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 cereus, Streptococcus 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. cereus, Streptococcus spp and Lactobacillus spp showed other degrees of resistance and susceptibility. Palm wine and its distillate are important solvent in herbal medicinal administration. Therefore, 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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