Erosion and subsequent transport state of Escherichia coli from cowpats

Processes by which fecal bacteria enter overland flow and their transportation state to surface waters are poorly understood, making the effectiveness of measures designed to intercept this pathway, such as vegetated buffer strips, difficult to predict. Freshly made and aged (up to 30 days) cowpats were exposed to simulated rainfall, and samples of the cowpat material and runoff were collected. Escherichia coli in the runoff samples were separated into attached (to particles) and unattached fractions, and the unattached fraction was analyzed to determine if the cells were clumped. Within cowpats, E. coli grew for 6 to 14 days, rather than following a typical logarithmic die-off curve. E. coli numbers in the runoff correlated with numbers inside the cowpat. Most of the E. coli organisms eroded from the cowpats were transported as single cells, and only a small percentage (about 8%) attached to particles. The erosion of E. coli from cowpats and the state in which the cells were transported did not vary with time within a single rainfall event or over time as the cowpats aged and dried out. These findings indicate that cowpats can remain a significant source of E. coli in overland flow for more than 30 days. As well, most of the E. coli organisms eroded from cowpats will occur as readily transportable single cells.     

Removal and inactivation of bacteria during alum treatment of a lake.

Flocculation and removal of bacteria were observed during two separate aluminum sulfate (alum) treatments for removal of phosphorus from a eutrophic recreational lake. In addition, die-off and release of bacteria from alum floc were studied in columns under laboratory conditions. Membrane filtration and spread plates were used to determine concentrations of indicator species and total cultivatable bacteria, respectively. During the alum treatment of the lake, 90% of the fecal coliform (FC) population and ca. 70% of the fecal streptococci population were removed from the water column within 72 h. Numbers of FC in the floc on the lake bottom exceeded 2,400/100 ml at 120 h compared with the pretreatment concentration of 30 FC/100 ml. Inactivation of FC in the floc proceeded at a rate of 200 FC/100 ml per 24 h. In a second alum application to the lake, 95% of the total culturable bacterial population was removed from the water column. In a laboratory column study of survival and release rates, over 90% of an Escherichia coli suspension was concentrated in a floc formed at the bottom. E. coli was not released from the floc. The numbers of and survival of E. coli in the floc suggest the probable concentration of other enteric organisms, including pathogens. Thus, the floc poses a potential human health risk if ingested by swimmers or if others use the lake as a potable water source.     

Numbers and transported state of Escherichia coli in runoff direct from fresh cowpats under simulated rainfall

AIMS: To investigate the number of Escherichia coli in runoff derived directly from fresh cowpats and to determine if the E. coli are attached to dense particles, in flocs or as individual cells. METHODS AND RESULTS: Three cowpats were collected monthly from the same farm for 13 months and the number of E. coli in them estimated. A rainfall simulator was used to generate runoff from the individual cowpats, which was fractioned to determine the transported state of any E. coli present. The number of E. coli in the cowpat runoff was highly variable and was strongly correlated with the number of E. coli in the cowpat. Only a small percentage (approx. 8%) of the E. coli in runoff were attached to dense (>1.3 g ml(-1)) particles and there was no evidence of flocculation of the cells. CONCLUSIONS: Escherichia coli in runoff from cowpats are transported predominantly as individual cells. SIGNIFICANCE AND IMPACT OF THE STUDY: Mitigation strategies to reduce the number of faecal bacteria in overland flow from agricultural land need to be designed to trap single bacterial cells.     

Survival of Escherichia coli O157:H7 in farm water: its role as a vector in the transmission of the organism within herds

AIMS: The study aimed to investigate the survival characteristics of Escherichia coli O157:H7 in farm water (FW), and in sterile distilled municipal water (SDW), stored outdoors under field conditions, with or without the addition of faeces (1% w/v), in a farmyard shed and the laboratory at 15 degrees C. METHODS AND RESULTS: Water samples were inoculated with E. coli O157:H7 at 10(3) and 10(6) ml(-1), and sampled over a 31-day period. In FW stored outdoors in a field, E. coli O157:H7 survived for 14 days at temperatures <15 degrees C, at both inoculation levels, while in the laboratory at 15 degrees C, the organism was still detectable at low levels (<1 log10 cfu ml(-1)) after 31 days. The addition of bovine faeces to water outdoors (1% w/v) resulted in survival for 24 days. In SDW inoculated at 10(6) ml(-1) and stored in the laboratory (15 degrees C), only a 2.5 log reduction was observed after 31 days, while the organism could not be detected after 17 days in the field. Preliminary screening of water samples stored outdoors isolated a bacterium which exhibited antimicrobial activity towards E. coli O157:H7. CONCLUSIONS: The survival of E. coli O157:H7 observed in this study illustrates the potential of farm water to act as a vehicle in the transfer of the organism across a herd. SIGNIFICANCE AND IMPACT OF THE STUDY: The difficulty in extrapolating results from controlled laboratory situations to on-farm conditions is also highlighted in this study.     

Survival of pathogenic bacteria in various freshwater sediments

Four human-associated bacteria, Pseudomonas aeruginosa, Salmonella newport, Escherichia coli, and Klebsiella pneumoniae, were tested for survival in five freshwater sediments. Bacterial survival in continuous-flow chambers was monitored over 14-day periods on sediments ranging from organically rich high-clay fractions to organically poor sandy fractions. Bacterial die-off ranged from 1 to 5 orders of magnitude in sediments. E. coli survived as long as or longer than S. newport. P. aeruginosa and K. pneumoniae tended to survive longer than E. coli. Survival of E. coli and S. newport was greater in sediments containing at least 25% clay. Good reproducibility allowed the development of linear models to describe die-off rates.     

Survival of pathogenic bacteria in various freshwater sediments.

Four human-associated bacteria, Pseudomonas aeruginosa, Salmonella newport, Escherichia coli, and Klebsiella pneumoniae, were tested for survival in five freshwater sediments. Bacterial survival in continuous-flow chambers was monitored over 14-day periods on sediments ranging from organically rich high-clay fractions to organically poor sandy fractions. Bacterial die-off ranged from 1 to 5 orders of magnitude in sediments. E. coli survived as long as or longer than S. newport. P. aeruginosa and K. pneumoniae tended to survive longer than E. coli. Survival of E. coli and S. newport was greater in sediments containing at least 25% clay. Good reproducibility allowed the development of linear models to describe die-off rates.     

Thermal death of Escherichia coli O157:H7 in cattle feeds

AIMS: To determine if the temperatures used in feed manufacture are likely to destroy Escherichia coli O157. METHODS AND RESULTS: Two commercial feeds were ground and inoculated with E. coli O157 cells. The feeds were heated to 50, 55, 60, 65 or 70 degrees C. Heating produced quadratic survivor curves, with rapid initial decreases. The survival characteristics of E. coli O157 differed in the two feeds. The reductions observed in one feed may not have been due to heat alone. There was evidence that indigenous anti-E. coli O157 factor(s) in one feed acted with the heat and contributed to the observed rates of bacterial death. Heating at 70 degrees C for 20 or 120 s resulted in approx. 1.3 and 2.2 log reductions in E. coli O157 numbers respectively. Lesser reductions were observed at lower temperatures. CONCLUSIONS: The time/temperature combinations used in commercial pelleting processes would not effectively kill high numbers of E. coli O157. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study to look at the survival of E. coli O157 strains after heat treatment within concentrated animal feed. The study provides information on the likely risk of E. coli O157 surviving the animal feed manufacturing process.     

Erosion and Subsequent Transport State of Escherichia coli from Cowpats

Processes by which fecal bacteria enter overland flow and their transportation state to surface waters are poorly understood, making the effectiveness of measures designed to intercept this pathway, such as vegetated buffer strips, difficult to predict. Freshly made and aged (up to 30 days) cowpats were exposed to simulated rainfall, and samples of the cowpat material and runoff were collected. Escherichia coli in the runoff samples were separated into attached (to particles) and unattached fractions, and the unattached fraction was analyzed to determine if the cells were clumped. Within cowpats, E. coli grew for 6 to 14 days, rather than following a typical logarithmic die-off curve. E. coli numbers in the runoff correlated with numbers inside the cowpat. Most of the E. coli organisms eroded from the cowpats were transported as single cells, and only a small percentage (about 8%) attached to particles. The erosion of E. coli from cowpats and the state in which the cells were transported did not vary with time within a single rainfall event or over time as the cowpats aged and dried out. These findings indicate that cowpats can remain a significant source of E. coli in overland flow for more than 30 days. As well, most of the E. coli organisms eroded from cowpats will occur as readily transportable single cells.     

Optimizing enrichment conditions for the isolation of Escherichia coli O157 in soils by immunomagnetic separation

AIMS: To compare a range of enrichment broths and enrichment temperatures for the isolation of Escherichia coli O157 by immunomagnetic separation (IMS) from sandy, loam and clay soils. METHODS AND RESULTS: Soils were spiked with cocktails of four atoxigenic strains of E. coli O157 and four strains of commensal E. coli. The organisms were stressed by subjecting soils to cycles of freeze/thawing, followed by drying at 20 degrees C for up to 4 days. Nine enrichment broths were trialled based on buffered peptone water, tryptone soya broths and EC broths supplemented with a range of selective additions. Enrichments were incubated for 6 h and assessed by target recovery after IMS on cefixime tellurite sorbitol MacConkey agar (CTSMAC) incubated at 37 degrees C for 24 h. A comparison of enrichment temperatures (37 and 42 degrees C) was also performed. Buffered peptone water (with or without vancomycin) and tryptone soya broth (with or without novobiocin) gave significant increases in recovery of E. coli O157 compared to others tested. In addition, broths incubated at 42 degrees C were superior to those at 37 degrees C for the recovery of E. coli O157. SIGNIFICANCE AND IMPACT OF THE STUDY: This study showed that sub-lethally damaged E. coli O157 surviving in soil can be sensitive to antimicrobial additions. The choice and concentration of these additions is vitally important to optimize target recovery. Some IMS protocols, established for the isolation of E. coli O157, may be unsuitable for the examination of soil samples.     

The optimization of isolation media used in immunomagnetic separation methods for the detection of Escherichia coli O157 in foods

AIMS: To compare media used in immunomagnetic separation (IMS) techniques for the isolation of Escherichia coli O157 from food. METHODS AND RESULTS: Foods, both naturally contaminated and spiked, with low numbers (< 1 g(-1)) of stressed E. coli O157 were enriched in media based on buffered peptone water (BPW), tryptone soya and EC broths incubated at 30, 37, 40 and 42 degrees C. Following immunomagnetic separation, beads were plated on a range of selective agars. CONCLUSION: BPW supplemented with vancomycin (8 mg l(-1)) incubated at 42 degrees C, followed by IMS and subsequent plating of immunobeads onto cefixime tellurite sorbitol MacConkey agar plus either Rainbow or CHROMagar agars, proved optimum for the recovery of spiked, stressed E. coli O157 in minced beef, cheese, apple juice and pepperoni. The same protocol was optimum for recovery from naturally-contaminated minced beef and cheese. SIGNIFICANCE AND IMPACT OF THE STUDY: The optimum protocol would increase isolation rates of E. coli O157 from foods.     

The effect of starvation stress on the porin protein expression of Escherichia coli in lake water

AIMS: The aim was to identify changes in outer membrane proteins (omps), OmpA, OmpC and OmpF, in Escherichia coli under starvation conditions in lake water microcosms studied at different temperatures. METHODS AND RESULTS: Escherichia coli was incubated in lake water microcosms at a variety of temperatures and the omps studied using quantitative densitometric analysis of protein bands of sodium dodecyl sulphate gels of omp preparations. The amount of OmpF increased over the incubation period relative to that of OmpC whereas the relative abundance of OmpA declined, most notably at 25 and 37 degrees C. This change was linked to changes in peak cell volume as determined by cell measurements. CONCLUSIONS: Major changes to the omps of E. coli accompany the adaptation of the organism to starvation conditions in lake water microcosms. SIGNIFICANCE AND IMPACT OF THE STUDY: Prolonged starvation affects the relative amounts of outer membrane porins. This study furthers the understanding of the role played by changes in the omp composition in E. coli during survival in lake water environments.     

Survival of Escherichia coli and Salmonella spp. in estuarine environments

Survival of Escherichia coli and Salmonella spp. in estuarine waters was compared over a variety of seasonal temperatures during in situ exposure in diffusion chambers. Sublethal stress was measured by both selective-versus-resuscitative enumeration procedures and an electrochemical detection method. E. coli and Salmonella spp. test suspensions, prepared to minimize sublethal injury, were exposed in a shallow tidal creek and at a site 7.1 km further downriver. Bacterial die-off and sublethal stress in filtered estuarine water were inversely related to water temperature. Salmonella spp. populations exhibited significantly less die-off and stress than did E. coli at water temperatures of less than 10 degrees C. Although the most pronounced reductions (ca. 3 log units) in test bacteria occurred during seasonally warm temperatures in the presence of the autochthonous microbiota, 10(2) to 10(4) test cells per ml remained after 2 weeks of exposure to temperatures of greater than 15 degrees C. Reductions in test bacteria were associated with increases in the densities of microflagellates and plaque-forming microorganisms. These studies demonstrated the survival potential of enteric bacteria in estuarine waters and showed that survival was a function of interacting biological and physical factors.     

Salinity and shade preferences result in ovipositional differences between sympatric tiger beetle species

1. Adult tiger beetles of the genus Cicindela often co‐occur within a habitat but larvae do not. Larvae are sedentary and form usually permanent burrows at the site of oviposition where they require 1–3 years for development. 2. To test niche partitioning based on ovipositional preference, the behaviour of two sympatric salt marsh tiger beetles, Cicindela circumpicta and C. togata (Coleoptera: Cicindelidae), were examined. 3. In laboratory studies, female C. circumpicta and C. togata distinguished between experimental salinities, with the former preferring 4 parts per thousand (ppt) and the latter preferring 12 ppt. In the field, C. circumpicta larvae were associated with lower salinities (1–3 ppt) and vegetation along the edges of salt flats while C. togata larvae were found on open salt flats often near halophytes (average salinity = 7.8 ppt). 4. In the field, females chose sites for oviposition in response to shade but not vertical landmarks. In a direct test, 53 of 56 new larval burrows occurred in shaded treatments, three in nonshaded controls. 5. Compared with nonshaded controls, shade increased survival of eggs that were collected in the laboratory and placed in the field. For C. circumpicta eggs, 78% placed in shaded treatments hatched, while significantly fewer (22%) hatched in exposed treatments. For C. togata, 43% of eggs placed in the shade hatched, while no eggs placed in exposed treatments hatched. 6. These results support the hypothesis of niche partitioning between C. circumpicta and C. togata based on ovipositional choice and resulting larval habitat.     

Survival of Escherichia coli and Salmonella spp. in estuarine environments.

Survival of Escherichia coli and Salmonella spp. in estuarine waters was compared over a variety of seasonal temperatures during in situ exposure in diffusion chambers. Sublethal stress was measured by both selective-versus-resuscitative enumeration procedures and an electrochemical detection method. E. coli and Salmonella spp. test suspensions, prepared to minimize sublethal injury, were exposed in a shallow tidal creek and at a site 7.1 km further downriver. Bacterial die-off and sublethal stress in filtered estuarine water were inversely related to water temperature. Salmonella spp. populations exhibited significantly less die-off and stress than did E. coli at water temperatures of less than 10 degrees C. Although the most pronounced reductions (ca. 3 log units) in test bacteria occurred during seasonally warm temperatures in the presence of the autochthonous microbiota, 10(2) to 10(4) test cells per ml remained after 2 weeks of exposure to temperatures of greater than 15 degrees C. Reductions in test bacteria were associated with increases in the densities of microflagellates and plaque-forming microorganisms. These studies demonstrated the survival potential of enteric bacteria in estuarine waters and showed that survival was a function of interacting biological and physical factors.     

Survival of Escherichia coli and Vibrio harveyi in Dead Sea water

Abstract When suspensions of Escherichia coli or the marine luminescent bacterium Vibrio harveyi were mixed with Dead Sea water, the number of viable bacteria decreased by 90% in a time varying from less than 1 h to several hours, depending on the bacterial strain tested. Survival was better at low temperatures, and diluting the Dead Sea water permitted prolonged survival of both coliform bacteria and V. harveyi . The death rate of E. coli in Dead Sea water was comparable to that in water from the Great Salt Lake (Utah). The high concentrations of calcium and magnesium in Dead Sea water, rather than the high total salinity, was identified as the main factor responsible for the rapid die-off. Exposure to direct solar irradiation significantly increased the die-off rate of E. coli in Dead Sea water.
Large numbers of coliform bacteria were recovered from the lake at distances of at least 20 m from a sewage discharge site on the western shore of the Dead Sea.     

Surface decontamination of beef inoculated with Salmonella Typhimurium DT104 or Escherichia coli O157:H7 using dry air in a novel heat treatment apparatus

AIMS: To determine the effectiveness of a novel dry air decontamination apparatus in the deactivation of Salmonella serotype Typhimurium DT104 or Escherichia coli O157:H7 on beef surfaces. METHODS AND RESULTS: A laboratory scale dry air decontamination apparatus, capable of producing repeatable and known heating time-temperature cycles on food surfaces was used in decontamination trials. Beef samples were surface inoculated with 7-8 log10CFU cm(-2) of S. Typhimurium DT104 or E. coli O157:H7 and heated at 60, 75, 90 and 100 degrees C using fast and slow heating rates and subsequently held at these temperatures for up to 600 s. A substantial reduction in pathogen numbers was achieved at higher temperatures (90 and 100 degrees C, 4.18-6.06 log10CFU cm(-2)) using both heating rates, but cell survival at these temperatures was also observed. At the lower temperatures, deactivation was small at 60 degrees C in particular it was less than one log unit after 3 min heating. No significant differences were observed when total reductions in pathogen counts were compared for all the temperature/heat up time combinations tested. During slow heating at 90 degrees C, and both heating rates at 100 degrees C, the pattern of deactivation of S. Typhimurium DT104 or E. coli O157:H7 was triphasic. CONCLUSIONS: This study has shown that heating meat surfaces with dry air can achieve substantial reductions in S. Typhimurium DT104 or E. coli O157:H7. As surface decontamination of beef surfaces with dry air had a negative effect on beef colour and appearance, such a decontamination apparatus would be unsuitable for producing meat for retail sale but it could be used to produce safer meat for use in the catering trade. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides researchers and food processors with data on the dynamic changes in S. Typhimurium DT104 and E. coli O157:H7 counts on intact beef surfaces during heating with dry air under realistic (time-varying) temperature conditions.     

Use of microcosms to determine persistence of Escherichia coli in recreational coastal water and sediment and validation with in situ measurements

AIMS: To determine the persistence of the faecal indicator organism Escherichia coli in recreational coastal water and sediment using laboratory-based microcosms and validation with in situ measurements. METHODS AND RESULTS: Intact sediment cores were taken from three distinct coastal sites. Overlying estuarine water was inoculated with known concentrations of E. coli and decay rates from both overlying water and sediment were determined following enumeration by the membrane filtration method at fixed time intervals over a 28-day period. It was demonstrated that E. coli may persist in coastal sediment for >28 days when incubated at 10 degrees C. Escherichia coli survival was found to have an inverse relationship with temperature in both water and sediment. In general the decay rate for E. coli was greater in water than in sediment. Small particle size and high organic carbon content were found to enhance E. coli survival in coastal sediments in the microcosms. CONCLUSIONS: Results of this microcosm study demonstrated the more prolonged survival of E. coli in coastal sediments compared with overlying water, which may imply an increased risk of exposure because of the possible resuspension of pathogenic micro-organisms during natural turbulence or human recreational activity. SIGNIFICANCE AND IMPACT OF THE STUDY: A more accurate estimate of exposure risk has been described which may subsequently be used in a quantitative microbial risk assessment for recreational coastal waters.     

Rapid enzymatic detection of Escherichia coli contamination in polluted river water

AIMS: The relationship between the rate of beta-D-glucuronidase hydrolysis (GLUase-HR) and the E. coli concentration in rivers differing in the extent of faecal pollution was investigated. It was hypothesized that the rate of GLUase-HR is a better surrogate parameter for E. coli concentrations than estimated numbers of faecal coliforms (FC). METHODS AND RESULTS: The GLUase-HR of the water sample filter residues was determined as the rate of cleavage of 4-methylumbelliferyl-beta-D-glucuronide. FC and E. coli concentrations were enumerated using mFC and Chromocult Coliform agar, respectively. Regression analysis revealed that a 90% variation of the variable log GLUase-HR was directly related to the variable log E. coli concentrations. The observed relationship between the log of the FC count and the log of the GLUase activity could be explained by the hydrolysis activity of the E. coli population, as E. coli is a part of the FC group. CONCLUSION: The data suggest that the log of the GLUase-HR can be used as a surrogate parameter for the log of the E. coli concentrations. SIGNIFICANCE AND IMPACT OF THE STUDY: GLUase-HR determination may provide a rapid alternative technique to estimate E. coli concentrations in freshwaters.     

Survival of Escherichia coli and Salmonella spp. after application of sewage sludge to a Pinus radiata forest

AIMS: This study investigated the survival of Escherichia coli and Salmonella spp. in sewage sludge applied to young and old Pinus radiata forest in Spring and Autumn/Winter. METHODS AND RESULTS: Large numbers of E. coli were present in sludge applied to the forest blocks but Salmonella spp. numbers were low or nondetectable. In the mature stand in Spring, numbers of E. coli returned to back-ground after 3 weeks and die-off was significantly correlated with per cent solids of sludge. E. coli survived longer in mature and young stands in Autumn/Winter where numbers did not significantly decrease until weeks 5 and 13, respectively. Salmonella spp. was detectable in the mature stand until week 4 and in the young stand until week 11 in Autumn/Winter. CONCLUSIONS: Microbial die-off was related to desiccation of the sewage sludge, and was faster in warmer, drier conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: In many countries, environmental and health risks associated with the application of sewage sludge to land are minimized by 'best management practice' guidelines, where risks are managed by restriction of public access to these sites. This study provides supporting evidence that withholding periods of greater than 6 months are sufficient to reduce microbial contaminants to background levels.     

Survival of Escherichia coli K12 in seawater

Abstract The fate of an auxotrophic Escherichia coli K12 strain (NF1830) in coastal water was investigated. The E. coli K12 were enumerated after incubation for varying times in seawater. Incubated in raw seawater at 15 and 20°C, the NF1830 decreased from 10⁶ cfu/ml to below detection within six days of incubation, but when incubated at 7°C it persisted longer. The NF1830 was capable of cell division in sterile seawater. Growth was also shown to occur in raw seawater in the presence of autoclaved sediment. The E. coli K12 decreased in number at a much lower rate when incubated in seawater treated with eukaryotic inhibitors. These findings suggest that the die-off of the auxotrophic E. coli K12 strain seen in the raw seawater was caused by grazing of bacterial predators in the seawater.