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Article

The Influence of Environmental Conditions on the Antagonistic Activity of Lactic Acid Bacteria Isolated from Fermented Meat Products

by
Anna Łepecka
1,2,*,
Piotr Szymański
1,
Sylwia Rutkowska
2,
Kinga Iwanowska
2 and
Danuta Kołożyn-Krajewska
2
1
Department of Meat and Fat Technology, Prof. Waclaw Dabrowski Institute of Agriculture and Food Biotechnology—State Research Institute, Rakowiecka 36 St., 02-532 Warsaw, Poland
2
Department of Food Gastronomy and Food Hygiene, Institute of Human Nutrition Sciences, Warsaw University of Life Sciences—SGGW, Nowoursynowska 159c St., 02-776 Warsaw, Poland
*
Author to whom correspondence should be addressed.
Foods 2021, 10(10), 2267; https://doi.org/10.3390/foods10102267
Submission received: 18 August 2021 / Revised: 17 September 2021 / Accepted: 22 September 2021 / Published: 25 September 2021
(This article belongs to the Special Issue Application of Lactic Acid Bacteria in Food Preservation and Fermentation)
Browse Figures
Versions Notes

Abstract

:
The aim of this study was to determine the impact of environmental conditions on the antimicrobial properties of 21 lactic acid bacteria strains in the selected indicator bacteria. To assess the antimicrobial activity of the whole bacteria culture (WBC), the agar well diffusion method was used. The interference of LAB strains with the growth of the selected indicator bacteria was evaluated by incubating co-cultures in the food matrix. Based on the conducted research, it was found that environmental conditions have a significant impact on the antimicrobial activity of lactic acid bacteria strains. The highest antimicrobial activity was recorded under optimal conditions for the development of LAB, the incubation time being different depending on the indicator strain used. The tested LAB strains were characterized by a high ability to inhibit indicator strains, especially in the food matrix. These results led us to further characterize and purify the antimicrobial compound produced by lactic acid bacteria taking into account changing environmental conditions.

Graphical Abstract

1. Introduction

Lactic acid bacteria (LAB, LABs) exhibit antimicrobial properties against Gram(+) and Gram(-) bacteria. There are several mechanisms of antimicrobial action, and not all of them have been fully explained. These mechanisms can be competition for the adherence site, competition for nutrients, and/or production of non-specific antibacterial substances. LAB may cause the inactivation of other microorganisms, mainly through the production of lactic acid, acetic acid, propionic acid, diacetyl, acetaldehyde, H2O2, bacteriocins, exopolysaccharides, fatty acids, and amino acids metabolites [1,2]. Typically, bacteria produce more than one antimicrobial substance, which increases the spectrum of their activity [3]. LAB strains can function as microbial barriers against gastrointestinal pathogens through the competitive exclusion of pathogen binding, modulation of the host’s immune system, and production of inhibitory compounds [4]. In recent years there has been a search for new antimicrobial substances that are safe, natural and eco-friendly. LAB are very potential candidates because of their widely known positive healthy effects [5,6,7,8].
By securing favorable environmental conditions, LAB inhibit the development of pathogenic microflora. In order to understand the most favorable conditions for the production of antimicrobial substances, and to examine the impact of stress on their synthesis, a detailed analysis should be carried out, taking into account many factors. According to other studies [9,10], cell culture conditions deviating from the optimum increased the amount of produced antimicrobial compounds. Environmental stress can lead to the incorporation of defense mechanisms into LAB cells, and thereby intensify the production of protein substances which are antagonistic to other bacteria, in order to ensure the survival of cells under unfavorable conditions. Probably, a modification of environmental conditions may increase the effectiveness of the search for strains with strong antimicrobial properties. In fact, the antagonistic properties of individual microorganisms are difficult to assess. According to Mataragas et al. [11], the production of antimicrobial substances was strongly affected by the manipulation of temperature and pH. In order to effectively use antimicrobial compounds, it is necessary to optimize production conditions by controlling technological parameters and the environment. A detailed analysis of the biochemical and technological properties of these types of compounds can thus explain their low activity.
The aim of this study was to determine the impact of environmental conditions (incubation time, temperature, and pH) on the antimicrobial properties of LAB strains isolated from fermented meat products on the selected indicator bacteria and study the effects of LAB and pathogens against each other in experimental co-culture.

2. Materials and Methods

2.1. Strain Collection and Bacterial Growth Conditions

A total of 21 LAB strains were previously isolated from organic fermented meat products [12]. The list of LAB strains is presented below in Table 1.
The isolates were identified by the sequencing of 16S rRNA (Genbank accession were noted) and were tested for probiotic properties (survival studies, safety assessment, enzyme and antimicrobial activities). The strains come from a collection of the Department of Food Gastronomy and Food Hygiene WULS-SGGW. The foodborne commensal strains and pathogens were selected as indicator strains based on previous studies [14,15,16,17]. Escherichia coli ATCC 10536, Salmonella enteritidis ATCC 13076, Listeria monocytogenes ATCC 19111, Staphylococcus aureus ATCC 25923 and Bacillus cereus ATCC 14579 were used as indicators and were received from American Type Culture Collection (ATCC).
LAB strains were stored in MRS broth (de Man, Rogosa and Sharpe broth, LabM Ltd., Bury, UK) and the indicator strains were stored in nutrient broth (Nutrient broth No. 2, LabM Ltd., Bury, UK) at −80 °C with 20% glycerol (w/v, Merck KGaA, Darmstadt, Germany). LAB isolates and indicator bacteria were cultivated in MRS and nutrient broth, respectively, at 37 °C for 24 h under anaerobic conditions before each experiment. All experiments were performed in triplicate.

2.2. Antagonistic Activity

To assess the antimicrobial activity of whole bacteria culture (WBC), the agar well diffusion method was used according to Soleimani et al. and Ołdak et al. [15,18]. A 0.5 McFarland standard suspension of LAB strain in MRS broth with a turbidity of 1.5 × 108 CFU mL−1 was used to standardize the inoculum. Sterile Petri dishes with Mueller-Hinton agar I (MHA, LabM Ltd., Bury, UK) were inoculated with 200 µL of a 24 h culture of the indicator bacteria in nutrient broth (concentration ≈ 106 CFU mL−1). Wells with a diameter of 5.5 mm were then cut out and filled with 50 µL of WBC. The plates were incubated at 37 °C for 24 h and the inhibition zone diameter (X) was determined as follows:
X = D − d
where X—inhibition zone diameter (mm); D—total growth inhibition zone diameter (mm); d—the well diameter (5.5 mm).

2.3. Study of the Effect of Incubation Time on Antagonistic Activity

The effect of incubation time on the microbiological activity of LAB strains was tested at four time intervals (24, 48, 72, and 96 h) at a temperature of 37 °C [10]. The cultures were performed as described above in Section 2.2.

2.4. Study of the Effect of Temperature on Antagonistic Activity

The effect of temperature on the microbiological activity of LAB strains was tested at three temperatures. The strains were incubated in MRS broth under modified temperatures: 20, 30 and 37 °C for 24 h and then used for testing [9]. In addition, an experiment was carried out using strains subjected to stress conditions. The strains were first incubated at 37 °C for 24 h and then placed in a water bath at 55 or 100 °C during 10 min [9]. These temperatures are often considered critical for microbial growth and were selected on the basis of the cited publication. The cultures were performed as described above in Section 2.2.

2.5. Study of the Effect of pH on Antagonistic Activity

Strains were incubated for 24 h at 37 °C in nutrient broth with a different pH of 4.0, 5.7, 6.4, and 8.0 [9]. To obtain a pH of 4.0 and 5.7, concentrated 1M HCl (Merck KGaA, Darmstadt, Germany) was gradually added to the MRS broth. The pH of 6.4 was obtained by preparing the MRS broth according to the instructions on the preparation label. The pH of 8.0 was obtained by pipetting 1M NaOH (Merck KGaA, Darmstadt, Germany). The pH changes were measured using the Elmetron CP-511 pH-meter (Elmetron Sp.j., Zabrze, Poland). Then, the modified MRS broths were sterilized for 15 min at 121 °C. LAB strains were incubated in modified MRS broths for 24 h at 37 °C and then used for testing. The cultures were performed as described above in Section 2.2.

2.6. Co-Culture Assay

The interference of LAB strains with the growth of the selected indicator bacteria was evaluated by incubating a co-culture of both strains and comparing the recovered cells with those obtained from pure cultures of both strains. Based on the results of statistical analysis, two strains were selected for co-culture studies. The UHT milk was selected as the food matrix because of the pH (6.6) similar to the pH of the MRS broth (6.4; culture broth for LAB). Three indicator strains were selected, whose growth was mostly inhibited by the tested LAB (selection based on statistical analysis). The temperature of 37 °C was the optimal condition for the growth of most of the foodborne pathogens. The same time intervals indicated in the previous section were used.
Co-culture tests were performed according to Ołdak et al. [15] with slight modifications. In total, 50 mL of UHT milk was inoculated with the LAB strain to induce fermentation. A 0.5 McFarland standard suspension of LAB strain in MRS broth with a turbidity of 1.5 × 108 CFU mL−1 was used to standardize the inoculum. The milk was incubated for 24 h at 37 °C. Then, after 24 h, 1 mL of indicator bacteria in nutrient broth (concentration 106 CFU mL−1) was added to the fermented milk and placed at a temperature of 37 °C (RS—research sample). The following test designations were used: RS-LAB—research sample, Lb. plantarum SCH4 or BAL7; RS-Lm—research sample, L. monocytogenes ATCC 19111; RS-Ec—research sample, E. coli ATCC 10536; RS-Sa—research sample, S. aureus ATCC 25923. The samples inoculated only with the indicator strains or only with the Lb. plantarum strains were also prepared as controls (CS—control sample). The following test designations were used: CS-LAB—control sample, Lb. plantarum SCH4 or BAL7; CS-Lm—control sample, L. monocytogenes ATCC 19111; CS-Ec—control sample, E. coli ATCC 10536; CS-Sa—control sample, S. aureus ATCC 25923. In the case of the incubation of the milk with the indicator bacteria, no fermentation of milk was carried out. Microbiological cultures were performed at time 0 and after 24, 48, 72, and 96 h of the incubation.
LAB were plated on MRS agar (de Man, Rogosa and Sharpe agar, LabM Ltd., Bury, UK), L. monocytogenes was plated on PALCAM agar (LabM Ltd., Bury, UK), and E. coli was plated on TBX agar (Tryptone Bile X-Glucuronide Agar, LabM Ltd., Bury, UK), S. aureus was plated on Baird-Parker agar with Egg Yolk Tellurite Supplement (LabM Ltd., Bury, UK). The plates with MRS agar, Baird-Parker agar, and PALCAM agar were incubated at 37 °C for 24–48 h and the plates with TBX agar were incubated at 44 °C for 24 h. After incubation, colonies on each solid medium were counted and the results obtained were transformed into a logarithmic scale. The number of LAB and the number of indicator bacteria were checked in parallel. The results were presented on linear charts.

2.7. Statistical Analysis

The test results were given after averaging the results from three replicates of the zones of the inhibition areas. The standard deviation was calculated. A one-way analysis of variance (ANOVA) and post-hoc analyses (Tukey’s HSD) were performed at the significance level of p < 0.05. The statistical results were analyzed in STATISTICA 13 (TIBCO Software Inc., Palo Alto, California, USA) program.

3. Results and Discussion

Antimicrobial activity may be affected by several factors, including the pH value of the environment, temperature, cell growth phase, co-culture incubation, culture medium composition, aerobic conditions, presence of surface tension reducing agents, and the addition of mineral salts [9,19,20,21].
Figure 1 presents the impact of incubation time on the antimicrobial activity of LAB strains. Based on the obtained results, it was found that the incubation time had a significant effect on the antimicrobial properties of the strains (p < 0.05), but the use of an indicator microorganism was also important. The highest inhibition of E. coli (p < 0.05) was observed after 72 h. In the case of S. enteritidis, significantly different growth inhibition was observed after 24 and 48 h. The greatest inhibition of L. monocytogenes growth was observed after 24 and 72 h (p < 0.05). With respect to S. aureus, a significant inhibition of bacterial growth was observed after 48 h, and inhibition of B. cereus after 24 h. Extending the incubation time to 96 h resulted in a significant decrease in antagonistic activity.
Bacteria are affected by the surrounding environment and they react dynamically to changes in external conditions. The number of cells in the culture medium determines the antagonistic activity of the lactic bacteria. The greatest synthesis of microbial agents occurs in the terminal phase of logarithmic growth and at the beginning of the stationary phase [22]. The logarithmic growth phase is characterized by the most intensive cell growth at a constant rate. The duration of this phase is determined by several factors, such as the amount of nutrients, the presence of toxic substances, cell metabolism products, pH value, temperature, oxygen content and type of culture. On the other hand, the stationary phase is characterized by a fixed number of cells, which is the result of the balance between the number of live and dead cells. Not always is the amount of biomass produced coupled with the production of microbial agents. Depending on the incubation temperature of the strain, the synthesis may occur earlier or later [10]. It has been reported that stress responses vary depending on the logarithmic growth phase of LAB, i.e., cells in the stationary phase develop more general resistance to various types of stresses compared to cells in the exponential growth phase [23,24]. The effect of different cultivation conditions, in particular incubation time, carbon, nitrogen and mineral sources, on EPS (Exopolysaccharides) production often varies from strain to strain [25].
Figure 2 shows the effect of temperature on the antimicrobial activity of LAB strains. This research showed that the statistically significant inhibition areas of the indicator bacteria growth were shown at 37 °C (p < 0.05). At 20 and 30 °C temperatures, a slight inhibition of the indicator bacteria growth was observed. Interestingly, LAB strains subjected to stress conditions (55 and 100 °C) still showed quite strong antimicrobial activity. The strongest antimicrobial properties at 55 and 100 °C were found against B. cereus ATCC 14579 and E. coli ATCC 10536, respectively. The lowest, not statistically different, antagonistic properties were found at 20 °C and subjected to short-term exposure at 100 °C. The question, therefore, arises whether dead cell metabolites also have antagonistic properties [26,27,28]. Understanding the response to adverse temperature conditions is a task that requires the consideration of many factors that could modulate bacterial behavior. Therefore, studies about the effects of different incubation temperatures on cell survival should be extended to studies on adaptability to moderate stresses, which may increase resistance to more adverse conditions, and molecular studies of changes in the profile of produced proteins are needed. High temperatures may induce the production of various types of proteins, such as heat shock proteins (HSPs) [29].
The growth temperature for mesophilic LAB is 15–45 °C [30]. A temperature of 37 °C is optimal for the development of most human pathogenic bacteria. The optimal temperature for LAB development does not always coincide with the temperature where the synthesis of microbial agents is the most intensive. Firstly, lower temperatures favor lower LAB biomass production, which may favor the production of non-specific metabolites [31]. On the other hand, at a higher temperature, more energy can be retained in the cell, which in turn facilitates the synthesis of antimicrobial agents [19]. High temperatures can also have a beneficial effect on metabolite production. Bacteria exposed to stress increase the production of protective factors to preserve vitality and metabolic functions, thereby determining the production of microbial agents. In many cases, bacteriocins are the most intensively produced by LAB at the optimum temperature for their growth [22,27,28,29,30,31,32,33,34,35,36]. However, this study also presents cases when the environmental temperature exceeds the optimum triggered the defense mechanisms of cells of other LAB species, which was associated with increased production of compounds with antagonistic properties.
The impact of pH on the antimicrobial activity of LAB strains is shown in Figure 3. The tested strains have the strongest antimicrobial properties at pH of 6.4 against all of the tested microorganisms with the strongest antimicrobial activity against L. monocytogenes ATCC 19111 (p < 0.05). A change in the pH of the environment below 6.4 resulted in a decrease or total disappearance of the antimicrobial properties of strains. Lowering the pH to 5.7 value significantly reduced the created zones of inhibition. On the other hand, a further decrease in the pH of the environment to 4.0 and an increase outside the optimum to 8.0 resulted in a decrease in antagonist activity at the same level. A decrease in the antimicrobial activity of CFS of Lb. casei, Lb. acidophilus, and Lb. salivarius strains at a pH above 7 was observed by Moradi et al. [37]. A high percentage of residual antimicrobial activity against L. monocytogenes was observed at a pH of 4.0 and 5.0.
LAB strains have optimum pH environmental conditions in the range of 5.4–6.4. However, it has been found that the isolates of this species have survived even at a pH of 2.8 or 10.0 [38]. Thanks to their ability to regulate intracellular pH, they can survive in a relatively low pH environment (low concentration of hydrogen ions damages the natural mechanism of pH gradient). As the enzymatic reactions are controlled by pH value, the decrease of pH may have positive effects on the synthesis of antimicrobial compounds, such as bacteriocins, through increasing the pools of essential metabolites, including ATP. However, enzymatic reactions are regulated by pH and temperature, which in consequence can lead to their inhibition and accordingly to the inhibition of the synthesis of antimicrobial compounds [38].
Several studies [39,40,41,42] have been carried out on the application of the pure culture of LAB to control the growth of foodborne microorganisms. According to the statistical analysis, Lb. plantarum SCH4 and BAL7, L. monocytogenes ATCC 19111, E. coli ATCC 10536, and S. aureus ATCC 25923 were selected for a further study. The relationship between strains and environmental conditions are presented in Figure 4a–f.
Co-culture is a system containing two distinct types of cells and allowing the growth of both types of microorganisms [43]. Tabasco et al. [44] reported that antimicrobial compound synthesis and enhanced antimicrobial properties may occur, provided that bacteria are used in co-culture with starter microorganisms (e.g., starter yogurt culture Str. thermophilus and Lb. delbrueckii subsp. bulgaricus), or in co-culture with other Gram-positive and Gram-negative bacteria strains or the presence of the supernatant from their culture. The competition can have a positive impact on the production of protein substances.
Figure 4a–c shows the effect of Lb. plantarum SCH4 on the viability of indicator strains. As a result of this study, a statistically significant inhibitory effect on L. monocytogenes ATCC 19111, E. coli ATCC 10536, and S. aureus ATCC 25923 was observed after 72 h of incubation. Compared to control sample (CS-Lm), a decrease in the number of L. monocytogenes ATCC 19111 was observed after 72 h (1 log CFU mL−1) and after 96 h (5 log CFU mL−1) of incubation. In the case of E. coli ATCC 10536 a non-statistically significant increase in the number of indicator strains was observed after 24 h and a decrease in number after 48 h. After 72 h, a statistically significant (p < 0.05) decrease in the number of E. coli ATCC 10536 was noted (about 2 log CFU mL−1) compared to the initial number (about 6 log CFU mL−1). After 96 h, the number of E. coli ATCC 10536 cells decreased to 3.00 log CFU mL−1. In control sample (CS-Ec), a significant increase was observed (8 log CFU mL−1). The number of S. aureus ATCC 25923 significantly (p < 0.05) decreased after 72 h (3.00 log CFU mL−1), and after 96 h (1.00 log CFU mL−1). The number of bacteria in control sample (CS-Sa) increased steadily and after 96 h was almost 7 log CFU mL−1. The number of viable cells of Lb. plantarum SCH4 in control sample (CS-LAB) and research sample (RS-LAB) throughout the incubation was 8–9 log CFU mL−1. In summary, the significant effect of Lb. plantarum SCH4 on inhibiting the indicator strain growth in the food matrix was demonstrated.
The Lb. plantarum BAL7 strain showed even more strong antimicrobial activity (Figure 4d–f). A statistically significant (p < 0.05) decrease in the number of L. monocytogenes ATCC 19111 was observed after 48 and 72 h (to 4.00 log CFU mL−1), and the complete inhibition of growth was observed after 96 h. The number of viable cells of L. monocytogenes ATCC 19111 in the control sample (CS-Lm) after 48, 72, and 96 h was about 9 log CFU mL−1. In the case of E. coli ATCC 10536, a significant decrease in bacterial numbers was observed after 72 and 96 h (≈5 and 4 log CFU mL−1, respectively). The number of E. coli ATCC 10536 in the control sample (CS-Ec) also decreased, but to a lesser extent than in the test sample (RS-Ec). In contrast, the largest inhibitory effect of Lb. plantarum BAL7 strain was in relation to S. aureus ATCC 25923. A statistically significant (p < 0.05) decrease in the number of bacterial cells was observed after 48 h. After 72 h, the growth of S. aureus ATCC 25923 cells was completely inhibited. In the control (CS-Sa), a reduction in the number of S. aureus ATCC 25923 was observed after 24 h, followed by a bacterial cell count of ≈4 log CFU mL−1 for the remaining incubation time. The Lb. plantarum BAL7 strain was characterized by the high ability to inhibit the indicator bacteria growth in the food matrix.
The capability of the Lactobacillus isolates to inhibit the growth of S. typhi was evaluated in a co-culture experiment by Abdel-Daim et al. [4]. The results showed that nine Lactobacillus strains inhibited the growth of Salmonella after 24 h of incubation. However, the growth of tested Lactobacillus isolates was not affected by the simultaneous presence of Salmonella typhi. Similar test results were obtained by Ołdak et al. [15], where Lb. plantarum strains isolated from regional cheeses, showed the greatest antimicrobial activity after 24 and 64 h of incubation in co-cultures. Based on our co-culture results, a statistically significant inhibition of indicator strain growth was achieved after 48 or 72 h, depending on the type of the strain tested. However, it should be noted that the high-dose inoculum of indicator strains was used (~106 CFU mL−1). In the study of Shah et al. [43] it was observed that in the co-culture, LAB strains showed a significant reduction of growth of E. coli after 24 h of incubation. Similar to our findings, Kang et al. [45] had also observed that Lb. salivarius and Lb. fermentum strains led to a significant reduction in S. aureus, but after 24 h. Koohestani et al. [46] showed that the application of CFS of Lactobacillus strains as antibacterial and biofilm removal compounds could be very suitable to control the growth of foodborne pathogens. Antimicrobial properties, production of bacteriocins or other antimicrobial substances can have potential preserving natural food. In order to increase the productivity of these substances, it is necessary to know the optimal conditions for their production. The production of antimicrobial substances, including bacteriocins, is strictly dependent on conditions such as environmental pH, temperature and incubation time. These conditions may differ from precisely established ones and be optimal for a given species of bacteria [47]. These factors could affect the viability and functional properties of microorganisms. Defense mechanisms against stress conditions were developed by LAB in order to survive when exposed to sudden environmental changes [48].

4. Conclusions

According to Parlindungan et al. [49] due to the growing interest in probiotics, it becomes crucial to characterize new probiotic strains with unique probiotic properties. LAB strains isolated from fermented meat products showed a strong activity towards indicator bacteria. The experiments in this study were conducted both on the microbiological medium and the food matrix. Based on our results, it was found that the largest inhibitory effect of Lb. plantarum strains on indicator bacteria were observed after 24 and 72 h of the incubation, at 37 °C and with a pH of 6.4. The greatest synthesis of microbial agents occurs in the terminal phase of logarithmic growth and at the beginning of the stationary phase. The temperature of 37 °C and pH of 6.4 are considered optimal for the growth of LAB and it was under these conditions that the greatest antimicrobial properties were found. Antimicrobial properties have also been observed in suboptimal conditions, indicating the potential for the production of antimicrobial substances under stress conditions.
This study will contribute to a better understanding of the influence of environmental factors on the behavior of the LAB strains. The tested isolates with potential probiotic properties showed promising antagonistic activity against some indicator strains. As antimicrobial activity is strain-dependent, more thorough research into compounds with antimicrobial properties produced by the studied strains is needed.
The results of this study indicate that LAB are capable of exhibiting antimicrobial activity under various environmental conditions, both optimal and non-optimal, which indicates the potential for the production of some antimicrobial substances. A modification of environmental conditions may increase the effectiveness of the search for strains with strong antimicrobial properties.

Author Contributions

Conceptualization, A.Ł.; methodology, A.Ł.; formal analysis, A.Ł., D.K.-K.; investigation, A.Ł., S.R., K.I.; writing—original draft preparation, A.Ł, P.S.; writing—review and editing, P.S., D.K.-K.; visualization, A.Ł.; supervision, D.K.-K. All authors have read and agreed to the published version of the manuscript.

Funding

Research partially financed by the Polish Ministry of Science and Higher Education within funds of the Institute of Human Nutrition Sciences, Warsaw University of Life Sciences (WULS), for scientific research.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

Not applicable.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Bajpai, V.K.; Han, J.H.; Rather, I.A.; Park, C.; Lim, J.; Paek, W.K.; Lee, J.S.; Park, Y.H. Characterization and antibacterial potential of lactic acid bacterium Pediococcus pentosaceus 4I1 isolated from freshwater fish Zacco koreanus. Front. Microbiol. 2016, 7, 2037. [Google Scholar] [CrossRef] [Green Version]
  2. Ratajczak, K.; Piotrowska-Cyplik, A. Metabolites of lactic acid bacteria—Overview and industrial applications. Adv. Microbiol. 2017, 56, 416–421. [Google Scholar] [CrossRef]
  3. Papadimitriou, K.; Zoumpopoulou, G.; Foligné, B.; Alexandraki, V.; Kazou, M.; Pot, B.; Tsakalidou, E. Discovering probiotic microorganisms: In vitro, in vivo, genetic and omics approaches. Front. Microbiol. 2015, 6, 58. [Google Scholar] [CrossRef] [Green Version]
  4. Abdel-Daim, A.; Hassouna, N.; Hafez, M.; Ashor, M.S.A.; Aboulwafa, M.M. Antagonistic activity of Lactobacillus isolates against Salmonella typhi in vitro. BioMed Res. Int. 2013, 2013, 680605. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  5. Fontana, C.; Cocconcelli, P.S.; Vignolo, G.; Saavedra, L. Occurrence of antilisterial structural bacteriocins genes in meat borne lactic acid bacteria. Food Control 2015, 47, 53–59. [Google Scholar] [CrossRef]
  6. Jabbari, V.; Khiabani, M.S.; Mokarram, R.R.; Hassanzadeh, A.M.; Ahmadi, E.; Gharenaghadeh, S.; Karimi, N.; Kafil, H.S. Lactobacillus plantarum as a probiotic potential from kouzeh cheese (traditional Iranian cheese) and its antimicrobial activity. Probiotics Antimicrob. Proteins 2017, 9, 189–193. [Google Scholar] [CrossRef] [PubMed]
  7. Shi, Y.; Cui, X.; Gu, S.; Yan, X.; Li, R.; Xia, S.; Chen, H.; Ge, J. Antioxidative and probiotic activities of lactic acid bacteria isolated from traditional artisanal milk cheese from Northeast China. Probiotics Antimicrob. Proteins 2019, 11, 1086–1099. [Google Scholar] [CrossRef] [PubMed]
  8. Trush, E.A.; Poluektova, E.A.; Beniashvilli, A.G.; Shifrin, O.S.; Poluektov, Y.M.; Ivashkin, V.T. The Evolution of Human Probiotics: Challenges and Prospects. Probiotics Antimicrob. Proteins 2020, 12, 1291–1299. [Google Scholar] [CrossRef]
  9. Ogunbanwo, S.T.; Sanni, A.I.; Onilude, A.A. Influence of cultural conditions on the production of bacteriocin by Lactobacillus brevis OG1. Afr. J. Biotechnol. 2003, 2, 179–184. [Google Scholar] [CrossRef] [Green Version]
  10. Barbosa, M.S.; Todorov, S.D.; Ivanova, I.V.; Belguesmia, Y.; Choiset, Y.; Rabesona, H.; Chobert, J.M.; Haertle, T.; Franco, B.D.G.M. Characterization of a two-peptide plantaricin produced by Lactobacillus plantarum MBSa4 isolated from Brazilian salami. Food Control 2016, 60, 103–112. [Google Scholar] [CrossRef] [Green Version]
  11. Mataragas, M.; Metaxopoulos, J.; Galiotou, M.; Drosinos, E.H. Influence of pH and temperature on growth and bacteriocin production by Leuconostoc mesenteroides L124 and Lactobacillus curvatus L442. Meat Sci. 2003, 64, 265–271. [Google Scholar] [CrossRef]
  12. Rzepkowska, A.; Zielińska, D.; Ołdak, A.; Kołożyn-Krajewska, D. Safety assessment and antimicrobial properties of the lactic acid bacteria strains isolated from Polish raw fermented meat products. Int. J. Food Prop. 2017, 20, 2736–2747. [Google Scholar] [CrossRef] [Green Version]
  13. Zheng, J.; Wittouck, S.; Salvetti, E.; Franz, C.M.; Harris, H.; Mattarelli, P.; O’Toole, P.W.; Pot, B.; Vandamme, P.; Walter, J.; et al. A taxonomic note on the genus Lactobacillus: Description of 23 novel genera, emended description of the genus Lactobacillus Beijerinck 1901, and union of Lactobacillaceae and Leuconostocaceae. Int. J. Syst. Evol. Microbiol. 2020, 70, 2782–2858. [Google Scholar] [CrossRef] [PubMed]
  14. Lin, C.M.; Chu, Y.C.; Hsiao, C.P.; Wu, J.S.; Hsieh, C.W.; Hou, C.Y. The Optimization of Plasma-Activated Water Treatments to Inactivate Salmonella Enteritidis (ATCC 13076) on Shell Eggs. Foods 2019, 8, 520. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  15. Ołdak, A.; Zielińska, D.; Rzepkowska, A.; Kołożyn-Krajewska, D. Comparison of antibacterial activity of Lactobacillus plantarum strains isolated from two different kinds of regional cheeses from Poland: Oscypek and Korycinski Cheese. BioMed Res. Int. 2017, 2017, 6820369. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  16. Lindbäck, T.; Granum, P.E.  Bacillus cereus. In Food Microbiology: Fundamentals and Frontiers, 1st ed.; Doyle, M.P., Diez-Gonzalez, F., Hill, C., Eds.; ASM Press: Washington, DC, USA, 2019; pp. 541–554. [Google Scholar] [CrossRef]
  17. Wan, M.L.Y.; Forsythe, S.J.; El-Nezami, H. Probiotics interaction with foodborne pathogens: A potential alternative to antibiotics and future challenges. Crit. Rev. Food Sci. Nutr. 2019, 59, 3320–3333. [Google Scholar] [CrossRef]
  18. Soleimani, N.A.; Kermanshahi, R.K.; Yakhchali, B.; Sattari, T.N. Antagonistic activity of probiotic lactobacilli against Staphylococcus aureus isolated from bovine mastitis. Afr. J. Microbiol. Res. 2010, 4, 2169–2173. [Google Scholar] [CrossRef]
  19. Settanni, L.; Valmorri, S.; Suzzi, G.; Corsetti, A. The role of environmental factors and medium composition on bacteriocin-like inhibitory substances (BLIS) production by Enterococcus mundtii strains. Food Microbiol. 2008, 25, 722–728. [Google Scholar] [CrossRef]
  20. Smetanková, J.; Hladíková, Z.; Valach, F.; Zimanová, M.; Kohajdová, Z.; Greif, G.; Greifová, M. Influence of aerobic and anaerobic conditions on the growth and metabolism of selected strains of Lactobacillus plantarum. Acta Chim. Slovaca 2012, 5, 204–210. [Google Scholar] [CrossRef] [Green Version]
  21. Yang, J.; Wang, J.; Yang, K.; Liu, M.; Qi, Y.; Zhang, T.; Fan, M.; Wei, X. Antibacterial activity of selenium-enriched lactic acid bacteria against common food-borne pathogens in vitro. J. Dairy Sci. 2018, 101, 1930–1942. [Google Scholar] [CrossRef] [Green Version]
  22. Gong, H.S.; Meng, X.C.; Wang, H. Plantaricin MG active against Gram-negative bacteria produced by Lactobacillus plantarum KLDS1. 0391 isolated from “Jiaoke”, a traditional fermented cream from China. Food Control 2010, 21, 89–96. [Google Scholar] [CrossRef]
  23. Saarela, M.; Rantala, M.; Hallamaa, K.; Nohynek, L.; Virkajärvi, I.; Mättö, J. Stationary-phase acid and heat treatments for improvement of the viability of probiotic lactobacilli and bifidobacteria. J. Appl. Microbiol. 2004, 96, 1205–1214. [Google Scholar] [CrossRef] [PubMed]
  24. Settachaimongkon, S.; van Valenberg, H.J.; Gazi, I.; Nout, M.R.; van Hooijdonk, T.C.; Zwietering, M.H.; Smid, E.J. Influence of Lactobacillus plantarum WCFS1 on post-acidification, metabolite formation and survival of starter bacteria in set-yoghurt. Food Microbiol. 2016, 59, 14–22. [Google Scholar] [CrossRef] [PubMed]
  25. Mıdık, F.; Tokatlı, M.; Elmacı, S.B.; Özçelik, F. Influence of different culture conditions on exopolysaccharide production by indigenous lactic acid bacteria isolated from pickles. Arch. Microbiol. 2020, 202, 875–885. [Google Scholar] [CrossRef] [PubMed]
  26. Chen, Y.T.; Hsieh, P.S.; Ho, H.H.; Hsieh, S.H.; Kuo, Y.W.; Yang, S.F.; Lin, C.W. Antibacterial activity of viable and heat-killed probiotic strains against oral pathogens. Lett. Appl. Microbiol. 2020, 70, 310–317. [Google Scholar] [CrossRef]
  27. Tareb, R.; Bernardeau, M.; Gueguen, M.; Vernoux, J.P. In vitro characterization of aggregation and adhesion properties of viable and heat-killed forms of two probiotic Lactobacillus strains and interaction with foodborne zoonotic bacteria, especially Campylobacter jejuni. J. Med. Microbiol. 2013, 62, 637–649. [Google Scholar] [CrossRef]
  28. Zielińska, D.; Łepecka, A.; Ołdak, A.; Długosz, E.; Kołożyn-Krajewska, D. Growth and adhesion inhibition of pathogenic bacteria by live and heat-killed food-origin Lactobacillus strains or their supernatants. FEMS Microbiol. Lett. 2021, 368, fnab024. [Google Scholar] [CrossRef]
  29. Arena, M.P.; Capozzi, V.; Longo, A.; Russo, P.; Rieu, A.; Guzzo, J.; Fiocco, D. The phenotypic analysis of Lactobacillus plantarum shsp mutants reveals a potential role for hsp1 in cryotolerance. Front. Microbiol. 2019, 10, 838. [Google Scholar] [CrossRef]
  30. Todorov, S.D.; Franco, B.D.G.D.M. Lactobacillus plantarum: Characterization of the species and application in food production. Food Rev. Int. 2010, 26, 205–229. [Google Scholar] [CrossRef]
  31. Liu, S.; Ma, Y.; Zheng, Y.; Zhao, W.; Zhao, X.; Luo, T.; Zhang, J.; Yang, Z. Cold-stress response of probiotic Lactobacillus plantarum K25 by iTRAQ proteomic analysis. J. Microbiol. Biotechnol. 2020, 30, 187–195. [Google Scholar] [CrossRef] [PubMed]
  32. Aoudia, N.; Rieu, A.; Briandet, R.; Deschamps, J.; Chluba, J.; Jego, G.; Garrido, C.; Guzzo, J. Biofilms of Lactobacillus plantarum and Lactobacillus fermentum: Effect on stress responses, antagonistic effects on pathogen growth and immunomodulatory properties. Food Microbiol. 2016, 53, 51–59. [Google Scholar] [CrossRef]
  33. Duar, R.M.; Lin, X.B.; Zheng, J.; Martino, M.E.; Grenier, T.; Pérez-Muñoz, M.E.; Leulier, F.; Gänzle, M.; Walter, J. Lifestyles in transition: Evolution and natural history of the genus Lactobacillus. FEMS Microbiol. Rev. 2017, 41, S27–S48. [Google Scholar] [CrossRef] [Green Version]
  34. Hosseini Nezhad, M.; Hussain, M.A.; Britz, M.L. Stress responses in probiotic Lactobacillus casei. Crit. Rev. Food Sci. 2015, 55, 740–749. [Google Scholar] [CrossRef] [PubMed]
  35. Grosu-Tudor, S.S.; Brown, L.; Hebert, E.M.; Brezeanu, A.; Brinzan, A.; Fadda, S.; Mozzi, F.; Zamfir, M. S-layer production by Lactobacillus acidophilus IBB 801 under environmental stress conditions. Appl. Microbiol. Biotechnol. 2016, 100, 4573–4583. [Google Scholar] [CrossRef] [PubMed]
  36. Zotta, T.; Parente, E.; Ricciardi, A. Aerobic metabolism in the genus Lactobacillus: Impact on stress response and potential applications in the food industry. J. Appl. Microbiol. 2017, 122, 857–869. [Google Scholar] [CrossRef] [Green Version]
  37. Moradi, M.; Mardani, K.; Tajik, H. Characterization and application of postbiotics of Lactobacillus spp. on Listeria monocytogenes in vitro and in food models. LWT—Food Sci. Technol. 2019, 111, 457–464. [Google Scholar] [CrossRef]
  38. Hammes, W.P.; Hertel, C.; Genus, I.  Lactobacillus. In Bergey’s Manual of Systematic Bacteriology, 2nd ed.; Vos, P., Garrity, G., Jones, D., Krieg, N.R., Ludwig, W., Rainey, F.A., Schleifer, K.-H., Whitman, W., Eds.; Springer: New York, NY, USA, 2009; Volume 3, pp. 465–510. [Google Scholar] [CrossRef]
  39. Sharma, C.; Singh, B.P.; Thakur, N.; Gulati, S.; Gupta, S.; Mishra, S.K.; Panwar, H. Antibacterial effects of Lactobacillus isolates of curd and human milk origin against food-borne and human pathogens. 3 Biotech. 2017, 7, 31. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  40. Hu, C.H.; Ren, L.Q.; Zhou, Y.; Ye, B.C. Characterization of antimicrobial activity of three Lactobacillus plantarum strains isolated from Chinese traditional dairy food. Food Sci. Nutr. 2019, 7, 1997–2005. [Google Scholar] [CrossRef] [Green Version]
  41. Ołdak, A.; Zielińska, D.; Łepecka, A.; Długosz, E.; Kołożyn-Krajewska, D. Lactobacillus plantarum Strains Isolated from Polish Regional Cheeses Exhibit Anti-Staphylococcal Activity and Selected Probiotic Properties. Probiotics Antimicrob. Proteins 2019, 12, 1025–1038. [Google Scholar] [CrossRef] [Green Version]
  42. Eddine, S.D.; Yasmine, S.; Fatima, G.; Amina, Z.; Battache, G.; Mebrouk, K. Antifungal and antibacterial activity of some lactobacilli isolated from camel’s milk biotope in the south of Algeria. J. Microbiol. Biotechnol. Food Sci. 2020, 9, 871–877. [Google Scholar] [CrossRef]
  43. Shah, N.; Patel, A.; Ambalam, P.; Holst, O.; Ljungh, A.; Prajapati, J. Determination of an antimicrobial activity of Weissella confusa, Lactobacillus fermentum, and Lactobacillus plantarum against clinical pathogenic strains of Escherichia coli and Staphylococcus aureus in co-culture. Ann. Microbiol. 2016, 66, 1137–1143. [Google Scholar] [CrossRef]
  44. Tabasco, R.; García-Cayuela, T.; Peláez, C.; Requena, T. Lactobacillus acidophilus La-5 increases lactacin B production when it senses live target bacteria. Int. J. Food Microbiol. 2009, 132, 109–116. [Google Scholar] [CrossRef]
  45. Kang, M.S.; Lim, H.S.; Oh, J.S.; Lim, Y.J.; Wuertz-Kozak, K.; Harro, J.M.; Shirtliff, M.E.; Achermann, Y. Antimicrobial activity of Lactobacillus salivarius and Lactobacillus fermentum against Staphylococcus aureus. Path. Dis. 2017, 75, ftx009. [Google Scholar] [CrossRef]
  46. Koohestani, M.; Moradi, M.; Tajik, H.; Badali, A. Effects of cell-free supernatant of Lactobacillus acidophilus LA5 and Lactobacillus casei 431 against planktonic form and biofilm of Staphylococcus aureus. Vet. Res. Forum 2018, 9, 301–306. [Google Scholar] [CrossRef]
  47. Aasen, I.M.; Møretrø, T.; Katla, T.; Axelsson, L.; Storrø, I. Influence of complex nutrients, temperature and pH on bacteriocin production by Lactobacillus sakei CCUG 42687. Appl. Microbiol. Biotechnol. 2000, 53, 159–166. [Google Scholar] [CrossRef]
  48. Ferrando, V.; Quiberoni, A.; Reinheimer, J.; Suárez, V. Functional properties of Lactobacillus plantarum strains: A study in vitro of heat stress influence. Food Microbiol. 2016, 54, 154–161. [Google Scholar] [CrossRef]
  49. Parlindungan, E.; Lugli, G.A.; Ventura, M.; van Sinderen, D.; Mahony, J. Lactic Acid Bacteria Diversity and Characterization of Probiotic Candidates in Fermented Meats. Foods 2021, 10, 1519. [Google Scholar] [CrossRef] [PubMed]
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Figure 1. Heatmap about the influence of incubation time on the antimicrobial activity of LAB strains and against indicator bacteria (n = 3). Means in the same column followed by different letters (A–D) represent significant differences (p < 0.05) for each incubation time and indicator bacteria.
Figure 1. Heatmap about the influence of incubation time on the antimicrobial activity of LAB strains and against indicator bacteria (n = 3). Means in the same column followed by different letters (A–D) represent significant differences (p < 0.05) for each incubation time and indicator bacteria.
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Foods 10 02267 g002 550
Figure 2. Heatmap about the influence of temperature on the antimicrobial activity of LAB strains and against indicator bacteria (n = 3). Means in the same column followed by different letters (A–D) represent significant differences (p < 0.05) for each temperature and indicator bacteria.
Figure 2. Heatmap about the influence of temperature on the antimicrobial activity of LAB strains and against indicator bacteria (n = 3). Means in the same column followed by different letters (A–D) represent significant differences (p < 0.05) for each temperature and indicator bacteria.
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Foods 10 02267 g003 550
Figure 3. Heatmap about the influence of pH on the antimicrobial activity of LAB strains and against indicator bacteria (n = 3). Means in the same column followed by different letters (A–D) represent significant differences (p < 0.05) for each temperature and indicator bacteria.
Figure 3. Heatmap about the influence of pH on the antimicrobial activity of LAB strains and against indicator bacteria (n = 3). Means in the same column followed by different letters (A–D) represent significant differences (p < 0.05) for each temperature and indicator bacteria.
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Foods 10 02267 g004 550
Figure 4. (a) Impact of Lb. plantarum SCH4 on the viability of L. monocytogenes ATCC 19111 in milk; (b) impact of Lb. plantarum SCH4 on the viability E. coli ATCC 10536 in milk; (c) impact of Lb. plantarum SCH4 on the viability of S. aureus ATCC 25923 in milk; (d) impact of Lb. plantarum BAL7 on the viability of L. monocytogenes ATCC 19111 in milk; (e) impact of Lb. plantarum BAL7 on the viability of E. coli ATCC 10536 in milk; (f) impact of Lb. plantarum BAL7 on the viability of S. aureus ATCC 25923 in milk; explanatory: CS-LAB—control sample, Lb. plantarum SCH4 or BAL7; RS-LAB—research sample, Lb. plantarum SCH4 or BAL7; CS-Lm—control sample, L. monocytogenes ATCC 19111; RS-Lm—research sample, L. monocytogenes ATCC 19111; CS-Ec—control sample, E. coli ATCC 10536; RS-Ec—research sample, E. coli ATCC 10536; CS-Sa—control sample, S. aureus ATCC 25923; RS-Sa—research sample, S. aureus ATCC 25923; the graph shows the log bacterial cell count (CFU mL−1) with standard deviation. Means in the same row followed by different uppercase letters within the same sample in different times are significantly different (p < 0.05); means in the same column followed by different lowercase letters within samples in the same time are significantly different (p < 0.05).
Figure 4. (a) Impact of Lb. plantarum SCH4 on the viability of L. monocytogenes ATCC 19111 in milk; (b) impact of Lb. plantarum SCH4 on the viability E. coli ATCC 10536 in milk; (c) impact of Lb. plantarum SCH4 on the viability of S. aureus ATCC 25923 in milk; (d) impact of Lb. plantarum BAL7 on the viability of L. monocytogenes ATCC 19111 in milk; (e) impact of Lb. plantarum BAL7 on the viability of E. coli ATCC 10536 in milk; (f) impact of Lb. plantarum BAL7 on the viability of S. aureus ATCC 25923 in milk; explanatory: CS-LAB—control sample, Lb. plantarum SCH4 or BAL7; RS-LAB—research sample, Lb. plantarum SCH4 or BAL7; CS-Lm—control sample, L. monocytogenes ATCC 19111; RS-Lm—research sample, L. monocytogenes ATCC 19111; CS-Ec—control sample, E. coli ATCC 10536; RS-Ec—research sample, E. coli ATCC 10536; CS-Sa—control sample, S. aureus ATCC 25923; RS-Sa—research sample, S. aureus ATCC 25923; the graph shows the log bacterial cell count (CFU mL−1) with standard deviation. Means in the same row followed by different uppercase letters within the same sample in different times are significantly different (p < 0.05); means in the same column followed by different lowercase letters within samples in the same time are significantly different (p < 0.05).
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Table
Table 1. The tested LAB strains and new nomenclature of LAB.
Table 1. The tested LAB strains and new nomenclature of LAB.
Tested LAB Strains [12]GenBank Accession [12]New Nomenclature of LAB [13]
Lactobacillus plantarum SCH1KX 014848 Lactiplantibacillus plantarum SCH1
Lactobacillus plantarum SCH2KX 014847Lactiplantibacillus plantarum SCH2
Lactobacillus plantarum SCH3KX 021365Lactiplantibacillus plantarum SCH3
Lactobacillus plantarum SCH4KX 021366 Lactiplantibacillus plantarum SCH4
Lactobacillus plantarum BAL7KX 021348Lactiplantibacillus plantarum BAL7
Lactobacillus brevis SCH5KX 021367Levilactobacillus brevis SCH5
Lactobacillus brevis SCH6KX 021368 Levilactobacillus brevis SCH6
Lactobacillus brevis BAL1KX 021369 Levilactobacillus brevis BAL1
Lactobacillus brevis BAL10KX 021350 Levilactobacillus brevis BAL10
Lactobacillus brevis KL3KX 021351 Levilactobacillus brevis KL3
Lactobacillus brevis KL5KX 021352 Levilactobacillus brevis KL5
Lactobacillus brevis KL7KX 021354Levilactobacillus brevis KL7
Pediococcus pentosaceus BAL3KX 021370Unchanged name
Pediococcus pentosaceus BAL5KX 021346Unchanged name
Pediococcus pentosaceus BAL6KX 021347Unchanged name
Pediococcus pentosaceus BAL9KX 021349Unchanged name
Pediococcus pentosaceus KL6KX 021353Unchanged name
Pediococcus pentosaceus KL9KX 021361Unchanged name
Pediococcus pentosaceus KL11KX 021362Unchanged name
Pediococcus pentosaceus KL13KX 021363Unchanged name
Pediococcus pentosaceus KL14KX 021364Unchanged name
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Łepecka, A.; Szymański, P.; Rutkowska, S.; Iwanowska, K.; Kołożyn-Krajewska, D. The Influence of Environmental Conditions on the Antagonistic Activity of Lactic Acid Bacteria Isolated from Fermented Meat Products. Foods 2021, 10, 2267. https://doi.org/10.3390/foods10102267

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Łepecka A, Szymański P, Rutkowska S, Iwanowska K, Kołożyn-Krajewska D. The Influence of Environmental Conditions on the Antagonistic Activity of Lactic Acid Bacteria Isolated from Fermented Meat Products. Foods. 2021; 10(10):2267. https://doi.org/10.3390/foods10102267

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Łepecka, Anna, Piotr Szymański, Sylwia Rutkowska, Kinga Iwanowska, and Danuta Kołożyn-Krajewska. 2021. "The Influence of Environmental Conditions on the Antagonistic Activity of Lactic Acid Bacteria Isolated from Fermented Meat Products" Foods 10, no. 10: 2267. https://doi.org/10.3390/foods10102267

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