased downstream in the tidally influenced Ala Wai Canal. The threshold values triggering water quality violation notifications in Hawaii were exceeded in 33.3-75.0% of samples collected at sites in the urbanized section of Manoa Stream, but were not exceeded in any of the samples collected at an upstream site located in a forested area. Correlation between indicator bacteria concentrations and rainfall amounts was weak to moderate but significant (E. coli R = 0.251, P = 0.009; enterococci R = 0.369, P < 0.001; C. perfringens R = 0.343, P < 0.001), while concentrations of human fecal-associated molecular markers were not significantly correlated with rainfall (human-associated Bacteroides, R = 0.131, P = 0.256; human-associated polyomaviruses, R = 0.213, P = 0.464). Presence of human sewage was confirmed by detection of human-associated Bacteroides and human polyomavirus in the urbanized section of Manoa Stream (83.3-100% and 41.7-66.7% positive samples respectively). It was further confirmed by microbial community analyses which suggested that an average 2.4-3.4% of the total bacterial population in this section was associated with sewage. Microbial community profiles were significantly influenced by rainfall (R2 = 0.4390, P < 0.001), pH (R2 = 0.3077, P = 0.006), salinity (R2 = 0.2614, P = 0.038), and conductivity (R2 = 0.2676, P = 0.031). Although microbial diversity fluctuated throughout the watershed, it was lower in the impaired section. Leaking sewer systems and illegal cross-connections are implicated in the impairment of the watershed, hence both the sewer and the storm water lines should be routinely inspected. Collectively, our data suggest that information derived from the analysis of microbial communities complements current marker-based microbial source tracking techniques and environmental monitoring programs.


Subject(s)
Sewage/microbiology , Water Quality , Environmental Monitoring , Escherichia coli/genetics , Feces/microbiology , Humans , RNA, Ribosomal, 16S/genetics , Water Microbiology
4.
Environ Sci Pollut Res Int ; 24(13): 12384-12392, 2017 May.
Article in English | MEDLINE | ID: mdl-28357803

ABSTRACT

Roof-harvested rainwater (RHRW) is an important alternative source of water that many island communities can use for drinking and other domestic purposes when groundwater and/or surface water sources are contaminated, limited, or simply not available. The aim of this pilot-scale study was to investigate current RHRW practices in American Samoa (AS) and to evaluate and compare the quality of water from common potable water sources including RHRW stored in tanks, untreated stream water, untreated municipal well water, and treated municipal tap water samples. Samples were analyzed using culture-based methods, quantitative polymerase chain reaction (qPCR), and 16S amplicon sequencing-based methods. Based on indicator bacteria (total coliform and Escherichia coli) concentrations, the quality of RHRW was slightly lower than well and chlorinated tap water but exceeded that of untreated stream water. Although no Giardia or Leptospira spp. were detected in any of the RHRW samples, 86% of the samples were positive for Cryptosporidium spp. All stream water samples tested positive for Cryptosporidium spp. Opportunistic pathogens (Pseudomonas aeruginosa and Mycobacterium intracellulare) were also detected in the RHRW samples (71 and 21% positive samples, respectively). Several potentially pathogenic genera of bacteria were also detected in RHRW by amplicon sequencing. Each RHRW system was characterized by distinct microbial communities, 77% of operational taxonomic units (OTUs) were detected only in a single tank, and no OTU was shared by all the tanks. Risk of water-borne illness increased in the following order: chlorinated tap water/well water < RHRW < stream water. Frequent detection of opportunistic pathogens indicates that RHRW should be treated before use. Stakeholder education on RHRW system design options as well as on importance of regular cleaning and proper management techniques could improve the quality of the RHRW in AS.


Subject(s)
Drinking Water/microbiology , Rain , American Samoa , Bacteria/classification , Humans , Water Microbiology
5.
Appl Environ Microbiol ; 82(22): 6757-6767, 2016 11 15.
Article in English | MEDLINE | ID: mdl-27613686

ABSTRACT

Identification of sources of fecal contaminants is needed to (i) determine the health risk associated with recreational water use and (ii) implement appropriate management practices to mitigate this risk and protect the environment. This study evaluated human-associated Bacteroides spp. (HF183TaqMan) and human polyomavirus (HPyV) markers for host sensitivity and specificity using human and animal fecal samples collected in Hawaii. The decay rates of those markers and indicator bacteria were identified in marine and freshwater microcosms exposed and not exposed to sunlight, followed by field testing of the usability of the molecular markers. Both markers were strongly associated with sewage, although the cross-reactivity of the HF183TaqMan (also present in 82% of canine [n = 11], 30% of mongoose [n = 10], and 10% of feline [n = 10] samples) needs to be considered. Concentrations of HF183TaqMan in human fecal samples exceeded those in cross-reactive animals at least 1,000-fold. In the absence of sunlight, the decay rates of both markers were comparable to the die-off rates of enterococci in experimental freshwater and marine water microcosms. However, in sunlight, the decay rates of both markers were significantly lower than the decay rate of enterococci. While both markers have their individual limitations in terms of sensitivity and specificity, these limitations can be mitigated by using both markers simultaneously; ergo, this study supports the concurrent use of HF183TaqMan and HPyV markers for the detection of sewage contamination in coastal and inland waters in Hawaii. IMPORTANCE: This study represents an in-depth characterization of microbial source tracking (MST) markers in Hawaii. The distribution and concentrations of HF183TaqMan and HPyV markers in human and animal fecal samples and in wastewater, coupled with decay data obtained from sunlight-exposed and unexposed microcosms, support the concurrent application of HF183TaqMan and HPyV markers for sewage contamination detection in Hawaii waters. Both markers are more conservative and more specific markers of sewage than fecal indicator bacteria (enterococci and Escherichia coli). Analysis of HF183TaqMan (or newer derivatives) is recommended for inclusion in future epidemiological studies concerned with beach water quality, while better concentration techniques are needed for HPyV. Such epidemiological studies can be used to develop new recreational water quality criteria, which will provide direct information on the absence or presence of sewage contamination in water samples as well as reliable measurements of the risk of waterborne disease transmission to swimmers.


Subject(s)
Bacteroides/isolation & purification , Feces/microbiology , Polyomavirus/isolation & purification , Sewage/microbiology , Water Microbiology , Adult , Animals , Bacteroides/genetics , Cats , Dogs , Escherichia coli/genetics , Female , Fresh Water/microbiology , Hawaii , Humans , Male , Middle Aged , Polymerase Chain Reaction , Polyomavirus/genetics , Seawater/microbiology , Sunlight , Water Pollution/analysis , Water Quality
6.
Virol Sin ; 30(5): 344-53, 2015 Oct.
Article in English | MEDLINE | ID: mdl-26494480

ABSTRACT

Recreational waters contaminated with human fecal pollution are a public health concern, and ensuring the safety of recreational waters for public use is a priority of both the Environmental Protection Agency (EPA) and the Centers for Disease Control and Prevention (CDC). Current recreational water standards rely on fecal indicator bacteria (FIB) levels as indicators of human disease risk. However present evidence indicates that levels of FIB do not always correspond to the presence of other potentially harmful organisms, such as viruses. Thus, enteric viruses are currently tested as water quality indicators, but have yet to be successfully implemented in routine monitoring of water quality. This study utilized enteric viruses as possible alternative indicators of water quality to examine 18 different fresh and offshore recreational waters on O'ahu, Hawai'i, by using newly established laboratory techniques including highly optimized PCR, real time PCR, and viral infectivity assays. All sample sites were detected positive for human enteric viruses by PCR including enterovirus, norovirus genogroups I and II, and male specific FRNA coliphage. A six time-point seasonal study of enteric virus presence indicated significant variation in virus detection between the rainy and dry seasons. Quantitative PCR detected the presence of norovirus genogroup II at levels at which disease risk may occur, and there was no correlation found between enteric virus presence and FIB counts. Under the present laboratory conditions, no infectious viruses were detected from the samples PCR-positive for enteric viruses. These data emphasize both the need for additional indicators for improved monitoring of water quality, and the feasibility of using enteric viruses as these indicators.


Subject(s)
Enterovirus/isolation & purification , Environmental Monitoring/methods , Fresh Water/virology , Water Microbiology , Water Pollution/analysis , Water Quality/standards , Animals , Centers for Disease Control and Prevention, U.S. , Chlorocebus aethiops , Coliphages/genetics , Enterovirus/genetics , Feces/virology , Hawaii , Humans , Male , Norovirus/genetics , Polymerase Chain Reaction/methods , Real-Time Polymerase Chain Reaction/methods , Recreation , Seasons , United States , United States Environmental Protection Agency , Vero Cells , Water Supply/standards
7.
Int J Environ Res Public Health ; 12(7): 7752-76, 2015 Jul 09.
Article in English | MEDLINE | ID: mdl-26184253

ABSTRACT

This manuscript evaluates the U.S. Recreational Water Quality Criteria (RWQC) of 2012, based upon discussions during a conference held 11-13 March 2013, in Honolulu, Hawaii. The RWQC of 2012 did not meet expectations among the research community because key recommended studies were not completed, new data to assess risks to bathers exposed to non-point sources of fecal indicator bacteria (FIB) were not developed, and the 2012 RWQC did not show marked improvements in strategies for assessing health risks for bathers using all types of recreational waters. The development of the 2012 RWQC was limited in scope because the epidemiologic studies at beach sites were restricted to beaches with point sources of pollution and water samples were monitored for only enterococci. The vision for the future is development of effective RWQC guidelines based on epidemiologic and quantitative microbial risk assessment (QMRA) studies for sewage specific markers, as well as human enteric pathogens so that health risks for bathers at all recreational waters can be determined. The 2012 RWQC introduced a program for states and tribes to develop site-specific water quality criteria, and in theory this approach can be used to address the limitations associated with the measurements of the traditional FIB.


Subject(s)
Recreation , Water Quality , Bacteria/isolation & purification , Bathing Beaches , Enterococcus , Feces/microbiology , Humans , Risk Assessment , Sewage/microbiology , United States , Water Quality/standards
8.
Sci Total Environ ; 524-525: 124-35, 2015 Aug 15.
Article in English | MEDLINE | ID: mdl-25889551

ABSTRACT

Slow sand filtration (SSF) has been widely used as a means of providing potable water due to its efficacy, low cost, and minimal maintenance. Advances in analytical instrumentation have revealed the occurrence of pharmaceutically active compounds (PhACs) in surface water as well as in groundwater. It is unclear if the presence of these compounds in the feed water can interfere with the performances of an SSF unit. The aim of this work was to examine i) the ability of two SSF units to remove six PhACs (caffeine, carbamazepine, 17-ß estradiol [E2], estrone [E1], gemfibrozil, and phenazone), and ii) the impact of these PhACs on the removal of bacteria by two SSF units. The presence of PhACs in feed water for SSF can occur in surface waters impacted by wastewater or leakage from sewers and septic tanks, as well as in developing countries where unregulated use and improper disposal are prevalent. Two pilot-scale SSF units were used during the study. Unit B1 was fed with stream water with 1% of primary effluent added, while unit B2 was fed with stream water alone. Although limited removal (<10%) of carbamazepine, gemfibrozil, and phenazone occurred, the complete removal of caffeine, and the partial removal (11-92%) of E2 and E1 were observed in the two SSF units. The results of this study suggest that the occurrence of the selected PhACs, probably estrogens and caffeine, in the feed water at 50 µg L(-1) affected the ability of the schmutzdecke to remove total coliform and Escherichia coli. The bacterial removal achieved within the schmutzdecke dropped from 95% to less than 20% by the end of the study. This decrease in removal may be related to the change in the microbial community within the schmutzdecke. A diverse microbial community, including Bacteroidetes and several classes of Proteobacteria, was replaced by a microbial community in which Gammaproteobacteria was the predominant phylum (99%). Despite the low removal achieved within the schmutzdecke, removal of total coliform and E. coli greater than 99% occurred after both SSF units throughout the study. Bacterial removal occurred in the upper half of the sand filter. This was probably due to a diverse microbial community established in the packing material, in which Bacteroidetes (13-25%), Acidobacteria (7-17%) and several classes of Proteobacteria (35-52%) (Alpha-, Beta-, Delta-, and Gammaproteobacteria) were the predominant phyla.


Subject(s)
Filtration/methods , Pharmaceutical Preparations/analysis , Waste Disposal, Fluid/methods , Water Pollutants, Chemical/analysis , Biodegradation, Environmental , Silicon Dioxide , Wastewater/chemistry , Water Purification/methods
9.
FEMS Microbiol Ecol ; 76(2): 311-26, 2011 May.
Article in English | MEDLINE | ID: mdl-21255054

ABSTRACT

Development of inhibitors and vaccines that mitigate rumen-derived methane by targeting methanogens relies on knowledge of the methanogens present. We investigated the composition of archaeal communities in the rumens of farmed sheep (Ovis aries), cattle (Bos taurus) and red deer (Cervus elaphus) using denaturing gradient gel electrophoresis (DGGE) to generate fingerprints of archaeal 16S rRNA genes. The total archaeal communities were relatively constant across species and diets, and were less variable and less diverse than bacterial communities. There were diet- and ruminant-species-based differences in archaeal community structure, but the same dominant archaea were present in all rumens. These were members of three coherent clades: species related to Methanobrevibacter ruminantium and Methanobrevibacter olleyae; species related to Methanobrevibacter gottschalkii, Methanobrevibacter thaueri and Methanobrevibacter millerae; and species of the genus Methanosphaera. Members of an archaeal group of unknown physiology, designated rumen cluster C (RCC), were also present. RCC-specific DGGE, clone library analysis and quantitative real-time PCR showed that their 16S rRNA gene sequences were very diverse and made up an average of 26.5% of the total archaea. RCC sequences were not readily detected in the DGGE patterns of total archaeal 16S rRNA genes because no single sequence type was abundant enough to form dominant bands.


Subject(s)
Diet , Methanobacteriaceae/genetics , Rumen/microbiology , Animals , Bacteria/classification , Bacteria/genetics , Cattle/microbiology , DNA, Archaeal/genetics , Deer/microbiology , Denaturing Gradient Gel Electrophoresis , Gene Library , Genes, Archaeal , Genes, Bacterial , Methane , Methanobacteriaceae/classification , Phylogeny , Polymerase Chain Reaction/veterinary , RNA, Bacterial/genetics , RNA, Ribosomal, 16S/genetics , Sheep, Domestic/microbiology
10.
Water Res ; 43(19): 4828-37, 2009 Nov.
Article in English | MEDLINE | ID: mdl-19631958

ABSTRACT

Concentrations of fecal indicator bacteria (FIB; e.g. Escherichia coli, and Enterococcus sp.) can only be used in limited ways for determining the source of fecal contamination in recreational waters because they cannot distinguish human from non-human fecal contamination. Several Bacteroides spp. have been suggested as potential alternative indicators. We have developed a rapid, culture-independent method for quantifying fecal Bacteroides spp. using quantitative PCR (QPCR) targeting the 16S rRNA gene. The assay specifically targets and quantifies the most common human Bacteroides spp. The details of the method are presented, including analyses of a wide range of fecal samples from different organisms. Specificity and performance of the QPCR assay were also tested via a laboratory experiment where human sewage and gull guano were inoculated into a range of environmental water samples. Concentrations of fecal Bacteroides spp., total Enterococcus sp., Enterococcus faecium, Enterococcus faecalis, and Enterococcus casseliflavus were measured using QPCR, and total Enterococcus sp. and E. coli were quantified by membrane filtration (MF). Samples spiked with gull guano were highly concentrated with total Enterococcus sp., E. coli, E. faecalis, and E. casseliflavus, demonstrating that these indicators are prominent in animal feces. On the other hand, fecal Bacteroides spp. concentrations were high in samples containing sewage and were relatively low in samples spiked with gull guano. Sensitivity and specificity results suggest that the rapid fecal Bacteroides spp. QPCR assay may be a useful tool to effectively predict the presence and concentration of human-specific fecal pollution.


Subject(s)
Bacteroidetes/isolation & purification , Feces/microbiology , Polymerase Chain Reaction/methods , Sewage/microbiology , Water Pollutants/isolation & purification , Bacteroidetes/genetics , Enterococcus/genetics , Enterococcus/isolation & purification , Environmental Monitoring , Humans , RNA, Ribosomal, 16S/genetics
11.
Appl Environ Microbiol ; 74(10): 3319-20, 2008 May.
Article in English | MEDLINE | ID: mdl-18378648

ABSTRACT

An assay based on transcription-mediated amplification (TMA) technology was used to quantitate Enterococcus fecal indicator bacteria in environmental water samples. The results generated by this and two growth-based methods relative to the 104 most-probable-number or CFU-per-100-ml threshold show that the three methods are in good qualitative agreement when tested against a range of water samples taken from different locations. The results demonstrate sensitive and rapid detection (approximately 4 h from sample collection to result) and quantitation of Enterococcus bacteria compared to the results with the growth-based methods.


Subject(s)
Bacteriological Techniques/methods , Enterococcus/growth & development , Enterococcus/isolation & purification , Nucleic Acid Amplification Techniques/methods , Water Microbiology , Enterococcus/genetics , Sensitivity and Specificity
13.
Appl Environ Microbiol ; 73(3): 808-14, 2007 Feb.
Article in English | MEDLINE | ID: mdl-17142373

ABSTRACT

It is well documented that microbial contamination of coastal waters poses a significant risk to human health through recreational exposure and consumption of shellfish. Identifying the source of microbial contamination (microbial source tracking) plays a dominant role in enabling effective management and remediation strategies. One method used to determine the source of the contamination is quantification of the ratio of the four subgroups of F+-specific RNA coliphages (family Leviviridae) in impacted water samples. Because of typically low concentrations in the environment, enrichment assays are performed prior to detection, even though differential replication rates have been reported. These assays are also compromised by differential loss of phage infectivity among subgroups after release into the environment, thus obscuring the initial ratio. Here, a culture-independent multiplex real-time reverse transcriptase-PCR (RT-PCR) protocol for the simultaneous quantification of all four subgroups of F+-specific RNA coliphages using novel primer sets and molecular beacons is presented. This assay is extremely sensitive, achieving detection with as few as 10 copies of isolated coliphage RNA, and is linear for a minimum of six orders of magnitude. During survival experiments, the real-time RT-PCR technique was able to quantify coliphages in seawater when culture-based double agar layer assay failed. While infectivity was lost at different rates at the subgroup level, decay constants in seawater, calculated using the real-time RT-PCR estimates, did not vary among subgroups. The accurate determination of the in situ concentration of F+-specific RNA coliphages using this method will facilitate more effective remediation strategies for impacted environments.


Subject(s)
Coliphages/isolation & purification , F Factor/isolation & purification , RNA Phages/isolation & purification , Reverse Transcriptase Polymerase Chain Reaction/methods , Sewage/virology , Water Microbiology , Water Pollution/analysis , Animals , Cats , Cattle , Chickens/virology , Coliphages/genetics , DNA Primers , F Factor/genetics , Feces/virology , Humans , Molecular Probe Techniques , Molecular Sequence Data , RNA Phages/genetics , RNA, Viral/analysis , RNA, Viral/genetics , Sequence Analysis, DNA
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