Geology topics

Muruleedhara N. Byappanahalli

Publications and source records attributed to Muruleedhara N. Byappanahalli.

31 records · Page 2Linked to original sources

USGS2013

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

Here we report results from a multi-laboratory ( n = 11) evaluation of four different PCR methods targeting the 16S rRNA gene of Catellicoccus marimammalium originally developed to detect gull fecal contamination in coastal environments. The methods included a conventional end-point PCR method, a SYBR ® Green qPCR method, and two TaqMan ® qPCR methods. Different techniques for data normalization and analysis were tested. Data analysis methods had a pronounced impact on assay sensitivity and specificity calculations. Across-laboratory standardization of metrics including the lower limit of quantification (LLOQ), target detected but not quantifiable (DNQ), and target not detected (ND) significantly improved results compared to results submitted by individual laboratories prior to definition standardization. The unit of measure used for data normalization also had a pronounced effect on measured assay performance. Data normalization to DNA mass improved quantitative method performance as compared to enterococcus normalization. The MST methods tested here were originally designed for gulls but were found in this study to also detect feces from other birds, particularly feces composited from pigeons. Sequencing efforts showed that some pigeon feces from California contained sequences similar to C. marimammalium found in gull feces. These data suggest that the prevalence, geographic scope, and ecology of C. marimammalium in host birds other than gulls require further investigation. This study represents an important first step in the multi-laboratory assessment of these methods and highlights the need to broaden and standardize additional evaluations, including environmentally relevant target concentrations in ambient waters from diverse geographic regions.

Water Research↗

USGS2013

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

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.

Water Research↗

USGS2012

Nearshore hydrodynamics as loading and forcing factors for Escherichia coli contamination at an embayed beach

Numerical simulations of the transport and fate of Escherichia coli were conducted at Chicago's 63rd Street Beach, an embayed beach that had the highest mean E. coli concentration among 23 similar Lake Michigan beaches during summer months of 2000-2005, in order to find the cause for the high bacterial contamination. The numerical model was based on the transport of E. coli by current circulation patterns in the embayment driven by longshore main currents and the loss of E. coli in the water column, taking settling as well as bacterial dark- and solar-related decay into account. Two E. coli loading scenarios were considered: one from the open boundary north of the embayment and the other from the shallow water near the beachfront. Simulations showed that the embayed beach behaves as a sink for E. coli in that it generally receives E. coli more efficiently than it releases them. This is a result of the significantly different hydrodynamic forcing factors between the inside of the embayment and the main coastal flow outside. The settled E. coli inside the embayment can be a potential source of contamination during subsequent sediment resuspension events, suggesting that deposition-resuspension cycles of E. coli have resulted in excessive bacterial contamination of beach water. A further hypothetical case with a breakwater shortened to half its original length, which was anticipated to enhance the current circulation in the embayment, showed a reduction in E. coli concentrations of nearly 20%.

Illinois↗

USGS2012

Evidence for occurrence, persistence, and growth potential of Escherichia coli and enterococci in Hawaii’s soil environments

High densities of Escherichia coli and enterococci are common in freshwaters on Oahu and other Hawaiian Islands. Soil along stream banks has long been suspected as the likely source of these bacteria; however, the extent of their occurrence and distribution in a wide range of soils remained unknown until the current investigation. Soil samples representing the seven major soil associations were collected on the island of Oahu and analyzed for fecal coliforms, E. coli , and enterococci by the most probable number method. Fecal coliforms, E. coli , and enterococci were found in most of the samples analyzed; log mean densities (MPN ± SE g soil −1 ) were 1.96±0.18, n =61; 1.21±0.17, n =57; and 2.99±0.12, n =62, respectively. Representative, presumptive cultures of E. coli and enterococci collected from the various soils were identified and further speciated using the API scheme; at least six species of Enterococcus , including Enterococcus faecalis and Enterococcus faecium , were identified. In mesocosm studies, E. coli and enterococci increased by 100-fold in 4 days, after mixing sewage-spiked soil (one part) with autoclaved soil (nine parts). E. coli remained metabolically active in the soil and readily responded to nutrients, as evidenced by increased dehydrogenase activity. Collectively, these findings indicate that populations of E. coli and enterococci are part of the natural soil microflora, potentially influencing the quality of nearby water bodies.

Hawai'i↗

USGS2011

Environmental sources of fecal bacteria

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

Book chapter↗

USGS2011

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

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

Book chapter↗

USGS2010

Relationship and variation of qPCR and culturable enterococci estimates in ambient surface waters are predictable

The quantitative polymerase chain reaction (qPCR) method provides rapid estimates of fecal indicator bacteria densities that have been indicated to be useful in the assessment of water quality. Primarily because this method provides faster results than standard culture-based methods, the U.S. Environmental Protection Agency is currently considering its use as a basis for revised ambient water quality criteria. In anticipation of this possibility, we sought to examine the relationship between qPCR-based and culture-based estimates of enterococci in surface waters. Using data from several research groups, we compared enterococci estimates by the two methods in water samples collected from 37 sites across the United States. A consistent linear pattern in the relationship between cell equivalents (CCE), based on the qPCR method, and colony-forming units (CFU), based on the traditional culturable method, was significant (P < 0.05) at most sites. A linearly decreasing variance of CCE with increasing CFU levels was significant (P < 0.05) or evident for all sites. Both marine and freshwater sites under continuous influence of point-source contamination tended to reveal a relatively constant proportion of CCE to CFU. The consistency in the mean and variance patterns of CCE versus CFU indicates that the relationship of results based on these two methods is more predictable at high CFU levels (e.g., log 10 CFU > 2.0/100 mL) while uncertainty increases at lower CFU values. It was further noted that the relative error in replicated qPCR estimates was generally higher than that in replicated culture counts even at relatively high target levels, suggesting a greater need for replicated analyses in the qPCR method to reduce relative error. Further studies evaluating the relationship between culture and qPCR should take into account analytical uncertainty as well as potential differences in results of these methods that may arise from sample variability, different sources of pollution, and environmental factors.

Environmental Science & Technology↗

USGS2010

Linking non-culturable (qPCR) and culturable enterococci densities with hydrometeorological conditions

Quantitative polymerase chain reaction (qPCR) measurement of enterococci has been proposed as a rapid technique for assessment of beach water quality, but the response of qPCR results to environmental conditions has not been fully explored. Culture-based E. coli and enterococci have been used in empirical predictive models to characterize their responses to environmental conditions and to increase monitoring frequency and efficiency. This approach has been attempted with qPCR results only in few studies. During the summer of 2006, water samples were collected from two southern Lake Michigan beaches and the nearby river outfall (Burns Ditch) and were analyzed for enterococci by culture-based and non-culture-based (i.e., qPCR) methods, as well as culture-based E. coli . Culturable enterococci densities (log CFU/100 ml) for the beaches were significantly correlated with enterococci qPCR cell equivalents (CE) ( R = 0.650, P < 0.0001, N = 32). Enterococci CE and CFU densities were highest in Burns Ditch relative to the beach sites; however, only CFUs were significantly higher ( P < 0.0001). Culturable enterococci densities at Burns Ditch and the beaches were significantly correlated ( R = 0.565, P < 0.0001, N = 32). Culturable E. coli and enterococci densities were significantly correlated ( R = 0.682, P < 0.0001, N = 32). Regression analyses suggested that enterococci CFU could be predicted by lake turbidity, Burns Ditch discharge, and wind direction (adjusted R 2 = 0.608); enterococci CE was best predicted by Burns Ditch discharge and log-transformed lake turbidity × wave height (adjusted R 2 = 0.40). In summary, our results show that analytically, the qPCR method compares well to the non-culture-based method for measuring enterococci densities in beach water and that both these approaches can be predicted by hydrometeorological conditions. Selected predictors and model results highlight the differences between the environmental responses of the two method endpoints and the potentially high variance in qPCR results

Lake Michigan↗

USGS2010

Cladophora in the Great Lakes: Impacts on beach water quality and human health

Cladophora in the Great Lakes grows rapidly during the warm summer months, detaches, and becomes free-floating mats as a result of environmental conditions, eventually becoming stranded on recreational beaches. Cladophora provides protection and nutrients, which allow enteric bacteria such as Escherichia coli , enterococci, Shigella , Campylobacter , and Salmonella to persist and potentially regrow in the presence of the algae. As a result of wind and wave action, these microorganisms can detach and be released to surrounding waters and can influence water quality. Enteric bacterial pathogens have been detected in Cladophora mats; E. coli and enterococci may populate to become part of the naturalized microbiota in Cladophora ; the high densities of these bacteria may affect water quality, resulting in unnecessary beach closures. The continued use of traditional fecal indicators at beaches with Cladophora presence is inadequate at accurately predicting the presence of fecal contamination. This paper offers a substantial review of available literature to improve the knowledge of Cladophora impacts on water quality, recreational water monitoring, fecal indicator bacteria and microorganisms, and public health and policy.

Water Science and Technology↗

USGS2004

Solar and temporal effects on Escherichia coli concentration at a Lake Michigan swimming beach

Studies on solar inactivation of Escherichia coli in freshwater and in situ have been limited. At 63rd St. Beach, Chicago, Ill., factors influencing the daily periodicity of culturable E. coli , particularly insolation, were examined. Water samples for E. coli analysis were collected twice daily between April and September 2000 three times a week along five transects in two depths of water. Hydrometeorological conditions were continuously logged: UV radiation, total insolation, wind speed and direction, wave height, and relative lake level. On 10 days, transects were sampled hourly from 0700 to 1500 h. The effect of sunlight on E. coli inactivation was evaluated with dark and transparent in situ mesocosms and ambient lake water. For the study, the number of E. coli samples collected ( n ) was 2,676. During sunny days, E. coli counts decreased exponentially with day length and exposure to insolation, but on cloudy days, E. coli inactivation was diminished; the E. coli decay rate was strongly influenced by initial concentration. In situ experiments confirmed that insolation primarily inactivated E. coli ; UV radiation only marginally affected E. coli concentration. The relationship between insolation and E. coli density is complicated by relative lake level, wave height, and turbidity, all of which are often products of wind vector. Continuous importation and nighttime replenishment of E. coli were evident. These findings (i) suggest that solar inactivation is an important mechanism for natural reduction of indicator bacteria in large freshwater bodies and (ii) have implications for management strategies of nontidal waters and the use of E. coli as an indicator organism.

Applied and Environmental Microbiology↗

USGS2003

Evaluation of beach grooming techniques on Escherichia coli density in foreshore sand at North Beach, Racine, WI

Elevated levels of Escherichia coli(E. coli) in bathing waters at North Beach, a popular recreational site in Racine, Wisconsin, have been a persistent problem often resulting in the issuance of poor water quality advisories. Moreover, waterfowl (mostly Larus delawarensis and L. argentatus ) in nearshore and offshore areas are common and may serve as non-point sources for bacterial contamination of recreational waters. Current beach management practice involves daily mechanical grooming of the nearshore sand for aesthetics and removal of hazardous debris. However, this practice has not been evaluated in terms of its effects on E. coli loading to beach sand and potential introduction to contiguous swimming water. In this study, we tested E. coli responses to three treatments: mechanical groomer, daily and twice weekly hand raking, and a control (no raking/grooming). A randomized block design consisted of replicated treatments and one control (10 each), for a total of 40 blocks sampled daily for 10 days. Foreshore sand samples were collected by hand coring to an average depth of 10 cm. Median E. coli recovered were 73 (mechanically groomed), 27 (hand-raked daily), 32 (hand-raked twice weekly), and 22 (control) colony-forming units (CFU) per gram dry weight sand. E. coli counts in sand that was groomed were significantly higher than hand rakings and control (p <0.0001), and there was no significant difference between control and raking treatments (p<0.01). This study demonstrates the beach management implications related to grooming efficacy and the importance of understanding non-point sources of bacterial contamination.

Lake and Reservoir Management↗

USGS2003

Evaluation of a rapid, quantitative real-time PCR method for enumeration of pathogenic Candida cells in water

Quantitative PCR (QPCR) technology, incorporating fluorigenic 5′ nuclease (TaqMan) chemistry, was utilized for the specific detection and quantification of six pathogenic species of Candida ( C. albicans , C. tropicalis , C. krusei , C. parapsilosis , C. glabrata and C. lusitaniae ) in water. Known numbers of target cells were added to distilled and tap water samples, filtered, and disrupted directly on the membranes for recovery of DNA for QPCR analysis. The assay's sensitivities were between one and three cells per filter. The accuracy of the cell estimates was between 50 and 200% of their true value (95% confidence level). In similar tests with surface water samples, the presence of PCR inhibitory compounds necessitated further purification and/or dilution of the DNA extracts, with resultant reductions in sensitivity but generally not in quantitative accuracy. Analyses of a series of freshwater samples collected from a recreational beach showed positive correlations between the QPCR results and colony counts of the corresponding target species. Positive correlations were also seen between the cell quantities of the target Candida species detected in these analyses and colony counts of Enterococcus organisms. With a combined sample processing and analysis time of less than 4 h, this method shows great promise as a tool for rapidly assessing potential exposures to waterborne pathogenic Candida species from drinking and recreational waters and may have applications in the detection of fecal pollution.

Applied and Environmental Microbiology↗

USGS2003

Occurrence of Escherichia coli and enterococci in Cladophora (Chlorophyta) in nearshore water and beach sand of Lake Michigan

Each summer, the nuisance green alga Cladophora (mostly Cladophora glomerata ) amasses along Lake Michigan beaches, creating nearshore anoxia and unsightly, malodorous mats that can attract problem animals and detract from visitor enjoyment. Traditionally, elevated counts of Escherichia coli are presumed to indicate the presence of sewage, mostly derived from nearby point sources. The relationship between fecal indicator bacteria and Cladophora remains essentially unstudied. This investigation describes the local and regional density of Escherichia coli and enterococci in Cladophora mats along beaches in the four states (Wisconsin, Illinois, Indiana, and Michigan) bordering Lake Michigan. Samples of Cladophora strands collected from 10 beaches ( n = 41) were assayed for concentrations of E. coli and enterococci during the summer of 2002. Both E. coli and enterococci were ubiquitous (up to 97% occurrence), with overall log mean densities (± standard errors) of 5.3 (± 4.8) and 4.8 (± 4.5) per g (dry weight). E. coli and enterococci were strongly correlated in southern Lake Michigan beaches ( P < 0.001, R 2 = 0.73, n = 17) but not in northern beaches ( P = 0.892, n = 16). Both E. coli and enterococci survived for over 6 months in sun-dried Cladophora mats stored at 4°C; the residual bacteria in the dried alga readily grew upon rehydration. These findings suggest that Cladophora amassing along the beaches of Lake Michigan may be an important environmental source of indicator bacteria and call into question the reliability of E. coli and enterococci as indicators of water quality for freshwater recreational beaches.

Applied and Environmental Microbiology↗