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Sample records for fecal bacteroidales bacteria

  1. Host Distributions of Uncultivated Fecal Bacteroidales Bacteria Reveal Genetic Markers for Fecal Source Identification

    PubMed Central

    Dick, Linda K.; Bernhard, Anne E.; Brodeur, Timothy J.; Santo Domingo, Jorge W.; Simpson, Joyce M.; Walters, Sarah P.; Field, Katharine G.

    2005-01-01

    The purpose of this study was to examine host distribution patterns among fecal bacteria in the order Bacteroidales, with the goal of using endemic sequences as markers for fecal source identification in aquatic environments. We analyzed Bacteroidales 16S rRNA gene sequences from the feces of eight hosts: human, bovine, pig, horse, dog, cat, gull, and elk. Recovered sequences did not match database sequences, indicating high levels of uncultivated diversity. The analysis revealed both endemic and cosmopolitan distributions among the eight hosts. Ruminant, pig, and horse sequences tended to form host- or host group-specific clusters in a phylogenetic tree, while human, dog, cat, and gull sequences clustered together almost exclusively. Many of the human, dog, cat, and gull sequences fell within a large branch containing cultivated species from the genus Bacteroides. Most of the cultivated Bacteroides species had very close matches with multiple hosts and thus may not be useful targets for fecal source identification. A large branch containing cultivated members of the genus Prevotella included cloned sequences that were not closely related to cultivated Prevotella species. Most ruminant sequences formed clusters separate from the branches containing Bacteroides and Prevotella species. Host-specific sequences were identified for pigs and horses and were used to design PCR primers to identify pig and horse sources of fecal pollution in water. The primers successfully amplified fecal DNAs from their target hosts and did not amplify fecal DNAs from other species. Fecal bacteria endemic to the host species may result from evolution in different types of digestive systems. PMID:15933020

  2. TRACKING FECAL CONTAMINATION WITH BACTEROIDALES MOLECULAR MARKERS: AN ANALYSIS OF THE DYNAMICS OF FECAL CONTAMINATION IN THE TILLAMOOK BASIN, OREGON

    EPA Science Inventory

    Although amplification of source-specific molecular markers from Bacteroidales fecal bacteria can identify several different kinds of fecal contamination in water, it remains unclear how this technique relates to fecal indicator measurements in natural waters. The objectives of t...

  3. TRACKING FECAL CONTAMINATION WITH BACTEROIDALES MOLECULAR MARKERS: AN ANALYSIS OF THE DYNAMICS OF FECAL CONTAMINATION IN THE TILLAMOOK BASIN, OREGON

    EPA Science Inventory

    Although amplification of source-specific molecular markers from Bacteroidales fecal bacteria can identify several different kinds of fecal contamination in water, it remains unclear how this technique relates to fecal indicator measurements in natural waters. The objectives of t...

  4. Distribution of Human-Specific Bacteroidales and Fecal Indicator Bacteria in an Urban Watershed Impacted by Sewage Pollution, Determined Using RNA- and DNA-Based Quantitative PCR Assays

    PubMed Central

    Kapoor, Vikram; Pitkänen, Tarja; Ryu, Hodon; Elk, Michael

    2014-01-01

    The identification of fecal pollution sources is commonly carried out using DNA-based methods. However, there is evidence that DNA can be associated with dead cells or present as “naked DNA” in the environment. Furthermore, it has been shown that rRNA-targeted reverse transcription-quantitative PCR (RT-qPCR) assays can be more sensitive than rRNA gene-based qPCR assays since metabolically active cells usually contain higher numbers of ribosomes than quiescent cells. To this end, we compared the detection frequency of host-specific markers and fecal bacteria using RNA-based RT-qPCR and DNA-based qPCR methods for water samples collected in sites impacted by combined sewer overflows. As a group, fecal bacteria were more frequently detected in most sites using RNA-based methods. Specifically, 8, 87, and 85% of the samples positive for general enterococci, Enterococcus faecalis, and Enterococcus faecium markers, respectively, were detected using RT-qPCR, but not with the qPCR assay counterpart. On average, two human-specific Bacteroidales markers were not detected when using DNA in 12% of the samples, while they were positive for all samples when using RNA (cDNA) as the template. Moreover, signal intensity was up to three orders of magnitude higher in RT-qPCR assays than in qPCR assays. The human-specific Bacteroidales markers exhibited moderate correlation with conventional fecal indicators using RT-qPCR results, suggesting the persistence of nonhuman sources of fecal pollution or the presence of false-positive signals. In general, the results from this study suggest that RNA-based assays can increase the detection sensitivity of fecal bacteria in urban watersheds impacted with human fecal sources. PMID:25326295

  5. DYNAMICS OF AQUATIC FECAL CONTAMINATION, FECAL SOURCE IDENTIFICATION, AND CORRELATION OF BACTEROIDALES HOST-SPECIFIC MARKERS DETECTION WITH FECAL PATHOGENS

    EPA Science Inventory

    Fecal pollution impairs the health and productivity of coastal waters and causes human disease. PCR of host-specific 16S rDNA sequences from anaerobic Bacteroidales bacteria offers a promising method of tracking fecal contamination and identifying its source(s). Before Bacteroida...

  6. DYNAMICS OF AQUATIC FECAL CONTAMINATION, FECAL SOURCE IDENTIFICATION, AND CORRELATION OF BACTEROIDALES HOST-SPECIFIC MARKERS DETECTION WITH FECAL PATHOGENS

    EPA Science Inventory

    Fecal pollution impairs the health and productivity of coastal waters and causes human disease. PCR of host-specific 16S rDNA sequences from anaerobic Bacteroidales bacteria offers a promising method of tracking fecal contamination and identifying its source(s). Before Bacteroida...

  7. Survival and persistence of fecal host-specific Bacteroidales cells and their DNA assessed by PMA-qPCR

    NASA Astrophysics Data System (ADS)

    Bae, S.; Bombardelli, F.; Wuertz, S.

    2008-12-01

    Understanding and managing microbial pollutions in water is one of the foremost challenges of establishing effective managements and remediation strategies to impaired water bodies polluted by uncharacterized fecal sources. Quantitative microbial source tracking (MST) approaches using fecal Bacteroidales and quantitative PCR (qPCR) assays to measure gene copies of host-specific 16S rRNA genetic markers are promising because they can allow for identifying and quantifying fecal loadings from a particular animal host and understanding the fate and transport of host-specific Bacteroidales over a range of conditions in water bodies. Similar to the case of traditional fecal indicator bacteria, a relatively long persistence of target DNA may hamper applied MST studies, if genetic markers cannot be linked to recent fecal pollution in water. We report a successful approach to removing the qPCR signal derived from free DNA and dead host-specific Bacteroidales cells by selectively binding the DNA and consequently inhibiting PCR amplification using light- activated propidium monoazide (PMA). Optimal PMA-qPCR conditions were determined as 100 µM of PMA concentration and a 10-min light exposure time at different solids concentrations in order to mimic a range of water samples. Under these conditions, PMA-qPCR resulted in the selective exclusion of DNA from heat- treated cells of non-culturable Bacteroidales in human feces and wastewater influent and effluent samples. Also, the persistence of feces-derived host-specific Bacteroidales DNA and their cells (determined by universal, human-, cow- and dog-specific Bacteroidales qPCR assays) in seawater was investigated in microcosms at environmental conditions. The average T99 (two log reduction) value for host-specific viable Bacteroidales cells was 28 h, whereas that for total host-specific Bacteroidales DNA was 177 h. Natural sunlight did not have a strong influence on the fate of fecal Bacteroidales cells and their DNA, presumably

  8. [Uncultivated host-specific Bacteroidales markers identification of fecal source pollution--a review].

    PubMed

    Zhang, Xi; Zhu, Changxiong; Zhu, Honghui

    2011-07-01

    Bacteroidales has been proposed as a fecal pollution indicator. microbial source tracking (MST) based on Bacteroidales host-specific gene markers has recently been applied in the fecal pollution identification, which does not require culturing the fecal pollution indicator organisms. This method needs to design specific primers. The primers are designed based on Bacteroidales specific 16S rRNA gene. Once a pair of specific primers was amplified, the fecal pollution can be identified. In this paper, the progress of specific primers of Bacteroidales in human, swine, ruminant feces were reviewed and discussed. The advantages and disadvantages were put forward. Future researchers should be focused on the new biological markers and the combination of different MST methods.

  9. High diversity and differential persistence of fecal Bacteroidales population spiked into freshwater microcosm.

    PubMed

    Liang, Zhanbei; He, Zhenli; Zhou, Xuxia; Powell, Charles A; Yang, Yuangen; Roberts, Michael G; Stoffella, Peter J

    2012-01-01

    Bacteroidales markers are promising indicators of fecal pollution and are now widely used in microbial source tracking (MST) studies. However, a thorough understanding of the persistence of Bacteroidales population after being released into environmental waters is lacking. We investigated the persistence of two host specific markers (HF183 and CF193) and temporal change of Bacteroidales population over 14 days in freshwater microcosms seeded with human or bovine feces. The concentrations of HF183/CF193 and Escherichia coli were determined using quantitative polymerase chain reaction (qPCR) and standard cultivation method, respectively. Shifts in the Bacteroidales population structure were fingerprinted using PCR-denaturing gradient gel electrophoresis (DGGE) and subsequent sequencing analysis targeting both 16S rDNA and rRNA-transcribed cDNA. Both HF183 and CF193 decayed significantly faster than E. coli but the decay curves fit poorly with first-order model. High diversity of Bacteroidales population was observed for both microcosms, and persistence of different species in the population varied. Sequence analysis indicated that most of the bovine Bacteroidales populations in our study are unexplored. DGGE and decay curve indicated that RNA decayed faster than DNA, further supporting the use of rRNA as indicator of metabolically active Bacteroidales population. Evaluations with more realistic scenarios are warranted prior to extending the results of this study to real field settings. Copyright © 2011 Elsevier Ltd. All rights reserved.

  10. Unsuitability of quantitative Bacteroidales 16S rRNA gene assays for discerning fecal contamination of drinking water.

    PubMed

    van der Wielen, Paul W J J; Medema, Gertjan

    2010-07-01

    Bacteroidales species were detected in (tap) water samples from treatment plants with three different PCR assays. 16S rRNA gene sequence analysis indicated that the sequences had an environmental rather than fecal origin. We conclude that assays for Bacteroidales 16S rRNA genes are not specific enough to discern fecal contamination of drinking water in the Netherlands.

  11. Tracing fecal pollution sources in karst groundwater by Bacteroidales genetic biomarkers, bacterial indicators, and environmental variables.

    PubMed

    Zhang, Ya; Kelly, Walton R; Panno, Samuel V; Liu, Wen-Tso

    2014-08-15

    Fecal contamination in Midwestern karst regions was evaluated by simultaneously measuring traditional bacterial indicators (coliforms and Escherichia coli), Bacteroidales-based biomarkers, and environmental variables. Water samples from springs and wells were collected from karst regions in Illinois (IL), Wisconsin (WI), Kentucky (KY), and Missouri (MO). Quantitative PCR (Q-PCR) with seven primer sets targeting different members of Bacteroidales was used to determine the origin of fecal contamination (i.e., from human waste, livestock waste, or both). Most samples were contaminated by both human and animal waste, with a few samples showing pollution solely by one or the other. Spring water tended to have higher levels of contamination than well water, and higher concentrations of fecal biomarkers were detected in urban springs compared to rural spring systems. However, there were discrepancies on contamination profile determined by Bacteroidales-based biomarkers and by traditional bacterial indicators. Among all the environmental parameters examined, E. coli, sulfate, total dissolved solids (TDS), and silicon were significantly correlated (p<0.05) with the level of Bacteroidales-based fecal indicators. A rapid screening method using total nitrogen (TN) and chloride (Cl(-)) concentrations to determine fecal contamination was shown to be effective and correlated well with Bacteroidales-based MST. The results suggest that human and livestock feces co-contaminated a large portion of karst groundwater systems in Midwestern regions, and the inclusion of traditional bacterial indicators, environmental variables, and Bacteroidales-based MST is an effective approach for identifying fecal contamination in karst regions. Copyright © 2014 Elsevier B.V. All rights reserved.

  12. Detection and persistence of fecal Bacteroidales as water quality indicators in unchlorinated drinking water.

    PubMed

    Saunders, Aaron M; Kristiansen, Anja; Lund, Marie B; Revsbech, Niels Peter; Schramm, Andreas

    2009-08-01

    The results of this study support the use of fecal Bacteroidales qPCR as a rapid method to complement traditional, culture-dependent, water quality indicators in systems where drinking water is supplied without chlorination or other forms of disinfection. A SYBR-green based, quantitative PCR assay was developed to determine the concentration of fecal Bacteroidales 16S rRNA gene copies. The persistence of a Bacteroides vulgatus pure culture and fecal Bacteroidales from a wastewater inoculum was determined in unchlorinated drinking water at 10 degrees C. B. vulgatus 16S rRNA gene copies persisted throughout the experimental period (200 days) in sterile drinking water but decayed faster in natural drinking water, indicating that the natural microbiota accelerated decay. In a simulated fecal contamination of unchlorinated drinking water, the decay of fecal Bacteroidales 16S rRNA gene copies was considerably faster than the pure culture but similar to that of Escherichia coli from the same wastewater inoculum.

  13. Molecular Diversity of Bacteroidales in Fecal and Environmental Samples and Swine-Associated Subpopulations

    EPA Science Inventory

    Several swine-specific microbial source tracking methods are based on PCR assays targeting Bacteroidales 16S rRNA gene sequences. The limited application of these assays can be explained by the poor understanding of their molecular diversity in fecal sources and environmental wat...

  14. Molecular Diversity of Bacteroidales in Fecal and Environmental Samples and Swine-Associated Subpopulations

    PubMed Central

    Lamendella, Regina; Li, Kent C.; Oerther, Daniel

    2013-01-01

    Several swine-specific microbial source tracking methods are based on PCR assays targeting Bacteroidales 16S rRNA gene sequences. The limited application of these assays can be explained by the poor understanding of their molecular diversity in fecal sources and environmental waters. In order to address this, we studied the diversity of 9,340 partial (>600 bp in length) Bacteroidales 16S rRNA gene sequences from 13 fecal sources and nine feces-contaminated watersheds. The compositions of major Bacteroidales populations were analyzed to determine which host and environmental sequences were contributing to each group. This information allowed us to identify populations which were both exclusive to swine fecal sources and detected in swine-contaminated waters. Phylogenetic and diversity analyses revealed that some markers previously believed to be highly specific to swine populations are shared by multiple hosts, potentially explaining the cross-amplification signals obtained with nontargeted hosts. These data suggest that while many Bacteroidales populations are cosmopolitan, others exhibit a preferential host distribution and may be able to survive different environmental conditions. This study further demonstrates the importance of elucidating the diversity patterns of targeted bacterial groups to develop more inclusive fecal source tracking applications. PMID:23160126

  15. Molecular Diversity of Bacteroidales in Fecal and Environmental Samples and Swine-Associated Subpopulations

    EPA Science Inventory

    Several swine-specific microbial source tracking methods are based on PCR assays targeting Bacteroidales 16S rRNA gene sequences. The limited application of these assays can be explained by the poor understanding of their molecular diversity in fecal sources and environmental wat...

  16. Identifying human and livestock sources of fecal contamination in Kenya with host-specific Bacteroidales assays.

    PubMed

    Jenkins, Marion W; Tiwari, Sangam; Lorente, Mario; Gichaba, Charles Maina; Wuertz, Stefan

    2009-11-01

    Microbial source tracking to distinguish between human, livestock and wildlife fecal pollution using molecular techniques is a rapidly evolving approach in many developed countries, but has not previously been applied on the African continent. DNA extracts from cow, donkey, and human fecal specimens and raw domestic sewage samples collected in Kenya were tested against five existing quantitative PCR assays designed to detect universal (2), human-specific (2), and cow-specific (1) fecal Bacteroidales genetic markers. Water samples from the River Njoro in Kenya were evaluated using the five tested Bacteroidales markers and a multi-species assay for Cryptosporidium in a preliminary exploration of fecal pollution sources and health risks in this watershed. Diagnostic sensitivity on the validation set varied from 18 to 100% for the five assays while diagnostic specificity was 100%. Of the 2 universal assays, Total Bacteroidales [Dick, L.K, Field, K.G., 2004. Rapid estimation of numbers of fecal Bacteroidetes by use of a quantitative PCR assay for 16S rRNA genes. Appl. Environ. Microbiol. 70, 5695-5697] showed lower generic fecal diagnostic sensitivity, at 55%, than BacUni-UCD, at 100%, in detecting fecal markers on the 42-sample validation set. Human-specific assay HF183 demonstrated 65% sensitivity overall, and 80% on the human sewage samples, compared to 18% overall and 0% sewage for human-specific assay BacHum-UCD. Cow-specific assay BacCow-UCD had 94% sensitivity. Testing of 18 water samples indicates cows are a likely predominant source of fecal contamination in the Njoro Watershed (78% prevailing rate). Probabilistic assessment of human assay results indicates at most three of the river water samples contained human Bacteroidales. Cryptosporidium spp. markers were detected in samples from nine of the 12 sampling locations. Evidence suggesting widespread contamination by cow feces and Cryptosporidium in the Njoro watershed raises serious concerns for human and

  17. Lachnospiraceae and Bacteroidales Alternative Fecal Indicators Reveal Chronic Human Sewage Contamination in an Urban Harbor▿†

    PubMed Central

    Newton, Ryan J.; VandeWalle, Jessica L.; Borchardt, Mark A.; Gorelick, Marc H.; McLellan, Sandra L.

    2011-01-01

    The complexity of fecal microbial communities and overlap among human and other animal sources have made it difficult to identify source-specific fecal indicator bacteria. However, the advent of next-generation sequencing technologies now provides increased sequencing power to resolve microbial community composition within and among environments. These data can be mined for information on source-specific phylotypes and/or assemblages of phylotypes (i.e., microbial signatures). We report the development of a new genetic marker for human fecal contamination identified through microbial pyrotag sequence analysis of the V6 region of the 16S rRNA gene. Sequence analysis of 37 sewage samples and comparison with database sequences revealed a human-associated phylotype within the Lachnospiraceae family, which was closely related to the genus Blautia. This phylotype, termed Lachno2, was on average the second most abundant fecal bacterial phylotype in sewage influent samples from Milwaukee, WI. We developed a quantitative PCR (qPCR) assay for Lachno2 and used it along with the qPCR-based assays for human Bacteroidales (based on the HF183 genetic marker), total Bacteroidales spp., and enterococci and the conventional Escherichia coli and enterococci plate count assays to examine the prevalence of fecal and human fecal pollution in Milwaukee's harbor. Both the conventional fecal indicators and the human-associated indicators revealed chronic fecal pollution in the harbor, with significant increases following heavy rain events and combined sewer overflows. The two human-associated genetic marker abundances were tightly correlated in the harbor, a strong indication they target the same source (i.e., human sewage). Human adenoviruses were routinely detected under all conditions in the harbor, and the probability of their occurrence increased by 154% for every 10-fold increase in the human indicator concentration. Both Lachno2 and human Bacteroidales increased specificity to

  18. Use of Bacteroidales microbial source tracking to monitor fecal contamination in fresh produce production.

    PubMed

    Ravaliya, Kruti; Gentry-Shields, Jennifer; Garcia, Santos; Heredia, Norma; Fabiszewski de Aceituno, Anna; Bartz, Faith E; Leon, Juan S; Jaykus, Lee-Ann

    2014-01-01

    In recent decades, fresh and minimally processed produce items have been associated with an increasing proportion of food-borne illnesses. Most pathogens associated with fresh produce are enteric (fecal) in origin, and contamination can occur anywhere along the farm-to-fork chain. Microbial source tracking (MST) is a tool developed in the environmental microbiology field to identify and quantify the dominant source(s) of fecal contamination. This study investigated the utility of an MST method based on Bacteroidales 16S rRNA gene sequences as a means of identifying potential fecal contamination, and its source, in the fresh produce production environment. The method was applied to rinses of fresh produce, source and irrigation waters, and harvester hand rinses collected over the course of 1 year from nine farms (growing tomatoes, jalapeño peppers, and cantaloupe) in Northern Mexico. Of 174 samples, 39% were positive for a universal Bacteroidales marker (AllBac), including 66% of samples from cantaloupe farms (3.6 log10 genome equivalence copies [GEC]/100 ml), 31% of samples from tomato farms (1.7 log10 GEC/100 ml), and 18% of samples from jalapeño farms (1.5 log10 GEC/100 ml). Of 68 AllBac-positive samples, 46% were positive for one of three human-specific markers, and none were positive for a bovine-specific marker. There was no statistically significant correlation between Bacteroidales and generic Escherichia coli across all samples. This study provides evidence that Bacteroidales markers may serve as alternative indicators for fecal contamination in fresh produce production, allowing for determination of both general contamination and that derived from the human host.

  19. Microplate subtractive hybridization to enrich for bacteroidales genetic markers for fecal source identification.

    PubMed

    Dick, Linda K; Simonich, Michael T; Field, Katharine G

    2005-06-01

    The ability to identify sources of fecal pollution plays a key role in the analysis of human health risk and the implementation of water resource management strategies. One approach to this problem involves the identification of bacterial lineages or gene sequences that are found exclusively in a particular host species or group. We used subtractive hybridization to enrich for target host-specific fecal Bacteroidales rRNA gene fragments that were different from those of very closely related reference (subtracter) host sources. Target host rRNA gene fragments were hybridized to subtracter rRNA gene fragments immobilized in a microplate well, and target sequences that did not hybridize were cloned and sequenced for PCR primer design. The use of microplates for DNA immobilization resulted in a one-step subtractive hybridization in which the products could be directly amplified with PCR. The new host-specific primers designed from subtracted target fragments differentiated among very closely related Bacteroidales rRNA gene sequences and distinguished between similar fecal sources, such as elk and cow or human and domestic pet (dog).

  20. Phylogenetic Analysis of Bacteroidales 16S rRNA Genes Unveils Sequences Specific to Diverse Swine Fecal Sources

    EPA Science Inventory

    Two of the currently available methods to assess swine fecal pollution (Bac1 and PF163) target Bacteroidales 16S rRNA genes. However, these assays have been shown to exhibit poor host-specificity and low detection limits in environmental waters, in part due to the limited number...

  1. Phylogenetic Analysis of Bacteroidales 16S rRNA Genes Unveils Sequences Specific to Diverse Swine Fecal Sources

    EPA Science Inventory

    Two of the currently available methods to assess swine fecal pollution (Bac1 and PF163) target Bacteroidales 16S rRNA genes. However, these assays have been shown to exhibit poor host-specificity and low detection limits in environmental waters, in part due to the limited number...

  2. The fecal bacteria

    USGS Publications Warehouse

    Sadowsky, Michael J.; Whitman, Richard L.

    2011-01-01

    The Fecal Bacteria offers a balanced, integrated discussion of fecal bacteria and their presence and ecology in the intestinal tract of mammals, in the environment, and in the food supply. This volume covers their use in examining and assessing water quality in order to offer protection from illnesses related to swimming in or ingesting contaminated water, in addition to discussing their use in engineering considerations of water quality, modeling, monitoring, and regulations. Fecal bacteria are additionally used as indicators of contamination of ready-to-eat foods and fresh produce. The intestinal environment, the microbial community structure of the gut microbiota, and the physiology and genomics of this broad group of microorganisms are explored in the book. With contributions from an internationally recognized group of experts, the book integrates medicine, public health, environmental, and microbiological topics in order to provide a unique, holistic understanding of fecal bacteria. Moreover, it shows how the latest basic science and applied research findings are helping to solve problems and develop effective management strategies. For example, readers will discover how the latest tools and molecular approaches have led to our current understanding of fecal bacteria and enabled us to improve human health and water quality. The Fecal Bacteria is recommended for microbiologists, clinicians, animal scientists, engineers, environmental scientists, food safety experts, water quality managers, and students. It will help them better understand fecal bacteria and use their knowledge to protect human and environmental health. They can also apply many of the techniques and molecular tools discussed in this book to the study of a broad range of microorganisms in a variety of habitats.

  3. Lachnospiraceae- and Bacteroidales alternative fecal indicators reveal chronic human sewage contamination in an urban harbor

    USDA-ARS?s Scientific Manuscript database

    The complexity of fecal microbial communities and overlap among human and other animal sources has made it difficult to identify source-specific fecal indicator bacteria. However, the advent of next-generation sequencing technologies has provided increased power to resolve microbial community compos...

  4. Application of leftover sample material from waterborne protozoa monitoring for the molecular detection of Bacteroidales and fecal source tracking markers

    EPA Science Inventory

    In this study, we examined the potential for detecting fecal bacteria and microbial source tracking markers in samples discarded during the concentration of Cryptosporidium and Giardia using USEPA Method 1623. Recovery rates for different fecal bacteria were determined using sp...

  5. Application of leftover sample material from waterborne protozoa monitoring for the molecular detection of Bacteroidales and fecal source tracking markers

    EPA Science Inventory

    In this study, we examined the potential for detecting fecal bacteria and microbial source tracking markers in samples discarded during the concentration of Cryptosporidium and Giardia using USEPA Method 1623. Recovery rates for different fecal bacteria were determined using sp...

  6. Evidence for Extraintestinal Growth of Bacteroidales Originating from Poultry Litter

    PubMed Central

    Mantha, Sirisha; Hair, Elliott; Nayak, Bina; Harwood, Valerie J.

    2014-01-01

    Water quality monitoring techniques that target microorganisms in the order Bacteroidales are potential alternatives to conventional methods for detection of fecal indicator bacteria. Bacteroidales and members of the genus Bacteroides have been the focus of microbial source tracking (MST) investigations for discriminating sources of fecal pollution (e.g., human or cattle feces) in environmental waters. For accurate source apportionment to occur, one needs to understand both the abundance of Bacteroides in host feces and the survival of these host-associated microbial markers after deposition in the environment. Studies were undertaken to evaluate the abundance, persistence, and potential for growth of Bacteroidales originating from poultry litter under oxic and anoxic environmental conditions. Bacteroidales abundance, as determined by quantitative PCR (qPCR) with GenBac primers and probe, increased 2 to 5 log gene copies ml−1 and 2 log gene copies g litter−1 under most conditions during incubation of poultry litter in a variety of laboratory microcosm and field mesocosm studies. DNA sequencing of the Bacteroidales organisms in the litter identified taxa with sequences corresponding exactly to the GenBac primer and probe sequences and that were closely related to Bacteroides uniformis, B. ovatus, and B. vulgatus. These results suggest that MST studies using qPCR methods targeting Bacteroidales in watersheds that are affected by poultry litter should be interpreted cautiously. Growth of Bacteroidales originating from poultry litter in environmental waters may occur while Bacteroidales growth from other fecal sources declines, thus confounding the interpretation of MST results. PMID:25326306

  7. Evaluation of Swine-Specific PCR Assays Used for Fecal Source Tracking and Analysis of Molecular Diversity of Swine-Specific "Bacteroidales" Populations

    EPA Science Inventory

    In this study we evaluated specificity, distribution, and sensitivity of Bacteroidales – (PF163 and PigBac1) and methanogen-based (P23-2) assays proposed to detect swine fecal pollution in environmental waters. The assays were tested against 220 fecal DNA extracts derived from t...

  8. Evaluation of Swine-Specific PCR Assays Used for Fecal Source Tracking and Analysis of Molecular Diversity of Swine-Specific "Bacteroidales" Populations

    EPA Science Inventory

    In this study we evaluated specificity, distribution, and sensitivity of Bacteroidales – (PF163 and PigBac1) and methanogen-based (P23-2) assays proposed to detect swine fecal pollution in environmental waters. The assays were tested against 220 fecal DNA extracts derived from t...

  9. Validation of Bacteroidales quantitative PCR assays targeting human and animal fecal contamination in the public and domestic domains in India.

    PubMed

    Odagiri, Mitsunori; Schriewer, Alexander; Hanley, Kaitlyn; Wuertz, Stefan; Misra, Pravas R; Panigrahi, Pinaki; Jenkins, Marion W

    2015-01-01

    We compared host-associated Bacteroidales qPCR assays developed in the continental United States and Europe for the purpose of measuring the effect of improved sanitation on human fecal exposure in rural Indian communities where both human and animal fecal loading are high. Ten candidate Bacteroidales qPCR assays were tested against fecal samples (human, sewage, cow, buffalo, goat, sheep, dog and chicken) from a test set of 30 individual human, 5 sewage, and 60 pooled animal samples collected in coastal Odisha, India. The two universal/general Bacteroidales assays tested (BacUni, GenBac3) performed equally well, achieving 100% sensitivity on the test set. Across the five human-associated assays tested (HF183 Taqman, BacHum, HumM2, BacH, HF183 SYBR), we found low sensitivity (17 to 49%) except for HF183 SYBR (89%), and moderate to high cross-reactivity with dog (20 to 80%) and chicken fecal samples (60 to 100%). BacHum had the highest accuracy (67%), amplified all sewage samples within the range of quantification (ROQ), and did not cross-react with any fecal samples from cows, the most populous livestock animal in India. Of the ruminant- and cattle-associated assays tested (BacCow, CowM2), BacCow was more sensitive in detecting the full range of common Indian livestock animal fecal sources, while CowM2 only detected cow sources with 50% sensitivity. Neither assay cross-reacted with human sources. BacCan, the dog-associated assay tested, showed no cross-reactivity with human sources, and high sensitivity (90%) for dog fecal samples. Overall, our results indicate BacUni, BacHum, HumM2, BacCan and BacCow would be the most suitable MST assays to distinguish and quantify relative amounts of human-associated and livestock/domestic animal-associated contributions to fecal contamination in Odisha, India. Copyright © 2014 Elsevier B.V. All rights reserved.

  10. Quantitative Real-Time PCR Analysis of Total Propidium Monazide -Resistant Fecal Indicator Bacteria in Wastewater

    EPA Science Inventory

    A real-time quantitative PCR (qPCR) method and a modification of this method incorporating pretreatment of samples with propidium monoazide (PMA) were evaluated for respective analyses of total and presumptively viable Enterococcus and Bacteroidales fecal indicator bacteria. Thes...

  11. Quantitative Real-Time PCR Analysis of Total Propidium Monazide -Resistant Fecal Indicator Bacteria in Wastewater

    EPA Science Inventory

    A real-time quantitative PCR (qPCR) method and a modification of this method incorporating pretreatment of samples with propidium monoazide (PMA) were evaluated for respective analyses of total and presumptively viable Enterococcus and Bacteroidales fecal indicator bacteria. Thes...

  12. Assessment of a new Bacteroidales marker targeting North American beaver (Castor canadensis) fecal pollution by real-time PCR.

    PubMed

    Marti, Romain; Zhang, Yun; Tien, Yuan-Ching; Lapen, David R; Topp, Edward

    2013-11-01

    In many settings wildlife can be a significant source of fecal pathogen input into surface water. The North American beaver (Castor canadensis) is a zoonotic reservoir for several human pathogens including Cryptosporidium spp. and Giardia spp. In order to specifically detect fecal pollution by beavers, we have developed and validated a beaver-specific Bacteroidales marker, designated Beapol01, based on the 16S rRNA gene. The marker is suitable for quantifying pollution using real-time PCR. The specificity and sensitivity of the marker was excellent, Beaver signal was detected in water of a mixed-activity watershed harbouring this rodent. Overall, Beapol01 will be useful for a better understanding of fecal source inputs in drainage basins inhabited by the beaver. © 2013.

  13. Decay of Bacteroidales genetic markers in relation to traditional fecal indicators for water quality modeling of drinking water sources.

    PubMed

    Sokolova, Ekaterina; Aström, Johan; Pettersson, Thomas J R; Bergstedt, Olof; Hermansson, Malte

    2012-01-17

    The implementation of microbial fecal source tracking (MST) methods in drinking water management is limited by the lack of knowledge on the transport and decay of host-specific genetic markers in water sources. To address these limitations, the decay and transport of human (BacH) and ruminant (BacR) fecal Bacteroidales 16S rRNA genetic markers in a drinking water source (Lake Rådasjön in Sweden) were simulated using a microbiological model coupled to a three-dimensional hydrodynamic model. The microbiological model was calibrated using data from outdoor microcosm trials performed in March, August, and November 2010 to determine the decay of BacH and BacR markers in relation to traditional fecal indicators. The microcosm trials indicated that the persistence of BacH and BacR in the microcosms was not significantly different from the persistence of traditional fecal indicators. The modeling of BacH and BacR transport within the lake illustrated that the highest levels of genetic markers at the raw water intakes were associated with human fecal sources (on-site sewers and emergency sewer overflow). This novel modeling approach improves the interpretation of MST data, especially when fecal pollution from the same host group is released into the water source from different sites in the catchment.

  14. Phylogenetic Diversity and Membership Patterns of Fecal and Environmental Bacteroidales Populations

    EPA Science Inventory

    Members of the Bacteroidales order have recently been targets of microbial source tracking assays. While several studies have documented that some populations exhibit preferential host-distribution, given the vast diversity of this bacterial group, it is still necessary to resolv...

  15. Phylogenetic Diversity and Membership Patterns of Fecal and Environmental Bacteroidales Populations

    EPA Science Inventory

    Members of the Bacteroidales order have recently been targets of microbial source tracking assays. While several studies have documented that some populations exhibit preferential host-distribution, given the vast diversity of this bacterial group, it is still necessary to resolv...

  16. Analysis of Enterococci and Bacteriodales Fecal Indicator Bacteria in a Lake Michigan Tributary by Real-Time Quantitative PCR

    EPA Science Inventory

    The Salt Creek watershed in northwest Indiana drains into Lake Michigan near several heavily used recreational beaches. This study aimed to investigate the levels of fecal indicator bacteria, enterococci and Bacteroidales, in Salt Creek using real-time quantitative PCR (qPCR) an...

  17. Analysis of Enterococci and Bacteriodales Fecal Indicator Bacteria in a Lake Michigan Tributary by Real-Time Quantitative PCR

    EPA Science Inventory

    The Salt Creek watershed in northwest Indiana drains into Lake Michigan near several heavily used recreational beaches. This study aimed to investigate the levels of fecal indicator bacteria, enterococci and Bacteroidales, in Salt Creek using real-time quantitative PCR (qPCR) an...

  18. Evaluation of genetic markers from the 16S rRNA gene V2 region for use in quantitative detection of selected Bacteroidales species and human fecal waste by real time PCR

    EPA Science Inventory

    Molecular methods for rapidly quantifying defined Bacteroidales species from the human gastrointestinal tract may have important clinical and environmental applications, ranging from diagnosis of infections to fecal source tracking in surface waters. In this study, sequences from...

  19. Evaluation of genetic markers from the 16S rRNA gene V2 region for use in quantitative detection of selected Bacteroidales species and human fecal waste by real time PCR

    EPA Science Inventory

    Molecular methods for rapidly quantifying defined Bacteroidales species from the human gastrointestinal tract may have important clinical and environmental applications, ranging from diagnosis of infections to fecal source tracking in surface waters. In this study, sequences from...

  20. Applicability of universal Bacteroidales genetic marker for microbial monitoring of drinking water sources in comparison to conventional indicators.

    PubMed

    Shahryari, A; Nikaeen, M; Khiadani Hajian, M; Nabavi, F; Hatamzadeh, M; Hassanzadeh, A

    2014-11-01

    Water quality monitoring is essential for the provision of safe drinking water. In this study, we compared a selection of fecal indicators with universal Bacteroidales genetic marker to identify fecal pollution of a variety of drinking water sources. A total of 60 samples were collected from water sources. The microbiological parameters included total coliforms, fecal coliforms, Escherichia coli and fecal streptococci as the fecal indicator bacteria (FIB), Clostridium perfringens and H2S bacteria as alternative indicators, universal Bacteroidales genetic marker as a promising alternative fecal indicator, and Salmonella spp., Shigella spp., and E. coli O157 as pathogenic bacteria. From 60 samples analyzed, Bacteroidales was the most frequently detected indicator followed by total coliforms. However, the Bacteroidales assay failed to detect the marker in nine samples positive for FIB and other alternative indicators. The results of our study showed that the absence of Bacteroidales is not necessarily an evidence of fecal and pathogenic bacteria absence and may be unable to ensure the safety of the water. Further research, however, is required for a better understanding of the use of a Bacteroidales genetic marker as an indicator in water quality monitoring programs.

  1. Tracking the relative concentration between Bacteroidales DNA markers and culturable Escherichia coli in fecally polluted subtropical seawater: potential use in differentiating fresh and aged pollution.

    PubMed

    Liu, Rulong; Yeung, Leo T C; Ho, Pui-Hei; Lau, Stanley C K

    2017-03-01

    Routine water quality monitoring practices based on the enumeration of culturable Escherichia coli provides no information about the source or age of fecal pollution. An emerging strategy is to use culturable E. coli and the DNA markers of Bacteroidales complementarily for microbial source tracking. In this study, we consistently observed in seawater microcosms of 3 different conditions that culturable E. coli decayed faster (T99 = 1.14 - 4.29 days) than Bacteroidales DNA markers did (T99 = 1.81 - 200.23 days). Concomitantly, the relative concentration between Bacteroidales DNA markers and culturable E. coli increased over time in all treatments. Particularly, the increase during the early stage of the experiments (before T99 of E. coli was reached) was faster than during the later stage (after T99 of E. coli was attained). We propose that the tracking of the relative concentration between Bacteroidales DNA markers and culturable E. coli provides an opportunity to differentiate a pollution that is relatively fresh from one that has aged. This method, upon further investigation and validation, could be useful in episodic pollution events where the surge of E. coli concentration causes noncompliance to the single sample maximum criterion that mandates high frequency follow-up monitoring.

  2. Performance of PCR-based assays targeting Bacteroidales genetic markers of human fecal pollution in sewage and fecal samples

    EPA Science Inventory

    There are numerous PCR-based methods available to characterize human fecal pollution in ambient waters. Each assay employs distinct oligonucleotides and many target different genes and microorganisms leading to potential variations in method performance. Laboratory comparisons ...

  3. Quantitative identification of fecal water pollution sources by TaqMan real-time PCR assays using Bacteroidales 16S rRNA genetic markers.

    PubMed

    Lee, Dae-Young; Weir, Susan C; Lee, Hung; Trevors, Jack T

    2010-12-01

    PCR-based analysis of Bacteroidales 16S rRNA genes has emerged as a promising tool to identify sources of fecal water pollution. In this study, three TaqMan real-time PCR assays (BacGeneral, BacHuman, and BacBovine) were developed and evaluated for their ability to quantitatively detect general (total), human-specific, and bovine-specific Bacteroidales 16S rRNA genetic markers. The detection sensitivity was determined to be 6.5 copies of 16S rRNA gene for the BacGeneral and BacHuman assays and 10 copies for the BacBovine assay. The assays were capable of detecting approximately one to two cells per PCR. When tested with 70 fecal samples from various sources (human, cattle, pig, deer, dog, cat, goose, gull, horse, and raccoon), the three assays positively identified the target markers in all samples without any false-negative results. The BacHuman and BacBovine assays exhibited false-positive reactions with non-target samples in a few cases. However, the level of the false-positive reactions was about 50 times smaller than that of the true-positive ones, and therefore, these cross-reactions were unlikely to cause misidentifications of the fecal pollution sources. Microbial source-tracking capability was tested at two freshwater streams of which water quality was influenced by human and cattle feces, respectively. The assays accurately detected the presence of the corresponding host-specific markers upon fecal pollution and the persistence of the markers in downstream areas. The assays are expected to reliably determine human and bovine fecal pollution sources in environmental water samples.

  4. Relevance of Bacteroidales and F-Specific RNA Bacteriophages for Efficient Fecal Contamination Tracking at the Level of a Catchment in France

    PubMed Central

    Mauffret, Aourell; Caprais, Marie-Paule

    2012-01-01

    The relevance of three host-associated Bacteroidales markers (HF183, Rum2Bac, and Pig2Bac) and four F-specific RNA bacteriophage genogroups (FRNAPH I to IV) as microbial source tracking markers was assessed at the level of a catchment (Daoulas, France). They were monitored together with fecal indicators (Escherichia coli and enterococci) and chemophysical parameters (rainfall, temperature, salinity, pH, and turbidity) by monthly sampling over 2 years (n = 240 water samples) and one specific sampling following an accidental pig manure spillage (n = 5 samples). During the 2-year regular monitoring, levels of E. coli, enterococci, total F-specific RNA bacteriophages, and the general Bacteroidales marker AllBac were strongly correlated with one another and with Rum2Bac (r = 0.37 to 0.50, P < 0.0001). Their correlations with HF183 and FRNAPH I and II were lower (r = 0.21 to 0.29, P < 0.001 to P < 0.0001), and HF183 and enterococci were associated rather than correlated (Fisher's exact test, P < 0.01). Rum2Bac and HF183 enabled 73% of water samples that had ≥2.7 log10 most probably number (MPN) of E. coli/100 ml to be classified. FRNAPH I and II enabled 33% of samples at this contamination level to be classified. FRNAPH I and II complemented the water sample classification obtained with the two Bacteroidales markers by an additional 8%. Pig2Bac and FRNAPH III and IV were observed in a small number of samples (n = 0 to 4 of 245). The present study validates Rum2Bac and HF183 as relevant tools to trace fecal contamination originating from ruminant or human waste, respectively, at the level of a whole catchment. PMID:22610433

  5. Relevance of Bacteroidales and F-specific RNA bacteriophages for efficient fecal contamination tracking at the level of a catchment in France.

    PubMed

    Mauffret, Aourell; Caprais, Marie-Paule; Gourmelon, Michèle

    2012-08-01

    The relevance of three host-associated Bacteroidales markers (HF183, Rum2Bac, and Pig2Bac) and four F-specific RNA bacteriophage genogroups (FRNAPH I to IV) as microbial source tracking markers was assessed at the level of a catchment (Daoulas, France). They were monitored together with fecal indicators (Escherichia coli and enterococci) and chemophysical parameters (rainfall, temperature, salinity, pH, and turbidity) by monthly sampling over 2 years (n = 240 water samples) and one specific sampling following an accidental pig manure spillage (n = 5 samples). During the 2-year regular monitoring, levels of E. coli, enterococci, total F-specific RNA bacteriophages, and the general Bacteroidales marker AllBac were strongly correlated with one another and with Rum2Bac (r = 0.37 to 0.50, P < 0.0001). Their correlations with HF183 and FRNAPH I and II were lower (r = 0.21 to 0.29, P < 0.001 to P < 0.0001), and HF183 and enterococci were associated rather than correlated (Fisher's exact test, P < 0.01). Rum2Bac and HF183 enabled 73% of water samples that had ≥ 2.7 log(10) most probably number (MPN) of E. coli/100 ml to be classified. FRNAPH I and II enabled 33% of samples at this contamination level to be classified. FRNAPH I and II complemented the water sample classification obtained with the two Bacteroidales markers by an additional 8%. Pig2Bac and FRNAPH III and IV were observed in a small number of samples (n = 0 to 4 of 245). The present study validates Rum2Bac and HF183 as relevant tools to trace fecal contamination originating from ruminant or human waste, respectively, at the level of a whole catchment.

  6. 16S rRNA-based assays for quantitative detection of universal, human-, cow-, and dog-specific fecal Bacteroidales: a Bayesian approach.

    PubMed

    Kildare, Beverly J; Leutenegger, Christian M; McSwain, Belinda S; Bambic, Dustin G; Rajal, Veronica B; Wuertz, Stefan

    2007-08-01

    We report the design and validation of new TaqMan((R)) assays for microbial source tracking based on the amplification of fecal 16S rRNA marker sequences from uncultured cells of the order Bacteroidales. The assays were developed for the detection and enumeration of non-point source input of fecal pollution to watersheds. The quantitative "universal"Bacteroidales assay BacUni-UCD detected all tested stool samples from human volunteers (18 out of 18), cat (7 out of 7), dog (8 out of 8), seagull (10/10), cow (8/8), horse (8/8), and wastewater effluent (14/14). The human assay BacHum-UCD discriminated fully between human and cow stool samples but did not detect all stool samples from human volunteers (12/18). In addition, there was 12.5% detection of dog stool (1/8), but no cross-reactivity with cat, horse, or seagull fecal samples. In contrast, all wastewater samples were positive for the BacHum-UCD marker, supporting its designation as 100% sensitive for mixed-human source identification. The cow-specific assay BacCow-UCD fully discriminated between cow and human stool samples. There was 38% detection of horse stool (3/8), but no cross-specificity with any of the other animal stool samples tested. The dog assay BacCan-UCD discriminated fully between dog and cow stool or seagull guano samples and detected 62.5% stool samples from dogs (5/8). There was some cross-reactivity with 22.2% detection of human stool (4/18), 14.3% detection of cat stool (1/7), and 28.6% detection of wastewater samples (4/14). After validation using stool samples, single-blind tests were used to further demonstrate the efficacy of the developed markers; all assays were sensitive, reproducible, and accurate in the quantification of mixed fecal sources present in aqueous samples. Finally, the new assays were compared with previously published sequences, which showed the new methodologies to be more specific and sensitive. Using Bayes' Theorem, we calculated the conditional probability that the

  7. Environmental sources of fecal bacteria

    USGS Publications Warehouse

    Byappanahalli, Muruleedhara N.; Ishii, Satoshi; Sadowsky, Michael J.; Whitman, Richard L.

    2011-01-01

    This chapter provides a review of the research on environmental occurrences of faecal indicator bacteria in a variety of terrestrial and aquatic habitats under different geographic and climatic conditions, and discusses how these external sources may affect surface water quality.

  8. Quantitative real-time PCR analysis of total and propidium monoazide-resistant fecal indicator bacteria in wastewater.

    PubMed

    Varma, M; Field, R; Stinson, M; Rukovets, B; Wymer, L; Haugland, R

    2009-11-01

    A real-time quantitative PCR (qPCR) method and a modification of this method incorporating pretreatment of samples with propidium monoazide (PMA) were evaluated for respective analyses of total and presumptively viable Enterococcus and Bacteroidales fecal indicator bacteria. These methods were used in the analyses of wastewater samples to investigate their feasibility as alternatives to current fecal indicator bacteria culture methods for predicting the efficiency of viral pathogen removal by standard treatment processes. PMA treatment was effective in preventing qPCR detection of target sequences from non-viable cells. Concentrates of small volume, secondary-treated wastewater samples, collected from a publicly owned treatment works (POTW) under normal operating conditions, had little influence on this effectiveness. Higher levels of total suspended solids, such as those associated with normal primary treatment and all treatment stages during storm flow events, appeared to interfere with PMA effectiveness under the sample preparation conditions employed. During normal operating conditions at three different POTWs, greater reductions were observed in PMA-qPCR detectable target sequences of both Enterococcus and Bacteroidales than in total qPCR detectable sequences. These reductions were not as great as those observed for cultivable fecal indicator bacteria in response to wastewater disinfection. Reductions of PMA-qPCR as well as total qPCR detectable target sequences from enterococci and, to a lesser extent, Bacteroidales correlated well with reductions in infectious viruses during both normal and storm flow operating conditions and therefore may have predictive value in determining the efficiency at which these pathogens are removed.

  9. Water quality and sources of fecal coliform bacteria in the Meduxnekeag River, Houlton, Maine

    USGS Publications Warehouse

    Culbertson, Charles W.; Huntington, Thomas G.; Stoeckel, Donald M.; Caldwell, James M.; O'Donnell, Cara

    2014-01-01

    In response to bacterial contamination in the Meduxnekeag River and the desire to manage the watershed to reduce contaminant sources, the Houlton Band of Maliseet Indians (HBMI) and the U.S. Geological Survey began a cooperative effort to establish a baseline of water-quality data that can be used in future studies and to indicate potential sources of nutrient and bacterial contamination. This study was conducted during the summer of 2005 in the Meduxnekeag River Basin near Houlton, Maine. Continuously recorded specific conductance can be a good indicator for water quality. Specific conductance increased downstream from the town of Houlton, between runoff events, and decreased sharply following major runoff events. Collections of discrete samples during the summer of 2005 indicated seasonal positive concentration-discharge relations for total phosphorus and total nitrogen; these results indicate that storm runoff may mobilize and transport these nutrients from the terrestrial environment to the river. Data collected by the HBMI on fecal coliform bacteria indicated that bacterial contamination enters the Meduxnekeag River from multiple paths including tributaries and surface drains (ditches) in developed areas in Houlton, Maine. The Houlton wastewater treatment discharge was not an important source of bacterial contamination. Bacteroidales-based tests for general fecal contamination (Bac32 marker) were predominantly positive in samples that had excessive fecal contamination as indicated by Enterococci density greater than 104 colony-forming units per 100 millilters. Of the 22 samples tested for Bacteroidales-based markers of human-associated fecal contamination (HF134 and HF183), 8 were positive. Of the 22 samples tested for Bacteroidales-based markers of ruminant-associated fecal contamination (CF128 and CF193), 7 were positive. Human fecal contamination was detected consistently at two sites (surface drains in urban areas in the town of Houlton) and occasionally

  10. The Interaction between Heterotrophic Bacteria and Coliform, Fecal Coliform, Fecal Streptococci Bacteria in the Water Supply Networks

    PubMed Central

    AMANIDAZ, Nazak; ZAFARZADEH, Ali; MAHVI, Amir Hossein

    2015-01-01

    Background: This study investigated the interaction between heterotrophic bacteria and coliform, fecal coliforms, fecal streptococci bacteria in water supply networks. Methods: This study was conducted during 2013 on water supply distribution network in Aq Qala City, Golestan Province, Northern Iran and standard methods were applied for microbiological analysis. The surface method was applied to test the heterotrophic bacteria and MPN method was used for coliform, fecal coliform and fecal streptococci bacteria measurements. Results: In 114 samples, heterotrophic bacteria count were over 500 CFU/ml, which the amount of fecal coliform, coliform, and fecal streptococci were 8, 32, and 20 CFU/100 ml, respectively. However, in the other 242 samples, with heterotrophic bacteria count being less than 500 CFU/ml, the amount of fecal coliform, coliform, and fecal streptococci was 7, 23, and 11 CFU/100ml, respectively. The relationship between heterotrophic bacteria, coliforms and fecal streptococci was highly significant (P<0.05). We observed the concentration of coliforms, fecal streptococci bacteria being high, whenever the concentration of heterotrophic bacteria in the water network systems was high. Conclusion: Interaction between heterotrophic bacteria and coliform, fecal coliforms, fecal streptococci bacteria in the Aq Qala City water supply networks was not notable. It can be due to high concentrations of organic carbon, bio-films and nutrients, which are necessary for growth, and survival of all microorganisms. PMID:26811820

  11. The Interaction between Heterotrophic Bacteria and Coliform, Fecal Coliform, Fecal Streptococci Bacteria in the Water Supply Networks.

    PubMed

    Amanidaz, Nazak; Zafarzadeh, Ali; Mahvi, Amir Hossein

    2015-12-01

    This study investigated the interaction between heterotrophic bacteria and coliform, fecal coliforms, fecal streptococci bacteria in water supply networks. This study was conducted during 2013 on water supply distribution network in Aq Qala City, Golestan Province, Northern Iran and standard methods were applied for microbiological analysis. The surface method was applied to test the heterotrophic bacteria and MPN method was used for coliform, fecal coliform and fecal streptococci bacteria measurements. In 114 samples, heterotrophic bacteria count were over 500 CFU/ml, which the amount of fecal coliform, coliform, and fecal streptococci were 8, 32, and 20 CFU/100 ml, respectively. However, in the other 242 samples, with heterotrophic bacteria count being less than 500 CFU/ml, the amount of fecal coliform, coliform, and fecal streptococci was 7, 23, and 11 CFU/100ml, respectively. The relationship between heterotrophic bacteria, coliforms and fecal streptococci was highly significant (P<0.05). We observed the concentration of coliforms, fecal streptococci bacteria being high, whenever the concentration of heterotrophic bacteria in the water network systems was high. Interaction between heterotrophic bacteria and coliform, fecal coliforms, fecal streptococci bacteria in the Aq Qala City water supply networks was not notable. It can be due to high concentrations of organic carbon, bio-films and nutrients, which are necessary for growth, and survival of all microorganisms.

  12. Mortality of fecal bacteria in seawater

    SciTech Connect

    Garcia-Lara, J.; Menon, P.; Servais, P.; Billen, G. )

    1991-03-01

    The authors propose a method for determining the mortality rate for allochthonous bacteria released in aquatic environments without interference due to the loss of culturability in specific culture media. This method consists of following the disappearance of radioactivity from the trichloracetic acid-insoluble fraction in water samples to which ({sup 3}H)thymidine-prelabeled allochthonous bacteria have been added. In coastal seawater, they found that the actual rate of disappearance of fecal bacteria was 1 order of magnitude lower than the rate of loss of culturability on specific media. Minor adaptation of the procedure may facilitate assessment of the effect of protozoan grazing and bacteriophage lysis on the overall bacterial mortality rate.

  13. Distribution of Genetic Marker Concentrations for Fecal Indicator Bacteria in Sewage and Animal Feces

    PubMed Central

    Kelty, Catherine A.; Varma, Manju; Sivaganesan, Mano; Haugland, Richard A.

    2012-01-01

    Very little is known about the density and distribution of fecal indicator bacteria (FIB) genetic markers measured by quantitative real-time PCR (qPCR) in fecal pollution sources. Before qPCR-based FIB technologies can be applied to waste management and public health risk applications, it is vital to characterize the concentrations of these genetic markers in pollution sources (i.e., untreated wastewater and animal feces). We report the distribution of rRNA genetic markers for several general FIB groups, including Clostridium spp., Escherichia coli, enterococci, and Bacteroidales, as determined by qPCR on reference collections consisting of 54 primary influent sewage samples collected from treatment facilities across the United States and fecal samples representing 20 different animal species. Based on raw sewage sample collection data, individual FIB genetic markers exhibited a remarkable similarity in concentration estimates from locations across the United States ranging from Hawaii to Florida. However, there was no significant correlation between genetic markers for most FIB combinations (P > 0.05). In addition, large differences (up to 5 log10 copies) in the abundance of FIB genetic markers were observed between animal species, emphasizing the importance of indicator microorganism selection and animal source contribution for future FIB applications. PMID:22504809

  14. Application of empirical predictive modeling using conventional and alternative fecal indicator bacteria in eastern North Carolina waters

    USGS Publications Warehouse

    Gonzalez, Raul; Conn, Kathleen E.; Crosswell, Joey; Noble, Rachel

    2012-01-01

    Coastal and estuarine waters are the site of intense anthropogenic influence with concomitant use for recreation and seafood harvesting. Therefore, coastal and estuarine water quality has a direct impact on human health. In eastern North Carolina (NC) there are over 240 recreational and 1025 shellfish harvesting water quality monitoring sites that are regularly assessed. Because of the large number of sites, sampling frequency is often only on a weekly basis. This frequency, along with an 18–24 h incubation time for fecal indicator bacteria (FIB) enumeration via culture-based methods, reduces the efficiency of the public notification process. In states like NC where beach monitoring resources are limited but historical data are plentiful, predictive models may offer an improvement for monitoring and notification by providing real-time FIB estimates. In this study, water samples were collected during 12 dry (n = 88) and 13 wet (n = 66) weather events at up to 10 sites. Statistical predictive models for Escherichiacoli (EC), enterococci (ENT), and members of the Bacteroidales group were created and subsequently validated. Our results showed that models for EC and ENT (adjusted R2 were 0.61 and 0.64, respectively) incorporated a range of antecedent rainfall, climate, and environmental variables. The most important variables for EC and ENT models were 5-day antecedent rainfall, dissolved oxygen, and salinity. These models successfully predicted FIB levels over a wide range of conditions with a 3% (EC model) and 9% (ENT model) overall error rate for recreational threshold values and a 0% (EC model) overall error rate for shellfish threshold values. Though modeling of members of the Bacteroidales group had less predictive ability (adjusted R2 were 0.56 and 0.53 for fecal Bacteroides spp. and human Bacteroides spp., respectively), the modeling approach and testing provided information on Bacteroidales ecology. This is the first example of a set of successful statistical

  15. RNA-Based Methods Increase the Detection of Fecal Bacteria and Fecal Identifiers in Environmental Waters

    EPA Science Inventory

    We evaluated the use of qPCR RNA-based methods in the detection of fecal bacteria in environmental waters. We showed that RNA methods can increase the detection of fecal bacteria in multiple water matrices. The data suggest that this is a viable alternative for the detection of a...

  16. RNA-Based Methods Increase the Detection of Fecal Bacteria and Fecal Identifiers in Environmental Waters

    EPA Science Inventory

    We evaluated the use of qPCR RNA-based methods in the detection of fecal bacteria in environmental waters. We showed that RNA methods can increase the detection of fecal bacteria in multiple water matrices. The data suggest that this is a viable alternative for the detection of a...

  17. Fecal bacteria source characterization and sensitivity analysis of SWAT 2005

    USDA-ARS?s Scientific Manuscript database

    The Soil and Water Assessment Tool (SWAT) version 2005 includes a microbial sub-model to simulate fecal bacteria transport at the watershed scale. The objectives of this study were to demonstrate methods to characterize fecal coliform bacteria (FCB) source loads and to assess the model sensitivity t...

  18. Characterizing relationships among fecal indicator bacteria ...

    EPA Pesticide Factsheets

    Bed sediments of streams and rivers may store high concentrations of fecal indicator bacteria (FIB) and pathogens. Due to resuspension events, these contaminants can be mobilized into the water column and affect overall water quality. Other bacterial indicators such as microbial source tracking (MST) markers, developed to determine potential sources of fecal contamination, can also be resuspended from bed sediments. The primary objective of this study was to predict occurrence of waterborne pathogens in water and streambed sediments using a simple statistical model that includes traditionally measured FIB, environmental parameters and source allocation, using MST markers as predictor variables. Synoptic sampling events were conducted during baseflow conditions downstream from agricultural (AG), forested (FORS), and wastewater pollution control plant (WPCP) land uses. Concentrations of FIB and MST markers were measured in water and sediments, along with occurrences of the enteric pathogens Campylobacter, Listeria and Salmonella, and the virulence gene that carries Shiga toxin, stx2. Pathogens were detected in water more often than in underlying sediments. Shiga toxin was significantly related to land use, with concentrations of the ruminant marker selected as an independent variable that could correctly classify 76% and 64% of observed Shiga toxin occurrences in water and sediment, respectively. FIB concentrations and water quality parameters were also selected a

  19. Characterizing relationships among fecal indicator bacteria ...

    EPA Pesticide Factsheets

    Bed sediments of streams and rivers may store high concentrations of fecal indicator bacteria (FIB) and pathogens. Due to resuspension events, these contaminants can be mobilized into the water column and affect overall water quality. Other bacterial indicators such as microbial source tracking (MST) markers, developed to determine potential sources of fecal contamination, can also be resuspended from bed sediments. The primary objective of this study was to predict occurrence of waterborne pathogens in water and streambed sediments using a simple statistical model that includes traditionally measured FIB, environmental parameters and source allocation, using MST markers as predictor variables. Synoptic sampling events were conducted during baseflow conditions downstream from agricultural (AG), forested (FORS), and wastewater pollution control plant (WPCP) land uses. Concentrations of FIB and MST markers were measured in water and sediments, along with occurrences of the enteric pathogens Campylobacter, Listeria and Salmonella, and the virulence gene that carries Shiga toxin, stx2. Pathogens were detected in water more often than in underlying sediments. Shiga toxin was significantly related to land use, with concentrations of the ruminant marker selected as an independent variable that could correctly classify 76% and 64% of observed Shiga toxin occurrences in water and sediment, respectively. FIB concentrations and water quality parameters were also selected a

  20. Bacteroidales Diversity in Ring-Billed Gulls (Laurus delawarensis) Residing at Lake Michigan Beaches▿

    PubMed Central

    Jeter, Sonja N.; McDermott, Colleen M.; Bower, Patricia A.; Kinzelman, Julie L.; Bootsma, Melinda J.; Goetz, Giles W.; McLellan, Sandra L.

    2009-01-01

    This study investigated the occurrence and diversity of Bacteroidales fecal bacteria in gulls residing in the Great Lakes region. Members of this bacterial order have been widely employed as human and bovine host-specific markers of fecal pollution; however, few studies have focused on gulls, which can be a major source of fecal indicator bacteria and pathogens at beaches. We found a low but consistent occurrence of Bacteroidales in gulls at five beaches in three different counties spanning the Wisconsin shoreline of Lake Michigan. The percentages of gulls positive for Bacteroidales were 4 to 8% at beaches in the southern part of the state and 8 to 50% at beaches in the north. Sequencing of 931 clones from seven gull Bacteroidales 16S rRNA gene libraries revealed a large amount of diversity in both individual and pooled gull fecal samples. Two libraries constructed from pooled gull fecal samples (n = 5 and n = 6) did not have a greater richness of sequences than individual samples, suggesting that even within a single gull diversity is high and an extensive sequencing effort is needed to characterize the populations. Estimates of the numbers of operational taxonomic units (OTUs) for the libraries obtained using different similarity levels revealed a large amount of microdiveristy with a limited number of OTUs at the 95% similarity level. Gull sequences were clustered by the beach from which they were collected, suggesting that there were geographic effects on the distribution of Bacteriodales. More than 53% of the 16S rRNA gene sequences from gulls at the southern beaches were associated with the family Porphyromonadaceae, primarily the genus Parabacteroides, whereas sequences from gulls at the northern beaches were comprised of Bacteroidaceae and Prevotellaceae sequences. Comparison of gull sequences with sequences from goose, canine, raccoon, and sewage sources revealed distinct clusters of closely related gull sequences; however, these sequences were widely

  1. Bacteroidales diversity in ring-billed gulls (Laurus delawarensis) residing at Lake Michigan beaches.

    PubMed

    Jeter, Sonja N; McDermott, Colleen M; Bower, Patricia A; Kinzelman, Julie L; Bootsma, Melinda J; Goetz, Giles W; McLellan, Sandra L

    2009-03-01

    This study investigated the occurrence and diversity of Bacteroidales fecal bacteria in gulls residing in the Great Lakes region. Members of this bacterial order have been widely employed as human and bovine host-specific markers of fecal pollution; however, few studies have focused on gulls, which can be a major source of fecal indicator bacteria and pathogens at beaches. We found a low but consistent occurrence of Bacteroidales in gulls at five beaches in three different counties spanning the Wisconsin shoreline of Lake Michigan. The percentages of gulls positive for Bacteroidales were 4 to 8% at beaches in the southern part of the state and 8 to 50% at beaches in the north. Sequencing of 931 clones from seven gull Bacteroidales 16S rRNA gene libraries revealed a large amount of diversity in both individual and pooled gull fecal samples. Two libraries constructed from pooled gull fecal samples (n = 5 and n = 6) did not have a greater richness of sequences than individual samples, suggesting that even within a single gull diversity is high and an extensive sequencing effort is needed to characterize the populations. Estimates of the numbers of operational taxonomic units (OTUs) for the libraries obtained using different similarity levels revealed a large amount of microdiveristy with a limited number of OTUs at the 95% similarity level. Gull sequences were clustered by the beach from which they were collected, suggesting that there were geographic effects on the distribution of Bacteriodales. More than 53% of the 16S rRNA gene sequences from gulls at the southern beaches were associated with the family Porphyromonadaceae, primarily the genus Parabacteroides, whereas sequences from gulls at the northern beaches were comprised of Bacteroidaceae and Prevotellaceae sequences. Comparison of gull sequences with sequences from goose, canine, raccoon, and sewage sources revealed distinct clusters of closely related gull sequences; however, these sequences were widely

  2. Fecal Indicator Bacteria and Environmental Observations: Validation of Virtual Beach

    EPA Science Inventory

    Contamination of recreational waters by fecal material is often assessed using indicator bacteria such as enterococci. Enumeration based on culturing methods can take up to 48 hours to complete, limiting the accuracy of water quality evaluations. Molecular microbial techniques em...

  3. Fecal Indicator Bacteria and Environmental Observations: Validation of Virtual Beach

    EPA Science Inventory

    Contamination of recreational waters by fecal material is often assessed using indicator bacteria such as enterococci. Enumeration based on culturing methods can take up to 48 hours to complete, limiting the accuracy of water quality evaluations. Molecular microbial techniques em...

  4. Sources of fecal indicator bacteria to groundwater, Malibu Lagoon and the near-shore ocean, Malibu, California, USA

    USGS Publications Warehouse

    Izbicki, John A.; Swarzenski, Peter W.; Burton, Carmen A.; Van De Werfhorst, Laurie; Holden, Patricia A.; Dubinsky, Eric A.

    2012-01-01

    Onsite wastewater treatment systems (OWTS) used to treat residential and commercial sewage near Malibu, California have been implicated as a possible source of fecal indicator bacteria (FIB) to Malibu Lagoon and the near-shore ocean. For this to occur, treated wastewater must first move through groundwater before discharging to the Lagoon or ocean. In July 2009 and April 2010, δ18O and δD data showed that some samples from water-table wells contained as much as 70% wastewater; at that time FIB concentrations in those samples were generally less than the detection limit of 1 Most Probable Number (MPN) per 100 milliliters (mL). In contrast, Malibu Lagoon had total coliform, Escherichia coli, and enterococci concentrations as high as 650,000, 130,000, and 5,500 MPN per 100 mL, respectively, and as many as 12% of samples from nearby ocean beaches exceeded the U.S. Environmental Protection Agency single sample enterococci standard for marine recreational water of 104 MPN per 100 mL. Human-associated Bacteroidales, an indicator of human-fecal contamination, were not detected in water from wells, Malibu Lagoon, or the near-shore ocean. Similarly, microarray (PhyloChip) data show Bacteroidales and Fimicutes Operational Taxanomic Units (OTUs) present in OWTS were largely absent in groundwater; in contrast, 50% of Bacteroidales and Fimicutes OTUs present in the near-shore ocean were also present in gull feces. Terminal-Restriction Length Fragment Polymorphism (T-RFLP) and phospholipid fatty acid (PLFA) data showed that microbial communities in groundwater were different and less abundant than communities in OWTS, Malibu Lagoon, or the near-shore ocean. However, organic compounds indicative of wastewater (such as fecal sterols, bisphenol-A and cosmetics) were present in groundwater having a high percentage of wastewater and were present in groundwater discharging to the ocean. FIB in the near-shore ocean varied with tides, ocean swells, and waves. Movement of water from

  5. Survival and persistence of host-associated Bacteroidales cells and DNA in comparison with Escherichia coli and Enterococcus in freshwater sediments as quantified by PMA-qPCR and qPCR.

    PubMed

    Kim, Minji; Wuertz, Stefan

    2015-12-15

    Decay of the fecal source identifier Bacteroidales in sediments has not been studied until now. Two types of microcosms inoculated with human, cow and dog feces were constructed to investigate the survival and persistence of host-associated Bacteroidales cells and their DNA, respectively, in freshwater sediments: (i) a completely anaerobic microcosm where feces were entirely mixed with sediments for estimating decay of Bacteroidales in oxygen-free sediments at two temperatures (6 °C and 20 °C) and (ii) a core microcosm where feces in the overlying water column settled on top of undisturbed core sediments. Quantitative PCR (qPCR) along with propidium monoazide (PMA) was used to differentiate between genetic markers present in intact cells and total intracellular as well as extracellular marker DNA. Regulated fecal indicator bacteria were measured by cultivation (Escherichia coli and Enterococcus) and qPCR (Enterococcus) in relation to Bacteroidales-associated host markers. In anaerobic microcosms, the survival and persistence of Bacteroidales cells and DNA in sediments were considerably extended, especially at the lower temperature of 6 °C, with two-log reduction times (T99) >56 d (cells) and >169 d (DNA). Bacteroidales DNA persisted up to five times longer than cells in anaerobic microcosms at 6 °C, whereas decay rates of cells and DNA were not significantly different at 20 °C in anaerobic microcosms. In core microcosms, the levels of Bacteroidales cells and DNA decreased approximately six times more slowly in sediments than in overlying water; T99 values of Bacteroidales cells and DNA were 6-9 d (water) and 29-82 d (sediment). The survival of universal, human-, ruminant- and dog-associated Bacteroidales cells in sediments was similar in both microcosms under each given condition, as was the persistence of DNA. Decay rate constants of Bacteroidales cells and DNA were comparable with those of cultivable Enterococcus and E. coli cells in core sediments while

  6. Interlaboratory comparison of real-time pcr protocols for quantification of general fecal indicator bacteria

    USGS Publications Warehouse

    Shanks, O.C.; Sivaganesan, M.; Peed, L.; Kelty, C.A.; Blackwood, A.D.; Greene, M.R.; Noble, R.T.; Bushon, R.N.; Stelzer, E.A.; Kinzelman, J.; Anan'Eva, T.; Sinigalliano, C.; Wanless, D.; Griffith, J.; Cao, Y.; Weisberg, S.; Harwood, V.J.; Staley, C.; Oshima, K.H.; Varma, M.; Haugland, R.A.

    2012-01-01

    The application of quantitative real-time PCR (qPCR) technologies for the rapid identification of fecal bacteria in environmental waters is being considered for use as a national water quality metric in the United States. The transition from research tool to a standardized protocol requires information on the reproducibility and sources of variation associated with qPCR methodology across laboratories. This study examines interlaboratory variability in the measurement of enterococci and Bacteroidales concentrations from standardized, spiked, and environmental sources of DNA using the Entero1a and GenBac3 qPCR methods, respectively. Comparisons are based on data generated from eight different research facilities. Special attention was placed on the influence of the DNA isolation step and effect of simplex and multiplex amplification approaches on interlaboratory variability. Results suggest that a crude lysate is sufficient for DNA isolation unless environmental samples contain substances that can inhibit qPCR amplification. No appreciable difference was observed between simplex and multiplex amplification approaches. Overall, interlaboratory variability levels remained low (<10% coefficient of variation) regardless of qPCR protocol. ?? 2011 American Chemical Society.

  7. Interlaboratory comparison of real-time PCR protocols for quantification of general fecal indicator bacteria.

    PubMed

    Shanks, Orin C; Sivaganesan, Mano; Peed, Lindsay; Kelty, Catherine A; Blackwood, A Denene; Greene, Monica R; Noble, Rachel T; Bushon, Rebecca N; Stelzer, Erin A; Kinzelman, Julie; Anan'eva, Tamara; Sinigalliano, Christopher; Wanless, David; Griffith, John; Cao, Yiping; Weisberg, Steve; Harwood, Valarie J; Staley, Christopher; Oshima, Kevin H; Varma, Manju; Haugland, Richard A

    2012-01-17

    The application of quantitative real-time PCR (qPCR) technologies for the rapid identification of fecal bacteria in environmental waters is being considered for use as a national water quality metric in the United States. The transition from research tool to a standardized protocol requires information on the reproducibility and sources of variation associated with qPCR methodology across laboratories. This study examines interlaboratory variability in the measurement of enterococci and Bacteroidales concentrations from standardized, spiked, and environmental sources of DNA using the Entero1a and GenBac3 qPCR methods, respectively. Comparisons are based on data generated from eight different research facilities. Special attention was placed on the influence of the DNA isolation step and effect of simplex and multiplex amplification approaches on interlaboratory variability. Results suggest that a crude lysate is sufficient for DNA isolation unless environmental samples contain substances that can inhibit qPCR amplification. No appreciable difference was observed between simplex and multiplex amplification approaches. Overall, interlaboratory variability levels remained low (<10% coefficient of variation) regardless of qPCR protocol.

  8. Chapter A7. Section 7.1. Fecal Indicator Bacteria

    USGS Publications Warehouse

    Myers, Donna N.; Sylvester, Marc A.

    1997-01-01

    Fecal indicator bacteria are used to assess the microbiological quality of water because, although not typically disease causing, they are correlated with the presence of several waterborne disease-causing organisms. The concentration of indicator bacteria is a measure of water safety for body-contact recreation or for consumption. This report provides information on the equipment, sampling protocols, and identification, enumeration, and calculation procedures that are in standard use by U.S. Geological Survey (USGS) personnel for the collection of data on fecal indicator bacteria.

  9. Application of empirical predictive modeling using conventional and alternative fecal indicator bacteria in eastern North Carolina waters.

    PubMed

    Gonzalez, Raul A; Conn, Kathleen E; Crosswell, Joseph R; Noble, Rachel T

    2012-11-15

    Coastal and estuarine waters are the site of intense anthropogenic influence with concomitant use for recreation and seafood harvesting. Therefore, coastal and estuarine water quality has a direct impact on human health. In eastern North Carolina (NC) there are over 240 recreational and 1025 shellfish harvesting water quality monitoring sites that are regularly assessed. Because of the large number of sites, sampling frequency is often only on a weekly basis. This frequency, along with an 18-24 h incubation time for fecal indicator bacteria (FIB) enumeration via culture-based methods, reduces the efficiency of the public notification process. In states like NC where beach monitoring resources are limited but historical data are plentiful, predictive models may offer an improvement for monitoring and notification by providing real-time FIB estimates. In this study, water samples were collected during 12 dry (n = 88) and 13 wet (n = 66) weather events at up to 10 sites. Statistical predictive models for Escherichiacoli (EC), enterococci (ENT), and members of the Bacteroidales group were created and subsequently validated. Our results showed that models for EC and ENT (adjusted R(2) were 0.61 and 0.64, respectively) incorporated a range of antecedent rainfall, climate, and environmental variables. The most important variables for EC and ENT models were 5-day antecedent rainfall, dissolved oxygen, and salinity. These models successfully predicted FIB levels over a wide range of conditions with a 3% (EC model) and 9% (ENT model) overall error rate for recreational threshold values and a 0% (EC model) overall error rate for shellfish threshold values. Though modeling of members of the Bacteroidales group had less predictive ability (adjusted R(2) were 0.56 and 0.53 for fecal Bacteroides spp. and human Bacteroides spp., respectively), the modeling approach and testing provided information on Bacteroidales ecology. This is the first example of a set of successful

  10. Fecal indicator bacteria variability in samples pumped from monitoring wells.

    PubMed

    Kozuskanich, J; Novakowski, K S; Anderson, B C

    2011-01-01

    The detection of microbiological contamination in drinking water from groundwater wells is often made with a limited number of samples that are collected using traditional geochemical sampling protocols. The objective of this study is to examine the variability of fecal indicator bacteria, as observed using discrete samples, due to pumping. Two wells were instrumented as multilevel piezometers in a bedrock aquifer, and bacterial enumeration was conducted on a total of 166 samples (for total coliform, fecal coliform, Escherichia coli, and fecal streptococci) using standard membrane filtration methods. Five tests were conducted using pumping rates ranging from 0.3 to 17 L/min in a variety of purging scenarios, which included constant and variable (incremental increase and decrease) flow. The results clearly show a rapid and reproducible, 1 to 2 log-unit decrease in fecal indicator bacteria at the onset of pumping to stabilized, low-level concentrations prior to the removal of three to five well volumes. The pumping rate was not found to be correlated with the magnitude of observed bacterial counts. Based on the results, we suggest sampling protocols for fecal indicator bacteria that include multiple collections during the course of pumping, including early-time samples, and consider other techniques such as microscopic enumeration when assessing the source of bacteria from the well-aquifer system.

  11. Fecal-coliform bacteria in extended-aeration plant sludge

    SciTech Connect

    Anderson, M.; Kester, G.; Arant, S.

    1998-07-01

    The concentration of fecal-coliform bacteria in sludge from extended-aeration plants was analyzed for compliance with new state and federal land application requirements. This study was initiated to determine if additional digestion would be necessary for plants to meet the new pathogen standards of less than 2 million CFU per gm of solids. Sludge was found to contain less than 2 million fecal coliform bacteria/gm of sludge as a result of a combination or aerobic digestion and/or long term storage.

  12. Profiling Living Bacteria Informs Preparation of Fecal Microbiota Transplantations

    PubMed Central

    Chu, Nathaniel D.; Smith, Mark B.; Perrotta, Allison R.; Kassam, Zain; Alm, Eric J.

    2017-01-01

    Fecal microbiota transplantation is a compelling treatment for recurrent Clostridium difficile infections, with potential applications against other diseases associated with changes in gut microbiota. But variability in fecal bacterial communities—believed to be the therapeutic agent—can complicate or undermine treatment efficacy. To understand the effects of transplant preparation methods on living fecal microbial communities, we applied a DNA-sequencing method (PMA-seq) that uses propidium monoazide (PMA) to differentiate between living and dead fecal microbes, and we created an analysis pipeline to identify individual bacteria that change in abundance between samples. We found that oxygen exposure degraded fecal bacterial communities, whereas freeze-thaw cycles and lag time between donor defecation and transplant preparation had much smaller effects. Notably, the abundance of Faecalibacterium prausnitzii—an anti-inflammatory commensal bacterium whose absence is linked to inflammatory bowel disease—decreased with oxygen exposure. Our results indicate that some current practices for preparing microbiota transplant material adversely affect living fecal microbial content and highlight PMA-seq as a valuable tool to inform best practices and evaluate the suitability of clinical fecal material. PMID:28125667

  13. Profiling Living Bacteria Informs Preparation of Fecal Microbiota Transplantations.

    PubMed

    Chu, Nathaniel D; Smith, Mark B; Perrotta, Allison R; Kassam, Zain; Alm, Eric J

    2017-01-01

    Fecal microbiota transplantation is a compelling treatment for recurrent Clostridium difficile infections, with potential applications against other diseases associated with changes in gut microbiota. But variability in fecal bacterial communities-believed to be the therapeutic agent-can complicate or undermine treatment efficacy. To understand the effects of transplant preparation methods on living fecal microbial communities, we applied a DNA-sequencing method (PMA-seq) that uses propidium monoazide (PMA) to differentiate between living and dead fecal microbes, and we created an analysis pipeline to identify individual bacteria that change in abundance between samples. We found that oxygen exposure degraded fecal bacterial communities, whereas freeze-thaw cycles and lag time between donor defecation and transplant preparation had much smaller effects. Notably, the abundance of Faecalibacterium prausnitzii-an anti-inflammatory commensal bacterium whose absence is linked to inflammatory bowel disease-decreased with oxygen exposure. Our results indicate that some current practices for preparing microbiota transplant material adversely affect living fecal microbial content and highlight PMA-seq as a valuable tool to inform best practices and evaluate the suitability of clinical fecal material.

  14. Growth of infant fecal bacteria on commercial prebiotics.

    PubMed

    Bunešová, Věra; Vlková, Eva; Rada, Vojtěch; Kňazovická, Vladimíra; Ročková, Sárka; Geigerová, Martina; Božik, Matěj

    2012-07-01

    Fecal bacteria from 33 infants (aged 1 to 6 months) were tested for growth on commercial prebiotics. The children were born vaginally (20) or by caesarean section (13). Bifidobacteria, lactobacilli, gram-negative bacteria, Escherichia coli, and total anaerobes in fecal samples were enumerated by selective agars and fluorescence in situ hybridization. The total fecal bacteria were inoculated into cultivation media containing 2 % Vivinal® (galactooligosaccharides-GOS) or Raftilose® P95 (fructooligosaccharides-FOS) as a single carbon source and bacteria were enumerated again after 24 h of anaerobic cultivation. Bifidobacteria dominated, reaching counts of 9-10 log colony-forming units (CFU)/g in 17 children born vaginally and in seven children delivered by caesarean section. In these infants, lactobacilli were more frequently detected and a lower number of E. coli and gram-negative bacteria were determined compared to bifidobacteria-negative infants. Clostridia dominated in children without bifidobacteria, reaching counts from 7 to 9 log CFU/g. Both prebiotics supported all groups of bacteria tested. In children with naturally high counts of bifidobacteria, bifidobacteria dominated also after cultivation on prebiotics, reaching counts from 8.23 to 8.77 log CFU/mL. In bifidobacteria-negative samples, clostridia were supported by prebiotics, reaching counts from 7.17 to 7.69 log CFU/mL. There were no significant differences between bacterial growth on Vivinal® and Raftilose® P95 and counts determined by cultivation and FISH. Prebiotics should selectively stimulate the growth of desirable bacteria such as bifidobacteria and lactobacilli. However, our results showed that commercially available FOS and GOS may stimulate also other fecal bacteria.

  15. Transport and attenuation of Salmonella enterica, fecal indicator bacteria and a poultry litter marker gene are correlated in soil columns.

    PubMed

    Mantha, Sirisha; Anderson, Angela; Acharya, Saraswati Poudel; Harwood, Valerie J; Weidhaas, Jennifer

    2017-11-15

    Millions of tons of fecal-contaminated poultry litter are applied to U.S. agricultural fields annually. Precipitation and irrigation facilitate transport of fecal-derived pathogens and fecal indicator bacteria (FIB) to groundwater. The goal of this study was to compare transport of pathogens, FIB, and a microbial source tracking marker gene for poultry litter (LA35) in a simulated soil-to-groundwater system. Nine laboratory soil columns containing four different soil types were used to evaluate microbial transport to groundwater via infiltration. Quantitative polymerase chain reaction was used to monitor Salmonella enterica Typhimurium, Escherichia coli, Enterococcus spp., Brevibacterium sp. LA35 and Bacteroidales leached from soil columns inoculated with poultry litter. S. enterica was correlated with LA35 poultry litter marker gene and FIB concentrations in column soils containing organic matter, but not in acid washed sands. In contrast, S. enterica was found to correlate with LA35 and FIB in the leachate from columns containing sand, but not with leachate from organic soil columns. The majority of recovered DNA was found in leachate of predominately sandy soil columns, and in the soil of loamy columns. At least 90% of the DNA retained in soils for each microbial target was found in the top 3cm of the column. These studies suggest that poultry litter associated pathogens and FIB are rapidly released from litter, but are influenced by complex attenuation mechanisms during infiltration, including soil type. This study advances our understanding of the potential for subsurface transport of poultry litter associated pathogens and FIB, and support the use of the LA35 marker gene for evaluating poultry litter impacts on groundwater. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Diversity, abundance, and possible sources of fecal bacteria in the Yangtze River.

    PubMed

    Sun, Haohao; He, Xiwei; Ye, Lin; Zhang, Xu-Xiang; Wu, Bing; Ren, Hongqiang

    2017-03-01

    The fecal bacteria in natural waters may pose serious risks on human health. Although many source tracking methods have been developed and used to determine the possible sources of the fecal pollution, little is known about the overall diversity and abundance of fecal bacterial community in natural waters. In this study, a method based on fecal bacterial sequence library was introduced to evaluate the fecal bacterial profile in the Yangtze River (Nanjing section). Our results suggested that the Yangtze River water harbors diverse fecal bacteria. Fifty-eight fecal operational taxonomic units (97% identity level) were detected in the Yangtze River water samples and the relative abundance of fecal bacteria in these samples ranged from 0.1 to 8%. It was also found that the relative abundances of the fecal bacteria in locations near to the downstream of wastewater treatment plants were obviously higher than those in other locations. However, the high abundance of fecal bacteria could decrease to the normal level in 2~4 km in the river due to degradation or dilution, and the overall fecal bacteria level changed little when the Yangtze River flew through the Nanjing City. Moreover, the fecal bacteria in the Yangtze River water were found to be highly associated (Spearman rho = 0.804, P < 0.001) with the potential pathogenic bacteria. Collectively, the findings in this study reveal the diversity, abundance, and possible sources of fecal bacteria in the Yangtze River and advance our understandings of the fecal bacteria community in the natural waters.

  17. Correlative assessment of fecal indicators using human mitochondrial DNA as a direct marker.

    PubMed

    Kapoor, Vikram; Smith, Christopher; Santo Domingo, Jorge W; Lu, Ting; Wendell, David

    2013-09-17

    Identifying the source of surface water fecal contamination is paramount to mitigating pollution and risk to human health. Fecal bacteria such as E. coli have been staple indicator organisms for over a century, however there remains uncertainty with E. coli-based metrics since these bacteria are abundant in the environment. The relationships between the presence of direct indicator of human waste (human mitochondrial DNA), human-specific Bacteroidales, and E. coli were studied for water samples taken from an urban creek system (Duck Creek Watershed, Cincinnati, OH) impacted by combined sewer overflows. Logistic regression analysis shows that human-specific Bacteroidales correlates much more closely to human mitochondrial DNA (R = 0.62) relative to E. coli (R = 0.33). We also examine the speciation of Bacteroidales within the Duck Creek Watershed using next-generation sequencing technology (Ion Torrent) and show the most numerous populations to be associated with sewage. Here we demonstrate that human-specific Bacteroidales closely follow the dynamics of human mitochondrial DNA concentration changes, indicating that these obligate anaerobes are more accurate than E. coli for fecal source tracking, lending further support to risk overestimation using coliforms, especially fecal coliforms and E. coli.

  18. Addressing uncertainty in fecal indicator bacteria dark inactivation rates.

    PubMed

    Gronewold, Andrew D; Myers, Luke; Swall, Jenise L; Noble, Rachel T

    2011-01-01

    Assessing the potential threat of fecal contamination in surface water often depends on model forecasts which assume that fecal indicator bacteria (FIB, a proxy for the concentration of pathogens found in fecal contamination from warm-blooded animals) are lost or removed from the water column at a certain rate (often referred to as an "inactivation" rate). In efforts to reduce human health risks in these water bodies, regulators enforce limits on easily-measured FIB concentrations, commonly reported as most probable number (MPN) and colony forming unit (CFU) values. Accurate assessment of the potential threat of fecal contamination, therefore, depends on propagating uncertainty surrounding "true" FIB concentrations into MPN and CFU values, inactivation rates, model forecasts, and management decisions. Here, we explore how empirical relationships between FIB inactivation rates and extrinsic factors might vary depending on how uncertainty in MPN values is expressed. Using water samples collected from the Neuse River Estuary (NRE) in eastern North Carolina, we compare Escherichia coli (EC) and Enterococcus (ENT) dark inactivation rates derived from two statistical models of first-order loss; a conventional model employing ordinary least-squares (OLS) regression with MPN values, and a novel Bayesian model utilizing the pattern of positive wells in an IDEXX Quanti-Tray®/2000 test. While our results suggest that EC dark inactivation rates tend to decrease as initial EC concentrations decrease and that ENT dark inactivation rates are relatively consistent across different ENT concentrations, we find these relationships depend upon model selection and model calibration procedures. We also find that our proposed Bayesian model provides a more defensible approach to quantifying uncertainty in microbiological assessments of water quality than the conventional MPN-based model, and that our proposed model represents a new strategy for developing robust relationships between

  19. Evaluating spatial and temporal variability of fecal coliform bacteria loads at Pelahatchie Watershed in Mississippi

    USDA-ARS?s Scientific Manuscript database

    Bacterial contaminations of surface waters are an increasing concern for scientists because pathogenic bacteria can cause adverse effects on human health. This research was performed to investigate spatial and seasonal variability of fecal coliform bacteria (FCB) concentrations from the Pelahatchie ...

  20. Significance of beach geomorphology on fecal indicator bacteria levels.

    PubMed

    Donahue, Allison; Feng, Zhixuan; Kelly, Elizabeth; Reniers, Ad; Solo-Gabriele, Helena M

    2017-08-15

    Large databases of fecal indicator bacteria (FIB) measurements are available for coastal waters. With the assistance of satellite imagery, we illustrated the power of assessing data for many sites by evaluating beach features such as geomorphology, distance from rivers and canals, presence of piers and causeways, and degree of urbanization coupled with the enterococci FIB database for the state of Florida. We found that beach geomorphology was the primary characteristic associated with enterococci levels that exceeded regulatory guidelines. Beaches in close proximity to marshes or within bays had higher enterococci exceedances in comparison to open coast beaches. For open coast beaches, greater enterococci exceedances were associated with nearby rivers and higher levels of urbanization. Piers and causeways had a minimal contribution, as their effect was often overwhelmed by beach geomorphology. Results can be used to understand the potential causes of elevated enterococci levels and to promote public health. Copyright © 2017 Elsevier Ltd. All rights reserved.

  1. Concentrations of host-specific and generic fecal markers measured by quantitative PCR in raw sewage and fresh animal feces.

    PubMed

    Silkie, Sarah S; Nelson, Kara L

    2009-11-01

    We measured the concentrations of four host-specific (human, dog, cow, and horse Bacteroidales), four generic fecal (16S total Bacteroidales and Escherichia coli, 23S Enterococcus and uidA E. coli,) and two universal bacterial (16S universal and rpoB universal) DNA targets by qPCR in raw sewage and pooled fecal samples from dogs, cows, horses, and Canada Geese. A spiking protocol using the non-fecal bacterium Pseudomonas syringae pph 6 was developed to estimate the recovery of DNA from fecal and environmental samples. The measured fecal marker concentrations were used to calculate baseline ratios and variability of host-specific to generic indicators for each host type. The host-specific markers were found in high concentrations (8-9 log(10)copies/g dry wt.) in their respective hosts' samples, which were equal to or greater than the concentrations of generic E. coli and Enterococcus markers, lending support to the use of host-specific and generic Bacteroidales as sensitive indicators of fecal pollution. The host-specific markers formed a consistent percentage of total Bacteroidales in target host feces and raw sewage, with human-specific comprising 82%, dog-specific 6%, cow-specific 4% and horse-specific 2%. Based on this limited data set, the measurement of host-specific indicators by qPCR has several promising applications. These applications include determining the percentage of total Bacteroidales contributed by a specific host type, using the ratios of host-specific markers to E. coli or Enterococcus to estimate the contribution of each source to these regulated fecal indicator bacteria, and estimating the mass of feces from each host type in environmental samples.

  2. Stimulation of fecal bacteria in ambient waters by experimental inputs of organic and inorganic phosphorus.

    PubMed

    Chudoba, Elizabeth A; Mallin, Michael A; Cahoon, Lawrence B; Skrabal, Stephen A

    2013-06-15

    Fecal microbial pollution of recreational and shellfishing waters is a major human health and economic issue. Microbial pollution sourced from stormwater runoff is especially widespread, and strongly associated with urbanization. However, non-point source nutrient pollution is also problematic, and may come from sources different from fecal-derived pollution (i.e. fertilization of farm fields, lawns and gardens, and ornamental urban areas). Fecal bacteria require nutrients; thus the impact of such nutrient loading on survival and abundance of fecal coliform bacteria in ambient waters was experimentally investigated in a constructed wetland in coastal North Carolina, USA. A series of nutrient-addition bioassays testing impacts of inorganic and organic nitrogen and phosphorus demonstrated that additions of neither organic nor inorganic nitrogen stimulated fecal coliform bacteria. However, phosphorus additions provided significant stimulation of fecal coliform growth at times; on other occasions such additions did not. Dilution bioassays combined with nutrient additions were subsequently devised to assess potential impacts of microzooplankton grazing on the target fecal bacteria populations. Results demonstrated grazing to be a significant bacterial reduction factor in 63% of tests, potentially obscuring nutrient effects. Thus, combining dilution experiments with nutrient addition bioassays yielded simultaneous information on microzooplankton grazing rates on fecal bacteria, fecal bacterial growth rates, and nutrient limitation. Overall, when tested against a non-amended control, additions of either organic or inorganic phosphorus significantly stimulated fecal coliform bacterial growth on 50% of occasions tested, with organic phosphorus generally providing greater stimulation. The finding of significant phosphorus stimulation of fecal bacteria indicates that extraneous nutrient loading can, at times, augment the impacts of fecal microbial pollution of shellfishing

  3. Addressing Uncertainty in Fecal Indicator Bacteria Dark Inactivation Rates

    EPA Science Inventory

    Fecal contamination is a leading cause of surface water quality degradation. Roughly 20% of all total maximum daily load assessments approved by the United States Environmental Protection Agency since 1995, for example, address water bodies with unacceptably high fecal indicator...

  4. Addressing Uncertainty in Fecal Indicator Bacteria Dark Inactivation Rates

    EPA Science Inventory

    Fecal contamination is a leading cause of surface water quality degradation. Roughly 20% of all total maximum daily load assessments approved by the United States Environmental Protection Agency since 1995, for example, address water bodies with unacceptably high fecal indicator...

  5. Implications of Fecal Bacteria Input from Latrine-Polluted Ponds for Wells in Sandy Aquifers

    PubMed Central

    Knappett, Peter S. K.; McKay, Larry D.; Layton, Alice; Williams, Daniel E.; Alam, Md. J.; Huq, Md. R.; Mey, Jacob; Feighery, John E.; Culligan, Patricia J.; Mailloux, Brian J.; Zhuang, Jie; Escamilla, Veronica; Emch, Michael; Perfect, Edmund; Sayler, Gary S.; Ahmed, Kazi M.; van Geen, Alexander

    2012-01-01

    Ponds receiving latrine effluents may serve as sources of fecal contamination to shallow aquifers tapped by millions of tube-wells in Bangladesh. To test this hypothesis, transects of monitoring wells radiating away from four ponds were installed in a shallow sandy aquifer underlying a densely populated village and monitored for 14 months. Two of the ponds extended to medium sand. Another pond was sited within silty sand and the last in silt. The fecal indicator bacterium E. coli was rarely detected along the transects during the dry season and was only detected near the ponds extending to medium sand up to 7 m away during the monsoon. A log-linear decline in E. coli and Bacteroidales concentrations with distance along the transects in the early monsoon indicates that ponds excavated in medium sand were the likely source of contamination. Spatial removal rates ranged from 0.5-1.3 log10/m. After the ponds were artificially filled with groundwater to simulate the impact of a rain storm, E. coli levels increased near a pond recently excavated in medium sand, but no others. These observations show that adjacent sediment grain-size and how recently a pond was excavated influence how much fecal contamination ponds receiving latrine effluents contribute to neighboring groundwater. PMID:22191430

  6. Implications of fecal bacteria input from latrine-polluted ponds for wells in sandy aquifers.

    PubMed

    Knappett, Peter S K; McKay, Larry D; Layton, Alice; Williams, Daniel E; Alam, Md J; Huq, Md R; Mey, Jacob; Feighery, John E; Culligan, Patricia J; Mailloux, Brian J; Zhuang, Jie; Escamilla, Veronica; Emch, Michael; Perfect, Edmund; Sayler, Gary S; Ahmed, Kazi M; van Geen, Alexander

    2012-02-07

    Ponds receiving latrine effluents may serve as sources of fecal contamination to shallow aquifers tapped by millions of tube-wells in Bangladesh. To test this hypothesis, transects of monitoring wells radiating away from four ponds were installed in a shallow sandy aquifer underlying a densely populated village and monitored for 14 months. Two of the ponds extended to medium sand. Another pond was sited within silty sand and the last in silt. The fecal indicator bacterium E. coli was rarely detected along the transects during the dry season and was only detected near the ponds extending to medium sand up to 7 m away during the monsoon. A log-linear decline in E. coli and Bacteroidales concentrations with distance along the transects in the early monsoon indicates that ponds excavated in medium sand were the likely source of contamination. Spatial removal rates ranged from 0.5 to 1.3 log(10)/m. After the ponds were artificially filled with groundwater to simulate the impact of a rain storm, E. coli levels increased near a pond recently excavated in medium sand, but no others. These observations show that adjacent sediment grain-size and how recently a pond was excavated influence the how much fecal contamination ponds receiving latrine effluents contribute to neighboring groundwater.

  7. A Molecular MST Approach to Investigate Fecal Indicator Bacteria in Bioaerosols, Bathing Water, Seaweed Wrack, and Sand at Recreational Beaches

    NASA Astrophysics Data System (ADS)

    Thoren, K. M.; Sinigalliano, C. D.

    2016-02-01

    Despite numerous cases of beach bacteria affecting millions of people worldwide, the persistence of the bacteria populations in coastal areas is still not well understood. The purpose of this study was to test the levels of persistence of Fecal Indicating Bacteria (FIB) of enterococci, Escherichia coli, and Human-source Bacteroidales, within the intertidal "swash zone" and the deeper waist zone in which people commonly bathe and play. In addition, the study sought to determine if these bacterial contaminants may also be found in aerosols at the beach. Measuring solar insolation in relation to bacterial persistence in seaweed wrack was used to determine if sunlight plays a role in modifying concentrations of FIB at the beach. Light intensity measured by a solar photometer and air quality measured by aerosol plate counts and qPCR Microbial Source Tracking (MST) was compared to varying locations where the beach samples were collected. Results from water samples demonstrate that bacteria measured using plate counts and qPCR were indeed higher within the swash zone than in the waist zone. This is in contrast with the way that the EPA currently measures and determines the public safety of beach waters. They commonly measure the waist zone, but disregard the swash zone. Results from beach bio-aerosol samples showed a wide variety of fungi and bacteria in the beach air, and qPCR MST analysis of these bio-aerosols showed the presence of FIBs such as enterococci on several of the aerosol collection plates. This emphasizes the need to collect samples from the entire beach instead of just measuring at an isolated area, and that exposure to microbial contaminants may include bathing water, beach sand, seaweed wrack, and bio-aerosols. Thus, the data reveals a potential way to identify harmful levels of bacteria and dangerous levels of poor air quality at recreational beaches. These results expound the need for broader assessment of potential beach contamination, not only the

  8. Spatial and temporal variability of fecal indicator bacteria in an urban stream under different meteorological regimes.

    PubMed

    Cha, Sung Min; Lee, Seung Won; Park, Yong Eun; Cho, Kyung Hwa; Lee, Seungyoon; Kim, Joon Ha

    2010-01-01

    As a representative urban stream in Korea, the Gwangju (GJ) stream suffers from chronic fecal contamination. In this study, to characterize levels of fecal pollution in the GJ stream, the monthly monitoring data for seven years (from 2001 to 2007) and the hourly monitoring data from two field experiments were examined with respect to seasonal/daily variations and spatial distribution under wet and dry weather conditions. This research revealed that concentrations of fecal indicator bacteria strongly varied depending on the prevalent meteorological conditions. That is, during the dry daytime, fecal indicator bacteria concentrations decreased due to inactivation from solar irradiation, but rapidly increased in the absence of sunlight, suggesting external source inputs. In addition, bacterial concentrations substantially increased during rainfall events, due probably to a major contribution from combined sewer overflow. The observations in this study can be useful for implementing fecal pollution management strategies and for predicting fecal contamination as a function of meteorological conditions.

  9. Concentrations of fecal coliform bacteria in creeks, Anchorage, Alaska, August and September 1998

    USGS Publications Warehouse

    Dorava, Joseph M.; Love, Andra

    1999-01-01

    Water samples were collected from five creeks in undeveloped, semi-developed, and developed areas of Anchorage, Alaska, during August and September 1998 to determine concentrations of fecal coliform bacteria. In undeveloped areas of Ship, Chester, and Campbell Creeks, and the semi-developed area of Rabbit Creek, concentrations of fecal coliform bacteria ranged from less than 1 to 16 colonies per 100 milliliters of water. In the semi-developed area of Little Rabbit Creek, concentrations ranged from 30 to 860 colonies per 100 milliliters of water. In developed areas of the creeks, concentrations of fecal coliform bacteria ranged from 6 to 80 colonies per 100 milliliters of water.

  10. Fate and Enumeration Problems of Fecal Coliform Bacteria in Runoff Waters from Terrestrial Ecosystems.

    DTIC Science & Technology

    1980-09-01

    manure or wastewater sludges, might exceed present point -source criteria for fecal coliform bacteria. i56 b. Determining the bacterial sorptive...fecal wastes: application rttes were varied to determine best treatment conditions. The fecal colifo’.. (N-PC) test isolates were identified to... determine their origin, fate, and evume, ation problem. Modifications to (Co tin OD 3 Wsee or Iy Rev asisesma Unclassified 11YNT CLMSSPICATIbS. OF YW" P0"t

  11. Physical dynamics controlling variability in nearshore fecal pollution: fecal indicator bacteria as passive particles.

    PubMed

    Rippy, M A; Franks, P J S; Feddersen, F; Guza, R T; Moore, D F

    2013-01-15

    We present results from a 5-h field program (HB06) that took place at California's Huntington State Beach. We assessed the importance of physical dynamics in controlling fecal indicator bacteria (FIB) concentrations during HB06 using an individual based model including alongshore advection and cross-shore variable horizontal diffusion. The model was parameterized with physical (waves and currents) and bacterial (Escherichia coli and Enterococcus) observations made during HB06. The model captured surfzone FIB dynamics well (average surfzone model skill: 0.84 {E. coli} and 0.52 {Enterococcus}), but fell short of capturing offshore FIB dynamics. Our analyses support the hypothesis that surfzone FIB variability during HB06 was a consequence of southward advection and diffusion of a patch of FIB originating north of the study area. Offshore FIB may have originated from a different, southern, source. Mortality may account for some of the offshore variability not explained by the physical model. Copyright © 2012 Elsevier Ltd. All rights reserved.

  12. Distribution of Fecal Indicator Bacteria along the Malibu, California, Coastline

    USGS Publications Warehouse

    Izbicki, John

    2011-01-01

    Each year, over 550 million people visit California's public beaches. To protect beach-goers from exposure to waterborne disease, California state law requires water-quality monitoring for fecal indicator bacteria (FIB), such as enterococci and Escherichia coli (E. coli), at beaches having more than 50,000 yearly visitors. FIB are used to assess the microbiological quality of water because, although not typically disease causing, they are correlated with the occurrence of certain waterborne diseases. Tests show that FIB concentrations occasionally exceed U.S. Environmental Protection Agency (USEPA) public health standards for recreational water in Malibu Lagoon and at several Malibu beaches (Regional Water Quality Control Board, 2009). Scientists from the U.S. Geological Survey's (USGS) California Water Science Center are doing a study to identify the distribution and sources of FIB in coastal Malibu waters (fig. 1). The study methods were similar to those used in a study of FIB contamination on beaches in the Santa Barbara, California, area (Izbicki and others, 2009). This report describes the study approach and presents preliminary results used to evaluate the distribution and source of FIB in the Malibu area. Results of this study will help decision-makers address human health issues associated with FIB contamination in Malibu, and the methods used in this study can be used in other coastal areas affected by FIB contamination.

  13. Predicting Fecal Indicator Bacteria Concentrations in the South Fork Broad River Watershed Using Virtual Beach

    EPA Science Inventory

    Virtual Beach (VB) is a decision support tool that constructs site-specific statistical models to predict fecal indicator bacteria (FIB) at recreational beaches. Although primarily designed for making decisions regarding beach closures or issuance of swimming advisories based on...

  14. Predicting Fecal Indicator Bacteria Concentrations in the South Fork Broad River Watershed Using Virtual Beach

    EPA Science Inventory

    Virtual Beach (VB) is a decision support tool that constructs site-specific statistical models to predict fecal indicator bacteria (FIB) at recreational beaches. Although primarily designed for making decisions regarding beach closures or issuance of swimming advisories based on...

  15. Predicting Fecal Indicator Bacteria Fate and Removal in Urban Stormwater at the Watershed Scale

    NASA Astrophysics Data System (ADS)

    Wolfand, J.; Hogue, T. S.; Luthy, R. G.

    2016-12-01

    Urban stormwater is a major cause of water quality impairment, resulting in surface waters that fail to meet water quality standards and support their designated uses. Of the many stormwater pollutants, fecal indicator bacteria are particularly important to track because they are directly linked to pathogens which jeopardize public health; yet, their fate and transport in urban stormwater is poorly understood. Monitoring fecal bacteria in stormwater is possible, but due to the high variability of fecal indicators both spatially and temporally, single grab or composite samples do not fully capture fecal indicator loading. Models have been developed to predict fecal indicator bacteria at the watershed scale, but they are often limited to agricultural areas, or areas that receive frequent rainfall. Further, it is unclear whether best management practices (BMPs), such as bioretention or engineered wetlands, are able to reduce bacteria to meet water quality standards at watershed outlets. This research seeks to develop a model to predict fecal indicator bacteria in urban stormwater in a semi-arid climate at the watershed scale. Using the highly developed Ballona Creek watershed (89 mi2) located in Los Angeles County as a case study, several existing mechanistic models are coupled with a hydrologic model to predict fecal indicator concentrations (E. coli, enterococci, fecal coliform, and total coliform) at the outfall of Ballona Creek watershed, Santa Monica Bay. The hydrologic model was developed using InfoSWMM Sustain, calibrated for flow from WY 1998-2006 (NSE = 0.94; R2 = 0.95), and validated from WY 2007-2015 (NSE = 0.93; R2 = 0.95). The developed coupled model is being used to predict fecal indicator fate and transport and evaluate how BMPs can be optimized to reduce fecal indicator loading to surface waters and recreational beaches.

  16. Isolation of Fecal Coliform Bacteria from the Diamondback Terrapin (Malaclemys terrapin centrata)

    PubMed Central

    Harwood, Valerie J.; Butler, Joseph; Parrish, Danny; Wagner, Victoria

    1999-01-01

    Total and fecal coliform bacteria were isolated from the cloaca and feces of the estuarine diamondback terrapin. The majority of samples contained fecal coliforms. Escherichia coli was the predominant fecal coliform species isolated, and members of the genus Salmonella were isolated from 2 of 39 terrapins. Fecal coliform numbers are used to regulate shellfish harvests, and diamondback terrapins inhabit the brackish-water habitats where oyster beds are found; therefore, these findings have implications for the efficacy of current regulatory parameters in shellfishing waters. PMID:9925633

  17. Isolation of fecal coliform bacteria from the diamondback terrapin (Malaclemys terrapin centrata).

    PubMed

    Harwood, V J; Butler, J; Parrish, D; Wagner, V

    1999-02-01

    Total and fecal coliform bacteria were isolated from the cloaca and feces of the estuarine diamondback terrapin. The majority of samples contained fecal coliforms. Escherichia coli was the predominant fecal coliform species isolated, and members of the genus Salmonella were isolated from 2 of 39 terrapins. Fecal coliform numbers are used to regulate shellfish harvests, and diamondback terrapins inhabit the brackish-water habitats where oyster beds are found; therefore, these findings have implications for the efficacy of current regulatory parameters in shellfishing waters.

  18. Source specific fecal bacteria modeling using soil and water assessment tool model.

    PubMed

    Parajuli, Prem B; Mankin, Kyle R; Barnes, Philip L

    2009-01-01

    Fecal bacteria can contaminate water and result in illness or death. It is often difficult to accurately determine sources of fecal bacteria contamination, but bacteria source tracking can help identify non-point sources of fecal bacteria such as livestock, humans and wildlife. The Soil and Water Assessment Tool (SWAT) microbial sub-model 2005 was used to evaluate source-specific fecal bacteria using three years (2004-2006) of observed modified deterministic probability of bacteria source tracking data, as well as measure hydrologic and water quality data. This study modeled source-specific bacteria using a model previously calibrated for flow, sediment and total fecal coliform bacteria (FCB) concentration. The SWAT model was calibrated at the Rock Creek sub-watershed, validated at the Deer Creek sub-watershed, and verified at the Auburn sub-watershed and then at the entire Upper Wakarusa watershed for predicting daily flow, sediment, nutrients, total fecal bacteria, and source-specific fecal bacteria. Watershed characteristics for livestock, humans, and wildlife fecal bacterial sources were first modeled together then with three separate sources and combinations of source-specific FCB concentration: livestock and human, livestock and wildlife and human and wildlife. Model results indicated both coefficient of determination (R(2)) and Nash-Sutcliffe Efficiency Index (E) parameters ranging from 0.52 to 0.84 for daily flow and 0.50-0.87 for sediment (good to very good agreement); 0.14-0.85 for total phosphorus (poor to very good agreement); -3.55 to 0.79 for total nitrogen (unsatisfactory to very good agreement) and -2.2 to 0.52 for total fecal bacteria (unsatisfactory to good agreement). Model results generally determined decreased agreement for each single source of bacteria (R(2) and E range from -5.03 to 0.39), potentially due to bacteria source tracking (BST) uncertainty and spatial variability. This study contributes to new knowledge in bacteria modeling and

  19. Seasonal variation of fecal indicator bacteria in storm events within the US stormwater database.

    PubMed

    Pan, Xubin; Jones, Kim D

    2012-01-01

    Bacteria are one of the major causes of surface water impairments in the USA. Over the past several years, best management practices, including detention basins, manufactured devices, grass swales, filters and bioretention cells have been used to remove bacteria and other pollutants from stormwater runoff. However, there are data gaps in the comprehensive studies of bacteria concentrations in stormwater runoff. In this paper, the event mean concentration (EMC) of fecal indicator bacteria (Enterococcus, Escherichia coli, fecal Streptococcus group bacteria, and fecal coliform) across the USA was retrieved from the international stormwater best management practices database to analyze the seasonal variations of inflow and outflow event mean concentrations and removal efficiencies. The Kruskal-Wallis test was employed to determine the seasonal variations of bacteria indicator concentrations and removals, and the two-sample Kolmogorov-Smirnov test was used for comparing different seasonal outcomes. The results indicate that all the inflow EMC of FIB in stormwater runoff is above the water quality criteria. The seasonal differences of fecal Streptococcus group bacteria and fecal coliform are significant. Summer has the potential to increase the bacteria EMC and illustrate the seasonal differences.

  20. COMPARATIVE DIVERSITY OF FECAL BACTERIA IN AGRICULTURALLY SIGNIFICANT ANIMALS TO IDENTIFY ALTERNATIVE TARGETS FOR MICROBIAL SOURCE TRACKING

    EPA Science Inventory

    Animals of agricultural significance contribute a large percentage of fecal pollution to waterways via runoff contamination. The premise of microbial source tracking is to utilize fecal bacteria to identify target populations which are directly correlated to specific animal feces...

  1. COMPARATIVE DIVERSITY OF FECAL BACTERIA IN AGRICULTURALLY SIGNIFICANT ANIMALS TO IDENTIFY ALTERNATIVE TARGETS FOR MICROBIAL SOURCE TRACKING

    EPA Science Inventory

    Animals of agricultural significance contribute a large percentage of fecal pollution to waterways via runoff contamination. The premise of microbial source tracking is to utilize fecal bacteria to identify target populations which are directly correlated to specific animal feces...

  2. Sediment and Fecal Indicator Bacteria Loading in a Mixed Land Use Watershed: Contributions from Suspended and Bed Load Transport

    EPA Science Inventory

    Water quality studies that quantify sediment and fecal bacteria loading commonly focus on suspended contaminants transported during high flows. Fecal contaminants in bed sediments are typically ignored and need to be considered because of their potential to increase pathogen load...

  3. Sediment and Fecal Indicator Bacteria Loading in a Mixed Land Use Watershed: Contributions from Suspended and Bed Load Transport

    EPA Science Inventory

    Water quality studies that quantify sediment and fecal bacteria loading commonly focus on suspended contaminants transported during high flows. Fecal contaminants in bed sediments are typically ignored and need to be considered because of their potential to increase pathogen load...

  4. Performance evaluation of canine-associated Bacteroidales assays in a multi-laboratory comparison study

    EPA Science Inventory

    The contribution of fecal pollution from dogs in urbanized areas can be significant and is an often underestimated problem. Microbial source tracking methods (MST) utilizing quantitative PCR of dog-associated gene sequences encoding 16S rRNA of Bacteroidales are a useful tool to ...

  5. Performance evaluation of canine-associated Bacteroidales assays in a multi-laboratory comparison study

    EPA Science Inventory

    The contribution of fecal pollution from dogs in urbanized areas can be significant and is an often underestimated problem. Microbial source tracking methods (MST) utilizing quantitative PCR of dog-associated gene sequences encoding 16S rRNA of Bacteroidales are a useful tool to ...

  6. Occurrence of fecal coliform bacteria in selected streams in Wyoming, 1990-99

    USGS Publications Warehouse

    Clark, Melanie L.; Norris, Jodi R.

    2000-01-01

    The U.S. Geological Survey (USGS), in cooperation with the Wyoming Department of Environmental Quality (WDEQ), is collecting water samples for analysis of fecal coliform bacteria at 18 stream sites as part of a statewide network. Contamination by bacteria of fecal origin in streams where contact recreation is a designated water use is a concern because of potential public-health risk from the presence of enteric pathogens. Fecal coliform concentrations are temporally and spatially variable in Wyoming streams-concentrations ranged from less than 1 to 45,000 colonies per 100 milliliters of water during 1990-99. Fecal coliform concentrations were less than the water-quality criterion of 400 colonies per 100 milliliters in 83 percent of the samples, indicating fecal coliform contamination is not a widespread problem in these Wyoming streams. However, 14 of the 18 monitoring sites had at least one sample in which the fecal coliform concentration exceeded 400 colonies per 100 milliliters at some time during the 10-year period. Fecal coliform concentrations generally are higher during April through September than during October through March. The higher concentrations coincide with the time period when the public-health risk is higher because summer months are when contact recreation use is more likely occurring. Fecal coliform concentrations were positively correlated with discharge and stream temperature and generally were negatively correlated with pH, specific conductance, and dissolved oxygen.

  7. Fecal-indicator bacteria in streams alonga gradient of residential development

    USGS Publications Warehouse

    Frenzel, Steven A.; Couvillion, Charles S.

    2002-01-01

    Fecal-indicator bacteria were sampled at 14 stream sites in Anchorage, Alaska, USA, as part of a study to determine the effects of urbanization on water quality. Population density in the subbasins sampled ranged from zero to 1,750 persons per square kilometer. Higher concentrations of fecal-coliform, E. coli, and enterococci bacteria were measured at the most urbanized sites. Although fecal-indicator bacteria concentrations were higher in summer than in winter, seasonal differences in bacteria concentrations generally were not significant. Areas served by sewer systems had significantly higher fecal-indicator bacteria concentrations than did areas served by septic systems. The areas served by sewer systems also had storm drains that discharged directly to the streams, whereas storm sewers were not present in the areas served by septic systems. Fecal-indicator bacteria concentrations were highly variable over a two-day period of stable streamflow, which may have implications for testing of compliance to water-quality standards.

  8. Micro-zooplankton grazing as a means of fecal bacteria removal in stormwater BMPs.

    PubMed

    Burtchett, Jade M; Mallin, Michael A; Cahoon, Lawrence B

    2017-06-01

    A priority for environmental managers is control of stormwater runoff pollution, especially fecal microbial pollution. This research was designed to determine if fecal bacterial grazing by micro-zooplankton is a significant control on fecal bacteria in aquatic best management practices (BMPs); if grazing differs between a wet detention pond and a constructed wetland; and if environmental factors enhance grazing. Both 3-day grazing tests and 24-h dilution assays were used to determine grazing differences between the two types of BMP. Micro-zooplankton grazing was a stronger bacteria removal mechanism in stormwater wetlands rich in aquatic vegetation compared to a standard wet detention pond, although grazing was important in detention ponds as well. Our experiments indicated that the majority of grazers that fed on fecal bacteria were <20 μm in size. Grazing rates were positively correlated with fecal coliform abundance and increased water temperatures. Enumeration of grazers demonstrated that protozoans were significantly more abundant among wetland vegetation than in open water, and open wetland waters contained more flagellates and dinoflagellates than open wet detention pond waters. Grazing on fecal bacteria in BMPs is enhanced by aquatic vegetation, and grazing in aquatic BMPs in warmer climates should be greater than in cooler climates.

  9. Patterns and sources of fecal coliform bacteria in three streams in Virginia, 1999-2000

    USGS Publications Warehouse

    Hyer, Kenneth; Moyer, Douglas

    2003-01-01

    Surface-water impairment by fecal coliform bacteria is a water-quality issue of national scope and importance. In Virginia, more than 175 stream segments are on the Commonwealth's 1998 303(d) list of impaired waters because of elevated concentrations of fecal coliform bacteria. These fecal coliform-impaired stream segments require the development of total maximum daily load (TMDL) and associated implementation plans, but accurate information on the sources contributing these bacteria usually is lacking. The development of defendable fecal coliform TMDLs and management plans can benefit from reliable information on the bacteria sources that are responsible for the impairment. Bacterial source tracking (BST) recently has emerged as a powerful tool for identifying the sources of fecal coliform bacteria that impair surface waters. In a demonstration of BST technology, three watersheds on Virginia's 1998 303(d) list with diverse land-use practices (and potentially diverse bacteria sources) were studied. Accotink Creek is dominated by urban land uses, Christians Creek by agricultural land uses, and Blacks Run is affected by both urban and agricultural land uses. During the 20-month field study (March 1999?October 2000), water samples were collected from each stream during a range of flow conditions and seasons. For each sample, specific conductance, dissolved oxygen concentration, pH, turbidity, flow, and water temperature were measured. Fecal coliform concentrations of each water sample were determined using the membrane filtration technique. Next, Escherichia coli (E. coli) were isolated from the fecal coliform bacteria and their sources were identified using ribotyping (a method of 'genetic fingerprinting'). Study results provide enhanced understanding of the concentrations and sources of fecal coliform bacteria in these three watersheds. Continuum sampling (sampling along the length of the streams) indicated that elevated concentrations of fecal coliform bacteria

  10. Inactivation of fecal bacteria in drinking water by solar heating.

    PubMed Central

    Joyce, T M; McGuigan, K G; Elmore-Meegan, M; Conroy, R M

    1996-01-01

    We report simulations of the thermal effect of strong equatorial sunshine on water samples contaminated with high populations of fecal coliforms. Water samples, heavily contaminated with a wild-type strain of Escherichia coli (starting population = 20 x 10(5) CFU/ml), are heated to those temperatures recorded for 2-liter samples stored in transparent plastic bottles and exposed to full Kenyan sunshine (maximum water temperature, 55 degrees C). The samples are completely disinfected within 7 h, and no viable E. coli organisms are detected at either the end of the experiment or a further 12 h later, showing that no bacterial recovery has occurred. The feasibility of employing solar disinfection for highly turbid, fecally contaminated water is discussed. PMID:8593045

  11. Inactivation of fecal bacteria in drinking water by solar heating.

    PubMed

    Joyce, T M; McGuigan, K G; Elmore-Meegan, M; Conroy, R M

    1996-02-01

    We report simulations of the thermal effect of strong equatorial sunshine on water samples contaminated with high populations of fecal coliforms. Water samples, heavily contaminated with a wild-type strain of Escherichia coli (starting population = 20 x 10(5) CFU/ml), are heated to those temperatures recorded for 2-liter samples stored in transparent plastic bottles and exposed to full Kenyan sunshine (maximum water temperature, 55 degrees C). The samples are completely disinfected within 7 h, and no viable E. coli organisms are detected at either the end of the experiment or a further 12 h later, showing that no bacterial recovery has occurred. The feasibility of employing solar disinfection for highly turbid, fecally contaminated water is discussed.

  12. Traditional and Molecular Analyses for Fecal Indicator Bacteria in Non-point Source Subtropical Recreational Marine Waters

    PubMed Central

    Sinigalliano, Christopher D.; Fleisher, Jay M.; Gidley, Maribeth L.; Solo-Gabriele, Helena M.; Shibata, Tomoyuki; Plano, Lisa R.W.; Elmir, Samir M.; Wanless, David; Bartkowiak, Jakub; Boiteau, Rene; Withum, Kelly; Abdelzaher, Amir M.; He, Guoqing; Ortega, Cristina; Zhu, Xiaofang; Wright, Mary E.; Kish, Jonathan; Hollenbeck, Julie; Scott, Troy; Backer, Lorraine C.; Fleming, Lora E.

    2010-01-01

    The use of enterococci as the primary fecal indicator bacteria (FIB) for the determination of recreational water safety has been questioned, particularly in sub/tropical marine waters without known point sources of sewage. Alternative FIB (such as the Bacteroidales group) and alternative measurement methods (such as rapid molecular testing) have been proposed to supplement or replace current marine water quality testing methods which require culturing enterococci. Moreover, environmental parameters have also been proposed to supplement current monitoring programs. The objective of this study was to evaluate the health risks to humans from exposure to subtropical recreational marine waters with no known point source. The study reported symptoms between one set of human subjects randomly assigned to marine water exposure with intensive environmental monitoring compared with other subjects who did not have exposure. In addition, illness outcomes among the exposed bathers were compared to levels of traditional and alternative FIB (as measured by culture-based and molecular-based methods), and compared to easily measured environmental parameters. Results demonstrated an increase in self-reported gastrointestinal, respiratory and skin illnesses among bathers vs. non-bathers. Among the bathers, a dose-response relationship by logistic regression modeling was observed for skin illness, where illness was positively related to enterococci enumeration by membrane filtration (odds ratio =1.46 [95% confidence interval=0.97-2.21] per increasing log10 unit of enterococci exposure) and positively related to 24 hour antecedent rain fall (1.04 [1.01 – 1.07] per increasing millimeters of rain). Acute febrile respiratory illness was inversely related to water temperature (0.74 [0.56-0.98] per increasing degree of water temperature). There were no significant dose response relationships between report of human illness and any of the other FIB or environmental measures. Therefore, for

  13. Impact of Population and Latrines on Fecal Contamination of Ponds in Rural Bangladesh

    PubMed Central

    Knappett, Peter S. K.; Escamilla, Veronica; Layton, Alice; McKay, Larry D.; Emch, Michael; Williams, Daniel E.; Huq, Md. R.; Alam, Md. J.; Farhana, Labony; Mailloux, Brian J.; Ferguson, Andy; Sayler, Gary S.; Ahmed, Kazi M.; van Geen, Alexander

    2011-01-01

    A majority of households in Bangladesh rely on pond water for hygiene. Exposure to pond water fecal contamination could therefore still contribute to diarrheal disease despite the installation of numerous tubewells for drinking. The objectives of this study are to determine the predominant sources (human or livestock) of fecal pollution in ponds and examine the association between local population, latrine density, latrine quality and concentrations of fecal bacteria and pathogens in pond water. Forty-three ponds were analyzed for E. coli using culture-based methods and E. coli, Bacteroidales and adenovirus using quantitative PCR. Population and sanitation spatial data were collected and measured against pond fecal contamination. Humans were the dominant source of fecal contamination in 79% of the ponds according to Bacteroidales measurements. Ponds directly receiving latrine effluent had the highest concentrations of fecal indicator bacteria (up to 106 Most Probable Number (MPN) of culturable E. coli per 100 mL). Concentrations of fecal indicator bacteria correlated with population surveyed within a distance of 30-70 m (p<0.05) and total latrines surveyed within 50-70 m (p<0.05). Unsanitary latrines (visible effluent or open pits) within the pond drainage basin were also significantly correlated to fecal indicator concentrations (p<0.05). Water in the vast majority of the surveyed ponds contained unsafe levels of fecal contamination attributable primarily to unsanitary latrines, and to lesser extent to sanitary latrines and cattle. Since the majority of fecal pollution is derived from human waste, continued use of pond water could help explain the persistence of diarrheal disease in rural South Asia. PMID:21632095

  14. Quantitative role of shrimp fecal bacteria in organic matter fluxes in a recirculating shrimp aquaculture system.

    PubMed

    Beardsley, Christine; Moss, Shaun; Malfatti, Francesca; Azam, Farooq

    2011-07-01

    Microorganisms play integral roles in the cycling of carbon (C) and nitrogen (N) in recirculating aquaculture systems (RAS) for fish and shellfish production. We quantified the pathways of shrimp fecal bacterial activities and their role in C- and N-flux partitioning relevant to culturing Pacific white shrimp, Penaeus (Litopenaeus) vannamei, in RAS. Freshly produced feces from P. vannamei contained 0.6-7 × 10(10) bacteria g(-1) dry wt belonging to Bacteroidetes (7%), Alphaproteobacteria (4%), and, within the Gammaproteobacteria, almost exclusively to the genus Vibrio (61%). Because of partial disintegration of the feces (up to 27% within 12 h), the experimental seawater became inoculated with fecal bacteria. Bacteria grew rapidly in the feces and in the seawater, and exhibited high levels of aminopeptidase, chitinase, chitobiase, alkaline phosphatase, α- and β-glucosidase, and lipase activities. Moreover, fecal bacteria enriched the protein content of the feces within 12 h, potentially enriching the feces for the coprophagous shrimp. The bacterial turnover time was much faster in feces (1-10 h) than in mature RAS water (350 h). Thus, shrimp fecal bacteria not only inoculate RAS water but also contribute to bacterial abundance and productivity, and regulate system processes important for shrimp health.

  15. Use of barcoded pyrosequencing and shared OTUs to determine sources of fecal bacteria in watersheds.

    PubMed

    Unno, Tatsuya; Jang, Jeonghwan; Han, Dukki; Kim, Joon Ha; Sadowsky, Michael J; Kim, Ok-Sun; Chun, Jongsik; Hur, Hor-Gil

    2010-10-15

    While many current microbial source tracking (MST) methods rely on the use of specific molecular marker genes to identify sources of fecal contamination, these methods often fail to determine all point and nonpoint contributors of fecal inputs into waterways. In this study, we developed a new library-dependent MST method that uses pyrosequencing-derived shared operational taxonomy units (OTUs) to define sources of fecal contamination in waterways. A total 56,841 pyrosequencing reads of 16S rDNA obtained from the feces of humans and animals were evaluated and used to compare fecal microbial diversity in three freshwater samples obtained from the Yeongsan river basin in Jeonnam Province, South Korea. Sites included an urbanized agricultural area (Y1) (Escherichia coli counts ≥ 1600 CFU/100 mL), an open area (Y2) with no major industrial activities (940 CFU/100 mL), and a typical agricultural area (Y3) (≥ 1600 CFU/100 mL). Data analyses indicated that the majority of bacteria in the feces of humans and domesticated animals were comprised of members of the phyla Bacteroidetes or Firmicutes, whereas the majority of bacteria in wild goose feces and freshwater samples were classified to the phylum Proteobacteria. Analysis of OTUs shared between the fecal and environmental samples suggested that the potential sources of the fecal contamination at the sites were of human and swine origin. Quantification of fecal contamination was also examined by comparing the density of pyrosequencing reads in each fecal sample within shared OTUs. Taken together, our results indicated that analysis of shared OTUs derived from barcoded pyrosequencing reads provide the necessary resolution and discrimination to be useful as a next generation platform for microbial source tracking studies.

  16. Natural soil reservoirs for human pathogenic and fecal indicator bacteria

    USGS Publications Warehouse

    Boschiroli, Maria L; Falkinham, Joseph; Favre-Bonte, Sabine; Nazaret, Sylvie; Piveteau, Pascal; Sadowsky, Michael J.; Byappanahalli, Muruleedhara; Delaquis, Pascal; Hartmann, Alain

    2016-01-01

    Soils receive inputs of human pathogenic and indicator bacteria through land application of animal manures or sewage sludge, and inputs by wildlife. Soil is an extremely heterogeneous substrate and contains meso- and macrofauna that may be reservoirs for bacteria of human health concern. The ability to detect and quantify bacteria of human health concern is important in risk assessments and in evaluating the efficacy of agricultural soil management practices that are protective of crop quality and protective of adjacent water resources. The present chapter describes the distribution of selected Gram-positive and Gram-negative bacteria in soils. Methods for detecting and quantifying soilborne bacteria including extraction, enrichment using immunomagnetic capture, culturing, molecular detection and deep sequencing of metagenomic DNA to detect pathogens are overviewed. Methods for strain phenotypic and genotypic characterization are presented, as well as how comparison with clinical isolates can inform the potential for human health risk.

  17. Detection of fecal bacteria and source tracking identifiers in environmental waters using rRNA-based RT-qPCR and rDNA-based qPCR assays.

    PubMed

    Pitkänen, Tarja; Ryu, Hodon; Elk, Michael; Hokajärvi, Anna-Maria; Siponen, Sallamaari; Vepsäläinen, Asko; Räsänen, Pia; Santo Domingo, Jorge W

    2013-01-01

    In this study, we evaluated the use of RT-qPCR assays targeting rRNA gene sequences for the detection of fecal bacteria in water samples. We challenged the RT-qPCR assays against RNA extracted from sewage effluent (n = 14), surface water (n = 30), and treated source water (n = 15) samples. Additionally, we applied the same assays using DNA as the qPCR template. The targeted fecal bacteria were present in most of the samples tested, although in several cases, the detection frequency increased when RNA was used as the template. For example, the majority of samples that tested positive for E. coli and Campylobacter spp. in surface waters, and for human-specific Bacteroidales, E. coli, and Enterococcus spp. in treated source waters were only detected when rRNA was used as the original template. The difference in detection frequency using rRNA or rDNA (rRNA gene) was sample- and assay-dependent, suggesting that the abundance of active and nonactive populations differed between samples. Statistical analyses for each population exhibiting multiple quantifiable results showed that the rRNA copy numbers were significantly higher than the rDNA counterparts (p < 0.05). Moreover, the detection frequency of rRNA-based assays were in better agreement with the culture-based results of E. coli, intestinal enterococci, and thermotolerant Campylobacter spp. in surface waters than that of rDNA-based assays, suggesting that rRNA signals were associated to active bacterial populations. Our data show that using rRNA-based approaches significantly increases detection sensitivity for common fecal bacteria in environmental waters. These findings have important implications for microbial water quality monitoring and public health risk assessments.

  18. INTERNAL AMPLIFICATION CONTROL FOR USE IN QUANTITATIVE POLYMERASE CHAIN REACTION FECAL INDICATOR BACTERIA ASSAYS

    EPA Science Inventory

    Quantitative polymerase chain reaction (QPCR) can be used as a rapid method for detecting fecal indicator bacteria. Because false negative results can be caused by PCR inhibitors that co-extract with the DNA samples, an internal amplification control (IAC) should be run with eac...

  19. Interlaboratory Comparison of Real-time PCR Protocols for Quantification of General Fecal Indicator Bacteria

    EPA Science Inventory

    The application of quantitative real-time PCR (qPCR) technologies for the rapid identification of fecal bacteria in environmental waters is being considered for use as a national water quality metric in the United States. The transition from research tool to a standardized proto...

  20. INTERNAL AMPLIFICATION CONTROL FOR USE IN QUANTITATIVE POLYMERASE CHAIN REACTION FECAL INDICATOR BACTERIA ASSAYS

    EPA Science Inventory

    Quantitative polymerase chain reaction (QPCR) can be used as a rapid method for detecting fecal indicator bacteria. Because false negative results can be caused by PCR inhibitors that co-extract with the DNA samples, an internal amplification control (IAC) should be run with eac...

  1. Hydrological modeling of fecal indicator bacteria in a tropical mountain catchment

    USDA-ARS?s Scientific Manuscript database

    The occurrence of pathogen bacteria in surface waters is a threat to public health worldwide. In particular, inadequate sanitation resulting in high contamination of surface water with pathogens of fecal origin is a serious issue in developing countries such as Lao P.D.R. Despite the health implicat...

  2. Temporal Synchronization Analysis for Improving Regression Modeling of Fecal Indicator Bacteria Levels

    EPA Science Inventory

    Multiple linear regression models are often used to predict levels of fecal indicator bacteria (FIB) in recreational swimming waters based on independent variables (IVs) such as meteorologic, hydrodynamic, and water-quality measures. The IVs used for these analyses are traditiona...

  3. DNA analysis of fecal bacteria to augment an epikarst dye trace study at Crump's Cave, Kentucky

    USDA-ARS?s Scientific Manuscript database

    A rainfall simulation experiment was performed to investigate the transport behavior of fecal-derived bacteria through shallow karst soils and through the epikarst. The experiment was conducted at Cave Springs Cavern located just south of Mammoth Cave National Park on the Sinkhole Plain of South Cen...

  4. Temporal Synchronization Analysis for Improving Regression Modeling of Fecal Indicator Bacteria Levels

    EPA Science Inventory

    Multiple linear regression models are often used to predict levels of fecal indicator bacteria (FIB) in recreational swimming waters based on independent variables (IVs) such as meteorologic, hydrodynamic, and water-quality measures. The IVs used for these analyses are traditiona...

  5. Interlaboratory Comparison of Real-time PCR Protocols for Quantification of General Fecal Indicator Bacteria

    EPA Science Inventory

    The application of quantitative real-time PCR (qPCR) technologies for the rapid identification of fecal bacteria in environmental waters is being considered for use as a national water quality metric in the United States. The transition from research tool to a standardized proto...

  6. Decay of Fecal Indicator Bacteria and Microbial Source Tracking Markers in Cattle Feces

    EPA Science Inventory

    The survival of fecal indicator bacteria (FIB) and microbial source tracking (MST) markers in water microcosms and manure amended soils has been well documented; however, little is known about the survival of MST markers in bovine feces deposited on pastures. We conducted a study...

  7. Decay of Fecal Indicator Bacteria and Microbial Source Tracking Markers in Cattle Feces

    EPA Science Inventory

    The survival of fecal indicator bacteria (FIB) and microbial source tracking (MST) markers in water microcosms and manure amended soils has been well documented; however, little is known about the survival of MST markers in bovine feces deposited on pastures. We conducted a study...

  8. Comparison of FecalSwab and ESwab Devices for Storage and Transportation of Diarrheagenic Bacteria

    PubMed Central

    Kaukoranta, Suvi-Sirkku

    2014-01-01

    Using a collection (n = 12) of ATCC and known stock isolates, as well as 328 clinical stool specimens, we evaluated the ESwab and the new FecalSwab liquid-based microbiology (LBM) devices for storing and transporting diarrheagenic bacteria. The stock isolates were stored in these swab devices up to 48 h at refrigeration (4°C) or room (∼25°C) temperature and up to 3 months at −20°C or −70°C. With the clinical stool specimens, the performances of the ESwab and FecalSwab were compared to those of routinely used transport systems (Amies gel swabs and dry containers). At a refrigeration temperature, all isolates survived in FecalSwab up to 48 h, while in ESwab, only 10 isolates (83.3%) out of 12 survived. At −70°C, all isolates in FecalSwab were recovered after 3 months of storage, whereas in ESwab, none of the isolates were recovered. At −20°C, neither of the swab devices preserved the viability of stock isolates after 2 weeks of storage, and at room temperature, 7 (58.3%) of the stock isolates were recovered in both transport devices after 48 h. Of the 328 fecal specimens, 44 (13.4%) were positive for one of the common diarrheagenic bacterial species with all transport systems used. Thus, the suitability of the ESwab and FecalSwab devices for culturing fresh stools was at least equal to those of the Amies gel swabs and dry containers. Although the ESwab was shown to be an option for collecting and transporting fecal specimens, the FecalSwab device had clearly better preserving properties under different storage conditions. PMID:24740083

  9. Comparison of FecalSwab and ESwab devices for storage and transportation of Diarrheagenic bacteria.

    PubMed

    Hirvonen, Jari J; Kaukoranta, Suvi-Sirkku

    2014-07-01

    Using a collection (n = 12) of ATCC and known stock isolates, as well as 328 clinical stool specimens, we evaluated the ESwab and the new FecalSwab liquid-based microbiology (LBM) devices for storing and transporting diarrheagenic bacteria. The stock isolates were stored in these swab devices up to 48 h at refrigeration (4°C) or room (∼25°C) temperature and up to 3 months at -20°C or -70°C. With the clinical stool specimens, the performances of the ESwab and FecalSwab were compared to those of routinely used transport systems (Amies gel swabs and dry containers). At a refrigeration temperature, all isolates survived in FecalSwab up to 48 h, while in ESwab, only 10 isolates (83.3%) out of 12 survived. At -70°C, all isolates in FecalSwab were recovered after 3 months of storage, whereas in ESwab, none of the isolates were recovered. At -20°C, neither of the swab devices preserved the viability of stock isolates after 2 weeks of storage, and at room temperature, 7 (58.3%) of the stock isolates were recovered in both transport devices after 48 h. Of the 328 fecal specimens, 44 (13.4%) were positive for one of the common diarrheagenic bacterial species with all transport systems used. Thus, the suitability of the ESwab and FecalSwab devices for culturing fresh stools was at least equal to those of the Amies gel swabs and dry containers. Although the ESwab was shown to be an option for collecting and transporting fecal specimens, the FecalSwab device had clearly better preserving properties under different storage conditions.

  10. Historical trends and concentrations of fecal coliform bacteria in the Brandywine Creek basin, Chester County, Pennsylvania

    USGS Publications Warehouse

    Town, D.A.

    2001-01-01

    The Brandywine Creek in Chester County is used for recreation and as an important source of drinking water. For this study, 40 sites were established for collection of water samples for analysis of fecal coliform and Escherichia coli bacteria in 1998-99. Samples were collected during base-flow conditions and during five storms in which rainfall exceeded 0.5 inch. During base- flow conditions, the median concentrations of fecal coliform bacteria exceeded 200 col/100 mL at 26 of the 40 sites (65 percent). During stormflow conditions, the median concentration of fecal coliform bacteria exceeded the Pennsylvania Department of Environmental Protection (PaDEP) criterion of 200 col/100 mL at 30 of 33 sites sampled (91 percent). Trends in fecal coliform bacteria concentrations were downward for the period 1973-99 at three long-term water-quality monitor stations, the result of upgrades in wastewater treatment plants, decreases in point-source discharges, and a decrease in agricultural land. A positive relation exists between streamflow and concentrations of fecal coliform bacteria at two of the long-term stations, but concentrations are elevated in base flow and stormflow at all three stations. Factors affecting bacteria concentrations in the Brandywine Creek Basin include nonpoint-source contaminants, reservoirs, seasonality, and stormflow. Nonpoint sources of bacterial contamination in the basin include, but are not limited to, land-surface runoff, urbanization, agricultural processes, groundwater contamination, and wildlife. Bacteria concentrations in streams that flow directly from the reservoirs are much lower than the concentrations in the streams flowing into the reservoirs. During March, April, May, October, and November, the Brandywine Creek tends to have lower water temperatures and bacteria concentrations than during June, July, August, and September. The 10-year median concentrations of bacteria at West Branch Brandywine Creek at Modena and East Branch

  11. Differential Decomposition of Bacterial and Viral Fecal ...

    EPA Pesticide Factsheets

    Understanding the decomposition of microorganisms associated with different human fecal pollution types is necessary for proper implementation of many water qualitymanagement practices, as well as predicting associated public health risks. Here, thedecomposition of select cultivated and molecular indicators of fecal pollution originating from fresh human feces, septage, and primary effluent sewage in a subtropical marine environment was assessed over a six day period with an emphasis on the influence of ambient sunlight and indigenous microbiota. Ambient water mixed with each fecal pollution type was placed in dialysis bags and incubated in situ in a submersible aquatic mesocosm. Genetic and cultivated fecal indicators including fecal indicator bacteria (enterococci, E. coli, and Bacteroidales), coliphage (somatic and F+), Bacteroides fragilis phage (GB-124), and human-associated geneticindicators (HF183/BacR287 and HumM2) were measured in each sample. Simple linearregression assessing treatment trends in each pollution type over time showed significant decay (p ≤ 0.05) in most treatments for feces and sewage (27/28 and 32/40, respectively), compared to septage (6/26). A two-way analysis of variance of log10 reduction values for sewage and feces experiments indicated that treatments differentially impact survival of cultivated bacteria, cultivated phage, and genetic indicators. Findings suggest that sunlight is critical for phage decay, and indigenous microbio

  12. Fecal pathogen pollution: sources and patterns in water and sediment samples from the upper Cook Inlet, Alaska ecosystem.

    PubMed

    Norman, Stephanie A; Hobbs, Roderick C; Wuertz, Stefan; Melli, Ann; Beckett, Laurel A; Chouicha, Nadira; Kundu, Arti; Miller, Woutrina A

    2013-05-01

    Fecal pathogens are transported from a variety of sources in multi-use ecosystems such as upper Cook Inlet (CI), Alaska, which includes the state's urban center and is highly utilized by humans and animals. This study used a novel water quality testing approach to evaluate the presence and host sources of potential fecal pathogens in surface waters and sediments from aquatic ecosystems in upper CI. Matched water and sediment samples, along with effluent from a municipal wastewater treatment facility, were screened for Salmonella spp., Vibrio spp., Cryptosporidium spp., Giardia spp., and noroviruses. Additionally, Bacteroidales spp. for microbial source tracking, and the fecal indicator bacteria Enterococcus spp. as well as fecal coliforms were evaluated. Overall, Giardia and Vibrio were the most frequently detected potential pathogens, followed by Cryptosporidium and norovirus, while Salmonella was not detected. Sample month, matrix type, and recent precipitation were found to be significant environmental factors for protozoa or host-associated Bacteroidales marker detection, whereas location and water temperature were not. The relative contribution of host-associated markers to total fecal marker concentration was estimated using a Monte Carlo method, with the greatest relative contribution to the Bacteroidales marker concentration coming from human sources, while the remainder of the universal fecal host source signal was uncharacterized by available host-associated assays, consistent with wildlife fecal sources. These findings show how fecal indicator and pathogen monitoring, along with identifying contributing host sources, can provide evidence of coastal pathogen pollution and guidance as to whether to target human and/or animal sources for management.

  13. Age-related changes in select fecal bacteria in foals

    USDA-ARS?s Scientific Manuscript database

    Adult horses depend on the microbial community in the hindgut to produce VFAs that are utilized for energy. Microbial colonization in the gastrointestinal tract of foals is essential to develop a healthy symbiotic relationship and prevent proliferation of pathogenic bacteria. However, colonization i...

  14. Association of dry cow therapy with the antimicrobial susceptibility of fecal coliform bacteria in dairy cows.

    PubMed

    Mollenkopf, Dixie F; Glendening, Candace; Wittum, Thomas E; Funk, Julie A; Tragesser, Lesley A; Morley, Paul S

    2010-08-01

    The prophylactic use of intramammary antimicrobial drugs at the end of lactation in dairy cows, known as dry cow therapy (DCT), is widely practiced in US dairy herds. This extremely common use of high-dose, slow-release antimicrobials may influence the ecology of bacterial flora on dairy farms. We investigated the association between the antimicrobial used for intramammary DCT and the relative number of fecal coliform bacteria with reduced susceptibility to three antimicrobial drugs in dairy cattle. Most probable number (MPN) data were estimated from 463 individual fecal samples collected from lactating cows in 15 dairy herds in Ohio, USA. These data were used to calculate the relative number of fecal coliform bacteria with reduced susceptibility to cephalothin, streptomycin, and tetracycline for individual cow samples. The farms included in this project were classified based on DCT, with 8 farms using a cephalosporin-based product and the remaining 7 using a penicillin/streptomycin therapy. Results of a linear mixed model indicate that herds using a cephalosporin DCT had higher (P<0.01) relative numbers of fecal coliform bacteria with reduced susceptibility to cephalothin and streptomycin compared to those using a penicillin/streptomycin intramammary therapy. Relative numbers of fecal coliform bacteria with reduced susceptibility to tetracycline was not associated with DCT. These results suggest that high-dose slow-release antimicrobials applied locally in the udder to populations of dairy cows might influence the antimicrobial susceptibility of the enteric flora. However, the potential animal and public health implications of this result are not clear.

  15. Status and trends of fecal indicator bacteria in two urban watersheds.

    PubMed

    Petersen, Tina M; Suarez, Monica P; Rifai, Hanadi S; Jensen, Paul; Su, Yu-Chun; Stein, Ron

    2006-11-01

    This paper examines bacterial levels and their causes in two Houston bayous (Texas). Buffalo and Whiteoak bayous are two of the most contaminated water bodies in Texas for indicator bacteria, based on the frequency and magnitude of contact recreation water quality exceedances. Examination of historical data indicates frequent exceedances, although some improvement has been made since the 1970s. Statistical analyses showed some correlation between in-stream fecal coliform concentrations and rainfall and with land use. Differences in fecal coliform concentrations were found between high- and low-flow conditions in Whiteoak Bayou, while reservoir releases confounded this relationship in Buffalo Bayou. Wastewater treatment plant effluent was found to make up two-thirds to three-fourths of the median flow in both bayous. Effluent sampling was conducted at 72 of the approximately 140 wastewater treatment plants (WWTPs) in the watersheds, providing evidence that WWTP effluent could act to maintain low-flow concentrations of fecal coliform in the bayous.

  16. Interaction of ambient conditions and fecal coliform bacteria in southern Lake Michigan beach waters: Monitoring program implications

    USGS Publications Warehouse

    Whitman, Richard L.; Nevers, Meredith Becker; Gerovac, Paul J.

    1999-01-01

    Excessive fecal coliform bacteria in public swimming waters can potentially threaten visitor health. Fecal coliform bacteria (1984-1989) and Escherichia coli (1990-1995) density were monitored weekly at the Indiana Dunes National Lakeshore beaches for 12 summers, and park officials closed swimming areas when fecal coliform density exceeded the state water quality criteria (400 CFU fecal coliforms/ 100 ml; 235 CFU E. coli/100 ml water). Due to a 24-hour incubation in the fecal coliform and E. coli assays, beaches were closed the day after collection of high fecal coliform. Our analysis suggests that it is not possible to predict one day's fecal coliform count based on the previous day's results in waters taken from southern Lake Michigan beaches. Dispersal and deposition of bacteria were not uniform among sites or across time apparently due to interactions among environmental variables including rainfall, wind direction, water temperature, and bacteria source. Rainfall combined with northwest winds increased bacteria concentrations. Escherichia coli followed a seasonal trend with similar fluctuations in density among beaches. We suggest that the current beach monitoring protocol is inadequate for predicting fecal coliform density at the time of beach closure, and, subsequently, its use for ensuring visitor safety remains questionable.

  17. Occurrence of fecal-indicator bacteria and protocols for identification of fecal-contamination sources in selected reaches of the West Branch Brandywine Creek, Chester County, Pennsylvania

    USGS Publications Warehouse

    Cinotto, Peter J.

    2005-01-01

    The presence of fecal-indicator bacteria indicates the potential presence of pathogens originating from the fecal matter of warm-blooded animals. These pathogens are responsible for numerous human diseases ranging from common diarrhea to meningitis and polio. The detection of fecal-indicator bacteria and interpretation of the resultant data are, therefore, of great importance to water-resource managers. Current (2005) techniques used to assess fecal contamination within the fluvial environment primarily assess samples collected from the water column, either as grab samples or as depth- and (or) width-integrated samples. However, current research indicates approximately 99 percent of all bacteria within nature exist as attached, or sessile, bacteria. Because of this condition, most current techniques for the detection of fecal contamination, which utilize bacteria, assess only about 1 percent of the total bacteria within the fluvial system and are, therefore, problematic. Evaluation of the environmental factors affecting the occurrence and distribution of bacteria within the fluvial system, as well as the evaluation and modification of alternative approaches that effectively quantify the larger population of sessile bacteria within fluvial sediments, will present water-resource managers with more effective tools to assess, prevent, and (or) eliminate sources of fecal contamination within pristine and impaired watersheds. Two stream reaches on the West Branch Brandywine Creek in the Coatesville, Pa., region were studied between September 2002 and August 2003. The effects of sediment particle size, climatic conditions, aquatic growth, environmental chemistry, impervious surfaces, sediment and soil filtration, and dams on observed bacteria concentrations were evaluated. Alternative approaches were assessed to better detect geographic sources of fecal contamination including the use of turbidity as a surrogate for bacteria, the modification and implementation of sandbag

  18. Selection of Potential Probiotic Lactobacillus with Inhibitory Activity Against Salmonella and Fecal Coliform Bacteria.

    PubMed

    Abhisingha, Mattika; Dumnil, Jureeporn; Pitaksutheepong, Chetsadaporn

    2017-07-15

    Three hundred and sixty presumptive lactic acid bacteria (LAB) isolated from pregnant sows, newborn, suckling, and weaned piglets were preliminarily screened for anti-Salmonella activity. Fifty-eight isolates consisting of Lactobacillus reuteri (n = 32), Lactobacillus salivarius (n = 10), Lactobacillus mucosae (n = 8), Lactobacillus johnsonii (n = 5), and Lactobacillus crispatus (n = 3) were selected and further characterized for probiotic properties including production of antimicrobial substances, acid and bile tolerance, and cell adherence to Caco-2 cells. Eight isolates including Lact. johnsonii LJ202 and Lact. reuteri LR108 were identified as potential probiotics. LJ202 was selected for further use in co-culture studies of two-bacterial and multiple-bacterial species to examine its inhibitory activity against Salmonella enterica serovar Enteritidis DMST7106 (SE7106). Co-culture of LJ202 and SE7106 showed that LJ202 could completely inhibit the growth of SE7106 in 10 h of co-culture. In co-culture of multiple-bacterial species, culturable fecal bacteria from pig feces were used as representative of multiple-bacterial species. The study was performed to examine whether interactions among multiple-bacterial species would influence antagonistic activity of LJ202 against SE7106 and fecal coliform bacteria. Co-culture of SE7106 with different combinations of fecal bacteria and probiotic (LJ202 and LR108) or non-probiotic (Lact. mucosae LM303) strains revealed that the growth of SE7106 was completely inhibited either in the presence or in the absence of probiotic strains. Intriguingly, LJ202 exhibited notable inhibitory activity against fecal coliform bacteria while LR108 did not. Taken together, the results of co-culture studies suggested that LJ202 is a good probiotic candidate for further study its inhibitory effects against pathogen infections in pigs.

  19. Comparing wastewater chemicals, indicator bacteria concentrations, and bacterial pathogen genes as fecal pollution indicators

    USGS Publications Warehouse

    Haack, S.K.; Duris, J.W.; Fogarty, L.R.; Kolpin, D.W.; Focazio, M.J.; Furlong, E.T.; Meyer, M.T.

    2009-01-01

    The objective of this study was to compare fecal indicator bacteria (FIB) (fecal coliforms, Escherichia coli [EC], and enterococci [ENT]) concentrations with a wide array of typical organic wastewater chemicals and selected bacterial genes as indicators of fecal pollution in water samples collected at or near 18 surface water drinking water intakes. Genes tested included esp (indicating human-pathogenic ENT) and nine genes associated with various animal sources of shiga-toxin-producing EC (STEC). Fecal pollution was indicated by genes and/or chemicals for 14 of the 18 tested samples, with little relation to FIB standards. Of 13 samples with <50 EC 100 mL-1, human pharmaceuticals or chemical indicators of wastewater treatment plant effluent occurred in six, veterinary antibiotics were detected in three, and stx1 or stx2 genes (indicating varying animal sources of STEC) were detected in eight. Only the EC eaeA gene was positively correlated with FIB concentrations. Human-source fecal pollution was indicated by the esp gene and the human pharmaceutical carbamazepine in one of the nine samples that met all FIB recreational water quality standards. Escherichia coli rfbO157 and stx2c genes, which are typically associated with cattle sources and are of potential human health significance, were detected in one sample in the absence of tested chemicals. Chemical and gene-based indicators of fecal contamination may be present even when FIB standards are met, and some may, unlike FIB, indicate potential sources. Application of multiple water quality indicators with variable environmental persistence and fate may yield greater confidence in fecal pollution assessment and may inform remediation decisions. Copyright ?? 2009 by the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America. All rights reserved.

  20. Comparing wastewater chemicals, indicator bacteria concentrations, and bacterial pathogen genes as fecal pollution indicators.

    PubMed

    Haack, Sheridan K; Duris, Joseph W; Fogarty, Lisa R; Kolpin, Dana W; Focazio, Michael J; Furlong, Edward T; Meyer, Michael T

    2009-01-01

    The objective of this study was to compare fecal indicator bacteria (FIB) (fecal coliforms, Escherichia coli [EC], and enterococci [ENT]) concentrations with a wide array of typical organic wastewater chemicals and selected bacterial genes as indicators of fecal pollution in water samples collected at or near 18 surface water drinking water intakes. Genes tested included esp (indicating human-pathogenic ENT) and nine genes associated with various animal sources of shiga-toxin-producing EC (STEC). Fecal pollution was indicated by genes and/or chemicals for 14 of the 18 tested samples, with little relation to FIB standards. Of 13 samples with <50 EC 100 mL(-1), human pharmaceuticals or chemical indicators of wastewater treatment plant effluent occurred in six, veterinary antibiotics were detected in three, and stx1 or stx2 genes (indicating varying animal sources of STEC) were detected in eight. Only the EC eaeA gene was positively correlated with FIB concentrations. Human-source fecal pollution was indicated by the esp gene and the human pharmaceutical carbamazepine in one of the nine samples that met all FIB recreational water quality standards. Escherichia coli rfbO157 and stx2c genes, which are typically associated with cattle sources and are of potential human health significance, were detected in one sample in the absence of tested chemicals. Chemical and gene-based indicators of fecal contamination may be present even when FIB standards are met, and some may, unlike FIB, indicate potential sources. Application of multiple water quality indicators with variable environmental persistence and fate may yield greater confidence in fecal pollution assessment and may inform remediation decisions.

  1. The Impact of Rainfall on Fecal Coliform Bacteria in Bayou Dorcheat (North Louisiana)

    PubMed Central

    Hill, Dagne D.; Owens, William E.; Tchounwou, Paul B.

    2006-01-01

    Fecal coliform bacteria are the most common pollutant in rivers and streams. In Louisiana, it has been reported that 37% of surveyed river miles, 31% of lakes, and 23% of estuarine water had some level of contamination. The objective of this research was to assess the effect of surface runoff amounts and rainfall amount parameters on fecal coliform bacterial densities in Bayou Dorcheat in Louisiana. Bayou Dorcheat has been designated by the Louisiana Department of Environmental Quality as a waterway that has uses such as primary contact recreation, secondary contact recreation, propagation of fish and wildlife, agriculture and as being an outstanding natural resource water. Samples from Bayou Dorcheat were collected monthly and analyzed for the presence of fecal coliforms. Fecal coliforms isolated from these samples were identified to the species level. The analysis of the bacterial levels was performed following standard test protocols as described in Standard Methods for the Examination of Water and Wastewater. Information regarding the rainfall amounts and surface runoff amounts for the selected years was retrieved from the Louisiana Office of State Climatology. It was found that a significant increase in the fecal coliform numbers may be associated with average rainfall amounts. Possible sources of elevated coliform counts could include sewage discharges from municipal treatment plants and septic tanks, storm water overflows, and runoff from pastures and range lands. It can be concluded that non-point source pollution that is carried by surface runoff has a significant effect on bacterial levels in water resources. PMID:16823083

  2. Transport of fecal bacteria by boots and vehicle tires in a rural Alaskan community.

    PubMed

    Chambers, Molly K; Ford, Malcolm R; White, Daniel M; Barnes, David L; Schiewer, Silke

    2009-02-01

    People living without piped water and sewer can be at increased risk for diseases transmitted via the fecal-oral route. One rural Alaskan community that relies on hauling water into homes and sewage from homes was studied to determine the pathways of fecal contamination of drinking water and the human environment so that barriers can be established to protect health. Samples were tested for the fecal indicator, Escherichia coli, and the less specific indicator group, total coliforms. Shoes transported fecal contamination from outside to floor material inside buildings. Contamination in puddles on the road, in conjunction with contamination found on all-terrain vehicle (ATV) tires, supports vehicle traffic as a mechanism for transporting contamination from the dumpsite or other source areas to the rest of the community. The abundance of fecal bacteria transported around the community on shoes and ATV tires suggests that centralized measures for waste disposal as well as shoe removal in buildings could improve sanitation and health in the community.

  3. Comparison of fecal indicators with pathogenic bacteria and rotavirus in groundwater

    PubMed Central

    Ferguson, Andrew S.; Layton, Alice C.; Mailloux, Brian J; Culligan, Patricia J.; Williams, Daniel E.; Smartt, Abby E.; Sayler, Gary S.; Feighery, John; McKay, Larry; Knappett, Peter S.K.; Alexandrova, Ekaterina; Arbit, Talia; Emch, Michael; Escamilla, Veronica; Ahmed, Kazi Matin; Alam, Md. Jahangir; Streatfield, P. Kim; Yunus, Mohammad; van Geen, Alexander

    2012-01-01

    Groundwater is routinely analyzed for fecal indicators but direct comparisons of fecal indicators to the presence of bacterial and viral pathogens are rare. This study was conducted in rural Bangladesh where the human population density is high, sanitation is poor, and groundwater pumped from shallow tubewells is often contaminated with fecal bacteria. Five indicator microorganisms (E. coli, total coliform, F+RNA coliphage, Bacteroides and human-associated Bacteroides) and various environmental parameters were compared to the direct detection of waterborne pathogens by quantitative PCR in groundwater pumped from 50 tubewells. Rotavirus was detected in groundwater filtrate from the largest proportion of tubewells (40%), followed by Shigella (10%), Vibrio (10%), and pathogenic E. coli (8%). Spearman rank correlations and sensitivity-specificity calculations indicate that some, but not all, combinations of indicators and environmental parameters can predict the presence of pathogens. Culture-dependent fecal indicator bacteria measured on a single date did not predict total bacterial pathogens, but annually averaged monthly measurements of culturable E. coli did improve prediction for total bacterial pathogens. A qPCR-based E. coli assay was the best indicator for the bacterial pathogens. F+RNA coliphage were neither correlated nor sufficiently sensitive towards rotavirus, but were predictive of bacterial pathogens. Since groundwater cannot be excluded as a significant source of diarrheal disease in Bangladesh and neighboring countries with similar characteristics, the need to develop more effective methods for screening tubewells with respect to microbial contamination is necessary. PMID:22705866

  4. An Assessment of Fecal Coliform Bacteria in Cruise Ship Wastewater Discharge

    DTIC Science & Technology

    2003-09-01

    An Assessment of Fecal Coliform Bacteria in Cruise Ship Wastewater Discharge Charles D. McGee Orange County Sanitation District* 10844 Ellis...Alaska Cruise Ship Initiative in 1999. This initiative required investigation, understanding and oversight of discharges from large cruise ships...into the waters of Alaska. As part of the overall assessment of impacts from cruise ship waste discharges on the environment, a Science Advisory

  5. Isolating the impact of septic systems on fecal pollution in streams of suburban watersheds in Georgia, United States.

    PubMed

    Sowah, Robert A; Habteselassie, Mussie Y; Radcliffe, David E; Bauske, Ellen; Risse, Mark

    2017-01-01

    The presence of multiple sources of fecal pollution at the watershed level presents challenges to efforts aimed at identifying the influence of septic systems. In this study multiple approaches including targeted sampling and monitoring of host-specific Bacteroidales markers were used to identify the impact of septic systems on microbial water quality. Twenty four watersheds with septic density ranging from 8 to 373 septic units/km(2) were monitored for water quality under baseflow conditions over a 3-year period. The levels of the human-associated HF183 marker, as well as total and ruminant Bacteroidales, were quantified using quantitative polymerase chain reaction. Human-associated Bacteroidales yield was significantly higher in high density watersheds compared to low density areas and was negatively correlated (r = -0.64) with the average distance of septic systems to streams in the spring season. The human marker was also positively correlated with the total Bacteroidales marker, suggesting that the human source input was a significant contributor to total fecal pollution in the study area. Multivariable regression analysis indicates that septic systems, along with forest cover, impervious area and specific conductance could explain up to 74% of the variation in human fecal pollution in the spring season. The results suggest septic system impact through contributions to groundwater recharge during baseflow or failing septic system input, especially in areas with >87 septic units/km(2). This study supports the use of microbial source tracking approaches along with traditional fecal indicator bacteria monitoring and land use characterization in a tiered approach to isolate the influence of septic systems on water quality in mixed-use watersheds.

  6. Using host-associated genetic markers to investigate sources of fecal contamination in two Vermont streams

    USGS Publications Warehouse

    Medalie, Laura; Matthews, Leslie J.; Stelzer, Erin A.

    2011-01-01

    , ruminants, or canids. Results from samples from the Mettawee River watershed collected during high-flow conditions (12 storm samples on 2 dates at 6 stations) indicated that there was no evidence of fecal contamination from humans in seven samples and possible evidence in five samples. Results for humans were positive for only one station during both storm events. For two of the five samples with evidence for human fecal contamination, results for two different human genetic markers agreed, but results from three samples were inconsistent. In samples from five of the six Mettawee stations, ruminants were a potential source of fecal contamination on at least one of the three sampled dates, including three positive results for the base-flow sample. Yet samples from all of the stations that showed positive results for ruminants did so for only one or two of the three sampled dates. Samples from only one of the six stations gave consistent results, which were negative for ruminants for all three dates. In the Mettawee River base-flow sample, humans were an unlikely source of major fecal contamination. Factors that may influence results and conclusions include the timing of sample collection relative to the storm event; variability of E. coli and Bacteroidales concentrations in fecal reference samples and in water; sampling and analytical errors; the potential cross reactivity of host-associated genetic markers; and different persistence and survival rates of E. coli bacteria and Bacteroidales genetic markers on land, in water, and by season. These factors interfere with the ability to directly relate Bacteroidales concentrations to E. coli concentrations in river samples. It must be recognized that while use of Bacteroidales genetic markers as a source tracking tool coupled with the interpretive approach described in this report cannot be used quantitatively to pinpoint sources, it can be used to exclude potential sources as major contributors to fecal contamination.

  7. Weather and environmental factors associated with F+ coliphages and fecal indicator bacteria in beach sand at two recreational marine beaches

    EPA Science Inventory

    Studies have demonstrated that fecal indicator bacteria (FIB) and pathogens may be present in beach sand and suggest an increased risk of enteric illness among beachgoers contacting sand. During the 2007 National Epidemiological and Environmental Assessment of Recreational (NEEAR...

  8. Weather and environmental factors associated with F+ coliphages and fecal indicator bacteria in beach sand at two recreational marine beaches

    EPA Science Inventory

    Studies have demonstrated that fecal indicator bacteria (FIB) and pathogens may be present in beach sand and suggest an increased risk of enteric illness among beachgoers contacting sand. During the 2007 National Epidemiological and Environmental Assessment of Recreational (NEEAR...

  9. Effect of solar radiation and predacious microorganisms on survival of fecal and other bacteria.

    PubMed Central

    McCambridge, J; McMeekin, T A

    1981-01-01

    The effect of solar radiation and predacious microorganisms on the survival of bacteria of fecal and plant origin was studied. The decline in the numbers of Escherichia coli cells in estuarine water samples was found to be significantly greater in the presence of both naturally occurring microbial predators and solar radiation than when each of these factors was acting independently. The effect of solar radiation on microbial predators was negligible, whereas the susceptibility of bacteria to light-induced decay varied from one organism to another, as follows: Klebsiella pneumoniae greater than E. coli greater than Salmonella typhimurium, Streptococcus faecium, Enterobacter aerogenes, Erwinia herbicola. PMID:7020590

  10. Vegetable Contamination by the Fecal Bacteria of Poultry Manure: Case Study of Gardening Sites in Southern Benin

    PubMed Central

    Atidégla, Séraphin C.; Huat, Joël; Agbossou, Euloge K.; Saint-Macary, Hervé; Glèlè Kakai, Romain

    2016-01-01

    A study was conducted in southern Benin to assess the contamination of vegetables by fecal coliforms, Escherichia coli, and fecal streptococci as one consequence of the intensification of vegetable cropping through fertilization with poultry manure. For this purpose, on-farm trials were conducted in 2009 and 2010 at Yodo-Condji and Ayi-Guinnou with three replications and four fertilization treatments including poultry manure and three vegetable crops (leafy eggplant, tomato, and carrot). Sampling, laboratory analyses, and counts of fecal bacteria in the samples were performed in different cropping seasons. Whatever the fertilization treatment, the logs of mean fecal bacteria count per g of fresh vegetables were variable but higher than AFNOR criteria. The counts ranged from 8 to 10 fecal coliforms, from 5 to 8 fecal streptococci, and from 2 to 6 Escherichia coli, whereas AFNOR criteria are, respectively, 0, 1, and 0. The long traditional use of poultry manure and its use during the study helped obtain this high population of fecal pathogens. Results confirmed that the contamination of vegetables by fecal bacteria is mainly due to the use of poultry manure. The use of properly composted poultry manure with innovative cropping techniques should help reduce the number and incidence of pathogens. PMID:27069914

  11. Evaluation of Fecal Indicator and Pathogenic Bacteria Originating from Swine Manure Applied to Agricultural Lands Using Culture-Based and Quantitative Real-Time PCR Methods

    EPA Science Inventory

    Fecal bacteria, including those originating from concentrated animal feeding operations, are a leading contributor to water quality impairments in agricultural areas. Rapid and reliable methods are needed that can accurately characterize fecal pollution in agricultural settings....

  12. Evaluation of fecal indicator and pathogenic bacteria originating from swine manure applied to agricultural lands using culture-based and quantitative real-time PCR methods.

    EPA Science Inventory

    Fecal bacteria, including those originating from concentrated animal feeding operations, are a leading contributor to water quality impairments in agricultural areas. Rapid and reliable methods are needed that can accurately characterize fecal pollution in agricultural settings....

  13. Evaluation of fecal indicator and pathogenic bacteria originating from swine manure applied to agricultural lands using culture-based and quantitative real-time PCR methods.

    EPA Science Inventory

    Fecal bacteria, including those originating from concentrated animal feeding operations, are a leading contributor to water quality impairments in agricultural areas. Rapid and reliable methods are needed that can accurately characterize fecal pollution in agricultural settings....

  14. Evaluation of Fecal Indicator and Pathogenic Bacteria Originating from Swine Manure Applied to Agricultural Lands Using Culture-Based and Quantitative Real-Time PCR Methods

    EPA Science Inventory

    Fecal bacteria, including those originating from concentrated animal feeding operations, are a leading contributor to water quality impairments in agricultural areas. Rapid and reliable methods are needed that can accurately characterize fecal pollution in agricultural settings....

  15. Temporal and spatial variability of fecal indicator bacteria in the surf zone off Huntington Beach, CA

    USGS Publications Warehouse

    Rosenfeld, L.K.; McGee, C.D.; Robertson, G.L.; Noble, M.A.; Jones, B.H.

    2006-01-01

    Fecal indicator bacteria concentrations measured in the surf zone off Huntington Beach, CA from July 1998-December 2001 were analyzed with respect to their spatial patterns along 23 km of beach, and temporal variability on time scales from hourly to fortnightly. The majority of samples had bacterial concentrations less than, or equal to, the minimum detection limit, but a small percentage exceeded the California recreational water standards. Areas where coliform bacteria exceeded standards were more prevalent north of the Santa Ana River, whereas enterococci exceedances covered a broad area both north and south of the river. Higher concentrations of bacteria were associated with spring tides. No temporal correspondence was found between these bacterial events and either the timing of cold water pulses near shore due to internal tides, or the presence of southerly swell in the surface wave field. All three fecal indicator bacteria exhibited a diel cycle, but enterococci rebounded to high nighttime values almost as soon as the sun went down, whereas coliform levels were highest near the nighttime low tide, which was also the lower low tide. ?? 2006 Elsevier Ltd. All rights reserved.

  16. Long-Term Survival of Fecal Indicator Bacteria in Estuarine Sediment

    NASA Astrophysics Data System (ADS)

    Ferguson, A. S.; Layton, A.; Culligan, P. J.; Kenna, T. C.; Mailloux, B. J.

    2010-12-01

    Fecal contamination of marine and freshwater environments can negatively impact water quality, leading to contaminated drinking water as well as the closure of recreational beaches and waterways. Fecal contamination is routinely assessed using fecal indicator bacteria (FIB), and even though the potential for their long-term survival or proliferation in sediments exist, information linking deposition of FIB with sediment age is scarce. We evaluate sediments as a reservoir for culturable FIB, by examining dated sediments from the lower Hudson River estuary for Escherichia coli (E. coli), enterococcus, and Bacteroides. Sediment cores were collected from in the vicinity of the George Washington (GWB) and Tappan Zee (TZB) Bridges NY. Sediment deposition ages were constrained using gamma emitting radionuclides and pollution chronology. Culturable E. coli and enterococcus were quantified using a culture-based most probable number method (ColilertTM, Idexx Laboratories). Molecular based methods were used to quantify E. coli and Bacteroides. In the GWB core, viable enterococcus or E. coli were consistently detected in sediment younger than the 1960s with maximum concentrations of 39 and 171 cells/g, respectively. In the TZB core, only enterococcus was sporadically detected in sediment younger than 1950 with a maximum concentration of 79 cells/g. Molecular Bacteroides and E. coli were detected in all core samples with a geometric mean of 4.2x104 and 1.2x105 copies/g, respectively. Results indicate that fecal bacteria can survive within estuarine sediments for decades, suggesting that sediments could be a significant and persistent source of bacterial pollution.

  17. Escherichia coli and fecal-coliform bacteria as indicators of recreational water quality

    USGS Publications Warehouse

    Francy, D.S.; Myers, Donna N.; Metzker, K.D.

    1993-01-01

    In 1986, the U.S. Environmental Protection Agency (USEPA) recommended that Escherichia coli (E. coli) be used in place of fecal-coliform bacteria in State recreational water-quality standards as an indicator of fecal contamination. This announcement followed an epidemiological study in which E. coli concentration was shown to be a better predictor of swimming-associated gastrointestinal illness than fecal-coliform concentration. Water-resource managers from Ohio have decided to collect information specific to their waters and decide whether to use E. coli or fecal-coliform bacteria as the basis for State recreational water-quality standards. If one indicator is a better predictor of recreational water quality than the other and if the relation between the two indicators is variable, then the indicator providing the most accurate measure of recreational water quality should be used in water-quality standards. Water-quality studies of the variability of concentrations of E. coli to fecal-coliform bacteria have shown that (1) concentrations of the two indicators are positively correlated, (2) E. coli to fecal-coliform ratios differ considerably from site to site, and (3) the E. coli criteria recommended by USEPA may be more difficult to meet than current (1992) fecal-coliform standards. In this study, a statistical analysis was done on concentrations of E. coli and fecal-coliform bacteria in water samples collected by two government agencies in Ohio-- the U.S. Geological Survey (USGS) and the Ohio River Valley Water Sanitation Commission (ORSANCO). Data were organized initially into five data sets for statistical analysis: (1) Cuyahoga River, (2) Olentangy River, (3) Scioto River, (4) Ohio River at Anderson Ferry, and (5) Ohio River at Cincinnati Water Works and Tanners Creek. The USGS collected the data in sets 1, 2, and 3, whereas ORSANCO collected the data in sets 4 and 5. The relation of E. coli to fecal-coliform concentration was investigated by use of linear

  18. Fecal-borne bacteria in stormwater and treatment systems in coastal New Hampshire

    NASA Astrophysics Data System (ADS)

    Jones, S. H.

    2005-05-01

    Bacterial contamination is the most common use limitation in New Hampshire's coastal waters. Past studies have shown consistently elevated levels of fecal-borne bacteria in surface waters occur during and following runoff events. Follow-up investigations have shown many stormwater conduits in urban areas that discharge directly into tidal rivers to contain high levels of bacteria, even during dry weather conditions. One of the results of these conditions is the need to close shellfishing waters throughout coastal New Hampshire, especially in Hampton Harbor, following rainfall events. Several recent studies have involved investigation of stormwater treatment system impacts on bacterial pollutants. Influent and effluent water samples from parking lot and storm drain treatment systems, runoff from urban streets, effluent from urban storm drains and receiving water samples were collected during different stages of stormwater runoff and analyzed for fecal coliforms, Escherichia coli and enterococci. E. coli isolates from one storm drain system were also ribotyped to identify source species. The stormwater treatment systems showed different capabilities for removing bacteria. Most were inconsistent at removing bacteria while others showed evidence of possible re-growth of bacteria between storms, especially during warmer weather. Re-growth or illicit connections appear to impact effluent bacterial levels in many urban storm drains. The source species identified for E. coli isolates in one storm drain changed between different stages of a storm event, reflecting runoff dynamics and human behavior patterns. Further work is focused on identifying the most significant sources of bacterial contaminants in receiving waters to help focus ongoing pollution abatement measures.

  19. Fecal-indicator bacteria in the Allegheny, Monongahela, and Ohio Rivers, near Pittsburgh, Pennsylvania, July-September 2001

    USGS Publications Warehouse

    Fulton, John W.; Buckwalter, Theodore F.

    2004-01-01

    This report presents the results of a study by the Allegheny County Health Department (ACHD) and the U.S. Geological Survey (USGS) to determine the concentrations of fecal-indicator bacteria in the Allegheny, Monongahela, and Ohio Rivers (Three Rivers) in Allegheny County, Pittsburgh, Pa. Water-quality samples and river-discharge measurements were collected from July to September 2001 during dry- (72-hour dry antecedent period), mixed-, and wet-weather (48-hour dry antecedent period and at least 0.3 inch of rain in a 6-hour period) conditions at five sampling sites on the Three Rivers in Allegheny County. Water samples were collected weekly to establish baseline conditions and during successive days after three wet-weather events. Water samples were analyzed for fecal-indicator organisms including fecal-coliform (FC) bacteria, Escherichia coli (E. coli), and enterococci bacteria. Water samples were collected by the USGS and analyzed by the ACHD Laboratory. At each site, left-bank and right-bank surface-water samples were collected in addition to a composite sample (discharge-weighted sample representative of the channel cross section as a whole) at each site. Fecal-indicator bacteria reported in bank and composite samples were used to evaluate the distribution and mixing of bacteria-source streams in receiving waters such as the Three Rivers. Single-event concentrations of enterococci, E. coli, and FC during dry-weather events were greater than State and Federal water-quality standards (WQS) in 11, 28, and 28 percent of the samples, respectively; during mixed-weather events, concentrations of fecal-indicator bacteria were greater than WQS in 28, 37, and 43 percent of the samples, respectively; and during wet-weather events, concentrations of fecal-indicator bacteria were greater than WQS in 56, 71, and 81 percent of samples, respectively. Single-event, wet-weather concentrations exceeded those during dry-weather events for all sites except the Allegheny River at

  20. Source tracking fecal bacteria in water: a critical review of current methods.

    PubMed

    Meays, Cynthia L; Broersma, Klaas; Nordin, Rick; Mazumder, Asit

    2004-10-01

    Many molecular and biochemical methods and techniques are being developed to track sources of bacteria in water and food. Currently, there is no standard method proposed for source tracking. This manuscript is a critical evaluation of the various methods used in watersheds, and highlights some of the advantages and disadvantages of each method. Making a decision on a single or combination of methods to use under a particular situation will depend on a number of factors including: question(s) to be answered, scale of identification (broad scale versus specific species identification), available expertise, cost of analysis, turnaround time, and access to facilities. This manuscript reviews several source tracking methodologies which are in current use for source tracking fecal bacteria in the environment including: ribotyping, pulse-field gel electrophoresis, denaturing-gradient gel electrophoresis, repetitive DNA sequences (Rep-PCR), host-specific 16S rDNA genetic markers, and antibiotic resistance analysis.

  1. Modeling Fate and Transport of Fecal Coliform Bacteria Using SWAT 2005 (Case Study: Jajrood River Watershed, Iran)

    NASA Astrophysics Data System (ADS)

    Maghrebi, M.; Tajrishy, M.

    2010-12-01

    Jajrood River watershed is one of the main drinking water resources of the capital city of Tehran, Iran. In addition it has been available as many recreational usages especially in the warm months. As a result of being located near one of the crowded cities of the world, a variety of microbial pollutions is commonly perceived in the Jajrood River. Among them, there are strong concerns about fecal coliform bacteria concentration. This article aimed to model fate and transport of fecal coliform bacteria in Jajrood River watershed using Soil and Water Assessment Tool (SWAT) model version 2005. Potential pollutant sources in the study area were detected and quantified for modeling purposes. In spite of being lack of knowledge about bacteria die-off rate in small river bodies, as well as in other watershed-based forms, fecal coliform bacteria die-off rates were estimated using both laboratory and field data investigations with some simplifications. The SWAT model was calibrated over an extended time period (1997-2002) for this watershed. The river flow calibrated using SUFI-2 software and resulted in a very good outputs (R2=0.82, E=0.81). Furthermore SWAT model was validated over January 2003 to September 2005 in the study area and has resulted in good outputs (R2=0.61, E=0.57). This research illustrates SWAT 2005 capability to model fecal coliform bacteria in a populated watershed, and deals with most of watershed microbial pollution sources that are usually observed in developing countries. Fecal coliform concentration simulation results were mostly in the same order in comparison with real data. However, Differences were judged to be related to lack of input data. In this article different aspects of SWAT capabilities for modeling of fecal coliform bacteria concentration will be reviewed and it will present new insights in bacteria modeling procedures especially for mountainous, high populated and small sized watersheds.

  2. Effects of two diesel fuel mixtures on fecal coliform bacteria densities

    SciTech Connect

    Marcus, J.M.; Scott, G.I.

    1989-03-01

    One of the major potential environmental impacts from synthetic fuel production plants and conventional petroleum refinement operations is the spillage of the refined product into natural waters. Impacts upon aquatic ecosystems resulting from spills of synthetic fuel would likely be different from those associated with conventional petroleum since products extracted from coal or shale are generally richer in phenolics, aromatic amines and other soluble organic compounds. Also, synfuels have higher water solubilities than equivalent petroleum products giving the potential for higher water concentrations of hydrocarbons. This study tested the effects of the water soluble fractions (WSFs) of a shale diesel fuel mixture (SDFM) and a petroleum diesel fuel mixture (PDFM) on the growth of fecal coliform bacteria, the group used almost universally as an indicator of bacteriological water quality. The WSF was tested instead of whole oil because acute toxicity results primarily from this fraction. A wild group of fecal coliform bacteria was used since the objective was to observe effects upon this indicator group encountered in the environment instead of pure laboratory cultures by the routine ambient monitoring and measurement technique of membrane filter colony counts as employed by most water quality management agencies.

  3. Fecal-indicator bacteria in the Allegheny, Monongahela, and Ohio Rivers and selected tributaries, Allegheny County, Pennsylvania, 2001-2005

    USGS Publications Warehouse

    Buckwalter, Theodore F.; Zimmerman, Tammy M.; Fulton, John W.

    2006-01-01

    Concentrations of fecal-indicator bacteria were determined in 1,027 water-quality samples collected from July 2001 through August 2005 during dry- (72-hour dry antecedent period) and wet-weather (48-hour dry antecedent period and at least 0.3 inch of rain in a 24-hour period) conditions in the Allegheny, Monongahela, and Ohio Rivers (locally referred to as the Three Rivers) and selected tributaries in Allegheny County. Samples were collected at five sampling sites on the Three Rivers and at eight sites on four tributaries to the Three Rivers having combined sewer overflows. Water samples were analyzed for three fecal-indicator organisms fecal coliform, Escherichia coli (E. coli), and enterococci bacteria. Left-bank and right-bank surface-water samples were collected in addition to a cross-section composite sample at each site. Concentrations of fecal coliform, E. coli, and enterococci were detected in 98.6, 98.5, and 87.7 percent of all samples, respectively. The maximum fecal-indicator bacteria concentrations were collected from Sawmill Run, a tributary to the Ohio River; Sawmill Run at Duquesne Heights had concentrations of fecal coliform, E. coli, and enterococci of 410,000, 510,000, and 180,000 col/100 mL, respectively, following a large storm. The samples collected in the Three Rivers and selected tributaries frequently exceeded established recreational standards and criteria for bacteria. Concentrations of fecal coliform exceeded the Pennsylvania water-quality standard (200 col/100 mL) in approximately 63 percent of the samples. Sample concentrations of E. coli and enterococci exceeded the U.S. Environmental Protection Agency (USEPA) water-quality criteria (235 and 61 col/100 mL, respectively) in about 53 and 47 percent, respectively, of the samples. Fecal-indicator bacteria were most strongly correlated with streamflow, specific conductance, and turbidity. These correlations most frequently were observed in samples collected from tributary sites. Fecal

  4. Fecal-Indicator Bacteria in the Allegheny, Monongahela, and Ohio Rivers and Selected Tributaries, Allegheny County, Pennsylvania, 2001-2005

    USGS Publications Warehouse

    Buckwalter, Theodore F.; Zimmerman, Tammy M.; Fulton, John W.

    2006-01-01

    Concentrations of fecal-indicator bacteria were determined in 1,027 water-quality samples collected from July 2001 through August 2005 during dry- (72-hour dry antecedent period) and wet-weather (48-hour dry antecedent period and at least 0.3 inch of rain in a 24-hour period) conditions in the Allegheny, Monongahela, and Ohio Rivers (locally referred to as the Three Rivers) and selected tributaries in Allegheny County. Samples were collected at five sampling sites on the Three Rivers and at eight sites on four tributaries to the Three Rivers having combined sewer overflows. Water samples were analyzed for three fecal-indicator organisms fecal coliform, Escherichia coli (E. coli), and enterococci bacteria. Left-bank and right-bank surface-water samples were collected in addition to a cross-section composite sample at each site. Concentrations of fecal coliform, E. coli, and enterococci were detected in 98.6, 98.5, and 87.7 percent of all samples, respectively. The maximum fecal-indicator bacteria concentrations were collected from Sawmill Run, a tributary to the Ohio River; Sawmill Run at Duquesne Heights had concentrations of fecal coliform, E. coli, and enterococci of 410,000, 510,000, and 180,000 col/100 mL, respectively, following a large storm. The samples collected in the Three Rivers and selected tributaries frequently exceeded established recreational standards and criteria for bacteria. Concentrations of fecal coliform exceeded the Pennsylvania water-quality standard (200 col/100 mL) in approximately 63 percent of the samples. Sample concentrations of E. coli and enterococci exceeded the U.S. Environmental Protection Agency (USEPA) water-quality criteria (235 and 61 col/100 mL, respectively) in about 53 and 47 percent, respectively, of the samples. Fecal-indicator bacteria were most strongly correlated with streamflow, specific conductance, and turbidity. These correlations most frequently were observed in samples collected from tributary sites. Fecal

  5. Effect of external or internal fecal contamination on numbers of bacteria on prechilled broiler carcasses.

    PubMed

    Smith, D P; Northcutt, J K; Cason, J A; Hinton, A; Buhr, R J; Ingram, K D

    2007-06-01

    During processing, fecal material may contact broiler carcasses externally or internally. A study was conducted to determine the effect of external vs. internal fecal contamination on numbers of bacteria on broiler carcasses. In each of 3 trials, 12 carcasses just prior to evisceration were obtained from a commercial processing plant, placed on a shackle line, and eviscerated with commercial equipment in a pilot scale processing plant. Also, approximately 20 intestinal tracts were collected from the processing plant; then cecal contents were collected and pooled. One gram of cecal content was placed on the exterior breast skin (external), inside the carcass cavity (internal), or not applied (control). All carcasses were held 10 min, then placed on the shackle line and passed through a commercial inside-outside bird washer set at 552 kPa, 5 s dwell time, using approximately 189 L per min of tap water at ambient temperature. After a 1-min drip, whole carcass rinses were conducted on each carcass, and coliforms, Escherichia coli, and Campylobacter counts were determined and reported as log cfu/mL of rinse. External carcass contamination resulted in significantly higher (P<0.05) coliform, E. coli, and Campylobacter numbers than internal contamination (5.0 vs. 4.5, 4.9 vs. 4.2, and 3.6 vs. 2.6, respectively). Control carcass counts were significantly lower than external or internal carcass contamination counts for coliforms (3.7), E. coli (3.6), and Campylobacter (2.2). External contamination resulted in higher numbers of bacteria after carcass washing, but carcasses with internal contamination still have higher numbers of bacteria after washing than carcasses without applied contamination.

  6. Movement of traditional fecal indicator bacteria and source-tracking targets through septic drainfields.

    PubMed

    Billian, Hannah; Krometis, Leigh-Anne; Thompson, Theresa; Hagedorn, Charles

    2018-01-01

    The past three decades' data on outbreaks in the United States indicate that homes dependent on untreated groundwater (e.g. wells) for household drinking water that are also reliant on onsite treatment of household wastewater (e.g. septic systems) may be at greater risk for waterborne disease. While groundwater quality monitoring to protect public health has traditionally focused on the detection of fecal indicator bacteria, the application of emerging source tracking strategies may offer a more efficient means to identify pollution sources and effective means of remediation. This study compares the movement of common fecal indicator bacteria (E. coli and enterococci) with a chemical (optical brighteners, OB) and a molecular (Bacteroides HF183) source tracking (ST) target in small scale septic drainfield models in order to evaluate their potential utility in groundwater monitoring. Nine PVC column drainfield models received synchronized doses of primary-treated wastewater twice daily, with influent and effluent monitored bi-weekly over a 7-month period for all targets. Results indicate that E. coli and enterococci concentrations were strongly associated (Spearman's rank, p<0.05), and correlations between enterococci and optical brighteners were moderately strong. Bacteroides HF183 was significantly, but not strongly, associated with optical brighteners and both indicator bacteria (Point-biserial correlation, p<0.05), most likely due to its sporadic detection. Application of human ST marker monitoring in groundwaters at risk of contamination by human sewage is recommended, although consistent interpretation of results will rely on more detailed evaluation of HF183 incidence in source contamination waters. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. The effect of storage time on Vibrio spp. and fecal indicator bacteria in an Isco autosampler.

    PubMed

    Ghazaleh, Maite N; Froelich, Brett A; Noble, Rachel T

    2014-09-01

    Monitoring concentrations of bacterial pathogens and indicators of fecal contamination in coastal and estuarine ecosystems is critical to reduce adverse effects to public health. During storm events, particularly hurricanes, floods, Nor'easters, and tropical cyclones, sampling of coastal and estuarine waters is not generally possible due to safety concerns. It is particularly important to monitor waters during these periods as it is at precisely these times that pathogenic bacteria such as Vibrio spp. and fecal indicator bacteria concentrations fluctuate, potentially posing significant risks to public health. Automated samplers, such as the Isco sampler, are commonly used to conduct remote sample collection. Remote sampling is employed during severe storm periods, thereby reducing risk to researchers. Water samples are then stored until conditions are safe enough to retrieve them, typically in less than 21h, to collect the samples. Concerns exist regarding potential "bottle effects", whereby containment of sample might result in altered results. While these effects are well documented in samples being held for 24h or more, there is little data on bottle effects occurring during the first 24h of containment, and less still on the specific effects related to this type of sampling regime. Estuarine water samples were collected in the fall of 2013, placed into an Isco autosampler and subsampled over time to determine the effects of storage within this type of autosampling device. Vibrio spp. and fecal indicator bacteria were quantified using replicated culture-based methods, including Enterolert™ and membrane filtration. The experiments demonstrated no significant impact of storage time when comparing concentrations of total Vibrio spp., Vibrio vulnificus, Vibrio parahaemolyticus, or Enterococcus spp. after storage compared to original concentrations. However, the findings also suggested that increased variability and growth can occur during the middle of the day

  8. Fecal indicator bacteria persistence under natural conditions in an ice-covered river.

    PubMed Central

    Davenport, C V; Sparrow, E B; Gordon, R C

    1976-01-01

    Total coliform (TC), fecal coliform (FC), and fecal streptococcus (FS) survival characteristics, under natural conditions at 0 degrees C in an ice-covered river, were examined during February and March 1975. The membrane filter (MF) technique was used throughout the study, and the multiple-tube (MPN) method was used in parallel on three preselected days for comparative recovery of these bacteria. Survival was studied at seven sample stations downstream from all domestic pollution sources in a 317-km reach of the river having 7.1 days mean flow time (range of 6.0 to 9.1 days). The mean indicator bacteria densities decreased continuously at successive stations in this reach and, after adjustment for dilution, the most rapid die-off was found to occur during the first 1.9 days, followed by a slower decrease. After 7.1 days, the relative survival was TC less than FC less than FS, with 8.4%, 15.7%, and 32.8% of the initial populations remaining viable, respectively. These rates are higher than previously reported and suggest that the highest survival rates for these bacteria in receiving streams can be expected at 0 degree C under ice cover. Additionally, the FC-FS ratio was greater than 5 at all stations, indicating that this ratio may be useable for determining the source of fecal pollution in receiving streams for greater than 7 days flow time at low water temperatures. The MPN and MF methods gave comparable results for the TC and FS at all seven sample stations, with both the direct and verified MF counts within the 95% confidence limits of the respective MPNs in most samples, but generally lower than the MPN index. Although FC recovery on membrane filters was comparable results at stations near the pollution source. However, the results became more comparable with increasing flow time. The results of this study indicate that heat shock is a major factor in suppression of the FC counts on the membrane filters at 44.5 degree C. Heat shock may be minimized by extended

  9. Presence and Sources of Fecal Coliform Bacteria in Epilithic Periphyton Communities of Lake Superior▿

    PubMed Central

    Ksoll, Winfried B.; Ishii, Satoshi; Sadowsky, Michael J.; Hicks, Randall E.

    2007-01-01

    Epilithic periphyton communities were sampled at three sites on the Minnesota shoreline of Lake Superior from June 2004 to August 2005 to determine if fecal coliforms and Escherichia coli were present throughout the ice-free season. Fecal coliform densities increased up to 4 orders of magnitude in early summer, reached peaks of up to 1.4 × 105 CFU cm−2 by late July, and decreased during autumn. Horizontal, fluorophore-enhanced repetitive-PCR DNA fingerprint analyses indicated that the source for 2% to 44% of the E. coli bacteria isolated from these periphyton communities could be identified when compared with a library of E. coli fingerprints from animal hosts and sewage. Waterfowl were the major source (68 to 99%) of periphyton E. coli strains that could be identified. Several periphyton E. coli isolates were genotypically identical (≥92% similarity), repeatedly isolated over time, and unidentified when compared to the source library, suggesting that these strains were naturalized members of periphyton communities. If the unidentified E. coli strains from periphyton were added to the known source library, then 57% to 81% of E. coli strains from overlying waters could be identified, with waterfowl (15 to 67%), periphyton (6 to 28%), and sewage effluent (8 to 28%) being the major potential sources. Inoculated E. coli rapidly colonized natural periphyton in laboratory microcosms and persisted for several weeks, and some cells were released to the overlying water. Our results indicate that E. coli from periphyton released into waterways confounds the use of this bacterium as a reliable indicator of recent fecal pollution. PMID:17468280

  10. Distribution and variability of fecal-indicator bacteria in Scioto and Olentangy rivers in the Columbus, Ohio, area

    USGS Publications Warehouse

    Myers, Donna N.

    1992-01-01

    This report presents the results of a study by the U.S. Geological Survey, in cooperation with the City of Columbus, Ohio, to determine the distribution and variability of fecal-indicator bacteria in Scioto and Olentangy Rivers. Fecal-indicator bacteria are among the contaminants of concern to recreational users of these rivers in the Columbus area. Samples were collected to be analyzed for fecal-coliform and Escherichia coli (E. coli) bacteria and selected water-quality constituents and physical properties at 10 sites-- 4 on the Olentangy River and 6 on the Scioto River during the recreational seasons in 1987, 1988, and 1989. Measurements of streamflow also were made at these sites at various frequencies during base flow and runoff. The concentrations of fecal-coliform and E. coli bacteria in the Scioto and Olentangy Rivers spanned a range of five orders of magnitude, from less than 20 to greater than 2,000,000 col/100 mL (colonies per 100 milliliters). In addition, the concentrations of fecal coliform and E. coli bacteria are well correlated (r=0.97) in the study area. At times, relatively high concentrations, for fecal-indicator bacteria (concentrations greater than 51,000 col/100 mL for fecal-coliform and E. coli ) were found in Olentangy River at Woody Hayes Drive and at Goodale Street, and in Scioto River at Greenlawn Avenue and at Columbus. Intermediate concentrations of fecal-indicator bacteria (from 5,100 to 50,000 col/100 mL for fecal coliform and (from 510 to 50,000 col/100 mL for E. coli ) were found in Scioto River at Town Street and below O'Shaughnessy Dam near Dublin, Ohio, and in Olentangy River at Henderson Road. The lowest (median) concentrations of fecal-indicator bacteria (from 20 to 5,000 col/100 mL for fecal coliform and from 20 to 500 col/100 mL for E. coli ) were found at Olentangy River near Worthington, Ohio, Scioto River at Dublin Road Water Treatment Plant and below Griggs Reservoir. Fecal-coliform concentrations exceeded the geometric

  11. Influence of manure age and sunlight on the community structure of cattle fecal bacteria as revealed by Illumina sequencing

    NASA Astrophysics Data System (ADS)

    Wong, K.; Shaw, T. I.; Oladeinde, A.; Molina, M.

    2013-12-01

    Fecal pollution of environmental waters is a major concern for the general public because exposure to fecal-associated pathogens can have severe impacts on human health. Stream and river impairment due to fecal pollution is largely the result of agricultural activities in the United States. In the last few years, numerous metagenomic studies utilized next generation sequencing to develop microbial community profiles by massively sequencing the 16sRNA hypervariable region. This technology supports the application of water quality assessment such as pathogen detection and fecal source tracking. The bacteria communities of samples in these studies were determined when they were freshly collected; therefore, little is known about how feces age or how environmental stress influences the microbial ecology of fecal materials. In this study we monitored bacteria community changes in cattle feces for 57 days after excretion (day 0, 2, 4 8, 15, 22, 29, 43, 57) by sequencing the 16s variable region 4, using Illumnia MiSeq. Twelve cattle feces were studied; half of the samples were directly exposed to sunlight (unshaded) and half were shaded. Results indicate that the relative abundance (RA) profile in both shaded and unshaded samples rapidly changed from day 0 to 15, but stabilized from day 22 to 57. Firmcutes were the most abundant phylum (~40%) at day 0, but were reduced to <10% by day 57. The RA of Proteobacteria was only 1% at day 0, but increased to ~50% by day 57in both shaded and unshaded samples. By the end of the study, shaded and unshaded samples had a similar RA of Firmcutes and Proteobacteria but the RA of Bacteroidetes and Actinobacteria was, respectively, about 7% lower and 10% higher for unshaded samples. UV intensity, moisture, and temperature were significantly different between shaded and unshaded plots, indicating that these environmental stresses could influence the structure of fecal bacteria community in the natural environment. According to the

  12. Stochastic Analysis of Non Point Source Loading of Fecal Bacteria in a Shallow Heterogeneous Aquifer

    NASA Astrophysics Data System (ADS)

    Cook, S. J.; Li, X.; Atwill, R.; Packman, A. I.; Harter, T.

    2011-12-01

    Manure and wastewater irrigation (MWI) presents a microbiological risk to shallow groundwater quality. Particularly vulnerable are domestic wells in rural areas where treatment systems may be limited or unreliable. However, despite multiple and persistent sources of fecal contamination, cross sectional monitoring of fecal bacteria in groundwater indicates a high degree of variability in both prevalence and measured concentrations. Apparently random variation occurs both between wells and samples at individual wells. In contrast, deliberate longitudinal studies of MWIs, particularly in the laboratory, tend to exhibit relatively smooth breakthrough curves consistent with colloid filtration theory. To better characterize potential sources of variability in observed field data, a 3D stochastic groundwater modeling approach representative of irrigation applications to vulnerable alluvial aquifers was developed. Heterogeniety is assessed by incorporating multiple loading functions and hydrostratigraphic representations of a heterogeneous alluvial aquifer. Simulations indicate that irrigation water breakthroughs to wells are generally limited to shallow depths, suggesting limited risk to domestic wells screened several tens of meters below the water table. Whilst the presence of aquifer heterogeneity significantly extends the transport distance and tailing of breakthrough curves, owing to macro-dispersion and in-well mixing, simulated breakthrough curves are relatively smooth and consistent with observed longitudinal studies. This suggests that the highly erratic and variable nature of microorganism detection may be due to highly transient processes, including but not limited to spatio-temporal variations in source variability and limitations in infrequent monitoring programs to properly determine variability.

  13. Particle-size distribution as indicator for fecal bacteria contamination of drinking water from karst springs.

    PubMed

    Pronk, Michiel; Goldscheider, Nico; Zopfi, Jakob

    2007-12-15

    Continuous monitoring of particle-size distribution (PSD), total organic carbon (TOC), turbidity, discharge and physicochemical parameters, together with analyses of fecal indicator bacteria, particularly Escherichia coli, made it possible to better understand the processes governing pathogen transport in karst groundwater and to establish PSD as indicator for possible microbial contamination of drinking water from karst springs. In the study area near Yverdon-les-Bains, Switzerland, tracer tests proved connection between a sinking stream draining agricultural land and several springs, 4.8-6.3 km away. Tracing and monitoring results demonstrate that (i) suspended particles (turbidity) in the spring water either originate from remobilization of sediments inside the aquifer (autochthonous) or from the sinking stream and land surface (allochthonous); (ii) allochthonous turbidity coincides with increased E. coli and TOC levels; (iii) PSD makes it possible to distinguish the two types of turbidity; (iv) a relative increase of finer particles (0.9-10 microm) indicates allochthonous turbidity and thus possible fecal contamination. The method permits to optimize water treatment and identify periods when the spring water must be rejected. Findings from other test sites confirm the feasibility of this approach.

  14. Cultivation-Independent Analysis of Bacteria in IDEXX Quanti-Tray/2000 Fecal Indicator Assays ▿

    PubMed Central

    Sercu, Bram; Van De Werfhorst, Laurie C.; Murray, Jill L. S.; Holden, Patricia A.

    2011-01-01

    Monitoring microbiological water quality is important for protecting water resources and the health of swimmers. Routine monitoring relies on cultivating fecal indicator bacteria (FIB), frequently using defined substrate technology. Defined substrate technology is designed to specifically enrich for FIB, but a complete understanding of the assay microbiology requires culture-independent analysis of the enrichments. This study aimed to identify bacteria in positive wells of Colilert and Enterolert Quanti-Tray/2000 (IDEXX Laboratories) FIB assays in environmental water samples and to quantify the degree of false-positive results for samples from an urban creek by molecular methods. Pooled Escherichia coli- and Enterococcus-positive Quanti-Tray/2000 enrichments, either from urban creek dry weather flow or municipal sewage, harbored diverse bacterial populations based on 16S rRNA gene sequences and terminal restriction fragment length polymorphism analyses. Target taxa (coliforms or enterococci) and nontarget taxa (Vibrio spp., Shewanella spp., Bacteroidetes, and Clostridium spp.) were identified in pooled and individual positive Colilert and Enterolert wells based on terminal restriction fragments that were in common with those generated in silico from clone sequences. False-positive rates of between 4 and 23% occurred for the urban creek samples, based on the absence of target terminal restriction fragments in individual positive wells. This study suggests that increased selective inhibition of nontarget bacteria could improve the accuracy of the Colilert and Enterolert assays. PMID:21097584

  15. A short review of fecal indicator bacteria in tropical aquatic ecosystems: knowledge gaps and future directions

    PubMed Central

    Rochelle-Newall, Emma; Nguyen, Thi Mai Huong; Le, Thi Phuong Quynh; Sengtaheuanghoung, Oloth; Ribolzi, Olivier

    2015-01-01

    Given the high numbers of deaths and the debilitating nature of diseases caused by the use of unclean water it is imperative that we have an understanding of the factors that control the dispersion of water borne pathogens and their respective indicators. This is all the more important in developing countries where significant proportions of the population often have little or no access to clean drinking water supplies. Moreover, and notwithstanding the importance of these bacteria in terms of public health, at present little work exists on the persistence, transfer and proliferation of these pathogens and their respective indicator organisms, e.g., fecal indicator bacteria (FIB) such as Escherichia coli and fecal coliforms in humid tropical systems, such as are found in South East Asia or in the tropical regions of Africa. Both FIB and the waterborne pathogens they are supposed to indicate are particularly susceptible to shifts in water flow and quality and the predicted increases in rainfall and floods due to climate change will only exacerbate the problems of contamination. This will be furthermore compounded by the increasing urbanization and agricultural intensification that developing regions are experiencing. Therefore, recognizing and understanding the link between human activities, natural process and microbial functioning and their ultimate impacts on human health are prerequisites for reducing the risks to the exposed populations. Most of the existing work in tropical systems has been based on the application of temperate indicator organisms, models and mechanisms regardless of their applicability or appropriateness for tropical environments. Here, we present a short review on the factors that control FIB dynamics in temperate systems and discuss their applicability to tropical environments. We then highlight some of the knowledge gaps in order to stimulate future research in this field in the tropics. PMID:25941519

  16. [Preliminary study on bacteroides as the potential fecal contamination indicator bacteria].

    PubMed

    Yang, Jing-yan; Chen, Zhi-jin; Ding, Xiao-bei; Huang, Wei; Yang, Rui-jia; Pei, Xiao-fang

    2011-03-01

    To explore the possibility of Bacteroides spp. as fecal contamination indicator bacteria with real-time quantitative PCR (RT-PCR) assay through analyzing the correlation between Bacteroides spp. and coliform group in external environment. Quantity of coliform group and Bacteroides in water samples were detected by most-probable-number method (MPN) and RT-PCR, respectively, and their detection correlation was evaluated with linear correlation analysis. Both methods were also applied to detect the contaminated time limits and river water samples collected at four sampling sites in three different times. Seventy two hours were needed for the numeration of coliform group with MPN method, while RT-PCR could detect Bacteroides within 3 hours. The contaminated time limit of indoor and outdoor water samples of coliform group was more than 40 days and 9 days, and Bacteroides 13 days and 5 days, respectively. Also, the positive correlation between the quantity of Bacteroides and coliform group in outdoor water samples was obtained, the quantity of Bacteroides was from 8.3 × 10(6) copies/ml to less than 10(4) copies/ml during the first day to the fifth day, while coliform group was 4.3 × 10(6) MPN/100 ml to 2.4 × 10(3) MPN/100 ml. A 100% coincidence rate of the detection results with both methods was also observed. These results indicated that the detection results of both methods had perfect consistency. Bacteroides spp. can be potentially used as fecal contamination indicator bacteria with RT-PCR rapid detection.

  17. Bayesian meta-analysis to synthesize decay rate constant estimates for common fecal indicator bacteria.

    PubMed

    Brooks, Lauren E; Field, Katharine G

    2016-11-01

    For decades, fecal indicator bacteria have been used as proxies to quantitatively estimate fecal loading into water bodies. Widely used cultured indicators (e.g. Escherichia coli and Enterococcus spp.) and more recently developed genetic markers are well studied, but their decay in the environment is still poorly understood. We used Hierarchical Bayesian Linear Modeling to conduct a series of meta-analyses using published decay rate constant estimates, to synthesize findings into pooled estimates and identify gaps in the data preventing reliable estimates. In addition to the meta-analysis assuming all estimates come from the same population, meta-regressions including covariates believed to contribute to decay were fit and used to provided synthesized estimates for specific combinations of significant variables. Additionally, statements regarding the significance of variables across studies were made using the 95% confidence interval for meta-regression coefficients. These models were used to construct a mean decay rate constant estimate as well as credible intervals for the mean and the distribution of all likely data points. While synthesized estimates for each targeted indicator bacteria were developed, the amount of data available varied widely for each target, as did the predictive power of the models as determined by testing with additional data not included in the modeling. Temperature was found to be significant for all selected indicators, while light was found to be significant only for culturable indicators. Results from the models must be interpreted with caution, as they are based only on the data available, which may not be representative of decay in other scenarios. Copyright © 2016 Elsevier Ltd. All rights reserved.

  18. Ruminants Contribute Fecal Contamination to the Urban Household Environment in Dhaka, Bangladesh.

    PubMed

    Harris, Angela R; Pickering, Amy J; Harris, Michael; Doza, Solaiman; Islam, M Sirajul; Unicomb, Leanne; Luby, Stephen; Davis, Jennifer; Boehm, Alexandria B

    2016-05-03

    In Dhaka, Bangladesh, the sensitivity and specificity of three human, three ruminant, and one avian source-associated QPCR microbial source tracking assays were evaluated using fecal samples collected on site. Ruminant-associated assays performed well, whereas the avian and human assays exhibited unacceptable cross-reactions with feces from other hosts. Subsequently, child hand rinses (n = 44) and floor sponge samples (n = 44) from low-income-households in Dhaka were assayed for fecal indicator bacteria (enterococci, Bacteroidales, and Escherichia coli) and a ruminant-associated bacterial target (BacR). Mean enterococci concentrations were of 100 most probable number (MPN)/2 hands and 1000 MPN/225 cm(2) floor. Mean concentrations of Bacteroidales were 10(6) copies/2 hands and 10(5) copies/225 cm(2) floor. E. coli were detected in a quarter of hand rinse and floor samples. BacR was detected in 18% of hand rinse and 27% of floor samples. Results suggest that effective household fecal management should account not only for human sources of contamination but also for animal sources. The poor performance of the human-associated assays in the study area calls into the question the feasibility of developing a human-associated marker in urban slum environments, where domestic animals are exposed to human feces that have been disposed in pits and open drains.

  19. Fecal-indicator bacteria concentrations in the Illinois River between Hennepin and Peoria, Illinois: 2007-08

    USGS Publications Warehouse

    Dupre, David H.; Hortness, Jon E.; Terrio, Paul J.; Sharpe, Jennifer B.

    2012-01-01

    The Illinois Environmental Protection Agency has designated portions of the Illinois River in Peoria, Woodford, and Tazewell Counties, Illinois, as impaired owing to the presence of fecal coliform bacteria. The U.S. Geological Survey, in cooperation with the Tri-County Regional Planning Commission, examined the water quality in the Illinois River and major tributaries within a 47-mile reach between Peoria and Hennepin, Ill., during water year 2008 (October 2007–September 2008). Investigations included synoptic (snapshot) sampling at multiple locations in a 1-day period: once in October 2007 during lower streamflow conditions, and again in June 2008 during higher streamflow conditions. Five locations in the study area were monitored for the entire year at monthly or more frequent intervals. Two indicator bacteria were analyzed in each water sample: fecal coliform and Escherichia coli (E. coli). Streamflow information from previously established monitoring locations in the study area was used in the analysis. Correlation analyses were used to characterize the relation between the two fecal-indicator bacteria and the relation of either indicator to streamflow. Concentrations of the two measured fecal-indicator bacteria correlated well for all samples analyzed (r = 0.94, p E. coli: rho = -0.43, p = 0.0157). The correlation between fecal indicators and streamflow in tributaries or in the Illinois River at Hennepin was found to be statistically significant, yet moderate in strength with coefficient values ranging from r = 0.4 to 0.6. Indirect observations from the June 2008 higher flow synoptic event may indicate continued effects from combined storm and sanitary sewers in the vicinity of the Illinois River near Peoria, Ill., contributing to observed single-sample exceedance of the State criterion for fecal coliform.

  20. Inter-laboratory Comparison of Real-time PCR Methods for Quantification of General Fecal Indicator Bacteria

    EPA Science Inventory

    The application of quantitative real-time PCR (qPCR) technologies for the rapid identification of fecal bacteria in environmental waters is being considered for use as a national water quality metric in the United States. The transition from research tool to a standardized prot...

  1. Assessing Environmental Impacts of Treated Wastewater through Monitoring of Fecal Indicator Bacteria and Salinity in Irrigated Soils

    USDA-ARS?s Scientific Manuscript database

    To assess the potential for bacterial persistence and/or growth in reclaimed water irrigation systems and in irrigated soils, and to quantify the effects of wastewater application on soil salinity, levels of fecal indicator bacteria (E. coli, Enterococcus) and environmental covariates were measured ...

  2. Estimation of decay rates for fecal indicator bacteria and bacterial pathogens in agricultural field-applied manure

    EPA Science Inventory

    Field-applied manure is an important source of pathogenic exposure in surface water bodies for humans and ecological receptors. We analyzed the persistence and decay of fecal indicator bacteria and bacterial pathogens from three sources (cattle, poultry, swine) for agricultural f...

  3. COMPARISON OF REAL-TIME PCR FECAL BACTERIA MEASUREMENTS IN RECREATIONAL WATERS USING DIFFERENT INSTRUMENTS AND REAGENT SYSTEMS

    EPA Science Inventory

    U.S. EPA guidance on the safety of surface waters for recreational use is currently based on concentrations of culturable fecal indicator bacteria. Attention is now shifting to more rapid molecular monitoring methods. A multi-year epidemiological study is in progress to determine...

  4. Inter-laboratory Comparison of Real-time PCR Methods for Quantification of General Fecal Indicator Bacteria

    EPA Science Inventory

    The application of quantitative real-time PCR (qPCR) technologies for the rapid identification of fecal bacteria in environmental waters is being considered for use as a national water quality metric in the United States. The transition from research tool to a standardized prot...

  5. Estimation of decay rates for fecal indicator bacteria and bacterial pathogens in agricultural field-applied manure

    EPA Science Inventory

    Field-applied manure is an important source of pathogenic exposure in surface water bodies for humans and ecological receptors. We analyzed the persistence and decay of fecal indicator bacteria and bacterial pathogens from three sources (cattle, poultry, swine) for agricultural f...

  6. COMPARISON OF REAL-TIME PCR FECAL BACTERIA MEASUREMENTS IN RECREATIONAL WATERS USING DIFFERENT INSTRUMENTS AND REAGENT SYSTEMS

    EPA Science Inventory

    U.S. EPA guidance on the safety of surface waters for recreational use is currently based on concentrations of culturable fecal indicator bacteria. Attention is now shifting to more rapid molecular monitoring methods. A multi-year epidemiological study is in progress to determine...

  7. Differential Decay of Cattle-associated Fecal Indicator Bacteria and Microbial Source Tracking Markers in Fresh and Marine Water

    EPA Science Inventory

    Background: Fecal indicator bacteria (FIB) have a long history of use in the assessment of the microbial quality of recreational waters. However, quantification of FIB provides no information about the pollution source(s) and relatively little is known about their fate in the amb...

  8. Partial least squares for efficient models of fecal indicator bacteria on Great Lakes beaches

    USGS Publications Warehouse

    Brooks, Wesley R.; Fienen, Michael N.; Corsi, Steven R.

    2013-01-01

    At public beaches, it is now common to mitigate the impact of water-borne pathogens by posting a swimmer's advisory when the concentration of fecal indicator bacteria (FIB) exceeds an action threshold. Since culturing the bacteria delays public notification when dangerous conditions exist, regression models are sometimes used to predict the FIB concentration based on readily-available environmental measurements. It is hard to know which environmental parameters are relevant to predicting FIB concentration, and the parameters are usually correlated, which can hurt the predictive power of a regression model. Here the method of partial least squares (PLS) is introduced to automate the regression modeling process. Model selection is reduced to the process of setting a tuning parameter to control the decision threshold that separates predicted exceedances of the standard from predicted non-exceedances. The method is validated by application to four Great Lakes beaches during the summer of 2010. Performance of the PLS models compares favorably to that of the existing state-of-the-art regression models at these four sites.

  9. Climate change and land use drivers of fecal bacteria in tropical hawaiian rivers.

    PubMed

    Strauch, Ayron M; Mackenzie, Richard A; Bruland, Gregory L; Tingley, Ralph; Giardina, Christian P

    2014-07-01

    Potential shifts in rainfall driven by climate change are anticipated to affect watershed processes (e.g., soil moisture, runoff, stream flow), yet few model systems exist in the tropics to test hypotheses about how these processes may respond to these shifts. We used a sequence of nine watersheds on Hawaii Island spanning 3000 mm (7500-4500 mm) of mean annual rainfall (MAR) to investigate the effects of short-term (24-h) and long-term (MAR) rainfall on three fecal indicator bacteria (FIB) (enterococci, total coliforms, and ). All sample sites were in native Ohia dominated forest above 600 m in elevation. Additional samples were collected just above sea level where the predominant land cover is pasture and agriculture, permitting the additional study of interactions between land use across the MAR gradient. We found that declines in MAR significantly amplified concentrations of all three FIB and that FIB yield increased more rapidly with 24-h rainfall in low-MAR watersheds than in high-MAR watersheds. Because storm frequency decreases with declining MAR, the rate of change in water potential affects microbial growth, whereas increased rainfall intensity dislodges more soil and bacteria as runoff compared with water-logged soils of high-MAR watersheds. As expected, declines in % forest cover and increased urbanization increased FIB. Taken together, shifts in rainfall may alter bacterial inputs to tropical streams, with land use change also affecting water quality in streams and near-shore environments.

  10. Classification of antibiotic resistance patterns of indicator bacteria by discriminant analysis: use in predicting the source of fecal contamination in subtropical waters.

    PubMed

    Harwood, V J; Whitlock, J; Withington, V

    2000-09-01

    The antibiotic resistance patterns of fecal streptococci and fecal coliforms isolated from domestic wastewater and animal feces were determined using a battery of antibiotics (amoxicillin, ampicillin, cephalothin, chlortetracycline, oxytetracycline, tetracycline, erythromycin, streptomycin, and vancomycin) at four concentrations each. The sources of animal feces included wild birds, cattle, chickens, dogs, pigs, and raccoons. Antibiotic resistance patterns of fecal streptococci and fecal coliforms from known sources were grouped into two separate databases, and discriminant analysis of these patterns was used to establish the relationship between the antibiotic resistance patterns and the bacterial source. The fecal streptococcus and fecal coliform databases classified isolates from known sources with similar accuracies. The average rate of correct classification for the fecal streptococcus database was 62.3%, and that for the fecal coliform database was 63.9%. The sources of fecal streptococci and fecal coliforms isolated from surface waters were identified by discriminant analysis of their antibiotic resistance patterns. Both databases identified the source of indicator bacteria isolated from surface waters directly impacted by septic tank discharges as human. At sample sites selected for relatively low anthropogenic impact, the dominant sources of indicator bacteria were identified as various animals. The antibiotic resistance analysis technique promises to be a useful tool in assessing sources of fecal contamination in subtropical waters, such as those in Florida.

  11. Classification of Antibiotic Resistance Patterns of Indicator Bacteria by Discriminant Analysis: Use in Predicting the Source of Fecal Contamination in Subtropical Waters

    PubMed Central

    Harwood, Valerie J.; Whitlock, John; Withington, Victoria

    2000-01-01

    The antibiotic resistance patterns of fecal streptococci and fecal coliforms isolated from domestic wastewater and animal feces were determined using a battery of antibiotics (amoxicillin, ampicillin, cephalothin, chlortetracycline, oxytetracycline, tetracycline, erythromycin, streptomycin, and vancomycin) at four concentrations each. The sources of animal feces included wild birds, cattle, chickens, dogs, pigs, and raccoons. Antibiotic resistance patterns of fecal streptococci and fecal coliforms from known sources were grouped into two separate databases, and discriminant analysis of these patterns was used to establish the relationship between the antibiotic resistance patterns and the bacterial source. The fecal streptococcus and fecal coliform databases classified isolates from known sources with similar accuracies. The average rate of correct classification for the fecal streptococcus database was 62.3%, and that for the fecal coliform database was 63.9%. The sources of fecal streptococci and fecal coliforms isolated from surface waters were identified by discriminant analysis of their antibiotic resistance patterns. Both databases identified the source of indicator bacteria isolated from surface waters directly impacted by septic tank discharges as human. At sample sites selected for relatively low anthropogenic impact, the dominant sources of indicator bacteria were identified as various animals. The antibiotic resistance analysis technique promises to be a useful tool in assessing sources of fecal contamination in subtropical waters, such as those in Florida. PMID:10966379

  12. Decay of host-associated Bacteroidales cells and DNA in continuous-flow freshwater and seawater microcosms of identical experimental design and temperature as measured by PMA-qPCR and qPCR.

    PubMed

    Bae, Sungwoo; Wuertz, Stefan

    2015-03-01

    It is difficult to compare decay kinetics for genetic markers in an environmental context when they have been determined at different ambient temperatures. Therefore, we investigated the persistence of the host-associated genetic markers BacHum, BacCow and BacCan as well as the general Bacteroidales marker BacUni in both intact Bacteroidales cells and as total intracellular and extracellular marker DNA in controlled batch experiments at two temperatures using PMA-qPCR. Fecal Bacteroidales cells and DNA persisted longer at the lower temperature. Using the modified Arrhenius function to calculate decay constants for the same temperature, we then compared the decay of host-associated Bacteroidales cells and their DNA at 14 °C in field-based flow-through microcosms containing human, cow, and dog feces suspended in freshwater or seawater and previously operated with an identical experimental design. The time for a 2-log reduction (T₉₉) was used to characterize host-associated Bacteroidales decay. Host-associated genetic markers as determined by qPCR had similar T₉₉ values in freshwater and seawater at 14 °C when compared under both sunlight and dark conditions. In contrast, intact Bacteroidales cells measured by PMA-qPCR had shorter T₉₉ values in seawater than in freshwater. The decay constants of Bacteroidales cells were a function of physical (temperature) and chemical (salinity) parameters, suggesting that environmental parameters are key input variables for Bacteroidales survival in a predictive water quality model. Molecular markers targeting total Bacteroidales DNA were less susceptible to the variance of temperature, salinity and sunlight, implying that measurement of markers in both intact cells and DNA could enhance the predictive power of identifying fecal pollution across all aquatic environments. Monitoring Bacteroidales by qPCR alone rather than by PMA-qPCR does not always identify the contribution of recent fecal contamination because a

  13. Fecal-indicator bacteria in the Newfound Creek watershed, western North Carolina, during a high and low streamflow condition, 2003

    USGS Publications Warehouse

    Giddings, Elise M.; Oblinger, Carolyn J.

    2004-01-01

    Water quality in the Newfound Creek watershed has been shown to be affected by bacteria, sediment, and nutrients. In this study, Escherichia coli (E. coli) bacteria were sampled at five sites in Newfound Creek and five tributary sites during low flow on May 28, 2003, and high flow on November 19, 2003. In addition, a subset of five sites was sampled for fecal coliform bacteria, E. coli bacteria in streambed sediments (low flow only), and coliphage virus for serotyping. Coliphage virus serotyping has been used to identify human and animal sources of bacterial contamination. A streamflow gage was installed and operated to support ongoing water-quality studies in the watershed. Fecal coliform densities ranged from 92 to 27,000 colony-forming units per 100 milliliters of water for E. coli and 140 to an estimated 29,000 colony-forming units per 100 milliliters of water for fecal coliform during the two sampling visits. Ninety percent of the E. coli and fecal coliform samples exceeded corresponding U.S. Environmental Protection Agency or North Carolina water-quality criteria for recreational and ambient waters. During low flow, the middle part of the Newfound Creek watershed and the Dix Creek tributary had the highest densities of E. coli bacteria. During the high-flow sampling, all tributaries contained high densities of E. coli bacteria, although Dix Creek and Round Hill Branch were the largest contributors of these bacteria to Newfound Creek. Coliphage virus serotyping results were inconclusive because most samples did not contain the male-specific RNA coliphage needed for serotyping. Positive results indicated, however, that during low flow, non-human sources of bacteria were present in Sluder Branch, and during high flow, human sources of bacteria were present in Round Hill Branch. Sampling of bacteria in streambed sediments during low flow indicated that sediments do not appear to be a substantial source of bacteria relative to the water column, with the exception

  14. Detection of human-derived fecal contamination in Puerto Rico using carbamazepine, HF183 Bacteroides, and fecal indicator bacteria.

    PubMed

    Wade, Christina; Otero, Ernesto; Poon-Kwong, Brennan; Rozier, Ralph; Bachoon, Dave

    2015-12-30

    The level of fecal pollution in 17 sites in Puerto Rico was determined by Escherichia coli (E.coli) enumeration using an enzyme substrate medium and Quanti-Tray®/2000. Human fecal pollution was identified using an enzyme-linked immunosorbent assay for the detection of carbamazepine (CBZ) and quantitative polymerase chain reaction (qPCR) detection of the human Bacteroides marker, HF183. Carbamazepine was detected in 16 out of 17 sites, including Condado Lagoon, a popular recreational area. Elevated E.coli levels (>410 CFU 100 mL(-1)) were detected in 13 sites. Average CBZ concentrations ranged from 0.005 μg L(-1) to 0.482 μg L(-1) and 7 sites were positive for HF183. Higher CBZ concentrations were associated with the detection of HF183 (Mann-Whitney test; U=42.0; df=7; 1-tailed P value=0.013). This was the second study to determine surface water concentrations of CBZ in the Caribbean and the first in Puerto Rico. Copyright © 2015 Elsevier Ltd. All rights reserved.

  15. Fecal culture

    MedlinePlus

    ... fecal culture is a lab test to find organisms in the stool (feces) that can cause gastrointestinal ... Results There are no abnormal bacteria or other organisms in the sample. Talk to your provider about ...

  16. Efficacy of Vegetated Buffers in Preventing Transport of Fecal Coliform Bacteria from Pasturelands

    NASA Astrophysics Data System (ADS)

    Sullivan, Timothy J.; Moore, James A.; Thomas, David R.; Mallery, Eric; Snyder, Kai U.; Wustenberg, Mark; Wustenberg, Judith; Mackey, Sam D.; Moore, Deian L.

    2007-12-01

    An experimental study was conducted in Tillamook, Oregon, USA, to quantify the effectiveness of edge-of-field vegetated buffers for reducing transport of fecal coliform bacteria (FCB) from agricultural fields amended with dairy cow manure. Installation of vegetated buffers on loamy soils dramatically reduced the bacterial contamination of runoff water from manure-treated pasturelands, but the size of the vegetated buffer was not an important determinant of bacterial removal efficiency. Only 10% of the runoff samples collected from treatment cells having vegetated buffers exhibited FCB concentrations >200 colony forming units (cfu)/100 mL (a common water quality standard value), and the median concentration for all cells containing vegetated buffers was only 6 cfu/100 mL. The presence of a vegetated buffer of any size, from 1 to 25 m, generally reduced the median FCB concentration in runoff by more than 99%. Results for FCB load calculations were similar. Our results suggest that where substantial FCB contamination of runoff occurs from manure-treated pasturelands, it might be disproportionately associated with specific field or management conditions, such as the presence of soils that exhibit low water infiltration and generate larger volumes of runoff or the absence of a vegetated buffer. Buffer size regulations that do not consider such differences might not be efficient or effective in reducing bacterial contamination of runoff.

  17. Stratification and loading of fecal indicator bacteria (FIB) in a tidally muted urban salt marsh.

    PubMed

    Johnston, Karina K; Dorsey, John H; Saez, Jose A

    2015-03-01

    Stratification and loading of fecal indicator bacteria (FIB) were assessed in the main tidal channel of the Ballona Wetlands, an urban salt marsh receiving muted tidal flows, to (1) determine FIB concentration versus loading within the water column at differing tidal flows, (2) identify associations of FIB with other water quality parameters, and (3) compare wetland FIB concentrations to the adjacent estuary. Sampling was conducted four times during spring-tide events; samples were analyzed for FIB and turbidity (NTU) four times over a tidal cycle at pre-allocated depths, depending on the water level. Additional water quality parameters measured included temperature, salinity, oxygen, and pH. Loadings were calculated by integrating the stratified FIB concentrations with water column cross-sectional volumes corresponding to each depth. Enterococci and Escherichia coli were stratified both by concentration and loading, although these variables portrayed different patterns over a tidal cycle. Greatest concentrations occurred in surface to mid-strata levels, during flood tides when contaminated water flowed in from the estuary, and during ebb flows when sediments were suspended. Loading was greatest during flood flows and diminished during low tide periods. FIB concentrations within the estuary often were significantly greater than those within the wetland tide channel, supporting previous studies that the wetlands act as a sink for FIB. For public health water quality monitoring, these results indicate that more accurate estimates of FIB concentrations would be obtained by sampling a number of points within a water column rather than relying only on single surface samples.

  18. Predictive Power of Clean Bed Filtration Theory for Fecal Indicator Bacteria Removal in Stormwater Biofilters.

    PubMed

    Parker, Emily A; Rippy, Megan A; Mehring, Andrew S; Winfrey, Brandon K; Ambrose, Richard F; Levin, Lisa A; Grant, Stanley B

    2017-05-16

    Green infrastructure (also referred to as low impact development, or LID) has the potential to transform urban stormwater runoff from an environmental threat to a valuable water resource. In this paper we focus on the removal of fecal indicator bacteria (FIB, a pollutant responsible for runoff-associated inland and coastal beach closures) in stormwater biofilters (a common type of green infrastructure). Drawing on a combination of previously published and new laboratory studies of FIB removal in biofilters, we find that 66% of the variance in FIB removal rates can be explained by clean bed filtration theory (CBFT, 31%), antecedent dry period (14%), study effect (8%), biofilter age (7%), and the presence or absence of shrubs (6%). Our analysis suggests that, with the exception of shrubs, plants affect FIB removal indirectly by changing the infiltration rate, not directly by changing the FIB removal mechanisms or altering filtration rates in ways not already accounted for by CBFT. The analysis presented here represents a significant step forward in our understanding of how physicochemical theories (such as CBFT) can be melded with hydrology, engineering design, and ecology to improve the water quality benefits of green infrastructure.

  19. Groundwater discharge: potential association with fecal indicator bacteria in the surf zone.

    PubMed

    Boehm, Alexandria B; Shellenbarger, Gregory G; Paytan, Adina

    2004-07-01

    Short-lived radium isotopes (223Ra and 224Ra) are used to investigate the potential association between groundwater discharge and microbial pollution at Huntington Beach, CA. We establish the tidally driven exchange of groundwater from the surficial beach aquifer across the beach face. Groundwater is found to be a source of nutrients (silica, inorganic nitrogen, and orthophosphate) to the surf zone, and these nutrients could possibly provide an environment for enhanced growth or increased persistence of fecal indicator bacteria (FIB). Ammonium and ortho-phosphate explain up to 12-20% of the variance in FIB levels in the surf zone. Elevated levels of FIB were only found in 1 of the 26 groundwater samples. However, FIB in the surf zone covary with radium at fortnightly, diurnal, and semi-diurnal tidal periods. In addition, radium accounts for up to 38% of the variance in log-FIB levels in the surf zone. A column experiment illustrates that Enterococcus suspended in Huntington Beach saline groundwater is not significantly filtered by sand collected from the field. This work establishes a mechanism for the subterranean delivery of FIB pollution to the surf zone from the surficial aquifer and presents evidence that supports an association between groundwater discharge and FIB.

  20. Detection of Helicobacter pylori and fecal indicator bacteria in five North American rivers.

    USGS Publications Warehouse

    Voytek, M.A.; Ashen, J.B.; Fogarty, L.R.; Kirshtein, J.D.; Landa, E.R.

    2005-01-01

    This study examines the use of fecal indicator bacteria (FIB) as a predictor of the presence of Helicobacter spp. A combination of standard culture and molecular techniques were used to detect and quantify FIB, Helicobacter spp. and H. pylori from five North American rivers of different size and with different land use characteristics. Primers designed to amplify genes specific to Helicobacter spp. and H. pylori were evaluated for their efficacy in detection and quantification in environmental samples. Helicobacter spp. were detected in 18/33 (55%) of river samples. H. pylori was detected in 11/33 (33%) of river samples. FIB were found in 32/33 (96%) of river samples. When FIB abundance exceeded USEPA water quality standards for single samples, Helicobacter or H. pylori were detected in 7/15 (47%) cases. No numerical correlation was found between the presence of FIB and either Helicobacter spp. or H. pylori. This suggests that the presence of FIB will be of limited use for detection of Helicobacter spp. or H. pylori by public health agencies.

  1. Treating Stormwater with Green Infrastructure: Plants, Residence Time Distributions, and the Removal of Fecal Indicator Bacteria

    NASA Astrophysics Data System (ADS)

    Parker, E.; Grant, S. B.; Rippy, M.; Winfrey, B.; Mehring, A.

    2015-12-01

    In many cities, green infrastructure is increasingly used to capture and treat stormwater runoff, due to the many opportunities these systems afford for protecting receiving water quality and ecology while mitigating water scarcity. Here, we focus on how plants affect the removal of fecal indicator bacteria (FIB) in newly-constructed stormwater biofilters, a type of green infrastructure consisting of unconsolidated granular media containing one or more plant species. Input-response experiments were carried out using both non-reactive (salt) and reactive (sewage) tracers on six laboratory-scale (~1m long by 24 cm diameter) biofilters, half of which were planted with the sedge Carex appressa (treatment replicates) and half of which were unplanted (control replicates). C. appressa modifies the residence time distribution (RTD) in a biofilter by creating preferential flow paths along which water and mass can move quickly, but does not appear to alter the intrinsic rate at which FIB are removed. Thus, the "green" component of green infrastructure can alter pollutant removal by changing the RTD, with or without a concomitant change in pollutant reactivity.

  2. Associations between fecal indicator bacteria prevalence and demographic data in private water supplies in Virginia.

    PubMed

    Smith, Tamara; Krometis, Leigh-Anne H; Hagedorn, Charles; Lawrence, Annie H; Benham, Brian; Ling, Erin; Ziegler, Peter; Marmagas, Susan West

    2014-12-01

    Over 1.7 million Virginians rely on private water sources to provide household water. The heaviest reliance on these systems occurs in rural areas, which are often underserved with respect to available financial resources and access to environmental health education. This study aimed to identify potential associations between concentrations of fecal indicator bacteria (FIB) (coliforms, Escherichia coli) in over 800 samples collected at the point-of-use from homes with private water supply systems and homeowner-provided demographic data (household income and education). Of the 828 samples tested, 349 (42%) of samples tested positive for total coliform and 55 (6.6%) tested positive for E. coli. Source tracking efforts targeting optical brightener concentrations via fluorometry and the presence of a human-specific Bacteroides marker via quantitative real-time polymerase chain reaction (qPCR) suggest possible contamination from human septage in over 20 samples. Statistical methods implied that household income has an association with the proportion of samples positive for total coliform, though the relationship between education level and FIB is less clear. Further exploration of links between demographic data and private water quality will be helpful in building effective strategies to improve rural drinking water quality.

  3. Spatio-Temporal Variability in Fecal Indicator Bacteria Concentrations at Huntington Beach: Connections to Physical Forcing

    NASA Astrophysics Data System (ADS)

    Rippy, M. A.; Feddersen, F.; Leichter, J.; Omand, M.; Moore, D. F.; McGee, C.; Franks, P. J.

    2007-05-01

    Two major factors determine the spatial and temporal distributions of fecal indicator bacteria (FIB) at a given beach: local circulation & mixing patterns, and bacterial inactivation rates. High frequency and spatial resolution bacterial sampling combined with measurements of physical processes can be used to infer inactivation rates, enabling differentiation between dilution & mortality as factors driving variability in nearshore FIB abundance. A FIB sampling experiment (HB06) took place on 16 October 2006, at Huntington State Beach, a site selected due to its persistent problems with FIB pollution. Water samples were taken at 20-minute intervals (from 6:50am to 11:50am) at ten locations; four in an alongshore transect spanning 1 km at the shoreline, and the remainder in a 300-m long cross-shore transect. All samples were analyzed for FIB concentration (Total Coliforms, E. coli & Enterococci) and, for a subset, species level Enterococcus composition was determined. As part of the HB06 experiment, currents, temperature, waves, and chlorophyll fluorescence were measured simultaneously in the cross-shore direction with rapid CTD casts 300 m offshore. Results indicate that E. coli and Enterococcus concentrations exhibit exponential decreases with time, with smaller decay rates associated with depth and with sites in the Talbert Marsh and Santa Ana River. FIB concentrations are also noticeably lower farther offshore (300 m). Spatio-temporal patterns in FIB concentration will be presented in conjunction with the nearshore physical data allowing the relationship between physical dynamics and biological variability to be addressed.

  4. Quantification of human norovirus GII, human adenovirus, and fecal indicator organisms in wastewater used for irrigation in Accra, Ghana.

    PubMed

    Silverman, Andrea I; Akrong, Mark O; Amoah, Philip; Drechsel, Pay; Nelson, Kara L

    2013-09-01

    Quantitative microbial risk assessment (QMRA) is frequently used to estimate health risks associated with wastewater irrigation and requires pathogen concentration estimates as inputs. However, human pathogens, such as viruses, are rarely quantified in water samples, and simple relationships between fecal indicator bacteria and pathogen concentrations are used instead. To provide data that can be used to refine QMRA models of wastewater-fed agriculture in Accra, stream, drain, and waste stabilization pond waters used for irrigation were sampled and analyzed for concentrations of fecal indicator microorganisms (human-specific Bacteroidales, Escherichia coli, enterococci, thermotolerant coliform, and somatic and F+ coliphages) and two human viruses (adenovirus and norovirus genogroup II). E. coli concentrations in all samples exceeded limits suggested by the World Health Organization, and human-specific Bacteroidales was found in all but one sample, suggesting human fecal contamination. Human viruses were detected in 16 out of 20 samples, were quantified in 12, and contained 2-3 orders of magnitude more norovirus than predicted by norovirus to E. coli concentration ratios assumed in recent publications employing indicator-based QMRA. As wastewater irrigation can be beneficial for farmers and municipalities, these results should not discourage water reuse in agriculture, but provide motivation and targets for wastewater treatment before use on farms.

  5. Modeling fecal bacteria transport and retention in agricultural and urban soils under saturated and unsaturated flow conditions.

    PubMed

    Balkhair, Khaled S

    2017-03-01

    Pathogenic bacteria, that enter surface water bodies and groundwater systems through unmanaged wastewater land application, pose a great risk to human health. In this study, six soil column experiments were conducted to simulate the vulnerability of agricultural and urban field soils for fecal bacteria transport and retention under saturated and unsaturated flow conditions. HYDRUS-1D kinetic attachment and kinetic attachment-detachment models were used to simulate the breakthrough curves of the experimental data by fitting model parameters. Results indicated significant differences in the retention and drainage of bacteria between saturated and unsaturated flow condition in the two studied soils. Flow under unsaturated condition retained more bacteria than the saturated flow case. The high bacteria retention in the urban soil compared to agricultural soil is ascribed not only to the dynamic attachment and sorption mechanisms but also to the greater surface area of fine particles and low flow rate. All models simulated experimental data satisfactorily under saturated flow conditions; however, under variably saturated flow, the peak concentrations were overestimated by the attachment-detachment model and underestimated by the attachment model with blocking. The good match between observed data and simulated concentrations by the attachment model which was supported by the Akaike information criterion (AIC) for model selection indicates that the first-order attachment coefficient was sufficient to represent the quantitative and temporal distribution of bacteria in the soil column. On the other hand, the total mass balance of the drained and retained bacteria in all transport experiments was in the range of values commonly found in the literature. Regardless of flow conditions and soil texture, most of the bacteria were retained in the top 12 cm of the soil column. The approaches and the models used in this study have proven to be a good tool for simulating fecal

  6. Molecular detection of Campylobacter spp. and fecal indicator bacteria during the northern migration of Sandhill Cranes (Grus canadensis) at the Central Platte River

    EPA Science Inventory

    The annual Sandhill crane (Grus canadensis) migration through Nebraska is thought to be a major source of fecal pollution to the Platte River, but of unknown human health risk. To better understand potential risks, the presence of Campylobacter species and fecal bacteria were exa...

  7. Molecular detection of Campylobacter spp. and fecal indicator bacteria during the northern migration of Sandhill Cranes (Grus canadensis) at the Central Platte River

    EPA Science Inventory

    The annual Sandhill crane (Grus canadensis) migration through Nebraska is thought to be a major source of fecal pollution to the Platte River, but of unknown human health risk. To better understand potential risks, the presence of Campylobacter species and fecal bacteria were exa...

  8. Salmonella and fecal indicator bacteria in tile waters draining poultry litter application fields in central Iowa

    NASA Astrophysics Data System (ADS)

    Hruby, C.; Soupir, M.

    2012-12-01

    E. coli and enterococci are commonly used as pathogen indicators in surface waters. Along with these indicators, pathogenic Salmonella are prevalent in poultry litter, and have the potential to be transported from land-application areas to tile waters and ultimately to impact waters that are used for drinking-water and recreation. The fate and transport of these bacteria to drainage tiles from application fields, and the correlation of fecal indicator bacteria to pathogens in this setting, is poorly understood. In this field study, samples were obtained from poultry litter, soil, and drainage tile waters below chisel-plowed and no-till cornfields in central Iowa where poultry litter was applied each year in late spring prior to planting. Litter was applied at three different rates; commercial fertilizer with no litter, a low application rate based on the nitrogen requirements of the corn (PL1), and double the low rate (PL2). This site is characterized by low sloping (0-9%) Clarion and Nicollet soils, which are derived from glacial till. Samples were collected from April to September for three years (2010-12) when tiles were flowing. Record high precipitation fell during the sampling period in 2010, while 2011 and 2012 were exceptionally dry years at this location. Grab samples were taken directly from flowing tiles after every rainfall event (>2 cm in less than 24 hours) and samples were collected hourly throughout selected events using an automatic sampling device. Concentrations of E. coli, enterococci and Salmonella spp. were quantified by membrane filtration and growth on selective agars. Peak bacteria concentrations following rainfall events were often one order of magnitude higher in tile waters discharging from no-till plots, despite the smaller size and lower tile flow rates at these plots compared to the chisel-plowed plots. Bacteria concentrations regularly varied by two orders of magnitude in response to rainfall events. Bacteria transport via macropores

  9. Comparison and continuous estimates of fecal coliform and Escherichia coli bacteria in selected Kansas streams, May 1999 through April 2002

    USGS Publications Warehouse

    Rasmussen, Patrick P.; Ziegler, Andrew C.

    2003-01-01

    The sanitary quality of water and its use as a public-water supply and for recreational activities, such as swimming, wading, boating, and fishing, can be evaluated on the basis of fecal coliform and Escherichia coli (E. coli) bacteria densities. This report describes the overall sanitary quality of surface water in selected Kansas streams, the relation between fecal coliform and E. coli, the relation between turbidity and bacteria densities, and how continuous bacteria estimates can be used to evaluate the water-quality conditions in selected Kansas streams. Samples for fecal coliform and E. coli were collected at 28 surface-water sites in Kansas. Of the 318 samples collected, 18 percent exceeded the current Kansas Department of Health and Environment (KDHE) secondary contact recreational, single-sample criterion for fecal coliform (2,000 colonies per 100 milliliters of water). Of the 219 samples collected during the recreation months (April 1 through October 31), 21 percent exceeded the current (2003) KDHE single-sample fecal coliform criterion for secondary contact rec-reation (2,000 colonies per 100 milliliters of water) and 36 percent exceeded the U.S. Environmental Protection Agency (USEPA) recommended single-sample primary contact recreational criterion for E. coli (576 colonies per 100 milliliters of water). Comparisons of fecal coliform and E. coli criteria indicated that more than one-half of the streams sampled could exceed USEPA recommended E. coli criteria more frequently than the current KDHE fecal coliform criteria. In addition, the ratios of E. coli to fecal coliform (EC/FC) were smallest for sites with slightly saline water (specific conductance greater than 1,000 microsiemens per centimeter at 25 degrees Celsius), indicating that E. coli may not be a good indicator of sanitary quality for those streams. Enterococci bacteria may provide a more accurate assessment of the potential for swimming-related illnesses in these streams. Ratios of EC/FC and

  10. Validation of the H2S method to detect bacteria of fecal origin by cultured and molecular methods.

    PubMed

    McMahan, Lanakila; Devine, Anthony A; Grunden, Amy M; Sobsey, Mark D

    2011-12-01

    Using biochemical and molecular methods, this research determined whether or not the H(2)S test did correctly identify sewage-contaminated waters by being the first to use culturing and molecular methods to identify the types and numbers of fecal indicator organisms, pathogens, and other microbes present in sewage samples with positive H(2)S test results. For the culture-based method, samples were analyzed for the presence of fecal bacteria by spread plating the sewage sample onto differential and selective media for Aeromonas spp., Escherichia coli, sulfite-reducing clostridia, H(2)S-producing bacteria, and Salmonella/Shigella spp. The isolates were then: (1) tested to determine whether they were H(2)S-producing organisms and (2) identified to the genus and species level using biochemical methods. The molecular method used to characterize the microbial populations of select samples was terminal restriction fragment length polymorphisms. These experiments on sewage provided evidence that positive H(2)S tests consistently contained fecal bacteria and pathogens. There were strong relationships of agreement between the organisms identified by both methods tested. This study is an important advance in microbial water quality detection since it is focused on the evaluation of a novel, low-cost, water microbiology test that has the potential to provide millions of people worldwide access to water quality detection technology. Of prime consideration in evaluating water quality tests is the determination of the test's accuracy and specificity, and this article is a fundamental step in providing that information.

  11. Survival of Salmonella spp. and fecal indicator bacteria in Vietnamese biogas digesters receiving pig slurry.

    PubMed

    Huong, Luu Quynh; Forslund, Anita; Madsen, Henry; Dalsgaard, Anders

    2014-09-01

    Small-scale biogas digesters are widely promoted worldwide as a sustainable technology to manage livestock manure. In Vietnam, pig slurry is commonly applied to biogas digesters for production of gas for electricity and cooking with the effluent being used to fertilize field crops, vegetables and fish ponds. Slurry may contain a variety of zoonotic pathogens, e.g. Salmonella spp., which are able to cause disease in humans either through direct contact with slurry or by fecal contamination of water and foods. The objective of this study was to evaluate the survival of Salmonella spp. and the fecal indicator bacteria, enterococci, E. coli, and spores of Clostridium perfringens in biogas digesters operated by small-scale Vietnamese pig farmers. The serovar and antimicrobial susceptibility of the Salmonella spp. isolated were also established. The study was conducted in 12 farms (6 farms with and 6 farms without toilet connected) located in Hanam province, Vietnam. Sampling of pig slurry and biogas effluent was done during two seasons. Results showed that the concentration of enterococci, E. coli, and Clostridium perfringens spores was overall reduced by only 1-2 log10-units in the biogas digesters when comparing raw slurry and biogas effluent. Salmonella spp. was found in both raw slurry and biogas effluent. A total of 19 Salmonella serovars were identified, with the main serovars being Salmonella Typhimurium (55/138), Salmonella enterica serovar 4,[5],12:i:- (19/138), Salmonella Weltevreden (9/138) and Salmonella Rissen (9/138). The Salmonella serovars showed similar antimicrobial resistance patterns to those previously reported from Vietnam. When promoting biogas, farmers should be made aware that effluent should only be used as fertilizer for crops not consumed raw and that indiscriminate discharge of effluent are likely to contaminate water recipients, e.g. drinking water sources, with pathogens. Relevant authorities should promote safe animal manure management

  12. Biotic interactions and sunlight affect persistence of fecal indicator bacteria and microbial source tracking genetic markers in the Upper Mississippi River

    EPA Science Inventory

    Sanitary quality of recreational waters is assessed by enumerating fecal indicator bacteria (FIB) (Escherichia coli and enterococci); organisms present in the gastrointestinal tract of humans and many other animals, hence providing no information about the pollution source. Micro...

  13. Biotic interactions and sunlight affect persistence of fecal indicator bacteria and microbial source tracking genetic markers in the Upper Mississippi River

    EPA Science Inventory

    Sanitary quality of recreational waters is assessed by enumerating fecal indicator bacteria (FIB) (Escherichia coli and enterococci); organisms present in the gastrointestinal tract of humans and many other animals, hence providing no information about the pollution source. Micro...

  14. Community structures of fecal bacteria in cattle from different animal feeding operations

    EPA Science Inventory

    The fecal microbiome of cattle plays a critical role not only in animal health and productivity, but also in methane emissions, food safety, pathogen shedding, and the performance of fecal pollution detection methods. Unfortunately, most published molecular surveys fail to provid...

  15. Community structures of fecal bacteria in cattle from different animal feeding operations

    EPA Science Inventory

    The fecal microbiome of cattle plays a critical role not only in animal health and productivity, but also in methane emissions, food safety, pathogen shedding, and the performance of fecal pollution detection methods. Unfortunately, most published molecular surveys fail to provid...

  16. Community structure of cattle fecal bacteria from different animal feeding operations

    USDA-ARS?s Scientific Manuscript database

    The fecal microbiome of cattle plays a critical role not only in animal health and productivity, but also in methane emissions, food safety, pathogen shedding, and the performance of fecal pollution detection methods. Unfortunately, most published molecular surveys fail to provide adequate detail ab...

  17. Fecal indicator bacteria and Salmonella in ponds managed as bird habitat, San Francisco Bay, California, USA.

    PubMed

    Shellenbarger, Gregory G; Athearn, Nicole D; Takekawa, John Y; Boehm, Alexandria B

    2008-06-01

    Throughout the world, coastal resource managers are encouraging the restoration of previously modified coastal habitats back into wetlands and managed ponds for their ecosystem value. Because many coastal wetlands are adjacent to urban centers and waters used for human recreation, it is important to understand how wildlife can affect water quality. We measured fecal indicator bacteria (FIB) concentrations, presence/absence of Salmonella, bird abundance, and physico-chemical parameters in two coastal, managed ponds and adjacent sloughs for 4 weeks during the summer and winter in 2006. We characterized the microbial water quality in these waters relative to state water-quality standards and examined the relationship between FIB, bird abundance, and physico-chemical parameters. A box model approach was utilized to determine the net source or sink of FIB in the ponds during the study periods. FIB concentrations often exceeded state standards, particularly in the summer, and microbial water quality in the sloughs was generally lower than in ponds during both seasons. Specifically, the inflow of water from the sloughs to the ponds during the summer, more so than waterfowl use, appeared to increase the FIB concentrations in the ponds. The box model results suggested that the ponds served as net wetland sources and sinks for FIB, and high bird abundances in the winter likely contributed to net winter source terms for two of the three FIB in both ponds. Eight serovars of the human pathogen Salmonella were isolated from slough and pond waters, although the source of the pathogen to these wetlands was not identified. Thus, it appeared that factors other than bird abundance were most important in modulating FIB concentrations in these ponds.

  18. Fecal indicator bacteria and Salmonella in ponds managed as bird habitat, San Francisco Bay, California, USA

    USGS Publications Warehouse

    Shellenbarger, G.G.; Athearn, N.D.; Takekawa, John Y.; Boehm, A.B.

    2008-01-01

    Throughout the world, coastal resource managers are encouraging the restoration of previously modified coastal habitats back into wetlands and managed ponds for their ecosystem value. Because many coastal wetlands are adjacent to urban centers and waters used for human recreation, it is important to understand how wildlife can affect water quality. We measured fecal indicator bacteria (FIB) concentrations, presence/absence of Salmonella, bird abundance, and physico-chemical parameters in two coastal, managed ponds and adjacent sloughs for 4 weeks during the summer and winter in 2006. We characterized the microbial water quality in these waters relative to state water-quality standards and examined the relationship between FIB, bird abundance, and physico-chemical parameters. A box model approach was utilized to determine the net source or sink of FIB in the ponds during the study periods. FIB concentrations often exceeded state standards, particularly in the summer, and microbial water quality in the sloughs was generally lower than in ponds during both seasons. Specifically, the inflow of water from the sloughs to the ponds during the summer, more so than waterfowl use, appeared to increase the FIB concentrations in the ponds. The box model results suggested that the ponds served as net wetland sources and sinks for FIB, and high bird abundances in the winter likely contributed to net winter source terms for two of the three FIB in both ponds. Eight serovars of the human pathogen Salmonella were isolated from slough and pond waters, although the source of the pathogen to these wetlands was not identified. Thus, it appeared that factors other than bird abundance were most important in modulating FIB concentrations in these ponds.

  19. Holding-time and method comparisons for the analysis of fecal-indicator bacteria in groundwater.

    PubMed

    Bushon, Rebecca N; Brady, Amie M G; Lindsey, Bruce D

    2015-11-01

    As part of the US Geological Survey National Water-Quality Assessment Program, groundwater samples from domestic- and public-supply wells were collected and analyzed for fecal-indicator bacteria. A holding time comparison for total coliforms, Escherichia coli, and enterococci was done by analyzing samples within 8 h using presence/absence methods and within 18-30 h using quantitative methods. The data indicate that results obtained within 18-30 h were not significantly different from those obtained within 8 h for total coliforms and enterococci, by Colilert® and Enterolert® methods (IDEXX Laboratories Inc., Westbrook, ME), respectively. Quantitative laboratory methods for samples analyzed within 18-30 h showed a statistically significant higher detection frequency when compared to presence/absence methods done within 8 h for the following methods, E. coli by Colilert and enterococci by membrane filtration on mEI agar. Additionally, a comparison of methods for the enumeration of enterococci was done. Using non-parametric statistical analyses, results from the two methods were statistically different. In this study, the membrane filtration method on mEI agar was more sensitive, resulted in more detections of enterococci, and results were easier to interpret than with the quantitative Enterolert method. The quantitative Enterolert method produced varying levels of fluorescence, which required additional verification steps to eliminate false-positive results. It may be more advantageous to analyze untreated groundwater for enterococci using the membrane filtration method on mEI agar.

  20. Narrowing the Search for Sources of Fecal Indicator Bacteria with a Simple Salinity Mixing Model

    NASA Astrophysics Data System (ADS)

    McLaughlin, K.; Ahn, J.; Litton, R.; Grant, S. B.

    2006-12-01

    Newport Bay, the second largest estuarine embayment in Southern California, provides critical natural habitat for terrestrial and aquatic species and is a regionally important recreational area. Unfortunately, the beneficial uses of Newport Bay are threatened by numerous sources of pollutant loading, either through direct discharge into the bay or through its tributaries. Fecal indicator bacteria (FIB) are associated with human pathogens and are present in high concentrations in sewage and urban runoff. Standardized and inexpensive assays used for the detection of FIB have allowed their concentrations to be used as a common test of water quality. In order to assess FIB impairment in Newport Bay, weekly transects of FIB concentrations were conducted, specifically Total Coliform, Escherichia coli and Enterococci spp., as well as salinity, temperature, and transmissivity, from the upper reaches of the estuary to an offshore control site. Using salinity as a conservative tracer for water mass mixing and determining the end-member values of FIB and transmissivity in both the creek sites and the offshore control site, we created a simple, two end-member mixing model of FIB and transmissivity within Newport Bay. Deviations from the mixing model would suggest either an additional source of FIB to the bay (e.g. bird feces) or regrowth of FIB within the bay. Our results indicate that, with a few notable exceptions, salinity is a good tracer for FIB concentrations along the transect, but is not particularly effective for transmissivity. This suggests that the largest contributor of FIB loading to Newport Bay comes from the discharge of creeks into the upper reaches of the estuary.

  1. Locating sources of surf zone pollution: a mass budget analysis of fecal indicator bacteria at Huntington Beach, California.

    PubMed

    Kim, Joon Ha; Grant, Stanley B; McGee, Charles D; Sanders, Brett F; Largier, John L

    2004-05-01

    The surf zone is the unique environment where ocean meets land and a place of critical ecological, economic, and recreational importance. In the United States, this natural resource is increasingly off-limits to the public due to elevated concentrations of fecal indicator bacteria and other contaminants, the sources of which are often unknown. In this paper, we describe an approach for calculating mass budgets of pollutants in the surf zone from shoreline monitoring data. The analysis reveals that fecal indicator bacteria pollution in the surf zone at several contiguous beaches in Orange County, California, originates from well-defined locations along the shore, including the tidal outlets of the Santa Ana River and Talbert Marsh. Fecal pollution flows into the ocean from the Santa Ana River and Talbert Marsh outlets during ebb tides and from there is transported parallel to the shoreline by wave-driven surf zone currents and/or offshore tidal currents, frequently contaminating >5 km of the surf zone. The methodology developed here for locating and quantifying sources of surf zone pollution should be applicable to a wide array of contaminants and coastal settings.

  2. Microbial source tracking of private well water samples across at-risk regions in southern Ontario and analysis of traditional fecal indicator bacteria assays including culture and qPCR.

    PubMed

    Krolik, Julia; Maier, Allison; Thompson, Shawna; Majury, Anna

    2016-12-01

    Many people living in rural areas rely on privately owned wells as their primary source of drinking water. These water sources are at risk for fecal contamination of human, wildlife, and livestock origin. While traditional bacteriological testing involves culture-based methods, microbial source tracking (MST) assays present an opportunity to additionally determine the source of fecal contamination. This study investigated the main host sources of contamination in private well water samples with high levels of Escherichia coli (E. coli), using MST with human and multi-species specific markers. Fecal contamination of human origin was detected in approximately 50% of samples, indicating that current contamination prevention strategies require reconsideration. The relationship between cattle density and fecal contamination of bovine origin was investigated using a Bovine Bacteroidales specific MST assay. Regional variations of microbial sources were examined, and may inform local primary prevention strategies. Additionally, in order to assess MST and E. coli quantitative real time polymerase chain reaction (qPCR) assays as indicators of fecal contamination, these were compared to E. coli culture methods. Variation in results was observed across all assay methods investigated, suggesting the most appropriate routine bacteriological testing methodology cannot be determined without comparison to a method that directly detects the presence of fecal contamination.

  3. Microbial community analysis and identification of alternative host-specific fecal indicators in fecal and river water samples using pyrosequencing.

    PubMed

    Jeong, Ju-Yong; Park, Hee-Deung; Lee, Kyong-Hee; Weon, Hang-Yeon; Ka, Jong-Ok

    2011-08-01

    It is important to know the comprehensive microbial communities of fecal pollution sources and receiving water bodies for microbial source tracking. Pyrosequencing targeting the V1-V3 hypervariable regions of the 16S rRNA gene was used to investigate the characteristics of bacterial and Bacteroidales communities in major fecal sources and river waters. Diversity analysis indicated that cow feces had the highest diversities in the bacterial and Bacteroidales group followed by the pig sample, with human feces having the lowest value. The Bacteroidales, one of the potential fecal indicators, totally dominated in the fecal samples accounting for 31%-52% of bacterial sequences, but much less (0.6%) in the river water. Clustering and Venn diagram analyses showed that the human sample had a greater similarity to the pig sample in the bacterial and Bacteroidales communities than to samples from other hosts. Traditional fecal indicators, i.e., Escherichia coli, were detected in the human and river water samples at very low rates and Clostridium perfringens and enterococci were not detected in any samples. Besides the Bacteroidales group, some microorganisms detected in the specific hosts, i.e., Parasutterella excrementihominis, Veillonella sp., Dialister invisus, Megamonas funiformis, and Ruminococcus lactaris for the human and Lactobacillus amylovorus and Atopostipes sp. for the pig, could be used as potential host-specific fecal indicators. These microorganisms could be used as multiple fecal indicators that are not dependent on the absence or presence of a single indicator. Monitoring for multiple indicators that are highly abundant and host-specific would greatly enhance the effectiveness of fecal pollution source tracking.

  4. Antibiotic-resistant fecal bacteria, antibiotics, and mercury in surface waters of Oakland County, Michigan, 2005-2006

    USGS Publications Warehouse

    Fogarty, Lisa R.; Duris, Joseph W.; Crowley, Suzanne L.; Hardigan, Nicole

    2007-01-01

    Water samples collected from 20 stream sites in Oakland and Macomb Counties, Mich., were analyzed to learn more about the occurrence of cephalosporin-resistant Escherichia coli (E. coli) and vancomycin-resistant enterococci (VRE) and the co-occurrence of antibiotics and mercury in area streams. Fecal indicator bacteria concentrations exceeded the Michigan recreational water-quality standard of 300 E. coli colony forming units (CFU) per 100 milliliters of water in 19 of 35 stream-water samples collected in Oakland County. A gene commonly associated with enterococci from humans was detected in samples from Paint Creek at Rochester and Evans Ditch at Southfield, indicating that human fecal waste is a possible source of fecal contamination at these sites. E. coli resistant to the cephalosporin antibiotics (cefoxitin and/ or ceftriaxone) were found at all sites on at least one occasion. The highest percentages of E. coli isolates resistant to cefoxitin and ceftriaxone were 71 percent (Clinton River at Auburn Hills) and 19 percent (Sashabaw Creek near Drayton Plains), respectively. Cephalosporin-resistant E. coli was detected more frequently in samples from intensively urbanized or industrialized areas than in samples from less urbanized areas. VRE were not detected in any sample collected in this study. Multiple antibiotics (azithromycin, erythromycin, ofloxacin, sulfamethoxazole, and trimethoprim) were detected in water samples from the Clinton River at Auburn Hills, and tylosin (an antibiotic used in veterinary medicine and livestock production that belongs to the macrolide group, along with erythromycin) was detected in one water sample from Paint Creek at Rochester. Concentrations of total mercury were as high as 19.8 nanograms per liter (Evans Ditch at Southfield). There was no relation among percentage of antibiotic-resistant bacteria and measured concentrations of antibiotics or mercury in the water. Genetic elements capable of exchanging multiple antibiotic

  5. Fecal coliform and Escherichia coli bacteria in the St. Croix National Scenic Riverway, Summer 1999

    USGS Publications Warehouse

    Kroening, Sharon E.

    1999-01-01

    Fecal coliform and Escherichia coli (E. coli) concentrations were determined in the St. Croix National Scenic Riverway to assess whether pathogenic organisms pose a potential problem for recreational use. Samples were collected from May through September 1999 at 22 locations on the St. Croix and Namekagon Rivers. No concentrations exceeded water-quality criteria or standards set by the U.S. Environmental Protection Agency or the states of Minnesota and Wisconsin. Maximum fecal coliform and E. coli concentrations were measured in the St. Croix River at St. Croix Falls, Wisconsin. Median fecal coliform and E. coli concentrations were greater in the St. Croix River near Woodland Corner, Wisconsin, and at Marine on St. Croix, Minnesota than at other locations sampled. There were no consistent short-term variations in fecal coliform or E. coli concentrations during the summer period or any significant relations between concentrations and stream discharge, based on these results.

  6. Source tracking swine fecal waste in surface water proximal to swine concentrated animal feeding operations

    PubMed Central

    Heaney, Christopher D.; Myers, Kevin; Wing, Steve; Hall, Devon; Baron, Dothula; Stewart, Jill R.

    2015-01-01

    Swine farming has gone through many changes in the last few decades, resulting in operations with a high animal density known as confined animal feeding operations (CAFOs). These operations produce a large quantity of fecal waste whose environmental impacts are not well understood. The purpose of this study was to investigate microbial water quality in surface waters proximal to swine CAFOs including microbial source tracking of fecal microbes specific to swine. For one year, surface water samples at up- and downstream sites proximal to swine CAFO lagoon waste land application sites were tested for fecal indicator bacteria (fecal coliforms, Escherichia coli and Enterococcus) and candidate swine-specific microbial source-tracking (MST) markers (Bacteroidales Pig-1-Bac, Pig-2-Bac, and Pig-Bac-2, and methanogen P23-2). Testing of 187 samples showed high fecal indicator bacteria concentrations at both up- and downstream sites. Overall, 40%, 23%, and 61% of samples exceeded state and federal recreational water quality guidelines for fecal coliforms, E. coli, and Enterococcus, respectively. Pig-1-Bac and Pig-2-Bac showed the highest specificity to swine fecal wastes and were 2.47 (95% confidence interval [CI] = 1.03, 5.94) and 2.30 times (95% CI = 0.90, 5.88) as prevalent proximal down- than proximal upstream of swine CAFOs, respectively. Pig-1-Bac and Pig-2-Bac were also 2.87 (95% CI = 1.21, 6.80) and 3.36 (95% CI = 1.34, 8.41) times as prevalent when 48 hour antecedent rainfall was greater than versus less than the mean, respectively. Results suggest diffuse and overall poor sanitary quality of surface waters where swine CAFO density is high. Pig-1-Bac and Pig-2-Bac are useful for tracking off-site conveyance of swine fecal wastes into surface waters proximal to and downstream of swine CAFOs and during rain events. PMID:25600418

  7. Source tracking swine fecal waste in surface water proximal to swine concentrated animal feeding operations.

    PubMed

    Heaney, Christopher D; Myers, Kevin; Wing, Steve; Hall, Devon; Baron, Dothula; Stewart, Jill R

    2015-04-01

    Swine farming has gone through many changes in the last few decades, resulting in operations with a high animal density known as confined animal feeding operations (CAFOs). These operations produce a large quantity of fecal waste whose environmental impacts are not well understood. The purpose of this study was to investigate microbial water quality in surface waters proximal to swine CAFOs including microbial source tracking of fecal microbes specific to swine. For one year, surface water samples at up- and downstream sites proximal to swine CAFO lagoon waste land application sites were tested for fecal indicator bacteria (fecal coliforms, Escherichia coli and Enterococcus) and candidate swine-specific microbial source-tracking (MST) markers (Bacteroidales Pig-1-Bac, Pig-2-Bac, and Pig-Bac-2, and methanogen P23-2). Testing of 187 samples showed high fecal indicator bacteria concentrations at both up- and downstream sites. Overall, 40%, 23%, and 61% of samples exceeded state and federal recreational water quality guidelines for fecal coliforms, E. coli, and Enterococcus, respectively. Pig-1-Bac and Pig-2-Bac showed the highest specificity to swine fecal wastes and were 2.47 (95% confidence interval [CI]=1.03, 5.94) and 2.30 times (95% CI=0.90, 5.88) as prevalent proximal down- than proximal upstream of swine CAFOs, respectively. Pig-1-Bac and Pig-2-Bac were also 2.87 (95% CI=1.21, 6.80) and 3.36 (95% CI=1.34, 8.41) times as prevalent when 48 hour antecedent rainfall was greater than versus less than the mean, respectively. Results suggest diffuse and overall poor sanitary quality of surface waters where swine CAFO density is high. Pig-1-Bac and Pig-2-Bac are useful for tracking off-site conveyance of swine fecal wastes into surface waters proximal to and downstream of swine CAFOs and during rain events. Copyright © 2014 Elsevier B.V. All rights reserved.

  8. Fecal indicator bacteria contamination of fomites and household demand for surface disinfection products: a case study from Peru.

    PubMed

    Julian, Timothy R; MacDonald, Luke H; Guo, Yayi; Marks, Sara J; Kosek, Margaret; Yori, Pablo P; Pinedo, Silvia Rengifo; Schwab, Kellogg J

    2013-11-01

    Surface-mediated disease transmission is understudied in developing countries, particularly in light of the evidence that surface concentrations of fecal bacteria typically exceed concentrations in developed countries by 10- to 100-fold. In this study, we examined fecal indicator bacterial contamination of dinner plates at 21 households in four peri-urban communities in the Peruvian Amazon. We also used surveys to estimate household use of and demand for surface disinfectants at 280 households. Despite detecting total coliform, enterococci, and Escherichia coli on 86%, 43%, and 24% of plates sampled, respectively, less than one-third of households were regularly using bleach to disinfect surfaces. Among non-users of bleach, only 3.2% of respondents reported a new demand for bleach, defined as a high likelihood of using bleach within the next year. This study highlights the potential for marketing approaches to increase use of and demand for surface disinfectants to improve domestic hygiene.

  9. Fecal Indicator Bacteria Contamination of Fomites and Household Demand for Surface Disinfection Products: A Case Study from Peru

    PubMed Central

    Julian, Timothy R.; MacDonald, Luke H.; Guo, Yayi; Marks, Sara J.; Kosek, Margaret; Yori, Pablo P.; Pinedo, Silvia Rengifo; Schwab, Kellogg J.

    2013-01-01

    Surface-mediated disease transmission is understudied in developing countries, particularly in light of the evidence that surface concentrations of fecal bacteria typically exceed concentrations in developed countries by 10- to 100-fold. In this study, we examined fecal indicator bacterial contamination of dinner plates at 21 households in four peri-urban communities in the Peruvian Amazon. We also used surveys to estimate household use of and demand for surface disinfectants at 280 households. Despite detecting total coliform, enterococci, and Escherichia coli on 86%, 43%, and 24% of plates sampled, respectively, less than one-third of households were regularly using bleach to disinfect surfaces. Among non-users of bleach, only 3.2% of respondents reported a new demand for bleach, defined as a high likelihood of using bleach within the next year. This study highlights the potential for marketing approaches to increase use of and demand for surface disinfectants to improve domestic hygiene. PMID:24019431

  10. High-throughput DNA sequence analysis reveals stable engraftment of gut microbiota following transplantation of previously frozen fecal bacteria.

    PubMed

    Hamilton, Matthew J; Weingarden, Alexa R; Unno, Tatsuya; Khoruts, Alexander; Sadowsky, Michael J

    2013-01-01

    Fecal microbiota transplantation (FMT) is becoming a more widely used technology for treatment of recurrent Clostridum difficile infection (CDI). While previous treatments used fresh fecal slurries as a source of microbiota for FMT, we recently reported the successful use of standardized, partially purified and frozen fecal microbiota to treat CDI. Here we report that high-throughput 16S rRNA gene sequencing showed stable engraftment of gut microbiota following FMT using frozen fecal bacteria from a healthy donor. Similar bacterial taxa were found in post-transplantation samples obtained from the recipients and donor samples, but the relative abundance varied considerably between patients and time points. Post FMT samples from patients showed an increase in the abundance of Firmicutes and Bacteroidetes, representing 75-80% of the total sequence reads. Proteobacteria and Actinobacteria were less abundant (< 5%) than that found in patients prior to FMT. Post FMT samples from two patients were very similar to donor samples, with the Bacteroidetes phylum represented by a great abundance of members of the families Bacteroidaceae, Rikenellaceae and Porphyromonadaceae, and were largely comprised of Bacteroides, Alistipes and Parabacteroides genera. Members of the phylum Firmicutes were represented by Ruminococcaceae, Lachnospiraceae, Verrucomicrobiaceae and unclassified Clostridiales and members of the Firmicutes. One patient subsequently received antibiotics for an unrelated infection, resulting in an increase in the number of intestinal Proteobacteria, primarily Enterobacteriaceae. Our results demonstrate that frozen fecal microbiota from a healthy donor can be used to effectively treat recurrent CDI resulting in restoration of the structure of gut microbiota and clearing of Clostridum difficile.

  11. Comparison of Sewage and Animal Fecal Microbiomes by Using Oligotyping Reveals Potential Human Fecal Indicators in Multiple Taxonomic Groups

    PubMed Central

    Fisher, Jenny C.; Eren, A. Murat; Green, Hyatt C.; Shanks, Orin C.; Morrison, Hilary G.; Vineis, Joseph H.; Sogin, Mitchell L.

    2015-01-01

    Most DNA-based microbial source tracking (MST) approaches target host-associated organisms within the order Bacteroidales, but the gut microbiota of humans and other animals contain organisms from an array of other taxonomic groups that might provide indicators of fecal pollution sources. To discern between human and nonhuman fecal sources, we compared the V6 regions of the 16S rRNA genes detected in fecal samples from six animal hosts to those found in sewage (as a proxy for humans). We focused on 10 abundant genera and used oligotyping, which can detect subtle differences between rRNA gene sequences from ecologically distinct organisms. Our analysis showed clear patterns of differential oligotype distributions between sewage and animal samples. Over 100 oligotypes of human origin occurred preferentially in sewage samples, and 99 human oligotypes were sewage specific. Sequences represented by the sewage-specific oligotypes can be used individually for development of PCR-based assays or together with the oligotypes preferentially associated with sewage to implement a signature-based approach. Analysis of sewage from Spain and Brazil showed that the sewage-specific oligotypes identified in U.S. sewage have the potential to be used as global alternative indicators of human fecal pollution. Environmental samples with evidence of prior human fecal contamination had consistent ratios of sewage signature oligotypes that corresponded to the trends observed for sewage. Our methodology represents a promising approach to identifying new bacterial taxa for MST applications and further highlights the potential of the family Lachnospiraceae to provide human-specific markers. In addition to source tracking applications, the patterns of the fine-scale population structure within fecal taxa suggest a fundamental relationship between bacteria and their hosts. PMID:26231648

  12. Effects of feeding elevated concentrations of copper and zinc on the antimicrobial susceptibilities of fecal bacteria in feedlot cattle.

    PubMed

    Jacob, Megan E; Fox, J Trent; Nagaraja, T G; Drouillard, James S; Amachawadi, Raghavendra G; Narayanan, Sanjeev K

    2010-06-01

    Cattle are fed elevated concentrations of copper and zinc for growth promotion. The potential mechanisms of growth promotional effects of these elements are attributed to their antimicrobial activities, similar to that of antibiotics, in that gut microbial flora are altered to reduce fermentation loss of nutrients and to suppress gut pathogens. Copper and zinc fed at elevated concentrations may select for bacteria that are resistant not only to heavy metals but also to antibiotics. Our objectives were to determine the effects of feeding elevated copper and zinc on the antimicrobial susceptibilities of fecal bacteria in feedlot cattle. Twenty heifers, fed corn-based high-grain diets, were randomly assigned to treatments in a 2 x 2 factorial arrangement with 1X or 10X National Research Council recommended copper and/or zinc. Feces, collected on days 0, 14, and 32, were cultured for commensal bacteria (Escherichia coli and Enterococcus) to determine their susceptibilities to copper, zinc, and antibiotics. Fecal DNA was extracted to detect tcrB gene and quantify erm(B) and tet(M) genes. In E. coli and Enterococcus sp., minimal differences in minimum inhibitory concentrations (MICs) of copper, zinc, and antibiotics were noticed. The mean copper MIC for E. coli increased (p < 0.05) between days 0 and 32 and days 14 and 32. The only treatment effect detected was increased zinc MIC of E. coli isolates (p < 0.01). The tcrB gene was not detected in feces or in enterococcal isolates. Proportions of erm(B) and tet(M) were unaffected by copper or zinc supplementation. However, the proportion of tet(M) increased (p < 0.05) between days 0 and 14. Feeding elevated copper and/or zinc to feedlot cattle had marginal effects on antimicrobial susceptibilities of fecal E. coli and enterococci.

  13. Sunlight Inactivation of Fecal Bacteriophages and Bacteria in Sewage-Polluted Seawater

    PubMed Central

    Sinton, Lester W.; Finlay, Rochelle K.; Lynch, Philippa A.

    1999-01-01

    Sunlight inactivation rates of somatic coliphages, F-specific RNA bacteriophages (F-RNA phages), and fecal coliforms were compared in seven summer and three winter survival experiments. Experiments were conducted outdoors, using 300-liter 2% (vol/vol) sewage-seawater mixtures held in open-top chambers. Dark inactivation rates (kDs), measured from exponential survival curves in enclosed (control) chambers, were higher in summer (temperature range: 14 to 20°C) than in winter (temperature range: 8 to 10°C). Winter kDs were highest for fecal coliforms and lowest for F-RNA phages but were the same or similar for all three indicators in summer. Sunlight inactivation rates (kS), as a function of cumulative global solar radiation (insolation), were all higher than the kDs with a consistent kS ranking (from greatest to least) as follows: fecal coliforms, F-RNA phages, and somatic coliphages. Phage inactivation was exponential, but bacterial curves typically exhibited a shoulder. Phages from raw sewage exhibited kSs similar to those from waste stabilization pond effluent, but raw sewage fecal coliforms were inactivated faster than pond effluent fecal coliforms. In an experiment which included F-DNA phages and Bacteroides fragilis phages, the kS ranking (from greatest to least) was as follows: fecal coliforms, F-RNA phages, B. fragilis phages, F-DNA phages, and somatic coliphages. In a 2-day experiment which included enterococci, the initial concentration ranking (from greatest to least: fecal coliforms, enterococci, F-RNA phages, and somatic coliphages) was reversed during sunlight exposure, with only the phages remaining detectable by the end of day 2. Inactivation rates under different optical filters decreased with the increase in spectral cutoff wavelength (50% light transmission) and indicated that F-RNA phages and fecal coliforms are more susceptible than somatic coliphages to longer solar wavelengths, which predominate in seawater. The consistently superior survival

  14. Spatial and hydrologic variation of Bacteroidales, adenovirus and enterovirus in a semi-arid, wastewater effluent-impacted watershed.

    PubMed

    Bambic, Dustin G; Kildare-Hann, Beverly J; Rajal, Veronica B; Sturm, Belinda S M; Minton, Chris B; Schriewer, Alexander; Wuertz, Stefan

    2015-05-15

    Bacteroidales and viruses were contemporaneously measured during dry and wet weather conditions at a watershed-scale in a semi-arid watershed impacted by a mixture of agricultural runoff, municipal wastewater effluent and municipal runoff. The results highlight the presence of municipal wastewater effluent as a confounding factor for microbial source tracking (MST) studies, and thus data were segregated into groups based on whether they were impacted by wastewater effluent. In semi-arid environments such as the Calleguas Creek watershed, located in southern California, the relative contribution of municipal wastewater effluent is dependent on hydrology as storm events lead to conditions where agricultural and municipal stormwater dominate receiving waters (rather than municipal wastewater, which is the case during dry weather). As such, the approach to data segregation was dependent on hydrology/storm conditions. Storm events led to significant increases in ruminant- and dog-associated Bacteroidales concentrations, indicating that overland transport connects strong non-human fecal sources with surface waters. Because the dataset had a large number of non-detect samples, data handling included the Kaplan-Meir estimator and data were presented graphically in a manner that reflects the potential effect of detection limits. In surface water samples with virus detections, Escherichia coli concentrations were often below (in compliance with) the recreational water quality criteria. In fact, sites downstream of direct inputs of municipal wastewater effluent exhibited the lowest concentrations of E. coli, but the highest concentrations of human-associated Bacteroidales and highest detection rates of human viruses. The toolkit, comprised of the four Bacteroidales assays and human virus assays used, can be successfully applied to inform watershed managers seeking to comply with recreational water quality criteria. However, care should be taken when analyzing data to account

  15. Longitudinal Poisson regression to evaluate the epidemiology of Cryptosporidium, Giardia, and fecal indicator bacteria in coastal California wetlands.

    PubMed

    Hogan, Jennifer N; Daniels, Miles E; Watson, Fred G; Conrad, Patricia A; Oates, Stori C; Miller, Melissa A; Hardin, Dane; Byrne, Barbara A; Dominik, Clare; Melli, Ann; Jessup, David A; Miller, Woutrina A

    2012-05-01

    Fecal pathogen contamination of watersheds worldwide is increasingly recognized, and natural wetlands may have an important role in mitigating fecal pathogen pollution flowing downstream. Given that waterborne protozoa, such as Cryptosporidium and Giardia, are transported within surface waters, this study evaluated associations between fecal protozoa and various wetland-specific and environmental risk factors. This study focused on three distinct coastal California wetlands: (i) a tidally influenced slough bordered by urban and agricultural areas, (ii) a seasonal wetland adjacent to a dairy, and (iii) a constructed wetland that receives agricultural runoff. Wetland type, seasonality, rainfall, and various water quality parameters were evaluated using longitudinal Poisson regression to model effects on concentrations of protozoa and indicator bacteria (Escherichia coli and total coliform). Among wetland types, the dairy wetland exhibited the highest protozoal and bacterial concentrations, and despite significant reductions in microbe concentrations, the wetland could still be seen to influence water quality in the downstream tidal wetland. Additionally, recent rainfall events were associated with higher protozoal and bacterial counts in wetland water samples across all wetland types. Notably, detection of E. coli concentrations greater than a 400 most probable number (MPN) per 100 ml was associated with higher Cryptosporidium oocyst and Giardia cyst concentrations. These findings show that natural wetlands draining agricultural and livestock operation runoff into human-utilized waterways should be considered potential sources of pathogens and that wetlands can be instrumental in reducing pathogen loads to downstream waters.

  16. Longitudinal Poisson Regression To Evaluate the Epidemiology of Cryptosporidium, Giardia, and Fecal Indicator Bacteria in Coastal California Wetlands

    PubMed Central

    Hogan, Jennifer N.; Daniels, Miles E.; Watson, Fred G.; Conrad, Patricia A.; Oates, Stori C.; Miller, Melissa A.; Hardin, Dane; Byrne, Barbara A.; Dominik, Clare; Melli, Ann; Jessup, David A.

    2012-01-01

    Fecal pathogen contamination of watersheds worldwide is increasingly recognized, and natural wetlands may have an important role in mitigating fecal pathogen pollution flowing downstream. Given that waterborne protozoa, such as Cryptosporidium and Giardia, are transported within surface waters, this study evaluated associations between fecal protozoa and various wetland-specific and environmental risk factors. This study focused on three distinct coastal California wetlands: (i) a tidally influenced slough bordered by urban and agricultural areas, (ii) a seasonal wetland adjacent to a dairy, and (iii) a constructed wetland that receives agricultural runoff. Wetland type, seasonality, rainfall, and various water quality parameters were evaluated using longitudinal Poisson regression to model effects on concentrations of protozoa and indicator bacteria (Escherichia coli and total coliform). Among wetland types, the dairy wetland exhibited the highest protozoal and bacterial concentrations, and despite significant reductions in microbe concentrations, the wetland could still be seen to influence water quality in the downstream tidal wetland. Additionally, recent rainfall events were associated with higher protozoal and bacterial counts in wetland water samples across all wetland types. Notably, detection of E. coli concentrations greater than a 400 most probable number (MPN) per 100 ml was associated with higher Cryptosporidium oocyst and Giardia cyst concentrations. These findings show that natural wetlands draining agricultural and livestock operation runoff into human-utilized waterways should be considered potential sources of pathogens and that wetlands can be instrumental in reducing pathogen loads to downstream waters. PMID:22427504

  17. Occurrence and distribution of fecal indicator bacteria and gene markers of pathogenic bacteria in Great Lakes tributaries, March-October 2011

    USGS Publications Warehouse

    Brennan, Angela K.; Johnson, Heather E.; Totten, Alexander R.; Duris, Joseph W.

    2015-01-01

    Water samples were analyzed for the presence of FIB concentrations (FIB; fecal coliform bacteria, Escherichia coli [E. coli], and enterococci) by using membrane filtration and serial dilution methods. The resulting enrichments from standard culturing of the samples were then analyzed by using polymerase chain reaction (PCR) to determine the occurrence of pathogen gene markers for Shigella species, Campylobacter jejuni and coli, Salmonellaspecies, and pathogenic E. coli, including Shiga toxin-producing E. coli (STEC).

  18. Development and Testing of Novel Canine Fecal Source-Identification Assays

    EPA Science Inventory

    The extent to which dogs contribute to aquatic fecal contamination is unknown despite the potential for zoonotic transfer of harmful human pathogens. Recent method comparison studies have shown that available Bacteroidales 16S rRNA-based methods for the detection of canine fecal ...

  19. Development and Testing of Novel Canine Fecal Source-Identification Assays

    EPA Science Inventory

    The extent to which dogs contribute to aquatic fecal contamination is unknown despite the potential for zoonotic transfer of harmful human pathogens. Recent method comparison studies have shown that available Bacteroidales 16S rRNA-based methods for the detection of canine fecal ...

  20. Characterization of fecal indicator bacteria in sediments cores from the largest freshwater lake of Western Europe (Lake Geneva, Switzerland).

    PubMed

    Thevenon, Florian; Regier, Nicole; Benagli, Cinzia; Tonolla, Mauro; Adatte, Thierry; Wildi, Walter; Poté, John

    2012-04-01

    This study characterized the fecal indicator bacteria (FIB), including Escherichia coli (E. coli) and Enteroccocus (ENT), disseminated over time in the Bay of Vidy, which is the most contaminated area of Lake Geneva. Sediments were collected from a site located at ∼500 m from the present waste water treatment plant (WWTP) outlet pipe, in front of the former WWTP outlet pipe, which was located at only 300 m from the coastal recreational area (before 2001). E. coli and ENT were enumerated in sediment suspension using the membrane filter method. The FIB characterization was performed for human Enterococcus faecalis (E. faecalis) and Enterococcus faecium (E. faecium) and human specific bacteroides by PCR using specific primers and a matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Bacterial cultures revealed that maximum values of 35.2 × 10(8) and 6.6 × 10(6)CFU g(-1) dry sediment for E. coli and ENT, respectively, were found in the sediments deposited following eutrophication of Lake Geneva in the 1970s, whereas the WWTP started operating in 1964. The same tendency was observed for the presence of human fecal pollution: the percentage of PCR amplification with primers ESP-1/ESP-2 for E. faecalis and E. faecium indicated that more than 90% of these bacteria were from human origin. Interestingly, the PCR assays for specific-human bacteroides HF183/HF134 were positive for DNA extracted from all isolated strains of sediment surrounding WWPT outlet pipe discharge. The MALDI-TOF MS confirmed the presence of general E. coli and predominance E. faecium in isolated strains. Our results demonstrated that human fecal bacteria highly increased in the sediments contaminated with WWTP effluent following the eutrophication of Lake Geneva. Additionally, other FIB cultivable strains from animals or adapted environmental strains were detected in the sediment of the bay. The approaches used in this research are valuable to assess the

  1. FECAL BACTERIA SOURCE TRACKING AND BACTEROIDES SPP. HOST SPECIES SPECIFICITY ANALYSIS

    EPA Science Inventory

    Point and non-point pollution sources of fecal pollution on a watershed adversely impact the quality of drinking source waters and recreational waters. States are required to develop total maximum daily loads (TMDLs) and devise best management practices (BMPs) to reduce the po...

  2. Influence of weaning on fecal shedding of pathogenic bacteria in dairy calves

    USDA-ARS?s Scientific Manuscript database

    The objectives of the current research were to determine the effect of weaning on fecal shedding of Salmonella and E. coli O157:H7 in dairy calves and to examine cultured isolates (to include Enterococcus) for antimicrobial susceptibility. This research was conducted on one large commercial dairy (...

  3. Distribution of Genetic Marker Concentrations for Fecal Indicator Bacteria in Sewage and Animal Feces

    EPA Science Inventory

    The application of quantitative real-time PCR (qPCR) methods for the identification of fecal microorganisms in surface waters has the potential to revolutionize water quality monitoring worldwide. Unlike traditional cultivation methods, qPCR estimates the concentration of gen...

  4. FECAL BACTERIA SOURCE TRACKING AND BACTEROIDES SPP. HOST SPECIES SPECIFICITY ANALYSIS

    EPA Science Inventory

    Point and non-point pollution sources of fecal pollution on a watershed adversely impact the quality of drinking source waters and recreational waters. States are required to develop total maximum daily loads (TMDLs) and devise best management practices (BMPs) to reduce the po...

  5. Distribution of Genetic Marker Concentrations for Fecal Indicator Bacteria in Sewage and Animal Feces

    EPA Science Inventory

    The application of quantitative real-time PCR (qPCR) methods for the identification of fecal microorganisms in surface waters has the potential to revolutionize water quality monitoring worldwide. Unlike traditional cultivation methods, qPCR estimates the concentration of gen...

  6. Within-House Spatial Distribution of Fecal Indicator Bacteria in Poultry Litter

    USDA-ARS?s Scientific Manuscript database

    Land application of poultry litter is often considered to be a major source of water pollutants in poultry-producing regions. However, reported levels of fecal indicator microorganisms in litter vary widely with considerable variation possible within houses and across farms depending upon management...

  7. Engineering solutions to improve the removal of fecal indicator bacteria by bioinfiltration systems during intermittent flow of stormwater.

    PubMed

    Mohanty, Sanjay K; Torkelson, Andrew A; Dodd, Hanna; Nelson, Kara L; Boehm, Alexandria B

    2013-10-01

    Bioinfiltration systems facilitate the infiltration of urban stormwater into soil and reduce high flow events and flooding. Stormwater carries a myriad of pollutants including fecal indicator bacteria (FIB). Significant knowledge gaps exist about the ability of bioinfiltration systems to remove and retain FIB. The present study investigates the ability of model, simplified bioinfiltration systems containing quartz sand and iron oxide-coated quartz sand (IOCS) to remove two FIB (Enterococcus faecalis and Escherichia coli) suspended in synthetic stormwater with and without natural organic matter (NOM) as well as the potential for accumulated FIB to be remobilized during intermittent flow. The experiments were conducted in two phases: (1) the saturated columns packed with either sand or IOCS were contaminated by injecting stormwater with bacteria followed by injection of sterile stormwater and (2) the contaminated columns were subjected to intermittent infiltration of sterile stormwater preceded by a pause during which columns were either kept saturated or drained by gravity. During intermittent flow, fewer bacteria were released from the saturated column compared to the column drained by gravity: 12% of attached E. coli and 3% of attached Ent. faecalis were mobilized from the drained sand column compared to 3% of attached E. coli and 2% attached Ent. faecalis mobilized from the saturated sand column. Dry and wet cycles introduce moving air-water interfaces that can scour bacteria from grain surfaces. During intermittent flows, less than 0.2% of attached bacteria were mobilized from IOCS, which bound both bacteria irreversibly in the absence of NOM. Addition of NOM, however, increased bacterial mobilization from IOCS: 50% of attached E. coli and 8% of attached Ent. faecalis were released from IOCS columns during draining and rewetting. Results indicate that using geomedia such as IOCS that promote irreversible attachment of bacteria, and maintaining saturated

  8. Measuring and modeling the flux of fecal bacteria across the sediment-water interface in a turbulent stream

    NASA Astrophysics Data System (ADS)

    Grant, Stanley B.; Litton-Mueller, Rachel M.; Ahn, Jong H.

    2011-05-01

    Sediments are a pervasive source of fecal indicator bacteria (FIB) in rivers, lakes, estuaries, and oceans and may constitute a long-term reservoir of human disease. Previous attempts to quantify the flux of FIB across the sediment-water interface (SWI) are limited to extreme flow events, for which the primary mechanism of bacterial release is disruption and/or erosion of the sediment substrate. Here we report measurements of FIB flux across the SWI in a turbulent stream that is not undergoing significant erosion. The stream is formed by the steady discharge of bacteria-free disinfected and highly treated wastewater effluent to an earthen channel harboring high concentrations of FIB in the sediment from in situ growth. The flux j″ of FIB across the SWI, estimated from mass balance on FIB measurements in the water column, scales linearly with the concentration of bacteria in sediment pore fluids Cpore over a 3 decade change in both variables: ? The magnitude of the observed mass transfer velocity (? m s-1) is significantly larger than values predicted for either the diffusion of bacteria across a concentration boundary layer (? m s-1) or sweep and eject fluid motions at the SWI (? m s-1) but is similar to the flux of water between the stream and its hyporheic zone estimated from dye injection experiments. These results support the hypothesis that hyporheic exchange controls the trafficking of bacteria, and perhaps other types of particulate organic matter, across the SWI in turbulent streams.

  9. Characterization of the fecal bacteria communities of forage-fed horses by pyrosequencing of 16S rRNA V4 gene amplicons.

    PubMed

    Shepherd, Megan L; Swecker, William S; Jensen, Roderick V; Ponder, Monica A

    2012-01-01

    The diversity of the equine fecal bacterial community was evaluated using pyrosequencing of 16S rRNA gene amplicons. Fecal samples were obtained from horses fed cool-season grass hay. Fecal bacteria were characterized by amplifying the V4 region of bacterial 16S rRNA gene. Of 5898 mean unique sequences, a mean of 1510 operational taxonomic units were identified in the four fecal samples. Equine fecal bacterial richness was higher than that reported in humans, but lower than that reported in either cattle feces or soil. Bacterial classified sequences were assigned to 16 phyla, of which 10 were present in all samples. The largest number of reads belonged to Firmicutes (43.7% of total bacterial sequences), Verrucomicrobia (4.1%), Proteobacteria (3.8%), and Bacteroidetes (3.7%). The less abundant Actinobacteria, Cyanobacteria, and TM7 phyla presented here have not been previously described in the gut contents or feces of horses. Unclassified sequences represented 38.1% of total bacterial sequences; therefore, the equine fecal microbiome diversity is likely greater than that described. This is the first study to characterize the fecal bacterial community in horses by the use of 16S rRNA gene amplicon pyrosequencing, expanding our knowledge of the fecal microbiota of forage-fed horses.

  10. Comparison of two methods for detection of fecal indicator bacteria used in water quality monitoring of the Three Gorges Reservoir.

    PubMed

    Wang, Zhaodan; Xiao, Guosheng; Zhou, Nong; Qi, Wenhua; Han, Lin; Ruan, Yu; Guo, Dongqin; Zhou, Hong

    2015-12-01

    Scientifically sound methods to rapidly measure fecal indicator bacteria are important to ensure safe water for drinking and recreational purposes. A total of 200 water samples obtained from the Three Gorges Reservoir during three successive one-year study periods (October 2009 to September 2012) were analyzed using multiple-tube fermentation (MTF) and most probable numbers combined with polymerase chain reaction (MPN-PCR). The MPN-PCR method was found to be significantly more sensitive than the MTF method for detecting Escherichia coli and Enterococcus spp., and of equal sensitivity for detecting total coliforms when all surface water samples were grouped together. The two analytical methods had a strong, significant relationship, but MPN-PCR took only 12-18hr, compared with the 3-8days needed using the MTF method. Bacterial concentrations varied per sampling site but were significantly lower in the mainstream of the Yangtze River than those in the backwater areas of tributaries. The water quality of 85.8% of water samples from the mainstream was suitable for use as a centralized potable water source, while the water quality of 52.5% of water samples from the backwater areas was unsuitable for recreational activities. Relationships between fecal indicator bacteria showed significant correlation (r=0.636-0.909, p<0.01, n=200), while a weak but significant correlation was found between fecal indicators and water turbidity, water temperature, daily inflow, and total dissolved solids (r=0.237-0.532, p<0.05, n=200). The study indicated that MPN-PCR is a rapid and easily performed deoxyribonucleic acid (DNA)-based method for quantitative detection of viable total coliforms, E. coli, and Enterococcus spp. in surface water.

  11. Linking fecal bacteria in rivers to landscape, geochemical, and hydrologic factors and sources at the basin scale

    PubMed Central

    Verhougstraete, Marc P.; Martin, Sherry L.; Kendall, Anthony D.; Hyndman, David W.; Rose, Joan B.

    2015-01-01

    Linking fecal indicator bacteria concentrations in large mixed-use watersheds back to diffuse human sources, such as septic systems, has met limited success. In this study, 64 rivers that drain 84% of Michigan’s Lower Peninsula were sampled under baseflow conditions for Escherichia coli, Bacteroides thetaiotaomicron (a human source-tracking marker), landscape characteristics, and geochemical and hydrologic variables. E. coli and B. thetaiotaomicron were routinely detected in sampled rivers and an E. coli reference level was defined (1.4 log10 most probable number⋅100 mL−1). Using classification and regression tree analysis and demographic estimates of wastewater treatments per watershed, septic systems seem to be the primary driver of fecal bacteria levels. In particular, watersheds with more than 1,621 septic systems exhibited significantly higher concentrations of B. thetaiotaomicron. This information is vital for evaluating water quality and health implications, determining the impacts of septic systems on watersheds, and improving management decisions for locating, constructing, and maintaining on-site wastewater treatment systems. PMID:26240328

  12. Linking fecal bacteria in rivers to landscape, geochemical, and hydrologic factors and sources at the basin scale.

    PubMed

    Verhougstraete, Marc P; Martin, Sherry L; Kendall, Anthony D; Hyndman, David W; Rose, Joan B

    2015-08-18

    Linking fecal indicator bacteria concentrations in large mixed-use watersheds back to diffuse human sources, such as septic systems, has met limited success. In this study, 64 rivers that drain 84% of Michigan's Lower Peninsula were sampled under baseflow conditions for Escherichia coli, Bacteroides thetaiotaomicron (a human source-tracking marker), landscape characteristics, and geochemical and hydrologic variables. E. coli and B. thetaiotaomicron were routinely detected in sampled rivers and an E. coli reference level was defined (1.4 log10 most probable number⋅100 mL(-1)). Using classification and regression tree analysis and demographic estimates of wastewater treatments per watershed, septic systems seem to be the primary driver of fecal bacteria levels. In particular, watersheds with more than 1,621 septic systems exhibited significantly higher concentrations of B. thetaiotaomicron. This information is vital for evaluating water quality and health implications, determining the impacts of septic systems on watersheds, and improving management decisions for locating, constructing, and maintaining on-site wastewater treatment systems.

  13. Conclusions and future use of fecal indicator bacteria for monitoring water quality and protecting human health

    USGS Publications Warehouse

    Sadowsky, Michael J.; Whitman, Richard L.

    2011-01-01

    A summary of the focus and the recurring theme of the book is presented in this chapter. It includes the use of faecal bacteria as an indicator of faecal pollution and water quality, ubiquity of faecal bacteria, and sources and movement of faecal bacteria in the environment.

  14. Survival of Host-Associated Bacteroidales Cells and Their Relationship with Enterococcus spp., Campylobacter jejuni, Salmonella enterica Serovar Typhimurium, and Adenovirus in Freshwater Microcosms as Measured by Propidium Monoazide-Quantitative PCR

    PubMed Central

    Bae, Sungwoo

    2012-01-01

    The ideal host-associated genetic fecal marker would be capable of predicting the presence of specific pathogens of concern. Flowthrough freshwater microcosms containing mixed feces and inocula of the pathogens Campylobacter jejuni, Salmonella enterica serovar Typhimurium, and adenovirus were placed at ambient temperature in the presence and absence of diurnal sunlight. The total Enterococcus DNA increased during the early periods (23 h) under sunlight exposure, even though cultivable Enterococcus and DNA in intact cells, as measured by propidium monoazide (PMA), decreased with first-order kinetics during the entire period. We found a significant difference in the decay of host-associated Bacteroidales cells between sunlight exposure and dark conditions (P value < 0.05), whereas the persistence of host-associated Bacteroidales DNA was comparable. The 2-log reduction times of adenovirus were 72 h for sunlight exposure and 99 h for dark conditions with similar decay rate constants (P value = 0.13). The persistences of fecal Bacteroidales cells and Campylobacter cells exposed to sunlight were similar, and host-associated Bacteroidales DNA and waterborne pathogen DNA were degraded at comparable rates (P values > 0.05). Overall, the ratio of quantitative PCR (qPCR) cycle threshold (CT) values with and without PMA treatment was indicative of the time elapsed since inoculation of the microcosm with (i) fecal material from different animal sources based on host-associated Bacteroidales and (ii) pure cultures of bacterial pathogens. The use of both PMA-qPCR and qPCR may yield more realistic information about recent sources of fecal contamination and result in improved prediction of waterborne pathogens and assessment of health risk. PMID:22139002

  15. A BASIN-WIDE ANALYSIS OF THE DYNAMICS OF FECAL CONTAMINATION AND FECAL SOURCE IDENTIFICATION IN TILLAMOOK BAY, OREGON

    EPA Science Inventory

    The objectives of this study were to determine if spatial and temporal dynamics exist in source-specific Bacteroidales 16S rRNA genetic marker data across a watershed, to study these in relation to fecal indicator counts, general measurements of water quality, and climat...

  16. A BASIN-WIDE ANALYSIS OF THE DYNAMICS OF FECAL CONTAMINATION AND FECAL SOURCE IDENTIFICATION IN TILLAMOOK BAY, OREGON

    EPA Science Inventory

    The objectives of this study were to determine if spatial and temporal dynamics exist in source-specific Bacteroidales 16S rRNA genetic marker data across a watershed, to study these in relation to fecal indicator counts, general measurements of water quality, and climat...

  17. Comparison of Sewage and Animal Fecal Microbiomes by using Oligotyping Reveals Potential Human Fecal Indicators in Multiple Taxonomic Groups

    EPA Science Inventory

    Most DNA-based microbial source tracking (MST) approaches target host-associated organisms within the order Bacteroidales, but human and other animal gut microbiota contain an array of other taxonomic groups that might serve as indicators for sources of fecal pollution. High thr...

  18. Comparison of Sewage and Animal Fecal Microbiomes by using Oligotyping Reveals Potential Human Fecal Indicators in Multiple Taxonomic Groups

    EPA Science Inventory

    Most DNA-based microbial source tracking (MST) approaches target host-associated organisms within the order Bacteroidales, but human and other animal gut microbiota contain an array of other taxonomic groups that might serve as indicators for sources of fecal pollution. High thr...

  19. Occurrence of fecal indicator bacteria in surface waters and the subsurface aquifer in Key Largo, Florida.

    PubMed Central

    Paul, J H; Rose, J B; Jiang, S; Kellogg, C; Shinn, E A

    1995-01-01

    Sewage waste disposal facilities in the Florida Keys include septic tanks and individual package plants in place of municipal collection facilities in most locations. In Key Largo, both facilities discharge into the extremely porous Key Largo limestone. To determine whether there was potential contamination of the subsurface aquifer and nearby coastal surface waters by such waste disposal practices, we examined the presence of microbial indicators commonly found in sewage (fecal coliforms, Clostridium perfringens, and enterococci) and aquatic microbial parameters (viral direct counts, bacterial direct counts, chlorophyll a, and marine vibriophage) in injection well effluent, monitoring wells that followed a transect from onshore to offshore, and surface waters above these wells in two separate locations in Key Largo in August 1993 and March 1994. Effluent and waters from onshore shallow monitoring wells (1.8- to 3.7-m depth) contained two or all three of the fecal indicators in all three samples taken, whereas deeper wells (10.7- to 12.2-m depth) at these same sites contained few or none. The presence of fecal indicators was found in two of five nearshore wells (i.e., those that were < or = 1.8 miles [< or = 2.9 km] from shore), whereas offshore wells (> or = 2.1 to 5.7 miles [< or = 3.4 to 9.2 km] from shore) showed little sign of contamination. Indicators were also found in surface waters in a canal in Key Largo and in offshore surface waters in March but not in August. Collectively, these results suggest that fecal contamination of the shallow onshore aquifer, parts of the nearshore aquifer, and certain surface waters has occurred.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7793943

  20. A Synoptic Study of Fecal-Indicator Bacteria in the Wind River, Bighorn River, and Goose Creek Basins, Wyoming, June-July 2000

    USGS Publications Warehouse

    Clark, Melanie L.; Gamper, Merry E.

    2003-01-01

    A synoptic study of fecal-indicator bacteria was conducted during June and July 2000 in the Wind River, Bighorn River, and Goose Creek Basins in Wyoming as part of the U.S. Geological Survey's National Water-Quality Assessment Program for the Yellowstone River Basin. Fecal-coliform concentrations ranged from 2 to 3,000 col/100 mL (colonies per 100 milliliters) for 100 samples, and Escherichia coli concentrations ranged from 1 to 2,800 col/100 mL for 97 samples. Fecal-coliform concentrations exceeded the U.S. Environmental Protection Agency's recommended limit for a single sample for recreational contact with water in 37.0 percent of the samples. Escherichia coli concentrations exceeded the U.S. Environmental Protection Agency's recommended limit for a single sample for moderate use, full-body recreational contact with water in 38.1 percent of the samples and the recommended limit for infrequent use, full-body recreational contact with water in 24.7 percent of the samples. Fecal-indicator-bacteria concentrations varied by basin. Samples from the Bighorn River Basin had the highest median concentrations for fecal coliform of 340 col/100 mL and for Escherichia coli of 300 col/100 mL. Samples from the Wind River Basin had the lowest median concentrations for fecal coliform of 50 col/100 mL and for Escherichia coli of 62 col/100 mL. Fecal-indicator-bacteria concentrations varied by land cover. Samples from sites with an urban land cover had the highest median concentrations for fecal coliform of 540 col/100 mL and for Escherichia coli of 420 col/100 mL. Maximum concentrations for fecal coliform of 3,000 col/100 mL and for Escherichia coli of 2,800 col/100 mL were in samples from sites with an agricultural land cover. The lowest median concentrations for fecal coliform of 130 col/100 mL and for Escherichia coli of 67 col/100 mL were for samples from sites with a forested land cover. A strong and positive relation existed between fecal coliform and Escherichia coli

  1. Modeling the dry-weather tidal cycling of fecal indicator bacteria in surface waters of an intertidal wetland.

    PubMed

    Sanders, Brett F; Arega, Feleke; Sutula, Martha

    2005-09-01

    Recreational water quality at beaches in California and elsewhere is often poor near the outlets of rivers, estuaries, and lagoons. This condition has prompted interest in the role of wetlands in modulating surface water concentrations of fecal indicator bacteria (FIB), the basis of water quality standards internationally. A model was developed and applied to predict the dry-weather tidal cycling of FIB in Talbert Marsh, an estuarine, intertidal wetland in Huntington Beach, California, in response to loads from urban runoff, bird feces, and resuspended sediments. The model predicts the advection, dispersion and die-off of total coliform, Escherichia coli, and enterococci using a depth-integrated formulation. We find that urban runoff and resuspension of contaminated wetland sediments are responsible for surface water concentrations of FIB in the wetland. Model predictions show that urban runoff controls surface water concentrations at inland sites and sediment resuspension controls surface water concentrations near the mouth. Direct wash-off of bird feces into the surface water is not a significant contributor, although bird feces can contribute to the sediment bacteria load. The key parameters needed to accurately predict FIB concentrations, using a validated hydrodynamic model, are: the load due to urban runoff, sediment erodibility parameters, and sediment concentrations and surface water die-off rates of enteric bacteria. In the present study, literature values for sediment erodibility and water column die-off rates are used and average concentrations of FIB are predicted within 1/2 log unit of measurements. Total coliform are predicted more accurately than E. coli or enterococci, both in terms of magnitude and tidal variability. Since wetland-dependent animals are natural sources of FIB, and FIB survive for long periods of time and may multiply in wetland sediments, these results highlight limitations of FIB as indicators of human fecal pollution in and near

  2. Persistence of host-associated Bacteroidales gene markers and their quantitative detection in an urban and agricultural mixed prairie watershed.

    PubMed

    Tambalo, Dinah D; Fremaux, Bastien; Boa, Tyler; Yost, Christopher K

    2012-06-01

    Microbial source tracking is an emerging tool developed to protect water sources from faecal pollution. In this study, we evaluated the suitability of real time-quantitative PCR (qPCR) Taqman assays developed for detection of host-associated Bacteroidales markers in a prairie watershed. The qPCR primers and probes used in this study exhibited high accuracy (88-96% sensitivity and ≥ 99% host specificity) in detecting Bacteroidales spp. that are associated with faeces from humans, ruminants, bovines, and horses. The ruminant- and human-associated markers were also found in high concentrations within individual faecal samples, ranging from 3.4 to 7.3 log(10) marker copy numberg(-1) of individual host faeces. Following validation of host sensitivity and specificity, the host-associated Bacteroidales markers were detected in the Qu'Appelle Valley watershed of Saskatchewan, Canada which experiences a diversity of anthropogenic inputs. Concentrations of the ruminant marker were well-correlated with proximity to cattle operations and there was a correlation between the marker and Escherichia coli concentrations at these sites. Low concentrations of the human faecal marker were measured throughout the sampling sites, and may indicate a consistent influx of human faecal pollution into the watershed area. Persistence of each of the Bacteroidales host-associated marker was also studied in situ. The results indicated that the markers persist for shorter periods of time (99% decay in <8 days) compared with the conventional E. coli marker (99% decay in >15 days), suggesting they are effective at detecting recent faecal contamination events. The levels of Bacteroidales markers and E. coli counts did not correlate with the presence of the pathogenic bacteria, Salmonella spp. or Campylobacter spp. detected in the Qu'Appelle Valley. Collectively, the results obtained in this study demonstrated that the qPCR approach for detecting host-associated Bacteroidales spp. markers can be a

  3. Increases in fecal coliform bacteria resulting from centrifugal dewatering of digested biosolids.

    PubMed

    Qi, Yinan; Dentel, Steven K; Herson, Diane S

    2007-02-01

    In many countries, the classification of biosolids for disposal purposes can be based, in part, on fecal coliform levels, with alternative criteria also available based on the stabilization process used, such as anaerobic digestion. The assumption that these alternative criteria provide equivalent protection may be flawed. This paper demonstrates that fecal coliform levels determined after digestion do not always indicate the bacterial levels after the same biosolids have been dewatered by centrifugation. In samples from mesophilic digestion, half had significant increases in coliform numbers (P<0.05) with up to one order of magnitude increase during centrifugation, suggesting coliform regrowth. Thermophilically digested samples had significant increases of several orders of magnitude during dewatering, more likely from reactivation of viable but non-culturable coliforms than from regrowth. In other cases, centrifugation induced coliform regrowth or reactivation upon incubation and storage of dewatered samples, but not digested samples. These 2-3 order of magnitude increases occurred with both 25 and 37 degrees C incubations. Coliform increases continued for up to 5 days, then gradually declined. However, by day 20 coliform numbers were still 2 orders of magnitude greater than when originally sampled. The magnitude of the increases could be due either to regrowth or reactivation, but the nature of the longer-term increases--also seen in biosolids/soil mixtures--suggests regrowth. Differences in numbers between digested and dewatered samples could not be duplicated with high shear processing in lab-scale devices, with nitrogen purging to remove volatile or gaseous constituents, or with redilution using centrate. They could not be attributed to enumeration methods, to interference of Bacillus spp. on apparent coliform counts, or to temperature changes. The increases have practical implications in the use of fecal coliform or alternative criteria to define pathogen

  4. Differential decomposition of bacterial and viral fecal indicators in common human pollution types.

    PubMed

    Wanjugi, Pauline; Sivaganesan, Mano; Korajkic, Asja; Kelty, Catherine A; McMinn, Brian; Ulrich, Robert; Harwood, Valerie J; Shanks, Orin C

    2016-11-15

    Understanding the decomposition of microorganisms associated with different human fecal pollution types is necessary for proper implementation of many water quality management practices, as well as predicting associated public health risks. Here, the decomposition of select cultivated and molecular indicators of fecal pollution originating from fresh human feces, septage, and primary effluent sewage in a subtropical marine environment was assessed over a six day period with an emphasis on the influence of ambient sunlight and indigenous microbiota. Ambient water mixed with each fecal pollution type was placed in dialysis bags and incubated in situ in a submersible aquatic mesocosm. Genetic and cultivated fecal indicators including fecal indicator bacteria (enterococci, E. coli, and Bacteroidales), coliphage (somatic and F+), Bacteroides fragilis phage (GB-124), and human-associated genetic indicators (HF183/BacR287 and HumM2) were measured in each sample. Simple linear regression assessing treatment trends in each pollution type over time showed significant decay (p ≤ 0.05) in most treatments for feces and sewage (27/28 and 32/40, respectively), compared to septage (6/26). A two-way analysis of variance of log10 reduction values for sewage and feces experiments indicated that treatments differentially impact survival of cultivated bacteria, cultivated phage, and genetic indicators. Findings suggest that sunlight is critical for phage decay, and indigenous microbiota play a lesser role. For bacterial cultivated and genetic indicators, the influence of indigenous microbiota varied by pollution type. This study offers new insights on the decomposition of common human fecal pollution types in a subtropical marine environment with important implications for water quality management applications. Published by Elsevier Ltd.

  5. Physical and biological factors influencing environmental sources of fecal indicator bacteria in surface water

    USGS Publications Warehouse

    Whitman, Richard L.; Nevers, Meredith B.; Przybyla-Kelly, Katarzyna; Byappanahalli, Muruleedhara N.; Sadowsky, Michael J.; Whitman, Richard L.

    2011-01-01

    This paper describes the environmental populations of faecal indicator bacteria, and the processes by which these populations become nonpoint sources and influence nearshore water quality. The different possible sources of these indicator bacteria are presented. These include groundwater, springs and seeps, aquatic sediments, beach sand, birds, Cladophora and plant wrack. Also discussed are the environmental factors (moisture, sunlight, temperature and salinity) influencing their survival.

  6. Effect of Temperature, Grain Size and Organic Content on Persistence of Fecal Indicator Bacteria in Aquatic Sediments

    NASA Astrophysics Data System (ADS)

    Martinez, N. T.; Juhl, A. R.; O'Mullan, G. D.

    2011-12-01

    Pathogenic bacteria from poorly treated sewage present a health threat in recreational waters. Sewage derived bacteria can attach to particles and sink to the bottom, where they may persist longer than in the water column. If sewage derived bacteria persist, contaminated sediments may function as a reservoir for indicator bacteria and pathogens that can be resuspended, recontaminating the water column. We quantified the persistence of the fecal indicator bacteria (FIB), Escherichia coli and Enterococcus sp., in aquatic sediment microcosms in relation to sediment organic content, grain size and temperature. Surface sediment used for microcosms came from 5 near shore Hudson River estuary sites with different grain size distributions and organic content. Sediments from each location were divided into three separate containers that were then incubated in darkness at 18°, 25° and 30° C for several weeks. Subsamples were collected from each microcosm approximately weekly to track the decay of the FIB as a function of time. Duration required for 90% decay in different microcosms ranged from 6.7 to 63 days for E. coli and 5.1 to 60 days for Enterococcus sp., longer than has been typically observed in the water column. Our measurements of E. coli persistence were also longer than described in previous work. We found that E. coli persistence was strongly related to temperature with faster decay at higher temperatures. In contrast, Enterococcus sp. persistence was weakly related to temperature but was strongly related to sediment organic content and grain size distribution with decay rate increasing in sediments of low organic content and coarser grain size. Quantification of FIB persistence in sediment reservoirs can be used in water quality and public health predictions. The contrasting responses of E. coli and Enterococcus sp. persistence to sediment characteristics implies different suitability as indicators depending on environmental conditions.

  7. Characterizing relationships among fecal indicator bacteria, microbial source tracking markers, and associated waterborne pathogen occurrence in stream water and sediments in a mixed land use watershed

    USDA-ARS?s Scientific Manuscript database

    Bed sediments of streams and rivers may store high concentrations of fecal indicator bacteria (FIB) and pathogens. These contaminants can be mobilized into the water column due to resuspension events, thus affecting overall water quality. Along with the contaminants, other markers such as microbia...

  8. Evaluation of a real-time quantitative PCR method with propidium monazide treatment for analyses of viable fecal indicator bacteria in wastewater samples

    EPA Science Inventory

    The U.S. EPA is currently evaluating rapid, real-time quantitative PCR (qPCR) methods for determining recreational water quality based on measurements of fecal indicator bacteria DNA sequences. In order to potentially use qPCR for other Clean Water Act needs, such as updating cri...

  9. Variations in the fecal occurrences of antimicrobial-resistant bacteria are greater between seasons than between "raised without antibiotics" and "conventional" cattle production systems

    USDA-ARS?s Scientific Manuscript database

    The objective of this study was to compare, over a year, fecal occurrences of antimicrobial-resistant bacteria (ARB) in fed-cattle between two production systems: "raised without antibiotics" (RWA) and “conventional” (CONV). Feces were recovered from colons at a commercial beef processing plant obta...

  10. Rainfall and tillage effects on transport of fecal bacteria and sex hormones 17ß-estradiol and testosterone from broiler litter applications to a Georgia Piedmont Ultisol

    USDA-ARS?s Scientific Manuscript database

    Poultry litter provides plant nutrients for crop and pasture production. Fecal bacteria, sex hormones (17ß-estradiol and testosterone) and antibiotic residues are litter components, however, that may contaminate surface waters and become a public health risk. Our objective was to quantify transpor...

  11. Characterizing relationships among fecal indicator bacteria, microbial source tracking markers, and associated waterborne pathogen occurrence in stream water and sediments in a mixed land use watershed

    EPA Science Inventory

    Bed sediments of streams and rivers may store high concentrations of fecal indicator bacteria (FIB) and pathogens. Due to resuspension events, these contaminants can be mobilized into the water column and affect overall water quality. Other bacterial indicators such as microbial ...

  12. Evaluation of a real-time quantitative PCR method with propidium monazide treatment for analyses of viable fecal indicator bacteria in wastewater samples

    EPA Science Inventory

    The U.S. EPA is currently evaluating rapid, real-time quantitative PCR (qPCR) methods for determining recreational water quality based on measurements of fecal indicator bacteria DNA sequences. In order to potentially use qPCR for other Clean Water Act needs, such as updating cri...

  13. Influences of sample interference and interference controls on quantification of enterococci fecal indicator bacteria in surface water samples by the qPCR method

    EPA Science Inventory

    A quantitative polymerase chain reaction (qPCR) method for the detection of entercocci fecal indicator bacteria has been shown to be generally applicable for the analysis of temperate fresh (Great Lakes) and marine coastal waters and for providing risk-based determinations of wat...

  14. Storm flow dynamics and loads of fecal bacteria associated with ponds in southern piedmont and coastal plain watersheds with animal agriculture

    USDA-ARS?s Scientific Manuscript database

    Storm events that increase hydrologic flow rates can disturb sediments and produce overland runoff in watersheds with animal agriculture, and, thus, can increase surface water concentrations of fecal bacteria and risk to public health. We tested the hypothesis that strategically placed ponds in wate...

  15. Characterizing relationships among fecal indicator bacteria, microbial source tracking markers, and associated waterborne pathogen occurrence in stream water and sediments in a mixed land use watershed

    EPA Science Inventory

    Bed sediments of streams and rivers may store high concentrations of fecal indicator bacteria (FIB) and pathogens. Due to resuspension events, these contaminants can be mobilized into the water column and affect overall water quality. Other bacterial indicators such as microbial ...

  16. Differential Decay of Cattle-associated Fecal Indicator Bacteria and Microbial Source Tracking Markers in Fresh and Marine Water (ASM 2017 Presentation)

    EPA Science Inventory

    Background: Fecal indicator bacteria (FIB) have a long history of use in the assessment of the microbial quality of recreational waters. However, quantification of FIB provides no information about the pollution source(s) and relatively little is known about their fate in the amb...

  17. Influences of sample interference and interference controls on quantification of enterococci fecal indicator bacteria in surface water samples by the qPCR method

    EPA Science Inventory

    A quantitative polymerase chain reaction (qPCR) method for the detection of entercocci fecal indicator bacteria has been shown to be generally applicable for the analysis of temperate fresh (Great Lakes) and marine coastal waters and for providing risk-based determinations of wat...

  18. TEMPORAL AND SPATIAL VARIABILITY OF FECAL INDICATOR BACTERIA: IMPLICATIONS FOR THE APPLICATION OF MST METHODOLOGIES TO DIFFERENTIATE SOURCES OF FECAL CONTAMINATION

    EPA Science Inventory

    Temporal variability in the gastrointestinal flora of animals impacting water resources with fecal material can be one of the factors producing low source identification rates when applying microbial source tracking (MST) methods. Understanding how bacterial species and genotype...

  19. TEMPORAL AND SPATIAL VARIABILITY OF FECAL INDICATOR BACTERIA: IMPLICATIONS FOR THE APPLICATION OF MST METHODOLOGIES TO DIFFERENTIATE SOURCES OF FECAL CONTAMINATION

    EPA Science Inventory

    Temporal variability in the gastrointestinal flora of animals impacting water resources with fecal material can be one of the factors producing low source identification rates when applying microbial source tracking (MST) methods. Understanding how bacterial species and genotype...

  20. Unsealed Tubewells Lead to Increased Fecal Contamination of Drinking Water

    PubMed Central

    Knappett, Peter S. K.; McKay, Larry D.; Layton, Alice; Williams, Daniel E.; Alam, Md. J.; Mailloux, Brian J.; Ferguson, Andrew S.; Culligan, Patricia J.; Serre, Marc L.; Emch, Michael; Ahmed, Kazi M.; Sayler, Gary S.; van Geen, Alexander

    2013-01-01

    Bangladesh is underlain by shallow aquifers in which millions of drinking water wells are emplaced without annular seals. Fecal contamination has been widely detected in private tubewells. To evaluate the impact of well construction on microbial water quality 35 private tubewells (11 with intact cement platforms, 19 without) and 17 monitoring wells (11 with the annulus sealed with cement, 6 unsealed) were monitored for cultured E. coli over 18 months. Additionally, two “snap shot” sampling events were performed on a subset of wells during late-dry and early-wet seasons, wherein the fecal indicator bacteria (FIB) E. coli, Bacteroidales and the pathogenicity genes eltA (ETEC E. coli), ipaH (Shigella) and 40/41 hexon (adenovirus) were detected using qPCR. No difference in E. coli detection frequency was found between tubewells with and without platforms. Unsealed private wells, however, contained cultured E. coli more frequently and higher concentrations of FIB than sealed monitoring wells (p<0.05), suggestive of rapid downward flow along unsealed annuli. As a group the pathogens ETEC, Shigella and adenovirus were detected more frequently (10/22) during the wet season than the dry season (2/20). This suggests proper sealing of private tubewell annuli may lead to substantial improvements in microbial drinking water quality. PMID:23165714

  1. Unsealed tubewells lead to increased fecal contamination of drinking water.

    PubMed

    Knappett, Peter S K; McKay, Larry D; Layton, Alice; Williams, Daniel E; Alam, Md J; Mailloux, Brian J; Ferguson, Andrew S; Culligan, Patricia J; Serre, Marc L; Emch, Michael; Ahmed, Kazi M; Sayler, Gary S; van Geen, Alexander

    2012-12-01

    Bangladesh is underlain by shallow aquifers in which millions of drinking water wells are emplaced without annular seals. Fecal contamination has been widely detected in private tubewells. To evaluate the impact of well construction on microbial water quality 35 private tubewells (11 with intact cement platforms, 19 without) and 17 monitoring wells (11 with the annulus sealed with cement, six unsealed) were monitored for culturable Escherichia coli over 18 months. Additionally, two 'snapshot' sampling events were performed on a subset of wells during late-dry and early-wet seasons, wherein the fecal indicator bacteria (FIB) E. coli, Bacteroidales and the pathogenicity genes eltA (enterotoxigenic E. coli; ETEC), ipaH (Shigella) and 40/41 hexon (adenovirus) were detected using quantitative polymerase chain reaction (qPCR). No difference in E. coli detection frequency was found between tubewells with and without platforms. Unsealed private wells, however, contained culturable E. coli more frequently and higher concentrations of FIB than sealed monitoring wells (p < 0.05), suggestive of rapid downward flow along unsealed annuli. As a group the pathogens ETEC, Shigella and adenovirus were detected more frequently (10/22) during the wet season than the dry season (2/20). This suggests proper sealing of private tubewell annuli may lead to substantial improvements in microbial drinking water quality.

  2. The Seasonality of Fecal Coliform Bacteria Pollution and its Influence on Closures of Shellfish Harvesting Areas in Mississippi Sound

    PubMed Central

    Chigbu, Paulinus; Gordon, Scott; Tchounwou, Paul B.

    2005-01-01

    Runoff from agricultural lands and farm animal feedlots is one of the major sources of fecal coliforms in surface waters, and fecal coliform (FC) bacteria concentrations tend to vary with season because of seasonal variations in climatic factors. However, El Niño -Southern Oscillation (ENSO) events may affect the extent and patterns of seasonality in FC levels in coastal waters. Water quality monitoring data for shellfish management collected during El Niño (1990, 1992, 1997), and La Niña (1999, 2000) years were analyzed to evaluate the extent to which these events influenced Pearl River stage, and bacterial levels, water temperature, and salinity in the western part of Mississippi Sound. Models to predict FC levels in relation to various environmental factors were also developed. In 1990, 1992 and 1997, FC geometric mean counts peaked in late winter (January/February) reaching 120 MPN (February 1990), 165 MPN (January 1992), and 86 MPN (January 1997), and then decreased considerably during spring and summer (1.2 – 19 MPN). Thereafter, FC abundance increased slightly in fall and early winter (1.9 – 24 MPN). Fecal coliform abundance during the 2000 La Niña year was much lower (1.0 – 10.3 MPN) than in 1992 (1.2 – 165 MPN), and showed no seasonal pattern from January to August, perhaps due to the relative scarcity of rainfall in 2000. In 1995 (ENSO neutral year), peak geometric mean FC count (46 MPN) was lower than during El Niño years and occurred in early spring (March). The seasonal and between year variations in FC levels determined the number of days during which the conditionally approved shellfish growing area was opened for harvesting shellfish. For example, from January to April 1997, the area was not opened for shellfish harvesting, whereas in 2000, the number of days during which the area was opened ranged from 6 – 27 (January to April) to 24 – 26 (October to December). ENSO events thus influenced the extent and timing of the peak levels of

  3. Flow cytometric sorting of fecal bacteria after in situ hybridization with polynucleotide probes.

    PubMed

    Bruder, Lena M; Dörkes, Marcel; Fuchs, Bernhard M; Ludwig, Wolfgang; Liebl, Wolfgang

    2016-10-01

    The gut microbiome represents a key contributor to human physiology, metabolism, immune function, and nutrition. Elucidating the composition and genetics of the gut microbiota under various conditions is essential to understand how microbes function individually and as a community. Metagenomic analyses are increasingly used to study intestinal microbiota. However, for certain scientific questions it is sufficient to examine taxon-specific submetagenomes, covering selected bacterial genera in a targeted manner. Here we established a new variant of fluorescence in situ hybridization (FISH) combined with fluorescence-activated cell sorting (FACS), providing access to the genomes of specific taxa belonging to the complex community of the intestinal microbiota. In contrast to standard oligonucleotide probes, the RNA polynucleotide probe used here, which targets domain III of the 23S rRNA gene, extends the resolution power in environmental samples by increasing signal intensity. Furthermore, cells hybridized with the polynucleotide probe are not subjected to harsh pretreatments, and their genetic information remains intact. The protocol described here was tested on genus-specifically labeled cells in various samples, including complex fecal samples from different laboratory mouse types that harbor diverse intestinal microbiota. Specifically, as an example for the protocol described here, RNA polynucleotide probes could be used to label Enterococcus cells for subsequent sorting by flow cytometry. To detect and quantify enterococci in fecal samples prior to enrichment, taxon-specific PCR and qPCR detection systems have been developed. The accessibility of the genomes from taxon-specifically sorted cells for subsequent molecular analyses was demonstrated by amplification of functional genes. Copyright © 2016 Elsevier GmbH. All rights reserved.

  4. Hydrologic transport of fecal bacteria attenuated by flue gas desulfurization gypsum.

    PubMed

    Jenkins, M B; Schomberg, H H; Endale, D M; Franklin, D H; Fisher, D S

    2014-01-01

    Flue gas desulfurization (FGD) gypsum is a byproduct of coal-fired power plants. Its application to agricultural fields may increase water infiltration, reduce soil erosion, and decrease nutrient losses from applications of animal manures. It may also reduce fecal bacterial contamination of surface waters. We tested the hypothesis that FGD gypsum applications would decrease the load of and the fecal indicator bacterium from poultry litter applications. Two rainfall simulation experiments were undertaken: one in spring 2009 and one in spring 2011. Six treatments consisted of four rates of FGD gypsum (0, 2.2, 4.5, and 9.0 Mg ha) with poultry litter (13.5 Mg ha and two controls) in a randomized, complete-block design with three replications. Each replicate 4- × 6-m plot contained a single 1- × 2-m subplot that was delineated by metal plates and a flume that captured total overland flow or runoff. Rainfall was applied at ∼64 mm h. Volume of overland runoff was measured and subsampled for analysis every 10 min for 1 h. Flow-weighted concentrations, total loads, and soil concentrations of were determined. was not detected in runoff. No significant differences between treatments were observed for the 2009 rainfall simulation. However, after 3 yr of FGD gypsum applications, the highest rate of FGD gypsum resulted in decreased flow-weighted concentrations and total loads of . Flue gas desulfurization gypsum applications may be a management practice that reduces microbial contamination of surface waters from manure applied to agricultural fields in the southeastern United States. Copyright © by the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America, Inc.

  5. Bacteria holding times for fecal coliform by mFC agar method and total coliform and Escherichia coli by Colilert-18 Quanti-Tray method.

    PubMed

    Aulenbach, Brent T

    2010-02-01

    Bacteria holding-time experiments of up to 62 h were performed on five surface-water samples from four urban stream sites in the vicinity of Atlanta, GA, USA that had relatively high densities of coliform bacteria (Escherichia coli densities were all well above the US Environmental Protection Agency criterion of 126 colonies (100 ml)(-1) for recreational waters). Holding-time experiments were done for fecal coliform using the membrane filtration modified fecal coliform (mFC) agar method and for total coliform and E. coli using the Colilert-18 Quanti-Tray method. The precisions of these analytical methods were quantified. Precisions determined for fecal coliform indicated that the upper bound of the ideal range of counts could reasonably be extended upward and would improve precision. For the Colilert-18 method, analytical precisions were similar to the theoretical precisions for this method. Fecal and total coliform densities did not change significantly with holding times up to about 27 h. Limited information indicated that fecal coliform densities might be stable for holding times of up to 62 h, whereas total coliform densities might not be stable for holding times greater than about 27 h. E. coli densities were stable for holding times of up to 18 h--a shorter period than indicated from a previous studies. These results should be applicable to non-regulatory monitoring sampling designs for similar urban surface-water sample types.

  6. Cattle-level risk factors associated with fecal shedding of Shiga toxin-encoding bacteria on dairy farms, Minnesota, USA.

    PubMed

    Cho, Seongbeom; Fossler, Charles P; Diez-Gonzalez, Francisco; Wells, Scott J; Hedberg, Craig W; Kaneene, John B; Ruegg, Pamela L; Warnick, Lorin D; Bender, Jeffrey B

    2009-04-01

    The objective of this study was to identify individual cattle-level risk factors associated with fecal shedding of Shiga toxin-encoding bacteria (STB), a surrogate for Shiga toxin-producing E. coli (STEC), on 28 organic and conventional dairy farms. It was found that small organic herds (fewer than 100 cows) were associated with higher odds of Shiga toxin-encoding bacteria (STB) shedding from 2 (all cattle and all cows) of 3 cattle models, followed by small conventional herds, compared with large conventional herds. Preweaned calves [odds ratio (OR) = 2.6, 95% confidence interval (CI): 1.2, 5.7] had higher odds of shedding STB compared with adult cows. Calves more than 28 days of age (OR = 2.0, 95%CI: 1.0, 4.4) were more likely to shed STB than calves less than 28 days of age. This information may be helpful for identifying potential control strategies such as targeted vaccination or management practices.

  7. Microbial Communities and Fecal Indicator Bacteria Associated with Cladophora Mats on Beach Sites along Lake Michigan Shores

    PubMed Central

    Olapade, Ola A.; Depas, Morgan M.; Jensen, Erika T.; McLellan, Sandra L.

    2006-01-01

    A high biomasses of Cladophora, a filamentous green alga, is found mainly during the summer along the shores of Lake Michigan. In this study, the abundance and persistence of the fecal indicator bacterium Escherichia coli and sulfate-reducing bacteria (SRB) on Cladophora mats collected at Lake Michigan beaches were evaluated using both culture-based and molecular analyses. Additionally, 16S rRNA gene cloning and sequencing were used to examine the bacterial community composition. Overall, E. coli was detected in all 63 samples obtained from 11 sites, and the average levels at most beaches ranged from 2,700 CFU/100 g (wet weight) of Cladophora to 7,500 CFU/100 g of Cladophora. However, three beaches were found to have site average E. coli densities of 12,800, 21,130, and 27,950 CFU/100 g of Cladophora. The E. coli levels in the lake water collected at the same time from these three sites were less than the recommended U.S. Environmental Protection Agency limit, 235 CFU/100 ml. E. coli also persisted on Cladophora mats in microcosms at room temperature for more than 7 days, and in some experiments it persisted for as long as 28 days. The SRB densities on Cladophora mats were relatively high, ranging from 4.4 × 106 cells/g (6.64 log CFU/g) to 5.73 × 106 cells/g (6.76 log CFU/g) and accounting for between 20% and 27% of the total bacterial counts. Partial sequences of the 16S rRNA gene clones revealed a phylogenetically diverse community, in which the Cytophaga-Flavobacterium-Bacteroides cluster and the low-G+C-content gram-positive bacteria were the dominant organisms, accounting for 40% and 12.8%, respectively, of the total clone library. These results further reveal the potential public health and ecological significance of Cladophora mats that are commonly found along the shoreline of Lake Michigan, especially with regard to the potential to harbor microorganisms associated with fecal pollution and odor-causing bacteria. PMID:16517640

  8. LA35 Poultry Fecal Marker Persistence Is Correlated with That of Indicators and Pathogens in Environmental Waters

    PubMed Central

    Nayak, Bina; Weidhaas, Jennifer

    2015-01-01

    Disposal of fecally contaminated poultry litter by land application can deliver pathogens and fecal indicator bacteria (FIB) into receiving waters via runoff. While water quality is regulated by FIB enumeration, FIB testing provides inadequate information about contamination source and health risk. This microbial source tracking (MST) study compared the persistence of the Brevibacterium sp. strain LA35 16S rRNA gene (marker) for poultry litter with that of pathogens and FIB under outdoor, environmentally relevant conditions in freshwater, marine water, and sediments over 7 days. Salmonella enterica, Campylobacter jejuni, Campylobacter coli, Bacteroidales, and LA35 were enumerated by quantitative PCR (qPCR), and Enterococcus spp. and E. coli were quantified by culture and qPCR. Unlike the other bacteria, C. jejuni was not detectable after 48 h. Bacterial levels in the water column consistently declined over time and were highly correlated among species. Survival in sediments ranged from a slow decrease over time to growth, particularly in marine microcosms and for Bacteroidales. S. enterica also grew in marine sediments. Linear decay rates in water (k) ranged from −0.17 day−1 for LA35 to −3.12 day−1 for C. coli. LA35 levels correlated well with those of other bacteria in the water column but not in sediments. These observations suggest that, particularly in the water column, the fate of LA35 in aquatic environments is similar to that of FIB, C. coli, and Salmonella, supporting the hypothesis that the LA35 marker gene can be a useful tool for evaluating the impact of poultry litter on water quality and human health risk. PMID:25934617

  9. Effects of inulin chain length on fermentation by equine fecal bacteria and Streptococcus bovis

    USDA-ARS?s Scientific Manuscript database

    Fructans from pasture can be fermented by Gram-positive bacteria (e.g., Streptococcus bovis) in the equine hindgut, increasing production of lactic acid and decreasing pH. The degree of polymerization (DP) of fructans has been suggested to influence fermentation rates. The objective of the current ...

  10. Monitoring and predicting the fecal indicator bacteria concentrations from agricultural, mixed land use and urban stormwater runoff.

    PubMed

    Paule-Mercado, M A; Ventura, J S; Memon, S A; Jahng, D; Kang, J-H; Lee, C-H

    2016-04-15

    While the urban runoff are increasingly being studied as a source of fecal indicator bacteria (FIB), less is known about the occurrence of FIB in watershed with mixed land use and ongoing land use and land cover (LULC) change. In this study, Escherichia coli (EC) and fecal streptococcus (FS) were monitored from 2012 to 2013 in agricultural, mixed and urban LULC and analyzed according to the most probable number (MPN). Pearson correlation was used to determine the relationship between FIB and environmental parameters (physicochemical and hydrometeorological). Multiple linear regressions (MLR) were used to identify the significant parameters that affect the FIB concentrations and to predict the response of FIB in LULC change. Overall, the FIB concentrations were higher in urban LULC (EC=3.33-7.39; FS=3.30-7.36log10MPN/100mL) possibly because of runoff from commercial market and 100% impervious cover (IC). Also, during early-summer season; this reflects a greater persistence and growth rate of FIB in a warmer environment. During intra-event, however, the FIB concentrations varied according to site condition. Anthropogenic activities and IC influenced the correlation between the FIB concentrations and environmental parameters. Stormwater temperature (TEMP), turbidity, and TSS positively correlated with the FIB concentrations (p>0.01), since IC increased, implying an accumulation of bacterial sources in urban activities. TEMP, BOD5, turbidity, TSS, and antecedent dry days (ADD) were the most significant explanatory variables for FIB as determined in MLR, possibly because they promoted the FIB growth and survival. The model confirmed the FIB concentrations: EC (R(2)=0.71-0.85; NSE=0.72-0.86) and FS (R(2)=0.65-0.83; NSE=0.66-0.84) are predicted to increase due to urbanization. Therefore, these findings will help in stormwater monitoring strategies, designing the best management practice for FIB removal and as input data for stormwater models.

  11. Factors Influencing the Accumulation and Subsurface Transport of Fecal Indicator Bacteria near the Shoreline at Freshwater Beaches

    NASA Astrophysics Data System (ADS)

    Wu, M. Z.; O'Carroll, D. M.; Vogel, L. J.; Robinson, C. E.

    2015-12-01

    Beach sand near the shoreline acts as a reservoir for fecal contaminants with fecal indicator bacteria (FIB) often orders of magnitude higher than in adjacent surface waters. This reservoir poses a human health risk and can also act as an important non-point contamination source for surface waters. Beach water quality advisories or closures can be issued when FIB (Escherichia coli (E. coli), enterococci (ENT)) concentrations are elevated in the surface water. The factors controlling the transport and accumulation of FIB in the foreshore sand are not well understood, though this is required to manage and mitigate this source. Multiple sources may contribute to the accumulation of FIB in sand, with recent studies suggesting that the continuous influx of surface water across the sediment-water interface may be a dominant source at many beaches.The study objective was to develop understanding of the physical processes controlling the accumulation and transport of FIB in beach sand. Field measurements were combined with numerical modelling to evaluate the role of low-energy lapping waves in delivering FIB to the saturated foreshore sand at freshwater beaches. E. coli and ENT were measured at two beaches in Ontario, Canada at depths of up to 1 and 2 m, respectively, below the water table. A numerical model simulating wave-induced groundwater recirculations coupled with microbial transport (using colloid filtration theory) showed that the different FIB distributions measured at the two beaches was due mainly to the different beach slope and terrestrial groundwater flow. The model was applied to assess the impact of beach, wave and bacterial parameters on FIB accumulation. The infiltration zone width, average infiltration velocity and infiltration rate were shown to ultimately control the amount and spatial distribution of FIB in the sand. The study findings are important in understanding factors controlling the transport of FIB at the sediment-water interface of

  12. Comparing laboratory column test treatments with field profiles of fecal indicator bacteria and virus from concentrated source areas

    NASA Astrophysics Data System (ADS)

    Feighery, J.; Culligan, P.; Ferguson, A. S.; Mailloux, B. J.; McKay, L. D.; Ahmed, K.; Alam, M.; Huq, M.; Emch, M.; Serre, M. L.; Yunus, M.; van Geen, A.

    2010-12-01

    Fecal contamination of potable water supplies is prevalent throughout the developing world. In rural Bangladesh, groundwater contamination of shallow unconfined aquifers is attributed to the infiltration of fecal organisms from sewage ponds, sewage ditches and latrines. However, few studies conclusively link sources to wells at the scale required for microbial transport to occur. We present a combined field and laboratory investigation into the transport of the fecal indicator bacteria Escherichia coli (E. coli) and enteric viral indicator F+ RNA coliphage (MS-2) using drive point piezometers and extracted sediment cores. Fieldwork and coring took place in the Matlab Upazila, Bangladesh. Field measurements at the 100-cm scale were made using an array of three drive-point piezometers under highly contaminated ponds and canals over a 10-day period during the peak of the monsoon season. The profiles of E. coli detected under ponds and canals by a culture-based most probable number method were consistent with a first order filtration rate over the distances studied and filtration rates ranged from 1 - 8 m-1. In order to determine possible attachment mechanisms and the influence of sediment treatments applied in laboratory testing, duplicate column transport studies at the 10-cm scale were performed on intact cores processed immediately on-site, intact cores preserved by freezing, dried repacked sediment, acid-washed repacked sediment, and a uniform silica sand. Two ionic strengths (3.5 and 20 mM) were used to encompass the range of electrical conductivity typically found in the shallow portion of the aquifer. Columns were dissected and the attached E. coli quantified by section. Even at the solution chemistry less favorable for particle attachment (low ionic strength), filtration rates for the core tested on-site predict a transport distance of 0.5m for a 4-log unit reduction in E. coli concentration. Although the filtration rates found in the field study are lower

  13. Rainfall and tillage effects on transport of fecal bacteria and sex hormones 17beta-estradiol and testosterone from broiler litter applications to a Georgia Piedmont Ultisol.

    PubMed

    Jenkins, Michael B; Truman, Clint C; Siragusa, Gregory; Line, Eric; Bailey, J Stan; Frye, Jonathan; Endale, Dinku M; Franklin, Dorcas H; Schomberg, Harry H; Fisher, Dwight S; Sharpe, Ronald R

    2008-09-15

    Poultry litter provides nutrients for crop and pasture production; however, it also contains fecal bacteria, sex hormones (17beta-estradiol and testosterone) and antibiotic residues that may contaminate surface waters. Our objective was to quantify transport of fecal bacteria, estradiol, testosterone and antibiotic residues from a Cecil sandy loam managed since 1991 under no-till (NT) and conventional tillage (CT) to which either poultry litter (PL) or conventional fertilizer (CF) was applied based on the nitrogen needs of corn (Zea mays L) in the Southern Piedmont of NE Georgia. Simulated rainfall was applied for 60 min to 2 by 3-m field plots at a constant rate in 2004 and variable rate in 2005. Runoff was continuously measured and subsamples taken for determining flow-weighted concentrations of fecal bacteria, hormones, and antibiotic residues. Neither Salmonella, nor Campylobacter, nor antimicrobial residues were detected in litter, soil, or runoff. Differences in soil concentrations of fecal bacteria before and after rainfall simulations were observed only for Escherichia coli in the constant rainfall intensity experiment. Differences in flow-weighted concentrations were observed only for testosterone in both constant and variable intensity rainfall experiments, and were greatest for treatments that received poultry litter. Total loads of E. coli and fecal enterococci, were largest for both tillage treatments receiving poultry litter for the variable rainfall intensity. Load of testosterone was greatest for no-till plots receiving poultry litter under variable rainfall intensity. Poultry litter application rates commensurate for corn appeared to enhance only soil concentrations of E. coli, and runoff concentrations of testosterone above background levels.

  14. Fecal-indicator bacteria in the Yakima River Basin, Washington-An examination of 1999 and 2000 synoptic-sampling data and their relation to historical data

    USGS Publications Warehouse

    Morace, Jennifer L.; McKenzie, Stuart W.

    2002-11-27

    Looking forward relative to future monitoring goals, research needs, and best management practice development, four hypotheses that deal with processes and sources of bacteria were identified: (1) overland runoff transports bacteria from land surfaces to streams, (2) bacteria in the water column tend to associate with suspended matter, (3) with increasing densities of warm-blooded animals, the likelihood of fecal-coliform contamination in streams also increases, and (4) identifi- cation of bacterial sources is difficult, but must be attempted for remediation to be possible. 

  15. Herd-level risk factors associated with fecal shedding of Shiga toxin-encoding bacteria on dairy farms in Minnesota, USA.

    PubMed

    Cho, Seongbeom; Fossler, Charles P; Diez-Gonzalez, Francisco; Wells, Scott J; Hedberg, Craig W; Kaneene, John B; Ruegg, Pamela L; Warnick, Lorin D; Bender, Jeffrey B

    2013-07-01

    This study aimed to identify herd-level risk factors associated with fecal shedding of Shiga toxin-encoding bacteria (STB) on dairy cattle farms in Minnesota, USA. After adjustment for farm size, risk factors included: use of total mixed ration (TMR) for lactating dairy cows [odds ratio (OR) = 3.0; 95% confidence interval (CI): 1.8 to 5.1], no use of monensin for weaned calves (OR = 4.8, 95% CI: 2.5, 9.3), and no use of decoquinate for preweaned calves (OR = 2.2, 95% CI: 1.4, 3.6). Fecal shedding of STB was more common in small herds (< 100 cows, OR = 3.6, 95% CI: 2.1, 6.2) than in large herds (≥ 100 cows). Herd management factors related to cattle feeding practices were associated with fecal shedding of STB.

  16. Integrated modeling approach using SELECT and SWAT models to simulate source loading and in-stream conditions of fecal indicator bacteria.

    NASA Astrophysics Data System (ADS)

    Ranatunga, T.

    2016-12-01

    Modeling of fate and transport of fecal bacteria in a watershed is generally a processed based approach that considers releases from manure, point sources, and septic systems. Overland transport with water and sediments, infiltration into soils, transport in the vadose zone and groundwater, die-off and growth processes, and in-stream transport are considered as the other major processes in bacteria simulation. This presentation will discuss a simulation of fecal indicator bacteria (E.coli) source loading and in-stream conditions of a non-tidal watershed (Cedar Bayou Watershed) in South Central Texas using two models; Spatially Explicit Load Enrichment Calculation Tool (SELECT) and Soil and Water Assessment Tool (SWAT). Furthermore, it will discuss a probable approach of bacteria source load reduction in order to meet the water quality standards in the streams. The selected watershed is listed as having levels of fecal indicator bacteria that posed a risk for contact recreation and wading by the Texas Commission of Environmental Quality (TCEQ). The SELECT modeling approach was used in estimating the bacteria source loading from land categories. Major bacteria sources considered were, failing septic systems, discharges from wastewater treatment facilities, excreta from livestock (Cattle, Horses, Sheep and Goat), excreta from Wildlife (Feral Hogs, and Deer), Pet waste (mainly from Dogs), and runoff from urban surfaces. The estimated source loads were input to the SWAT model in order to simulate the transport through the land and in-stream conditions. The calibrated SWAT model was then used to estimate the indicator bacteria in-stream concentrations for future years based on H-GAC's regional land use, population and household projections (up to 2040). Based on the in-stream reductions required to meet the water quality standards, the corresponding required source load reductions were estimated.

  17. Survival, transport, and sources of fecal bacteria in streams and survival in land-applied poultry litter in the upper Shoal Creek basin, southwestern Missouri, 2001-2002

    USGS Publications Warehouse

    Schumacher, John G.

    2003-01-01

    Densities of fecal coliform bacteria along a 5.7-mi (mile) reach of Shoal Creek extending upstream from State Highway 97 (site 3) to State Highway W (site 2) and in two tributaries along this reach exceeded the Missouri Department of Natural Resources (MDNR) standard of 200 col/100 mL (colonies per 100 milliliters) for whole-body contact recreation. A combination of techniques was used in this report to provide information on the source, transport, and survival of fecal bacteria along this reach of Shoal Creek. Results of water-quality samples collected during dye-trace and seepage studies indicated that at summer low base-flow conditions, pastured cattle likely were a substantial source of fecal bacteria in Shoal Creek at the MDNR monitoring site (site 3) at State Highway 97. Using repeat element Polymerase Chain Reaction (rep-PCR), cattle were the presumptive source of about 50 percent of the Escherichia coli (E. coli) isolates in water samples from site 3. Cattle, horses, and humans were the most common presumptive source of E. coli isolates at sites further upstream. Poultry was identified by rep-PCR as a major source of E. coli in Pogue Creek, a tributary in the upper part of the study area. Results of the rep-PCR were in general agreement with the detection and distribution of trace concentrations of organic compounds commonly associated with human wastewater, such as caffeine, the antimicrobial agent triclosan, and the pharmaceutical compounds acetaminophen and thiabendazole (a common cattle anthelmintic). Significant inputs of fecal bacteria to Shoal Creek occurred along a 1.6-mi reach of Shoal Creek immediately upstream from site 3. During a 36-hour period in July 2001, average densities of fecal coliform and E. coli bacteria increased from less than or equal to 500 col/100 mL upstream from this stream reach (sample site 2c) to 2,100 and 1,400 col/100 mL, respectively, at the MDNR sampling site. Fecal bacteria densities exhibited diurnal variability at all

  18. Nutrient and Fecal Indicator Bacteria Inputs from Submarine Groundwater Discharge on the North Shore of Kauai

    NASA Astrophysics Data System (ADS)

    Knee, K.; Boehm, A.; Paytan, A.

    2006-12-01

    Submarine groundwater discharge (SGD) is a potentially important source of freshwater, nutrients, and pollution to the coastal ocean, yet it has not been well quantified for most locations. This poster investigates the impacts of SGD in Hanalei Bay, Ha'ena, and Princeville, three locations on the north shore of Kaua'i, Hawai'i, during two sampling trips conducted in March 2005 and August 2006. The short-lived isotopes of radium, 223Ra and 224Ra, were used to calculate the flux of groundwater into the coastal ocean in these areas. Nutrient concentrations and levels of total coliform bacteria, E. coli, and Enterococcus were measured in groundwater, the nearshore ocean, rivers, and streams. Fluxes of nutrients and bacteria associated with groundwater discharge were estimated. The results indicate that nutrient subsidies to the nearshore ocean from SGD may be comparable to those from rivers and streams, and that FIB subsidies may also be significant in some areas. More sampling is necessary to assess seasonal variability, which may be significant.

  19. Sunlight inactivation of fecal indicator bacteria in open-water unit process treatment wetlands: Modeling endogenous and exogenous inactivation rates.

    PubMed

    Nguyen, Mi T; Jasper, Justin T; Boehm, Alexandria B; Nelson, Kara L

    2015-10-15

    A pilot-scale open-water unit process wetland was monitored for one year and found to be effective in enhancing sunlight inactivation of fecal indicator bacteria (FIB). The removal of Escherichia coli and enterococci in the open-water wetland receiving non-disinfected secondary municipal wastewater reached 3 logs and 2 logs in summer time, respectively. Pigmented enterococci were shown to be significantly more resistant to sunlight inactivation than non-pigmented enterococci. A model was developed to predict the inactivation of E. coli, and pigmented and non-pigmented enterococci that accounts for endogenous and exogenous sunlight inactivation mechanisms and dark processes. Endogenous inactivation rates were modeled using the sum of UVA and UVB irradiance. Exogenous inactivation was only significant for enterococci, and was modeled as a function of steady-state singlet oxygen concentration. The rate constants were determined from lab experiments and an empirical correction factor was used to account for differences between lab and field conditions. The model was used to predict removal rate constants for FIB in the pilot-scale wetland; considering the variability of the monitoring data, there was general agreement between the modeled values and those determined from measurements. Using the model, we estimate that open-water wetlands at 40° latitude with practical sizes can achieve 3-log (99.9%) removal of E. coli and non-pigmented enterococci throughout the year [5.5 ha and 7.0 ha per million gallons of wastewater effluent per day (MGD), respectively]. Differences in sunlight inactivation rates observed between pigmented and non-pigmented enterococci, as well as between lab-cultured and indigenous wastewater bacteria highlight the challenges of using FIB as model organisms for actual pathogens in natural sunlit environments.

  20. Evaluation of the quality of coastal bathing waters in Spain through fecal bacteria Escherichia coli and Enterococcus.

    PubMed

    Aragonés, L; López, I; Palazón, A; López-Úbeda, R; García, C

    2016-10-01

    Sun. and beach tourism is very important to the economy of Spain, so the control of the quality of the environment on the beaches is essential. Therefore, the analysis and control of the quality of bathing water is necessary, which is defined by the European Directive 2006/7/EC as excellent, good or sufficient depending on the presence of microbiological contamination or other organisms or waste presenting a risk to bathers' health. For that, 1392 beaches of the Iberian Peninsula and its islands were analysed, taking into account: fecal bacteria (Escherichia coli and Enterococcus), physical characteristics of sediment, level of urbanization, climatic and anthropogenic factors, and maritime climate. Thus, it was observed that urban sand beaches located in seas with fewer hours of sunshine and important tide have higher concentrations of E. coli and Enterococcus. There is also an indirect relationship between these microorganisms with salinity (R(2) 0.746 for E. coli and 0.606 for Enterococcus), temperature (R(2) 0.743 for E. coli and 0.604 for Enterococcus) and hours of sunshine (R(2) 0.781 for E. coli and 0.706 for Enterococcus), while this relationship is direct with rainfall (R(2) 0.640 for E. coli and 0.607 for Enterococcus) or wave height (R(2) 0.769 for E. coli and 0.601 for Enterococcus). From all this, it follows that the Directive 2006/7/EC should define more specific criteria as to the place and time of sampling, and take into account the different environment variables that influence the survival of bacteria, so that the results may reflect reality, and avoid staff responsible for sampling freely choose the place and time of sampling.

  1. Assessing environmental impacts of treated wastewater through monitoring of fecal indicator bacteria and salinity in irrigated soils.

    PubMed

    McLain, Jean E T; Williams, Clinton F

    2012-03-01

    To assess the potential for treated wastewater irrigation to impact levels of fecal indicator bacteria (FIB) and salinity in irrigated soils, levels of Escherichia coli, Enterococcus, and environmental covariates were measured in a treated wastewater holding pond (irrigation source water), water leaving the irrigation system, and in irrigated soils over 2 years in a municipal parkland in Arizona. Higher E. coli levels were measured in the pond in winter (56 CFU 100 mL(-1)) than in summer (17 CFU 100 mL(-1)); however, in the irrigation system, levels of FIB decreased from summer (26 CFU 100 mL(-1)) to winter (4 CFU 100 mL(-1)), possibly related to low winter water use and corresponding death of residual bacteria within the system. For over 2 years, no increase in FIB was found in irrigated soils, though highest E. coli levels (700 CFU g(-1) soil) were measured in deeper (20-25 cm) soils during summer. Measurements of water inputs vs. potential evapotranspiration indicate that irrigation levels may have been sufficient to generate bacterial percolation to deeper soil layers during summer. No overall increase in soil salinity resulting from treated wastewater irrigation was detected, but distinct seasonal peaks as high as 4 ds m(-1) occurred during both summers. The peaks significantly declined in winter when surface ET abated and more favorable water balances could be maintained. Monitoring of seasonal shifts in irrigation water quality and/or factors correlated with increases and decreases in FIB will aid in identification of any public health or environmental risks that could arise from the use of treated wastewater for irrigation.

  2. The compartment bag test (CBT) for enumerating fecal indicator bacteria: Basis for design and interpretation of results.

    PubMed

    Gronewold, Andrew D; Sobsey, Mark D; McMahan, Lanakila

    2017-06-01

    For the past several years, the compartment bag test (CBT) has been employed in water quality monitoring and public health protection around the world. To date, however, the statistical basis for the design and recommended procedures for enumerating fecal indicator bacteria (FIB) concentrations from CBT results have not been formally documented. Here, we provide that documentation following protocols for communicating the evolution of similar water quality testing procedures. We begin with an overview of the statistical theory behind the CBT, followed by a description of how that theory was applied to determine an optimal CBT design. We then provide recommendations for interpreting CBT results, including procedures for estimating quantiles of the FIB concentration probability distribution, and the confidence of compliance with recognized water quality guidelines. We synthesize these values in custom user-oriented 'look-up' tables similar to those developed for other FIB water quality testing methods. Modified versions of our tables are currently distributed commercially as part of the CBT testing kit.

  3. Sensitivity Analysis of Factors Influencing the Fate and Transport of Fecal Indicator Bacteria in Southern Lake Michigan

    NASA Astrophysics Data System (ADS)

    Thupaki, P.; Phanikumar, M. S.; Schwab, D. J.; Whitman, R. L.; Nevers, M. B.; Shively, D. A.

    2007-05-01

    To understand the factors that influence the fate and transport of fecal indicator bacteria (FIB) in the nearshore waters of the Great Lakes, we examined two southern Lake Michigan beaches (as well as the tributaries discharging into the lake in the vicinity of the beaches). A three-dimensional, σ-coordinate Princeton Ocean Model (POM) with a nested-grid was used to describe wind-driven circulation in Lake Michigan. A biological model coupled to the hydrodynamic and temperature fields in the lake was used to describe the observed FIB levels near the beaches. We report simulation results for the summers of 2004 and 2006. Inactivation of pathogens in the nearshore region is influenced by a complex set of factors including solar insolation, water temperature, settling of particulate matter, resuspension, turbulent diffusion, loading from tributaries etc. Efforts to systematically quantify the relative contributions of these complex and often inter-related processes are somewhat limited, especially for freshwater environments. Here we describe sensitivity analyses based on our numerical simulations with the objective of ranking the various processes involved in terms of their relative importance. We also examine the performance of different mathematical formulations of inactivation in order to identify their relative merits.

  4. In vitro fermentation of oat bran obtained by debranning with a mixed culture of human fecal bacteria.

    PubMed

    Kedia, Gopal; Vázquez, José A; Charalampopoulos, Dimitris; Pandiella, Severino S

    2009-04-01

    The prebiotic potential of oat samples was investigated by in vitro shaker-flask anaerobic fermentations with human fecal cultures. The oat bran fraction was obtained by debranning and was compared with other carbon sources such as whole oat flour, glucose, and fructo-oligosaccharide. The oat bran fraction showed a decrease in culturable anaerobes and clostridia and an increase in bifidobacteria and lactobacilli populations. A similar pattern was observed in fructo-oligosaccharide. Butyrate production was higher in oat bran compared to glucose and similar to that in fructo-oligosaccharide. Production of propionate was higher in the two oat media than in fructo-oligosaccharide and glucose, which can be used as energy source by the liver. This study suggests that the oat bran fraction obtained by debranning is digested by the gut ecosystem and increases the population of beneficial bacteria in the indigenous gut microbiota. This medium also provides an energy source preferred by colonocytes when it is metabolized by the gut flora.

  5. Effects of land uses on fecal indicator bacteria in the water and soil of a tropical watershed.

    PubMed

    Goto, Dustin K; Yan, Tao

    2011-01-01

    Effects of different land uses on densities of Escherichia coli, enterococci, and Clostridium perfringens in the water and soil of a tropical watershed were investigated. Densities of fecal indicator bacteria (FIBs) in the watershed exhibited a clear land-use dependency in the stream water. Significantly higher concentrations were detected in the urban portion of the stream (417, 420, and 44 CFU 100 mL(-1) for E. coli, enterococci, and C. perfringens, respectively) than in the forest portion (54, 32, and 5 CFU 100 mL(-1) for E. coli, enterococci, and C. perfringens, respectively). High concentrations of FIBs were also detected in the soil of the watershed with concentration ranges of 603-1,820,000, 69-17,000, and 0-525 CFU 100 g soil(-1) for E. coli, enterococci, and C. perfringens, respectively, which however were not affected by the different land uses. Prior cumulative rainfall significantly correlated with concentrations of E. coli and enterococci in the urban stream water (r=0.73-0.87, P<0.05), but not with the alternative FIB C. perfringens. Poor correlations were observed in the forest reach of the stream for all FIBs. Furthermore, the concentration of C. perfringens only correlated strongly and significantly with E. coli and enterococci in stream water (r=0.70-0.82, P<0.05), but not in tropical soil, indicating different survival and transport behaviors.

  6. Risk factors for infection with pathogenic and antimicrobial-resistant fecal bacteria in northern elephant seals in California.

    PubMed

    Stoddard, Robyn A; Atwill, Edward R; Gulland, Frances M D; Miller, Melissa A; Dabritz, Haydee A; Paradies, Dave M; Worcester, Karen R; Jang, Spencer; Lawrence, Judy; Byrne, Barbara A; Conrad, Patricia A

    2008-01-01

    The goal of this study was to identify potential environmental and demographic factors associated with Campylobacter jejuni (C. jejuni), Salmonella enterica (Salmonella spp.), and antimicrobial-resistant Escherichia coli (E. coli) infection in northern elephant seals stranded along the California coastline. E. coli, Salmonella spp., and C. jejuni were isolated from rectal swabs from 196 juvenile northern elephant seals, which were found stranded and alive along the California coast and brought to The Marine Mammal Center in Sausalito, California, for rehabilitation. Gender, weight, county where the animal stranded, month stranded, coastal human population density, exposure to sewage outfall or freshwater outflow (river or stream), and cumulative precipitation in the previous 24 hours, seven days, 30 days, 90 days, and 180 days were analyzed as potential risk factors for infection. The odds of C. jejuni and antimicrobial-resistant E. coli were higher in feces of seals stranded at sites with higher levels of freshwater outflow compared with lower levels of freshwater outflow. The odds of Salmonella spp. in feces were 5.4 times greater in seals stranded in locations with lower levels of 30-day cumulative precipitation, along with substantially lower odds of Salmonella shedding for seals stranded in Monterey or Santa Cruz county compared with seals stranded in regions further north or south of this central California location. Juvenile northern elephant seals that have entered the water are being colonized by antimicrobial-resistant and pathogenic fecal bacteria that may be acquired from terrestrial sources transmitted via river and surface waters.

  7. Isolation and Identification of Quercetin Degrading Bacteria from Human Fecal Microbes

    PubMed Central

    Zhang, Zhichao; Peng, Xichun; Li, Shaoting; Zhang, Ning; wang, Yong; Wei, Hua

    2014-01-01

    Quercetin has a wide range of biological properties. The gut microflora can often modulate its biological activity and their potential health effects. There still is a lack of information about gut bacteria involving in this process. The strains of gut microbes from human feces that can transform quercetin were isolated and identified by in vitro fermentation. The results showed that Escherichia coli, Stretococcus lutetiensis, Lactobacillus acidophilus, Weissella confusa, Enterococcus gilvus, Clostridium perfringens and Bacteroides fragilis have the various ability of degrading quercetin. Among them, C. perfringens and B. fragilis were discovered to have the strongest ability of degrading quercetin. Additionally, quercetin can't inhibit the growth of C. perfringens. In conclusion, many species of gut microbiota can degrade quercetin, but their ability are different. PMID:24594786

  8. Isolation and identification of quercetin degrading bacteria from human fecal microbes.

    PubMed

    Zhang, Zhichao; Peng, Xichun; Li, Shaoting; Zhang, Ning; Wang, Yong; Wei, Hua

    2014-01-01

    Quercetin has a wide range of biological properties. The gut microflora can often modulate its biological activity and their potential health effects. There still is a lack of information about gut bacteria involving in this process. The strains of gut microbes from human feces that can transform quercetin were isolated and identified by in vitro fermentation. The results showed that Escherichia coli, Stretococcus lutetiensis, Lactobacillus acidophilus, Weissella confusa, Enterococcus gilvus, Clostridium perfringens and Bacteroides fragilis have the various ability of degrading quercetin. Among them, C. perfringens and B. fragilis were discovered to have the strongest ability of degrading quercetin. Additionally, quercetin can't inhibit the growth of C. perfringens. In conclusion, many species of gut microbiota can degrade quercetin, but their ability are different.

  9. Antibiotic resistance among bacteria isolated from seawater and penguin fecal samples collected near Palmer Station, Antarctica.

    PubMed

    Miller, Robert V; Gammon, Katharine; Day, Martin J

    2009-01-01

    Antibiotic resistance in aquatic bacteria has increased steadily as a consequence of the widespread use of antibiotics, but practice and international treaty should have limited antibiotic contamination in Antarctica. We estimated antibiotic resistance in microorganisms isolated from the Antarctic marine waters and a penguin rookery, for 2 reasons: (i) as a measure of human impact and (ii) as a potential "snapshot" of the preantibiotic world. Samples were taken at 4 established sampling sites near Palmer Station, which is situated at the southern end of the Palmer Archipelago (64 degrees 10'S, 61 degrees 50'W). Sites were chosen to provide different potentials for human contamination. Forty 50 mL samples of seawater were collected and colony-forming units (CFU)/mL were determined at 6 and 20 degrees C. For this study, presumed psychrophiles (growth at 6 degrees C) were assumed to be native to Antarctic waters, whereas presumed mesophiles (growth at 20 degrees C but not at 6 degrees C) were taken to represent introduced organisms. The 20-6 degrees C CFU/mL ratio was used as a measure of the relative impact to the ecosystem of presumably introduced organisms. This ratio was highest at the site nearest to Palmer Station and decreased with distance from it, suggesting that human presence has impacted the natural microbial flora of the site. The frequency of resistance to 5 common antibiotics was determined in each group of isolates. Overall drug resistance was higher among the presumed mesophiles than the presumed psychrophiles and increased with proximity to Palmer Station, with the presumed mesophiles showing higher frequencies of single and multiple drug resistance than the psychrophile population. The frequency of multidrug resistance followed the same pattern. It appears that multidrug resistance is low among native Antarctic bacteria but is increased by human habitation.

  10. Loading of fecal indicator bacteria in North Carolina tidal creek headwaters: hydrographic patterns and terrestrial runoff relationships.

    PubMed

    Stumpf, Curtis H; Piehler, Michael F; Thompson, Suzanne; Noble, Rachel T

    2010-09-01

    In the New River Estuary (NRE) in eastern North Carolina (NC), fecal indicator bacteria (FIB) levels exceed water quality standards, leading to closure of estuarine waters for shellfishing and classification of parts of the estuary as "impaired" per the Clean Water Act section 303(d) list. As a means to investigate fecal contamination and loading of FIB to the NRE, a continuous automated sampler (ISCO) outfitted with flow modules and water quality probes was placed in four first-order tidal creek headwaters. Total storm discharge and bacterial load for Escherichia coli (EC) and Enterococcus spp. (ENT) were calculated using graphical volumetric flow calculations and interpolation of FIB measurements over each storm's duration for 10 storms. Mean total load of 10(9)-10(12) EC and ENT cells (MPN) occurred over the course of each storm. Total storm loading, averaged across all storms, was as much as 30 and 37 times greater than equivalent duration of baseflow loading for EC and ENT, respectively. Within the first 30% of creek storm volume for all storms and all creeks combined, a mean cumulative load of only 37% and 44% of the total EC and ENT cells, respectively, was discharged, indicating these creeks are not demonstrating a 'first flush' scenario for FIB. The median storm Event Mean Concentrations (EMCs) were 6.37 × 10(2) and 2.03 × 10(2) MPN/100 mL, for EC and ENT, respectively, compared with median baseflow concentrations of 1.48 × 10(2) and 4.84 × 10(1) for EC and ENT, respectively, and were significantly different between base and storm flow events. FIB was correlated with TSS (weak), flow rate (strong), and different stages (base, rising, peak, and falling) of the hydrograph (strong). Pollutographs indicate large intra-storm variability of FIB, and the need for more intensive sampling throughout a storm in order to attain accurate FIB contaminant estimates. Instream sediment concentrations ranged from 5 to 478 (MPN/g) and 13 to 776 (MPN/g) for EC and ENT

  11. Bacteria holding times for fecal coliform by mFC agar method and total coliform and Escherichia coli by Colilert®-18 Quanti-Tray® method

    USGS Publications Warehouse

    Aulenbach, Brent T.

    2010-01-01

    Bacteria holding-time experiments of up to 62 h were performed on five surface-water samples from four urban stream sites in the vicinity of Atlanta, GA, USA that had relatively high densities of coliform bacteria (Escherichia coli densities were all well above the US Environmental Protection Agency criterion of 126 colonies (100 ml) − 1 for recreational waters). Holding-time experiments were done for fecal coliform using the membrane filtration modified fecal coliform (mFC) agar method and for total coliform and E. coli using the Colilert®-18 Quanti-Tray® method. The precisions of these analytical methods were quantified. Precisions determined for fecal coliform indicated that the upper bound of the ideal range of counts could reasonably be extended upward and would improve precision. For the Colilert®-18 method, analytical precisions were similar to the theoretical precisions for this method. Fecal and total coliform densities did not change significantly with holding times up to about 27 h. Limited information indicated that fecal coliform densities might be stable for holding times of up to 62 h, whereas total coliform densities might not be stable for holding times greater than about 27 h. E. coli densities were stable for holding times of up to 18 h—a shorter period than indicated from a previous studies. These results should be applicable to non-regulatory monitoring sampling designs for similar urban surface-water sample types.

  12. Effect of Sunlight on the Divergence of Community Structure of Fecal Bacteria in Cowpats Collected from Three Different Farms

    EPA Science Inventory

    Fecal pollution of environmental waters is a major concern for the general public because exposure to fecal-associated pathogens can have severe impacts on human health. In the last few years, numerous metagenomic studies applied next generation sequencing to understand the shift...

  13. Effect of Sunlight on the Divergence of Community Structure of Fecal Bacteria in Cowpats Collected from Three Different Farms

    EPA Science Inventory

    Fecal pollution of environmental waters is a major concern for the general public because exposure to fecal-associated pathogens can have severe impacts on human health. In the last few years, numerous metagenomic studies applied next generation sequencing to understand the shift...

  14. Thermotolerant non-fecal source Klebsiella pneumoniae: validity of the fecal coliform test in recreational waters.

    PubMed Central

    Caplenas, N R; Kanarek, M S

    1984-01-01

    Wisconsin pulp and paper mill processing plants were evaluated for fecal coliform and total Klebsiella (i.e., thermotolerant and thermointolerant) bacterial concentrations. Using the standard fecal coliform test, up to 90 per cent of non-fecal source thermotolerant K. pneumoniae was falsely identified as fecal source bacteria. Since there is a lack of specificity in the currently used standard for fecal coliform evaluation, a more reliable health risk assessment for fecal coliform bacteria is recommended. PMID:6388365

  15. Effects of different sources of fructans on body weight, blood metabolites and fecal bacteria in normal and obese non-diabetic and diabetic rats.

    PubMed

    Rendón-Huerta, Juan A; Juárez-Flores, Bertha; Pinos-Rodríguez, Juan M; Aguirre-Rivera, J Rogelio; Delgado-Portales, Rosa E

    2012-03-01

    Fructans contribute significantly to dietary fiber with beneficial effects on gastrointestinal physiology in healthy individuals and offer a promising approach to treating some diseases. Two experiments (Experiment 1 = rats with normal weight; Experiment 2 = obese rats) were developed to compare the effects of three fructan sources (Cichorium intybus L. Asteraceae, Helianthus tuberosus L. Asteraceae and Agave angustifolia ssp. tequilana Haw, Agavaceae) on body weight change, blood metabolites and fecal bacteria in non-diabetic (ND) and diabetic (D) rats. In Experiment 1 total body weight gain and daily feed intake in D and ND rats decreased (P < 0.05) with supplements of fructan. Only in D rats, blood glucose concentrations, fecal Clostrodium spp. counts, and liver steatosis decreased, while blood HDL concentrations and fecal Lactobacillus spp. and Bifidobacterium spp. counts increased due to fructans. In Experiment 2, total body weight gain and feed intake in ND and D rats were also decreased by fructans. In ND rats, fructan decreased blood glucose concentrations. In D rats, fructans from A. angustifolia ssp. tequilana decreased blood cholesterol and LDL and liver steatosis. For both ND and D rats, fecal Lactobacillus spp. and Bifidobacterium spp. counts were higher (P < 0.05) with fructan supplements.

  16. Persistence of fecal indicator bacteria in sediment of an oligotrophic river: comparing large and lab-scale flume systems.

    PubMed

    Walters, Evelyn; Kätzl, Korbinian; Schwarzwälder, Kordula; Rutschmann, Peter; Müller, Elisabeth; Horn, Harald

    2014-09-15

    In this study, both a lab and a large-scale flume system were used to investigate the survival of fecal indicator bacteria (FIB) in bed sediments of an alpine oligotrophic river. To determine the influence of substratum on persistence, survival within 3-cm-deep substratum cages versus on thin, biofilm-covered ceramic tiles was tested. Moreover, the impact of bed shear stress on survival in bed sediments was explored. It was seen that in the lab-scale flume having a very low bed shear stress (0.3 N m(-2)), E. coli and enterococci survival in 3-cm-deep substratum cages was nearly the same as in a thin biofilm (200 μm). However, in the large-scale flume system characterized by a bed shear stress of 9 N m(-2), the added protection of the deeper substratum cages promoted considerably longer survival of E. coli and enterococci than the thin biofilm. Additionally, the FIB removal mechanisms in the two flume systems varied. At the lab-scale, enterococci was seen to persist twice as long as E. coli, while in the large-scale flume the two FIB were removed at the same rate. A comparison of qPCR analyses performed in both flumes suggests that bed sediment erosion and the influence of grazers/predators were responsible for FIB removal from the sediments in the large-scale flume, whereas in the lab flume FIB inactivation caused removal. These results indicate that hydraulic parameters such as bed shear stress as well as the presence of macroinvertebrates in a system are both important factors to consider when designing flumes as they can significantly impact FIB persistence in sediments of fast-flowing, alpine streams.

  17. Evaluation of methods to sample fecal indicator bacteria in foreshore sand and pore water at freshwater beaches.

    PubMed

    Vogel, Laura J; Edge, Thomas A; O'Carroll, Denis M; Solo-Gabriele, Helena M; Kushnir, Caitlin S E; Robinson, Clare E

    2017-09-15

    Fecal indicator bacteria (FIB) are known to accumulate in foreshore beach sand and pore water (referred to as foreshore reservoir) where they act as a non-point source for contaminating adjacent surface waters. While guidelines exist for sampling surface waters at recreational beaches, there is no widely-accepted method to collect sand/sediment or pore water samples for FIB enumeration. The effect of different sampling strategies in quantifying the abundance of FIB in the foreshore reservoir is unclear. Sampling was conducted at six freshwater beaches with different sand types to evaluate sampling methods for characterizing the abundance of E. coli in the foreshore reservoir as well as the partitioning of E. coli between different components in the foreshore reservoir (pore water, saturated sand, unsaturated sand). Methods were evaluated for collection of pore water (drive point, shovel, and careful excavation), unsaturated sand (top 1 cm, top 5 cm), and saturated sand (sediment core, shovel, and careful excavation). Ankle-depth surface water samples were also collected for comparison. Pore water sampled with a shovel resulted in the highest observed E. coli concentrations (only statistically significant at fine sand beaches) and lowest variability compared to other sampling methods. Collection of the top 1 cm of unsaturated sand resulted in higher and more variable concentrations than the top 5 cm of sand. There were no statistical differences in E. coli concentrations when using different methods to sample the saturated sand. Overall, the unsaturated sand had the highest amount of E. coli when compared to saturated sand and pore water (considered on a bulk volumetric basis). The findings presented will help determine the appropriate sampling strategy for characterizing FIB abundance in the foreshore reservoir as a means of predicting its potential impact on nearshore surface water quality and public health risk. Copyright © 2017 Elsevier Ltd. All rights

  18. Fecal Indicator Bacteria Entrainment from Streambed to Water Column: Transport by Unsteady Flow over a Sand Bed.

    PubMed

    Surbeck, Cristiane Q; Douglas Shields, F; Cooper, Alexandra M

    2016-05-01

    Storms cause a substantial increase in the fecal indicator bacteria (FIB) concentrations in stream water as a result of FIB-laden runoff and the release of FIB from stream sediments. Previous work has emphasized the association between FIB and bed sediments finer than sand. The objectives of this work were to elucidate the effect of various velocities on the entrainment of bed-dwelling coliforms in sand-bed streams and to refine methodologies for quantifying sandy streambeds as sources of FIB. Pump-induced hydrographs were created using a stainless steel nonrecirculating flume. Experiments consisted of simulating four storm hydrographs and collecting water samples upstream and downstream of a sand bed at selected intervals. Bed sediment samples were collected before and after each event. The highest concentrations of total coliform and suspended sediments generally occurred in the downstream samples during the rising limb of the hydrographs as a result of entrainment of coliforms and sand from the bed to the water column. There was a first flush effect in the system, as the upper layer of sand was influenced by a rapidly increasing velocity at ∼0.2 m s. Coliforms downstream of the sand bed increased rapidly as velocity exceeded this threshold but then declined even as velocity and discharge continued to increase. This likely reflects the depletion of coliforms as the more densely populated sediment layer was flushed out. There is evidence that streams with sand beds harbor enough FIB that development of total maximum daily loads (TMDLs) should include consideration of them as a source.

  19. Bacteroides fragilis type VI secretion systems use novel effector and immunity proteins to antagonize human gut Bacteroidales species.

    PubMed

    Chatzidaki-Livanis, Maria; Geva-Zatorsky, Naama; Comstock, Laurie E

    2016-03-29

    Type VI secretion systems (T6SSs) are multiprotein complexes best studied in Gram-negative pathogens where they have been shown to inhibit or kill prokaryotic or eukaryotic cells and are often important for virulence. We recently showed that T6SS loci are also widespread in symbiotic human gut bacteria of the order Bacteroidales, and that these T6SS loci segregate into three distinct genetic architectures (GA). GA1 and GA2 loci are present on conserved integrative conjugative elements (ICE) and are transferred and shared among diverse human gut Bacteroidales species. GA3 loci are not contained on conserved ICE and are confined to Bacteroides fragilis Unlike GA1 and GA2 T6SS loci, most GA3 loci do not encode identifiable effector and immunity proteins. Here, we studied GA3 T6SSs and show that they antagonize most human gut Bacteroidales strains analyzed, except for B. fragilis strains with the same T6SS locus. A combination of mutation analyses,trans-protection analyses, and in vitro competition assays, allowed us to identify novel effector and immunity proteins of GA3 loci. These proteins are not orthologous to known proteins, do not contain identified motifs, and most have numerous predicted transmembrane domains. Because the genes encoding effector and immunity proteins are contained in two variable regions of GA3 loci, GA3 T6SSs of the species B. fragilis are likely the source of numerous novel effector and immunity proteins. Importantly, we show that the GA3 T6SS of strain 638R is functional in the mammalian gut and provides a competitive advantage to this organism.

  20. Bacteroides fragilis type VI secretion systems use novel effector and immunity proteins to antagonize human gut Bacteroidales species

    PubMed Central

    Chatzidaki-Livanis, Maria; Geva-Zatorsky, Naama; Comstock, Laurie E.

    2016-01-01

    Type VI secretion systems (T6SSs) are multiprotein complexes best studied in Gram-negative pathogens where they have been shown to inhibit or kill prokaryotic or eukaryotic cells and are often important for virulence. We recently showed that T6SS loci are also widespread in symbiotic human gut bacteria of the order Bacteroidales, and that these T6SS loci segregate into three distinct genetic architectures (GA). GA1 and GA2 loci are present on conserved integrative conjugative elements (ICE) and are transferred and shared among diverse human gut Bacteroidales species. GA3 loci are not contained on conserved ICE and are confined to Bacteroides fragilis. Unlike GA1 and GA2 T6SS loci, most GA3 loci do not encode identifiable effector and immunity proteins. Here, we studied GA3 T6SSs and show that they antagonize most human gut Bacteroidales strains analyzed, except for B. fragilis strains with the same T6SS locus. A combination of mutation analyses, trans-protection analyses, and in vitro competition assays, allowed us to identify novel effector and immunity proteins of GA3 loci. These proteins are not orthologous to known proteins, do not contain identified motifs, and most have numerous predicted transmembrane domains. Because the genes encoding effector and immunity proteins are contained in two variable regions of GA3 loci, GA3 T6SSs of the species B. fragilis are likely the source of numerous novel effector and immunity proteins. Importantly, we show that the GA3 T6SS of strain 638R is functional in the mammalian gut and provides a competitive advantage to this organism. PMID:26951680

  1. Fecal-indicator bacteria and Escherichia coli pathogen data collected near a novel sub-irrigation water-treatment system in Lenawee County, Michigan, June-November 2007

    USGS Publications Warehouse

    Duris, Joseph W.; Beeler, Stephanie

    2008-01-01

    The U.S. Geological Survey, in cooperation with the Lenawee County Conservation District in Lenawee County, Mich., conducted a sampling effort over a single growing season (June to November 2007) to evaluate the microbiological water quality around a novel livestock reservoir wetland sub-irrigation system. Samples were collected and analyzed for fecal coliform bacteria, Escherichia coli (E. coli) bacteria, and six genes from pathogenic strains of E. coli.A total of 73 water-quality samples were collected on nine occasions from June to November 2007. These samples were collected within the surface water, shallow ground water, and the manure-treatment system near Bakerlads Farm near Clayton in Lenawee County, Mich. Fecal coliform bacteria concentrations ranged from 10 to 1.26 million colony forming units per 100 milliliters (CFU/100 mL). E. coli bacteria concentrations ranged from 8 to 540,000 CFU/100 mL. Data from the E. coli pathogen analysis showed that 73 percent of samples contained the eaeA gene, 1 percent of samples contained the stx2 gene, 37 percent of samples contained the stx1 gene, 21 percent of samples contained the rfbO157 gene, and 64 percent of samples contained the LTIIa gene.

  2. Fecal-coliform bacteria concentrations in streams of the Chattahoochee River National Recreation Area, Metropolitan Atlanta, Georgia, May-October 1994 and 1995

    USGS Publications Warehouse

    Gregory, M. Brian; Frick, Elizabeth A.

    2000-01-01

    Introduction: The Metropolitan Atlanta area has been undergoing a period of rapid growth and development. The population in the 10-county metropolitan area almost doubled from about 1.5 million people in 1970 to 2.9 million people in 1995 (Atlanta Regional Commission, written commun., 2000). Residential, commercial, and other urban land uses more than tripled during the same period (Frick and others, 1998). The Chattahoochee River is the most utilized water resource in Georgia. The rapid growth of Metropolitan Atlanta and its location downstream of the headwaters of the drainage basin make the Chattahoochee River a vital resource for drinking-water supplies, recreational opportunities, and wastewater assimilation. In 1978, the U.S. Congress declared the natural, scenic, recreation, and other values of 48 miles of the Chattahoochee River from Buford Dam to Peachtree Creek to be of special national significance. To preserve this reach of the Chattahoochee River, the U.S. Congress created the Chattahoochee River National Recreational Area (CRNRA), which includes the Chattahoochee River downstream from Buford Dam to the mouth of Peachtree Creek and a series of park areas adjacent to the river in northern Metropolitan Atlanta Even with this protection, waters of the Chattahoochee River and many of its tributaries in Metropolitan Atlanta did not meet water-quality standards set for designated uses during 1994 and 1995 (fig. 1 and table 1). Much of the degradation of water quality has been associated with areas undergoing rapid urban growth and sprawling suburban development. The resulting conversion of mostly forested land to urban land has multiple adverse effects on water quality. Degradation of water quality may be caused by a number of factors including an increase in nutrient concentrations, sediment and sedimentbound contaminant concentrations (e.g., metals and pesticides) (Frick and others, 1998), and fecal-coliform bacteria concentrations (Center for Watershed

  3. Identifying fecal sources in a selected catchment reach using multiple source-tracking tools

    USGS Publications Warehouse

    Vogel, J.R.; Stoeckel, D.M.; Lamendella, R.; Zelt, R.B.; Santo, Domingo J.W.; Walker, S.R.; Oerther, D.B.

    2007-01-01

    Given known limitations of current microbial source-tracking (MST) tools, emphasis on small, simple study areas may enhance interpretations of fecal contamination sources in streams. In this study, three MST tools - Escherichia coli repetitive element polymerase chain reaction (rep-PCR), coliphage typing, and Bacteroidales 16S rDNA host-associated markers - were evaluated in a selected reach of Plum Creek in sooth-central Nebraska. Water-quality samples were collected from six sites. One reach was selected for MST evaluation based on observed patterns of E. coli contamination. Despite high E. coli concentrations, coliphages were detected only once among water samples, precluding their use as a MST tool in this setting. Rep-PCR classification of E. coli isolates from both water and sediment samples supported the hypothesis that cattle and wildlife were dominant sources of fecal contamination, with minor contributions by horses and humans. Conversely, neither ruminant nor human sources were detected by Bacteroidales markers in most water samples. In bed sediment, ruminant- and human-associated Bacteroidales markers were detected throughout the interval from 0 to 0.3 m, with detections independent of E. coli concentrations in the sediment. Although results by E. coli-based and Bacteroidales-based MST methods led to similar interpretations, detection of Bacteroidales markers in sediment more commonly than in water indicates that different tools to track fecal contamination (in this case, tools based on Bacteroidales DNA and E. coli isolates) may have varying relevance to the more specific goal of tracking the sources of E. coli in watersheds. This is the first report of simultaneous, toolbox approach application of a library-based and marker-based MST analyses to lowing surface water. ?? ASA, CSSA, SSSA.

  4. Comparison of fecal coliform bacteria before and after wastewater treatment plant in the Izmir Bay (Eastern Aegean Sea).

    PubMed

    Kacar, Asli; Gungor, Fatma

    2010-03-01

    The distribution of fecal coliforms was investigated and determined in Izmir Bay from 1996 to 2005. Izmir Bay severely was polluted from industrial and domestic discharges during decades. In early 2000, a wastewater treatment plant began to treat domestic and industrial wastes. This plant treats the wastes about 80% capacity after 2001. The sampling periods cover before and after treatment plant. Assessment method for determining the number of fecal coliform has evolved membrane filtrations. Maximum surface fecal coliform concentration was 4.9 x 10(5) cfu 100 ml(-1) in 1996-2000 period. Following the opening treatment system, fecal coliform density decreased 2.1 x 10(4) cfu 100 ml(-1) during 2001-2005. A continuous improvement can be sustained in the water quality if direct inflow of untreated wastewater is prevented.

  5. Interacting effects of sunlight, agriculturally derived dissolved organic matter and reactive oxygen species on fecal indicator bacteria growth dynamics

    EPA Science Inventory

    Bacterial survival in agriculturally impacted surface waters is dependent on resource availability and also on potential resource transformations, mediated by biotic and abiotic processes. In this study, we focused on the effect of sunlight irradiated cattle fecal extract (CFE) a...

  6. Preliminary survey of antibiotic-resistant fecal indicator bacteria and pathogenic Escherichia coli from river-water samples collected in Oakland County, Michigan, 2003

    USGS Publications Warehouse

    Fogarty, Lisa R.; Duris, Joseph W.; Aichele, Stephen S.

    2005-01-01

    A preliminary study was done in Oakland County, Michigan, to determine the concentration of fecal indicator bacteria (fecal coliform bacteria and enterococci), antibiotic resistance patterns of these two groups, and the presence of potentially pathogenic Escherichia coli (E. coli). For selected sites, specific members of these groups [E. coli, Enterococcus faecium (E. faecium) and Enterococcus faecalis (E. faecalis)] were isolated and tested for levels of resistance to specific antibiotics used to treat human infections by pathogens in these groups and for their potential to transfer these resistances. In addition, water samples from all sites were tested for indicators of potentially pathogenic E. coli by three assays: a growth-based assay for sorbitol-negative E. coli, an immunological assay for E. coli O157, and a molecular assay for three virulence and two serotype genes. Samples were also collected from two non-urbanized sites outside of Oakland County. Results from the urbanized Oakland County area were compared to those from these two non-urbanized sites. Fecal indicator bacteria concentrations exceeded State of Michigan recreational water-quality standards and (or) recommended U.S. Environmental Protection Agency (USEPA) standards in samples from all but two Oakland County sites. Multiple-antibiotic-resistant fecal coliform bacteria were found at all sites, including two reference sites from outside the county. Two sites (Stony Creek and Paint Creek) yielded fecal coliform isolates resistant to all tested antibiotics. Patterns indicative of extended-spectrum-β-lactamase (ESBL)- producing fecal coliform bacteria were found at eight sites in Oakland County and E. coli resistant to clinically significant antibiotics were recovered from the River Rouge, Clinton River, and Paint Creek. Vancomycin-resistant presumptive enterococci were found at six sites in Oakland County and were not found at the reference sites. Evidence of acquired antibiotic resistances was

  7. Comparisons of statistical models to predict fecal indicator bacteria concentrations enumerated by qPCR- and culture-based methods.

    PubMed

    Gonzalez, Raul A; Noble, Rachel T

    2014-01-01

    Recently, the United States Environmental Protection Agency (USEPA) revised their recreational water quality criteria, in which adjustments were made by approving enterococci (ENT) quantitative PCR (qPCR) as an alternative, rapid method and advocating the use of predictive models for water quality management. The implementation of qPCR-based methods and prediction models are meant to decrease the time between sample collection and public advisories and notifications. To date, few studies have compared qPCR-based models to culture-based prediction models and none of these studies have been conducted in coastal estuarine systems. In this study, we created prediction models using qPCR-based fecal indicator bacteria (FIB) data in dual-use recreational and shellfish harvesting waters and compared them to published ENT and Escherichia coli (EC) culture-based prediction models in eastern North Carolina estuaries. Furthermore, an empirical statistical model was created to predict qPCR inhibition levels so that proper remediation techniques can be applied when it is a problem. Predictor variable selection in both qPCR- and culture-based ENT models was very similar; both models included 14-day rain total, dissolved oxygen, and salinity/conductivity, with 89 and 90% of qPCR and culture data described, respectively. Using ENT management action thresholds, qPCR- and culture-based models showed high accuracy in management decisions. The qPCR model had 92 and 96% accuracy using the 110 and 1000 cell equivalents (CE)/100 ml thresholds, respectively, and the culture model had 90% accuracy in management decisions with the 110 MPN/100 ml threshold. EC models for qPCR- and culture-based concentrations used similar independent variables (14-day humidity, salinity/conductivity, a rain/storm variable, and a measure of air temperature), with each model explaining 26 and 55% of the data variation, respectively. When using different thresholds that were logs apart for management decisions

  8. Evaluation of potential sources and transport mechanisms of fecal indicator bacteria to beach water, Murphy Park Beach, Door County, Wisconsin

    USGS Publications Warehouse

    Juckem, Paul F.; Corsi, Steven R.; McDermott, Colleen; Kleinheinz, Gregory; Fogarty, Lisa R.; Haack, Sheridan K.; Johnson, Heather E.

    2013-01-01

    Fecal Indicator Bacteria (FIB) concentrations in beach water have been used for many years as a criterion for closing beaches due to potential health concerns. Yet, current understanding of sources and transport mechanisms that drive FIB occurrence remains insufficient for accurate prediction of closures at many beaches. Murphy Park Beach, a relatively pristine beach on Green Bay in Door County, Wis., was selected for a study to evaluate FIB sources and transport mechanisms. Although the relatively pristine nature of the beach yielded no detection of pathogenic bacterial genes and relatively low FIB concentrations during the study period compared with other Great Lakes Beaches, its selection limited the number of confounding FIB sources and associated transport mechanisms. The primary sources of FIB appear to be internal to the beach rather than external sources such as rivers, storm sewer outfalls, and industrial discharges. Three potential FIB sources were identified: sand, swash-zone groundwater, and Cladophora mats. Modest correlations between FIB concentrations in these potential source reservoirs and FIB concentrations at the beach from the same day illustrate the importance of understanding transport mechanisms between FIB sources and the water column. One likely mechanism for transport and dispersion of FIB from sand and Cladophora sources appears to be agitation of Cladophora mats and erosion of beach sand due to storm activity, as inferred from storm indicators including turbidity, wave height, current speed, wind speed, sky visibility, 24-hour precipitation, and suspended particulate concentration. FIB concentrations in beach water had a statistically significant relation (p-value ‹0.05) with the magnitude of these storm indicators. In addition, transport of FIB in swash-zone groundwater into beach water appears to be driven by groundwater recharge associated with multiday precipitation and corresponding increased swash-zone groundwater discharge at

  9. Hormones, sterols, and fecal indicator bacteria in groundwater, soil, and subsurface drainage following a high single application of municipal biosolids to a field.

    PubMed

    Gottschall, N; Topp, E; Edwards, M; Payne, M; Kleywegt, S; Russell, P; Lapen, D R

    2013-04-01

    A land application of dewatered municipal biosolids (DMB) was conducted on an agricultural field in fall 2008 at a rate of 22Mg dry weight (dw) ha(-1). Pre- and post- application, hormone, sterol and fecal indicator bacteria concentrations were measured in tile drainage water, groundwater (2, 4, 6m depth), surface soil cores, and DMB aggregates incorporated in the soil (∼0.2m depth) for a period of roughly 1year post-application. Hormones and sterols were detected up to 1year post-application in soil and in DMB aggregates. Hormone (androsterone, desogestrel, estrone) contamination was detected briefly in tile water samples (22d and ∼2months post-app), at lowngL(-1) concentrations (2-34ngL(-1)). Hormones were not detected in groundwater. Sterols were detected in tile water throughout the study period post-application, and multiple fecal sterol ratios suggested biosolids as the source. Coprostanol concentrations in tile water peaked at >1000ngL(-1) (22d post-app) and were still >100ngL(-1) at 6months post-application. Fecal indicator bacteria were detected throughout the study period in tile water, groundwater (⩽2m depth), soil and DMB aggregate samples. These bacteria were strongly linearly related to coprostanol in tile water (R(2)>0.92, p<0.05). The limited transport of hormones and sterols to tile drainage networks may be attributed to a combination of the hydrophobicity of these compounds and limited macroporosity of the field soil. This transitory contamination from hormones and sterols is unlikely to result in any significant pulse exposure risk in subsurface drainage and groundwater. Crown Copyright © 2012. Published by Elsevier Ltd. All rights reserved.

  10. Identification of lactic acid bacteria isolated from the fecal samples of healthy humans and patients with dyspepsia, and determination of their ph, bile, and antibiotic tolerance properties.

    PubMed

    Kiliç, Gülden Başyiğit; Karahan, Aynur Gül

    2010-01-01

    The research of novel probiotic strains is important in order to satisfy the increasing request of the market and to obtain functional products in which the probiotic cultures are more active and have better probiotic characteristics than those already on the market. In this study, the probiotic potential of Lactobacillus and Enterococcus strains isolated from human feces was investigated. One hundred seven strains of lactic acid bacteria were isolated from the fecal samples of 19 adult volunteers with stomach problems and the fecal samples of 12 healthy adult volunteers. The strains were identified by 16S rRNA analysis and examined for resistance to gastric acidity (pH 3.5) and bile salts (0.3% bile salt). The antibiotic susceptibility of the strains was also determined. The counts of lactic acid bacteria in the fecal samples ranged from 1.7 x 10(7) to 3.1 x 10(10) CFU/g. The species present in those samples were found to be Lactobacillusplantarum, L. fermentum, Enterococcus faecium and E. durans. Results indicated that L. plantarum (AA1-2, AA17-73, AC18-88, AK4-11, and AK7-28), L. fermentum (AB5-18, BB16-75, and AK4-180), E. faecium (AB20-98 and BK11-50) and E. durans (AK4-14 and BK9-40) showed good pH and bile tolerance. These strains are potentially good candidates for use as health-promoting bacteria.

  11. Antibiotic resistance patterns in fecal bacteria isolated from Christmas shearwater (Puffinus nativitatis) and masked booby (Sula dactylatra) at remote Easter Island.

    PubMed

    Ardiles-Villegas, Karen; González-Acuña, Daniel; Waldenström, Jonas; Olsen, Björn; Hernández, Jorge

    2011-09-01

    Antibiotic use and its implications have been discussed extensively in the past decades. This situation has global consequences when antibiotic resistance becomes widespread in the intestinal bacterial flora of stationary and migratory birds. This study investigated the incidence of fecal bacteria and general antibiotic resistance, with special focus on extended spectrum beta-lactamase (ESBL) isolates, in two species of seabirds at remote Easter Island. We identified 11 species of bacteria from masked booby (Sula dactylatra) and Christmas shearwater (Puffinus nativitatis); five species of gram-negative bacilli, four species of Streptococcus (Enterococcus), and 2 species of Staphylococcus. In addition, 6 types of bacteria were determined barely to the genus level. General antibiotic susceptibility was measured in the 30 isolated Enterobacteriaceae to 11 antibiotics used in human and veterinary medicine. The 10 isolates that showed a phenotypic ESBL profile were verified by clavulanic acid inhibition in double mixture discs with cefpodoxime, and two ESBL strains were found, one strain in masked booby and one strain in Christmas shearwater. The two bacteria harboring the ESBL type were identified as Serratia odorifera biotype 1, which has zoonotic importance. Despite minimal human presence in the masked booby and Christmas shearwater habitats, and the extreme geographic isolation of Easter Island, we found several multiresistant bacteria and even two isolates with ESBL phenotypes. The finding of ESBLs has animal and public health significance and is of potential concern, especially because the investigation was limited in size and indicated that antibiotic-resistant bacteria now are distributed globally.

  12. Factors affecting the presence of human-associated and fecal indicator real-time quantitative PCR genetic markers in urban-impacted recreational beaches.

    PubMed

    Molina, Marirosa; Hunter, Shayla; Cyterski, Mike; Peed, Lindsay A; Kelty, Catherine A; Sivaganesan, Mano; Mooney, Thomas; Prieto, Lourdes; Shanks, Orin C

    2014-11-01

    Urban runoff can carry a variety of pollutants into recreational beaches, often including bacterial pathogens and indicators of fecal contamination. To develop complete recreational criteria and risk assessments, it is necessary to understand conditions under which human contamination could be present at beaches solely impacted by urban runoff. Accurately estimating risk requires understanding sources, concentrations, and transport mechanisms of microbial contaminants in these environments. By applying microbial source tracking methods and empirical modeling, we assessed the presence and level of human contamination at urban runoff impacted recreational beaches. We also identified environmental parameters and pollution sources that can influence the concentration and transport of culturable and molecular fecal indicator bacteria (FIB) in systems impacted solely by urban runoff. Water samples and physico-chemical parameters were collected from shoreline locations from three South Carolina (SC) beaches (five locations per beach) and two Florida (FL) beaches (three locations per beach). Each SC beach was directly impacted by swashes or tidal creeks receiving stormwater runoff from the urbanized area and therefore were designated as swash drain associated (SDA) beaches, while FL beaches were designated as non-swash drain associated (NSDA). Sampling in swash drains (SD; three sites per SD) directly impacting each SC beach was also conducted. Results indicate that although culturable (enterococci) and real-time quantitative polymerase chain reaction (qPCR) (EC23S857, Entero1, and GenBac3) FIB concentrations were, on average, higher at SD locations, SDA beaches did not have consistently higher molecular FIB signals compared to NSDA beaches. Both human-associated markers (HF183 and HumM2) were concomitantly found only at SDA beaches. Bacteroidales species-specific qPCR markers (BsteriF1 and BuniF2) identified differences in the Bacteroidales community, depending on beach

  13. Evaluation of Colilert-18 for detection and enumeration of fecal coliform bacteria in wastewater using the U.S. Environmental Protection Agency Alternative Test procedure Protocol.

    PubMed

    Warden, Paul S; DeSarno, Monique S; Volk, Sarah E; Eldred, Bradley J

    2011-01-01

    This study compared recovery of fecal coliform bacteria from sewage by Colilert-18 and Standard Methods 9222D (membrane-Fecal Coliform medium) in accordance with the U.S. Environmental Protection Agency (EPA) Alternative Test Protocol (ATP). Samples were collected from 10 different wastewater treatment plants in the northeastern United States and tested in a single laboratory. Twenty replicates of each sample were analyzed by each method, and 200 positive and 200 negative responses were confirmed for each method. Recovery of fecal coliforms by Colilert-18 was significantly higher than (8 of 10 sites) or statistically equivalent to (1 of 10 sites) recovery by the reference method (Standard Methods 9222D) for samples from all but one site. Both methods had low false-positive rates (< 2%); however, the false-negative rate observed with Standard Methods 9222D (21.5%) was substantially higher than that observed with Colilert-18 (7%). The accuracy rates of the two methods were calculated as 96.5 and 88.9% for Colilert-18 and Standard Methods 9222D, respectively. The results of this study demonstrate that Colilert-18 meets the acceptance criteria for alternative methods specified in the EPA ATP.

  14. Molecular detection by analysis of the 16S rRNA gene of fecal coliform bacteria from the two Korean Apodemus species (Apodemus agrarius and A. peninsulae).

    PubMed

    Rahman, M M; Yoon, K B; Lim, S J; Jeon, M G; Kim, H J; Kim, H Y; Cho, J Y; Chae, H M; Park, Y C

    2017-05-18

    Wild mouse feces can disseminate zoonotic microorganisms throughout a farm, which is a great threat to human health and can lead to economic loss through contaminated agricultural produce. To assess the microbial communities, especially fecal coliform bacteria, we used two methods. First, we isolated bacterial colonies onto the common media LB (lactose broth) agar, TSA (tryptic soy agar), and MRS (de Man, Rogosa, and Sharpe) agar, and then randomly select colonies from each plate and stocked them to the mother plate for genomic DNA isolation. Second, we analyzed bacterial colonies using the 16S rRNA gene molecular diagnostic method. Based on bacterial cultures and bacterial 16S rRNA gene markers, we detected four different bacterial species (Bacillus amyloliquefaciens, Escherichia coli, Staphylococcus xylosus, and Serratia liquefaciens) from fecal coliforms of the striped field mouse Apodemus agrarius and A. peninsulae in agricultural areas in South Korea. These results could help us to better understand the pathogen reservoirs of mice and initiate some preventive measures to mitigate the microbial risks associated with mouse fecal matter in agricultural production areas.

  15. Run-off studies demonstrate parallel transport behaviour for a marker of poultry fecal contamination and Staphylococcus aureus.

    PubMed

    Weidhaas, J; Garner, E; Basden, T; Harwood, V J

    2014-08-01

    To determine whether poultry litter marker gene LA35 is correlated with pathogens and fecal indicator bacteria (FIB) in run-off from poultry litter-amended plots. A rainfall simulator with various vegetative filter strip lengths was employed to evaluate the correlation of a microbial source tracking (MST) marker for poultry feces/litter (the 16S rRNA gene of Brevibacterium sp. LA35 [LA35] measured by quantitative PCR) with pathogens and FIB in run-off. LA35 was correlated with Staphylococcus aureus, Escherichia coli, Enterococcus spp. and Bacteroidales levels. Salmonella was present at low concentration in litter, but became undetectable by qPCR in run-off. Escherichia coli, LA35 and Staph. aureus exhibited mass-based first flush behaviour in the run-off. Correlation of LA35 with FIB and pathogens in run-off from poultry litter-amended fields suggest comparable transport mechanisms and that LA35 is a useful tracer for harmful bacteria in the environment released from poultry litter. To protect human health, an effective marker for poultry fecal contamination should exhibit similar fate and transport characteristics compared to pathogens. This study is among the first to demonstrate such a relationship in run-off for a MST marker. © 2014 The Society for Applied Microbiology.

  16. Theoretical and Numerical Modeling of Transport of Land Use-Specific Fecal Source Identifiers

    NASA Astrophysics Data System (ADS)

    Bombardelli, F. A.; Sirikanchana, K. J.; Bae, S.; Wuertz, S.

    2008-12-01

    Microbial contamination in coastal and estuarine waters is of particular concern to public health officials. In this work, we advocate that well-formulated and developed mathematical and numerical transport models can be combined with modern molecular techniques in order to predict continuous concentrations of microbial indicators under diverse scenarios of interest, and that they can help in source identification of fecal pollution. As a proof of concept, we present initially the theory, numerical implementation and validation of one- and two-dimensional numerical models aimed at computing the distribution of fecal source identifiers in water bodies (based on Bacteroidales marker DNA sequences) coming from different land uses such as wildlife, livestock, humans, dogs or cats. These models have been developed to allow for source identification of fecal contamination in large bodies of water. We test the model predictions using diverse velocity fields and boundary conditions. Then, we present some preliminary results of an application of a three-dimensional water quality model to address the source of fecal contamination in the San Pablo Bay (SPB), United States, which constitutes an important sub-embayment of the San Francisco Bay. The transport equations for Bacteroidales include the processes of advection, diffusion, and decay of Bacteroidales. We discuss the validation of the developed models through comparisons of numerical results with field campaigns developed in the SPB. We determine the extent and importance of the contamination in the bay for two decay rates obtained from field observations, corresponding to total host-specific Bacteroidales DNA and host-specific viable Bacteroidales cells, respectively. Finally, we infer transport conditions in the SPB based on the numerical results, characterizing the fate of outflows coming from the Napa, Petaluma and Sonoma rivers.

  17. Decay of bacterial pathogens, fecal indicators, and real-time quantitative PCR genetic markers in manure-amended soils.

    PubMed

    Rogers, Shane W; Donnelly, Matthew; Peed, Lindsay; Kelty, Catherine A; Mondal, Sumona; Zhong, Zirong; Shanks, Orin C

    2011-07-01

    This study examined persistence and decay of bacterial pathogens, fecal indicator bacteria (FIB), and emerging real-time quantitative PCR (qPCR) genetic markers for rapid detection of fecal pollution in manure-amended agricultural soils. Known concentrations of transformed green fluorescent protein-expressing Escherichia coli O157:H7/pZs and red fluorescent protein-expressing Salmonella enterica serovar Typhimurium/pDs were added to laboratory-scale manure-amended soil microcosms with moisture contents of 60% or 80% field capacity and incubated at temperatures of -20°C, 10°C, or 25°C for 120 days. A two-stage first-order decay model was used to determine stage 1 and stage 2 first-order decay rate coefficients and transition times for each organism and qPCR genetic marker in each treatment. Genetic markers for FIB (Enterococcus spp., E. coli, and Bacteroidales) exhibited decay rate coefficients similar to that of E. coli O157:H7/pZs but not of S. enterica serovar Typhimurium/pDs and persisted at detectable levels longer than both pathogens. Concentrations of these two bacterial pathogens, their counterpart qPCR genetic markers (stx1 and ttrRSBCA, respectively), and FIB genetic markers were also correlated (r = 0.528 to 0.745). This suggests that these qPCR genetic markers may be reliable conservative surrogates for monitoring fecal pollution from manure-amended land. Host-associated qPCR genetic markers for microbial source tracking decayed rapidly to nondetectable concentrations, long before FIB, Salmonella enterica serovar Typhimurium/pDs, and E. coli O157:H7/pZs. Although good indicators of point source or recent nonpoint source fecal contamination events, these host-associated qPCR genetic markers may not be reliable indicators of nonpoint source fecal contamination events that occur weeks following manure application on land.

  18. MONITORING FECAL INDICATOR BACTERIA WITH ALTERNATIVE REAL-TIME PCR INSTRUMENTS TO ASSESS HEALTH RISKS ASSOCIATED WITH RECREATIONAL WATER USE

    EPA Science Inventory

    U.S. EPA guidance on the safety of surface waters for recreational use is currently based on epidemiological studies conducted in the 1980?s that demonstrated a strong positive correlation between bathing-associated illness rates and concentrations of culturable fecal indicator b...

  19. USE OF COMPETITIVE GENOMIC HYBRIDIZATION TO ENRICH FOR GENOME-SPECIFIC DIFFERENCES BETWEEN TWO CLOSELY RELATED HUMAN FECAL INDICATOR BACTERIA

    EPA Science Inventory

    Enterococci are frequently used as indicators of fecal pollution in surface waters. To accelerate the identification of Enterococcus faecalis-specific DNA sequences, we employed a comparative genomic strategy utilizing a positive selection process to compare E. faec...

  20. Comparison of the New Millipore HC with Conventional Membrane Filters for the Enumeration of Fecal Coliform Bacteria

    PubMed Central

    Green, Barbara L.; Clausen, Elizabeth; Litsky, Warren

    1975-01-01

    Fecal coliform recoveries were determined for six types of membrane filters using 65 nonchlorinated water samples. Results showed that the membranes could be ranked in order of decreasing recovery as follows: Millipore HC > Gelman > Johns-Manville ∼ Sartorius > Millipore HA > Schleicher & Schuell. PMID:1103735

  1. USE OF COMPETITIVE GENOMIC HYBRIDIZATION TO ENRICH FOR GENOME-SPECIFIC DIFFERENCES BETWEEN TWO CLOSELY RELATED HUMAN FECAL INDICATOR BACTERIA

    EPA Science Inventory

    Enterococci are frequently used as indicators of fecal pollution in surface waters. To accelerate the identification of Enterococcus faecalis-specific DNA sequences, we employed a comparative genomic strategy utilizing a positive selection process to compare E. faec...

  2. MONITORING FECAL INDICATOR BACTERIA WITH ALTERNATIVE REAL-TIME PCR INSTRUMENTS TO ASSESS HEALTH RISKS ASSOCIATED WITH RECREATIONAL WATER USE

    EPA Science Inventory

    U.S. EPA guidance on the safety of surface waters for recreational use is currently based on epidemiological studies conducted in the 1980?s that demonstrated a strong positive correlation between bathing-associated illness rates and concentrations of culturable fecal indicator b...

  3. Estimated fecal coliform bacteria concentrations using near real-time continuous water-quality and streamflow data from five stream sites in Chester County, Pennsylvania, 2007–16

    USGS Publications Warehouse

    Senior, Lisa A.

    2017-09-15

    Several streams used for recreational activities, such as fishing, swimming, and boating, in Chester County, Pennsylvania, are known to have periodic elevated concentrations of fecal coliform bacteria, a type of bacteria used to indicate the potential presence of fecally related pathogens that may pose health risks to humans exposed through water contact. The availability of near real-time continuous stream discharge, turbidity, and other water-quality data for some streams in the county presents an opportunity to use surrogates to estimate near real-time concentrations of fecal coliform (FC) bacteria and thus provide some information about associated potential health risks during recreational use of streams.The U.S. Geological Survey (USGS), in cooperation with the Chester County Health Department (CCHD) and the Chester County Water Resources Authority (CCWRA), has collected discrete stream samples for analysis of FC concentrations during March–October annually at or near five gaging stations where near real-time continuous data on stream discharge, turbidity, and water temperature have been collected since 2007 (or since 2012 at 2 of the 5 stations). In 2014, the USGS, in cooperation with the CCWRA and CCHD, began to develop regression equations to estimate FC concentrations using available near real-time continuous data. Regression equations included possible explanatory variables of stream discharge, turbidity, water temperature, and seasonal factors calculated using Julian Day with base-10 logarithmic (log) transformations of selected variables.The regression equations were developed using the data from 2007 to 2015 (101–106 discrete bacteria samples per site) for three gaging stations on Brandywine Creek (West Branch Brandywine Creek at Modena, East Branch Brandywine Creek below Downingtown, and Brandywine Creek at Chadds Ford) and from 2012 to 2015 (37–38 discrete bacteria samples per site) for one station each on French Creek near Phoenixville and

  4. Fecal Microbiota Transplantation in Patients With Blood Disorders Inhibits Gut Colonization With Antibiotic-Resistant Bacteria: Results of a Prospective, Single-Center Study.

    PubMed

    Bilinski, Jaroslaw; Grzesiowski, Pawel; Sorensen, Nikolaj; Madry, Krzysztof; Muszynski, Jacek; Robak, Katarzyna; Wroblewska, Marta; Dzieciatkowski, Tomasz; Dulny, Grazyna; Dwilewicz-Trojaczek, Jadwiga; Wiktor-Jedrzejczak, Wieslaw; Basak, Grzegorz W

    2017-08-01

    Patients with blood disorders colonized with antibiotic-resistant bacteria (ARB) are prone to systemic infections that are difficult to treat. Reintroduction of commensal bacteria in a murine model of enterococcal colonization of the gut can lead to eradication of enterococci. We hypothesized that fecal microbiota transplantation (FMT) could be used to eradicate ARB in humans. Participants colonized with ARB were treated with intraduodenal FMT according to a prospective protocol (NCT02461199). The primary endpoint was complete ARB decolonization at 1 month after FMT. Secondary endpoints included safety assessment and partial ARB decolonization. Microbiome sequencing was performed to investigate the influence of microbial composition of the transplanted material on the outcome of FMT. Twenty-five FMTs were performed in 20 participants (including 40% who had neutropenia) who were colonized by a median of 2 (range, 1-4) strains of ARB. The primary endpoint was reached in 15/25 (60%) of the FMTs and more frequently in cases in which there was no periprocedural use of antibiotics (79% vs 36%, P < .05). Among participants, 15/20 (75%) experienced complete ARB decolonization. There were no severe adverse events, and partial ARB decolonization was observed in 20/25 (80%) of the FMTs. The microbiota composition analysis revealed higher abundance of Barnesiella spp., Bacteroides, and Butyricimonas and greater bacterial richness in the fecal material, resulting in eradication of Klebsiella pneumoniae compared with nonresponders. FMT in patients with blood disorders is safe and promotes eradication of ARB from the gastrointestinal tract. NCT02461199.

  5. Bacteroidales Secreted Antimicrobial Proteins Target Surface Molecules Necessary for Gut Colonization and Mediate Competition In Vivo.

    PubMed

    Roelofs, Kevin G; Coyne, Michael J; Gentyala, Rahul R; Chatzidaki-Livanis, Maria; Comstock, Laurie E

    2016-08-23

    . Interference competition, mediated by bacterial factors that directly harm other members, is beginning to be appreciated as important in contributing to species- and strain-level dynamics of abundant gut bacteria. Here, we show that gut Bacteroidales secrete antimicrobial proteins (BSAPs) that antagonize strains of the same species. We show that BSAPs target molecules of sensitive cells that are important for gut colonization and therefore are maintained in sensitive cells. In an experimental animal model of gut colonization, a BSAP-1-producing strain antagonized and outcompeted an isogenic sensitive strain. Furthermore, metagenomic analyses showed that BSAP-1-producing and -sensitive strains are not found together in human gut microbiotas. These data suggest that BSAPs are strong ecological drivers shaping the strain-level composition of gut communities. Copyright © 2016 Roelofs et al.

  6. Mucin degradation in the human colon: production of sialidase, sialate O-acetylesterase, N-acetylneuraminate lyase, arylesterase, and glycosulfatase activities by strains of fecal bacteria.

    PubMed Central

    Corfield, A P; Wagner, S A; Clamp, J R; Kriaris, M S; Hoskins, L C

    1992-01-01

    Oligosaccharide side chains of human colonic mucins contain O-acetylated sialic acids and glycosulfate esters. Although these substituents are considered to protect the chains against degradation by bacterial glycosidases, sialate O-acetylesterase, N-acetylneuraminate lyase, and glycosulfatase activities have been found in fecal extracts. To better define the source of these activities, we measured extracellular and cell-bound sialidase, sialate O-acetylesterase, N-acetylneuraminate lyase, arylesterase, and glycosulfatase activities produced by 23 isolates of human fecal bacteria grown anaerobically in a hog gastric mucin culture medium; these represented dominant populations of fecal anaerobes, facultative anaerobes, and the subset of mucin oligosaccharide-degrading bacteria. Every strain produced sialidase and high levels of arylesterase, and all but five facultative anaerobes produced sialate O-acetylesterase. Sialic acids containing 2 mol or more of O-acetyl ester per mol of sialic acid were cleaved from mucin glycoproteins more slowly by sialidases of mucin oligosaccharide-degrading stains than were sialic acids containing 1 or 0 mol, and only N-acetyl- and mono-O-acetylated sialic acids were recovered from enzyme digests of a mucin containing di-O-acetylated sialic acids. No detectable N-acetylneuraminate lyase activity was produced by any strain, but low activity was induced by increasing the glycoprotein-bound sialic acid concentration in the culture medium of six Escherichia coli strains. Using lactitol-6-sulfate as a substrate, we found weak glycosulfatase activity in the partially purified, concentrated enzyme mixture in the culture supernatants of four mucin oligosaccharide-degrading strains but in none of the unconcentrated culture fractions. We conclude that the presence of two or more O-acetyl groups on sialic acids inhibits enteric bacterial sialidases but that production of sialate O-acetylesterases by several populations of enteric bacteria

  7. Use of DNA Markers for Investigating Sources of Bacteria in Contaminated Ground Water: Wooster Township, Wayne County, Ohio

    USGS Publications Warehouse

    Dumouchelle, Denise H.

    2006-01-01

    In 2004, a public-health nuisance was declared by the Wayne County Board of Health in the Scenic Heights Drive-Batdorf Road area of Wooster Township, Wayne County, Ohio, because of concerns about the safety of water from local wells. Repeated sampling had detected the presence of fecal-indicator bacteria and elevated nitrate concentrations. In June 2006, the U.S. Geological Survey (USGS), in cooperation with the Ohio Environmental Protection Agency (Ohio EPA), collected and analyzed samples from some of the affected wells to help investigate the possibility of human-origin bacterial contamination. Water samples from 12 wells and 5 home sewage-treatment systems (HSTS) were collected. Bromide concentrations were determined in samples from the 12 wells. Samples from 5 of the 12 wells were analyzed for wastewater compounds. Total coliform, enterococci and Escherichia coli (E. coli) bacteria concentrations were determined for samples from 8 of the 12 wells. In addition, two microbial source-tracking tools that employ DNA markers were used on samples from several wells and a composite sample of water from five septic tanks. The DNA markers from the Enterococcus faecium species and the order Bacteroidales are associated with specific sources, either human or ruminant sources. Bromide concentrations ranged from 0.04 to 0.18 milligrams per liter (mg/L). No wastewater compounds were detected at concentrations above the reporting limits. Samples from the 12 wells also were collected by Ohio EPA and analyzed for chloride and nitrate. Chloride concentrations ranged from 12.6 to 61.6 mg/L and nitrate concentrations ranged from 2.34 to 11.9 mg/L (as N). Total coliforms and enterococci were detected in samples from 8 wells, at concentrations from 2 to 200 colony-forming units per 100 milliliters (CFU/100 mL) and 0.5 to 17 CFU/100 mL, respectively. E. coli were detected in samples from three of the eight wells, at concentrations of 1 or 2 CFU/100 mL. Tests for the human

  8. Association of fecal indicator bacteria with human viruses and microbial source tracking markers at coastal beaches impacted by nonpoint source pollution.

    PubMed

    McQuaig, Shannon; Griffith, John; Harwood, Valerie J

    2012-09-01

    Water quality was assessed at two marine beaches in California by measuring the concentrations of culturable fecal indicator bacteria (FIB) and by library-independent microbial source tracking (MST) methods targeting markers of human-associated microbes (human polyomavirus [HPyV] PCR and quantitative PCR, Methanobrevibacter smithii PCR, and Bacteroides sp. strain HF183 PCR) and a human pathogen (adenovirus by nested PCR). FIB levels periodically exceeded regulatory thresholds at Doheny and Avalon Beaches for enterococci (28.5% and 31.7% of samples, respectively) and fecal coliforms (20% and 5.8%, respectively). Adenoviruses were detected at four of five sites at Doheny Beach and were correlated with detection of HPyVs and human Bacteroides HF183; however, adenoviruses were not detected at Avalon Beach. The most frequently detected human source marker at both beaches was Bacteroides HF183, which was detected in 27% of samples. Correlations between FIBs and human markers were much more frequent at Doheny Beach than at Avalon Beach; e.g., adenovirus was correlated with HPyVs and HF183. Human sewage markers and adenoviruses were routinely detected in samples meeting FIB regulatory standards. The toolbox approach of FIB measurement coupled with analysis of several MST markers targeting human pathogens used here demonstrated that human sewage is at least partly responsible for the degradation of water quality, particularly at Doheny Beach, and resulted in a more definitive assessment of recreational water quality and human health risk than reliance on FIB concentrations alone could have provided.

  9. No difference in fecal levels of bacteria or short chain fatty acids in humans, when consuming fruit juice beverages containing fruit fiber, fruit polyphenols, and their combination.

    PubMed

    Wallace, Alison J; Eady, Sarah L; Hunter, Denise C; Skinner, Margot A; Huffman, Lee; Ansell, Juliet; Blatchford, Paul; Wohlers, Mark; Herath, Thanuja D; Hedderley, Duncan; Rosendale, Douglas; Stoklosinski, Halina; McGhie, Tony; Sun-Waterhouse, Dongxiao; Redman, Claire

    2015-01-01

    This study examined the effect of a Boysenberry beverage (750 mg polyphenols), an apple fiber beverage (7.5 g dietary fiber), and a Boysenberry plus apple fiber beverage (750 mg polyphenols plus 7.5 g dietary fiber) on gut health. Twenty-five individuals completed the study. The study was a placebo-controlled crossover study, where every individual consumed 1 of the 4 treatments in turn. Each treatment phase was 4-week long and was followed by a 2-week washout period. The trial beverages were 350 g taken in 2 doses every day (ie, 175 mL taken twice daily). The hypothesis for the study was that the combination of polyphenols and fiber would have a greater benefit on gut health than the placebo product or the fiber or polyphenols on their own. There were no differences in fecal levels of total bacteria, Bacteroides-Prevotella-Porphyromonas group, Bifidobacteriumspecies, Clostridium perfringens, or Lactobacillus species among any of the treatment groups. Fecal short chain fatty acid concentrations did not vary among treatment groups, although prostaglandin E2 concentrations were higher after consumption of the Boysenberry juice beverage. No significant differences were found in quantitative measures of gut health between the Boysenberry juice beverage, the apple fiber beverage, the Boysenberry juice plus apple fiber beverage, and the placebo beverage.

  10. Sediment-water exchange of Vibrio sp. and fecal indicator bacteria: implications for persistence and transport in the Neuse River Estuary, North Carolina, USA.

    PubMed

    Fries, J Stephen; Characklis, Gregory W; Noble, Rachel T

    2008-02-01

    In estuaries, frequent resuspension and deposition of sediment complicate bacterial transport model development by transporting particle-attached bacteria and possibly inducing bacterial responses, such as growth, degradation, or changes in attachment. In order to better characterize these dynamics, observations were made in the Neuse River Estuary (NRE) using the combination of an in situ sampler to monitor the water column and sediment cores to monitor sediment concentrations. Two allochthonous bacteria, Escherichia coli (EC) and Enterococcus sp. (ENT), were selected as proxies for fecal contamination from stormwater runoff. Vibrio sp. (VIB), native to the NRE, was also observed as an autochthonous bacterial group that includes potentially pathogenic species. Two sampling periods were identified as dominated by different suspension types: runoff and resuspension. Despite this difference, several bacterial measures remained comparable between sampling periods. In bottom water, VIB concentration was correlated with salinity and ENT concentration was correlated with turbidity. Differences were observed for EC, where higher concentrations were found in hypoxic waters and sediment during the resuspension period. In the sediment, EC and VIB concentrations significantly increased following the passage of Hurricane Ophelia in September 2005. Throughout this study, all bacterial groups showed evidence of persistence in sediment, suggesting that sediment resuspension represents a significant source of bacteria to the water column.

  11. Are fecal indicator bacteria appropriate measures of recreational water risks in the tropics: A cohort study of beach goers in Brazil?

    PubMed

    Lamparelli, Claudia Condé; Pogreba-Brown, Kristen; Verhougstraete, Marc; Sato, Maria Inês Zanoli; Bruni, Antonio de Castro; Wade, Timothy J; Eisenberg, Joseph N S

    2015-12-15

    Regulating recreational water exposure to pathogens within the tropics is a major public health and economic concern. Although numerous epidemiological studies estimating the risk to recreational marine water exposure have been conducted since the 1950s, few studies have been done in the tropics. Furthermore, many have suggested that the use of fecal indicator bacteria for monitoring recreational water quality in temperate regions is not appropriate in the tropics. We analyzed a large cohort study of five beaches in Sao Paulo, Brazil, conducted during consecutive weekends in the summer of 1999 that estimated risk to water, sand, and food exposures. Enterococci and Escherichia coli concentrations were measured each day of the study. Elevated risks were estimated for both swimming (OR = 1.36 95% CI: 1.05-1.58) and sand contact (OR = 1.29 95% CI 1.05-1.58). A 1 log increase in enterococci concentration was associated with an 11% increase in risk (OR = 1.11 95% CI: 1.04-1.19). For E. coli a 1-log increase in concentration was associated with 19% increase in risk (OR = 1.19 95% CI: 1.14-1.28). Most countries with beaches in the tropics are lower or middle income countries (LMIC) and rely on tourism as a major source of income. We present data that suggests fecal indicator bacteria such as enterococci are an appropriate indicator of risk in tropical urban settings where contamination is coming from predominantly human sources. Additional studies in tropical settings could help inform and refine guidelines for safe use of recreational waters.

  12. Effects of Holding Time, Storage, and the Preservation of Samples on Sample Integrity for the Detection of Fecal Indicator Bacteria by Quantitative Polymerase Chain Reaction (qPCR)-based assays.

    EPA Science Inventory

    The purpose of this project was to answer questions related to storage of samples to be analyzed by the quantitative polymerase chain reaction (qPCR)-based assays for fecal indicator bacteria. The project was divided into two parts. The first part was to determine if filters th...

  13. The Relationship Between Land Management, Fecal Indicator Bacteria, and the Occurrence of Campylobacter and Listeria Spp. in Water and Sediments During Synoptic Sampling In The South Fork Broad River Watershed, Northeast Georgia, U.S.A.

    EPA Science Inventory

    Fecal indicator bacteria (FIB) and pathogens stored in the bed sediments of streams and rivers may be mobilized into the water column affecting overall water quality. Furthermore, land management may play an important role in the concentrations of FIB and the occurrence of pathog...

  14. Effects of Holding Time, Storage, and the Preservation of Samples on Sample Integrity for the Detection of Fecal Indicator Bacteria by Quantitative Polymerase Chain Reaction (qPCR)-based assays.

    EPA Science Inventory

    The purpose of this project was to answer questions related to storage of samples to be analyzed by the quantitative polymerase chain reaction (qPCR)-based assays for fecal indicator bacteria. The project was divided into two parts. The first part was to determine if filters th...

  15. The Relationship Between Land Management, Fecal Indicator Bacteria, and the Occurrence of Campylobacter and Listeria Spp. in Water and Sediments During Synoptic Sampling In The South Fork Broad River Watershed, Northeast Georgia, U.S.A.

    EPA Science Inventory

    Fecal indicator bacteria (FIB) and pathogens stored in the bed sediments of streams and rivers may be mobilized into the water column affecting overall water quality. Furthermore, land management may play an important role in the concentrations of FIB and the occurrence of pathog...

  16. Characterization of sources and loadings of fecal pollutants using microbial source tracking assays in urban and rural areas of the Grand River Watershed, Southwestern Ontario.

    PubMed

    Lee, Dae-Young; Lee, Hung; Trevors, Jack T; Weir, Susan C; Thomas, Janis L; Habash, Marc

    2014-04-15

    Sources of fecal water pollution were assessed in the Grand River and two of its tributaries (Ontario, Canada) using total and host-specific (human and bovine) Bacteroidales genetic markers in conjunction with reference information, such as land use and weather. In-stream levels of the markers and culturable Escherichia coli were also monitored during multiple rain events to gain information on fecal loadings to catchment from diffuse sources. Elevated human-specific marker levels were accurately identified in river water impacted by a municipal wastewater treatment plant (WWTP) effluent and at a downstream site in the Grand River. In contrast, the bovine-specific marker showed high levels of cattle fecal pollution in two tributaries, both of which are characterized as intensely farmed areas. The bovine-specific Bacteroidales marker increased with rainfall in the agricultural tributaries, indicating enhanced loading of cattle-derived fecal pollutants to river from non-point sources following rain events. However, rain-triggered fecal loading was not substantiated in urban settings, indicating continuous inputs of human-originated fecal pollutants from point sources, such as WWTP effluent. This study demonstrated that the Bacteroidales source tracking assays, in combination with land use information and hydrological data, may provide additional insight into the spatial and temporal distribution of source-specific fecal contamination in streams impacted by varying land uses. Using the approach described in this study may help to characterize impacted water sources and to design targeted land use management plans in other watersheds in the future.

  17. Decay of Bacterial Pathogens, Fecal Indicators, and Real-Time Quantitative PCR Genetic Markers in Manure-Amended Soils ▿ †

    PubMed Central

    Rogers, Shane W.; Donnelly, Matthew; Peed, Lindsay; Kelty, Catherine A.; Mondal, Sumona; Zhong, Zirong; Shanks, Orin C.

    2011-01-01

    This study examined persistence and decay of bacterial pathogens, fecal indicator bacteria (FIB), and emerging real-time quantitative PCR (qPCR) genetic markers for rapid detection of fecal pollution in manure-amended agricultural soils. Known concentrations of transformed green fluorescent protein-expressing Escherichia coli O157:H7/pZs and red fluorescent protein-expressing Salmonella enterica serovar Typhimurium/pDs were added to laboratory-scale manure-amended soil microcosms with moisture contents of 60% or 80% field capacity and incubated at temperatures of −20°C, 10°C, or 25°C for 120 days. A two-stage first-order decay model was used to determine stage 1 and stage 2 first-order decay rate coefficients and transition times for each organism and qPCR genetic marker in each treatment. Genetic markers for FIB (Enterococcus spp., E. coli, and Bacteroidales) exhibited decay rate coefficients similar to that of E. coli O157:H7/pZs but not of S. enterica serovar Typhimurium/pDs and persisted at detectable levels longer than both pathogens. Concentrations of these two bacterial pathogens, their counterpart qPCR genetic markers (stx1 and ttrRSBCA, respectively), and FIB genetic markers were also correlated (r = 0.528 to 0.745). This suggests that these qPCR genetic markers may be reliable conservative surrogates for monitoring fecal pollution from manure-amended land. Host-associated qPCR genetic markers for microbial source tracking decayed rapidly to nondetectable concentrations, long before FIB, Salmonella enterica serovar Typhimurium/pDs, and E. coli O157:H7/pZs. Although good indicators of point source or recent nonpoint source fecal contamination events, these host-associated qPCR genetic markers may not be reliable indicators of nonpoint source fecal contamination events that occur weeks following manure application on land. PMID:21642395

  18. Effect of dietary inorganic sulfur level on growth performance, fecal composition, and measures of inflammation and sulfate-reducing bacteria in the intestine of growing pigs.

    PubMed

    Kerr, B J; Weber, T E; Ziemer, C J; Spence, C; Cotta, M A; Whitehead, T R

    2011-02-01

    Two experiments investigated the impact of dietary inorganic S on growth performance, intestinal inflammation, fecal composition, and the presence of sulfate-reducing bacteria (SRB). In Exp. 1, individually housed pigs (n = 42; 13.8 kg) were fed diets containing 2,300 or 2,100 mg/kg of S for 24 d. Decreasing dietary S had no effect on ADG, ADFI, or G:F. In Exp. 2, pigs (n = 64; 13.3 kg) were fed diets containing 0, 0.625, 1.25, 2.5, or 5.0% CaSO(4), thereby increasing dietary S from 2,900 to 12,100 mg/kg. Two additional diets were fed to confirm the lack of an impact due to feeding low dietary S on pig performance and to determine if the increased Ca and P content in the diets containing CaSO(4) had an impact on growth performance. Pigs were fed for 35 d. Ileal tissue, ileal mucosa, and colon tissue were harvested from pigs fed the 0 and 5% CaSO(4) diets (low-S and high-S, respectively) to determine the impact of dietary S on inflammation-related mRNA, activity of mucosal alkaline phosphatase and sucrase, and pathways of inflammatory activation. Real-time PCR was used to quantify SRB in ileal and colon digesta samples and feces. Fecal pH, sulfide, and ammonia concentrations were also determined. There was no impact on growth performance in pigs fed the diet reduced in dietary S or by the increase of dietary Ca and P. Increasing dietary S from 2,900 to 12,100 mg/kg had a linear (P < 0.01) effect on ADG and a cubic effect (P < 0.05) on ADFI and G:F. Real-time reverse-transcription PCR analysis revealed that pigs fed high-S increased (P < 0.05) the relative abundance of intracellular adhesion molecule-1, tumor necrosis factor-α, and suppressor of cytokine signaling-3 mRNA, and tended (P = 0.09) to increase the relative abundance of IL-6 mRNA in ileal tissue. Likewise, pigs fed high-S had reduced (P < 0.05) abundance of nuclear factor of κ light polypeptide gene enhancer in B-cells inhibitor-α and increased (P < 0.05) phospho-p44/p42 mitogen-activated protein kinase

  19. Occurrence and levels of fecal indicators and pathogenic bacteria in market-ready recycled organic matter composts.

    PubMed

    Brinton, W F; Storms, P; Blewett, T C

    2009-02-01

    Landfill diversion of organic wastes through composting is making compost products available for agricultural and horticultural crops. On certified organic farms, nonsludge green waste and manure composts are widely used because the use of these products removes harvest date restrictions imposed by the U.S. Department of Agriculture when raw manure is applied. We quantified several pathogens in point-of-sale composts from 94 nonsludge facilities processing 2.2 million m3 year(-1) of recycled green waste. Only one compost contained Salmonella (1.8 most probable number [MPN]/4 g), 28% had fecal coliforms exceeding the Environmental Protection Agency 503 sludge hygiene limits (1000 MPN g(-1)), and 6% had detectable Escherichia coli O157:H7. In 22 of 47 samples, very low levels of Listeria spp. were found. However, in one sample the Listeria level was very high, coinciding with the highest overall level of all pathogen indicators. Seventy percent of the compost samples were positive for Clostridium perfringens, but only 20% of the samples had levels >1000 CFU/g. All samples were positive for fecal streptococci, and 47% had >1000 MPN g(-1). Statistical analyses conducted using documented site characteristics revealed that factors contributing to elevated pathogen levels were large facility size, large pile size, and immaturity of compost. Application of the California Compost Maturity Index distinguished compost products that had very low levels of E. coli from those with high levels. Products produced with windrow methods were of higher microbiological quality than were those produced with static pile methods, and point-of-sale bagged composts scored very high. These data indicate that compost that is hygienic by common standards can be produced, but more effort is required to improve hygiene consistency in relation to management practices.

  20. Changes in antimicrobial resistance in fecal bacteria associated with pig transit and holding times at slaughter plants.

    PubMed Central

    Molitoris, E; Fagerberg, D J; Quarles, C L; Krichevsky, M I

    1987-01-01

    Fecal coliforms, fecal streptococci, and antimicrobial resistance (AMR) associated with various pig transit and holding times were investigated at slaughter plants. Changes in the relative abundance of two biotypes of Streptococcus faecium were associated with transit and holding of pigs, although approximately 20% of the isolates were unidentified. The greatest variety of coliforms was isolated from porcine feces after short transit (2 h) or holding (3 h) times and was qualitatively similar to those from pigs on farms. Isolates from pigs with longer average transit or holding times were almost all Escherichia coli (four biotypes). Streptococcal resistance to most antimicrobial agents was significantly greater (P less than 0.05) in isolates from live pigs at slaughter plants than in those from pigs at farms and was apparent after a short transit time (2 h). Streptococci from pigs held an average of 15 h were less resistant to most antimicrobial agents than those from pigs held 3 or 43 h. When compared with short transit times, moderate transit times (6 h) were associated with significantly decreased (P less than 0.05) coliform resistance and decreased resistance transfer but a greater diversity of AMR patterns. Holding pigs overnight (14 h) was associated with lowered coliform resistance to several antimicrobial agents, compared with the resistance of isolates from pigs held 3 or 39 h. A substantial increase (18 to 48%) in the ability to transfer streptomycin resistance was demonstrated in coliforms from pigs held 39 h, when compared with those from pigs held 3 h.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:3606107

  1. Photoelectrocatalytic inactivation of fecal coliform bacteria in urban wastewater using nanoparticulated films of TiO2 and TiO2/Ag.

    PubMed

    Domínguez-Espíndola, Ruth Belinda; Varia, Jeet C; Álvarez-Gallegos, Alberto; Ortiz-Hernández, Ma Laura; Peña-Camacho, Justina Leticia; Silva-Martínez, Susana

    2017-03-01

    Photocatalysis has shown the ability to inactivate a wide range of harmful microorganisms with traditional use of chlorination. Photocatalysis combined with applied bias potential (photoelectrocatalysis) increases the efficiency of photocatalysis and decreases the charge recombination. This work examines the inactivation of fecal coliform bacteria present in real urban wastewater by photoelectrocatalysis using nanoparticulated films of TiO2 and TiO2/Ag (4%w/w) under UV light irradiation. The catalysts were prepared with different thicknesses by the sol-gel method and calcined at 400°C and 600°C. The urban wastewater samples were collected from the sedimentation tank effluent of the university sewage treatment facility. The rate of bacteria inactivation increases with increasing the applied potential and film thicknesses; also, the presence of silver on the catalyst surface annealed at 400°C shows better inactivation than that at 600°C. Finally, a structural cell damage of Escherichia coli (DH5α), inoculated in water, is observed during the photoelectrocatalytic process.

  2. Improved real-time PCR assays for the detection of fecal indicator bacteria in surface waters with different instrument and reagent systems.

    PubMed

    Siefring, S; Varma, M; Atikovic, E; Wymer, L; Haugland, R A

    2008-06-01

    Previously reported and redesigned primer and probe assays were evaluated for the quantitative analysis of the fecal indicator bacterial groups, Enterococcus and Bacteroidetes with three real-time PCR instrument and reagent systems. The efficiency and sensitivity of the original assays varied between systems in analyses of DNA extracts from pure cultures of Enterococcus faecalis and Bacteroides fragilis, whereas the modified assays gave more consistent results. Distinctions between original and modified assays also occurred in analyses of known spike levels of E. faecalis and B. fragilis cells on filters with diverse surface water retentates. Percentages of samples causing PCR failures due to inhibition were lower using the modified assays. The accuracy and precision of spiked bacteria measurements were also generally higher, although mean measurements of both target organisms were still significantly different between systems (p < 0.05). The accuracy and precision of spiked bacteria measurements by both modified assays were further improved using a new sample matrix control spike consisting of cultured Lactococcus lactis cells and a reference assay for this organism. Corrections provided by the L. lactis assay eliminated significant differences in E. faecalis measurements between all three systems and between two of the three systems in B. fragilis measurements.

  3. Predominant genera of fecal microbiota in children with atopic dermatitis are not altered by intake of probiotic bacteria Lactobacillus acidophilus NCFM and Bifidobacterium animalis subsp. lactis Bi-07.

    PubMed

    Larsen, Nadja; Vogensen, Finn K; Gøbel, Rikke; Michaelsen, Kim F; Abu Al-Soud, Waleed; Sørensen, Søren J; Hansen, Lars H; Jakobsen, Mogens

    2011-03-01

    The effect of probiotic bacteria Lactobacillus acidophilus NCFM and Bifidobacterium lactis Bi-07 on the composition of the Lactobacillus group, Bifidobacterium and the total bacterial population in feces from young children with atopic dermatitis was investigated. The study included 50 children randomized to intake of one of the probiotic strain or placebo. Microbial composition was characterized by denaturing gradient gel electrophoresis, quantitative PCR and, in a subset of subjects, by pyrosequencing of the 16S rRNA gene. The core population of the Lactobacillus group was identified as Lactobacillus gasseri, Lactobacillus fermentum, Lactobacillus oris, Leuconostoc mesenteroides, while the bifidobacterial community included Bifidobacterium adolescentis, Bifidobacterium bifidum, Bifidobacterium longum and Bifidobacterium catenulatum. The fecal numbers of L. acidophilus and B. lactis increased significantly after intervention, indicating survival of the ingested bacteria. The levels of Bifidobacterium correlated positively (P=0.03), while the levels of the Lactobacillus group negatively (P=0.01) with improvement of atopic eczema evaluated by the Severity Scoring of Atopic Dermatitis index. This correlation was observed across the whole study cohort and not attributed to the probiotic intake. The main conclusion of the study is that administration of L. acidophilus NCFM and B. lactis Bi-07 does not affect the composition and diversity of the main bacterial populations in feces. © 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

  4. Use of salinity mixing models to estimate the contribution of creek water fecal indicator bacteria to an estuarine environment: Newport Bay, California.

    PubMed

    McLaughlin, Karen; Ahn, Jong Ho; Litton, Rachel M; Grant, Stanley B

    2007-08-01

    The contribution of freshwater discharge to fecal indicator bacteria (FIB) impairment of an estuarine environment can be approximated from simple, two end-member mixing models using salinity as a tracer. We conducted a yearlong time series investigation of Newport Bay, a regionally important estuarine embayment in southern California, assessing the concentrations of FIB, specifically Escherichia coli and enterococci bacteria, and salinity. In total, eight within-bay stations and one offshore control site were sampled nearly once per week and the three tributaries draining into Newport Bay were sampled approximately daily. Using salinity as a conservative tracer for water mass mixing and determining the end-member values of FIB in both the creek sites and the offshore site, we created a linear, two end-member mixing model of FIB within Newport Bay. Deviations from the mixing model suggest either an additional source of FIB to the bay (e.g. bird feces, storm drain discharge) or regrowth and/or die-off of FIB within the bay. Our results indicate that salinity mixing models can be useful in predicting changes in FIB concentrations in the estuarine environments and can help narrow the search for sources of FIB to the bay and enhance our understanding of the fate of FIB within the bay.

  5. Incorporating expert judgments in utility evaluation of bacteroidales qPCR assays for microbial source tracking in a drinking water source.

    PubMed

    Åström, Johan; Pettersson, Thomas J R; Reischer, Georg H; Norberg, Tommy; Hermansson, Malte

    2015-02-03

    Several assays for the detection of host-specific genetic markers of the order Bacteroidales have been developed and used for microbial source tracking (MST) in environmental waters. It is recognized that the source-sensitivity and source-specificity are unknown and variable when introducing these assays in new geographic regions, which reduces their reliability and use. A Bayesian approach was developed to incorporate expert judgments with regional assay sensitivity and specificity assessments in a utility evaluation of a human and a ruminant-specific qPCR assay for MST in a drinking water source. Water samples from Lake Rådasjön were analyzed for E. coli, intestinal enterococci and somatic coliphages through cultivation and for human (BacH) and ruminant-specific (BacR) markers through qPCR assays. Expert judgments were collected regarding the probability of human and ruminant fecal contamination based on fecal indicator organism data and subjective information. Using Bayes formula, the conditional probability of a true human or ruminant fecal contamination given the presence of BacH or BacR was determined stochastically from expert judgments and regional qPCR assay performance, using Beta distributions to represent uncertainties. A web-based computational tool was developed for the procedure, which provides a measure of confidence to findings of host-specific markers and demonstrates the information value from these assays.

  6. Incorporating Expert Judgments in Utility Evaluation of Bacteroidales qPCR Assays for Microbial Source Tracking in a Drinking Water Source

    PubMed Central

    Åström, Johan; Pettersson, Thomas J. R.; Reischer, Georg H.; Norberg, Tommy; Hermansson, Malte

    2017-01-01

    Several assays for the detection of host-specific genetic markers of the order Bacteroidales have been developed and used for microbial source tracking (MST) in environmental waters. It is recognized that the source-sensitivity and source-specificity are unknown and variable when introducing these assays in new geographic regions, which reduces their reliability and use. A Bayesian approach was developed to incorporate expert judgments with regional assay sensitivity and specificity assessments in a utility evaluation of a human and a ruminant-specific qPCR assay for MST in a drinking water source. Water samples from Lake Rådasjön were analyzed for E. coli, intestinal enterococci and somatic coliphages through cultivation and for human (BacH) and ruminant-specific (BacR) markers through qPCR assays. Expert judgments were collected regarding the probability of human and ruminant fecal contamination based on fecal indicator organism data and subjective information. Using Bayes formula, the conditional probability of a true human or ruminant fecal contamination given the presence of BacH or BacR was determined stochastically from expert judgments and regional qPCR assay performance, using Beta distributions to represent uncertainties. A web-based computational tool was developed for the procedure, which provides a measure of confidence to findings of host-specific markers and demonstrates the information value from these assays. PMID:25545113

  7. Runoff delay exerts a strong control on the field-scale removal of manure-borne fecal bacteria with runoff

    USDA-ARS?s Scientific Manuscript database

    The microbial safety of surface waters is an ongoing issue which is threatened by the transport of manure-borne bacteria to water sources used for irrigation or recreation. Predictive modeling has become an effective tool to forecast the microbial quality of water during precipitation events, howeve...

  8. Runoff delay exerts a strong control on the field-scale removal of manure-borne fecal bacteria with runoff

    USDA-ARS?s Scientific Manuscript database

    The microbial safety of surface waters is an ongoing issue which is threatened by the transport of manure-borne bacteria to water sources used for irrigation or recreation. Predictive modeling has become an effective tool to forecast the microbial quality of water duringprecipitation events, however...

  9. Comparison of the multiple-sample means with composite sample results for fecal indicator bacteria by quantitative PCR and culture.

    PubMed

    Converse, Reagan R; Wymer, Larry J; Dufour, Alfred P; Wade, Timothy J

    2012-10-01

    Few studies have addressed the efficacy of composite sampling for measuring indicator bacteria by quantitative PCR (qPCR). We compared results from composited samples with multiple-sample means for culture- and qPCR-based water quality monitoring. Results from composited samples for both methods were similarly correlated to multiple-sample means and predicted criteria exceedances equally.

  10. New submodel for watershed-scale simulations of fecal bacteria fate and transport at agricultural and pasture lands

    USDA-ARS?s Scientific Manuscript database

    Microbial contamination of waters is the critical public health issue. The watershed-scale process-based modeling of bacteria fate and transport (F&T) has been proven to serve as the useful tool for predicting microbial water quality and evaluating management practices. The objective of this work is...

  11. Comparison of the Multiple-sample means with composite sample results for fecal indicator bacteria by quantitative PCR and culture

    EPA Science Inventory

    ABSTRACT: Few studies have addressed the efficacy of composite sampling for measurement of indicator bacteria by QPCR. In this study, composite results were compared to single sample results for culture- and QPCR-based water quality monitoring. Composite results for both methods ...

  12. Comparison of the Multiple-sample means with composite sample results for fecal indicator bacteria by quantitative PCR and culture

    EPA Science Inventory

    ABSTRACT: Few studies have addressed the efficacy of composite sampling for measurement of indicator bacteria by QPCR. In this study, composite results were compared to single sample results for culture- and QPCR-based water quality monitoring. Composite results for both methods ...

  13. Survival dynamics of fecal bacteria in ponds in agricultural watersheds of the Piedmont and Coastal Plain of Georgia

    USDA-ARS?s Scientific Manuscript database

    Animal agriculture in watersheds can be a source of manure bacteria that can contaminate surface waters and put public health at risk. Because of the expanding urban-agriculture interface preventing surface water contamination with manure pathogens is important for sustaining surface water quality. ...

  14. Characterizing relationships among fecal indicator bacteria, microbial source tracking markers, and associated waterborne pathogen occurrence in stream water and sediments in a mixed land use watershed.

    PubMed

    Bradshaw, J Kenneth; Snyder, Blake J; Oladeinde, Adelumola; Spidle, David; Berrang, Mark E; Meinersmann, Richard J; Oakley, Brian; Sidle, Roy C; Sullivan, Kathleen; Molina, Marirosa

    2016-09-15

    Bed sediments of streams and rivers may store high concentrations of fecal indicator bacteria (FIB) and pathogens. Due to resuspension events, these contaminants can be mobilized into the water column and affect overall water quality. Other bacterial indicators such as microbial source tracking (MST) markers, developed to determine potential sources of fecal contamination, can also be resuspended from bed sediments. The primary objective of this study was to predict occurrence of waterborne pathogens in water and streambed sediments using a simple statistical model that includes traditionally measured FIB, environmental parameters and source allocation, using MST markers as predictor variables. Synoptic sampling events were conducted during baseflow conditions downstream from agricultural (AG), forested (FORS), and wastewater pollution control plant (WPCP) land uses. Concentrations of FIB and MST markers were measured in water and sediments, along with occurrences of the enteric pathogens Campylobacter, Listeria and Salmonella, and the virulence gene that carries Shiga toxin, stx2. Pathogens were detected in water more often than in underlying sediments. Shiga toxin was significantly related to land use, with concentrations of the ruminant marker selected as an independent variable that could correctly classify 76% and 64% of observed Shiga toxin occurrences in water and sediment, respectively. FIB concentrations and water quality parameters were also selected as independent variables that correctly classified Shiga toxin occurrences in water and sediment (54%-87%), and Salmonella occurrences in water (96%). Relationships between pathogens and indicator variables were generally inconsistent and no single indicator adequately described occurrence of all pathogens. Because of inconsistent relationships between individual pathogens and FIB/MST markers, incorporating a combination of FIB, water quality measurements, and MST markers may be the best way to assess

  15. Marsh soils as potential sinks for Bacteroides fecal indicator bacteria, Waccamaw National Wildlife Refuge, Georgetown, SC, USA

    USGS Publications Warehouse

    Drexler, Judith Z.; Johnson, Heather E.; Duris, Joseph W.; Krauss, Ken W.

    2014-01-01

    A soil core collected in a tidal freshwater marsh in the Waccamaw National Wildlife Refuge (Georgetown, SC) exuded a particularly strong odor of cow manure upon extrusion. In order to test for manure and determine its provenance, we carried out microbial source tracking using DNA markers for Bacteroides, a noncoliform, anaerobic bacterial group that represents a broad group of the fecal population. Three core sections from 0-3 cm, 9-12 cm and 30-33 were analyzed for the presence of Bacteroides. The ages of core sediments were estimated using 210Pb and 137Cs dating. All three core sections tested positive for Bacteroides DNA markers related to cow or deer feces. Because cow manure is stockpiled, used as fertilizer, and a source of direct contamination in the Great Pee Dee River/Winyah Bay watershed, it is very likely the source of the Bacteroides that was deposited on the marsh. The mid-points of the core sections were dated as follows: 0-3 cm: 2009; 9-12 cm: 1999, and 30-33 cm: 1961. The presence of Bacteroides at different depths/ages in the soil profile indicates that soils in tidal freshwater marshes are, at the least, capable of being short-term sinks for Bacteroides and, may have the potential to be long-term sinks of stable, naturalized populations.

  16. Discovering new indicators of fecal pollution

    PubMed Central

    McLellan, Sandra L.; Eren, A. Murat

    2014-01-01

    Fecal pollution indicators are essential to identify and remediate contamination sources and protect public health. Historically, easily cultured facultative anaerobes such as fecal coliforms, Escherichia coli, or enterococci have been used, but these indicators generally provide no information as to their source. More recently, molecular methods have targeted fecal anaerobes, which are much more abundant in humans and other mammals and some strains appear to be associated with certain host sources. Next-generation sequencing and microbiome studies have created an unprecedented inventory of microbial communities associated with fecal sources, allowing reexamination of which taxonomic groups are best suited as informative indicators. The use of new computational methods, such as oligotyping coupled with well-established machine learning approaches, is providing new insights into patterns of host association. In this review we examine the basis for host-specificity and the rationale for using 16S rRNA gene targets for alternative indicators and highlight two taxonomic groups, Bacteroidales and Lachnospiraceae, which are rich in host-specific bacterial organisms. Finally, we discuss considerations for using alternative indicators for water quality assessments with a particular focus on detecting human sewage sources of contamination. PMID:25199597

  17. Diversity and distribution of commensal fecal Escherichia coli bacteria in beef cattle administered selected subtherapeutic antimicrobials in a feedlot setting.

    PubMed

    Sharma, Ranjana; Munns, Krysty; Alexander, Trevor; Entz, Toby; Mirzaagha, Parasto; Yanke, L Jay; Mulvey, Michael; Topp, Edward; McAllister, Tim

    2008-10-01

    Escherichia coli strains isolated from fecal samples were screened to examine changes in phenotypic and genotypic characteristics including antimicrobial susceptibility, clonal type, and carriage of resistance determinants. The goal of this 197-day study was to investigate the influence of administration of chlortetracycline alone (T) or in combination with sulfamethazine (TS) on the development of resistance, dissemination of defined strain types, and prevalence of resistance determinants in feedlot cattle. Inherent tetracycline resistance was detected in cattle with no prior antimicrobial exposure. Antimicrobial administration was not found to be essential for the maintenance of inherently ampicillin-resistant and tetracycline-resistant (Tet(r)) E. coli in control animals; however, higher Tet(r) E. coli shedding was observed in animals subjected to the two treatments. At day 0, high tetracycline (26.7%), lower sulfamethoxazole-tetracycline (19.2%), and several other resistances were detected, which by the finishing phase (day 197) were restricted to ampicillin-tetracycline (47.5%), tetracycline (31.7%), and ampicillin-tetracycline-sulfamethoxazole (20.8%) from both treated and untreated cattle. Among the determinants, bla(TEM1), tet(A), and sul2 were prevalent at days 0 and 197. Further, E. coli from day 0 showed diverse antibiogram profiles and strain types, which by the finishing phase were limited to up to three, irrespective of the treatment. Some genetically identical strains expressed different phenotypes and harbored diverse determinants, indicating that mobile genetic elements contribute to resistance dissemination. This was supported by an increased linked inheritance of ampicillin and tetracycline resistance genes and prevalence of specific strains at day 197. Animals in the cohort shed increasingly similar genotypes by the finishing phase due to animal-to-animal strain transmission. Thus, characterizing inherent resistance and propagation of cohort

  18. Occurrence of dissolved solids, nutrients, atrazine, and fecal coliform bacteria during low flow in the Cheney Reservoir watershed, south-central Kansas, 1996

    USGS Publications Warehouse

    Christensen, V.G.; Pope, L.M.

    1997-01-01

    A network of 34 stream sampling sites was established in the 1,005-square-mile Cheney Reservoir watershed, south-central Kansas, to evaluate spatial variability in concentrations of selected water-quality constituents during low flow. Land use in the Cheney Reservoir watershed is almost entirely agricultural, consisting of pasture and cropland. Cheney Reservoir provides 40 to 60 percent of the water needs for the city of Wichita, Kansas. Sampling sites were selected to determine the relative contribution of point and nonpoint sources of water-quality constituents to streams in the watershed and to identify areas of potential water-quality concern. Water-quality constituents of interest included dissolved solids and major ions, nitrogen and phosphorus nutrients, atrazine, and fecal coliform bacteria. Water from the 34 sampling sites was sampled once in June and once in September 1996 during Phase I of a two-phase study to evaluate water-quality constituent concentrations and loading characteristics in selected subbasins within the watershed and into and out of Cheney Reservoir. Information summarized in this report pertains to Phase I and was used in the selection of six long-term monitoring sites for Phase II of the study. The average low-flow constituent concentrations in water collected during Phase I from all sampling sites was 671 milligrams per liter for dissolved solids, 0.09 milligram per liter for dissolved ammonia as nitrogen, 0.85 milligram per liter for dissolved nitrite plus nitrate as nitrogen, 0.19 milligram per liter for total phosphorus, 0.20 microgram per liter for dissolved atrazine, and 543 colonies per 100 milliliters of water for fecal coliform bacteria. Generally, these constituents were of nonpoint-source origin and, with the exception of dissolved solids, probably were related to agricultural activities. Dissolved solids probably occur naturally as the result of the dissolution of rocks and ancient marine sediments containing large salt

  19. Antimicrobial resistance characteristics and fitness of Gram-negative fecal bacteria from volunteers treated with minocycline or amoxicillin

    PubMed Central

    Kirchner, Miranda; Mafura, Muriel; Hunt, Theresa; Abu-Oun, Manal; Nunez-Garcia, Javier; Hu, Yanmin; Weile, Jan; Coates, Anthony; Card, Roderick; Anjum, Muna F.

    2014-01-01

    A yearlong study was performed to examine the effect of antibiotic administration on the bacterial gut flora. Gram-negative facultative anaerobic bacteria were recovered from the feces of healthy adult volunteers administered amoxicillin, minocycline or placebo, and changes determined in antimicrobial resistance (AMR) gene carriage. Seventy percent of the 1039 facultative anaerobic isolates recovered were identified by MALDI-TOF as Escherichia coli. A microarray used to determine virulence and resistance gene carriage demonstrated that AMR genes were widespread in all administration groups, with the most common resistance genes being blaTEM, dfr, strB, tet(A), and tet(B). Following amoxicillin administration, an increase in the proportion of amoxicillin resistant E. coli and a three-fold increase in the levels of blaTEM gene carriage was observed, an effect not observed in the other two treatment groups. Detection of virulence genes, including stx1A, indicated not all E. coli were innocuous commensals. Approximately 150 E. coli collected from 6 participants were selected for pulse field gel electrophoresis (PFGE), and a subset used for characterisation of plasmids and Phenotypic Microarrays (PM). PFGE indicated some E. coli clones had persisted in volunteers for up to 1 year, while others were transient. Although there were no unique characteristics associated with plasmids from persistent or transient isolates, PM assays showed transient isolates had greater adaptability to a range of antiseptic biocides and tetracycline; characteristics which were lost in some, but not all persistent isolates. This study indicates healthy individuals carry bacteria harboring resistance to a variety of antibiotics and biocides in their intestinal tract. Antibiotic administration can have a temporary effect of selecting bacteria, showing co-resistance to multiple antibiotics, some of which can persist within the gut for up to 1 year. PMID:25566232

  20. Tracking the Sources of Fecal Contaminations: an Interdisciplinary Toolbox

    NASA Astrophysics Data System (ADS)

    Jeanneau, L.; Jarde, E.; Derrien, M.; Gruau, G.; Solecki, O.; Pourcher, A.; Marti, R.; Wéry, N.; Caprais, M.; Gourmelon, M.; Mieszkin, S.; Jadas-Hécart, A.; Communal, P.

    2011-12-01

    Fecal contaminations of inland and coastal waters induce risks to human health and economic losses. In order to improve water management, it is necessary to identify the sources of contamination, which implies the development of specific markers. In order to be considered as a valuable host-specific marker, one must (1) be source specific, (2) occur in high concentration in polluting matrices, (3) exhibit extra-intestinal persistence similar to fecal indicator bacteria (FIB) and (4) not grow out of the host. However, up to day no single marker has fulfilled all those criteria. Thus, it has been suggested to use a combination of markers in order to generate more reliable data. This has lead to the development of a Microbial Source Tracking (MST) toolbox including FIB and microbial and chemical specific markers in order to differentiate between human, bovine and porcine fecal contaminations. Those specific markers are, (1) genotypes of F-specific RNA bacteriophages, (2) bacterial markers belonging to the Bacteroidales (human-specific HF183, ruminant-specific Rum-2-Bac and pig-specific Pig-2-Bac markers), to the Bifidobacterium (Bifidobacterium adolescentis) and pig-specific Lactobacillus amylovorus, (3) fecal stanols and (4) caffeine. The development of this MST toolbox was composed of four steps, from the molecular scale to the watershed scale. At the molecular scale, the specificity and the concentration of those markers were studied in cattle and pig manures and in waste water treatment plant (WWTP) effluents and influents. At the microcosm scale, the transfer of bovine and porcine specific markers was investigated by rainfall simulations on agricultural plots amended with cattle or pig manure. Moreover, the relative persistence of FIB and human, porcine and bovine specific markers was investigated in freshwater and seawater microcosms inoculated with a WWTP influent, pig manure and cow manure. Finally, the aforementioned MST toolbox has been validated at the

  1. Survival and persistence of human and ruminant-specific faecal Bacteroidales in freshwater microcosms.

    PubMed

    Walters, Sarah P; Field, Katharine G

    2009-06-01

    Amplification of host-specific markers from Bacteroidales faecal anaerobes can rapidly identify the source of faecal pollution. It is necessary to understand persistence and survival of these markers and marker cells, both to interpret quantitative source-tracking data, and to use such data to predict pathogen occurrence. We measured marker persistence and cell survival of two human (HF134, HF183) and two ruminant (CF128, CF193) faecal Bacteroidales markers, compared with Escherichia coli and enterococci. Freshwater microcosms were inoculated with fresh cattle or human faeces and incubated at 13 degrees C in natural light or darkness. Marker persistence was measured by polymerase chain reaction (PCR) and quantitative PCR. Survival of marker cells was measured by real-time quantitative PCR. There was no difference in persistence between the two human-specific Bacteroidales DNA markers in the light and dark microcosms. Cell survival profiles of the two human markers were also similar; both were significantly affected by light. Ruminant markers persisted and survived longer than human markers (14 versus 6 days respectively). CF193 decreased more rapidly than CF128, and light significantly affected CF128 but not CF193. These results support use of host-specific faecal Bacteroidales markers as indicators of recent faecal pollution, but suggest that caution is needed in interpreting quantitative results to indicate proportional contribution of different sources, as individual markers differ in their survival, persistence and response to environmental variables. The survival and persistence profiles for Bacteroidales markers are consistent with survival profiles for several faecal pathogens.

  2. An assessment of fecal indicator and other bacteria from an urbanized coastal lagoon in the City of Los Angeles, California, USA.

    PubMed

    Dorsey, John H; Carmona-Galindo, Víctor D; Leary, Christopher; Huh, Julie; Valdez, Jennifer

    2013-03-01

    A study was performed in Del Rey Lagoon, City of Los Angeles, to determine if the lagoon was as a source or sink for fecal indicator bacteria (FIB: total coliforms, Escherichia coli, enterococci) and to screen for the presence of other potentially pathogenic bacteria. The lagoon receives tidal flows from the adjacent Ballona Estuary whose water usually is contaminated with FIB originating from the highly urbanized Ballona Creek Watershed. During 16 sampling events from February 2008 through March 2009, replicate water samples (n = 3) were collected 1 h prior to the high tide and 1 h prior to the following low tide. FIB concentrations were measured by the defined substrate method (IDEXX, Westbrook, Me) followed by culturing of bacterial isolates sampled from positive IDEXX Quanti-Tray wells and were identified using the Vitek 2 Compact (bioMérieux, Durham, NC). Mean concentrations of FIB often differed by an order of magnitude from flood to ebb flow conditions. The lagoon tended to act as a sink for total coliforms based on the ratio of mean flood to ebb densities (R (F/E)) >1.0 during 56 % of the sampling events and during ebb flows, as a source for E. coli and enterococci (R (F/E) <1.69 % of events). Approximately 54 species were identified from 277 isolates cultured from the IDEXX Quanti-Trays. Of these, 54 % were species known to include pathogenic strains that can be naturally occurring, introduced in runoff, or originated from other sources. Diversity and cluster analyses indicated a dynamic assemblage that changes in species composition with day-to-day fluctuations as well as tidal action. The concept of monitoring the lagoon and estuary as a sentinel habitat for pathogenic assemblages is discussed.

  3. Effect of Low Energy Waves on the Accumulation and Transport of Fecal Indicator Bacteria in Sand and Pore Water at Freshwater Beaches.

    PubMed

    Wu, Ming Zhi; O'Carroll, Denis M; Vogel, Laura J; Robinson, Clare E

    2017-03-07

    Elevated fecal indicator bacteria (FIB) in beach sand and pore water represent an important nonpoint source of contamination to surface waters. This study examines the physical processes governing the accumulation and distribution of FIB in a beach aquifer. Field data indicate E. coli and enterococci can be transported 1 and 2 m, respectively, below the water table. Data were used to calibrate a numerical model whereby FIB are delivered to a beach aquifer by wave-induced infiltration across the beach face. Simulations indicate FIB rapidly accumulate in a beach aquifer with FIB primarily associated with sand rather than freely residing in the pore water. Simulated transport of E. coli in a beach aquifer is complex and does not correlate with conservative tracer transport. Beaches with higher wave-induced infiltration rate and vertical infiltration velocity (i.e., beaches with higher beach slope and wave height, and lower terrestrial groundwater discharge) had greater E. coli accumulation and E. coli was transported deeper below the beach face. For certain beach conditions, the amount of FIB accumulated in sand over 5-6 days was found to be sufficient to trigger a beach advisory if eroded to surface water.

  4. Microbiological Quality of Ready-to-Eat Vegetables Collected in Mexico City: Occurrence of Aerobic-Mesophilic Bacteria, Fecal Coliforms, and Potentially Pathogenic Nontuberculous Mycobacteria

    PubMed Central

    Cerna-Cortes, Jorge Francisco; Leon-Montes, Nancy; Cortes-Cueto, Ana Laura; Salas-Rangel, Laura P.; Helguera-Repetto, Addy Cecilia; Lopez-Hernandez, Daniel; Rivera-Gutierrez, Sandra; Fernandez-Rendon, Elizabeth; Gonzalez-y-Merchand, Jorge Alberto

    2015-01-01

    The aims of this study were to evaluate the microbiological quality and the occurrence of nontuberculous mycobacteria (NTM) in a variety of salads and sprouts from supermarkets and street vendors in Mexico City. Aerobic-mesophilic bacteria (AMB) were present in 100% of RTE-salads samples; 59% of samples were outside guidelines range (>5.17 log10 CFU per g). Although fecal coliforms (FC) were present in 32% of samples, only 8% of them exceeded the permissible limit (100 MPN/g). Regarding the 100 RTE-sprouts, all samples were also positive for AMB and total coliforms (TC) and 69% for FC. Seven NTM species were recovered from 7 salad samples; they included three M. fortuitum, two M. chelonae, one M. mucogenicum, and one M. sp. Twelve RTE-sprouts samples harbored NTM, which were identified as M. porcinum (five), M. abscessus (two), M. gordonae (two), M. mucogenicum (two), and M. avium complex (one). Most RTE-salads and RTE-sprouts had unsatisfactory microbiological quality and some harbored NTM associated with illness. No correlation between the presence of coliforms and NTM was found. Overall, these results suggest that RTE-salads and RTE-sprouts might function as vehicles for NTM transmission in humans; hence, proper handling and treatment before consumption of such products might be recommendable. PMID:25918721

  5. Microbiological Quality of Ready-to-Eat Vegetables Collected in Mexico City: Occurrence of Aerobic-Mesophilic Bacteria, Fecal Coliforms, and Potentially Pathogenic Nontuberculous Mycobacteria.

    PubMed

    Cerna-Cortes, Jorge Francisco; Leon-Montes, Nancy; Cortes-Cueto, Ana Laura; Salas-Rangel, Laura P; Helguera-Repetto, Addy Cecilia; Lopez-Hernandez, Daniel; Rivera-Gutierrez, Sandra; Fernandez-Rendon, Elizabeth; Gonzalez-y-Merchand, Jorge Alberto

    2015-01-01

    The aims of this study were to evaluate the microbiological quality and the occurrence of nontuberculous mycobacteria (NTM) in a variety of salads and sprouts from supermarkets and street vendors in Mexico City. Aerobic-mesophilic bacteria (AMB) were present in 100% of RTE-salads samples; 59% of samples were outside guidelines range (>5.17 log10 CFU per g). Although fecal coliforms (FC) were present in 32% of samples, only 8% of them exceeded the permissible limit (100 MPN/g). Regarding the 100 RTE-sprouts, all samples were also positive for AMB and total coliforms (TC) and 69% for FC. Seven NTM species were recovered from 7 salad samples; they included three M. fortuitum, two M. chelonae, one M. mucogenicum, and one M. sp. Twelve RTE-sprouts samples harbored NTM, which were identified as M. porcinum (five), M. abscessus (two), M. gordonae (two), M. mucogenicum (two), and M. avium complex (one). Most RTE-salads and RTE-sprouts had unsatisfactory microbiological quality and some harbored NTM associated with illness. No correlation between the presence of coliforms and NTM was found. Overall, these results suggest that RTE-salads and RTE-sprouts might function as vehicles for NTM transmission in humans; hence, proper handling and treatment before consumption of such products might be recommendable.

  6. Characterization of nutrients and fecal indicator bacteria at a concentrated swine feeding operation in Wake County, North Carolina, 2009-2011

    USGS Publications Warehouse

    Harden, Stephen L.; Rogers, Shane W.; Jahne, Michael A.; Shaffer, Carrie E.; Smith, Douglas G.

    2012-01-01

    Study sites were sampled for laboratory analysis of nutrients, total suspended solids (TSS), and (or) fecal indicator bacteria (FIB). Nutrient analyses included measurement of dissolved ammonia, total and dissolved ammonia + organic nitrogen, dissolved nitrate + nitrite, dissolved orthophosphate, and total phosphorus. The FIB analyses included measurement of Escherichia coli and enterococci. Samples of wastewater at the swine facility were collected from a pipe outfall from the swine housing units, two storage lagoons, and the spray fields for analysis of nutrients, TSS, and FIB. Soil samples collected from a spray field were analyzed for FIB. Monitoring locations were established for collecting discharge and water-quality data during storm events at three in-field runoff sites and two sites on the headwater stream (one upstream and one downstream) next to the swine facility. Stormflow samples at the five monitoring locations were collected for four storm events during 2009 to 2010 and analyzed for nutrients, TSS, and FIB. Monthly water samples also were collected during base-flow conditions at all four stream sites for laboratory analysis of nutrients, TSS, and (or) FIB.

  7. Effects of hydrologic, biological, and environmental processes on sources and concentrations of fecal bacteria in the Cuyahoga River, with implications for management of recreational waters in Summit and Cuyahoga Counties, Ohio

    USGS Publications Warehouse

    Myers, Donna N.; Koltun, G.F.; Francy, Donna S.

    1998-01-01

    Discharges of fecal bacteria (fecal coliform bacteria and Escherichia coli ) to the middle main stem of the Cuyahoga River from storm water, combined sewers, and incompletely disinfected wastewater have resulted in frequent exceedances of bacteriological water-quality standards in a 23-mile reach of the river that flows through the Cuyahoga Valley National Recreation Area. Contamination of the middle main stem of the Cuyahoga River by bacteria of fecal origin and subsequent transport to downstream areas where water-contact recreation is an important use of the river are a concern because of the potential public-health risk from the presence of enteric pathogens. Independent field investigations of bacterial decay, dilution, dispersion, transport, and sources, and bacterial contamination of streambed sediments, were completed in 1991-93 during periods of rainfall and runoff. The highest concentration of fecal coliform bacteria observed in the middle main stem during three transport studies exceeded the single-sample fecal coliform standard applicable to primary-contact recreation by a factor of approximately 1,300 and exceeded the Escherichia coli standard by a factor of approximately 8,000. The geometric-mean concentrations of fecal bacteria in the middle main stem were 6.7 to 12.3 times higher than geometric-mean concentrations in the monitored tributaries, and 1.8 to 7.0 times larger than the geometric-mean concentrations discharged from the Akron Water Pollution Control Station. Decay rates of fecal bacteria measured in field studies in 1992 ranged from 0.0018 per hour to 0.0372 per hour for fecal coliform bacteria and from 0.0022 per hour to 0.0407 per hour for Escherichia coli. Most of the decay rates measured in June and August were significantly higher than decay rates measured in April and October. Results of field studies demonstrated that concentrations of fecal coliform bacteria were 1.2 to 58 times higher in streambed sediments than in the overlying

  8. Impact of probiotic administration on the health and fecal microbiota of young calves: a meta-analysis of randomized controlled trials of lactic acid bacteria.

    PubMed

    Signorini, M L; Soto, L P; Zbrun, M V; Sequeira, G J; Rosmini, M R; Frizzo, L S

    2012-08-01

    Before weaning, dairy calves are susceptible to many pathogens which can affect their subsequent performance. The use of lactic acid bacteria (LAB) has been identified as a tool to maintain the intestinal microbial balance and to prevent the establishment of opportunistic pathogenic bacterial populations. However, a consensus has not been reached as to whether probiotics may be effective in reducing the prevalence of gastrointestinal diseases in young calves. The aim of this meta-analysis was to assess the effect of probiotics on diarrhea incidence and the intestinal microbial balance. LAB supplementation has been shown to exert a protective effect and to reduce the incidence of diarrhea (relative risk, RR=0.437, 95% confidence interval (CI) 0.251-0.761). In the subanalysis, this protective effect of the probiotics against diarrhea was observed only in trials that used whole milk (RR=0.154, 95% CI 0.079-0.301) and trials that used multistrain inocula (RR=0.415, 95% CI 0.227-0.759). Probiotics did not improve the fecal characteristics (standardized mean difference, SMD=-0.4904, 95% CI -1.011-0.035) and were unable to change the LAB:coliforms ratio (SMD=0.016, 95% CI -0.701-0.733). Probiotics showed a beneficial impact on the LAB:coliforms ratio in the subanalysis that included trials that used whole milk (SMD=0.780, 95% CI 0.141-1.418) and monostrain inocula (SMD=0.990, 95% CI 0.340-1.641). The probability of significant effects (probiotic positive effect) in a new study was >0.70 for diarrhea and fecal consistency. Whole milk feeding improved the action of the probiotic effect on the incidence of diarrhea and LAB:coliforms ratio. The probability to find significant effects in the diarrhea frequency and LAB:coliforms ratio was higher (P>0.85) if the new studies were conducted using whole milk to feed calves. This paper defines the guidelines to standardize the experimental designs of future trials. LAB can be used as growth promoters in calves instead of antibiotics

  9. Fecal Indicator and Pathogenic Bacteria and Their Antibiotic Resistance in Alluvial Groundwater of an Irrigated Agricultural Region with Dairies.

    PubMed

    Li, Xunde; Atwill, Edward R; Antaki, Elizabeth; Applegate, Olin; Bergamaschi, Brian; Bond, Ronald F; Chase, Jennifer; Ransom, Katherine M; Samuels, William; Watanabe, Naoko; Harter, Thomas

    2015-09-01

    Surveys of microbiological groundwater quality were conducted in a region with intensive animal agriculture in California, USA. The survey included monitoring and domestic wells in eight concentrated animal feeding operations (CAFOs) and 200 small (domestic and community supply district) supply wells across the region. was not detected in groundwater, whereas O157:H7 and were each detected in 2 of 190 CAFO monitoring well samples. Nonpathogenic generic and spp. were detected in 24.2% (46/190) and 97.4% (185/190) groundwater samples from CAFO monitoring wells and in 4.2% (1/24) and 87.5% (21/24) of CAFO domestic wells, respectively. Concentrations of both generic and spp. were significantly associated with well depth, season, and the type of adjacent land use in the CAFO. No pathogenic bacteria were detected in groundwater from 200 small supply wells in the extended survey. However, 4.5 to 10.3% groundwater samples were positive for generic and . Concentrations of generic were not significantly associated with any factors, but concentrations of were significantly associated with proximity to CAFOs, seasons, and concentrations of potassium in water. Among a subset of and isolates from both surveys, the majority of (63.6%) and (86.1%) isolates exhibited resistance to multiple (≥3) antibiotics. Findings confirm significant microbial and antibiotic resistance loading to CAFO groundwater. Results also demonstrate significant attenuative capacity of the unconfined alluvial aquifer system with respect to microbial transport.

  10. Formative Research on Hygiene Behaviors and Geophagy among Infants and Young Children and Implications of Exposure to Fecal Bacteria

    PubMed Central

    Ngure, Francis M.; Humphrey, Jean H.; Mbuya, Mduduzi N. N.; Majo, Florence; Mutasa, Kuda; Govha, Margaret; Mazarura, Exevia; Chasekwa, Bernard; Prendergast, Andrew J.; Curtis, Valerie; Boor, Kathyrn J.; Stoltzfus, Rebecca J.

    2013-01-01

    We conducted direct observation of 23 caregiver–infant pairs for 130 hours and recorded wash-related behaviors to identify pathways of fecal–oral transmission of bacteria among infants. In addition to testing fingers, food, and drinking water of infants, three infants actively ingested 11.3 ± 9.2 (mean ± SD) handfuls of soil and two ingested chicken feces 2 ± 1.4 times in 6 hours. Hand washing with soap was not common and drinking water was contaminated with Escherichia coli in half (12 of 22) of the households. A one-year-old infant ingesting 1 gram of chicken feces in a day and 20 grams of soil from a laundry area of the kitchen yard would consume 4,700,000–23,000,000 and 440–4,240 E. coli, respectively, from these sources. Besides standard wash and nutrition interventions, infants in low-income communities should be protected from exploratory ingestion of chicken feces, soil, and geophagia for optimal child health and growth. PMID:24002485

  11. Prevalence of antibiotic-resistant fecal bacteria in a river impacted by both an antibiotic production plant and urban treated discharges.

    PubMed

    Sidrach-Cardona, Ricardo; Hijosa-Valsero, María; Marti, Elisabet; Balcázar, José Luis; Becares, Eloy

    2014-08-01

    In this study, the abundance and spatial dynamics of antibiotic-resistant fecal bacteria (Escherichia coli, total coliforms and Enterococcus spp.) were determined in water and sediment samples from a river impacted by both antibiotic production plant (APP) and urban wastewater treatment plant (WWTP) discharges. Agar dilution and disk diffusion methods were also used for antimicrobial susceptibility testing. Two antimicrobial agents, cephalexin (25 μg/ml) and amoxicillin (50 μg/ml), were evaluated using the agar dilution method for E. coli, total coliforms (TC) and Enterococcus spp., whereas the degree of sensitivity or resistance of E. coli isolates to penicillin (10 U), ampicillin (10 μg), doxycycline (30 μg), tetracycline (30 μg), erythromycin (15 μg), azithromycin (15 μg) and streptomycin (10 μg) was performed using the disk diffusion method. Real-time PCR assays were used to determine the prevalence of three antibiotic-resistance genes (ARGs). The agar dilution method showed that most E. coli isolates and TC were resistant to amoxicillin, especially after receiving the APP discharges. Antibiotic resistances to amoxicillin and cephalexin were higher after the APP discharge point than after the WWTP effluent. The disk diffusion method revealed that 100% of bacterial isolates were resistant to penicillin and erythromycin. Multidrug-resistant bacteria were detected and showed a higher proportion at the WWTP discharge point than those in the APP. Highly multidrug-resistant bacteria (resistance to more than 4 antibiotics) were also detected, reaching mean values of 41.6% in water samples and 50.1% in sediments. The relative abundance of the blaTEM, blaCTX-M and blaSHV genes was higher in samples from the treatment plants than in those collected upstream from the discharges, especially for water samples collected at the APP discharge point. These results clearly demonstrate that both the APP and the WWTP contribute to the emergence and spread of antibiotic

  12. Quantitative CrAssphage PCR Assays for Human Fecal Pollution Measurement

    EPA Science Inventory

    Environmental waters are monitored for fecal pollution to protect public health and water resources. Traditionally, general fecal indicator bacteria are used; however, they cannot distinguish human fecal waste from pollution from other animals. Recently, a novel bacteriophage, cr...

  13. Effects of feeding wet corn distillers grains with solubles with or without monensin and tylosin on the prevalence and antimicrobial susceptibilities of fecal foodborne pathogenic and commensal bacteria in feedlot cattle.

    PubMed

    Jacob, M E; Fox, J T; Narayanan, S K; Drouillard, J S; Renter, D G; Nagaraja, T G

    2008-05-01

    Distillers grains, a coproduct of ethanol production from cereal grains, are composed principally of the bran, protein, and germ fractions and are commonly supplemented in ruminant diets. The objective of this study was to assess the effect of feeding wet distillers grains with solubles (WDGS) and monensin and tylosin on the prevalence and antimicrobial susceptibilities of fecal foodborne and commensal bacteria in feedlot cattle. Cattle were fed 0 or 25% WDGS in steam-flaked corn-based diets with the addition of no antimicrobials, monensin, or monensin and tylosin. Fecal samples were collected from each animal (n = 370) on d 122 and 136 of the 150-d finishing period and cultured for Escherichia coli O157. Fecal samples were also pooled by pen (n = 54) and cultured for E. coli O157, Salmonella, commensal E. coli, and Enterococcus species. Antimicrobial resistance was assessed by determining antimicrobial susceptibilities of pen bacterial isolates and quantifying antimicrobial resistance genes in fecal samples by real-time PCR. Individual animal prevalence of E. coli O157 in feces collected from cattle fed WDGS was greater (P < 0.001) compared with cattle not fed WDGS on d 122 but not on d 136. There were no treatment effects on the prevalence of E. coli O157 or Salmonella spp. in pooled fecal samples. Antimicrobial susceptibility results showed Enterococcus isolates from cattle fed monensin or monensin and tylosin had greater levels of resistance toward macrolides (P = 0.01). There was no effect of diet or antimicrobials on concentrations of 2 antimicrobial resistance genes, ermB or tetM, in fecal samples. Results from this study indicate that WDGS may have an effect on the prevalence of E. coli O157 and the concentration of selected antimicrobial resistance genes, but does not appear to affect antimicrobial susceptibility patterns in Enterococcus and generic E. coli isolates.

  14. Monitoring urban beaches with qPCR vs. culture measures of fecal indicator bacteria: Implications for public notification.

    PubMed

    Dorevitch, Samuel; Shrestha, Abhilasha; DeFlorio-Barker, Stephanie; Breitenbach, Cathy; Heimler, Ira

    2017-05-12

    The United States Environmental Protection Agency has established methods for testing beach water using the rapid quantitative polymerase chain reaction (qPCR) method, as well as "beach action values" so that the results of such testing can be used to make same-day beach management decisions. Despite its numerous advantages over culture-based monitoring approaches, qPCR monitoring has yet to become widely used in the US or elsewhere. Considering qPCR results obtained on a given day as the best available measure of that day's water quality, we evaluated the frequency of correct vs. incorrect beach management decisions that are driven by culture testing. Beaches in Chicago, USA, were monitored using E. coli culture and enterococci qPCR methods over 894 beach-days in the summers of 2015 and 2016. Agreement in beach management using the two methods, after taking into account agreement due to chance, was summarized using Cohen's kappa statistic. No meaningful agreement (beyond that expected by chance) was observed between beach management actions driven by the two pieces of information available to beach managers on a given day: enterococci qPCR results ofsamples collected that morning and E. coli culture results of samples collected the previous day. The E. coli culture beach action value was exceeded 3.4 times more frequently than the enterococci qPCR beach action value (22.6 vs. 6.6% of beach-days). The largest evaluation of qPCR-based beach monitoring to date provides little scientific rationale for continued E. coli culture testing of beach water in our setting. The observation that the E. coli culture beach action value was exceeded three times as frequently as the enterococci qPCR beach action value suggests that, although the beach action values for bacteria using different measurement methods are thought to provide comparable information about health risk, this does not appear to be the case in all settings.

  15. Fecal impaction

    MedlinePlus

    ... hard and dry. This makes it difficult to pass. Fecal impaction often occurs in people who have ... Small, semi-formed stools Straining when trying to pass stools Other possible symptoms include: Bladder pressure or ...

  16. Fecal Incontinence

    MedlinePlus

    ... Known Home & Community Home › Aging & Health A to Z › Fecal Incontinence Font size A A A Print Share Glossary Basic Facts & Information Causes & Symptoms Diagnosis & Tests Care & Treatment Lifestyle & Management Other Resources Caregiving How ...

  17. Quantitative PCR for Genetic Markers of Human Fecal Pollution

    EPA Science Inventory

    Assessment of health risk and fecal bacteria loads associated with human fecal pollution requires reliable host-specific analytical methods and a rapid quantificationapproach. We report the development of quantitative PCR assays for quantification of two recently described human-...

  18. Quantitative PCR for Genetic Markers of Human Fecal Pollution

    EPA Science Inventory

    Assessment of health risk and fecal bacteria loads associated with human fecal pollution requires reliable host-specific analytical methods and a rapid quantificationapproach. We report the development of quantitative PCR assays for quantification of two recently described human-...

  19. Quantitative PCR for genetic markers of human fecal pollution

    EPA Science Inventory

    Assessment of health risk and fecal bacteria loads associated with human fecal pollution requires reliable host-specific analytical methods and a rapid quantification approach. We report the development of quantitative PCR assays for enumeration of two recently described hum...

  20. Quantitative PCR for genetic markers of human fecal pollution

    EPA Science Inventory

    Assessment of health risk and fecal bacteria loads associated with human fecal pollution requires reliable host-specific analytical methods and a rapid quantification approach. We report the development of quantitative PCR assays for enumeration of two recently described hum...

  1. Biotic interactions and sunlight affect persistence of fecal indicator bacteria and microbial source tracking genetic markers in the upper Mississippi river.

    PubMed

    Korajkic, Asja; McMinn, Brian R; Shanks, Orin C; Sivaganesan, Mano; Fout, G Shay; Ashbolt, Nicholas J

    2014-07-01

    The sanitary quality of recreational waters that may be impacted by sewage is assessed by enumerating fecal indicator bacteria (FIB) (Escherichia coli and enterococci); these organisms are found in the gastrointestinal tracts of humans and many other animals, and hence their presence provides no information about the pollution source. Microbial source tracking (MST) methods can discriminate between different pollution sources, providing critical information to water quality managers, but relatively little is known about factors influencing the decay of FIB and MST genetic markers following release into aquatic environments. An in situ mesocosm was deployed at a temperate recreational beach in the Mississippi River to evaluate the effects of ambient sunlight and biotic interactions (predation, competition, and viral lysis) on the decay of culture-based FIB, as well as molecularly based FIB (Entero1a and GenBac3) and human-associated MST genetic markers (HF183 and HumM2) measured by quantitative real-time PCR (qPCR). In general, culturable FIB decayed the fastest, while molecularly based FIB and human-associated genetic markers decayed more slowly. There was a strong correlation between the decay of molecularly based FIB and that of human-associated genetic markers (r(2), 0.96 to 0.98; P < 0.0001) but not between culturable FIB and any qPCR measurement. Overall, exposure to ambient sunlight may be an important factor in the early-stage decay dynamics but generally was not after continued exposure (i.e., after 120 h), when biotic interactions tended to be the only/major influential determinant of persistence. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  2. Biotic Interactions and Sunlight Affect Persistence of Fecal Indicator Bacteria and Microbial Source Tracking Genetic Markers in the Upper Mississippi River

    PubMed Central

    McMinn, Brian R.; Shanks, Orin C.; Sivaganesan, Mano; Fout, G. Shay; Ashbolt, Nicholas J.

    2014-01-01

    The sanitary quality of recreational waters that may be impacted by sewage is assessed by enumerating fecal indicator bacteria (FIB) (Escherichia coli and enterococci); these organisms are found in the gastrointestinal tracts of humans and many other animals, and hence their presence provides no information about the pollution source. Microbial source tracking (MST) methods can discriminate between different pollution sources, providing critical information to water quality managers, but relatively little is known about factors influencing the decay of FIB and MST genetic markers following release into aquatic environments. An in situ mesocosm was deployed at a temperate recreational beach in the Mississippi River to evaluate the effects of ambient sunlight and biotic interactions (predation, competition, and viral lysis) on the decay of culture-based FIB, as well as molecularly based FIB (Entero1a and GenBac3) and human-associated MST genetic markers (HF183 and HumM2) measured by quantitative real-time PCR (qPCR). In general, culturable FIB decayed the fastest, while molecularly based FIB and human-associated genetic markers decayed more slowly. There was a strong correlation between the decay of molecularly based FIB and that of human-associated genetic markers (r2, 0.96 to 0.98; P < 0.0001) but not between culturable FIB and any qPCR measurement. Overall, exposure to ambient sunlight may be an important factor in the early-stage decay dynamics but generally was not after continued exposure (i.e., after 120 h), when biotic interactions tended to be the only/major influential determinant of persistence. PMID:24747902

  3. Dynamics of fecal indicator bacteria, bacterial pathogen genes, and organic wastewater contaminants in the Little Calumet River: Portage Burns Waterway, Indiana

    USGS Publications Warehouse

    Haack, Sheridan K.; Duris, Joseph W.

    2013-01-01

    Little information exists on the co-occurrence of fecal indicator bacteria (FIB), bacterial pathogens, and organic wastewater-associated chemicals (OWCs) within Great Lakes tributaries. Fifteen watershed sites and one beach site adjacent to the Little Calumet River–Portage Burns Waterway (LCRPBW) on Lake Michigan were tested on four dates for pH, dissolved oxygen, specific conductance, chloride, color, ammonia- and nitrate-nitrogen, soluble phosphorus, sulfate, turbidity, and atrazine; for concentrations of FIB; and for genes indicating the presence of human-pathogenic enterococci (ENT) and of Shiga-toxin producing Escherichia coli (EC) from various animal sources. Nineteen samples were also tested for 60 OWCs. Half of the watershed samples met EC recreational water quality standards; none met ENT standards. Human-wastewater-associated OWC detections were correlated with human-influence indicators such as population/km2, chloride concentrations, and the presence of WWTP effluents, but EC and ENT concentrations were not. Bacterial pathogen genes indicated rural human and several potential animal sources. OWCs of human or ecosystem health concern (musk fragrances AHTN and HHCB, alkylphenols, carbamazepine) and 3 bacterial pathogen genes were detected at the mouth of the LCRPBW, but no such OWCs and only 1 pathogen gene were detected at the beach. The LCRPBW has significant potential to deliver FIB, potential bacterial pathogens, and OWCs of human or ecosystem health concern to the nearshore of Lake Michigan, under conditions enhancing nearshore transport of the river plume. Nearshore mixing of lake and river water, and the lack of relationship between OWCs and FIB or pathogen genes, pose numerous challenges for watershed and nearshore assessment and remediation.

  4. Fecal pollution source tracking in waters intended for human supply based on archaeal and bacterial genetic markers.

    PubMed

    Bianco, Kayo; Barreto, Camila; Oliveira, Samara Sant'Anna; Pinto, Leonardo Henriques; Albano, Rodolpho Mattos; Miranda, Catia Chaia; Clementino, Maysa Mandetta

    2015-12-01

    The determination of fecal pollution sources in aquatic ecosystems is essential to estimate associated health risks. In this study, we evaluate eight microbial source tracking (MST) markers including host-specific Bacteroidales and Methanobrevibacter spp. for discrimination between human, bovine, equine, and swine fecal contamination in waters intended for human supply. Overall, the novel host-specific archaeal and bacterial primers proposed in this study demonstrated high sensitivity and specificity. Markers for the Archaea domain were more prevalent in the fecal and water samples studied. We conclude that the investigations regarding the sources of fecal pollution in public water supplies can contribute to improve the quality of human health. To our knowledge, this is the first analysis using both archaeal and bacterial fecal MST markers on tropical water bodies of Rio de Janeiro city, Brazil.

  5. Enterococcus and Escherichia coli fecal source apportionment with microbial source tracking genetic markers - is it feasible?

    EPA Science Inventory

    Fecal pollution is measured in surface waters using culture-based measurements of enterococci and Escherichia coli bacteria. Source apportionment of these two fecal indicator bacteria is an urgent need for prioritizing remediation efforts and quantifying health risks associated...

  6. Enterococcus and Escherichia coli fecal source apportionment with microbial source tracking genetic markers - is it feasible?

    EPA Science Inventory

    Fecal pollution is measured in surface waters using culture-based measurements of enterococci and Escherichia coli bacteria. Source apportionment of these two fecal indicator bacteria is an urgent need for prioritizing remediation efforts and quantifying health risks associated...

  7. Fecal lactic acid bacteria increased in adolescents randomized to whole-grain but not refined-grain foods, whereas inflammatory cytokine production decreased equally with both interventions.

    PubMed

    Langkamp-Henken, Bobbi; Nieves, Carmelo; Culpepper, Tyler; Radford, Allyson; Girard, Stephanie-Anne; Hughes, Christine; Christman, Mary C; Mai, Volker; Dahl, Wendy J; Boileau, Thomas; Jonnalagadda, Satya S; Thielecke, Frank

    2012-11-01

    The intake of whole-grain (WG) foods by adolescents is reported to be approximately one-third the recommended intake of 48 g/d. This 6-wk randomized interventional study determined the effect of replacing grains within the diet with refined-grain (RG; n = 42) or WG (n = 41) foods/d on gastrointestinal and immune health in adolescents (aged 12.7 ± 0.1 y). A variety of grain-based foods were delivered weekly to participants and their families. Participants were encouraged to eat 3 different kinds of study foods (e.g., bread, cereals, snacks)/d with goals of 0 g/d (RG) and 80 g/d (WG). Stool samples were obtained during the prebaseline and final weeks to measure bifidobacteria and lactic acid bacteria (LAB) using qPCR. Stool frequency was recorded daily. Blood was drawn at baseline and at final visits for immune markers. Across groups, total-grain intake increased by one serving. The intake of WG was similar at baseline (18 ± 3 g) between groups but increased to 60 ± 5 g in the WG group and decreased to 4 ± 1 g in the RG group. Fecal bifidobacteria increased from baseline with both interventions, but LAB increased (P < 0.05) from baseline [2.4 ± 0.2 log(10) genome equivalents (eq)] to wk 6 (3.0 ± 0.2 log(10) genome eq) in the WG group but not in the RG group (baseline: 2.9 ± 0.2 log(10) genome eq; wk 6: 3.0 ± 0.1 log(10) genome eq). There was no difference in stool frequency, serum antioxidant potential, or in vitro LPS-stimulated mononuclear cell production of inflammatory cytokines between groups. However, across both groups the number of daily stools tended to increase (P = 0.08) by 0.0034 stools/g WG or by 0.2 stools with 60 g WG, mean antioxidant potential increased by 58%, and mean production of TNF-α, IL-1β, and IL-6 decreased by 24, 22, and 42%, respectively, between baseline and wk 6. Overall, incorporating either WG or RG foods increased serum antioxidant concentrations and decreased inflammatory cytokine production; however, WG study foods had

  8. Vegetable fiber fermentation by human fecal bacteria: cell wall polysaccharide disappearance and short-chain fatty acid production during in vitro fermentation and water-holding capacity of unfermented residues.

    PubMed

    Bourquin, L D; Titgemeyer, E C; Fahey, G C

    1993-05-01

    Dietary fiber from eight vegetables (broccoli, carrot, cauliflower, celery, cucumber, lettuce, onion and radish) was analyzed for chemical composition and potential in vitro fermentation by human fecal bacteria. Total dietary fiber concentration of substrates ranged from 34.9 (broccoli) to 5.8 (cucumber) g/kg edible matter. Substrate fiber fractions were composed primarily of pectic substances and cellulose with smaller concentrations of hemicelluloses and lignin. Total dietary fiber residues isolated from substrates were fermented in vitro for 24 h with fecal bacteria obtained from each of three human volunteers. Substrate dry matter disappearance during fermentation was highest for carrot (63.7%) and lowest for cucumber (49.4%). Averaged across all substrates, disappearances of arabinose, galactose, glucose, mannose, xylose and uronic acids during fermentation were 96, 90, 54, 68, 51 and 97%, respectively. Short-chain fatty acid (SCFA) production during substrate fermentation averaged 10.5 mmol SCFA/g dry matter fermented. Averaged across all substrates, production of the major SCFA, acetate, propionate and butyrate, occurred in the molar ratio 76:14:10. Potential water-holding capacity of substrates was not influenced by fiber source and averaged 2.04 g H2O/g original substrate dry matter. Extent of substrate fermentation, SCFA production and substrate potential water-holding capacity were significantly different among inoculum donors, indicating that considerable inter-individual variation exists in the potential in vivo fermentation of vegetable fiber.

  9. Use of radon-222 to evaluate the influence of groundwater discharge on fecal indicator bacteria concentrations in the near-shore ocean, Malibu, California

    NASA Astrophysics Data System (ADS)

    Izbicki, J. A.; Burton, C.; Swarzenski, P. W.

    2011-12-01

    To protect beach-goers from waterborne disease, California requires water-quality monitoring for fecal indicator bacteria (FIB) at beaches having more than 50,000 visits annually. The source(s) of FIB in ocean beaches in excess of marine recreational water standards is often not known, or may be incorrectly identified. Onsite wastewater treatment systems (OWTS) used to treat residential and commercial sewage have been implicated by regulatory agencies as a possible source of FIB to recreational ocean beaches, near Malibu, California. For this to occur, treated wastewater must first move through groundwater prior to discharge at the ocean. Groundwater discharge to the ocean near Malibu Lagoon (the estuary of Malibu Creek) is complicated by seasonally changing water levels in the lagoon. The lagoon is isolated from the ocean by a sand berm that develops across the mouth of the lagoon during the dry season. Higher water levels in the lagoon during the dry season, and lower water-levels during the wet season, cause seasonal changes in the direction of groundwater flow and the magnitude of discharge from the adjacent small (3,400 hectare), alluvial aquifer. Radon-222, an indicator of groundwater discharge, was measured in Malibu Lagoon, in the near-shore ocean adjacent to the lagoon, and in the near-shore ocean adjacent to unsewered residential development to determine the timing and magnitude of groundwater discharge. During the dry season, when the berm of the lagoon was closed and the lagoon was isolated from the ocean, radon-222 concentrations in the near-shore ocean during low tide increased as water discharged from the lagoon through the berm. Enterococcus concentrations in the near-shore ocean increased to almost 600 Most Probable Number (MPN) per 100 milliliter at this time. Radon-222 concentrations also increased at low tide as groundwater discharged to the ocean from the adjacent alluvial aquifer underlying the unsewered residential development, but there was

  10. MOLECULAR EVALUATION OF CHANGES IN PLANKTONIC BACTERIAL POPULATION RESULTING FROM EQUINE FECAL CONTAMINATION IN A SUB-WATERSHED

    EPA Science Inventory

    Contamination of watersheds by fecal bacteria is a frequent cause for surface waters to be placed on the national impaired waters list. However, since the presence of fecal bacteria does not always indicate human fecal input, it is necessary to distinguish between fecal sources. ...

  11. MOLECULAR EVALUATION OF CHANGES IN PLANKTONIC BACTERIAL POPULATION RESULTING FROM EQUINE FECAL CONTAMINATION IN A SUB-WATERSHED

    EPA Science Inventory

    Contamination of watersheds by fecal bacteria is a frequent cause for surface waters to be placed on the national impaired waters list. However, since the presence of fecal bacteria does not always indicate human fecal input, it is necessary to distinguish between fecal sources. ...

  12. Occurrence and trends in the concentrations of fecal-indicator bacteria and the relation to field water-quality parameters in the Allegheny, Monongahela, and Ohio Rivers and selected tributaries, Allegheny County, Pennsylvania, 2001–09

    USGS Publications Warehouse

    Fulton, John W.; Koerkle, Edward H.; McCoy, Jamie L.; Zarr, Linda F.

    2016-01-21

    A total of 1,742 water samples were collected at 52 main-stem and tributary sites. Quantifiable concentrations of Escherichia coli (E. coli) were reported in 1,667 samples, or 97.0 percent of 1,719 samples; concentrations in 853 samples (49.6 percent) exceeded the U.S. Environmental Protection Agency (EPA) recreational water-quality criterion of 235 colonies per 100 milliliters (col/100 mL). Quantifiable concentrations of fecal coliform (FC) bacteria were reported in 1,693 samples, or 98.8 percent of 1,713 samples; concentrations in 780 samples (45.5 percent) exceeded the Commonwealth of Pennsylvania water contact criterion of 400 col/100 mL. Quantifiable concentrations of enterococci bacteria were reported in 912 samples, or 87.5 percent of 1,042 samples; concentrations in 483 samples (46.4 percent) exceeded the EPA recreational water-quality criterion of 61 col/100 mL. The median percentage of samples in which bacteria concentrations exceeded recreational water-quality standards across all sites with five or more samples was 48 for E. coli, 43 for FC, and 75 for enterococci. E. coli, FC, and enterococci concentrations at main-stem sites had significant positive correlations with streamflow under all weather conditions, with rho values ranging from 0.203 to 0.598. Seasonal Kendall and logistic regression were evaluated to determine whether statistically significant trends were present during the period 2001–09. In general, Seasonal Kendall tests for trends in E. coli and FC bacteria were inconclusive. Results of logistic regression showed no significant trends in dry-weather exceedance of the standards; however, significant decreases in the likelihood that wet-weather E. coli and FC bacteria concentrations will exceed EPA recreational standards were found at the USGS streamgaging station Allegheny River at 9th Street Bridge. Nonparametric correlation analysis, including Spearman’s rho and the paired Prentice-Wilcoxon test, was used to screen for associations

  13. Effects of receiving-water quality and wastewater treatment on injury, survival, and regrowth of fecal-indicator bacteria and implications for assessment of recreational water quality

    USGS Publications Warehouse

    Francy, D.S.; Hart, T.L.; Virosteck, C.M.

    1996-01-01

    Bacterial injury, survival, and regrowth were investigated by use of replicate flow-through incubation chambers placed in the Cuyahoga River or Lake Erie in the greater Cleveland metropolitan area during seven 4-day field studies. The chambers contained wastewater or combined-sewer-overflow (CSO) effluents treated three ways-unchlorinated, chlorinated, and dechlorinated. At timestep intervals, the chamber contents were analyzed for concentrations of injured and healthy fecal coliforms by use of standard selective and enhanced-recovery membrane-filtration methods. Mean percent injuries and survivals were calculated from the fecal-coliform concentration data for each field study. The results of analysis of variance (ANOVA) indicated that treatment affected mean percent injury and survival, whereas site did not. In the warm-weather Lake Erie field study, but not in the warm-weather Cuyahoga River studies, the results of ANOVA indicated that dechlorination enhanced the repair of injuries and regrowth of chlorine-injured fecal coliforms on culture media over chlorination alone. The results of ANOVA on the percent injury from CSO effluent field studies indicated that dechlorination reduced the ability of organisms to recover and regrow on culture media over chlorination alone. However, because of atypical patterns of concentration increases and decreases in some CSO effluent samples, more work needs to be done before the effect of dechlorination and chlorination on reducing fecal-coliform concentrations in CSO effluents can be confirmed. The results of ANOVA on percent survivals found statistically significant differences among the three treatment methods for all but one study. Dechlorination was found to be less effective than chlorination alone in reducing the survival of fecal coliforms in wastewater effluent, but not in CSO effluent. If the concentration of fecal coliforms determined by use of the enhanced-recovery method can be predicted accurately from the

  14. The effect of dietary phosphorus and calcium level, phytase supplementation, and ileal infusion of pectin on the chemical composition and carbohydrase activity of fecal bacteria and the level of microbial metabolites in the gastrointestinal tract of pigs.

    PubMed

    Metzler, B U; Mosenthin, R; Baumgärtel, T; Rodehutscord, M

    2008-07-01

    Two experiments with growing pigs were conducted to determine the effects of dietary P and Ca level, phytase supplementation, and ileal pectin infusion on ileal and fecal P and Ca balance, chemical composition of fecal mixed bacterial mass (MBM), and bacterial metabolic activity. Pigs (initial BW = 30 kg) were fitted with simple T-cannulas at the distal ileum. They were fed a low-P corn-soybean meal control diet (3 g of P/kg) or the control diet supplemented with monocalcium phosphate (MCP; 7 g of P/kg; Exp. 1) or 1,000 FTU phytase/kg (Exp. 2). The daily infusion treatments consisted of 60 g of pectin dissolved in 1.8 L of demineralized water or 1.8 L of demineralized water as the control infusion, infused via the ileal cannula. In each experiment, 8 barrows were assigned to 4 dietary treatments according to a double, incomplete 4 x 2 Latin square. The dietary treatments in Exp. 1 were the control (Con-) diet with water infusion; the control (Con+) diet with pectin infusion; the MCP diet with water infusion; and the MCP diet with pectin infusion. In Exp. 2, the pigs received the same Con- and Con+ treatments as in Exp. 1 and, in addition, the phytase-supplemented diet in combination with water or pectin infusion. After a 15-d adaptation period, feces were collected for 5 d followed by ileal digesta collection for 24 h. In Exp. 1, supplemental MCP increased (P fecal P and Ca recovery as well as P and Ca content of the MBM. Pectin infusion increased the N content of the MBM (P = 0.054) and polygalacturonase activity (P = 0.032) in feces. In addition, pectin decreased (P = 0.049) ileal and tended (P < 0. 079) to increase fecal VFA concentrations. In Exp. 2, phytase decreased ileal and fecal P recovery (P < 0.001) and the P content of the MBM (P = 0.045), whereas the N content of the MBM (P = 0.094) and fecal cellulase activity (P = 0.089) tended to decrease. Similarly, pectin infusion decreased (P = 0.036) fecal cellulase activity but increased (P

  15. Composition of fecal microbiota of laboratory mice derived from Japanese commercial breeders using 16S rRNA gene clone libraries

    PubMed Central

    NOZU, Ryoko; UENO, Masami; HAYASHIMOTO, Nobuhito

    2016-01-01

    The fecal microbiota of six mice derived from three Japanese commercial breeders was analyzed by using 16S rRNA gene clone libraries to construct a database for analyzing the gut microbiota of laboratory mice. The 566 clones were obtained from the clone libraries generated from the fecal DNA samples derived from BALB/c, C57BL/6N, DBA/2 and ICR mice. Among these 566 clones, there were 446 unique 16S rRNA gene sequences. When grouped at the 98% similarity level, the 446 unique sequences consisted of 103 Clostridiales, 43 Bacteroidales, 5 Lactobacillus and 3 Erysipelotricaceae, as well as sequences from 11 other phyla. PMID:26902692

  16. Interaction of fecal coliforms with soil aggregates

    USDA-ARS?s Scientific Manuscript database

    Land-applied manures may contain various contaminants that cause water pollution and concomitant health problems. Some of these pollutants are bacteria, and fecal coliforms (FC) have been widely used as an indicator of bacterial contamination. Experiments on bacteria attachment to soil are tradition...

  17. Human-, Ovine-, and Bovine-Specific Viral Source Tracking Tools to Discriminate Between the Major Fecal Sources in Agricultural Waters.

    PubMed

    Rusiñol, Marta; Moriarty, Elaine; Lin, Susan; Bofill-Mas, Sílvia; Gilpin, Brent

    2016-03-01

    This study evaluated the sources of fecal contamination in different river catchments, using a combination of microbial source tracking tools, for human, ruminant, ovine and bovine livestock, in order to define appropriate water management strategies. Every source of waterway pollution was evaluated in river water samples from one urban river catchment and two important farming regions in New Zealand. Fecal pollution was initially measured by testing Escherichia coli and evaluating the presence of human- and ruminant-associated DNA markers of Bacteroidales (BiAdo, BacHum-UCD, BacH, and BacR) and human and ruminant fecal sterols/stanols ratios. Then specific fecal pollution sources were assessed with previously reported quantitative PCR assays targeting human-, bovine-, and ovine-specific viruses: human adenoviruses (HAdV), human JC polyomaviruses, bovine polyomaviruses (BPyV), and ovine polyomaviruses (OPyV). High level of ruminant fecal contamination was detected all over the farming areas, whereas no ruminant sources were identified in the urban river sampling sites. BacR was the most frequently observed ruminant marker and OPyV and BPyV allowed the identification of ovine and bovine fecal sources. The human fecal viral marker (HAdV) was the most frequently observed human marker, highly abundant in the urban sites, and also present in farming areas. This is the first study using simultaneously the ovine and the bovine viral markers to identify and quantify both bovine and ovine fecal pollution.

  18. Correlation between the human fecal microbiota and depression.

    PubMed

    Naseribafrouei, A; Hestad, K; Avershina, E; Sekelja, M; Linløkken, A; Wilson, R; Rudi, K

    2014-08-01

    Depression is a chronic syndrome with a pathogenesis linked to various genetic, biological, and environmental factors. Several links between gut microbiota and depression have been established in animal models. In humans, however, few correlations have yet been demonstrated. The aim of our work was therefore to identify potential correlations between human fecal microbiota (as a proxy for gut microbiota) and depression. We analyzed fecal samples from 55 people, 37 patients, and 18 non-depressed controls. Our analyses were based on data generated by Illumina deep sequencing of 16S rRNA gene amplicons. We found several correlations between depression and fecal microbiota. The correlations, however, showed opposite directions even for closely related Operational Taxonomic Units (OTU's), but were still associated with certain higher order phylogroups. The order Bacteroidales showed an overrepresentation (p = 0.05), while the family Lachnospiraceae showed an underrepresentation (p = 0.02) of OTU's associated with depression. At low taxonomic levels, there was one clade consisting of five OTU's within the genus Oscillibacter, and one clade within Alistipes (consisting of four OTU's) that showed a significant association with depression (p = 0.03 and 0.01, respectively). The Oscillibacter type strain has valeric acid as its main metabolic end product, a homolog of neurotransmitter GABA, while Alistipes has previously been shown to be associated with induced stress in mice. In conclusion, the taxonomic correlations detected here may therefore correspond to mechanistic models. © 2014 John Wiley & Sons Ltd.

  19. Differential decay of Enterococci and Escherichia coli originating from two fecal pollution sources

    EPA Science Inventory

    Using in situ subtropical aquatic mesocosms, fecal source (cattle manure versus sewage) was shown to be the most important contributor to differential loss in viability of fecal indicator bacteria (FIB), specifically enterococci in freshwater and Escherichia coli in marine habita...

  20. Differential decay of Enterococci and Escherichia coli originating from two fecal pollution sources

    EPA Science Inventory

    Using in situ subtropical aquatic mesocosms, fecal source (cattle manure versus sewage) was shown to be the most important contributor to differential loss in viability of fecal indicator bacteria (FIB), specifically enterococci in freshwater and Escherichia coli in marine habita...

  1. Hands, water, and health: fecal contamination in Tanzanian communities with improved, non-networked water supplies.

    PubMed

    Pickering, Amy J; Davis, Jennifer; Walters, Sarah P; Horak, Helena M; Keymer, Daniel P; Mushi, Douglas; Strickfaden, Rachelle; Chynoweth, Joshua S; Liu, Jessie; Blum, Annalise; Rogers, Kirsten; Boehm, Alexandria B

    2010-05-01

    Almost half of the world's population relies on non-networked water supply services, which necessitates in-home water storage. It has been suggested that dirty hands play a role in microbial contamination of drinking water during collection, transport, and storage. However, little work has been done to evaluate quantitatively the association between hand contamination and stored water quality within households. This study measured levels of E. coli, fecal streptococci, and occurrence of the general Bacteroidales fecal DNA marker in source water, in stored water, and on hands in 334 households among communities in Dar es Salaam, Tanzania, where residents use non-networked water sources. Levels of fecal contamination on hands of mothers and children were positively correlated to fecal contamination in stored drinking water within households. Household characteristics associated with hand contamination included mother's educational attainment, use of an improved toilet, an infant in the household, and dissatisfaction with the quantity of water available for hygiene. In addition, fecal contamination on hands was associated with the prevalence of gastrointestinal and respiratory symptoms within a household. The results suggest that reducing fecal contamination on hands should be investigated as a strategy for improving stored drinking water quality and health among households using non-networked water supplies.

  2. Molecular Detection of Campylobacter spp. and Fecal Indicator Bacteria during the Northern Migration of Sandhill Cranes (Grus canadensis) at the Central Platte River

    PubMed Central

    Ryu, Hodon; Vogel, Jason; Santo Domingo, Jorge; Ashbolt, Nicholas J.

    2013-01-01

    The risk to human health of the annual sandhill crane (Grus canadensis) migration through Nebraska, which is thought to be a major source of fecal pollution of the central Platte River, is unknown. To better understand potential risks, the presence of Campylobacter species and three fecal indicator bacterial groups (Enterococcus spp., Escherichia coli, and Bacteroidetes) was assayed by PCR from crane excreta and water samples collected during their stopover at the Platte River, Nebraska, in 2010. Genus-specific PCR assays and sequence analyses identified Campylobacter jejuni as the predominant Campylobacter species in sandhill crane excreta. Campylobacter spp. were detected in 48% of crane excreta, 24% of water samples, and 11% of sediment samples. The estimated densities of Enterococcus spp. were highest in excreta samples (mean, 4.6 × 108 cell equivalents [CE]/g), while water samples contained higher levels of Bacteroidetes (mean, 5.1 × 105 CE/100 ml). Enterococcus spp., E. coli, and Campylobacter spp. were significantly increased in river water and sediments during the crane migration period, with Enterococcus sp. densities (∼3.3 × 105 CE/g) 2 to 4 orders of magnitude higher than those of Bacteroidetes (4.9 × 103 CE/g), E. coli (2.2 × 103 CE/g), and Campylobacter spp. (37 CE/g). Sequencing data for the 16S rRNA gene and Campylobacter species-specific PCR assays indicated that C. jejuni was the major Campylobacter species present in water, sediments, and crane excreta. Overall, migration appeared to result in a significant, but temporary, change in water quality in spring, when there may be a C. jejuni health hazard associated with water and crops visited by the migrating birds. PMID:23584775

  3. [Fecal microbiota transplantation].

    PubMed

    García-García-de-Paredes, Ana; Rodríguez-de-Santiago, Enrique; Aguilera-Castro, Lara; Ferre-Aracil, Carlos; López-Sanromán, Antonio

    2015-03-01

    Bacteria can no longer be seen as an enemy. Nowadays, there is enough evidence to place the microbiota as a key element in human homeostasis. Despite initial skepticism, fecal microbiota transplantation (FMT) is a real therapeutic alternative for patients with recurrent Clostridium difficile infection. Moreover, this procedure has shown promising results in ulcerative colitis and other non-gastrointestinal disorders. There is still a lack of knowledge and clinical trials with long- term follow-up. Therefore, the available data should be interpreted with caution. In this document we provide a detailed review of the literature on the intestinal microbiota and FMT. Copyright © 2014 Elsevier España, S.L.U. and AEEH y AEG. All rights reserved.

  4. Effects of tillage and poultry manure application rates on Salmonella and fecal indicator bacteria concentrations in tiles draining Des Moines Lobe soils

    USDA-ARS?s Scientific Manuscript database

    Application of poultry manure (PM) to cropland as fertilizer is a common practice in artificially drained regions of the Upper Midwest. To assess the potential for PM to contribute pathogenic bacteria to downstream waters, information is needed on the impacts of manure management and tillage practi...

  5. Removal of Fecal Indicators, Pathogenic Bacteria, Adenovirus, Cryptosporidium and Giardia (oo)cysts in Waste Stabilization Ponds in Northern and Eastern Australia

    PubMed Central

    Sheludchenko, Maxim; Padovan, Anna; Katouli, Mohammad; Stratton, Helen

    2016-01-01

    Maturation ponds are used in rural and regional areas in Australia to remove the microbial loads of sewage wastewater, however, they have not been studied intensively until present. Using a combination of culture-based methods and quantitative real-time PCR, we assessed microbial removal rates in maturation ponds at four waste stabilization ponds (WSP) with (n = 1) and without (n = 3) baffles in rural and remote communities in Australia. Concentrations of total coliforms, E. coli, enterococci, Campylobacter spp., Salmonella spp., F+ RNA coliphage, adenovirus, Cryptosporidium spp. and Giardia (oo) cysts in maturation ponds were measured at the inlet and outlet. Only the baffled pond demonstrated a significant removal of most of the pathogens tested and therefore was subjected to further study by analyzing E. coli and enterococci concentrations at six points along the baffles over five sampling rounds. Using culture-based methods, we found a decrease in the number of E. coli and enterococci from the initial values of 100,000 CFU per 100 mL in the inlet samples to approximately 1000 CFU per 100 mL in the outlet samples for both bacterial groups. Giardia cysts removal was relatively higher than fecal indicators reduction possibly due to sedimentation. PMID:26729150

  6. Removal of Fecal Indicators, Pathogenic Bacteria, Adenovirus, Cryptosporidium and Giardia (oo)cysts in Waste Stabilization Ponds in Northern and Eastern Australia.

    PubMed

    Sheludchenko, Maxim; Padovan, Anna; Katouli, Mohammad; Stratton, Helen

    2016-01-02

    Maturation ponds are used in rural and regional areas in Australia to remove the microbial loads of sewage wastewater, however, they have not been studied intensively until present. Using a combination of culture-based methods and quantitative real-time PCR, we assessed microbial removal rates in maturation ponds at four waste stabilization ponds (WSP) with (n = 1) and without (n = 3) baffles in rural and remote communities in Australia. Concentrations of total coliforms, E. coli, enterococci, Campylobacter spp., Salmonella spp., F+ RNA coliphage, adenovirus, Cryptosporidium spp. and Giardia (oo) cysts in maturation ponds were measured at the inlet and outlet. Only the baffled pond demonstrated a significant removal of most of the pathogens tested and therefore was subjected to further study by analyzing E. coli and enterococci concentrations at six points along the baffles over five sampling rounds. Using culture-based methods, we found a decrease in the number of E. coli and enterococci from the initial values of 100,000 CFU per 100 mL in the inlet samples to approximately 1000 CFU per 100 mL in the outlet samples for both bacterial groups. Giardia cysts removal was relatively higher than fecal indicators reduction possibly due to sedimentation.

  7. Environmental factors influencing the distribution of total and fecal coliform bacteria in six water storage reservoirs in the Pearl River Delta Region, China.

    PubMed

    Hong, Huachang; Qiu, Jianwen; Liang, Yan

    2010-01-01

    The Pearl River Delta (PRD) is one of the most developed and densely populated regions in China. Quantifying the amount of pathogens in the source of drinking water is important for improving water quality. We collected water samples from six major water storage reservoirs in the PRD region in both wet and dry seasons in 2006. Results showed that external environmental factors, such as precipitation, location, as well as the internal environmental factors, i.e., physicochemical properties of the water, were closely related with the distribution of coliforms. Seasonally, the coliform bacterial concentrations in wet season were one to two orders of magnitude greater than those in dry season. Spatially, coliform bacterial levels in reservoirs near urban and industrial areas were significantly higher (p < 0.05) than those in remote areas. Correlation analyses showed that the levels of coliforms had close relationships with pH, temperature, suspended solid, organic and inorganic nutrients in water. Principal components analysis further demonstrated that total coliforms in the reservoirs were closely related with water physicochemical properties, while fecal coliforms were more associated with external input brought in by seasonal runoff.

  8. Influence of source and concentrations of dietary fiber on in vivo nitrogen excretion pathways in pigs as reflected by in vitro fermentation and nitrogen incorporation by fecal bacteria.

    PubMed

    Bindelle, J; Buldgen, A; Delacollette, M; Wavreille, J; Agneessens, R; Destain, J P; Leterme, P

    2009-02-01

    The inclusion of dietary fiber (DF) in diets has been suggested as a way to reduce NH(3) emission in pig barns because it contributes to a shift in N excretion from urine to feces owing to enhanced bacterial growth in the intestines. This study compared an in vitro method to measure bacterial protein synthesis during fermentation with an in vivo N excretion shift induced by diets differing in DF concentrations and solubility. The first experiment measured the effect of graded concentrations of sugar beet pulp (SBP; 0, 10, 20, and 30%) in corn- and soybean meal-based diets on in vivo N excretion partitioning between the urine and feces. A second experiment investigated the replacement of SBP, rich in soluble DF, with oat hulls (OH), rich in insoluble DF (20:0, 10.5:10.5, and 0:22%, respectively). In parallel, the fermentation characteristics of the dietary carbohydrates not digested in the small intestine were evaluated in an in vitro gas test, based on their incubation with colonic microbiota, using a mineral buffer solution enriched with (15)N. The N originating from the buffer solution incorporated into the bacterial proteins (BNI) was measured when half the final gas volume was produced (8.5 to 14.5 h of fermentation) and after 72 h of fermentation. Short-chain fatty acids were determined in the liquid phase. In the first experiment, the inclusion of SBP linearly decreased urinary N excretion from 0.285 to 0.215 g of N excreted in the urine per gram of N ingested and decreased the urinary-N:fecal-N excretion ratio from 2.171 to 1.177 (P < 0.01). In the second experiment, substituting SBP with OH linearly increased the urinary-N:fecal-N excretion ratio (P = 0.009). Unlike short-chain fatty acid production, BNI was greater at half-time to asymptotic gas production than at 72 h of fermentation. Sugar beet pulp enhanced BNI linearly (P < 0.001), 2.01, 2.06, and 2.35 mg g(-1) of diet with 10, 20, and 30% SBP, respectively, as compared with 1.51 mg for the control

  9. Dynamics in Sand of Fecal Indicator Bacteria (FIB) and Salmonella From Contaminated Water, Runoff, and Sewage in an Urbanized Southern California Shoreline

    NASA Astrophysics Data System (ADS)

    Mika, K.; Lee, C.; Lin, C.; Imamura, G.; Chang, C.; Jay, J.

    2007-12-01

    In urbanized coastal watersheds, FIB can come from a variety of sources, including contaminated freshwater sources such as storm drains or creeks, sewage spills, and overlying waterbodies. To investigate the impact of these sources on bacterial levels in coastal sediments and resultant impacts on water bodies, we studied these three source types in Southern California. First, we sampled at 8am and noon at three locations throughout the summer at an enclosed beach on Avalon Bay, Catalina Island, in collaboration with Southern California Coastal Water Research Project. Using membrane filtration and IDEXX substrate technology, we measured FIB levels in sediment and water and observed a positive correlation between these two factors. Second, we studied bacterial persistence in sediment using human sewage as source. To study bacterial survival in the natural environment, we tested solar disinfection with raking as a disinfection procedure after a large sewage spill in the Los Angeles area. First order decay constants for the E. coli ranged between -0.23 and -1.02 in test plots. Further bench scale studies were conducted to determine the effect of various factors on inactivation kinetics. Decay constants measured in controlled experiments in both February and June on the rooftop ranged from -0.73 to -1.54 even though both temperature and intensity of sunlight varied greatly in these months. Interestingly, microcosms subjected to constant moisture had similar decay constants for enterococci, while E. coli in moistened sand showed very low decay rates. Finally, we further investigated effects of sunlight by assessing the extent of surface sand contamination near two contaminated freshwater sources, one under constant shading, and looked for evidence of human fecal matter.

  10. Water-quality assessment of the upper Illinois River Basin in Illinois, Indiana, and Wisconsin; nutrients, dissolved oxygen, and fecal-indicator bacteria in surface water, April 1987 through August 1990

    USGS Publications Warehouse

    Terrio, P.J.

    1995-01-01

    Data describing the presence, spatial distribution, and temporal variability of nutrients, dissolved oxygen, and fecal-indicator bacteria in surface water were collected from streams in the upper Illinois River Basin from 1987-90 as part of the U.S. Geological Survey's National Water-Quality Assessment (NAWQA) program. The largest concen- trations and loads of total nitrogen and total phosphorus were observed in streams in the urban areas of the basin. Mean annual loads of total nitrogen and total phosphorus leaving the upper Illinois River Basin accounted for 30 and 4 percent, respectively, of the input of these nutrients to the basin. Upward trends in total nitrogen concen- trations from 1978-90 were observed at three surface-water sampling stations, and downward trends in total phosphorus concentrations were observed at two stations. Median dissolved oxygen concentrations ranged from 3.4 to 12.2 milligrams per liter at eight long-term monitoring stations in the basin. During low-flow conditions, dissolved oxygen concentrations at 59 percent of the sites in the agricultural Kankakee River Basin and 49 percent of the sites in the urban Des Plaines River Basin were less than the Illinois water-quality standard of 5.0 milligrams per liter. Upward trends in dissolved oxygen concentrations were indicated at the two most downstream stations in the upper Illinois River Basin. Fecal-coliform densities at the fixed stations ranged from 1 to 45,000 colonies per 100 milliliters; stream-water samples from the Des Plaines River Basin typically had densities one or two orders of magnitude larger than samples from the rest of the Upper Illinois River Basin. Between 30 and 100 percent of the samples collected at surface-water sampling stations in the Des Plaines River Basin had densities of E.Coli greater than the Federal criteria for infrequently used full-body- contact water. Significant downward trends in bacteria densities were observed at three of the surface

  11. Blautia and Prevotella sequences distinguish human and animal fecal pollution in Brazil surface waters.

    PubMed

    Koskey, Amber M; Fisher, Jenny C; Eren, A Murat; Ponce-Terashima, Rafael; Reis, Mitermayer G; Blanton, Ronald E; McLellan, Sandra L

    2014-12-01

    Untreated sewage discharges and limited agricultural manure management practices contribute to fecal pollution in rural Brazilian waterways. Most microbial source tracking studies have focused on Bacteroidales, and few have tested host-specific indicators in underdeveloped regions. Sequencing of sewage and human and animal feces with Illumina HiSeq revealed Prevotellaceae as the most abundant family in humans, with Lachnospiraceae and Ruminococcaceae also comprising a large proportion of the microbiome. These same families were also dominant in animals. Bacteroides, the genus containing the most commonly utilized human-specific marker in the United States was present in very low abundance. We used oligotyping to identify Prevotella and Blautia sequences that can distinguish human fecal contamination. Thirty-five of 61 Blautia oligotypes and 13 of 108 Prevotella oligotypes in humans were host-specific or highly abundant (i.e. host-preferred) compared to pig, dog, horse and cow sources. Certain human Prevotella and Blautia oligotypes increased more than an order of magnitude along a polluted river transect in rural Brazil, but traditional fecal indicator levels followed a steady or even decreasing trend. While both Prevotella and Blautia oligotypes distinguished human and animal fecal pollution in Brazil surface waters, Blautia appears to contain more discriminatory and globally applicable markers for tracking sources of fecal pollution.

  12. Occurrence and distribution of fecal indicator bacteria, and physical and chemical indicators of water quality in streams receiving discharge from Dallas/Fort Worth International Airport and vicinity, North-Central Texas, 2008

    USGS Publications Warehouse

    Harwell, Glenn R.; Mobley, Craig A.

    2009-01-01

    This report, done by the U.S. Geological Survey in cooperation with Dallas/Fort Worth International (DFW) Airport in 2008, describes the occurrence and distribution of fecal indicator bacteria (fecal coliform and Escherichia [E.] coli), and the physical and chemical indicators of water quality (relative to Texas Surface Water Quality Standards), in streams receiving discharge from DFW Airport and vicinity. At sampling sites in the lower West Fork Trinity River watershed during low-flow conditions, geometric mean E. coli counts for five of the eight West Fork Trinity River watershed sampling sites exceeded the Texas Commission on Environmental Quality E. coli criterion, thus not fully supporting contact recreation. Two of the five sites with geometric means that exceeded the contact recreation criterion are airport discharge sites, which here means that the major fraction of discharge at those sites is from DFW Airport. At sampling sites in the Elm Fork Trinity River watershed during low-flow conditions, geometric mean E. coli counts exceeded the geometric mean contact recreation criterion for seven (four airport, three non-airport) of 13 sampling sites. Under low-flow conditions in the lower West Fork Trinity River watershed, E. coli counts for airport discharge sites were significantly different from (lower than) E. coli counts for non-airport sites. Under low-flow conditions in the Elm Fork Trinity River watershed, there was no significant difference between E. coli counts for airport sites and non-airport sites. During stormflow conditions, fecal indicator bacteria counts at the most downstream (integrator) sites in each watershed were considerably higher than counts at those two sites during low-flow conditions. When stormflow sample counts are included with low-flow sample counts to compute a geometric mean for each site, classification changes from fully supporting to not fully supporting contact recreation on the basis of the geometric mean contact

  13. Evaluation of bacteriocinogenic activity, safety traits and biotechnological potential of fecal lactic acid bacteria (LAB), isolated from Griffon Vultures (Gyps fulvus subsp. fulvus).

    PubMed

    Arbulu, Sara; Jiménez, Juan J; Gútiez, Loreto; Campanero, Cristina; Del Campo, Rosa; Cintas, Luis M; Herranz, Carmen; Hernández, Pablo E

    2016-09-29

    Lactic acid bacteria (LAB) are part of the gut microbiota and produce ribosomally synthesized antimicrobial peptides or bacteriocins with interest as natural food preservatives and therapeutic agents. Bacteriocin-producing LAB are also attractive as probiotics. Griffon vultures (Gyps fulvus subspecies fulvus) are scavenger birds that feed almost exclusively on carrion without suffering apparent ill effects. Therefore, griffon vultures might be considered a reservoir of bacteriocin-producing lactic acid bacteria (LAB) with potential biotechnological applications. Griffon vulture feces were screened for LAB with antimicrobial activity, genes encoding bacteriocins, potential virulence determinants, susceptibility to antibiotics, genotyping and characterization of bacteriocins. In this study, from 924 LAB evaluated 332 isolates (36 %) showed direct antimicrobial activity against Gram-positive bacteria only. The molecular identification of the most antagonistic 95 isolates showed that enterococci was the largest LAB group with antimicrobial activity (91 %) and E. faecium (40 %) the most identified antagonistic species. The evaluation of the presence of bacteriocin structural genes in 28 LAB isolates with the highest bacteriocinogenic activity in their supernatants determined that most enterococcal isolates (75 %) encoded multiple bacteriocins, being enterocin A (EntA) the largest identified (46 %) bacteriocin. Most enterococci (88 %) were resistant to multiple antibiotics. ERIC-PCR and MLST techniques permitted genotyping and recognition of the potential safety of the bacteriocinogenic enterococci. A multiple-step chromatographic procedure, determination of the N-terminal amino acid sequence of purified bacteriocins by Edman degradation and a MALDI TOF/TOF tandem MS procedure permitted characterization of bacteriocins present in supernatants of producer cells. Enterococci was the largest LAB group with bacteriocinogenic activity isolated from griffon vulture feces

  14. Methods of targeting animal sources of fecal pollution in water

    EPA Science Inventory

    In this chapter, proposed chemical and biological MST indicators for the determination of animal fecal sources are discussed. The biological indicators are grouped based on the phylogenetic description of the proposed target (eukarya, bacteria, and virus). A comprehensive descrip...

  15. Methods of targeting animal sources of fecal pollution in water

    EPA Science Inventory

    In this chapter, proposed chemical and biological MST indicators for the determination of animal fecal sources are discussed. The biological indicators are grouped based on the phylogenetic description of the proposed target (eukarya, bacteria, and virus). A comprehensive descrip...

  16. Health burden of gastrointestinal symptoms resulting from swimming in fecally-contaminated recreational waters.

    EPA Science Inventory

    Millions of people swim and recreate in oceans and lakes every year. Fecal contamination of these waters can occur from sewage discharges, runoff, and other point and non-point sources. Measures of fecal indicator contamination (e.g., the fecal indicator bacteria E. coli and Ent...

  17. The modified SWAT model for predicting fecal coliform in the Wachusett Reservoir Watershed, USA

    USDA-ARS?s Scientific Manuscript database

    Fecal contamination has been an issue for water quality because fecal coliform bacteria are used as an indicator organism to detect pathogens in water. In order to assess fecal contamination in the Wachusett Reservoir Watershed in Massachusetts, USA, the Soil and Water Assessment Tool (SWAT), a comm...

  18. Monitoring Fecal Indicators and Pathogens in Watersheds: Implementing a Quantitative Microbial Risk Assessment Approach

    EPA Science Inventory

    - Many of the nation's rivers, lakes, and estuaries are impaired with fecal indicator bacteria. - Fecal contamination from point and non-point sources is responsible for the presence of fecal pathogens in source and recreational waters - Effective compliance with TMDL regulatio...

  19. Monitoring Fecal Indicators and Pathogens in Watersheds: Implementing a Quantitative Microbial Risk Assessment Approach

    EPA Science Inventory

    - Many of the nation's rivers, lakes, and estuaries are impaired with fecal indicator bacteria. - Fecal contamination from point and non-point sources is responsible for the presence of fecal pathogens in source and recreational waters - Effective compliance with TMDL regulatio...

  20. Health burden of gastrointestinal symptoms resulting from swimming in fecally-contaminated recreational waters.

    EPA Science Inventory

    Millions of people swim and recreate in oceans and lakes every year. Fecal contamination of these waters can occur from sewage discharges, runoff, and other point and non-point sources. Measures of fecal indicator contamination (e.g., the fecal indicator bacteria E. coli and Ent...

  1. Human and Animal Fecal Contamination of Community Water Sources, Stored Drinking Water and Hands in Rural India Measured with Validated Microbial Source Tracking Assays.

    PubMed

    Schriewer, Alexander; Odagiri, Mitsunori; Wuertz, Stefan; Misra, Pravas R; Panigrahi, Pinaki; Clasen, Thomas; Jenkins, Marion W

    2015-09-01

    We examined pathways of exposure to fecal contamination of human and animal origin in 24 villages in Odisha, India. In a cross-sectional study during the monsoon season, fecal exposure via community water sources (N = 123) and in the home (N = 137) was assessed using human- and nonhuman-associated Bacteroidales microbial source tracking (MST) markers and fecal coliforms (FCs). Detection rates and marker concentrations were examined to pinpoint pathways of human fecal exposure in the public and domestic domains of disease transmission in study communities. Human fecal markers were detected much more frequently in the domestic domain (45% of households) than in public domain sources (8% of ponds; 4% of groundwater drinking sources). Animal fecal markers were widely detected in both domains (74% of ponds, 96% of households, 10% of groundwater drinking sources), indicating ubiquitous risks of exposure to animal feces and zoonotic pathogens. This study confirms an often suggested contamination link from hands to stored water in the home in developing countries separately for mothers' and children's hands and both human and animal fecal contamination. In contrast to MST markers, FCs provided a poor metric to assess risks of exposure to fecal contamination of human origin in this rural setting.

  2. Human and Animal Fecal Contamination of Community Water Sources, Stored Drinking Water and Hands in Rural India Measured with Validated Microbial Source Tracking Assays

    PubMed Central

    Schriewer, Alexander; Odagiri, Mitsunori; Wuertz, Stefan; Misra, Pravas R.; Panigrahi, Pinaki; Clasen, Thomas; Jenkins, Marion W.

    2015-01-01

    We examined pathways of exposure to fecal contamination of human and animal origin in 24 villages in Odisha, India. In a cross-sectional study during the monsoon season, fecal exposure via community water sources (N = 123) and in the home (N = 137) was assessed using human- and nonhuman-associated Bacteroidales microbial source tracking (MST) markers and fecal coliforms (FCs). Detection rates and marker concentrations were examined to pinpoint pathways of human fecal exposure in the public and domestic domains of disease transmission in study communities. Human fecal markers were detected much more frequently in the domestic domain (45% of households) than in public domain sources (8% of ponds; 4% of groundwater drinking sources). Animal fecal markers were widely detected in both domains (74% of ponds, 96% of households, 10% of groundwater drinking sources), indicating ubiquitous risks of exposure to animal feces and zoonotic pathogens. This study confirms an often suggested contamination link from hands to stored water in the home in developing countries separately for mothers' and children's hands and both human and animal fecal contamination. In contrast to MST markers, FCs provided a poor metric to assess risks of exposure to fecal contamination of human origin in this rural setting. PMID:26149868

  3. Recreational water exposures and health effects at a tropical and a runoff impacted beach

    EPA Science Inventory

    Background: Studies conducted by the EPA at beaches with nearby treated sewage discharges established associations between gastrointestinal (GI) illnesses among swimmers and measurements of fecal indicator bacteria, Enterococcus and Bacteroidales (marine beaches only) measured by...

  4. Recreational water exposures and health effects at a tropical and a runoff impacted beach

    EPA Science Inventory

    Background: Studies conducted by the EPA at beaches with nearby treated sewage discharges established associations between gastrointestinal (GI) illnesses among swimmers and measurements of fecal indicator bacteria, Enterococcus and Bacteroidales (marine beaches only) measured by...

  5. Bacteroidales Secreted Antimicrobial Proteins Target Surface Molecules Necessary for Gut Colonization and Mediate Competition In Vivo

    PubMed Central

    Roelofs, Kevin G.; Coyne, Michael J.; Gentyala, Rahul R.; Chatzidaki-Livanis, Maria

    2016-01-01

    ABSTRACT We recently showed that human gut Bacteroidales species secrete antimicrobial proteins (BSAPs), and we characterized in vitro the first such BSAP produced by Bacteroides fragilis. In this study, we identified a second potent BSAP produced by the ubiquitous and abundant human gut species Bacteroides uniformis. The two BSAPs contain a membrane attack complex/perforin (MACPF) domain but share very little sequence similarity. We identified the target molecules of BSAP-sensitive cells and showed that each BSAP targets a different class of surface molecule: BSAP-1 targets an outer membrane protein of sensitive B. fragilis strains, and BSAP-2 targets the O-antigen glycan of lipopolysaccharide (LPS) of sensitive B. uniformis strains. Species-wide genomic and phenotypic analyses of B. fragilis and B. uniformis showed that BSAP-producing strains circumvent killing by synthesizing an orthologous nontargeted surface molecule. The BSAP genes are adjacent to the gene(s) encoding their target replacements, suggesting coacquisition. Using a gnotobiotic mouse competitive-colonization model, we found that the BSAP surface targets are important for colonization of the mammalian gut, thereby explaining why they are maintained in sensitive strains and why they were replaced rather than deleted in BSAP-producing strains. Using isogenic BSAP-producing, -sensitive, and -resistant strains, we show that a BSAP-producing strain outcompetes a sensitive strain but not a resistant strain in the mammalian gut. Human gut metagenomic datasets reveal that BSAP-1-sensitive strains do not cooccur with BSAP-1-producing strains in human gut microbiotas, further supporting the idea that BSAPs are important competitive factors with relevance to the strain-level composition of the human gut microbiota. PMID:27555309

  6. Linking land-use type and stream water quality using spatial data of fecal indicator bacteria and heavy metals in the Yeongsan river basin.

    PubMed

    Kang, Joo-Hyon; Lee, Seung Won; Cho, Kyung Hwa; Ki, Seo Jin; Cha, Sung Min; Kim, Joon Ha

    2010-07-01

    This study reveals land-use factors that explain stream water quality during wet and dry weather conditions in a large river basin using two different linear models-multiple linear regression (MLR) models and constrained least squares (CLS) models. Six land-use types and three topographical parameters (size, slope, and permeability) of the watershed were incorporated into the models as explanatory variables. The suggested models were then demonstrated using a digitized elevation map in conjunction with the land-use and the measured concentration data for Escherichia coli (EC), Enterococci bacteria (ENT), and six heavy metal species collected monthly during 2007-2008 at 50 monitoring sites in the Yeongsan Watershed, Korea. The results showed that the MLR models can be a powerful tool for predicting the average concentrations of pollutants in stream water (the Nash-Sutcliffe (NS) model efficiency coefficients ranged from 0.67 to 0.95). On the other hand, the CLS models, with moderately good prediction performance (the NS coefficients ranged 0.28-0.85), were more suitable for quantifying contributions of respective land-uses to the stream water quality. The CLS models suggested that industrial and urban land-uses are major contributors to the stream concentrations of EC and ENT, whereas agricultural, industrial, and mining areas were significant sources of many heavy metal species. In addition, the slope, size, and permeability of the watershed were found to be important factors determining the extent of the contribution from each land-use type to the stream water quality. The models proposed in this paper can be considered useful tools for developing land cover guidelines and for prioritizing locations for implementing management practices to maintain stream water quality standard in a large river basin.

  7. Factors Influencing Fecal Contamination in Pond of Bangladesh

    NASA Astrophysics Data System (ADS)

    Knappett, P. S.; Escamilla, V.; Layton, A.; McKay, L. D.; Emch, M.; Mailloux, B. J.; Williams, D. E.; Huq, M. R.; Alam, M.; Farhana, L.; Ferguson, A. S.; Sayler, G. S.; Ahmed, K.; Serre, M. L.; Akita, Y.; Yunus, M.; van Geen, A.

    2010-12-01

    Occurrence of diarrheal disease in villages in rural Bangladesh remains relatively common, even though many households have switched to tubewell water for drinking and cooking. One factor contributing to this may be exposure to fecal contamination in ponds, which are often used for bathing and fishing. The objective of this study is to determine the dominant sources of fecal pollution in typical ponds and to explore the relationship between local population, latrine density, latrine quality and concentrations of fecal bacteria and pathogens in pond water. Forty-three ponds were sampled and analyzed for E. coli using culture-based methods and for E. coli, Bacteroides and adenovirus using quantitative PCR. Population and sanitation infrastructure were surveyed and compared to levels of pond fecal contamination. Molecular fecal source tracking using Bacteroides, determined that humans were the dominant source of fecal contamination in 79% of the ponds. Ponds directly receiving latrine effluent had the highest concentrations of fecal indicator bacteria. Concentrations of fecal indicator bacteria correlated with population surveyed within a distance of 30-70 m (p<0.01) and total latrines surveyed within 50-70 m (p<0.05). Unsanitary latrines with visible effluent within the pond drainage basin were also significantly correlated to fecal indicator concentrations (p<0.05). The vast majority of the surveyed ponds contained unsafe levels of fecal contamination primarily due to unsanitary latrines, and to lesser extent to sanitary latrines and cattle. Since the majority of fecal pollution is from humans, use of pond water could help explain the persistence of diarrheal disease in rural Bangladesh.

  8. Evaluating the operational utility of a Bacteroidales quantitative PCR-based MST approach in determining the source of faecal indicator organisms at a UK bathing water.

    PubMed

    Stapleton, Carl M; Kay, David; Wyer, Mark D; Davies, Cheryl; Watkins, John; Kay, Chris; McDonald, Adrian T; Porter, Jonathan; Gawler, Andrew

    2009-11-01

    Microbial source tracking techniques are used in the UK to provide an evidence-base to guide major expenditure decisions and/or regulatory action relating to sewage disposal. Consequently, it is imperative that the techniques used robustly index faecal indicator organisms (FIOs) that are the regulatory parameters for bathing and shellfish harvesting areas. This study reports a 'field-scale' test of microbial source tracking (MST) based on the quantitative PCR analyses of Bacteroidales 16S rRNA genetic marker sequences. The project acquired data to test the operational utility of quantitative Bacteroidales MST data, comparing it with FIO concentrations in streams, effluents and bathing waters. Overall, the data did not exhibit a consistent pattern of significant correlations between Bacteroidales MST parameters and FIOs within the different sample matrices (i.e. rivers, bathing waters and/or effluents). Consequently, there was little evidence from this study that reported concentrations and/or percentages of human and/or ruminant faecal loadings (that are based on Bacteroidales MST gene copy numbers) offer a credible evidence-base describing FIO contributions to receiving water 'non-compliance'. The study also showed (i) there was no significant attenuation of the Bacteroidales gene copy number 'signal' through the UV disinfection process; and (ii) single non-compliant samples submitted for Bacteroidales MST analysis, do not reliably characterise the balance of faecal loadings due to the high variability in the MST signal observed. At this stage in the development of the MST tool deployed, it would be imprudent to use the percentage human and/or ruminant contributions (i.e. as indicated by MST data acquired at a bathing water) as the sole or principal element in the evidence-base used to guide major expenditure decisions and/or regulatory action.

  9. Water quality, weather and environmental factors associated with fecal indicator organism density in beach sand at two recreational marine beaches.

    PubMed

    Heaney, Christopher D; Exum, Natalie G; Dufour, Alfred P; Brenner, Kristen P; Haugland, Richard A; Chern, Eunice; Schwab, Kellogg J; Love, David C; Serre, Marc L; Noble, Rachel; Wade, Timothy J

    2014-11-01

    Recent studies showing an association between fecal indicator organisms (FIOs) in sand and gastrointestinal (GI) illness among beachgoers with sand contact have important public health implications because of the large numbers of people who recreate at beaches and engage in sand contact activities. Yet, factors that influence fecal pollution in beach sand remain unclear. During the 2007 National Epidemiological and Environmental Assessment of Recreational (NEEAR) Water Study, sand samples were collected at three locations (60 m apart) on weekend days (Sat, Sun) and holidays between June and September at two marine beaches - Fairhope Beach, AL and Goddard Beach, RI - with nearby publicly-owned treatment works (POTWs) outfalls. F(+) coliphage, enterococci, Bacteroidales, fecal Bacteroides spp., and Clostridium spp. were measured in sand using culture and qPCR-based calibrator-cell equivalent methods. Water samples were also collected on the same days, times and transects as the 144 sand samples and were assayed using the same FIO measurements. Weather and environmental data were collected at the time of sample collection. Mean FIO concentrations in sand varied over time, but not space. Enterococci CFU and CCE densities in sand were not correlated, although other FIOs in sand were. The strongest correlation between FIO density in sand and water was fecal Bacteroides CCE, followed by enterococci CFU, Clostridium spp. CCE, and Bacteroidales CCE. Overall, the factors associated with FIO concentrations in sand were related to the sand-water interface (i.e., sand-wetting) and included daily average densities of FIOs in water, rainfall, and wave height. Targeted monitoring that focuses on daily trends of sand FIO variability, combined with information about specific water quality, weather, and environmental factors may inform beach monitoring and management decisions to reduce microbial burdens in beach sand. The views expressed in this paper are those of the authors and do

  10. Water quality, weather and environmental factors associated with fecal indicator organism density in beach sand at two recreational marine beaches

    PubMed Central

    Heaney, Christopher D.; Exum, Natalie G.; Dufour, Alfred P.; Brenner, Kristen P.; Haugland, Richard A.; Chern, Eunice; Schwab, Kellogg J.; Love, David C.; Serre, Marc L.; Noble, Rachel; Wade, Timothy J.

    2015-01-01

    Recent studies showing an association between fecal indicator organisms (FIOs) in sand and gastrointestinal (GI) illness among beachgoers with sand contact have important public health implications because of the large numbers of people who recreate at beaches and engage in sand contact activities. Yet, factors that influence fecal pollution in beach sand remain unclear. During the 2007 National Epidemiological and Environmental Assessment of Recreational (NEEAR) Water Study, sand samples were collected at three locations (60 m apart) on weekend days (Sat, Sun) and holidays between June and September at two marine beaches — Fairhope Beach, AL and Goddard Beach, RI — with nearby publicly-owned treatment works (POTWs) outfalls. F+ coliphage, enterococci, Bacteroidales, fecal Bacteroides spp., and Clostridium spp. were measured in sand using culture and qPCR-based calibrator-cell equivalent methods. Water samples were also collected on the same days, times and transects as the 144 sand samples and were assayed using the same FIO measurements. Weather and environmental data were collected at the time of sample collection. Mean FIO concentrations in sand varied over time, but not space. Enterococci CFU and CCE densities in sand were not correlated, although other FIOs in sand were. The strongest correlation between FIO density in sand and water was fecal Bacteroides CCE, followed by enterococci CFU, Clostridium spp. CCE, and Bacteroidales CCE. Overall, the factors associated with FIO concentrations in sand were related to the sand–water interface (i.e., sand-wetting) and included daily average densities of FIOs in water, rainfall, and wave height. Targeted monitoring that focuses on daily trends of sand FIO variability, combined with information about specific water quality, weather, and environmental factors may inform beach monitoring and management decisions to reduce microbial burdens in beach sand. The views expressed in this paper are those of the authors

  11. Water quality indicators and the risk of illness at beaches with nonpoint sources of fecal contaminants

    EPA Science Inventory

    BACKGROUND: Indicator bacteria are a good predictor of illness at marine beaches that have point sources of pollution with human fecal content. Few studies have addressed the utility of indicator bacteria where nonpoint sources are the dominant fecal input. Extrapolating current ...

  12. Water quality indicators and the risk of illness at beaches with nonpoint sources of fecal contaminants

    EPA Science Inventory

    BACKGROUND: Indicator bacteria are a good predictor of illness at marine beaches that have point sources of pollution with human fecal content. Few studies have addressed the utility of indicator bacteria where nonpoint sources are the dominant fecal input. Extrapolating current ...

  13. Fecal Pollution of Water.

    EPA Science Inventory