A long-term study comparing membrane filtration with Colilert defined substrates in detecting fecal coliforms and Escherichia coli in natural waters

Assessment methods for determining the presence and number of fecal bacteria and Escherichia coli (E. coli) in waters, foodstuffs, sewage effluent, and soils have evolved from multiple tube fermentations (MTF's) to membrane filtrations (MF's) to, most recently, defined substrate technologies (DST's). Mounting evidence indicates Colilert DST (IDEXX, Westbrook, ME) to be a versatile assessment technique for detecting and enumerating E. coli over a range of applications. This study compared the performance of Colilert DST with a confirmed standard MF technique using m-FC broth (Millipore, Bedford, MA) in assessing E. coli in ten different environmental water samples obtained monthly over a 3-year period from the upper Appomattox River, VA. For the duration of the study, E. coli counts measured by Colilert DST were positively correlated (Pearson's correlation coefficient=0.956; slope=0.979; p<0.0001) with E. coli counts measured by confirmed MF procedures. The results of a two-factor ANOVA revealed that Colilert DST counts compared equally to confirmed MF counts by year (p=0.974), by stream sampled (p=1.0), and by season (p=0.696). E. coli counts were significantly lower during cold season months (Dec/Jan/Feb) than during warm season months (Jun/Jul/Aug) for each year contributing to marked variation in sample quality. Counts obtained by Colilert DST compared equally to those obtained by MF across all samples and dates for the three years. Colilert DST presents a laboratory protocol that is simpler to manage, quicker to process, and easier to quantify results than MF. These factors, plus the enhanced precision and versatility of Colilert DST over the span of this three-year study attests to its suitability for testing ambient surface waters.     

Investigation of an Escherichia coli environmental benchmark for waterborne pathogens in agricultural watersheds in Canada

Canada's National Agri-Environmental Standards Initiative sought to develop an environmental benchmark for low-level waterborne pathogen occurrence in agricultural watersheds. A field study collected 902 water samples from 27 sites in four intensive agricultural watersheds across Canada from 2005 to 2007. Four of the sites were selected as reference sites away from livestock and human fecal pollution sources in each watershed. Water samples were analyzed for Campylobacter spp., Salmonella spp., Escherichia coli O157:H7, Cryptosporidium spp., Giardia spp., and the water quality indicator E. coli. The annual mean number of pathogen species was higher at agricultural sites (1.54 ± 0.07 species per water sample) than at reference sites (0.75 ± 0.14 species per water sample). The annual mean concentration of E. coli was also higher at agricultural sites (491 ± 96 colony-forming units [cfu] 100 mL(-1)) than at reference sites (53 ± 18 cfu 100 mL(-1)). The feasibility of adopting existing E. coli water quality guideline values as an environmental benchmark was assessed, but waterborne pathogens were detected at agricultural sites in 80% of water samples with low E. coli concentrations (<100 cfu 100 mL(-1)). Instead, an approach was developed based on using the natural background occurrence of pathogens at reference sites in agricultural watersheds to derive provisional environmental benchmarks for pathogens at agricultural sites. The environmental benchmarks that were derived were found to represent E. coli values lower than geometric mean values typically found in recreational water quality guidelines. Additional research is needed to investigate environmental benchmarks for waterborne pathogens within the context of the "One World, One Health" perspective for protecting human, domestic animal, and wildlife health.     

Escherichia coli loading at or near base flow in a mixed-use watershed

This study analyzed the occurrence of Escherichia coli in a mixed land-use watershed with human, cattle, and wildlife fecal inputs located in a karstic geologic region using synoptic monitoring (samples taken throughout the watershed system) during base-flow conditions. The objective of the study was to evaluate the occurrence of E. coli during base-flow conditions for several months at seven different main channel and nine different tributary sampling sites in the Stock Creek watershed, a 49.3-km(2) basin located in Knoxville, TN. Escherichia coli densities were measured using the Colilert (Defined Substrate Technology) method. The instantaneous loads for E. coli were determined from measured flow rates and E. coli densities, with the highest loading rates observed in the late fall. The study indicated a strong correlation between E. coli load rate (colony-forming units [CFU]/d), 7-d antecedent precipitation, and turbidity. Water quality data, however, also exhibited a spatial dependency; for example, the E. coli load rate was better correlated with turbidity in the slower draining basin tailwater sampling sites than in the faster draining upstream headwater sampling sites. In the headwater sites, the E. coli load rate was better correlated with 7-d antecedent precipitation than turbidity.     

Using zebra mussels to monitor Escherichia coli in environmental waters

Use of the zebra mussel (Dreissena polymorpha) as an indicator of previously elevated bacteria concentrations in a watershed was examined. The ability of the zebra mussel to accumulate and purge Escherichia coli over several days was investigated in both laboratory and field experiments. In laboratory experiments, periodic enumeration of E. coli in mussels that had been exposed to a dilute solution of raw sewage demonstrated that (i) maximum concentrations of E. coli are reached within a few hours of exposure to sewage, (ii) the tissue concentration attained is higher than the concentration in the ambient water, and (iii) the E. coli concentrations take several days to return to preexposure concentrations when mussels are subsequently placed in sterile water. In field experiments conducted in southeast Michigan in the Clinton River watershed, brief increases in E. coli concentrations in the water were accompanied by increases in mussel concentrations of E. coli that lasted 2 or 3 d. The ability of mussels to retain and to concentrate E. coli made it possible to detect E. coli in the environment under conditions that conventional monitoring may often miss. Sampling caged mussels in a river and its tributaries may enable watershed managers to reduce the sampling frequency normally required to identify critical E. coli sources, thereby providing a more cost-effective river monitoring strategy for bacterial contamination.     

LACTOSE NEGATIVE ESCHERICHIA COLI FROM RANGELAND STREAMS: SOURCE,ANTIBIOTIC RESISTANCE,AND COLICINOGENICITY1

ABSTRACT: Lactose-negative Escherichia coil from cattle feces appeared as yellow, atypical colonies on m-FC medium plates with water samples from rangeland streams. The lactose-negative E. coil may impact stream water quality analyses if infrequent samples are collected; are less antibiotic resistant than the lactose-positive E. coili isolated from rangeland streams; and are colicinogenic toward all the laboratory strains of E. coil examined and toward 61 percent of the lactose-positive E. coil rangeland-stream isolates that were tested. This latter result could explain the potentially low degree of antibiotic resistance transfer from lactose-positive to lactose-negative E. coil. In addition, the colicinogenicity of the lactose-negative E. coil may interfere with microbiological water quality analyses that depend upon lactose fernientations with mixed populations of coliforms.     

Evaluating Microbial Indicators of Environmental Condition in Oregon Rivers

Traditional bacterial indicators used in public health to assess water quality and the Biolog system were evaluated to compare their response to biological, chemical, and physical habitat indicators of stream condition both within the state of Oregon and among ecoregion aggregates (Coast Range, Willamette Valley, Cascades, and eastern Oregon). Forty-three randomly selected Oregon river sites were sampled during the summer in 1997 and 1998. The public health indicators included heterotrophic plate counts (HPC), total coliforms (TC), fecal coliforms (FC) and Escherichia coli (EC). Statewide, HPC correlated strongly with physical habitat (elevation, riparian complexity, % canopy presence, and indices of agriculture, pavement, road, pasture, and total disturbance) and chemistry (pH, dissolved O2, specific conductance, acid-neutralizing capacity, dissolved organic carbon, total N, total P, SiO2, and SO4). FC and EC were significantly correlated generally with the river chemistry indicators. TC bacteria significantly correlated with riparian complexity, road disturbance, dissolved O2, and SiO2 and FC. Analyzing the sites by ecoregion, eastern Oregon was characterized by high HPC, FC, EC, nutrient loads, and indices of human disturbance, whereas the Cascades ecoregion had correspondingly low counts of these indicators. The Coast Range and Willamette Valley presented inconsistent indicator patterns that are more difficult to characterize. Attempts to distinguish between ecoregions with the Biolog system were not successful, nor did a statistical pattern emerge between the first five principle components and the other environmental indicators. Our research suggests that some traditional public health microbial indicators may be useful in measuring the environmental condition of lotic systems.     

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.     

Sequence-based source tracking of Escherichia coli based on genetic diversity of beta-glucuronidase

High levels of fecal bacteria are a concern for recreational waters; however, the source of contamination is often unknown. This study investigated whether direct sequencing of a bacterial gene could be utilized for detecting genetic differences between bacterial strains for microbial source tracking. A 525-nucleotide segment of the gene for beta-glucuronidase (uidA) was sequenced in 941 Escherichia coli isolates from the Clinton River-Lake St. Clair watershed, 182 E. coli isolates from human and animal feces, and 34 E. coli isolates from a combined sewer. Environmental isolates exhibited 114 alleles in 11 groups on a genetic tree. Frequency of strains from different genetic groups differed significantly (p < 0.03) between upstream reaches (Bear Creek-Red Run), downstream reaches, and Lake St. Clair beaches. Fecal E. coli uidA sequences exhibited 81 alleles that overlapped with the environmental set. An algorithm to assign alleles to different host sources averaged approximately 75% correct classification with the fecal data set. Using the same algorithm, the percent of environmental isolates assignable to humans decreased significantly between Bear Creek-Red Run (30 +/- 3%) and the beaches (17 +/- 2%) (p < 0.05). Birds accounted for approximately 50% of assignable environmental isolates. For combined sewer isolates, the same algorithm assigned 51% to humans. These experiments demonstrate differences in the frequency of different E. coli strains at different locations in a watershed, and provide a "proof in principle" that sequence-based data can be used for microbial source tracking.     

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.     

Persistence of culturable Escherichia coli fecal contaminants in dairy alpine grassland soils

Our knowledge of Escherichia coli (E. coli) ecology in the field is very limited in the case of dairy alpine grassland soils. Here, our objective was to monitor field survival of E. coli in cow pats and underlying soils in four different alpine pasture units, and to determine whether the soil could constitute an environmental reservoir. E. coli was enumerated by MPN using a selective medium. E. coli survived well in cow pats (10(7) to 10(8) cells g(-1) dry pat), but cow pats disappeared within about 2 mo. In each pasture unit, constant levels of E. coli (10(3) to 10(4) cells g(-1) dry soil) were recovered from all topsoil (0-5 cm) samples regardless of the sampling date, that is, under the snow cover, immediately after snow melting, or during the pasture season (during and after the decomposition of pats). In deeper soil layers below the root zone (5-25 cm), E. coli persistence varied according to soil type, with higher numbers recovered in poorly-drained soils (10(3) to 10(4) cells g(-1) dry soil) than in well-drained soils (< 10(2) cells g(-1) dry soil). A preliminary analysis of 38 partial uidA sequences of E. coli from pat and soils highlighted a cluster containing sequences only found in this work. Overall, this study raises the possibility that fecal E. coli could have formed a naturalized (sub)population, which is now part of the indigenous soil community of alpine pasture grasslands, the soil thus representing an environmental reservoir of E. coli.     

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.     

Locating Escherichia coli contamination in a rural South Carolina watershed

One of the problems associated with the use of ambient water quality standards in surface water regulation is the difficulty of identifying and regulating nonpoint source pollution, making such standards unenforceable, especially at the local level. We used the Escherichia coli indicator to locate the most contaminated reaches in rural South Carolina's Bush River watershed (297 km(2), 186 stream-km). We divided the watershed into 20 smaller reaches and sampled each reach multiple times, but restricted each sampling round to one day. We located four low order creek reaches, representing just nine stream-km, where we observed geometric mean E. coli densities of over 1250 E. coli/100 mL; in each case, the source of the contamination (riparian grazing of cattle) was easily identifiable. On the Bush River itself, we observed a step change in one reach where geometric means increased from 106 E. coli/100 mL to 565 E. coli/100 mL over the reach's 10 km length. In this case, the sources of contamination were not as obvious as in the lower order streams; in this case, more advanced Microbial Source Tracking techniques will be required to identify the sources. Nevertheless, this sampling protocol helped locate polluted reaches and provided decision-makers with reasonable justifications for concrete action in deciding where (or where not) to install conservation practices and where more sophisticated (and expensive) MST techniques were warranted.     

Wildlife identified as major source of Escherichia coli in agriculturally dominated watersheds by BOX A1R-derived genetic fingerprints

The presence of Escherichia coli in recreational and potable waters is a major concern to the general public as elevated levels of E. coli suggest the presence of pathogenic bacteria and viruses. Unfortunately, traditional microbial techniques do not allow specific identification of the source of E. coli. This reduces the ability to target management practices that reduce bacterial contamination. In the Finger Lakes region of western New York, USA, wildlife resides in relatively high densities on watersheds dominated by people and dairy farms, and as a result, the sources of fecal degradation of potable and recreational waters are often unknown. In the Conesus Lake watershed, the sources of microbial contamination were assessed using Rep-PCR molecular tools, a method of amplifying repetitive DNA sequences found throughout the E. coli genome to produce distinct fingerprints for a given ecotype. Molecular fingerprints of E. coli isolated from regional populations of cattle, humans, geese and deer were compared to E. coli isolated from stream water samples. Canonical discriminant function analysis indicated that the DNA fingerprints of the original source group isolates were correctly predicted 90.2% of the time. Since land use in the sub-watersheds was dominated by dairy and cash crop farms, it was expected that the majority of E. coli isolated would be identified as cows; however, an unexpectedly high percentage of isolates were identified as wildlife (geese and deer). Geese were the dominant source of E. coli (44.7-73.7% of the total sources) in four sub-watersheds followed by cows (10.5-21.1%), deer (10.5-18.4%), humans (5.3-12.9%) and unidentifiable sources (0.0-11.8%). Management practices intended to decrease the number of cattle or the amount of manure spread in a sub-watershed were reflected in a decrease of E. coli ecotypes associated with dairy cows.     

Effects of Stock Use and Backpackers on Water Quality in Wilderness in Sequoia and Kings Canyon National Parks, USA

During 2010–2011, a study was conducted in Sequoia and Kings Canyon National Parks (SEKI) to evaluate the influence of pack animals (stock) and backpackers on water quality in wilderness lakes and streams. The study had three main components: (1) a synoptic survey of water quality in wilderness areas of the parks, (2) paired water quality sampling above and below several areas with differing types and amounts of visitor use, and (3) intensive monitoring at six sites to document temporal variations in water quality. Data from the synoptic water quality survey indicated that wilderness lakes and streams are dilute and have low nutrient and Escherichia coli concentrations. The synoptic survey sites were categorized as minimal use, backpacker-use, or mixed use (stock and backpackers), depending on the most prevalent type of use upstream from the sampling locations. Sites with mixed use tended to have higher concentrations of most constituents (including E. coli) than those categorized as minimal-use (P ≤ 0.05); concentrations at backpacker-use sites were intermediate. Data from paired-site sampling indicated that E. coli, total coliform, and particulate phosphorus concentrations were greater in streams downstream from mixed-use areas than upstream from those areas (P ≤ 0.05). Paired-site data also indicated few statistically significant differences in nutrient, E. coli, or total coliform concentrations in streams upstream and downstream from backpacker-use areas. The intensive-monitoring data indicated that nutrient and E. coli concentrations normally were low, except during storms, when notable increases in concentrations of E. coli, nutrients, dissolved organic carbon, and turbidity occurred. In summary, results from this study indicate that water quality in SEKI wilderness generally is good, except during storms; and visitor use appears to have a small, but statistically significant influence on stream water quality.     

Identifying fecal sources in a selected catchment reach using multiple source-tracking tools

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 south-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 flowing surface water.     

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.     

A laboratory study of bacteria-facilitated cadmium transport in alluvial gravel aquifer media

Colloids, including bacteria, can dramatically accelerate the transport of heavy metals in ground water. Batch and column experiments were conducted to investigate adsorption of cadmium (Cd) onto Bacillus subtilis spores or Escherichia coli vegetative cells and Cd transport in alluvial gravel aquifer media in the presence of these bacteria. Results of the batch experiments showed that adsorption of Cd onto the bacteria was (i) positively related to solution pH, bacterial concentration, and negative surface charge, but inversely related to Cd concentration and (ii) a rate-limited nonlinear process, but adsorption onto E. coli was much less. For column influent Cd concentrations of about 4 mg/L and bacterial concentrations of > or = 10(5) colony-forming units (cfu)/mL, there was a significant increase in total Cd effluent concentrations. In comparison with controls that did not have bacteria-facilitated transport, Cd traveled 17 to 20 times faster when it traveled with mobile bacteria. However, Cd traveled mostly 2 to 3 times slower during the desorption phase under the influence of bacteria retained in the column. The difference between total and dissolved Cd concentrations was significant during Cd cotransport with B. subtilis spores, but this concentration difference was very small during Cd cotransport with E. coli, suggesting an adsorption-dominant mechanism during Cd cotransport with the spores and the possibility of Cd chelation by the dissolved membrane vesicles secreted from E. coli cell walls. Bacteria-facilitated transport of heavy metals may pose a threat to ground water quality in sites such as landfills and following land disposal of industrial and domestic effluent and sludge.     

Phosphorus, sediment, and Escherichia coli loads in unfenced streams of the Georgia Piedmont, USA

Contamination of unfenced streams with P, sediments, and pathogenic bacteria from cattle (Bos taurus) activity may be affected by the availability of shade and alternative water sources. The objectives of this study were to evaluate water quality in two streams draining tall fescue (Festuca arundinacea Schreb.)-common bermudagrass (Cynodon dactylon L.) pastures with different shade distribution, and to quantify the effects of alternative water sources on stream water quality. For 3 yr, loads of dissolved reactive phosphorus (DRP), total phosphorus (TP), and total suspended solids (TSS) were measured during storm flow, and loads of DRP, TP, TSS, and Escherichia coli were measured every 14 d during base flow. We also used GPS collars to determine amount of time cattle spent in riparian areas. Our results showed that cattle-grazed pastures with unfenced streams contributed significant loads of DRP, TP, TSS, and E. coli to surface waters (p < 0.01). Time spent by cattle in riparian areas as well as storm flow loads of DRP, TP, and TSS were larger (p < 0.08) in the pasture with the smaller amount of nonriparian shade. Water trough availability decreased base flow loads of TSS and E. coli in both streams, and decreased time cattle spent in riparian areas in the pasture with the smaller amount of nonriparian shade (p < 0.08). Our results indicate that possible BMPs to reduce contamination from cattle-grazed pastures would be to develop or encourage nonriparian shade and to provide cattle with alternative water sources away from the stream.     

Watershed vulnerability predictions for the Ozarks using landscape models

Forty-six broad-scale landscape metrics derived from commonly used landscape metrics were used to develop potential indicators of total phosphorus (TP) concentration, total ammonia (TA) concentration, and Escherichia coli bacteria count among 244 sub-watersheds of the Upper White River (Ozark Mountains, USA). Indicator models were developed by correlating field-based water quality measurements and contemporaneous remote-sensing-based ecological metrics using partial least squares (PLS) analyses. The TP PLS model resulted in one significant factor explaining 91% of the variability in surface water TP concentrations. Among the 18 contributing landscape model variables for the TP PLS model, the proportions of a sub-watershed that are barren and in human use were key indicators of water chemistry in the associated sub-watersheds. The increased presence and reduced fragmentation of forested areas are negatively correlated with TP concentrations in associated sub-watersheds, particularly within close proximity to rivers and streams. The TA PLS model resulted in one significant factor explaining 93% of the variability in surface water TA concentrations. The eight contributing landscape model variables for the TA PLS model were among the same forest and urban metrics for the TP model, with a similar spatial gradient trend in relationship to distance from streams and rivers within a sub-watershed. The E. coli PLS model resulted in two significant factors explaining 99.7% of the variability in E. coli cell count. The 17 contributing landscape model variables for the E. coli PLS model were similar to the TP and TA models. The integration of model results demonstrates that forest, riparian, and urban attributes of sub-watersheds affect all three models. The results provide watershed managers in the Ozark Mountains with a broad-scale vulnerability prediction tool, focusing on TP, TA, and E. coli, and are being used to prioritize and evaluate monitoring and restoration efforts in the vicinity of the White River, a major tributary to the Mississippi River and Gulf of Mexico.     

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.