In vitro evaluation of antibacterial and antifungal effects of selected acidifiers and antibacterial effects of essential oils used in poultry feed industry

Document Type : Full paper (Original article)

Authors

1 Department of Animal Health Management, School of Veterinary Medicine, Shiraz University, Shiraz, Iran

2 Graduated from School of Veterinary Medicine, Shiraz University, Shiraz, Iran

3 Ph.D. Student in Feed Hygiene, Department of Animal Health Management, School of Veterinary Medicine, Shiraz University, Shiraz, Iran

Abstract

Background: Essential oils and organic acidifiers are used as natural antimicrobial substitutes for chemical additives in poultry feed. Pathogens such as Salmonella and Escherichia coli pose significant risks. While essential oils possess strong antibacterial and antifungal properties, commercial acidifiers (blends of formic, propionic, and lactic acids) are also commonly applied to reduce microbial contamination in feed. However, comparative data on their efficacy against key poultry pathogens remain limited. Aims: ‏The present study investigates the antibacterial and antifungal effects of selected commonly used commercial acidifiers in poultry feed and also compares them with antibacterial effects of essential oils. Methods: Antibacterial effects against Salmonella typhimurium and E. coli were evaluated for both five commercial acidifiers and essential oils (cinnamon, thyme, clove). Antifungal effects against Fusarium oxysporum were evaluated for acidifiers only. Microplate and agar-well diffusion methods were used. Acidifiers were tested at 1×, 2×, and 5× the manufacturer’s recommended dose; essential oils at 2% and 5% (v/v). Sampling time points for feed pH and microbial counts were days 0, 7, and 21. All tests were performed in triplicate and repeated twice. The impact of acidifiers on feed pH was also evaluated. Results: Acidifiers exhibited negligible inhibitory effects against target pathogens even at concentrations five times higher than recommended levels. In contrast, essential oils demonstrated significant inhibitory effects. Furthermore, the addition of acidifiers did not affect the pH of the feed. Conclusions: The study underscores the potent antibacterial properties of cinnamon, thyme, and clove essential oils against common poultry pathogens. Comparatively, commercial acidifiers showed limited efficacy. Incorporating essential oils into poultry feed can enhance animal health and safety as effective substitutes for chemical additives. Further research is needed to optimize their use and maximize their antimicrobial benefits.

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References

Abd El-Hack, ME; El-Saadony, MT; Saad, AM; Salem, HM; Ashry, NM; Ghanima, MMA; Shukry, M; Swelum, AA; Taha, AE and El-Tahan, AM (2022). Essential oils and their nanoemulsions as green alternatives to antibiotics in poultry nutrition: a comprehensive review. Poult. Sci., 101: 101584. doi: 10.1016/j.psj.2021.101584.
Aguiar Campolina, G; D as Graças Cardoso, M; Rodrigues-Silva-Caetano, A; Lee Nelson, D and Mendes Ramos, E (2023). Essential oil and plant extracts as preservatives and natural antioxidants applied to meat and meat products: a review. Food Technol. Biotechnol., 61: 212-225. doi: 10.17113/ftb.61.02.23.7883.
Aliverdi-Nasab, K; Zhandi, M; Yousefi, AR; Zahedi, V and Rafieian-Naeini, HR (2023). The effect of acidifier supplementation on egg production performance and intestinal histology of Japanese quail (Coturnix japonica). Vet. Med. Sci., 9: 263-271. doi: 10.1002/vms3.1043.
Andrade-Ochoa, S; Chacón-Vargas, KF; Sánchez-Torres, LE; Rivera-Chavira, BE; Nogueda-Torres, B and Nevárez-Moorillón, GV (2021). Differential antimicrobial effect of essential oils and their main components: insights based on the cell membrane and external structure. Membranes. 11: 405. doi: 10.3390/membranes11060405.
AOAC (2005). Official methods of analysis (18th Edn.), Association of Official Analytical Chemists, Gaithersburg, MD.
Bernardini, L; Barbosa, E; Charão, MF and Brucker, N (2022). Formaldehyde toxicity reports from in vitro and in vivo studies: a review and updated data. Drug Chem. Toxicol., 45: 972-984. doi: 10.1080/01480545.2020. 1795190.
Brown, KJ; Tannock, GW; Elliott, RB andLines, DR (1980). Comparison of liquid and agar-solidified defined media regarding the physiological mechanism by which β-2-thienylalanine inhibits growth of Escherichia, Shigella, and Salmonella cultures. Microbiol. Immunol., 24: 603-615. doi: 10.1111/j.1348-0421.1980.tb00664.x.
Burt, S (2004). Essential oils: their antibacterial properties and potential applications in foods—a review. Int. J. Food Microbiol., 94: 223-253. doi: 10.1016/j.ijfoodmicro.2004.03. 022.
Casalino, G; Dinardo, FR; D’Amico, F; Bozzo, G; Bove, A; Camarda, A; Lombardi, R; Dimuccio, MM and Circella, E (2023). Antimicrobial efficacy of cinnamon essential oil against avian pathogenic Escherichia coli from poultry. Animals. 13: 2639. doi: 10.3390/ani13162639.
Chaieb, K; Hajlaoui, H; Zmantar, T; Kahla-Nakbi, AB; Rouabhia, M; Mahdouani, K and Bakhrouf, A (2007). The chemical composition and biological activity of clove essential oil, Eugenia caryophyllata (Syzigium aromaticum L. Myrtaceae): a short review. Phytother. Res., 21: 501-506. doi: 10.1002/ptr.2124.
Chen, L; Zhao, X; Li, R and Yang, H (2022). Integrated metabolomics and transcriptomics reveal the adaptive responses of Salmonella enterica serovar Typhimurium to thyme and cinnamon oils. Food Res. Int., 157: 111241. doi: 10.1016/j.foodres.2022.111241.
Choroszy-Król, I; Futoma-Kołoch, B; Kuźnik, K; Wojnicz, D; Tichaczek-Goska, D; Frej-Mądrzak, M; Jama-Kmiecik, A and Sarowska, J (2023). Exposing Salmonella Senftenberg and Escherichia coli strains isolated from poultry farms to formaldehyde and lingonberry extract at low concentrations. Int. J. Mol. Sci., 24: 14579. doi: 10.3390/ijms241914579.
Eslami, N; Anzabi, Y and Nourazar, MA (2022). The role of Thymus vulgaris essential oil on the rate of fecal excretion of Salmonella typhimurium in broilers infected with bacteria. J. Zoonotic Dis., 6: 157-167. doi: 10.22034/jzd. 2022.15683.
Evangelista, AG; Corrêa, JAF; Pinto, ACSM and Luciano, FB (2022). The impact of essential oils on antibiotic use in animal production regarding antimicrobial resistance – a review. Crit. Rev. Food Sci. Nutr., 62: 5267-5283. doi: 10.1080/10408398. 2021.1883548.
Farouk, MM; El-Molla, A; Salib, FA; Soliman, YA and Shaalan, M (2020). The role of silver nanoparticles in a treatment approach for multidrug-resistant Salmonella species isolates. Int. J. Nanomed., 15: 6993-7011. doi: 10.2147/IJN.S270204.
Friedman, M (2017). Chemistry, antimicrobial mechanisms, and antibiotic activities of cinnamaldehyde against pathogenic bacteria in animal feeds and human foods. J. Agric. Food Chem., 65: 10406-10423. doi: 10.1021/acs. jafc.7b04344.
Gosling, RJ; Mawhinney, I; Richardson, K; Wales, A and Davies, R (2021). Control of Salmonella and pathogenic E. coli contamination of animal feed using alternatives to formaldehyde-based treatments. Microorganisms. 9: 263. doi: 10.3390/microorganisms9020263.
Gupta, C; Garg, AP; Uniyal, RC and Kumari, A (2008). Comparative analysis of the antimicrobial activity of cinnamon oil and cinnamon extract on some food-borne microbes. Afr. J. Microbiol. Res., 2: 247-251.
Hąc-Wydro, K; Flasiński, M and Romańczuk, K (2017). Essential oils as food eco-preservatives: model system studies on the effect of temperature on limonene antibacterial activity. Food Chem., 235: 127-135. doi: 10.1016/j.foodchem.2017.05.051.
Irawan, A; Hidayat, C; Jayanegara, A and Ratriyanto, A (2021). Essential oils as growth-promoting additives on performance, nutrient digestibility, cecal microbes, and serum metabolites of broiler chickens: a meta-analysis. Anim. Biosci., 34: 1499-1513. doi.: 10.5713/ab.20.0668.
Khwaza, V and Aderibigbe, BA (2025). Antibacterial activity of selected essential oil components and their derivatives: a review. Antibiotics. 14: 68. doi: 10.3390/antibiotics14010068.
Kissels, W; Wu, X and Santos, R (2017). Interaction of the isomers carvacrol and thymol with the antibiotics doxycycline and tilmicosin: in vitro effects against pathogenic bacteria commonly found in the respiratory tract of calves. J. Dairy Sci., 100: 970-974. doi: 10.3168/ jds.2016-11536.
Liu, T; Kang, J and Liu, L (2021). Thymol as a critical component of Thymus vulgaris L. essential oil combats Pseudomonas aeruginosa by intercalating DNA and inactivating biofilm. LWT. 136: 110354. doi: 10.1016/j.lwt. 2020.110354.
Loan, NVT; Thong, HT and Duc, HV (2023). Acidifiers as alternatives for antibiotics reduction and gut health improvement for poultry and swine. In: Babinszky, L; Oliveira, J and Santos, EM (Eds.), Feed additives - recent trends animal nutrition. IntechOpen London, UK. PP: 1-22. doi: 10.5772/intechopen.112963.
Magi, G; Marini, E and Facinelli, B (2015). Antimicrobial activity of essential oils and carvacrol, and synergy of carvacrol and erythromycin, against clinical, erythromycin-resistant Group A Streptococci. Front. Microbiol., 6: 165. doi: 10.3389/fmicb.2015.00165.
Mantzios, T; Tsiouris, V; Kiskinis, K; Economou, V; Petridou, E; Tsitsos, A; Patsias, A; Apostolou, I; Papadopoulos, GA and Giannenas, I (2023). In vitro investigation of the antibacterial activity of nine commercial water disinfectants, acidifiers, and glyceride blends against the most important poultry zoonotic bacteria. Pathogens. 12: 381. doi: 10.3390/pathogens12030381.
Mincheva, N; Oblakova, M; Ivanova, I and Hristakieva, P (2025). Economic evaluation of broiler diets supplemented with either selected herbs or their associated essential oils. Black Sea J. Agric., 8: 3-4. doi: 10.47115/bsagriculture. 1564174.
Morshdy, AEM; El-Tahlawy, AS; Qari, SH; Qumsani, AT; Bay, DH; Sami, R; Althubaiti, EH; Mansour, AM; Aljahani, AH and Hafez, AE-SE (2022). Anti-biofilms’ activity of garlic and thyme essential oils against Salmonella typhimurium. Molecules. 27: 2182. doi: 10.3390/molecules27072182.
Movahedi, F; Nirmal, N; Wang, P; Jin, H; Grøndahl, L and Li, L (2024). Recent advances in essential oils and their nanoformulations for poultry feed. J. Anim. Sci. Biotechnol., 15: 110. doi: 10.1186/s40104-024-01067-8.
Mucha, W and Witkowska, D (2021). The applicability of essential oils in different stages of production of animal-based foods. Molecules. 26: 3798. doi: 10.3390/molecules26133798.
Nehme, R; Andrés, S; Pereira, RB; Ben Jemaa, M; Bouhallab, S; Ceciliani, F; López, S; Rahali, FZ; Ksouri, R and Pereira, DM (2021). Essential oils in livestock: from health to food quality. Antioxidants. 10: 330. doi: 10.3390/antiox10020330.
Nguyen, DH; Seok, WJ and Kim, IH (2020). Organic acids mixture as a dietary additive for pigs—a review. Animals. 10: 952. doi: 10.3390/ani10060952.
Omidi, A; Mosleh, H and Rahimi-Kakolaki, M (2024). Comparative evaluation of disinfectants’ efficacy in reducing bacterial and fungal contamination in livestock feed production. Iran. J. Vet. Med., 19: 385-396. doi: 10.32598/ijvm.19.2.1005480.
Pearlin, BV; Muthuvel, S; Govidasamy, P; Villavan, M; Alagawany, M; Ragab Farag, M; Dhama, K and Gopi, M (2020). Role of acidifiers in livestock nutrition and health: a review. J. Anim. Physiol. Anim. Nutr., 104: 558-569. doi: 10.1111/jpn.13232.
Polycarpo, GV; Andretta, I; Kipper, M; Cruz-Polycarpo, VC; Dadalt, JC; Rodrigues, PHM and Albuquerque, RD (2017). Meta-analytic study of organic acids as an alternative performance-enhancing feed additive to antibiotics for broiler chickens. Poult. Sci., 96: 3645-3653. doi: 10.3382/ps/pex178.
Selles, SMA; Kouidri, M; Belhamiti, BT and Ait Amrane, A (2020). Chemical composition, in-vitro antibacterial and antioxidant activities of Syzygium aromaticum essential oil. J. Food Meas. Charact., 14: 2352-2358. doi: 10.1007/ s11694-020-00482-5.
Shaji, S; Selvaraj, RK and Shanmugasundaram, R (2023). Salmonella infection in poultry: a review on the pathogen and control strategies. Microorganisms. 11: 2814. doi: 10.3390/microorganisms11112814.
Sharafi, H; Moradi, M and Sharafi, K (2023). A systematic review and meta-analysis of the use of plant essential oils and extracts in the development of antimicrobial edible films for dairy application. Vet. Res. Forum. 14: 179-194. doi: 10.30466/vrf.2022.1986122.3730.
Shen, S; Zhang, T; Yuan, Y; Lin, S; Xu, J and Ye, H (2015). Effects of cinnamaldehyde on Escherichia coli and Staphylococcus aureus membrane. Food Control. 47: 196-202. doi: 10.1016/j.foodcont.2014.07.003.
Stevanović, ZD; Bošnjak-Neumüller, J; Pajić-Lijaković, I; Raj, J and Vasiljević, M (2018). Essential oils as feed additives—future perspectives. Molecules. 23: 1717. doi: 10.3390/molecules23071717.
Tugnoli, B; Giovagnoni, G; Piva, A and Grilli, E (2020). From acidifiers to intestinal health enhancers: How organic acids can improve growth efficiency of pigs. Animals. 10: 134. doi: 10.3390/ani10010134.
Turek, C and Stintzing, FC (2013). Stability of essential oils: a review. Compr. Rev. Food Sci. Food Saf., 12: 40-53. doi: 10.1111/1541-4337.12006.
Van, TTH; Smooker, PM; Wu, S and Wu, Z (2023). Bacterial diseases in poultry: Biology, virulence and prevention in the age of reduced antibiotic use. Front. Vet. Sci., 10: 1189315. doi: 10.3389/fvets.2023.1189315.
Vertiprakhov, V and Grozina, A (2019). Exocrine pancreatic function in chickens as a result of adding feed acidifiers to their diet. Sib. Herald Agric. Sci., 48: 63-69. doi: 10.26898/0370-8799-2018-6-9.
Vinolya, RE; Balakrishnan, U; Yasir, B and Chandrasekar, S (2021). Effect of dietary supplementation of acidifiers and essential oils on growth performance and intestinal health of broiler. J. Appl. Poult. Res., 30: 100179. doi: 10.1016/j.japr.2021.100179.
Visvalingam, J; Palaniappan, K and Holley, RA (2017). In vitro enhancement of antibiotic susceptibility of drug resistant Escherichia coli by cinnamaldehyde. Food Control. 79: 288-291. doi: 10.1016/j.foodcont.2017.04.011.
Waghmare, S; Gupta, M; Bahiram, K; Korde, J; Bhat, R; Datar, Y; Rajora, P; Kadam, M; Kaore, M and Kurkure, N (2025). Effects of organic acid blends on the growth performance, intestinal morphology, microbiota, and serum lipid parameters of broiler chickens. Poult. Sci., 104: 104546. doi: 10.1016/j.psj.2024.104546.
Yin, L; Dai, Y; Chen, H; He, X; Ouyang, P; Huang, X; Sun, X; Ai, Y; Lai, S and Zhu, L (2022). Cinnamaldehyde resist Salmonella typhimurium adhesion by inhibiting type I fimbriae. Molecules. 27: 7753. doi: 10.3390/molecules 27227753.

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