Risk-based evaluation of Escherichia coli monitoring data from undisinfected drinking water

Drinking water regulations in the United States and elsewhere are based on the occurrence of fecal indicator bacteria. Though not meeting all the criteria of an ideal indicator, nonpathogenic strains of Escherichia coli (E. coli) are used worldwide as an indicator of potential fecal contamination for drinking water and for distribution systems. This is, in part, because real illnesses are related to human pathogens, such as E. coli O157:H7, whose presence may be predicted better by E. coli than by total coliform bacteria. Our objective was to estimate the number of E. coli O157:H7 illnesses attributable to drinking water exposures in the United States and the feasible relationships between positive occurrences of the indicator bacteria E. coli and E. coli O157:H7 in drinking water. Results of the modeling indicate that in undisinfected drinking water systems, the ratio of bacterial indicator E. coli positives to E. coli O157:H7 organisms is estimated to be between 6:1 and 90:1 with few model parameters accounting for the vast majority of the uncertainty. These results provide context for considering the potential public health implications of a positive E. coli result from routine monitoring of undisinfected drinking water.     

ESCHERICHIA COLI SURVIVAL IN MANTLED KARST SPRINGS AND STREAMS,NORTHWEST ARKANSAS OZARKS,USA1

Recent studies indicate fecal coliform bacterial concentrations, including Escherichia coli (E. coli), characteristically vary by several orders of magnitude, depending on the hydrology of storm recharge and discharge. E. coli concentrations in spring water increase rapidly during the rising limb of a storm hydrograph, peak prior to or coincident with the peak of the storm pulse, and decline rapidly, well before the recession of the storm hydrograph. This suggests E. coli are associated with resuspension of sediment during the onset of turbulent flow, and indicates viable bacteria reside within the spring and stream sediments. E. coli inoculated chambers were placed in spring and stream environments within the mantled karst of northwest Arkansas to assess long term (> 75 days) E. coli viability. During the 75‐day study, a 4‐log die‐off of E. coli was observed for chambers placed in the Illinois River, and a 5‐log die‐off for chambers placed in Copperhead Spring. Extrapolation of the regression line for each environment indicates E. coli concentration would reach 1 most probable number (MPN)/100 g sediment at Copperhead Spring in about 105 days, and about 135 days in the Illinois River, based on a starting inoculation of 2.5 × 107 MPN E. coli/100 g of sediment. These in situ observations indicate it is possible for E. coli to survive in these environments for at least four months with no fresh external inputs.     

Fate and Transport Modeling of Potential Pathogens: The Contribution From Sediments1

Abstract: Escherichia coli was used as a bacterial tracer for the development, calibration, and validation of a watershed scale fate and transport model to be extended to a suite of reference pathogens (Cryptosporidium, Giardia, Campylobacter, E. coli O157:H7). E. coli densities in water and sediments from the Blackstone River Watershed, Massachusetts, were measured at three sites for a total of five wet weather events and three dry weather events covering three seasons. The confirmed E. coli strains were identified by ribotyping for tracking the sources of E. coli and for determining the association of downstream E. coli isolates with isolates from upstream sediments. A large number of downstream samples were associated with upstream sediment sources of E. coli. E. coli densities ranged from 71 to 6,401 MPN/100 ml in water samples and from 2 to 335 MPN/g in sediments. Pearson correlation analysis revealed significant correlations between E. coli and total coliforms in water (r = 0.777, p < 0.01) and sediments (r = 0.728, p < 0.01). In addition, E. coli concentrations in water were weakly correlated with sediment particle size and sediment concentrations (r = 0.298, p < 0.01). A hydrologic model, WATFLOOD/SPL9, was used to predict the temporal and spatial variation of E. coli in the Blackstone River. The rapid rise of stream E. coli densities was more accurately predicted by the model with the inclusion of sediment resuspension, thus demonstrating the importance of the process.     

Assessing the Impacts of E. coli Laden Streambed Sediment on E. coli Loads over a Range of Flows and Sediment Characteristics

Understanding sediment Escherichia coli levels (i.e., pathogen indicators) and their contribution to the water column during resuspension is critical for predicting in‐stream E. coli levels and the potential risk to human health. The U.S. Environmental Protection Agency's current water quality testing strategies, however, rely on water borne E. coli concentrations to assess stream E. coli levels and identify impaired waters. In this work, we conducted a scenario analysis using a range of flows, sediment/water bacteria fractions, and particle sizes to which E. coli attach to assess the impact of E. coli in streambed sediments on water column E. coli levels. We used simple sediment transport theory to calculate the potential total E. coli concentrations in a stream with and without the resuspension process. Results clearly indicate that inclusion of resuspending sediment attached E. coli is necessary for watershed assessments and data on sediment attached E. coli concentrations is much needed. When neglecting the streambed sediment E. coli concentrations, the model predicted average E. coli loads of 10⁷ Colony Forming Units (CFU)/s; however, when streambed sediment E. coli concentrations were included in the model, the predictions ranged from 10¹⁰ to 10¹⁴ CFU/s. To evaluate the predictions, E. coli data in the streambed sediment and the water column were monitored in Squaw Creek, Iowa. Comparisons between measured and predicted E. coli loads yielded an R²‐value of 0.85.     

ACCOMMODATING CHANGE OF BACTERIAL INDICATORS IN LONG TERM WATER QUALITY DATASETS1

ABSTRACT: In 1996, the State of Oregon adopted a water quality standard based on Escherichia coli (E. coli), recognizing E. coli as an indicator of pathogenic potential. The Oregon Department of Environmental Quality (DEQ) began analysis for E. coli that same year. The Oregon DEQ continued collection and analysis of fecal coliform (a prior indicator organism) for data input to bacterial loading models and the Oregon Water Quality Index (OWQI). The OWQI is a primary indicator of general water quality for the Oregon DEQ and the Oregon Progress Board. The objective of this study was to develop a regression relationship between fecal coliform and E. coli. This relationship would fill data gaps and extend water quality models and indicators. Water quality policy is better informed by the ability of these extended water quality models to determine whether water quality meets present or would have met past bacterial standards. Monitoring resources spent on dual bacterial analyses could be conserved. This study also showed that changes to OWQI values (as a result of changing bacterial indicators) were minimal, and corresponded to improved characterization of water quality with respect to pathogenic potential.     

Escherichia coli Concentrations in Urban Watersheds Exhibit Diurnal Sag: Implications for Water‐Quality Monitoring and Assessment

The variability of indicator bacteria over a fine resolution time scale on the order of minutes has yet to be fully understood. In this study, we collected more than 700 Escherichia coli samples at a 10‐ and 30‐min resolution in an urban watershed in Houston. A Bacteria Diurnal Sag (BDS) marked with daytime exponential decay followed by an exponential nighttime regeneration was observed. This pattern was observed during all sampled events but varied depending on other variables. The concentrations during a 24‐h period varied 1 to 5 orders of magnitude and the fecal load was at least 10 times lower than what would be obtained using a single morning E. coli measurement, the typical sampling scheme in most monitoring programs. Decay rates, ranging from 3.67 to 24.7/day, decreased E. coli concentrations to below the water‐quality standards from 14:00 to 18:00 h and were strongly influenced by water temperatures and solar radiation intensities. Rapid regeneration occurred on the order of 9.41 to 64.1/day allowing E. coli concentrations to return to their pre‐decay levels. The data indicated that four to six samples taken between 06:00 and 18:00 h may be sufficient to define the BDS depending on stream conditions, and that a threshold concentration of approximately 100 MPN/dl (most probable number in a deciliter) existed for the studied urban watershed. These findings have significant implications for water‐quality monitoring, regulation, and compliance.     

Cattle feedlot soil moisture and manure content: II. Impact on Escherichia coli O157

The moisture and manure contents of soils at cattle feedlot surfaces vary spatiotemporally and likely are important factors in the persistence of Escherichia coli O157 in these soils. The impacts of water content (0.11-1.50 g H2O g(-1) dry feedlot surface material [FSM]) and manure level (5, 25, and 75% dry manure in dry FSM) on E. coli O157:H7 in feedlot soils were evaluated. Generally, E. coli O157:H7 numbers either persisted or increased at all but the lowest moisture levels examined. Manure content modulated the effect of water on E. coli growth; for example, at water content of 0.43 g H2O g(-1) dry FSM and 25% manure, E. coli O157:H7 increased by 2 log10 colony forming units (CFU) g(-1) dry FSM in 3 d, while at 0.43 g H2O g(-1) dry FSM and 75% manure, populations remained stable over 14 d. Escherichia coli and coliform populations responded similarly. In a second study, the impacts of cycling moisture levels and different drying rates on naturally occurring E. coli O157 in feedlot soils were examined. Low initial levels of E. coli O157 were reduced to below enumerable levels by 21 d, but indigenous E. coli populations persisted at >2.50 log10 CFU g(-1) dry FSM up to 133 d. We conclude that E. coli O157 can persist and may even grow in feedlot soils, over a wide range of water and manure contents. Further investigations are needed to determine if these variables can be manipulated to reduce this pathogen in cattle and the feedlot environment.     

Influence of aerobic and anaerobic conditions on survival of Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium in Luria-Bertani broth, farm-yard manure and slurry

The influence of aerobic and anaerobic conditions on the survival of the enteropathogens Escherichia coli O157:H7 and Salmonella serovar Typhimurium was investigated in microcosms with broth, cattle manure or slurry. These substrates were inoculated with a green fluorescent protein transformed strain of the enteropathogens at 10(7) cells g(-1) dry weight. Survival data was fitted to the Weibull model. The survival curves in aerobic conditions generally showed a concave curvature, while the curvature was convex in anaerobic conditions. The estimated survival times showed that E. coli O157:H7 survived significantly longer under anaerobic than under aerobic conditions. Survival ranged from approximately. 2 weeks for aerobic manure and slurry to more than six months for anaerobic manure at 16 °C. On average, in 56.3% of the samplings, the number of recovered E. coli O157:H7 cells by anaerobic incubation of Petri plates was significantly (p < 0.05) higher in comparison with aerobic incubation. Survival of Salmonella serovar Typhimurium was not different between aerobic and anaerobic storage of LB broth or manure as well as between aerobic and anaerobic incubation of Petri dishes. The importance of changes in microbial community and chemical composition of manure and slurry was distinguished for the survival of E. coli O157:H7 in different oxygen conditions.     

Method to Partition Between Attached and Unattached E. coli in Runoff From Agricultural Lands1

Abstract: Remediation of waters impaired by bacterial indicators is usually dictated by total maximum daily load plans, which are heavily dependent on fate and transport modeling of bacterial indicators. Nonpoint source pollution models are most frequently used to assess bacterial transport to surface waters and most models typically simulate bacterial transport as a dissolved pollutant. Previous studies have found that cells preferentially attach to sediments; however, a variety of techniques have been used to assess attachment including filtration, fractional filtration, and centrifugation. In addition, a variety of chemical and physical dispersion techniques are used to release attached and bioflocculated cells from particulates. Here we developed and validated an easy‐to‐replicate laboratory procedure for separation of unattached from attached E. coli which will also identify particle sizes to which E. coli preferentially attach. Physical and chemical dispersion techniques were evaluated and a combined hand shaker treatment for 10 min followed by dilutions in 1,000 mg/l of Tween 85 significantly increased total E. coli concentrations by 31% when compared with a control. In order to separate unattached from attached fractions, two commonly used techniques, fractional filtration, and centrifugation were combined. The filtration and centrifugation treatments did not reduce E. coli concentrations when compared with a control (p > 0.05), indicating that damage was not inflicted upon the E. coli cells during the separation procedure.     

Colonies of Cliff Swallows on Highway Bridges: A Source of Escherichia coli in Surface Waters1

Sejkora, Patrick, Mary Jo Kirisits, and Michael Barrett, 2011. Colonies of Cliff Swallows on Highway Bridges: A Source of Escherichia coli in Surface Waters. Journal of the American Water Resources Association (JAWRA) 47(6):1275–1284. DOI: 10.1111/j.1752‐1688.2011.00566.x Abstract: Animals, such as birds, are a source of fecal indicator bacteria and pathogens in the environment. Our objective was to determine whether a colony of cliff swallows nesting underneath a bridge would yield a measurable increase in fecal indicator bacteria (specifically Escherichia coli) in the underlying creek. When the swallows were absent, dry‐weather concentrations of E. coli upstream and downstream of the bridge (in Austin, Texas) were below the Texas contact recreation criteria. When the swallows were present, dry‐weather geometric‐mean E. coli concentrations increased significantly from upstream (43 most probable number [MPN]/100 ml) to downstream (106 MPN/100 ml) of the bridge. One exceedance and one near‐exceedance of the Texas single‐sample contact recreation criterion were observed during the swallows’ nesting phase. When the swallows were present, the downstream E. coli geometric‐mean concentration in storm events (875 MPN/100 ml) was significantly higher than the upstream concentration (356 MPN/100 ml), suggesting that runoff flushes swallow feces from the ground into the creek. Although the loading of E. coli from cliff swallows nesting under bridges can be significant (e.g., dry‐weather loading of 3.1 × 10⁸ MPN/day/nest), the zoonotic potential of the cliff swallow must be examined to determine the risk to human health from contact recreation in waters contaminated with cliff swallow feces.     

Spatial and temporal drivers of zoonotic pathogen contamination of an agricultural watershed

In regions where animal agriculture is prominent, such as southern Alberta, higher rates of gastrointestinal illness have been reported when compared with nonagricultural regions. This difference in the rate of illness is thought to be a result of increased zoonotic pathogen exposure through environmental sources such as water. In this study, temporal and spatial factors associated with bacterial pathogen contamination of the Oldman River, which transverses this region, were analyzed using classification and regression tree analysis. Significantly higher levels of fecal indicators; more frequent isolations of Campylobacter spp., Escherichia coli O157:H7, and Salmonella enterica spp.; and higher rates of detection of pig-specific Bacteroides markers occurred at downstream sites than at upstream sites, suggesting additive stream inputs. Fecal indicator densities were also significantly higher when any one of these three bacterial pathogens was present and where there were higher total animal manure units; however, occasionally pathogens were present when fecal indicator levels were low or undetectable. Overall, Salmonella spp., Campylobacter spp., and E. coli O157:H7 presence was associated with season, animal manure units, and total rainfall on the day of sampling and 3 d in advance of sampling. Several of the environmental variables analyzed in this study appear to influence pathogen prevalence and therefore may be useful in predicting water quality and safety and in the improvement of watershed management practices in this and other agricultural regions.     

Predicting Streambed Sediment and Water Column Escherichia coli Levels at Watershed Scale

A sub‐model for the Soil and Water Assessment Tool (SWAT) is developed to predict Escherichia coli levels in the streambed sediment as well as in the water column. New formulations to estimate the levels of E. coli in streambed sediment and the water column are derived. These equations include calculations of E. coli resuspension from the streambed sediment to the water column, E. coli deposition from the water column to the streambed sediment, E. coli growth in the streambed sediment and the water column, and instream E. coli routing. These formulations were programmed in FORTRAN and integrated into SWAT. The modified SWAT model was applied to Squaw Creek Watershed, Iowa, to predict E. coli levels in the stream. Escherichia coli concentrations in the streambed sediment and the water column were monitored extensively in this watershed, and observations were used to verify the model predictions. The model proposed here can predict E. coli concentrations in streambed sediment as well as in the water column. Approximately 58% of the predictions of E. coli levels in the bed sediment were within 1 order of magnitude from the observed value, and in the water column 83% of the predictions of E. coli levels were within 1 order of magnitude. Results suggest that the proposed model will help predictions of instream bacterial contamination.     

Persistence and Microbial Source Tracking of Escherichia coli at a Swimming Beach at Lake of the Ozarks State Park,Missouri

The Missouri Department of Natural Resources (MDNR) has closed or posted advisories at public beaches at Lake of the Ozarks State Park in Missouri because of Escherichia coli (E. coli) concentration exceedances in recent years. Spatial and temporal patterns of E. coli concentrations, microbial source tracking, novel sampling techniques, and beach‐use patterns were studied during the 2012 recreational season to identify possible sources, origins, and occurrence of E. coli contamination at Grand Glaize Beach (GGB). Results indicate an important source of E. coli contamination at GGB was E. coli released into the water column by bathers resuspending avian‐contaminated sediments, especially during high‐use days early in the recreational season. Escherichia coli concentrations in water, sediment, and resuspended sediment samples all decreased throughout the recreational season likely because of decreasing lake levels resulting in sampling locations receding away from the initial spring shoreline as well as natural decay and physical transport out of the cove. Weekly MDNR beach monitoring, based solely on E. coli concentrations, at GGB during this study inaccurately predicted E. coli exceedances, especially on weekends and holidays. Interestingly, E. coli of human origin were measured at concentrations indicative of raw sewage in runoff from an excavation of a nearby abandoned septic tank that had not been used for nearly two years.     

Incidence and persistence of zoonotic bacterial and protozoan pathogens in a beef cattle feedlot runoff control vegetative treatment system

Determining the survival of zoonotic pathogens in livestock manure and runoff is critical for understanding the environmental and public health risks associated with these wastes. The occurrence and persistence of the bacterial pathogens Escherichia coli O157:H7 and Campylobacter spp. in a passive beef cattle feedlot runoff control-vegetative treatment system were examined over a 26-mo period. Incidence of the protozoans Cryptosporidium spp. and Giardia spp. was also assessed. The control system utilizes a shallow basin to collect liquid runoff and accumulate eroded solids from the pen surfaces; when an adequate liquid volume is attained, the liquid is discharged from the basin onto a 4.5-ha vegetative treatment area (VTA) of bromegrass which is harvested as hay. Basin discharge transported E. coli O157, Campylobacter spp., and generic E. coli into the VTA soil, but without additional discharge from the basin, the pathogen prevalences decreased over time. Similarly, the VTA soil concentrations of generic E. coli initially decreased rapidly, but lower residual populations persisted. Isolation of Cryptosporidium oocysts and Giardia cysts from VTA samples was infrequent, indicating differences in sedimentation and/or transport in comparison to bacteria. Isolation of generic E. coli from freshly cut hay from VTA regions that received basin discharge (12 of 30 vs. 1 of 30 control samples) provided evidence for the risk of contamination; however, neither E. coli O157 or Campylobacter spp. were recovered from the hay following baling. This work indicates that the runoff control system is effective for reducing environmental risk by containing and removing pathogens from feedlot runoff.     

Modeling of Escherichia coli Fluxes on a Catchment and the Impact on Coastal Water and Shellfish Quality1

Bougeard, Morgane, Jean‐Claude Le Saux, Nicolas Pérenne, Claire Baffaut, Marc Robin, and Monique Pommepuy, 2011. Modeling of Escherichia coli Fluxes on a Catchment and the Impact on Coastal Water and Shellfish Quality. Journal of the American Water Resources Association (JAWRA) 1‐17. DOI: 10.1111/j.1752‐1688.2011.00520.x Abstract: The simulation of the impact of Escherichia coli loads from watersheds is of great interest for assessing estuarine water quality, especially in areas with shellfish aquaculture or bathing activities. For this purpose, this study investigates a model association based on the Soil and Water Assessment Tool (SWAT) coupled with a hydrodynamic model (MARS 2D; IFREMER). Application was performed on the catchment and estuary of Daoulas area (France). The daily E. coli fluxes simulated by SWAT are taken as an input in the MARS 2D model to calculate E. coli concentrations in estuarine water and shellfish. Model validation is based on comparison of frequencies: a strong relationship was found between calculated and measured E. coli concentrations for river quality (r² = 0.99) and shellfish quality (r² = 0.89). The important influence of agricultural practices and rainfall events on the rapid and large fluctuations in E. coli fluxes from the watershed (reaching three orders of magnitude in <24 hours) is one main result of the study. Response time in terms of seawater quality degradation ranges from one to two days after any important rainfall event (greater than 10 mm/day) and the time for estuary to recover good water quality also mainly depends on the duration of the rainfall. In the estuary, three effects (rainfall, tidal dilution, and manure spreading) have been identified as important influences.     

Population Dynamics of Escherichia coli in Surface Water1

Bucci, Vanni, Marin Vuli?, Xiaodan Ruan, and Ferdi L. Hellweger, 2011. Population Dynamics of Escherichia coli in Surface Water. Journal of the American Water Resources Association (JAWRA) 47(3):611‐619. DOI: 10.1111/j.1752‐1688.2011.00528.x Abstract: Present models of the fate of enteric bacteria in surface water usually assume simple first‐order decay. However, observed decay kinetics are typically biphasic and there is increasing evidence for growth in the ambient environment. These features are similar to the stationary phase of long‐term batch cultures, where the dynamics are the result of repeated cycles of population takeovers by Growth Advantage in Stationary Phase (GASP) mutants able to scavenge nutrients released from dead cells. We investigated the possible role of this mechanism (i.e., growth of an adapted subpopulation) in surface water using laboratory experiments with various strains of Escherichia coli in different types of water. Addition of ampicillin (which only kills dividing cells) to these cultures caused a decrease in the total cell density, demonstrating the presence of a growing subpopulation. Furthermore, long‐term survivors from surface water cultures outcompete naïve cells and take over the population in surface water. This growth advantage is heritable (i.e., aged strains were isolated from a single cell) and consistent with the GASP mechanism, although the responsible loci were not identified. Our results show that E. coli populations in surface water are dynamic and consist of dying naïve cells and growing surface water adapted cells. These results can explain the apparent biphasic decay pattern observed in laboratory experiments. This mechanism may also be important in the ambient environment.     

RESPONSE OF BACTERIA TO THE PRESENCE OF CARBARYL IN WATER. I. RESPONSE OF BACTERIAL INDICATORS VS. RESPONSE OF PATHOGENIC BACTERIAL SPECIES1

ABSTRACT: Escherichia coli, Streptococcus faecalis, Salmonella typhimurium, and Staphylococcus aureus were tested by plate counts and Warburg respirometry to determine effects of the presence of carbaryl in water. E. coli and S. typhimurium were initially stimulated in the presence of the insecticide whereas S. faecalis and S. aureus were inhibited. Over a period of days, all bacteria died off more rapidly in the presence of carbaryl. The implications of these data in regards to bacteriological testing of water quality are discussed.     

Estimating Potential E. coli Sources in a Watershed Using Spatially Explicit Modeling Techniques1

Riebschleager, K.J., R. Karthikeyan, R. Srinivasan, and K. McKee, 2012. Estimating Potential E. coli Sources in a Watershed Using Spatially Explicit Modeling Techniques. Journal of the American Water Resources Association (JAWRA) 48(4): 745‐761. DOI: 10.1111/j.1752‐1688.2012.00649.x Abstract: The Spatially Explicit Load Enrichment Calculation Tool (SELECT) was automated to characterize waste and the associated pathogens from various sources within a mixed land use watershed. Potential Escherichia coli loads in Lake Granbury watershed were estimated using spatially variable governing factors, such as land use, soil condition, and distance to streams. A new approach for characterizing E. coli loads resulting from malfunctioning on‐site wastewater treatment systems (OWTSs) was incorporated into SELECT along with the Pollutant Connectivity Factor (PCF) module. The PCF component was applied to identify areas contributing E. coli loads during runoff events by incorporating the influence of potential E. coli loading, runoff potential, and travel distance to waterbodies. Simulation results indicated livestock and wildlife are potential E. coli contributing sources in the watershed. The areas in which these sources are potentially contributing are not currently monitored for E. coli. The bacterial water quality violations seen around Lake Granbury are most likely the result of malfunctioning OWTSs and pet wastes. SELECT results demonstrate the need to evaluate each contributing source separately to effectively allocate site specific best management practices (BMPs) utilizing stakeholder inputs. It also serves as a powerful screening tool for determining areas where detailed investigation is merited.     

RESPONSES OF BACTERIA TO THE PRESENCE OF CARBARYL IN WATER. II. PURE CULTURE VS. MIXED CULTURE RESPONSE1

ABSTRACT: Escherichia coli. Streptococcus faecalis, Salmonella typhimurium, and Staphylococcus aureus were tested in pure culture and in various combined mixed cultures to determine effects of the presence of carbaryl in water. Oxygen uptake of E. coli and S. typhimirium was stimulated by five mg/L carbaryl when mixed together, but no stimulation was observed when either organism was tested alone or with other bacteria. In mixed culture the survival time of S. faecalis was reduced in the presence of five mg/L carbaryl; however, the survival time of both S. aureus and S. typhimurium was increased in the presence of five mg/L carbaryl. Implications of these results are discussed.     

DEOXYGENATION IN NATURAL STREAMS1

ABSTRACT: Deoxygenation process in which carbonaceous BOD is biochemically oxidized to reduced inorganic compounds is of great significance in the oxygen demand of stream waters. The BOD decay rate traditionally determined in a laboratory might not necessarily be the same as estimated for a natural stream. The variation in biochemical activity could occur due to differences in adsorption, dispersion, and biophysical processes taking place in the two environments. The first stage biochemical oxygen demand and benthal oxygen demand that occur simultaneously in a stream, have been considered together to calculate the gross rate of deoxygenation in polluted waters. The available data on deoxygenation measurements has been analyzed in this study to calculate the carbonaceous BOD rate constants in natural streams. The analysis indicates that the dimensionless deoxygenation parameter is a function of the ratio of the Reynolds number and Froude number of flow. The functional relationship between these two parameters essentially follows a straight line on a log-log plot. The applicability of the relationship does not depend upon the volume of the organic load, size, or location of the stream.