[논문]The Effect of Environmental Factors on Phage Stability and Infectivity on Their Host Bacteria: a Case Study for an Escherichia coli Phage (T7), a Listeria Phage (A511), and a Salmonella Phage (Felix O1)

Kim, Kwang-Pyo

The Effect of Environmental Factors on Phage Stability and Infectivity on Their Host Bacteria: a Case Study for an Escherichia coli Phage (T7), a Listeria Phage (A511), and a Salmonella Phage (Felix O1) 원문보기

Food science and biotechnology, v.16 no.3 = no.75, 2007년, pp.398 - 403

Kim, Kwang-Pyo (Food Microbiology Laboratory, Institute of Food Science and Nutrition ETH Zurich)

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The effectiveness of phage biocontrol depends on the activity of bacteriophage in a given environment. In order to investigate the infectivity and the stability of bacteriophages in representative environments, three virulent phages, Listeria phage A511, Salmonella phage Felix O1, and Escherichia coli phage T7, were subjected to different temperatures, pHs and salt concentrations (NaCl). Phage infectivity was also determined in the presence of divalent cations ($Mg^{2+}$ or $Ca^{2+}$). As a result, three phages exhibited a wide range of survival rates under various environments. Phage infectivity was directly correlated with bacterial growth under the applied conditions. One exception was Felix O1 that did not kill Salmonella grown in low pH (4.5). The failure was attributed to defective adsorption of Felix O1. This finding is significant as it provides an explanation for the inefficient phage biocontrol. Therefore, such information is crucial to improve phage biocontrol of pathogens.

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가설 설정

(1-3). Advantages include: (i) specificity, phages intrinsically contain the ability to destroy the host target bacteria. The host range could be at the species or even strain level, which is advantageous because many foods require continuous presence of non-harmful microorganisms; (ii) efficiency, phage treatment in food could be as effective as chemical sanitizers in destroying pathogens (1) and spoilage microorganisms (4, 5); (ⅲ) non-destructive and non-thermal method to keep the freshness of food; (ⅳ) safety, phage itself is merely a nucleic acid molecule covered by protein capsid which might contain some lipids.
1)Bacterial growth was shown by positive (+) or negative (-) determination. Infectivity was determined by taking percent of PFU under the applied condition compare to that in the control.

제안 방법

Phage infectivities at different temperatures (4, 10, 30, 37, and 42℃) were also measured by using BHI 1/ 2 soft agar overlayered plates. Controls were either, 30℃ for A511 and 37℃ for Felix O₁ and T7 for temperature, pH 7.5 for pH, without addition of NaCl for salt treatment, or no addition of divalent cations.
I thank Prof. Martin J. Loessner for the support, and Prof. Markus Schuppler for help to prepare the manuscript.
2), its inability to kill Salmonella is likely to be related to the infection process(es). In an effort to find out a possible explanation for such a feilure by Felix O₁ in low pH,phage adsorption, the first step of phage-host interaction, was tested. When Felix O₁ was incubated with Salmonella cells grown at pH 4.
Phage infectivity determined by measuring either efficiency of plating (EOP) or bacterial lysis in liquid culture Phage infectivity was determined by measuring EOP (in this study, meaning percent PFU in the given condition compare to that in the control) in various media conditions. Exponentially growing bacteria were infected with phage (102-3 PFU/plate) in molten soft agar (0.
This observation could be very important in analyzing the outcomes of phage biocontrol. The basic information obtained in this study will be of great value to deepen the understanding of tfie relevant mechanisms of phage biocontrol.
In this study, I present the data suggesting that the host growth environment could affect the outcomes of phage biocontrol. Three broad host range virulent phages (A511, Felix Ol, and T7), potential candidates for biocontrol agent, were tested for their infectivity and stability under representative environmental conditions (temperature, pH, salts, presence of divalent cations). Bacterial lysis in liquid media and phage adsorption were also determined when necessary.

대상 데이터

Phage survival at various temperatures. Three phages (A511, Felix 01, and T7) were purified by PEG precipitation followed by ultracentrifuge in CsCl gradient and approximately 1011 PFU/mL of each phage was incubated at various temperatures (A, -20℃; B, 4℃; 37℃; D, 42℃). At each designated time point, phage survival was measured by soft agar layer technique.

데이터처리

error bars. The data obtained were analyzed by standard Student's /-test (a = 0.05).

성능/효과

In conclusion, based on our results and previous studies, phages seem to be quite stable particles that were resistant to a wide range of environmental conditions, but at various rates. Therefore, certain phage resistance properties need to be checked individually.
3 log in 10 min. The number of infective Felix 01 and T7 decreased in 10 min by only 0.5 and 1.0 log PFU, respectively, which was statistically significant (pv0.05) (Table 1).

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The Effect of Environmental Factors on Phage Stability and Infectivity on Their Host Bacteria: a Case Study for an Escherichia coli Phage (T7), a Listeria Phage (A511), and a Salmonella Phage (Felix O1) 원문보기

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The Effect of Environmental Factors on Phage Stability and Infectivity on Their Host Bacteria: a Case Study for an Escherichia coli Phage (T7), a Listeria Phage (A511), and a Salmonella Phage (Felix O1) 원문보기

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