Influence of Incubation Conditions on Survival and Acid Tolerance Response of Escherichia coliO157:H7 and Non-O157:H7 Isolates Exposed to Acetic Acid

1998 ◽  
Vol 61 (5) ◽  
pp. 542-546 ◽  
Author(s):  
LESLYE BRUDZINSKI ◽  
MARK A. HARRISON

The increasing frequency of Escherichia coli O157:H7 outbreaks, especially in acidic foods, raises the concern of an acid tolerance response (ATR). Organic acids can be present in processed and preserved foods: shifts in the acid levels of foods due to these acids may allow E. coli to adapt and later tolerate pH levels that would normally inactivate the organism. The effect of temperature and agitation on the ATRs of three E. coli O157:H7 and two non-O157:H7 isolates were determined. Triggered at pH 5.0, the adaptive System of the ATR allowed for up to nearly 1,000-fold enhanced survival of E. coli O157:H7 cells in some cases compared to survival of nonadapted cells at pH 4.0. E. coli O157:H7 isolates revealed greater acid tolerance responses when incubated statically at 32°C, whereas the non-O157:H7 coli isolates exhibited a greater acid tolerance response with orbital agitation at 25°C. The magnitude of response changed over the incubation period.

2004 ◽  
Vol 67 (3) ◽  
pp. 591-595 ◽  
Author(s):  
LARRY R. BEUCHAT ◽  
ALAN J. SCOUTEN

The effects of lactic acid, acetic acid, and acidic calcium sulfate (ACS) on viability and subsequent acid tolerance of three strains of Escherichia coli O157:H7 were determined. Differences in tolerance to acidic environments were observed among strains, but the level of tolerance was not affected by the acidulant to which cells had been exposed. Cells of E. coli O157:H7 adapted to grow on tryptic soy agar acidified to pH 4.5 with ACS were compared to cells grown at pH 7.2 in the absence of ACS for their ability to survive after inoculation into ground beef treated with ACS, as well as untreated beef. The number of ACS-adapted cells recovered from ACS-treated beef was significantly (α = 0.05) higher than the number of control cells recovered from ACS-treated beef during the first 3 days of a 10-day storage period at 4°C, suggesting that ACS-adapted cells might be initially more tolerant than unadapted cells to reduced pH in ACS-treated beef. Regardless of treatment of ground beef with ACS or adaptation of E. coli O157:H7 to ACS before inoculating ground beef, the pathogen survived in high numbers.


1999 ◽  
Vol 62 (5) ◽  
pp. 451-455 ◽  
Author(s):  
JEE-HOON RYU ◽  
YUN DENG ◽  
LARRY R. BEUCHAT

A study was done to determine if various organic acids differ in their inhibitory or lethal activity against acid-adapted and unadapted Escherichia coli O157:H7 cells. E. coli O157:H7 strain E0139, isolated from venison jerky, was grown in tryptic soy broth (TSB) and in TSB supplemented with 1% glucose (TSBG) for 18 h at 37°C, then plated on tryptic soy agar (TSA) acidified with malic, citric, lactic, or acetic acid at pH 5.4, 5.1, 4.8, 4.5, 4.2, and 3.9. Regardless of whether cells were grown in TSB or TSBG, visible colonies were not formed when plated on TSA acidified with acetic, lactic, malic, or citric acids at pH values of ≤5.4, ≤4.5, ≤4.2, or ≤4.2, respectively. Cells not adapted to reduced pH did not form colonies on TSA acidified with lactic acid (pH 3.9) or acetic acid (pH 3.9 and 4.2); however, a portion of acid-adapted cells remained viable on TSA containing lactic acid (pH 3.9) or acetic acid (pH 4.2) and could be recovered in TSB. Inactivation of acid-adapted cells was less than that of unadapted cells in TSB acidified at pH 3.9 with citric, lactic, or acetic acid and at pH 3.4 with malic acid. Significantly (P ≤ 0.05) higher numbers of acid-adapted cells, compared with unadapted cells, were detected 12 h after inoculation of TSB acidified with acetic acid at pH 3.9; in TSB containing lactic acid (pH 3.9), the number of acid-adapted cells was higher than the number of unadapted cells after 5 h. In TSB acidified at pH 3.9 with citric acid or pH 3.4 with malic acid, significantly higher numbers of acid-adapted cells survived. This study shows that organic acids differ in their inhibitory or lethal activity against acid-adapted and unadapted E. coli O157:H7 cells, and acid-adapted cells are more tolerant than unadapted cells when subsequently exposed to reduced pH caused by these acids.


2001 ◽  
Vol 183 (7) ◽  
pp. 2178-2186 ◽  
Author(s):  
Carrie N. Arnold ◽  
Justin McElhanon ◽  
Aaron Lee ◽  
Ryan Leonhart ◽  
Deborah A. Siegele

ABSTRACT The ability of Escherichia coli to survive at low pH is strongly affected by environmental factors, such as composition of the growth medium and growth phase. Exposure to short-chain fatty acids, such as acetate, proprionate, and butyrate, at neutral or nearly neutral pH has also been shown to increase acid survival of E. coli and Salmonella enterica serovar Typhimurium. To investigate the basis for acetate-induced acid tolerance in E. coli O157:H7, genes whose expression was altered by exposure to acetate were identified using gene arrays. The expression of 60 genes was reduced by at least twofold; of these, 48 encode components of the transcription-translation machinery. Expression of 26 genes increased twofold or greater following treatment with acetate. This included six genes whose products are known to be important for survival at low pH. Five of these genes, as well as six other acetate-induced genes, are members of the E. coli RpoS regulon. RpoS, the stress sigma factor, is known to be required for acid tolerance induced by growth at nonlethal low pH or by entry into stationary phase. Disruption of therpoS gene by a transposon insertion mutation also prevented acetate-induced acid tolerance. However, induction of RpoS expression did not appear to be sufficient to activate the acid tolerance response. Treatment with either NaCl or sodium acetate (pH 7.0) increased expression of anrpoS::lacZ fusion protein, but only treatment with acetate increased acid survival.


2002 ◽  
Vol 48 (5) ◽  
pp. 458-462 ◽  
Author(s):  
Chengru Zhu ◽  
Musangu Ngeleka ◽  
Andrew A Potter ◽  
Brenda J Allan

The Fur (ferric uptake regulator) protein is a master regulator of iron metabolism in gram-negative bacteria. In the present study, the effect of a partial deletion of the fur gene on the acid-tolerance response and in vivo virulence of avian Escherichia coli was examined. The fur mutant was unable to trigger the acid-tolerance response as observed in the wild-type parent strain. However, the mutant was as virulent as the wild-type parent strain when tested in 1-day-old chickens by subcutaneous inoculation. These data indicate that the fur gene is involved in the acid-tolerance response but not involved in the virulence of E. coli, as detected by the ability to cause septicemia in our experimental infection.Key words: E. coli, fur, acid-tolerance response.


1999 ◽  
Vol 62 (5) ◽  
pp. 444-450 ◽  
Author(s):  
R. L. BUCHANAN ◽  
S. G. EDELSON ◽  
R. L. MILLER ◽  
G. M. SAPERS

The extent and location of Escherichia coli O157:H7 contamination after intact apples were immersed in cold (2°C) 1% peptone water containing approximately 3 × 107 CFU/ml was assessed using four apple varieties, Golden Delicious, McIntosh, Red Delicious, and Braeburn. Room temperature and refrigerated apples were used to determine the effect of temperature differential on E. coli infiltration. The highest levels of E. coli were associated with the outer core region of the apple, followed by the skin. Apples were subsequently treated by immersing them for 1 min in 2,000 mg/liter sodium hypochlorite, followed by a 1-min tapwater rinse. This treatment reduced pathogen levels by 1- to 3-log cycles but did not eliminate the microorganism, particularly from the outer core region. While E. coli was not detected in the inner core of most apples, warm fruit immersed in cold peptone water occasionally internalized the pathogen. The frequency and extent of internalization of the pathogen was less when cold apples were immersed in cold peptone water. Subsequent dye uptake studies with Golden Delicious apples indicated that approximately 6% of warm apples immersed into a cold dye solution accumulated dye via open channels leading from the blossom end into the core region. However, dye uptake did not occur when the dye solution was warmer than the apple.


2021 ◽  
Vol 17 (2) ◽  
pp. 189-195
Author(s):  
TR Setyawati ◽  
AH Yanti ◽  
R. Kurniatuhadi

The bacterial isolates NrLtF1, NrLtF4, NrLtF5, and NrLtG2 isolated from fecal pellets and gastrointestinal tract of nypha worms (Namalycastis rhodochorde) have cellulolytic, proteolytic activity and produce organic acids. The four isolates have the potency to be developed as probiotics in nypha worm cultivation feed. This study aims to determine the probiotics potency and identify the species of NrLtF1, NrLtF4, NrLtF5, and NrLtG2 isolate based on 16srDNA sequence. The probiotic potency was carried out by the acid tolerance assays on distilled water and 0.3% acid bile media, and the antimicrobial testing against Escherichia coli (MF exp21.12). Bacterial identification was carried out by sequencing of 16sDNA sequence based on GeneBank data. The results showed that the bacterial isolates of NrLtF1, NrLtF4, NrLtF5, and NrLtG2 were able to grow on 0.3% distilled water and acid bile media. However, only the NrLtF4 and NrLtF5 inhibited E. coli (MF exp21.12) with halo zones 30 mm and 18 mm, respectively. Blasting results of the 16srDNA sequences showed that the NrLtF1, NrLtF4, NrLtF5, and NrLtG2 were closely related to Bacillus wiedmannii, Brevibacterium sediminis, Bacillus proteolyticus, and Bacillus paramycoides. The nypha worm bacterial isolates have the potency to be developed as probiotics in nypha worm culture.


2006 ◽  
Vol 69 (8) ◽  
pp. 1865-1869 ◽  
Author(s):  
AAKASH KHURANA ◽  
GEORGE B. AWUAH ◽  
BRADLEY TAYLOR ◽  
ELENA ENACHE

Studies were conducted to evaluate the combined effect of selected acidulants (acetic, citric, malic, and phosphoric acid) and heat on foodborne pathogens (Escherichia coli O157:H7 and Listeria monocytogenes) in pureed green beans. To establish a consistent reference point for comparison, the molar concentrations of the acids remained constant while the acid-to-puree ratio, titratable acidity, and undissociated acid were either measured or calculated for a target acidified green beans at a pH of 3.8, 4.2, and 4.6. The D-values at 149°F were used as the criteria for acid efficacy. Generally, acetic acid (puree, pH 3.8 and 4.2) represented the most effective acid with comparatively low D-values irrespective of the target microorganism. A 10-s heating at 149°F inactivated approximately 106 CFU/ml of E. coli O157:H7 in pureed beans at pH 3.8. The efficacy of acetic acid is likely related to the elevated percent titratable acidity, undissociated acid, and acid-to-puree ratio. The effectiveness (which in this study represents the combined effect of acid and heat) of the remaining acids (citric, malic, and phosphoric) at puree pH values of 3.8 and 4.2 were statistically insignificant (α = 0.05). Surprisingly, acetic acid (puree, pH 4.6) appeared to be the least effective as compared to the other acids tested (citric, malic, and phosphoric) especially on E. coli O157:H7 cells, while L. monocytogenes had a similar resistance to all acids at puree pH 4.6. With the exception of citric acid (pH 3.8), acetic acid (pH 4.6), and malic acid (pH 3.8 and 4.6), which were statistically insignificant (P > 0.05), the D-values for L. monocytogenes were statistically different (P ≤ 0.05) and higher than the D-values for E. coli under similar experimental conditions. A conservative process recommendation (referred to as the “safe harbor” process) was found sufficient and applicable to pureed green beans for the pH range studied.


Pathogens ◽  
2020 ◽  
Vol 9 (10) ◽  
pp. 849
Author(s):  
Vinicius Silva Castro ◽  
Yhan da Silva Mutz ◽  
Denes Kaic Alves Rosario ◽  
Adelino Cunha-Neto ◽  
Eduardo Eustáquio de Souza Figueiredo ◽  
...  

Salmonella and Escherichia coli are the main bacterial species involved in food outbreaks worldwide. Recent reports showed that chemical sanitizers commonly used to control these pathogens could induce antibiotic resistance. Therefore, this study aimed to describe the efficiency of chemical sanitizers and organic acids when inactivating wild and clinical strains of Salmonella and E. coli, targeting a 4-log reduction. To achieve this goal, three methods were applied. (i) Disk-diffusion challenge for organic acids. (ii) Determination of MIC for two acids (acetic and lactic), as well as two sanitizers (quaternary compound and sodium hypochlorite). (iii) The development of inactivation models from the previously defined concentrations. In disk-diffusion, the results indicated that wild strains have higher resistance potential when compared to clinical strains. Regarding the models, quaternary ammonium and lactic acid showed a linear pattern of inactivation, while sodium hypochlorite had a linear pattern with tail dispersion, and acetic acid has Weibull dispersion to E. coli. The concentration to 4-log reduction differed from Salmonella and E. coli in acetic acid and sodium hypochlorite. The use of organic acids is an alternative method for antimicrobial control. Our study indicates the levels of organic acids and sanitizers to be used in the inactivation of emerging foodborne pathogens.


2013 ◽  
Vol 76 (7) ◽  
pp. 1245-1249 ◽  
Author(s):  
F. BREIDT ◽  
K. KAY ◽  
J. COOK ◽  
J. OSBORNE ◽  
B. INGHAM ◽  
...  

A critical factor in ensuring the safety of acidified foods is the establishment of a thermal process that assures the destruction of acid-resistant vegetative pathogenic and spoilage bacteria. For acidified foods such as dressings and mayonnaises with pH values of 3.5 or higher, the high water phase acidity (acetic acid of 1.5 to 2.5% or higher) can contribute to lethality, but there is a lack of data showing how the use of common ingredients such as acetic acid and preservatives, alone or in combination, can result in a 5-log reduction for strains of Escherichia coli O157:H7, Salmonella enterica, and Listeria monocytogenes in the absence of a postpackaging pasteurization step. In this study, we determined the times needed at 10°C to achieve a 5-log reduction of E. coli O157:H7, S. enterica, and L. monocytogenes in pickling brines with a variety of acetic and benzoic acid combinations at pH 3.5 and 3.8. Evaluation of 15 different acid-pH combinations confirmed that strains of E. coli O157:H7 were significantly more acid resistant than strains of S. enterica and L. monocytogenes. Among the acid conditions tested, holding times of 4 days or less could achieve a 5-log reduction for vegetative pathogens at pH 3.5 with 2.5% acetic acid or at pH 3.8 with 2.5% acetic acid containing 0.1% benzoic acid. These data indicate the efficacy of benzoic acid for reducing the time necessary to achieve a 5-log reduction in target pathogens and may be useful for supporting process filings and the determination of critical controls for the manufacture of acidified foods.


1996 ◽  
Vol 59 (5) ◽  
pp. 453-459 ◽  
Author(s):  
PINA M. FRATAMICO ◽  
FRANKIE J. SCHULTZ ◽  
ROBERT C. BENEDICT ◽  
ROBERT L. BUCHANAN ◽  
PETER H. COOKE

Attachment of E. coli O157:H7 and E. coli K12 to beef tenderloin filet, chuck, and adipose tissues was studied. Most attachment occurred within 1 min of incubation; the number of attached organisms depended on the concentration of bacteria in the liquid inoculum. Similar levels of E. coli bound to the three types of beef tissues tested. E. coli O157:H7 was heavily piliated; however, there was no significant difference between levels of bound E. coli O157:H7 and E. coli K12, indicating that these surface structures apparently are not involved in attachment. Scanning electron photomicrographs of meat tissue and of purified collagen suggested that bacteria attached primarily to collagen fibers. Rinsing solutions consisting of 10% trisodium phosphate (TSP), 2% acetic acid (HAc), phosphate-buffered saline (PBS) and combinations of each were tested for effectiveness in reducing the number of attached E. coli. The level of bacteria removed from tenderloin tissue following TSP, HAc, or PBS rinses did not differ considerably. When beef tissues were stored at 4°C for 18 h after the various rinse combinations, TSP rinse treatments reduced the levels of E. coli K12 and O157:H7 attached to adipose tissue up to 3.4 and 2.7 log units, respectively, compared to PBS rinse treatments. Therefore, TSP may be effective for reducing populations of E. coli O157:H7 on beef carcass tissue.


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