ABSTRACT
This study evaluated the microbiological quality of fruits sold in Umuahia metropolis. A total of fifteen (15) sliced fruit samples consisting of 5 watermelon, 5 pawpaw and 5 pineapples were collected from five different markets. The samples were cultured on Nutrient Agar Medium, MacConkey Agar, and Sabauroud Dextrose Agar using pour plate techniques. A total of nineteen (19) bacteria were isolated and identified using morphological characteristics, pigmentation on media, microscopy, biochemical and sugar fermentation methods. The bacteria were Bacillus species, Escherichia coli, Staphylococcus aurues, Proteus species and Micrococcus species while the fungal isolates were Aspergillus niger, Aspergillus fumigatus and Rhizopus stolonifer. Ndioru had the highest aerobic plate count at 3.1x105, followed by Unuariaga (3.0x105), Orieugba (2.9x105), Gate six (2.4x105), while the least aerobic plate count was recorded at Ahieke (1.9x105) in that order. The total coliform plate count investigated reveals that Orieugba had the highest coliform plate count at (3.5 x105) while Ndioru and Umuariaga had the least total coliform plate count of (2.4 x105). The total viable fungal plate count reveals Gate six as having the highest viable fungi count at (4.5 x105), while Ndioru has the least viable fungi count at 2.3x105. The percentage occurrence and distribution of bacterial isolates revealed that Escherichia coli occurred most frequently with percentage occurrence of 6(20.0%) followed by Bacillus spp and Proteus spp at 4(13.3%) each, Staphylococcus aureus 3(10.0%), and Micrococcus spp 2(6.6%) respectively. Aspergillus niger was the most predominant with 5(16.6%), followed by Aspergillus fumigatus 4(13.3%) while the least was Rhizopus stolonifer at 2(6.6%).This study also revealed that Staphylococcus aureus showed high level of susceptibility to Amoxil, Erythromycin and Levofloxacin. Bacillus spp showed high level of resistance to Ciprofloxacin, Amoxill and Chloramphenicol while Escherichia coli showed high resistance to Norfloxacin. The presence of these microorganisms in the fruit juices is of public health significance. Therefore, regular monitoring of the quality of commercially available fruit juices for human consumption is recommended to avoid outbreak of food borne illness resulting from the organisms encountered in this study.
TABLE OF CONTENTS
Title
Page i
Certification ii
Dedication iii
Acknowledgements iv
Table
of Contents v
List
of Tables viii
Abstract ix
CHAPTER ONE
1.0 Introduction
1
1.1 Aim and Objectives 3
CHAPTER TWO
2.0 Literature Review 4
2.1 Microbiology
Of Polyethylene-Packaged Sliced Watermelon
(Citrullus
Lanatus) Sold By Street Vendors in Nigeria 4
2.2 Microbiological
Quality of Pre-Cut Fruits On Sale in Retail
Outlets
in Nigeria 5
2.3 Microbiological
Quality of Sliced Fresh Fruits Sold In Bida Nigeria 6
2.4 Bacterial Pathogens Associated with Fruit
Contamination 7
2.4.1 Campylobacter 7
2.4.2 Pathogenic
Escherichia coli 8
2.4.3 Salmonella
spp 8
2.4.4 Shigella
spp 9
2.4.5 Staphylococcus 10
2.4.6 Vibrio 10
2.4.7 Listeria
monocytogenes 11
2.4.8 Sporeformers 11
2.5 Sources of Contamination 12
2.5.1 Production Environment 12
2.5.2 Postharvest Handling 13
2.5.3 Human Hygiene 14
2.6 Pathogen Prevention 15
2.6.1 Processing Strategies 15
2.6.2 Handling Parameters 17
2.6.3 Alternative Technologies 18
CHAPTER THREE
3.0 Materials and Methods 20
3.1 Study Area 20
3.2 Materials
and Reagents 20
3.3 Collection
of Samples 20
3.4 Sterilization of Materials 21
3.5 Preparation of Culture Media 21
3.6 Inoculation and Isolation 21
3.7 Purification
of Isolates 22
3.8 Identification of the Isolates 22
3.9 Gram
Staining 22
3.10 Biochemical Test 23
3.10.1 Catalase Test 23
3.10.2 Indole Test 23
3.10.3 Citrate Utilization Test 23
3.10.4 Hydrogen Sulphide (H2S) Production
Test 24
3.10.5 Starch Hydrolysis 24
3.10.6 Motility, Indole, Urease (MIU) 24
3.10.7 Coagulase Test 25
3.10.8 Oxidase Test 25
3.11 Identification of Fungal Isolates 25
3.11.1 Colonial Morphology 25
3.11.2 Yeast Viability Staining 26
CHAPTER FOUR
4.0 Results 27
CHAPTER FIVE
5.0 Discussion and Conclusion 34
5.1 Discussion 34
5.2 Conclusion
and Recommendation 37
LIST OF TABLES
|
TABLES
|
TITLE
|
PAGE NO
|
|
1
|
Total
Viable Bacterial Mean Counts from selected sliced fruit samples
|
27
|
|
2
|
Total
Viable Bacterial Mean Counts from selected sliced fruit samples
|
28
|
|
3
|
Cultural Morphology and
Microscopic Characteristics Fungal Isolates from
the selected sliced fruit samples.
|
29
|
|
4
|
Distribution
of Bacterial and Fungal Isolates from the Selected Pineapple Juice Samples
|
30
|
|
5
|
Antimicrobial Sensitivity Patterns of Bacterial
Isolates from Selected Pineapple Juice Samples
|
31
|
CHAPTER ONE
1.0 INTRODUCTION
Fruits
are an extraordinary dietary source of nutrients, micronutrients, vitamins and
fiber for humans and are thus vital for health and wellbeing. Well balanced
diets, rich in fruits and vegetables, are especially valuable for their ability
to prevent vitamin C and A deficiencies and are also reported to reduce the
risk of several diseases (Daniel et al.,
2014). Fruits product are known to carry natural nonpathogenic micro flora;
however, contamination with pathogens from humans or animals source can also
occurs sporadically at various stages along the production. Globalization of
fresh fruits potentially increase human exposure to a wider variety of
food-borne pathogens and has resulted in a potential increased risk of
food-borne illness outbreak associated with the consumption of these raw
commodities. These pathogens may invade the inner surface of the fruits during
slicing or peeling. Studies have already confirmed that pre-cut fruits might be
contaminated with Salmonella, Listeria monocytogens and high
count of Stapyylococcus aureus (Eni et al., 2010). Consumption of
sliced products has been on the increase locally and globally. This is so
because they are more convenient, easily accessible, and most especially
cheaper than the whole fruits (Nwanchukwu et al., 2008). The increase in
the consumption of sliced fruits has been linked with parallel increase in
food-borne illnesses (Estrada-Garcia et al., 2004; Eni, et al.,
2010). Sliced fruits commonly consumed in Borno state include pawpaw pineapple
and watermelon. Pawpaw pineapple and watermelon are process and sold by street
vendors with poor education levels and untrained in food hygiene (Barro et
al., 2006). The consumption of sliced pawpaw and watermelon may have
potentially increase the risk of food-born disease caused by variety of
pathogens, cross contamination of fruits, unsanitary processing and use of
dirty trays for display of fruits further increases the risk of contamination.
Another major source is the water used in washing those cut fruits (Estrada-Garcia
et al., 2004). Bacteria causing gastrointestinal infection can
contaminate the sliced fruits thus exposing the consumer to greater risk (Eni, et
al., 2010).
In
Nigeria, fruits are popularly displayed completely exposed for sales in
shopping malls, along busy and major streets and hawked by street food vendors
in motor parks and on busy roads with heavy traffic, security check points or
at bad spots on the highways where motorists are forced to slow down. Fruits
are often purchased as ready to eat and thus usually consumed without washing.
The sliced fruits such as pawpaw, pineapple and watermelon could become
contaminated with foodborne bacteria in the process such as processing utensils
in adequate hygienic conditions, as they are left uncovered, trays left open in
unsuitable places for buyers and irregular hand-washing by the vendors. The
consumption of sliced pawpaw pineapple and watermelon may have potentially
increased the risk of food-borne disease caused by variety of pathogens. To
investigate the quality of those consume fruits sold at road side is of great
concern (Eni, et al., 2010). The increased consumption, coupled with the
associated risk of disease to which consumers may be exposed, is a matter of
great concern. It is difficult for one to attest to the hygiene of the
processors or to the sanitary conditions at points of preparation (Barro et al., 2006). Moreover, the case is
worsened by the fact that sliced fruit street vending is done without adequate
storage conditions, thereby exposing the sliced fruits to flies and other
disease-causing agents. The sliced/peeled fruits are processed and sold by
unlicensed vendors with poor education levels and untrained in food hygiene (Barro
et al., 2007). The consumption of
sliced/peeled fruits may thus potentially increase the risk of food-borne
diseases caused by a wide variety of pathogens (Oranusi et al., 2004). There are different sources of microbial invasion
of sliced produce. Pathogens may invade the interior surfaces of the produce
during washing, peeling, slicing, trimming, packaging, handling and marketing.
The use of dirty utensils, as well as the open display of street food produce
encourages sporadic visits by flies, cockroaches, other insects, and dust (Chukwu
et al., 2010). Holding of sliced
fruits that requires no further processing before consumption at ambient
temperatures during retail maintains the produce at optimum temperatures for
proliferation/invasion by pathogenic mesophiles (Barro et al., 2006). Bacteria like Salmonella
spp., Shigella spp., Campylobacter spp. and Escherichia coli can
contaminate sliced fruits through contact with sewage and contaminated water (Oranusi et al., 2004). Fruits have been
associated with outbreaks of food-borne disease in many countries. Organisms
involved include bacteria, fungi, viruses and parasites (Chukwu et al., 2010).
1.1 AIM AND OBJECTIVES
The
aim of this study is to evaluate the microbiological quality of fruits sold
within Umuahia metropolis, while the specific objectives are;
·
Determination of total
aerobic bacteria plate count and total fungal count.
·
Isolation and
identification of bacteria and fungi contaminants from different vended fruits
within Umuahia.
·
Determination of the
percentage frequency distribution of the microbial isolates on the vended
fruits.
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