Control of rope spore forming bacteria using carambola (Averrhoa carambola) fruit pomace powder in wheat bread preparation

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M.L. Sudha
P. Viswanath
V. Siddappa
S. Rajarathnam
M.N. Shashirekha


antimicrobial activity, Bacillus, bread, carambola fruit pomace, rope spore count


Wheat bread is a high moisture, low acid product, susceptible to spoilage by Bacillus spp., as the endospores survive the baking temperatures. Carambola (Averrhoa carambola) fruit pomace, a by-product obtained after juice extraction, has an insoluble dietary fibre content of 61.0±1.2% and is rich in antimicrobial phenolic compounds of 17.8±1.1 gallic acid equivalent/g (water extract). The anti-nutritional content of oxalic acid in the fruit is reduced to 0.15 mg/g in the pomace. Studies on the antimicrobial activity of the dried pomace powder in dimethyl-sulfoxide as determined by the disc diffusion assay showed that the Gram-positive bacteria tested were more sensitive than Gram-negative bacteria. The minimum inhibitory and minimum bactericidal concentration of Bacillus cereus and of Bacillus subtilis was 1.25 mg/ml of the dry pomace powder. Incorporation of carambola pomace powder into wheat flour for bread preparation at a 5% level resulted in a bread of acceptable sensory profile with a high desirable fibre content. The rope spore count of the bread with carambola fruit pomace powder at this level showed a count of 0.7 log10/g, an inhibition equal to that of bread with chemical preservatives added while the bread with no preservatives had a count of 8.76 log10/g at the end of 72 h when incubated at 35 °C, resulting in a product that was safe with added high fibre a desired nutrient quality.

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American Association of Cereal Chemists (AACC), 2000. Approved methods of the AACC (10th Ed.). Methods 08-01, 30-25, 44-15A, 46-10, 54-10, 54-21. AACC, St. Paul, MN, USA.
Anonymous, 2004. Water-insoluble fiber-rich fraction from the pomace of carambola. Natural Product Radiance 3: 367.
Association of Official Analytical Chemists (AOAC), 1995. AOAC official method 947.05 (16th Ed.). AOAC International, Arlington, VA, USA.
Association of Official Analytical Chemists (AOAC), 2000. AOAC official method 991.43 (17th Ed.). AOAC International, Arlington, VA, USA.
Bauer, A.W., Kirby, W.M.M., Sherris, J.C. and Turek, M., 1966. Antibiotic susceptibility testing by standardized single disc method. American Journal of Clinical Pathology 36: 493-496.
Chau, C., Chen, C. and Mao-Hsiang, L., 2004a. Characterization and physicochemical properties of some potential fibers derived from Averrhoa carambola. Nahrung. 48: 43-46.
Chau, C., Chen, C. and Lin, C., 2004b. Insoluble fiber-rich fractions derived from Averrhoa carambola: hypoglycemic effects determined by in vitro methods. LWT-Food Science and Technology 37: 331-335.
Chen, H., Rubenthaler, G.L., Leung, H.K. and Barnowki, J.D., 1988. Chemical, physical and baking properties of apple fiber compared with wheat and oat bran. Cereal Chemistry 65: 244-247.
Clarke, C.I., Schober, T.J., Dockery, P., O’Sullivan, K. and Arendt, E.K., 2004. Wheat sourdough fermentation: effects of time and acidification on fundamental rheological properties. Cereal Chemistry81: 409-417.
Dengate, S. and Ruben, A., 2002. Controlled trial of cumulative behavioural effects of a common bread preservative. Journal of Paediatrics and Child Health. 38: 373-376.
De Vuyst, L. and Vancanneyt, M., 2007. Biodiversity and identification of sourdough lactic acid bacteria. Food Microbiology24: 120-127.
Dubois, M., Gilles, K.A., Hamilton, J.K., Rebers, P.A. and Smith, F., 1956. Colorimetric method of determination of sugars and related substances. Analytical Chemistry 28: 350-356.
Ericsson, J.M. and Sherris, J.C., 1971. Antibiotic sensitivity testing: report of an international collaborative study. Acta Pathologica Microbiologica Scandinavica 217: 1-90.
Gilbert, R.J., Turnbull, P.C.B., Parry, J.M. and Kramer, J.M., 1981. B. cereus and other Bacillus species: their part in food poisoning and other clinical infections. In: Berkely, R.C.W. and Goodfellow, M. (eds.) The aerobic endospore-forming bacteria: classification and identification. Academic press Inc., London, UK, pp. 297-314.
Hassid, W.Z. and Neufeld, E.F., 1964. Quantitative determination of starch in plant tissue. In: Whistler, R.L. (ed.) Methods in carbohydrate chemistry, Vol. 4. Academic Press, New York, NY, USA, pp. 33-36.
Hansen, Å.S., 2004. Sourdough bread. In: Hui, Y.H., Meunier-Goddik, L., Hansen, Å.S., Josephsen, J., Nip, W.-K., Stanfield, P.S and Toldrá, F. (eds.) Handbook of food and beverage fermentation technology.Marcel Dekker Inc., New York, NY, USA, pp. 729-755.
Hassan, Y.I. and Bullerman, L.B., 2008. Antifungal activity of Lactobacillus paracasei ssp. tolerans isolated from a sourdough bread culture. International Journal of Food Microbiology 121: 112-115.
Hayes, W.B., 1960. Fruits growing in India. Kitabistan, Allahabad, India.
Indrani, D., Soumya, C., Jyotsna, R. and Venkateswara Rao, G., 2010. Multigrain bread – its dough rheology, microstructure, quality and nutritional characteristics. Journal of Texture Studies 41: 302-319.
Irvine, G.N. and McMullan, M.E., 1960. The ‘Remix’ baking test. Cereal Chemistry. 37: 603-613.
Katina, K., Arendt, E., Liukkonen, K.H., Autio, K., Flander, L. and Poutanen, K., 2005. Potential of sourdough for healthier cereal products. Trends in Food Science and Technology16: 104-112.
Mallock, J.G. and Cook, W.K., 1930. A volume measuring apparatus for small loaves. Cereal Chemistry 7: 307-310.
Moresco, O., Henrique, H., Queiroz, G.S., Pizzolatti, M.G. and Brighente, I.M.C., 2012 Chemical constituents and evaluation of the toxic and antioxidant activities of Averrhoa carambola leaves. Brazilian Journal of Pharmacognosy 22: 319-324.
Kafkas, E., Kosar, M., Turemis, N. and Baser, K.H.C., 2006. Analysis of sugars, organic acids and vitamin C contents of blackberry genotypes from Turkey. Food Chemistry 97: 732-736.
Kirschner, L.M. and Von Holy, A., 1989. Rope spoilage of bread. South African Journal of Science 85: 425-427.
Kramer, J.M. and Gilbert, R.J., 1989. Bacillus cereus and other Bacillus species. In: Doyle, M.P. (ed.) Foodborne bacterial pathogens. Marcel Dekker, New York, NY, USA, p. 22.
Lee, J., Kim, Y. and Shin, D., 2002. Antimicrobial synergistic effect of linolenic acid and monoglyceride against Bacillus cereus and Staphylococcus aureus. Journal of Agricultural Food Chemistry 50: 2193-2199.
Leuschner, R.G.K., O’Callaghan, M.J.A. and Arendt, E. K., 1998. Bacilli spoilage in part-baked and rebaked brown soda bread. Journal of Food Science 63: 915-918.
Lund, B.M., 1990. Foodborne disease due to Bacillus and Clostridium species. The Lancet 336: 982.
Masoodi, F.A. and Chauhan, G.S., 1998. Use of apple pomace as a source of dietary fiber in wheat bread. Journal of Food Processing and Preservation 22: 255-263.
Mentes, Ö., Ercan, R. and Akçelik, M., 2007. Inhibitor activities of two Lactobacillus strains, isolated from sourdough, against rope-forming Bacillus strains. Food Control18: 359-363.
Miller, G.I., 1959. Use of dinitro salicylic acid reagent for determination of reducing sugar. Analytical Chemistry 31: 426-428.
Nicholson, W.L., Munakata, N., Horneck, G., Melosh, H.J. and Setlow, P., 2000. Resistance of Bacillus endospores to extreme terrestrial and extraterrestrial environments. Microbiology and Molecular Biology Reviews 64: 548-572.
Palop, A., Raso, J., Pagan, R., Condon, S. and Sala, F.J., 1996. Influence of pH on heat resistance of Bacillus licheniformis in buffer and homogenised foods. International Journal of Food Microbiology 29: 1-10.
Pattison, T.L., Lindsay, D. and Von Holy, A., 2003. In vitro growth response of bread-spoilage Bacillus strains to selected natural antimicrobials. Journal of Basic Microbiology 43: 341-347.
Pepe, O., Blaiotta, G., Moschetti G., Greco, T. and Villani, F., 2003. Rope-producing strains of Bacillus spp. from wheat bread and strategy for their control by lactic acid bacteria. Applied and Environmental Microbiology 69: 2321-2329.
Ranganna, S., 1986. Hand book of analysis and quality control for fruit and vegetable products (2nd Ed.). Tata McGraw-Hill Publishing Company Limited, New Delhi, India.
Röcken, W. and Voysey, P.A., 1995. Sourdough fermentation in breadmaking. Journal of Applied Bacteriology – Symposium Supplement 79: 38S-48S .
Rodríguez-Lozano, A., Campagnoli, M., Jewel, K., Monadjemi, F. and Gaze Rosenkvist, J.E., 2010. Bacillus spp. thermal resistance and validation in soups. Current Research, Technology and Education Topics in Applied Microbiology and Microbial Biotechnology, 1: 537-544.
Rosenkvist, H. and Hansen, A., 1995. Contamination profiles and characterization of Bacillus species in wheat bread and raw materials for bread production. International Journal of Food Microbiology 26:353.
Rosenquist, H. and Hansen, A., 1998. The antimicrobial effect of organic acids, sour dough and nisin against Bacillus subtilis and Bacillus licheniformis isolated from wheat bread. Journal of Applied Microbiology 85: 621-631.
Ryan, L.A.M., Dal Bello, F. and Arendt, E.K., 2008. The use of sourdough fermented by antifungal LAB to reduce the amount of calcium propionate in bread. International Journal of Food Microbiology125: 274-278.
Sadeghi, A., 2008. The secrets of sourdough: a review of miraculous potentials of sourdough in bread shelf life. Biotechnology 7: 413-417.
Seiler, D.A.L., 1994. Preservation of bakery products. Institute of Food Science and Technology Proceedings 17: 35-40.
?eker, I.T., Ertop, M.H. and Hayta, M. (in press). Physicochemical and bioactive properties of cakes incorporated with gilaburu fruit (Viburnum opulus) pomace. Quality Assurance and Safety of Crops & Foods, DOI:
Sockett, P.N., 1991. Food poisoning outbreaks associated with manufactured foods in England and Wales 1980-1989. Communicable Disease Representative 1: R105-R109.
Smith, J.P., 1993. Bakery products. In: Parry, R.T. (ed.) Principles and applications of modified atmosphere packaging of foods. Blackie Academic and Professional, Glasgow, UK, pp. 134-169.
Spicher, G., 1983. Baked goods. In: Rehm, H.J. and Reed, G. (eds.) Biotechnology. Verlag Chemie, Weinheim, Germany, pp. 1-80.
Steel, R.G.D. and Torrie, J.H. 1980. Principles and procedures of statistics (2nd Ed.). McGraw-Hill, New York, NY, USA.
Stevenson, K.E. and Segner, W.P., 2001. Mesophilic aerobic sporeformers. In: Downes, F.P. and Ito, K. (eds.) Compendium of methods for the microbiological examination of foods (4th Ed.) American Public Health Association, Washington, DC, USA, pp. 223-227.
Sudha, M.L., Baskaran, V. and Leelavathi, K., 2007. Apple pomace as a source of dietary fiber and polyphenols and its applications on the rheological characteristics and cake making. Food Chemistry 104: 686-692.
Todd, E.C.D., 1982. Foodborne and waterborne disease in Canada – 1977. Annual summary. Journal of Food Protection 45: 865-883.
Valerio, F. Favilla, M., De Bellis, P., Sisto, A., de Candia, S. and Lavermicocca, P., 2009. Antifungal activity of strains of lactic acid bacteria isolated from a semolina ecosystem against Penicillium roqueforti, Aspergillus niger and Endomyces fibuliger contaminating bakery products. Systematic and Applied Microbiology32: 438-448.
Viljoen, C.R. and Von Holy, A., 1997. Microbial populations associated with commercial bread production. Journal of Basic Microbiology 37: 439-444.
Von Holy, A. and Allan, C., 1990. Current perspectives on rope in bread. In: Smith, M.F., Kort, M.J., Clarke, I.R. and Bush, P.B. (eds.) Proceedings of the Second National Bakery Symposium, Technikon Natal Printers, Durban, South Africa, pp. 119-125.
Voysey, P.A., 1989. Studies on rope in bread. FMBRA Bulletin 4: 41-48.
Wei, Q., Wolf-Hall, C. and Hall, C.A., 2009. Application of raisin extracts as preservatives in liquid bread and bread systems. Journal Food Science 74: 177-184.