STUDIES ON FERMENTATION OF CASSAVA VARIETIES BY DIFFERENT SPECIES OF LACTIC ACID BACTERIA AND EFFECT ON CHEMICAL, FUNCTIONAL AND SENSORY PROPERTIES OF FUFU FLOURS PRODUCED

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ABSTRACT


Cassava fufu flours were produced following 72 h fermentation, of two cassava varieties (TME 419 and Umucass 45) by lactic acid fermentation and natural fermentation (0-72 h). Two starter cultures (L. plantarum, L. fermentum and their combination) were used to initiate lactic acid fermentation. At 24 h intervals the pH, titratable acidity (TTA), microbial load (Starter culture count, total LAB count of natural fermenting mash, yeast/mould count, microbial quality) and molecular characterization of natural fermentation isolates were determined. The wet mash samples were dried and milled to flour and analyzed for chemical, functional and sensory properties of the flours. The results revealed that the percentage titratable acidity of the cassava varieties increased from 0.23 – 1.18% as fermentation progressed in relation to decrease in pH (5.22 -3.70). Samples (419 LP+LF and 45LP) gave the highest starter culture count of 5.62 log CFU/g at 48 h and showed a decrease at 72 h of fermentation. The sequencing of PCR of 16s rRNA gene, identified the bacterial isolates as Lactobacillus plantarum, Lactobacillus fermentum, Lactobacillus pentosus, Lactococcus lactis and Saccharomyces cerevisae. The cassava flour samples were safe for consumption since potential pathogens were not detected. The fresh cassava carotenoid content of Umucass 45 had the highest carotenoid value (11.7mg/g) which showed significant difference (p < 0.05) from other samples. The sugar and starch contents showed similar trends. Sugar content ranged from (4.93 – 2.20), with unfermented sample having the highest value. The proximate composition of the products ranged from; Moisture (8.58 – 11.14%), Ash (1.30 – 2.54%) crude protein (1.38 – 5.73%), crude fibre (0.70 – 1.76%), fat (0.52 – 0.93%) and carbohydrate (81.09 – 84.84%).  Functional properties increased in bulk density (0.66 – 0.79 g/ml), oil absorption (1.25 – 1.36 g/ml), water absorption (2.98 – 3.52 g/ml), swell index (2.73 – 3.45 g/ml), while a decrease in gelatinization temperature (70.10 – 67.450c) was observed. Fermentation reduced significantly, the anti-nutritional factors: HCN (48.03 - 3.16mg/100g), phytate (5.12 – 0.23 mg/100g), tannin (2.22 – 0.20 mg/100g), alkaloid (3.84 – 0.06 mg/100g). Amino acid profile revealed that fermentation led to significant improvement of   protein quality of the flour products. The highest essential amino acid leucine (6.36 g/100g) and non-essential amino acids glutamic acid (7.64 g/100g) were recorded for flour from yellow cassava. Minerals of samples ranged from 28.42 – 31.10 mg/100g for calcium; 15.30 – 21.67 mg/100g magnesium; 35.29 – 46.31 mg/100g potassium; 13.07 – 18.33 mg/100g phosphorus; 0.67 – 1.75 mg/100g zinc; and 1.25 – 3.52 mg/100g iron respectively. The Gas Chromatograph Mass Spectrometer (GCMS) profile of samples detected the presence of alcohol (penatdecanol) and acids (hexadecanoic acid) indicating the flour products contain different flavor compounds, due to different microorganisms present during processing. Sensory evaluation, showed that starter culture and natural fermented flours were not significantly different (p > 0.05) in odour mould-ability, hand feel and overall acceptability but showed significant difference (P < 0.05) in appearance and acceptable. The flours had good functional and sensory properties. Fermentation either natural or with starter cultures can enhance the nutritive value of cassava tubers for sustainable local technologies and utilization of the value-added product for human nutrition.





TABLE OF CONTENTS

Title Page                                                                                                                    i

Declaration                                                                                                                  ii

Certification                                                                                                                iii

Dedication                                                                                                                  iv

Acknowledgments                                                                                                      v

Table of Contents                                                                                                       vi

List of Tables                                                                                                              xi

List of Figures                                                                                                             xiii

List of Plates                                                                                                               xiv                    

Abstract                                                                                                                      xv

 

CHAPTER 1:            INTRODUCTION                                                                           1

1.1       Statement of Problem                                                                                                 6

1.2       Justification                                                                                                     7

1.3       Objectives of The Study                                                                                 8                                                                                                         

CHAPTER 2:            LITERATURE REVIEW                                                              9

2.1       Fermentation                                                                                                   9

2.2       Fermented Foods                                                                                            11

2.3       Types of Food Fermentation                                                                          11

2.3.1    Fermentation with lactic and bacteria (LAB)                                                 11

2.3.2    Acetic acid fermentation                                                                                12

2.3.3    Alcoholic fermentation                                                                                   13

2.3.4    Traditional fermentation                                                                                 13

2.4       Lactic Acid Bacteria (LAB) Metabolism of Carbohydrates                          15

2.5       Important Characteristics of Lactic Acid Bacteria                                         16

2.6       Importance of Fermented Foods                                                                    17

2.6.1    Healthy and safe products                                                                              17

2.6.2    Probiotic and prebiotic potentials                                                                   17

2.6.3    Nutritional and health benefits of LAB fermentation                                    18

2.7       Nigeria Indigenous Fermented Foods (NIFF)                                                18

2.8       Advantages of Fermented Foods                                                                   21

2.8.1    Role and function of fermentation on cassava food                                      22

2.9       Anti-nutrient Reduction in Cassava Fermented Food                                                23

 

 

CHAPTER 3:            MATERIALS AND METHODS                                                   25

3.1       Location                                                                                                          25

3.2       Materials                                                                                                         25

3.3       Organisms and Culture Methods                                                                    25

3.4       Preparation of Inoculum                                                                                 26   

3.5       Preparation of Fermented Yellow Cassava Roots                                          26

3.6       Yellow Cassava Fufu Flour Production                                                          27

3.7       Determination of Titratable Acidity (%)                                                        33

3.8       pH Measurement                                                                                             33

3.9       Isolation and Enumeration of Lactic Acid Bacteria (Lab) and Yeast

            Population During Natural Fermentation of the Cassava Varieties                34

3.9.1    Yeast count and isolation                                                                               35

3.9.2    Microscopic characterization                                                                          35

3.9.3    Gram staining                                                                                                  35

3.9.4    Motility test                                                                                                    36

3.10     Biochemical Tests                                                                                           36

3.10.1  Catalase test                                                                                                    36

3.10.2  Oxidase test                                                                                                    36

 

3.11     DNA Extraction of Lactic Acid Bacteria (Lab) and Yeast Isolated

            During Natural Fermentation of Cassava Varieties.                                       37

3.11.1  DNA extraction                                                                                              37

3.11.2  Polymerase chain reaction (PCR) amplification of extracted DNA               38

3.11.3  The PCR purification                                                                                      39

3.11.4  DNA sequencing and BLAST search                                                             40

3.12     Total Carotenoids Determination                                                                   40

3.13     Determination of Starch and Sugar Contents                                                             41

3.14     Determination of Proximate Composition of Yellow Cassava Flours            42

3.14.1  Moisture determination                                                                                   42

3.14.2  Carbohydrate content determination                                                              43

3.14.3  Ash content determination                                                                             43

3.14.4  Fat determination                                                                                            43

3.14.5  Determination of crude fibre                                                                          44

3.14.6  Determination of protein                                                                                45

3.15     Functional Properties Determination                                                              45

3.15.1  Bilk density                                                                                                     45

3.15.2  Gelatinization temperature (GT)                                                                     46

3.15.3  Water absorption capacity (WAC)                                                                 46

3.15.4  Oil absorption capacity (OAC)                                                                       46

3.15.5  Swelling index                                                                                                47

3.16     Anti-nutrient Composition                                                                              47

3.16.1  Determination of alkaloid                                                                               47

3.16.2  Determination of tannins                                                                                48

3.16.3  Determination of phytates                                                                              48

3.16.4  Determination of hydrogen cyanide                                                               48

3.16.5  Determination of amino acid profile                                                               49

3.17     Defatting Sample                                                                                            49

3.18     Nitrogen Determination                                                                                  49

3.19     Hydrolysis of the Sample                                                                               50

3.19.1  Loading of the hydolysate into analyzer                                                        51

3.19.2  Method of calculating amino acid values                                                       51

3.20     Determination of Minerals                                                                              51

3.20.1  Determination of calcium and magnesium                                                     51

3.20.2  Determination of potassium                                                                            52

3.20.3  Determination of phosphorus                                                                         52

3.20.4  Determination of iron and zinc                                                                       53

3.21     Identification and Qualification of Flavor Profile Fermented Flours            54

3.22     Sensory Evaluation of Fufu Flour                                                                  54

3.23     Statistical Analysis                                                                                          55

 

CHAPTER 4:            RESULTS AND DISCUSSION                                                     56

4.1       pH and Titratable Acidity of Fermented Cassava Varieties                           56

4.2       Total Lactic Acid Bacteria Count                                                                   59

4.3       Lactic Acid Bacteria Profile During the Natural Fermentation of

Cassava Varieties                                                                                            62

 

4.4       Molecular Identification of Isolates During Natural Fermentation               64

4.5       Yeast Count (Log cfu/g) During the Natural Fermentation of the

Cassava Varieties.                                                                                           78

4.6       Microbial Count (cfu/g) of the Fermented Cassava Flour Samples                81

4.7       Carotenoid Content of the Cassava Flours                                                     83

4.8       Changes in Starch and Sugar Content of the Cassava Flours                        86

4.9       Proximate Composition of Raw and Fermented Cassava Flours                   88

4.10     Functional Properties of Fermented Cassava Flours                                      94

4.11     Amino Acid Profile of Fermented Cassava Flours.                                        98

4.12     Anti-Nutrient Content of Fermented Cassava Flours                                    99

4.13     Mineral Content of the Fermented Cassava Flours.                                       102

4.14-21 Flavor Profile of Fermented Cassava Flour.                                                  106

4.15     Sensory Properties of Fermented Cassava Fufu Flour                                    117                                       

CHAPTER 5: CONCLUSION AND RECOMMENDATIONS                         119

5.1       Conclusion                                                                                                      119

5.2       Recommendations                                                                                          121

References                                                                                                      122

Appendix                                                                                                        140






LIST OF TABLES

3.1                   PCR Conditions for 16s rRNA of bacteria                                        38

 

3.2                   PCR conditions for its fungi                                                               38

 

3.3                   PCR cocktail mix                                                                                39

 

4.1:                  The pH and Titratable Acidity of starter fermented cassava

                        varieties at different fermentation periods                                         58

 

4.2.                  Lactic acid bacteria (LAB) count (1ogCFU/g) during

                        fermentation periods with starter cultures of

L. plantarum, L. fermentum and both                                                 61

 

4.3:                  Lactic acid bacteria (LAB) profile during natural

                        fermentation of cassava varieties (log CFU/g)                                  63

 

4.5:                  Yeast/mould count (log CFU/g) during natural fermentation of

                        cassava varieties                                                                                  80

 

4.6:                  Microbial count (CFU/g) of fermented cassava flours.                      82

 

4.7                   Effect of starter cultures on the carotenoid retention of

cassava flours                                                                                      85

 

4.8:                  Effect of starter culture fermentation on starch and sugar

                        content of flour samples                                                                     87

 

4.9:                  Effect of starter culture fermentation on the proximate

                        composition (dry weight basis) of cassava flour samples                   93

 

4.10:                Effect of starter culture fermentation on the functional

                        properties of cassava flour samples                                                     97

 

4.11:                The amino acid profile (g/100g protein) of fermented cassava

fufu flours of TME 419 and Umucass 45 cassava varieties                99

 

4.12:                Effect of starter fermentation on anti-nutrient produced

fufu flours                                                                                            102

 

4.13:                Effect of starter culture fermentation on the mineral content

                        (mg/100g) of the fermented flours                                                     106

 

4.14:                GCMS retention time and area percentage (concentration)

            .           of flavor compounds in natural fermented cassava flour (NF 419)    110                                                                 

4.15:                GCMS retention time and Area % (concentration) of flavor

compounds in starter culture fermented cassava flour (LP 419)        111

 

 

4.16:                GCMS retention time and area % (concentration) of flavor

compounds in starter culture fermented cassava flour (LF 419)        112

 

4.17:                GCMS retention time and area % (concentration) of flavor

compounds in starter culture fermented cassava flour (Both 419)     113

.

4.18:                GCMS retention time and area % (concentration) of flavor

compounds in natural fermented cassava flour (NF 45)                     114

 

4.19:                GCMS retention time and area % (concentration) of flavor

compounds in starter culture fermented cassava flour (LP 45)          115

 

4.20:                GCMS retention time and area % (concentration) of flavor

compounds in starter culture fermented cassava flour (LF 45)          116

 

4.21:                GCMS retention time and area % (concentration) of flavor

compounds in starter culture fermented cassava flour (both 45)        117

 

4.22:                Effect of starter culture fermentation on the sensory properties

                        of instant cooked fufu flour                                                                119

 






LIST OF FIGURES


3.1:                  Traditional method of production of cassava flour                            28

                                               

3.2:                  Traditional method of production of cassava flour with

                        inoculation of Lactobacillus plantarum                                              29

 

 

3.3:                  Traditional method of production of cassava flour with

                        inoculation of L. fermentum                                                                30

 

 

3.4:                  Traditional method of production of cassava flour with

                        inoculation of L. plantarum and L. fermentum                                   31

 

 

3.5:                  Traditional method of production of unfermented cassava flour       32

 

4.1:                  L. Plantarum  (NF 45 MRS)                                                               70

 

 

4.2:                  L. Fermentum (NF45 MRS)                                                               71

 

4.3:                  L.Pentosus (NF45 MRS)                                                                    72

 

 

4.4:                  S. Cerevisiae (NF45 PDA)                                                                 73

 

 

4.5:                  L. Fermentum (NF 419 MRS)                                                                        74

 

 

4.6:                  L. plantarum (NF419 MRS)                                                               75

 

 

4.7:                  L. Lactis (NF419 MRS)                                                                      76

 

 

4.8:                  S. cerevisiae (NF419 PDA)                                                                77

 

 

 

4.9:                  S. cerevisiae (NF36 PDA)                                                                  77

 

 

 

 

 

 

 

 

 

 

 

LIST OF PLATES

 

1:         Shows the total genomic DNA of L. Plantarum, L. fermentum and

            L. pentosus as shown in agarose gel electrophoresis for 45 NF MRS.           65                                          

 

 

2:         Total genomic DNA of L. plantarum, L. fermentum and L. lactic as

            revealed in agarose gel electrophoresis for NF 419 MRS                               65

 

 

3:         Shows the total genomic DNA of  S. cerevisiae as revealed in

agarose gel electrophoresis for NF 45 PDA.                                                  66

 

 

4:          Total genomic DNA of  S.cerevisiae as shown in agarose gel

             electrophoresis for NF 419  potato dextrose agar (PDA)                              66

 

5:         Shows the PCR products of L. plantarum, L. fermentum, L. pentosus

for NF 45                                                                                                        68

 

6:         Shows the PCR products of L. plantarum, L. fermentum, L. pentosus

for NF 45                                                                                                        68

 

7:         Shows the PCR product of  S. cerevisiae for NF 45 PDA (PCR

reaction gel picture of fungi from NF45)                                                       69

 

8:         Shows the PCR product of S.cerevisiae for NF 419 PDA                             69

 

 

 

           



 

 

CHAPTER 1

INTRODUCTION

 

Cassava (Manihot esculenta Crantz) is an important crop widely cultivated in Sub-Saharan African. Although cassava is grown virtually in all parts of the sub-continent, its production is specific in the humid tropics (Okereke, 2001). Cassava is one of the major essential food crops in the tropic (Burrel, 2003) and used as a food preservation and income generation crop for many millions of people in the unindustrialized world (Scott et al., 2002).

 

Cassava is grown widely in Nigeria and in many regions of the tropics, where it serves as one of the basic food sources for about 200-300 million people (FAO, 1991). Cassava is a major starch staple in Africa and it is particularly important in Nigeria. Nigeria accounts for about 40% of cassava production in Africa. According to Oyewole and Odunfa (1992) in African, cassava provides over 50% of the average daily caloric intake in some countries. A diversity of national efforts to propagate optimized cassava cultivars have recently been launched (Guira et al., 2016) to increase yield and opposition to infections and through Harvest Plus Germplasm Development - African component develop cassava cultivars with increases levels of β-carotene in the root tubers. Cassava cultivars rich in pro-vitamin A (PVA) have been released in cassava-growing countries in Africa as a consequence of effective biofortification attempts across the world (Omodamiro et al., 2012; Birol et al., 2015). The new varieties besides adding to the energy intake of consumers also acts as vehicle of conveying Pro-vitamin A (PVA) to vitamin A deficient (VAD) populations (Tanumihardjo et al., 2008). Yellow cassava is another name for these new bio-fortified cassava roots. The yellow fleshed cassava according to (Egesi et al., 2011) is a newly released bio-fortified crop which is similar to the white varieties in terms of utilization for man and animal, though the pulp colour differs. The yellow-fleshed cassava varieties are grown in Nigeria for their high accumulation of β-carotene, a precursor of vitamin A. Yellow-fleshed cassava shows great potential to alleviate vitamin A deficiency in Africa since cassava is a staple food. Vitamin A is also a vital part of human nutrition as it helps with sight, cell division, glycoprotein formation, fertility, and total development and growth (Woolfe, 1992). African countries are not only faced with problem of food preservation but that of nutritional insecurity leading to different forms of micronutrients deficiencies in the diet. Bio-fortification of cassava is therefore highly appropriate as this will contribute to the alleviation of diseases associated with vitamin A deficit (VAD) which is a common dietary health problem, especially in countries where cassava is a major staple food.

Therefore, its development and dissemination by National Root Crops Research Institute Umudike (NRCRI) and International Institute of Tropical Agriculture (IITA) Ibadan in collaboration will compliment current effort to address vitamin A deficiency by delivering vitamin A through a staple food consumers eat every day (www.harvestplus.org). Nigeria is the major producer of cassava in the world (FAO, 2008) with about 45 million metric tonnes and cassava transformation is the most advanced in Africa (Egesi et al., 2006).

Cassava is cultivated in the tropical regions and may be the main valuable root crop in terms of overall output and cultivated area (Ano, 2003). According to Ogbe et al. (2007), it is a main food crop in Nigeria. It is strategically important for its position in food protection, alleviating poverty, and input materials supplies for agro-allied sectors in Nigeria as well as its export market potential (Egesi et al., 2007). In municipal regions, cassava consumption of poor households is double that of non-poor households. In rural area, poor households intake of cassava is triple that of non-poor households. When dry, cassava is storable and moveable across greater routes. The evidence from the Collaborative Study of Cassava in Africa shows that cassava can be converted into a broad variety of goods (COSCA). After boiling or roasting, the new sliced tubers are consumed as a vegetable. Since the tubers degenerate quickly, they are cooked and beaten into paste, and are frequently used in soups and stews (“fufu” in Nigeria). Once they are harvested (postharvest physiological deterioration, PPD) occur. They are preserved as chip using solar energy and eaten after cooking or being ground into flour. Apart from processing cassava into foods, the crop can also be made into chips for animal feed and into starch for many food and non-food uses. Cassava flour is utilized in the production of glues, and pizza, biscuits, confectionery, pasta, and couscous-like goods. Cassava starch is used in the dairy, clothing, and paper manufacturing and in the making of plywood and veneer glues, and glucose and dextrin syrups. Through fermentation, cassava can be used for alcohol production and as a waste material can be processed to biogas (Kenyon et al., 2006). Several procedures comprising fermentation are useful for cassava postharvest preservation. Today, refining has been the most rampant technique of valuing cassava by-products. It enhances food security nutritious value, hygienic and sterile properties, energy density and organoleptic characteristics of diets (Guira et al., 2016) and facilitates transport and most importantly, detoxifies cassava roots by removing cyanogens (Nyirenda et al., 2011).

 

Cassava is traditionally processed into a great diversity of fermented products such as gari, lafun, attieke and chips which is suitable for transportation, trade and rapid preparation of meals (Koume, 2012). Fermented foods have become a significant aspect of the global diet as demand for their cultivation and use has risen dramatically over time (Ngobisa et al., 2015; Elyas et al., 2015). The fermentation method has been shown to be a suitable method to improve the safety, organoleptic and nutritional quality of many cassava derived foods (Oyewole, 1997). Increase variety in the diet improves active features and reduce anti-nutritional compounds (Ayoade and Sanni, 1992).

 

Although fermentation constitutes a significant process during the production, this still remain spontaneous and there is a dearth of evidence about the actual microorganisms that can be utilized as starter cultures. Optimization of fermentation process during the manufacture of indigenous food has been established and such practice will enhance the industrial take up of the native fermented foods with a view to support the nutritional intake of the people (Sanni, 1993; Holzapfel, 2002). Lactic acid microbes (LAB) are generally regarded as safe (GRAS), (Giraffa et al., 2010; Elyas et al., 2015). They play an important function in the majority of food fermentations and preservations and an extensive diversity of strains are routinely used as starter cultures in the manufacture of bakery products, dairy, meat and vegetable (Elyas et al., 2015; Gemechu, 2015). And further used as starters in fermented dough, alcohol beverages, probiotic animal feeds lactic acid fermentation of sorghum and maize-based cereals used as infant weaning foods (Wakil and Onilude, 2009). They assist in the improvement of cultured foods' descriptive and defensive attributes (Holzapfel and Wood, 2014). Lactobacillus plantarum has been used in food storage to extend stock life, add flavor, and achieve the perfect fragrance (Daeshel, 2004). During fermentation, L. plantarum has been linked to a reduction in anti-nutrients and waste substance in food (Smid et al., 2005). They have antimicrobial properties as a result of synthesis of compounds such as organic acids, carbon dioxide, hydrogen peroxide, diacetyl, and bacteriocins, which may prevent diseases and decomposition microbes, increasing the shelf life and improving the protection of fermented foods (Piard and Desmaxeand, 1992). The lactobacilli represent one of the major microbial groups included in the desirable fermentation.

 

The lactobacillus plantarum is regularly noted among the LAB during cassava dough fermentative germ (Edward et al., 2012) and lactobacillus platarum and lactobacillus fermetum have been recognized in the solid state fermentation of cassava through traditional gari production (Oguntoyinbo, 2007). Buyers are becoming more mindful of the practical and nutritious aspects of organic items, which has resulted in a rise in the commonness of healthy eating and the production of beneficial foods that fulfill basic nutritional needs (Nivien et al., 2016).

 

Consequently, there is a greater need to extract new LAB strains that can manufacture beneficial cannabinoid substances and have other distinct probiotic features. The 16s ribosomal RNA (rRNA) gene is one of the most common and faster approaches for verifying microbial (LAB) recognition in food. The prominence of the strategies for confirming the existence of LAB are focused on molecular biology. This method is focused on the sum of sequence similarities between various organisms, which shows how their genomes vary. Currently, more than 40% of cassava is refined into conventional food items.

 

There are opportunities to extend the traditional uses of cassava and introduce it into a widespread assortment of new food products, specifically in the urbanizing societies of the developing countries. Processing will also enable us to promote the use of composite flours from local crops in many food applications especially in manufacture of convenient foods to promote utilization of the white and yellow cassava varieties. Cassava flour can be supplement for production of baby food, glucose syrup and pastas as stated by (Nwabueze and Anoruoh, 2011).

 

Studies have also shown that cassava composite flour is a better supplement to wheat flour when paralleled to other root and tuber crops as reported by Olaoye et al. (2011). Hence, it is used in confectionaries making in the food industries. The use of cassava flour in food rations has clear advantages. Incorporating cassava into synthetic flour for fast food processing will lower costs and improve the quality of pasta, breakfast cereals, and pastries, among other products (Falade and Akingbala, 2009). Individuals, in addition to industries, bakers, and caterers, cultivate and buy cassava flour for use at home in the cooking of chin-chin, pie (meat and fish), buns, and cake, among other dishes.

 

1.1       STATEMENT OF PROBLEM

Cassava (Manihot esculeuta) is highly perishable after harvest and post-harvest losses are often substantial due to high moisture content. The bulkiness and high perishability of the harvested stored white and yellow cassava roots is major barrier to the wider utilization of the crop. Toxicity due to Cyanogenic glucosides is also a problem (Amoa-awua et al.,1996). Cassava contains toxic and anti-nutritional substances that interfere with digestion and up take of nutrients (Wobeto et al., 2007). Cassava is high in cyanogenic glucosides, which are harmful to humans and can lead to severe health problems.

To boost interest and raise revenue, it's essential to diversify the roots. Cassava roots are high in starch but low in protein and a number of important micronutrients. In Nigeria, the recently introduced yellow root cassava or β-carotene cassava cultivars are suitable for addressing vitamin A deficiency. More efforts are required to enhance the crop's use, particularly in the preparation of stiff dough (instant fufu) and snacks.

 

Lactic acid bacteria, yeast, and other bacteria lead significantly to starch oxidation, acidification, detoxification, and flavor production during cassava root fermentation (Oyewole, 1991). Lactic acid bacteria are useful in inhibiting spoilage of food and growth of pathogens, enhance the sensory value of fermented foods (Holzapfel and Wood, 2014), preventing diseases and promoting health. This study will reveal the role of the lactic acid bacteria in the modification of the white and yellow cassava flours.

 

1.2       JUSTIFICATION

Cassava (Manihot esculenta Crantz) is propagated in the tropical regions for its starchy roots. The roots are used for human intake, animal feed and as raw material in many industries. Processing of the roots helps to reduce post-harvest shortage and stabilizes seasonal fluctuations in the supply of the crop. Technologies of processing should be developed to enable the production of shelf-stable products which also will reduce post-harvest shortages and reduces the bulk to be transported and marketed, thereby, reducing transportation cost, and adding value at the rural areas. Nevertheless, it is important to establish strong relationship between small-scale cassava producers and new cassava products which is vital in creating awareness in cassava utilization (Dufour et al., 2002).

Fermentation is an essential processing method for the crop. Moreover, the fermentation method has been revealed to be a suitable method to improve the safety, organoleptic and nutritional quality of many cassava- based products. There is need to enhance the nutritional value of the white and β-carotene cassava (yellow cassava) fufu flour. The existence of pro-vitamin A (β-carotene) in the new cassava would improve the nutritional status of the consumers. Lactic acid bacteria have a standing history of application because of their beneficial effects on nutritional, shelf-life and organoleptic features of food.

 

There is need to evaluate the significance of the lactic acid bacteria in the modification of the white and yellow cassava flour. Constituents of cassava flour is essential in food industry as a result of its special characteristic which include clarity of appearance and low flavor. There seem to be limited literature on the fermentation of yellow cassava and white variety included in this research by diverse types of lactic acid bacteria (starter cultures) and their effects on nutritional, functional and sensory characteristics of the white cassava and newly bred yellow cassava flour, to obtain consistent product quality. Various food forms from these newly bred crops for value addition will enhance a wide range utilization of the crop and increase sales.

 

1.3       OBJECTIVES OF THE STUDY

The general objective of the study is to examine the influence of fermentation using diverse types of lactic acid bacteria on nutritional, functional and sensory characteristics of the white and yellow cassava fufu flour and to identify proposed starters from the natural fermentation, using 16s DNA sequencing.

 

1.3.1    Specific objectives

The specific objectives of the study include to:

        i.            Subject two varieties of cassava to natural and starter culture fermentation for                  72 h.     

      ii.            Determine the physico-chemical properties of the fermenting mash at different fermentation     periods (0h, 24h, 48h and 72h).

    iii.            Isolate and identify lactic acid bacteria (LAB) involved during spontaneous (traditional/ natural) fermentation of the cassava varieties using 16s DNA sequencing

    iv.            Produce cassava flours from the fermented cassava varieties

      v.            determine microbiological analysis of the fermented cassava flours

    vi.            determine the chemical composition and flavor profile of the fermented and non-fermented flours

  vii.            evaluate the sensory properties of the reconstituted cassava (fufu) flours.




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