Use of biocides for the control of fungal outbreaks in subterranean environments: the case of the Lascaux Cave in France

The Lascaux Cave in France suffered an outbreak of the fungus Fusarium solani in 2001. Biocides were applied for three years to control this outbreak. Four months after the initial biocide application, a new outbreak appeared in the form of black stains that progressively invaded the cave. The black stains on the ceiling and passage banks were so evident by 2007 that they became one of the cave's major problems. Therefore, biocides were used again in 2008. The present study investigated the fungal communities associated with the black stains and the effectiveness of the biocides applied, by using cloning, denaturing gradient gel electrophoresis, and culture-dependent methods. A novel species, Ochroconis lascauxensis, was the most abundant fungus in samples collected between 2007 and 2008, and the biocides applied were not effective in eliminating this fungus; on the contrary, they appeared to increase the fungal diversity. The fungal communities represented in the samples collected in 2010 were quite different from those collected in 2008 and 2009: the major OTUs corresponded to black yeasts belonging to the Herpotrichiellaceae family. The origin and evolution of these microorganisms are probably linked to the intensive biocide treatments and to the anthropogenic changes introduced by cave management.     

Effect of enterocin AS-48 in combination with biocides on planktonic and sessile Listeria monocytogenes

Enterocin AS-48 was tested on a cocktail of Listeria monocytogenes strains in planktonic and sessile states, singly or in combination with biocides benzalkonium chloride, cetrimide, hexadecylpyridinium chloride, didecyldimethylammonium bromide, triclosan, poly-(hexamethylen guanidinium) hydrochloride, chlorhexidine, hexachlorophene, and the commercial sanitizers P3 oxonia and P3 topax 66. Combinations of sub-inhibitory bacteriocin concentrations and biocide concentrations 4 to 10-fold lower than their minimum inhibitory concentrations (MIC) completely inhibited growth of the planktonic listeriae. Inactivation of Listeria in biofilms formed on polystyrene microtiter plates required concentrations of enterocin AS-48 greater than 50 μg/ml, and biocide concentrations ten to 100-fold higher. In combination with enterocin AS-48 (25 or 50 μg/ml), microbial inactivation increased remarkably for all biocides except P3 oxonia and P3 topax 66 solutions. Polystyrene microtiter plates conditioned with enterocin solutions (0.5-25 μg/ml) decreased the adherence and biofilm formation of the L. monocytogenes cell cocktail, avoiding biofilm formation for at least 24 h at a bacteriocin concentration of 25 μg/ml.     

Adaptive tolerance to phenolic biocides in bacteria from organic foods: Effects on antimicrobial susceptibility and tolerance to physical stresses

The aim of the present study was to analyze the effects of step-wise exposure of biocide-sensitive bacteria from organic foods to phenolic biocides triclosan (TC) and hexachlorophene [2,2′-methylenebis(3,4,6-trichlorophenol)] (CF). The analysis included changes in the tolerance to the biocide itself, the tolerance to other biocides, and cross-resistance to clinically important antibiotics. The involvement of efflux mechanisms was also studied as well as the possible implication of modifications in cytoplasmic membrane fluidity in the resistance mechanisms. The influence of biocide tolerance on growth capacity of the adapted strains and on subsequent resistance to other physical stresses has also been analyzed. Repeated exposure of bacteria from organic foods to phenolic biocides resulted in most cases in partially increased tolerance to the same biocide, to dissimilar biocides and other antimicrobial compounds. Nine TC-adapted strains and six CF-adapted strains were able to develop high levels of biocide tolerance, and these were stable in the absence of biocide selective pressure. Most strains adapted to TC and one CF-adapted strain showed significantly higher anisotropy values than their corresponding wildtype strains, suggesting that changes in membrane fluidity could be involved in biocide adaptation. Exposure to gradually increasing concentrations of CF induced a decrease in heat tolerance. Biocide adaptation had no significant effects of gastric acid or bile resistance, suggesting that biocide adaptation should not influence survival in the gastrointestinal tract.     

Heat shock treatments: a new safe approach against lichen growth on outdoor stone surfaces

The control of lichen growth, particularly important in the field of stone conservation of outdoor monuments, largely depends on the use of biocides, that may be dangerous for the users, the environment and the substratum. A new, alternative approach is proposed, which makes the most of a poorly known peculiarity of poikilohydrous organisms: they are thermo-tolerant (up to 65-70 °C) when dry, but thermo-sensitive when wet. The efficacy of thermal treatments (range: 20-55 °C), in parallel to the application of three biocides, was verified in the laboratory with six epi- and endolithic lichens. Chlorophyll a fluorescence emission was checked in treated and nontreated samples of all the species, whereas histochemical observations with a dead cell stain were carried out on one of them. The feasibility of the thermal treatments in the field was verified with a seventh species. The results confirm that a 6 h treatment at 55 °C is sufficient to kill the lichens if they are kept fully hydrated. At 40 °C the organisms are damaged: in this case biocides at concentrations 10× lower than in normal applications can profitably be used. The new protocol is simple, the field equipment cheap, and the negative effects associated with standard biocide treatments are absent.     

Evaluation of peracetic acid sanitizers efficiency against spores isolated from spoiled cans in suspension and on stainless steel surfaces

The aim of this study was to determine the inactivation effect of industrial formulations of peracetic acid biocides on bacterial spores adhering to stainless steel surfaces. A standardized protocol was used to validate biocide activity against spores in suspension. To validate sporicidal activity under practical conditions, we developed an additional protocol to simulate industrial sanitization of stainless steel surfaces with a foam sanitizer. Spores of three spore-forming bacteria, Clostridium sporogenes PA3679, Geobacillus stearothermophilus, and Moorella thermoacetica/thermoautotrophica, were sprayed onto stainless steel as bioaerosols. Sporicidal activity was high against the C. sporogenes spore suspension, with more than 5 log CFU ml(-1) destroyed at all liquid biocide contact times. Sporicidal activity also was high against G. stearothermophilus and M. thermoacetica/thermoautotrophica spores after 30 min of contact, but we found no population reduction at the 5-min contact time for the highest sporicide concentration tested. The foam biocide effectively inactivated C. sporogenes spores adhered to stainless steel but had a reduced decontamination effect on other species. For G. stearothermophilus spores, sanitization with the foam sporicide was more efficient on horizontal steel than on vertical steel, but foam sanitization was ineffective against M. thermoacetica/thermoautotrophica whatever the position. These results highlight that decontamination efficiency may differ depending on whether spores are suspended in an aqueous solution or adhered to a stainless steel surface. Biocide efficiency must be validated using relevant protocols and bacteria representative of the microbiological challenges and issues affecting each food industry.     

Inactivation of Lactobacillus helveticus bacteriophages by thermal and chemical treatments.

The effect of several biocides and thermal treatments on the viability of four Lactobacillus helveticus phages was investigated. Times to achieve 99% inactivation of phages at 63 degrees C and 72 degrees C in three suspension media were calculated. The three suspension media were tris magnesium gelatin buffer (10 mM Tris-HCl, 10 mM MgSO4, and 0.1% wt/vol gelatin), reconstituted skim milk sterile reconstituted commercial nonfat dry skim milk, and Man Rogosa Sharpe broth. The thermal resistance depended on the phage considered, but a treatment of 5 min at 90 degrees C produced a total inactivation of high titer suspensions of all phages studied. The results obtained for the three tested media did not allow us to establish a clear difference among them, since some phages were more heat resistant in Man Rogosa Sharpe broth and others in tris magnesium gelatin buffer. From the investigation on biocides, we established that sodium hypochlorite at a concentration of 100 ppm was very effective in inactivating phages. The suitability of ethanol 75%, commonly used to disinfect utensils and laboratory equipment, was confirmed. Isopropanol turned out to be, in general, less effective than ethanol at the assayed concentrations. In contrast, peracetic acid (0.15%) was found to be an effective biocide for the complete inactivation of all phages studied after 5 min of exposure. The results allowed us to establish a basis for adopting the most effective thermal and chemical treatments for inactivating phages in dairy plant and laboratory environments.     

An inorganic biocide using a novel presentation of silver

A silver-based preservative system has been developed which overcomes difficulties often associated with metals, namely solubility, dispersion and toxicity. It comprises a silver chloride titanium dioxide composite which allows the controlled release of silver ions. Microbiological data has shown the composite to be active at concentrations less than 25 ppm against a wide range of micro-organisms, including Gram-negative non-fermentative bacteria, yeasts and moulds. The composite has satisfactory preservative activity against bacteria and fungi at application concentrations of between 50 ppm and 500 ppm, depending on the formulation. At high concentrations (>300 ppm) there may be some grey discoloration of the product, but the composite remains effective. To overcome this problem, a system of enhancing antimicrobial activity was devised, based on a sulphosuccinate salt dispersing agent. This allows lower concentrations of composite to be used whilst maintaining equivalent antimicrobial activity.     

Resistance of phages lytic to pathogenic Escherichia coli to sanitisers used by the food industry and in home settings

Phages are potentially useful as antimicrobial agents in food‐processing environments, provided they can remain active upon extended storage and in the presence of sanitisers. Survival of six phages lytic against enteropathogenic (EPEC) and shiga‐toxigenic (STEC) Escherichia coli strains was assessed upon storage at 4 °C, ?20 °C and ?70 °C in phosphate‐buffered‐saline (PBS) and Tris‐magnesium‐gelatine buffer (TMG) for up to 1 year. The phages were also exposed to ethanol, sodium hypochlorite, peracetic acid, quaternary ammonium chloride (biocide A), hydrogen peroxide/peracetic acid/peroctanoic acid (biocide B), p‐toluensulfonchloroamide (biocide D) and alkaline chloride foam (biocide C). Plaque‐forming units remained unchanged when the phages were stored at 4 °C in both buffers tested, but decreased by 3.5 and 5.7 log₁₀ PFU when stored in PBS at ?20 and ?70 °C, respectively. Moreover, TMG seems to be the most protective suspension medium with decreasing temperature because a 1?log₁₀ PFU reduction was observed at ?20 and ?70 °C. Incubation in 100% ethanol for 24 h reduced plaque counts only by 2.5 log₁₀ PFU. In 10 ppm of sodium hypochlorite and in biocide B (0.13%), the counts decreased by ~5 and ~6 log₁₀ PFU after 15 min. Finally, biocide A and D reduced counts by 4 and 2–4 log₁₀ PFU after 30 min and 8 h of incubation, respectively. Phages were completely inactivated only by peracetic acid and biocides C and E. Therefore, the phages evaluated might be potentially applied together with classical sanitisers such as ethanol and industrial biocides A, B and D, in disinfection programs against pathogenic STEC and EPEC strains.     

Inactivation of Lactobacillus delbrueckii bacteriophages by heat and biocides

The effect of several biocides and thermal treatments on the viability of four Lactobacillus delbrueckii phages was investigated. Time to achieve 99% inactivation of phages at 63 and 72 degrees C in three suspension media (Tris Magnesium Gelatin (TMG) buffer, Man Rogosa Sharpe (MRS) broth and reconstituted nonfat dry skim milk (RSM)) was calculated. Thermal resistance depended on the phage considered, but a marked heat-resistance was exhibited by one phage (Ib(3)) since its high titre suspensions were completely inactivated only after 45 min at 72 degrees C or 15 min at 90 degrees C. A clear protective effect of the milk was revealed when the three suspension media were compared. As regards to the effects of biocides on phages, only peracetic acid was found to be effective for inactivating high titre suspensions. Ethanol, even at a concentration of 100%, was not suitable to assure no surviving phage particles and isopropanol turned out to be less effective than ethanol. Sodium hypochlorite at 200-400 ppm inactivated the phages completely, except phage Ib(3), which was only destroyed after treatments with 1200 ppm. The diversity observed in the heat and biocide resistance of L. delbrueckii phages is useful to establish a basis for adopting the most effective thermal and chemical treatments for inactivating them in dairy plants and laboratory environments.     

Post-harvest kernel discoloration and fungi activity in long-grain hybrid,pureline and medium-grain rice cultivars as influenced by storage environment and antifungal treatment

Milled rice grade is diminished as the quantity of discolored kernels increases. This study evaluated the magnitude of kernel discoloration and relationship to fungal growth for long-grain hybrid (XL 753), long-grain pureline (Roy J.) and medium-grain (Titan) rice, exposed to storage moisture contents (MCs) of 16 and 21%, wet basis, temperatures of 20, 30, 40 °C, and durations up to 16 weeks. Impact of postharvest treatments with the antifungal agents, natamycin and sodium chloride, on rice discoloration and fungal growth was compared with untreated (control) samples. Subsamples were taken every four weeks to determine total fungal counts, discoloration, and water activity. Rice stored at 40 °C had six-to eleven-times more discoloration than at 20 °C. Increased storage duration significantly increased discoloration. Discoloration was not different between MC levels. Discoloration was significantly lower by sodium chloride treatment, while natamycin was not effective compared to control. Rice samples treated with sodium chloride also had significantly less water activity (mean a_w = 0.73) compared to samples treated with natamycin (mean = 0.84) and control samples (mean = 0.83). Hybrid rice had significantly less discoloration compared to pureline and medium-grain. Fungal counts were similar across all cultivars. Increasing temperature and duration decreased fungal count. An inverse relation between fungi and discoloration suggests microbe activities did not explain discoloration. The findings provide important information to growers and processors on conditions necessary to maintain rice quality.     

Efficacy of chitosan, carvacrol, and a hydrogen peroxide-based biocide against foodborne microorganisms in suspension and adhered to stainless steel

The ability of natural compounds to inactivate foodborne organisms adhered to surfaces was investigated with the ultimate aim of replacing synthetic biocides by more environmentally friendly, natural alternatives. The antimicrobial efficacy of 0.5, 1.0, and 2.0% chitosan and Spor-Klenz RTU (a commercial biocide based on hydrogen peroxide and peroxyacetic acid) and 0.5, 1.25, and 2.0 mM carvacrol was determined at 20 degrees C against Listeria monocytogenes, Salmonella enterica serovar Typhimurium, Staphylococcus aureus, and Saccharomyces cerevisiae adhered to stainless steel disks. Treatment with up to 2.0% chitosan reduced the viable cell count in the microbial films of the four test organisms by 2.4, 1.8, 2.3, and 0.9 log CFU/test surface (t.s.), respectively. By contrast, planktonic counts of the same organisms were reduced by 0.8 to 1.7 log CFU/ml at 2.0% chitosan. Treatment with 2 mM carvacrol reduced the viable counts of adhered listeriae, salmonellae, and yeasts by 2 to 3 log CFU/t.s. but S. aureus counts were reduced by only 0.9 log CFU/t.s. The efficacy of any single compound was species specific. In the case of microbial films prepared using listeriae and salmonellae, Spor-Klenz RTU was most biocidal, followed by carvacrol and then chitosan. However, dried films of S. aureus were most sensitive to chitosan and relatively resistant to carvacrol and Spor-Klenz RTU. By contrast, yeast films were most sensitive to carvacrol and least sensitive to chitosan. It was concluded that carvacrol and chitosan may have potential for use as natural biocides although optimization of conditions would be necessary.     

Inhibitory effect of seven Allium plants upon three Aspergillus species.

Antifungal activity and minimal fungicidal concentration (MFC) of extracts of garlic, bakeri garlic, Chinese leek, Chinese chive, scallion, onion bulb and shallot bulb against Aspergillus niger, A. flavus and A. fumigatus were examined. These Allium plants possessed antifungal activity, with garlic showing the lowest MFC. With the exception of scallion, the inhibitory effect of Allium plants against three Aspergillus species decreased with increasing incubation and heating temperature (P < 0.05). Acetic acid treatments of the extracts increased the inhibitory effect for all plants against three fungi (P < 0.05), and there was no significant difference in this effect among the three pH (2, 4, 6) treatments (P > 0.05) investigated. Acetic acid, at pH = 4, plus heat treatments of the extracts resulted in a greater inhibitory effect for all Allium plants against the three fungi than heat treatment alone (P < 0.05). Treatments of the extracts with NaCl, at concentrations of 0.2 and 0.4 M, did not affect the inhibitory effect of the plant extracts. The combination of acetic acid plus Allium plants was indicated to be an effective way to inhibit fungal growth.     

Quaternary ammonium compound stresses induce specific variations in fatty acid composition of Pseudomonas aeruginosa

The involvement of cell membrane fatty acids in resistance of Pseudomonas aeruginosa to Quaternary Ammonium Compounds (QACs) stresses was investigated. The strain was grown in a medium with increasing concentrations of different biocides: two QACs, and two non-QACs. In the presence of two QACs only, the strain was able to grow with increasing concentrations. During cellular adaptation to QACs, the resistance to the same biocide increased. A principal component analysis was performed with whole of fatty acid compositions which highlighted a specific variation for the cultures in presence of QACs. These modifications gave evidence of the outer membrane involvement in cellular response to the presence of QACs.     

Biofilm formation and biocides sensitivity of Pseudomonas marginalis isolated from a maple sap collection system

The susceptibility of planktonic and biofilm cells of Pseudomonas marginalis toward four commonly used biocides at different temperatures (15 and 30 degrees C) and biofilm growth times (24 and 48 h) was assessed. Using the MBEC biofilm device, biofilm production in maple sap was shown to be highly reproducible for each set of conditions tested. Biofilm formation was influenced by growth temperature and time. A temperature of 15 degrees C and incubation time of 24 h yielded fewer CFU per peg and showed fewer adhered cells and typical biofilm structures, based on scanning electron microscopy observations as compared with other conditions. Minimal biofilm eradication concentration values for P. marginalis were significantly greater (P. < 0.001) than were MBCs for planktonic cells and for every biocide tested, with the exception of minimal biofilm eradication concentration values for peracetic acid at 15 degrees C and 24 h. Sodium hypochlorite and peracetic acid sanitizers were able to eliminate P. marginalis biofilms at lower concentrations as compared with hydrogen peroxide- and quaternary ammonium-based sanitizers (P < 0.001). According to the results obtained, sodium hypochlorite and peracetic acid sanitizers would be more appropriate for maple sap collection system sanitation.     

Uptake of copper,nickel and zinc by crops growing in contaminated soils

A series of three replicated pot trials is reported in which various crops were grown in soils having enhanced concentrations of copper, nickel and zinc. Concentrations of these elements in the tops of plants harvested at their most sensitive stage were compared with ‘total’ and ‘extractable’ concentrations in soil and with concentrations in soil solutions. There was little difference between the relationships of ‘total’ and ‘extractable’ soil metal and concentrations in plant tissue. In general, the correlation between concentrations of metals in soils and plants was unpredictable. Plants differed in their efficiency of uptake of elements; lettuce assimilated more than the other crops tested (barley, rape and ryegrass). Similarly, soil concentrations of the elements required to achieve toxic thresholds in plant tops increased in the order lettuce, ryegrass, rape and barley. Measurements made with conventional extractants of copper, nickel and zinc in soils can be of value in predicting plant uptake and hence toxicity, only if appropriate calibration curves plotting extractable soil metal against plant uptake are at hand for the particular soils and plants under consideration. Mild extracts are more sensitive to the soil properties, especially pH value, which determine plant uptake and results with metals in soil solution were promising, especially for zinc. Nevertheless, soil analyses for copper, nickel and zinc are not always closely associated with their likely toxicity to crops.     

Effect of carbendazim and physicochemical factors on the growth and ochratoxin A production of Aspergillus carbonarius isolated from grapes

Carbendazim is a systemic fungicide that is commonly used on several crops (tobacco, fruit, vegetables, cereals, etc.). This fungicide is used to control fungal infections in vineyards. It is indicated against Botrytis cinerea, Uncinula necator, Plasmopara viticola and other fungi and can be used either alone or coupled with other fungicides. However, there is a lack of in-depth studies to evaluate its effectiveness against growth of Aspergillus carbonarius isolated from grapes and OTA production. A medium based on red grape juice was used in this study. Preliminary studies were performed at 0.98 a(w) and 25 degrees C using carbendazim concentrations over a wide range (1-2000 ng/ml medium) to control both growth of a strain of A. carbonarius isolated from grape and its ability to produce OTA. As the lag phase increased considerably at levels > 1000 ng/ml of medium, detailed studies were carried out in the range 50-450 ng/ml of medium at 0.98-0.94 a(w) and 20-28 degrees C. Statistical analysis (multifactor ANOVA) of the data revealed that the three factors assayed and the interactions a(w)-carbendazim concentration and a(w)-temperature had significant effects on lag phase duration. The highest lag-times were observed at 0.94 a(w,) 20 degrees C, and with 450 ng carbendazim/ml. The three factors also had significant effects of the growth rate and there was an interaction between a(w) and temperature. The growth rate of A. carbonarius in these cultures is favoured by high water availability and relatively high temperatures. However, addition of carbendazim at the assayed levels did not significantly influenced fungal growth rate. Accumulation of OTA was studied as a function of four factors (the three previously considered, and time). All factors had significant effects on the accumulation of OTA. There were also two significant interactions (a(w)-temperature and temperature-time). On the basis of the results obtained, carbendazim does not increase the lag phase of A. carbonarius except at relatively low a(w) and temperatures. It does not substantially decrease fungal growth rate once growth is apparent but it appears to cause an increase in OTA accumulation in the medium at the doses assayed. Carbendazim, which is widely used against fungal infections in grape, can positively influence OTA production by A. carbonarius in field, which can increase OTA content in grape juices and wines.     

Phenolic compounds and plant extracts protect paprika against UV-induced discoloration

The efficiency of phenolic antioxidants in protecting paprika carotenoids against discoloration was studied in a model gel system that was exposed to radiation of 274 nm at 20 °C. The fading of the colour of the gel was followed over a 6‐h period. Fifty percent of the colour faded from the control after 70 min of radiation. The ability of each antioxidant to prevent discoloration was assessed by using several different concentrations. Antioxidant compounds can be ranked according to their protective effect on colour: BHT > α‐tocopherol > epigallocatechin gallate > quercetin > rosmarinic acid ≥ caffeic acid and ferulic acid > coumaric acid > catechin. The ability of rosemary and tea extracts to prevent discoloration was comparable to pure compounds and this could be explained by their polyphenol content. Mixtures of α‐tocopherol with rosemary extract revealed strong synergistic effects.     

Potential of hydroxytyrosol-rich composition from olive mill wastewater as a natural disinfectant and its effect on seeds vigour response

Hydroxytyrosol-rich olive mill wastewater (HROMW) and hydroxytyrosol-rich composition (HRC) were prepared from olive mill wastewater using hydrolysis and post-hydrolysis purification processes. The HROMW and HRC showed powerful bactericidal and fungicidal activities against phytopathogens, and their minimal inhibition concentrations for fungi and bacteria were 7.18–57.4 mg l⁻¹ and 7.18–14.4 mg l⁻¹, respectively. After 5 min of contact time, the disinfectant properties of the HROMW and HRC added at concentrations of 1.25% (dw/v) allowed for a reduction in bacterial viability by greater than 5 log units. However, a higher concentration of 1.5–3% (dw/v) or a longer contact time of 30 min were needed to achieve values for fungal viability reduction that were higher than the 4 log units recommended by EN 1275 [EN 1275 (1997a). Chemical disinfectants and antiseptics. Basic fungicidial activity. Test method and requirements (phase 1)]. HROMW and HRC were less effective with the most ubiquitous fungi, Fusarium spp., which needed 10% dw/v. The addition of HRC at 10% (dw/v) showed that the composition was a potent exogenous enhancer of growth that stimulated the seedling vigour of tomato and muskmelon, according to the conventional agronomic parameters for seed vigour. Compared to the control, the germination percentage, shoot weight, shoot height, and root length were all significantly enhanced in the HRC-treated seed plants. HRC was found to have effective disinfectant properties against seed-borne diseases. In treated seeds, the composition had significant effects on the control of damping-off disease groups at the pre-germination stage. HRC was also equally effective in the control of root rot diseases caused by Fusarium sambucinum and Alternaria solani as well as of wilts and even bacterial seed-borne pathogens. HROMW was also found to be as effective as HRC in terms of its efficacy against the three seed-borne diseases mentioned above.     

不同氨基酸螯合锌对小鼠抗氧化能力的影响

研究了三种形式锌源对小鼠抗氧化能力的影响。选用断奶小鼠(KM种)81只,随机分为三组(硫酸锌组、蛋氨酸锌组和亮氨酸锌组)。实验第一周,饲喂缺锌日粮。实验第8d起,三组饲喂加锌日粮。三种锌源均能提高肝脏、血清锌含量。补锌能使血清MDA含量明显下降,不同锌源间差异不显著(p>0.05)。补锌7d后,三种锌源提高了小鼠总抗氧化能力、总SOD、铜锌超氧化物歧化酶活性、谷胱甘肽含量,蛋氨锌组和亮氨酸锌组优于硫酸锌组酸(p<0.05)。亮氨酸锌,蛋氨酸锌的抗氧化能力优于硫酸锌,氨基酸螯合锌组之间抗氧化能力的差异不大。     

Combined effects of NaCl, NaOH, and biocides (monolaurin or lauric acid) on inactivation of Listeria monocytogenes and Pseudomonas spp

This study highlighted combinations of chemical stresses that could decrease or eliminate Listeria monocytogenes and Pseudomonas spp. surviving in food processing plants. Strains of L. monocytogenes, Pseudomonas fragi, and Pseudomonas fluorescens isolated from processing environments (meat and milk) were grown at 20 degrees C up to the early stationary phase. The strains were then subjected to 30 min of physicochemical treatments. These treatments included individual or combined acid (acetic acid), alkaline (NaOH), osmotic (NaCl), and biocides (fatty acids) challenges. Survival of the strains was studied after individual or combined acid (acetic acid), alkaline (NaOH), osmotic (NaCl), and biocides (monolaurin, lauric acid) challenges. Individual pH shocks had lower efficiencies than those used in combinations with other parameters. The treatment pH 5.4 followed by pH 10.5 had a low efficiency against L. monocytogenes. The opposite combination, pH 10.5 followed by pH 5.4, led to a 3-log reduction of the L. monocytogenes population. Pseudomonas spp. strains were much more sensitive than L. monocytogenes, and population reductions of 5 and 8 log (total destruction), respectively, were observed after the same treatments. As for L. monocytogenes, the combination pH 10.5 followed by pH 5.4 is more deleterious than the opposite. Whatever the bacterial species, the most efficient treatments were combinations of alkaline, osmotic, and biocide shocks. For instance, the combination pH 10.5 and 10% NaCl plus biocides showed reductions of 5 to 8 log for both bacteria. The origins of the observed lethal effects are discussed.