Int.J.Curr.Microbiol.App.Sci (2014) 3(3): 888-895

ISSN: 2319-7706 Volume 3 Number 3 (2014) pp. 888-895 http://www.ijcmas.com

Original Research Article Microbiological quality of fermented (Gari) sold in Ota Ogun State

B.K.Olopade, S.Oranusi*, R.Ajala, and S.J.Olorunsola

Biological Sciences Department, Covenant University,Ota, Ogun State, Nigeria *Corresponding author

A B S T R A C T

Thirty six gari samples (eighteen each of white and yellow types) were subjected to microbial analysis. Samples were serially diluted to 104 and appropriate dilutions inoculated by spread plate method onto Nutrient agar, MacConkey agar and Potato- K e y w o r d s Dextrose agar plates for Total aerobic plate count (TAPC), Coliform count (CC) and Fungal count respectively. TAPC for white gari ranged from 2.0x102 to Gari samples; 1.1x104, coliform count ranged from no growth (NG) to 7.1x103 while fungal count Total aerobic ranged from no growth to 6.0x102. The microbial load of yellow gari ranged from plate count; 1.0x102 to 5.0x103 for TAPC, NG to 6.0x103 for coliform count and NG to 3.0x103 coliform for fungal count. The bacteria isolates from the various samples include Bacillus count; spp Enterobacter spp, Pseudomonas, Staphylococcus and Klebsiella spp. Fungi moisture isolated includes Aspergillus niger, Aspergillus fumigatus, Fusarium, Rhizopus and content. Penicillium spp. The pH of the samples ranged from 4.76 to 4.94 in the yellow type and 4.78 to 4.91 in the white type. The moisture content was 6 to 8 percent in yellow type and 4 to 7 percent in the white type. Application of good manufacturing practices (GMP) and HACCP in gari production is imperative.

Introduction

Cassava roots contain a fibrous peel (10- biochemical contents of such products are 15% of tuber weight) and a core, the main affected (Kemdirim et al., 1995). region for starch (IITA, 1990). Cassava can be consumed in various forms: boiled, Amongst the various fermented cassava baked or fried and it is processed before products, Gari is the most commonly consumption in order to detoxify and consumed in Nigeria and accounts for preserve it (Oyewole and Sanni, 1995; 70% of the entire cassava production in Obadina et al., 2009). The main cassava Nigeria (IITA, 1990). Several millions of food products of considerable domestic people in the African continent consume importance in Nigeria are gari, lafun and Gari and it forms a significant part of the . When cassava roots are processed people s diet especially in the West into products such as gari and flour, the African sub region (Edem et al., 2001;

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Ogiehor et al., 2007 and Kostinek et al., fermented pulp to about 10% moisture 2005). content and may result in partial dextrinization of starch. Asegbeloyin and Gari is a granulated and dehydrated, Onyimonyi, (2007); Harbor and cassava product. It is classified/ grouped Ogundu(2009) also revealed that frying based on texture, length of fermentation, destroys enzymes and microorganisms and region or place where it is produced and aids in eliminating cyanide gas from the colour imparted by the addition/non- product. addition of palm oil. It has a high swelling capability and can absorb up to four times There are reports on the high load of its volume in water. (Jekayinfa and microorganisms in gari sold in the market Olajide, 2007).Obtainable in the market is (Ogiehor et al., 2007; Ijabadeniyi, 2007; the dry form of post processed gari which Amadi and Adebola, 2008). The can be consumed soaked in cold water. microorganisms reported for market Sugar can also be added to the soaked gari samples include:Salmonella spp., and it can be eaten with meat, roasted Klebsiellaspp.,Pseudomonas spp.,Bacillus groundnuts, smoked fish, boiled beans, spp., Clostridium spp.,Fusariumspp., coconut, palm kernel, groundnut cake- Aspergillusspp; Penicilliumspp.,Rhizopus kwuli kwuli, and fermented snacks spp and Cladosporiumspp. There may be . Beverages and milk may also be economic losses andfood borne illnesses added as complements. is another as a result of contamination by these food prepared from gari. The granules are microorganisms. This study was carried added into hot water and stirred to form a out to assess the microbial quality of gari stiff paste which can be eaten with sold in Ota, with a view to educating the indigenous or (Asegbeloyin public on the need for food safety and Onyimonyi, 2007). consciousness and contributing to the body of knowledge on this all important staple Gari production is a tasking and African food. burdensome procedure and its method of production differs from one locality to Materials and Methods another. In a typical production of Gari, the cassava tubers are peeled, washed, Collection of samples grated and packed into closely woven bags. The poisonous juice can then be The study was carried out between March removed by placing a heavy object on the and May, 2013. A total of thirty six (36) bag and the contents of the bag are samples of gari were purchased from three allowed to undergo spontaneous solid state major markets in Ota - Sango Ota, Oju- fermentation for several days at ambient Ore and Oja-Ota. Six(6) each of yellow temperatures (Ray and Sivakumar, 2009; and white gari types were randomly Huch et al., 2008). According to purchased from each market in the order Akindahunsi et al.(1999), Azam- Ali et al. of duplicate samples from three different (2003) the grated tubers are allowed to but major gari food vendors per market. ferment in order to preserve the product, The samples were appropriately labeled to reduce cyanide and enhance its flavor. indicate the name of the market, gari type Osho and Dashiell (2002) stated that (white or yellow), sample number, date frying at high temperatures dries the and time of collection. Samples were

889 Int.J.Curr.Microbiol.App.Sci (2014) 3(3): 888-895 transported in sterile polyethylene bags to (EMB) agar for confirmatory test. Plates the laboratory for analysis within one hour were incubated for 24h at 37°C and 44°C of collection. respectively. Growth of characteristic colonies on EMB constitute confirmatory Sample Analysis test positive. Colonies from confirmatory test were Gram stained and inoculated into Ten gram (10g) samples of gari were lactose broth for completed coliform test. homogenized in 90 ml sterile distilled Gas production and/or colour change of water (10-1 dilution), further serial dilution dye plus Gram negative non-spore bearing of sample homogenate to 10-4 was carried rod represent presence of coliform (Speck, out also in sterile distilled water. 1976 Oranusi et al., 2004). Absence of Approximate 0.1ml aliquot of appropriate growth was recorded at 44°C incubation dilutions were spread plated on plates of for all the samples indicating absence of Nutrient agar, MacConkey agar and Potato fecal coliforms. Dextrose agar (all from Biolab, Hungary) for total aerobic plate count, coliform Enumeration and identification of count and fungal counts respectively. microorganisms isolated Sample homogenates were also inoculated onto Manithol Salt agar (Biolab, Colony counts at the expiration of Hungary), Salmonella-Shigella agar incubation time was with digital colony (Oxoid, England) after pre-enrichment in counter (Gallenkamp, England), total Selenite F broth, for isolation of microbial population was expressed as staphylococci and salmonellae. One gram colony forming units per gram (cfug-1) of samples were inoculated into Lactose sample. Pure cultures of isolates were broth (Biomark, India) in screw capped obtained by repeated subculture on test tubes with inverted Durham tubes for nutrient agar and were stored on slants at coliform test. All culture plates were 4°C until characterized. Characteristic incubated at 37°C aerobically for 24-48h. bacteria isolates were identified based on Potato Dextrose agar (PDA) plates for colonial morphology, microscopy and fungal culture was however, incubated for biochemical tests (Holt et al., 1994). 72-120 h at laboratory room temperature Identification of Fungal isolates was based of 29±2°C. Culture plates were examined on morphological characteristics and for enumeration and identification of microscopy with reference to standard colonies at the expiration of incubation atlas and keys (Samson and Reenen- period Hoekstra, 1988; Tsuneo, 2010).

Coliform Test Determination of pH and moisture content of gari samples Aliquot 1g samples in test tubes with inverted Durham tubes in Lactose broth The pH of the samples was determined were incubated at 37°C for 24-48h. Tubes following the method described by showing gas production and/or colour Ogiehor and Ikenebomeh (2005). Ten change of dye were reported as grams of each sample were homogenized presumptive coliform test positive. These in 10ml of distilled water and the pH of positive tubes were streaked out on the suspension determined using a duplicate plates of Eosin Methylene Blue reference glass electrode pH meter

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(Mettler Delta 340, Mettler tocedo thereafter sieved. Products are often limited. UK). The moisture content was displayed in open basins/bowls for sales in determined by drying 2g sample at 100- the market place. The presence of coliform 105°C to constant weight (AOAC, 1990) could therefore be from post process contamination via food handlers and the Results and Discussion environment. The ICMSF (1996) and the African Organization for Standardization Table 1 shows the mean microbial count recommended absence of coliform in of white and yellow types of gari samples ready to eat foods. The presence of obtained from three major markets in Ota, coliform in most of the gari samples Ogun state Nigeria. It reveal that most of therefore makes it of poor quality for the samples had TAPC within the limits of human consumption. 101 to 103cfug-1and are contaminated with coliforms and fungi to about the same The fermentation of gari is by mixed order. The table also shows that only two microbial cultures, this could have samples of white gari had TAPC count to accounted for the diverse microbial the order of 104cfug-1. The pH of the population contaminating the product. samples is of acidic range 4.76 to 4.94 in Similarly post process contamination both the white and yellow types of gari. specifically associated with sieving of Table 2 reveals that the gari samples were products after heat treatment and the contaminated by diverse microbial spp spreading of products in the open to air mainly of Bacillus, Pseudomonas, dry, coupled with the practice of leaving Klebsiella, Staphylococci and Moulds. gari open for sales could have accounted The percentage moisture contents of the for the diverse microbial population. The samples range from 4 to 7 in the white gari isolation of diverse microbial species from and 6 to 8 in the yellow types. this ready to eat food (gari) corroborate the findings of Nichols et al. (1999), Mensah The gari samples contain total aerobic et al. (2002), Idowu (2006), Taulo et al. plate count and fungi counts within (2008), Oranusi and Braide (2012), acceptable limit. Ready to eat foods with Oranusi et al.,2013). The use of specific plate counts of 103 are acceptable, counts starter culture, effective HACCP of 104 to 105 are tolerable while counts application and good manufacturing 106 are unacceptable (ICMSF, 1996). The practice from farm to fork will help curtail presence of contaminants to the order of the level of contamination. Although the 104 in two of the gari samples however, microbial load counts with the exception negates the 103 stipulated requirements by of coliform counts are within acceptable the African Organization for standard limits (ICMSF, 1996), the Standardization. Coliform was detected in presence of B. cereus and S. aureus calls most of the gari samples at high counts 102 for concern because some strains of these to 103, although no fecal coliform was organisms are known to be toxigenic and detected (absence of growth of coliform have been implicated in food borne organisms at 44°C), the presence of intoxication (Mensah et al., 1999; Oranusi coliforms generally signifies poor sanitary et al., 2007), their presence therefore calls condition in the production of gari. Gari for concern. B. cereus is common after frying (heat treatment) is often spread environmental contaminants while S. in the open to cool and air dry and it is aureus is of human origin, their presence

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Table.1 Mean microbial load cfug-1 and pH of gari

Sample White gari Yellow gari outlet TAPC Coliform Fungi pH TAPC Coliform Fungi pH count count count count Sango- 1 4.7 × 103 2.0 × 103 4.0 × 102 4.81 1.5 × 103 1.1 × 103 8.0 × 102 4.79 Ota market 2 3.1 × 103 3.0 × 103 3.0 × 102 4.90 2.8 × 103 1.0 × 103 3.0 × 102 4.88

3 5.6 × 103 2.7 × 103 3.0 × 102 4.87 1.0 × 103 1.4 × 103 4.0 × 102 4.84

4 4.7 × 103 3.0 × 102 4.0 × 102 4.82 2.1 × 102 4.0 × 103 7.0 × 102 4.80

5 2.0 × 102 NG 4.0 × 102 4.78 1.1 × 102 1.4 × 102 2.0 × 102 4.89

6 1.1 × 104 2.7 × 103 3.0 × 102 4.81 4.3 × 103 3.4 × 103 5.0 × 102 4.85

Ojuore 1 2.0 × 102 2.0 × 102 3.0 × 102 4.91 3.0 × 102 6.0 × 103 1.0 × 103 4.92 market 2 2.0 × 102 2.0 × 102 6.0 × 102 4.89 4.0 × 102 1.0 × 102 4.0 × 102 4.86

3 4.0 × 102 NG NG 4.89 2.0 × 102 8.0 × 102 6.0 × 102 4.92

4 3.1 × 103 7.0 × 103 4.0 × 102 4.80 1.6 × 103 2.0 × 103 9.0 × 102 4.93

5 2.0 × 102 2.0 × 102 3.0 × 102 4.89 2.9 × 103 2.0 × 102 4.0 × 102 4.87

6 9.6 × 103 3.0 × 103 4.0 × 102 4.86 1.1 × 103 9.0 × 102 5.0 × 102 4.94

Ojaota 1 1.0 × 104 8.0 × 102 4.0 × 102 4.83 1.0 ×102 NG NG 4.80 market 2 9.6 × 103 7.1 × 103 3.0 × 102 4.79 1.0 × 102 1.0 × 102 1.0 × 102 4.78

3 4.0 × 102 3.0 × 102 1.0 × 102 4.82 1.9 × 103 4.2 × 103 4.0 × 102 4.84

4 8.0 × 102 NG 6.0 × 102 4.90 5.0 × 103 3.1 × 103 3.0 × 103 4.78

5 3.0 × 102 2.0 × 102 1.0 × 102 4.88 4.1 × 102 1.2 × 102 5.0 × 102 4.76

6 5.6 × 103 2.0 × 103 3.0 × 102 4.88 6.0 × 102 NG 1.2 × 102 4.82

Key: NG= no growth; TAPC= total aerobic plate count

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Table.2 Microorganisms isolated from gari

Sample Microorganisms isolated White gari Klebsiella spp, Bacillus licheniformis, Pseudomonas aeruginosa, Lactobacillus spp, Staphylococcus aureus, Staphylococcus epidermidis, Penicillium spp, Aspergillus niger, Rhizopus spp Yellow gari Bacillus spp, Enterobacter spp, Pseudomonas, Bacillus cereus, Staphylococcus aureus, Bacillus megaterium, Klebsiella spp, Fusarium spp, Mucor spp, Aspergillus niger, Aspergillus fumigatus could therefore be from the food handlers, basins/ bowls. Optimization of the process utensils and the environment. The to using a starter culture with reduced post processing of gari for consumption often process handling is imperative. Effective involve mild or no heat treatment, the HACCP and GMP will help reduce or toxins from B. cereus and S. aureus if and eliminate product contamination and thus when elaborated in food are heat stable, make the product safe for consumption. the consumption of contaminated gari could therefore portend a potential risk to References consumers. Akindahunsi, A.A., Oboh, G. and Oshodi, Moulds are common environmental A.A. 1999. Effect of fermenting contaminants due to their ability to cassava with Rhizopusoryzae on the produce spores; this could explain their chemical composition of its flour and presence in gari. They have been . Riv. Ital. DelleSost. Grasse, 76: implicated in ready to eat foods and in 437-440. unregulated/ mixed fermentation. Species Amadi, J.E. and Adebola, M. O. of Aspergillus, Penicillium and Fusarium 2008.Effect of moisture content and are known to produce deleterious storage conditions on the storability of mycotoxins under favourable conditions garri.African Journal of (Sweeney and Dobson, 1998; Kabak et al., Biotechnology.724: 4591- 4594. 2006; Oranusi et al., 2013), their presence AOAC 1990 Official Method of Analysis, in gari must therefore be treated with 13th ed., The Associ-ation of caution. Analytical Chemists. Asegbeloyin, J.N. and Onyimonyi, A.E. Gari is a basic staple food in Nigeria and 2007.The effect of different processing some African countries, it accounts for methods on the residual cyanide of 70% of the entire cassava production in Garri .Pakistan. Journal of. Nutrition., Nigeria (IITA, 1990). There is therefore a 62: 163-166. need to maintain proper sanitary Azam -Ali, S., Judge, E., Fellows, P. and conditions so as to avoid health risks. The Battcock, M. 2003. Small-scale food moisture content of gari samples analyzed processing. A directory of equipment is low and within standard specification, and methods.2nd edition.ITDG this could have accounted for keeping the Publishing.pp: 256 microbial load of gari low. It is therefore Edem, D. O., Ayatse, J.O.I. and Itam E.H. recommended that gari should be sold well 2001. Effect of soy protein packaged in bags and not as exposed in supplementation on the nutritive value

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