Modeling the Effects of Citrus sinensis Essential Oil and Nitrite on Growth Probability of Clostridium botulinum Type A in Broth Media

Author(s): Mohammad A. Rezaei, Vadood Razavilar*, Amirali Anvar, Zohreh Mashak

Journal Name: Current Nutrition & Food Science

Volume 16 , Issue 7 , 2020

Become EABM
Become Reviewer
Call for Editor

Graphical Abstract:


Background: Nitrite is a multifunctional food additive used for control of toxigenic Clostridium botulinum in foodstuffs. However, there is a growing concern about the carcinogenic and teratogenic effects of nitrite. The present research was done to assess the effects of Citrus sinensis essential oil and nitrite on the growth probability of C. botulinum type A using predictive mathematical modeling technique in broth media.

Methods: Essential oil of C. sinensis was collected using Clevenger. Multifactorial design included diverse C. sinensis, nitrite and NaCl concentrations and also different pH ranges and storage temperatures were arranged in BHI broth medium. C. botulinum type A strains were then inoculated and their growth model was analyzed.

Results: The synergistic inhibitory effects of nitrite and C. sinensis were significant (P<0.05). C. sinensis (0.045%) and nitrite (20 ppm) strongly decreased the growth of C. botulinum (log P%= - 2.2 versus log P%= 1.15). Decreasing temperature up to 25°C significantly affected growth probability of C. botulinum (P<0.05). Increasing NaCl concentration up to 3% did not cause any significant differences in the growth of C. botulinum (P= 0.062). Bacterial growth in broth media was completely inhibited at pH 5.5 and also in media contained C. sinensis (0.045%) and nitrite (60 ppm) at pH 6.5 (log P%= -3.76).

Conclusion: Using certain concentrations of C. sinensis essential oil with other suboptimal factors (pH and temperature) and nitrite can control the growth of C. botulinum in broth media.

Keywords: Citrus sinensis, Clostridium botulinum, growth probability, nitrite, broth media, modeling.

Johnson EA. Control of Clostridium botulinum in foods. In: New Weapons to Control Bacterial Growth. Villa TG, Vinas M, Eds. Switzerland: Springer 2016; pp. 83-93.
Kull S, Schulz KM, Weisemann J, et al. Isolation and functional characterization of the novel Clostridium botulinum neurotoxin A8 subtype. PLoS One 2015; 10(2): e0116381.
[] [PMID: 25658638]
Zhang JC, Sun L, Nie QH. Botulism, where are we now? Clin Toxicol (Phila) 2010; 48(9): 867-79.
[] [PMID: 21171845]
Lee S, Lee H, Kim S, et al. Microbiological safety of processed meat products formulated with low nitrite concentration - A review. Asian-Australas J Anim Sci 2018; 31(8): 1073-7.
[] [PMID: 29531192]
Glass KA, Johnson EA. Formulating Low-Acid Foods for Botulinal Safety. In: Control of Foodborne Organisms. Juneja VK, Sofos JN, Eds. New York: Marcel Dekker 2001; pp. 323-50.
Bedale W, Sindelar JJ, Milkowski AL. Dietary nitrate and nitrite: benefits, risks, and evolving perceptions. Meat Sci 2016; 120: 85-92.
[] [PMID: 26994928]
Ma L, Hu L, Feng X, Wang S. Nitrate and nitrite in health and disease. Aging Dis 2018; 9(5): 938-45.
[] [PMID: 30271668]
Dutra MP. Use of gamma radiation on control of Clostridium botulinum in mortadella formulated with different nitrite levels. Radiat Phys Chem 2016; 119: 125-9.
Jin SK, Choi JS, Yang HS, Park TS, Yim DG. Natural curing agents as nitrite alternatives and their effects on the physicochemical, microbiological properties and sensory evaluation of sausages during storage. Meat Sci 2018; 146: 34-40.
[] [PMID: 30086439]
Alahakoon AU, Jayasena DD, Ramachandra S, Jo C. Alternatives to nitrite in processed meat: up to date. Trends Food Sci Technol 2015; 45(1): 37-49.
Hąc-Wydro K, Flasiński M, Romańczuk K. Essential oils as food eco-preservatives: Model system studies on the effect of temperature on limonene antibacterial activity. Food Chem 2017; 235: 127-35.
[] [PMID: 28554616]
Wang J, Chen D, Lei Y, et al. Citrus sinensis annotation project (CAP): a comprehensive database for sweet orange genome. PLoS One 2014; 9(1): e87723.
[] [PMID: 24489955]
Fisher K, Phillips CA. The effect of lemon, orange and bergamot essential oils and their components on the survival of Campylobacter jejuni, Escherichia coli O157, Listeria monocytogenes, Bacillus cereus and Staphylococcus aureus in vitro and in food systems. J Appl Microbiol 2006; 101(6): 1232-40.
[] [PMID: 17105553]
Toscano-Garibay JD, Arriaga-Alba M, Sánchez-Navarrete J, et al. Antimutagenic and antioxidant activity of the essential oils of Citrus sinensis and Citrus latifolia. Sci Rep 2017; 7(1): 11479.
[] [PMID: 28904369]
Garre A, Fernández PS, Lindqvist R, Egea JA. Bioinactivation: software for modelling dynamic microbial inactivation. Food Res Int 2017; 93: 66-74.
[] [PMID: 28290281]
Basti AA, Misaghi A, Khaschabi D. Growth response and modelling of the effects of Zataria multiflora Boiss. essential oil, pH and temperature on Salmonella typhimurium and Staphylococcus aureus. LWT-Food Sci Techno 2007; 40(6): 973-81.
Ferhat MA, Meklati BY, Chemat F. Comparison of different isolation methods of essential oil from Citrus fruits: cold pressing, hydrodistillation and microwave ‘dry’ distillation. Flavour Fragrance J 2007; 22(6): 494-504.
Whiting RC, Strobaugh TP. Expansion of the time-to-turbidity model for proteolytic Clostridium botulinum to include spore numbers. Food Microbiol 1998; 15(4): 449-53.
Khanjari A, Misaghi A, Basti AA, et al. Effects of Zataria multiflora Boiss. Essential Oil, nisin, pH and temperature on Vibrio parahaemolyticus ATCC 43996 and its thermostable direct hemolysin production. J Food Saf 2013; 33(3): 340-7.
Razavilar V, Genigeorgis C. Prediction of Listeria spp. growth as affected by various levels of chemicals, pH, temperature and storage time in a model broth. Int J Food Microbiol 1998; 40(3): 149-57.
[] [PMID: 9620122]
Azar AP, Nekoei M, Larijani K, Bahraminasab S. Chemical composition of the essential oils of Citrus sinensis cv. valencia and a quantitative structure-retention relationship study for the prediction of retention indices by multiple linear regression. J Serb Chem Soc 2011; 76(12): 1627-37.
Ferhat MA, Meklati BY, Smadja J, Chemat F. An improved microwave Clevenger apparatus for distillation of essential oils from orange peel. J Chromatogr A 2006; 1112(1-2): 121-6.
[] [PMID: 16384566]
Hosni K, Zahed N, Chrif R, et al. Composition of peel essential oils from four selected Tunisian citrus species: evidence for the genotypic influence. Food Chem 2010; 123(4): 1098-104.
Minh Tu NT, Thanh LX, Une A, Ukeda H, Sawamura M. Volatile constituents of Vietnamese pummelo, orange, tangerine and lime peel oils. Flavour Fragrance J 2002; 17(3): 169-74.
Raut JS, Karuppayil SM. A status review on the medicinal properties of essential oils. Ind Crops Prod 2014; 62: 250-64.
Settanni L, Palazzolo E, Guarrasi V, et al. Inhibition of foodborne pathogen bacteria by essential oils extracted from citrus fruits cultivated in Sicily. Food Control 2012; 26(2): 326-30.
Randazzo W, Jiménez-Belenguer A, Settanni L, et al. Antilisterial effect of citrus essential oils and their performance in edible film formulations. Food Control 2016; 59: 750-8.
Ismaiel AA, Pierson MD. Effect of sodium nitrite and origanum oil on growth and toxin production of Clostridium botulinum in TYG broth and ground pork. J Food Prot 1990; 53(11): 958-60.
[] [PMID: 31022781]
Nevas M, Korhonen A-R, Lindström M, Turkki P, Korkeala H. Antibacterial efficiency of Finnish spice essential oils against pathogenic and spoilage bacteria. J Food Prot 2004; 67(1): 199-202.
[] [PMID: 14717375]
Khanzadi S, Gharibzadeh S, Raoufy MR, Razavilar V, Khaksar R, Radmehr B. Application of artificial neural networks to predict Clostridium botulinum growth as a function of Zataria multiflora essential oil, pH, NaCl and temperature. J Food Saf 2010; 30(2): 490-505.
Chaibi A, Ababouch LH, Belasri K, Boucetta S, Busta FF. Inhibition of germination and vegetative growth of Bacillus cereus T and Clostridium botulinum 62A spores by essential oils. Food Microbiol 1997; 14(2): 161-74.
Chouhan S, Sharma K, Guleria S. Antimicrobial activity of some essential oils-present status and future perspectives. Medicines (Basel) 2017; 4(3): 58.
[] [PMID: 28930272]
Jensen MJ, Genigeorgis C, Lindroth S. Probability of growth of Clostridium botulinum as affected by strain, cell and serologic type, inoculum size and temperature and time of incubation in a model broth system. J Food Saf 1986; 8(2): 109-26.
Baker DA, Genigeorgis C, Glover J, Razavilar V. Growth and toxigenesis of C. botulinum type E in fishes packaged under modified atmospheres. Int J Food Microbiol 1990; 10(3-4): 269-89.
[] [PMID: 2204405]
Lund BM, Peck MW. Clostridium botulinum Guide to foodborne pathogens. 2nd ed. Oxford: John Wiley & Sons 2013; pp. 91-111.
Zhao L, Montville TJ, Schaffner DW. Inoculum size of Clostridium botulinum 56A spores influences time‐to‐detection and percent growth‐positive samples. J Food Sci 2000; 65(8): 1369-75.
Lalitha KV, Gopakumar K. Combined effect of sodium chloride, pH and storage temperature on growth and toxin production by Clostridium botulinum. J Aquat Food Prod 2007; 16(2): 27-39.
Khanipour E, Flint SH, McCarthy OJ, et al. Modelling the combined effects of salt, sorbic acid and nisin on the probability of growth of Clostridium sporogenes in a controlled environment (nutrient broth). Food Control 2016; 62: 32-43.
Ripolles-Avila C, Hascoët S, Guerrero-Navarro E, Rodríguez-Jerez J. Establishment of incubation conditions to optimize the in vitro formation of mature Listeria monocytogenes biofilms on food-contact surfaces. Food Control 2018; 92: 240-8.
Cui H, Gabriel AA, Nakano H. Antimicrobial efficacies of plant extracts and sodium nitrite against Clostridium botulinum. Food Control 2010; 21(7): 1030-6.
Safarpoor FD, Gandomi H, Basti AA, Misaghi A, Rahimi E. Phenotypic and genotypic characterization of antibiotic resistance of methicillin-resistant Staphylococcus aureus isolated from hospital food. Antimicrob Res Infect Control 2017; 6: 104.
Safarpoor Dehkordi F, Akhondzadeh Basti A, Gandomi H, Misaghi A, Rahimi E. Pathogenic Staphylococcus aureus in hospital food samples; prevalence and antimicrobial resistance properties. J Food Safety 2018; 38(6): e12501.
Ranjbar R, Safarpoor Dehkordi F, Sakhaei Shahreza MH, Rahimi E. Prevalence, identification of virulence factors, O-serogroups and antibiotic resistance properties of Shiga-toxin producing Escherichia coli strains isolated from raw milk and traditional dairy products. Antimicrob Resist Infect Control 2018; 7: 53.
Ranjbar R, Masoudimanesh M, Dehkordi FS, Jonaidi-Jafari N, Rahimi E. Shiga (Vero)-toxin producing Escherichia coli isolated from the hospital foods; virulence factors, o-serogroups and antimicrobial resistance properties. Antimicrob Res Infect Control 2017; 6: 4.
Hemmatinezhad B, Khamesipour F, Mohammadi M, Safarpoor Dehkordi F, Mashak Z. Microbiological investigation of O serogroups, virulence factors and antimicrobial resistance properties of shiga toxin‐producing Escherichia coli isolated from ostrich, turkey and quail meats. J Food Safety 2015; 35: 491-500.
Momtaz H, Safarpoor Dehkordi F, Taktaz T, Rezvani A, Yarali S. Shiga toxin-producing Escherichia coli isolated from bovine mastitic milk: serogroups, virulence factors, and antibiotic resistance properties. Sci World J 2012; 2012: 618709.
Dehkordi FS, Parsaei P, Saberian S, et al. Prevalence study of Theileria annulata by comparison of four diagnostic techniques in southwest Iran. Bulgar J Vet Med 2012; 15: 123-30.
Momtaz H, Rahimian MD, Safarpoor Dehkordi F. Identification and characterization of Yersinia enterocolitica isolated from raw chicken meat based on molecular and biological techniques. J Appl Poult Res 2013; 22: 137-45.
Safarpoor Dehkordi F, Barati S, Momtaz H, Hosseini Ahari SN, Nejat Dehkordi S. Comparison of shedding, and antibiotic resistance properties of Listeria monocytogenes isolated from milk, feces, urine, and vaginal secretion of bovine, ovine, caprine, buffalo, and camel species in Iran. Jundishapur J Microbiol 2013; 6: 284-94.
Rahimi E, Yazdanpour S, Dehkordi FS. Detection of Toxoplasma gondii antibodies in various poultry meat samples using enzyme linked immuno sorbent assay and its confirmation by polymerase chain reaction. J Pure Appl Microbiol 2014; 8: 421-7.

Rights & PermissionsPrintExport Cite as

Article Details

Year: 2020
Page: [1064 - 1071]
Pages: 8
DOI: 10.2174/1573401315666191003100702
Price: $65

Article Metrics

PDF: 22