Next Article in Journal
Structural Characterization of Low-Rank Coals in the Ningdong Coalfield Under the Control of the First Coalification Jump
Previous Article in Journal
Multi-Level Progressive Parameter Optimization Method for the Complex Process Industry
Previous Article in Special Issue
Simulation and Evaluation of Processing Technologies for the Valorization of Sargassum
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Processes
Volume 13
Issue 7
10.3390/pr13071995
processes-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

A Study on Combined Effect of Monoterpenoid Essential Oils and Polyphenols from Grape Pomace on Selected Pathogenic and Probiotic Bacteria Strains

by
Haona Bao
1,2,
Cosmas Mwendwa Muasya
1,
Fur-Chi Chen
1,
Agnes Kilonzo-Nthenge
1 and
Ying Wu
1,*
1
Department of Food and Animal Sciences, Tennessee State University, Nashville, TN 37209, USA
2
Department of Food Science and Technology, The Ohio State University, Columbus, OH 43210, USA
*
Author to whom correspondence should be addressed.
Processes 2025, 13(7), 1995; https://doi.org/10.3390/pr13071995
Submission received: 25 March 2025 / Revised: 13 June 2025 / Accepted: 17 June 2025 / Published: 24 June 2025
(This article belongs to the Special Issue Phytochemicals: Extraction, Optimization, Identification, Biological Activities, and Applications in the Food, Nutraceutical, and Pharmaceutical Industries (2nd Edition))
Browse Figures
Versions Notes

Abstract

In the current study, the combined effects of essential oils (EOs) and polyphenols (PPs) were investigated as potential antibiotic alternatives. Thymol and carvacrol at a ratio of 1:1 was used as the EO due to their well-proven synergistic antimicrobial effect. The PPs were an extracted and freeze-dried product from grape pomace. The treatment solutions were prepared at the EO to PP ratios of 0:10, 3:7, 5:5, 7:3, and 10:0, at total concentrations ranging from 0 to 1000 ppm. The minimum inhibition concentrations were examined on the pathogenic bacteria strains, namely Escherichia coli O157:H7, Salmonella typhimurium, and Enterococcus cloacae, and a probiotic bacterium, Lactiplantibacillus plantarum. The mixed EOs and PPs exhibited varied levels of antibacterial effect against the tested pathogenic bacteria. The MIC of EO and the EO/PP ratio of 10:0 was the best, followed by 7:3 and 5:5 of EO/PP mixed ratios. For the probiotic bacterium, the PP and EO/PP mixed solutions exhibited promoting effects on the growth of L. plantarum at certain concentrations. The results have demonstrated that the combined EOs and PPs could maximize antibacterial activity against pathogenic bacteria while stimulating the growth of the tested probiotic bacterium. This finding will provide useful insights for developing antibiotic alternatives while minimizing the adverse effects on probiotic strains.

1. Introduction

The widespread misuse of antibiotics has led to bacterial resistances of pathogenic strains which bring significant threats to public health, as the residues of antibiotics can directly affect the human immune system, growth, and metabolism processes [1,2]. Each year in the United States, at least 2.8 million people contract antibiotic-resistant infections, and more than 35,000 people die [3]. The regulatory restrictions on using antibiotics in poultry production also induce infectious diseases in poultry such as avian coccidiosis and necrotic enteritis, which result in decreased growth performance, increased mortality, and reduced welfare [4]. Other poultry gastrointestinal infections are caused by Salmonella typhimurium and Escherichia coli, which can survive and pass through the chicken production process and increase the danger of contamination in chicken products, as well as human gastrointestinal infections [5,6]. E. coli O157:H7, S. typhimurium, and Enterococcus cloacae are frequently detected in poultry. They can easily survive and pass through poultry production procedures, thus causing risks in product contamination and human gastrointestinal infections [6]. L. plantarum is an active microbe beneficial to poultry by improving host intestinal microbial balance. L. plantarum can inhibit oocyst shedding and improve the growth performance as well as the intestinal health of broilers infected with Eimeria spp. and Clostridium perfringens which cause infectious diseases [7].
To overcome the problems associated with the ban on using in-feed antibiotics on poultry production as growth promoters, alternatives were sought to combat infectious diseases. Some nutritional feed additives in poultry diets have shown some potential in growth promotion while reducing diseases, these additives include prebiotics, probiotics, bacteriophages, organic acids, essential oils, and natural phytochemical extracts [7].
Among the various natural antibiotic alternatives, phytochemicals have gained increasing attentions in recent years because of their multifaceted antimicrobial mechanisms. Also, bacteria are less likely to develop resistance to them compared with antibiotics [8,9,10]. Essential oils (EOs) are naturally synthesized in different plant organs as secondary metabolites [11]. Thymol and carvacrol are phenolic components and their antibacterial properties are well reported [12]. More studies are needed to fully understand their interactions with pathogenic bacteria, probiotics, and the gut microbiota in humans, livestock, and poultry. For example, thyme oil may inhibit probiotic bacterial strains, including those from the Lactiplantibacillus group [13]. Despite this inhibition of beneficial bacteria, their metabolites can exhibit antibacterial activity against strains of Salmonella enterica [1]. Polyphenols (PPs) are extracted from fruits or plants such as grape pomace, with excellent antioxidant effects, including compounds like flavones, flavanols, anthocyanins, and flavanols [8]. Most studies have concluded that phenolic compounds are effective on bacteria by disrupting cell membranes and inhibiting ATPase activity, etc. [14]. In addition, it is also suggested that flavonoids in PPs may interfere with specific intracellular or surface enzymes and many bacterial virulence factors, such as toxins, enzymes, and signal receptors [15]. EOs and PPs help improve gut histology by increasing villi height, thus improving digestibility and growth performance [8,16,17].
The synergistic interaction implies that the effect of two drugs/chemicals taken together is greater than the sum of their separate effects at the same doses [18]. Although thymol, carvacrol, thymol and carvacrol mixtures, and polyphenols have been investigated as antimicrobial agents previously, the combined effect of EOs and PPs has been scarcely reported. Therefore, in the current study, the combination of EOs (thymol and carvacrol at a ratio of 1:1) and PPs from grape pomace has been investigated to understand their possible synergistic antibacterial activity.

2. Results

According to the results, the five treatments (EO/PP at the ratios of 0:10, 3:7, 5:5, 7:3, and 10:0) were tested for antibacterial activity and showed significant growth inhibition in E. coli O157:H7, S. typhimurium, and E. cloacae. The EO exhibited inhibition in the growth of L. plantarum, while the PP and the EO/PP mixtures showed a growth promoting effect at most tested concentrations. The MICs were recorded as turbidity values obtained spectrophotomically at 600 nm and compared against a blank control (dilution of EO, PP, and their mixtures at corresponding ratios). A negative control was also tested using ethanol to determine whether it had a killing effect on the bacteria; it was observed that the solvent ethanol did not have an effect on the growth of the selected bacteria. There was a significant difference in the inhibition activity between treatments themselves to a particular bacterium and amongst the selected microorganisms. The subcellular microstructures of different bacteria usually vary from each other, and the modes and molecular targets of bacterial inactivation caused by different natural antibacterial agents are also different [19]. Therefore, bioactive compounds often have different antimicrobial spectrums, and their inhibitory effects on the same bacteria may also differ greatly. Furthermore, the chemical composition of EOs and PPs can vary based on the plant species, geographical origin, harvesting period, and extraction methods. This could possibly result in the variation in the susceptibility of Gram-positive and Gram-negative bacteria [20].

2.1. Minimum Inhibition Concentration of Pathogenic Bacteria

As indicated in Figure 1, E. coli O157:H7 showed a similar trend in growth inhibition after being subjected to different treatment solutions and concentrations. At a concentration of 250 ppm, a noticeable inhibition in growth was shown by comparing the OD 600 nm reading from 0.937 for the positive control to the 0.273 and 0.268 for treatments of EO/PP ratios of 7:3 and 5:5, respectively.
The antibacterial effects of the EO/PP mixtures at the ratios of 5:5 and 7:3 on E. coli O157:H7 had no significant difference (p > 0.05). The MIC for EO-only treatment was 500 ppm, while for the EO/PP mixtures at ratios of 5:5 and 7:3, the MICs increased to 1000 ppm, while the MICs of the EO/PP ratio of 3:7 and the PP-only treatments were larger than 1000 ppm (>1000 ppm), as shown in Table 1.
For S. typhimurium, as shown in Figure 2, pure EO and the 5:5 and 7:3 (EO/PP) groups showed a stronger growth inhibition effect, among which the pure EO group showed a significant stronger growth inhibition effect (p < 0.05) compared with the 5:5 and 7:3 EO/PP groups, and had an MIC at 250 ppm, while the 5:5 and 7:3 EO/PP groups showed no difference at all test concentrations, and both had MICs at 500 ppm. The EO/PP ratio of the 3:7 group showed less of an inhibition effect compared with the above three groups but had its MIC at 500 ppm as well. The pure PP treatment showed the lowest growth inhibition effect on S. typhimurium, and the MIC was determined at 1000 ppm. The results indicated that E. coli and S. typhimurium were comparable in their sensitivity to EOs and PPs, whereas EO/PP at ratios of 7:3 and 5:5 had a promising killing effect. Many antimicrobials and EOs that have efficacy against E. coli have also been found to be active against S. typhimurium [21,22,23,24]. For instance, in the study of Severino et al. [23], the MIC tested was 500 ppm for both E. coli and S. typhimurium using citrus oil. Yang et al. [24] also found similar MICs using EO extracts from oregano against E. coli and Salmonella enteritidis.
The inhibition activity of all treatments on E. cloacae are shown in Figure 3. At the lower concentration range, 0–31.25 ppm, there was no significant difference (p > 0.05) in growth amongst all treatments. As the concentration increased to 62.5 ppm, pure EO showed significant inhibition compared with the rest of the groups. In the 125–250 ppm range, pure EO and the 7:3 (EO/PP) mixture had no significant difference in inhibition, suggesting that much of the inhibition effect was due to a higher EO portion in the formula. Similar to the results of E. coli, pure PP did not have obvious effect on bacteria growth within the rested range, but when combined with EO, the MIC decreased.

2.2. Effect on Growth of Lactiplantibacillus Plantarum

The individual EOs and PPs showed opposite effects on the growth of L. plantarum. PP was observed to have a growth enhancing effect on L. plantarum within the tested concentrations, as shown in Figure 4. Specifically, while pure PP had a significant growth enhancement effect, pure EO had an overall inhibition effect on L. plantarum particularly at concentrations above 125 ppm. As shown in Figure 4, the growth of L. plantarum in the control group performed worse than most treatment groups which may be due to the media conditions caused by the treatment compounds, as demonstrated by Horn and Aspmo et al. [25]. Polyphenols in most fruits (such as grapes) are recognized as the major class of phytochemicals with antioxidant activity. Various bacterial species exhibit different sensitivities towards phenolic compounds in the grape. E. coli is very sensitive to wine extracts [26]. Grape seed extracts encourage the antioxidant with the growth of beneficial bacteria and eliminate the growth of pathogenic bacteria in broiler intestines [27].
It is evident that PPs promote the growth of L. plantarum. This observation is consistent with Kim et al. [28], who found that polyphenols in mango (Mangifera indica L.) had significantly increased the abundance of Lactiplantibacillus spp., like L. plantarum and L. reuteri. Axling et al. [29] found that L. plantarum was able to metabolize phenolic acid in green tea, which in turn acted as a probiotic compound to promote gut health. In addition, De Llano et al. [30] reported that flavan-3-ol obtained from grape extracts was observed to promote the growth of Lactiplantibacillus spp. and hinder the growth of Clostridium spp. and Staphylococcus spp.

3. Discussion

The antibacterial activity of EOs against E. coli O157:H7, Salmonella serovars, Listeria monocytogenes, Shigella dysenteriae, Bacillus cereus, and Staphylococcus aureus have been well demonstrated in various in vitro studies as summarized in the review by Burt [31]. These properties may be attributed to their capability to work as antioxidant, antimicrobial, immunomodulatory, and anti-inflammatory agents by suppressing harmful free radicals from interacting with cellular biological compounds and increasing the digestion, absorption, and metabolism of nutrients [32]. A hydrophobic essential oil can partition in the lipophilic interior of a cell membrane and trigger the leakage of cellular contents, resulting in bacterial cell death [33] which alters the gut microbiota by inhibiting the growth of potentially harmful intestinal bacteria in broilers.
The results indicated that all EO and PP mixtures had exhibited inhibitory effects on the tested pathogenic bacteria. EO alone had the highest inhibition effect, and PP alone had the lowest inhibition effect. The inhibitory effects of the EO/PP mixtures at 5:5 and 7:3 were significantly higher than the 3:7 group, where treatments showed inhibition at low concentrations and MICs varied from as low as 500 ppm for S. typhimurium and E. cloacae, and 1000 ppm for E. coli O157:H7. The reported MICs showed discrepancies, such as in the study of Yap et al. [34], where EO extracts from basil and cinnamon had MICs of 1.25 µg/mL and 5 µg/mL against E. coli, respectively. Xue et al. [7] mentioned that when EOs, eugenol and thymol, were used separately, S. enteritidis was less susceptible at 500 ppm. In the current study, the inhibition was detected in the range of 250–1000 ppm, with EO exhibiting the strongest inhibition effect, followed by the 7:3, 5:5, and 3:7 of EO/PP mixtures, while the PP showed the weakest inhibition effect. The results indicated that the major inhibition effect came from the Eos; the higher the portion of EO in the EO/PP mixtures, the better the inhibition on growth of the three tested strains. Among the three tested strains, S. typhimurium was more susceptible to EO, and exhibited the lowest MIC, 250 ppm.
The above phenomenon makes it possible that a combination of EOs and PPs can be used to prohibit the growth of pathogenic bacteria while promoting the growth of probiotic bacteria, which can be applied in moderating gut microbiota. EO alone showed the highest inhibition effect on all strains. PP alone had the lowest inhibition on pathogenic strains but a promotion effect on the probiotic strain. Arumugam et al. [35] reported that plant extracts restrained the pathogenic microbes such as E. coli and S. typhimurium while they stimulated the growth of beneficial bacteria strains like L. plantarum. In the current study, a combination of EOs and PPs had a modulation effect on the antimicrobial activity against Gram-negative bacteria, namely Enterobacter spp., Salmonella spp., and E. coli. A recent study by Cheng et al. [36] found synergistic antimicrobial effects of citrus essential oil and tea polyphenols. The results from the current study indicated synergistic effects in some concentrations and EO/PP mixed ratios, such as at 125 ppm and the 5:5 and 7:3 ratios of EO/PP which showed similar or superior antibacterial effects on E. coli O157:H7 and S. typhimurium, although the MIC values did not reflect the synergistic effect. Within the tested concentration ranging from 0 to 1000 ppm, the MICs of PPs were larger than 1000 ppm for E. coli O157:H7, E. cloacae, and L. plantarum, and MIC was determined to be 1000 ppm for S. typhimurium. The MIC values of polyphenols extracted from different plants varied largely, e.g., from a few ppm to 2000 ppm [37], which may be attributed to the chemical composition, purity, and selected bacteria strains.

4. Materials and Methods

4.1. Essential Oils and Polyphenols Stock Solution

The grape pomace extract (exGrape®Total) was purchased from Lalilab Inc. (Durham, NC, USA). The exGrape®Total is a unique profile of polyphenols from red grape pomace extracted using water extraction, polyphenol purification and standardization, and spray drying [38]. All chemicals, thymol (≥98.5%), carvacrol (≥99%), reagents, and materials were purchased from SigmaAldrich (St. Louis, MO, USA).
EOs (thymol and carvacrol at the ratio of 1:1) and PPs from grape pomace extracts were dissolved and vortexed thoroughly in 70% ethanol to make a final concentration of 5% (w/w) each to make treatment inoculums. Five treatments with different EO and PP ratios ranging from 0:10, 3:7, 5:5, 7:3, and 10:0 were made from the stock solutions.

4.2. Bacteria Culture

E. coli O157:H7, S. typhimurium, E. cloacae, and L. plantarum were used as testing species as shown in Table 1. Overnight cultures (incubated at 37 °C) were diluted with Mueller–Hinton Broth (MHB) yielding a suspension close to 107 CFU/mL to make the control inoculum. The control inoculum (240 µL) was dispensed into 96-well micro plates using clean MHB as a negative control. Antimicrobial treatment solution (10 µL) was dispensed to the wells starting with the controls and the least concentrated inoculums. Negative controls contained no bacteria (removing background color effect of the compounds), and the positive controls had bacteria but no treatment solutions. A solvent control containing an equilibrant of EtOH of each dilution was also placed in the well. The plates were incubated at 37 °C for 24 h aerobically for E. coli O157:H7, S. typhimurium, and E. cloacae, and anaerobically for L. plantarum using an anaerobic jar. The turbidity was measured at 600 nm using a microplate reader (Benchmark Plus, Bio-Rad, CA, USA). Experiments were performed in duplicate with three repeats.

4.3. Antibacterial Activity

The antibacterial activity of EOs, PPs, and their combinations was determined by minimum inhibitory concentration by the micro dilution assay in 96-well plates according to Man, A. et al. with modifications [39]. MICs were expressed as a minimum concentration, which prevented the visible growth (≤0.05 difference in OD600) in the wells. For each ratio, the antibacterial activity was tested against three Gram-negative bacteria and a probiotic Gram-positive bacterium (Table 2). Each ratio (antimicrobial agent) was micro diluted using Mueller–Hinton Broth (MBH) (St. Louis, MO, USA). and PP solutions were mixed at various ratios (0:10, 3:7, 5:5, 7:3, and 10:0) and added to MHB to make 1000 ppm solutions. Two-fold dilutions were conducted to make 500.00, 250.00, 125.00, 62.50, 31.25, 15.63, and 7.81 ppm solutions, with 0 ppm (no test compound) used as a control. Blanks for each dilution of EO, PP, and their mixed solutions were tested for OD reading.
Chen et al. [37] have summarized the antimicrobial mechanism of polyphenols including changing the membrane function, disturbing intracellular function, and apoptosis-like death. They have also summarized that polyphenols can inhibit the growth of pathogenic bacteria, while promoting the growth of beneficial probiotic bacteria, which agrees with the results of the current study.

4.4. Statistical Analysis

All measurements were tested in triplicate, and the data were presented as mean ± standard deviation. Statistical analyses were conducted using the Statistical Analysis System (SAS Release 9.3, SAS Institute Inc., Cary, NC, USA). One-way analysis of variance and Tukey’s HSD tests were used to test the significance of difference among samples at a level p < 0.05.

5. Conclusions

There is limited research on the effects of combined natural extracts on the growth of both pathogenic and probiotic bacterial strains. In the current work, it was observed that all the pathogenic bacteria strains were inhibited by the EO and PP combinations at varied levels. At certain concentrations (31.5–250 ppm), the mixed EOs and PPs at ratios of 5:5 and 7:3 not only prohibited the growth of pathogenic bacterial strains, but also promoted the growth of the beneficial bacterium, L. plantarum. Synergistic antibacterial effects were observed at 125 ppm and in the 5:5 and 7:3 EO/PP groups on E. coli O157:H7 and S. typhimurium. The findings could be further utilized in developing antibiotic alternatives to prohibit pathogenic bacteria strains and to promote the growth of probiotic bacteria, thus modulating the gut microbiome profile in livestock and poultry.

Author Contributions

Conceptualization, Y.W.; methodology, F.-C.C. and A.K.-N.; validation, Y.W., F.-C.C., and A.K.-N.; formal analysis, H.B. and C.M.M.; investigation, C.M.M.; resources, Y.W.; data curation, H.B. and Y.W.; writing—original draft preparation, C.M.M., H.B., and Y.W.; writing—review and editing, Y.W.; visualization, Y.W.; supervision, Y.W., F.-C.C., and A.K.-N.; project administration, Y.W.; funding acquisition, Y.W. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by the Capacity Building Program, project award no. 2019-38821-29055, from the U.S. Department of Agriculture’s National Institute of Food and Agriculture.

Data Availability Statement

The raw data supporting the conclusions of this article will be made available by the authors on request.

Acknowledgments

The authors would like to thank Yvonne G. Myles for her technical support.

Conflicts of Interest

The authors declare no conflicts of interest.

Abbreviations

The following abbreviations are used in this manuscript:
EOEssential Oil
MICMinimal Inhibition Concentration
PPPolyphenol

References

  1. Bartkiene, E.; Ruzauskas, M.; Bartkevics, V.; Pugajeva, I.; Zavistanaviciute, P.; Starkute, V.; Zokaityte, E.; Lele, V.; Dauksiene, A.; Grashorn, M.; et al. Study of the Antibiotic Residues in Poultry Meat in Some of the EU Countries and Selection of the Best Compositions of Lactic Acid Bacteria and Essential Oils against Salmonella enterica. Poult. Sci. 2020, 99, 4065–4076. [Google Scholar] [CrossRef] [PubMed]
  2. Kaur, K.; Singh, S.; Kaur, R. Impact of Antibiotic Usage in Food-Producing Animals on Food Safety and Possible Antibiotic Alternatives. Microbe 2024, 4, 100097. [Google Scholar] [CrossRef]
  3. Center for Disease Control and Prevention. Antibiotic Resistance Threats in the United States; CDC: Atlanta, GA, USA, 2020.
  4. Adhikari, P.; Kiess, A.; Adhikari, R.; Jha, R. An Approach to Alternative Strategies to Control Avian Coccidiosis and Necrotic Enteritis. J. Appl. Poult. Res. 2020, 29, 515–534. [Google Scholar] [CrossRef]
  5. Projahn, M.; Pacholewicz, E.; Becker, E.; Correia-Carreira, G.; Bandick, N.; Kaesbohrer, A. Reviewing Interventions against Enterobacteriaceae in Broiler Processing: Using Old Techniques for Meeting the New Challenges of ESBL E. coli? BioMed Res. Int. 2018, 2018, 7309346. [Google Scholar] [CrossRef]
  6. Vinueza-Burgos, C.; Baquero, M.; Medina, J.; De Zutter, L. Occurrence, Genotypes and Antimicrobial Susceptibility of Salmonella Collected from the Broiler Production Chain within an Integrated Poultry Company. Int. J. Food Microbiol. 2019, 299, 1–7. [Google Scholar] [CrossRef]
  7. Xue, F.; Shi, L.; Li, Y.; Ni, A.; Ma, H.; Sun, Y.; Chen, J. Effects of Replacing Dietary Aureomycin with a Combination of Plant Essential Oils on Production Performance and Gastrointestinal Health of Broilers. Poult. Sci. 2020, 99, 4521–4529. [Google Scholar] [CrossRef]
  8. Balta, I.; Linton, M.; Pinkerton, L.; Kelly, C.; Stef, L.; Pet, I.; Stef, D.; Criste, A.; Gundogdu, O.; Corcionivoschi, N. The Effect of Natural Antimicrobials against Campylobacter spp. and Its Similarities to Salmonella spp., Listeria spp., Escherichia coli, Vibrio spp., Clostridium spp. and Staphylococcus spp. Food Control 2021, 121, 107745. [Google Scholar] [CrossRef]
  9. Marchese, A.; Orhan, I.E.; Daglia, M.; Barbieri, R.; Di Lorenzo, A.; Nabavi, S.F.; Gortzi, O.; Izadi, M.; Nabavi, S.M. Antibacterial and Antifungal Activities of Thymol: A Brief Review of the Literature. Food Chem. 2016, 210, 402–414. [Google Scholar] [CrossRef]
  10. Radünz, M.; dos Santos Hackbart, H.C.; Camargo, T.M.; Nunes, C.F.P.; de Barros, F.A.P.; Dal Magro, J.; Filho, P.J.S.; Gandra, E.A.; Radünz, A.L.; da Rosa Zavareze, E. Antimicrobial Potential of Spray Drying Encapsulated Thyme (Thymus vulgaris) Essential Oil on the Conservation of Hamburger-like Meat Products. Int. J. Food Microbiol. 2020, 330, 108696. [Google Scholar] [CrossRef]
  11. El Asbahani, A.; Miladi, K.; Badri, W.; Sala, M.; Addi, E.H.A.; Casabianca, H.; El Mousadik, A.; Hartmann, D.; Jilale, A.; Renaud, F.N.R.; et al. Essential Oils: From Extraction to Encapsulation. Int. J. Pharm. 2015, 483, 220–243. [Google Scholar] [CrossRef]
  12. Bao, H.; Ding, H.H.; Charles, A.P.R.; Hui, D.; Rakshit, S.; Nahashon, S.; Wu, Y. Application of Yellow Mustard Mucilage in Encapsulation of Essential Oils and Polyphenols Using Spray Drying. Food Hydrocoll. 2023, 143, 108815. [Google Scholar] [CrossRef]
  13. de Souza Rodrigues, H.L.; Kolososki, I.M.M.; Benevides, V.P.; Saraiva, M.M.S.; Berchieri Junior, A. Essential Oils Used in the Poultry Industry: Would It Be an Effective Green Alternative against Salmonella spp. Dissemination and Antimicrobial Resistance? Microbe 2025, 6, 100248. [Google Scholar] [CrossRef]
  14. Valcourt, C.; Saulnier, P.; Umerska, A.; Zanelli, M.P.; Montagu, A.; Rossines, E.; Joly-Guillou, M.L. Synergistic Interactions between Doxycycline and Terpenic Components of Essential Oils Encapsulated within Lipid Nanocapsules against Gram Negative Bacteria. Int. J. Pharm. 2016, 498, 23–31. [Google Scholar] [CrossRef] [PubMed]
  15. Kaschubek, T.; Mayer, E.; Rzesnik, S.; Grenier, B.; Bachinger, D.; Schieder, C.; König, J.; Teichmann, K. Effects of Phytogenic Feed Additives on Cellular Oxidative Stress and Inflammatory Reactions in Intestinal Porcine Epithelial Cells1. J. Anim. Sci. 2018, 96, 3657–3669. [Google Scholar] [CrossRef]
  16. Zumbaugh, C.A.; Murugesan, G.R.; Wong, E.A.; Syed, B.; Persia, M.E. Evaluation of a Phytogenic Feed Additive on Performance, Nutrient Digestion, and Absorption in Turkey Poults. Anim. Feed Sci. Technol. 2020, 267, 114575. [Google Scholar] [CrossRef]
  17. Gadde, U.; Kim, W.H.; Oh, S.T.; Lillehoj, H.S. Alternatives to Antibiotics for Maximizing Growth Performance and Feed Efficiency in Poultry: A Review. Anim. Health Res. Rev. 2017, 18, 26–45. [Google Scholar] [CrossRef]
  18. Pei, R.; Zhou, F.; Ji, B.; Xu, J. Evaluation of Combined Antibacterial Effects of Eugenol, Cinnamaldehyde, Thymol, and Carvacrol against E. Coli with an Improved Method. J. Food Sci. 2009, 74, M379–M383. [Google Scholar] [CrossRef]
  19. Rai, M.; Paralikar, P.; Jogee, P.; Agarkar, G.; Ingle, A.P.; Derita, M.; Zacchino, S. Synergistic Antimicrobial Potential of Essential Oils in Combination with Nanoparticles: Emerging Trends and Future Perspectives. Int. J. Pharm. 2017, 519, 67–78. [Google Scholar] [CrossRef]
  20. Bassanetti, I.; Carcelli, M.; Buschini, A.; Montalbano, S.; Leonardi, G.; Pelagatti, P.; Tosi, G.; Massi, P.; Fiorentini, L.; Rogolino, D. Investigation of Antibacterial Activity of New Classes of Essential Oils Derivatives. Food Control. 2017, 73, 606–612. [Google Scholar] [CrossRef]
  21. Duarte, I.F.; Barros, A.; Almeida, C.; Spraul, M.; Gil, A.M. Multivariate Analysis of NMR and FTIR Data as a Potential Tool for the Quality Control of Beer. J. Agric. Food Chem. 2004, 52, 1031–1038. [Google Scholar] [CrossRef]
  22. Olszewska, M.A.; Gędas, A.; Simões, M. Antimicrobial Polyphenol-Rich Extracts: Applications and Limitations in the Food Industry. Food Res. Int. 2020, 134, 109214. [Google Scholar] [CrossRef] [PubMed]
  23. Severino, R.; Ferrari, G.; Vu, K.D.; Donsì, F.; Salmieri, S.; Lacroix, M. Antimicrobial Effects of Modified Chitosan Based Coating Containing Nanoemulsion of Essential Oils, Modified Atmosphere Packaging and Gamma Irradiation against Escherichia coli O157:H7 and Salmonella typhimurium on Green Beans. Food Control. 2015, 50, 215–222. [Google Scholar] [CrossRef]
  24. Yang, C.; Chowdhury, M.A.; Huo, Y.; Gong, J. Phytogenic Compounds as Alternatives to In-Feed Antibiotics: Potentials and Challenges in Application. Pathogens 2015, 4, 137–156. [Google Scholar] [CrossRef]
  25. Horn, S.J.; Aspmo, S.I.; Eijsink, V.G.H. Growth of Lactobacillus Plantarum in Media Containing Hydrolysates of Fish Viscera. J. Appl. Microbiol. 2005, 99, 1082–1089. [Google Scholar] [CrossRef]
  26. Xia, E.-Q.; Deng, G.-F.; Guo, Y.-J.; Li, H.-B. Biological Activities of Polyphenols from Grapes. Int. J. Mol. Sci. 2010, 11, 622–646. [Google Scholar] [CrossRef]
  27. Aditya, S.; Ohh, S.-J.; Ahammed, M.; Lohakare, J. Supplementation of Grape Pomace (Vitis vinifera) in Broiler Diets and Its Effect on Growth Performance, Apparent Total Tract Digestibility of Nutrients, Blood Profile, and Meat Quality. Anim. Nutr. 2018, 4, 210–214. [Google Scholar] [CrossRef]
  28. Kim, H.; Venancio, V.P.; Fang, C.; Dupont, A.W.; Talcott, S.T.; Mertens-Talcott, S.U. Mango (Mangifera indica L.) Polyphenols Reduce IL-8, GRO, and GM-SCF Plasma Levels and Increase Lactobacillus Species in a Pilot Study in Patients with Inflammatory Bowel Disease. Nutr. Res. 2020, 75, 85–94. [Google Scholar] [CrossRef]
  29. Axling, U.; Olsson, C.; Xu, J.; Fernandez, C.; Larsson, S.; Ström, K.; Ahrné, S.; Holm, C.; Molin, G.; Berger, K. Green Tea Powder and Lactobacillus Plantarum Affect Gut Microbiota, Lipid Metabolism and Inflammation in High-Fat Fed C57BL/6J Mice. Nutr. Metab. 2012, 9, 105. [Google Scholar] [CrossRef]
  30. De Llano, D.; Gil-Sánchez, I.; Esteban-Fernández, A.; Ramos, A.; Cueva, C.; Moreno-Arribas, M.; Bartolomé, B. Some Contributions to the Study of Oenological Lactic Acid Bacteria through Their Interaction with Polyphenols. Beverages 2016, 2, 27. [Google Scholar] [CrossRef]
  31. Burt, S. Essential Oils: Their Antibacterial Properties and Potential Applications in Foods—A Review. Int. J. Food Microbiol. 2004, 94, 223–253. [Google Scholar] [CrossRef]
  32. Mohamed, H.G.; Gaafar, A.M.; Soliman, A.S. Antimicrobial Activities of Essential Oil of Eight Plant Species from Different Families Against Some Pathogenic Microorganisms. Res. J. Microbiol. 2016, 11, 28–34. [Google Scholar] [CrossRef]
  33. Moon, S.H.; Waite-Cusic, J.; Huang, E. Control of Salmonella in Chicken Meat Using a Combination of a Commercial Bacteriophage and Plant-Based Essential Oils. Food Control 2020, 110, 106984. [Google Scholar] [CrossRef]
  34. Yap, P.S.X.; Yiap, B.C.; Ping, H.C.; Lim, S.H.E. Essential Oils, A New Horizon in Combating Bacterial Antibiotic Resistance. Open Microbiol. J. 2014, 8, 6–14. [Google Scholar] [CrossRef] [PubMed]
  35. Arumugam, G.; Swamy, M.; Sinniah, U. Plectranthus amboinicus (Lour.) Spreng: Botanical, Phytochemical, Pharmacological and Nutritional Significance. Molecules 2016, 21, 369. [Google Scholar] [CrossRef] [PubMed]
  36. Cheng, F.; Chen, M.; Duan, Z.; Zou, Y.; He, Y.; Zeng, F.; Yuan, Y.; Fu, T.; Tu, H.; Li, R.; et al. Fabrication, Characterization, and Bioactivity of Self-Assembled Carrier-Free Colloidal Dispersions from Citrus × Limon ‘Rosso’ Essential Oil and Tea Polyphenols. Food Chem. 2024, 457, 140058. [Google Scholar] [CrossRef]
  37. Chen, X.; Lan, W.; Xie, J. Natural Phenolic Compounds: Antimicrobial Properties, Antimicrobial Mechanisms, and Potential Utilization in the Preservation of Aquatic Products. Food Chem. 2024, 440, 138198. [Google Scholar] [CrossRef]
  38. UL Prospector, ExGrapeTM Total Product Detail. Available online: https://www.ulprospector.com/en/na/Food/Detail/5677/202864/exGrape-Total (accessed on 18 May 2025).
  39. Man, A.; Santacroce, L.; Iacob, R.; Mare, A.; Man, L. Antimicrobial Activity of Six Essential Oils Against a Group of Human Pathogens: A Comparative Study. Pathogens 2019, 8, 15. [Google Scholar] [CrossRef]
Processes 13 01995 g001
Figure 1. Effect of EO, PP, and EO/PP mixtures on the growth of Escherichia coli 0157:H7. Different letters indicated significant difference (p < 0.05), upper case letters denote the comparison among concentrations within the same treatment group, lower case letters denote the comparisons among different EO/PP ratios at the same concentration.
Figure 1. Effect of EO, PP, and EO/PP mixtures on the growth of Escherichia coli 0157:H7. Different letters indicated significant difference (p < 0.05), upper case letters denote the comparison among concentrations within the same treatment group, lower case letters denote the comparisons among different EO/PP ratios at the same concentration.
Processes 13 01995 g001
Processes 13 01995 g002
Figure 2. Effect of EO, PP, and EO/PP mixtures on the growth of Salmonella typhimurium. Different letters indicate significant difference (p < 0.05), upper case letters denote the comparison among concentrations within the same treatment group, lower case letters denote the comparisons among different EO/PP ratios at the same concentration.
Figure 2. Effect of EO, PP, and EO/PP mixtures on the growth of Salmonella typhimurium. Different letters indicate significant difference (p < 0.05), upper case letters denote the comparison among concentrations within the same treatment group, lower case letters denote the comparisons among different EO/PP ratios at the same concentration.
Processes 13 01995 g002
Processes 13 01995 g003
Figure 3. Effect of EO, PP, and EO/PP mixtures on the growth of Enterococcus cloacae. Different letters indicate significant difference (p < 0.05), upper case letters denote the comparison among concentrations within the same treatment group, lower case letters denote the comparisons among different EO/PP ratios at the same concentration.
Figure 3. Effect of EO, PP, and EO/PP mixtures on the growth of Enterococcus cloacae. Different letters indicate significant difference (p < 0.05), upper case letters denote the comparison among concentrations within the same treatment group, lower case letters denote the comparisons among different EO/PP ratios at the same concentration.
Processes 13 01995 g003
Processes 13 01995 g004
Figure 4. Effect of EO, PP, and EO/PP mixtures on the growth of Lactiplantibacillus plantarum. Different letters indicate significant difference (p < 0.05), upper case letters denote the comparison among concentrations within the same treatment group, lower case letters denote the comparisons among different EO/PP ratios at the same concentration.
Figure 4. Effect of EO, PP, and EO/PP mixtures on the growth of Lactiplantibacillus plantarum. Different letters indicate significant difference (p < 0.05), upper case letters denote the comparison among concentrations within the same treatment group, lower case letters denote the comparisons among different EO/PP ratios at the same concentration.
Processes 13 01995 g004
Table 1. Minimum inhibition concentrations [2] of EO and PP mixtures on tested bacteria.
Table 1. Minimum inhibition concentrations [2] of EO and PP mixtures on tested bacteria.
* EO/PP RatioEscherichia coli O157:H7Salmonella typhimuriumEnterococcus cloacaeLactiplantibacillus plantarum
0/10 (polyphenols only)>1000 ppm1000 ppm>1000 ppm>1000 ppm
3/7>1000 ppm500 ppm1000 ppm>1000 ppm
5/51000 ppm500 ppm1000 ppm>1000 ppm
7/31000 ppm500 ppm500 ppm>1000 ppm
10/0 (essential oils only)500 ppm250 ppm500 ppm500 ppm
* EO/PP: the ratio between essential oils and polyphenols.
Table 2. Bacteria strains used in this study and their culture conditions.
Table 2. Bacteria strains used in this study and their culture conditions.
FamilyGenusSpeciesATCCGrowth ConditionsMedia 1
EnterobacteriaceaeSalmonellatyphimurium23564Aerobic, 18–24 h, 37 °CTSA, MHB
EnterobacteriaceaeEnterococcuscloacae23355Aerobic, 18–24 h, 37 °CTSA, MHB
EnterobacteriaceaeEscherichiacoli25922Aerobic, 18–24 h, 37 °CTSA, MHB
LactobacillaceaeLactiplantibacillusplantarum8014Anaerobic, 48–72 h, 37 °CTSA, MHB
1 TSA: Trypton Soy Agar; MHB: Mueller–Hinton Broth.
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content.

Share and Cite

MDPI and ACS Style

Bao, H.; Muasya, C.M.; Chen, F.-C.; Kilonzo-Nthenge, A.; Wu, Y. A Study on Combined Effect of Monoterpenoid Essential Oils and Polyphenols from Grape Pomace on Selected Pathogenic and Probiotic Bacteria Strains. Processes 2025, 13, 1995. https://doi.org/10.3390/pr13071995

AMA Style

Bao H, Muasya CM, Chen F-C, Kilonzo-Nthenge A, Wu Y. A Study on Combined Effect of Monoterpenoid Essential Oils and Polyphenols from Grape Pomace on Selected Pathogenic and Probiotic Bacteria Strains. Processes. 2025; 13(7):1995. https://doi.org/10.3390/pr13071995

Chicago/Turabian Style

Bao, Haona, Cosmas Mwendwa Muasya, Fur-Chi Chen, Agnes Kilonzo-Nthenge, and Ying Wu. 2025. "A Study on Combined Effect of Monoterpenoid Essential Oils and Polyphenols from Grape Pomace on Selected Pathogenic and Probiotic Bacteria Strains" Processes 13, no. 7: 1995. https://doi.org/10.3390/pr13071995

APA Style

Bao, H., Muasya, C. M., Chen, F.-C., Kilonzo-Nthenge, A., & Wu, Y. (2025). A Study on Combined Effect of Monoterpenoid Essential Oils and Polyphenols from Grape Pomace on Selected Pathogenic and Probiotic Bacteria Strains. Processes, 13(7), 1995. https://doi.org/10.3390/pr13071995

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop