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Abstract

Kunu or kunuzaki is a beverage drink made from grains such as millet, sorghum and maize or other
combinations. It is a nonalcoholic beverage widely consumed in Nigeria. It is marketed in public placed
such as offices, markets, schools, motor parks. This work is done on Bacteriological Examination of kunu
drinks sold in Aba metropolis in Aba State, Nigeria. Twenty kunu drink samples were purchased in
different markets in Aba namely Ahiankwo, Ahiaohuru, Ariaria, Tonimas and Osisioma makers.

Nutrient agar and pour plate methods were used bacterial isolation. Also, phenotypic and Biochemical
characterization methods were employed to further identified bacteria isolates. The bacteria isolated
from kunu were Staphylococcus aureus which had the highest percentage occurrence as 46.7%,
followed by Escherichia coli ( 25% ), Salmonella species (15% ) and Streptococcus faecalis (13.3% ).

The pH of the sampled kunu drinks ranged from pH 3.80 to pH 5.00. All the kunu drink samples
examined had varying levels of bacteria contamination ranging from 1.1×10^4cfu/ml to
18.5×10^4cfu/ml and total coliform count ranging from 2MPN/ml to 15MPN/ml which exceeded the
recommended safe level of zero coliform organisms detectable per 100ml. From our present findings, it
reveals that kunu drinks retailed and sold in these markets constitute main concern for public health as
these can cause a variety of infections of food intoxications. Thus, there is a need to establish a system
of monitoring the market where kunu is sold and post production preservation and packaging
techniques to make sure that it is safe for human consumption.

Table Of Content

Preliminary Page(s)

Tittle page

Declaration

Dedication

Acknowledgement

Table of content

List of tables

Abstract

Chapter One

1.0 Introduction
1.1 Justification

1.2 Significance

1.3 Aim of study

1.4 Specific Objective

1.5 Scope of the study

1.6 Nutritional value of kunu

Chapter Two

2.0 Literature Review

2.1 Historical background of kunu

2.2 Local production of kunu

2.3 Cereals used in kunu production

2.3.1 Millets

2.3.2 Sorghum

2.3.3 Maize

2.4 Ingredients used in kunu production

2.4.1 Ginger

2.4.2 Black pepper

2.4.3 Garlic

2.4.4 Sweet potatoes

2.4.5 Malted rice

2.5 Health benefits of kunu drink

2.6 Biochemical analysis

2.7 Bacteriological analysis

Chapter Three

3.0 Materials and Methods


3.1 Study design

3.2 Study area

3.3 Collection of samples

3.4 Microbiological examination

3.5 Phenotypic characterization

3.5.1 Cultural morphoy

3.5.2 Gram staining

3.6 Biochemical characterization

3.6.1 Motility test

3.6.2 Catalase test

3.6.3 Ciagulase test

3.6.4 Indole test

3.6.5 Oxidase test

3.6.6 Citrate utilization test

3.6.7 Methyl red test

3.6.8 Voges proskaeur test

3.6.9 Sugar fermentation test

3.7 Total bacteria count

3.8 Total coliform count

3.9 Statistical analysis

Chapter Four

4.0 Results
Chapter Five

5.0 Discussion

Conclusion

Reference

Appendix

List Of Tables

Table 4.1 Phenotypic and Biochemical Characterization of Bacterial isolates from kunu drinks.

Table 4.2 Percentage Occurrence of Bacterial isolates.

Table 4.3 PH values of kunu drinks.

Table 4.4 Total Bacterial count if kunu drinks.

Table 4.5 Total coliform count of kunu drinks.

Chapter One

1.0 Introduction

Kunu is an important nonalcoholic beverage mostly found in Northern Nigeria. It is prepared from grain
such as Sorghum, Millet, Maize or Wheat (Gaffa et al ., 2002). Apart from these cereals, kunu has shown
to be produced from tigernuts (Belewu and Abodunrin, 2006), guinea corn and rice (Umaru et al., 2014).
Because kunu is prepared in traditional method, the ingredient concentrations are neither quantified
nor standardized (Aboh and Oladosu, 2014). The production procedures varies depending on household,
taste and cultural habits of the consumers. This leads to variation in the taste, quality and specification
of the product.

According to Umaru et al., 2004, the processes involved in the preparation of kunu zaki include: Steeping
the whole grains in water for 6-24 hours followed by wet milling usually withspices and sweet potato,
gelatinization of a portion of the mixture in hot water and then pitching with about one quarter fresh
(ungelled) part of the mixture. Spices such as ginger, alligator peppe, red pepper, black pepper and
Kakandoru or Eru are usually utilized (Adelekan et al., 2013). After preparation, it can be served at home
and public places such as offices, markets, schools, motor packs and a very common consumed beverage
in occasions such as weddings, naming ceremonies, birthday celebrations, burial etc.(Pasqualon et al.,
2018), where it is served as food appetizer, refreshing drink and complementary food for infants
(Olasupa et al., 2002).

Nutritionally, kunu is made up of 11.6% protein, 3.3% fat, 1.9% ash, 82-92% moisture, 76.8%
carbohydrates and arrays of amino acid ( Lichtenwalner et al., 1979). Apart from the nutritional benefits
of kunu consumption, the drink has been shown to have other benefits including reduction in blood
cholesterol, lowering the risk of diabetes and prevention of blood clot formation (Ofudje et al., 2016).
Because of the traditional method employed in the preparation of kunu, the
unregulation/unstandardization of ingredients used and possible low sanitary procedures employed in
the preparation, kunu may provide an ideal environment for the growth of microorganisms and its
consumption leading to food borne intoxication/poisoning. Various studies have investigated the
microbial composition of kunu zaki in various parts of Nigeria with varying results.

Osuntogun and Aboabo, 2016, reported that kunu contains lactic acid bacteria (LAB) including
Lactobacillus species, Streptococcus species and Leuconastic species which can cause food borne
diseases. Also, coliforms, molds and yeast causes spoilage in the drink thereby producing undesirable
changes (Uche et al., 2012). Food pathogens such as Escherichia coli have been implicated in food
poisoning resulting from their consumption (Famine et al., 1989). Kunu zaki contamination with
pathogenic bacteria is of public health importance and might cause diverse food related illness and
infection to consumers and therefore, proper hygienic preparation is to be ensured ( Elijah et al., 2017).
The effect of storage on the drink varies and the time lag during which the drinks lose their nutritional
properties vary, hence there is a need to know the more appropriate method to storage, whether
ambient or refrigerated to reduce the incidence of certain diseases (Tamine et al., 1989).

1.1 Justification Of The Study

In developing natiobslije Nigeria, it has not been possible to have control over the processing of hawked
foods because most of the vendors lack the adequate knowledge of food processing and adequate
handling practices. As such, there is likely to be a high risk of chemical and microbial contamination. A
large number of lactic acid bacteria, coliforms, molds and yeast have been reportedly implicated in food
spoilage as the used the carbohydrate content of foods for undesirable fermentation processes ( Odunfa
1988; Ojoko et al., 2002; Amusa et al., 2005). Therefore, it becomes very necessary to conduct this
research to determine the bacteria load, nutritional and microbiological qualities of these non-alcoholic
drink called kunu zaki in Aba, Abia State.

1.2 Significance Of Study

The significance of this work is to enable those producers to improve the hygienic conduction handling
of kunu zaki and a good knowledge of safe food handling practice, suitable protective clothing to be
worn and it will also enable the public to be aware of various pathogenic organisms present in kunu
hence increasing the health awareness on the dangers of drinking or taking kunu zaki. As a result of this,
the prevention of contamination should be adhered to.

1.3 Aim Of The Study

This work is aimed at Bacteriological examination of kunu sold in Aba.

1.4 Specific Objectives

To isolate and characterize microbial species associated with kunu.

To determine total bacteria count of kunu.

To determine total coliform count of kunu.

To determine the pH of kunu.

1.5 Scope Of The Study

This research work is limited to the comparative analysis of Bacteria load of samples of kunu (gruel)
found in Aba major markets.

1.6 Nutritional Value Of Kunu

Kunu zaki processed from sorghum grains contains 11.6% protein, 3.3% fat, 1.9% ash and 76.8%
carbohydrate and arrays of amino acid (Lichtenwalner et al., 1979). However, this non-alcoholic
beverage is however becoming more widely accepted in several other parts of Nigeria, including Aba
metropolis, port Harcourt metropolis etc, owning to its refreshing qualities.

The preparation of this beverage has become technology in many homes in the rural communities and
more recently in the Urban areas where commercial production due to support from the government
through the poverty alleviation scheme, has helped to alleviate poverty among the people
(Lichtenwalner et al., 1979).

Chapter Five

5.0 Discussion

5.1 Bacterial Isolates From Kunu Drink Samples


The result of this study showed that the bacterial load of the sampled kunu drinks includes Escherichia
coli, Staphylococcus aureus, Streptococcus faecalis and Salmonella species ( table 4.1 ) with
Staphylococcus aureus having the highest percentage occurrence of 46.7% followed by Escherichia coli
(25%), then by Salmonella species (15%) and the least was Streptococcus faecalis (13.3%) ( table 4.2).
This result agrees with the findings of Eden et al., (2017) who isolated Salmonella species, Escherichia
coli, Staphylococcus aureus, Streptococcus species amongst others from kunu zaki sold in Eiyenkorin,
Kwara state, Nigeria. In other parts of Nigeria, similar bacteria were isolated from kunu by other
researchers such as Salmonella species, Staphylococcus aureus, Streptococcus species and Bacillus
species ( Etang et al., 2015 ), Escherichia coli, Staphylococcus aureus, Salmonella species and Shigella
species in Maiduguri ( Ogbonna et al., 2011), while some reported Escherichia coli, Staphylococcus
aureus, Streptococcus pyogenes ( Edward and Ohaegbu, 2012). Most researchers had comparable
bacteria in Port Harcourt ( Essien et al., 2009 ), Oyo and Lagos ( Amusa et al., 2009 ).

The organisms isolated from kunu zaki retailed in Aba major markets are of public health concerns these
bacteria genera are known to cause a variety of human illnesses and have been implicated in cases of
food intoxication. The presence of Escherichia coli, Streptococcus faecalis and Salmonella species
indicates possible feacal contamination and unhygienic preparation process of the drink. This is because
part of the preparation process of kunu is cooking which should eliminate most of the pathogens
isolated in this work, thus indicating that the contamination recorded was after the cooking process
when the drink most have cooled down Anumudu and Anumudu, 2019).

Among the bacteria isolated, Staphylococcus aureus was the most prevalent. Staphylococcus aureus is
normal inhabitant of human body which can be found on the skin, mouth, nostril, hands, various
surfaces etc. These were possible sources from where Staphylococcus aureus can contaminate kunu
during processing and packaging. Bacteria might be present in storage containers, sieve used to filter the
finished product and contamination from handlers ( Ikpo et al., 2013).

5.2 pH Values Of Kunu Drinks

The pH of samples ranged between pH 3.80 to pH 5.00 including acidity. The result agrees with the pH
range of 3.80 to 3.99 reported by Innocent et al (2011), and pH 3.83 to 5.65 reported by Amusa and
Ashaye (2009). The acidity of the kunu drink may be due to the presence of some bacteria which help in
fermentation of kunu products ( Ekanem et al., 2018).

5.3 Total Bacterial Count Of Kunu Drink


All kunu drinks sampled were contaminated with varying levels of bacterial count ranging from
1.1×10^4cfu/ml to 18.5×10^4cfu/ml in which samples from Ahiankwo had the highest total bacterial
count ( 4.5×10^4 — 18.5×10^4cfu/ml). Our findings have some similarity with various studies. For
instance, Anumudu and Anumudu (2019), reported a total bacterial count ranging from 1.4×10^3cfu/ml
to 4.4×10^4cfu/ml in kunu drinks sold in Owerri metropolis, Nigeria. Sule et al (2016) reported a total
bacterial count ranging from 1.5×104cfu/ml to 7.4×10^4cfu/ml. Slight variation that exist in the findings
of this study when compared with previous studies could be due to handling period, quality of the
materials used for production and hygienic status of the processors and vendors. Our study reveals a
high level of contamination, this is very dangerous o the health of the public as most strains of the
isolates are toxic and their detection in kunu drinks posses a serious health effects to the consumers.
Also, it was noted that local drinks such as kunu may act as vehicle for the transmission of zoonotic and
bacterial infections such as Staphylococal infection, Salmonellosis, Shigellosis, Tuberculosis, Listeriosis
etc ( Umaru et al., 2014). In the other hand, preparation of kunu in environment with poor sanitation
conditions predisposes the preparation and packaging processes to contamination and exposes the
consumers to health risk.

5.4 Total Coliform Count

The occurrence of coliform in kunu drinks is as a result of faecal contamination of the drinks. All kunu
drinks examined had coliform count ranging from 2MPN/ml to 15MPN/ml. The sample with the highest
most probable number is Sample 1 from Ariaria market with 15MPN/ml and the least is sample 4 from
Ahiankwo market with 2MPN/ml . Our result is in line with Umar et al., (2014) who had coliform count
ranged from 6.1— 8.2MPN/ml in their findings from kunu drinks sold at Umaru Musa Yar’adua
University Campus, Kastina. The coliform count is above the zero value recommended for safe water by
Saskatchewan drinking water quality Standard and health Canadia guild lines for Canadian drinking
water quality which stated that the maximum concentration (MAC) for coliform in drinking water is zero
organisms detectable per 100ml ( http/www.health.gov.SK.can/lab). This renders the sampled kunu
drinks bad for human consumption. The presence of coliform such as Escherichia coli may be as a result
of acid tolerance response mechanism they often develop. Also, the presence of coliform bacteria in
kunu will be of public health concern because teaming populace, especially students, relies on these
drinks as cheaper alternative to the bottled soft drinks ( Tamine et al., 1989). The bacteria isolated from
this work are known pathogens responsible for millions of cases of gastrointestinal diseases and death
each year.

Conclusion

The results of this study showed that the bacterial load of locally prepared kunu zaki drink in Aba major
markets is high due to possible use of contaminated water in its preparation and improper handling. The
most predominant bacterial contaminant is Staphylococcus aureus with a percentage occurrence of
46.7%, followed by Escherichia coli ( 25% ), then by Salmonella species and the least being Streptococcus
faecalis ( 13.3% ). As a result, these microorganisms can cause varying levels of diseases including
diarrhea, food intoxication amongst others. The total coliform count us above the zero level
recommended for safety which makes this drink dangerous and unsafe to the public. The occurrence of
this pathogens can be prevented by employing good manufacturing practice. From our research, the
production of kunu as non- alcoholic beverage for public consumption should be regulated by
appropriate regulatory agencies such as National Agency for Food and Drug Administration Control
( NAFDAC ) and other regulatory bodies, to reduce the risk of consumers infection. Producers should be
made to have fair knowledge of food preservation and food sanitation.

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