Acta Sci. Pol., Technol. Aliment. 12(4) 2013, 385-393 IMPEDIMETRIC
Total Page:16
File Type:pdf, Size:1020Kb
M PO RU LO IA N T O N R E U Acta Sci. Pol., Technol. Aliment. 12(4) 2013, 385-393 I M C S ACTA pISSN 1644-0730 eISSN 1889-9594 www.food.actapol.net/ IMPEDIMETRIC TEST FOR RAPID DETERMINATION OF PERFORMIC ACID (PFA) BIOCIDAL ACTIVITY TOWARD ECHERICHIA COLI Małgorzata Lasik1, Renata Dobrucka2, Piotr Konieczny1 1Faculty of Food Sciences and Nutrition, Poznań University of Life Sciences Wojska Polskiego 28, 60-637 Poznań, Poland 2Department of Industrial Products Quality and Ecology, Poznań University of Economics Niepodległości 10, 61-875 Poznań, Poland ABSTRACT Background. Performic acid has recently become available on a commercial scale for potential use in waste- water disinfection and can become an innovative biocide for various purposes in food processing. The aim of our study was: 1) to investigate the antimicrobial resistance of performic acid as high active and non toxic chemical disinfectant against Escherichi coli (hygiene indicator test microorganism used in industrial micro- biology) and 2) to evaluate the electrical impedance measurement method usefulness for fast and high precise test of antibacterial activity. Material and methods. Four types of antimicrobial disinfectants (commercial 35% hydrogen peroxide, 1% performic acid, 35% hydrogen peroxide and 15% formic acid) were tested against Escherichia coli as hy- giene indicator test microorganism. By evaluating the biocidal activity of selected disinfectants two methods were compared: electrical impedance measurement and classical serial dilution method with turbidity effect. Results. It was stated that the performic acid expressed the highest antibacterial activity in comparison to other tested peroxide disinfectants: commercial 35% hydrogen peroxide solution and components required for performic acid production: 35% hydrogen peroxide solution with stabilizers and 15% formic acid solution with stabilizers). It was demonstrated that the proposed alternative microbiology method of electrical imped- ance measurement facilitates a rapidly and more precise analyses of the intensity of disinfectants inhibition effect. Conclusions. It can be postulated that both, the performic acid disinfectants as well as the impedimetric method can be a good advantage in the industrial microbiology. Key words: performic acid, impedance measurement, disinfection, biocides INTRODUCTION Performic acid (CAS registry number and name: in different aspects. It was assumed that those com- 107-32-4, methaneperoxoic acid) is a mixture of two pounds have a stronger effect on airborne microbes that components: solution of hydrogen peroxide (35% so- other peroxides and oxidative substances. Their disin- lution of H2O2) and formic acid (concentrations from fective activity is associated with release of active oxy- 10 to 20%) with stabilizing substances, mixed in the gen [Baldry et al. 1991, Lefevre et al. 1992]. Probably ratio 1:1. So far research concerning the infl uence of trioxide binding, as well as thiol groups which can be peroxide acids on microorganisms has been carried out found in enzymes, structural proteins and other building [email protected] © Copyright by Wydawnictwo Uniwersytetu Przyrodniczego w Poznaniu Lasik M., Dobrucka R., Konieczny P., 2013. Impedimetric test for rapid determination of performic acid (PFA) biocidal activity toward Echerichia coli. Acta Sci. Pol., Technol. Aliment. 12(4), 385-393. compounds are oxidized and destroyed. The peroxide and Plockova [1995] investigated inhibiting properties acids activate for example catalase – enzyme known of honey and its infl uence on the metabolic activities for detox of free radicals [Lefevre et al. 1992, Viss- of lactic acid bacteria, while Paquet et al. [2000] ap- ers et al. 2009]. Double bonds are also oxidized. Acid, plied the measurement of electrical impedance to as- which penetrates into the inferior of a cell impedes the sess the activity of starter cultures in the process of basic metabolic traces and the active transport through cheese production. Whereas Lasik et al. [2002] and membranes. The compound has also the ability to affect Nowak et al. [2005] used the impedance technique to DNA [Baldry et al. 1991, Lefevre et al. 1992]. evaluate the inhibition effect of high loaded food in- Performic acid (PFA) is also one of the peroxide dustry wastewater on single and mixed bacteria popu- compounds, and can be treated as an alternative to oth- lations growth kinetic. er bactericidal substances, e.g. for the peracetic acid In the present study two main research aims were [Gehr et al. 2009]. Performic acid and peracetic acid stated. The fi rst aim was to investigate the antimicro- are rather similar disinfectant compounds. The effi - bial resistance of performic acid as high active and ciency of both these compounds base on active oxygen non toxic chemical disinfectant against Escherichi operation [Rutala and Weber 2001]. Performic acid has coli. This bacterium was chosen in our experiment recently become available on a commercial scale for as hygiene indicator test microorganism used in in- potential use in wastewater disinfection [Gehr et al. dustrial microbiology. And the other aim of the study 2009, Vissers et al. 2009]. Furthermore a wide poten- was to evaluate the possibility of electrical impedance tial of PFA as an innovative biocide for various purpos- measurement method used for fast and high precision es in food processing related applications was already test of antibacterial activity. The proposed alternative discussed by Dobrucka et al. [2009]. The substance in- microbiology method was compared to classical serial activates viruses, bacteria and bacterial spores, bacilli dilution method. The parameters of minimal ihhibitory and microscopic fungus and exhibit high antibacte- concentration and minimal bactericidal concentration rial activity also in low temperatures [McWilliam and (as results of classical method) were confronted with Stewart 2002, Heinonen-Tanski and Miettinen 2010]. impedance changes profi le and impedance detection In the present study the antibacterial activity of time (as result of impedimetric method). performic acid was evaluated by employing the elec- trical impedance measurement method – an alternative MATERIAL AND METHODS for the classical serial dilution method. The principle of the impedimetric technique is to measure electri- Disinfectant agents cal impedance changes caused by the microbial activ- Four types of antimicrobial disinfectant agents were ity during growth in the culture medium. Molecules tested: A. commercial 35% hydrogen peroxide solu- of high molecular weight present in the medium (e.g. tion, B. 1% performic acid solution and components proteins, carbohydrates and fats) are broken down by required for its production: C. 35% hydrogen peroxide viable microorganisms to ionized metabolic products. solution with stabilizers and D. 15% formic acid solu- Smaller, charged molecules result in an increase in tion with stabilizers. 1% performic acid solution was electric current conductivity and consequently lead to prepared by mixing 35% hydrogen peroxide solution a decrease in the electrical impedance of the culture (C) and 15% formic acid solution (D), in the ratio 1:1. environment [Gomez et al. 2002, Noble 1999, Yunus et al. 2002, Nowak et al. 2005]. In general, this method Microorganisms is used for rapid detection or quality and quantity anal- The strain of Escherichia coli PCM 318 O2:K1(L): yses of microorganisms, especially in the quality con- H4 from a strain collection of the Institute of Immu- trol of food, pharmaceuticals and cosmetics [Wawerla nology and Experimental Therapy in Wrocław, Poland et al. 1999, Moldenhauer 2003]. However, attempts was used as the test microorganism in the performed have been made to employ impedance measurements investigations. The test bacteria were cultured and test- to characterize the stimulating or inhibiting effect ed using a nutritional broth (BTL, Poland) as growth on the metabolic activities of microorganisms. Curda medium at 37°C for 48 h. 386 www.food.actapol.net/ Lasik M., Dobrucka R., Konieczny P., 2013. Impedimetric test for rapid determination of performic acid (PFA) biocidal activity toward Echerichia coli. Acta Sci. Pol., Technol. Aliment. 12(4), 385-393. Determination of minimal inhibitory bacteria during growth in the media contained tested concentration (MIC) disinfectants agents were measured in the Automatic The minimal inhibitory concentration (MIC) is Analyzer of Microorganisms Growth, BacTrac 4100 assumed to be the lowest concentration, at which no (Sy-Lab, Austria). Special 10-ml measuring test tubes microbial growth was observed [Wiegant et al. 2008, (Sy-Lab) equipped with 4 electrodes were used in the Mongalo et al. 2013, Sun at al. 2012]. Two-fold serial experiment. The test tubes were fi lled with 9 ml of dilutions of the four tested disinfectant agents were growth medium containing the antimicrobial agent, prepared for their antibacterial resistance evaluation. and then inoculated with 1 ml inoculum of the tested The serial dilutions were prepared in test tubes con- bacteria. The measuring test tubes were incubated at taining the bacteria growth medium and tested disin- 37°C in a thermostat of the Automatic Analyzer of fectant agents in a certain concentration. The lowest Microorganisms Growth. Changes in the electrical im- concentration of the tested agents that inhibited the pedance were calculated in relation to the initial value visible growth of E. coli after 48 h incubation was de- according to the following