Survival of Campylobacter jejuni and Escherichia coli in groundwater during prolonged starvation at low temperatures

USDA-ARS, AWMRU, Bowling Green, KY 42104, USA.
Journal of Applied Microbiology (Impact Factor: 2.48). 10/2007; 103(3):573-83. DOI: 10.1111/j.1365-2672.2006.03285.x
Source: PubMed


To evaluate the survival of Campylobacter jejuni relative to that of Escherichia coli in groundwater microcosms varying in nutrient composition.
Studies were conducted in groundwater and deionized water incubated for up to 470 days at 4 degrees C. Samples were taken for culturable and total cell counts, nutrient and molecular analysis. Die-off in groundwater microcosms was between 2.5 and 13 times faster for C. jejuni than for E. coli. Campylobacter jejuni had the lowest decay rate and longest culturability in microcosms with higher dissolved organic carbon (4 mg l(-1)). Escherichia coli survival was the greatest when the total dissolved nitrogen (12.0 mg l(-1)) was high. The transition of C. jejuni to the coccoid stage was independent of culturability.
The differences in the duration of survival and response to water nutrient composition between the two organisms suggest that E. coli may be present in the waters much longer and respond to water composition much differently than C. jejuni.
The data from these studies would aid in the evaluation of the utility of E. coli as an indicator of C. jejuni. This study also provided new information about the effect of nutrient composition on C. jejuni viability.

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    • "The VBNC state is defined as a state of dormancy triggered by harsh environmental conditions [15], such as nutrient starvation [16], extreme temperatures [17], and sharp changes in pH or salinity [18]; osmotic stress [19], oxygen availability [20, 21], and damage to or lack of an essential cellular component including DNA; exposure to food preservatives [22] and heavy metals [23, 24]; exposure to white light [25]; activation of lysogenic phages or suicide genes such as sok/hak or autolysins [26]; and decontaminating processes such as pasteurization of milk [27] and chlorination of wastewater [28]. "
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    ABSTRACT: The viable but nonculturable (VBNC) state is a unique survival strategy of many bacteria in the environment in response to adverse environmental conditions. VBNC bacteria cannot be cultured on routine microbiological media, but they remain viable and retain virulence. The VBNC bacteria can be resuscitated when provided with appropriate conditions. A good number of bacteria including many human pathogens have been reported to enter the VBNC state. Though there have been disputes on the existence of VBNC in the past, extensive molecular studies have resolved most of them, and VBNC has been accepted as a distinct survival state. VBNC pathogenic bacteria are considered a threat to public health and food safety due to their nondetectability through conventional food and water testing methods. A number of disease outbreaks have been reported where VBNC bacteria have been implicated as the causative agent. Further molecular and combinatorial research is needed to tackle the threat posed by VBNC bacteria with regard to public health and food safety.
    09/2013; 2013(8113):703813. DOI:10.1155/2013/703813
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    • "Since Xu et al. (38) reported their pioneering study concerning the existence of the viable but non-culturable (VBNC) state over 30 years ago, a large number of papers have been published by researchers worldwide, documenting the VBNC phenomenon in a wide variety of bacteria. Many pathogens, such as Escherichia coli, Vibrio cholerae, Vibrio vulnificus, Shigella sonnei, Shigella flexneri, Campylobacter jejuni, Legionella pneumophila (3, 5, 10, 26, 27, 32, 34, 36), and Salmonella Enteritidis (9) can enter the VBNC state after exposure to adverse environmental conditions such as high/low temperature, osmotic stress, oxidative stress, and nutritional starvation (2, 11, 23, 26, 37). The VBNC state is now generally accepted as a state in which a cell is metabolically active but is incapable of undergoing the cell division necessary to grow and to form a colony on growth media (21, 25, 29). "
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    ABSTRACT: An environmental isolate of Salmonella Enteritidis (SE), grown to the logarithmic phase, rapidly lost culturability by the addition of 3 mM H2O2 to cultures grown in Luria-Bertani (LB) medium; however, some H2O2-treated bacteria regained their culturability in M9 minimal medium, if sodium pyruvate was present at at least 0.3 mM. In addition, most pyruvate analogues, such as bromopyruvate or phenylpyruvate, did not show restoration activity similar to that of pyruvate, except in the case of α-ketobutyrate. Further analysis of the mechanism underlying the resuscitation by pyruvate revealed that although many of the bacteria showed respiratory activity on CTC (5-cyano-2,3-di-(p-tolyl) tetrazolium chloride) reduction with or without pyruvate, the biosynthesis of DNA and protein synthesis were quite different in the presence or absence of pyruvate, i.e., pyruvate endowed the cells with the ability to incorporate much more radio-label into precursors during the resuscitation process. These results suggest that pyruvate is one of the key molecules working in the resuscitation process by taking bacteria from the non-culturable state to the growing and colony-forming state by triggering the synthesis of macromolecules such as DNA and protein.
    Microbes and Environments 04/2013; 28(2). DOI:10.1264/jsme2.ME12174 · 2.23 Impact Factor
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    • "The VBNC state can be triggered by a variety of environmental factors, including: nutrient deprivation (Cook and Bolster, 2007); changes in temperature (Besnard et al., 2002); oxygen levels (Kana et al., 2008); salinity (Asakura et al., 2008); and presence of heavy metals (Ghezzi and Steck, 1999). Cook and Bolster (2007) showed that E. coli and C. jejuni in groundwater transitioned into a VBNC state characterized by changes in morphology and a reduced rate of respiration in response to starvation and decreased temperature. Although pathogens cannot cause infection while in the VBNC state, there is evidence to suggest that their virulence is retained (Oliver, 2010), therefore they remain a public health risk. "
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    ABSTRACT: An understanding of the transport and survival of microbial pathogens (pathogens hereafter) in agricultural settings is needed to assess the risk of pathogen contamination to water and food resources, and to develop control strategies and treatment options. However, many knowledge gaps still remain in predicting the fate and transport of pathogens in runoff water, and then through the shallow vadose zone and groundwater. A number of transport pathways, processes, factors, andmathematicalmodels often are needed to describe pathogen fate in agricultural settings. The level of complexity is dramatically enhanced by soil heterogeneity, as well as by temporal variability in temperature, water inputs, and pathogen sources. There is substantial variability in pathogenmigration pathways, leading to changes in the dominant processes that control pathogen transport over different spatial and temporal scales. For example, intense rainfall events can generate runoff and preferential flow that can rapidly transport pathogens. Pathogens that survive for extended periods of time have a greatly enhanced probability of remaining viable when subjected to such rapid-transport events. Conversely, in dry seasons, pathogen transport depends more strongly on retention at diverse environmental surfaces controlled by a multitude of coupled physical, chemical, and microbiological factors. These interactions are incompletely characterized, leading to a lack of consensus on the proper mathematical framework to model pathogen transport even at the column scale. In addition, little is known about how to quantify transport and survival parameters at the scale of agricultural fields or watersheds. This review summarizes current conceptual and quantitative models for pathogen transport and fate in agricultural settings over a wide range of spatial and temporal scales. The authors also discuss the benefits that can be realized by improved modeling, and potential treatments to mitigate the risk of waterborne disease transmission.
    Critical Reviews in Environmental Science and Technology 01/2013; 43(8):775-893. DOI:10.1080/10643389.2012.710449 · 3.47 Impact Factor
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