Quality assessment of starter- produced weaning food subjected to different temperatures and ph

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1 African Journal of Food Science Vol. 6(5), pp , 15 Marc, 2012 Available online at ttp:// DOI: /AJFS ISSN Academic Journals Full Lengt Researc Paper Quality assessment of starter- produced weaning food subjected to different temperatures and ph Wakil, Serifa Monilola* and Oriola, Olasunkanmi Bukola Department of Microbiology, University of Ibadan, Ibadan, Nigeria. Accepted 2 December, 2011 Weaning is a gradual stoppage of feeding a baby wit te moter s milk and start feeding te baby wit semi-solid food suc as maize, sorgum or millet wic can be fermented to increase teir nutrient and carboydrate digestibility. Te weaning food was formulated by fortifying cereal (Sorgum) wit legume (Cowpea) wic ad been cleaned, steeped in sterile distilled water for 24 and germinated for 48 at 30 C. Te rootlet was remove d and dried in oven at 60 C for 24 and milled separately. Te product was formulated at te ratio of 70:30 and fermented wit te use of starter, Saccaromyces cerevisiae and Lactobacillus plantarum in single and in mixed culture Lactobacillus Saccaromyces. Te formulated blends were incubated at different temperatures and ph adjusted by te use of pospate buffer and potentiometric confirmation. Te analysis of te effect of temperature on nutrient composition sowed tat crude increases wit fermentation wile te eter extract, crude and as s decreased. Te igest crude (19.5%) was recorded at 37 C in mixed culture gruel. Te vitamin al so increase wit temperature (25 to 37 C) and decreases at 40 C and te igest vitamin w as observed at 37 C in L. Saccaromyces blend. Te vitamin increased wit fermentation and ph, and te igest was observed at ph 6.Te optimum environmental conditions for te development of nutritious weaning food is terefore a fermentation temperature of 37 C at ph 6 for 48 w it a starter combination of L. plantarum and S. cerevisiae. Key words: Weaning food, starter culture, fermentation, temperature and p canges, nutritional composition. INTRODUCTION In many West African countries, exclusive breast feeding is usually adequate up to tree to four monts of age, but after tis period it may become increasing inadequate to support te nutritional demands of te growing infants. Tus, in a weaning process tere is always a need to introduced soft, easily swallowed foods to supplement te infant s feeding early in life (Onofiok et al., 1992). Weaning foods are traditionally composed of staple cereals and legumes prepared eiter individually or as composite gruels. Cereal form basis for most of te traditional weaning foods in West Africa. Te *Corresponding autor. Semowak@yaoo.com. Tel of maize and sorgum is of poor quality, low in lysine and tryptopan. Tese two amino acids are indispensable to te growt of te young cild (Oyenuga 1968). Cereal gruel processing metods ave resulted in loss of te original tryptopan in corn during te processing of ogi (Makinde and Lacance, 1976). Indeed, Akinrele and Edward (1971) concluded tat te of ogi was too low even to support te growt of rat. Anoter report noted tat corn gruel can provide some energy, but not oter nutrients needed for te growt of te baby (Ketiku and Ayoku, 1984). Food staples suc as maize, sorgum, millet, rice can be fermented to increase te nutrient, carboydrate digestibility, and energy densities of gruels, increase te bio-availability of amino acids and also improve teir self life under controlled environment.

2 148 Afr. J. Food Sci. Te product may be consumed by gelatinizing it into stiff gel or made into gruel or pap tat is popular amongst infant as weaning food and breakfast meal amongst adults (Ozo and Kuyanbana, 1995). Many researcers ave worked extensively on cereal- legume combinations (Onilude et al., 1999; Sanni et al., 1999; Aci, 2005; Wakil and Onilude, 2009) wic ave been used to improve te quality of cereals. Spontaneous fermentation tat is, process initiated witout te use of a starter inoculum, ave been applied in food preservation for millennia and were elucidated troug trial and error peraps over tousands of years. Te spontaneous fermentation typically results from competitive activities of varieties of contaminating microorganism (Holzapfel, 2001). Spontaneous fermentation are difficult to control, not predictable in terms of lengt of fermentation and quality of te product or product wit a sort self life and sometimes not safe since tey are liable to contamination by patogen (Novellie and de Scaeprejver, 1986; Tamime, 1990; Nout, 1992). To overcome tis problem, te most predominant microorganism found in an acceptable product are isolated and purified (Marsall, 1987; Tamime, 1990) Te medium used for te fermentation is ten pasteurized to exclude most unknown microorganism and te purified microorganisms is introduced to initiate te fermentation (Marsall, 1987; Hesseltine, 1992). By so doing te fermentation can be manipulated in suc a way tat it is possible to predict te amount and te quality of te product formed and te lengt of te fermentation period (Tamime, 1990; Hesseltine, 1992). Suc introduced cultures are termed starter culture. Process control could be acieved by environmental manipulation, wic involves controlling temperature, ph, water activity of te fermented food and may involve te use of starter organism enricment like te use of pure culture wic result to multiple starters, usually starter cultures are to be selected according to te ability to ferment sugars, rate of organic acid, optimum temperature and flavor production (Nout and Rombout, 1992). In view of te above information, tis researc terefore aimed at determining te best temperature and ph for te development of igly nutritious starter-produced weaning food. MATERIALS AND METHODS Collection of samples Brown sorgum (Sorgum bicolor) and cowpea (Vigna unguculata) used were bougt from Bodija market, Ibadan, Oyo state in a clean sterile polytene bags and kept in te refrigerator until use. Sample treatment and formulation Te collected samples were treated using te modified metod of Wakil and Onilude (2009). Te metod involved sorting, steeping, malting, oven drying and milling. Te cereal-legume formulation blends were done in a ratio of 70:30 (w/w) (Mallesi et al., 1989). Starter development Te starters used, L. plantarum and S. cerevisiae in tis work were collected from Microbial Pysiology and Biotecnology unit, Department of Microbiology, University of Ibadan, Ibadan. Nigeria based on teir recommendation by Kazeem (2009). Fermentation of te sorgum-cowpea blend Sorgum and cowpea flour (250 µm grit size) was reconstituted wit sterile distilled water at a concentration of 30% (w/v) (Livingstone et al., 1993) and inoculated wit 1ml (10 7 ) eac of te wased cells of te starters singly and mixed (in combination) culture, ten allowed to ferment for 48. Samples were taken at te initial stage (0 ) and 48 for te purpose of te analysis of te nutritional. Effect of temperature and p on nutritional Te effect of different temperatures on te nutritional of te fermenting blends was determined by incubating te Erlenmeyer flasks containing te fermenting mas in an incubator operating at 25, 30, 37 and 40 C. Te ph of te mas was adjuste d using pospate buffer and furter confirmation by potentiometric use of ph electrode. Determination of proximate composition Te metods of analysis followed were tose described by te Association of Official Analytical Cemists (AOAC, 1980). Te as was determined by incineration of known weigt of samples at 550 C until as was obtained. Protein (N x 6.25) was d etermined by te macro-kjeldal metod. Te fat composition was determined by exaustively extracting a known weigt of sample wit petroleum eter. was determined by drying te sample in te oven and cooled in desiccator overnigt at room temperature and dry matter was ten calculated (AOAC, 1980). carboydrate was calculated by difference. Determination of mineral s Te effect of starter used and fermentation on te mineral s (Mg, Zn, Fe and Ca) of te fermented blends were determined using atomic absorption spectropotometric metod as described by AOAC (1980). Determination of vitamin s Te vitamin s of te starter produced fermented weaning blends was determined using spectropotometric metod of AOAC (1980). Statistical analysis of all te data obtained was done using analysis of variance (ANOVA) and Duncan s multiple tests. RESULTS Te effect of te tree starters used (L. plantarum, S. cerevisiae and combination of te two L. Saccaromyces) on te nutritional composition of sorgum-cowpea fermented blend at 25 C is as sown on Table 1. Fermentation at 25 C increased te prot ein and te igest value (17.7%) was observed in

3 Monilola and Bukola 149 Table 1. Nutritional composition of starter-developed blends fermented at 25 C. As Proximate composition (%) Mineral (mg/100 g) Eter extract carboydrate Iron Calcium Zinc Magnesium LP a e 5.35 ab a b a LP a c 4.00 d 3.00 b a a SC a d 4.90 c 2.91 b a ab SC a cd 4.71 c 2.92 b ab b LS b b a 4.41 a b ab LS a a 5.00 bc 4.12 a b ab LP-0, Lactobacillus plantarum inoculated in unfermented gruel; LP-48 =Lactobacillus plantarum in fermented gruel after 48 ; LS-0, Lactobacillus plantarum and Saccaromyces cerevisiae inoculated in unfermented gruel; LS-48 = Lactobacillus plantarum and Saccaromyces cerevisiae in fermented gruel after 48 ; SC-0, Saccaromyces cerevisiae inoculated in unfermented gruel; SC-48, Saccaromyces cerevisiae in fermented gruel after 48. Table 2. Nutritional composition of starter-developed blends fermented at 30 C. Proximate composition (%) Mineral (mg/100 g) As Eter extract carboydrate Iron Calcium Zinc Magnesium LP d 4.30 b 5.32 a 4.30 a b LP bc bc 4.02 c 3.13 b a SC d c 4.90 b 3.91 b a SC b bc c 3.02 cd a LS c a a 4.40 a d LS a d b 3.31 b d Values are mean of tree replicates. Means tat is witin te same column wit different superscript are significantly different (p<0.05). te mixed culture L. Saccaromyces gruel wile te crude fat, crude and te carboydrate s decreased. Statistical analysis sowed tat te crude and te eter extract s of fermented gruels are significantly different from te unfermented starter developed blends. Also, fermentation of te starterdeveloped blend at 25 C increased te iron s of te blends but te increase is not significant. Values are mean of tree replicates. Means tat is witin te same column wit different superscript are significantly different (p<0.05). Te result of te gruel fermented at 30 C sows ta t fermentation increased te crude wit te igest of 19.1% recorded in te mixed culture gruel L. Saccaromyces and te least (17%) recorded in L. plantarum fermented gruel (Table 2). Fermentation decrease te moisture, crude and eter extract s of te starter-developed gruels, and tese decreases are significantly different (p<0.05) from te unfermented blends. Fermentation at 30 C a lso increases te iron and decreases te magnesium of L. plantarum fermented blends wile oter starter sowed no difference in. Generally, bot starter used and fermentation temperature does not affect te mineral statistically. Table 3 sows te effect of fermentation at 37 C temperature on te s tarter developed blends. From te table, fermentation increased te crude and total carboydrate s of L. plantarum fermented gruel, crude and as s of S. cerevisiae fermented gruel wile increase was only observed in crude of te fermented mixed culture L. Saccaromyces gruel. Te proximate composition of te fermented starterdeveloped gruel are significantly different (p<0.05) from te unfermented gruel. Te result of te analysis of te nutritional composition of te gruel fermented at 40 C sows tat fermentation increased te crude s of L. plantarum, so also te crude, as and eter extract of S. cerevisiae fermented gruel. Te mixed culture fermented gruel L. Saccaromyces ad increase in te crude and as s. Te proximate composition of te fermented starter-developed gruel are significantly different (p<0.05) from te unfermented gruel. Fermentation at 40 C decreases te mineral s of te starterdevelop blends wic were not significantly different from eac oter. Figure 1 sows te effect of different temperature (25 to 40 C) and different ph (3 to 6) ranges

4 150 Afr. J. Food Sci. Table 3. Nutritional composition of fermented starter-developed blends at 37 C. Proximate composition (%) Mineral (mg/100 g) Eter extract As carboydrate Iron Calcium Zinc Magnesium LP d a 4.30 a ab b a 0.05 b 1.40 LP b ab 3.22 c e a a 0.05 b 1.30 SC e ab 4.30 a c a a 0.05 b 1.30 SC a c 3.90 b c b b 0.05 b 1.40 LS c a 4.10 ab a b ab 0.50 a 1.40 LS a 5.11 bc 3.31 c b c ab 0.50 a 1.30 Values are mean of tree replicates. Means tat is witin te same column wit different superscript are significantly different (p<0.05). L.p-0 L.p-48 S.c-0 S.c-48 L.s-0 L.s-48 Tiamine(mg/100g) L.p-0r L.p-48r S.c-0r S.c-48r L.S-0r L.s-0 L.S-48r L.s-48 Temp ( C) A Temp (0c) ( C) B L.p-0 L.p-48 S.c-0 S.c-48 L.s-0 L.s-48 Temp ( C) C Figure 1A-1C. Effect of temperature on on vitamin of fermented starter-developed blends/gruels. on te vitamin s of te starter- developed sorgum-cowpea blends. Figure 1A sows tat riboflavin of te fermented blends increased wit increase in temperature (25 to 37 C) and later decreased at 40 C except for L. plantarum developed blend. Also, te riboflavin of all te unfermented blend is te same irrespective of te starter used wile fermented mixed culture developed blends as te igest riboflavin at all temperature used. Te analysis of te tiamine s of all te starter- developed blends sows tat 37 C is te optimum temperature for te production of te vitamins for all starters except wit L. plantarum and mixed culture blend L. Saccaromyces aving te igest tiamine (0.8 mg/100 g). Te igest of tiamine 0.5 mg/100 g was observed in te L. plantarum gruel at 40 C (Figure 1B). Te result of analysis of niacin of different starterdeveloped blends fermented at different temperatures is as sown in Figure 1C. From te figure, te igest niacin s of all te fermented starter gruel was

5 Monilola and Bukola L.p-0 Riboflavin(mg/100g) L.p-0r L.p-48r S.c-0r S.c-48r L.S-0r L.s-0 L.S-48r L.s-48 L.p-48 S.c-0 S.c-48 L.s-0 L.s-48 L.p-0 L.p-48 S.c-0 S.c-48 L.s-0 L.s-48 ph D E Niacin(mg/100g) L.p-0r L.p-48r S.c-0r S.c-48r L.S-0r L.s-0 L.S-48r L.s-48 ph F Figure 1D-F. Effect of temperature and on vitamin of fermented starter-developed blends/gruels. Table 4. Nutritional composition of te fermented starter-developed blends at 40 C. Proximate composition (%) Mineral (mg/100 g) Eter extract As carboydrate Iron Calcium Magnesium Zinc LP b 5.37 a 4.30 b 4.30 a b LP ab 4.95 c 3.96 c 2.97 d d SC c 5.31 a 5.31 a 3.94 b c SC b 4.90 c 3.92 b 3.19 c a LS bc 5.55 a 5.10 a 4.44 a 68.9 c LS a 5.10 ab 3.33 c 3.06 c a Values are mean of tree replicates. Means tat is witin te same column wit different superscript are significantly different (p<0.05). observed at 37 C except for mixed culture L. Saccaromyces starter gruel. Effect of different ph ranges on vitamin s of te fermented starter blends is as sown on Figure 1D to F. Figure 1D sows tat riboflavin of all te fermented blends increased as te ph increases (except for L. plantarum blends) from 3 to 6 wit te value ranging from 0.05 mg/100 g in fermented L. plantarum blend to 0.12 mg/100 g in fermented L. Saccaromyces blend. Te igest riboflavin in te L. plantarum blend (0.09 mg/100 g) is also observed at ph 5 and 6. Te variation in te ph from 3 to 6 does not affect te tiamine production in te unfermented L. plantarum and S. cerevisiae and fermented L. plantarum blends wile te tiamine increased wit increase ph in unfermented and fermented S. cerevisiae and L. Saccaromyces blends. At all ph, mixed culture blends ave te igest value (Figure 1E). Te result of te analysis of te niacin of te starter blends at different ph is as sown in Figure 1F, decrease in acidity/ increase in ph increases te niacin s of all te fermented starter blends wit S. cerevisiae blends aving te igest. Te result of te analysis of te effect of te variation in ph of te starter developed blends on te nutrient composition is as sown on Tables 4 to 7. Fermenting te blends at P 3 does not ave any significant effect on te

6 152 Afr. J. Food Sci. Table 5. Nutritional composition of fermenting gruel adjusted to ph 4. Proximate composition (%) Mineral (mg/100 g) Fat As carboydrate Iron Calcium Magnesium Zinc LP d b b 3.36 b 5.23 b a 1.50 b 6.30 d LP d ab b 4.10 a 5.20 b ab 1.50 b 6.40 cd SC cd 20.0 ab a 4.20 a 5.20 b bc 1.80 a 6.70 a SC bc b a 4.30 a 5.20 b bc 1.70 ab 6.60 ab LS a 20.0 ab b 3.31 b 6.60 a bc 1.51 b 6.50 bc LS a a a 3.22 b 5.41 b c 1.61 ab 6.50 bc Values are mean of tree replicates. Means tat is witin te same column wit different superscript are significantly different (p<0.05). Table 6. Nutritional composition of fermenting gruel adjusted to ph 5. Proximate composition (%) Mineral (mg/100 g) Fat As carboydrate Iron Calcium Magnesium Zinc LP c 5.20 ab 3.03 ab 5.01 a a b b LP b 5.06 b 3.10 ab 4.12 ab a ab b SC b 5.59 a 3.22 a 5.43 a b ab a SC b 5.41 ab 2.92 b 4.31 b b ab a LS b 4.02 c 3.20 ab 4.32 b a a b LS a 4.10 c 3.09 ab 4.11 bc a a b Values are mean of tree replicates. Means tat is witin te same column wit different superscript are significantly different (p<0.05). Table 7. Nutritional composition of fermenting gruel adjusted to ph 6. Proximate composition (%) Mineral (mg/100 g) Fat As carboydrate Iron Calcium Magnesium Zinc LP c 4.21 bc 3.13 b 4.21 c b b b LP b 4.65 ab 3.03 b 4.10 cd bc b b SC d 3.80 c 3.92 a 4.72 b a a a SC bc 3.96 bc 2.81 b 3.92 d b a a LS bc 3.08 c 4.02 a 5.20 a bc a b LS a 5.27 a 3.10 b 4.02 d cd b a Values are mean of tree replicates. Means tat is witin te same column wit different superscript are significantly different (p<0.05). proximate composition and te mineral s from bot witin and among te starters except crude and total carboydrate (Data not sown). Te nutrient composition of te starter-developed blends at ph 4 is as sown on Table 5. Fermented mixed culture blend as te igest crude (21.24%), moisture (90.8%), eter extract (5.41%) and crude (5.41%) s and te least, as (3.22%) and total carboydrate (64.03%). From te table, te mineral s (Fe, Ca, Mg and Zn) of bot fermented and unfermented blends were not significantly different from eac oter. Te igest crude is observed in te fermented mixed culture L. Saccaromyces blend (21.25%), and te table also sows tat adjusting te fermented gruel to ph 5 increases te crude, eter extract and te as s except for S. cerevisiae blends wile crude decreases (Table 6). Te result of te analysis of te mineral s from te table also sows tat fermentation ph and starter used as no significant effect on te blends. Table 7 sows te effect of ph6 on te nutrient composition of te tree starter-developed blends. Te result sows tat te igest crude (21.65%), eter extract (5.27%), as (4.02%), crude fiber

7 Monilola and Bukola 153 (5.20%) and te least total carboydrate (65.2%) was recorded in te fermented mixed culture blend L. Saccaromyces. Te mineral s (Fe, Ca and Zn) is te igest in te S. cerevisiae developed blend but in all, te mineral of bot fermented and unfermented blends are not significantly different (p<0.05) bot witin and among te developed blends except for Calcium and Zinc s of mixed cultured blend. Generally, at all te ph considered, te mixed cultured blends ave te igest and te least total carboydrate. DISSCUSION Tere was reduction in te moisture of te fermenting gruels for most of te temperatures and tis could be probably due to te microorganism s adsorption of water for teir microbial activities. Te significant increase in te s of te fermented gruel containing te mixed cultures L. Saccaromyces can be attributed to te proteolytic activity of te two microorganisms wic involves a symbiotic interaction wereby S. cerevisiae stimulate te L. plantarum to produce ig level of (Hamad and Fields, 1979; Zamora and Fields, 1979, Cavan and Kadam, 1989). Te s increased as te temperature increases and later decreases at 40 C.Te igest was recorded at 37 C, tis was prob ably te best optimum and favourable temperature for te growt and metabolic activities of te starters to perform teir microbial activities. Tis finding is similar to te report of te Raja et al. (2009) wo reported tat te maximum growt temperature for Lactobacillus species at temperature between 35 and 40 C. Oter researce rs (Kneifel et al., 1993; Korbekande et al., 2009; Mortazavian and Sorabvandi, 2006) also recorded tat te optimum growt temperature for most probiotics are between 37 to 40 C. Tere was reduction in te percentage of crude and carboydrate leading to te reduction of te gruel; tis may be attributed to te fact tat all te temperatures were favourable for te breakdown te starc or carboydrate to sugar in te fermenting gruel by te enzyme amylase wic ydrolysis starc granules for te growt of te organism. Tis observation is also similar to te report of Raimbault and Tewe (2001) and Ojoko (2007) wo reported te reduction of and total carboydrate in te fermentation of mango peel. For te mineral, tere was little or no increase among te four temperatures, also statistical analysis sows no significant difference (p<0.05) in te mineral of te fermented gruel developed wit L. plantarum, S. cerevisiae and te mixed cultures L. Saccaromyces, tis observation was in contrast wit te report of Oladele and Osodi (2008) wo reported tat te mineral composition increased wit fermentation time, but te findings of Salin (1999) was different, wo notice tat mineral is not affected by fermentation unless some salts are added to te product during fermentation. Tere was increase in te vitamin s of te fermenting gruel generally; te igest values were recorded in te mixed cultures at 37 C and wic may be due to te fact t at te mixed starter cultures consisting of lactic acid bacteria and fermenting yeast are important in acieving biological stability by assimilating all available polymer to produce simple more nutrients suc as vitamins. An observation similar to tat of Kneifel and Mayer (1991) wo reported tat te two cultures sows 40% increase in tiamine s and riboflavin of kefir. Tese was also similar to Sanni et al. (1999) reports were S. cerevisiae and L. plantarum were used as starter cultures to ferment various cereals in te production of weaning foods, an increase in te s of riboflavin, tiamine, niacin and ascorbic acid was reported during fermentation. Te result of te analysis of te effect of ph on te starter produced gruel sows tat te of te mixed culture L. Saccaromyces fermenting gruel as te igest at ph 6. Te increase in te may be as a result of te microorganism present in te gruel wic breaks down complex to amino acid at optimum ph 6, wic it seems to be favourable for most lactic acid bacteria and yeast, an observation similar to tat of Loubiere et al. (1997). Furtermore, mixed culture of L. plantarum and S. cerevisiae ad te igest wic may be due to te fact tat yeast stimulates te growt of te lactic acid bacteria wic is already known tat activity increases wile cells are growing exponentially and tis correlate wit te report of Ketarpaul and Cauan (1990) wo found out tat mixed cultures of tese organisms during te fermentation of pearl millet flour improves its biological utilization in rats. Tere was reduction in te carboydrate and te at all te ph and all te starter culture combination used. Tis may probably due to te microbial activities of te organisms, as a result of utilization of te available starc for metabolic activities for te growt of te microorganism, an observation similar to te work of Ejiofor and Okafor (1981) wo confirmed te reduction of carboydrate by te activities of amylase for te breakdown of starc to simple sugars. Te effect of ph on te mineral of te fermenting gruel was little or not significantly different (p< 0.05) at all te different phs, tis may be due to te fact tat te organisms migt convert some compound of te fermenting gruel to a useable minerals. Tis observation was also reported by Salin (1999) wo notice tat mineral is not affected by fermentation and Betscart (1988) also confirm te same result. Te analysis of te fermenting gruel at different phs sows tat te igest vitamin was recorded at ph6 of te fermenting gruel wile te ph 3 sows no significant

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