In this study, 20 samples from three different sources of animal feed were investigated and six bacterial isolates were identified. The susceptibility of four natural antimicrobials, namely, eugenol, cinnamaldehyde, thymol, and carvacrol, against six of these isolates was determined. Carvacrol and eugenol showed better inhibitory effects with larger zones of inhibition. The minimal inhibitory concentration for a range of antibiotics on the susceptibility of two isolates (namely, Sphingomonas paucimobilis and Klebsiella oxytoca) was investigated using the VITEK® 2 microbiological identification system. Both isolates showed a variety of resistance to 18 antibiotics. The minimal inhibitory concentration and fractional inhibitory concentration index of those two isolates for ampicillin and nitrofurantoin in combination with four phenolic compounds was determined. Synergistic interactions were found for most antimicrobial/antibiotic combinations; thymol and carvacrol were very effective (fractional inhibitory concentration ≤0.5) in combination with all antibiotics tested against S. paucimobilis and K. oxytoca, respectively. Ultra performance liquid chromatography techniques were used to investigate the physiological effects of the four natural antimicrobials against those two isolates. Two identical peaks were found to be systematically different between cinnamaldehyde-treated and -untreated cells. The identity of the peaks is unknown and further investigation is needed.
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References
1.
AliskanH, CanF, DemirbilekM, ColakogluS, KilicS, ArslanH. Determining in vitro synergistic activities of tigecycline with several other antibiotics against Brucella melitensis using checkerboard and time-kill assays. 18th European Congress of Clinical Microbiology and Infectious Diseases/26th International Congress of Chemotherapy, April 19–22, 2008, Barcelona, Spain. J Chemother, 2009; 21:24–30.
2.
Anonymous. Antimicrobial use in animal feed–time to stop. N Engl J Med, 2001; 345:1202–1203.
3.
BagamboulaCF, UyttendaeleM, DebevereJ. Inhibitory effect of thyme and basil essential oils, carvacrol, thymol, estragol, linalool and p-cymene towards Shigella sonnei and S. flexneri. Food Microbiol, 2004; 21:33–42.
4.
BrackmanG, DefoirdtT, MiyamotoC, BossierP, CalenberghSV, NelisH, CoenyeT. Cinnamaldehyde and cinnamaldehyde derivatives reduce virulence in Vibrio spp. by decreasing the DNA-binding activity of the quorum sensing response regulator LuxR. BMC Microbiol, 2008; 8:149.
5.
BrisseS, van DuijkerenE. Identification and antimicrobial susceptibility of 100 Klebsiella animal clinical isolates. Veterinary Microbiol, 2005; 105:307–312.
6.
BurtS. Essential oils: their antibacterial properties and potential applications in foods: a review. Int J Food Microbiol, 2004; 94:223–253.
7.
BurtS, VlielanderR, HaagasmanHP, VeldhuizenEJA. Increase in activity of essential oil components carvacrol and thymol against Escherichia coli O157:H7 by addition of food stabilizers. J Food Prot, 2005; 68:919–926.
8.
[CLSI] Clinical and Laboratory Standards Institute. Performance standards for antimicrobial disk and dilution susceptibility tests for bacteria isolated from animals; approved standard. M31-A2:1–80. Wayne, PA: NCCLS, 2002.
9.
DemoM, OlivaM, RamosB, ZigadloJ. Determination of the antibacterial activity of the pure components of essential oils. Hig Aliment, 2001; 85:87–90.
10.
DormanHJD, DeansSG. Antimicrobial agents from plants: antibacterial activity of plant volatile oils. J Appl Microbiol, 2000; 88:308–316.
11.
EliopoulosGM, MoelleringRC. Antimicrobial combinations. Antibiotics in Laboratory Medicine, 3rdLorianV. Baltimore: Williams & Wilkins, 1991; 432.
12.
FriedmanM, HenikaPR, MandrellRE. Bactericidal activities of plant essential oils and some of their isolated constituents against Campylobacter jejuni, Escherichia coli, Listeria monocytogenes and Salmonella enterica. J Food Prot, 2002; 65:1545–1560.
13.
GalloisM, RothkotterHJ, BaileyM, StokesCR, OswaldIP. Natural alternatives to in-feed antibiotics in pig production: can immunomodulators play a role?Animal, 2009; 12:1644–1661.
14.
GB/T4789.28-2003. Food sanitation microbiological test Gram staining methods, culture media and reagents. The Ministry of Public Health of People's: Republic of China, 2003; 193–231.
15.
HelanderIM, AlakomiHL, Latva-KalaK, Mattila-SandholmT, PolI, SmidEJ, GorrisLGM, Von-WrightA. Characterization of the action of selected essential oil components on Gram-negative bacteria. J Agric Food Chem, 1998; 46:3590–3595.
16.
HemaiswaryaS, KruthiventiAK, DobleM. Synergism between natural products and antibiotics against infectious diseases. Phytomedicine, 2008; 15:639–652.
17.
JacobME, FoxJT, ReinsteinSL, NagarajaTG. Antimicrobial susceptibility of foodborne pathogens in organic or natural production systems: an overview. Foodborne Pathog Dis, 2008; 6:721–730.
18.
JindalA, KocherginskayaS, MehboobA, RobertM, MackieRI, RaskinL, ZillesJL. Antimicrobial use and resistance in swine waste treatment systems. Appl Environ Microbiol, 2006; 72:7813–7820.
19.
KhachatouriansGG. Agricultural use of antibiotics and the evolution and transfer of antibiotic-resistant bacteria. Can Med Assoc J, 1998; 159:1129–1136.
20.
KobilinskyA, NazerAI, Dubois-BrissonnetF. Modeling the inhibition of Salmonella typhimurium growth by combination of food antimicrobials. Int J Food Microbiol, 2007; 115:95–109.
21.
LaiCC, TengLJ, HsuehPR, YuanA, TsaiKC, TangJL, TienHF. Clinical and microbiological characteristics of Rhizobium radiobacter infections. Clin Infectious Dis, 2004; 38:149–153.
22.
LambertRJW, SkandamisPN, CooteP, NychasG-JE. A study of the minimum inhibitory concentration and mode of action oregano essential oil, thymol and carvacrol. J Appl Microbiol, 2001; 91:453–462.
23.
LangfieldRD, ScaranoF, HeitzmanME, KondoM, HammondGB, NetoCC. Use of a modified microplate bioassay method to investigate antibacterial activity in the Peruvian medicinal plant Peperomia galiodes. J Ethnopharmacol, 2004; 94:279–281.
24.
LiJE, NieSP, QiuZH, CheMJ, LiC, XieMY. Antimicrobial and antioxidant activities of the essential oil from Herba Moslae. 14th World Congress of Food Science and Technology, 2009, Shanghai, P.R. China. J Sci Food Agric, 2010; 90:1347–1352.
25.
LinJN, LaiCH, ChenYH, LinHL, HuangCK, ChenWF, WangJL, ChungHC, LiangSH, LinHH. S. paucimobilis bacteremia in humans: 16 case reports and a literature review. J Microbiol Immunol Infection, 2010; 43:35–42.
26.
MarshallSA, JonesRN. Spectrum and antimicrobial activity of alexomycin (PNU-82, 127), a peptide compound projected for use in animal health. Diagn Microbiol Infection Dis, 1999; 33:181–186.
27.
MellonM, BenbrookC, BenbrookKL. Hogging it: estimates of antimicrobial abuse in livestock. Cambridge, Mass: Union of Concerned Scientists, 2001.
28.
MessaoudiA, GtariM, BoudabousA, WagenlehnerF. Identification and susceptibility of Klebsiella and Enterobacter spp. isolated from meat products. Afr J Microbiol Res, 2009; 3:362–369.
29.
NiuC, AfreS, GilbertES. Subinhibitory concentrations of cinnamaldehyde interfere with quorum sensing. Lett Appl Microbiol, 2006; 43:489–494.
OddsFC. Synergy, antagonism and what the chequerboard puts between them. J Antimicrob Chemother, 2003; 52:1.
32.
OlasupoNA, FitzgeraldDJ, GassonMJ, NarbadA. Activity of natural antimicrobial compounds against Escherichia coli and Salmonella enterica serovar Typhimurium. Lett Appl Microbiol, 2003; 36:448–451.
33.
OyarzabalOA. Multidrug resistance among foodborne pathogens: measures to monitor and control their appearance in poultry meat. 15th Congress of the Federation of Asian Veterinary Associations, FAVA-OIE Joint Symposium on Emerging Diseases Proceedings, Bangkok, Thailand, 2008; S45–S47.
34.
PeiRS, ZhouF, JiBP, XuJl. 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.
35.
PillaiSK, MoelleringRC, EliopoulosGM. Antimicrobial combinations. Antibiotics in Laboratory Medicine. LorianV. Philadelphia: Lippincot Williams & Wilkins, 2005; 851.
36.
Santiesteban-LopezA, PalouE, Lopez-MaloA. Susceptibility of food-borne bacteria to binary combinations of antimicrobials at selected a(w) and pH. J Appl Microbiol, 2007; 102:486–497.
37.
SiH, HuJ, LiuZ, ZengZ. Antibacterial effect of oregano essential oil alone and in combination with antibiotics against extended-spectrum β–lactamase-producing Escherichia coli. FEMS Immunol Med Microbiol, 2008; 53:190–194.
38.
SinghA, SinghRK, BhuniaAK, SinghN. Efficacy of plant essential oils as antimicrobial agents against Listeria monocytogenes in hotdogs. Lebensm Wiss Technol, 2003; 36:787–794.
39.
StockI, WiedemannB. Natural antibiotic susceptibility of Klebsiella pneumoniae, K.oxytoca, K.planticola, K.ornithinolytica and K.terrigena strains. J Med Microbiol, 2001; 50:396–406.
40.
UlteeA, BennikMHJ, MoezelaarR. The phenolic hydroxyl group of carvacrol is essential for action against the food-borne pathogen Bacillus cereus. Appl Environ Microbiol, 2002; 68:1561–1568.
41.
UlteeA, KetsEPW, SmidEJ. Mechanisms of action of carvacrol on the food-borne pathogen Bacillus cereus. Appl Environ Microbiol, 1999; 65:4606–4610.
42.
ValeroM, GinerMJ. Effects of antimicrobial components of essential oils on growth of Bacillus cereus INRA L2104 in and the sensory qualities of carrot broth. Int J Food Microbiol, 2005; 106:90–94.
43.
WegenerHC. Antibiotics in animal feed and their role in resistance development. Curr Opin Microbiol, 2003; 6:439–445.
44.
ZhangDL, McQuestinOJ, MellefontLA, RossT. The influence of non-lethal temperature on the rate of inactivation of vegetative bacteria in inimical environments may be independent of bacterial species. Food Microbiol, 2010; 27:453–459.
45.
ZhaoWH, HuZQ, HaraY, ShimamuraT. Inhibition of penicillinase by epigallocatechin gallate resulting in restoration of antibacterial activity of penicillin against penicillinase-producing Staphylococcus aureus. Antimicrob Agents Chemother, 2002; 46:2266–2268.
