Populations Of Indicator Bacteria Research Articles

Strategy to Evaluate Changes in Bacterial Community Profiles and Bacterial Pathogen Load Reduction After Sewage Disinfection.

Sewage effluent discharge is a major source of pathogenic contamination to the environment. The disinfection process is critical for the elimination of pathogens in sewage. In this study, we examined the impact of chlorine disinfection on the total, viable, and culturable populations of indicator bacteria, pathogens, and bacterial communities in two contrasting types of effluents (primarily treated saline and secondarily treated freshwater). Effluents collected bimonthly over 1 year were examined using cultivation, quantitative PCR (qPCR), and 16S rRNA gene amplicon sequencing coupled with or without propidium monoazide (PMA) treatment. The results showed that each type of effluent was characterized by a specific set of representative genera before disinfection. Salinity appeared to be the major abiotic factor associated with the differences in bacterial community compositions. The pathogen analysis pipeline revealed over 20 viable clinically important pathogenic species in the effluents. Although the bacterial communities differed markedly between the two types of effluents before disinfection, the species of pathogens persisting after disinfection were similar, many of them were members of Enterobacter and Vibrio. The relative abundances of all pathogens identified in the amplicon sequences were multiplied by the 16S rRNA gene copy numbers of total bacteria detected by PMA-qPCR to estimate their concentrations. Pathogens remained viable after disinfection reached 8 log10 16S rRNA copies ml−1 effluent. Meanwhile, around 80 % of the populations of three indicator bacteria including Escherichia coli, Enterococcus, and Bacteroidales were viable after disinfection, but over 99 % of the viable E. coli and Enterococcus were in the non-culturable state. We estimated the total pathogen load by adding the concentrations of all viable pathogens and examined their correlations with indicator bacteria of different types, physiological states, and effluents. The results showed that the PMA-qPCR measurement of E. coli is a reliable proxy of bacterial pathogen loads in both types of effluents. The utility of viable indicator bacteria as a biological index to assess the overall bacteriological hazards in effluents is discussed.

Effect of cultivars and irrigation waters on persistence of indicator bacteria on lettuce grown in high tunnel

AbstractEffect of irrigation with groundwater (GW), primary‐treated wastewater (PTWW), secondary‐treated wastewater (STWW), and roof‐collected rainwater (RCR) on the microbial quality of lettuce cultivars “Annapolis,” “Celinet,” and “Coastline” grown in high tunnel was investigated. Lettuce plants were spray irrigated with irrigation waters once a week for 2 weeks and analyzed for indicator and pathogenic bacteria. PTWW irrigation resulted in the highest Escherichia coli recovery on the lettuce plants (4.7 log MPN/g) as compared to irrigation with other three waters on 0 day postirrigation (dpi). Lettuce cultivars affected the bacterial die‐off rate, where E. coli populations reduced the most by 1.5 log MPN/g on “Annapolis” lettuce on 2 dpi. The STWW and RCR irrigation did not significantly influence indicator bacterial populations on lettuce as compared to GW irrigation. The STWW and RCR containing low populations of indicator bacteria may be suitable for lettuce irrigation in Mid‐Atlantic area without affecting its microbial quality.

In stream dynamics of acid mine drainage and untreated sewage

Both acid mine drainage (AMD) and untreated sewage are serious water quality issues faced by poor, rural communities both within the United States and worldwide. In Appalachia, it is common for these water quality issues to coincide. The aim of this study was to investigate the interaction between AMD and untreated sewage in stream. The study area, Murray City, Ohio, was conflicted, blocking construction of a mine water treatment system to maintain sewage ‘treatment’ by the AMD. Data collected over 5 months from Snow Fork Creek in Murray City, Ohio, show that during low flow, high net acidity portions of a water year, the acidity is sufficient to limit fecal indicator bacteria populations below regulatory limits. During high flow, low net acidity portions of the water year, bacterial populations thrive above the regulatory limits until a point where dilution by water of good quality leads to a decrease in fecal indicator bacteria concentrations to near the regulatory limit of 126 CFU/100 mL for primary contact recreation. This study shows that AMD limits the survival of fecal indicator bacteria and suggests that traditional fecal indicator bacteria are insufficient for detecting untreated sewage in acidic streams despite both visual and olfactory evidence.

Bacterial Pathogen Indicator Transport from Livestock Mortality Biopiles.

Biopiles can be used to dispose of slaughterhouse residuals (SLRs); however, the fate of pathogenic bacteria (e.g., pathogenic strains of , ) in these systems is not well understood. The transport of these bacteria in water leaching from the biopile could represent a significant contamination source. This research examined the transport of Enterobacteriaceae and Enterococcaceae indicator bacteria from SLR biopiles. Three biopiles (2.6 m wide by 4.6 m long by 1.8 m high) were formed on soil layers in concrete cells that allowed for real-time monitoring of environmental parameters, hydrologic flux, and indicator bacteria levels in effluent leaching from the piles. In biopile effluent, indicator bacteria populations decreased exponentially following biopile formation. Indicator bacteria loads in effluent constituted <0.01% of the initial indicator bacteria levels in the biopiles, which was attributed to retention, inactivation, and death. Nearly 90% of the total indicator bacteria loads coincided with large precipitation events (>15 mm d). Movement of the indicator bacteria through the biopiles and underlying soil appeared to be consistent with preferential flow phenomena. The populations of the Enterobacteriaceae indicators remained low in conditions of higher soil water content and lower biopile temperatures, whereas the Enterococcaceae indicator appeared to regrow in these conditions. This indicated that bacterial pathogen transport from a biopile could be a concern after the disappearance of conventional bacterial indicators, such as . Management considerations should attempt to divert excess water from entering a biopile, such as locating a biopile under a roof. Unsaturated biopile and soil conditions should be maintained to impede water flow through preferential pathways in the soil underneath a biopile.

The importance of sediment and submerged aquatic vegetation as potential habitats for persistent strains of enterococci in a subtropical watershed

Recent evidence of extended survival of fecal indicator bacteria in sediments and submerged aquatic vegetation (SAV) has raised concerns about using indicator bacteria to reliably detect fecal contamination. We monitored enterococci densities and population structure in water, sediment and SAV simultaneously at sites across a subtropical watershed (Tampa Bay, FL, USA) over one year to determine the extent to which each matrix serves as a potential reservoir of enterococci. SAV harbored significantly higher mean densities of enterococci than sediments, which harbored higher densities than water. Mean enterococci densities were also greater at sites located further upstream in the watershed. The population structure assessed by BOX-PCR genotyping was relatively dissimilar in each sample, although some similarities among samples suggested grouping by location. Strain diversity ranged from very high to negligible, with lowest overall diversity in lake samples taken during the summer. Several strains were highly abundant and cosmopolitan (found across sites, seasons, and matrices) and were identified by 16S rRNA gene sequencing as the Enterococcus species casseliflavus, faecalis, faecium, hirae, and mundtii. The proportional dominance of certain strains suggests the existence of persistent and possibly naturalized indicator bacteria populations that are not directly related to pollution events.

The effects of submerged aquatic vegetation on the persistence of environmental populations of Enterococcus spp.

Enterococcus spp. are utilized worldwide as faecal indicator bacteria, but certain strains exhibit extended survival in environmental habitats and the factors influencing their persistence are poorly understood. We used flowing freshwater mesocosms to explore the effect of submerged aquatic vegetation (SAV) on the persistence of natural enterococci populations from a subtropical lake. The highest mean densities of culturable enterococci over 2 weeks occurred in SAV [8.6 x 10(2) colony-forming units (cfu) per 100 g wet weight], followed by sediments (1.3 x 10(2) cfu per 100 g) and water (18 cfu per 100 ml). However, due to relative differences in the total mass of each substrate in the entire system (water > sediments > SAV), SAV-associated enterococci represented only a minor proportion of the total population. Vegetated mesocosms harboured significantly higher mean cfu per mesocosm and cfu densities in sediments compared with their unvegetated counterparts, suggesting that SAV indirectly facilitates persistence in aquatic habitats. Populations were dominated (> 96%) by a single Enterococcus casseliflavus strain according to BOX-PCR genotyping, which did not change over the 10-month study and strongly suggests bacterial replication in the lake. The presence of such strains in the environment may represent highly competitive, naturalized and reproducing indicator bacteria populations that are not directly related to pollution events.

An Assessment of Sampling Methods and Microbiological Hygiene Indicators for Process Verification in Poultry Slaughterhouses

Studies to determine the appropriateness of the use of populations of indicator bacteria on poultry carcasses for process verification were undertaken in commercial slaughterhouses. Samples were collected from neck skin by excision or from whole carcass rinses and were examined for a range of presumptive process hygiene indicator bacteria. Coefficients of variation were calculated for each bacterial indicator and were significantly lower in excised samples, indicating more reproducible bacterial recovery by this sampling method. Total viable counts of aerobic bacteria, Enterobacteriaceae, and Pseudomonas in samples collected by excision had the lowest coefficients of variation when compared with other indicators and were therefore used for further study. The uncertainties associated with the quantification of each bacterial indicator were calculated and were lowest overall for total viable counts of aerobic bacteria. In general, uncertainty was higher for lower bacterial numbers. Results of microbiological testing on pooled excised neck skin samples were not significantly different from the mean of individually analyzed samples. Bacterial numbers increased by 1 log unit when cultures were stored under chilled conditions typical of those used for transporting samples to external laboratories, but the increases were not significant for Pseudomonas and aerobic bacteria when storage time was less than 17 h. Weak relationships were identified between bacterial indicator numbers and duration of processing, although cleanliness of the processing environment diminished visibly during this time. In the plants visited for this study, there was a poor relationship between presumptive bacterial indicator numbers and process hygiene. Consequently, bacterial analyses for process verification purposes may be of limited value.

Numbers of fecal streptococci and Escherichia coli in fresh and dry cattle, horse, and sheep manure

Livestock are known contributors to stream pollution. Numbers of fecal streptococci and Escherichia coli in manure naturally deposited by livestock in the field are needed for activities related to bacterial source tracking and determining maximum daily bacterial loading of streams. We measured populations of fecal streptococci and E. coli in fresh and dry manure from cattle (Bos taurus L.), horses (Equus caballus L.), and sheep (Ovis aires L.) on farms in southern Idaho. Populations of indicator bacteria in dry manure were often as high as that in fresh manure from horse and sheep. There was a 2 log10 drop in the population of fecal coliform numbers in dry cattle manure from cattle in pastures but not from cattle in pens. Bacterial isolates used in source tracking should include isolates from both fresh and dry manure to better represent the bacterial source loading of streams.

Indicator bacteria and limnological parameters in fish ponds

Abstract The populations of indicator bacteria (mesophilic, coliform and fecal streptococci) together with relevant limnological parameters (temperature, oxygen, BOD and chlorophyll-α) were recorded during a 6 year study of three eutrophic ponds. Ecosystems were manipulated with fish (Cyprinus carpio) in 2-year management cycles: in spring of the first year, the pond was stocked with young fish, and in the fall of the second year it was drained and mature fish were harvested. Fish management had a direct influence on bacterial numbers and on the basic parameters of the systems. A statistical analysis using t-tests found significant differences in oxygen concentration, chlorophyll-α and BOD during both years of fish management. A water temperature model was calculated on an extensive homogenous data set (197 measurements), by fitting a regression curve. During each season, populations of indicator bacteria increased with increasing water temperature, and maximal numbers of bacteria were recorded during the summer months. Correlation analysis (fitted Spearman correlation coefficient) confirmed that temperature had a positive significant effect on population dynamics. The results demonstrate that fish stocking can affect bacterial population, in the sense that during high fish biomass (second years) the numbers of indicator bacteria as well as BOD and phytoplankton were higher. If pond water quality must be optimised for the purposes of drinking or recreation, the planned fish yield may have to be reduced.

Survival, physiological response and recovery of enteric bacteria exposed to a polar marine environment.

Survival, sublethal injury, and recoverability of Escherichia coli, Enterococcus faecalis, Salmonella typhimurium, and Yersinia enterocolitica were investigated by using diffusion chambers over 54 to 56 days of in situ exposure to a polar marine environment (-1.8 degrees C; salinity, 34.5 ppt) at McMurdo Station, Antarctica. Plate counts were used to assess recoverability and injury, whereas direct viable counts (DVCs) and 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) reduction were utilized to determine substrate responsiveness and respiratory activity, respectively. T90 values (times for 10-fold decreases in numbers of recoverable cells) on nonselective medium were ca. 216 to 259 h for E. coli, S. typhimurium, and Y. enterocolitica and 432 h for E. faecalis. Sublethal injury was greater in populations of indicator bacteria than in pathogens. DVCs, CTC reduction, and plate counts indicated progressive increases in viable but nonculturable cells in E. coli, S. typhimurium, and Y. enterocolitica cultures throughout the 54-day exposure. Forty-eight-day exposure of E. coli, S. typhimurium, and Y. enterocolitica resulted in decreased optimal incubation temperatures for colony formation and inability to form colonies at 37 degrees C. The detection of responsive E. coli, S. typhimurium, and Y. enterocolitica by the DVC and CTC methods remained within 1% of inoculum values during 54 days of exposure, indicating some long-term persistence in the viable-but-nonculturable state. Percentages of respiring E. coli and S. typhimurium increased significantly upon addition of nutrients at all temperatures tested, indicating that nutrient availability rather than temperature limited enteric bacterial activity in this very cold environment. Large nutrient inputs to low-temperature marine environments may thus allow for the long-term persistence of enteric bacteria in a nonrecoverable state.

Detection and occurrence of waterborne bacterial and viral pathogens

Detection and occurrence of waterborne bacterial and viral pathogens

Dynamics of indicator bacteria populations in sediment and river water near a combined sewer outfall

Abstract Sediments collected throughout the summer of 1991 from a river bed around a combined sewer outfall were found to have geometric mean faecal coliform and faecal streptococci densities ranging between 102‐104 g‐1 and 101‐102 g‐1 respectively. During the study period, faecal coliform densities in water samples from the same river reach were several logs lower than in the sediment, but still exceeded state guidelines for primary contact in six of nine sample weeks. Bacteria densities in sediment were weakly correlated with particle size and organic content (as represented by loss‐on‐ignition) during steady‐state baseline flows. Correlations were stronger at the outfall site in the two weeks following an overflow event. Multiple regression analyses indicated that organic content was the dominant factor in explaining variability in bacterial densities in post‐overflow sediments. Using a simple combination of sampled physical parameters (flow velocity, river discharge and bed sediment size distribution)...

Effect of waterfowl (Anas platyrhynchos) on indicator bacteria populations in a recreational lake Madison, Wisconsin

A public swimming beach in Madison, wis., experienced intermittent high fecal coliform counts during the late summer and early fall of 1978. Public health officials closed the beach on a number of occasions. A public health survey identified a combination of waterfowl wastes and meteorological events as the explanation for the high bacteria counts. Fecal coliform bacteria were deposited by mallard ducks and multiplied in the beach sands. The bacteria were subsequently transported into the lake and resulted in high fecal coliform counts in the swimming area.

Aquatic indicator bacteria in the high alpine zone.

Selected waters from the high alpine zone within Grand Teton National Park, Wyoming, were analyzed for populations of indicator bacteria during the past three summers to determine the influence of various factors on the quality of these waters. In general the water quality was not significantly influenced by the presence or absence of human visitors but rather by the nature of the biological community through which the streams flowed. A minority of the coliforms that were recovered from all of the sites proved to be fecal coliforms. The fecal streptococci isolated were identified as the species that were found primarily in the fecal material of the native rodent and moose populations. It is concluded that management questions that relate to the carrying capacity of alpine areas should be approached with the aid of other biological parameters along with levels of indicator bacteria in the streams.

Comparative Survival of Indicator Bacteria and Enteric Pathogens in Well Water

The comparative survival of various fecal indicator bacteria and enteric pathogens was studied in a stable well water supply by using membrane chambers. There was more variation in the 29 coliform cultures and they died more rapidly, as a group, than the 20 enterococcus cultures that were examined. The comparative survival of the organisms tested follows: Aeromonas sp. > the shigellae (Shigella flexneri, S. sonnei, and S. dysenteriae) > fecal streptococci > coliforms = some salmonellae (Salmonella enteritidis ser. paratyphi A and D, S. enteritidis ser. typhimurium) > Streptococcus equinus > Vibrio cholerae > Salmonella typhi > Streptococcus bovis > Salmonella enteritidis ser. paratyphi B. S. bovis had a more rapid die-off than did S. equinus, but both had significantly shorter half-lives than the other streptococci. The natural populations of indicator bacteria from human and elk fecal material declined similarly to the pure cultures tested, whereas the die-off of fecal streptococci exceeded the coliforms from bovine fecal material.