Influence of pH and Storage Period on the Antibacterial Susceptibility of Enterotoxigenic Bacillus cereus in Pasteurized Cow Milk during Low Temperature Storage
1 Department of Food Technology, P.M.B. 2021, Kaduna Polytechnic, Nigeria
2 Department of Microbiology, Faculty of Life Sciences, Ahmadu Bello University, Zaria, West Africa Nigeria
* Corresponding author: sakeenabello2012@gmail.com
2 Department of Microbiology, Faculty of Life Sciences, Ahmadu Bello University, Zaria, West Africa Nigeria
* Corresponding author: sakeenabello2012@gmail.com
Abstract
Food preservation processes are usually applied with the aim of slowing down or preventing spoilage and pathogenic bacteria in food materials. This involves the application of one or more environmental stresses (or hurdles) in the form of extremes of temperature, pH, and the manipulation of other optimal growth conditions.Environmental stress that may result from the use of hurdles, has however been shown to play a role in the emergence of antimicrobial resistance. This study was conducted with the aim of studying the combined effects of pH, low storage temperature and storage period on the antibacterial susceptibility of enterotoxigenic Bacillus cereus isolates from pasteurized cow-milk to selected antibacterial agents. Mcfarland standard 1 (about 8.5 log units) of a confirmed B. cereus isolate from raw milk was inoculated into sterile pasteurized cow-milk samples at varying pH levels (6.3.6.4,6.7,6.8), and stored at refrigeration temperature (4-100C) for a period of 72 h. Isolates were assayed for the B. cereus diarrhoel enterotoxin haemolysin BL (HBL) production using B. cereus enterotoxin reversed passive agglutination (BCET-RPLA) toxin detection kits (OXOID) and then subjected to antibacterial susceptibility tests using the Kirby-Bauer method against 10 antimicrobial agents (OXOID). Antibacterial agents included Ampicillin (10μg), Cephalothin (30μg), Amoxicillin-clavulanic acid(20/10μg), Cefpodoxime (10μg) Ceftriaxone (30μg), Erythromycin (30μg), Amikacin (30g), Tetracycline (30μg)Ciprofloxacin (5μg) and Trimetoprim Sulfamethoxazole (1.25/23.75μg). Results revealed resistance of all isolates to Ampicillin, Amoxicillin-clavulanic acid, Cefpodoxime, Ceftiaxone and Trimetoprim Sulfamethoxazole. All isolates were sensitive to Erythromycin, and Ciprofloxacin.Varying patterns were observed for Cephalothin, where only isolates from milk at pH 6.8 stored for 72 h and which exhibited suppressed toxigenic ability, were sensitive. Similarly, all isolates showed susceptibility to Tetracycline except for isolates at pH 6.3 which showed less sensitivity (intermediate response) to the antibacterial agent over a 48 h period. Findings suggest pH and storage conditions of foods could induce varying degrees of susceptibility to antibacterial agents in some enterotoxigenic B. cereus isolates.
Keywords
B. cereus
Haemolysin BL
low temperature storage
pH
antibacterial susceptibility
References
- Agwa, O.K., Uzoigwe, C.I., and Wokoma, E.C. (2012). Incidence and antibiotic sensitivity of Bacillus cereus strains isolated from food sources in Turkey. African Journal of Biotechnology, 9(11):1641-1647.
- Andrews, J.M. and Wise, R. (2002). Susceptibility testing of Bacillus species. Journal of antimicrobial Chemotherapy, 49:1039-1046.
- Banerjee, M., Nair, G.B and Ramamurthy, T. (2011). Phenotypic & Genetic characterization of Bacillus cereus isolated from acute diarrheal patients Indian Journal of Medical Research, 133(1):88-95.
- Boor, K.J. (2006). Bacterial stress response: What doesn't kill them makes them stronger. PLOS Biology 4:23.
- Burgess, G. and Horwood, P. (2006). Development of improved molecular detection methods for Bacillus cereus toxins. A report for the Rural Industrial Research and Development Corporation, Australia.
- CLSI (Clinical and Laboratory Standards Institute) (2011). Performance Standards for Antimicrobial Susceptibility Testing; Twenty-First Informational Supplement, CLSI document M100-S21 (ISBN 1-56238-742-1). Clinical and Laboratory Standard Institute, 940 West Valley Road, Suite 1400, Wayne, Pennsylvania 19087 U.S.A. pp 70-75
- Desriac, N., Broussolle, V., Postolec, F., Mathot, A., Sohier, D., Coroller, L., and Leguerinel, I. (2013). Bacillus cereus cell response upon exposure to acid environment: toward the identification of potential biomarkers. Fronu Microbiology, 4:284. Published online October 2, 2013.
- Dikbas, N. (2010). Determination of antibiotic susceptibility and fatty acid methyl ester profiles of Bacillus cereus strains isolated from different food sources in Turkey. African Journal of Biotechnology, 9(11):1641-1647.
- Doorduyn, Y., de Boer, E., and van Pelt, W. (2008). Registratie voedselinfecties en vergiftigingenbij de Inspecievoor de Gezondheidszorg en de Voedsel en Waren Auoriteij, 2007. In: Mols,M. (2009). Bacillus cereus acid stress responses. PhD thesis, Wageningen University. P 18.
- Ebinesh, A. (2017). Bacterial stress response and cross resistance to antibiotics in the light of natural selection. Inernational of infectious Diseases and Immune Therapies:1-2
- Ebinesh, A. andKailash, T.V. (2016). Looking into antibiotics failure: a deemed threat, indeed not! Inernational Journal of Applied Research Sudis5: 1247 Fenselfeld, G. (2014). A brief history of epigenetics. Cold Spring Harbour Perspectives in Biology6(1): http://www.ncbi.nlm.nih.gov/pubmed. Retrieved 20/04/2018.
- Gohar. M., Okstad, O.A., Gilois, N., Sanchis, V., Kolsto, A.B. and Lereclus, D. (2002). Two-dimensional electrophoresis analysis of the extracellular proteome of Bacillus cereus reveals the importance of the Plc Pregulon. Proteomics, 26): 784-791.
- Hafiz, Y., Iqbal, A., Ahmad, M. and Ali, A. (2012). Antibiograms of B. cereus isolated from street vended foods in Srinagar area of Kashmir valley. Indian Journal of Field Veterinarians, 8(1):34-37.
- Holbrook, R. and Anderson, J.M. (1980). An improved selective and diagnostic medium for the isolation and enumeration of Bacillus cereus in foods. Canadian Journal of Microbiology, 26:753-759
- Hoseini, A.S.M, Motamedifar, M., Hadi, N. and Sedigh, E.S.H. (2016). Analysis of Virulence Genes Among Methicillin Resistant Staphylococcus aureus (MRSA) Strains. Jundishapur Journal of Microbiology, 7(6):e10741. doi:10.5812/jjm.10741
- Luna, V.A., Kling, D., Gulledge, J. Cannons, A.C., Amuse, P.T., and Cattani, J. (2007). Susceptibility of Bacillus anthracis, Bacillus mycoides, Bacillus pseudomycoides and Bacillus thuringiensis to 24 antimicrobials using sensitive automated microbroth dilution and test agar diffusion methods. Journal of Animicrobial Chemotherapy; 60(3):555-567.
- McMahon, W.S.Xu,J., Moore ,J.E., Blair, I.S. and McDowell, D.A. (2007). Environmental Stress and Antibiotic Resistance in Food-Related Pathogens. Applied and Environmental Microbiology, 73(1):211-217.
- MFLP-42 ( Laboratory Procedures For The Microbiological Analysis Of Foods) (2003). Isolation and Enumeration of Bacillus cereus in Foods. Http//www.hc-sc.gc.ca. Retrieved 27/7/2007.
- Mols, M. (2009). Bacillus cereus acid stress responses. PhD thesis, Wageningen University. P18.
- Mols, M., and Abee, T. (2011) Bacillus cereus responses to acid stress. Environmental Microbiology 13:2835-2843
- Mohammed, A. (2016). Occurrence of Bacillus cereus in raw milk and some dairy products in Egypt. Japanese Journal of veterinary research. 66:595-102
- Owusu-kwarteng, J., wuni, A., akabanda, f., tano-debrah, k. and Jespersen, L. (2017), Prevalence, virulence factor genes and antibiotic resistance of B. cereus sensulato isolated from dairy farms and traditional dairy products. Bio. Med. Central Microbiology 2017, 17:65 doi: 10.1186/s12866-017-0975-9
- Pujol, L., Kan-Kin-YU D., Le Marc, Y., Johnston, M., Rama-Heizard, F., Guillou, S and Membre, J. (2012). Establishing Equivalence for Microbial-Growth-Inhibitory Effects ("Iso-Hurdle Rules") by Analyzing Disparate Listeria monocytogenes Data with a Gamma-Type Predictive Model. Applied and Environmental Microbiology, 78(4):1069-1080.
- Rowan, N.J., Deans, J.O., Anderson, C.O., Gemmell, C.O., Hunter, I.S. and Chaithong, T. (2001). Putative virulence factor expression by clinical and food isolates of Bacillus spp. after growth in reconstituted infant milk formulae. Applied and Environmental Microbiology, 67:3873-3881.
- Russell, A.D. (2004). Bacterial adaption and resistance to antiseptics, disinfectants and preservatives is not a new phenomenon. Journal of Hospital Infection, 57(2):97-104.
- Schoeni, J. and Wong, A.C.L. (2005). Bacillus cereus food poisoning and its toxins Journal of Food Protection, 68:636-48.
- Senesi, S., and Gherardi, E. (2010). Production, Secretion and Biological Activity of Bacillus cereus Enterotoxins. Toxins, 2:1690-1703.
- Shafaati, M., Boroumand, M., Amiri, P. and Kazemian, H. (2016). Correlation Between qacE and qacEDelta Efflux Pump Genes, Antibiotic and Disinfectant Resistant Among Clinical Isolates of E. Coli. Recent Paenos on Anifecive Drug Discovery 11(2):189-95.
- Slamti, L. and Lereclus, D. (2002). A cell-cell signaling pepide activates the PlcR virulence regulon in bacteria of the Bacillus cereus group. EMBO Journal, 21: 4550-4550.
- Soto, S.M. (2009): relationship between virulence and antimicrobial resistance in bacteria. Reviews in medial microbiology, 20:84-90
- Stenfors-Arnesen, L.P., Fagerlund, A., and Granum, P.E. (2008). From soil to gut: Bacillus cereus and its food poisoning toxins. FEMS Microbiology Review, 32:579-606.
- Stickland, H.G., Anderson, D.W., Lilley, K.S., Griffin, J.L. and Welch, M. (2010). Mutation ofnfxB causes global changes in the physiology and metabolism of Pseudomonas aeruginosa. Journal of Proteome Research, 9:2957-67.
- Turnball, P.C.B., Sirranni, C., Samaan, M.N., Sutton, F.N., Reyes, A.E. andPeruski, J.L.F. (2004). MICs of selected antibiotics for Bacillus anhracis, Bacillus cereus, Bacillus thuringiensis, and Bacillus mycoides from a range of clinical and environmental sources as determined by the Etest of Clinical Microbiology, 42:3626-3634.
- Wijnands, L., Dufrenne, J., Zwietering, M.H, and Leusden, F.M. (2006). Spores from mesophilic Bacillus cereus strains germinate better and grow faster in simulated gastro-intestinal conditions than spores from psychrotrophic strains. Inernational Journal of Food Microbiology, 112(2):120-128.
- Wilcks, A., Hansen, B., Hendriksen, N.B. and Licht, T.R. (2006). Fate and effect of ingested Bacillus cereus spores and vegetative cells in the intestinal tract of human floor-associated rates. FMS Immunology and Medical Microbiology 46:70-77
- Whong, C.M.Z., Kwaga, J.K.P., Umoh, V.J. and Ameh, J.B. (2016). Densities of Bacillus cereus in some Nigerian foods and food ingredients. Nigerian Journal of Microbiology, 20(2):960-970.
- Whong, C.M.Z. and Kwaga, J.K.P. (2007). Antibiograms of Bacillus cereus isolates from some Nigerian foods Nigerian food Journal, 25(1):178-183.
- Yao, J.D.C., Dufresne, J., Ellen Jo Baron, James H. Jorgensen, Marie Louise Landry, and Michael Pfaller (eds). (2007). Antibacterial agents. In: Patrick R. Murray, Manual of clinical microbiology. (9th edition). ASM press, Washington DC, USA, p 1080.
How to Cite
S, B., C.M.Z, W., & I.O, A. (2020). Influence of pH and Storage Period on the Antibacterial Susceptibility of Enterotoxigenic Bacillus cereus in Pasteurized Cow Milk during Low Temperature Storage. Nigerian Journal of Microbiology, 34(1), 5025-5033. https://doi.org/10.67614/njm.2020.e3s6jtqo
B. S, W. C.M.Z, and A. I.O, "Influence of pH and Storage Period on the Antibacterial Susceptibility of Enterotoxigenic Bacillus cereus in Pasteurized Cow Milk during Low Temperature Storage," Nigerian Journal of Microbiology, vol. 34, no. 1, pp. 5025-5033, June 2020. doi: 10.67614/njm.2020.e3s6jtqo