Journal of Food Processing and Preservation ISSN 1745-4549

ANTIMICROBIAL ACTIVITY OF LACTIC ACID AGAINST

PATHOGEN AND SPOILAGE MICROORGANISMS
SLOBODANKA STANOJEVIĆ-NIKOLIĆ1, GORDANA DIMIĆ1, LJILJANA MOJOVIĆ2, JELENA PEJIN1,
ALEKSANDRA DJUKIĆ-VUKOVIĆ2 and SUNČICA KOCIĆ-TANACKOV1,3
1
Faculty of Technology, University of Novi Sad, Bulevar cara Lazara 1, Novi Sad 21 000, Serbia
2
Faculty of Technology and Metallurgy, University of Belgrade, Belgrade, Serbia

3
Corresponding author. ABSTRACT
TEL: +381 214853699;
FAX: +381 21450413; Preliminary examination of the antimicrobial activity of lactic acid against nine
EMAIL: [email protected] bacteria (Escherichia coli, Proteus mirabilis, Salmonella enteritidis, Pseudomonas
aeruginosa, Staphylococcus aureus, Enterococcus faecalis, Listeria monocytogenes,
Received for Publication July 21, 2015
Bacillus cereus and Bacillus megaterium) and three yeasts (Rhodotorula sp., Saccha-
Accepted for Publication October 20, 2015
romyces cerevisiae and Candida albicans) was performed using disc diffusion and
doi:10.1111/jfpp.12679 broth microdilution method. At a concentration of 321 mg/mL, inhibition zones
ranged from 24.0 mm (Es. coli) to 38.3 mm (En. faecalis) for the tested bacteria.
The inhibition zones of the yeasts ranged from 11.3 mm (Sac. cerevisiae) to
14.0 mm (Rhodotorula sp.).
Lactic acid minimal inhibitory concentration for bacteria was ≥1.25 mg/mL, while
minimal biocide concentration was ≥2.50 mg/mL. Minimal inhibitory concentra-
tion for yeasts was ≥12.50 mg/mL, while minimal fungicidal concentration was
≥25.00 mg/mL. The obtained results showed that lactic acid could be used as an
efficient natural antimicrobial agent improving the safety of all-natural foods.

PRACTICAL APPLICATIONS
Consumer perception that synthetic food additives may be associated with poten-
tial toxicological problems has recently generated interest for the use of naturally
derived compounds in the food industry. The use of lactic acid is considered as a
good alternative and may be more acceptable to consumers than synthetic food
additives because of its natural origin, potential antimicrobial activity, as well as
preservative, antioxidant, flavoring and acidifying properties as well as low cost.
For ensuring food safety, combination of lactic acid with other natural antimicro-
bial agents or other preservation methods should be considered. Therefore, there
is a potential for lactic acid usage in the development of eco-friendly technology
which ensures food safety.

agents into food products has been widely applied in the

INTRODUCTION
food industry (Brul and Coote 1999). The need for natural
Pathogenic and food spoilage microorganisms have been alternative is due to consumers’ preference for fewer chemi-
considered as the primary causes of foodborne diseases and cals and more natural foods. Lactic acid has been consid-
food quality deterioration, which results in many deaths ered as a potential alternative. The U.S. Food and Drug
and significant economic loss each year (Over et al. 2009). Administration classified lactic acid as “generally recognized
Consequently, there is a considerable interest in ways to as safe” (GRAS). Lactic acid is a natural substance found in
stop this upward trend and reduce the incidence of food various fruits and fermented products and exhibits antimi-
poisoning. crobial activity against foodborne pathogens (Beuchat and
In order to ensure the food safety and to extend the shelf Colden 1989). Moreover, numerous applications for decon-
life of food products, addition of chemical preservative tamination of meat, fruits and vegetables by lactic acid have

Journal of Food Processing and Preservation •• (2015) ••–•• © 2015 Wiley Periodicals, Inc. 1
ANTIMICROBIAL ACTIVITY OF LACTIC ACID S. STANOJEVIĆ-NIKOLIĆ ET AL.

been previously described (Dickson and Anderson 1992; Staphylococcus aureus (ATCC 1163), Enterococcus faecalis
Greer and Dilts 1995; Calicioglu et al. 2002; Gill and Badoni (ATCC 29212), Listeria monocytogenes (ATCC 19115), Bacil-
2004; Vasantha Rupasinghe et al. 2006; Alvarado-Casillas lus cereus (isolated from cheese), and B. megaterium (iso-
et al. 2007; Park et al. 2011; In et al. 2013). Besides its anti- lated from cheese) as well as the yeasts Rhodotorula sp.
microbial activity, lactic acid acts as a flavor enhancer and (isolated from air), Saccharomyces cerevisiae (ATCC 2601)
an antioxidant, preventing lipid oxidation by decreasing the and Candida albicans (ATCC 10231). Cultures were main-
pro-oxidative effect of NaCl (Paelinck and Szczepaniak tained on nutrient agar (NA) (bacteria) (Merck, Darmstadt,
2005). Germany) and on Sabouraud Dextrose agar (SDA) (yeasts)
Lactic acid is lethal to microorganisms via undissociated (Merck) at 4C as a part of the collection of the Laboratory
molecules that flow through the cell membranes and ionize for Food Microbiology at the Faculty of Technology, Univer-
inside. The acidic pH inside the cell causes deformation and sity of Novi Sad, Serbia.
damage to enzymatic activities, proteins and DNA struc- Bacterial strains were cultured on an NA for 24 h at 37C
ture, thereby damaging the extracellular membrane (or 30C for B. cereus and B. megaterium), while yeasts were
(Mani-Lopez et al. 2011). In another mechanism, changes in grown on SDA for 48 h at 25C. The suspensions were pre-
the permeability of the cell membrane hinder substrate pared in a sterile saline (0.85%; 10 mL) and adjusted (with
transport, while changes in the pH inside the cell suppress sterile saline) to give a final concentration of 106 cfu/mL
NADH oxidation; this affects the electron transport system using McFarland Standard (bioMérieux SA, Marcy I’Etoile,
and leads to the death of the microorganism (Kong et al. France) and McFarland densitometer (Biosan SIA, Riga,
2001). Latvia). Microorganism suspension concentration was also
Many studies showed the antimicrobial activity of lactic determined using agar plate method (using NA medium for
acid bacteria supernatant in which the predominant antimi- bacteria and SDA medium for yeasts).
crobial agent is lactic acid (Magnusson and Schnürer 2001;
Aslim et al. 2005; Nardi et al. 2005; Zdolec et al. 2007; Rouse
et al. 2008; Polak-Berecka et al. 2009; Wongsuttichote and Disc Diffusion Assay
Nitisinprasert 2009; Voulgari et al. 2010; Al Askari et al.
Preliminary examination of the lactic acid antimicrobial
2012; Köhler et al. 2012; Sungsri et al. 2012). However, there
activity was performed by a disc diffusion method (Leboffe
are only a few studies reporting the antimicrobial activities
and Pierce 2005). A sterile swab was immersed in the
of lactic acid against Enterococcus faecalis, Proteus mirabilis,
culture suspension (106 cfu/mL) and used to inoculate the
Bacillus megaterium, Candida albicans, Rhodotorula spp. and
entire surface of a Müller–Hinton agar (MHA) (Merck) (for
other toxic and spoilage microorganisms (Lind et al. 2005;
bacteria) and SDA (Merck) (for yeasts). Inoculated plates
Broberg et al. 2007; Pundir and Jain 2011).
were left for 15 min at a room temperature to allow good
Therefore, the aim of this study was to determine the in
absorption. Three or four 6-mm sterile filter paper discs
vitro antimicrobial activity of lactic acid against growth of
(Himedia, Mumbai, India) were placed equidistant apart on
nine bacteria and three yeasts that are frequently encoun-
the inoculated agar surface of the plates and certain
tered in food.
amounts of lactic acid (15.0, 20.0 and 25.0 μL of lactic acid
solution for concentrations of 102, 209 and 321 mg/mL, and
1.5, 2.5, 5.0 and 10.0 μL of pure lactic acid) were immedi-
MATERIALS AND METHODS
ately added. Sterile filter paper disc placed on the center of
the inoculated agar surface of the plates in which sterile
Preparation of Lactic Acid Solutions
water was added was used as a control.
L-(+)-lactic acid (Sigma-Aldrich Co. LLC, Taufkirchen, After 24 h of incubation at 37C (or 30C) for bacteria and
Germany) solutions were prepared with sterile distilled 48 h at 25C for yeasts, the inhibition zones were measured
water. Using acetate membrane filters (Sartorius SA, in millimeters.
Göttingen, Germany), pore size 0.2 μm, the solutions were
sterilized and stored at 4C up to 30 days.
Broth Microdilution Assay
Minimal inhibitory concentration (MIC), minimal bacteri-
Test Microorganisms, Culture Conditions and
cidal concentration (MBC) and minimal fungicidal concen-
Preparation of Suspension
tration (MFC) were determined according to the modified
Bacteria used in this study were Escherichia coli (ATCC method of Marcello et al. (2003) and Barbour et al. (2004).
10536), Proteus mirabilis (ATCC 12453), Salmonella enteriti- Müller–Hinton broth (MHB) (Merck) for bacteria and
dis (ATCC 13076), Pseudomonas aeruginosa (ATCC 10145), Sabouraud Dextrose broth (Merck) for yeasts (100 μL) were

2 Journal of Food Processing and Preservation •• (2015) ••–•• © 2015 Wiley Periodicals, Inc.
S. STANOJEVIĆ-NIKOLIĆ ET AL. ANTIMICROBIAL ACTIVITY OF LACTIC ACID

pipetted into each well of a sterile 96-well microtiter plate

RESULTS AND DISCUSSION
(Greiner Bio-One GmbH, Kremsmünster, Austria). After
that, in the first column of wells in each microtiter plate
Lactic Acid Antimicrobial Activity by Disc
100 μL of the initial concentration of lactic acid (5, 10, 20,
Diffusion Method
50, 75, 102 mg/mL for bacteria and 20, 50, 75, 102, 209,
321 mg/mL for yeasts) was transferred. After mixing by The results of the lactic acid antimicrobial activity obtained
pipetting, 100 μL of the mixture was transferred to the next by the disc diffusion method are given in Table 1. The
column of wells in a process of 1 : 1 serial dilution until the results showed that lactic acid has a potential to inhibit the
column number 12 (ranging from 0.15 to 6.37 mg/mL for growth of all tested bacteria and yeasts. Test microorgan-
bacteria and from 0.62 to 20.06 mg/mL for yeasts). Each isms showed different susceptibility.
well was then inoculated with 10 μL of the tested microor- The results of the antimicrobial activity of the different
ganism suspension (106 cfu/mL). lactic acid solution concentrations (102, 209 and 321 mg/
Three controls were used: first control contained appro- mL) on the growth of the test microorganisms obtained by
priate broth and lactic acid, second control contained broth disc diffusion method are given in Table 2. The inhibition
and suspension of tested microorganism, while third area increased with the increase in lactic acid concentration.
control contained only broth. Incubation of microtiter In general, lactic acid showed the strongest inhibitory
plates was performed at the appropriate temperature for activity against bacteria. Lactic acid was more effective
24 h (for bacteria) or 48 h (for yeasts). After incubation, against gram-positive bacteria than gram-negative bacteria.
wells that contained clear broth suspension were used for Among gram-positive bacteria, the most susceptible was En.
inoculation of predried MHA and SDA plates. faecalis, while Ps. aeruginosa was the most susceptible gram-
The MIC was determined as the lowest lactic acid con- negative bacteria. Similar susceptibility was obtained for
centration that inhibited the growth of the well (clear broth Es. coli, Sal. enteritidis and Sta. aureus.
suspension), but still showed slightly visible growth on the Some studies showed the antimicrobial effect of lactic
plate. MBC and MFC were determined at the lowest lactic acid against Es. coli, L. monocytogenes, Salmonella Gaminara
acid concentration that inhibited the growth of the well and (Eswaranandam et al. 2004), Sal. enteritidis (Anang et al.
showed no visible growth on the plate. 2007), Sta. aureus (Ho et al. 2010; Sudershan et al. 2011),
also against resistant clinical isolates of Ps. aeruginosa
(Jamalifar et al. 2011) and spore-forming bacteria (Jamilal
Statistical Analysis
et al. 2008; Tharmaray and Shah 2009; Ho et al. 2010).
The tests were performed in three replications. MS Excel Moreover, Ibrahim et al. (2008) found that lactic acid even
(Microsoft Corporation, Redmond, WA) was used to calcu- at low concentration (2 mg/mL) inhibits the growth of Sal-
late the means and standard deviations. monella spp. and Es. coli O157:H7. Similarly, Yesillik et al.

TABLE 1. PURE LACTIC ACID ANTIMICROBIAL

Pure lactic acid
ACTIVITY ON TESTED MICROORGANISMS
OBTAINED BY DISC DIFFUSION METHOD Amount (μL/disc) 1.5 2.5 5.0 10.0
Gram-negative bacteria
Escherichia coli 14 ± 1.11* 18 ± 0.00 24 ± 0.44 32 ± 0.66
Salmonella enteritidis 10 ± 0.53 16 ± 0.98 16 ± 0.98 26 ± 0.00
Proteus mirabilis 8 ± 0.89 11 ± 1.70 11 ± 1.70 20 ± 0.00
Pseudomonas aeruginosa 17 ± 0.55 35 ± 0.76 40 ± 0.87 50 ± 0.75
Gram-positive bacteria
Staphylococcus aureus 19 ± 0.00 20 ± 0.30 24 ± 0.00 30 ± 0.45
Enterococcus faecalis 18 ± 0.34 33 ± 0.88 33 ± 1.32 38 ± 0.32
Listeria monocytogenes 32 ± 0.12 37 ± 1.21 38 ± 0.45 40 ± 0.54
Bacillus cereus 20 ± 0/44 21 ± 0.00 33 ± 0.22 34 ± 0.12
Bacillus megaterium – 10 ± 0.00 16 ± 0.40 33 ± 0.22
Yeasts
Rhodotorula sp. – – 7 ± 0.12 12 ± 0.00
Candida albicans – – 7 ± 0.23 9 ± 0.34
Saccharomyces cerevisiae ci ci ci ci

* Standard deviation.
ci, complete inhibition; –, not sensitive.

Journal of Food Processing and Preservation •• (2015) ••–•• © 2015 Wiley Periodicals, Inc. 3
 ANTIMICROBIAL ACTIVITY OF LACTIC ACID S. STANOJEVIĆ-NIKOLIĆ ET AL.

(2011) showed that lactic acid at a concentration of

24.0 ± 0.00
27.6 ± 1.15
28.3 ± 0.57

25.0 ± 1.00
28.3 ± 0.57

38.3 ± 1.53
36.0 ± 0.00

14.0 ± 0.00
12.6 ± 0.57
11.3 ± 0.57
29.6 ± 1.15
30.0 ± 0.00
9 mg/mL inhibits the growth of Salmonella typhimurium

25.0
(inhibition zone of 22.6 mm) and Ps. aeruginosa (inhibition
zone of 22.5 mm), while against Es. coli and Sta. aureus
lactic acid was ineffective.
23.6 ± 1.15
23.0 ± 0.00
22.6 ± 1.15

24.0 ± 0.00

13.0 ± 1.00
10.3 ± 0.57
10.6 ± 1.15
24.3 ± 0.57

36.0 ± 1.00
31.0 ± 1.00
26.6 ± 0.57
28.3 ± 0.57
Several authors suggested that lactic acid is more efficient
antibacterial agent than acetic, citric and propionic acid
20.0

(Bjornsdottir et al. 2006; Pundir and Jain 2011; Daskalov

2012). Other studies indicate higher efficiency of sorbic and
benzoic acid than lactic acid (Yuk et al. 2007; Ho et al.
22.6 ± 1.15
21.6 ± 1.15
18.6 ± 1.15

20.6 ± 1.15
23.0 ± 1.00

30.3 ± 0.57
25.6 ± 0.57

11.0 ± 1.00
23.0 ± 0.00
21.3 ± 1.15
2010).
The obtained results showed that lactic acid is more effec-
15.0

0
0
321

tive against gram-positive bacteria than gram-negative bac-

teria, which is in agreement with the results obtained by
other authors (Houtsma et al. 1993; Yuk et al. 2007;
23.6 ± 1.15
24.3 ± 0.57
24.6 ± 1.15

24.0 ± 1.00

11.6 ± 1.15
10.0 ± 0.00
10.0 ± 0.00
24.6 ± 1.15

28.0 ± 1.00
20.3 ± 0.57
23.3 ± 1.15
29.3 ± 0.57

Tharmaray and Shah 2009). One reason for this is the dif-
ferent structural and chemical compositions of the cells’
TABLE 2. LACTIC ACID ANTIMICROBIAL ACTIVITY AGAINST TESTED BACTERIA AND YEASTS OBTAINED BY DISC DIFFUSION METHOD

25.0

outer layers (Nikaido and Vaara 1985) and acid tolerance of

each microorganism. In this sense, Nikaido (1996, 2003) has
reported that the resistance mechanisms in gram-negative
22.3 ± 0.57
21.6 ± 1.00
19.0 ± 0.00

21.6 ± 1.53

10.0 ± 0.00
23.6 ± 0.57

26.0 ± 0.00
17.6 ± 1.15
22.0 ± 1.00

7.6 ± 1.15
26.6 ± 0.57

bacteria are more complicated than those present in gram-

positive bacteria as the former is surrounded by the outer
20.0

membranes or a membrane that has hydrophobic nature,

which may block the entry of hydrophilic molecules of low
molecular mass such as monosaccharides, amino acids,
19.6 ± 1.73
19.0 ± 0.00
16.6 ± 1.15
18.6 ± 1.15

23.6 ± 0.57
18.6 ± 0.57

15.3 ± 0.57
20.6 ± 0.57
19.6 ± 1.15

8.0 ± 0.00

nucleosides and alkali or alkaline ions, which can only pass

through water-filled channels formed by transmembrane
15.0

0
0
209

proteins (porins) embedded into the lipid bilayer that

permits hydrophilic transport. On the one hand, the gram-
positive cell wall contains only a thick peptidoglycan layer
16.6 ± 1.15
15.6 ± 0.57
13.6 ± 1.15
17.0 ± 0.00

19.0 ± 0.00
16.0 ± 1.00

18.0 ± 0.00
20.3 ± 0.57
17.3 ± 0.57

10.3 ± 0.57
Diameter of inhibition zone (mm) (mean ± SD)

and a lipid bilayer. That fact would explain the greater sensi-
25.0

0
0

tivity of En. faecalis (gram-positive) to lactic acid than

gram-negative bacteria. On the other hand, Sta. aureus was
more resistant to lactic acid than En. faecalis. This is in
13.3 ± 0.57
14.0 ± 0.00
12.0 ± 0.00
16.6 ± 1.15

18.3 ± 0.57
15.3 ± 0.57

16.0 ± 1.00
18.0 ± 1.00
15.6 ± 0.57

8.3 ± 0.57

accordance with the results reported by other authors

(Miller and Kaspar 1994; Arnold and Kaspar 1995;
20.0

0
0

Benjamin and Datta 1995; Lin et al. 1996) who indicated

that acid resistance is a typical characteristic of the tested
microorganism.
11.3 ± 0.57
12.3 ± 0.57
10.3 ± 0.57
13.6 ± 1.15

15.6 ± 1.15
11.0 ± 1.73

13.6 ± 0.57
15.3 ± 0.57
12.0 ± 1.00

However, some authors found that effectiveness of the

organic acids can vary depending on its molecular weight.
15.0

0
0
0
102

Eswaranandam et al. (2004) indicated that undissociated

smaller molecules of lactic (90.08 Da) acid may enter into
the bacterial cells easily and change the internal pH of the
Lactic acid concentration (mg/mL)

microorganism, thus showing higher antimicrobial activity

than undissociated larger molecules of citric (192.13 Da)
Pseudomonas aeruginosa

Saccharomyces cerevisiae
Listeria monocytogenes
Staphylococcus aureus

and tartaric (150.09 Da) acids, which may not enter to the
Salmonella enteritidis
Gram-negative bacteria

Enterococcus faecalis

SD, standard deviation.

Gram-positive bacteria

Bacillus megaterium

cell interior effectively. That fact would explain the consid-

Candida albicans
Proteus mirabilis

Rhodotorula sp.
Escherichia coli

erable sensitivity of some gram-negative bacteria

Amount (μL/disc)

Bacillus cereus

(Ps. aeruginosa) to lactic acid.

In this study the most susceptible yeasts to lactic
Yeasts

acid was Rhodotorula sp., while similar susceptibility was

obtained for C. albicans and Sac. cerevisiae at all applied

4 Journal of Food Processing and Preservation •• (2015) ••–•• © 2015 Wiley Periodicals, Inc.
S. STANOJEVIĆ-NIKOLIĆ ET AL. ANTIMICROBIAL ACTIVITY OF LACTIC ACID

TABLE 3. MINIMAL INHIBITORY CONCENTRATION (MIC) AND from 1.25 mg/mL (Sal. enteritidis, Ps. aeruginosa and
MINIMAL BACTERICIDAL CONCENTRATION (MBC) OF LACTIC ACID L. monocytogenes) to 18.75 mg/mL (B. cereus), while for
FOR THE TESTED MICROORGANISM
yeasts MIC ranged from 12.50 mg/mL (Rhodotorula sp.) to
Microorganisms MIC (mg/mL) MBC (mg/mL) 37.50 mg/mL (Sac. cerevisiae). Lactic acid MBC ranged
Gram-negative bacteria from 2.50 to 37.50 mg/mL, while MFC ranged from 25.00
Pseudomonas aeruginosa 1.25 2.50 to 75.00 mg/mL (Table 3).
Salmonella enteritidis 1.25 2.50 Lactic acid showed high inhibitory effect against
Proteus mirabilis 2.50 5.00 L. monocytogenes, with a MIC of 1.25 mg/mL. The results of
Escherichia coli 2.50 5.00
MIC and MBC showed that two gram-negative bacteria
Gram-positive bacteria
(Sal. enteritidis and Ps. aeruginosa), with a MIC concentra-
Listeria monocytogenes 1.25 2.50
Staphylococcus aureus 2.50 5.00 tion of 1.25 mg/mL, were more sensitive to lactic acid than
Enterococcus feacalis 2.50 5.00 most gram-positive bacteria (Table 3).
Bacillus megaterium 12.50 25.00 Spore-forming bacteria were more resistant to the lactic
Bacillus cereus 18.75 37.50 acid effect than the other test bacteria. MIC and MBC for
Yeasts B. megaterium and B. cereus were significantly higher than
Rhodotorula sp. 12.50 25.00
those of the tested non-sporulating bacteria. Russell (1991)
Candida albicans 18.75 37.50
Saccharomyces cerevisiae 37.50 75.00
reported that organic acids added as food acidulants are
almost equally effective in inhibiting gram-positive and
gram-negative bacteria, while spore-forming bacteria are
usually more resistant to preservatives than are non-
concentrations. Some authors suggest that lactic acid is not sporulating bacteria (Gauld 1985; Russell 1990). The resis-
sufficiently effective as an antifungal agent (Woolford 1975; tance of spore-forming bacteria to preservatives is a
Wongsuttichote and Nitisinprasert 2009; Muck 2010), while property of the spore coats, although the cortex may be a
other studies indicate that lactic acid has the potential anti- contributing factor (Knott et al. 1990).
fungal activity (Lind et al. 2005; Broberg et al. 2007). Some Higher lactic concentrations, which are necessary for
yeasts can grow at lower pH values (Beales 2004) and at total microorganism inhibition, may cause changes in food
lower lactic acid concentrations (Narendranath et al. 2001), sensory properties. However, in many cases, a concentration
while others may use lactic acid as a nutrient (Moon 1983). sufficient to result in growth stasis may be all that is
Some yeasts (Sac. cerevisiae and C. albicans) use organic required to achieve a safe product, provided that the initial
acids by decarboxylation (Stratford et al. 2007). microorganism number is low (de Barros et al. 2012;
The variability in the sensitivity to weak organic acid pre- Tangwatcharin and Khopaibool 2012). Also, the concentra-
servatives, among the yeast species, may be related to their tion required for the growth stasis can be considerably
capacity to change the cell metabolism in response to acid lower than that necessary for the total inhibition.
stress conditions and physiological differences such as cell Inhibitory activity of the lactic acid against three strains
permeability, enzyme structure, and induction or suppres- of foodborne pathogenic bacteria was investigated by Qiao
sion of genes that effect susceptibility to weak acids (Warth et al. (2008). In comparison with our study, MIC of lactic
1991). Some yeasts possess enzymes able to degrade acids acid for Sta. aureus, B. cereus, and Es. coli ranged from
and reduce diffusion coefficient of preservatives across the 0.9 mg/mL (Es. coli) to 3.6 mg/mL (B. cereus), while MBC
membrane, resulting in a reduced ability of weak acids to ranged from 1.8 to 7.2 mg/mL. Hsiao and Siebert (1999)
enter the cell (Brul and Coote 1999). Reducing diffusional showed that MIC of lactic acid for B. cereus was 3.48 mg/
entry of the acid into the cell is probably a key mechanism mL, while MIC for Es. coli was 3.72 mg/mL. Some factors,
of yeasts’ resistance (Piper et al. 2001). Phenotypically such as bacterial strain and its sensitivity, inoculum volume,
acquired resistance to acid preservatives is well documented incubation time and temperature, affect the MIC and MBC
for yeasts, and this results from the enhanced ability of cell values (Zaika 2002; Raybaudi-Massila et al. 2009). More-
to catalyze energy-dependent extrusion acids (Warth 1985). over, the method used to assess the antimicrobial activity
Piper et al. (1998) showed evidence of an efflux system that could be related to the variations of the experimental results
removes accumulated anions from inside the cell. (Jiang 2011).
In comparison with other tested yeasts, Sac. cerevisiae was
the least sensitive to lactic acid. Strong inhibitory effects
Lactic Acid MIC, MBC and MFC
against Sac. cerevisiae require relatively high lactic acid con-
The results of lactic acid MICs, MBCs (for bacteria) and centration (Thomas et al. 2002; Abbott et al. 2008), which is
MICs, MFCs (for yeasts) against the tested microorganisms in agreement with our results. The inhibitory effect of lactic
are given in Table 3. Lactic acid MIC for bacteria ranged and acetic acid on the growth of Sac. cerevisiae was investi-

Journal of Food Processing and Preservation •• (2015) ••–•• © 2015 Wiley Periodicals, Inc. 5
ANTIMICROBIAL ACTIVITY OF LACTIC ACID S. STANOJEVIĆ-NIKOLIĆ ET AL.

gated by Narendranath et al. (2001). The results of this

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