MICROBIOLOGICAL AND NUTRITIONAL STATUS OF PALM WINE

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ABSTRACT


The microbiological and nutritional status of palm wine from Umudike environments was evaluated. About 20 samples of palmwine were used in this study. The total viable count of the bacteria from plamwine shows higher count at PLM10 (6.0x103cfu/ml) while lowest count was observed at sample PLM8 (2.4x104cfu/ml), higher counts were observed in PLM5while the yeast count has higher count at PLM 20 (2.6x103cfu/ml) while lowest count was recorded PLM3 (0.90x104cfu/ml). Five (5) microorganisms were isolated and they include; Staphylococcus aureus, Lactobacillus spp., Bacillus cereusStreptococcus spp and Escherichia coli.. Two (2) yeasts were isolated for this study and they include; Saccharomyces cerevisae and Candida sp  The temperature and pH of the palmwine the highest temperature (25oC) and pH (6.41). Other samples studied had other degrees of temperature ranging from 21oC -22oC. The proximate composition of the palmwine isolates showed that the sample are low in carbohydrate (1.01±0.01 - 2.12±0.02), crude protein (0.51±0.03 - 0.90±0.01), lipid (0.01±0.00 - 0.07±0.04), ash (0.31±0.11 - 0.78±0.06) and crude fibre contents (0.21±0.04 - 0.37±0.01). The moisture of the palwine was higher for all the samples ranging from 96.35±0.01 - 98.74±0.01. The vitamins in the palmwine were higher ranging from 21.48±0.02 - 22.76±0.08 for vitamin A and 6.60±0.80 - 7.42±0.02 for vitamin B. The mineral composition of the palmwine ranges from (3.24±0.28 - 3.85±0.02) for calcium; (30.02±0.00 - 31.35±0.01) for magnesium, (3.10±0.33 - 3.85±0.02) for iron; (0.25±0.04 - 0.31±0.03) for zinc and (0.28±0.08 - 0.48±0.08) for copper. The antimicrobial. The antimicrobial density test, Staphylococcus aureus was inhibited by all the antibiotics used in this study while E. coli, B. cereusStreptococcus spp and Lactobacillus spp showed other degrees of resistance and susceptibility.  Palm wine and its distillate are important solvent in herbal medicinal administrationTherefore, there is need to promote the quality of these products to enhance their nutritional and health benefits as widely consumed in this part of country.





TABLE OF CONTENTS

 

Certification                                                                                                                            i

Title page                                                                                                                                ii

Dedication                                                                                                                              iii

Acknowledgements                                                                                                                iv

Table of Contents                                                                                                                   v

List of Tables                                                                                                                          vii

Abstract                                                                                                                                  viii

CHAPTER ONE

1.0       Introduction                                                                                                                1

1.1              Specific objectives                                                                                                      4

CHAPTER TWO

2.0       Literature review                                                                                                         5

2.1       Biochemical constituents of palm wine                                                                      6

2.2       Palm sap/wine                                                                                                             7

2.3       Nutrient composition                                                                                                  9

2.4       Compositions in palmwine                                                                                          9

2.4.1    Sugars identified and their concentrations in palm wine                                            9

2.4.2    pH and organic acids concentration in palm wine                                                      10

2.4.3    Ethanol concentrations in palm wine                                                                          11

2.4.4    Minerals and trace elements present in the palm wine                                                13

2.4.5    Odorants of palm wine                                                                                               13

2.5       Microbial communities in palm wine                                                                          14

2.6       Methods employed for identification of microorganisms in palm wine                    16

2.7       Health implication                                                                                                       20

2.7.1    Electrolytes                                                                                                                 21

2.7.2    Magnesium                                                                                                                  21

2.7.3    Iron                                                                                                                             22

2.7.4    Manganese                                                                                                                  22

2.7.5    Water soluble vitamins, sucrose and proteins                                                             23

2.7.6    Thiamine                                                                                                                     23

2.7.7    Riboflavin                                                                                                                   24

2.7.8    Activation energies of thamine and riboflavin                                                           24

2.7.9    Ascorbic acid                                                                                                              24

2.7.10  Sucrose                                                                                                                        24

2.7.11  Protein                                                                                                                         25

CHAPTER THREE

3.0       Materials and methods                                                                                                26

3.1       Sample collection                                                                                                        26

3.2       Media to be used                                                                                                        26

3.3        Microbiological analysis                                                                                              26

3.3.1        Isolation of bacteria and yeast                                                                                                26

3.3.2        Identification of isolates                                                                                             27       

3.4       Staining techniques                                                                                                     27

3.4.1    Gram staining                                                                                                              27


3.4.2      Yeast viability staining                                                                                              28

3.4.3    Sugar fermentation                                                                                                     28

3.4.4    spore staining                                                                                                              28

3.5              biochemical test                                                                                                          29

3.5.1    Catalase                                                                                                                       29

3.5.2    Coagulase test                                                                                                             29

3.5.3    Citrate test                                                                                                                  29

3.5.4    Oxidase test                                                                                                                29

3.5.5    Indole test                                                                                                                   30

3.6       Determination of nutritional factors                                                                           30

3.6.1    Proximate analyses                                                                                                      30

3.6.1.1 Determination of moisture content                                                                             30

3.6.1.2 Determination of crude protein                                                                                  30

3.6.1.3. Determination of crude fat content                                                                           31

3.5.1.4 Determination of ash content                                                                                     32

3.6.1.5 Determination of crude fiber                                                                                      32

3.6.1.6 Determination of protein                                                                                           33

3.6.2    Determination of mineral elements                                                                             34

3.7       Antibiotic sensitivity testing                                                                                       34

CHAPTER FOUR

4.0       Results                                                                                                                        36

CHAPTER FIVE

5.0       Discussions, conclusion and recommendations                                                          45

5.1       Discussions                                                                                                                 48

5.2       Conclusion                                                                                                                  48

5.3       Recommendation

            References

 

 

 

 

 

 

 

 

LIST OF TABLES

Table                                                            Title                                                                  Page

4.1 shows the Morphological and biochemical identification of bacteria isolates from palm wine sample                                                                                                                                          34

4.2 shows the Morphological characteristics of yeast isolates from palmwine samples 35

4.3 shows the pH and temp. of palm wine samples after collection                                          36

4.4 shows the Microbial counts of palm wine samples (cfu/ml)                                                37

4.5 shows the Proximate composition of palwine                                                                     38

4.6 shows the Vitamin composition of palmwine                                                                      39

4.7 shows the Mineral composition of palmwine                                                                      40

4.8 shows the Antimicrobial susceptibility test                                                                         41

 

 




 

CHAPTER ONE


1.0       INTRODUCTION

Palm wine is made from the fermented sap of tropical plants of the palmae family, such as the oil palm (Elaeis guineensis), coconut palm (Cocus nucifera), date palm (Phoenix dactylifera), nipa palm (Nypa fruticans), kithul palm (Caryota urens), ron palm (Borassus aethiopum) and raffia palm (Raphia hookeri) (Dayo-Owoyemi et al., 2008). The unfermented sap is clear, sweet, colourless syrup containing about 10% - 12% (w/v) sugar, which is mainly sucrose (Ogbulie et al., 2007). Upon fermentation by the natural microflora, the level of this sugar decreases as it is converted to alcohol and other products whereas the sap becomes milky white due to the increased microbial suspension resulting from the prolific growth of fermenting organisms (Obire, 2005). It becomes a whitish, effervescent, alcoholic beverage consumed in very large quantities in West Africa, and known throughout the major parts of Africa under various names, such as “mimbo” in Cameroon, “nsafufuo” in Ghana, an “emu” in Nigeria and “bandji” in Côte d’Ivoire (Karamoko et al., 2012). It is widely consumed in tropical regions where palms grow such as Asia, South America, Africa and more particularly in Côte d’Ivoire amongst all socioeconomic groups. In such areas, the beverage plays an important role in the culture of the people and most African countries have their own palm wine, including Côte d’Ivoire (Ukhun et al., 2005). It has been associated to life farmer, because being less expensive and produced in the farming  surroundings. Indeed the consumed volumes are in the same way order of size than those of modern beers. Because of the nutritional wealth of this non fermented sap, it is often proposed to infants whose mother cannot produce the necessary milk for the nursing (Ezeagu and Fafunso 2003).

Generally, both brands of palm wine have several nutritional, medical, religious and social uses which have been reported elsewhere (Iheonu, 2000), to have increasingly enhanced the demand for this natural product. Although attempts has been made towards the preservation and shelf-life extension of palm wine through bottling, use of chemical additives and addition of plant extracts have greatly affected the organoleptic quality of the product (Obire, 2005). Several factors however have been adduced for this variation and they include the indigenous microbial flora, the biochemical composition of the two brands of palm sap, the tapping and post tapping processes.

In the study, microbiological quality of different palm wine conducted by Nwachukwu et al., (2016) revealed that the palmwine drink harboured several species of microbial genera which include Staphylococcus, Lactobacillus, Micrococcus, Serratia, Bacillus, Streptococcus, Saccharomyces cerevisae, and Candida tropiclis.

The palm sap of the palm tree is a rich medium capable of supporting the growth of several types of microorganisms like high numbers of aerobic mesophilic bacteria, coliforms bacteria, lactic acid bacteria, acetic acid bacteria and yeasts (Karamoko et al., 2012). Yeast populations have been reported in the palm wine in concentrations of about 104 to 107 cfu/mL, while, LAB ranging between 107 and 109 CFU/mL, AAB from concentrations of 105 to about 108 cfu/mL, total aerobic mesophiles ranging between 106 and 109 cfu/mL, and total coliforms have been reported in a range of 103 to 107 cfu/mL (Ouoba et al., 2012; Santiago-Urbina et al., 2013).

            Palmwine has several nutritional, medical, religious and socioeconomic uses which have increasingly enhanced the popularity and demand for this natural product (Ogbulie et al., 2007). The wine is rich in such nutrients as sugars, proteins, amino acids, vitamins and minerals (Ezeagu and Fafunso, 2003). Its residue (dregs) is rich in a dense population of yeasts (Bohoua, 2008) which are claimed medically to improve eye sight. The probiotic content of palmwine also bears on its nutritional value (Ezereonye, 2004). Palmwine contains about 10-12% sugar, mainly sucrose; about 0.36% protein; 10-19mg/100ml of vitamin C as well as about 160μg/ml of vitamin B12 (Okechukwu et al., 1984). Saccharomyces cerevisiae is the dominant yeast species responsible for the fermentation of palm wine tapped from Elaeis guineensis in Ghana and Cameroon, as well as Bandji in Burkina Faso (Ouoba et al., 2012). On the other hand, Saccharomyces chevalieri has been reported the yeast specie predominant in the Toddy from Sri Lanka. S. cerevisiae predominance in the palm wine production is attributed by the selective medium regarding pH, ethanol content, and anaerobic conditions, which favors the fermenting yeasts (Stringini et al., 2009). The major total volatiles and alcohols are produced by S. cerevisiae and S. chevalieri (Uzochukwu et al., 1999). e.g. the higher alcohols in fermented nipa (Nypa fruticans) sap is by cause of the metabolism of S. cerevisiae through two metabolic pathways; amino acids such as isoleucine and leucine, and glycolysis (Nur Aimi et al., 2013). Moreover, Zymomonas mobilis is also considered as the microorganism responsible for the palm wine fermentation and has been reported in Taberna, considering that this microorganism has ability to grow in acidic condition (pH about to 3.53) and tolerate high ethanol concentration (10.33% v/v) (Alcántara-Hernández et al., 2010), similar results are reported in palm wine obtained by “inflorescence tapping” from Elaeis guineensis in Nigeria (Obire, 2005).

Other identified yeasts during the tapping process probably play a determinant role in the fermentation. e.g. in wine fermentation is reported that the apiculate yeasts such as Hanseniaspora guilliermondii and Hanseniaspora uvarum have the capacity to influence, in a positive way, the aromatic profile of wines. H. guilliermondii has been reported to produce high levels of 2-phenylethyl acetate and 1-propanol (Moreira et al., 2011).

Thus also, the predominant LAB reported in palm wine fermentation are Lactobacillus plantarum and Leuconostoc mesenteroides (Amoa-Awua et al., 2007). These microorganisms are responsible for the sour taste of palm wine and are responsible for the pH decrease during the tapping through the organic acids production (Amoa-Awua et al., 2007; Ouoba et al., 2012). These bacteria also control the growth of undesirable microorganism such as enterobacteria by acid and H2O2 production (Alcántara-Hernández et al., 2010) e.g. Santiago-Urbina et al., (2013) reported that the total coliforms population in Taberna decreased during the tapping period with the increased of lactic acid production. Similar results are reported in Bandji where the predominant genus is Lactobacillus representing 86.67% of the LAB total isolates followed by the genera Leuconostoc (10%).


1.2              SPECIFIC OBJECTIVES

i.                    To isolate, characterize and identify microorganism resident in palm-wine from umudike environments.

ii.                  To isolate, characterize and identify yeasts resident in palm-wine from umudike environments.

iii.                To determine the nutritional status of palm-wine from umudike environments.



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