Fungal spoilage of starch-based foods in relation to its water activity (a w )

Journal of Stored Products Research 36 (2000) 47±54 www.elsevier.com/locate/jspr Fungal spoilage of starch-based foods in relation to its water activity (a w ) Noorlidah Abdullah a, *, A. Nawawi a, I. Othman b a Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603, Kuala Lumpur, Malaysia b Department of Biochemistry, Faculty of Medicine, University of Malaya, 50603, Kuala Lumpur, Malaysia Accepted 2 June 1999 Abstract Equilibrium moisture contents of starch-based foods were determined at nine levels of relative humidity ranging from 10% to 98%, using a static desiccator technique. From the water adsorption isotherms, the critical moisture contents (% dry basis), i.e. the moisture contents to be maintained at 258C that will not allow fungal growth were found to be 13.0% for rice grains, 12.9% for glutinous rice grains, 9.6% for rice our, 10.6% for glutinous rice our, 10.7% for wheat our and 10.5% for corn our. To maintain these moisture content levels, starch-based foods must be stored at water activity (a w ) levels of not more than 0.65. From this study, at 0.65 a w, visible appearance of fungi did not occur in ours until 6 months of storage. However, spoilage occurred after 57 days for ordinary rice grains and 73 days for glutinous rice grains. Hence, a moisture content lower than 13.0% for ordinary rice grains and 12.9% for glutinous rice grains must be maintained for long-term storage. # 2000 Elsevier Science Ltd. All rights reserved. Keywords: Adsorption isotherm; Fungal spoilage; Starch-based food; Water activity 1. Introduction Cereals, before being consumed as food, go through the processes of cultivation, harvesting, drying, preparation and marketing (including storage) all under natural conditions, and therefore, often involve microbiological contamination and infection. The water activity level in * Corresponding author. Tel.: +603-7594371; fax: +603-7594178. E-mail address: norah@botany.um.edu.my (N. Abdullah) 0022-474X/00/$ - see front matter # 2000 Elsevier Science Ltd. All rights reserved. PII: S0022-474X(99)00026-0

48 N. Abdullah et al. / Journal of Stored Products Research 36 (2000) 47±54 food is of practical importance as it controls the onset and severity of mould spoilage. It is commonly observed that foods most likely to show rapid deterioration due to biological and chemical changes are usually those with high water content. The concept of water activity (a w ) introduced by Scott (1957), is the most useful expression of the water availability for microbial growth (Lacey and Magan, 1991) and enzyme activity (Acker, 1963). Water activity is only one of the environmental factors in uencing growth of fungi. Their growth and survival will be in uenced by temperature, hydrogen ion concentration, oxygen and carbon dioxide concentrations, and the presence of preservatives (Pitt and Hocking, 1985). When any of these factors is suboptimal, the inhibitory e ect of reduced a w tends to be enhanced. The a w levels corresponding to a range of moisture contents of food may be plotted to provide a water sorption isotherm. This isotherm is useful, not only in showing at what moisture contents certain desirable or undesirable levels of a w are achieved, but also in indicating what signi cance small changes in moisture content will have in terms of a w. Hence, it is a useful guide to the storage life of foods held at moderate temperatures and preserved only by reduced a w. Growth of fungi on improperly stored food sometimes causes mycotoxins to be produced (Jimenez et al., 1991; Scudamore and Hetmanski, 1995). Water sorption isotherms of cereals and their products have been studied by many workers, for example, of rice by Rangaswamy (1973), Hunt and Pixton (1974) and Gough and King (1980) and of wheat ours, by Bushuk and Winkler (1957), Pratap et al. (1982) and Leiras and Iglesias (1991). However, no published information is available on the isotherms of glutinous rice, glutinous rice our and rice our. The present study is primarily concerned with the development of fungi on several starchbased foods stored within a limited range of water activity. The sorption isotherms of these starch-based foods are included to obtain the maximum moisture content of each food to be maintained in order to prolong the shelf-life of each food. 2. Materials and methods 2.1. Sampling and subsampling of starch-based foods The starch-based foods studied were rice, glutinous rice, rice our, glutinous rice our, wheat our and corn our. Fifty samples of each starch-based food were purchased at random from retail outlets. The our samples consisted of various brands available in the market. One hundred grams of each sample of rice, glutinous rice, rice our, glutinous rice our, wheat our and corn our were bulked and mixed separately in sterile polyethylene bags. Subsequently, small proportions from di erent parts of the bulk samples were taken out using a sterile spatula and mixed again in another closed sterile polyethylene bag. Suitable sized subsamples from each sample of starch-based food were taken for subsequent experiments. 2.2. Determination of the initial moisture content of starch-based foods The moisture contents (m.c.) of the subsamples of each starch-based food were determined

N. Abdullah et al. / Journal of Stored Products Research 36 (2000) 47±54 49 by drying a weighed ground sample of food in a mechanically ventilated oven at 808C for 3 days. The samples were weighed until constant weight was achieved. The percentage moisture content was derived on a dry weight basis. An average of six replicates were determined. 2.3. Determination of the water adsorption isotherm of starch-based food The water adsorption isotherms were determined gravimetrically by exposing food samples to atmospheres of known equilibrium relative humidities ranging from 10% to 98%. Ten grams of sub-samples of starch-based food were placed in glass Petri dishes and conditioned to a speci c level of water activity in a desiccator containing an aqueous solution of suitable concentrations of sodium hydroxide (analytical grade) or a solid salt of potassium dichromate (analytical grade) in contact with its saturated solution according to Stokes and Robinson (1949). The volume of the solution used was large enough so that moisture lost or gained by the food being conditioned did not alter the composition of the controlling solution. The desiccators were kept in constant temperature incubators at 2520.58C. The speci c gravities of the solutions were measured at the beginning and at the end of each conditioning experiment and no change was observed. To ensure moisture absorption in each case, the samples were rst dried to <3% moisture content. This drying was done at room temperature using activated silica gel. It took about 2± 4 days for this drying. They were then irradiated with ultraviolet light for 30 min according to Leiras and Iglesias (1991) in order to prevent or delay microbial spoilage of samples at high relative humidity. Preliminary experiments showed that visible fungal growth occurred after equilibration of the samples for 5±14 days. Equilibrium moisture contents of the samples were then determined as above, on a dry weight basis. The experiment was repeated to obtain triplicate data. 2.4. Fungal development in starch-based foods stored at various levels of water activity The time taken for fungal development at various levels of a w was determined by storing 25 g subsamples in desiccators having relative humidities ranging from 65% to 98% as described above. The samples were examined periodically under a binocular microscope (40 magni cation) for the appearance of fungi. The number of days before visible fungal growth was noted for each starch-based food stored for a period of 6 months at the various water activities. The experiment was repeated three times. 3. Results and discussions 3.1. The initial moisture content of starch-based foods The initial moisture content (% dry basis) of each subsample of starch-based foods investigated showed that glutinous rice grains had a higher average moisture content (10.1%) than ordinary rice grains (9.3%). Among the ours, rice our has the highest average moisture

50 N. Abdullah et al. / Journal of Stored Products Research 36 (2000) 47±54 content (6.8%) followed by wheat our (6.7%) and corn our (6.4%). Glutinous rice our has the lowest average moisture content of 5.8%. 3.2. Water adsorption isotherm of starch-based foods The equilibrium moisture content values for six starch-based food at 258C are summarized in Table 1 and shown in Figs. 1 and 2. A comparison of the equilibrium moisture content data reveals that glutinous rice grains have a higher sorptive capacity than ordinary rice grains while among the ours, wheat our has the highest sorptive capacity followed by glutinous rice our, rice our and corn our in that order. It is generally considered that fungal growth on cereals is not signi cant at a w less than 0.62±0.70 (Pitt, 1975; Gough and Bateman, 1977; Gough and King, 1980; Hill and Lacey, 1983). Hence, the moisture content corresponding to this a w range is of particular interest as the critical moisture content for safe storage. Thus, from Table 1, taking a w of 0.65 as the safe storage level, the critical moisture content (% dry basis) of ordinary rice grains at 258C is 13.0%, of glutinous rice grains is 12.9%, of rice our is 9.6%, of glutinous rice our is 10.6%, of wheat our is 10.7% and of corn our is 10.5%. The adsorption isotherm curves given by Figs. 1 and 2, characterize the equilibrium state between the moisture content of starch-based food and the water vapour pressure of the surrounding atmosphere, the equilibrium being reached by gaseous exchange. On each graph, region A corresponds to the adsorption of a monomolecular lm of water, region B to adsorption of additional layers over this monolayer; and region C to condensation of water in the pores of the material followed by dissolution of the soluble material present for each starch-based food. In accordance with other studies on isotherms of cereals (e.g. Christian, 1980; Leiras and Iglesias, 1991), the starch-based foods investigated displayed a type II isotherm, having a typical sigmoidal shape. The initial average moisture contents (% dry basis) of the composite samples of each starch- Table 1 Equilibrium moisture contents of starch-based foods over a range of water activities at 258C. Average of three determinations Equilibrium moisture content (% dry basis) a w Rice Glutinous rice Rice our Glutinous rice our Wheat our Corn our 0.98 21.93 25.57 20.87 23.63 23.11 20.69 0.95 20.73 24.02 19.48 21.19 22.66 19.43 0.85 16.43 16.92 15.28 15.42 16.11 15.60 0.75 14.13 14.21 11.18 11.83 12.44 12.70 0.65 13.01 12.87 9.56 10.61 10.72 10.52 0.50 11.79 11.38 8.18 8.53 8.59 8.80 0.35 10.34 9.93 7.09 6.42 7.11 6.75 0.20 8.62 8.00 5.79 4.47 5.32 5.43 0.10 7.11 6.36 4.22 3.68 4.08 3.85

N. Abdullah et al. / Journal of Stored Products Research 36 (2000) 47±54 51 Fig. 1. Water adsorption isotherms for ordinary rice, glutinous rice, rice our and glutinous rice our at 258C. based food when sampled randomly at various retail outlets were found to be below their respective critical moisture contents. As shown in Figs. 1 and 2, all these moisture contents correspond to water activities in either region A or B whereby water is loosely or tightly bound Table 2 Days before visible appearance of fungi on six starch-based food at 258C. Average of three determinations a w Rice Glutinous rice Rice our Glutinous rice our Wheat our Corn our 0.98 722 722 1123 1422 821 2223 0.95 921 922 1524 1724 1021 5122 0.90 1022 1023 2021 2423 1021 6922 0.85 1022 1020 2721 3922 1423 12424 0.80 1321 1721 2823 6424 1721 a 0.75 2022 1921 3221 9123 2722 a 0.65 5722 7321 a a a a a No fungal development at 6 months incubation.

52 N. Abdullah et al. / Journal of Stored Products Research 36 (2000) 47±54 Fig. 2. Water adsorption isotherms for wheat our and corn our at 258C. and is unavailable for reaction as de ned by Labuza (1968), Duckworth (1975) and Troller and Christian (1978). The critical moisture content can vary from one foodstu to another and is estimated to be 14.5% for sorghum and 12.5±13.5% for wheat and maize (Christensen, 1973), but as low as 8% for groundnuts (McDonald, 1968). The critical moisture content varies with the kind of grain (Hunt and Pixton, 1974; Christian, 1980) and even variety (Gough and King, 1980). 3.3. Time taken for the development of fungi in starch-based foods stored at various levels of water activity at 258C The average numbers of days before visible fungal development on six starch-based foods are given in Table 2. At 0.75 a w, the data indicated a shelf-life of about 20 days for ordinary rice and glutinous rice grains, about 1 month for rice our and wheat our, 3 months for glutinous rice our whilst corn our can be stored for much longer than 6 months.

N. Abdullah et al. / Journal of Stored Products Research 36 (2000) 47±54 53 At 0.65 a w, fungal growth on rice and glutinous rice, while not entirely prevented, took place after a period of 2 months or more. No fungal growth occurred on rice our, glutinous rice our and wheat our when stored for 6 months. Corn our can be safely stored at 0.80 a w for 6 months. The susceptibility of foodstu s to fungal growth depends on several factors, of which the water activity and the length of storage are most important. In this study, at 258C, for short term storage (i.e. <90 days), it is recommended that ordinary rice, glutinous rice, rice our and wheat our be stored at a w of <0.65 while glutinous rice our can be maintained at a higher a w of 0.75 and corn our at 0.85 a w (Table 1). A somewhat larger safety margin is advisable for long storage periods. Barton-Wright and Tomkins (1940) indicated that fungal growth in our was prevented for 4 months at 0.79 a w and in bran at 0.75 a w. Snow et al. (1944) recommended a water activity not exceeding 0.72 for 3 months storage and not above 0.65 when storage for 2±3 years was required. In tropical conditions, a water activity of 0.7 is not low enough (Scott, 1957). Di erences were observed in the time taken for the appearance of fungi on the di erent starch-based foods stored at the same water activity. From this study, fungal growth occurred on rice and glutinous rice grains at a w of 0.65 whereas their corresponding ours did not develop fungal growth at the same a w even after a period of 6 months. Flours being highly processed foodstu s, have a lower sorptive capacity than grains at low water activities (Table 1) and greatly reduced viable spore numbers. Hence, the concentration of the initial inoculum and the physiological properties of the di erent varieties of fungal species occurring in the di erent samples might have a ected the rate of development of moulding. Also, the type of starch-based food, particularly the availability of the soluble nutrients, might have a ected the time of germination of fungal spores and the production of fungal mycelium. It has been shown by Snow et al. (1944) that fungi developed most rapidly on samples where a balance of nutrients was provided, i.e. the starch and protein mixtures and also food-borne fungi grow more vigorously in media rich in carbohydrates (Pitt and Hocking, 1977). The a w of ordinary rice at 13.0% moisture content and glutinous rice at 12.9% moisture content is 0.65 whereas at a w of 0.75 the moisture content of ordinary rice and glutinous rice is 14.1% and 14.2% respectively. Ordinary rice and glutinous rice at 0.75 a w may spoil in about 20 days but at 0.65 a w, it would be safe for about 2 months. Hence, an error in the moisture content of 1.1% for rice and 1.3% for glutinous rice is disastrous. Similarly, rice our, glutinous rice our and wheat our at a w of 0.65 and corn our at a w of 0.80 is safe for 6 months but at a w of 0.75 rice our and wheat our will spoil at about 1 and 3 months for glutinous rice our. Corn our on the other hand, will spoil at 4 months at 0.85 a w. Hence, knowledge of water adsorption isotherms of foods will indicate the shelf-life of foods and the desirable moisture content of each food at which it should be maintained. Acknowledgements The authors are grateful to the Public Service Department, Malaysia for sponsoring this project and University of Malaya for granting Noorlidah Abdullah study leave under the Academic Sta Training Scheme.

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