The influence of temperature and organic matter on the bactericidal activity of short-chain organic acids on salmonellas

CHRISTINA A. CHERRINGTON, VIVIEN ALLEN AND M. HINTON. 1992. Acetic and lactic acids and BioAdd, a commercial preparation of formic and propionic acid, were tested at a concentration of 0.1% (w/w) at 20, 30, 40 and 50°C and in the presence of organic material for bactericidal activity against Salmonella serotype Kedougou. BioAdd was the most active of the solutions at all temperatures, followed by lactic acid and acetic acid. The presence of horse blood at all four temperatures, and milk and serum at 50°C, did not greatly affect the antibacterial activity of the acids although yeast extract (50°C) provided some protection for the salmonella. Acid activity was related to low pH values although the bactericidal activity of acetic acid with blood and milk was greater than the unadulterated acid even though the pH was 0.4 units higher.     

Antimicrobial films produced from chitosan.

Antimicrobial films were prepared by dissolving chitosan into hydrochloric, formic, acetic, lactic and citric acid solutions. Below 40 degrees C, the counter ions could be classified into two groups based on their effect on zero-shear-rate viscosity in 2% solutions of organic acids. Chloride and citrate produced solutions with much lower viscosities than formate, acetate and lactate. At higher temperatures, these differences vanished, and the activation energies of viscous flow were all similar between 40 and 60 degrees C. Films prepared from these solutions were evaluated in tension for Young's modulus, stress and elongation at yield and break points. Films made from hydrochloric, formic and acetic acids were hard and brittle, whereas those from lactic and citric acids were soft and could be stretched. Good correlation was found between Young's modulus and volume of the counter ion. Film properties are essentially governed by the volume of the counter ion and not by the interactions between this counter ion and the macromolecule. Results suggest that acetate has the maximum molecular volume above which the film strength decreases very rapidly.     

The influence of temperature and organic matter on the bactericidal activity of short-chain organic acids on salmonellas.

Acetic and lactic acids and BioAdd, a commercial preparation of formic and propionic acid, were tested at a concentration of 0.1% (w/w) at 20, 30, 40 and 50 degrees C and in the presence of organic material for bactericidal activity against Salmonella serotype Kedougou. BioAdd was the most active of the solutions at all temperatures, followed by lactic acid and acetic acid. The presence of horse blood at all four temperatures, and milk and serum at 50 degrees C, did not greatly affect the antibacterial activity of the acids although yeast extract (50 degrees C) provided some protection for the salmonella. Acid activity was related to low pH values although the bactericidal activity of acetic acid with blood and milk was greater than the unadulterated acid even though the pH was 0.4 units higher.     

Biofilm reduction by a new burn gel that targets nociception

AIMS: To compare the ability of an amorphous first aid topical gel containing vinegar, citric acid and EDTA (RescuDerm(TM); RESC) and various derivative formulations to eradicate Pseudomonas aeruginosa (PSEUD) and Staphylococcus epidermidis (STAPH) biofilms. METHODS AND RESULTS: 24-h biofilms prepared using the Minimum Biofilm Elimination Concentration (MBEC) Assay System were exposed for 4 or 24 h to the different gel formulations. Citric acid-free, acetic acid-free or acetic acid-free/sodium acetate-supplemented RESC gels reduced PSEUD and STAPH biofilm formation as effectively as RESC. Substituting the weak organic acids with equivalent concentrations of glacial acetic acid reduced the effectiveness of gel against PSEUD and STAPH biofilms by half, but viable bacterial counts still remained below 4 log(10) CFU/peg. Removal of gelling agent and/or EDTA enhanced efficacy against PSEUD but not STAPH biofilms. An acidified placebo gel formulation generated an only marginal bactericidal effect compared to that of RESC. CONCLUSIONS: RESC is a promising new antimicrobial agent. Its weak organic acid content, rather than merely acidic pH, mediates its considerable in vitro bactericidal efficacy against bacterial biofilms. SIGNIFICANCE AND IMPACT OF THE STUDY: These data, taken together with the observation that RescuDerm possesses broad in vitro bactericidal activity against other pathogen species, suggest the potential usefulness of this product for controlling biofilm formation on a variety of cutaneous traumatic and surgical wounds.     

Inactivation of Escherichia coli and Listeria monocytogenes on iceberg lettuce by dip wash treatments with organic acids

AIMS: To study and compare the efficacy of organic acids and chlorine dipping in inactivation of Escherichia coli and Listeria monocytogenes on fresh-cut iceberg lettuce. METHODS AND RESULTS: Fresh-cut iceberg lettuce leaves were inoculated with E. coli or L. monocytogenes. After inoculation, samples were stored at 4 degrees C for 24 h and dipped in organic acid or chlorine solutions for 2 and 5 min. E. coli and L. monocytogenes were enumerated on selective media. Treatment of fresh-cut iceberg lettuce with chlorine solution caused 1.0 and 2.0 log(10) CFU g(-1) reductions in the number of L. monocytogenes and E. coli, respectively. Maximum reduction for E. coli (about 2.0 log(10) CFU g(-1)) was obtained for samples dipped in lactic or citric acids while maximum reduction for L. monocytogenes (about 1.5 log(10) CFU g(-1)) was attained for samples dipped in lactic acid. CONCLUSIONS: Dipping of iceberg lettuce in 0.5% citric acid or 0.5% lactic acid solution for 2 min could be as effective as chlorine for reducing microbial populations on fresh-cut iceberg lettuce. SIGNIFICANCE AND IMPACT OF THE STUDY: Dipping in solutions containing organic acids is shown to be effective to reduce E. coli and L. monocytogenes on fresh-cut iceberg lettuce.     

OBSERVATIONS ON THE EFFECT OF ADDED HYPOCHLORITE ON THE NUMBERS AND KINDS OF BACTERIA IN MILK

SUMMARY: In an investigation of the effect of added hypochlorite on the bacterial flora and bacterial numbers of milk no bactericidal action was apparent within 15 min of adding the disinfectant unless at least 500 p/m available chlorine had been added. After 24 hr storage at atmospheric temperature a reduction in numbers was found with 10 p/m available chlorine and this became more pronounced with increased concentrations. Micrococci seemed more susceptible to the bactericidal effect than streptococci, which formed, after 24 hr storage, the predominant residual flora where 250 and 500 p/m available chlorine had been added. Acid production by lactic streptococci was retarded at concentrations greater than 50 p/m available chlorine in a 5 hr incubation period at 30°, but after a 24 hr incubation the decrease in acid production was only apparent when 500 or 1,000 p/m available chlorine was present. Little effect on acid production by lactic streptococci was found when less than 50 p/m available chlorine had been added. The residual chlorine content of rinses of equipment sterilized in hypochlorite solutions was negligible.     

Mechanisms of killing of Bacillus subtilis spores by hypochlorite and chlorine dioxide

AIMS: To determine the mechanisms of Bacillus subtilis spore killing by hypochlorite and chlorine dioxide, and its resistance against them. METHODS AND RESULTS: Spores of B. subtilis treated with hypochlorite or chlorine dioxide did not accumulate damage to their DNA, as spores with or without the two major DNA protective alpha/beta-type small, acid soluble spore proteins exhibited similar sensitivity to these chemicals; these agents also did not cause spore mutagenesis and their efficacy in spore killing was not increased by the absence of a major DNA repair pathway. Spore killing by these two chemicals was greatly increased if spores were first chemically decoated or if spores carried a mutation in a gene encoding a protein essential for assembly of many spore coat proteins. Spores prepared at a higher temperature were also much more resistant to these agents. Neither hypochlorite nor chlorine dioxide treatment caused release of the spore core's large depot of dipicolinic acid (DPA), but hypochlorite- and chlorine dioxide-treated spores much more readily released DPA upon a subsequent normally sub-lethal heat treatment than did untreated spores. Hypochlorite-killed spores could not initiate the germination process with either nutrients or a 1 : 1 chelate of Ca2+-DPA, and these spores could not be recovered by lysozyme treatment. Chlorine dioxide-treated spores also did not germinate with Ca2+-DPA and could not be recovered by lysozyme treatment, but did germinate with nutrients. However, while germinated chlorine dioxide-killed spores released DPA and degraded their peptidoglycan cortex, they did not initiate metabolism and many of these germinated spores were dead as determined by a viability stain that discriminates live cells from dead ones on the basis of their permeability properties. CONCLUSIONS: Hypochlorite and chlorine dioxide do not kill B. subtilis spores by DNA damage, and a major factor in spore resistance to these agents appears to be the spore coat. Spore killing by hypochlorite appears to render spores defective in germination, possibly because of severe damage to the spore's inner membrane. While chlorine dioxide-killed spores can undergo the initial steps in spore germination, these germinated spores can go no further in this process probably because of some type of membrane damage. SIGNIFICANCE AND IMPACT OF THE STUDY: These results provide information on the mechanisms of the killing of bacterial spores by hypochlorite and chlorine dioxide.     

The effect of pH, acidulant and temperature on the survival of Yersinia enterocolitica

The survival of Yersinia enterocolitica at sub-optimal temperatures (0–23°C) and growth inhibitory pH values, achieved using a range of acidulants, was investigated. At a given pH, survival was greater the lower the temperature. Sulphuric and citric acids had lower bactericidal activity than acetic and lactic acids and in nearly all cases where the four acids could be compared at the same pH the order of bactericidal activity was acetic > lactic > citric > sulphuric. Attempts to model this behaviour by a negative square root relationship gave good correlation coefficients for plots of the square root of death rate against temperature at different combinations of pH and acidulant but so too did several other functions of death rate. The high coefficient of variation for T ₀ determined from square root plots prevented construction of a combined temperature/pH model similar to that already described for growth.     

Effectiveness of chlorine washing disinfection and effects on the appearance of artichoke and borage

AIM: Optimal conditions for chlorine application to obtain a reasonable decrease in the microbial counts without damaging the appearance of artichoke and borage have been established. METHODS AND RESULTS: The influence of chlorine concentration (0-200 mg l(-1)), pH, addition of organic acids, contact time and presence of protective structures on the microflora and vegetal appearance were studied. When pH was not controlled the effect of chlorine depended on its concentration until the pH increase caused by addition of chlorine reached 8.8. Any further increase in chlorine concentration was nullified by the pH increase. When pH was adjusted to 4.5 with acetic acid, the effectiveness increased with concentration. However, the use of citric acid to control pH caused a sharp decrease in effectiveness at concentration about 250 mg l(-1). The higher effectiveness of chlorine on homogenized plant extracts compared with the whole plant showed the impact of the vegetal structures on the resistance of the microorganisms. For artichoke, a relationship between the effectiveness of chlorine disinfection and its structures was also found. Extended washing times did not affect the total counts. However, in both vegetables, the appearance was affected by the extended contact times. CONCLUSIONS: The solutions rendering the highest microbial reduction with minimum damages were: 50 mg l(-1) free chlorine without pH control for artichoke and 100 mg l(-1) free chlorine at pH 7.0 for borage. SIGNIFICANCE AND IMPACT OF THE STUDY: Specific conditions for chlorine disinfection of artichoke and borage were determined to reduce the microorganisms in minimally processed artichoke and borage without damaging their appearance.     

Sporicidal Properties of Some Halogens

S ummary . Sodium hypochlorite, sodium dichloro iso cyanurate, dichlorodimethyl hydantoin, dibromodimethyl hydantoin and an iodophor have been examined for disinfectant activity against spores of Bacillus cereus. Under the test conditions used sodium hypochlorite was the most effective compound and dibromodimethyl hydantoin was least affected by increase in pH from 6.5 to 8; the activity of the iodophor was unaffected in the pH range 2.3–6.5. Bacillus subtilis spores were much more resistant to the disinfectants than were B. cereus spores. The addition of KBr to solutions of sodium dichloro iso yanurate enhanced its activity at pH 9 but not at pH 7 or 8.
Mixtures of 1.5–4% of NaOH with NaOCl (200 p of available chlorine/m) were much more rapidly sporicidal than either NaOH or NaOCl (pH 9 and above) alone.     

Sporicidal efficacy of pH-adjusted bleach for control of bioburden on production facility surfaces

pH-adjusted bleach was one of the agents used to disinfect contaminated public buildings in the USA following the 2001 bioterrorist attack with Bacillus anthracis spores. A USEPA fact sheet describes the preparation of pH-adjusted bleach by combining diluted sodium hypochlorite (NaOCl) with a controlled amount of 5 % acetic acid. This paper reports a modification of this procedure to qualify the use of pH-adjusted bleach for routine disinfection of cleanroom surfaces in pharmaceutical manufacturing facilities whenever a short contact time is desirable or there is a need for enhanced germicidal or sporicidal activity. Adjustment of pH was obtained reproducibly with either acetic acid or HCl, confirming the feasibility of developing standard procedures for the controlled addition of acid to diluted NaOCl solutions without compromising operator safety and convenience. Efficacy testing using spores from an in-house isolate of Bacillus pumilus confirmed that NaOCl solutions in the pH 5–8 range have much greater sporicidal activity on surfaces than do unadjusted alkaline solutions (pH > 11). With a contact time of 0.5 min, the log₁₀ reduction in spore viable counts was >5.4 for the five representative surfaces tested relative to untreated controls. Solutions of pH-adjusted NaOCl are known to be less stable than unadjusted alkaline solutions. Stability studies were performed by monitoring sporicidal efficacy, level of free available chlorine (FAC), and pH. Testing included several NaOCl concentrations and adjustment to different starting pHs. The efficacy of pH-adjusted solutions persisted in open containers for at least 12 h even though some FAC degradation occurred. In addition, solutions of 0.29 or 0.50 % NaOCl stored at room temperature protected from light retained efficacy for at least 4 weeks, indicating that short-term storage of solutions is possible following pH adjustment. The inorganic chemical degradation of pH-adjusted NaOCl solutions generates chlorate ion, an undesirable by-product. A comparison of chemical stability for 0.12, 0.25, and 0.50 % NaOCl solutions adjusted to different initial pHs indicated that the least chlorate formation occurred with 0.12 % NaOCl.     

Lactobacillus casei metabolic potential to utilize citrate as an energy source in ripening cheese: a bioinformatics approach

AIMS: To identify potential pathways for citrate catabolism by Lactobacillus casei under conditions similar to ripening cheese. METHODS AND RESULTS: A putative citric acid cycle (PCAC) for Lact. casei was generated utilizing the genome sequence, and metabolic flux analyses. Although it was possible to construct a unique PCAC for Lact. casei, its full functionality was unknown. Therefore, the Lact. casei PCAC was evaluated utilizing end-product analyses of citric acid catabolism during growth in modified chemically defined media (mCDM), and Cheddar cheese extract (CCE). Results suggest that under energy source excess and limitation in mCDM this micro-organism produces mainly L-lactic acid and acetic acid, respectively. Both organic acids were produced in CCE. Additional end products include D-lactic acid, acetoin, formic acid, ethanol, and diacetyl. Production of succinic acid, malic acid, and butanendiol was not observed. CONCLUSIONS: Under conditions similar to those present in ripening cheese, citric acid is converted to acetic acid, L/D-lactic acid, acetoin, diacetyl, ethanol, and formic acid. The PCAC suggests that conversion of the citric acid-derived pyruvic acid into acetic acid, instead of lactic acid, may yield two ATPs per molecule of citric acid. Functionality of the PCAC reductive route was not observed. SIGNIFICANCE AND IMPACT OF THE STUDY: This research describes a unique PCAC for Lact. casei. Additionally, it describes the citric acid catabolism end product by this nonstarter lactic acid bacteria during growth, and under conditions similar to those present in ripening cheese. It provides insights on pathways preferably utilized to derive energy in the presence of limiting carbohydrates by this micro-organism.     

Effect of osmotic, alkaline, acid or thermal stresses on the growth and inhibition of Listeria monocytogenes

Five strains of Listeria monocytogenes (a, b, c, d and e) isolated from industrial plants have been subjected to different osmotic, alkaline, acid or thermal stresses. The effects of these treatments on lag-phase (L) and growth rate (mu) of cells in mid-log phase have been followed using an automated optical density monitoring system. Increasing the osmotic pressure by the addition of different amounts of NaCl increased the lag phase and decreased the growth rate. The same phenomena were observed after decreasing the pH of the medium to 5.8, 5.6 or 5.4 by addition of acetic, lactic or hydrochloric acids. The inhibitory effect was: acetic acid > lactic acid > hydrochloric acid. The addition of NaOH to attain pH values of 9.5, 10.0, 10.5 or 11.0 in the medium produced a dramatic increase of the lag phase at pH 10.5 and 11. Growth rates were also decreased while the maximal population increased with high pH values. These effects varied according to strains. Strains d and e were the most resistant to acidic and alkaline stresses, and e was the most affected by the addition of NaCl. A cold shock of 30 min at 0 degree C had limited effects on growth parameters. On the other hand, hyperthermal shocks (55 or 63 degrees C, 30 min) led to similar increased lag phases and to significant increases of the maximal population in all five strains.     

Growth and toxin formation by Clostridium botulinum at low pH values

Spores of Clostridium botulinum were found to initiate growth and to produce toxin in aqueous suspensions of soya protein at pH values as low as 4-2 and in skimmed milk at pH 4.4. Most of the experiments were done with mixed cultures of CI. botulinum types A and B in the presence of two strains of Bacillus subtilis. The role of the latter organism was concluded to be to lower the oxygen content and the E_h of the suspensions. Toxin was produced at pH 4-4 after 4 weeks of incubation at 30^oC when either hydrochloric or citric acids were used as the acidulant and after 12 and 14 weeks when, respectively, lactic and acetic acids were used. Thus, amongst other factors the nature of the acid and not solely the pH value is an important factor in controlling the growth of Cl. botulinum at low pH. Pure cultures of Cl botulinum type A grew at 30^oC under strictly anaerobic conditions and produced toxin at pH 4-3 in the presence of hydrochloric acid.     

Influence of pH on antimicrobial activity of organic acids against rabbit enteropathogenic strain of<Emphasis Type="Italic">Escherichia coli</Emphasis>

Susceptibility of the rabbit enteropathogenic strain Escherichia coli C6 (O128 serogroup) to C6-C14 fatty acids, oleic, citric, lactic and fumaric acid at 5 mg/mL was determined by the plating technique in the near-neutral pH region (pH approximately 6.5), and in a weakly acid and acid environment (pH 5.4 +/- 0.1 and 2.2-2.5, respectively). In the near-neutral pH region caproic and caprylic acid reduced the concentration of viable cells by 3 and 6 orders, respectively. At lower pH the bactericidal effect of caproic acid remained similar, but caprylic acid decreased the concentration of viable cells to < 100/mL. The bactericidal activity of capric acid was low at pH 6.5 but increased at pH 5.3. High environmental acidity was intrinsically bactericidal and at very low pH the effects of fatty acids were thus less pronounced. Citric acid reduced the counts of viable cells to 1/10. Antimicrobial activity of other acids examined was marginal or absent. Medium-chain fatty acids, caprylic and, to a lesser extent, also caproic and capric acid were better antimicrobials than other organic acids examined; the antimicrobial activity of fatty acids toward the C6 strain was pH-dependent. Beneficial effects of citric, lactic and fumaric acid reported by animal nutritionists are thus probably related to factors other than their direct antimicrobial action.     

Sporicidal Activity of Sodium Hypochlorite at Subzero Temperatures

Sodium hypochlorite was an excellent disinfectant at low temperatures. With the addition of ethylene glycol to prevent freezing, hypochlorite solutions at low free available chlorine concentrations, were effective against Bacillus subtilis var. niger spores from 0 to -40 C. The effectiveness of this decontaminant was influenced by temperature, pH, and concentration, with pH 7.2 the optimum for decontamination at all temperatures and concentrations.     

Inactivation of Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa and Staphylococcus aureus on stainless steel and glass surfaces by neutral electrolysed water

AIM: To ascertain the efficacy of neutral electrolysed water (NEW) in reducing Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Listeria monocytogenes on glass and stainless steel surfaces. Its effectiveness for that purpose is compared with that of a sodium hypochlorite (NaClO) solution with similar pH, oxidation-reduction potential (ORP) and active chlorine content. METHODS AND RESULTS: First, the bactericidal activity of NEW was evaluated over pure cultures (8.5 log CFU ml-1) of the abovementioned strains: all of them were reduced by more than 7 log CFU ml-1 within 5 min of exposure either to NEW (63 mg l-1 active chlorine) or to NaClO solution (62 mg l-1 active chlorine). Then, stainless steel and glass surfaces were inoculated with the same strains and rinsed for 1 min in either NEW, NaClO solution or deionized water (control). In the first two cases, the populations of all the strains decreased by more than 6 log CFU 50 cm-2. No significant difference (P相似文献   

Acidic stress induces the formation of P-bodies, but not stress granules, with mild attenuation of bulk translation in Saccharomyces cerevisiae

The stress response of eukaryotic cells often causes an attenuation of bulk translation activity and the accumulation of non-translating mRNAs into cytoplasmic mRNP (messenger ribonucleoprotein) granules termed cytoplasmic P-bodies (processing bodies) and SGs (stress granules). We examined effects of acidic stress on the formation of mRNP granules compared with other forms of stress such as glucose deprivation and a high Ca2+ level in Saccharomyces cerevisiae. Treatment with lactic acid clearly caused the formation of P-bodies, but not SGs, and also caused an attenuation of translation initiation, albeit to a lesser extent than glucose depletion. P-body formation was also induced by hydrochloric acid and sulfuric acid. However, lactic acid in SD (synthetic dextrose) medium with a pH greater than 3.0, propionic acid and acetic acid did not induce P-body formation. The results of the present study suggest that the assembly of yeast P-bodies can be induced by external conditions with a low pH and the threshold was around pH?2.5. The P-body formation upon acidic stress required Scd6 (suppressor of clathrin deficiency 6), a component of P-bodies, indicating that P-bodies induced by acidic stress have rules of assembly different from those induced by glucose deprivation or high Ca2+ levels.     

The influence of acid on astringency of alum and phenolic compounds

Astringency of aqueous solutions of phenolic compounds (grape seed tannins, tannic acid, catechin and gallic acid) increased upon addition of citric acid, whereas the astringency of alum was reduced. Astringency of alum was decreased equivalently by addition of equi-sour levels of lactic acid, citric acid or hydrochloric acid. The difference between alum and the phenolic compounds is speculated to result from chemical modifications affecting binding of the astringents with oral proteins rather than cognitive differences. Chelation of the aluminum ion in alum by acids reduces its availability for interacting with salivary proteins or epithelial proteins. In contrast, the increased astringency produced upon acidification of phenolic compounds is speculated to result from the pH driven increase in the affinity of the phenols for binding with proteins. These results suggest that alum cannot be used interchangeably with phenolic astringents in psychophysical studies.      

A preliminary assessment of Bacillus anthracis spore inactivation using an electrochemically activated solution (ECASOL)

AIM: To evaluate the efficacy of electrochemically activated solution (ECASOL) in decontaminating Bacillus anthracis Ames and Vollum 1B spores, with and without changing the source water hardness and final ECASOL pH. METHODS AND RESULTS: Five different ECASOL formulations were generated, in which the source water hardness and final ECASOL pH were varied, resulting in cases where significant changes in free available chlorine (FAC) and oxidative-reduction potential (ORP) were observed. B. anthracis Ames and Vollum 1B spores were suspended in the various ECASOL formulations for 30 min, and decontamination efficacy was determined; calcium hypochlorite [5% high-test hypochlorite (HTH)] was used as a positive control. The five different ECASOL formulations yielded mean FAC levels ranging from 305 to 464 ppm, and mean ORP levels ranging from +826 to +1000 mV. Exposure to all the ECASOL formulations and 5% HTH resulted in >or=7.0 log reductions in both B. anthracis Ames and Vollum 1B spores. CONCLUSIONS: The present testing demonstrated that ECASOL with a minimum of c. 300-ppm FAC levels and +800-mV ORP inactivated the B. anthracis spores in suspension, similar to 5% HTH. Significance and Impact of the Study: These results provide information for decontaminating B. anthracis Ames and Vollum 1B spores in suspension using ECASOL.