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...

Bacterial indicator occurrence and the use of an F+ specific RNA coliphage assay to identify fecal sources in Homosassa Springs, Florida

USGS Publications Warehouse

Griffin, Dale W.; Stokes, Rodger; Rose, J.B.; Paul, J.H.

2000-01-01

A microbiological water quality study of Homosassa Springs State Wildlife Park (HSSWP) and surrounding areas was undertaken. Samples were collected in November of 1997 (seven sites) and again in November of 1998 (nine sites). Fecal bacterial concentrations (total and fecal coliforms, Clostridium perfringens, and enterococci) were measured as relative indicators of fecal contamination. F+-specific coliphage genotyping was performed to determine the source of fecal contamination at the study sites. Bacterial levels were considerably higher at most sites in the 1997 sampling compared to the 1998 sampling, probably because of the greater rainfall that year. In November of 1997, 2 of the 7 sites were in violation of all indicator standards and guidance levels. In November of 1998, 1 of 9 sites was in violation of all indicator standard and guidance levels. The highest concentrations of all fecal indicators were found at a station downstream of the animal holding pens in HSSWP. The lowest levels of indicators were found at the Homosassa Main Spring vent. Levels of fecal indicators downstream of HSSWP (near the point of confluence with the river) were equivalent to those found in the Southeastern Fork and areas upstream of the park influences. F+ specific RNA coliphage analysis indicated that fecal contamination at all sites that tested positive was from animal sources (mammals and birds). These results suggest that animal (indigenous and those in HSSWP) and not human sources influenced microbial water quality in the area of Homosassa River covered by this study.

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...

Origin of fecal contamination in waters from contrasted areas: stanols as Microbial Source Tracking markers.

PubMed

Derrien, M; Jardé, E; Gruau, G; Pourcher, A M; Gourmelon, M; Jadas-Hécart, A; Pierson Wickmann, A C

2012-09-01

Improving the microbiological quality of coastal and river waters relies on the development of reliable markers that are capable of determining sources of fecal pollution. Recently, a principal component analysis (PCA) method based on six stanol compounds (i.e. 5β-cholestan-3β-ol (coprostanol), 5β-cholestan-3α-ol (epicoprostanol), 24-methyl-5α-cholestan-3β-ol (campestanol), 24-ethyl-5α-cholestan-3β-ol (sitostanol), 24-ethyl-5β-cholestan-3β-ol (24-ethylcoprostanol) and 24-ethyl-5β-cholestan-3α-ol (24-ethylepicoprostanol)) was shown to be suitable for distinguishing between porcine and bovine feces. In this study, we tested if this PCA method, using the above six stanols, could be used as a tool in "Microbial Source Tracking (MST)" methods in water from areas of intensive agriculture where diffuse fecal contamination is often marked by the co-existence of human and animal sources. In particular, well-defined and stable clusters were found in PCA score plots clustering samples of "pure" human, bovine and porcine feces along with runoff and diluted waters in which the source of contamination is known. A good consistency was also observed between the source assignments made by the 6-stanol-based PCA method and the microbial markers for river waters contaminated by fecal matter of unknown origin. More generally, the tests conducted in this study argue for the addition of the PCA method based on six stanols in the MST toolbox to help identify fecal contamination sources. The data presented in this study show that this addition would improve the determination of fecal contamination sources when the contamination levels are low to moderate. Copyright © 2012 Elsevier Ltd. All rights reserved.

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

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.

Stanols as a tool to track the origin of microbial contamination of oysters, Crassostrea gigas, in shellfish areas.

NASA Astrophysics Data System (ADS)

Harrault, Loïc; Jardé, Emilie; Jeanneau, Laurent; Petitjean, Patrice

2013-04-01

Runoff of cattle manures (cows, pigs, sheeps) or discharge of effluent from wastewater treatment plants (WWTP) into aquatic ecosystems can lead to microbiological contamination of waters and living organisms. In coastal ecosystems and particularly in shellfish harvesting areas, the presence of pathogen microorganisms in waters induces fecal contamination of filter feeding bivalves (oysters, mussels, scallops…), therefore leading to human health risks associated to the consumption of these contaminated organisms. Watershed management plans that aim at limiting these risks require the development of tools able to identify fecal contamination sources. The fecal indicator bacteria used in the regulations to determine fecal contamination are not source specific since they are found in the feces of most warm-blooded animals. Thus, microbiological biomarkers have been developed in association with chemical biomarkers as Microbial Source Tracking (MST) methods. Fecal stanols, by-products of sterols obtained by human and animal microbial gut flora, are found in considerable amounts in feces with different relative proportions depending on their animal or human source. Recently, in association with microbiological biomarkers, the stanol fingerprint of contaminated waters has been successfully used to determine the main source of fecal contamination (cow, pig or human sources) in rural watersheds (Brittany, France). Up to now, the use of the stanol fingerprint to track the fecal contamination in shellfish tissues, especially bivalves, has been limited to the analysis of coprostanol, a stanol commonly associated to human contamination. Therefore, whether the stanol fingerprint can be used as a MST method in bivalves or not is still unknown. The first aim of this study was to compare several organic extraction procedures of stanols in the oyster Crassostrea gigas to determine a reliable method for stanol fingerprint analysis in bivalves. Solvent extraction and purification steps have been carried out with attention as they are critical for stanol quantification. Secondly, the evolution of the stanol fingerprint of oysters with time was evaluated during 6 days by artificially contaminating microcosms with two concentrations of a WWTP effluent. In the microcosms, the fingerprint of stanols as a chemical biomarkers of fecal (human) contamination was compared to counts of Escherichia coli, a commonly used microbial indicator. In association with microbial markers, the method developed from the two previous steps will be applied at the watershed scale in order to identify sources of fecal contamination in Brittany and Normandy (France).

Extent of Fecal Contamination of Household Drinking Water in Nepal: Further Analysis of Nepal Multiple Indicator Cluster Survey 2014.

PubMed

Kandel, Pragya; Kunwar, Ritu; Lamichhane, Prabhat; Karki, Surendra

2017-02-08

Water sources classified as "improved" may not necessarily provide safe drinking water for householders. We analyzed data from Nepal Multiple Indicator Cluster Survey 2014 to explore the extent of fecal contamination of household drinking water. Fecal contamination was detected in 81.2% (95% confidence interval [CI]: 77.9-84.2) household drinking water from improved sources and 89.6% (95% CI: 80.4-94.7) in water samples from unimproved sources. In adjusted analysis, there was no difference in odds of fecal contamination of household drinking water between improved and unimproved sources. We observed significantly lower odds of fecal contamination of drinking water in households in higher wealth quintiles, where soap and water were available for handwashing and in households employing water treatment. The extent of contamination of drinking water as observed in this study highlights the huge amount of effort required to ensure the provision of safely managed water in Nepal by 2030 as aimed in sustainable development goals. © The American Society of Tropical Medicine and Hygiene.

Extent of Fecal Contamination of Household Drinking Water in Nepal: Further Analysis of Nepal Multiple Indicator Cluster Survey 2014

PubMed Central

Kandel, Pragya; Kunwar, Ritu; Lamichhane, Prabhat; Karki, Surendra

2017-01-01

Water sources classified as “improved” may not necessarily provide safe drinking water for householders. We analyzed data from Nepal Multiple Indicator Cluster Survey 2014 to explore the extent of fecal contamination of household drinking water. Fecal contamination was detected in 81.2% (95% confidence interval [CI]: 77.9–84.2) household drinking water from improved sources and 89.6% (95% CI: 80.4–94.7) in water samples from unimproved sources. In adjusted analysis, there was no difference in odds of fecal contamination of household drinking water between improved and unimproved sources. We observed significantly lower odds of fecal contamination of drinking water in households in higher wealth quintiles, where soap and water were available for handwashing and in households employing water treatment. The extent of contamination of drinking water as observed in this study highlights the huge amount of effort required to ensure the provision of safely managed water in Nepal by 2030 as aimed in sustainable development goals. PMID:27821687

Differentiation of fecal Escherichia coli from poultry and free-living birds by (GTG)5-PCR genomic fingerprinting.

PubMed

Mohapatra, Bidyut R; Broersma, Klaas; Mazumder, Asit

2008-04-01

Determination of the non-point sources of fecal pollution is essential for the assessment of potential public health risk and development of appropriate management practices for prevention of further contamination. Repetitive extragenic palindromic-PCR coupled with (GTG)(5) primer [(GTG)(5)-PCR] was performed on 573 Escherichia coli isolates obtained from the feces of poultry (chicken, duck and turkey) and free-living (Canada goose, hawk, magpie, seagull and songbird) birds to evaluate the efficacy of (GTG)(5)-PCR genomic fingerprinting in the prediction of the correct source of fecal pollution. A discriminant analysis with the jack-knife algorithm of (GTG)(5)-PCR DNA fingerprints revealed that 95%, 94.1%, 93.2%, 84.6%, 79.7%, 76.7%, 75.3% and 70.7% of magpie, hawk, turkey, seagull, Canada goose, chicken, duck and songbird fecal E. coli isolates classified into the correct host source, respectively. The results of this study indicate that (GTG)(5)-PCR can be considered to be a complementary molecular tool for the rapid determination of E. coli isolates identity and tracking the non-point sources of fecal pollution.

Differential Decay of Enterococci and Escherichia coli Originating from Two Fecal Pollution Sources

PubMed Central

Harwood, Valerie J.; Shanks, Orin C.; Fout, G. Shay; Ashbolt, Nicholas J.

2013-01-01

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 habitats. In this study, sunlight exposure and indigenous aquatic microbiota were also important contributors, whose effects on FIB also differed between water types. PMID:23377944

Molecular characterization of bacteriophages for microbial source tracking in Korea.

PubMed

Lee, Jung Eun; Lim, Mi Young; Kim, Sei Yoon; Lee, Sunghee; Lee, Heetae; Oh, Hyun-Myung; Hur, Hor-Gil; Ko, Gwangpyo

2009-11-01

We investigated coliphages from various fecal sources, including humans and animals, for microbial source tracking in South Korea. Both somatic and F+-specific coliphages were isolated from 43 fecal samples from farms, wild animal habitats, and human wastewater plants. Somatic coliphages were more prevalent and abundant than F+ coliphages in all of the tested fecal samples. We further characterized 311 F+ coliphage isolates using RNase sensitivity assays, PCR and reverse transcription-PCR, and nucleic acid sequencing. Phylogenetic analyses were performed based on the partial nucleic acid sequences of 311 F+ coliphages from various sources. F+ RNA coliphages were most prevalent among geese (95%) and were least prevalent in cows (5%). Among the genogroups of F+ RNA coliphages, most F+ coliphages isolated from animal fecal sources belonged to either group I or group IV, and most from human wastewater sources were in group II or III. Some of the group I coliphages were present in both human and animal source samples. F+ RNA coliphages isolated from various sources were divided into two main clusters. All F+ RNA coliphages isolated from human wastewater were grouped with Qbeta-like phages, while phages isolated from most animal sources were grouped with MS2-like phages. UniFrac significance statistical analyses revealed significant differences between human and animal bacteriophages. In the principal coordinate analysis (PCoA), F+ RNA coliphages isolated from human waste were distinctively separate from those isolated from other animal sources. However, F+ DNA coliphages were not significantly different or separate in the PCoA. These results demonstrate that proper analysis of F+ RNA coliphages can effectively distinguish fecal sources.

COMPARISON OF THE TEMPORAL VARIABILITY OF ENTEROCOCCAL CLUSTERS IN IMPACTED STREAMS USING A MULTIPLEX POLYMERASE CHAIN REACTION PROCEDURE

EPA Science Inventory

Understanding how fecal indicator bacteria and/or fecal indicator genotypes vary over time is important to determine the sources of fecal contamination. Enterococcus is one of the two indicators recommended by the EPA to monitor freshwaters for fecal contamination. Along with E...

Comparison of bacteroides-prevotella 16S rRNA genetic markers for fecal samples from different animal species.

PubMed

Fogarty, Lisa R; Voytek, Mary A

2005-10-01

To effectively manage surface and ground waters it is necessary to improve our ability to detect and identify sources of fecal contamination. We evaluated the use of the anaerobic bacterial group Bacteroides-Prevotella as a potential fecal indicator. Terminal restriction length polymorphism (T-RFLP) of the 16S rRNA genes from this group was used to determine differences in populations and to identify any unique populations in chickens, cows, deer, dogs, geese, horses, humans, pigs, and seagulls. The group appears to be a good potential fecal indicator in all groups tested except for avians. Cluster analysis of Bacteroides-Prevotella community T-RFLP profiles indicates that Bacteroides-Prevotella populations from samples of the same host species are much more similar to each other than to samples from different source species. We were unable to identify unique peaks that were exclusive to any source species; however, for most host species, at least one T-RFLP peak was identified to be more commonly found in that species, and a combination of peaks could be used to identify the source. T-RFLP profiles obtained from water spiked with known-source feces contained the expected diagnostic peaks from the source. These results indicate that the approach of identifying Bacteroides-Prevotella molecular markers associated with host species might be useful in identifying sources of fecal contamination in the environment.

Comparison of Bacteroides-Prevotella 16S rRNA Genetic Markers for Fecal Samples from Different Animal Species

PubMed Central

Fogarty, Lisa R.; Voytek, Mary A.

2005-01-01

To effectively manage surface and ground waters it is necessary to improve our ability to detect and identify sources of fecal contamination. We evaluated the use of the anaerobic bacterial group Bacteroides-Prevotella as a potential fecal indicator. Terminal restriction length polymorphism (T-RFLP) of the 16S rRNA genes from this group was used to determine differences in populations and to identify any unique populations in chickens, cows, deer, dogs, geese, horses, humans, pigs, and seagulls. The group appears to be a good potential fecal indicator in all groups tested except for avians. Cluster analysis of Bacteroides-Prevotella community T-RFLP profiles indicates that Bacteroides-Prevotella populations from samples of the same host species are much more similar to each other than to samples from different source species. We were unable to identify unique peaks that were exclusive to any source species; however, for most host species, at least one T-RFLP peak was identified to be more commonly found in that species, and a combination of peaks could be used to identify the source. T-RFLP profiles obtained from water spiked with known-source feces contained the expected diagnostic peaks from the source. These results indicate that the approach of identifying Bacteroides-Prevotella molecular markers associated with host species might be useful in identifying sources of fecal contamination in the environment. PMID:16204514

Comparison of Bacteroides-Prevotella 16S rRNA genetic markers for fecal samples from different animal species

USGS Publications Warehouse

Fogarty, L.R.; Voytek, M.A.

2005-01-01

To effectively manage surface and ground waters it is necessary to improve our ability to detect and identify sources of fecal contamination. We evaluated the use of the anaerobic bacterial group Bacteroides-Prevotella as a potential fecal indicator. Terminal restriction length polymorphism (T-RFLP) of the 16S rRNA genes from this group was used to determine differences in populations and to identify any unique populations in chickens, cows, deer, dogs, geese, horses, humans, pigs, and seagulls. The group appears to be a good potential fecal indicator in all groups tested except for avians. Cluster analysis of Bacteroides-Prevotella community T-RFLP profiles indicates that Bacteroides-Prevotella populations from samples of the same host species are much more similar to each other than to samples from different source species. We were unable to identify unique peaks that were exclusive to any source species; however, for most host species, at least one T-RFLP peak was identified to be more commonly found in that species, and a combination of peaks could be used to identify the source. T-RFLP profiles obtained from water spiked with known-source feces contained the expected diagnostic peaks from the source. These results indicate that the approach of identifying Bacteroides-Prevotella molecular markers associated with host species might be useful in identifying sources of fecal contamination in the environment.

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

Assessment of possible sources of microbiological contamination in the water column and streambed sediment of the Jacks Fork, Ozark National Scenic Riverways, Missouri - Phase III

USGS Publications Warehouse

Davis, Jerri V.; Barr, Miya N.

2006-01-01

In 1998, a 5 river-mile reach of the Jacks Fork was included on Missouri's list of impaired waters as required by Section 303(d) of the Federal Clean Water Act. The identified pollutant on the Jacks Fork was fecal coliform bacteria. The length of the impaired reach was changed to 7 miles on the Missouri 2002 303(d) list because of data indicating the fecal coliform bacteria problem existed over a broader area. The U.S. Geological Survey, in cooperation with the National Park Service, conducted a study to better understand the extent and sources of microbiological contamination within the Jacks Fork from Alley Spring to the mouth, which includes the 7-mile 303(d) reach. Ten sites were sampled from June 2003 through October 2003 and from June 2004 through October 2004. Water-column and streambed sediment samples were collected from main-stem and tributary sites mostly during base-flow conditions during a variety of recreational season river uses and analyzed for fecal coliform and Escherichia coli bacteria. Isolates of Escherichia coli obtained from water samples collected at five sites were submitted for rep-PCR analysis to identify presumptive sources of fecal indicator bacteria in the Jacks Fork. Results indicate that recreational users (including boaters and swimmers) are not the primary source of fecal coliform bacteria in the Jacks Fork; rather, the presence of fecal coliform bacteria is associated with other animals, of which horses are the primary source. Increases in fecal coliform bacteria densities in the Jacks Fork are associated with cross-country horseback trail-riding events.

Identifying fecal pollution sources using 3M(™) Petrifilm (™) count plates and antibiotic resistance analysis in the Horse Creek Watershed in Aiken County, SC (USA).

PubMed

Harmon, S Michele; West, Ryan T; Yates, James R

2014-12-01

Sources of fecal coliform pollution in a small South Carolina (USA) watershed were identified using inexpensive methods and commonly available equipment. Samples from the upper reaches of the watershed were analyzed with 3M(™) Petrifilm(™) count plates. We were able to narrow down the study's focus to one particular tributary, Sand River, that was the major contributor of the coliform pollution (both fecal and total) to a downstream reservoir that is heavily used for recreation purposes. Concentrations of total coliforms ranged from 2,400 to 120,333 cfu/100 mL, with sharp increases in coliform counts observed in samples taken after rain events. Positive correlations between turbidity and fecal coliform counts suggested a relationship between fecal pollution and stormwater runoff. Antibiotic resistance analysis (ARA) compared antibiotic resistance profiles of fecal coliform isolates from the stream to those of a watershed-specific fecal source library (equine, waterfowl, canines, and untreated sewage). Known fecal source isolates and unknown isolates from the stream were exposed to six antibiotics at three concentrations each. Discriminant analysis grouped known isolates with an overall average rate of correct classification (ARCC) of 84.3 %. A total of 401 isolates from the first stream location were classified as equine (45.9 %), sewage (39.4 %), waterfowl (6.2 %), and feline (8.5 %). A similar pattern was observed at the second sampling location, with 42.6 % equine, 45.2 % sewage, 2.8 % waterfowl, 0.6 % canine, and 8.8 % feline. While there were slight weather-dependent differences, the vast majority of the coliform pollution in this stream appeared to be from two sources, equine and sewage. This information will contribute to better land use decisions and further justify implementation of low-impact development practices within this urban watershed.

SPECIFICITY AND SENSITIVITY OF FECAL BACTEROIDETES HUMAN-SPECIFIC PRIMERS WITH FECAL AND WASTEWATER SAMPLES FROM THE U.S. MIDWEST AND NORTHEAST REGIONS

EPA Science Inventory

Numerous watersheds throughout the United States are impaired due to fecal contamination. Fecal Bacteroidetes is a group of anaerobic bacteria present in high concentrations in animal feces that has shown promise as a microbial source tracking indicator of human and othe...

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...

Characterization of fecal concentrations in human and other animal sources by physical, culture-based, and quantitative real-time PCR methods

EPA Science Inventory

The characteristics of fecal sources, and the ways in which they are measured, can profoundly influence the interpretation of which sources are contaminating water. Although feces from various hosts are known to differ, it is not well understood how those differences compare acro...

USE OF BACTEROIDES PCR-BASED METHODS TO EXAMINE FECAL CONTAMINATION SOURCES IN TROPICAL COASTAL WATERS

EPA Science Inventory

Several library independent Microbial Source Tracking methods have been developed to rapidly determine the source of fecal contamination. Thus far, none of these methods have been tested in tropical marine waters. In this study, we used a Bacteroides 16S rDNA PCR-based...

National Risk Management Research Laboratory (NRMRL) Microbial Research

EPA Science Inventory

Experimental design: Three host-specific PCR assays were tested against fecal and water samples. Host-specificity assays were performed against targeted and nontargeted fecal sources. Detection limits were performed against diluted fecal and water DNA extracts. Groundwater an...

Influence of Coliform Source on Evaluation of Membrane Filters

PubMed Central

Brodsky, M. H.; Schiemann, D. A.

1975-01-01

Four brands of membrane filters were examined for total and fecal coliform recovery performance by two experimental approaches. Using diluted EC broth cultures of water samples, Johns-Manville filters were superior to Sartorius filters for fecal coliform but equivalent for total coliform recovery. Using river water samples, Johns-Manville filters were superior to Sartorius filters for total coliform but equivalent for fecal coliform recovery. No differences were observed between Johns-Manville and Millipore or Millipore and Sartorius filters for total or fecal coliform recoveries using either approach, nor was any difference observed between Millipore and Gelman filters for fecal coliform recovery from river water samples. These results indicate that the source of the coliform bacteria has an important influence on the conclusions of membrane filter evaluation studies. PMID:1106318

Fecal indicator sources in small creeks

USDA-ARS?s Scientific Manuscript database

Fecal indicator organisms (FIO) in surface water have been widely adopted as surrogates for human gastro-intestinal pathogens, despite available data indicating that the correlations are quite low. Nonetheless, high levels of FIO are a source of concern for agricultural, recreational and industrial ...

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...

Assessment of fecal pollution sources in a small northern-plains watershed using PCR and phylogenetic analyses of Bacteroidetes 16S rRNA gene

USGS Publications Warehouse

Lamendella, R.; Domingo, J.W.S.; Oerther, D.B.; Vogel, J.R.; Stoeckel, D.M.

2007-01-01

We evaluated the efficacy, sensitivity, host-specificity, and spatial/temporal dynamics of human- and ruminant-specific 16S rRNA gene Bacteroidetes markers used to assess the sources of fecal pollution in a fecally impacted watershed. Phylogenetic analyses of 1271 fecal and environmental 16S rRNA gene clones were also performed to study the diversity of Bacteroidetes in this watershed. The host-specific assays indicated that ruminant feces were present in 28-54% of the water samples and in all sampling seasons, with increasing frequency in downstream sites. The human-targeted assays indicated that only 3-5% of the water samples were positive for human fecal signals, although a higher percentage of human-associated signals (19-24%) were detected in sediment samples. Phylogenetic analysis indicated that 57% of all water clones clustered with yet-to-be-cultured Bacteroidetes species associated with sequences obtained from ruminant feces, further supporting the prevalence of ruminant contamination in this watershed. However, since several clusters contained sequences from multiple sources, future studies need to consider the potential cosmopolitan nature of these bacterial populations when assessing fecal pollution sources using Bacteroidetes markers. Moreover, additional data is needed in order to understand the distribution of Bacteroidetes host-specific markers and their relationship to water quality regulatory standards. ?? 2006 Federation of European Microbiological Societies.

Male-specific coliphages for source tracking fecal contamination in irrigation waters and prevalence of Shiga toxigenic Escherichia coli in a major produce production region of central coast of California

USDA-ARS?s Scientific Manuscript database

Determining the environmental sources for Shiga toxigenic Escherichia coli is of paramount importance. Since dairy or feedlot cattle are likely sources for this pathogen, determining the sources of fecal contamination may provide supplemental data to traditional trace-back studies from fork to farm....

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...

Water quality of Bear Creek basin, Jackson County, Oregon

USGS Publications Warehouse

Wittenberg, Loren A.; McKenzie, Stuart W.

1980-01-01

Water-quality data identify surface-water-quality problems in Bear Creek basin, Jackson County, Oreg., where possible, their causes or sources. Irrigation and return-flow data show pastures are sources of fecal coliform and fecal streptococci bacteria and sinks for suspended sediment and nitrite-plus-nitrate nitrogen. Bear Creek and its tributaries have dissolved oxygen and pH values that do not meet State standards. Forty to 50% of the fecal coliform and fecal streptococci concentrations were higher than 1,000 bacteria colonies per 100 milliliters during the irrigation season in the lower two-thirds of the basin. During the irrigation season, suspended-sediment concentrations, average 35 milligrams per liter, were double those for the nonirrigation season. The Ashland sewage-treatment plant is a major source of nitrite plus nitrate, ammonia, and Kjeldahl nitrogen, and orthophosphate in Bear Creek. (USGS)

Evaluation of Two PCR-Based Swine-Specific Fecal Source Tracking Assays (Poster)

EPA Science Inventory

Several PCR-based methods have been proposed to identify swine fecal pollution in environmental waters. However, the specificity and distribution of these targets have not been adequately assessed. Consequently, the utility of these assays in identifying swine fecal contamination...

Assessing the sources of high fecal coliform levels at an urban tropical beach

PubMed Central

Davino, Aline Mendonça Cavalcante; de Melo, Milena Bandeira; Caffaro, Roberto Augusto

2015-01-01

Recreational water quality is commonly assessed by microbial indicators such as fecal coliforms. Maceió is the capital of Alagoas state, located in tropical northeastern Brazil. Its beaches are considered as the most beautiful urban beaches in the country. Jatiúca Beach in Maceió was found to be unsuitable for bathing continuously during the year of 2011. The same level of contamination was not observed in surrounding beaches. The aim of this study was to initiate the search for the sources of these high coliform levels, so that contamination can be eventually mitigated. We performed a retrospective analysis of historical results of fecal coliform concentrations from 2006 to 2012 at five monitoring stations located in the study region. Results showed that Jatiúca Beach consistently presented the worst quality among the studied beaches. A field survey was conducted to identify existing point and non-point sources of pollution in the area. Monitoring in the vicinity of Jatiúca was spatially intensified. Fecal coliform concentrations were categorized according to tide range and tide stage. A storm drain located in northern Jatiúca was identified as the main point source of the contamination. However, fecal coliform concentrations at Jatiúca were high during high tides and spring tides even when this point source was inactive (no rainfall). We hypothesize that high fecal coliform levels in Jatiúca Beach may also be caused by aquifer contamination or, more likely, from tide washing of contaminated sand. Both of these hypotheses will be further investigated. PMID:26691459

Assessing the sources of high fecal coliform levels at an urban tropical beach.

PubMed

Davino, Aline Mendonça Cavalcante; de Melo, Milena Bandeira; Caffaro Filho, Roberto Augusto

2015-01-01

Recreational water quality is commonly assessed by microbial indicators such as fecal coliforms. Maceió is the capital of Alagoas state, located in tropical northeastern Brazil. Its beaches are considered as the most beautiful urban beaches in the country. Jatiúca Beach in Maceió was found to be unsuitable for bathing continuously during the year of 2011. The same level of contamination was not observed in surrounding beaches. The aim of this study was to initiate the search for the sources of these high coliform levels, so that contamination can be eventually mitigated. We performed a retrospective analysis of historical results of fecal coliform concentrations from 2006 to 2012 at five monitoring stations located in the study region. Results showed that Jatiúca Beach consistently presented the worst quality among the studied beaches. A field survey was conducted to identify existing point and non-point sources of pollution in the area. Monitoring in the vicinity of Jatiúca was spatially intensified. Fecal coliform concentrations were categorized according to tide range and tide stage. A storm drain located in northern Jatiúca was identified as the main point source of the contamination. However, fecal coliform concentrations at Jatiúca were high during high tides and spring tides even when this point source was inactive (no rainfall). We hypothesize that high fecal coliform levels in Jatiúca Beach may also be caused by aquifer contamination or, more likely, from tide washing of contaminated sand. Both of these hypotheses will be further investigated.

Multi-Disciplinary Approach to Trace Contamination of Streams and Beaches

USGS Publications Warehouse

Nickles, James

2008-01-01

Concentrations of fecal-indicator bacteria in urban streams and ocean beaches in and around Santa Barbara occasionally can exceed public-health standards for recreation. The U.S. Geological Survey (USGS), working with the City of Santa Barbara, has used multi-disciplinary science to trace the sources of the bacteria. This research is helping local agencies take steps to improve recreational water quality. The USGS used an approach that combined traditional hydrologic and microbiological data, with state-of-the-art genetic, molecular, and chemical tracer analysis. This research integrated physical data on streamflow, ground water, and near-shore oceanography, and made extensive use of modern geophysical and isotopic techniques. Using those techniques, the USGS was able to evaluate the movement of water and the exchange of ground water with near-shore ocean water. The USGS has found that most fecal bacteria in the urban streams came from storm-drain discharges, with the highest concentrations occurring during storm flow. During low streamflow, the concentrations varied as much as three-fold, owing to variable contribution of non-point sources such as outdoor water use and urban runoff to streamflow. Fecal indicator bacteria along ocean beaches were from both stream discharge to the ocean and from non-point sources such as bird fecal material that accumulates in kelp and sand at the high-tide line. Low levels of human-specific Bacteroides, suggesting fecal material from a human source, were consistently detected on area beaches. One potential source, a local sewer line buried beneath the beach, was found not to be responsible for the fecal bacteria.

Assessment of Equine Fecal Contamination: The Search for Alternative Bacterial Source-tracking Targets

EPA Science Inventory

16S rDNA clone libraries were evaluated for detection of fecal source-identifying bacteria from a collapsed equine manure pile. Libraries were constructed using universal eubacterial primers and Bacteroides-Prevotella group-specific primers. Eubacterial sequences indicat...

Differential Decay of Bacterial and Viral Fecal Indicators in Common Human Pollution Sources

EPA Science Inventory

Understanding the decomposition of different human fecal pollution sources is necessary for proper implementation of many water quality management practices, as well as predicting associated public health risks. Here, the decay of select cultivated and molecular indicators of fe...

Human-Associated Bacteroides spp. and Human Polyomaviruses as Microbial Source Tracking Markers in Hawaii

PubMed Central

Caffaro-Filho, Roberto A.; Wong, Mayee; Harwood, Valerie J.; Moravcik, Philip; Fujioka, Roger S.

2016-01-01

ABSTRACT Identification of sources of fecal contaminants is needed to (i) determine the health risk associated with recreational water use and (ii) implement appropriate management practices to mitigate this risk and protect the environment. This study evaluated human-associated Bacteroides spp. (HF183TaqMan) and human polyomavirus (HPyV) markers for host sensitivity and specificity using human and animal fecal samples collected in Hawaii. The decay rates of those markers and indicator bacteria were identified in marine and freshwater microcosms exposed and not exposed to sunlight, followed by field testing of the usability of the molecular markers. Both markers were strongly associated with sewage, although the cross-reactivity of the HF183TaqMan (also present in 82% of canine [n = 11], 30% of mongoose [n = 10], and 10% of feline [n = 10] samples) needs to be considered. Concentrations of HF183TaqMan in human fecal samples exceeded those in cross-reactive animals at least 1,000-fold. In the absence of sunlight, the decay rates of both markers were comparable to the die-off rates of enterococci in experimental freshwater and marine water microcosms. However, in sunlight, the decay rates of both markers were significantly lower than the decay rate of enterococci. While both markers have their individual limitations in terms of sensitivity and specificity, these limitations can be mitigated by using both markers simultaneously; ergo, this study supports the concurrent use of HF183TaqMan and HPyV markers for the detection of sewage contamination in coastal and inland waters in Hawaii. IMPORTANCE This study represents an in-depth characterization of microbial source tracking (MST) markers in Hawaii. The distribution and concentrations of HF183TaqMan and HPyV markers in human and animal fecal samples and in wastewater, coupled with decay data obtained from sunlight-exposed and unexposed microcosms, support the concurrent application of HF183TaqMan and HPyV markers for sewage contamination detection in Hawaii waters. Both markers are more conservative and more specific markers of sewage than fecal indicator bacteria (enterococci and Escherichia coli). Analysis of HF183TaqMan (or newer derivatives) is recommended for inclusion in future epidemiological studies concerned with beach water quality, while better concentration techniques are needed for HPyV. Such epidemiological studies can be used to develop new recreational water quality criteria, which will provide direct information on the absence or presence of sewage contamination in water samples as well as reliable measurements of the risk of waterborne disease transmission to swimmers. PMID:27613686

Individual identification of Sitka black-tailed deer (Odocoileus hemionus sitkensis) using DNA from fecal pellets

Treesearch

Todd J. Brinkman; David K. Person; Michael K. Schwartz; Kristine L. Pilgrim; Kevin E. Colson; Kris J. Hundertmark

2010-01-01

We tested a protocol for extracting DNA from fecal pellets from Sitka black-tailed deer (Odocoileus hemionus sitkensis) and evaluated genotyping performance of previously developed microsatellite markers as well as a suite of new markers designed specifically for this study. We screened 30 microsatellites, and identified 7 (23%) loci including 4 new markers, that fit...

Patterns of Antimicrobial Resistance Observed in Escherichia coli Isolates Obtained from Domestic- and Wild-Animal Fecal Samples, Human Septage, and Surface Water

PubMed Central

Sayah, Raida S.; Kaneene, John B.; Johnson, Yvette; Miller, RoseAnn

2005-01-01

A repeated cross-sectional study was conducted to determine the patterns of antimicrobial resistance in 1,286 Escherichia coli strains isolated from human septage, wildlife, domestic animals, farm environments, and surface water in the Red Cedar watershed in Michigan. Isolation and identification of E. coli were done by using enrichment media, selective media, and biochemical tests. Antimicrobial susceptibility testing by the disk diffusion method was conducted for neomycin, gentamicin, streptomycin, chloramphenicol, ofloxacin, trimethoprim-sulfamethoxazole, tetracycline, ampicillin, nalidixic acid, nitrofurantoin, cephalothin, and sulfisoxazole. Resistance to at least one antimicrobial agent was demonstrated in isolates from livestock, companion animals, human septage, wildlife, and surface water. In general, E. coli isolates from domestic species showed resistance to the largest number of antimicrobial agents compared to isolates from human septage, wildlife, and surface water. The agents to which resistance was demonstrated most frequently were tetracycline, cephalothin, sulfisoxazole, and streptomycin. There were similarities in the patterns of resistance in fecal samples and farm environment samples by animal, and the levels of cephalothin-resistant isolates were higher in farm environment samples than in fecal samples. Multidrug resistance was seen in a variety of sources, and the highest levels of multidrug-resistant E. coli were observed for swine fecal samples. The fact that water sample isolates were resistant only to cephalothin may suggest that the resistance patterns for farm environment samples may be more representative of the risk of contamination of surface waters with antimicrobial agent-resistant bacteria. PMID:15746342

Water-Related Infrastructure in a Region of Post-Earthquake Haiti: High Levels of Fecal Contamination and Need for Ongoing Monitoring

PubMed Central

Widmer, Jocelyn M.; Weppelmann, Thomas A.; Alam, Meer T.; Morrissey, B. David; Redden, Edsel; Rashid, Mohammed H.; Diamond, Ulrica; Ali, Afsar; De Rochars, Madsen Beau; Blackburn, Jason K.; Johnson, Judith A.; Morris, J. Glenn

2014-01-01

We inventoried non-surface water sources in the Leogane and Gressier region of Haiti (approximately 270 km2) in 2012 and 2013 and screened water from 345 sites for fecal coliforms and Vibrio cholerae. An international organization/non-governmental organization responsible for construction could be identified for only 56% of water points evaluated. Sixteen percent of water points were non-functional at any given time; 37% had evidence of fecal contamination, with spatial clustering of contaminated sites. Among improved water sources (76% of sites), 24.6% had fecal coliforms versus 80.9% in unimproved sources. Fecal contamination levels increased significantly from 36% to 51% immediately after the passage of Tropical Storm Sandy in October of 2012, with a return to 34% contamination in March of 2013. Long-term sustainability of potable water delivery at a regional scale requires ongoing assessment of water quality, functionality, and development of community-based management schemes supported by a national plan for the management of potable water. PMID:25071005

Water-related infrastructure in a region of post-earthquake Haiti: high levels of fecal contamination and need for ongoing monitoring.

PubMed

Widmer, Jocelyn M; Weppelmann, Thomas A; Alam, Meer T; Morrissey, B David; Redden, Edsel; Rashid, Mohammed H; Diamond, Ulrica; Ali, Afsar; De Rochars, Madsen Beau; Blackburn, Jason K; Johnson, Judith A; Morris, J Glenn

2014-10-01

We inventoried non-surface water sources in the Leogane and Gressier region of Haiti (approximately 270 km(2)) in 2012 and 2013 and screened water from 345 sites for fecal coliforms and Vibrio cholerae. An international organization/non-governmental organization responsible for construction could be identified for only 56% of water points evaluated. Sixteen percent of water points were non-functional at any given time; 37% had evidence of fecal contamination, with spatial clustering of contaminated sites. Among improved water sources (76% of sites), 24.6% had fecal coliforms versus 80.9% in unimproved sources. Fecal contamination levels increased significantly from 36% to 51% immediately after the passage of Tropical Storm Sandy in October of 2012, with a return to 34% contamination in March of 2013. Long-term sustainability of potable water delivery at a regional scale requires ongoing assessment of water quality, functionality, and development of community-based management schemes supported by a national plan for the management of potable water. © The American Society of Tropical Medicine and Hygiene.

Evaluation of Two PCR-based Swine-specific Fecal Source Tracking Assays (Abstract)

EPA Science Inventory

Several PCR-based methods have been proposed to identify swine fecal pollution in environmental waters. However, the utility of these assays in identifying swine fecal contamination on a broad geographic scale is largely unknown. In this study, we evaluated the specificity, distr...

Numbers of fecal streptococci and Escherichia coli in fresh and dry cattle, horse, and sheep manure.

PubMed

Weaver, R W; Entry, J A; Graves, Alexandria

2005-10-01

Livestock are known contributors to stream pollution. Numbers of fecal streptococci and Escherichia coli in manure naturally deposited by livestock in the field are needed for activities related to bacterial source tracking and determining maximum daily bacterial loading of streams. We measured populations of fecal streptococci and E. coli in fresh and dry manure from cattle (Bos taurus L.), horses (Equus caballus L.), and sheep (Ovis aires L.) on farms in southern Idaho. Populations of indicator bacteria in dry manure were often as high as that in fresh manure from horse and sheep. There was a 2 log10 drop in the population of fecal coliform numbers in dry cattle manure from cattle in pastures but not from cattle in pens. Bacterial isolates used in source tracking should include isolates from both fresh and dry manure to better represent the bacterial source loading of streams.

Detection of human fecal contamination by nifH gene quantification of marine waters in the coastal beaches of Rio de Janeiro, Brazil.

PubMed

Oliveira, Samara Sant'Anna; Sorgine, Marcos Henrique Ferreira; Bianco, Kayo; Pinto, Leonardo Henriques; Barreto, Camila; Albano, Rodolpho Mattos; Cardoso, Alexander Machado; Clementino, Maysa Mandetta

2016-12-01

The identification of fecal pollution in aquatic ecosystems is one of the requirements to assess the possible risks to human health. In this report, physicochemical parameters, Escherichia coli enumeration and Methanobrevibacter smithii nifH gene quantification were conducted at 13 marine waters in the coastal beaches of Rio de Janeiro, Brazil. The pH, turbidity, dissolved oxygen, temperature, and conductivity, carried out by mobile equipment, revealed varied levels due to specific conditions of the beaches. The bioindicators' enumerations were done by defined substrate method, conventional, and real-time PCR. Six marine beach sites (46 %) presenting E. coli levels in compliance with Brazilian water quality guidelines (<2500 MPN/100 mL) showed nifH gene between 5.7 × 10 9 to 9.5 × 10 11 copies. L -1 revealing poor correlation between the two approaches. To our knowledge, this is the first inquiry in qPCR using nifH gene as a biomarker of human-specific sources of sewage pollution in marine waters in Brazil. In addition, our data suggests that alternative indicator nifH gene could be used, in combination with other markers, for source tracking studies to measure the quality of marine ecosystems thereby contributing to improved microbial risk assessment.

A Strategy for Federal Science and Technology to Support Water Availability and Quality in the United States

DTIC Science & Technology

2007-09-01

Subcommittee on Water Availability and Quality Microbial source tracking to identify the source of fecal contamination Many of the Nation’s resource...water- quality health standards. In addition, identi- fication of fecal sources is relevant to source-water protection programs and to the development...estuaries, snowpack, and soil . • Develop sensors and systems to measure water quality inexpensively in real time. Develop innovative water-use technologies

Citizen-volunteer and professional monitoring to identify fecal sources of contamination in southwestern Puerto Rico

EPA Science Inventory

High concentrations of nutrients, fecal microorganisms, and sediments in surface waters can be a public health threat and can impact fringing coral reefs in Guánica Bay in southwestern Puerto Rico. Yet, the main factors and sources contributing to water quality degradation...

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...

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...

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...

Decay of Fecal Indicator Bacterial Populations and Bovine-Associated Source-Tracking Markers in Freshly Deposited Cow Pats

EPA Science Inventory

Understanding the survival of fecal indicator bacteria (FIB) and microbial source-tracking (MST) markers is critical to developing pathogen fate and transport models. Although pathogen survival in water microcosms and manure-amended soils is well documented, little is known about...

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...

DESIGN AND EVALUATION OF BACTEROIDES DNA PROBES FOR THE SPECIFIC DETECTION OF HUMAN FECAL POLLUTION

EPA Science Inventory

Because Bacteroides spp. are obligate anaerobes that dominate the human fecal flora, and because some species may live only in the human intestine, these bacteria might be useful to distinguish human from nonhuman sources of fecal pollution. To test this hypothesis, PCR primers s...

Correlative Assessment of Fecal Indicators Using Human Mitochondrial DNA as a Direct Marker

EPA Science Inventory

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 t...

Performance of human fecal anaerobe-associated PCR-based assays in a multi-laboratory method evaluation study

USGS Publications Warehouse

Layton, Blythe A.; Cao, Yiping; Ebentier, Darcy L.; Hanley, Kaitlyn; Ballesté, Elisenda; Brandão, João; Byappanahalli, Muruleedhara N.; Converse, Reagan; Farnleitner, Andreas H.; Gentry-Shields, Jennifer; Gourmelon, Michèle; Lee, Chang Soo; Lee, Jiyoung; Lozach, Solen; Madi, Tania; Meijer, Wim G.; Noble, Rachel; Peed, Lindsay; Reischer, Georg H.; Rodrigues, Raquel; Rose, Joan B.; Schriewer, Alexander; Sinigalliano, Chris; Srinivasan, Sangeetha; Stewart, Jill; ,; Laurie, C.; Wang, Dan; Whitman, Richard; Wuertz, Stefan; Jay, Jenny; Holden, Patricia A.; Boehm, Alexandria B.; Shanks, Orin; Griffith, John F.

2013-01-01

A number of PCR-based methods for detecting human fecal material in environmental waters have been developed over the past decade, but these methods have rarely received independent comparative testing in large multi-laboratory studies. Here, we evaluated ten of these methods (BacH, BacHum-UCD, Bacteroides thetaiotaomicron (BtH), BsteriF1, gyrB, HF183 endpoint, HF183 SYBR, HF183 Taqman®, HumM2, and Methanobrevibacter smithii nifH (Mnif)) using 64 blind samples prepared in one laboratory. The blind samples contained either one or two fecal sources from human, wastewater or non-human sources. The assay results were assessed for presence/absence of the human markers and also quantitatively while varying the following: 1) classification of samples that were detected but not quantifiable (DNQ) as positive or negative; 2) reference fecal sample concentration unit of measure (such as culturable indicator bacteria, wet mass, total DNA, etc); and 3) human fecal source type (stool, sewage or septage). Assay performance using presence/absence metrics was found to depend on the classification of DNQ samples. The assays that performed best quantitatively varied based on the fecal concentration unit of measure and laboratory protocol. All methods were consistently more sensitive to human stools compared to sewage or septage in both the presence/absence and quantitative analysis. Overall, HF183 Taqman® was found to be the most effective marker of human fecal contamination in this California-based study.

Trail Creek I: Assessing the Water Quality of Streams using Fecal Indicator Bacteria and Microbial Source Tracking

NASA Astrophysics Data System (ADS)

Saintil, T.; Radcliffe, D. E.; Rasmussen, T. C.; Kannan, A.

2017-12-01

Fecal coliforms are indicators for disease-causing pathogens. The United States Environmental Protection Agency (US. EPA) recommends the use of E. coli and Enterococci because they are highly correlated with pathogenic organisms in recreational waters. This standard method helps to determine the overall water quality and the potential health risks. Studies have shown that it is difficult to estimate the exact sources of fecal contamination because both human and certain animal species contain E. coli and Enterococci in their waste. Certain strains of E. coli and Enterococci are also able to survive outside of their hosts, which should not be the case for an appropriate fecal indicator. As a result, microbial source tracking (MST) studies use gene specific markers to identify the possible contributors to water pollution whether human or animal. Trail Creek is a second-order stream located in Athens-Clarke County, GA. The 33-km2 watershed is approximately 64% forests, 18% pastures and 16% residential communities. Trail Creek is on the TMDL list and an extended study on the relationships between the different factors causing elevated fecal bacteria is needed. Synoptic sampling events were conducted during baseflow conditions at six locations. Storm sampling events (> 8 mm) were captured using automated samplers at two locations. These samplers were equipped with pressure transducers which record stage at 30-minute intervals. The samples were analyzed for fecal coliform, E. Coli and Enterococci. Water quality parameters including temperature, specific conductance, dissolved oxygen, pH, and turbidity were also recorded. Relationships between the parameters and fecal indicator bacteria show inconsistent patterns and high variability. Using quantitative PCR and MST techniques, the human specific marker (HF183) and ruminant marker (Rum2Bac) were used to identify the fecal sources in both baseflow and storm samples. The presence and abundance of the different markers at each site will be presented. The findings will be useful for adopting best management practices and implementing green infrastructure for the reduction of fecal pollution.

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 detect sewage compared to general indicators, and the relationship to a human pathogen group suggests that the use of these alternative indicators will improve assessments for human health risks in urban waters. PMID:21803887

Interlaboratory comparison of three microbial source tracking quantitative polymerase chain reaction (qPCR) assays from fecal-source and environmental samples

USGS Publications Warehouse

Stelzer, Erin A.; Strickler, Kriston M.; Schill, William B.

2012-01-01

During summer and early fall 2010, 15 river samples and 6 fecal-source samples were collected in West Virginia. These samples were analyzed by three laboratories for three microbial source tracking (MST) markers: AllBac, a general fecal indicator; BacHum, a human-associated fecal indicator; and BoBac, a ruminant-associated fecal indicator. MST markers were analyzed by means of the quantitative polymerase chain reaction (qPCR) method. The aim was to assess interlaboratory precision when the three laboratories used the same MST marker and shared deoxyribonucleic acid (DNA) extracts of the samples, but different equipment, reagents, and analyst experience levels. The term assay refers to both the markers and the procedure differences listed above. Interlaboratory precision was best for all three MST assays when using the geometric mean absolute relative percent difference (ARPD) and Friedman's statistical test as a measure of interlaboratory precision. Adjustment factors (one for each MST assay) were calculated using results from fecal-source samples analyzed by all three laboratories and applied retrospectively to sample concentrations to account for differences in qPCR results among labs using different standards and procedures. Following the application of adjustment factors to qPCR results, ARPDs were lower; however, statistically significant differences between labs were still observed for the BacHum and BoBac assays. This was a small study and two of the MST assays had 52 percent of samples with concentrations at or below the limit of accurate quantification; hence, more testing could be done to determine if the adjustment factors would work better if the majority of sample concentrations were above the quantification limit.

Changes in Escherichia coli to Cryptosporidium ratios for various fecal pollution sources and drinking water intakes.

PubMed

Lalancette, Cindy; Papineau, Isabelle; Payment, Pierre; Dorner, Sarah; Servais, Pierre; Barbeau, Benoit; Di Giovanni, George D; Prévost, Michèle

2014-05-15

Assessing the presence of human pathogenic Cryptosporidium oocysts in surface water remains a significant water treatment and public health challenge. Most drinking water suppliers rely on fecal indicators, such as the well-established Escherichia coli (E. coli), to avoid costly Cryptosporidium assays. However, the use of E. coli has significant limitations in predicting the concentration, the removal and the transport of Cryptosporidium. This study presents a meta-analysis of E. coli to Cryptosporidium concentration paired ratios to compare their complex relationships in eight municipal wastewater sources, five agricultural fecal pollution sources and at 13 drinking water intakes (DWI) to a risk threshold based on US Environmental Protection Agency (USEPA) regulations. Ratios lower than the USEPA risk threshold suggested higher concentrations of oocysts in relation to E. coli concentrations, revealing an underestimed risk for Cryptosporidium based on E. coli measurements. In raw sewage (RS), high ratios proved E. coli (or fecal coliforms) concentrations were a conservative indicator of Cryptosporidium concentrations, which was also typically true for secondary treated wastewater (TWW). Removals of fecal indicator bacteria (FIB) and parasites were quantified in WWTPs and their differences are put forward as a plausible explanation of the sporadic ratio shift. Ratios measured from agricultural runoff surface water were typically lower than the USEPA risk threshold and within the range of risk misinterpretation. Indeed, heavy precipitation events in the agricultural watershed led to high oocyst concentrations but not to E. coli or enterococci concentrations. More importantly, ratios established in variously impacted DWI from 13 Canadian drinking water plants were found to be related to dominant fecal pollution sources, namely municipal sewage. In most cases, when DWIs were mainly influenced by municipal sewage, E. coli or fecal coliforms concentrations agreed with Cryptosporidium concentrations as estimated by the meta-analysis, but when DWIs were influenced by agricultural runoff or wildlife, there was a poor relationship. Average recovery values were available for 6 out of 22 Cryptosporidium concentration data sets and concomitant analysis demonstrated no changes in trends, with and without correction. Nevertheless, recovery assays performed along with every oocyst count would have enhanced the precision of this work. Based on our findings, the use of annual averages of E. coli concentrations as a surrogate for Cryptosporidium concentrations can result in an inaccurate estimate of the Cryptosporidium risk for agriculture impacted drinking water intakes or for intakes with more distant wastewater sources. Studies of upstream fecal pollution sources are recommended for drinking water suppliers to improve their interpretation of source water quality data. Copyright © 2014 Elsevier Ltd. All rights reserved.

Dynamics of Aeromonas hydrophila, Aeromonas sobria, and Aeromonas caviae in a sewage treatment pond.

PubMed Central

Monfort, P; Baleux, B

1990-01-01

The spatiotemporal dynamics of Aeromonas spp. and fecal coliforms in the sewage treatment ponds of an urban wastewater center were studied after 20 months of sampling from five stations in these ponds. Isolation and identification of 247 Aeromonas strains were undertaken over four seasons at the inflow and outflow of this pond system. The hemolytic activity of these strains was determined. The Aeromonas spp. and the fecal coliform distributions showed seasonal cycles, the amplitude of which increased at distances further from the wastewater source, so that in the last pond there was an inversion of the Aeromonas spp. cycle in comparison with that of fecal coliforms. The main patterns in these cycles occurred simultaneously at all stations, indicating control of these bacterial populations by seasonal factors (temperature, solar radiation, phytoplankton), the effects of which were different on each bacterial group. The analysis of the Aeromonas spp. population structure showed that, regardless of the season, Aeromonas caviae was the dominant species at the pond system inflow. However at the outflow the Aeromonas spp. population was dominated by A. caviae in winter, whereas Aeromonas sobria was the dominant species in the treated effluent from spring to fall. Among the Aeromonas hydrophila and A. sobria strains, 100% produced hemolysin; whereas among the A. caviae strains, 96% were nonhemolytic. Images PMID:2389929

Effect of dietary starch source and concentration on equine fecal microbiota

USDA-ARS?s Scientific Manuscript database

Starch from corn is less susceptible to equine small intestinal digestion than starch from oats, and starch that reaches the hindgut can be utilized by the microbiota. The objective of the current study was to examine the effects of starch source on equine fecal microbiota. Thirty horses were assig...

Effect of starch source in pelleted concentrates on fecal bacterial communities in Thoroughbred mares

USDA-ARS?s Scientific Manuscript database

High starch concentrates are often added to equine diets to meet digestible energy requirements of some horses, such as broodmares. Starch source has been shown to affect fecal bacterial communities of horses when fed cereal grains with little to no processing. Others suggest that grain processing, ...

Tracking the Primary Sources of Fecal Pollution in a Tropical Watershed in a One-Year Study

EPA Science Inventory

A study was conducted to determine the primary sources of fecal pollution in a subtropical watershed using host-specific assays developed in temperate regions. Water samples (n=534) from 10 different sites along the Rio Grande de Arecibo watershed were collected every two-three w...

Cost-Effectiveness Analysis of Six Strategies to Treat Recurrent Clostridium difficile Infection

PubMed Central

Lapointe-Shaw, Lauren; Tran, Kim L.; Coyte, Peter C.; Hancock-Howard, Rebecca L.; Powis, Jeff; Poutanen, Susan M.; Hota, Susy

2016-01-01

Objective To assess the cost-effectiveness of six treatment strategies for patients diagnosed with recurrent Clostridium difficile infection (CDI) in Canada: 1. oral metronidazole; 2. oral vancomycin; 3.oral fidaxomicin; 4. fecal transplantation by enema; 5. fecal transplantation by nasogastric tube; and 6. fecal transplantation by colonoscopy. Perspective Public insurer for all hospital and physician services. Setting Ontario, Canada. Methods A decision analytic model was used to model costs and lifetime health effects of each strategy for a typical patient experiencing up to three recurrences, over 18 weeks. Recurrence data and utilities were obtained from published sources. Cost data was obtained from published sources and hospitals in Toronto, Canada. The willingness-to-pay threshold was $50,000/QALY gained. Results Fecal transplantation by colonoscopy dominated all other strategies in the base case, as it was less costly and more effective than all alternatives. After accounting for uncertainty in all model parameters, there was an 87% probability that fecal transplantation by colonoscopy was the most beneficial strategy. If colonoscopy was not available, fecal transplantation by enema was cost-effective at $1,708 per QALY gained, compared to metronidazole. In addition, fecal transplantation by enema was the preferred strategy if the probability of recurrence following this strategy was below 8.7%. If fecal transplantation by any means was unavailable, fidaxomicin was cost-effective at an additional cost of $25,968 per QALY gained, compared to metronidazole. Conclusion Fecal transplantation by colonoscopy (or enema, if colonoscopy is unavailable) is cost-effective for treating recurrent CDI in Canada. Where fecal transplantation is not available, fidaxomicin is also cost-effective. PMID:26901316

Cost-Effectiveness Analysis of Six Strategies to Treat Recurrent Clostridium difficile Infection.

PubMed

Lapointe-Shaw, Lauren; Tran, Kim L; Coyte, Peter C; Hancock-Howard, Rebecca L; Powis, Jeff; Poutanen, Susan M; Hota, Susy

2016-01-01

To assess the cost-effectiveness of six treatment strategies for patients diagnosed with recurrent Clostridium difficile infection (CDI) in Canada: 1. oral metronidazole; 2. oral vancomycin; 3.oral fidaxomicin; 4. fecal transplantation by enema; 5. fecal transplantation by nasogastric tube; and 6. fecal transplantation by colonoscopy. Public insurer for all hospital and physician services. Ontario, Canada. A decision analytic model was used to model costs and lifetime health effects of each strategy for a typical patient experiencing up to three recurrences, over 18 weeks. Recurrence data and utilities were obtained from published sources. Cost data was obtained from published sources and hospitals in Toronto, Canada. The willingness-to-pay threshold was $50,000/QALY gained. Fecal transplantation by colonoscopy dominated all other strategies in the base case, as it was less costly and more effective than all alternatives. After accounting for uncertainty in all model parameters, there was an 87% probability that fecal transplantation by colonoscopy was the most beneficial strategy. If colonoscopy was not available, fecal transplantation by enema was cost-effective at $1,708 per QALY gained, compared to metronidazole. In addition, fecal transplantation by enema was the preferred strategy if the probability of recurrence following this strategy was below 8.7%. If fecal transplantation by any means was unavailable, fidaxomicin was cost-effective at an additional cost of $25,968 per QALY gained, compared to metronidazole. Fecal transplantation by colonoscopy (or enema, if colonoscopy is unavailable) is cost-effective for treating recurrent CDI in Canada. Where fecal transplantation is not available, fidaxomicin is also cost-effective.

IS THE IMMUNOCROMATOGRAPHIC FECAL ANTIGEN TEST EFFECTIVE FOR PRIMARY DIAGNOSIS OF HELICOBACTER PYLORI INFECTION IN DYSPEPTIC PATIENTS?

PubMed

Dalla Nora, Magali; Hörner, Rosmari; De Carli, Diego Michelon; Rocha, Marta Pires da; Araujo, Amanda Faria de; Fagundes, Renato Borges

2016-01-01

The diagnosis of H. pylori infection can be performed by non-invasive and invasive methods.The identification through a fecal antigen test is a non-invasive, simple, and relatively inexpensive test. To determine the diagnostic performance of fecal antigen test in the identification of H. pylori infection. H. pylori antigens were identified in the stools of dyspeptic patients undergoing upper gastrointestinal endoscopy. For the identification of H. pylori antigen, we use ImmunoCard STAT! HpSA with immunochromatography technique. Histopathology plus urease test were the gold standard. We studied 163 patients, 51% male, mean age of 56.7± 8.5years. H. pylori infection was present in 49%. Fecal test presented: sensitivity 67.5% (CI95% 60.6-72.9); specificity 85.5% (CI95% 78.9-90.7); positive predictive value 81.8% (CI95% 73.4-88.4) and negative predictive value 73,2% (CI95% 67.5-77.6); Positive likelihood ratio was 4.7 (CI95% 2.9-7.9) and Negative Likelihood Ratio 0.4 (CI95% 0.3-0.5). The prevalence odds ratio for a positive test was 12.3 (CI95% 5.7-26.3).The index kappa between FAT and histology/urease test was 0.53 (CI95% 0.39-0.64). Immunochromatographic FAT is less expensive than the other methods and readily accepted by the patients but its diagnostic performance does not recommend its use in the primary diagnosis, when the patient may have an active infection.

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

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.

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 detected at one site (Moose Brook) but was not detected at other sites. Fecal contamination (as indicated by fecal coliform density) apparently is localized under normal flow conditions with the highest levels restricted to drains in urban areas and to a lesser extent B Stream, Pearce Brook, and Big Brook, all tributaries to the main stem of the Meduxnekeag River. Coliphage were enumerated as an alternate indicator of fecal contamination with the intent of typing the virus into host-associated classes (human or ruminant), as was done for Enterococci; however, insufficient coliphage were isolated to provide more than preliminary indications. In spite of low coliphage enumeration, the preliminary results strengthen the conclusion that the Enterococci data correctly indicated the samples that contained human and ruminant fecal contamination. The finding that contamination was in many of the tributaries following storms in mid-July indicates that storm runoff likely carries fecal contaminants to more locations than runoff under lower flow conditions.

Animal Feces Contribute to Domestic Fecal Contamination: Evidence from E. coli Measured in Water, Hands, Food, Flies, and Soil in Bangladesh.

PubMed

Ercumen, Ayse; Pickering, Amy J; Kwong, Laura H; Arnold, Benjamin F; Parvez, Sarker Masud; Alam, Mahfuja; Sen, Debashis; Islam, Sharmin; Kullmann, Craig; Chase, Claire; Ahmed, Rokeya; Unicomb, Leanne; Luby, Stephen P; Colford, John M

2017-08-01

Fecal-oral pathogens are transmitted through complex, environmentally mediated pathways. Sanitation interventions that isolate human feces from the environment may reduce transmission but have shown limited impact on environmental contamination. We conducted a study in rural Bangladesh to (1) quantify domestic fecal contamination in settings with high on-site sanitation coverage; (2) determine how domestic animals affect fecal contamination; and (3) assess how each environmental pathway affects others. We collected water, hand rinse, food, soil, and fly samples from 608 households. We analyzed samples with IDEXX Quantitray for the most probable number (MPN) of E. coli. We detected E. coli in source water (25%), stored water (77%), child hands (43%), food (58%), flies (50%), ponds (97%), and soil (95%). Soil had >120 000 mean MPN E. coli per gram. In compounds with vs without animals, E. coli was higher by 0.54 log 10 in soil, 0.40 log 10 in stored water and 0.61 log 10 in food (p < 0.05). E. coli in stored water and food increased with increasing E. coli in soil, ponds, source water and hands. We provide empirical evidence of fecal transmission in the domestic environment despite on-site sanitation. Animal feces contribute to fecal contamination, and fecal indicator bacteria do not strictly indicate human fecal contamination when animals are present.

Animal Feces Contribute to Domestic Fecal Contamination: Evidence from E. coli Measured in Water, Hands, Food, Flies, and Soil in Bangladesh

PubMed Central

2017-01-01

Fecal-oral pathogens are transmitted through complex, environmentally mediated pathways. Sanitation interventions that isolate human feces from the environment may reduce transmission but have shown limited impact on environmental contamination. We conducted a study in rural Bangladesh to (1) quantify domestic fecal contamination in settings with high on-site sanitation coverage; (2) determine how domestic animals affect fecal contamination; and (3) assess how each environmental pathway affects others. We collected water, hand rinse, food, soil, and fly samples from 608 households. We analyzed samples with IDEXX Quantitray for the most probable number (MPN) of E. coli. We detected E. coli in source water (25%), stored water (77%), child hands (43%), food (58%), flies (50%), ponds (97%), and soil (95%). Soil had >120 000 mean MPN E. coli per gram. In compounds with vs without animals, E. coli was higher by 0.54 log10 in soil, 0.40 log10 in stored water and 0.61 log10 in food (p < 0.05). E. coli in stored water and food increased with increasing E. coli in soil, ponds, source water and hands. We provide empirical evidence of fecal transmission in the domestic environment despite on-site sanitation. Animal feces contribute to fecal contamination, and fecal indicator bacteria do not strictly indicate human fecal contamination when animals are present. PMID:28686435

Comparison of Microbial and Chemical Source Tracking Markers To Identify Fecal Contamination Sources in the Humber River (Toronto, Ontario, Canada) and Associated Storm Water Outfalls.

PubMed

Staley, Zachery R; Grabuski, Josey; Sverko, Ed; Edge, Thomas A

2016-11-01

Storm water runoff is a major source of pollution, and understanding the components of storm water discharge is essential to remediation efforts and proper assessment of risks to human and ecosystem health. In this study, culturable Escherichia coli and ampicillin-resistant E. coli levels were quantified and microbial source tracking (MST) markers (including markers for general Bacteroidales spp., human, ruminant/cow, gull, and dog) were detected in storm water outfalls and sites along the Humber River in Toronto, Ontario, Canada, and enumerated via endpoint PCR and quantitative PCR (qPCR). Additionally, chemical source tracking (CST) markers specific for human wastewater (caffeine, carbamazepine, codeine, cotinine, acetaminophen, and acesulfame) were quantified. Human and gull fecal sources were detected at all sites, although concentrations of the human fecal marker were higher, particularly in outfalls (mean outfall concentrations of 4.22 log 10 copies, expressed as copy numbers [CN]/100 milliliters for human and 0.46 log 10 CN/100 milliliters for gull). Higher concentrations of caffeine, acetaminophen, acesulfame, E. coli, and the human fecal marker were indicative of greater raw sewage contamination at several sites (maximum concentrations of 34,800 ng/liter, 5,120 ng/liter, 9,720 ng/liter, 5.26 log 10 CFU/100 ml, and 7.65 log 10 CN/100 ml, respectively). These results indicate pervasive sewage contamination at storm water outfalls and throughout the Humber River, with multiple lines of evidence identifying Black Creek and two storm water outfalls with prominent sewage cross-connection problems requiring remediation. Limited data are available on specific sources of pollution in storm water, though our results indicate the value of using both MST and CST methodologies to more reliably assess sewage contamination in impacted watersheds. Storm water runoff is one of the most prominent non-point sources of biological and chemical contaminants which can potentially degrade water quality and pose risks to human and ecosystem health. Therefore, identifying fecal contamination in storm water runoff and outfalls is essential for remediation efforts to reduce risks to public health. This study employed multiple methods of identifying levels and sources of fecal contamination in both river and storm water outfall sites, evaluating the efficacy of using culture-based enumeration of E. coli, molecular methods of determining the source(s) of contamination, and CST markers as indicators of fecal contamination. The results identified pervasive human sewage contamination in storm water outfalls and throughout an urban watershed and highlight the utility of using both MST and CST to identify raw sewage contamination. © Crown copyright 2016.

Comparison of Microbial and Chemical Source Tracking Markers To Identify Fecal Contamination Sources in the Humber River (Toronto, Ontario, Canada) and Associated Storm Water Outfalls

PubMed Central

Grabuski, Josey; Sverko, Ed; Edge, Thomas A.

2016-01-01

ABSTRACT Storm water runoff is a major source of pollution, and understanding the components of storm water discharge is essential to remediation efforts and proper assessment of risks to human and ecosystem health. In this study, culturable Escherichia coli and ampicillin-resistant E. coli levels were quantified and microbial source tracking (MST) markers (including markers for general Bacteroidales spp., human, ruminant/cow, gull, and dog) were detected in storm water outfalls and sites along the Humber River in Toronto, Ontario, Canada, and enumerated via endpoint PCR and quantitative PCR (qPCR). Additionally, chemical source tracking (CST) markers specific for human wastewater (caffeine, carbamazepine, codeine, cotinine, acetaminophen, and acesulfame) were quantified. Human and gull fecal sources were detected at all sites, although concentrations of the human fecal marker were higher, particularly in outfalls (mean outfall concentrations of 4.22 log10 copies, expressed as copy numbers [CN]/100 milliliters for human and 0.46 log10 CN/100 milliliters for gull). Higher concentrations of caffeine, acetaminophen, acesulfame, E. coli, and the human fecal marker were indicative of greater raw sewage contamination at several sites (maximum concentrations of 34,800 ng/liter, 5,120 ng/liter, 9,720 ng/liter, 5.26 log10 CFU/100 ml, and 7.65 log10 CN/100 ml, respectively). These results indicate pervasive sewage contamination at storm water outfalls and throughout the Humber River, with multiple lines of evidence identifying Black Creek and two storm water outfalls with prominent sewage cross-connection problems requiring remediation. Limited data are available on specific sources of pollution in storm water, though our results indicate the value of using both MST and CST methodologies to more reliably assess sewage contamination in impacted watersheds. IMPORTANCE Storm water runoff is one of the most prominent non-point sources of biological and chemical contaminants which can potentially degrade water quality and pose risks to human and ecosystem health. Therefore, identifying fecal contamination in storm water runoff and outfalls is essential for remediation efforts to reduce risks to public health. This study employed multiple methods of identifying levels and sources of fecal contamination in both river and storm water outfall sites, evaluating the efficacy of using culture-based enumeration of E. coli, molecular methods of determining the source(s) of contamination, and CST markers as indicators of fecal contamination. The results identified pervasive human sewage contamination in storm water outfalls and throughout an urban watershed and highlight the utility of using both MST and CST to identify raw sewage contamination. PMID:27542934

Presumptive Sources of Fecal Contamination in Four Tributaries to the New River Gorge National River, West Virginia, 2004

USGS Publications Warehouse

Mathes, Melvin V.; O'Brien, Tara L.; Strickler, Kriston M.; Hardy, Joshua J.; Schill, William B.; Lukasik, Jerzy; Scott, Troy M.; Bailey, David E.; Fenger, Terry L.

2007-01-01

Several methods were used to determine the sources of fecal contamination in water samples collected during September and October 2004 from four tributaries to the New River Gorge National River -- Arbuckle Creek, Dunloup Creek, Keeney Creek, and Wolf Creek. All four tributaries historically have had elevated levels of fecal coliform bacteria. The source-tracking methods used yielded various results, possibly because one or more methods failed. Sourcing methods used in this study included the detection of several human-specific and animal-specific biological or molecular markers, and library-dependent pulsed-field gel electrophoresis analysis that attempted to associate Escherichia coli bacteria obtained from water samples with animal sources by matching DNA-fragment banding patterns. Evaluation of the results of quality-control analysis indicated that pulsed-field gel electrophoresis analysis was unable to identify known-source bacteria isolates. Increasing the size of the known-source library did not improve the results for quality-control samples. A number of emerging methods, using markers in Enterococcus, human urine, Bacteroidetes, and host mitochondrial DNA, demonstrated some potential in associating fecal contamination with human or animal sources in a limited analysis of quality-control samples. All four of the human-specific markers were detected in water samples from Keeney Creek, a watershed with no centralized municipal wastewater-treatment facilities, thus indicating human sources of fecal contamination. The human-specific Bacteroidetes and host mitochondrial DNA markers were detected in water samples from Dunloup Creek, Wolf Creek, and to a lesser degree Arbuckle Creek. Results of analysis for wastewater compounds indicate that the September 27 sample from Arbuckle Creek contained numerous human tracer compounds likely from sewage. Dog, horse, chicken, and pig host mitochondrial DNA were detected in some of the water samples with the exception of the October 5 sample from Dunloup Creek. Cow, white-tailed deer, and Canada goose DNA were not detected in any of the samples collected from the four tributaries, despite the presence of these animals in the watersheds. Future studies with more rigorous quality-control analyses are needed to investigate the potential applicability and use of these emerging methods. Because many of the detections for the various methods could vary over time and with flow conditions, repeated sampling during both base flow and storm events would be necessary to more definitively determine the sources of fecal contamination for each watershed.

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...

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...

Effect of starch source (corn, oats or wheat) and concentration on fermentation by equine fecal microbiota in vitro

USDA-ARS?s Scientific Manuscript database

Aims: The goal was to determine the effect of starch source (corn, oats and wheat) and concentration on: 1) total amylolytic bacteria, Group D Gram-positive cocci (GPC), lactobacilli, and lactate-utilizing bacteria, and 2) fermentation by equine microflora. Methods and Results: When fecal washed cel...

EVALUATION OF SELECTED DNA-BASED TECHNOLOGY IN IMPAIRED WATERSHEDS IMPACTED BY FECAL CONTAMINATION FROM DIVERSE SOURCES

EPA Science Inventory

Fecal pollution of surface waters is a top reason for impairment, as listed by the U.S. Environmental Protection Agency’s report on the quality of the Nation's waters. To be able to develop and implement TMDLs in impaired aquatic resources, it is imperative to determine the sourc...

Evaluation of Water Quality for Two St. Johns River Tributaries Receiving Septic Tank Effluent, Duval County, Florida

USGS Publications Warehouse

Wicklein, Shaun M.

2004-01-01

Tributary streamflow to the St. Johns River in Duval County is thought to be affected by septic tank leachate from residential areas adjacent to these tributaries. Water managers and the city of Jacksonville have committed to infrastructure improvements as part of a management plan to address the impairment of tributary water quality. In order to provide data to evaluate the effects of future remedial activities in selected tributaries, major ion and nutrient concentrations, fecal coliform concentrations, detection of wastewater compounds, and tracking of bacterial sources were used to document septic tank influences on the water quality of selected tributaries. The tributaries Fishing Creek and South Big Fishweir Creek were selected because they drain subdivisions identified as high priority locations for septic tank phase-out projects: the Pernecia and Murray Hill B subdivisions, respectively. Population, housing (number of residences), and septic tank densities for the Murray Hill B subdivision are greater than those for the Pernecia subdivision. Water-quality samples collected in the study basins indicate influences from ground water and septic tanks. Estimated concentrations of total nitrogen ranged from 0.33 to 2.86 milligrams per liter (mg/L), and ranged from less than laboratory reporting limit (0.02 mg/L) to 0.64 mg/L for total phosphorus. Major ion concentrations met the State of Florida Class III surface-water standards; total nitrogen and total phosphorus concentrations exceeded the U.S. Environmental Protection Agency Ecoregion XII nutrient criteria for rivers and streams 49 and 96 percent of the time, respectively. Organic wastewater compounds detected at study sites were categorized as detergents, antioxidants and flame retardants, manufactured polycarbonate resins, industrial solvents, and mosquito repellent. The most commonly detected compound was para-nonylphenol, a breakdown product of detergent. Results of wastewater sampling give evidence that stream water in the study basins is affected by septic tank effluent. Fecal coliform bacteria concentrations were measured on a monthly basis; of 115 samples, 63 percent exceeded the State of Florida fecal coliform bacteria standard for Class III surface waters of 800 colonies per 100 milliliters of water on any 1 day. Fecal coliform bacteria concentrations ranged from less than 20 colonies per 100 milliliters of sample to greater than or equal to 160,000 colonies per 100 milliliters of sample. Antibiotic resistance patterns of fecal coliform bacteria were used to identify the sources of fecal coliform bacteria. Significant sources of fecal coliform bacteria included wild animals, dogs, and humans. A majority of the fecal coliform bacteria were classified to be from human sources. Because the primary source of fecal coliform bacteria is from human sources, and most likely septic tank effluent, management of human sources may substantially improve microbiological water quality in both the Fishing Creek and South Branch Big Fishweir Creek basins.

Fecal bile acids of black-footed ferrets

USGS Publications Warehouse

Richardson, Louise; Johnson, M.K.; Clark, T.W.; Schroder, M.H.

1986-01-01

Fecal bile acid characteristics have been used to identify scats to species of origin. Fecal bile acids in scats from 20 known black-footed ferrets ( Mustela nigripes ), 7 other known small carnivores, and 72 of unknown origin were analyzed to determine if this procedure could be used as a tool to verify ferret presence in an area. Seventeen ferret scats were suitable for analysis and had a mean fecal bile acid index of 156 ± 9. This was significantly different from mean indices for the other carnivores; however, substantial overlap among confidence intervals occurred for badgers, kit foxes, and especially long-tailed weasels. We conclude this method is not useful for making positive identifications if individual ferret scats and suggest that we may be able to definitively identify individual scats with reasonable confidence by using gas-liquid chromatography.

Use of Bacteroidales Microbial Source Tracking To Monitor Fecal Contamination in Fresh Produce Production

PubMed Central

Ravaliya, Kruti; 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. PMID:24212583

Human-Associated Bacteroides spp. and Human Polyomaviruses as Microbial Source Tracking Markers in Hawaii.

PubMed

Kirs, Marek; Caffaro-Filho, Roberto A; Wong, Mayee; Harwood, Valerie J; Moravcik, Philip; Fujioka, Roger S

2016-11-15

Identification of sources of fecal contaminants is needed to (i) determine the health risk associated with recreational water use and (ii) implement appropriate management practices to mitigate this risk and protect the environment. This study evaluated human-associated Bacteroides spp. (HF183TaqMan) and human polyomavirus (HPyV) markers for host sensitivity and specificity using human and animal fecal samples collected in Hawaii. The decay rates of those markers and indicator bacteria were identified in marine and freshwater microcosms exposed and not exposed to sunlight, followed by field testing of the usability of the molecular markers. Both markers were strongly associated with sewage, although the cross-reactivity of the HF183TaqMan (also present in 82% of canine [n = 11], 30% of mongoose [n = 10], and 10% of feline [n = 10] samples) needs to be considered. Concentrations of HF183TaqMan in human fecal samples exceeded those in cross-reactive animals at least 1,000-fold. In the absence of sunlight, the decay rates of both markers were comparable to the die-off rates of enterococci in experimental freshwater and marine water microcosms. However, in sunlight, the decay rates of both markers were significantly lower than the decay rate of enterococci. While both markers have their individual limitations in terms of sensitivity and specificity, these limitations can be mitigated by using both markers simultaneously; ergo, this study supports the concurrent use of HF183TaqMan and HPyV markers for the detection of sewage contamination in coastal and inland waters in Hawaii. This study represents an in-depth characterization of microbial source tracking (MST) markers in Hawaii. The distribution and concentrations of HF183TaqMan and HPyV markers in human and animal fecal samples and in wastewater, coupled with decay data obtained from sunlight-exposed and unexposed microcosms, support the concurrent application of HF183TaqMan and HPyV markers for sewage contamination detection in Hawaii waters. Both markers are more conservative and more specific markers of sewage than fecal indicator bacteria (enterococci and Escherichia coli). Analysis of HF183TaqMan (or newer derivatives) is recommended for inclusion in future epidemiological studies concerned with beach water quality, while better concentration techniques are needed for HPyV. Such epidemiological studies can be used to develop new recreational water quality criteria, which will provide direct information on the absence or presence of sewage contamination in water samples as well as reliable measurements of the risk of waterborne disease transmission to swimmers. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

Estuarine and Riverine Areas Final Programmatic Environmental Assessment

DTIC Science & Technology

2004-06-25

sources in the study area include WWTP spray field runoff, urban and agricultural runoff, septic tank leachate , landfill leachate , silviculture...overland sheet flow. Urban and agricultural runoff are sources of fecal and total coliform and fecal streptococcus bacteria. Septic tank leachate and...in leachate from experiments using sand showed the greatest mobility of tungsten. Outdoor exposures and accelerated aging tests studied the

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 five sampling sites along the 5.7-mi study reach of Shoal Creek, but the trends at successive downstream sites were out of phase and could not be explained by simple advection and dispersion. At base-flow conditions, the travel time of bacteria in Shoal Creek along the 5.7-mi reach between State Highway W (site 2) and the MDNR sampling site (site 3) was about 26 hours. Substantial dispersion and dilution occurs along the upper 4.1 mi of this reach because of inflows from a number of springs and tributaries and the presence of several long pools and channel meanders. Minimal dispersion and dilution occurs along the 1.6-mi reach immediately upstream from the MDNR sampling site. Measurements of fecal bacteria decay in Shoal Creek during July 2001 indicated that about 8 percent of fecal coliform and E. coli bacteria decay each hour with an average first-order decay constant of 0.084 h-1 (per hour). Results of field test plots indicated that substantial numbers of fecal bacteria present in poul try litter can survive in fields for as much as 8 weeks after the application of the litter to the land surface. Median densities of fecal coliform and E. coli in slurry-water samples collected from fields increased from less than 60 col/100 mL before the application of turkey and broiler litter, to as large as 420,000 and 290,000 col/100 mL after the application of litter. Bacteria densities in the test plots generally decreased in a exponential manner over time with decay rates ranging from 0.085 to 0.185 d-1 (per day) for fecal coliform to between 0.100 and 0.250 d-1 for E. coli. The apparent survival of significant numbers of fecal bacteria on fields where poultry litter has been applied indicates that runoff from these fields is a potential source of fecal bacteria to vicinity streams for many weeks following litter application.

Fecal contamination of drinking water within peri-urban households, Lima, Peru.

PubMed

Oswald, William E; Lescano, Andrés G; Bern, Caryn; Calderon, Maritza M; Cabrera, Lilia; Gilman, Robert H

2007-10-01

We assessed fecal contamination of drinking water in households in 2 peri-urban communities of Lima, Peru. We measured Escherichia coli counts in municipal source water and, within households, water from principal storage containers, stored boiled drinking water, and water in a serving cup. Source water was microbiologically clean, but 26 (28%) of 93 samples of water stored for cooking had fecal contamination. Twenty-seven (30%) of 91 stored boiled drinking water samples grew E. coli. Boiled water was more frequently contaminated when served in a drinking cup than when stored (P < 0.01). Post-source contamination increased successively through the steps of usage from source water to the point of consumption. Boiling failed to ensure safe drinking water at the point of consumption because of easily contaminated containers and poor domestic hygiene. Hygiene education, better point-of-use treatment and storage options, and in-house water connections are urgently needed.

Factors affecting isolation and identification of Mycobacterium avium subsp. paratuberculosis from fecal and tissue samples in a liquid culture system.

PubMed

Whittington, Richard J

2009-03-01

Culture of Mycobacterium avium subsp. paratuberculosis is the definitive diagnostic test for Johne's disease, a chronic granulomatous enteropathy of animals. Compared to solid media, the identification of all strains of the organism in liquid media can be more difficult because the appearance of colonies and mycobactin dependence are not observable, and the growth of other organisms needs to be distinguished, commonly by PCR. Factors affecting the isolation rate of S strains and the contamination rate in modified Middlebrook 7H9 broth (Bactec 12B) and 7H10 agar were studied using 11,598 fecal samples and 2,577 tissue samples from sheep from 1,421 farms over 10 years. Minimization of contamination in Bactec cultures required the avoidance of the carryover of fecal particles from the first sedimentation step in the double-incubation centrifugation method, and contamination was reduced significantly by incubating the sample in a solution containing vancomycin, amphotericin B, and nalidixic acid for 3 days compared to 2 days. The growth of irrelevant microorganisms confounded the identification of M. avium subsp. paratuberculosis in liquid culture by inhibiting IS900 PCR and in solid medium culture by inhibiting the growth of M. avium subsp. paratuberculosis or obscuring colonies. The contamination of samples was clustered in certain laboratory submissions and was reduced by including ampicillin in Bactec medium without affecting the odds of isolation of M. avium subsp. paratuberculosis. The long-term contamination rate for fecal cultures was about 7%, and that for tissue cultures was <0.2%. Liquid medium was more sensitive than solid medium culture for M. avium subsp. paratuberculosis. The applicability of these findings for C strains is discussed.

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 (Spearman's Rho value of 0.976). The majority of the fecal coliforms were Escherichia coli during the synoptic study. Fecal-indicator-bacteria concentrations were not correlated to streamflow, water temperature, dissolved oxygen, pH, specific conduc-tance, and alkalinity. Fecal-indicator-bacteria concentrations were moderately correlated with turbidity (Spearman's Rho values of 0.662 and 0.640 for fecal coliform and Escherichia coli, respectively) and sediment (Spearman's Rho values of 0.628 and 0.636 for fecal coliform and Escherichia coli, respectively). Escherichia coli isolates analyzed by discriminant analysis of ribotype patterns for samples from the Bighorn River at Basin, Wyoming, and Bitter Creek near Garland, Wyoming, in the Bighorn River Basin were determined to be from nonhuman and human sources. Using a confidence interval of 90 percent, more of the isolates from both sites were classified as being from nonhuman than human sources; however, both samples had additional isolates that were classified as unknown sources. --------------------------------------------------------------------------------

A study of fecal coliform sources at a coastal site using colored dissolved organic matter (CDOM) as a water source tracer.

PubMed

Clark, Catherine D; O'Connor, Adam P; Foley, Denise M; de Bruyn, Warren J

2007-09-01

Optical properties of colored dissolved organic matter (CDOM) were measured as a tracer of polluted waters in a Southern California surf-zone with consistently high levels of fecal indicator bacteria. Salinity, temperature, fecal coliform, absorbance (200-700nm) and fluorescence (lambda(excitation)=350nm; lambda(emission)=360-650nm) were measured in the creek and surf-zone during a dry and rain event. Fluorescence to absorption ratios for CDOM were used to distinguish water masses, with two distinct CDOM end-members identified as creek (flu/abs=8.7+/-0.8x10(4)) and coastal (flu/abs=2.2+/-0.3x10(4)). Waters containing the same CDOM end-member had highly variable bacterial levels during the dry event, suggesting intermittent sources of bacteria added to a uniform water source, consistent with marine birds. During the rain event, increased levels of the creek end-member and bacteria indicated a second bacteria source from runoff.

Microbial source tracking: a tool for identifying sources of microbial contamination in the food chain.

PubMed

Fu, Ling-Lin; Li, Jian-Rong

2014-01-01

The ability to trace fecal indicators and food-borne pathogens to the point of origin has major ramifications for food industry, food regulatory agencies, and public health. Such information would enable food producers and processors to better understand sources of contamination and thereby take corrective actions to prevent transmission. Microbial source tracking (MST), which currently is largely focused on determining sources of fecal contamination in waterways, is also providing the scientific community tools for tracking both fecal bacteria and food-borne pathogens contamination in the food chain. Approaches to MST are commonly classified as library-dependent methods (LDMs) or library-independent methods (LIMs). These tools will have widespread applications, including the use for regulatory compliance, pollution remediation, and risk assessment. These tools will reduce the incidence of illness associated with food and water. Our aim in this review is to highlight the use of molecular MST methods in application to understanding the source and transmission of food-borne pathogens. Moreover, the future directions of MST research are also discussed.

Beach boundary layer: a framework for addressing recreational water quality impairment at enclosed beaches.

PubMed

Grant, Stanley B; Sanders, Brett F

2010-12-01

Nearshore waters in bays, harbors, and estuaries are frequently contaminated with human pathogens and fecal indicator bacteria. Tracking down and mitigating this contamination is complicated by the many point and nonpoint sources of fecal pollution that can degrade water quality along the shore. From a survey of the published literature, we propose a conceptual and mathematical framework, the "beach boundary layer model", for understanding and quantifying the relative impact of beach-side and bay-side sources of fecal pollution on nearshore water quality. In the model, bacterial concentration in ankle depth water C(ankle) [bacteria L(-3)] depends on the flux m'' [bacteria L(-2) T(-1)] of fecal bacteria from beach-side sources (bather shedding, bird and dog feces, tidal washing of sediments, decaying vegetation, runoff from small drains, and shallow groundwater discharge), a cross-shore mass transfer velocity k [L T(-1)] that accounts for the physics of nearshore transport and mixing, and a background concentration C(bay) [bacteria L(-3)] attributable to bay-side sources of pollution that impact water quality over large regions (sewage outfalls, creeks and rivers): C(ankle) = m''/k + C(bay). We demonstrate the utility of the model for identifying risk factors and pollution sources likely to impact shoreline water quality, and evaluate the model's underlying assumptions using computational fluid dynamic simulations of flow, turbulence, and mass transport in a trapezoidal channel.

Assessment of sources of human pathogens and fecal contamination in a Florida freshwater lake.

PubMed

Staley, Christopher; Reckhow, Kenneth H; Lukasik, Jerzy; Harwood, Valerie J

2012-11-01

We investigated the potential for a variety of environmental reservoirs to harbor or contribute fecal indicator bacteria (FIB), DNA markers of human fecal contamination, and human pathogens to a freshwater lake. We hypothesized that submerged aquatic vegetation (SAV), sediments, and stormwater act as reservoirs and/or provide inputs of FIB and human pathogens to this inland water. Analysis included microbial source tracking (MST) markers of sewage contamination (Enterococcus faecium esp gene, human-associated Bacteroides HF183, and human polyomaviruses), pathogens (Salmonella, Cryptosporidium, Giardia, and enteric viruses), and FIB (fecal coliforms, Escherichia coli, and enterococci). Bayesian analysis was used to assess relationships among microbial and physicochemical variables. FIB in the water were correlated with concentrations in SAV and sediment. Furthermore, the correlation of antecedent rainfall and major rain events with FIB concentrations and detection of human markers and pathogens points toward multiple reservoirs for microbial contaminants in this system. Although pathogens and human-source markers were detected in 55% and 21% of samples, respectively, markers rarely coincided with pathogen detection. Bayesian analysis revealed that low concentrations (<45 CFU × 100 ml(-1)) of fecal coliforms were associated with 93% probability that pathogens would not be detected; furthermore the Bayes net model showed associations between elevated temperature and rainfall with fecal coliform and enterococci concentrations, but not E. coli. These data indicate that many under-studied matrices (e.g. SAV, sediment, stormwater) are important reservoirs for FIB and potentially human pathogens and demonstrate the usefulness of Bayes net analysis for water quality assessment. Copyright © 2012 Elsevier Ltd. All rights reserved.

OVERVIEW OF USEPA MICROBIOLOGICAL RESEARCH IN DRINKING WATER

EPA Science Inventory

The Microbial Contaminants Control Branch (MCCB) conducts research on microbiological problems related to drinking water treatment, distribution and storage, and has recently become involved in watershed and source water quality issues such as fecal indicator bacteria and fecal p...

Sudan stain of fecal fat: new insight into an old test.

PubMed

Khouri, M R; Huang, G; Shiau, Y F

1989-02-01

The 72-h fecal fat determination is used as the gold standard to document the presence of steatorrhea. Although the Sudan stain for fecal fat is advocated as a sensitive screening test, a quantitative correlation between the 72-h fecal fat quantitation and the fecal Sudan stain is lacking. This study was designed to examine the staining properties of different classes of purified lipids in an experimentally defined artificial matrix, and to elucidate the reasons for the lack of quantitative correlation between these two tests. Our results indicate that the "neutral fat" stain without acidification or heating identifies triglyceride; and at an appropriate pH, the "neutral stain" also identifies fatty acid. The "split fat" stain with acidification and heating identifies both triglyceride and fatty acid. After acidification, fatty acid soaps are converted to the nonionized fatty acid. Thus, fatty acid soaps can be identified indirectly as fat droplets that are stained by the split fat stain. Although cholesterol is stained with Sudan stain after heating, upon cooling, cholesterol forms crystals of anhydrous cholesterol, making its staining pattern distinct. Neither the neutral fat nor the split fat stain can detect phospholipid or cholesteryl ester. The 72-h fecal fat determination is a measure of the total fatty acid content after a specimen is saponified. The resulting fatty acids are derived from a variety of endogenous and exogenous sources, including free fatty acids, soaps of fatty acids, triglycerides, cholesterol esters, and phospholipids. Therefore, the 72-h fecal fat quantitation does not differentiate between the primary sources of the measured fatty acid. It is concluded that the 72-h fecal fat determination is not specific for documenting triglyceride (fat) malabsorption. Until new methods are developed that specifically measure fecal triglyceride and fatty acid, the Sudan stain of fecal fat appears to be a more specific method for detecting the presence of triglyceride and fatty acid in a matrix.

New Approaches to Attenuated Hepatitis a Vaccine Development: Cloning and Sequencing of Cell-Culture Adapted Viral cDNA

DTIC Science & Technology

1989-04-01

strain-specific identification of HAV in human fecal samples was a major aim of the original contract application, as clinical trials of live and...derived materials and human and primate fecal specimens. 4. We molecularly cloned and partially sequenced the genome of PA21 strain HAV, a virus...antibody. This approach revealed that 99% of the infectious virus particles present in disrupted cell lysates from the 23rd passage of persistently

Urban wet-weather flows: sources of fecal contamination impacting on recreational waters and threatening drinking-water sources.

PubMed

Marsalek, Jiri; Rochfort, Quintin

Discharges of urban stormwater and combined sewer overflows (CSOs) contribute to fecal contamination of urban waters and need to be considered in planning the protection of recreational waters and sources of drinking water. Stormwater characterization indicates that Escherichia coli counts in stormwater typically range from 103 to 104 units per 100 ml. Higher counts (10(5) units/100 ml) suggest the presence of cross-connections with sanitary sewers, and such connections should be identified and corrected. Fecal contamination of stormwater may be attenuated prior to discharge into surface waters by stormwater management measures, which typically remove suspended solids and attached bacteria. Exceptionally, stormwater discharges in the vicinity of swimming beaches are disinfected. The levels of indicator bacteria in CSOs can be as high as 10(6) E. coli per 100 ml. Consequently, the abatement of fecal contamination of CSOs is now considered in the design of CSO control and treatment, as for example stipulated in the Ontario Procedure F-5-5. CSO abatement options comprise combin ations of storage and treatment, in which the CSO treatment generally includes disinfection by ultraviolet (UV) irradiation. Finally, indicator bacteria data from Sarnia (Ontario) were used to demonstrate some fecal contamination impacts of wet-weather flows. In wet weather, the microbiological quality of riverine water worsened as a result of CSO and stormwater discharges, and the recreational water guidelines for indicator organisms were exceeded most of the time. Local improvements in water quality were feasible by source controls and diversion of polluted water.

Quantitative PCR measurements of Escherichia coli including shiga toxin-producing E. coli (STEC) in animal feces and environmental waters.

PubMed

Ahmed, W; Gyawali, P; Toze, S

2015-03-03

Quantitative PCR (qPCR) assays were used to determine the concentrations of E. coli including shiga toxin-producing E. coli (STEC) associated virulence genes (eaeA, stx1, stx2, and hlyA) in ten animal species (fecal sources) and environmental water samples in Southeast Queensland, Australia. The mean Log10 concentrations and standard deviations of E. coli 23S rRNA across fecal sources ranged from 1.3 ± 0.1 (horse) to 6.3 ± 0.4 (cattle wastewater) gene copies at a test concentration of 10 ng of DNA. The differences in mean concentrations of E. coli 23S rRNA gene copies among fecal source samples were significantly different from each other (P < 0.0001). Among the virulence genes, stx2 (25%, 95% CI, 17-33%) was most prevalent among fecal sources, followed by eaeA (19%, 95% CI, 12-27%), stx1 (11%, 95% CI, 5%-17%) and hlyA (8%, 95% CI, 3-13%). The Log10 concentrations of STEC virulence genes in cattle wastewater samples ranged from 3.8 to 5.0 gene copies at a test concentration of 10 ng of DNA. Of the 18 environmental water samples tested, three (17%) were positive for eaeA and two (11%) samples were also positive for the stx2 virulence genes. The data presented in this study will aid in the estimation of quantitative microbial risk assessment (QMRA) from fecal pollution of domestic and wild animals in drinking/recreational water catchments.

Multiple approaches to microbial source tracking in tropical northern Australia

PubMed Central

Neave, Matthew; Luter, Heidi; Padovan, Anna; Townsend, Simon; Schobben, Xavier; Gibb, Karen

2014-01-01

Microbial source tracking is an area of research in which multiple approaches are used to identify the sources of elevated bacterial concentrations in recreational lakes and beaches. At our study location in Darwin, northern Australia, water quality in the harbor is generally good, however dry-season beach closures due to elevated Escherichia coli and enterococci counts are a cause for concern. The sources of these high bacteria counts are currently unknown. To address this, we sampled sewage outfalls, other potential inputs, such as urban rivers and drains, and surrounding beaches, and used genetic fingerprints from E. coli and enterococci communities, fecal markers and 454 pyrosequencing to track contamination sources. A sewage effluent outfall (Larrakeyah discharge) was a source of bacteria, including fecal bacteria that impacted nearby beaches. Two other treated effluent discharges did not appear to influence sites other than those directly adjacent. Several beaches contained fecal indicator bacteria that likely originated from urban rivers and creeks within the catchment. Generally, connectivity between the sites was observed within distinct geographical locations and it appeared that most of the bacterial contamination on Darwin beaches was confined to local sources. PMID:25224738

Validation and application of quantitative PCR assays using host-specific Bacteroidales genetic markers for swine fecal pollution tracking.

PubMed

Fan, Lihua; Shuai, Jiangbing; Zeng, Ruoxue; Mo, Hongfei; Wang, Suhua; Zhang, Xiaofeng; He, Yongqiang

2017-12-01

Genome fragment enrichment (GFE) method was applied to identify host-specific bacterial genetic markers that differ among different fecal metagenomes. To enrich for swine-specific DNA fragments, swine fecal DNA composite (n = 34) was challenged against a DNA composite consisting of cow, human, goat, sheep, chicken, duck and goose fecal DNA extracts (n = 83). Bioinformatic analyses of 384 non-redundant swine enriched metagenomic sequences indicated a preponderance of Bacteroidales-like regions predicted to encode metabolism-associated, cellular processes and information storage and processing. After challenged against fecal DNA extracted from different animal sources, four sequences from the clone libraries targeting two Bacteroidales- (genes 1-38 and 3-53), a Clostridia- (gene 2-109) as well as a Bacilli-like sequence (gene 2-95), respectively, showed high specificity to swine feces based on PCR analysis. Host-specificity and host-sensitivity analysis confirmed that oligonucleotide primers and probes capable of annealing to select Bacteroidales-like sequences (1-38 and 3-53) exhibited high specificity (>90%) in quantitative PCR assays with 71 fecal DNAs from non-target animal sources. The two assays also demonstrated broad distributions of corresponding genetic markers (>94% positive) among 72 swine feces. After evaluation with environmental water samples from different areas, swine-targeted assays based on two Bacteroidales-like GFE sequences appear to be suitable quantitative tracing tools for swine fecal pollution. Copyright © 2017 Elsevier Ltd. All rights reserved.

40 CFR 432.25 - New source performance standards (NSPS).

Code of Federal Regulations, 2010 CFR

2010-07-01

..., fecal coliform, O&G, and TSS are the same as the limitations specified in § 432.22(a)(1), and the... on-site, the standards for BOD5, fecal coliform, O&G, and TSS are the same as the corresponding...

40 CFR 432.85 - New source performance standards (NSPS).

Code of Federal Regulations, 2011 CFR

2011-07-01

..., fecal coliform, O&G, and TSS specified in § 432.82(a). (b) Facilities that generate more than 50 million pounds per year of finished products must achieve the limitations for BOD5, fecal coliform, O&G, and TSS...

40 CFR 432.35 - New source performance standards (NSPS).

Code of Federal Regulations, 2011 CFR

2011-07-01

..., fecal coliform, TSS, and O&G are the same as the limitations specified in § 432.32(a)(1) and the... on-site, the standards for BOD5, fecal coliform, TSS, and O&G are the same as the corresponding...

40 CFR 432.65 - New source performance standards (NSPS).

Code of Federal Regulations, 2011 CFR

2011-07-01

..., fecal coliform, O&G, and TSS specified in § 432.62(a). (b) Facilities that generate more than 50 million pounds per year of finished products must achieve the limitations for BOD5, fecal coliform, O&G, and TSS...

40 CFR 432.35 - New source performance standards (NSPS).

Code of Federal Regulations, 2010 CFR

2010-07-01

..., fecal coliform, TSS, and O&G are the same as the limitations specified in § 432.32(a)(1) and the... on-site, the standards for BOD5, fecal coliform, TSS, and O&G are the same as the corresponding...

40 CFR 432.45 - New source performance standards (NSPS).

Code of Federal Regulations, 2010 CFR

2010-07-01

..., fecal coliform, TSS, and O&G are the same as the limitations specified in § 432.42(a)(1); and standards... on-site, the standards for BOD5, fecal coliform, TSS, and O&G are the same as the limitations...

40 CFR 432.15 - New source performance standards (NSPS).

Code of Federal Regulations, 2011 CFR

2011-07-01

..., fecal coliform, O&G, and TSS are the same as the corresponding limitations specified in § 432.12(a)(1... on-site, the standards for BOD5, fecal coliform, O&G, and TSS are the same as the limitations...

40 CFR 432.25 - New source performance standards (NSPS).

Code of Federal Regulations, 2011 CFR

2011-07-01

..., fecal coliform, O&G, and TSS are the same as the limitations specified in § 432.22(a)(1), and the... on-site, the standards for BOD5, fecal coliform, O&G, and TSS are the same as the corresponding...

40 CFR 432.85 - New source performance standards (NSPS).

Code of Federal Regulations, 2010 CFR

2010-07-01

..., fecal coliform, O&G, and TSS specified in § 432.82(a). (b) Facilities that generate more than 50 million pounds per year of finished products must achieve the limitations for BOD5, fecal coliform, O&G, and TSS...

40 CFR 432.15 - New source performance standards (NSPS).

Code of Federal Regulations, 2010 CFR

2010-07-01

..., fecal coliform, O&G, and TSS are the same as the corresponding limitations specified in § 432.12(a)(1... on-site, the standards for BOD5, fecal coliform, O&G, and TSS are the same as the limitations...

40 CFR 432.45 - New source performance standards (NSPS).

Code of Federal Regulations, 2011 CFR

2011-07-01

..., fecal coliform, TSS, and O&G are the same as the limitations specified in § 432.42(a)(1); and standards... on-site, the standards for BOD5, fecal coliform, TSS, and O&G are the same as the limitations...

Reductions in Gull Populations Improve Beachwater Quality.

EPA Science Inventory

BACKGROUND. Gulls are often cited as an important source of fecal pollution to surface waters, and fecal indicator bacteria (FIB) concentrations have been shown to be significantly correlated with gull populations. However, it is unclear whether gull contamination poses a risk to...

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.

Male-specific coliphages for source tracking fecal contamination in surface waters and prevalence of Shiga-toxigenic Escherichia coli in a major produce production region of the Central Coast of California.

PubMed

Ravva, Subbarao V; Sarreal, Chester Z; Cooley, Michael B

2015-07-01

To provide data for traditional trace-back studies from fork to farm, it is necessary to determine the environmental sources for Shiga-toxigenic Escherichia coli. We developed SYBR green based reverse-transcriptase PCR methods to determine the prevalence of F+ RNA coliphages (FRNA) as indicators of fecal contamination. Male-specific coliphages, determined using a single-agar overlay method, were prevalent in all surface waters sampled for 8 months. F+ DNA coliphages (FDNA) were predominant compared to FRNA in water samples from majority of sampling locations. Most (90%) of the FRNA were sourced to humans and originated from human-impacted sites. Members of genogroup III represented 77% of FRNA originated from human sources. Furthermore, 93% of FRNA sourced to animals were also detected in water samples from human-impacted sites. Eighty percent of all FRNA were isolated during the winter months indicating seasonality in prevalence. In contrast, FDNA were more prevalent during summer months. E. coli O157:H7 and Shiga-toxigenic E. coli were detected in water samples from locations predominantly influenced by agriculture. Owing to their scarcity, their numbers could not be correlated with the prevalence of FRNA or FDNA in water samples. Both coliform bacteria and generic E. coli from agricultural or human-impacted sites were similar in numbers and thus could not be used to determine the sources of fecal contamination. Data on the prevalence of male-specific coliphages may be invaluable for predicting the sources of fecal contamination and aid in developing methods to prevent enteric pathogen contamination from likely sources during produce production.

Application of SELECT and SWAT models to simulate source load, fate, and transport of fecal bacteria in watersheds.

NASA Astrophysics Data System (ADS)

Ranatunga, T.

2017-12-01

Modeling of fate and transport of fecal bacteria in a watershed is 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 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 from SELECT model were input to the SWAT model, and simulate the bacteria transport through the land and in-stream. The calibrated SWAT model was then used to estimate the indicator bacteria in-stream concentrations for future years based on regional land use, population and household forecast (up to 2040). Based on the reductions required to meet the water quality standards in-stream, the corresponding required source load reductions were estimated.

Evaluation of fecal contamination by human and ruminant sources in upper Fountain Creek, Colorado, 2007-2008, by using multiple lines of evidence:

USGS Publications Warehouse

Stoeckel, Donald

2011-01-01

Fountain Creek is a high-gradient stream on the Front Range of the Rocky Mountains in Colorado. The headwaters of Fountain Creek drain Pikes Peak, a major destination for tourism. Fountain Creek is a drinking-water source for the City of Colorado Springs, Colorado, and is used for irrigation, recreation, and other purposes between Colorado Springs and the confluence with the Arkansas River at Pueblo, Colorado. In 2008, Fountain Creek was placed on the Colorado 303(d) list of impaired streams because of fecal contamination. Colorado uses a 30-day geometric mean standard of 126 Escherichia coli per 100 milliliters as its management goal for recreational waters. The objective of this study was to identify major sources of Escherichia coli in upper Fountain Creek during exceedances of the State recreational water standard. To meet this objective, a new approach was developed and tested that uses genetic marker analysis for microbial source tracking, along with other information, to evaluate potential contributions of fecal contamination from various sources.

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 catchment scale by analysing three rivers impacted by fecal contaminations. The development and the application of this MST toolbox have highlighted (1) the specificity of the aforementioned markers, (2) their conservative transfer from soils to rivers and (3) their difference of persistence in seawater and in freshwater. Those results provide useful data in order to identify and manage fecal contaminations of superficial waters. In the case of single source contaminations, the markers provide coherent information: (1) the bovine or porcine markers were not detected in a river impacted by a WWTP effluent; (2) the occurrence of Rum-2-Bac and the distribution of stanols indicated a bovine contamination in a river flowing through cattle pasture. In the case of multiple source contaminations, the combination of markers is necessary to identify the main sources and the statistical treatment of the distribution of stanols could provide an approximation of their proportion.

Bacterial source tracking guides management of boat head waste in a coastal resort area.

PubMed

Mallin, Michael A; Haltom, Mary I; Song, Bongkeun; Tavares, Mary E; Dellies, Stephen P

2010-12-01

Fecal contamination of water bodies causes a public health problem and economic loss. To control such contamination management actions need to be guided by sound science. From 2007-2009 a study was undertaken to determine the sources of fecal bacteria contamination to the marine waters adjoining the Town of Wrightsville Beach, North Carolina, USA. The research effort included sampling for fecal coliform and Enterococcus bacteria, sampling for optical brighteners, dye studies, and use of molecular bacterial source tracking techniques including polymerase chain reaction (PCR) and terminal restriction fragment polymorphism (T-RFLP) fingerprinting of the Bacteroides-Prevotella group. Of the 96 samples collected from nine locations during the study, the water contact standard for Enterococcus was exceeded on 13 occasions. The T-RFLP fingerprint analyses demonstrated that the most widespread source of fecal contamination was human, occurring in 38% of the samples, with secondary ruminant and avian sources also detected. Optical brightener concentrations were low, reflecting a lack of sewage line leakage or spills. A lack of sewer leaks and lack of septic systems in the town pointed toward discharge from boat heads into the marine waters as the major cause of fecal contamination; this was supported by dye studies. Based on these data, the Town initiated action to have the U.S. Environmental Protection Agency declare the coastal waters (out to 3 nautical miles), the nearby Atlantic Intracoastal Waterway and its tributaries a no-discharge zone (NDZ) to alleviate the human fecal pollution. The Town garnered supporting resolutions from other local communities who jointly petitioned the North Carolina Department of Environmental and Natural Resources. This State regulatory agency supported the local government resolutions and sent an application for an NDZ to the EPA in April 2009. The EPA concurred, and in February 2010 the coastal waters of New Hanover County, NC, became the first marine area on the U.S. eastern seaboard between Delaware and the Florida Keys to be declared a no-discharge zone. Copyright © 2010 Elsevier Ltd. All rights reserved.

40 CFR 432.75 - New source performance standards (NSPS).

Code of Federal Regulations, 2011 CFR

2011-07-01

... achieve the standards for BOD5, fecal coliform, O&G, and TSS specified in § 432.72(a). (b) Facilities that..., fecal coliform, O&G, and TSS specified in § 432.72(b) and the limitations for ammonia (as N) and total...

Bifidobacteria in Feces and Environmental Waters

EPA Science Inventory

Bifidobacteria have been recommended as potential indicators of human fecal pollution in surface waters even though very little is known about their presence in nonhuman fecal sources. The objective of this research was to shed light on the occurrence and molecular diversity of t...

Targeting the Sources of Fecal Contamination using Dog-, Human-, and Ruminant- Specific Markers in the Lake Herrick Watershed, Georgia.

NASA Astrophysics Data System (ADS)

Saintil, T.; Radcliffe, D. E.; Rasmussen, T. C.; Habteselassie, M.; Sowah, R.; Kannan, A.

2016-12-01

The Lake Herrick Watershed is about 1.5 km2 and covers portions of the University of Georgia's East campus, the Oconee Forest, residential and commercial landuse. Lake Herrick, a recreational site on the University of Georgia campus, was closed in 2002 due to fecal contamination. Subsequent monitoring confirmed persistent contamination, which led to a permanent closure to swimming, boating, and fishing. While fecal coliform abundance is a standard metric for determining human health risks, Geldreich (1970) showed that fecal abundance does not necessarily correlate with the presence of pathogens. Nor does it identify pollution sources, which are needed to mitigate health risks. Two inflow tributaries and the outlet stream were monitored for discharge, fecal coliform, forms of nitrogen and phosphorus and other water-quality data to quantify lake influent and effluent bacteria loads. Fecal sources were identified using the human HF183 genetic marker (Seurinck et al., 2005), the ruminant BacR marker (Reischer et al., 2006), and the dog mitochondrial DNA (mtDNA) marker (Tambalo et al., 2012). Preliminary results confirm high concentrations of E. coli and Enterococci, above the State's limit of 124 MPN/100 mL, in both baseflows and stormflows. The findings also suggest that the E. coli and Enterococci loads from the inlet tributaries are on average higher compared to the bacteria loads coming out of the outlet stream. The human markers were detectable at all three sites but most of the samples were not quantifiable. The ruminant markers were quantifiable at both inlets but no ruminant markers were found at the outlet. The dog markers were detectable but not quantifiable at both inlets and no dog markers were detected at the outlet. Statistical analyses will be used to establish relationships between the nutrients data, the fecal concentrations, and the gene-specific markers.

Assessing hog lagoon waste contamination in the Cape Fear Watershed using Bacteroidetes 16S rRNA gene pyrosequencing.

PubMed

Arfken, Ann M; Song, Bongkeun; Mallin, Michael A

2015-09-01

Hog lagoons can be major sources of waste and nutrient contamination to watersheds adjacent to pig farms. Fecal source tracking methods targeting Bacteroidetes 16S rRNA genes in pig fecal matter may underestimate or fail to detect hog lagoon contamination in riverine environments. In order to detect hog lagoon wastewater contamination in the Cape Fear Watershed, where a large number of hog farms are present, we conducted pyrosequencing analyses of Bacteroidetes 16S rRNA genes in hog lagoon waste and identified new hog lagoon-specific marker sequences. Additional pyrosequencing analyses of Bacteroidetes 16S rRNA genes were conducted with surface water samples collected at 4 sites during 5 months in the Cape Fear Watershed. Using an operational taxonomic unit (OTU) identity cutoff value of 97 %, these newly identified hog lagoon markers were found in 3 of the river samples, while only 1 sample contained the pig fecal marker. In the sample containing the pig fecal marker, there was a relatively high percentage (14.1 %) of the hog lagoon markers and a low pig fecal marker relative abundance of 0.4 % in the Bacteroidetes 16S rRNA gene sequences. This suggests that hog lagoon contamination must be somewhat significant in order for pig fecal markers to be detected, and low levels of hog lagoon contamination cannot be detected targeting only pig-specific fecal markers. Thus, new hog lagoon markers have a better detection capacity for lagoon waste contamination, and in conjunction with a pig fecal marker, provide a more comprehensive and accurate detection of hog lagoon waste contamination in susceptible watersheds.

Storm drains are sources of human fecal pollution during dry weather in three urban southern California watersheds.

PubMed

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

2009-01-15

Coastal urbanized areas in Southern California experience frequent beach water quality warnings in summer due to high concentrations of fecal indicator bacteria (FIB). Remediation can be difficult, as sources are often unknown. During two summers, we sampled three urbanized watersheds in Santa Barbara, CA at sites with historically high FIB concentrations to determine if human fecal matter was influencing water quality. By quantification of a human-specific Bacteroides marker (HBM), human waste was evidenced throughout both transects, and concentrations were highest in the discharges of several flowing storm drains. The HBM concentrations in storm drain discharges varied by up to 5 orders of magnitude on the same day. While the exact points of entry into the storm drain systems were not definitively determined, further inspection of the drain infrastructure suggested exfiltrating sanitary sewers as possible sources. The HBM and FIB concentrations were not consistently correlated, although the exclusive occurrence of high HBM concentrations with high FIB concentrations warrants the use of FIB analyses for a first tier of sampling. The association of human fecal pollution with dry weather drainage could be a window into a larger problem for other urbanized coastal areas with Mediterranean-type climates.

Probabilistic quantitative microbial risk assessment model of norovirus from wastewater irrigated vegetables in Ghana using genome copies and fecal indicator ratio conversion for estimating exposure dose.

PubMed

Owusu-Ansah, Emmanuel de-Graft Johnson; Sampson, Angelina; Amponsah, Samuel K; Abaidoo, Robert C; Dalsgaard, Anders; Hald, Tine

2017-12-01

The need to replace the commonly applied fecal indicator conversions ratio (an assumption of 1:10 -5 virus to fecal indicator organism) in Quantitative Microbial Risk Assessment (QMRA) with models based on quantitative data on the virus of interest has gained prominence due to the different physical and environmental factors that might influence the reliability of using indicator organisms in microbial risk assessment. The challenges facing analytical studies on virus enumeration (genome copies or particles) have contributed to the already existing lack of data in QMRA modelling. This study attempts to fit a QMRA model to genome copies of norovirus data. The model estimates the risk of norovirus infection from the intake of vegetables irrigated with wastewater from different sources. The results were compared to the results of a corresponding model using the fecal indicator conversion ratio to estimate the norovirus count. In all scenarios of using different water sources, the application of the fecal indicator conversion ratio underestimated the norovirus disease burden, measured by the Disability Adjusted Life Years (DALYs), when compared to results using the genome copies norovirus data. In some cases the difference was >2 orders of magnitude. All scenarios using genome copies met the 10 -4 DALY per person per year for consumption of vegetables irrigated with wastewater, although these results are considered to be highly conservative risk estimates. The fecal indicator conversion ratio model of stream-water and drain-water sources of wastewater achieved the 10 -6 DALY per person per year threshold, which tends to indicate an underestimation of health risk when compared to using genome copies for estimating the dose. Copyright © 2017 Elsevier B.V. All rights reserved.

Occurrence of bacteriophages infecting Aeromonas, Enterobacter, and Klebsiella in water and association with contamination sources in Thailand.

PubMed

Wangkahad, Bencharong; Bosup, Suchada; Mongkolsuk, Skorn; Sirikanchana, Kwanrawee

2015-06-01

The co-residence of bacteriophages and their bacterial hosts in humans, animals, and environmental sources directed the use of bacteriophages to track the origins of the pathogenic bacteria that can be found in contaminated water. The objective of this study was to enumerate bacteriophages of Aeromonas caviae (AecaKS148), Enterobacter sp. (EnspKS513), and Klebsiella pneumoniae (KlpnKS648) in water and evaluate their association with contamination sources (human vs. animals). Bacterial host strains were isolated from untreated wastewater in Bangkok, Thailand. A double-layer agar technique was used to detect bacteriophages. All three bacteriophages were detected in polluted canal samples, with likely contamination from human wastewater, whereas none was found in non-polluted river samples. AecaKS148 was found to be associated with human fecal sources, while EnspKS513 and KlpnKS648 seemed to be equally prevalent in both human and animal fecal sources. Both bacteriophages were also present in polluted canals that could receive contamination from other fecal sources or the environment. In conclusion, all three bacteriophages were successfully monitored in Bangkok, Thailand. This study provided an example of bacteriophages for potential use as source identifiers of pathogen contamination. The results from this study will assist in controlling sources of pathogen contamination, especially in developing countries.

Characterizing the temporal patterns of avian influenza virus introduction into Japan by migratory birds

PubMed Central

ONUMA, Manabu; KAKOGAWA, Masayoshi; YANAGISAWA, Masae; HAGA, Atsushi; OKANO, Tomomi; NEAGARI, Yasuko; OKANO, Tsukasa; GOKA, Koichi; ASAKAWA, Mitsuhiko

2017-01-01

The objectives of the present study were to observe the temporal pattern of avian influenza virus (AIV) introduction into Japan and to determine which migratory birds play an important role in introducing AIV. In total, 19,407 fecal samples from migratory birds were collected at 52 sites between October 2008 and May 2015. Total nucleic acids extracted from the fecal samples were subjected to reverse transcription loop–mediated isothermal amplification to detect viral RNA. Species identification of host migratory birds was conducted by DNA barcoding for positive fecal samples. The total number of positive samples was 352 (prevalence, 1.8%). The highest prevalence was observed in autumn migration, and a decrease in prevalence was observed. During autumn migration, central to southern Japan showed a prevalence higher than the overall prevalence. Thus, the main AIV entry routes may involve crossing the Sea of Japan and entry through the Korean Peninsula. Species identification was successful in 221 of the 352 positive samples. Two major species sequences were identified: the Mallard/Eastern Spot-billed duck group (115 samples; 52.0%) and the Northern pintail (61 samples; 27.6%). To gain a better understanding of the ecology of AIV in Japan and the introduction pattern of highly pathogenic avian influenza viruses, information regarding AIV prevalence by species, the prevalence of hatch-year migratory birds, migration patterns and viral subtypes in fecal samples using egg inoculation and molecular-based methods in combination is required. PMID:28484128

Validation of a fecal scoring scale in puppies during the weaning period.

PubMed

Grellet, Aurélien; Feugier, Alexandre; Chastant-Maillard, Sylvie; Carrez, Bruno; Boucraut-Baralon, Corine; Casseleux, Gregory; Grandjean, Dominique

2012-10-01

In puppies weaning is a high risk period. Fecal changes are frequent and can be signs of infection by digestive pathogens (bacteria, viruses, parasites) and indicators of nutritional and environmental stress. The aim of this study was to define a pathological fecal score for weaning puppies, and to study the impact on that score of two intestinal viruses (canine parvovirus type 2 and canine coronavirus). For this, the quality of stools was evaluated on 154 puppies between 4 and 8 weeks of age (100 from small breeds and 54 from large breeds). The scoring was performed immediately after a spontaneous defecation based on a 13-point scale (from 1; liquid to 13; dry and hard feces). Fecal samples were frozen for further viral analysis. Each puppy was weighed once a week during the study period. The fecal score regarded as pathological was the highest score associated with a significant reduction in average daily gain (ADG). Fecal samples were checked by semi-quantitative PCR or RT-PCR for canine parvovirus type 2 and canine coronavirus identification, respectively. The quality of feces was affected by both age and breed size. In small breeds, the ADG was significantly reduced under a fecal score of 6 and 7 for puppies at 4-5 and 6-8 weeks of age, respectively. In large breeds, the ADG was significantly reduced under a fecal score of 5 whatever the age of the puppy. Whereas a high viral load of canine parvovirus type 2 significantly impacted feces quality, no effect was recorded for canine coronavirus. This study provides an objective threshold for evaluation of fecal quality in weaning puppies. It also emphasizes the importance to be given to age and breed size in that evaluation. Copyright © 2012 Elsevier B.V. All rights reserved.

Association of Supply Type with Fecal Contamination of Source Water and Household Stored Drinking Water in Developing Countries: A Bivariate Meta-analysis

PubMed Central

Bain, Robert E.S.; Cronk, Ryan; Wright, Jim A.; Bartram, Jamie

2015-01-01

Background Access to safe drinking water is essential for health. Monitoring access to drinking water focuses on water supply type at the source, but there is limited evidence on whether quality differences at the source persist in water stored in the household. Objectives We assessed the extent of fecal contamination at the source and in household stored water (HSW) and explored the relationship between contamination at each sampling point and water supply type. Methods We performed a bivariate random-effects meta-analysis of 45 studies, identified through a systematic review, that reported either the proportion of samples free of fecal indicator bacteria and/or individual sample bacteria counts for source and HSW, disaggregated by supply type. Results Water quality deteriorated substantially between source and stored water. The mean percentage of contaminated samples (noncompliance) at the source was 46% (95% CI: 33, 60%), whereas mean noncompliance in HSW was 75% (95% CI: 64, 84%). Water supply type was significantly associated with noncompliance at the source (p < 0.001) and in HSW (p = 0.03). Source water (OR = 0.2; 95% CI: 0.1, 0.5) and HSW (OR = 0.3; 95% CI: 0.2, 0.8) from piped supplies had significantly lower odds of contamination compared with non-piped water, potentially due to residual chlorine. Conclusions Piped water is less likely to be contaminated compared with other water supply types at both the source and in HSW. A focus on upgrading water services to piped supplies may help improve safety, including for those drinking stored water. Citation Shields KF, Bain RE, Cronk R, Wright JA, Bartram J. 2015. Association of supply type with fecal contamination of source water and household stored drinking water in developing countries: a bivariate meta-analysis. Environ Health Perspect 123:1222–1231; http://dx.doi.org/10.1289/ehp.1409002 PMID:25956006

Effects of Cereal, Fruit and Vegetable Fibers on Human Fecal Weight and Transit Time: A Comprehensive Review of Intervention Trials

PubMed Central

de Vries, Jan; Birkett, Anne; Hulshof, Toine; Verbeke, Kristin; Gibes, Kernon

2016-01-01

Cereal fibers are known to increase fecal weight and speed transit time, but far less data are available on the effects of fruits and vegetable fibers on regularity. This study provides a comprehensive review of the impact of these three fiber sources on regularity in healthy humans. We identified English-language intervention studies on dietary fibers and regularity and performed weighted linear regression analyses for fecal weight and transit time. Cereal and vegetable fiber groups had comparable effects on fecal weight; both contributed to it more than fruit fibers. Less fermentable fibers increased fecal weight to a greater degree than more fermentable fibers. Dietary fiber did not change transit time in those with an initial time of <48 h. In those with an initial transit time ≥48 h, transit time was reduced by approximately 30 min per gram of cereal, fruit or vegetable fibers, regardless of fermentability. Cereal fibers have been studied more than any other kind in relation to regularity. This is the first comprehensive review comparing the effects of the three major food sources of fiber on bowel function and regularity since 1993. PMID:26950143

Effects of Cereal, Fruit and Vegetable Fibers on Human Fecal Weight and Transit Time: A Comprehensive Review of Intervention Trials.

PubMed

de Vries, Jan; Birkett, Anne; Hulshof, Toine; Verbeke, Kristin; Gibes, Kernon

2016-03-02

Cereal fibers are known to increase fecal weight and speed transit time, but far less data are available on the effects of fruits and vegetable fibers on regularity. This study provides a comprehensive review of the impact of these three fiber sources on regularity in healthy humans. We identified English-language intervention studies on dietary fibers and regularity and performed weighted linear regression analyses for fecal weight and transit time. Cereal and vegetable fiber groups had comparable effects on fecal weight; both contributed to it more than fruit fibers. Less fermentable fibers increased fecal weight to a greater degree than more fermentable fibers. Dietary fiber did not change transit time in those with an initial time of <48 h. In those with an initial transit time ≥48 h, transit time was reduced by approximately 30 min per gram of cereal, fruit or vegetable fibers, regardless of fermentability. Cereal fibers have been studied more than any other kind in relation to regularity. This is the first comprehensive review comparing the effects of the three major food sources of fiber on bowel function and regularity since 1993.

Epidemiological survey and analysis on an outbreak of gastroenteritis due to water contamination.

PubMed

Yang, ZhiCong; Wu, XinWei; Li, TieGang; Li, MeiXia; Zhong, Yi; Liu, YuFei; Deng, Zhiai; DI, Biao; Huang, Cong; Liang, HuiYing; Wang, Ming

2011-06-01

To document the investigation and control of an outbreak of gastroenteritis in City G, South China, and provide a reference for preventing future outbreaks. An ambispective cohort study was designed. Attack rate (AR) and relative risks (RR) were calculated to identify the causes of gastroenteritis. Investigations using questionnaires included personal interviews with patients and doctors, reviews of medical records, laboratory examinations of fecal specimens and continuous hygiene monitoring of water samples from the waterworks. Overall, 427/71534 (AR=5.97%) cases were identified between October 31 and November 12 2010. Geographic distribution was highly localized, with 80% of cases occurring in the areas supplied by waterworks-A. Consumption of water provided solely by waterworks-A was found to be associated with illness (RR=8.20, 95 CI%:6.12-10.99) compared with that from waterworks-B. Microbiological analyses confirmed the presence of Norovirus in six of eight fecal samples from symptomatic patients, two water samples from waterworks-A and two sewage samples. After taking effective measures, the hygienic indices of waterworks-A met health criteria again on November 9 and no cases were reported 3 days later. The outbreak reported here was caused by drinking tap water contaminated with sewage at the source. Early identification of possible contamination sources and awareness of changes that might negatively impact water quality are important preventive measures to protect public health. Copyright © 2011 The Editorial Board of Biomedical and Environmental Sciences. Published by Elsevier B.V. All rights reserved.

Cheyletiella infestation in cats.

PubMed

Fox, J G; Hewes, K

1976-08-01

While routinely examining a fecal specimen of a cat for evidence of intestinal parasitism, large eggs (230 by 110 mu) were observed. tfurther examination of the fecal specimen revealed an adult Cheyletiella sp. mite. Of 42 cats received from the same source, 11 had Cheyletiella mites, found either on the skin or in fecal specimens. Grossly visible skin changes were not observed in any of the cats. The conically shaped sensory organ on genu I of the mite resembled that of Cheyletiella blakei.

Data for distribution of various species of fecal coliforms in urban, rural and private drinking water sources in ten years period - A case study: Kermanshah, Iran.

PubMed

Davoodi, Reza; Pirsaheb, Meghdad; Karimyan, Kamaladdin; Gupta, Vinod Kumar; Takhtshahi, Ali Reza; Sharafi, Hooshmand; Moradi, Masoud

2018-06-01

This study was aimed to investigate the distribution of various species of fecal coliform in urban, rural and private drinking water sources of Kermanshah, in the west of Iran. For this study, data of ten years period (2006-2016) assessments of microbial quality regarding various species of Fecal coliforms was taken from health centers associated with urban, rural and private resources of Kermanshah city. A total number of 8643 samples were taken, 1851 samples from rural, 365 from urban and 4834 from private resources. The results showed that Fecal coliforms , Escherichia coli ( E. coli ) had the widest distribution in all urban, rural and private water resources (22.3%, 45.9% and 34%, respectively). Moreover, E. coli (47.5%) and Klebsiella (0.4%) had, respectively, the highest and lowest distribution in all months considered. Based on the results, E.coli exists mostly in water resources; it is therefore of particular importance in the monitoring of water resources.

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.

Microbes in the neonatal intensive care unit resemble those found in the gut of premature infants

PubMed Central

2014-01-01

Background The source inoculum of gastrointestinal tract (GIT) microbes is largely influenced by delivery mode in full-term infants, but these influences may be decoupled in very low birth weight (VLBW, <1,500 g) neonates via conventional broad-spectrum antibiotic treatment. We hypothesize the built environment (BE), specifically room surfaces frequently touched by humans, is a predominant source of colonizing microbes in the gut of premature VLBW infants. Here, we present the first matched fecal-BE time series analysis of two preterm VLBW neonates housed in a neonatal intensive care unit (NICU) over the first month of life. Results Fresh fecal samples were collected every 3 days and metagenomes sequenced on an Illumina HiSeq2000 device. For each fecal sample, approximately 33 swabs were collected from each NICU room from 6 specified areas: sink, feeding and intubation tubing, hands of healthcare providers and parents, general surfaces, and nurse station electronics (keyboard, mouse, and cell phone). Swabs were processed using a recently developed ‘expectation maximization iterative reconstruction of genes from the environment’ (EMIRGE) amplicon pipeline in which full-length 16S rRNA amplicons were sheared and sequenced using an Illumina platform, and short reads reassembled into full-length genes. Over 24,000 full-length 16S rRNA sequences were produced, generating an average of approximately 12,000 operational taxonomic units (OTUs) (clustered at 97% nucleotide identity) per room-infant pair. Dominant gut taxa, including Staphylococcus epidermidis, Klebsiella pneumoniae, Bacteroides fragilis, and Escherichia coli, were widely distributed throughout the room environment with many gut colonizers detected in more than half of samples. Reconstructed genomes from infant gut colonizers revealed a suite of genes that confer resistance to antibiotics (for example, tetracycline, fluoroquinolone, and aminoglycoside) and sterilizing agents, which likely offer a competitive advantage in the NICU environment. Conclusions We have developed a high-throughput culture-independent approach that integrates room surveys based on full-length 16S rRNA gene sequences with metagenomic analysis of fecal samples collected from infants in the room. The approach enabled identification of discrete ICU reservoirs of microbes that also colonized the infant gut and provided evidence for the presence of certain organisms in the room prior to their detection in the gut. PMID:24468033

Evidence of Avian and Possum Fecal Contamination in Rainwater Tanks as Determined by Microbial Source Tracking Approaches

PubMed Central

Hamilton, K. A.; Gyawali, P.; Toze, S.; Haas, C. N.

2016-01-01

ABSTRACT Avian and possum fecal droppings may negatively impact roof-harvested rainwater (RHRW) water quality due to the presence of zoonotic pathogens. This study was aimed at evaluating the performance characteristics of a possum feces-associated (PSM) marker by screening 210 fecal and wastewater samples from possums (n = 20) and a range of nonpossum hosts (n = 190) in Southeast Queensland, Australia. The host sensitivity and specificity of the PSM marker were 0.90 and 0.95 (maximum value, 1.00), respectively. The mean concentrations of the GFD marker in possum fecal DNA samples (8.8 × 107 gene copies per g of feces) were two orders of magnitude higher than those in the nonpossum fecal DNA samples (5.0 × 105 gene copies per g of feces). The host sensitivity, specificity, and concentrations of the avian feces-associated GFD marker were reported in our recent study (W. Ahmed, V. J. Harwood, K. Nguyen, S. Young, K. Hamilton, and S. Toze, Water Res 88:613–622, 2016, http://dx.doi.org/10.1016/j.watres.2015.10.050). The utility of the GFD and PSM markers was evaluated by testing a large number of tank water samples (n = 134) from the Brisbane and Currumbin areas. GFD and PSM markers were detected in 39 of 134 (29%) and 11 of 134 (8%) tank water samples, respectively. The GFD marker concentrations in PCR-positive samples ranged from 3.7 × 102 to 8.5 × 105 gene copies per liter, whereas the concentrations of the PSM marker ranged from 2.0 × 103 to 6.8 × 103 gene copies per liter of water. The results of this study suggest the presence of fecal contamination in tank water samples from avian and possum hosts. This study has established an association between the degradation of microbial tank water quality and avian and possum feces. Based on the results, we recommend disinfection of tank water, especially for tanks designated for potable use. IMPORTANCE The use of roof-harvested rainwater (RHRW) for domestic purposes is a globally accepted practice. The presence of pathogens in rainwater tanks has been reported by several studies, supporting the necessity for the management of potential health risks. The sources of fecal pollution in rainwater tanks are unknown. However, the application of microbial source tracking (MST) markers has the potential to identify the sources of fecal contamination in a rainwater tank. In this study, we provide evidence of avian and possum fecal contamination in tank water samples using molecular markers. This study established a potential link between the degradation of the microbial quality of tank water and avian and possum feces. PMID:27208100

Point-of-use Unit Based on Gravity Ultrafiltration Removes Waterborne Gastrointestinal Pathogens from Untreated Water Sources in Rural Communities.

PubMed

Chaidez, Cristóbal; Ibarra-Rodríguez, Juan R; Valdez-Torres, José Benigno; Soto, Marcela; Gerba, Charles P; Castro-Del Campo, Nohelia

2016-09-01

In developing countries, rural communities often face the lack of potable water infrastructure and must rely on untreated sources for drinking, which are often contaminated with waterborne pathogens. The use of home water treatment devices is seen as one means of reducing the risk of exposure to waterborne pathogens. The aim of this study was to evaluate the microbiological and physicochemical performance of a simple in-home point-of-use device based on gravity ultrafiltration through an ultrafilter membrane. Twenty-five randomly selected households from 2 rural communities in Culiacán, Mexico, were enrolled. Water samples were collected before and after treatment and during storage for a period of 8 weeks. Heterotrophic bacteria, total coliforms, fecal coliforms, Escherichia coli, and Giardia spp were quantified, as well as various physicochemical parameters. All of the untreated water samples contained high levels of indicator bacteria, but none were detected in the treated water fulfilling the requirements set by the Mexican Norm (NOM-127-SSA1-1994) and the World Health Organization guidelines for drinking water. However, indicator bacteria (fecal coliforms and E coli) were detected in every sample from water stored 24 hours after treatment. This study demonstrated that point-of-use filters using gravity-fed ultrafilters are a low-cost, effective water treatment technology for water of poor microbial quality. However, further identification of the sources and mechanisms by which water is contaminated when stored after treatment will help with designing and implementing better strategies for keeping water safe for domestic use. Copyright © 2016 Wilderness Medical Society. Published by Elsevier Inc. All rights reserved.

Alteration of the fecal microbiota in Chinese patients with Parkinson's disease.

PubMed

Qian, Yiwei; Yang, Xiaodong; Xu, Shaoqing; Wu, Chunyan; Song, Yanyan; Qin, Nan; Chen, Sheng-Di; Xiao, Qin

2018-05-01

Emerging evidences suggest that gut microbiota dysbiosis plays a role in Parkinson's disease (PD). However, the alterations in fecal microbiome in Chinese PD patients remains unknown. This case-control study was conducted to explore fecal microbiota compositions in Chinese PD patients. Microbiota communities in the feces of 45 patients and their healthy spouses were investigated using high-throughput Illumina Miseq sequencing targeting the V3-V4 region of 16S ribosomal RNA (rRNA) gene. The relationships between fecal microbiota and PD clinical characteristics were analyzed. The structure and richness of the fecal microbiota differed between PD patients and healthy controls. Genera Clostridium IV, Aquabacterium, Holdemania, Sphingomonas, Clostridium XVIII, Butyricicoccus and Anaerotruncus were enriched in the feces of PD patients after adjusting for age, gender, body mass index (BMI), and constipation. Furthermore, genera Escherichia/Shigella were negatively associated with disease duration. Genera Dorea and Phascolarctobacterium were negatively associated with levodopa equivalent doses (LED). Among the non-motor symptoms (NMSs), genera Butyricicoccus and Clostridium XlVb were associated with cognitive impairment. Overall, we confirmed that gut microbiota dysbiosis occurs in Chinese patients with PD. A well-controlled population involved was beneficial for the identification of microbiota associated with diseases. Additionally, the fecal microbiota was closely related to PD clinical characteristics. Elucidating these differences in the fecal microbiome will provide a foundation to improve our understanding the pathogenesis of PD and to support the potentially therapeutic options modifying the gut microbiota. Copyright © 2018 Elsevier Inc. All rights reserved.

Molecular Characterization of Bacterial Populations in Sandhill Crane Feces

EPA Science Inventory

Sandhill cranes have the potential to be important seasonal sources of waterborne fecal bacteria; however, it is currently not possible to measure the extent of crane fecal pollution in potentially impacted areas as there are no available crane-specific assays. This is in part d...

EPA Technology Available for Licensing: Viral-Based Real-Time Quantitative PCR Test for Human Fecal Contamination

EPA Pesticide Factsheets

Human fecal contamination of clean water sources is a major contributor to the spread of disease worldwide. To monitor and manage this threat, the United States Environmental Protection Agency (EPA), World Health Organization, and European Union rely on ba

Current and future trends in fecal source tracking and deployment in the Lake Taihu Region of China

NASA Astrophysics Data System (ADS)

Hagedorn, Charles; Liang, Xinqiang

The emerging discipline of microbial and/or chemical source tracking (collectively termed fecal source tracking (FST)) is being used to identify origins of fecal contamination in polluted waters in many countries around the world. FST has developed rapidly because standard methods of measuring contamination in water by enumerating fecal indicator bacteria (FIB) such as fecal coliforms and enterococci do not identify the sources of the contamination. FST is an active area of research and development in both the academic and private sectors and includes: Developing and testing new microbial and chemical FST methods. Determining the geographic application and animal host ranges of existing and emerging FST techniques. Conducting experimental comparisons of FST techniques. Combining direct monitoring of human pathogens associated with waterborne outbreaks and zoonotic pathogens responsible for infections among people, wildlife, or domesticated animals with the use of FST techniques. Applying FST to watershed analysis and coastal environments. Designing appropriate statistical and probability analysis of FST data and developing models for mass loadings of host-specific fecal contamination. This paper includes a critical review of FST with emphasis on the extent to which methods have been tested (especially in comparison with other methods and/or with blind samples), which methods are applicable to different situations, their shortcomings, and their usefulness in predicting public health risk or pathogen occurrence. In addition, the paper addresses the broader question of whether FST and fecal indicator monitoring is the best approach to regulate water quality and protect human health. Many FST methods have only been tested against sewage or fecal samples or isolates in laboratory studies (proof of concept testing) and/or applied in field studies where the “real” answer is not known, so their comparative performance and accuracy cannot be assessed. For FST to be quantitative, stability of ratios between host-specific markers in the environment must be established. In addition, research is needed on the correlation between host-specific markers and pathogens, and survival of markers after waste treatments. As a result of the exclusive emphasis on FIB by regulatory agencies, monitoring and FST development has concentrated on FIB rather than the actual pathogens. A more rational approach to regulating water quality might be to use available epidemiological data to identify pathogens of concern in a particular water body, and then use targeted pathogen monitoring coupled with very specific FST approaches to control the pathogens. Baseline monitoring of FIB would be just one tool among many in this example.

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 water. Sediments are likely to be a relatively less important source of fecal bacteria during rainfall and runoff in the middle main stem relative to bacterial loading from point sources. Numerical streamflow and transport simulation models were calibrated and verified with data collected during field studies. Of the constituents modeled, bacteria exhibited the poorest correspondence between observed and simulated values. The simulation results for a dye tracer indicated that the model reasonably reproduced the timing of dissolved constituents as well as dilution and dispersion effects. Calibrated and verified models for 1991 and 1992 data sets were used to simulate the improvements to bacteriological water quality that might result from reductions in concentrations of fecal bacteria discharged from two major sources. The model simulation resulting in the greatest improvement in bacteriological water-quality was one in which concentrations of fecal coliform bacteria and Escherichia coli were reduced by 90 percent in the Cuyahoga River at the Old Portage gaging station, and to geometric-mean bathing-water standards in the effluent of the Akron Water Pollution Control Station (BWS/90 scenario). Compared to the results of the base-simulation, when the BWS/90 scenario was applied in the 1991 model simulation, Escherichia coli concentrations were reduced 98.5 percent at Botzum, 97.5 percent at Jaite, and 91.1 percent at Independence. For 1992 model simulations, similar percent reductions in the concentrations of Escherichia coli were predicted at the three stream sites when the same reductions were applied to sources. None of the model simulations resulted in attainment of bacteriological water-quality standards.The potential benefits of source reductions to human health and recreational uses were estimated by comparing the number of illnesses per 1,000 people from concentrations of Escherichia coli associated with the BWS/90 simulation, with the base simulation, and with the geometric-mean standard for Escherichia coli. The predicted 22 to 26 illnesses per 1,000 people predicted by the E. coli concentrations resulting from BWS/90 simulation are 2.8 to 3.3 times higher than the 8 illnesses per 1,000 people associated with the geometric-mean primary-contact water-quality standard for Escherichia coli. Risks associated with the base simulation are 4.6 to 4.9 times higher than that associated with the geometric-mean primary- contact water-quality standard for Escherichia coli. The illness risks predicted from the BWS/90 scenario, although larger than acceptable, would nevertheless be an improvement over conditions that were encountered during field studies in 1991-93.

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 Oakmont. For this site, dilution during wet-weather events or the lack of source streams upgradient of the site may have caused this anomaly. Additionally, single-event concentrations of E. coli and FC frequently exceeded the WQS reported during wet-weather events. It is difficult to establish a short-term trend in fecal-indicator bacteria concentrations as a function of time after a wet-weather event due to factors including the spatial variability of sources contributing fecal material, dry-weather discharges, resuspension of bottom sediments, and flow augmentation from reservoirs. Relative to E. coli and enterococci, FC concentrations appeared to decrease with time, which may be attributed to the greater die-off rate for FC bacteria. Fecal-indicator bacteria concentrations at a site are dependent on the spatial distribution of point sources upstream of the station, the time-of-travel, rate of decay, and the degree of mixing and resuspension. Therefore, it is difficult to evaluate whether the left, right, and composite concentrations reported at a particular site are significantly different. To evaluate the significance of the fecal-indicator bacteria concentrations and turbidity reported in grab and composite samples during dry-, mixed-, and wet-weather events, data sets were evaluated using Wilcoxon rank sum tests. Tests were conducted using the fecal-indicator bacteria colonies and turbidity reported for each station for a given weather event. For example, fecal coliform counts reported in the left-bank sample were compared against the right-bank and composite samples, respectively, for the Ohio River at Sewickley site during dry-, mixed-, and wet-weather events. The statistical analyses suggest that, depending on the sampling site, the fecal-bacteria concentrations measured at selected locations vary spatially within a channel (left bank compared to right, right bank compared to composite). The most significant differences occurred between feca

Assessment of the relative sensitivity of milk ELISA for detection of Mycobacterium avium ssp. paratuberculosis infectious dairy cows.

PubMed

Laurin, Emilie L; Sanchez, Javier; Chaffer, Marcelo; McKenna, Shawn L B; Keefe, Greg P

2017-01-01

Milk ELISA are commonly used for detection of Mycobacterium avium ssp. paratuberculosis (MAP) antibodies in dairy cows, due to low cost and quick processing for large numbers of samples. However, low sensitivity and variations from host and environmental factors can impede detection of MAP antibodies at early disease stages. The objectives of our study were to assess the sensitivity of milk ELISA in comparison with fecal tests and to evaluate how detectable antibody concentrations in milk vary with changes in fecal shedding of MAP, cow age, cow parity, days in milk, and time of year. To compare the sensitivity of a commercial milk ELISA with solid and broth fecal culture and with fecal real-time PCR, a longitudinal study was performed for the identification of MAP-infectious animals as determined by prior fecal testing for MAP shedding. In addition, associations between variation in milk MAP ELISA score and changes in fecal MAP shedding, host age, days in milk, and season were evaluated. Monthly milk and fecal samples were collected over 1 yr from 46 cows that were previously shedding MAP in their feces. Sensitivity of milk ELISA was 29.9% (95% CI: 24.8 to 35.1%), compared with 46.7% (40.7 to 52.7%) for fecal solid culture, 55.0% (49.3 to 60.7%) for fecal broth culture, and 78.4% (73.3 to 83.1%) for fecal direct real-time PCR. The effect of stage of lactation could not be separated from the effect of season, with increased milk ELISA scores at greater days in milk in winter. However, unpredictable monthly variations in results were observed among the 3 assays for individual cow testing, which highlights the importance of identifying patterns in pathogen and antibody detection over time in MAP-positive herds. Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

The Effects of Water Matrix on Decay of Human Fecal Molecular Markers and Campylobacter spp.

EPA Science Inventory

Although molecular source tracking for human fecal contamination is used on a wide range of sample types, little is known about comparative decay of proposed molecular markers under different conditions, or correlation with pathogen decay. Our purpose was to measure correlations ...

PERSISTENCE OF PHARMACEUTICALS AND OTHER WASTEWATER RELATED COMPOUNDS: UTILITY AS INDICATORS OF HUMAN FECAL CONTAMINATION

EPA Science Inventory

The quality of drinking and recreational water is currently ascertained using indicator bacteria. The tests to analyze for these bacteria require a considerable length of time to complete, and do not discriminate between human and animal fecal material sources. To shorten the t...

Distribution of Gull Specific Molecular Marker in Coastal Areas of Lake Ontario

EPA Science Inventory

Gulls have been implicated as primary sources of fecal contamination in the Great Lakes, a fact that may have health implications due to the potential spread of microbial pathogens by waterfowl. To better understand the spatial variability of gull fecal contamination, a gull-spe...

Can QMRA be used to Discount Pathogen Risk to Swimmers from Animal Fecal Contamination? Doheny Beach, CA Case Study

EPA Science Inventory

Estimated health risks to swimmers from seagull and bather sources of fecal contamination at Doheny Beach, California were compared using quantitative microbial risk assessment (QMRA) with a view to aiding beach closure decisions. Surfzone pathogens from seagulls were thought to...

An Assessment of Fecal Contamination in Rural Streams Impacted by Litter Applications

EPA Science Inventory

The objective of this study was to evaluate transport of fecal materials into streams located in farms containing pastures with the presence (AH and DH) and absence (DF) of cattle using various source-specific PCR assays. All pasture sites were fertilized with composted chicken l...

MOLECULAR TRACKING FECAL CONTAMINATION IN SURFACE WATERS: 16S RDNA VERSUS METAGENOMICS APPROACHES

EPA Science Inventory

Microbial source tracking methods need to be sensitive and exhibit temporal and geographic stability in order to provide meaningful data in field studies. The objective of this study was to use a combination of PCR-based methods to track cow fecal contamination in two watersheds....

INFLUENCE OF EQUINE FECAL CONTAMINATION ON MICROBIAL COMMUNITY STRUCTURE IN A SUB-WATERSHED

EPA Science Inventory

As monitoring of watershed water quality is directly related to proposed uses, it is important for our Naton's water systems that the d4etemrination of and distinction between fecal contamination source is made. The most common water quality monitoring approach is to screen for f...

From Sewers to Salix and Tailpipes to Typha: Riparian Plants Reflect Anthropogenic Nitrogen Sources Across Montane to Urban Gradients

NASA Astrophysics Data System (ADS)

Hall, S. J.; Hale, R. L.; Baker, M. A.; Bowling, D. R.; Ehleringer, J. R.

2014-12-01

Urban and suburban streams typically receive anthropogenic nitrogen (N) from multiple sources, and their identification and partitioning is a prerequisite for effective water quality management. However, stream N fluxes and sources are often highly variable, limiting the utility of water samples for source identification. Nitrate in perennial streams can provide an important N source for riparian vegetation in semi-arid environments. Thus, riparian plant tissue may integrate the stable isotope composition (δ15N) of stream nitrate over longer timescales and assist in source identification. Here, we tested whether δ15N of riparian plant leaves could provide an effective indicator of spatial variation in N sources across land use gradients spanning wildland to urban ecosystems in Salt Lake City, Utah, and the surrounding Wasatch Range Megapolitan Area. We found that leaf δ15N varied systematically within and among eight streams and rivers (n = 378 leaf samples) consistent with spatial land use variations. Plants from a suburban stream adjacent to homes with septic systems (δ15N = 5.1‰) were highly enriched relative to similar species from an adjacent undeveloped stream (δ15N = -0.7 ‰), suggesting an important contribution of enriched human fecal N to the suburban stream. Plants from a montane stream in a largely undeveloped recreational canyon that permitted off-leash dogs (δ15N = 1.8 ‰) were enriched relative to an adjacent canyon with similar land use that strictly prohibited dogs but had comparable vehicle traffic (δ15N = -0.7 ‰), suggesting the contribution of dog waste to stream N. Plants from urban stream reaches were enriched by 1.3 - 2.8 ‰ relative to upstream wildland reaches, and δ15N increased by 0.2 ‰ per km in the urban streams. Mechanisms leading to this urban enrichment could include leaky municipal sewers, atmospheric N deposition, and/or increased rates of N cycling and gaseous losses. Overall, our results demonstrate the potential utility of riparian plant N isotopes as a simple diagnostic of N source inputs to inform watershed management.

Identifying fecal matter contamination in produce fields using multispectral reflectance imaging under ambient solar illumination

NASA Astrophysics Data System (ADS)

Everard, Colm D.; Kim, Moon S.; Lee, Hoonsoo; O'Donnell, Colm P.

2016-05-01

An imaging device to detect fecal contamination in fresh produce fields could allow the producer avoid harvesting fecal contaminated produce. E.coli O157:H7 outbreaks have been associated with fecal contaminated leafy greens. In this study, in-field spectral profiles of bovine fecal matter, soil, and spinach leaves are compared. A common aperture imager designed with two identical monochromatic cameras, a beam splitter, and optical filters was used to simultaneously capture two-spectral images of leaves contaminated with both fecal matter and soil. The optical filters where 10 nm full width half maximum bandpass filters, one at 690 nm and the second at 710 nm. These were mounted in front of the object lenses. New images were created using the ratio of these two spectral images on a pixel by pixel basis. Image analysis results showed that the fecal matter contamination could be distinguished from soil and leaf on the ratio images. The use of this technology has potential to allow detection of fecal contamination in produce fields which can be a source of foodbourne illnesses. It has the added benefit of mitigating cross-contamination during harvesting and processing.

Survival of Salmonella adelaide and fecal coliforms in coarse sands of the swan costal plain, Western Australia.

PubMed Central

Parker, W F; Mee, B J

1982-01-01

The survival of Salmonella adelaide and fecal coliforms in two coarse sands influenced by two sources of septic tank effluent was studied. The experiments were conducted in conditions that reflected the soil environment beneath functioning septic tank systems. Significant differences in survival were found with different effluent sources. In one experiment the survival of S. adelaide was similar to that of fecal coliforms; in the other it was not. The nonuniform, multiphasic nature of survival curves was variability observed in these experiments suggests that the application of such survival data for establishing management criteria for septic tank systems--by, for example, the use of soil moisture characteristic curves to give estimates of movement in the soil--is inappropriate. PMID:7103482

Examination of the Source and Extended Virulence Genotypes of Escherichia coli Contaminating Retail Poultry Meat

PubMed Central

Johnson, Timothy J.; Logue, Catherine M.; Wannemuehler, Yvonne; Kariyawasam, Subhashinie; Doetkott, Curt; DebRoy, Chitrita; White, David G.

2009-01-01

Abstract Extraintestinal pathogenic Escherichia coli (ExPEC) are major players in human urinary tract infections, neonatal bacterial meningitis, and sepsis. Recently, it has been suggested that there might be a zoonotic component to these infections. To determine whether the E. coli contaminating retail poultry are possible extraintestinal pathogens, and to ascertain the source of these contaminants, they were assessed for their genetic similarities to E. coli incriminated in colibacillosis (avian pathogenic E. coli [APEC]), E. coli isolated from multiple locations of apparently healthy birds at slaughter, and human ExPEC. It was anticipated that the retail poultry isolates would most closely resemble avian fecal E. coli since only apparently healthy birds are slaughtered, and fecal contamination of carcasses is the presumed source of meat contamination. Surprisingly, this supposition proved incorrect, as the retail poultry isolates exhibited gene profiles more similar to APEC than to fecal isolates. These isolates contained a number of ExPEC-associated genes, including those associated with ColV virulence plasmids, and many belonged to the B2 phylogenetic group, known to be virulent in human hosts. Additionally, E. coli isolated from the crops and gizzards of apparently healthy birds at slaughter also contained a higher proportion of ExPEC-associated genes than did the avian fecal isolates examined. Such similarities suggest that the widely held beliefs about the sources of poultry contamination may need to be reassessed. Also, the presence of ExPEC-like clones on retail poultry meat means that we cannot yet rule out poultry as a source of ExPEC human disease. PMID:19580453

Clostridium difficile Infection and Fecal Microbiota Transplant

PubMed Central

Liubakka, Alyssa; Vaughn, Byron P.

2017-01-01

Clostridium difficile infection (CDI) is a major source of morbidity and mortality for hospitalized patients. Although most patients have a clinical response to existing antimicrobial therapies, recurrent infection develops in up to 30% of patients. Fecal microbiota transplant is a novel approach to this complex problem, with an efficacy rate of nearly 90% in the setting of multiple recurrent CDI. This review covers the current epidemiology of CDI (including toxigenic and nontoxigenic strains, risk factors for infection, and recurrent infection), methods of diagnosis, existing first-line therapies in CDI, the role of fecal microbiota transplant for multiple recurrent CDIs, and the potential use of fecal microbial transplant for patients with severe or refractory infection. PMID:27959316

Fecal Contamination on Produce from Wholesale and Retail Food Markets in Dhaka, Bangladesh.

PubMed

Harris, Angela R; Islam, Mohammad Aminul; Unicomb, Leanne; Boehm, Alexandria B; Luby, Stephen; Davis, Jennifer; Pickering, Amy J

2018-01-01

Fresh produce items can become contaminated with enteric pathogens along the supply chain at the preharvest (e.g., irrigation water, soil, fertilizer) or postharvest (e.g., vendor handling or consumer handling) stages. This study assesses the concentrations of fecal indicator bacteria Escherichia coli , enterococci (ENT), and Bacteriodales on surfaces of carrots, eggplants, red amaranth leaves, and tomatoes obtained from both a wholesale market (recently harvested) and neighborhood retail markets in Dhaka, Bangladesh. We detected E. coli in 100% of carrot and red amaranth rinses, 92% of eggplant rinses, and 46% of tomato rinses. Using a molecular microbial source tracking assay, we found that 32% of produce samples were positive for ruminant fecal contamination. Fecal indicator bacteria were more likely to be detected on produce collected in retail markets compared with that in the wholesale market; retail market produce were 1.25 times more likely to have E. coli detected ( P = 0.03) and 1.24 times more likely to have ENT detected ( P = 0.03) as compared with wholesale market produce. Bacteriodales was detected in higher concentrations in retail market produce samples compared with wholesale market produce samples (0.40 log 10 gene copies per 100 cm 2 higher, P = 0.03). Our results suggest that ruminant and general fecal contamination of produce in markets in Dhaka is common, and suggest that unsanitary conditions in markets are an important source of produce fecal contamination postharvest.

Arcobacter in Lake Erie beach waters: an emerging gastrointestinal pathogen linked with human-associated fecal contamination.

PubMed

Lee, Cheonghoon; Agidi, Senyo; Marion, Jason W; Lee, Jiyoung

2012-08-01

The genus Arcobacter has been associated with human illness and fecal contamination by humans and animals. To better characterize the health risk posed by this emerging waterborne pathogen, we investigated the occurrence of Arcobacter spp. in Lake Erie beach waters. During the summer of 2010, water samples were collected 35 times from the Euclid, Villa Angela, and Headlands (East and West) beaches, located along Ohio's Lake Erie coast. After sample concentration, Arcobacter was quantified by real-time PCR targeting the Arcobacter 23S rRNA gene. Other fecal genetic markers (Bacteroides 16S rRNA gene [HuBac], Escherichia coli uidA gene, Enterococcus 23S rRNA gene, and tetracycline resistance genes) were also assessed. Arcobacter was detected frequently at all beaches, and both the occurrence and densities of Arcobacter spp. were higher at the Euclid and Villa Angela beaches (with higher levels of fecal contamination) than at the East and West Headlands beaches. The Arcobacter density in Lake Erie beach water was significantly correlated with the human-specific fecal marker HuBac according to Spearman's correlation analysis (r = 0.592; P < 0.001). Phylogenetic analysis demonstrated that most of the identified Arcobacter sequences were closely related to Arcobacter cryaerophilus, which is known to cause gastrointestinal diseases in humans. Since human-pathogenic Arcobacter spp. are linked to human-associated fecal sources, it is important to identify and manage the human-associated contamination sources for the prevention of Arcobacter-associated public health risks at Lake Erie beaches.

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.

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

Occurrence of bacteria in Blue Marsh Lake and selected tributaries, Berks County, Pennsylvania; September-October 2001

USGS Publications Warehouse

Zimmerman, Michele L.

2002-01-01

The Commonwealth of Pennsylvania has water-quality standards that limit the number of specific bacteria in water that is considered safe for recreational use. Bacteria such as fecal streptococci, fecal coliforms, and Escherichia coli (E. coli) are used to assess recreational water quality because they usually live in the intestines of warm-blooded animals. Fecal indicator bacteria commonly are associated with waterborne disease-causing organisms (pathogens). These indicator bacteria are used routinely as a measure of the quality of water for recreational activities such as swimming, boating, and water skiing. If the indicator bacteria are present, effective measures could be taken to prevent the transmission or epidemic outbreak of waterborne diseases as a result of contamination of these waters from human or animal waste.Blue Marsh Lake is on Tulpehocken Creek in Berks County, Pa., and drains a largely agricultural basin. Land use in the basin is approximately 60 percent cropland, and 85 percent of the farms are livestock and poultry farms.The potential sources of fecal bacteria are:geese that inhabit the recreational areas of the lake,humans that visit the Dry Brooks Day Use Area (swimming area), andfarm animals, wastewater facilities, and household septic systems in the basin (bacteria from these sources could enter the lake through tributaries).To meet the recreational water-quality standard, lake water may not have more than 200 colony-forming units (CFU) of fecal coliforms per 100 milliliters (mL). During the week of July 23, 2001, data collected by the U.S. Army Corps of Engineers (USACE) at the swimming area at Blue Marsh Lake showed concentrations of fecal coliforms in the water exceeding the standard. To determine the extent of elevated concentrations of fecal indicator bacteria, further study of the lake and selected tributaries was needed.

[Nuclear magnetic resonance of anorectal malformations and persistent postoperative fecal incontinence].

PubMed

de Agustín, J C; Alami, H; Lassaletta, L; Gámez, M; Fernández, A; Fraile, E; Alenda, J G; Rollán, V; Utrilla, J G

1992-07-01

We review our experience with Magnetic Resonance Imaging (MRI) in the evaluation of 6 patients showing anorectal malformation, and 4 more with persistent postoperative fecal incontinence. Preoperative sagittal, axial and coronal planes were studied with special consideration to the pelvic and vertebral structures. The excellent resolution of MRI allowed accurate identification of the pelvic musculature in all patients, including those with bizarre sacral abnormalities. MRI revealed structural anomalies not detected previously, such as teathering cord, intraspinal lipoma, presacral mass and renal malformation. In our institution, MRI has replaced the CT scan in the study of patients suffering of persistent fecal incontinence. In non operated on cases of anorectal malformations, MRI determines with extraordinary accuracy the location of the rectal atretic pouch, the actual pelvic muscular quality, and the detection of previously unsuspected associated anomalies.

Sources and Pathways of Bacterial Contamination in Urban Streams and Ocean Beaches, Santa Barbara, California

NASA Astrophysics Data System (ADS)

Johnson, R. D.; Mendez, G. O.; La, J. X.; Izbicki, J. A.

2005-12-01

Streams and ocean beaches in Santa Barbara, California, occasionally have concentrations of fecal indicator bacteria that exceed public health standards for recreational water, forcing temporary beach closures. Possible sources of fecal bacteria contamination include transient human populations, animal populations, and leaking sewer lines. The purpose of this three-year study is to identify important sources of fecal bacteria affecting the urban streams and beaches and to identify important pathways of transport. Contamination may enter streams and beaches directly by surface runoff, but also may be transmitted short distances through shallow ground water. Our analysis of existing historical data shows that fecal indicator bacteria concentrations are higher in near-shore ocean water following extreme high tides. The possible role of near shore ground water in supplying contaminants to the sea will be investigated by sampling water from an array of shallow wells installed for this study between an older city sewer line and the ocean. The ground water flux to the ocean will be inferred from water levels in these wells, and further tested by radium isotope values in near shore ocean samples. Two additional well arrays will be installed to test for leakage from residential sewage hookups and measure associated exchanges between ground water, streams, and ocean. Preliminary data collected by this study show fecal indicator bacteria concentrations in urban reaches of Mission Creek and its tributaries, the principle drainage through the city, are higher during low flow periods than during periods of higher flow. Analysis of preliminary data also shows short-term temporal variations in bacterial concentrations during twenty-four hour periods. Human enterovirus has been detected in our sample from one urban-drain tributary to Mission Creek. In order to identify the origins of fecal indicator bacteria water samples from Mission Creek, its tributaries, urban drains, and associated shallow ground water will be analyzed for nutrients, dissolved organic carbon (including optical properties to characterize the composition of the organic carbon), fecal sterols, DNA (using Terminal-Restriction Fragment Length Poylmorphism), and phospholipid fatty acids.

Fecal parasite identification by microscopy and PCR in scimitar-horned oryx, Oryx dammah, managed at two sites.

PubMed

Pauling, Cassandra Dawn; Oller, Anna R; Jackson, Victoria

2016-12-01

The scimitar-horned oryx, Oryx dammah , an endangered species extinct in the wild, is managed in various captive management programs and is the focus of reintroduction efforts. Management variability can contribute to substantial parasite load differences, which can affect deworming programs and potentially transfer parasites to different regions with translocations. Parasite studies in O. dammah are lacking. In this study, we determined fecal egg/oocyst counts of O. dammah in two captive herds, Fossil Rim Wildlife Center (FRWC) and Kansas City Zoo (KCZ). Fecal egg counts (FEC) were performed on O. dammah feces collected seasonally using the modified McMaster method, and microscopy provided additional identification of parasite genera ova and oocysts. To identify parasites to species level, homogenized fecals provided DNA subjected to the polymerase chain reaction (PCR) using genus specific primers. Microscopy and sequencing results indicated the presence of Strongylus (Strongylus vulgaris, Angiostrongylus cantonensis) , Trichostrongylus (Haemonchus contortus, Camelostrongylus mentulatus) , Trichuris (T. leporis, T. ovis, and T. discolor) , Isospora (Isospora gryphoni) and Eimeria (E. zuernii and E. bovis) , with Strongylus being the most common. Nematodirus was identified through microscopy at FRWC. Fecal egg counts were significantly higher in (FRWC) than in (KCZ) in all samplings (P = <0.001). No significant difference was seen between parasite load and seasons (P = 0.103), nor site and season (P = 0.51). Both study sites maintained most animals within commonly accepted FEC levels found in domestic livestock. Individuals with high numbers of EPG or OPG were subordinate males, pregnant females, or neonates. Several significant interactions were found between genera of parasites, age, sex, season, and pregnancy status in the FRWC herd. Sampling limitations prevented further analysis of the KCZ herd. Understanding interactions between parasite loads and physiological, environmental, and regional differences can help determine inter-specific transfer of parasites, and establish appropriate anthelmintic programs for O. dammah herds.

TRANSPORT OF CHEMICAL AND MICROBIAL COMPOUNDS FROM KNOWN WASTEWATER DISCHARGES: POTENTIAL FOR USE AS INDICATORS OF HUMAN FECAL CONTAMINATION

EPA Science Inventory

The quality of drinking and recreational water currently (2005) is determined using indicator bacteria. However, the culture tests used to analyze for these bacteria require a long time to complete, and do not discriminate between human and animal fecal material sources. One co...

ASSESSMENT OF FECAL POLLUTION SOURCES IN PLUM CREEK WATERSHED USING BACTEROIDETES 16S RDNA-BASED ASSAYS

EPA Science Inventory

Recently, 16S rDNA Bacteroidetes-targeted PCR assays were developed to discriminate between ruminant and human fecal pollution. These assays are rapid and relatively inexpensive but have been used in a limited number of studies. In this study, we evaluated the efficacy o...

Detection limits and cost comparisons of human- and gull-associated conventional and quantitative PCR assays in artificial and environmental waters

EPA Science Inventory

Modern techniques for tracking fecal pollution in environmental waters require investing in DNA-based methods to determine the presence of specific fecal sources. To help water quality managers decide whether to employ routine polymerase chain reaction (PCR) or quantitative PC...

Multi-laboratory evaluations of the performance of Catellicoccus marimammalium PCR assays developed to target gull fecal sources

EPA Science Inventory

Here we report results from a multi-laboratory (n=11) evaluation of four different PCR methods targeting the 16S rRNA gene of Catellicoccus marimammalium used to detect fecal contamination from birds in coastal environments. The methods included conventional end-point PCR, a SYBR...

Comparison of two poultry litter qPCR assays targeting the 16S rRNA gene of Brevibacterium sp

EPA Science Inventory

Chicken feces are vectors of human pathogens and are also important sources of fecal pollution in environmental waters. Consequently, methods that can detect chicken fecal pollution are needed in public health and environmental monitoring studies. In this study, we compared a pre...

TURKEY FECAL MICROBIAL COMMUNITY STRUCTURE AND ECOLOGICAL FUNCTIONS REVEALED BY 16S RDNA AND METAGENOME SEQUENCES

EPA Science Inventory

Turkey feces are an important source of fecal waste in the United States. With the exception of isolated studies on bacterial pathogens, little is known about the type of bacteria inhabiting the turkey gut. In order to understand the microbial diversity and functional genes assoc...

The potential of gut microbiota and fecal volatile organic compounds analysis as early diagnostic biomarker for necrotizing enterocolitis and sepsis in preterm infants.

PubMed

Berkhout, Daniel Johannes Cornelis; Niemarkt, Hendrik Johannes; de Boer, Nanne Klaas Hendrik; Benninga, Marc Alexander; de Meij, Timotheüs Gualtherus Jacob

2018-05-01

Although the exact pathophysiological mechanisms of both necrotizing enterocolitis (NEC) and late-onset sepsis (LOS) in preterm infants are yet to be elucidated, evidence is emerging that the gut microbiota plays a key role in their pathophysiology. Areas covered: In this review, initial microbial colonization and factors influencing microbiota composition are discussed. For both NEC and LOS, an overview of studies investigating preclinical alterations in gut microbiota composition and fecal volatile organic compounds (VOCs) is provided. Fecal VOCs are considered to reflect not only gut microbiota composition, but also their metabolic activity and concurrent interaction with the host. Expert review: Heterogeneity in study protocols and applied analytical techniques hampers reliable comparison between outcomes of different microbiota studies, limiting the ability to draw firm conclusions. This dilemma is illustrated by the finding that study results often cannot be reproduced, or even contradict each other. A NEC- and sepsis specific microbial or metabolic signature has not yet been discovered. Identification of 'disease-specific' VOCs and microbiota composition may increase understanding on pathophysiological mechanisms and may allow for development of an accurate screening tool, opening avenues towards timely identification and initiation of targeted treatment for preterm infants at increased risk for NEC and sepsis.

Evaluation of multiplex tandem real-time PCR for detection of Cryptosporidium spp., Dientamoeba fragilis, Entamoeba histolytica, and Giardia intestinalis in clinical stool samples.

PubMed

Stark, D; Al-Qassab, S E; Barratt, J L N; Stanley, K; Roberts, T; Marriott, D; Harkness, J; Ellis, J T

2011-01-01

The aim of this study was to describe the first development and evaluation of a multiplex tandem PCR (MT-PCR) assay for the detection and identification of 4 common pathogenic protozoan parasites, Cryptosporidium spp., Dientamoeba fragilis, Entamoeba histolytica, and Giardia intestinalis, from human clinical samples. A total of 472 fecal samples submitted to the Department of Microbiology at St. Vincent's Hospital were included in the study. The MT-PCR assay was compared to four real-time PCR (RT-PCR) assays and microscopy by a traditional modified iron hematoxylin stain. The MT-PCR detected 28 G. intestinalis, 26 D. fragilis, 11 E. histolytica, and 9 Cryptosporidium sp. isolates. Detection and identification of the fecal protozoa by MT-PCR demonstrated 100% correlation with the RT-PCR results, and compared to RT-PCR, MT-PCR exhibited 100% sensitivity and specificity, while traditional microscopy of stained fixed fecal smears exhibited sensitivities and specificities of 56% and 100% for Cryptosporidium spp., 38% and 99% for D. fragilis, 47% and 97% for E. histolytica, and 50% and 100% for G. intestinalis. No cross-reactivity was detected in 100 stool samples containing various other bacterial, viral, and protozoan species. The MT-PCR assay was able to provide rapid, sensitive, and specific simultaneous detection and identification of the four most important diarrhea-causing protozoan parasites that infect humans. This study also highlights the lack of sensitivity demonstrated by microscopy, and thus, molecular methods such as MT-PCR must be considered the diagnostic methods of choice for enteric protozoan parasites.

Elevated fecal peptidase D at onset of colitis in Galphai2-/- mice, a mouse model of IBD.

PubMed

Bergemalm, Daniel; Kruse, Robert; Sapnara, Maria; Halfvarson, Jonas; Hörnquist, Elisabeth Hultgren

2017-01-01

The identification of novel fecal biomarkers in inflammatory bowel disease (IBD) is hampered by the complexity of the human fecal proteome. On the other hand, in experimental mouse models there is probably less variation. We investigated the fecal protein content in mice to identify possible biomarkers and pathogenic mechanisms. Fecal samples were collected at onset of inflammation in Galphai2-/- mice, a well-described spontaneous model of chronic colitis, and from healthy littermates. The fecal proteome was analyzed by two-dimensional electrophoresis and quantitative mass spectrometry and results were then validated in a new cohort of mice. As a potential top marker of disease, peptidase D was found at a higher ratio in Galphai2-/- mouse feces relative to controls (fold change 27; p = 0.019). Other proteins found to be enriched in Gαi2-/- mice were mainly pancreatic proteases, and proteins from plasma and blood cells. A tendency of increased calprotectin, subunit S100-A8, was also observed (fold change 21; p = 0.058). Proteases are potential activators of inflammation in the gastrointestinal tract through their interaction with the proteinase-activated receptor 2 (PAR2). Accordingly, the level of PAR2 was found to be elevated in both the colon and the pancreas of Galphai2-/- mice at different stages of disease. These findings identify peptidase D, an ubiquitously expressed intracellular peptidase, as a potential novel marker of colitis. The elevated levels of fecal proteases may be involved in the pathogenesis of colitis and contribute to the clinical phenotype, possibly by activation of intestinal PAR2.

Fecal contamination of drinking-water in low- and middle-income countries: a systematic review and meta-analysis.

PubMed

Bain, Robert; Cronk, Ryan; Wright, Jim; Yang, Hong; Slaymaker, Tom; Bartram, Jamie

2014-05-01

Access to safe drinking-water is a fundamental requirement for good health and is also a human right. Global access to safe drinking-water is monitored by WHO and UNICEF using as an indicator "use of an improved source," which does not account for water quality measurements. Our objectives were to determine whether water from "improved" sources is less likely to contain fecal contamination than "unimproved" sources and to assess the extent to which contamination varies by source type and setting. Studies in Chinese, English, French, Portuguese, and Spanish were identified from online databases, including PubMed and Web of Science, and grey literature. Studies in low- and middle-income countries published between 1990 and August 2013 that assessed drinking-water for the presence of Escherichia coli or thermotolerant coliforms (TTC) were included provided they associated results with a particular source type. In total 319 studies were included, reporting on 96,737 water samples. The odds of contamination within a given study were considerably lower for "improved" sources than "unimproved" sources (odds ratio [OR] = 0.15 [0.10-0.21], I2 = 80.3% [72.9-85.6]). However over a quarter of samples from improved sources contained fecal contamination in 38% of 191 studies. Water sources in low-income countries (OR = 2.37 [1.52-3.71]; p<0.001) and rural areas (OR = 2.37 [1.47-3.81] p<0.001) were more likely to be contaminated. Studies rarely reported stored water quality or sanitary risks and few achieved robust random selection. Safety may be overestimated due to infrequent water sampling and deterioration in quality prior to consumption. Access to an "improved source" provides a measure of sanitary protection but does not ensure water is free of fecal contamination nor is it consistent between source types or settings. International estimates therefore greatly overstate use of safe drinking-water and do not fully reflect disparities in access. An enhanced monitoring strategy would combine indicators of sanitary protection with measures of water quality.

IDENTIFICATION OF AVIAN-SPECIFIC FECAL METAGENOMIC SEQUENCES USING GENOME FRAGMENT ENRICHMENTS

EPA Science Inventory

Sequence analysis of microbial genomes has provided biologists the opportunity to compare genetic differences between closely related microorganisms. While random sequencing has also been used to study natural microbial communities, metagenomic comparisons via sequencing analysis...

Recurrence of fecal coliforms and Salmonella species in biosolids following thermophilic anaerobic digestion.

PubMed

Iranpour, Reza; Cox, Huub H J

2006-09-01

The U.S. Environmental Protection Agency (U.S. EPA) Part 503 Biosolids Rule requires the fecal coliform (indicator) or Salmonella species (pathogen) density requirements for Class A biosolids to be met at the last point of plant control (truck-loading facility and/or farm for land application). The three Southern Californian wastewater treatment plants in this study produced biosolids by thermophilic anaerobic digestion and all met the Class A limits for both fecal coliforms and Salmonella sp. in the digester outflow biosolids. At two plants, however, a recurrence of fecal coliforms was observed in postdigestion biosolids, which caused exceedance of the Class A limit for fecal coliforms at the truck-loading facility and farm for land application. Comparison of observations at the three plants and further laboratory tests indicated that the recurrence of fecal coliforms can possibly be related to the following combination of factors: (1) incomplete destruction of fecal coliforms during thermophilic anaerobic digestion, (2) contamination of Class A biosolids with fecal coliforms from external sources during postdigestion, (3) a large drop of the postdigestion biosolids temperature to below the maximum for fecal coliform growth, (4) an unknown effect of biosolids dewatering in centrifuges. At Hyperion Treatment Plant (City of Los Angeles, California), fecal coliform recurrence could be prevented by the following: (1) complete conversion to thermophilic operation to exclude contamination by mesophilically digested biosolids and (2) insulation and electrical heat-tracing of postdigestion train for maintaining a high biosolids temperature in postdigestion.

Storm loads of culturable and molecular fecal indicators in an inland urban stream.

PubMed

Liao, Hehuan; Krometis, Leigh-Anne H; Cully Hession, W; Benitez, Romina; Sawyer, Richard; Schaberg, Erin; von Wagoner, Emily; Badgley, Brian D

2015-10-15

Elevated concentrations of fecal indicator bacteria in receiving waters during wet-weather flows are a considerable public health concern that is likely to be exacerbated by future climate change and urbanization. Knowledge of factors driving the fate and transport of fecal indicator bacteria in stormwater is limited, and even less is known about molecular fecal indicators, which may eventually supplant traditional culturable indicators. In this study, concentrations and loading rates of both culturable and molecular fecal indicators were quantified throughout six storm events in an instrumented inland urban stream. While both concentrations and loading rates of each fecal indicator increased rapidly during the rising limb of the storm hydrographs, it is the loading rates rather than instantaneous concentrations that provide a better estimate of transport through the stream during the entire storm. Concentrations of general fecal indicators (both culturable and molecular) correlated most highly with each other during storm events but not with the human-associated HF183 Bacteroides marker. Event loads of general fecal indicators most strongly correlated with total runoff volume, maximum discharge, and maximum turbidity, while event loads of HF183 most strongly correlated with the time to peak flow in a hydrograph. These observations suggest that collection of multiple samples during a storm event is critical for accurate predictions of fecal indicator loading rates and total loads during wet-weather flows, which are required for effective watershed management. In addition, existing predictive models based on general fecal indicators may not be sufficient to predict source-specific genetic markers of fecal contamination. Copyright © 2015 Elsevier B.V. All rights reserved.

Rapid QPCR-based assay for fecal Bacteroides spp. as a tool for assessing fecal contamination in recreational waters.

PubMed

Converse, Reagan R; Blackwood, A Denene; Kirs, Marek; Griffith, John F; Noble, Rachel T

2009-11-01

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

Using fecal sterols to assess dynamics of sewage input in sediments along a human-impacted river-estuary system in eastern China.

PubMed

He, Ding; Zhang, Kai; Tang, Jianhui; Cui, Xingqian; Sun, Yongge

2018-05-01

Sedimentary fecal sterols and other sterol biomarkers, combined with bulk total organic carbon (TOC) and its stable carbon isotope were applied to characterize the sewage contamination across a ca. 280 km transect from the Xiaoqing River to the Laizhou Bay, a typical river-estuary system subjected to extensive anthropogenic stress due to rapid regional urbanization and industrialization in eastern China. Two sampling events were performed in both spring and summer seasons in the Laizhou Bay adjacent to the Xiaoqing River in order to assess the potential seasonal variation. Fecal sterols such as coprostanol and epicoprostanol, which are typical indicators of anthropogenic sewage input, displayed high concentrations of up to 63.2 μg g -1 dry weight (dw) and 13.1 μg g -1 dw, respectively. Results suggested that most of the stations along the Xiaoqing River were severely contaminated by fecal inputs with a decreasing trend from the river to the estuary that was mainly explained by the increasing distance from the diffuse sewage sources and the gradual dilution by sea water. Although there was no significant difference in fecal sterol concentrations between spring and summer in the Laizhou Bay, suggestive of no significant difference in sewage abundance, significantly higher average epicoprostanol/coprostanol and lower coprostanol/epicoprostanol ratios were observed in spring than summer, indicative of different sewage sources (e.g., human vs. non-human). Seasonal discharge and land-runoff, air temperature related to microbial activity differences and different extend of animal manure irrigation during agricultural planting could be additional reasons and need further investigation. Nevertheless, fecal sterol concentrations, distributions and diagnostic ratios should all be taken into consideration to better understand sewage inputs and source dynamics in river-estuary ecosystems. Copyright © 2018 Elsevier B.V. All rights reserved.

Fiber supplementation influences phylogenetic structure and functional capacity of the human intestinal microbiome: follow-up of a randomized controlled trial.

PubMed

Holscher, Hannah D; Caporaso, J Gregory; Hooda, Seema; Brulc, Jennifer M; Fahey, George C; Swanson, Kelly S

2015-01-01

In our published randomized, double-blind, placebo-controlled, 3-period crossover trial, healthy adult men (n = 21) consumed bars containing no supplemental fiber (placebo; NFC), polydextrose (21 g/d), and soluble corn fiber (SCF; 21 g/d) for 21 d each. Fecal specimens were collected between days 16 and 21 for fermentative end-product analysis and 16S ribosomal RNA bacterial gene amplification for bacterial taxa identification. Fiber supplementation decreased fecal putrefaction compounds and shifted abundances of several bacterial taxa. The objective was to perform whole-genome shotgun 454 pyrosequencing on the same fecal specimens collected in that clinical trial to obtain comprehensive fecal bacterial genome sequencing coverage and explore the full range of bacterial genetic information in the fecal microbiome, thereby using a systematic approach to study the impact of dietary fiber supplementation on fecal metabolites, bacterial taxa, and bacterial metagenomes. Fecal samples were subjected to whole-genome shotgun 454 pyrosequencing to identify both fecal bacterial populations present and their functional genetic capacity. Whole-genome shotgun sequencing results revealed that fiber consumption shifted the Bacteroidetes:Firmicutes ratio, increasing the relative abundance of Bacteroidetes 12 ± 2% and 13 ± 2% with polydextrose and SCF, respectively, compared with NFC. Bivariate correlations showed a positive correlation between the Bacteroidetes:Firmicutes ratio and total dietary fiber intake but not body mass index. Principal coordinates analysis of Bray-Curtis distances indicated that bacterial gene composition was more similar in participants consuming fibers (polydextrose and SCF combined) in comparison with NFC. Shifts in bacterial gene abundances after polydextrose and SCF supplementation included genes associated with carbohydrate, amino acid, and lipid metabolism, as well as metabolism of cofactors and vitamins. This study conveys novel information about the impact of dietary fiber supplementation on the phylogenetic structure and functional capacity of the fecal microbiome of healthy adults. © 2015 American Society for Nutrition.

Integral strategy for evaluation of fecal indicator performance in bird-influenced saline inland waters.

PubMed

Kirschner, Alexander K T; Zechmeister, Thomas C; Kavka, Gerhard G; Beiwl, Christian; Herzig, Alois; Mach, Robert L; Farnleitner, Andreas H

2004-12-01

Wild birds are an important nonpoint source of fecal contamination of surface waters, but their contribution to fecal pollution is mostly difficult to estimate. Thus, to evaluate the relation between feces production and input of fecal indicator bacteria (FIB) into aquatic environments by wild waterfowl, we introduced a new holistic approach for evaluating the performance of FIB in six shallow saline habitats. For this, we monitored bird abundance, fecal pellet production, and the abundance of FIB concomitantly with a set of environmental variables over a 9-month period. For estimating fecal pellet production, a new protocol of fecal pellet counting was introduced, which was called fecal taxation (FTX). We could show that, over the whole range of investigated habitats, bird abundance, FTX values, and FIB abundance were highly significantly correlated and could demonstrate the good applicability of the FTX as a meaningful surrogate parameter for recent bird abundances and fecal contamination by birds in shallow aquatic ecosystems. Presumptive enterococci (ENT) were an excellent surrogate parameter of recent fecal contamination in these saline environments for samples collected at biweekly to monthly sampling intervals while presumptive Escherichia coli and fecal coliforms (FC) were often undetectable. Significant negative correlations with salinity indicated that E. coli and FC survival was hampered by osmotic stress. Statistical analyses further revealed that fecal pollution-associated parameters represented one system component independent from other environmental variables and that, besides feces production, rainfall, total suspended solids (direct), and trophy (indirect) had significant positive effects on ENT concentrations. Our holistic approach of linking bird abundance, feces production, and FIB detection with environmental variables may serve as a powerful model for application to other aquatic ecosystems.

Integral Strategy for Evaluation of Fecal Indicator Performance in Bird-Influenced Saline Inland Waters

PubMed Central

Kirschner, Alexander K. T.; Zechmeister, Thomas C.; Kavka, Gerhard G.; Beiwl, Christian; Herzig, Alois; Mach, Robert L.; Farnleitner, Andreas H.

2004-01-01

Wild birds are an important nonpoint source of fecal contamination of surface waters, but their contribution to fecal pollution is mostly difficult to estimate. Thus, to evaluate the relation between feces production and input of fecal indicator bacteria (FIB) into aquatic environments by wild waterfowl, we introduced a new holistic approach for evaluating the performance of FIB in six shallow saline habitats. For this, we monitored bird abundance, fecal pellet production, and the abundance of FIB concomitantly with a set of environmental variables over a 9-month period. For estimating fecal pellet production, a new protocol of fecal pellet counting was introduced, which was called fecal taxation (FTX). We could show that, over the whole range of investigated habitats, bird abundance, FTX values, and FIB abundance were highly significantly correlated and could demonstrate the good applicability of the FTX as a meaningful surrogate parameter for recent bird abundances and fecal contamination by birds in shallow aquatic ecosystems. Presumptive enterococci (ENT) were an excellent surrogate parameter of recent fecal contamination in these saline environments for samples collected at biweekly to monthly sampling intervals while presumptive Escherichia coli and fecal coliforms (FC) were often undetectable. Significant negative correlations with salinity indicated that E. coli and FC survival was hampered by osmotic stress. Statistical analyses further revealed that fecal pollution-associated parameters represented one system component independent from other environmental variables and that, besides feces production, rainfall, total suspended solids (direct), and trophy (indirect) had significant positive effects on ENT concentrations. Our holistic approach of linking bird abundance, feces production, and FIB detection with environmental variables may serve as a powerful model for application to other aquatic ecosystems. PMID:15574941

Portable multispectral fluorescence imaging system for food safety applications

NASA Astrophysics Data System (ADS)

Lefcourt, Alan M.; Kim, Moon S.; Chen, Yud-Ren

2004-03-01

Fluorescence can be a sensitive method for detecting food contaminants. Of particular interest is detection of fecal contamination as feces is the source of many pathogenic organisms. Feces generally contain chlorophyll a and related compounds due to ingestion of plant materials, and these compounds can readily be detected using fluorescence techniques. Described is a fluorescence-imaging system consisting primarily of a UV light source, an intensified camera with a six-position filter wheel, and software for controlling the system and automatically analyzing the resulting images. To validate the system, orchard apples artificially contaminated with dairy feces were used in a "hands-on" public demonstration. The contamination sites were easily identified using automated edge detection and threshold detection algorithms. In addition, by applying feces to apples and then washing sets of apples at hourly intervals, it was determined that five h was the minimum contact time that allowed identification of the contamination site after the apples were washed. There are many potential uses for this system, including studying the efficacy of apple washing systems.

The effect of age and carbohydrate and protein sources on digestibility, fecal microbiota, fermentation products, fecal IgA, and immunological blood parameters in dogs.

PubMed

Maria, A P J; Ayane, L; Putarov, T C; Loureiro, B A; Neto, B P; Casagrande, M F; Gomes, M O S; Glória, M B A; Carciofi, A C

2017-06-01

The present study compared the effects of diets formulated with fibers of different fermentability and protein sources of animal or vegetable origins on old and adult dogs. The experiment was organized in a 3 (diets) × 2 (ages) factorial arrangement, totaling 6 treatments. Thirty-six Beagle dogs were used (18 old dogs [10.2 ± 1.0 yr] and 18 young adult dogs [2.6 ± 0.9 yr]), with 6 dogs per treatment. Three diets with similar compositions were used: a nonfermentable insoluble fiber source (sugarcane fiber) and chicken byproduct meal (nonfermentable fiber [NFF] diet), a fermentable fiber source (beet pulp) and chicken byproduct meal (fermentable fiber [FF] diet), and soybean meal as a protein and fiber source (soybean meal [SM] diet). Data were evaluated using the MIXED procedure and considering the effects and interactions of block, animal, diets, and age. Means were compared using Tukey's test ( < 0.05). Age × diet interactions were evaluated when < 0.1. Old dogs had a reduced coefficient of total tract apparent digestibility of DM, which was explained by the age and diet interaction of CP and fat digestibility that was lower for old than for adult dogs fed the FF diet ( < 0.05). The SM diet obtained higher DM, OM, CP, and fiber digestibility compared with the NFF diet ( < 0.05). The feces of dogs fed the NFF diet had increased DM content ( < 0.05). The short-chain fatty acids (SCFA) did not change by age group and were higher for dogs fed the FF and SM diets compared with dogs fed the NFF diet ( < 0.05). An age and diet interaction was observed for lactate and was increased in the feces of old dogs compared with adult dogs fed the FF diet ( < 0.05). Fecal putrescine, cadaverine, and spermine were increased for old dogs compared with adult dogs ( < 0.05), and the spermidine fecal concentration was increased for dogs fed the SM diet regardless of age ( < 0.05). Old dogs had reduced peripheral T and B lymphocytes ( < 0.05). An age and diet interaction was observed for fecal IgA ( < 0.001). Adult dogs fed the SM diet had increased IgA in feces compared with animals fed the NFF and FF diets ( < 0.05). However, for old dogs, both the FF and SM diets induced increased IgA compared with the NFF diet ( < 0.05). In conclusion, beet pulp may reduce digestibility and induce increased lactate in the feces of old dogs. The protein and oligosaccharides of soybean meal are digestible by dogs, induce the production of SCFA and spermidine, and increase fecal IgA. Old dogs had increased putrecine, cadaverine, and spermine fecal concentrations.

Microbial source tracking using host specific FAME profiles of fecal coliforms.

PubMed

Duran, Metin; Haznedaroğlu, Berat Z; Zitomer, Daniel H

2006-01-01

The objective of this study was to investigate the host-specific differences in fatty acid methyl ester (FAME) profiles of fecal coliforms (FC). A known-source library was constructed with 314 FC isolates cultured from 6 possible sources of fecal pollution; 99 isolates from sewage; 29 from bovine; 29 from poultry; 50 from swine; 46 from waterfowl; and 61 from deer. It was found that the hydroxy FAMEs 12:0 2 OH, 12:03 OH, and 14:02 OH were exclusively associated with isolates of human origin. On the other hand, 3 saturated FAMEs, 10:0, 15:0, and 18:0 were found only in isolates from non-human sources, 15:0 being associated with livestock samples only. In addition to the presence of these signature FAMEs, the mean relative masses of 16:1 omega7c and 16:1 ISO/14:03 OH were significantly different between the isolates of human and non-human origins. A linear discriminant function differentiated FC isolates of human origin from those of livestock and wildlife origin at 99% accuracy. These results strongly suggest that the FAME profiles of FC show statistically significant host specificity and may have the potential to be used as a phenotypic microbial source tracking tool.

Fiber source and inclusion level affects characteristics of excreta from growing pigs

PubMed Central

Ndou, Saymore Petros; Bakare, Archibold Garikayi

2018-01-01

Objective The objective of the study was to determine the influence of varying fibrous diets on fecal characteristics of growing pigs. Methods A total of 104 pigs (initial weight 18±2.0 kg) were used in the study. They were housed in individual pens and fed on diets containing maize cob, grass hay, lucerne hay, maize stover, and sunflower husk. These fibers were included at 0, 80, 160, 240, 320 and 400 g/kg. Fecal and urine samples were collected. Results Fecal output was largest amongst pigs fed on diets containing grass hay and maize stover (p<0.05). Nitrogen content was highest in feces from pigs fed on sunflower husk (p< 0.05). Pigs fed on diets containing maize stover and maize cobs produced the largest concentrations of short chain fatty acids. Acetate concentration was high in feces of pigs fed maize stover than those fed grass hay and lucerne hay (p<0.05). As the level of fiber inclusion increased, fecal consistency and nitrogen content increased linearly (p<0.05). Urea nitrogen decreased as the inclusion level increased across all the fibers (p<0.05), with maize cobs containing the largest content of urea nitrogen. As dietary fiber content increased, fecal nitrogen content also increased (p<0.05). Conclusion It was concluded that different fiber sources influence fecal characteristics, thereby having different implications on pig waste management. It is vital to monitor fiber inclusion thresholds so as to easily manage environmental pollutants such as butyrate that contribute to odors. PMID:26954189

Influence of seasonal and inter-annual hydro-meteorological variability on surface water fecal coliform concentration under varying land-use composition.

PubMed

St Laurent, Jacques; Mazumder, Asit

2014-01-01

Quantifying the influence of hydro-meteorological variability on surface source water fecal contamination is critical to the maintenance of safe drinking water. Historically, this has not been possible due to the scarcity of data on fecal indicator bacteria (FIB). We examined the relationship between hydro-meteorological variability and the most commonly measured FIB, fecal coliform (FC), concentration for 43 surface water sites within the hydro-climatologically complex region of British Columbia. The strength of relationship was highly variable among sites, but tended to be stronger in catchments with nival (snowmelt-dominated) hydro-meteorological regimes and greater land-use impacts. We observed positive relationships between inter-annual FC concentration and hydro-meteorological variability for around 50% of the 19 sites examined. These sites are likely to experience increased fecal contamination due to the projected intensification of the hydrological cycle. Seasonal FC concentration variability appeared to be driven by snowmelt and rainfall-induced runoff for around 30% of the 43 sites examined. Earlier snowmelt in nival catchments may advance the timing of peak contamination, and the projected decrease in annual snow-to-precipitation ratio is likely to increase fecal contamination levels during summer, fall, and winter among these sites. Safeguarding drinking water quality in the face of such impacts will require increased monitoring of FIB and waterborne pathogens, especially during periods of high hydro-meteorological variability. This data can then be used to develop predictive models, inform source water protection measures, and improve drinking water treatment. Copyright © 2013 Elsevier Ltd. All rights reserved.

Water quality associated public health risk in Bo, Sierra Leone.

PubMed

Jimmy, David H; Sundufu, Abu J; Malanoski, Anthony P; Jacobsen, Kathryn H; Ansumana, Rashid; Leski, Tomasz A; Bangura, Umaru; Bockarie, Alfred S; Tejan, Edries; Lin, Baochuan; Stenger, David A

2013-01-01

Human health depends on reliable access to safe drinking water, but in many developing countries only a limited number of wells and boreholes are available. Many of these water resources are contaminated with biological or chemical pollutants. The goal of this study was to examine water access and quality in urban Bo, Sierra Leone. A health census and community mapping project in one neighborhood in Bo identified the 36 water sources used by the community. A water sample was taken from each water source and tested for a variety of microbiological and physicochemical substances. Only 38.9% of the water sources met World Health Organization (WHO) microbial safety requirements based on fecal coliform levels. Physiochemical analysis indicated that the majority (91.7%) of the water sources met the requirements set by the WHO. In combination, 25% of these water resources met safe drinking water criteria. No variables associated with wells were statistically significant predictors of contamination. This study indicated that fecal contamination is the greatest health risk associated with drinking water. There is a need to raise hygiene awareness and implement inexpensive methods to reduce fecal contamination and improve drinking water safety in Bo, Sierra Leone.

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 ...

40 CFR 432.125 - New source performance standards (NSPS).

Code of Federal Regulations, 2010 CFR

2010-07-01

... BOD5 26 16 Fecal Coliform (2) (3) O&G (as HEM) 14 8.0 TSS 30 20 1 mg/L (ppm). 2 Maximum of 400 MPN or... Ammonia (as N) 8.0 4.0 BOD5 26 16 Fecal Coliform (2) (3) O&G (as HEM) 14 8.0 TSS 30 20 Total Nitrogen 147...

40 CFR 432.125 - New source performance standards (NSPS).

Code of Federal Regulations, 2011 CFR

2011-07-01

... BOD5 26 16 Fecal Coliform (2) (3) O&G (as HEM) 14 8.0 TSS 30 20 1 mg/L (ppm). 2 Maximum of 400 MPN or... Ammonia (as N) 8.0 4.0 BOD5 26 16 Fecal Coliform (2) (3) O&G (as HEM) 14 8.0 TSS 30 20 Total Nitrogen 147...

Effects of DNA Extraction Procedures on Bacteroides Profiles in Fecal Samples From Various Animals Determined by Terminal Restriction Fragment Length Polymorphism Analysis

EPA Science Inventory

A major assumption in microbial source tracking is that some fecal bacteria are specific to a host animal, and thus provide unique microbial fingerprints that can be used to differentiate hosts. However, the DNA information obtained from a particular sample may be biased dependi...

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...

NMR-based fecal metabolomics fingerprinting as predictors of earlier diagnosis in patients with colorectal cancer.

PubMed

Lin, Yan; Ma, Changchun; Liu, Chengkang; Wang, Zhening; Yang, Jurong; Liu, Xinmu; Shen, Zhiwei; Wu, Renhua

2016-05-17

Colorectal cancer (CRC) is a growing cause of mortality in developing countries, warranting investigation into its earlier detection for optimal disease management. A metabolomics based approach provides potential for noninvasive identification of biomarkers of colorectal carcinogenesis, as well as dissection of molecular pathways of pathophysiological conditions. Here, proton nuclear magnetic resonance spectroscopy (1HNMR) -based metabolomic approach was used to profile fecal metabolites of 68 CRC patients (stage I/II=20; stage III=25 and stage IV=23) and 32 healthy controls (HC). Pattern recognition through principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) was applied on 1H-NMR processed data for dimension reduction. OPLS-DA revealed that each stage of CRC could be clearly distinguished from HC based on their metabolomic profiles. Successive analyses identified distinct disturbances to fecal metabolites of CRC patients at various stages, compared with those in cancer free controls, including reduced levels of acetate, butyrate, propionate, glucose, glutamine, and elevated quantities of succinate, proline, alanine, dimethylglycine, valine, glutamate, leucine, isoleucine and lactate. These altered fecal metabolites potentially involved in the disruption of normal bacterial ecology, malabsorption of nutrients, increased glycolysis and glutaminolysis. Our findings revealed that the fecal metabolic profiles of healthy controls can be distinguished from CRC patients, even in the early stage (stage I/II), highlighting the potential utility of NMR-based fecal metabolomics fingerprinting as predictors of earlier diagnosis in CRC patients.

NMR-based fecal metabolomics fingerprinting as predictors of earlier diagnosis in patients with colorectal cancer

PubMed Central

Lin, Yan; Ma, Changchun; Liu, Chengkang; Wang, Zhening; Yang, Jurong; Liu, Xinmu; Shen, Zhiwei; Wu, Renhua

2016-01-01

Colorectal cancer (CRC) is a growing cause of mortality in developing countries, warranting investigation into its earlier detection for optimal disease management. A metabolomics based approach provides potential for noninvasive identification of biomarkers of colorectal carcinogenesis, as well as dissection of molecular pathways of pathophysiological conditions. Here, proton nuclear magnetic resonance spectroscopy (1HNMR) -based metabolomic approach was used to profile fecal metabolites of 68 CRC patients (stage I/II=20; stage III=25 and stage IV=23) and 32 healthy controls (HC). Pattern recognition through principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) was applied on 1H-NMR processed data for dimension reduction. OPLS-DA revealed that each stage of CRC could be clearly distinguished from HC based on their metabolomic profiles. Successive analyses identified distinct disturbances to fecal metabolites of CRC patients at various stages, compared with those in cancer free controls, including reduced levels of acetate, butyrate, propionate, glucose, glutamine, and elevated quantities of succinate, proline, alanine, dimethylglycine, valine, glutamate, leucine, isoleucine and lactate. These altered fecal metabolites potentially involved in the disruption of normal bacterial ecology, malabsorption of nutrients, increased glycolysis and glutaminolysis. Our findings revealed that the fecal metabolic profiles of healthy controls can be distinguished from CRC patients, even in the early stage (stage I/II), highlighting the potential utility of NMR-based fecal metabolomics fingerprinting as predictors of earlier diagnosis in CRC patients. PMID:27107423

Water quality in the Withers Swash Basin, with emphasis on enteric bacteria, Myrtle Beach, South Carolina, 1991-93

USGS Publications Warehouse

Guimaraes, W.B.

1995-01-01

Water samples were collected in 1991-93 from Withers Swash and its two tributaries (the Mainstem and KOA Branches) in Myrtle Beach, S.C., and analyzed for physical properties, organic and inorganic constituents, and fecal coliform and streptococcus bacteria. Samples were collected during wet- and dry-weather conditions to assess the water quality of the streams before and after storm runoff. Water samples were analyzed for over 200 separate physical, chemical, and biological constituents. Concentrations of 11 constituents violated State criteria for shellfish harvesting waters, and State Human Health Criteria. The 11 constituents included concentrations of dissolved oxygen, arsenic, lead, cadmium, mercury, chlordane, dieldrin, 1,1,1-trichloroethane, 1,1-dichloroethylene, trichloroethylene, and fecal coliform bacteria. Water samples were examined for the presence of enteric bacteria (fecal coliform and fecal streptococcus) at 46 sites throughout the Withers Swash Basin and 5 sites on the beach and in the Atlantic Ocean. Water samples were collected just upstream from all confluences in order to determine sources of bacterial contamination. Temporally and spatially high concentrations of enteric bacteria were detected throughout the Withers Swash Basin; however, these sporadic bacteria concentrations made it difficult to determine a single source of the contamination. These enteric bacteria concentrations are probably derived from a number of sources in the basin including septic tanks, garbage containers, and the feces of waterfowl and domestic animals.

Association of Supply Type with Fecal Contamination of Source Water and Household Stored Drinking Water in Developing Countries: A Bivariate Meta-analysis.

PubMed

Shields, Katherine F; Bain, Robert E S; Cronk, Ryan; Wright, Jim A; Bartram, Jamie

2015-12-01

Access to safe drinking water is essential for health. Monitoring access to drinking water focuses on water supply type at the source, but there is limited evidence on whether quality differences at the source persist in water stored in the household. We assessed the extent of fecal contamination at the source and in household stored water (HSW) and explored the relationship between contamination at each sampling point and water supply type. We performed a bivariate random-effects meta-analysis of 45 studies, identified through a systematic review, that reported either the proportion of samples free of fecal indicator bacteria and/or individual sample bacteria counts for source and HSW, disaggregated by supply type. Water quality deteriorated substantially between source and stored water. The mean percentage of contaminated samples (noncompliance) at the source was 46% (95% CI: 33, 60%), whereas mean noncompliance in HSW was 75% (95% CI: 64, 84%). Water supply type was significantly associated with noncompliance at the source (p < 0.001) and in HSW (p = 0.03). Source water (OR = 0.2; 95% CI: 0.1, 0.5) and HSW (OR = 0.3; 95% CI: 0.2, 0.8) from piped supplies had significantly lower odds of contamination compared with non-piped water, potentially due to residual chlorine. Piped water is less likely to be contaminated compared with other water supply types at both the source and in HSW. A focus on upgrading water services to piped supplies may help improve safety, including for those drinking stored water.

Tracking pathogen transmission at the human-wildlife interface: banded mongoose and Escherichia coli.

PubMed

Pesapane, R; Ponder, M; Alexander, K A

2013-06-01

A primary challenge to managing emerging infectious disease is identifying pathways that allow pathogen transmission at the human-wildlife interface. Using Escherichia coli as a model organism, we evaluated fecal bacterial transmission between banded mongoose (Mungos mungo) and humans in northern Botswana. Fecal samples were collected from banded mongoose living in protected areas (n = 87, 3 troops) and surrounding villages (n = 92, 3 troops). Human fecal waste was collected from the same environment (n = 46). Isolates were evaluated for susceptibility to 10 antibiotics. Resistant E. coli isolates from mongoose were compared to human isolates using rep-PCR fingerprinting and MLST-PCR. Antimicrobial resistant isolates were identified in 57 % of the mongoose fecal samples tested (range 31-78% among troops). At least one individual mongoose fecal sample demonstrated resistance to each tested antibiotic, and multidrug resistance was highest in the protected areas (40.9%). E. coli isolated from mongoose and human sources in this study demonstrated an extremely high degree of genetic similarity on rep-PCR (AMOVA, F ST = 0.0027, p = 0.18) with a similar pattern identified on MLST-PCR. Human waste may be an important source of microbial exposure to wildlife. Evidence of high levels of antimicrobial resistance even within protected areas identifies an emerging health threat and highlights the need for improved waste management in these systems.

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. Copyright © 2016 Elsevier Ltd. All rights reserved.

The influence of land-use composition on fecal contamination of riverine source water in southern British Columbia

NASA Astrophysics Data System (ADS)

St Laurent, Jacques; Mazumder, Asit

2012-12-01

The potential for riverine drinking source water to become contaminated with pathogens is related to the production and transport of fecal waste from within the local catchment area. Identifying specific relationships between land-use types and fecal contamination in riverine water provides an indication of the risk associated with land-use change and helps to target mitigation measures toward land-use types of concern. Fecal coliform (FC) data from 42 riverine sites across British Columbia (BC), Canada, were examined in relation to land-use composition (including 16 land-use types) in the local catchment area. FC concentration significantly increased in relation to anthropogenic land-use impacts but was negatively associated with undisturbed and high-elevation land types. Regression tree analysis identified that highest FC concentrations occurred in catchments characterized by more than 12.5% agricultural land and more than 1.6% urban land. Furthermore, the risk of violation of the BC partial treatment raw drinking water quality guideline for FC concentration (100 CFU 100 mL-1) increased in relation to agricultural impacts. Additional factors, such as sewage treatment discharge, low dilution in smaller streams, and higher temperatures, were associated with higher FC concentration among sites with similar levels of agricultural development. These results identify land-use types that present the greatest threat to riverine contamination, namely agricultural and urban land, and indicate the proportion of such land use associated with high contamination. Land use should be managed and source water protection should be targeted in light of these results so as to minimize the risk of surface water exposure to fecal contaminants.

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.

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

Title: Water Quality Monitoring to Restore and Enhance Lake Herrick

NASA Astrophysics Data System (ADS)

Kannan, A.; Saintil, T.; Radcliffe, D. E.; Rasmussen, T. C.

2017-12-01

Lake Allyn M. Herrick is about 1.5 km2 and covers portions of the University of Georgia's East campus, the Oconee forest, residential and commercial land use. Lake Herrick, a 15-acre water body established in 1982 at the University of Georgia's campus was closed in 2002 for recreation due to fecal contamination, color change, and heavy sedimentation. Subsequent monitoring confirmed cyanobacterium blooms on the surface of lake and nutrient concentration especially phosphorus was one of the primary reasons. However, no studies have been done on lake inflows and outflows after 2005 in terms of nutrients and fecal Indicator bacteria. Two inflow tributaries and the outlet stream were monitored for discharge, E. coli, total coliform, forms of nitrogen and phosphorus and other water quality parameters during base flow and storm conditions. External environmental factors like precipitation, land-use/location, discharge, and internal factors within the water like temperature, DO, pH, conductivity, and turbidity influencing fecal indicator bacteria and nutrients will be discussed with data collected from the inflows/outflow between February 2016 to October 2017. Following this, microbial source tracking methods were also used to detect the bacterial source in the samples specific to a ruminant or human host. The source tracking data will be presented during the timeframe of January 2017 to September 2017, to draw a conclusion on the potential source of fecal contamination. The future aim of the project will include modeling flow and bacteria at the watershed scale in order to make management decisions to restore the lake for recreational uses where green infrastructure could play a key role.

Effect of ingestion of soy yogurt on intestinal parameters of rats fed on a beef-based animal diet

PubMed Central

Bedani, Raquel; Pauly-Silveira, Nadiége Dourado; Cano, Veridiana Soares Pereira; Valentini, Sandro Roberto; de Rossi, Graciela Font; Valdez, Elizeu Antonio

2011-01-01

The aim of this study was to investigate whether the ingestion of soy yogurt fermented with Enterococcus faecium CRL 183 would modify the intestinal count of enterococci, fecal pH and ammonia content in rats fed on a diet containing red meat. The rats were placed in 4 groups: for 60 days, group I was given a standard casein-based rodent feed and groups II-IV, the beef-based feed. From day 30, groups III-IV also received the following products: III) soy yogurt; IV) suspension of E. faecium CRL 183. At the start and on days 30 and 60, feces were collected for the determination of pH, ammonia content, count of enterococci and identification of their species. On day 60, rats were sacrificed and their colons also removed for count of enterococci and identification of their species. Rats that ingested soy yogurt showed no significant change (P<0.05) in fecal counts of Enterococcus spp., but, this rat group showed a higher count of E. faecium than rats that ingested suspension of E. faecium CRL 183. The ingestion of soy yogurt and E. faecium culture caused a significant rise (P < 0.05) in fecal pH and ammonia content. Our results suggest that consumption of soy yogurt fermented with E. faecium CRL 183 and L. helveticus subsp. jugurti could change the species of Enterococcus spp. present in the feces and colon of rats fed on a beef-based diet. However, the fermented soy product and the pure culture of E. faecium CRL 183 also induced undesirable effects such as the increase of fecal pH and ammonia content in the feces of rats fed on a beef-based diet. PMID:24031747

Effects of Saccharomyces boulardii on fecal short-chain fatty acids and microflora in patients on long-term total enteral nutrition

PubMed Central

Schneider, Stéphane M; Girard-Pipau, Fernand; Filippi, Jérôme; Hébuterne, Xavier; Moyse, Dominique; Hinojosa, Gustavo Calle; Pompei, Anne; Rampal, Patrick

2005-01-01

AIM: To assess the effects of Sb on fecal flora and short-chain fatty acids (SCFA) in patients on long-term TEN. METHODS: Ten patients (3 females, 7 males, 59±5.5 years), on TEN for a median of 13 mo (1-125), and 15 healthy volunteers (4 females, 11 males, 32±2.0 years) received Sb (0.5 g bid PO) for 6 d. Two stool samples were taken before, on the last 2 d and 9-10 d after treatment, for SCFA measurement and for culture and bacterial identification. Values (mean±SE) were compared using sign tests and ANOVA. RESULTS: Fecal butyrate levels were lower in patients (10.1±2.9 mmol/kg) than in controls (19.2±2.9, P = 0.02). Treatment with Sb increased total fecal SCFA levels in patients (150.2±27.2vs 107.5?8.2 mmol/kg, P = 0.02) but not in controls (129.0±28.6 vs 113.0±15.2 mmol/kg, NS). At the end of treatment with Sb, patients had higher fecal butyrate (16.0±4.4 vs 10.1 [2.9] mmol/kg, P = 0.004). Total SCFAs remained high 9 d after treatment was discon-tinued. Before the treatment, the anaerobe to aerobe ratio was lower in patients compared to controls (2.4±2.3 vs 69.8±1.8, P = 0.003). There were no significant changes in the fecal flora of TEN patients. CONCLUSION: Sb-induced increase of fecal SCFA concentrations (especially butyrate) may explain the preventive effects of this yeast on TEN-induced diarrhea. PMID:16273644

Diagnostic strategies to reveal covert infections with intestinal helminths in dogs.

PubMed

Adolph, Chris; Barnett, Sharon; Beall, Melissa; Drake, Jason; Elsemore, David; Thomas, Jennifer; Little, Susan

2017-11-30

Intestinal helminths are common in dogs in the United States, particularly non-treated dogs in animal shelters, but surveys by fecal flotation may underestimate their prevalence. To determine the prevalence of intestinal helminths and evaluate the ability of fecal flotation and detection of nematode antigen to identify those infections, contents of the entire gastrointestinal tract of 97 adult (>1year) dogs previously identified for humane euthanasia at two animal control shelters in northeastern Oklahoma, USA, were screened. All helminths recovered were washed in saline and fixed prior to enumeration and identification to genus and species. Fecal samples from each dog were examined by passive sodium nitrate (SG 1.33) and centrifugal sugar solution (SG 1.25) flotation. Fecal antigen detection assays were used to confirm the presence of nematode antigen in frozen fecal samples from 92 dogs. Necropsy examination revealed Ancylostoma caninum in 45/97 (46.4%), Toxocara canis in 11/97 (11.3%), Trichuris vulpis in 38/97 (39.2%), Dipylidium caninum in 48/97 (49.5%), and Taenia sp. in 7/97 (7.2%) dogs. Passive fecal flotation identified 38/45 (84.4%) A. caninum, 6/11 (54.5%) T. canis, 26/38 (68.4%) T. vulpis, 2/48 (4.2%) D. caninum, and 1/7 (14.3%) Taenia sp. infections, while centrifugal flotation combined with antigen detection assays identified A. caninum in 97.7% (43/44), T. canis in 77.8% (7/9), and T. vulpis in 83.3% (30/36) of infected dogs based on necropsy recovery of nematodes. Taken together, these data indicate that detection of nematode antigen is a useful adjunct to microscopic examination of fecal samples for parasite eggs, and that this approach can improve diagnostic sensitivity for intestinal nematode infections in dogs. Copyright © 2017 Elsevier B.V. All rights reserved.

Identification of Bacterial DNA Markers for the Detection of Human and Cattle Fecal Pollution - SLIDES

EPA Science Inventory

Technological advances in DNA sequencing and computational biology allow scientists to compare entire microbial genomes. However, the use of these approaches to discern key genomic differences between natural microbial communities remains prohibitively expensive for most laborato...

IDENTIFICATION OF BACTERIAL DNA MARKERS FOR THE DETECTION OF HUMAN AND CATTLE FECAL POLLUTION

EPA Science Inventory

Technological advances in DNA sequencing and computational biology allow scientists to compare entire microbial genomes. However, the use of these approaches to discern key genomic differences between natural microbial communities remains prohibitively expensive for most laborato...

Microbial source tracking in shellfish harvesting waters in the Gulf of Nicoya, Costa Rica.

PubMed

Symonds, E M; Young, S; Verbyla, M E; McQuaig-Ulrich, S M; Ross, E; Jiménez, J A; Harwood, V J; Breitbart, M

2017-03-15

Current microbial water quality monitoring is generally limited to culture-based measurements of fecal indicator bacteria (FIB). Given the many possible sources of fecal pollution within a watershed and extra-intestinal FIB reservoirs, it is important to determine source(s) of fecal pollution as a means to improve water quality and protect public health. The principal objective of this investigation was to characterize the microbial water quality of shellfish harvesting areas in the Gulf of Nicoya, Costa Rica during 2015. In order to achieve this objective, the specificity and sensitivity of 11 existing microbial source tracking (MST) PCR assays, associated with cows (BacCow), dogs (BacCan, DogBac), domestic wastewater (PMMoV), general avian (GFD), gulls (Gull2), horses (HorseBac, HoF), humans (HF183, HPyV), and pigs (PF), were evaluated using domestic wastewater and animal fecal samples collected from the region. The sensitivity of animal-associated assays ranged from 13 to 100%, while assay specificity ranged from 38 to 100%. The specificity of pepper mild mottle virus (PMMoV) and human polyomavirus (HPyV) was 100% for domestic wastewater, as compared to 94% specificity of the HF183 Bacteroidales marker. PMMoV was identified as a useful domestic wastewater-associated marker, with concentrations as high as 1.1 × 10 5 copies/ml and 100% sensitivity and specificity. Monthly surface water samples collected from four shellfish harvesting areas were analyzed using culture-based methods for Escherichia coli as well as molecular methods for FIB and a suite of MST markers, which were selected for their specificity in the region. While culturable E. coli results suggested possible fecal pollution during the monitoring period, the absence of human/domestic wastewater-associated markers and low FIB concentrations determined using molecular methods indicated sufficient microbial water quality for shellfish harvesting. This is the first study to our knowledge to test the performance of MST markers in Costa Rica as well as in Central America. Given the lack of wastewater treatment and the presence of secondary sources of FIB, this study highlights the importance of an MST toolbox approach to characterize water quality in tropical regions. Furthermore, it confirms and extends the geographic range of PMMoV as an effective tool for monitoring domestic wastewater pollution. Copyright © 2017 Elsevier Ltd. All rights reserved.

40 CFR 141.202 - Tier 1 Public Notice-Form, manner, and frequency of notice.

Code of Federal Regulations, 2010 CFR

2010-07-01

... fecal coliform or E. coli are present in the water distribution system (as specified in § 141.63(b)), or when the water system fails to test for fecal coliforms or E. coli when any repeat sample tests...) Detection of E. coli, enterococci, or coliphage in source water samples as specified in § 141.402(a) and...

40 CFR 141.202 - Tier 1 Public Notice-Form, manner, and frequency of notice.

Code of Federal Regulations, 2011 CFR

2011-07-01

... fecal coliform or E. coli are present in the water distribution system (as specified in § 141.63(b)), or when the water system fails to test for fecal coliforms or E. coli when any repeat sample tests...) Detection of E. coli, enterococci, or coliphage in source water samples as specified in § 141.402(a) and...

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...

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...

Molecular Investigations of Bacteroides as Microbial Source Tracking Tools in Southeast Louisiana Watersheds

NASA Astrophysics Data System (ADS)

Schulz, C. J.; Childers, G. W.; Engel, A. S.

2006-12-01

Microbial Source Tracking (MST) is a developing field that is gaining increased attention. MST refers to a host of techniques that discriminates among the origins of fecal material found in natural waters from different sources (e.g. human, livestock, and wildlife) by using microbial indicator species with specificity to only certain host organisms. The development of species-specific molecular markers would allow for better evaluation of public health risks and tracking of nutrient sources impacting a watershed. Although several MST methods have been reported with varying levels of success, few offer general applicability for natural waters due to spatial and temporal constraints associated with these methods. One group of molecular MST markers that show promise for broad environmental applications are molecular 16S rDNA probes for Bacteroides. This method is based on 16S rDNA detection directly from environmental samples without the need for a preliminary cultivation step. In this study we have expanded previous sampling efforts to compile a database of over 1000 partial 16S rRNA Bacteroides genes retrieved from the fecal material of 15 different host species (human, cat, dog, pig, kangaroo). To characterize survival of Bacteroides outside of the host, survival time of the Bacteroides marker was compared to that of E.coli under varying natural environmental conditions (temperature and salinity). Bacteroides displayed a survival curve with shouldering and tailing similar to that of E.coli, but log reduction times differed with treatment. In summary, MST marker stability was identified within host species and the overall Bacteroides community structure correlated to host diet, suggesting that detection of a Bacteroides community could confidently identify fecal contamination point sources. Natural water samples from southeast Louisiana were collected for MST including the Tangipahoa River watershed where the source of fecal contamination has been hotly debated. The Bacteroides tool repeatedly demonstrated the presence of cattle related Bacteroides markers and the absence of human markers.This study is now being expanded to include the entire Lake Pontchartrain Basin.

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. 

Molecular Determination of Fasciola Spp. Isolates from Domestic Ruminants Fecal Samples in the Northwest of Iran

PubMed Central

IMANI BARAN, Abbas; CHERAGHI SARAY, Habib; KATIRAEE, Farzad

2017-01-01

Background: Fasciola species are the main causes for fascioliasis with great financial losses and are among the most important food/water-borne parasites worldwide. The basic proceedings such as epidemiology and effective control of fascioliasis rely mainly on precise identification of Fasciola species. The present study was conducted to determine the Fasciola species in ruminant fecal samples from East Azerbaijan Province in Iran. Methods: Overall, 2012 fecal samples were collected and processed initially for microscopic examination of Fasciola eggs in 2014–15. Then, recovered eggs were subjected to molecular identification. A fragment of 618 bp of the 28S rRNA gene pertaining to Fasciola genus was amplified under PCR. The amplified fragment was restricted by fast digest Ava II enzyme in order to a Restriction Fragment Length Polymorphism. Results: Based on microscopic examination, 72 samples were infected, from which, 10 and 62 cases pertained to cattle and sheep samples respectively. Based on RFLP, the PCR products restricted by the Ava II restriction enzyme produced 529 bp fragments only. According to the positive controls, all restriction patterns were related to Fasciola hepatica, while no restriction patterns were linked to F. gigantica. Conclusion: Based on PCR-RFLP, F. hepatica was dominant species in animals of the studied areas and no evidence of F. gigantica was observed. Therefore, further field studies to verify these results are suggested. PMID:28761485

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.

Cloning and characterization of a novel α-amylase from a fecal microbial metagenome.

PubMed

Xu, Bo; Yang, Fuya; Xiong, Caiyun; Li, Junjun; Tang, Xianghua; Zhou, Junpei; Xie, Zhenrong; Ding, Junmei; Yang, Yunjuan; Huang, Zunxi

2014-04-01

To isolate novel and useful microbial enzymes from uncultured gastrointestinal microorganisms, a fecal microbial metagenomic library of the pygmy loris was constructed. The library was screened for amylolytic activity, and 8 of 50,000 recombinant clones showed amylolytic activity. Subcloning and sequence analysis of a positive clone led to the identification a novel gene (amyPL) coding for α-amylase. AmyPL was expressed in Escherichia coli BL21 (DE3) and the purified AmyPL was enzymatically characterized. This study is the first to report the molecular and biochemical characterization of a novel α-amylase from a gastrointestinal metagenomic library.

MICROBIAL SOURCE TRACKING: DIFFERENT USES AND APPROACHES

EPA Science Inventory

Microbial Source Tracking (MST) methods are used to determine the origin of fecal pollution impacting natural water systems. Several methods require the isolation of pure cultures in order to develop phenotypic or genotypic fingerprint libraries of both source and water bacterial...

Real-time PCR evaluation of Strongylus vulgaris in horses on farms in Denmark and Central Kentucky.

PubMed

Nielsen, M K; Olsen, S N; Lyons, E T; Monrad, J; Thamsborg, S M

2012-12-21

Strongyle parasites are ubiquitous in grazing horses, and the large strongyle Strongylus vulgaris is considered the most pathogenic helminth parasite of horses. Recent investigations have suggested an association between occurrence of this parasite and usage of selective therapy based on regular fecal egg counts. The established diagnostic method for S. vulgaris involves larval culture and subsequent morphological identification of third stage larvae under the microscope. Recently, a real-time PCR assay was developed and validated for the detection and semi-quantification of S. vulgaris eggs in equine fecal samples. The purposes of the present study were (a) to determine the presence of S. vulgaris by real-time PCR in Danish and American horses on farms using vastly different anthelmintic treatment regimens and (b) to evaluate the association between larval culture results and the PCR. A total of 991 horses representing 53 different horse farms in Denmark and Central Kentucky were studied. Fresh fecal samples were collected from all horses, and strongyle eggs retrieved for DNA extraction and subsequent real-time PCR analysis. Individual larval cultures were performed on the Danish part of the data set (663 horses on 42 farms). On the Danish farms, the S. vulgaris PCR prevalence was found to be 9.2% on farms not basing parasite control on fecal egg counts, and 14.1% on farms using selective therapy. No horses were PCR positive in the American part of the study (328 horses on 11 farms). Kappa-values indicated a moderate agreement between PCR and larval culture results, while McNemar tests revealed no statistical difference between the paired proportions. Significant associations were found between PCR cycle of threshold (Ct) value groups and larval culture counts. Results indicate that both diagnostic methods can be useful for determining the occurrence of S. vulgaris on horse farms, but that they both are affected by potential sources of error. The PCR results confirmed previous findings suggesting that S. vulgaris can reemerge under selective therapy regimens. Copyright © 2012 Elsevier B.V. All rights reserved.

Molecular identification of coliform bacteria isolated from drinking water reservoirs with traditional methods and the Colilert-18 system.

PubMed

Kämpfer, Peter; Nienhüser, Anita; Packroff, Gabriele; Wernicke, Frank; Mehling, Arnd; Nixdorf, Katja; Fiedler, Stefanie; Kolauch, Claudia; Esser, Michael

2008-07-01

The accuracy of a traditional method (lactose utilization with acid and gas production) for the detection of coliform bacteria and E. coli was tested in comparison with method ISO 9308-1 (based on acid formation from lactose) and the Colilert-18 system (detection of beta-galactosidase). A total of 345 isolates were identified after isolation from water samples using API 20E strips. The Colilert-18 led to the highest number of positive findings (95% of the isolates were assigned to coliforms), whereas the ISO-9308-1 method resulted only in 29% coliform findings. With the traditional method only 15% were rated positive. Most of the isolates were identified by the API 20E system as Enterobacter spp. (species of the Enterobacter cloacae complex), Serratia spp., Citrobacter spp.and Klebsiella spp.; but species identification remained vague in several cases. A more detailed identification of 126 pure cultures by using 16S rRNA gene sequence analysis and analysis of the hsp60 gene resulted in the identification of Enterobacter nimipressuralis, E. amnigenus, E. asburiae, E. hormaechei, and Serratia fonticola as predominat coliforms. These species are beta-galactosidase positive, but show acid formation from lactose often after a prolonged incubation time. They are often not of fecal origin and may interfere with the ability to accurately detect coliforms of fecal origin.

Towards the Fecal Metabolome Derived from Moderate Red Wine Intake

PubMed Central

Jiménez-Girón, Ana; Muñoz-González, Irene; Martín-Álvarez, Pedro J.; Moreno-Arribas, María Victoria; Bartolomé, Begoña

2014-01-01

Dietary polyphenols, including red wine phenolic compounds, are extensively metabolized during their passage through the gastrointestinal tract; and their biological effects at the gut level (i.e., anti-inflammatory activity, microbiota modulation, interaction with cells, among others) seem to be due more to their microbial-derived metabolites rather than to the original forms found in food. In an effort to improve our understanding of the biological effects that phenolic compounds exert at the gut level, this paper summarizes the changes observed in the human fecal metabolome after an intervention study consisting of a daily consumption of 250 mL of wine during four weeks by healthy volunteers (n = 33). It assembles data from two analytical approaches: (1) UPLC-ESI-MS/MS analysis of phenolic metabolites in fecal solutions (targeted analysis); and (2) UHPLC-TOF MS analysis of the fecal solutions (non-targeted analysis). Both approaches revealed statistically-significant changes in the concentration of several metabolites as a consequence of the wine intake. Similarity and complementarity between targeted and non-targeted approaches in the analysis of the fecal metabolome are discussed. Both strategies allowed the definition of a complex metabolic profile derived from wine intake. Likewise, the identification of endogenous markers could lead to new hypotheses to unravel the relationship between moderate wine consumption and the metabolic functionality of gut microbiota. PMID:25532710

Effects of added chelated trace minerals, organic selenium, yeast culture, direct-fed microbials, and Yucca schidigera extract in horses: II. Nutrient excretion and potential environmental impact.

PubMed

Gordon, M E; Edwards, M S; Sweeney, C R; Jerina, M L

2013-08-01

The objective of this study was to test the hypothesis that an equine diet formulated with chelated trace minerals, organic selenium, yeast culture, direct-fed microbials (DFM) and Yucca schidigera extract would decrease excretion of nutrients that have potential for environmental impact. Horses were acclimated to 100% pelleted diets formulated with (ADD) and without (CTRL) the aforementioned additives. Chelated sources of Cu, Zn, Mn, and Co were included in the ADD diet at a 100% replacement rate of sulfate forms used in the CTRL diet. Additionally, the ADD diet included organic selenium yeast, DFM, and Yucca schidigera extract. Ten horses were fed the 2 experimental diets during two 42-d periods in a crossover design. Total fecal and urine collection occurred during the last 14 d of each period. Results indicate no significant differences between Cu, Zn, Mn, and Co concentrations excreted via urine (P > 0.05) due to dietary treatment. There was no difference between fecal Cu and Mn concentrations (P > 0.05) based on diet consumed. Mean fecal Zn and Co concentrations excreted by horses consuming ADD were greater than CTRL (P < 0.003). Differences due to diet were found for selenium fecal (P < 0.0001) and urine (P < 0.0001) excretions, with decreased concentrations found for horses consuming organic selenium yeast (ADD). In contrast, fecal K (%) was greater (P = 0.0421) for horses consuming ADD, whereas concentrations of fecal solids, total N, ammonia N, P, total ammonia, and fecal output did not differ between dietary treatments (P > 0.05). In feces stockpiled to simulate a crude composting method, no differences (P > 0.05) due to diet were detected for particle size, temperature, moisture, OM, total N, P, phosphate, K, moisture, potash, or ammonia N (P > 0.05). Although no difference (P = 0.2737) in feces stockpile temperature due to diet was found, temperature differences over time were documented (P < 0.0001). In conclusion, the addition of certain chelated mineral sources, organic Se yeast, DFM, and Yucca schidigera extract did not decrease most nutrient concentrations excreted. Horses consuming organic selenium as part of the additive diet had lower fecal and urine Se concentrations, as well as greater fecal K concentrations.

Long-term monitoring of molecular markers can distinguish different seasonal patterns of fecal indicating bacteria sources.

PubMed

Riedel, Timothy E; Thulsiraj, Vanessa; Zimmer-Faust, Amity G; Dagit, Rosi; Krug, Jenna; Hanley, Kaitlyn T; Adamek, Krista; Ebentier, Darcy L; Torres, Robert; Cobian, Uriel; Peterson, Sophie; Jay, Jennifer A

2015-03-15

Elevated levels of fecal indicator bacteria (FIB) have been observed at Topanga Beach, CA, USA. To identify the FIB sources, a microbial source tracking study using a dog-, a gull- and two human-associated molecular markers was conducted at 10 sites over 21 months. Historical data suggest that episodic discharge from the lagoon at the mouth of Topanga Creek is the main source of bacteria to the beach. A decline in creek FIB/markers downstream from upper watershed development and a sharp increase in FIB/markers at the lagoon sites suggest sources are local to the lagoon. At the lagoon and beach, human markers are detected sporadically, dog marker peaks in abundance mid-winter, and gull marker is chronically elevated. Varied seasonal patterns of FIB and source markers were identified showing the importance of applying a suite of markers over long-term spatial and temporal sampling to identify a complex combination of sources of contamination. Copyright © 2014 Elsevier Ltd. All rights reserved.

Coliform contamination of a coastal embayment: Sources and transport pathways

USGS Publications Warehouse

Weiskel, P.K.; Howes, B.L.; Heufelder, G.R.

1996-01-01

Fecal bacterial contamination of nearshore waters has direct economic impacts to coastal communities through the loss of shellfisheries and restrictions of recreational uses. We conducted seasonal measurements of fecal coliform (FC) sources and transport pathways contributing to FC contamination of Buttermilk Bay, a shallow embayment adjacent to Buzzards Bay, MA. Typical of most coastal embayments, there were no direct sewage discharges (i.e., outfalls), and fecal bacteria from human, domestic animal, and wildlife pools entered open waters primarily through direct deposition or after transport through surface waters or groundwaters. Direct fecal coliform inputs to bay waters occurred primarily in winter (December-March) from waterfowl, ~33 x 1012 FC yr-1 or ~67% of the total annual loading. Effects of waterfowl inputs on bay FC densities were mitigated by their seasonality, wide distribution across the bay surface, and the apparent limited dispersal from fecal pellets. On-site disposal of sewage by septic systems was the single largest FC source in the watershed-embayment system, 460 x 1012 FC yr-1, but due to attenuation during subsurface transport only a minute fraction, < 0.006 x 1012 FC yr-1, reached bay waters (<0.01% of annual input to bay). Instead, surface water flows, via storm drains and natural streams under both wet- and dry-weather conditions, contributed the major terrestrial input, 12 x 1012 FC yr-1 (24% of annual input), all from animal sources. Since most of the surface water FC inputs were associated with periodic, short-duration rain events with discharge concentrated in nearshore zones, wet-weather flows were found to have a disproportionately high impact on nearshore FC levels. Elution of FC from shoreline deposits of decaying vegetation (wrack) comprised an additional coliform source. Both laboratory and field experiments suggest significant elution of bacteria from wrack, ~3 x 1012 FC yr-1 on a bay-wide basis (6% of annual input), primarily by periodic tidal flooding and possibly by major rain events. Release of coliforms during resuspension of subtidal sediments was estimated to be a minor source in this system (<1.5 x 1012 FC yr-1 or < 3% of annual input), primarily associated with large storm events in the fall and winter. Based upon the relative source strengths and the spatial and temporal patterns of FC input to Buttermilk Bay, it appears that management practices in similar settings should account for migratory waterfowl, but remediation efforts should focus on the redirection of stormwater runoff through the groundwater transport pathway.

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 rarefaction curve analysis, richness of bacteria diversity in feces decreased as time progressed. Some pathogens such as Campylobacter were detected only at the beginning, meaning they substantially decayed during the course of our study. Overall, this study indicated: (1) sunlight can influence the community structure and (2) after excretion the fecal bacteria diversity can be significantly changed over time. Future studies should therefore use not only the microbial signature of fresh but also moderately aged fecal samples to develop more accurate community profiles for fecal source tracking.

Linking on-farm dairy management practices to storm-flow fecal coliform loading for California coastal watersheds.

PubMed

Lewis, D J; Atwill, E R; Lennox, M S; Hou, L; Karle, B; Tate, K W

2005-08-01

How and where to improve water quality within an agricultural watershed requires data at a spatial scale that corresponds with individual management decision units on an agricultural operation. This is particularly true in the context of water quality regulations, such as Total Maximum Daily Loads (TMDLs), that identify agriculture as one source of non-point source pollution through larger tributary watershed scale and above and below water quality investigations. We have conducted a systems approach study of 10 coastal dairies and ranches to document fecal coliform concentration and loading to surface waters at the management decision unit scale. Water quality samples were collected on a storm event basis from loading units that included: manure management systems; gutters; storm drains; pastures; and corrals and lots. In addition, in-stream samples were collected above and below the dairy facilities and from a control watershed, managed for light grazing and without a dairy facility or human residence and corresponding septic system. Samples were analyzed for fecal coliform concentration by membrane filtration. Instantaneous discharge was measured for each collected sample. Storm runoff was also calculated using the curve number method (SCS, 1985). Results for a representative dairy as well as the entire 10 dairy data set are presented. Fecal coliform concentrations demonstrate high variability both within and between loading units. Fecal coliform concentrations for pastures range from 206 to 2,288,888 cfu/100 ml and for lots from 1,933 to 166,105,000 cfu/100 ml. Mean concentrations for pastures and lots are 121,298 (SE = 62,222) and 3,155,584 (SE = 1,902,713) cfu/100 ml, respectively. Fecal coliform load from units of concentrated animals and manure are significantly more than units such as pastures while storm flow amounts were significantly less. Compared with results from earlier tributary scale studies in the watershed, this systems approach has generated water quality data that is beneficial for management decisions because of its scale and representation of current management activities. These results are facilitating on-farm changes through the cooperative efforts of dairy managers, regulatory agency staff, and sources of technical and financial assistance.

The current state of knowledge on the interaction of Escherichia coli within vegetative filter strips as a sustainable best management practice to reduce fecal pathogen loading into surface waters

PubMed Central

Olilo, Casianes Owino; Muia, Anastasia Wairimu; Moturi, Wilkister Nyaora; Onyando, Japhet Ogalo; Amber, Ford Roegner

2016-01-01

Agro-pastoral operations have the potential to threaten public health with loading of diverse pathogens into surface waters through overland flow; increasing awareness of the limitations of fecal indicators has led to development of a number of advancements in detection, source tracking and predictive modeling of public health risk. These tools and techniques are beginning to be integrated into management strategies. The objective of this review was to determine the status of current knowledge and challenges of the fate and transport of Escherichia coli in overland flow and their interaction within vegetative filter strip (VFS) as one of these implemented best management practices and to critically evaluate its use in that setting as an indicator organism. With few studies directly focusing on VFS removal of E. coli from overland flow, we critically evaluated the available data on movement of E. coil from fecal source loading to retention and decay or re-release for potential contamination of water ways and pointed out potential limitations in both pathogen-specific removal and its use as an indicator organisms within overland flow and VFS. Critical areas of focus for future studies to reduce gaps in knowledge were identified, and the integration of newer approaches in source tracking, alternative indicators and the use of non-pathogenic surrogates for field testing of existing VFS models was encouraged. With VFS as a growing field of interest as an economical conservation practice and as an avenue for conservation of resources for small-scale agro-pastoral operations, management strategies to reduce initial fecal load from either applied manure constituents or shedding from free-range animals will continue to test the limits in the applications of models to overland flow and VFS management strategies. Further studies at the microscale in understanding discrepancies between low and high pathogenicity strains of E. coil and between E. coil and other fecal pathogens in the context of VFS will be critical. However, nuanced studies are needed to understand either biological or environmental differences in the fate and transport of the diverse types of fecal pathogens within these settings PMID:28042601

Fiber fermentability effects on energy and macronutrient digestibility, fecal traits, postprandial metabolite responses, and colon histology of overweight cats.

PubMed

Fischer, M M; Kessler, A M; de Sá, L R M; Vasconcellos, R S; Filho, F O Roberti; Nogueira, S P; Oliveira, M C C; Carciofi, A C

2012-07-01

Considering the different potential benefits of divergent fiber ingredients, the effect of 3 fiber sources on energy and macronutrient digestibility, fermentation product formation, postprandial metabolite responses, and colon histology of overweight cats (Felis catus) fed kibble diets was compared. Twenty-four healthy adult cats were assigned in a complete randomized block design to 2 groups of 12 animals, and 3 animals from each group were fed 1 of 4 of the following kibble diets: control (CO; 11.5% dietary fiber), beet pulp (BP; 26% dietary fiber), wheat bran (WB; 24% dietary fiber), and sugarcane fiber (SF; 28% dietary fiber). Digestibility was measured by the total collection of feces. After 16 d of diet adaptation and an overnight period without food, blood glucose, cholesterol, and triglyceride postprandial responses were evaluated for 16 h after continued exposure to food. On d 20, colon biopsies of the cats were collected under general anesthesia. Fiber addition reduced food energy and nutrient digestibility. Of all the fiber sources, SF had the least dietary fiber digestibility (P < 0.05), causing the largest reduction of dietary energy digestibility (P < 0.05). The greater fermentability of BP resulted in reduced fecal DM and pH, greater fecal production [g/(cat × d); as-is], and greater fecal concentration of acetate, propionate, and lactate (P < 0.05). For most fecal variables, WB was intermediate between BP and SF, and SF was similar to the control diet except for an increased fecal DM and firmer feces production for the SF diet (P < 0.05). Postprandial evaluations indicated reduced mean glucose concentration and area under the glucose curve in cats fed the SF diet (P < 0.05). Colon mucosa thickness, crypt area, lamina propria area, goblet cell area, crypt mean size, and crypt in bifurcation did not vary among the diets. According to the fiber solubility and fermentation rates, fiber sources can induce different physiological responses in cats, reduce energy digestibility, and favor glucose metabolism (SF), or improve gut health (BP).

Source tracking of leaky sewers: a novel approach combining fecal indicators in water and sediments.

PubMed

Guérineau, Hélène; Dorner, Sarah; Carrière, Annie; McQuaid, Natasha; Sauvé, Sébastien; Aboulfadl, Khadija; Hajj-Mohamad, Mariam; Prévost, Michèle

2014-07-01

In highly urbanized areas, surface water and groundwater are particularly vulnerable to sewer exfiltration. In this study, as an alternative to Microbial Source Tracking (MST) methods, we propose a new method combining microbial and chemical fecal indicators (Escherichia coli (E. coli)) and wastewater micropollutants (WWMPs) analysis both in water and sediment samples and under different meteorological conditions. To illustrate the use of this method, wastewater exfiltration and subsequent infiltration were identified and quantified by a three-year field study in an urban canal. The gradients of concentrations observed suggest that several sources of fecal contamination of varying intensity may be present along the canal, including feces from resident animal populations, contaminated surface run-off along the banks and under bridge crossings, release from contaminated banks, entrainment of contaminated sediments, and most importantly sewage exfiltration. Calculated exfiltration-infiltration volumes varied between 0.6 and 15.7 m(3)/d per kilometer during dry weather, and between 1.1 and 19.5 m(3)/d per kilometer during wet weather. WWMPs were mainly diluted and degraded below detection limits in water. E. coli remains the best exfiltration indicator given a large volume of dilution and a high abundance in the wastewater source. WWMPs are effective for detecting cumulated contamination in sediments from a small volume source and are particularly important because E. coli on its own does not allow source tracking. Copyright © 2014 Elsevier Ltd. All rights reserved.

BACTERIA SOURCE TRACKING AND 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 pollu...

Impacts of Hurricanes Katrina and Rita on the microbial landscape of the New Orleans area

PubMed Central

Sinigalliano, C. D.; Gidley, M. L.; Shibata, T.; Whitman, D.; Dixon, T. H.; Laws, E.; Hou, A.; Bachoon, D.; Brand, L.; Amaral-Zettler, L.; Gast, R. J.; Steward, G. F.; Nigro, O. D.; Fujioka, R.; Betancourt, W. Q.; Vithanage, G.; Mathews, J.; Fleming, L. E.; Solo-Gabriele, H. M.

2007-01-01

Floodwaters in New Orleans from Hurricanes Katrina and Rita were observed to contain high levels of fecal indicator bacteria and microbial pathogens, generating concern about long-term impacts of these floodwaters on the sediment and water quality of the New Orleans area and Lake Pontchartrain. We show here that fecal indicator microbe concentrations in offshore waters from Lake Pontchartrain returned to prehurricane concentrations within 2 months of the flooding induced by these hurricanes. Vibrio and Legionella species within the lake were more abundant in samples collected shortly after the floodwaters had receded compared with samples taken within the subsequent 3 months; no evidence of a long-term hurricane-induced algal bloom was observed. Giardia and Cryptosporidium were detected in canal waters. Elevated levels of fecal indicator bacteria observed in sediment could not be solely attributed to impacts from floodwaters, as both flooded and nonflooded areas exhibited elevated levels of fecal indicator bacteria. Evidence from measurements of Bifidobacterium and bacterial diversity analysis suggest that the fecal indicator bacteria observed in the sediment were from human fecal sources. Epidemiologic studies are highly recommended to evaluate the human health effects of the sediments deposited by the floodwaters. PMID:17488814

Fecal Contamination of Drinking-Water in Low- and Middle-Income Countries: A Systematic Review and Meta-Analysis

PubMed Central

Bain, Robert; Cronk, Ryan; Wright, Jim; Yang, Hong; Slaymaker, Tom; Bartram, Jamie

2014-01-01

Background Access to safe drinking-water is a fundamental requirement for good health and is also a human right. Global access to safe drinking-water is monitored by WHO and UNICEF using as an indicator “use of an improved source,” which does not account for water quality measurements. Our objectives were to determine whether water from “improved” sources is less likely to contain fecal contamination than “unimproved” sources and to assess the extent to which contamination varies by source type and setting. Methods and Findings Studies in Chinese, English, French, Portuguese, and Spanish were identified from online databases, including PubMed and Web of Science, and grey literature. Studies in low- and middle-income countries published between 1990 and August 2013 that assessed drinking-water for the presence of Escherichia coli or thermotolerant coliforms (TTC) were included provided they associated results with a particular source type. In total 319 studies were included, reporting on 96,737 water samples. The odds of contamination within a given study were considerably lower for “improved” sources than “unimproved” sources (odds ratio [OR] = 0.15 [0.10–0.21], I2 = 80.3% [72.9–85.6]). However over a quarter of samples from improved sources contained fecal contamination in 38% of 191 studies. Water sources in low-income countries (OR = 2.37 [1.52–3.71]; p<0.001) and rural areas (OR = 2.37 [1.47–3.81] p<0.001) were more likely to be contaminated. Studies rarely reported stored water quality or sanitary risks and few achieved robust random selection. Safety may be overestimated due to infrequent water sampling and deterioration in quality prior to consumption. Conclusion Access to an “improved source” provides a measure of sanitary protection but does not ensure water is free of fecal contamination nor is it consistent between source types or settings. International estimates therefore greatly overstate use of safe drinking-water and do not fully reflect disparities in access. An enhanced monitoring strategy would combine indicators of sanitary protection with measures of water quality. Please see later in the article for the Editors' Summary PMID:24800926

Prevalence of microbiological contaminants in groundwater sources and risk factor assessment in Juba, South Sudan.

PubMed

Engström, Emma; Balfors, Berit; Mörtberg, Ulla; Thunvik, Roger; Gaily, Tarig; Mangold, Mikael

2015-05-15

In low-income regions, drinking water is often derived from groundwater sources, which might spread diarrheal disease if they are microbiologically polluted. This study aimed to investigate the occurrence of fecal contamination in 147 improved groundwater sources in Juba, South Sudan and to assess potential contributing risk factors, based on bivariate statistical analysis. Thermotolerant coliforms (TTCs) were detected in 66% of the investigated sources, including 95 boreholes, breaching the health-based recommendations for drinking water. A significant association (p<0.05) was determined between the presence of TTCs and the depth of cumulative, long-term prior precipitation (both within the previous five days and within the past month). No such link was found to short-term rainfall, the presence of latrines or damages in the borehole apron. However, the risk factor analysis further suggested, to a lesser degree, that the local topography and on-site hygiene were additionally significant. In summary, the analysis indicated that an important contamination mechanism was fecal pollution of the contributing groundwater, which was unlikely due to the presence of latrines; instead, infiltration from contaminated surface water was more probable. The reduction in fecal sources in the environment in Juba is thus recommended, for example, through constructing latrines or designating protection areas near water sources. The study results contribute to the understanding of microbiological contamination of groundwater sources in areas with low incomes and high population densities, tropical climates and weathered basement complex environments, which are common in urban sub-Saharan Africa. Copyright © 2015 Elsevier B.V. All rights reserved.

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 of an area near the confluence of Sluder Branch and Newfound Creek.

DNA BASED MOLECULAR METHODS FOR BACTERIAL SOURCE TRACKING IN WATERSHEDS

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...

THE EPA MICROBIAL SOURCE TRACKING DOCUMENT

EPA Science Inventory

Beach closures or violations of total maximum daily loads of fecal organisms in watersheds frequently generate a need to identify the major sources of contamination or, at least, determine whether the source is human or animal. A few years ago E. coli ribotyping was the only met...

Trichinosis in a dog.

PubMed

Rice, L; Frongillo, M K; Randolph, J F

1990-08-15

Trichinella spiralis infection was identified by direct fecal examination as the cause of gastrointestinal disease in a dog. The source of infection was believed to be a woodchuck. Management included supportive care and benzimidazole treatment. Vomiting, diarrhea, anorexia, and myalgia are the main signs of trichinosis, but routine fecal examination seldom reveals T spiralis in natural infections. Mebendazole is the recommended anthelmintic and should be used to eliminate intestinal larvae and prevent muscle invasion. Although the prevalence of trichinosis is decreasing in swine, wild mammals may still be a potential source for dogs and cats. Nevertheless, because of the nonspecific clinical signs of trichinosis, many cases probably go undiagnosed.

Identification of a novel circular DNA virus in pig feces

USDA-ARS?s Scientific Manuscript database

Metagenomic analysis of fecal samples collected from a swine with diarrhea detected sequences encoding a replicase (Rep) protein typically found in small circular Rep-encoding ssDNA (CRESS-DNA) viruses. The complete 3,062 nucleotide genome was generated and found to encode two bi-directionally trans...

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

Pathogenic bacteria and microbial-source tracking markers in Brandywine Creek Basin, Pennsylvania and Delaware, 2009-10

USGS Publications Warehouse

Duris, Joseph W.; Reif, Andrew G.; Olson, Leif E.; Johnson, Heather E.

2011-01-01

The City of Wilmington, Delaware, is in the downstream part of the Brandywine Creek Basin, on the main stem of Brandywine Creek. Wilmington uses this stream, which drains a mixed-land-use area upstream, for its main drinking-water supply. Because the stream is used for drinking water, Wilmington is in need of information about the occurrence and distribution of specific fecally derived pathogenic bacteria (disease-causing bacteria) and their relations to commonly measured fecal-indicator bacteria (FIB), as well as information regarding the potential sources of the fecal pollution and pathogens in the basin. This study focused on five routinely sampled sites within the basin, one each on the West Branch and the East Branch of Brandywine Creek and at three on the main stem below the confluence of the West and East Branches. These sites were sampled monthly for 1 year. Targeted event samples were collected on two occasions during high flow and two occasions during normal flow. On the basis of this study, high flows in the Brandywine Creek Basin were related to increases in FIB densities, and in the frequency of selected pathogen and source markers, in the West Branch and main stem of Brandywine Creek, but not in the East Branch. Water exceeding the moderate fullbody-contact single-sample recreational water-quality criteria (RWQC) for Escherichia coli (E. coli) was more likely to contain selected markers for pathogenic E. coli (eaeA,stx1, and rfbO157 gene markers) and bovine fecal sources (E. hirae and LTIIa gene markers), whereas samples exceeding the enterococci RWQC were more likely to contain the same pathogenic markers but also were more likely to carry a marker indicative of human source (esp gene marker). On four sample dates, during high flow between October and March, the West Branch was the only observed potential contributor of selected pathogen and bovine source markers to the main stem of Brandywine Creek. Indeed, the stx2 marker, which indicates a highly virulent type of pathogenic E. coli, was found only in the West Branch and main stem at high flow but was not found in the East Branch under similar conditions. However, it must be noted that throughout the entire year of sampling there were occasions, during both high and normal flows, when both the East and West Branches were potential contributors of pathogen and microbial-source tracking markers to the main stem. Therefore, this study indicates that under selected conditions (high flow, October through March), West Branch Brandywine Creek Basin was the most likely source of elevated FIB densities in the main stem. These elevated densities are associated with more frequent detection of selected pathogenic E. coli markers (rfbO157 stx1) and are associated with MST markers of bovine source. However, during other times of the year, both the West Branch and East Branch Basins are acting as potential sources of FIB and fecally derived pathogens.

Microbial Groundwater Quality Status of Hand-Dug Wells and Boreholes in the Dodowa Area of Ghana

PubMed Central

Lutterodt, George; Hoiting, Yvonne; Kamara, Alimamy K.; Oduro-Kwarteng, Sampson; Foppen, Jan Willem A.

2018-01-01

To assess the suitability of water sources for drinking purposes, samples were taken from groundwater sources (boreholes and hand-dug wells) used for drinking water in the Dodowa area of Ghana. The samples were analyzed for the presence of fecal indicator bacteria (Escherichia coli) and viruses (Adenovirus and Rotavirus), using membrane filtration with plating and glass wool filtration with quantitative polymerase chain reaction (PCR), respectively. In addition, sanitary inspection of surroundings of the sources was conducted to identify their vulnerability to pollution. The presence of viruses was also assessed in water samples from the Dodowa River. More than 70% of the hand-dug wells were sited within 10 m of nearby sources of contamination. All sources contained E. coli bacteria, and their numbers in samples of water between dug wells and boreholes showed no significant difference (p = 0.48). Quantitative PCR results for Adenovirus indicated 27% and 55% were positive for the boreholes and hand-dug wells, respectively. Samples from all boreholes tested negative for the presence of Rotavirus while 27% of the dug wells were positive for Rotavirus. PCR tests of 20% of groundwater samples were inhibited. Based on these results we concluded that there is systemic microbial and fecal contamination of groundwater in the area. On-site sanitation facilities, e.g., pit latrines and unlined wastewater drains, are likely the most common sources of fecal contamination of groundwater in the area. Water abstracted from groundwater sources needs to be treated before use for consumption purposes. In addition, efforts should be made to delineate protected areas around groundwater abstraction points to minimize contamination from point sources of pollution. PMID:29649111

A Systematic Review and Meta-Analysis of Fecal Contamination and Inadequate Treatment of Packaged Water

PubMed Central

Williams, Ashley R.; Bain, Robert E. S.; Fisher, Michael B.; Cronk, Ryan; Kelly, Emma R.; Bartram, Jamie

2015-01-01

Background Packaged water products provide an increasingly important source of water for consumption. However, recent studies raise concerns over their safety. Objectives To assess the microbial safety of packaged water, examine differences between regions, country incomes, packaged water types, and compare packaged water with other water sources. Methods We performed a systematic review and meta-analysis. Articles published in English, French, Portuguese, Spanish and Turkish, with no date restrictions were identified from online databases and two previous reviews. Studies published before April 2014 that assessed packaged water for the presence of Escherichia coli, thermotolerant or total coliforms were included provided they tested at least ten samples or brands. Results A total of 170 studies were included in the review. The majority of studies did not detect fecal indicator bacteria in packaged water (78/141). Compared to packaged water from upper-middle and high-income countries, packaged water from low and lower-middle-income countries was 4.6 (95% CI: 2.6–8.1) and 13.6 (95% CI: 6.9–26.7) times more likely to contain fecal indicator bacteria and total coliforms, respectively. Compared to all other packaged water types, water from small bottles was less likely to be contaminated with fecal indicator bacteria (OR = 0.32, 95%CI: 0.17–0.58) and total coliforms (OR = 0.10, 95%CI: 0.05, 0.22). Packaged water was less likely to contain fecal indicator bacteria (OR = 0.35, 95%CI: 0.20, 0.62) compared to other water sources used for consumption. Conclusions Policymakers and regulators should recognize the potential benefits of packaged water in providing safer water for consumption at and away from home, especially for those who are otherwise unlikely to gain access to a reliable, safe water supply in the near future. To improve the quality of packaged water products they should be integrated into regulatory and monitoring frameworks. PMID:26505745

A Systematic Review and Meta-Analysis of Fecal Contamination and Inadequate Treatment of Packaged Water.

PubMed

Williams, Ashley R; Bain, Robert E S; Fisher, Michael B; Cronk, Ryan; Kelly, Emma R; Bartram, Jamie

2015-01-01

Packaged water products provide an increasingly important source of water for consumption. However, recent studies raise concerns over their safety. To assess the microbial safety of packaged water, examine differences between regions, country incomes, packaged water types, and compare packaged water with other water sources. We performed a systematic review and meta-analysis. Articles published in English, French, Portuguese, Spanish and Turkish, with no date restrictions were identified from online databases and two previous reviews. Studies published before April 2014 that assessed packaged water for the presence of Escherichia coli, thermotolerant or total coliforms were included provided they tested at least ten samples or brands. A total of 170 studies were included in the review. The majority of studies did not detect fecal indicator bacteria in packaged water (78/141). Compared to packaged water from upper-middle and high-income countries, packaged water from low and lower-middle-income countries was 4.6 (95% CI: 2.6-8.1) and 13.6 (95% CI: 6.9-26.7) times more likely to contain fecal indicator bacteria and total coliforms, respectively. Compared to all other packaged water types, water from small bottles was less likely to be contaminated with fecal indicator bacteria (OR = 0.32, 95%CI: 0.17-0.58) and total coliforms (OR = 0.10, 95%CI: 0.05, 0.22). Packaged water was less likely to contain fecal indicator bacteria (OR = 0.35, 95%CI: 0.20, 0.62) compared to other water sources used for consumption. Policymakers and regulators should recognize the potential benefits of packaged water in providing safer water for consumption at and away from home, especially for those who are otherwise unlikely to gain access to a reliable, safe water supply in the near future. To improve the quality of packaged water products they should be integrated into regulatory and monitoring frameworks.

A Customized DNA Microarray for Microbial Source Tracking in Environmental Systems

EPA Science Inventory

It is estimated that more than 160, 000 miles of rivers and streams in the United States are impaired due to the presence of waterborne pathogens. These pathogens typically originate from human and other animal fecal pollution sources; therefore, a rapid microbial source tracking...

Development of Strain-Specific Primers for Identification of Bifidobacterium bifidum BGN4.

PubMed

Youn, So Youn; Ji, Geun Eog; Han, Yoo Ri; Park, Myeong Soo

2017-05-28

Bifidobacterium bifidum BGN4 (BGN4) has many proven beneficial effects, including antiallergy and anticancer properties. It has been commercialized and used in several probiotic products, and thus strain-specific identification of this strain is very valuable for further strain-dependent physiological study. For this purpose, we developed novel multiplex polymerase chain reaction (PCR) primer sets for strain-specific detection of BGN4 in commercial products and fecal samples of animal models. The primer set was tested on seven strains of B. bifidum and 75 strains of the other Bifidobacterium species. The BGN4-specific regions were derived using megaBLAST against genome sequences of various B. bifidum databases and four sets of primers were designed. As a result, only BGN4 produced four PCR products simultaneously whereas the other strains did not. The PCR detection limit using BGN4-specific primer sets was 2.8 × 10 1 CFU/ml of BGN4. Those primer sets also detected and identified BGN4 in the probiotic products containing BNG4 and fecal samples from a BGN4-fed animal model with high specificity. Our results indicate that the PCR assay from this study is an efficient tool for the simple, rapid, and reliable identification of BGN4, for which probiotic strains are known.

MICROBIAL SOURCE TRACKING

EPA Science Inventory

Fecal contamination of waters used for recreation, drinking water, and aquaculture is an environmental problem and poses significant human health risks. The problem is often difficult to correct because the source of the contamination cannot be determined with certainty. Run-of...

Incidence of bacterial enteropathogens in foods from Mexico.

PubMed Central

Wood, L V; Ferguson, L E; Hogan, P; Thurman, D; Morgan, D R; DuPont, H L; Ericsson, C D

1983-01-01

We examined food consumption patterns of U.S. students temporarily living in Guadalajara, Mexico. Consumption of foods prepared in Mexican homes was associated with an increased risk of acquisition of diarrhea. Foods from commercial sources and private Mexican homes in Guadalajara were subsequently examined for contamination with coliforms, fecal coliforms, and bacterial enteropathogens. For comparison, selected restaurant foods were obtained in Houston, Tex. Food obtained from Mexican homes showed generally higher counts of coliforms and fecal coliforms than those obtained from commercial sources in Mexico and Houston. The foods in Mexico, both from homes and commercial sources, commonly contained Escherichia coli and occasionally enterotoxigenic E. coli. Foods in Houston were not contaminated with E. coli or enterotoxigenic E. coli. Salmonella (17 isolates), Shigella (4 isolates), and Aeromonas hydrophila (1 isolate) were found only in the foods obtained from Mexican homes. Enterotoxigenic non-E. coli Enterobacteriaceae was recovered with approximately equal frequency from all food sources. PMID:6354085

Modeling seasonal variability of fecal coliform in natural surface waters using the modified SWAT

NASA Astrophysics Data System (ADS)

Cho, Kyung Hwa; Pachepsky, Yakov A.; Kim, Minjeong; Pyo, JongCheol; Park, Mi-Hyun; Kim, Young Mo; Kim, Jung-Woo; Kim, Joon Ha

2016-04-01

Fecal coliforms are indicators of pathogens and thereby, understanding of their fate and transport in surface waters is important to protect drinking water sources and public health. We compiled fecal coliform observations from four different sites in the USA and Korea and found a seasonal variability with a significant connection to temperature levels. In all observations, fecal coliform concentrations were relatively higher in summer and lower during the winter season. This could be explained by the seasonal dominance of growth or die-off of bacteria in soil and in-stream. Existing hydrologic models, however, have limitations in simulating the seasonal variability of fecal coliform. Soil and in-stream bacterial modules of the Soil and Water Assessment Tool (SWAT) model are oversimplified in that they exclude simulations of alternating bacterial growth. This study develops a new bacteria subroutine for the SWAT in an attempt to improve its prediction accuracy. We introduced critical temperatures as a parameter to simulate the onset of bacterial growth/die-off and to reproduce the seasonal variability of bacteria. The module developed in this study will improve modeling for environmental management schemes.

Conservation Physiology of an Uncatchable Animal: The North Atlantic Right Whale (Eubalaena glacialis).

PubMed

Hunt, Kathleen E; Rolland, Rosalind M; Kraus, Scott D

2015-10-01

The North Atlantic right whale, Eubalaena glacialis (NARW), a critically endangered species that has been under intensive study for nearly four decades, provides an excellent case study for applying modern methods of conservation physiology to large whales. By combining long-term sighting histories of known individuals with physiological data from newer techniques (e.g., body condition estimated from photographs; endocrine status derived from fecal samples), physiological state and levels of stress can be estimated despite the lack of any method for nonlethal capture of large whales. Since traditional techniques for validating blood assays cannot be used in large whales, assays of fecal hormones have been validated using information on age, sex, and reproductive state derived from an extensive NARW photo-identification catalog. Using this approach, fecal glucocorticoids have been found to vary dramatically with reproductive state. It is therefore essential that glucocorticoid data be interpreted in conjunction with reproductive data. A case study correlating glucocorticoids with chronic noise is presented as an example. Keys to a successful research program for this uncatchable species have included: consistent population monitoring over decades, data-sharing across institutions, an extensive photo-identification catalog that documents individual histories, and consistent efforts at noninvasive collection of samples over years. Future research will require flexibility to adjust to changing distributions of populations. © The Author 2015. Published by Oxford University Press on behalf of the Society for Integrative and Comparative Biology. All rights reserved. For permissions please email: journals.permissions@oup.com.

The impact of ecological variability on the reproductive endocrinology of wild female African elephants.

PubMed

Wittemyer, George; Ganswindt, André; Hodges, Keith

2007-03-01

Non-invasive endocrine methods enable investigation of the relationship between ecological variation and ovarian activity and how this impacts on demographic processes. The underlying physiological factors driving high variation in inter-calving intervals among multi-parous African elephants offer an interesting system for such an investigation. This study investigates the relationship between Normalized Differential Vegetation Index (NDVI), an ecosystem surrogate measure of primary productivity, and fecal progestin concentrations among wild female elephants. Matched fecal samples and behavioral data on reproductive activity were collected from 37 focal individuals during the two-year study. Linear mixed models were used to explore the relationship between fecal 5alpha-pregnane-3-ol-20-one concentrations and the independent variables of NDVI, calf sex, female age, gestation day, and time since last parturition. Among both non-pregnant and pregnant females, fecal 5alpha-pregnane-3-ol-20-one concentrations were significantly correlated with time-specific NDVI indicating a strong relationship between ecological conditions and endocrine activity regulating reproduction. In addition, the age of a female and time since her last parturition impacted hormone concentrations. These results indicate that the identification of an individual's reproductive status from a single hormone sample is possible, but difficult to achieve in practice since numerous independent factors, particularly season, impact fecal hormone concentrations. Regardless of season, however, fecal 5alpha-pregnane-3-ol-20-one concentrations below 1 microg/g were exclusively collected from non-pregnant females, which could be used as a threshold value to identify non-pregnant individuals. Collectively the information generated contributes to a better understanding of environmental regulation of reproductive endocrinology in wild elephant populations, information salient to the management and manipulation of population dynamics in this species.

Human Bacteroides and total coliforms as indicators of recent combined sewer overflows and rain events in urban creeks

USGS Publications Warehouse

McGinnis, Shannon; Spencer, Susan K.; Firnstahl, Aaron; Stokdyk, Joel; Borchardt, Mark A.; McCarthy, David; Murphy, Heather

2018-01-01

Combined sewer overflows (CSOs) are a known source of human fecal pollution and human pathogens in urban water bodies, which may present a significant public health threat. To monitor human fecal contamination in water, bacterial fecal indicator organisms (FIOs) are traditionally used. However, because FIOs are not specific to human sources and do not correlate with human pathogens, alternative fecal indicators detected using qPCR are becoming of interest to policymakers. For this reason, this study measured correlations between the number and duration of CSOs and mm of rainfall, concentrations of traditional FIOs and alternative indicators, and the presence of human pathogens in two urban creeks. Samples were collected May–July 2016 and analyzed for concentrations of FIOs (total coliforms and E. coli) using membrane filtration as well as for three alternative fecal indicators (human Bacteroides HF183 marker, human polyomavirus (HPoV), pepper mild mottle virus (PMMoV)) and nine human pathogens using qPCR. Four of the nine pathogens analyzed were detected at these sites including adenovirus, Enterohemorrhagic E. coli, norovirus, and Salmonella. Among all indicators studied, human Bacteroides and total coliforms were significantly correlated with recent CSO and rainfall events, while E. coli, PMMoV, and HPoV did not show consistent significant correlations. Further, human Bacteroides were a more specific indicator, while total coliforms were a more sensitive indicator of CSO and rainfall events. Results may have implications for the use and interpretation of these indicators in future policy or monitoring programs.

Human Bacteroides and total coliforms as indicators of recent combined sewer overflows and rain events in urban creeks.

PubMed

McGinnis, Shannon; Spencer, Susan; Firnstahl, Aaron; Stokdyk, Joel; Borchardt, Mark; McCarthy, David T; Murphy, Heather M

2018-07-15

Combined sewer overflows (CSOs) are a known source of human fecal pollution and human pathogens in urban water bodies, which may present a significant public health threat. To monitor human fecal contamination in water, bacterial fecal indicator organisms (FIOs) are traditionally used. However, because FIOs are not specific to human sources and do not correlate with human pathogens, alternative fecal indicators detected using qPCR are becoming of interest to policymakers. For this reason, this study measured correlations between the number and duration of CSOs and mm of rainfall, concentrations of traditional FIOs and alternative indicators, and the presence of human pathogens in two urban creeks. Samples were collected May-July 2016 and analyzed for concentrations of FIOs (total coliforms and E. coli) using membrane filtration as well as for three alternative fecal indicators (human Bacteroides HF183 marker, human polyomavirus (HPoV), pepper mild mottle virus (PMMoV)) and nine human pathogens using qPCR. Four of the nine pathogens analyzed were detected at these sites including adenovirus, Enterohemorrhagic E. coli, norovirus, and Salmonella. Among all indicators studied, human Bacteroides and total coliforms were significantly correlated with recent CSO and rainfall events, while E. coli, PMMoV, and HPoV did not show consistent significant correlations. Further, human Bacteroides were a more specific indicator, while total coliforms were a more sensitive indicator of CSO and rainfall events. Results may have implications for the use and interpretation of these indicators in future policy or monitoring programs. Copyright © 2018 Elsevier B.V. All rights reserved.

Chapter 11. Community analysis-based methods

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cao, Y.; Wu, C.H.; Andersen, G.L.

2010-05-01

Microbial communities are each a composite of populations whose presence and relative abundance in water or other environmental samples are a direct manifestation of environmental conditions, including the introduction of microbe-rich fecal material and factors promoting persistence of the microbes therein. As shown by culture-independent methods, different animal-host fecal microbial communities appear distinctive, suggesting that their community profiles can be used to differentiate fecal samples and to potentially reveal the presence of host fecal material in environmental waters. Cross-comparisons of microbial communities from different hosts also reveal relative abundances of genetic groups that can be used to distinguish sources. Inmore » increasing order of their information richness, several community analysis methods hold promise for MST applications: phospholipid fatty acid (PLFA) analysis, denaturing gradient gel electrophoresis (DGGE), terminal restriction fragment length polymorphism (TRFLP), cloning/sequencing, and PhyloChip. Specific case studies involving TRFLP and PhyloChip approaches demonstrate the ability of community-based analyses of contaminated waters to confirm a diagnosis of water quality based on host-specific marker(s). The success of community-based MST for comprehensively confirming fecal sources relies extensively upon using appropriate multivariate statistical approaches. While community-based MST is still under evaluation and development as a primary diagnostic tool, results presented herein demonstrate its promise. Coupled with its inherently comprehensive ability to capture an unprecedented amount of microbiological data that is relevant to water quality, the tools for microbial community analysis are increasingly accessible, and community-based approaches have unparalleled potential for translation into rapid, perhaps real-time, monitoring platforms.« less

Evaluation of Two Library-Independent Microbial Source Tracking Methods To Identify Sources of Fecal Contamination in French Estuaries▿

PubMed Central

Gourmelon, Michèle; Caprais, Marie Paule; Ségura, Raphaël; Le Mennec, Cécile; Lozach, Solen; Piriou, Jean Yves; Rincé, Alain

2007-01-01

In order to identify the origin of the fecal contamination observed in French estuaries, two library-independent microbial source tracking (MST) methods were selected: (i) Bacteroidales host-specific 16S rRNA gene markers and (ii) F-specific RNA bacteriophage genotyping. The specificity of the Bacteroidales markers was evaluated on human and animal (bovine, pig, sheep, and bird) feces. Two human-specific markers (HF183 and HF134), one ruminant-specific marker (CF193′), and one pig-specific marker (PF163) showed a high level of specificity (>90%). However, the data suggest that the proposed ruminant-specific CF128 marker would be better described as an animal marker, as it was observed in all bovine and sheep feces and 96% of pig feces. F RNA bacteriophages were detected in only 21% of individual fecal samples tested, in 60% of pig slurries, but in all sewage samples. Most detected F RNA bacteriophages were from genotypes II and III in sewage samples and from genotypes I and IV in bovine, pig, and bird feces and from pig slurries. Both MST methods were applied to 28 water samples collected from three watersheds at different times. Classification of water samples as subject to human, animal, or mixed fecal contamination was more frequent when using Bacteroidales markers (82.1% of water samples) than by bacteriophage genotyping (50%). The ability to classify a water sample increased with increasing Escherichia coli or enterococcus concentration. For the samples that could be classified by bacteriophage genotyping, 78% agreed with the classification obtained from Bacteroidales markers. PMID:17557850

Differential Decay of Cattle-associated Fecal Indicator ...

EPA Pesticide Factsheets

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 ambient waters. Microbial source tracking (MST) field has evolved in response to a need to identify pollution source(s), but majority of MST markers suffer from the same caveat as FIB, as our understanding of the factors influencing their fate in the environment is limited. Materials: We assessed the effect of water type (freshwater vs marine) and select environmental parameters (indigenous microbiota, ambient sunlight) on decay of FIB and MST markers from cattle manure. Experiments were conducted in situ using a submersible aquatic mesocosm containing dialysis bags filled with mixture of cattle manure and ambient water. Culturable FIB were enumerated by membrane filtration and via qPCR (Entero1a, EC23S) and MST markers were enumerated via qPCR and included general marker of fecal pollution (GenBac3) and cattle-associated subset (Rum2Bac, CowM2, CowM3). Results: Decay of culturable FIB was significantly faster (P > 0.001) than any of the molecular markers irrespective of the water type or experimental conditions. The water type was a significant factor affecting decay (P: 0.008 to < 0.001), although the magnitude of the effect differed among the microbial targets and over time. Presence o

Development of Real-Time PCR to Monitor Groundwater Contaminated by Fecal Sources and Leachate from the Carcass

NASA Astrophysics Data System (ADS)

Park, S.; Kim, H.; Kim, M.; Lee, Y.; Han, J.

2011-12-01

The 2010 outbreak of foot and mouth disease (FMD) in South Korea caused about 4,054 carcass burial sites to dispose the carcasses. Potential environmental impacts by leachate of carcass on groundwater have been issued and it still needs to be studied. Therefore, we tried to develop robust and sensitive tool to immediately determine a groundwater contamination by the leachate from carcass burial. For tracking both an agricultural fecal contamination source and the leachate in groundwater, competitive real-time PCR and PCR method were developed using various PCR primer sets designed to detect E. Coli uidA gene and mtDNA(cytochrome B, cytB) of the animal species such as ovine, porcine, caprine, and bovine. The designed methods were applied to tract the animal species in livestock wastewater and leachate of carcass under appropriate PCR or real-time PCR condition. In the result, mtDNA primer sets for individual (Cow or Pig) and multiple (Cow and Pig) amplification, and E. Coli uidA primers for fecal source amplification were specific and sensitive to target genes. To determine contamination source, concentration of amplified mtDNA and uidA was competitively quantified in Livestock wastewater, leachate of carcass, and groundwater. The highest concentration of mtDNA and uidA showed in leachate of carcass and livestock wastewater, respectively. Groundwater samples possibly contaminated by leachate of carcass were analyzed by this assay and it was able to prove contamination source.

FECAL SOURCE TRACKING BY ANTIBIOTIC RESISTANCE ANALYSIS ON A WATERSHED EXHIBITING LOW RESISTANCE

EPA Science Inventory

The ongoing development of microbial source tracking has made it possible to identify contamination sources with varying accuracy, depending on the method used. The purpose of this study was done to test the efficiency of the antibiotic resistance analysis (ARA) method under low ...

LIBRARY-DEPENDENT MICROBIAL SOURCE TRACKING OF ENTEROCOCCUS SP. USING AFLP AND BOX-PCR

EPA Science Inventory

Library-dependent microbial source tracking (LD MST) methods are one of the approaches used to identify nonpoint sources of fecal contamination in support of total maximum daily load implementation. However, LD MST methods have been questioned due to the high temporal and spatial...

DEVELOPMENT OF HOST-SPECIFIC METAGENOMIC MARKERS FOR MICROBIAL SOURCE TRACKING USING A NOVEL METAGENOMIC APPROACH

EPA Science Inventory

Fecal contamination of source waters is an important issue to the drinking water industry. Improper disposal of animal waste, leaky septic tanks, storm runoff, and wildlife can all be responsible for spreading enteric pathogens into source waters. As a result, methods that can pi...

QUO VADIS SOURCE TRACKING? TOWARDS A STRATEGIC FRAMEWORK FOR ENVIRONMENTAL MONITORING OF FECAL POLLUTION

EPA Science Inventory

Advances in microbial source tracking (MST) have largely been driven by the need to comply with water quality standards based on traditional indicator bacteria. Recently, a number of PCR-based, culture- and library-independent methods have been gaining popularity among source tra...

Toxoplasmosis Update and Public Health Implications

PubMed Central

Fayer, R.

1981-01-01

Toxoplasma gondii has a coccidian life cycle in the intestine of domestic and wild felids that includes a series of asexual and sexual stages and an oocyst stage that is shed in the feces. Oocysts complete their development outside the body, eventually becoming infective for about 350 species of vertebrates including cats and man. The effects of climate on oocyst survival and the physical and biological means of oocyst dispersal are discussed. Infectivity and pathogenicity for livestock species vary. Acute disease results from rapidly multiplying tachyzoites that may be transmitted by carnivorism, transfusion, vertical transmission and other routes. Patent infections may persist for the life of a host as bradyzoites within tissue cysts. Bradyzoites initiate acute infection in other hosts after carnivorism or organ transplantation or in the same host after immunosuppression. Also discussed are: (a) prevalence of T. gondii in livestock as determined by digestion and serological techniques, (b) identification in humans as accomplished by isolation, serological and skin test techniques and (c) identification in cats as accomplished primarily by fecal examinations for oocysts infective for mice. Source of human infections, major outbreaks, treatment, effects on mental health and methods for preventing toxoplasmosis in man and livestock are listed. ImagesFigure 1. PMID:7337909

Investigation into potential transmission sources of Giardia duodenalis in a threatened marsupial (Petrogale penicillata).

PubMed

Vermeulen, Elke T; Ashworth, Deborah L; Eldridge, Mark D B; Power, Michelle L

2015-07-01

Assemblages of the protozoan parasite Giardia duodenalis common in humans and domestic species are increasingly identified in wildlife species, raising concern about the spill-over of pathogens from humans and domestic animals into wildlife. Here, the identity and prevalence of G. duodenalis in populations of a threatened marsupial, the brush-tailed rock-wallaby (Petrogale penicillata), was investigated. Identification of G. duodenalis isolates, across three loci (18S rRNA, β-giardin and gdh), from rock-wallaby fecal samples (n = 318) identified an overall detection rate of 6.3%. No significant difference in G. duodenalis detection was found among captive, wild and supplemented populations. Isolates were assigned to the zoonotic assemblages A and B at 18S rRNA, with sub-assemblages AI and BIV identified at the β-giardin and gdh loci, respectively. Assemblages AI and BIV have previously been identified in human clinical cases, but also in domestic animals and wildlife. The identification of these assemblages in brush-tailed rock-wallabies suggests there are transmission routes of G. duodenalis from humans or other animals to Australian wildlife, both in captivity and in the wild. Copyright © 2015 Elsevier B.V. All rights reserved.

An Analysis of the United States Air Force Nonpoint Source Water Pollution Abatement Program

DTIC Science & Technology

1990-09-01

salts and other minerals. The nmoff frm barnyards and confined feedlots carries organic matter, amcmia, fecal bacteria and other microorganisms, and...nutrients in fertilizers used in lawn care, fecal bacteria from animals (mainly pets and birds), and pesticides are all contained in this runoff...application of pesticides and fertilizers. Less of these products can be applied at optinun intervals and under the proper conditions to reduce the

Hyperspectral imaging technique for detection of poultry fecal residues on food processing equipments

NASA Astrophysics Data System (ADS)

Cho, Byoung-Kwan; Kim, Moon S.; Chen, Yud-Ren

2005-11-01

Emerging concerns about safety and security in current mass production of food products necessitate rapid and reliable inspection for contaminant-free products. Diluted fecal residues on poultry processing plant equipment surface, not easily discernable from water by human eye, are contamination sources for poultry carcasses. Development of sensitive detection methods for fecal residues is essential to ensure safe production of poultry carcasses. Hyperspectral imaging techniques have shown good potential for detecting of the presence of fecal and other biological substances on food and processing equipment surfaces. In this study, use of high spatial resolution hyperspectral reflectance and fluorescence imaging (with UV-A excitation) is presented as a tool for selecting a few multispectral bands to detect diluted fecal and ingesta residues on materials used for manufacturing processing equipment. Reflectance and fluorescence imaging methods were compared for potential detection of a range of diluted fecal residues on the surfaces of processing plant equipment. Results showed that low concentrations of poultry feces and ingesta, diluted up to 1:100 by weight with double distilled water, could be detected using hyperspectral fluorescence images with an accuracy of 97.2%. Spectral bands determined in this study could be used for developing a real-time multispectral inspection device for detection of harmful organic residues on processing plant equipment.

Estimation of Pig Fecal Contamination in a River Catchment by Real-Time PCR Using Two Pig-Specific Bacteroidales 16S rRNA Genetic Markers▿

PubMed Central

Mieszkin, Sophie; Furet, Jean-Pierre; Corthier, Gérard; Gourmelon, Michèle

2009-01-01

The microbiological quality of coastal or river water can be affected by fecal contamination from human or animal sources. To discriminate pig fecal pollution from other pollution, a library-independent microbial source tracking method targeting Bacteroidales host-specific 16S rRNA gene markers by real-time PCR was designed. Two pig-specific Bacteroidales markers (Pig-1-Bac and Pig-2-Bac) were designed using 16S rRNA gene Bacteroidales clone libraries from pig feces and slurry. For these two pig markers, 98 to 100% sensitivity and 100% specificity were obtained when tested by TaqMan real-time PCR. A decrease in the concentrations of Pig-1-Bac and Pig-2-Bac markers was observed throughout the slurry treatment chain. The two newly designed pig-specific Bacteroidales markers, plus the human-specific (HF183) and ruminant-specific (BacR) Bacteroidales markers, were then applied to river water samples (n = 24) representing 14 different sites from the French Daoulas River catchment (Brittany, France). Pig-1-Bac and Pig-2-Bac were quantified in 25% and 62.5%, respectively, of samples collected around pig farms, with concentrations ranging from 3.6 to 4.1 log10 copies per 100 ml of water. They were detected in water samples collected downstream from pig farms but never detected near cattle farms. HF183 was quantified in 90% of water samples collected downstream near Daoulas town, with concentrations ranging between 3.6 and 4.4 log10 copies per 100 ml of water, and BacR in all water samples collected around cattle farms, with concentrations ranging between 4.6 and 6.0 log10 copies per 100 ml of water. The results of this study highlight that pig fecal contamination was not as frequent as human or bovine fecal contamination and that fecal pollution generally came from multiple origins. The two pig-specific Bacteroidales markers can be applied to environmental water samples to detect pig fecal pollution. PMID:19329663

Chemical and biological tracers to determine groundwater flow in karstic aquifer, Yucatan Peninsula

NASA Astrophysics Data System (ADS)

Lenczewski, M.; Leal-Bautista, R. M.; McLain, J. E.

2013-05-01

Little is known about the extent of pollution in groundwater in the Yucatan Peninsula; however current population growth, both from international tourism and Mexican nationals increases the potential for wastewater release of a vast array of contaminants including personal care products, pharmaceuticals (Rx), and pathogenic microorganisms. Pathogens and Rx in groundwater can persist and can be particularly acute in this region where high permeability of the karst bedrock and the lack of top soil permit the rapid transport of contaminants into groundwater aquifers. The objective of this research is to develop and utilize novel biological and chemical source tracking methods to distinguish between different sources of anthropogenic pollution in degraded groundwater. Although several methods have been used successfully to track fecal contamination sources in small scale studies, little is known about their spatial limitations, as source tracking studies rarely include sample collection over a wide geographical area and with different sources of water. In addition, although source tracking methods to distinguish human from animal fecal contamination are widely available, this work has developed source tracking distinguish between separate human populations is highly unique. To achieve this objective, we collected water samples from a series of drinking wells, cenotes (sinkholes), wastewater treatment plants, and injection wells across the Yucatan Peninsula and examine potential source tracers within the collected water samples. The result suggests that groundwater sources impacted by tourist vs. local populations contain different chemical stressors. This work has developed a more detailed understanding of the presence and persistence of personal care products, pharmaceuticals, and fecal indicators in a karstic system; such understanding will be a vital component for the protection Mexican groundwater and human health. Quantification of different pollution sources within groundwater samples identified point sources of pollution, identify potential remediation strategies, and contribute to an improved understanding of the environmental impact of tourism and tourism-generated waste products on this groundwater-dependent ecosystem.

African Non-Human Primates Host Diverse Enteroviruses.

PubMed

Mombo, Illich Manfred; Lukashev, Alexander N; Bleicker, Tobias; Brünink, Sebastian; Berthet, Nicolas; Maganga, Gael D; Durand, Patrick; Arnathau, Céline; Boundenga, Larson; Ngoubangoye, Barthélémy; Boué, Vanina; Liégeois, Florian; Ollomo, Benjamin; Prugnolle, Franck; Drexler, Jan Felix; Drosten, Christian; Renaud, François; Rougeron, Virginie; Leroy, Eric

2017-01-01

Enteroviruses (EVs) belong to the family Picornaviridae and are responsible for mild to severe diseases in mammals including humans and non-human primates (NHP). Simian EVs were first discovered in the 1950s in the Old World Monkeys and recently in wild chimpanzee, gorilla and mandrill in Cameroon. In the present study, we screened by PCR EVs in 600 fecal samples of wild apes and monkeys that were collected at four sites in Gabon. A total of 32 samples were positive for EVs (25 from mandrills, 7 from chimpanzees, none from gorillas). The phylogenetic analysis of VP1 and VP2 genes showed that EVs identified in chimpanzees were members of two human EV species, EV-A and EV-B, and those identified in mandrills were members of the human species EV-B and the simian species EV-J. The identification of two novel enterovirus types, EV-B112 in a chimpanzee and EV-B113 in a mandrill, suggests these NHPs could be potential sources of new EV types. The identification of EV-B107 and EV90 that were previously found in humans indicates cross-species transfers. Also the identification of chimpanzee-derived EV110 in a mandrill demonstrated a wide host range of this EV. Further research of EVs in NHPs would help understanding emergence of new types or variants, and evaluating the real risk of cross-species transmission for humans as well for NHPs populations.

Screening and rapid identification of Campylobacter spp. DNA by FlaA PCR based method on chicken and human fecal samples in Egypt

USDA-ARS?s Scientific Manuscript database

Campylobacter is a foodborne pathogen which has a potential public health concern worldwide. Due to discriminatory problems encountered by conventional isolation of Campylobacter spp. and its genetic similarities, rapid molecular techniques for its genetic characterization are useful. In this study,...

Molecular identification of Enterocytozoon bieneusi, Cryptosporidium, and Giardia in brazilian captive birds

USDA-ARS?s Scientific Manuscript database

A total of 85 fecal samples from captive birds collected from October 2013 to September 2014 in Uberlândia and Belo Horizonte in the state of Minas Gerais (Brazil) were evaluated for the presence of Enterocytozoon bieneusi, Cryptosporidium, and Giardia by PCR. Of these, 3 birds were found positive f...

Development of an Enhanced Metaproteomic Approach for Deepening the Microbiome Characterization of the Human Infant Gut

PubMed Central

2015-01-01

The establishment of early life microbiota in the human infant gut is highly variable and plays a crucial role in host nutrient availability/uptake and maturation of immunity. Although high-performance mass spectrometry (MS)-based metaproteomics is a powerful method for the functional characterization of complex microbial communities, the acquisition of comprehensive metaproteomic information in human fecal samples is inhibited by the presence of abundant human proteins. To alleviate this restriction, we have designed a novel metaproteomic strategy based on double filtering (DF) the raw samples, a method that fractionates microbial from human cells to enhance microbial protein identification and characterization in complex fecal samples from healthy premature infants. This method dramatically improved the overall depth of infant gut proteome measurement, with an increase in the number of identified low-abundance proteins and a greater than 2-fold improvement in microbial protein identification and quantification. This enhancement of proteome measurement depth enabled a more extensive microbiome comparison between infants by not only increasing the confidence of identified microbial functional categories but also revealing previously undetected categories. PMID:25350865

Evaluation of Bovine Feces-Associated Microbial Source Tracking Markers and Their Correlations with Fecal Indicators and Zoonotic Pathogens in a Brisbane, Australia, Reservoir

PubMed Central

Sritharan, T.; Palmer, A.; Sidhu, J. P. S.; Toze, S.

2013-01-01

This study was aimed at evaluating the host specificity and host sensitivity of two bovine feces-associated bacterial (BacCow-UCD and cowM3) and one viral [bovine adenovirus (B-AVs)] microbial source tracking (MST) markers by screening 130 fecal and wastewater samples from 10 target and nontarget host groups in southeast Queensland, Australia. In addition, 36 water samples were collected from a reservoir and tested for the occurrence of all three bovine feces-associated markers along with fecal indicator bacteria (FIB), Campylobacter spp., Escherichia coli O157, and Salmonella spp. The overall host specificity values of the BacCow-UCD, cowM3, and B-AVs markers to differentiate between bovine and other nontarget host groups were 0.66, 0.88, and 1.00, respectively (maximum value of 1.00). The overall host sensitivity values of these markers, however, in composite bovine wastewater and individual bovine fecal DNA samples were 0.93, 0.90, and 0.60, respectively (maximum value of 1.00). Among the 36 water samples tested, 56%, 22%, and 6% samples were PCR positive for the BacCow-UCD, cowM3, and B-AVs markers, respectively. Among the 36 samples tested, 50% and 14% samples were PCR positive for the Campylobacter 16S rRNA and E. coli O157 rfbE genes, respectively. Based on the results, we recommend that multiple bovine feces-associated markers be used if possible for bovine fecal pollution tracking. Nonetheless, the presence of the multiple bovine feces-associated markers along with the presence of potential zoonotic pathogens indicates bovine fecal pollution in the reservoir water samples. Further research is required to understand the decay rates of these markers in relation to FIB and zoonotic pathogens. PMID:23417003

Pepper Mild Mottle Virus as an Indicator of Fecal Pollution ▿

PubMed Central

Rosario, Karyna; Symonds, Erin M.; Sinigalliano, Christopher; Stewart, Jill; Breitbart, Mya

2009-01-01

Accurate indicators of fecal pollution are needed in order to minimize public health risks associated with wastewater contamination in recreational waters. However, the bacterial indicators currently used for monitoring water quality do not correlate with the presence of pathogens. Here we demonstrate that the plant pathogen Pepper mild mottle virus (PMMoV) is widespread and abundant in wastewater from the United States, suggesting the utility of this virus as an indicator of human fecal pollution. Quantitative PCR was used to determine the abundance of PMMoV in raw sewage, treated wastewater, seawater exposed to wastewater, and fecal samples and/or intestinal homogenates from a wide variety of animals. PMMoV was present in all wastewater samples at concentrations greater than 1 million copies per milliliter of raw sewage. Despite the ubiquity of PMMoV in human feces, this virus was not detected in the majority of animal fecal samples tested, with the exception of chicken and seagull samples. PMMoV was detected in four out of six seawater samples collected near point sources of secondary treated wastewater off southeastern Florida, where it co-occurred with several other pathogens and indicators of fecal pollution. Since PMMoV was not found in nonpolluted seawater samples and could be detected in surface seawater for approximately 1 week after its initial introduction, the presence of PMMoV in the marine environment reflects a recent contamination event. Together, these data demonstrate that PMMoV is a promising new indicator of fecal pollution in coastal environments. PMID:19767474

Estimated human health risks from recreational exposures to stormwater runoff containing animal faecal material

EPA Science Inventory

Scientific evidence supporting recreational water quality benchmarks primarily stems from epidemiological studies conducted at beaches impacted by human fecal sources. Epidemiological studies conducted at locations impacted by non-human faecal sources have provided ambiguous and ...

MICROBIAL SOURCE TRACKING GUIDE DOCUMENT

EPA Science Inventory

Approximately 13% of surface waters in the United States do not meet designated use criteria as determined by high densities of fecal indicator bacteria. Although some of the contamination is attributed to point sources such as confined animal feeding operation (CAFO) and wastew...

Outbreak of acute gastroenteritis of unknown etiology caused by contaminated drinking water in a rural village in Austria, August 2006.

PubMed

Meusburger, Stefan; Reichart, Sandra; Kapfer, Sabine; Schableger, Karl; Fretz, Rainer; Allerberger, Franz

2007-01-01

In August 2006 a physician from a rural village reported an outbreak of acute gastroenteritis. An investigation was undertaken in order to determine the magnitude of the outbreak, the source of infection and to prevent further disease. This is the first published outbreak of acute gastroenteritis caused by contaminated drinking water in Austria. For descriptive epidemiology, the investigators had to rely on voluntary cooperation from physicians and patients, data collected by a police officer and data on sick leave reported by physicians to the health insurance system. Microbiological testing of water samples indicated that this cluster was caused by fecal contamination of untreated drinking water. Age and sex distributions were available for 146 of 160 cases: ages ranged from 5 to 91 years (median 45) and 81 cases (55.5%) were female. Stool samples from 14 patients were sent for microbiological analysis: all tested negative for Salmonella, Campylobacter, Shigella and Yersinia enterocolitica. Specimens were not tested for viruses, parasites or enteropathogenic Escherichia coli. In this outbreak no identification was made of pathogenic microorganisms in stool samples from affected patients, despite the occurrence of fecal indicator organisms in samples of drinking water. In outbreaks of gastroenteritis, medical practitioners should encourage microbiological testing beyond the limited routine program. Public health officers must be made aware that the spectrum of routine laboratory tests on stool specimens does not cover the wide array of pathogens capable of causing waterborne outbreaks. The springs serving the affected village originate in a mountainous area of karst formations, and heavy falls of rain that occurred at the beginning of the outbreak may explain introduction of fecal bacteria. In view of the unsolved problem of possible future contamination of springs in karst areas, the water department of this district authority has issued an order requesting installation of a permanent ultraviolet water-treatment facility.

Retrospective Species Identification of Microsporidian Spores in Diarrheic Fecal Samples from Human Immunodeficiency Virus/AIDS Patients by Multiplexed Fluorescence In Situ Hybridization▿

PubMed Central

Graczyk, Thaddeus K.; Johansson, Michael A.; Tamang, Leena; Visvesvara, Govinda S.; Moura, Laci S.; DaSilva, Alexandre J.; Girouard, Autumn S.; Matos, Olga

2007-01-01

In order to assess the applicability of multiplexed fluorescence in situ hybridization (FISH) assay for the clinical setting, we conducted retrospective analysis of 110 formalin-stored diarrheic stool samples from human immunodeficiency virus (HIV)/AIDS patients with intestinal microsporidiosis collected between 1992 and 2003. The multiplexed FISH assay identified microsporidian spores in 94 of 110 (85.5%) samples: 49 (52.1%) were positive for Enterocytozoon bieneusi, 43 (45.8%) were positive for Encephalitozoon intestinalis, 2 (2.1%) were positive for Encephalitozoon hellem, and 9 samples (9.6%) contained both E. bieneusi and E. intestinalis spores. Quantitative spore counts per ml of stool yielded concentration values from 3.5 × 103 to 4.4 × 105 for E. bieneusi (mean, 8.8 × 104/ml), 2.3 × 102 to 7.8 × 104 (mean, 1.5 × 104/ml) for E. intestinalis, and 1.8 × 102 to 3.6 × 102 for E. hellem (mean, 2.7 × 102/ml). Identification of microsporidian spores by multiplex FISH assay was more sensitive than both Chromotrope-2R and CalcoFluor White M2R stains; 85.5% versus 72.7 and 70.9%, respectively. The study demonstrated that microsporidian coinfection in HIV/AIDS patients with intestinal microsporidiosis is not uncommon and that formalin-stored fecal samples older than 10 years may not be suitable for retrospective analysis by techniques targeting rRNA. Multiplexed FISH assay is a reliable, quantitative fluorescence microscopy method for the simultaneous identification of E. bieneusi, E. intestinalis, and E. hellem, as well as Encephalitozoon cuniculi, spores in fecal samples and is a useful tool for assessing spore shedding intensity in intestinal microsporidiosis. The method can be used for epidemiological investigations and applied in clinical settings. PMID:17287331

Global Distribution of Human-Associated Fecal Genetic Markers in Reference Samples from Six Continents.

PubMed

Mayer, René E; Reischer, Georg H; Ixenmaier, Simone K; Derx, Julia; Blaschke, Alfred Paul; Ebdon, James E; Linke, Rita; Egle, Lukas; Ahmed, Warish; Blanch, Anicet R; Byamukama, Denis; Savill, Marion; Mushi, Douglas; Cristóbal, Héctor A; Edge, Thomas A; Schade, Margit A; Aslan, Asli; Brooks, Yolanda M; Sommer, Regina; Masago, Yoshifumi; Sato, Maria I; Taylor, Huw D; Rose, Joan B; Wuertz, Stefan; Shanks, Orin C; Piringer, Harald; Mach, Robert L; Savio, Domenico; Zessner, Matthias; Farnleitner, Andreas H

2018-05-01

Numerous bacterial genetic markers are available for the molecular detection of human sources of fecal pollution in environmental waters. However, widespread application is hindered by a lack of knowledge regarding geographical stability, limiting implementation to a small number of well-characterized regions. This study investigates the geographic distribution of five human-associated genetic markers (HF183/BFDrev, HF183/BacR287, BacHum-UCD, BacH, and Lachno2) in municipal wastewaters (raw and treated) from 29 urban and rural wastewater treatment plants (750-4 400 000 population equivalents) from 13 countries spanning six continents. In addition, genetic markers were tested against 280 human and nonhuman fecal samples from domesticated, agricultural and wild animal sources. Findings revealed that all genetic markers are present in consistently high concentrations in raw (median log 10 7.2-8.0 marker equivalents (ME) 100 mL -1 ) and biologically treated wastewater samples (median log 10 4.6-6.0 ME 100 mL -1 ) regardless of location and population. The false positive rates of the various markers in nonhuman fecal samples ranged from 5% to 47%. Results suggest that several genetic markers have considerable potential for measuring human-associated contamination in polluted environmental waters. This will be helpful in water quality monitoring, pollution modeling and health risk assessment (as demonstrated by QMRAcatch) to guide target-oriented water safety management across the globe.

Effects of feeding pasteurized waste milk to dairy calves on phenotypes and genotypes of antimicrobial resistance in fecal Escherichia coli isolates before and after weaning.

PubMed

Maynou, G; Migura-Garcia, L; Chester-Jones, H; Ziegler, D; Bach, A; Terré, M

2017-10-01

The aim of this study was to evaluate the effects of feeding pasteurized waste milk (pWM) to calves on antimicrobial resistance of fecal Escherichia coli at both phenotypic and genotypic levels. Fifty-two Holstein female calves (3 ± 1.3 d of age) were fed 1 of the 2 different types of milk: milk replacer (MR) without antimicrobials or pWM with β-lactam residues until weaning at 49 d of age. Fecal swabs of all calves were obtained on d 0, 35, and 56 of the study and 3 E. coli isolates per sample were studied. Phenotypic resistance was tested by the disk diffusion method against a panel of 12 antimicrobials. A total of 13 resistance genes consisting of β-lactam, sulfonamide, tetracycline, and aminoglycoside families were examined by PCR. Feeding pWM to calves increased the presence of phenotypic resistance to ampicillin, cephalotin, ceftiofur, and florfenicol in fecal E. coli compared with MR-fed calves. However, the presence of resistance to sulfonamides, tetracyclines, and aminoglycosides was common in dairy calves independent of their milk-feeding source, suggesting other factors apart from the feeding source are involved in the emergence of antimicrobial resistance. Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

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

Cryptosporidium spp. and Giardia spp. in feces and water and the associated exposure factors on dairy farms

PubMed Central

Martins, Felippe Danyel Cardoso; Ferreira, Fernanda Pinto; de Almeida, Jonatas Campos; Ogawa, Liza; dos Santos, Hannah Lia Ettiene Peruch Lemos; dos Santos, Maíra Moreira; Pinheiro, Filipe Aguera; Navarro, Italmar Teodorico; Garcia, João Luis; Freire, Roberta Lemos

2017-01-01

The aims of this study were to verify the prevalence of Cryptosporidium spp. and Giardia spp. in animal feces and drinking water on dairy farms and to identify a possible relation between the exposure factors and the presence of these parasites. Fecal samples from cattle and humans and water samples were collected on dairy farms in Paraná, Brazil. Analysis of (oo)cysts in the feces was performed by the modified Ziehl-Neelsen staining and centrifugal flotation in zinc sulfate. Test-positive samples were subjected to nested PCR amplification of the 18SSU ribosomal RNA gene for identification of Cryptosporidium and Giardia and of the gp60 gene for subtyping of Cryptosporidium. Microbiological analysis of water was carried out by the multiple-tube method and by means of a chromogenic substrate, and parasitological analysis was performed on 31 samples by direct immunofluorescence and nested PCR of the genes mentioned above. Identification of the species of Cryptosporidium was performed by sequencing and PCR with analysis of restriction fragment length polymorphisms. The prevalence of Giardia and Cryptosporidium was higher in calves than in adults. Among the samples of cattle feces, Cryptosporidium parvum was identified in 41 (64%), C. ryanae in eight (12.5%), C. bovis in four (6.3%), C. andersoni in five (7.8%), and a mixed infection in 20 samples (31.3%). These parasites were not identified in the samples of human feces. Thermotolerant coliform bacteria were identified in 25 samples of water (45.5%). Giardia duodenalis and C. parvum were identified in three water samples. The gp60 gene analysis of C. parvum isolates revealed the presence of two strains (IIaA20G1R1 and IIaA17G2R2) in the fecal samples and one (IIaA17G2R1) in the water samples. The presence of coliforms was associated with the water source, structure and degradation of springs, rain, and turbidity. The prevalence of protozoa was higher in calves up to six months of age. C. parvum and G. duodenalis were identified in the water of dairy farms, as were thermotolerant coliforms; these findings point to the need for guidance on handling of animals, preservation of water sources, and water treatment. PMID:28403147

Cryptosporidium spp. and Giardia spp. in feces and water and the associated exposure factors on dairy farms.

PubMed

Toledo, Roberta Dos Santos; Martins, Felippe Danyel Cardoso; Ferreira, Fernanda Pinto; de Almeida, Jonatas Campos; Ogawa, Liza; Dos Santos, Hannah Lia Ettiene Peruch Lemos; Dos Santos, Maíra Moreira; Pinheiro, Filipe Aguera; Navarro, Italmar Teodorico; Garcia, João Luis; Freire, Roberta Lemos

2017-01-01

The aims of this study were to verify the prevalence of Cryptosporidium spp. and Giardia spp. in animal feces and drinking water on dairy farms and to identify a possible relation between the exposure factors and the presence of these parasites. Fecal samples from cattle and humans and water samples were collected on dairy farms in Paraná, Brazil. Analysis of (oo)cysts in the feces was performed by the modified Ziehl-Neelsen staining and centrifugal flotation in zinc sulfate. Test-positive samples were subjected to nested PCR amplification of the 18SSU ribosomal RNA gene for identification of Cryptosporidium and Giardia and of the gp60 gene for subtyping of Cryptosporidium. Microbiological analysis of water was carried out by the multiple-tube method and by means of a chromogenic substrate, and parasitological analysis was performed on 31 samples by direct immunofluorescence and nested PCR of the genes mentioned above. Identification of the species of Cryptosporidium was performed by sequencing and PCR with analysis of restriction fragment length polymorphisms. The prevalence of Giardia and Cryptosporidium was higher in calves than in adults. Among the samples of cattle feces, Cryptosporidium parvum was identified in 41 (64%), C. ryanae in eight (12.5%), C. bovis in four (6.3%), C. andersoni in five (7.8%), and a mixed infection in 20 samples (31.3%). These parasites were not identified in the samples of human feces. Thermotolerant coliform bacteria were identified in 25 samples of water (45.5%). Giardia duodenalis and C. parvum were identified in three water samples. The gp60 gene analysis of C. parvum isolates revealed the presence of two strains (IIaA20G1R1 and IIaA17G2R2) in the fecal samples and one (IIaA17G2R1) in the water samples. The presence of coliforms was associated with the water source, structure and degradation of springs, rain, and turbidity. The prevalence of protozoa was higher in calves up to six months of age. C. parvum and G. duodenalis were identified in the water of dairy farms, as were thermotolerant coliforms; these findings point to the need for guidance on handling of animals, preservation of water sources, and water treatment.

Impacts of global change on the concentrations and dilution of combined sewer overflows in a drinking water source.

PubMed

Jalliffier-Verne, Isabelle; Leconte, Robert; Huaringa-Alvarez, Uriel; Madoux-Humery, Anne-Sophie; Galarneau, Martine; Servais, Pierre; Prévost, Michèle; Dorner, Sarah

2015-03-01

This study presents an analysis of climate change impacts on a large river located in Québec (Canada) used as a drinking water source. Combined sewer overflow (CSO) effluents are the primary source of fecal contamination of the river. An analysis of river flowrates was conducted using historical data and predicted flows from a future climate scenario. A spatio-temporal analysis of water quality trends with regard to fecal contamination was performed and the effects of changing flowrates on the dilution of fecal contaminants were analyzed. Along the river, there was a significant spatial trend for increasing fecal pollution downstream of CSO outfalls. Escherichia coli concentrations (upper 95th percentile) increased linearly from 2002 to 2012 at one drinking water treatment plant intake. Two critical periods in the current climate were identified for the drinking water intakes considering both potential contaminant loads and flowrates: local spring snowmelt that precedes river peak flow and extra-tropical storm events that occur during low flows. Regionally, climate change is expected to increase the intensity of the impacts of hydrological conditions on water quality in the studied basin. Based on climate projections, it is expected that spring snowmelt will occur earlier and extreme spring flowrates will increase and low flows will generally decrease. High and low flows are major factors related to the potential degradation of water quality of the river. However, the observed degradation of water quality over the past 10 years suggests that urban development and population growth may have played a greater role than climate. However, climate change impacts will likely be observed over a longer period. Source water protection plans should consider climate change impacts on the dilution of contaminants in addition to local land uses changes in order to maintain or improve water quality. Copyright © 2014 Elsevier B.V. All rights reserved.

Towards establishing a human fecal contamination index in microbial source tracking

EPA Science Inventory

There have been significant advances in development of PCR-based methods to detect source associated DNA sequences (markers), but method evaluation has focused on performance with individual challenge samples. Little attention has been given to integration of multiple samples fro...

Characterization of Vancomycin-Resistant Enterococcus faecalis and Enterococcus faecium Isolated from Fresh Produces and Human Fecal Samples.

PubMed

Kim, Min-Chan; Cha, Min-Hyeok; Ryu, Jae-Gee; Woo, Gun-Jo

2017-04-01

Increased enterococcal infections in hospitals and multidrug-resistant and vancomycin-resistant enterococci (VRE) isolated from humans, animals, and food sources raised public health concern on the presence of VRE in multiple sources. We performed a comparative analysis of the antimicrobial resistance and genetics of VRE isolates derived from fresh produce and human fecal samples. Of 389 Enterococcus isolates, 8 fecal and 3 produce isolates were resistant to vancomycin and teicoplanin; all harbored vanA gene. The VRE isolates showed multidrug-resistant properties. The isolates from fresh produce in this study showed to have the common shared characteristics with the isolates from humans by the results of antimicrobial resistance, multilocus sequence typing, and Tn 1546 transposon analysis. Therefore, VRE isolates from fresh produce are likely related to VRE derived from humans. The results suggested that VRE may contaminate vegetables through the environment, and the contaminated vegetables could then act as a vehicle for human infections. Ongoing nationwide surveillance of antibiotic resistance and the promotion of the proper use of antibiotics are necessary.

Reconnaissance of water-quality characteristics of streams in the City of Charlotte and Mecklenburg County, North Carolina

USGS Publications Warehouse

Eddins, W.H.; Crawford, J.K.

1984-01-01

In 1979-81, water samples were collected from 119 sites on streams throughout the City of Charlotte and Mecklenburg County, North Carolina, and were analyzed for specific conductance, dissolved chloride, hardness, pH, total alkalinity, total phosphorus, trace elements, arsenic, cadmium, chromium, copper, iron, lead, manganese, mercury, silver, and zinc and biological measures including dissolved oxygen, biochemical oxygen demand, fecal coliform bacteria, and fecal streptococcus bacteria. Sampling was conducted during both low flow (base flow) and high flow. Several water-quality measures including pH, total arsenic, total cadmium, total chromium, total copper, total iron, total lead, total manganese, total mercury, total silver, total zinc, dissolved oxygen, and fecal coliform bacteria at times exceeded North Carolina water-quality standards in various streams. Runoff from non-point sources appears to contribute more to the deterioration of streams in Charlotte and Mecklenburg County than point-source effluents. Urban and industrial areas contribute various trace elements. Residential and rural areas and municipal waste-water treatment plants contribute high amounts of phosphorus.

Intestinal Fluid Permeability in Atlantic Salmon (Salmo salar L.) Is Affected by Dietary Protein Source.

PubMed

Hu, Haibin; Kortner, Trond M; Gajardo, Karina; Chikwati, Elvis; Tinsley, John; Krogdahl, Åshild

2016-01-01

In Atlantic salmon (Salmo salar L.), and also in other fish species, certain plant protein ingredients can increase fecal water content creating a diarrhea-like condition which may impair gut function and reduce fish growth. The present study aimed to strengthen understanding of the underlying mechanisms by observing effects of various alternative plant protein sources when replacing fish meal on expression of genes encoding proteins playing key roles in regulation of water transport across the mucosa of the distal intestine (DI). A 48-day feeding trial was conducted with five diets: A reference diet (FM) in which fish meal (72%) was the only protein source; Diet SBMWG with a mix of soybean meal (30%) and wheat gluten (22%); Diet SPCPM with a mix of soy protein concentrate (30%) and poultry meal (6%); Diet GMWG with guar meal (30%) and wheat gluten (14.5%); Diet PM with 58% poultry meal. Compared to fish fed the FM reference diet, fish fed the soybean meal containing diet (SBMWG) showed signs of enteritis in the DI, increased fecal water content of DI chyme and higher plasma osmolality. Altered DI expression of a battery of genes encoding aquaporins, ion transporters, tight junction and adherens junction proteins suggested reduced transcellular transport of water as well as a tightening of the junction barrier in fish fed the SBMWG diet, which may explain the observed higher fecal water content and plasma osmolality. DI structure was not altered for fish fed the other experimental diets but alterations in target gene expression and fecal water content were observed, indicating that alterations in water transport components may take place without clear effects on intestinal structure.

Sanitary quality of the Jordan River in Salt Lake County, Utah

USGS Publications Warehouse

Thompson, K.R.

1984-01-01

This investigation of the sanitary quality of the Jordan River was conducted from July 1980 to October 1982 using indicator bacteria rather than specific pathogens. A serious sanitary problem was identified. Concentrations of total coliform bacteria often exceeded 5,000 colonies per 100 milliliters and concentrations of fecal coliform bacteria often exceeded 2,000 colonies per 100 milliliters in the lower reaches of the river. At times these levels were greatly exceeded. The most conspicuous aspect of the bacteriological data is its extreme variability. Seven waste-water treatment plants, seven major tributaries, numerous storm conduits, irrigation-return flow, and other sources all contribute to the dynamic system that determines the sanitary quality of the river. Because of this variability the sanitary quality of the river cannot be predicted at any one time. In general, concentrations of all three indicator bacteria increased in a downstream direction. Storm runoff from urban areas contributed large concentrations of indicator bacteria to the river. Regression analysis of 9 years of data collected at 1700 South Street showed a significant positive correlation between both fecal coliform and fecal streptococcal concentrations versus time. Concentrations of fecal coliform and fecal streptococci have both been increasing since 1974 at 1700 South Street. (USGS)

Metabolite identification in fecal microbiota transplantation mouse livers and combined proteomics with chronic unpredictive mild stress mouse livers.

PubMed

Li, Bo; Guo, Kenan; Zeng, Li; Zeng, Benhua; Huo, Ran; Luo, Yuanyuan; Wang, Haiyang; Dong, Meixue; Zheng, Peng; Zhou, Chanjuan; Chen, Jianjun; Liu, Yiyun; Liu, Zhao; Fang, Liang; Wei, Hong; Xie, Peng

2018-01-31

Major depressive disorder (MDD) is a common mood disorder. Gut microbiota may be involved in the pathogenesis of depression via the microbe-gut-brain axis. Liver is vulnerable to exposure of bacterial products translocated from the gut via the portal vein and may be involved in the axis. In this study, germ-free mice underwent fecal microbiota transplantation from MDD patients and healthy controls. Behavioral tests verified the depression model. Metabolomics using gas chromatography-mass spectrometry, nuclear magnetic resonance, and liquid chromatography-mass spectrometry determined the influence of microbes on liver metabolism. With multivariate statistical analysis, 191 metabolites were distinguishable in MDD mice from control (CON) mice. Compared with CON mice, MDD mice showed lower levels for 106 metabolites and higher levels for 85 metabolites. These metabolites are associated with lipid and energy metabolism and oxidative stress. Combined analyses of significantly changed proteins in livers from another depression model induced by chronic unpredictive mild stress returned a high score for the Lipid Metabolism, Free Radical Scavenging, and Molecule Transports network, and canonical pathways were involved in energy metabolism and tryptophan degradation. The two mouse models of depression suggest that changes in liver metabolism might be involved in the pathogenesis of MDD. Conjoint analyses of fecal, serum, liver, and hippocampal metabolites from fecal microbiota transplantation mice suggested that aminoacyl-tRNA biosynthesis significantly changed and fecal metabolites showed a close relationship with the liver. These findings may help determine the biological mechanisms of depression and provide evidence about "depression microbes" impacting on liver metabolism.

Characterization of the estrous cycle and assessment of reproductive status in Matschie's tree kangaroo (Dendrolagus matschiei) with fecal progestin profiles.

PubMed

North, Lindsay A; Harder, John D

2008-03-01

The population of Matschie's tree kangaroos (Dendrolagus matschiei) held in North American zoos has declined to critically low numbers, and information on the reproductive biology of tree kangaroos is limited. The objectives of this study were to (1) characterize the temporal features of the estrous cycle through the measurement of fecal progesterone metabolite (i.e., progestin) concentrations and (2) determine the reproductive status of female tree kangaroos in the captive population of North America through the identification of estrous cyclicity. Fecal pellets and observations of estrous behaviors were collected from 16 captive female tree kangaroos. Fecal pellets were sampled and extracted with methanol, and progestin concentrations were quantified using a radioimmunoassay (RIA) for progesterone and its metabolites. A progestin profile was obtained for each female by plotting fecal progestin concentrations for every third day over a 120-day period. Profiles for 12 of 16 females showed evidence of estrous cyclicity (P<0.01). The mean length of the estrous cycle was estimated at 58.9+/-2.4 days (n=11). Progestin concentrations were low during the first 15-20 days of the luteal phase and remained elevated above baseline only during the last 30.2+/-3.2 days of the luteal phase, which averaged 46.6+/-2.5 days in duration. The progestin profile observed in the estrous cycle of Matschie's tree kangaroos in this study is very similar to that seen in the non-pregnant cycle of several other species in the family Macropodidae.

Norovirus GII.Pe Genotype: Tracking a Foodborne Outbreak on a Cruise Ship Through Molecular Epidemiology, Brazil, 2014.

PubMed

Morillo, Simone Guadagnucci; Luchs, Adriana; Cilli, Audrey; Ribeiro, Cibele Daniel; de Cássia Compagnoli Carmona, Rita; do Carmo Sampaio Tavares Timenetsky, Maria

2017-06-01

Norovirus (NoV) is recognized as the most common cause of foodborne outbreaks. In 2014, an outbreak of acute gastroenteritis occurred on a cruise ship in Brazil, and NoV became the suspected etiology. Here we present the molecular identification of the NoV strains and the use of sequence analysis to determine modes of virus transmission. Food (cream cheese, tuna salad, grilled fish, orange mousse, and vegetables soup) and clinical samples were analyzed by ELISA, conventional RT-PCR, qRT-PCR, and sequencing. Genogroup GII NoV was identified by ELISA and conventional RT-PCR in fecal samples from 5 of 12 patients tested (41.7%), and in the orange mousse food sample by conventional RT-PCR and qRT-PCR. Two fecal GII NoV samples and the orange mousse GII NoV sample were successfully genotyped as GII.Pe (ORF 1), revealed 98.0-98.8% identities among them, and shared phylogenetically distinct cluster. Establishing the source of a NoV outbreak can be a challenging task. In this report, the molecular analysis of the partial RdRp NoV gene provided a powerful tool for genotyping (GII.Pe) and tracking of outbreak-related samples. In addition, the same fast and simple extraction methods applied to clinical samples could be successfully used for complex food matrices, and have the potential to be introduced in routine laboratories for screening foods for presence of NoV.

In vivo anthelmintic activity of Anogeissus leiocarpus Guill & Perr (Combretaceae) against nematodes in naturally infected sheep.

PubMed

Soro, Dramane; Koné, Witabouna Mamidou; Bonfoh, Bassirou; Dro, Bernadin; Toily, Kassédo Bénédicte; Kamanzi, Kagoyire

2013-07-01

The identification of new anthelmintic drugs becomes a priority because of the availability of a handful of drugs, cost of treatments, and recent emergence of drug resistance. Medicinal plants are a good source of bioactive compounds for development of drugs. In this study, in vivo efficacy of Anogeissus leiocarpus was assessed in sheep naturally infected with gastrointestinal nematodes. Fecal examination, serological analyses, and necropsy were carried out to determine the egg and worm-burden reduction. The administration of ethanolic extract (single oral dose of 80 mg/kg) of A. leiocarpus induced a moderate fecal egg reduction (81 %) and adult worm-burden reduction (87 %) against Haemonchus contortus and Trichostrongylus colubriformis (82 %). The plant exhibited high efficacy against adult Strongyloïdes papillosus (100 %), Gaigeria pachyscelis (90 %), Cooperia curticei (100 %), and Oesophagostomum columbianum (95 %) but low efficacy against Trichostrongylus axei (67 %) and Trichuris globulosa (79 %). All these helminthes were sensitive to fenbendazole, except O. columbianum which showed a decrease susceptibility (17 %). The plant extract also improved certain biological parameters by increasing bodyweight from 0.7 ± 2.9 to 3.3 ± 1.9 % and improving hematocrit of 6.9 ± 1.6 % 3-week posttreatment. It emerges from the results that the plant possesses significant effectiveness on diarrhea; all treated animals gave normal feces. This study has shown that A. leiocarpus could find an application in the control of multiparasitism in small ruminants.

Reconnaissance of the hydrology, water quality, and sources of bacterial and nutrient contamination in the Ozark Plateaus aquifer system and Cave Springs Branch of Honey Creek, Delaware County, Oklahoma, March 1999-March 2000

USGS Publications Warehouse

Schlottmann, Jamie L.; Tanner, Ralph S.; Samadpour, Mansour

2000-01-01

A reconnaissance investigation of hydrology and water quality was conducted to evaluate possible sources of bacteria and nutrient contamination in the Cave Springs Branch basin and the underlying karstic Ozark Plateau aquifer system. Objectives were to: (1) determine the directions of ground-water flow in the basin and determine whether Cave Springs Branch interacts with ground water, (2) compare water quality in Cave Springs Branch with water quality in nearby wells to determine whether the stream is contaminating nearby wells, and (3) determine sources of fecal coliform bacteria and nitrate contamination in Cave Springs Branch and ground water. Potential sources of bacteria and nitrate in the area include cultivated agriculture, cow and horse on pasture, poultry production, households, and wildlife. Presence of fecal coliform and fecal streptococcal bacteria directly indicate fecal contamination and the potential for the presence of other pathogenic organisms in a water supply. Nitrate in drinking water poses health risks and may indicate the presence of additional contaminants. Fecal coliform bacteria colony counts were least in wells, intermediate in the poultry-processing plant wastewater outfall and Honey Creek above the confluence with Cave Springs Branch, and greatest in Cave Springs Branch. Bacteria strains and resistance to antibiotics by some bacteria indicate that livestock may have been sources of some bacteria in the water samples. Multiple antibiotic resistances were not present in the isolates from the water samples, indicating that the bacteria may not be from human or poultry sources. Ribotyping indicates that Escherichia coli bacteria in water samples from the basin were from bird, cow, horse, dog, deer, and human sources. The presence of multiple ribotypes from each type of animal source except bird indicates that most of the bacteria are from multiple populations of source animals. Identifiable sources of bacteria in Cave Springs Branch at the state line were dominantly cow and horse with one ribotype from bird. Escherichia coli was detected in only one well sample. Bacterial ribotypes in water from that upgradient well indicated human and dog feces as sources for bacteria, and that on site wastewater treatment may not always be adequate in these highly permeable soils. Greater concentrations of nitrate in Cave Springs Branch and O'Brien Spring relative to the poultry-processing plant wastewater outfall may be due, in part, to conversion of ammonia from poultry processing plant wastewater. The poultry-processing plant wastewater outfall sample collected in March 2000 contained greater concentrations of ammonia and total organic nitrogen plus ammonia than the spring, stream, and well samples collected during August 1999. Cave Springs Branch and Honey Creek contributed approximately equal loads of nitrogen to Honey Creek below the confluence and the greatest loads of nitrogen were introduced to Cave Springs Branch by the poultry processing plant wastewater outfall and O'Brien Spring. Nitrate concentrations in upgradient well samples ranged from 0.38 to 4.60 milligrams per liter, indicating that there are sources of ground-water nitrogen other than Cave Springs Branch, such as animal waste, fertilizer, or human waste. Nitrogen compounds in water from wells downgradient of Cave Springs Branch may be from Cave Springs Branch, fertilizers, animal waste, or human waste.

The presence and near-shore transport of human fecal pollution in Lake Michigan beaches

USGS Publications Warehouse

Molloy, S.L.; Liu, L.B.; Phanikumar, M.S.; Jenkins, T.M.; Wong, M.V.; Rose, J.B.; Whitman, R.L.; Shively, D.A.; Nevers, M.B.

2005-01-01

The Great Lakes are a source of water for municipal, agricultural and industrial use, and support significant recreation, commercial and sport fishing industries. Every year millions of people visit the 500 plus recreational beaches in the Great Lakes. An increasing public health risk has been suggested with increased evidence of fecal contamination at the shoreline. To investigate the transport and fate of fecal pollution at Great Lakes beaches and the health risk associated with swimming at these beaches, the near-shore waters of Mt Baldy Beach, Lake Michigan and Trail Creek, a tributary discharging into the lake were examined for fecal pollution indicators. A model of surf zone hydrodynamics coupled with a transport model with first-order inactivation of pollutant was used to understand the relative importance of different processes operating in the surf zone (e.g. physical versus biological processes). The Enterococcus human fecal pollution marker, which targets a putative virulence factor, the enterococcal surface protein (esp) in Enterococcus faecium, was detected in 2/28 samples (7%) from the tributaries draining into Lake Michigan and in 6/30 samples (20%) from Lake Michigan beaches. Preliminary analysis suggests that the majority of fecal indicator bactateria variation and water quality changes at the beaches can be explained by inputs from the influential stream and hydrometeorological conditions. Using modeling methods to predict impaired water quality may help reduce potential health threats to recreational visitors.

Removal of bacterial fecal indicators, coliphages and enteric adenoviruses from waters with high fecal pollution by slow sand filtration.

PubMed

Bauer, Rosalie; Dizer, Halim; Graeber, Ingeborg; Rosenwinkel, Karl-Heinz; López-Pila, Juan M

2011-01-01

The aim of the present study was to estimate the performance of slow sand filtration (SSF) facilities, including the time needed for reaching stabilization (maturation), operated with surface water bearing high fecal contamination, representing realistic conditions of rivers in many emerging countries. Surface water spiked with wastewater was infiltrated at different pore water velocities (PWV) and samples were collected at different migration distances. The samples were analyzed for phages and to a lesser extent for fecal bacteria and enteric adenoviruses. At the PWV of 50 cm/d, at which somatic phages showed highest removal, their mean log(10) removal after 90 cm migration was 3.2. No substantial differences of removal rates were observed at PWVs between 100 and 900 cm/d (2.3 log(10) mean removal). The log(10) mean removal of somatic phages was less than the observed for fecal bacteria and tended more towards that of enteric adenoviruses This makes somatic phages a potentially better process indicator than Escherichia coli for the removal of viruses in SSF. We conclude that SSF, and by inference in larger scale river bank filtration (RBF), is an excellent option as a component in multi-barrier systems for drinking water treatment also in areas where the sources of raw water are considerably fecally polluted, as often found in many emerging countries. Copyright © 2010 Elsevier Ltd. All rights reserved.

Potential Regrowth and Recolonization of Salmonellae and Indicators in Biosolids and Biosolid-Amended Soil

PubMed Central

Zaleski, Kathleen J.; Josephson, Karen L.; Gerba, Charles P.; Pepper, Ian L.

2005-01-01

This study evaluated the potential for conversion of Class B to Class A biosolids with respect to salmonellae and fecal coliforms during solar drying in concrete lined drying beds. Anaerobically (8% solids) and aerobically (2% solids) digested Class B biosolids were pumped into field-scale drying beds, and microbial populations and environmental conditions were monitored. Numbers of fecal coliforms and salmonellae decreased as temperature and rate of desiccation increased. After 3 to 4 weeks, Class A requirements were achieved in both biosolids for the pathogens and the indicators. However, following rainfall events, significant increase in numbers was observed for both fecal coliforms and salmonellae. In laboratory studies, regrowth of fecal coliforms was observed in both biosolids and biosolid-amended soil, but the regrowth of salmonellae observed in the concrete-lined drying beds did not occur. These laboratory studies demonstrated that pathogens decreased in numbers when soil was amended with biosolids. Based on serotyping, the increased numbers of salmonellae seen in the concrete lined drying beds following rainfall events was most likely due to recolonization due to contamination from fecal matter introduced by animals and not from regrowth of salmonellae indigenous to biosolids. Overall, we conclude that the use of concrete-lined beds created a situation in which moisture added as rainfall accumulated in the beds, promoting the growth of fecal coliforms and salmonellae added from external sources. PMID:16000779

Effects of Holding Time, Storage, and the Preservation of ...

EPA Pesticide Factsheets

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 that were used to collect fecal indicators could be stored frozen and analyzed at a later date and the second part was to determine if refrigerated water samples could be held for 24 to 48 hours prior to analysis by qPCR. Both of these studies answer questions that were important in the analysis of fresh and marine surface water samples for beach monitoring purposes. 1) Develop and evaluate qPCR assays and test methods for the detection and quantification of genetic markers from indicator bacteria that are associated with human fecal waste and from two new groups of general fecal indicator bacteria (E. coli and Clostridia) that historically have been widely used or are favored in specific regions 2) Determine the occurrence and densities of genetic markers detected by new qPCR assays developed under objective 1 and compare with occurrence and densities of genetic markers detected by previously developed qPCR assays for enterococci and total Bacterioidalesin waste waters and fecal material from different animal sources. 3) Determine stability of fecal indicator bacteria target DNA sequences in freezer archived filter retentates of ambient surface water samples 4) Determine the densitie

Geographic distribution of the muscle-dwelling nematode Parelaphostrongylus odocoilei in North America, using molecular identification of first-stage larvae.

PubMed

Jenkins, Emily J; Appleyard, Greg D; Hoberg, Eric P; Rosenthal, Benjamin M; Kutz, Susan J; Veitch, Alasdair M; Schwantje, Helen M; Elkin, Brett T; Polley, Lydden

2005-06-01

Molecular identification of dorsal-spined larvae (DSL) from fecal samples indicates that the protostrongylid parasite Parelaphostrongylus odocoilei occupies a broader geographic range in western North America than has been previously reported. We analyzed 2,124 fecal samples at 29 locations from thinhorn sheep (Ovis dalli dalli and O. d. stonei), bighorn sheep (Ovis canadensis canadensis and O. c. californiana), mountain goats (Oreamnos americanus), woodland caribou (Rangifer tarandus caribou), mule deer (Odocoileus hemionus hemionus), and black-tailed deer (O. h. columbianus). The DSL were recovered from populations of thinhorn sheep south, but not north, of the Arctic Circle, and they were not recovered from any of the bighorn sheep populations that we examined. In total, DSL were recovered from 20 locations in the United States and Canada (Alaska, Yukon Territory, Northwest Territories, British Columbia, Alberta, and California). The DSL were identified as P. odocoilei by comparing sequences of the second internal transcribed spacer (ITS2) region of ribosomal RNA among 9 protostrongylid species validated by adult comparative morphology. The ITS2 sequences were markedly different between Parelaphostrongylus and other protostrongylid genera. Smaller fixed differences served as diagnostic markers for the 3 species of Parelaphostrongylus. The ITS2 sequences (n = 60) of P. odocoilei were strongly conserved across its broad geographic range from California to Alaska. Polymorphism at 5 nucleotide positions was consistent with multiple copies of the ITS2 within individual specimens of P. odocoilei. This work combines extensive fecal surveys, comparative morphology, and molecular diagnostic techniques to describe comprehensively the host associations and geographic distribution of a parasitic helminth.

Use of Direct LAMP Screening of Broiler Fecal Samples for Campylobacter jejuni and Campylobacter coli in the Positive Flock Identification Strategy.

PubMed

Sabike, Islam I; Uemura, Ryoko; Kirino, Yumi; Mekata, Hirohisa; Sekiguchi, Satoshi; Okabayashi, Tamaki; Goto, Yoshitaka; Yamazaki, Wataru

2016-01-01

Rapid identification of Campylobacter -positive flocks before slaughter, following freezing and heat treatment for the Campylobacter -positive carcasses at the slaughterhouses is an effective control strategy against foodborne campylobacteriosis. We evaluated a loop-mediated isothermal amplification (LAMP) assay for the direct screening of naturally contaminated chicken cloacal swabs for C. jejuni / C. coli to compare this assay with conventional quantitative culture methods. In a comparison study of 165 broilers, the LAMP assay showed 82.8% (48/58 by conventional culture) sensitivity, 100% (107/107) specificity, 100% (48/48) positive predictive value (PPV), and 91.5% (107/117) negative predictive value (NPV). In a comparison of 55 flocks, LAMP showed 90.5% (19/21) sensitivity, 100% (34/34) specificity, 100% (19/19) PPV, and 94.4% (34/36) NPV. In the cumulative total of 28 farm-level comparisons, LAMP showed 100% (12/12) sensitivity, 100% (16/16) specificity, 100% (12/12) PPV, and 100% (16/16) NPV. The LAMP assay required less than 90 min from the arrival of the fecal samples to final results in the laboratory. This suggests that the LAMP assay will facilitate the identification of C. jejuni / C. coli -positive broiler flocks at the farm level or in slaughterhouses before slaughtering, which would make it an effective tool in preventing the spread of Campylobacter contamination.

Development of quantitative PCR assays targeting 16S rRNA gene of Enterococcus spp. and their application to the identification of Enterococcus species in environmental samples

EPA Science Inventory

The detection of environmental enterococci has primarily been determined using culture-based techniques that might exclude some enterococci species as well as those that are nonculturable. To address this, the relative abundance of enterococci was examined by challenging fecal an...

Identification of Termite Species by the Hydrocarbons in their Feces

Treesearch

Michael I. Haverty; R. Joseph Woodrow; Lori J. Nelson; J. Kenneth Grace

2005-01-01

Blends of abundant cuticular hydrocarbons are species-specific for termites (Isoptera) and can be used to identify a given taxon without the diagnostic castes, soldiers or adults. We demonstrate that hydrocarbon extracts of termite fecal pellets from damaged wood can also be characterized and used to identify termites responsible for damage, even though termites are no...

Distributions of Fecal Markers in Wastewater from Different Climatic Zones for Human Fecal Pollution Tracking in Australian Surface Waters

PubMed Central

Sidhu, J. P. S.; Smith, K.; Beale, D. J.; Gyawali, P.; Toze, S.

2015-01-01

Recreational and potable water supplies polluted with human wastewater can pose a direct health risk to humans. Therefore, sensitive detection of human fecal pollution in environmental waters is very important to water quality authorities around the globe. Microbial source tracking (MST) utilizes human fecal markers (HFMs) to detect human wastewater pollution in environmental waters. The concentrations of these markers in raw wastewater are considered important because it is likely that a marker whose concentration is high in wastewater will be more frequently detected in polluted waters. In this study, quantitative PCR (qPCR) assays were used to determine the concentrations of fecal indicator bacteria (FIB) Escherichia coli and Enterococcus spp., HFMs Bacteroides HF183, human adenoviruses (HAdVs), and polyomaviruses (HPyVs) in raw municipal wastewater influent from various climatic zones in Australia. E. coli mean concentrations in pooled human wastewater data sets (from various climatic zones) were the highest (3.2 × 106 gene copies per ml), followed by those of HF183 (8.0 × 105 gene copies per ml) and Enterococcus spp. (3.6 × 105 gene copies per ml). HAdV and HPyV concentrations were 2 to 3 orders of magnitude lower than those of FIB and HF183. Strong positive and negative correlations were observed between the FIB and HFM concentrations within and across wastewater treatment plants (WWTPs). To identify the most sensitive marker of human fecal pollution, environmental water samples were seeded with raw human wastewater. The results from the seeding experiments indicated that Bacteroides HF183 was more sensitive for detecting human fecal pollution than HAdVs and HPyVs. Since the HF183 marker can occasionally be present in nontarget animal fecal samples, it is recommended that HF183 along with a viral marker (HAdVs or HPyVs) be used for tracking human fecal pollution in Australian environmental waters. PMID:26682850

Understanding beach health throughout the Great Lakes-Entering a new era of investigations

USGS Publications Warehouse

,

2010-01-01

For over a decade, the U.S. Geological Survey (USGS) has been a leader in the science of beach health. The overall mission of this work is to provide science-based information and methods that will allow beach managers to more accurately make beach closure and advisory decisions, understand the sources and physical processes affecting beach contaminants, and understand how science-based information can be used to mitigate and restore beaches and protect the public. The work consists of four science elements-real-time assessments; pathogens and microbial source tracking; coastal processes; and data analysis, interpretation, and communication - which are described in this fact sheet. Some of the key questions for USGS beach research are the following: Are there better ways to inform the public whether they can use a beach without risking their health? How do new rapid analytical methods compare to traditional methods for determining concentrations of fecal-indicator bacteria at beaches? Are pathogens present at beaches and, if so, how do they get to the beach, and what is their source? How do sand movement and wave action on the beach affect fecal-indicator-bacteria and pathogen concentrations in the lake water? What are the best indicators of pathogenic microorganisms? With so many potential sources of fecal contamination at a beach, what methods can be used to distinguish the contributions from humans? What characteristics of beaches contribute most to influencing bacterial indicator and pathogen concentrations in beach sands and groundwater?

Evaluation of sewage source and fate on southeast Florida coastal reefs

USGS Publications Warehouse

Carrie, Futch J.; Griffin, Dale W.; Banks, K.; Lipp, E.K.

2011-01-01

Water, sponge and coral samples were collected from stations impacted by a variety of pollution sources and screened for human enteric viruses as conservative markers for human sewage. While human enteroviruses and adenoviruses were not detected, noroviruses (NoV; human genogroups I and II) were detected in 31% of samples (especially in sponge tissue). Stations near inlets were the only ones to show multiple sample types positive for NoV. Fecal indicator bacteria and enteric viruses were further evaluated at multiple inlet stations on an outgoing tide. Greatest indicator concentrations and highest prevalence of viruses were found at the mouth of the inlet and offshore in the inlet plume. Results suggest that inlets moving large volumes of water into the coastal zone with tides may be an important source of fecal contaminants. Efforts to reduce run-off or unintended release of water into the Intracoastal Waterway may lower contaminants entering sensitive coastal areas. ?? 2011 Elsevier Ltd.

40 CFR 141.402 - Ground water source microbial monitoring and analytical methods.

Code of Federal Regulations, 2011 CFR

2011-07-01

... approves the use of E. coli as a fecal indicator for source water monitoring under this paragraph (a). If the repeat sample collected from the ground water source is E.coli positive, the system must comply... listed in the in paragraph (c)(2) of this section for the presence of E. coli, enterococci, or coliphage...

DOE Office of Scientific and Technical Information (OSTI.GOV)

Andersen, Gary L.; Dubinsky, Eric A.

Herein are described 1058 different bacterial taxa that were unique to either human, grazing mammal, or bird fecal wastes. These identified taxa can serve as specific identifier taxa for these sources in environmental waters. Two field tests in marine waters demonstrate the capacity of phylogenetic microarray analysis to track multiple sources with one test.

Effect of phytoplankton on Escherichia coli survival in laboratory microcosms

USDA-ARS?s Scientific Manuscript database

Fecal contamination of water sources is an important water quality issue for agricultural irrigation ponds. Escherichia coli is a common microbial indicator used to evaluate recreational and irrigation water quality. Nuisance algae commonly grow in low- or no-flow irrigation water source The objecti...

Relationships and trends of E. Coli, human-associated Bacteroides, and pathogens in the Proctor Creek Watershed

EPA Science Inventory

Urban surface waters can be impacted by anthropogenic sources such as impervious surfaces, sanitary and storm sewers, and failing infrastructure. Fecal indicator bacteria (FIB) and microbial source tracking (MST) markers are common gauges of stream water quality, however, little...

Effects of breed, sex, and age on the variation and ability of fecal near-infrared reflectance spectra to predict the composition of goat diets.

PubMed

Walker, J W; Campbell, E S; Lupton, C J; Taylor, C A; Waldron, D F; Landau, S Y

2007-02-01

The effects of breed, sex, and age of goats on fecal near-infrared reflectance spectroscopy-predicted percentage juniper in the diet were investigated, as were spectral differences in feces from goats differing in estimated genetic merit for juniper consumption. Eleven goats from each breed, sex, and age combination, representing 2 breeds (Angora and meat-type), 3 sex classifications (female, intact male, and castrated male), and 2 age categories [adult and kid (less than 12 mo of age)] were fed complete, pelleted rations containing 0 or 14% juniper. After 7 d on the same diet, fecal samples were collected for 3 d, and the spectra from the 3 replicate samples were averaged. Fecal samples were assigned to calibration or validation data sets. In a second experiment, Angora and meat goats with high or low estimated genetic merit for juniper consumption were fed the same diet to determine the effect of consumer group on fecal spectra. Feces were scanned in the 1,100- to 2,500-nm range with a scanning reflectance monochromator. Fecal spectra were analyzed for the difference in spectral characteristics and for differences in predicted juniper in the diet using internal and independent calibration equations. Internal calibration had a high precision (R(2) = 0.94), but the precision of independent validations (r(2) = 0.56) was low. Spectral differences were affected by diet, sex, breed, and age (P < 0.04). However, diet was the largest source of variation in spectral differences. Predicted percentage of juniper in the diet also showed that diet was the largest source of variation, accounting for 95% of the variation in predictions from internal calibrations and 51% of the variation in independent validations. Predictions from independent calibrations readily detected differences (P < 0.001) in the percentage of juniper in the 2 diets, and the predicted differences were similar to the actual differences. Predicted juniper in the diet was also affected by sex. Feces from goats from different juniper consumer groups fed a common diet were spectrally different, and the difference may have resulted from a greater intake by high- compared with low-juniper-consuming goats. Fecal near-infrared reflectance spectroscopy predictions of botanical composition of diets should be considered an interval scale of measurement.

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 because the presence of oxygen significantly affected the viability and persistence of these obligate anaerobes. In conclusion, measuring Bacteroidales DNA in viable cells is recommended in applied MST studies because extracellular Bacteroidales DNA persists longer in the environment. The methods and results presented are helpful to improve the accuracy of MST applications, to develop a model of fate and transport of host-specific Bacteroidales, and to implement management practices to protect water quality.

Identification of an Intestinal Microbiota Signature Associated With Severity of Irritable Bowel Syndrome.

PubMed

Tap, Julien; Derrien, Muriel; Törnblom, Hans; Brazeilles, Rémi; Cools-Portier, Stéphanie; Doré, Joël; Störsrud, Stine; Le Nevé, Boris; Öhman, Lena; Simrén, Magnus

2017-01-01

We have limited knowledge about the association between the composition of the intestinal microbiota and clinical features of irritable bowel syndrome (IBS). We collected information on the fecal and mucosa-associated microbiota of patients with IBS and evaluated whether these were associated with symptoms. We collected fecal and mucosal samples from adult patients who met the Rome III criteria for IBS at a secondary/tertiary care outpatient clinics in Sweden, as well as from healthy subjects. The exploratory set comprised 149 subjects (110 with IBS and 39 healthy subjects); 232 fecal samples and 59 mucosal biopsy samples were collected and analyzed by 16S ribosomal RNA targeted pyrosequencing. The validation set comprised 46 subjects (29 with IBS and 17 healthy subjects); 46 fecal samples, but no mucosal samples, were collected and analyzed. For each subject, we measured exhaled H 2 and CH 4 , oro-anal transit time, and the severity of psychological and gastrointestinal symptoms. Fecal methanogens were measured by quantitative polymerase chain reaction. Numerical ecology analyses and a machine learning procedure were used to analyze the data. Fecal microbiota showed covariation with mucosal adherent microbiota. By using classic approaches, we found no differences in fecal microbiota abundance or composition between patients with IBS vs healthy patients. A machine learning procedure, a computational statistical technique, allowed us to reduce the 16S ribosomal RNA data complexity into a microbial signature for severe IBS, consisting of 90 bacterial operational taxonomic units. We confirmed the robustness of the intestinal microbial signature for severe IBS in the validation set. The signature was able to discriminate between patients with severe symptoms, patients with mild/moderate symptoms, and healthy subjects. By using this intestinal microbiota signature, we found IBS symptom severity to be associated negatively with microbial richness, exhaled CH 4 , presence of methanogens, and enterotypes enriched with Clostridiales or Prevotella species. This microbiota signature could not be explained by differences in diet or use of medications. In analyzing fecal and mucosal microbiota from patients with IBS and healthy individuals, we identified an intestinal microbiota profile that is associated with the severity of IBS symptoms. NCT01252550. Copyright © 2017 AGA Institute. Published by Elsevier Inc. All rights reserved.

Identification of hotspots and trends of fecal surface water pollution in developing countries

NASA Astrophysics Data System (ADS)

Reder, Klara; Flörke, Martina; Alcamo, Joseph

2015-04-01

Water is the essential resource ensuring human life on earth, which can only prosper when water is available and accessible. But of importance is not only the quantity of accessible water but also its quality, which in case of pollution may pose a risk to human health. The pollutants which pose a risk to human health are manifold, covering several groups such as pathogens, nutrients, human pharmaceuticals, heavy metals, and others. With regards to human health, pathogen contamination is of major interest as 4% of all death and 5.7% of disability or ill health in the world can be attributed to poor water supply, sanitation and personal and domestic hygiene. In developing countries, 2.6 billion people lacked access to improved sanitation in 2011. The lack of sanitation poses a risk to surface water pollution which is a threat to human health. A typical indicator for pathogen pollution is fecal coliform bacteria. The objective our study is to assess fecal pollution in the developing regions Africa, Asia and Latin America using the large-scale water quality model WorldQual. Model runs were carried-out to calculate in-stream concentrations and the respective loadings reaching rivers for the time period 1990 to 2010. We identified hotspots of fecal coliform loadings and in-stream concentrations which were further analyzed and ranked in terms of fecal surface water pollution. Main findings are that loadings mainly originate from the domestic sector, thus loadings are high in highly populated areas. In general, domestic loadings can be attributed to the two subsectors domestic sewered and domestic non sewered. The spatial distribution of both sectors varies across catchments. Hotspot pattern of in-stream concentrations are similar to the loadings pattern although they are different in seasonality. As the dilution varies with climate its dilution capacity is high during seasons with high precipitation, which in turn decreases the in-stream concentrations. The fecal pollution is increasing from 1990 to 2010 with increased loadings and larger number of river kilometers with high fecal pollution. Fecal pollution is mainly caused by the domestic sector, and hence, the sanitation type, collection and treatment (level) of collected wastewater are highly important to ensure good quality of water bodies.

Duplex quantitative real-time PCR assay for the detection and discrimination of the eggs of Toxocara canis and Toxocara cati (Nematoda, Ascaridoidea) in soil and fecal samples.

PubMed

Durant, Jean-Francois; Irenge, Leonid M; Fogt-Wyrwas, Renata; Dumont, Catherine; Doucet, Jean-Pierre; Mignon, Bernard; Losson, Bertrand; Gala, Jean-Luc

2012-12-07

Toxocarosis is a zoonotic disease caused by Toxocara canis (T. canis) and/or Toxocara cati (T. cati), two worldwide distributed roundworms which are parasites of canids and felids, respectively. Infections of humans occur through ingestion of embryonated eggs of T. canis or T. cati, when playing with soils contaminated with dogs or cats feces. Accordingly, the assessment of potential contamination of these areas with these roundworms eggs is paramount. A duplex quantitative real-time PCR (2qPCR) targeting the ribosomal RNA gene internal transcribed spacer (ITS2) has been developed and used for rapid and specific identification of T. canis and T. cati eggs in fecal and soil samples. The assay was set up on DNA samples extracted from 53 adult worms including T. canis, T. cati, T. leonina, Ascaris suum (A. suum) and Parascaris equorum (P. equorum). The assay was used to assess the presence of T. cati eggs in several samples, including 12 clean soil samples spiked with eggs of either T. cati or A. suum, 10 actual soil samples randomly collected from playgrounds in Brussels, and fecal samples from cats, dogs, and other animals. 2qPCR results on dogs and cats fecal samples were compared with results from microscopic examination. 2qPCR assay allowed specific detection of T. canis and T. cati, whether adult worms, eggs spiked in soil or fecal samples. The 2qPCR limit of detection (LOD) in spiked soil samples was 2 eggs per g of soil for a turnaround time of 3 hours. A perfect concordance was observed between 2qPCR assay and microscopic examination on dogs and cats feces. The newly developed 2qPCR assay can be useful for high throughput prospective or retrospective detection of T.canis and/or T. cati eggs in fecal samples as well as in soil samples from playgrounds, parks and sandpits.

Global Distribution of Human-Associated Fecal Genetic Markers in Reference Samples from Six Continents

PubMed Central

2018-01-01

Numerous bacterial genetic markers are available for the molecular detection of human sources of fecal pollution in environmental waters. However, widespread application is hindered by a lack of knowledge regarding geographical stability, limiting implementation to a small number of well-characterized regions. This study investigates the geographic distribution of five human-associated genetic markers (HF183/BFDrev, HF183/BacR287, BacHum-UCD, BacH, and Lachno2) in municipal wastewaters (raw and treated) from 29 urban and rural wastewater treatment plants (750–4 400 000 population equivalents) from 13 countries spanning six continents. In addition, genetic markers were tested against 280 human and nonhuman fecal samples from domesticated, agricultural and wild animal sources. Findings revealed that all genetic markers are present in consistently high concentrations in raw (median log10 7.2–8.0 marker equivalents (ME) 100 mL–1) and biologically treated wastewater samples (median log10 4.6–6.0 ME 100 mL–1) regardless of location and population. The false positive rates of the various markers in nonhuman fecal samples ranged from 5% to 47%. Results suggest that several genetic markers have considerable potential for measuring human-associated contamination in polluted environmental waters. This will be helpful in water quality monitoring, pollution modeling and health risk assessment (as demonstrated by QMRAcatch) to guide target-oriented water safety management across the globe. PMID:29570973

Detailed two-dimensional gel proteomic mapping of the feces of the house dust mite Dermatophagoides pteronyssinus and comparison with D. farinae: Reduced trypsin protease content in D. pteronyssinus and different isoforms.

PubMed

Erban, Tomas; Harant, Karel; Hubert, Jan

2017-06-06

Major domestic mite allergens are present in feces. We present a detailed 2D-E-MS/MS proteomic analysis of the Dermatophagoides pteronyssinus feces. Precise cultivation yielded a pure fecal extract. We detected differences in fecal allergens/digestive enzymes between D. pteronyssinus and D. farinae using 2D-E fingerprinting, including unique information on species-specific protease isoforms. Proteomic analysis was performed by 2D-E coupled with MALDI-TOF/TOF identification. The species-specific differences in the fecal extracts of the mites were attributed to trypsin-like proteases known as group 3 allergens. In D. farinae, Der f 3 exhibited high abundance with a pI similar (acidic) to that of the cysteine protease Der f 1 and the chymotrypsin protease Der f 6, whereas in D. pteronyssinus, Der p 3 was rarely detected and exhibited low abundance only at basic pI. Moreover, Der p 9 was detected at a pI of ~ 10, in contrast to Der p 1 and Der p 6, suggesting different compartmentalization in the body. Overall, in D. pteronyssinus feces, allergens of groups 1, 2, 6, and 15 were quantitatively similar to those of D. farinae with the exception of the group 3 and 9 allergens. This work provides novel insights into mite-defecated proteins/digestive enzymes, which are important allergens. Millions of people are affected by allergy and asthma, and their number is growing. In homes, the major triggers of allergy and asthma are the house dust mites Dermatophagoides farinae and D. pteronyssinus, and a clear understanding of the development of diseases caused by these mites is needed. The major sources of mite allergens are their feces, which are deposited in the environment and are easily inhaled as part of aeroplankton. However, descriptions of and comparisons between the major fecal allergens of these two mites are lacking. This study shows that similar group 1 (cysteine protease), 2 (NPC2 family), 6 (chymotrypsin) and 15 (chitinase-like) allergens are present in the feces of these two mite species, as determined by 2D-E mapping, whereas group 3 (trypsin) and 9 (collagenolytic protease) allergens in the feces of the two species are different. The results provide unique MS/MS mapped fingerprints of mite species-specific isoforms in feces. The presence of ubiquitin in mite feces suggests that these proteins participate in the post-translational modification of fecal proteins. The findings are essential for understanding differences between D. farinae and D. pteronyssinus with respect to immunoreactivity, protease activation mechanisms, association with microbes, and food utilization. Copyright © 2017 Elsevier B.V. All rights reserved.

Use of Spatial Sampling and Microbial Source-Tracking Tools for Understanding Fecal Contamination at Two Lake Erie Beaches

USGS Publications Warehouse

Francy, Donna S.; Bertke, Erin E.; Finnegan, Dennis P.; Kephart, Christopher M.; Sheets, Rodney A.; Rhoades, John; Stumpe, Lester

2006-01-01

Source-tracking tools were used to identify potential sources of fecal contamination at two Lake Erie bathing beaches: an urban beach (Edgewater in Cleveland, Ohio) and a beach in a small city (Lakeshore in Ashtabula, Ohio). These tools included identifying spatial patterns of Escherichia coli (E. coli) concentrations in each area, determining weather patterns that caused elevated E. coli, and applying microbial source tracking (MST) techniques to specific sites. Three MST methods were used during this study: multiple antibiotic resistance (MAR) indexing of E. coli isolates and the presence of human-specific genetic markers within two types of bacteria, the genus Bacteroides and the species Enterococcus faecium. At Edgewater, sampling for E. coli was done during 2003-05 at bathing-area sites, at nearshore lake sites, and in shallow ground water in foreshore and backshore areas. Spatial sampling at nearshore lake sites showed that fecal contamination was most likely of local origin; E. coli concentrations near the mouths of rivers and outfalls remote to the beach were elevated (greater than 235 colony-forming units per 100 milliliters (CFU/100 mL)) but decreased along transport pathways to the beach. In addition, E. coli concentrations were generally highest in bathing-area samples collected at 1- and 2-foot water depths, midrange at 3-foot depths, and lowest in nearshore lake samples typically collected 150 feet from the shoreline. Elevated E. coli concentrations at bathing-area sites were generally associated with increased wave heights and rainfall, but not always. E. coli concentrations were often elevated in shallow ground-water samples, especially in samples collected less than 10 feet from the edge of water (near foreshore area). The interaction of shallow ground water and waves may be a mechanism of E. coli storage and accumulation in foreshore sands. Infiltration of bird feces through sand with surface water from rainfall and high waves may be concentrating E. coli in shallow ground water in foreshore and backshore sands. At Lakeshore, sampling for E. coli was done at bathing-area, nearshore lake, and parking-lot sites during 2004-05. Low concentrations of E. coli at nearshore lake sites furthest from the shoreline indicated that fecal contamination was most likely of local origin. High concentrations of E. coli in water and bed sediments at several nearshore lake sites showed that contamination was emanating from several points along the shoreline during wet and dry weather, including the boat ramp, an area near the pond drainage, and parking-lot sediments. Physical evidence confirmed that runoff from the parking lot leads to degradation of water quality at the beach. MST samples were collected to help interpret spatial findings and determine whether sources of fecal contamination were from wastewater or bird feces and if a human-specific marker was present. MAR indices were useful in distinguishing between bird feces and wastewater sources because they were about 10 times higher in the latter. The results from MAR indices agreed with results from the two human-specific markers in some but not all of the samples tested. Bacteroides and enterococci human-specific markers were found on one day at Edgewater and two days at Lakeshore. On three days at Edgewater and two days at Lakeshore, the MAR index indicated a mixed source, but neither marker was found in bathing-water samples; this may be because bacterial indicator concentrations were too low to detect a marker. Multiple tools are needed to help identify sources of fecal contamination at coastal beaches. Spatial sampling identified patterns in E. coli concentrations and yielded information on the physical pathways of contamination. MST methods provided information on whether the source was likely of human or nonhuman origin only; however, MST did not provide information on the pathways of contamination.

Detection of Bovine and Porcine Adenoviruses for Tracing the Source of Fecal Contamination

PubMed Central

Maluquer de Motes, Carlos; Clemente-Casares, Pilar; Hundesa, Ayalkibet; Martín, Margarita; Girones, Rosina

2004-01-01

In this study, a molecular procedure for the detection of adenoviruses of animal origin was developed to evaluate the level of excretion of these viruses by swine and cattle and to design a test to facilitate the tracing of specific sources of environmental viral contamination. Two sets of oligonucleotides were designed, one to detect porcine adenoviruses and the other to detect bovine and ovine adenoviruses. The specificity of the assays was assessed in 31 fecal samples and 12 sewage samples that were collected monthly during a 1-year period. The data also provided information on the environmental prevalence of animal adenoviruses. Porcine adenoviruses were detected in 17 of 24 (70%) pools of swine samples studied, with most isolates being closely related to serotype 3. Bovine adenoviruses were present in 6 of 8 (75%) pools studied, with strains belonging to the genera Mastadenovirus and Atadenovirus and being similar to bovine adenoviruses of types 2, 4, and 7. These sets of primers produced negative results in nested PCR assays when human adenovirus controls and urban-sewage samples were tested. Likewise, the sets of primers previously designed for detection of human adenovirus also produced negative results with animal adenoviruses. These results indicate the importance of further studies to evaluate the usefulness of these tests to trace the source of fecal contamination in water and food and for environmental studies. PMID:15006765

Gastroenteritis outbreak caused by waterborne norovirus at a New Zealand ski resort.

PubMed

Hewitt, Joanne; Bell, Derek; Simmons, Greg C; Rivera-Aban, Malet; Wolf, Sandro; Greening, Gail E

2007-12-01

In July 2006, public health services investigated an outbreak of acute gastroenteritis among staff and visitors of a popular ski resort in southern New Zealand. The source of the outbreak was a drinking water supply contaminated by human sewage. The virological component of the investigation played a major role in confirming the source of the outbreak. Drinking water, source stream water, and 31 fecal specimens from gastroenteritis outbreak cases were analyzed for the presence of norovirus (NoV). Water samples were concentrated by ultrafiltration, and real-time reverse transcription-PCR (RT-PCR) was used for rapid detection of NoV from both water and fecal samples. The implicated NoV strain was further characterized by DNA sequencing. NoV genogroup GI/5 was identified in water samples and linked case fecal specimens, providing clear evidence of the predominant pathogen and route of exposure. A retrospective cohort study demonstrated that staff who consumed drinking water from the resort supply were twice as likely to have gastroenteritis than those who did not. This is the first time that an outbreak of gastroenteritis in New Zealand has been conclusively linked to NoV detected in a community water supply. To our knowledge, this is the first report of the use of ultrafiltration combined with quantitative real-time RT-PCR and DNA sequencing for investigation of a waterborne NoV outbreak.

Detection of bovine and porcine adenoviruses for tracing the source of fecal contamination.

PubMed

Maluquer de Motes, Carlos; Clemente-Casares, Pilar; Hundesa, Ayalkibet; Martín, Margarita; Girones, Rosina

2004-03-01

In this study, a molecular procedure for the detection of adenoviruses of animal origin was developed to evaluate the level of excretion of these viruses by swine and cattle and to design a test to facilitate the tracing of specific sources of environmental viral contamination. Two sets of oligonucleotides were designed, one to detect porcine adenoviruses and the other to detect bovine and ovine adenoviruses. The specificity of the assays was assessed in 31 fecal samples and 12 sewage samples that were collected monthly during a 1-year period. The data also provided information on the environmental prevalence of animal adenoviruses. Porcine adenoviruses were detected in 17 of 24 (70%) pools of swine samples studied, with most isolates being closely related to serotype 3. Bovine adenoviruses were present in 6 of 8 (75%) pools studied, with strains belonging to the genera Mastadenovirus and Atadenovirus and being similar to bovine adenoviruses of types 2, 4, and 7. These sets of primers produced negative results in nested PCR assays when human adenovirus controls and urban-sewage samples were tested. Likewise, the sets of primers previously designed for detection of human adenovirus also produced negative results with animal adenoviruses. These results indicate the importance of further studies to evaluate the usefulness of these tests to trace the source of fecal contamination in water and food and for environmental studies.

Prevalence of Diarrheagenic Escherichia coli in Foods and Fecal Specimens Obtained from Cattle, Pigs, Chickens, Asymptomatic Carriers, and Patients in Osaka and Hyogo, Japan.

PubMed

Wang, Lili; Zhang, Shaobo; Zheng, Dongming; Fujihara, Sami; Wakabayashi, Akiyo; Okahata, Kazuyuki; Suzuki, Masakazu; Saeki, Atsunori; Nakamura, Hiromi; Hara-Kudo, Yukiko; Kage-Nakadai, Eriko; Nishikawa, Yoshikazu

2017-07-24

The source and routes of diarrheagenic Escherichia coli (DEC) remain poorly understood. To investigate the involvement of domestic animals in the dissemination of DEC, the prevalence of DEC in foods and fecal specimens from cattle, pigs, chickens, healthy carriers, and patients in Osaka and Hyogo, Japan was investigated using a multiplex real-time Polymerase Chain Reaction assay. The most abundant virulence genes were astA and eae, which had a prevalence 46.8% and 27.4%, respectively. Additionally, stx1 (26.6%) and stx2 (45.9%) were prevalent in cattle feces, while est (8.5%) and elt (7.6%) were prevalent in pig feces. afaB was the second-most prevalent gene in patients and healthy carriers, and it had detection rates of 5.1% and 8.1%, respectively. In contrast, afaB was not detected in animal feces or foods, except for three porcine fecal samples. The aggR gene was more prevalent in humans than in foods or animal feces. Both Shiga toxin-producing E. coli and atypical enteropathogenic E. coli carried by cattle may be sources for diarrheal diseases in humans. Pigs may be a source for human enterotoxigenic E. coli infections, whereas humans are expected to be the reservoir for diffusely adhering E. coli, enteroaggregative E. coli, and enteroinvasive E. coli.

Using enteric pathogens to assess sources of fecal contamination in the silurian dolomite aquifer: preliminary results

USDA-ARS?s Scientific Manuscript database

The fractured Silurian dolomite aquifer is an important, but vulnerable, source of drinking water in northeast Wisconsin. Areas underlain by the Silurian dolomite aquifer are extremely vulnerable to groundwater contamination from various land-use activities, especially the disposal of human wastewat...

Relationships and trends of E. Coli, human-associated bacteroides, and pathogens in the Proctor Creek watershed (GWRC 2017)

EPA Science Inventory

Urban surface waters can be impacted by anthropogenic sources such as impervious surfaces, sani-tary and storm sewers, and failing infrastructure. Fecal indicator bacteria (FIB) and microbial source tracking (MST) markers are common gauges of stream water qual-ity, however, litt...

Process Relationships for Evaluating the Role of Light-induced Inactivation of Enterococci at Selected Beaches and Nearby Tributaries of the Great Lakes

EPA Science Inventory

One approach to predictive modeling of biological contamination of recreational waters and drinking water sources involves applying process-based models that consider microbial sources, hydrodynamic transport, and microbial fate. Fecal indicator bacteria such as enterococci have ...

Performance of forty-one microbial source tracking methods: A twenty-seven lab evaluation study

EPA Science Inventory

The last decade has seen development of numerous new microbial source tracking (MST) methodologies, but many of these have been tested in just a few laboratories with a limited number of fecal samples. This method evaluation study examined the specificity and sensitivity of 43 ...

Fast Technology Analysis (FTA) Enables Identification of Species and Genotypes of Latent Microsporidia Infections in Healthy Native Cameroonians

PubMed Central

Ndzi, Edward S.; Asonganyi, Tazoacha; Nkinin, Mary Bello; Xiao, Lihua; Didier, Elizabeth S.; Bowers, Lisa C.; Nkinin, Stephenson W.; Kaneshiro, Edna S.

2015-01-01

Several enteric microsporidia species have been detected in humans and other vertebrates and their identifications at the genotype level are currently being elucidated. As advanced methods, reagents, and disposal kits for detecting and identifying pathogens become commercially available, it is important to test them in settings other than in laboratories with “state-of-the-art” equipment and well-trained staff members. In the present study, we sought to detect microsporidia DNA preserved and extracted from FTA (fast technology analysis) cards spotted with human fecal suspensions obtained from Cameroonian volunteers living in the capital city of Yaoundé to preclude the need for employing spore-concentrating protocols. Further, we tested whether amplicon nucleotide sequencing approaches could be used on small aliquots taken from the cards to elucidate the diversity of microsporidia species and strains infecting native residents. Of 196 samples analyzed, 12 (6.1%) were positive for microsporidia DNA; Enterocytozoon bieneusi (Type IV and KIN-1), Encephalitozoon cuniculi, and Encephalitozoon intestinalis were identified. These data demonstrate the utility of the FTA cards in identifying genotypes of microsporidia DNA in human fecal samples that may be applied to field testing for prevalence studies. PMID:26303263

Incidence of the enterococcal surface protein (esp) gene in human and animal fecal sources

USGS Publications Warehouse

Whitman, R.L.; Przybyla-Kelly, K.; Shively, D.A.; Byappanahalli, M.N.

2007-01-01

The occurrence of the enterococcal surface protein (esp) gene in the opportunistic pathogens Enterococcus faecalis and E. faecium is well-documented in clinical research. Recently, the esp gene has been proposed as a marker of human pollution in environmental waters; however, information on its relative incidence in various human and animal fecal sources is limited. We have determined the occurrence of the esp gene in enterococci from human (n = 64) and animal (n = 233) fecal samples by polymerase chain reaction using two primer sets: one presumably specific for E. faecium (espfm) and the other for both E. faecalis and E. faecium (espfs/fm). We believe that this research is the first to explore the use of espfs/fm for the detection of human waste in natural environmental settings. The incidence in human sources was 93.1% espfm and 100% espfs/fm in raw sewage influent; 30% for both espfm and espfs/fm in septic waste; and 0% espfm and 80% espfs/fm in active pit toilets. The overall occurrence of the gene in animal feces was 7.7% (espfs/fm) and 4.7% (espfm); animal types with positive results included dogs (9/43, all espfm), gulls (10/34, espfs/fm; 2/34, espfm), mice (3/22, all espfs/fm), and songbirds (5/55, all espfs/fm). The esp gene was not detected in cat (0/34), deer (0/4), goose (0/18), or raccoon (0/23) feces. The inconsistent occurrence, especially in septic and pit toilet sewage, suggests a low statistical power of discrimination between animal and human sources, which means a large number of replicates should be collected. Both espfm and espfs/fm were common in raw sewage, but neither one efficiently differentiated between animal and other human sources.

Modeling fecal contamination in the Aljezur coastal stream (Portugal)

NASA Astrophysics Data System (ADS)

Rodrigues, Marta; Oliveira, Anabela; Guerreiro, Martha; Fortunato, André Bustorff; Menaia, José; David, Luís Mesquita; Cravo, Alexandra

2011-06-01

This study aims at understanding the fecal contamination behavior in a small coastal stream (Aljezur, Portugal), which has significant economic and ecological values. Like in most small coastal systems, circulation and water renewal in the Aljezur stream exhibit a strong variability due to their dependence on tides, waves, intermittent river flows, and a highly variable morphology. Hence, the problem was approached through a combination of field surveys and the development and application of a hard-coupled three-dimensional hydrodynamic and fecal contamination model. Salinity and temperature results have shown that mixing and transport in the stream are very sensitive to the river flow and wind forcing. The model is able to represent the main patterns and trends observed in Escherichia coli and fecal enterococcus concentrations along the stream, for different environmental and contamination conditions, suggesting die-off rates on the order of 0.50-0.55 day-1. Die-off rate and the representation of the sediment-associated processes were identified as the major remaining sources of uncertainty in the model. Results show that, owing to the processes that occur along the stream, fecal bacteria reach the beaches water in numbers that comply with the European Bathing Waters Directive, even during the summer periods when the upstream concentrations are larger. In particular, results suggest a direct relation between the tidal propagation upstream and the reduction of the fecal bacteria concentrations along the stream that can be relevant for the development of a strategy for the management of the system's water safety.

Left-colon antegrade continence enema (LACE) procedure for fecal incontinence.

PubMed

Churchill, Bernard M; De Ugarte, Daniel A; Atkinson, James B

2003-12-01

Antegrade continence enemas (ACE) are an efficacious therapeutic option for patients with fecal incontinence. The authors review their institution's experience with a variation of the Monti-Malone ACE procedure using the left colon as a source of an intestinal conduit and enema reservoir. From 2000 to 2002, 18 patients with fecal incontinence or intractable constipation underwent left-colon ACE (LACE) procedure. Concomitant Mitrofanoff appendicovesicostomy was performed in 15 patients and bladder augmentation in 9. The majority of patients had neural tube defects. A segment of left colon was tubularized, tunneled into the muscular wall of the distal colon, and exteriorized through the left upper quadrant or midabdomen. Stomal catherization and enema installation were started one month postoperatively. Fifteen patients (83%) achieved fecal continence, 2 remain incontinent of stool, and 1 experienced stomal closure (mean follow-up was 24 +/- 9 months). Two patients had stomal stenosis that required revision. The mean enema volume in patient's achieving continence was 360 +/- 216 mL, and the mean transit time was 18 +/- 12 minutes. LACE is an efficacious procedure for fecal incontinence that can be performed safely at the time of major urologic reconstruction. Administration of enemas into the left colon has several physiologic advantages that result in predictable bowel evacuation.

Water microbiology. Bacterial pathogens and water.

PubMed

Cabral, João P S

2010-10-01

Water is essential to life, but many people do not have access to clean and safe drinking water and many die of waterborne bacterial infections. In this review a general characterization of the most important bacterial diseases transmitted through water-cholera, typhoid fever and bacillary dysentery-is presented, focusing on the biology and ecology of the causal agents and on the diseases' characteristics and their life cycles in the environment. The importance of pathogenic Escherichia coli strains and emerging pathogens in drinking water-transmitted diseases is also briefly discussed. Microbiological water analysis is mainly based on the concept of fecal indicator bacteria. The main bacteria present in human and animal feces (focusing on their behavior in their hosts and in the environment) and the most important fecal indicator bacteria are presented and discussed (focusing on the advantages and limitations of their use as markers). Important sources of bacterial fecal pollution of environmental waters are also briefly indicated. In the last topic it is discussed which indicators of fecal pollution should be used in current drinking water microbiological analysis. It was concluded that safe drinking water for all is one of the major challenges of the 21st century and that microbiological control of drinking water should be the norm everywhere. Routine basic microbiological analysis of drinking water should be carried out by assaying the presence of Escherichia coli by culture methods. Whenever financial resources are available, fecal coliform determinations should be complemented with the quantification of enterococci. More studies are needed in order to check if ammonia is reliable for a preliminary screening for emergency fecal pollution outbreaks. Financial resources should be devoted to a better understanding of the ecology and behavior of human and animal fecal bacteria in environmental waters.

Comparison of the Microbial Community Structures of Untreated Wastewaters from Different Geographic Locales

PubMed Central

Shanks, Orin C.; Newton, Ryan J.; Kelty, Catherine A.; Huse, Susan M.; Sogin, Mitchell L.

2013-01-01

Microbial sewage communities consist of a combination of human fecal microorganisms and nonfecal microorganisms, which may be residents of urban sewer infrastructure or flowthrough originating from gray water or rainwater inputs. Together, these different microorganism sources form an identifiable community structure that may serve as a signature for sewage discharges and as candidates for alternative indicators specific for human fecal pollution. However, the structure and variability of this community across geographic space remains uncharacterized. We used massively parallel 454 pyrosequencing of the V6 region in 16S rRNA genes to profile microbial communities from 13 untreated sewage influent samples collected from a wide range of geographic locations in the United States. We obtained a total of 380,175 high-quality sequences for sequence-based clustering, taxonomic analyses, and profile comparisons. The sewage profile included a discernible core human fecal signature made up of several abundant taxonomic groups within Firmicutes, Bacteroidetes, Actinobacteria, and Proteobacteria. DNA sequences were also classified into fecal, sewage infrastructure (i.e., nonfecal), and transient groups based on data comparisons with fecal samples. Across all sewage samples, an estimated 12.1% of sequences were fecal in origin, while 81.4% were consistently associated with the sewage infrastructure. The composition of feces-derived operational taxonomic units remained congruent across all sewage samples regardless of geographic locale; however, the sewage infrastructure community composition varied among cities, with city latitude best explaining this variation. Together, these results suggest that untreated sewage microbial communities harbor a core group of fecal bacteria across geographically dispersed wastewater sewage lines and that ambient water quality indicators targeting these select core microorganisms may perform well across the United States. PMID:23435885

Frequent detection of a human fecal indicator in the urban ocean: environmental drivers and covariation with enterococci.

PubMed

Jennings, Wiley C; Chern, Eunice C; O'Donohue, Diane; Kellogg, Michael G; Boehm, Alexandria B

2018-03-01

Fecal pollution of surface waters presents a global human health threat. New molecular indicators of fecal pollution have been developed to address shortcomings of traditional culturable fecal indicators. However, there is still little information on their fate and transport in the environment. The present study uses spatially and temporally extensive data on traditional (culturable enterococci, cENT) and molecular (qPCR-enterococci, qENT and human-associated marker, HF183/BacR287) indicator concentrations in marine water surrounding highly-urbanized San Francisco, California, USA to investigate environmental and anthropogenic processes that impact fecal pollution. We constructed multivariable regression models for fecal indicator bacteria at 14 sampling stations. The human marker was detected more frequently in our study than in many other published studies, with detection frequency at some stations as high as 97%. The odds of cENT, qENT, and HF183/BacR287 exceeding health-relevant thresholds were statistically elevated immediately following discharges of partially treated combined sewage, and cENT levels dissipated after approximately 1 day. However, combined sewer discharges were not important predictors of indicator levels typically measured in weekly monitoring samples. Instead, precipitation and solar insolation were important predictors of cENT in weekly samples, while precipitation and water temperature were important predictors of HF183/BacR287 and qENT. The importance of precipitation highlights the significance of untreated storm water as a source of fecal pollution to the urban ocean, even for a city served by a combined sewage system. Sunlight and water temperature likely control persistence of the indicators via photoinactivation and dark decay processes, respectively.

Water Microbiology. Bacterial Pathogens and Water

PubMed Central

Cabral, João P. S.

2010-01-01

Water is essential to life, but many people do not have access to clean and safe drinking water and many die of waterborne bacterial infections. In this review a general characterization of the most important bacterial diseases transmitted through water—cholera, typhoid fever and bacillary dysentery—is presented, focusing on the biology and ecology of the causal agents and on the diseases’ characteristics and their life cycles in the environment. The importance of pathogenic Escherichia coli strains and emerging pathogens in drinking water-transmitted diseases is also briefly discussed. Microbiological water analysis is mainly based on the concept of fecal indicator bacteria. The main bacteria present in human and animal feces (focusing on their behavior in their hosts and in the environment) and the most important fecal indicator bacteria are presented and discussed (focusing on the advantages and limitations of their use as markers). Important sources of bacterial fecal pollution of environmental waters are also briefly indicated. In the last topic it is discussed which indicators of fecal pollution should be used in current drinking water microbiological analysis. It was concluded that safe drinking water for all is one of the major challenges of the 21st century and that microbiological control of drinking water should be the norm everywhere. Routine basic microbiological analysis of drinking water should be carried out by assaying the presence of Escherichia coli by culture methods. Whenever financial resources are available, fecal coliform determinations should be complemented with the quantification of enterococci. More studies are needed in order to check if ammonia is reliable for a preliminary screening for emergency fecal pollution outbreaks. Financial resources should be devoted to a better understanding of the ecology and behavior of human and animal fecal bacteria in environmental waters. PMID:21139855

Genotypic Diversity of Escherichia coli in the Water and Soil of Tropical Watersheds in Hawaii ▿

PubMed Central

Goto, Dustin K.; Yan, Tao

2011-01-01

High levels of Escherichia coli were frequently detected in tropical soils in Hawaii, which present important environmental sources of E. coli to water bodies. This study systematically examined E. coli isolates from water and soil of several watersheds in Hawaii and observed high overall genotypic diversity (35.5% unique genotypes). In the Manoa watershed, fewer than 9.3% of the observed E. coli genotypes in water and 6.6% in soil were shared between different sampling sites, suggesting the lack of dominant fecal sources in the watershed. High temporal variability of E. coli genotypes in soil was also observed, which suggests a dynamic E. coli population corresponding with the frequently observed high concentrations in tropical soils. When E. coli genotypes detected from the same sampling events were compared, limited sharing between the soil and water samples was observed in the majority of comparisons (73.5%). However, several comparisons reported up to 33.3% overlap of E. coli genotypes between soil and water, illustrating the potential for soil-water interactions under favorable environmental conditions. In addition, genotype accumulation curves for E. coli from water and soil indicated that the sampling efforts in the Manoa watershed could not exhaust the overall genotypic diversity. Comparisons of E. coli genotypes from other watersheds on Oahu, Hawaii, identified no apparent grouping according to sampling locations. The results of the present study demonstrate the complexity of using E. coli as a fecal indicator bacterium in tropical watersheds and highlight the need to differentiate environmental sources of E. coli from fecal sources in water quality monitoring. PMID:21515724

Association of Fecal Indicator Bacteria with Human Viruses and Microbial Source Tracking Markers at Coastal Beaches Impacted by Nonpoint Source Pollution

PubMed Central

McQuaig, Shannon; Griffith, John

2012-01-01

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. PMID:22773625

Coliphages as indicators of enteroviruses.

PubMed Central

Stetler, R E

1984-01-01

Coliphages were monitored in conjunction with indicator bacteria and enteroviruses in a drinking-water plant modified to reduce trihalomethane production. Coliphages could be detected in the source water by direct inoculation, and sufficient coliphages were detected in enterovirus concentrates to permit following the coliphage levels through different water treatment processes. The recovery efficiency by different filter types ranged from 1 to 53%. Statistical analysis of the data indicated that enterovirus isolates were better correlated with coliphages than with total coliforms, fecal coliforms, fecal streptococci, or standard plate count organisms. Coliphages were not detected in finished water. PMID:6093694

Cross-Comparison of Human Wastewater-Associated Molecular Markers in Relation to Fecal Indicator Bacteria and Enteric Viruses in Recreational Beach Waters.

PubMed

Hughes, B; Beale, D J; Dennis, P G; Cook, S; Ahmed, W

2017-04-15

Detection of human wastewater contamination in recreational waters is of critical importance to regulators due to the risks posed to public health. To identify such risks, human wastewater-associated microbial source tracking (MST) markers have been developed. At present, however, a greater understanding of the suitability of these markers for the detection of diluted human wastewater in environmental waters is necessary to predict risk. Here, we compared the process limit of detection (PLOD) and process limit of quantification (PLOQ) of six human wastewater-associated MST markers ( Bacteroides HF183 [HF183], Escherichia coli H8 [EC H8], Methanobrevibacter smithii nifH , human adenovirus [HAdV], human polyomavirus [HPyV], and pepper mild mottle virus [PMMoV]) in relation to a fecal indicator bacterium (FIB), Enterococcus sp. 23S rRNA (ENT 23S), and three enteric viruses (human adenovirus serotypes 40/41 [HAdV 40/41], human norovirus [HNoV], and human enterovirus [EV]) in beach water samples seeded with raw and secondary-treated wastewater. Among the six MST markers tested, HF183 was the most sensitive measure of human fecal pollution and was quantifiable up to dilutions of 10 -6 and 10 -4 for beach water samples seeded with raw and secondary-treated wastewater, respectively. Other markers and enteric viruses were detected at various dilutions (10 -1 to 10 -5 ). These MST markers, FIB, and enteric viruses were then quantified in beach water ( n = 12) and sand samples ( n = 12) from South East Queensland (SEQ), Australia, to estimate the levels of human fecal pollution. Of the 12 sites examined, beach water and sand samples from several sites had quantifiable concentrations of HF183 and PMMoV markers. Overall, our results indicate that while HF183 is the most sensitive measure of human fecal pollution, it should be used in conjunction with a conferring viral marker to avoid overestimating the risk of gastrointestinal illness. IMPORTANCE MST is an effective tool to help utilities and regulators improve recreational water quality around the globe. Human fecal pollution poses significant public health risks compared to animal fecal pollution. Several human wastewater-associated markers have been developed and used for MST field studies. However, a head-to-head comparison in terms of their performance to detect diluted human fecal pollution in recreational water is lacking. In this study, we cross-compared the performance of six human wastewater-associated markers in relation to FIB and enteric viruses in beach water samples seeded with raw and secondary-treated wastewater. The results of this study will provide guidance to regulators and utilities on the appropriate application of MST markers for tracking the sources of human fecal pollution in environmental waters and confer human health risks. Copyright © 2017 American Society for Microbiology.

Cross-Comparison of Human Wastewater-Associated Molecular Markers in Relation to Fecal Indicator Bacteria and Enteric Viruses in Recreational Beach Waters

PubMed Central

Hughes, B.; Beale, D. J.; Dennis, P. G.; Cook, S.

2017-01-01

ABSTRACT Detection of human wastewater contamination in recreational waters is of critical importance to regulators due to the risks posed to public health. To identify such risks, human wastewater-associated microbial source tracking (MST) markers have been developed. At present, however, a greater understanding of the suitability of these markers for the detection of diluted human wastewater in environmental waters is necessary to predict risk. Here, we compared the process limit of detection (PLOD) and process limit of quantification (PLOQ) of six human wastewater-associated MST markers (Bacteroides HF183 [HF183], Escherichia coli H8 [EC H8], Methanobrevibacter smithii nifH, human adenovirus [HAdV], human polyomavirus [HPyV], and pepper mild mottle virus [PMMoV]) in relation to a fecal indicator bacterium (FIB), Enterococcus sp. 23S rRNA (ENT 23S), and three enteric viruses (human adenovirus serotypes 40/41 [HAdV 40/41], human norovirus [HNoV], and human enterovirus [EV]) in beach water samples seeded with raw and secondary-treated wastewater. Among the six MST markers tested, HF183 was the most sensitive measure of human fecal pollution and was quantifiable up to dilutions of 10−6 and 10−4 for beach water samples seeded with raw and secondary-treated wastewater, respectively. Other markers and enteric viruses were detected at various dilutions (10−1 to 10−5). These MST markers, FIB, and enteric viruses were then quantified in beach water (n = 12) and sand samples (n = 12) from South East Queensland (SEQ), Australia, to estimate the levels of human fecal pollution. Of the 12 sites examined, beach water and sand samples from several sites had quantifiable concentrations of HF183 and PMMoV markers. Overall, our results indicate that while HF183 is the most sensitive measure of human fecal pollution, it should be used in conjunction with a conferring viral marker to avoid overestimating the risk of gastrointestinal illness. IMPORTANCE MST is an effective tool to help utilities and regulators improve recreational water quality around the globe. Human fecal pollution poses significant public health risks compared to animal fecal pollution. Several human wastewater-associated markers have been developed and used for MST field studies. However, a head-to-head comparison in terms of their performance to detect diluted human fecal pollution in recreational water is lacking. In this study, we cross-compared the performance of six human wastewater-associated markers in relation to FIB and enteric viruses in beach water samples seeded with raw and secondary-treated wastewater. The results of this study will provide guidance to regulators and utilities on the appropriate application of MST markers for tracking the sources of human fecal pollution in environmental waters and confer human health risks. PMID:28159789

Development of a Real-Time PCR for a Sensitive One-Step Coprodiagnosis Allowing both the Identification of Carnivore Feces and the Detection of Toxocara spp. and Echinococcus multilocularis

PubMed Central

Umhang, Gérald; Poulle, Marie-Lazarine; Millon, Laurence

2016-01-01

Studying the environmental occurrence of parasites of concern for humans and animals based on coprosamples is an expanding field of work in epidemiology and the ecology of health. Detecting and quantifying Toxocara spp. and Echinococcus multilocularis, two predominant zoonotic helminths circulating in European carnivores, in feces may help to better target measures for prevention. A rapid, sensitive, and one-step quantitative PCR (qPCR) allowing detection of E. multilocularis and Toxocara spp. was developed in the present study, combined with a host fecal test based on the identification of three carnivores (red fox, dog, and cat) involved in the life cycles of these parasites. A total of 68 coprosamples were collected from identified specimens from Vulpes vulpes, Canis lupus familiaris, Canis lupus, Felis silvestris catus, Meles meles, Martes foina, and Martes martes. With DNA coprosamples, real-time PCR was performed in duplex with a qPCR inhibitor control specifically designed for this study. All the coprosample host identifications were confirmed by qPCR combined with sequencing, and parasites were detected and confirmed (E. multilocularis in red foxes and Toxocara cati in cats; 16% of samples presented inhibition). By combining parasite detection and quantification, the host fecal test, and a new qPCR inhibitor control, we created a technique with a high sensitivity that may considerably improve environmental studies of pathogens. PMID:26969697

Development of a Real-Time PCR for a Sensitive One-Step Coprodiagnosis Allowing both the Identification of Carnivore Feces and the Detection of Toxocara spp. and Echinococcus multilocularis.

PubMed

Knapp, Jenny; Umhang, Gérald; Poulle, Marie-Lazarine; Millon, Laurence

2016-05-15

Studying the environmental occurrence of parasites of concern for humans and animals based on coprosamples is an expanding field of work in epidemiology and the ecology of health. Detecting and quantifying Toxocara spp. and Echinococcus multilocularis, two predominant zoonotic helminths circulating in European carnivores, in feces may help to better target measures for prevention. A rapid, sensitive, and one-step quantitative PCR (qPCR) allowing detection of E. multilocularis and Toxocara spp. was developed in the present study, combined with a host fecal test based on the identification of three carnivores (red fox, dog, and cat) involved in the life cycles of these parasites. A total of 68 coprosamples were collected from identified specimens from Vulpes vulpes, Canis lupus familiaris, Canis lupus, Felis silvestris catus, Meles meles, Martes foina, and Martes martes With DNA coprosamples, real-time PCR was performed in duplex with a qPCR inhibitor control specifically designed for this study. All the coprosample host identifications were confirmed by qPCR combined with sequencing, and parasites were detected and confirmed (E. multilocularis in red foxes and Toxocara cati in cats; 16% of samples presented inhibition). By combining parasite detection and quantification, the host fecal test, and a new qPCR inhibitor control, we created a technique with a high sensitivity that may considerably improve environmental studies of pathogens. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

Molecular identification of Enterocytozoon bieneusi, Cryptosporidium, and Giardia in Brazilian captive birds.

PubMed

da Cunha, Maria Júlia Rodrigues; Cury, Márcia Cristina; Santín, Mónica

2017-02-01

A total of 85 fecal samples from captive birds collected from October 2013 to September 2014 in Uberlândia and Belo Horizonte in the state of Minas Gerais (Brazil) were evaluated for the presence of Enterocytozoon bieneusi, Cryptosporidium, and Giardia by PCR. Of these, three birds were found positive for E. bieneusi (3.5%), two for Cryptosporidium (2.3%), and one for Giardia (1.2%). Two genotypes of E. bieneusi were detected by nucleotide sequence analysis of the ITS region, genotypes D and Peru 6 in a swan goose and in two rock pigeons, respectively. For Cryptosporidium and Giardia, nucleotide sequence analysis of the SSU rRNA identified Cryptosporidium baileyi and Duck genotype in a swan goose and a mandarin duck, respectively, and Giardia duodenalis assemblage A in a toco toucon. Our results demonstrate that human-pathogenic E. bieneusi genotypes D and Peru6 and G. duodenalis assemblage A are present in captive birds in Brazil, corroborating their potential role as a source of human infection and environmental contamination.

Shigellosis in the Marshall Islands: epidemiologic aspects of an outbreak.

PubMed

Storch, G A; Gunn, R A; Martin, W T; Pollard, R A; Sinclair, S P

1980-05-01

An outbreak of diarrheal illness caused by Shigella flexneri 1b and associated with 11 deaths occurred in the Marshall Islands during June and July 1977. A house-to-house survey on Majuro indicated an attack rate of 6.2%. Neither the survey nor a case-control study uncovered a common source of exposure on Majuro, and it is believed that transmission was mainly person-to-person. Socioeconomic factors, type of water supply, distance to municipal water supply, and type of sanitary facility could not be related to the occurrence of illness, but a poor sanitary rating was associated with increased rate of household transmission. Absence of stool culturing for bacteria and false-positive identifications of amebae in stool specimens led to the outbreak's being attributed to Entamoeba histolytica. Subsequent parasitologic examinations and serologic studies indicated that E. histolytica played no role in the outbreak, and suggested that fecal leukocytes were being mistaken for amebae. Improved bacteriologic capabilities will be important in improving the approach to diarrheal illness in the Marshall Islands.

Sigma: Strain-level inference of genomes from metagenomic analysis for biosurveillance

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ahn, Tae-Hyuk; Chai, Juanjuan; Pan, Chongle

Motivation: Metagenomic sequencing of clinical samples provides a promising technique for direct pathogen detection and characterization in biosurveillance. Taxonomic analysis at the strain level can be used to resolve serotypes of a pathogen in biosurveillance. Sigma was developed for strain-level identification and quantification of pathogens using their reference genomes based on metagenomic analysis. Results: Sigma provides not only accurate strain-level inferences, but also three unique capabilities: (i) Sigma quantifies the statistical uncertainty of its inferences, which includes hypothesis testing of identified genomes and confidence interval estimation of their relative abundances; (ii) Sigma enables strain variant calling by assigning metagenomic readsmore » to their most likely reference genomes; and (iii) Sigma supports parallel computing for fast analysis of large datasets. In conclusion, the algorithm performance was evaluated using simulated mock communities and fecal samples with spike-in pathogen strains. Availability and Implementation: Sigma was implemented in C++ with source codes and binaries freely available at http://sigma.omicsbio.org.« less

Sigma: Strain-level inference of genomes from metagenomic analysis for biosurveillance

DOE PAGES

Ahn, Tae-Hyuk; Chai, Juanjuan; Pan, Chongle

2014-09-29

Motivation: Metagenomic sequencing of clinical samples provides a promising technique for direct pathogen detection and characterization in biosurveillance. Taxonomic analysis at the strain level can be used to resolve serotypes of a pathogen in biosurveillance. Sigma was developed for strain-level identification and quantification of pathogens using their reference genomes based on metagenomic analysis. Results: Sigma provides not only accurate strain-level inferences, but also three unique capabilities: (i) Sigma quantifies the statistical uncertainty of its inferences, which includes hypothesis testing of identified genomes and confidence interval estimation of their relative abundances; (ii) Sigma enables strain variant calling by assigning metagenomic readsmore » to their most likely reference genomes; and (iii) Sigma supports parallel computing for fast analysis of large datasets. In conclusion, the algorithm performance was evaluated using simulated mock communities and fecal samples with spike-in pathogen strains. Availability and Implementation: Sigma was implemented in C++ with source codes and binaries freely available at http://sigma.omicsbio.org.« less

DETERMINING HOT SPOTS OF FECAL CONTAMINATION IN A TROPICAL WATERSHED BY COMBINING LAND-USE INFORMATION AND METEOROLOGICAL DATA WITH SOURCE-SPECIFIC ASSAYS

EPA Science Inventory

Microbial source tracking (MST) assays have been mostly employed in temperate climates. However, their value as monitoring tools in tropical and subtropical regions is unknown since the geographic and temporal stability of the assays has not been extensively tested. The objective...

EVALUATE THE UTILITY OF ENTEROCOCCI AS INDICATORS OF THE SOURCES OF FECAL CONTAMINATION IN IMPAIRED SUBWATERSHEDS THROUGH DNA-BASED MOLECULAR TECHNIQUES

EPA Science Inventory

Microbial source tracking (MST) is based on the assumption that specific strains of bacteria are associated with specific host species. MST methods are attractive because their application on environmental samples could help define the nature of water quality problems in impaire...

DEVELOPMENT OF HOST-SPECIFIC METAGENOMIC MARKERS FOR MICROBIAL SOURCE TRACKING USING A NOVEL METAGENOMIC APPROACH

EPA Science Inventory

Fecal contamination of source water has always been an important issue to the drinking water industry. Improper disposal of animal waste, leaky septic tanks, storm runoff, and the abundance of wildlife in natural water systems can all be responsible for the spread of enteric path...

Examining trends in general fecal indicator bacteria and microbial source tracking genetic markers at non-point source impacted Chicago beaches

EPA Science Inventory

In the Chicago area, treated wastewater and storm water flow through the engineered Chicago River system to the Mississippi River, with the goal to protect Lake Michigan from urban discharges. Therefore, under dry weather conditions, nearby Lake Michigan recreational beaches shou...

Non-invasive identification of protein biomarkers for early pregnancy diagnosis in the cheetah (Acinonyx jubatus).

PubMed

Koester, Diana C; Wildt, David E; Maly, Morgan; Comizzoli, Pierre; Crosier, Adrienne E

2017-01-01

Approximately 80% of cheetahs living in typical zoological collections never reproduce. In more than 60% of breedings, the female is confirmed to ovulate, but parturition fails to occur. It is unknown if these non-pregnant intervals of elevated progesterone (deemed luteal phases) are conception failures or a pregnancy terminating in embryonic/fetal loss. There have been recent advances in metabolic profiling and proteome analyses in many species with mass spectrometry used to identify 'biomarkers' and mechanisms indicative of specific physiological states (including pregnancy). Here, we hypothesized that protein expression in voided cheetah feces varied depending on pregnancy status. We: 1) identified the expansive protein profile present in fecal material of females; and 2) isolated proteins that may be candidates playing a role in early pregnancy establishment and diagnosis. Five hundred and seventy unique proteins were discovered among samples from pregnant (n = 8), non-pregnant, luteal phase (n = 5), and non-ovulatory control (n = 5) cheetahs. Four protein candidates were isolated that were significantly up-regulated and two were down-regulated in samples from pregnant compared to non-pregnant or control counterparts. One up-regulated candidate, immunoglobulin J chain (IGJ; an important component of the secretory immune system) was detected using a commercially available antibody via immunoblotting. Findings revealed that increased IGJ abundance could be used to detect pregnancy successfully in >80% of 23 assessed females within 4 weeks after mating. The discovery of a novel fecal pregnancy marker improves the ability to determine reproductive, especially gestational, status in cheetahs managed in an ex situ insurance and source population.

Non-invasive identification of protein biomarkers for early pregnancy diagnosis in the cheetah (Acinonyx jubatus)

PubMed Central

Wildt, David E.; Maly, Morgan; Comizzoli, Pierre; Crosier, Adrienne E.

2017-01-01

Approximately 80% of cheetahs living in typical zoological collections never reproduce. In more than 60% of breedings, the female is confirmed to ovulate, but parturition fails to occur. It is unknown if these non-pregnant intervals of elevated progesterone (deemed luteal phases) are conception failures or a pregnancy terminating in embryonic/fetal loss. There have been recent advances in metabolic profiling and proteome analyses in many species with mass spectrometry used to identify ‘biomarkers’ and mechanisms indicative of specific physiological states (including pregnancy). Here, we hypothesized that protein expression in voided cheetah feces varied depending on pregnancy status. We: 1) identified the expansive protein profile present in fecal material of females; and 2) isolated proteins that may be candidates playing a role in early pregnancy establishment and diagnosis. Five hundred and seventy unique proteins were discovered among samples from pregnant (n = 8), non-pregnant, luteal phase (n = 5), and non-ovulatory control (n = 5) cheetahs. Four protein candidates were isolated that were significantly up-regulated and two were down-regulated in samples from pregnant compared to non-pregnant or control counterparts. One up-regulated candidate, immunoglobulin J chain (IGJ; an important component of the secretory immune system) was detected using a commercially available antibody via immunoblotting. Findings revealed that increased IGJ abundance could be used to detect pregnancy successfully in >80% of 23 assessed females within 4 weeks after mating. The discovery of a novel fecal pregnancy marker improves the ability to determine reproductive, especially gestational, status in cheetahs managed in an ex situ insurance and source population. PMID:29236714

Hypersaline groundwater genesis assessment through a multidisciplinary approach: the case of Pozzo del Sale Spring (southern Italy)

NASA Astrophysics Data System (ADS)

Celico, Fulvio; Capuano, Paolo; de Felice, Vincenzo; Naclerio, Gino

2008-11-01

A tool, based on a multidisciplinary field investigation approach for studying the characteristics of a hypersaline spring, was developed and its effectiveness tested on a spring in southern Italy; a preliminary model of the aquifer system at medium and local scale was derived. Hydrologic measurements, vertical electric soundings, and chemical and isotopic (δ18O, δ2H, 3H) analyses were undertaken, along with microbiological analyses and species identification. These demonstrate the coexistence of hypersaline and fresh water, generating a significant diversification of the groundwater hydrochemical signature. The isotopic signature shows that both types of water have a meteoric origin. Microbial contamination of fecal origin indicates the mixing of hyper- and low- saline water related to local infiltration. The hypersaline groundwater flows in confined horizons within a sequence that is mainly of fractured clays. These horizons are probably concentrated where well-developed fracture network and dissolution openings within evaporitic rocks enhance fluid flow. In a wider context, this study determines that microbiological pollution of saline groundwater may not be detected if using nonhalophilic bacterial indicators such as fecal coliforms. Fecal enterococci are better indicators, due to their higher halotolerance.

Development of an Enhanced Metaproteomic Approach for Deepening the Microbiome Characterization of the Human Infant Gut

DOE Office of Scientific and Technical Information (OSTI.GOV)

Xiong, Weili; Richard J. Giannone; Morowitz, Michael J.

The early-life microbiota establishment in the human infant gut is highly variable and plays a crucial role in host nutrients and immunity maturation. While high-performance mass spectrometry (MS)-based metaproteomics is a powerful method for the functional characterization of complex microbial communities, the construction of comprehensive metaproteomic information in human fecal samples is inhibited by the presence of abundant human proteins. To alleviate this restriction, we have designed a novel metaproteomic strategy based on Double Filtering (DF) to enhance microbial protein characterization in complex fecal samples from healthy premature infants. We improved the overall depth of infant gut proteome measurement, withmore » an increase in the number of identified low abundance proteins, and observed greater than twofold improvement in metrics for microbial protein identifications and quantifications with a relatively high rank correlation to control. We further showed the substantial enhancement of this approach for extensively interpreting microbial functional categories between infants by affording more detailed and confident identified categories. This approach provided an avenue for in-depth measurement in the microbial component of infant fecal samples and thus comprehensive characterization of infant gut microbiome functionality.« less

Development of an Enhanced Metaproteomic Approach for Deepening the Microbiome Characterization of the Human Infant Gut

DOE PAGES

Xiong, Weili; Richard J. Giannone; Morowitz, Michael J.; ...

2014-10-28

The early-life microbiota establishment in the human infant gut is highly variable and plays a crucial role in host nutrients and immunity maturation. While high-performance mass spectrometry (MS)-based metaproteomics is a powerful method for the functional characterization of complex microbial communities, the construction of comprehensive metaproteomic information in human fecal samples is inhibited by the presence of abundant human proteins. To alleviate this restriction, we have designed a novel metaproteomic strategy based on Double Filtering (DF) to enhance microbial protein characterization in complex fecal samples from healthy premature infants. We improved the overall depth of infant gut proteome measurement, withmore » an increase in the number of identified low abundance proteins, and observed greater than twofold improvement in metrics for microbial protein identifications and quantifications with a relatively high rank correlation to control. We further showed the substantial enhancement of this approach for extensively interpreting microbial functional categories between infants by affording more detailed and confident identified categories. This approach provided an avenue for in-depth measurement in the microbial component of infant fecal samples and thus comprehensive characterization of infant gut microbiome functionality.« less

An uncooked vegan diet shifts the profile of human fecal microflora: computerized analysis of direct stool sample gas-liquid chromatography profiles of bacterial cellular fatty acids.

PubMed Central

Peltonen, R; Ling, W H; Hänninen, O; Eerola, E

1992-01-01

The effect of an uncooked extreme vegan diet on fecal microflora was studied by direct stool sample gas-liquid chromatography (GLC) of bacterial cellular fatty acids and by quantitative bacterial culture by using classical microbiological techniques of isolation, identification, and enumeration of different bacterial species. Eighteen volunteers were divided randomly into two groups. The test group received an uncooked vegan diet for 1 month and a conventional diet of mixed Western type for the other month of the study. The control group consumed a conventional diet throughout the study period. Stool samples were collected. Bacterial cellular fatty acids were extracted directly from the stool samples and measured by GLC. Computerized analysis of the resulting fatty acid profiles was performed. Such a profile represents all bacterial cellular fatty acids in a sample and thus reflects its microflora and can be used to detect changes, differences, or similarities of bacterial flora between individual samples or sample groups. GLC profiles changed significantly in the test group after the induction and discontinuation of the vegan diet but not in the control group at any time, whereas quantitative bacterial culture did not detect any significant change in fecal bacteriology in either of the groups. The results suggest that an uncooked extreme vegan diet alters the fecal bacterial flora significantly when it is measured by direct stool sample GLC of bacterial fatty acids. PMID:1482187

Effect of Dietary Starch Source and Concentration on Equine Fecal Microbiota

PubMed Central

Harlow, Brittany E.; Lawrence, Laurie M.; Hayes, Susan H.; Crum, Andrea; Flythe, Michael D.

2016-01-01

Starch from corn is less susceptible to equine small intestinal digestion than starch from oats, and starch that reaches the hindgut can be utilized by the microbiota. The objective of the current study was to examine the effects of starch source on equine fecal microbiota. Thirty horses were assigned to treatments: control (hay only), HC (high corn), HO (high oats), LC (low corn), LO (low oats), and LW (low pelleted wheat middlings). Horses received an all-forage diet (2 wk; d -14 to d -1) before the treatment diets (2 wk; d 1 to 14). Starch was introduced gradually so that horses received 50% of the assigned starch amount (high = 2 g starch/kg BW; low = 1 g starch/kg BW) by d 4 and 100% by d 11. Fecal samples were obtained at the end of the forage-only period (S0; d -2), and on d 6 (S1) and d 13 (S2) of the treatment period. Cellulolytics, lactobacilli, Group D Gram-positive cocci (GPC), lactate-utilizers and amylolytics were enumerated. Enumeration data were log transformed and analyzed by repeated measures ANOVA. There were sample day × treatment interactions (P < 0.0001) for all bacteria enumerated. Enumerations from control horses did not change during the sampling period (P > 0.05). All treatments except LO resulted in increased amylolytics and decreased cellulolytics, but the changes were larger in horses fed corn and wheat middlings (P < 0.05). Feeding oats resulted in increased lactobacilli and decreased GPC (P < 0.05), while corn had the opposite effects. LW had increased lactobacilli and GPC (P < 0.05). The predominant amylolytic isolates from HC, LC and LW on S2 were identified by 16S RNA gene sequencing as Enterococcus faecalis, but other species were found in oat fed horses. These results demonstrate that starch source can have a differential effect on the equine fecal microbiota. PMID:27128793

Evaluation of new gyrB-based real-time PCR system for the detection of B. fragilis as an indicator of human-specific fecal contamination.

PubMed

Lee, Chang Soo; Lee, Jiyoung

2010-09-01

A rapid and specific gyrB-based real-time PCR system has been developed for detecting Bacteroides fragilis as a human-specific marker of fecal contamination. Its specificity and sensitivity was evaluated by comparison with other 16S rRNA gene-based primers using closely related Bacteroides and Prevotella. Many studies have used 16S rRNA gene-based method targeting Bacteroides because this genus is relatively abundant in human feces and is useful for microbial source tracking. However, 16S rRNA gene-based primers are evolutionarily too conserved among taxa to discriminate between human-specific species of Bacteroides and other closely related genera, such as Prevotella. Recently, one of the housekeeping genes, gyrB, has been used as an alternative target in multilocus sequence analysis (MLSA) to provide greater phylogenetic resolution. In this study, a new B. fragilis-specific primer set (Bf904F/Bf958R) was designed by alignments of 322 gyrB genes and was compared with the performance of the 16S rRNA gene-based primers in the presence of B. fragilis, Bacteroides ovatus and Prevotella melaninogenica. Amplicons were sequenced and a phylogenetic tree was constructed to confirm the specificity of the primers to B. fragilis. The gyrB-based primers successfully discriminated B. fragilis from B. ovatus and P. melaninogenica. Real-time PCR results showed that the gyrB primer set had a comparable sensitivity in the detection of B. fragilis when compared with the 16S rRNA primer set. The host-specificity of our gyrB-based primer set was validated with human, pig, cow, and dog fecal samples. The gyrB primer system had superior human-specificity. The gyrB-based system can rapidly detect human-specific fecal source and can be used for improved source tracking of human contamination. (c) 2010 Elsevier B.V. All rights reserved.

Evaluation of Escherichia coli isolates from healthy chickens to determine their potential risk to poultry and human health.

PubMed

Stromberg, Zachary R; Johnson, James R; Fairbrother, John M; Kilbourne, Jacquelyn; Van Goor, Angelica; Curtiss, Roy; Mellata, Melha

2017-01-01

Extraintestinal pathogenic Escherichia coli (ExPEC) strains are important pathogens that cause diverse diseases in humans and poultry. Some E. coli isolates from chicken feces contain ExPEC-associated virulence genes, so appear potentially pathogenic; they conceivably could be transmitted to humans through handling and/or consumption of contaminated meat. However, the actual extraintestinal virulence potential of chicken-source fecal E. coli is poorly understood. Here, we assessed whether fecal E. coli isolates from healthy production chickens could cause diseases in a chicken model of avian colibacillosis and three rodent models of ExPEC-associated human infections. From 304 E. coli isolates from chicken fecal samples, 175 E. coli isolates were screened by PCR for virulence genes associated with human-source ExPEC or avian pathogenic E. coli (APEC), an ExPEC subset that causes extraintestinal infections in poultry. Selected isolates genetically identified as ExPEC and non-ExPEC isolates were assessed in vitro for virulence-associated phenotypes, and in vivo for disease-causing ability in animal models of colibacillosis, sepsis, meningitis, and urinary tract infection. Among the study isolates, 13% (40/304) were identified as ExPEC; the majority of these were classified as APEC and uropathogenic E. coli, but none as neonatal meningitis E. coli. Multiple chicken-source fecal ExPEC isolates resembled avian and human clinical ExPEC isolates in causing one or more ExPEC-associated illnesses in experimental animal infection models. Additionally, some isolates that were classified as non-ExPEC were able to cause ExPEC-associated illnesses in animal models, and thus future studies are needed to elucidate their mechanisms of virulence. These findings show that E. coli isolates from chicken feces contain ExPEC-associated genes, exhibit ExPEC-associated in vitro phenotypes, and can cause ExPEC-associated infections in animal models, and thus may pose a health threat to poultry and consumers.

Evaluation of Escherichia coli isolates from healthy chickens to determine their potential risk to poultry and human health

PubMed Central

Johnson, James R.; Fairbrother, John M.; Kilbourne, Jacquelyn; Van Goor, Angelica; Curtiss, Roy; Mellata, Melha

2017-01-01

Extraintestinal pathogenic Escherichia coli (ExPEC) strains are important pathogens that cause diverse diseases in humans and poultry. Some E. coli isolates from chicken feces contain ExPEC-associated virulence genes, so appear potentially pathogenic; they conceivably could be transmitted to humans through handling and/or consumption of contaminated meat. However, the actual extraintestinal virulence potential of chicken-source fecal E. coli is poorly understood. Here, we assessed whether fecal E. coli isolates from healthy production chickens could cause diseases in a chicken model of avian colibacillosis and three rodent models of ExPEC-associated human infections. From 304 E. coli isolates from chicken fecal samples, 175 E. coli isolates were screened by PCR for virulence genes associated with human-source ExPEC or avian pathogenic E. coli (APEC), an ExPEC subset that causes extraintestinal infections in poultry. Selected isolates genetically identified as ExPEC and non-ExPEC isolates were assessed in vitro for virulence-associated phenotypes, and in vivo for disease-causing ability in animal models of colibacillosis, sepsis, meningitis, and urinary tract infection. Among the study isolates, 13% (40/304) were identified as ExPEC; the majority of these were classified as APEC and uropathogenic E. coli, but none as neonatal meningitis E. coli. Multiple chicken-source fecal ExPEC isolates resembled avian and human clinical ExPEC isolates in causing one or more ExPEC-associated illnesses in experimental animal infection models. Additionally, some isolates that were classified as non-ExPEC were able to cause ExPEC-associated illnesses in animal models, and thus future studies are needed to elucidate their mechanisms of virulence. These findings show that E. coli isolates from chicken feces contain ExPEC-associated genes, exhibit ExPEC-associated in vitro phenotypes, and can cause ExPEC-associated infections in animal models, and thus may pose a health threat to poultry and consumers. PMID:28671990

Enterococcus phages as potential tool for identifying sewage inputs in the Great Lakes region

USGS Publications Warehouse

Vijayavel, K.; Byappanahalli, Muruleedhara N.; Whitman, Richard L.; Ebdon, J.; Taylor, H.; Kashian, D.R.

2014-01-01

Bacteriophages are viruses living in bacteria that can be used as a tool to detect fecal contamination in surface waters around the world. However, the lack of a universal host strain makes them unsuitable for tracking fecal sources. We evaluated the suitability of two newly isolated Enterococcus host strains (ENT-49 and ENT-55) capable for identifying sewage contamination in impacted waters by targeting phages specific to these hosts. Both host strains were isolated from wastewater samples and identified as E. faecium by 16S rRNA gene sequencing. Occurrence of Enterococcus phages was evaluated in sewage samples (n = 15) from five wastewater treatment plants and in fecal samples from twenty-two species of wild and domesticated animals (individual samples; n = 22). Levels of Enterococcus phages, F + coliphages, Escherichia coli and enterococci were examined from four rivers, four beaches, and three harbors. Enterococcus phages enumeration was at similar levels (Mean = 6.72 Log PFU/100 mL) to F + coliphages in all wastewater samples, but were absent from all non-human fecal sources tested. The phages infecting Enterococcus spp. and F + coliphages were not detected in the river samples (detection threshold < 10 PFU/100 mL), but were present in the beach and harbor samples (range = 1.83 to 2.86 Log PFU/100 mL). Slightly higher concentrations (range = 3.22 to 3.69 Log MPN/100 mL) of E. coli and enterococci when compared to F + coliphages and Enterococcus phages, were observed in the river, beach and harbor samples. Our findings suggest that the bacteriophages associated with these particular Enterococcus host strains offer potentially sensitive and human-source specific indicators of enteric pathogen risk.

Genotypic Characterization of Enterococci Collected from a Coastal Area Using PFGE

NASA Astrophysics Data System (ADS)

Furukawa, Takashi; Yoshida, Terutoyo; Suzuki, Yoshihiro

Development of microbial source tracking (MST) is crucial to ensure public health and bacteriological safety in coastal areas. In this study, pulsed-field gel electrophoresis (PFGE) , utilized for pollution source tracking of hospital infections, was applied to the aquatic environment as a MST method. Enterococcus faecium and Entecococcus faecalis, fecal indicator bacterias, were isolated and identified from a port area, and the genotype was analyzed by PFGE. A total of 230 enterococcal strains were isolated by PFGE, and forty strains were identified as E. faecium using both PCR and the Api 20 Strep test. Dendrogram analysis of the PFGE types revealed that all the 40 E. faecium strains can be classified separately into 7 different groups at a 0.7 similarity level. The genotypic characterization of E. faecium from the coastal port area revealed diverse types. It is suggested that PFGE can be used to differentiate and characterize fecal pollution indicating bacteria by applying it to the aquatic environment, as well as provide detailed information to specify the pollution source.

Density estimation in tiger populations: combining information for strong inference

USGS Publications Warehouse

Gopalaswamy, Arjun M.; Royle, J. Andrew; Delampady, Mohan; Nichols, James D.; Karanth, K. Ullas; Macdonald, David W.

2012-01-01

A productive way forward in studies of animal populations is to efficiently make use of all the information available, either as raw data or as published sources, on critical parameters of interest. In this study, we demonstrate two approaches to the use of multiple sources of information on a parameter of fundamental interest to ecologists: animal density. The first approach produces estimates simultaneously from two different sources of data. The second approach was developed for situations in which initial data collection and analysis are followed up by subsequent data collection and prior knowledge is updated with new data using a stepwise process. Both approaches are used to estimate density of a rare and elusive predator, the tiger, by combining photographic and fecal DNA spatial capture–recapture data. The model, which combined information, provided the most precise estimate of density (8.5 ± 1.95 tigers/100 km2 [posterior mean ± SD]) relative to a model that utilized only one data source (photographic, 12.02 ± 3.02 tigers/100 km2 and fecal DNA, 6.65 ± 2.37 tigers/100 km2). Our study demonstrates that, by accounting for multiple sources of available information, estimates of animal density can be significantly improved.

Density estimation in tiger populations: combining information for strong inference.

PubMed

Gopalaswamy, Arjun M; Royle, J Andrew; Delampady, Mohan; Nichols, James D; Karanth, K Ullas; Macdonald, David W

2012-07-01

A productive way forward in studies of animal populations is to efficiently make use of all the information available, either as raw data or as published sources, on critical parameters of interest. In this study, we demonstrate two approaches to the use of multiple sources of information on a parameter of fundamental interest to ecologists: animal density. The first approach produces estimates simultaneously from two different sources of data. The second approach was developed for situations in which initial data collection and analysis are followed up by subsequent data collection and prior knowledge is updated with new data using a stepwise process. Both approaches are used to estimate density of a rare and elusive predator, the tiger, by combining photographic and fecal DNA spatial capture-recapture data. The model, which combined information, provided the most precise estimate of density (8.5 +/- 1.95 tigers/100 km2 [posterior mean +/- SD]) relative to a model that utilized only one data source (photographic, 12.02 +/- 3.02 tigers/100 km2 and fecal DNA, 6.65 +/- 2.37 tigers/100 km2). Our study demonstrates that, by accounting for multiple sources of available information, estimates of animal density can be significantly improved.

The Ecological Dynamics of Fecal Contamination and Salmonella Typhi and Salmonella Paratyphi A in Municipal Kathmandu Drinking Water

PubMed Central

Walker, Alan W.; Thompson, Corinne N.; Torres, Andres; Dongol, Sabina; Tran Vu Thieu, Nga; Pham Thanh, Duy; Tran Thi Ngoc, Dung; Voong Vinh, Phat; Singer, Andrew C.; Parkhill, Julian; Thwaites, Guy; Basnyat, Buddha; Ferguson, Neil; Baker, Stephen

2016-01-01

One of the UN sustainable development goals is to achieve universal access to safe and affordable drinking water by 2030. It is locations like Kathmandu, Nepal, a densely populated city in South Asia with endemic typhoid fever, where this goal is most pertinent. Aiming to understand the public health implications of water quality in Kathmandu we subjected weekly water samples from 10 sources for one year to a range of chemical and bacteriological analyses. We additionally aimed to detect the etiological agents of typhoid fever and longitudinally assess microbial diversity by 16S rRNA gene surveying. We found that the majority of water sources exhibited chemical and bacterial contamination exceeding WHO guidelines. Further analysis of the chemical and bacterial data indicated site-specific pollution, symptomatic of highly localized fecal contamination. Rainfall was found to be a key driver of this fecal contamination, correlating with nitrates and evidence of S. Typhi and S. Paratyphi A, for which DNA was detectable in 333 (77%) and 303 (70%) of 432 water samples, respectively. 16S rRNA gene surveying outlined a spectrum of fecal bacteria in the contaminated water, forming complex communities again displaying location-specific temporal signatures. Our data signify that the municipal water in Kathmandu is a predominant vehicle for the transmission of S. Typhi and S. Paratyphi A. This study represents the first extensive spatiotemporal investigation of water pollution in an endemic typhoid fever setting and implicates highly localized human waste as the major contributor to poor water quality in the Kathmandu Valley. PMID:26735696