Foreshore sand as a source of Escherichia coli in nearshore water of a Lake Michigan beach

Swimming advisories due to excessive Escherichia coli concentrations are common at 63rd Street Beach, Chicago, Ill. An intensive study was undertaken to characterize the source and fate of E. coli in beach water and sand at the beach. From April through September 2000, water and sand samples were collected daily or twice daily at two depths on three consecutive days per week (water samples, n = 1,747; sand samples, n = 858); hydrometeorological conditions and bird and bather distributions were also recorded. E. coli concentrations in sand and water were significantly correlated, with the highest concentration being found in foreshore sand, followed by those in submerged sediment and water of increasing depth. Gull contributions to E. coli densities in sand and water were most apparent on the day following gull activity in a given area. E. coli recolonized newly placed foreshore sand within 2 weeks. Analysis of variance, correlation, cluster analyses, concentration gradients, temporal-spatial distribution, demographic patterns, and DNA fingerprinting suggest that E. coli may be able to sustain population density in temperate beach sand during summer months without external inputs. This research presents evidence that foreshore beach sand (i) plays a major role in bacterial lake water quality, (ii) is an important non-point source of E. coli to lake water rather than a net sink, (iii) may be environmentally, and perhaps hygienically, problematic, and (iv) is possibly capable of supporting an autochthonous, high density of indicator bacteria for sustained periods, independent of lake, human, or animal input.     

Detection of genetic markers of fecal indicator bacteria in Lake Michigan and determination of their relationship to Escherichia coli densities using standard microbiological methods

Lake Michigan surface waters impacted by fecal pollution were assessed to determine the occurrence of genetic markers for Bacteroides and Escherichia coli. Initial experiments with sewage treatment plant influent demonstrated that total Bacteroides spp. could be detected by PCR in a 25- to 125-fold-higher dilution series than E. coli and human-specific Bacteroides spp., which were both found in similar dilution ranges. The limit of detection for the human-specific genetic marker ranged from 0.2 CFU/100 ml to 82 CFU/100 ml culturable E. coli for four wastewater treatment plants in urban and rural areas. The spatial and temporal distributions of these markers were assessed following major rain events that introduced urban storm water, agricultural runoff, and sewage overflows into Lake Michigan. Bacteroides spp. were detected in all of these samples by PCR, including those with <1 CFU/100 ml E. coli. Human-specific Bacteroides spp. were detected as far as 2 km into Lake Michigan during sewage overflow events, with variable detection 1 to 9 days postoverflow, whereas the cow-specific Bacteroides spp. were detected in only highly contaminated samples near the river outflow. Lake Michigan beaches were also assessed throughout the summer season for the same markers. Bacteroides spp. were detected in all beach samples, including 28 of the 74 samples that did not exceed 235 CFU/100 ml of E. coli. Human-specific Bacteroides spp. were detected at three of the seven beaches; one of the sites demonstrating positive results was sampled during a reported sewage overflow, but E. coli levels were below 235 CFU/100 ml. This study demonstrates the usefulness of non-culture-based microbial-source tracking approaches and the prevalence of these genetic markers in the Great Lakes, including freshwater coastal beaches.     

Genetic relatedness of Escherichia coli isolates in interstitial water from a Lake Huron (Canada) beach

Research was undertaken to characterize Escherichia coli isolates in interstitial water samples of a sandy beach on the southeastern shore of Lake Huron, Ontario, Canada. A survey of the beach area revealed the highest abundance of E. coli in interstitial water of the foreshore beach sand next to the swash zone. Higher concentrations of E. coli (up to 1.6 x 10(6) CFU/100 ml of water) were observed in the interstitial water from the sampling holes on the beach itself compared to lake water and sediment. Repetitive extragenic palindromic PCR (REP-PCR) was used to characterize the genetic diversity of E. coli isolates from interstitial water samples on the beach. E. coli isolates from the same sampling location frequently exhibited the same REP-PCR pattern or were highly similar to each other. In contrast, E. coli isolates from different sampling locations represented populations distinct from each other. This study has identified a unique ecological niche within the foreshore area of the beach where E. coli may survive and possibly multiply outside of host organisms. The results are of interest as increasing concentrations of E. coli in recreational waters are often considered to be an indication of recent fecal pollution.     

Pilot- and bench-scale testing of faecal indicator bacteria survival in marine beach sand near point sources

Aim:  Factors affecting faecal indicator bacteria (FIB) and pathogen survival/persistence in sand remain largely unstudied. This work elucidates how biological and physical factors affect die-off in beach sand following sewage spills.
Methods and Results:  Solar disinfection with mechanical mixing was pilot-tested as a disinfection procedure after a large sewage spill in Los Angeles. Effects of solar exposure, mechanical mixing, predation and/or competition, season, and moisture were tested at bench scale. First-order decay constants for Escherichia coli ranged between −0·23 and −1·02 per day, and for enterococci between −0·5 and −1·0 per day. Desiccation was a dominant factor for E. coli but not enterococci inactivation. Effects of season were investigated through a comparison of experimental results from winter, spring, and fall.
Conclusions:  Moisture was the dominant factor controlling E. coli inactivation kinetics. Initial microbial community and sand temperature were also important factors. Mechanical mixing, common in beach grooming, did not consistently reduce bacterial levels.
Significance and Impact of the Study:  Inactivation rates are mainly dependent on moisture and high sand temperature. Chlorination was an effective disinfection treatment in sand microcosms inoculated with raw influent.     

Abundance and characteristics of the recreational water quality indicator bacteria Escherichia coli and enterococci in gull faeces

AIMS: To evaluate the numbers and selected phenotypic and genotypic characteristics of the faecal indicator bacteria Escherichia coli and enterococci in gull faeces at representative Great Lakes swimming beaches in the United States. METHODS AND RESULTS: E. coli and enterococci were enumerated in gull faeces by membrane filtration. E. coli genotypes (rep-PCR genomic profiles) and E. coli (Vitek GNI+) and enterococci (API rapid ID 32 Strep and resistance to streptomycin, gentamicin, vancomycin, tetracycline and ampicillin) phenotypes were determined for isolates obtained from gull faeces both early and late in the swimming season. Identical E. coli genotypes were obtained only from single gull faecal samples but most faecal samples yielded more than one genotype (median of eight genotypes for samples with 10 isolates). E. coli isolates from the same site that clustered at >/=85% similarity were from the same sampling date and shared phenotypic characteristics, and at this similarity level there was population overlap between the two geographically isolated beach sites. Enterococcus API(R) profiles varied with sampling date. Gull enterococci displayed wide variation in antibiotic resistance patterns, and high-level resistance to some antibiotics. CONCLUSIONS: Gull faeces could be a major contributor of E. coli (10(5)-10(9) CFU g(-1)) and enterococci (10(4)-10(8) CFU g(-)1) to Great Lakes recreational waters. E. coli and enterococci in gull faeces are highly variable with respect to their genotypic and phenotypic characteristics and may exhibit temporal or geographic trends in these features. SIGNIFICANCE AND IMPACT OF THE STUDY: The high degree of variation in genotypic or phenotypic characteristics of E. coli or enterococci populations within gull hosts will require extensive sampling for adequate characterization, and will influence methods that use these characteristics to determine faecal contamination sources for recreational waters.     

A virulence and antimicrobial resistance DNA microarray detects a high frequency of virulence genes in Escherichia coli isolates from Great Lakes recreational waters

Escherichia coli is generally described as a commensal species with occasional pathogenic strains. Due to technological limitations, there is currently little information concerning the prevalence of pathogenic E. coli strains in the environment. For the first time, using a DNA microarray capable of detecting all currently described virulence genes and commonly found antimicrobial resistance genes, a survey of environmental E. coli isolates from recreational waters was carried out. A high proportion (29%) of 308 isolates from a beach site in the Great Lakes carried a pathotype set of virulence-related genes, and 14% carried antimicrobial resistance genes, findings consistent with a potential risk for public health. The results also showed that another 8% of the isolates had unusual virulence gene combinations that would be missed by conventional screening. This new application of a DNA microarray to environmental waters will likely have an important impact on public health, epidemiology, and microbial ecology in the future.     

Cladophora (Chlorophyta) spp. harbor human bacterial pathogens in nearshore water of Lake Michigan

Cladophora glomerata, a macrophytic green alga, is commonly found in the Great Lakes, and significant accumulations occur along shorelines during the summer months. Recently, Cladophora has been shown to harbor high densities of the fecal indicator bacteria Escherichia coli and enterococci. Cladophora may also harbor human pathogens; however, until now, no studies to address this question have been performed. In the present study, we determined whether attached Cladophora, obtained from the Lake Michigan and Burns Ditch (Little Calumet River, Indiana) sides of a breakwater during the summers of 2004 and 2005, harbored the bacterial pathogens Shiga toxin-producing Escherichia coli (STEC), Salmonella, Shigella, and Campylobacter. The presence of potential pathogens and numbers of organisms were determined by using cultural methods and by using conventional PCR, most-probable-number PCR (MPN-PCR), and quantitative PCR (QPCR) performed with genus- and toxin-specific primers and probes. While Shigella and STEC were detected in 100% and 25%, respectively, of the algal samples obtained near Burns Ditch in 2004, the same pathogens were not detected in samples collected in 2005. MPN-PCR and QPCR allowed enumeration of Salmonella in 40 to 80% of the ditch- and lakeside samples, respectively, and the densities were up to 1.6 x 10(3) cells per g Cladophora. Similarly, these PCR methods allowed enumeration of up to 5.4 x 10(2) Campylobacter cells/g Cladophora in 60 to 100% of lake- and ditchside samples. The Campylobacter densities were significantly higher (P < 0.05) in the lakeside Cladophora samples than in the ditchside Cladophora samples. DNA fingerprint analyses indicated that genotypically identical Salmonella isolates were associated with geographically and temporally distinct Cladophora samples. However, Campylobacter isolates were genetically diverse. Since animal hosts are thought to be the primary habitat for Campylobacter and Salmonella species, our results suggest that Cladophora is a likely secondary habitat for pathogenic bacteria in Lake Michigan and that the association of these bacteria with Cladophora warrants additional studies to assess the potential health impact on beach users.     

Clonal populations of thermotolerant Enterobacteriaceae in recreational water and their potential interference with fecal Escherichia coli counts

Bacterial strains were isolated from beach water samples using the original Environmental Protection Agency method for Escherichia coli enumeration and analyzed by pulsed-field gel electrophoresis (PFGE). Identical PFGE patterns were found for numerous isolates from 4 of the 9 days sampled, suggesting environmental replication. 16S rRNA gene sequencing, API 20E biochemical testing, and the absence of beta-glucuronidase activity revealed that these clonal isolates were Klebsiella, Citrobacter, and Enterobacter spp. In contrast, 82% of the nonclonal isolates from water samples were confirmed to be E. coli, and 16% were identified as other fecal coliforms. These nonclonal isolates produced a diverse range of PFGE patterns similar to those of isolates obtained directly from untreated sewage and gull droppings. beta-Glucuronidase activity was critical in distinguishing E. coli from other fecal coliforms, particularly for the clonal isolates. These findings demonstrate that E. coli is a better indicator of fecal pollution than fecal coliforms, which may replicate in the environment and falsely elevate indicator organism levels.     

Examination of the watershed-wide distribution of Escherichia coli along Southern Lake Michigan: an integrated approach

Recent research has highlighted the occurrence of Escherichia coli in natural habitats not directly influenced by sewage inputs. Most studies on E. coli in recreational water typically focus on discernible sources (e.g., effluent discharge and runoff) and fall short of integrating riparian, nearshore, onshore, and outfall sources. An integrated "beachshed" approach that links E. coli inputs and interactions would be helpful to understand the difference between background loading and sewage pollution; to develop more accurate predictive models; and to understand the differences between potential, net, and apparent culturable E. coli. The objective of this study was to examine the interrelatedness of E. coli occurrence from various coastal watershed components along southern Lake Michigan. The study shows that once established in forest soil, E. coli can persist throughout the year, potentially acting as a continuous non-point source of E. coli to nearby streams. Year-round background stream loading of E. coli can influence beach water quality. E. coli is present in highly variable counts in beach sand to depths just below the water table and to distances at least 5 m inland from the shore, providing a large potential area of input to beach water. In summary, E. coli in the fluvial-lacustrine system may be stored in forest soils, sediments surrounding springs, bank seeps, stream margins and pools, foreshore sand, and surface groundwater. While rainfall events may increase E. coli counts in the foreshore sand and lake water, concentrations quickly decline to prerain concentrations. Onshore winds cause an increase in E. coli in shallow nearshore water, likely resulting from resuspension of E. coli-laden beach sand. When examining indicator bacteria source, flux, and context, the entire "beachshed" as a dynamic interacting system should be considered.     

Foreshore Sand as a Source of Escherichia coli in Nearshore Water of a Lake Michigan Beach

Swimming advisories due to excessive Escherichia coli concentrations are common at 63rd Street Beach, Chicago, Ill. An intensive study was undertaken to characterize the source and fate of E. coli in beach water and sand at the beach. From April through September 2000, water and sand samples were collected daily or twice daily at two depths on three consecutive days per week (water samples, n = 1,747; sand samples, n = 858); hydrometeorological conditions and bird and bather distributions were also recorded. E. coli concentrations in sand and water were significantly correlated, with the highest concentration being found in foreshore sand, followed by those in submerged sediment and water of increasing depth. Gull contributions to E. coli densities in sand and water were most apparent on the day following gull activity in a given area. E. coli recolonized newly placed foreshore sand within 2 weeks. Analysis of variance, correlation, cluster analyses, concentration gradients, temporal-spatial distribution, demographic patterns, and DNA fingerprinting suggest that E. coli may be able to sustain population density in temperate beach sand during summer months without external inputs. This research presents evidence that foreshore beach sand (i) plays a major role in bacterial lake water quality, (ii) is an important non-point source of E. coli to lake water rather than a net sink, (iii) may be environmentally, and perhaps hygienically, problematic, and (iv) is possibly capable of supporting an autochthonous, high density of indicator bacteria for sustained periods, independent of lake, human, or animal input.     

Seed Mass Effects Across Environments in an Annual Dune Plant

The natural environment has been shown to vary at both large and small scales. As the relationship between seed mass and the growth of subsequent plants varies depending on the environment, it is important to study the effects of seed mass on plant growth across environments for a better understanding of the ecological significance of seed mass. In this study, the effects of seed mass on the growth ofCakile edentula, an annual growing in the sand dunes along the Great Lakes, was examined across a wide range of soil nutrient, moisture levels, and depths of sand burial under controlled greenhouse conditions. The development of plants, such as time of true leaf production, flower bud formation, and fruit maturation, was faster in proportion to the original seed mass of the plants across all of the tested environmental factors. As drastic environmental variations prevail in beach sand dune systems, quick development and early reproductive growth of plants from the large seeds would enable them to survive sand burial and escape natural hazards. It is concluded that seed mass has significant effects on the performance ofC. edentulaacross a wide range of environmental variation.     

Sand and nest temperatures and an estimate of hatchling sex ratio from the Heron Island green turtle (Chelonia mydas) rookery, Southern Great Barrier Reef

Sand and nest temperatures were monitored during the 2002–2003 nesting season of the green turtle, Chelonia mydas, at Heron Island, Great Barrier Reef, Australia. Sand temperatures increased from ∼ 24°C early in the season to 27–29°C in the middle, before decreasing again. Beach orientation affected sand temperature at nest depth throughout the season; the north facing beach remained 0.7°C warmer than the east, which was 0.9°C warmer than the south, but monitored nest temperatures were similar across all beaches. Sand temperature at 100 cm depth was cooler than at 40 cm early in the season, but this reversed at the end. Nest temperatures increased 2–4°C above sand temperatures during the later half of incubation due to metabolic heating. Hatchling sex ratio inferred from nest temperature profiles indicated a strong female bias.     

New method for estimating bacterial cell abundances in natural samples by use of sublimation

We have developed a new method based on the sublimation of adenine from Escherichia coli to estimate bacterial cell counts in natural samples. To demonstrate this technique, several types of natural samples, including beach sand, seawater, deep-sea sediment, and two soil samples from the Atacama Desert, were heated to a temperature of 500 degrees C for several seconds under reduced pressure. The sublimate was collected on a cold finger, and the amount of adenine released from the samples was then determined by high-performance liquid chromatography with UV absorbance detection. Based on the total amount of adenine recovered from DNA and RNA in these samples, we estimated bacterial cell counts ranging from approximately 10(5) to 10(9) E. coli cell equivalents per gram. For most of these samples, the sublimation-based cell counts were in agreement with total bacterial counts obtained by traditional DAPI (4,6-diamidino-2-phenylindole) staining.     

Ubiquity and persistence of Escherichia coli in a Midwestern coastal stream

Dunes Creek, a small Lake Michigan coastal stream that drains sandy aquifers and wetlands of Indiana Dunes, has chronically elevated Escherichia coli levels along the bathing beach near its outfall. This study sought to understand the sources of E. coli in Dunes Creek's central branch. A systematic survey of random and fixed sampling points of water and sediment was conducted over 3 years. E. coli concentrations in Dunes Creek and beach water were significantly correlated. Weekly monitoring at 14 stations during 1999 and 2000 indicated chronic loading of E. coli throughout the stream. Significant correlations between E. coli numbers in stream water and stream sediment, submerged sediment and margin, and margin and 1 m from shore were found. Median E. coli counts were highest in stream sediments, followed by bank sediments, sediments along spring margins, stream water, and isolated pools; in forest soils, E. coli counts were more variable and relatively lower. Sediment moisture was significantly correlated with E. coli counts. Direct fecal input inadequately explains the widespread and consistent occurrence of E. coli in the Dunes Creek watershed; long-term survival or multiplication or both seem likely. The authors conclude that (i) E. coli is ubiquitous and persistent throughout the Dunes Creek basin, (ii) E. coli occurrence and distribution in riparian sediments help account for the continuous loading of the bacteria in Dunes Creek, and (iii) ditching of the stream, increased drainage, and subsequent loss of wetlands may account for the chronically high E. coli levels observed.     

Detection of Genetic Markers of Fecal Indicator Bacteria in Lake Michigan and Determination of Their Relationship to Escherichia coli Densities Using Standard Microbiological Methods

Lake Michigan surface waters impacted by fecal pollution were assessed to determine the occurrence of genetic markers for Bacteroides and Escherichia coli. Initial experiments with sewage treatment plant influent demonstrated that total Bacteroides spp. could be detected by PCR in a 25- to 125-fold-higher dilution series than E. coli and human-specific Bacteroides spp., which were both found in similar dilution ranges. The limit of detection for the human-specific genetic marker ranged from 0.2 CFU/100 ml to 82 CFU/100 ml culturable E. coli for four wastewater treatment plants in urban and rural areas. The spatial and temporal distributions of these markers were assessed following major rain events that introduced urban storm water, agricultural runoff, and sewage overflows into Lake Michigan. Bacteroides spp. were detected in all of these samples by PCR, including those with <1 CFU/100 ml E. coli. Human-specific Bacteroides spp. were detected as far as 2 km into Lake Michigan during sewage overflow events, with variable detection 1 to 9 days postoverflow, whereas the cow-specific Bacteroides spp. were detected in only highly contaminated samples near the river outflow. Lake Michigan beaches were also assessed throughout the summer season for the same markers. Bacteroides spp. were detected in all beach samples, including 28 of the 74 samples that did not exceed 235 CFU/100 ml of E. coli. Human-specific Bacteroides spp. were detected at three of the seven beaches; one of the sites demonstrating positive results was sampled during a reported sewage overflow, but E. coli levels were below 235 CFU/100 ml. This study demonstrates the usefulness of non-culture-based microbial-source tracking approaches and the prevalence of these genetic markers in the Great Lakes, including freshwater coastal beaches.     

Genetic Relatedness of Escherichia coli Isolates in Interstitial Water from a Lake Huron (Canada) Beach

Research was undertaken to characterize Escherichia coli isolates in interstitial water samples of a sandy beach on the southeastern shore of Lake Huron, Ontario, Canada. A survey of the beach area revealed the highest abundance of E. coli in interstitial water of the foreshore beach sand next to the swash zone. Higher concentrations of E. coli (up to 1.6 × 10⁶ CFU/100 ml of water) were observed in the interstitial water from the sampling holes on the beach itself compared to lake water and sediment. Repetitive extragenic palindromic PCR (REP-PCR) was used to characterize the genetic diversity of E. coli isolates from interstitial water samples on the beach. E. coli isolates from the same sampling location frequently exhibited the same REP-PCR pattern or were highly similar to each other. In contrast, E. coli isolates from different sampling locations represented populations distinct from each other. This study has identified a unique ecological niche within the foreshore area of the beach where E. coli may survive and possibly multiply outside of host organisms. The results are of interest as increasing concentrations of E. coli in recreational waters are often considered to be an indication of recent fecal pollution.     

Soil survival of Escherichia coli O157:H7 acquired by a child from garden soil recently fertilized with cattle manure

AIMS: This investigation was conducted to determine the survival of a naturally occurring Escherichia coli O157:H7 in garden soil linked to a sporadic case of E. coli O157 infection in Minnesota. METHODS AND RESULTS: The presence and viability of E. coli O157:H7 was monitored in manure-contaminated garden soil for several weeks. Bacterial isolates were characterized using PCR and pulsed-field gel electrophoresis (PFGE). Isolates obtained from the patient and the garden plots during this investigation had indistinguishable PFGE patterns and had the same virulence factors (stx1, stx2, eaeA, ehxA). The E. coli O157:H7 levels obtained from the garden plots declined gradually for a period of 2 months, and on day 69 only one garden plot of four had detectable levels of pathogen. All plots were negative on day 92. The rate of decline in the soil samples stored at 4 degrees C was faster compared with soil samples that remained in ambient conditions, and in refrigerated storage E. coli O157:H7 could not be detected after 10 days. CONCLUSIONS: E. coli O157:H7 strains can survive on manure-amended soil for more than 2 months, and this survival could be reduced by low temperature. SIGNIFICANCE AND IMPACT OF THE STUDY: This is one of the few reports that have investigated the survival of a proven virulent strain in naturally contaminated soil samples. This case stresses the importance of avoiding the use of raw cattle manure to amend soil for cultivation of foods, including soils in residential garden plots.     

Escherichia coli contamination of vegetables grown in soils fertilized with noncomposted bovine manure: garden-scale studies

In this study we tested the validity of the National Organic Program (NOP) requirement for a > or =120-day interval between application of noncomposted manure and harvesting of vegetables grown in manure-fertilized soil. Noncomposted bovine manure was applied to 9.3-m2 plots at three Wisconsin sites (loamy sand, silt loam, and silty clay loam) prior to spring and summer planting of carrots, radishes, and lettuce. Soil and washed (30 s under running tap water) vegetables were analyzed for indigenous Escherichia coli. Within 90 days, the level of E. coli in manure-fertilized soil generally decreased by about 3 log CFU/g from initial levels of 4.2 to 4.4 log CFU/g. Low levels of E. coli generally persisted in manure-fertilized soil for more than 100 days and were detected in enriched soil from all three sites 132 to 168 days after manure application. For carrots and lettuce, at least one enrichment-negative sample was obtained < or =100 days after manure application for 63 and 88% of the treatments, respectively. The current > or =120-day limit provided an even greater likelihood of not detecting E. coli on carrots (> or =1 enrichment-negative result for 100% of the treatments). The rapid maturation of radishes prevented conclusive evaluation of a 100- or 120-day application-to-harvest interval. The absolute absence of E. coli from vegetables harvested from manure-fertilized Wisconsin soils may not be ensured solely by adherence to the NOP > or =120-day limit. Unless pathogens are far better at colonizing vegetables than indigenous E. coli strains are, it appears that the risk of contamination for vegetables grown in Wisconsin soils would be elevated only slightly by reducing the NOP requirement to > or =100 days.     

General suppression of Escherichia coli O157:H7 in sand-based dairy livestock bedding

Sand bedding material is frequently used in dairy operations to reduce the occurrence of mastitis and enhance cow comfort. One objective of this work was to determine if sand-based bedding also supported the microbiologically based suppression of an introduced bacterial pathogen. Bedding samples were collected in summer, fall, and winter from various locations within a dairy operation and tested for their ability to suppress introduced populations of Escherichia coli O157:H7. All sources of bedding displayed a heat-sensitive suppressiveness to the pathogen. Differences in suppressiveness were also noted between different samples at room temperature. At just 1 day postinoculation (dpi), the recycled sand bedding catalyzed up to a 1,000-fold reduction in E. coli counts, typically 10-fold greater than the reduction achieved with other substrates, depending on the sampling date. All bedding substrates were able to reduce E. coli populations by over 10,000-fold within 7 to 15 dpi, regardless of sampling date. Terminal restriction fragment length polymorphism (T-RFLP) analysis was used to identify bacterial populations potentially associated with the noted suppression of E. coli O157:H7 in sand bedding. Eleven terminal restriction fragments (TRFs) were overrepresented in paired comparisons of suppressive and nonsuppressive specimens at multiple sampling points, indicating that they may represent environmentally stable populations of pathogen-suppressing bacteria. Cloning and sequencing of these TRFs indicated that they represent a diverse subset of bacteria, belonging to the Cytophaga-Flexibacter-Bacteroidetes, Gammaproteobacteria, and Firmicutes, only a few of which have previously been identified in livestock manure. Such data indicate that microbial suppression may be harnessed to develop new options for mitigating the risk and dispersal of zoonotic bacterial pathogens on dairy farms.     

Occurrence and persistence of bacterial pathogens and indicator organisms in beach sand along the California coast

This report documents the presence of fecal indicators and bacterial pathogens in sand at 53 California marine beaches using both culture-dependent and -independent (PCR and quantitative PCR [QPCR]) methods. Fecal indicator bacteria were widespread in California beach sand, with Escherichia coli and enterococci detected at 68% and 94% of the beaches surveyed, respectively. Somatic coliphages and a Bacteroidales human-specific fecal marker were detected at 43% and 13% of the beaches, respectively. Dry sand samples from almost 30% of the beaches contained at least one of the following pathogens: Salmonella spp., Campylobacter spp., Staphylococcus aureus, and methicillin-resistant Staphylococcus aureus (MRSA), which were detected at 15%, 13%, 14%, and 3% of tested beaches, respectively. Fecal indicators and pathogens were poorly correlated to one another and to land cover. Sands were dry at the time of collection, and those with relatively high moisture tended to have higher concentrations or a more frequent occurrence of both indicators and pathogens. Using culture-dependent assays, fecal indicators decayed faster than pathogens in microcosm experiments using unaltered beach sand seeded with sewage and assessed by culture-dependent assays. The following order of persistence was observed (listed from most to least persistent): Campylobacter > Salmonella > somatic coliphages > enterococci > E. coli > F(+) phages. In contrast, pathogens decayed faster than fecal indicators in culture-independent assays: enterococci > Bacteroidales human-specific marker > Salmonella > Campylobacter. Microcosm experiments demonstrated that both indicators and pathogens were mobilized by wetting with seawater. Decay rates measured by QPCR were lower than those measured with culture-dependent methods. Enterococcal persistence and possible growth were observed for wetted microcosms relative to unwetted controls.