Distribution and Fate of Escherichia coli in Lake Michigan Following Contamination with Urban Stormwater and Combined Sewer Overflows

Escherichia coli distribution and persistence in nearshore Lake Michigan were assessed following heavy rains and sanitary sewer overflow (SSO) and combined sewer overflow (CSO) events over a 5-year period, including an 18-day period following 25.4 cm of rainfall in which intensive studies were conducted following multiple CSO and SSO events. E. coli levels in the Milwaukee estuary and harbor following SSO and CSO events ranged from 10⁴ to nearly 10⁵ CFU/100 mL, which were significantly higher (p ≤ 0.05) than levels following rainfall alone. Sites outside of the breakwall but within the contamination plume (e.g., within 2 km of the harbor) were an order of magnitude lower. Locations 2–5 km from the harbor ranged from below detection limits, of < 1 to 5 CFU/100 mL. E. coli levels corrected for dilution based on specific conductivity measurements were lower than what would be expected for loss due to dilution alone, suggesting a combination of die-off and dilution, were responsible for the rapid disappearance of these organisms outside of the harbor. E. coli and fecal coliforms measured concurrently demonstrated that fecal coliforms could be recovered longer than E. coli in the open waters of the lake. E. coli isolated directly from sewage treatment plant influent were found to have a marked increase in antibiotic resistance traits for ten antibiotics commonly used in the human population compared with isolates from two animal sources of fecal pollution. However, E. coli obtained from sewage impacted water (n = 2,513) and from stormwater impacted water (n = 1,465) collected the previous year when there were no sewage overflows, were found to have no significant difference (p < 0.05) in the frequency of resistance when comparing the two conditions. E. coli survival characteristics and population dynamics are most likely influenced by multiple factors in complex systems such as the watershed/estuarine/lake environments of the Great Lakes.     

食品中大肠菌群测定的分析研究

大肠菌群的检测作为食品检验中的重要微生物学指标,已是常规监测中必不可少的一个项目和判断食品安全的一个重要指标.国家先后制定了一系列的大肠菌群检测方法和标准,并经过了多次修改.本文通过对新、老国家标准检测方法,测定的注意事项,以及对大肠菌群超标原因等进行分析研究,以使食品生产企业和经销商对可能受污染的各项环节加强管理,严格清除污染源,切实保证食品大肠菌群的达标,从而保障食品安全.     

Inhibition of Undesirable Gas Production in Tofu

Of 100 gas-producing bacterial isolates, 65% were Gram-negative. Bacterial growth and gas production in commercial tofu was accompanied by acidification, and pH decreases as low as 5.2. Coliform bacteria constituted 15% of the total bacterial flora. Sporeformers and yeast counts remained < 10³ CFU/mL. Pasteurization was effective in lowering the bacterial counts, and retarded gas production. The addition of lactic acid (reducing pH to 5.5) or lactic acid bacteria (2 × 10⁷ CFU/g) to tofu helped reduce gas production by about 50%. Addition of both lactic acid and Lactobacillus plantarum stopped gas production in contaminated tofu.     

Textural analysis of hyperspectral images for improving contaminant detection accuracy

Previous studies demonstrated a hyperspectral imaging system has a potential for poultry fecal contaminant detection by measuring reflectance intensity. The simple image ratio at 565 and 517 nm images with optimal thresholding was able to detect fecal contaminants on broiler carcasses with high accuracy. However, differentiating false positives from real contaminants, especially cecal feces were challenging. Further image processing such as textural analysis in the spatial domain was able to reduce false positive errors. In this study, textural analysis of hyperspectral images was conducted to improve detection accuracy by reducing false positives. Specifically, textural analysis with co-occurrence matrix of hyperspectral images performed well to identify “true” contamination. In addition, co-occurrence matrix textural features including average, variance, entropy, contrast, correlation, moment of poultry hyperspectral images were investigated for selecting optimal features to represent contamination. Image pre-processing with co-occurrence textural analysis, specifically mean of co-occurrence textural feature from the matrix (0° angle and distance equals to one) followed by image ratio was able to improve fecal detection accuracy without additional optical filters that increase cost for system hardware of multispectral imaging system for on-line application. Mention of trade names or commercial products in this article is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the U.S. Department of Agriculture.     

Biomass Recovery Method for Adenosine Triphosphate (ATP) Quantification Following UV Disinfection

A biomass recovery method was developed to monitor UV disinfection efficacy using adenosine triphosphate (ATP). Typically, disinfection monitoring at wastewater treatment facilities (WWTFs) involves quantifying fecal and total coliforms or colony forming units, the results of which take a minimum of 24 h to produce. ATP quantification immediately before and after UV treatment, which takes only minutes, shows little reduction and often an increase in the microbial population since UV irradiation results in cells that are viable (i.e., still producing ATP) but not culturable. To overcome this, our biomass recovery method incorporates an incubation step to encourage life cycling of microbes. Average log reductions in cellular ATP (cATP) were found to be ?0.28 ± 0.19, ?0.011 ± 0.153, ?0.17 ± 0.32, and 0.065 ± 0.074 using direct ATP measurements on UV-treated samples from WWTFs A, B, C, and D, respectively, while those using the recovery method were correspondingly 0.17 ± 0.34, 1.8 ± 0.8, 0.20 ± 0.35, and 0.72 ± 0.26. The response of the biomass recovery-ATP method indicated a significant direct correlation to the microbial population reduction observed in heterotrophic plate count (HPC) and Colilert® methods using both pure Escherichia coli culture and secondary municipal wastewater effluent.     

Rates of experimental microbiological contamination of fish exposed to polluted water

Fish inhabiting fecally polluted bodies of water are often used for human consumption. Such fish can be contaminated by enteric human pathogens and may pose a potential risk to public health. Controlled experiments with 132 fish of 100 g average weight were conducted to evaluate the rate of contamination of various tissues of fish (tilapia hybrids). The fish were exposed to E. coli introduced into the ambient water at concentrations of up to 10⁶ cfu/ml. Additional experiments were conducted with diluted wastewater containing Aeromonas, enterococci, fecal coliform and F+ coliphages. In another experiment poliovirus I was also added. The highest bacterial concentrations were recovered from the digestive tract (DT), some 5–24 h following exposure, with DT levels essentially similar to those in the inoculated water. In the E. coli experiments, geometric mean levels of about 10² cfu/cm² were recovered from the skin, 26 cfu/g in the spleen and 10² cfu/g in the liver. Most of the muscle samples were not contaminated. Greater contamination was not found under conditions of stress such as high organic load, a water temperature of 37°C or low levels of dissolved oxygen.     

Field observations on the occurrence of new indicators of disinfection efficiency

This paper summarizes studies on the presence of acid-fast and yeast organisms in wastewater and water treatment plants and in surface water. These organisms were found to satisfy three of Bonde's criteria for indicator organisms: presence whenever pathogens are likely to be present; resistance at least equal to that of pathogens; and lack of regrowth in the post-treatment environment. This, plus prior data, indicates that these organisms are at least as acceptable as indicators of disinfection efficiency than the coliform group.     

Efficacy of monitoring and empirical predictive modeling at improving public health protection at Chicago beaches

Efforts to improve public health protection in recreational swimming waters have focused on obtaining real-time estimates of water quality. Current monitoring techniques rely on the time-intensive culturing of fecal indicator bacteria (FIB) from water samples, but rapidly changing FIB concentrations result in management errors that lead to the public being exposed to high FIB concentrations (type II error) or beaches being closed despite acceptable water quality (type I error). Empirical predictive models may provide a rapid solution, but their effectiveness at improving health protection has not been adequately assessed. We sought to determine if emerging monitoring approaches could effectively reduce risk of illness exposure by minimizing management errors. We examined four monitoring approaches (inactive, current protocol, a single predictive model for all beaches, and individual models for each beach) with increasing refinement at 14 Chicago beaches using historical monitoring and hydrometeorological data and compared management outcomes using different standards for decision-making. Predictability (R²) of FIB concentration improved with model refinement at all beaches but one. Predictive models did not always reduce the number of management errors and therefore the overall illness burden. Use of a Chicago-specific single-sample standard—rather than the default 235 E. coli CFU/100 ml widely used—together with predictive modeling resulted in the greatest number of open beach days without any increase in public health risk. These results emphasize that emerging monitoring approaches such as empirical models are not equally applicable at all beaches, and combining monitoring approaches may expand beach access.     

Reactivation and growth of non-culturable indicator bacteria in anaerobically digested biosolids after centrifuge dewatering

Recent literature has reported that high concentrations of indicator bacteria such as fecal coliforms (FCs) were measured in anaerobically digested sludges immediately after dewatering even though low concentrations were measured prior to dewatering. This research hypothesized that the indicator bacteria can enter a non-culturable state during digestion, and are reactivated during centrifuge dewatering. Reactivation is defined as restoration of culturability. To examine this hypothesis, a quantitative polymerase chain reaction (qPCR) method was developed to enumerate Escherichia coli, a member of the FC group, during different phases of digestion and dewatering. For thermophilic digestion, the density of E. coli measured by qPCR could be five orders of magnitude greater than the density measured by standard culturing methods (SCMs), which is indicative of non-culturable bacteria. For mesophilic digestion, qPCR enumerated up to about one order of magnitude more E. coli than the SCMs. After centrifuge dewatering, the non-culturable organisms could be reactivated such that they are enumerated by SCMs, and the conditions in the cake allowed rapid growth of FCs and E. coli during cake storage.