DETERMINATION OF NICOTINE CONTENT AND MICROBIOLOGICAL EXAMINATION OF POWDERED TOBACCO SOLD IN UMUAHIA, ABIA STATE

ABSTRACT

Powdered tobacco (snuff) is a finely ground product of Nicotina tabacum leaves that is taken by inhalation through the nose or by tucking it between the lower lips and the gum. The principal alkaloid found in powdered tobacco is nicotine and the determination of the nicotine content was done using the alkaline precipitation gravimetric method and the result obtained from the snuff samples ranged from 0.3mg- 0.8mg. Identification of microorganisms present in the snuff sample sold in various markets in Umuahia. Abia state was carried outusing standard microbiological techniques. The organisms identified were Staphylococcus, Aspergillus, Bacillus, Streptococcus, Micrococcus, Rhizopus and Penicillium. The mean total bacterial counts obtained from various market in Umuahia are as follows, Ndoru ranged from 3.67x10⁴± 0.58 – 5.33x10⁴± 0.58cfu­/g,while Ubani ranged from 2.67x10⁴± 0.58 – 4.33x10⁴± 0.58 cfu­/g. Orie-ugba ranged from 4.00x10⁴± 1.00 – 5.33x10⁴± 1.53 cfu­/g and Ahiaeke ranged from 2.67x10⁴ ± 0.58 – 5.00x10⁴ ± 1.00. The fungal counts for Ndoru ranged from 0.67x10²± 0.58 – 2.00x10²± 1.00 cfu­/g while Ubani ranged from 1.00x10²± 0.00 – 2.67x10²± 1.15 cfu­/g and Orie-ugba ranged from 1.67x10²  ± 0.58 - 2.67x10² ± 0.58 cfu­/g and Ahiaeke ranged from 1.67x10² ± 0.58 - 2.33x10² ± 0.58 cfu­/g. Staphylococcus had the highest  occurrence among the bacterial isolates (33.3%), while Streptococcus had the least (16.7%). Aspergillus had the highest occurrence among the fungi (40%), whileRhizopus had the least (25%).Most snuff samples sold in markets in Umuahia were contaminated by potentiallypathogenic microbes. Good manufacturing practices and improvements in the sanitary condition

TABLE OF CONTENTS

Title Page                                                                                                i

Certification                                                                                             ii

Dedication                                                                                              iii

Acknowledgements                                                                      iv

Table of Contents                                                                                v

List of Tables                                                                                       viii

Abstract                                                                                                  ix

CHAPTER ONE                                                                                            

1.1       Introduction             1                                                                        

1.2        Aim                                                            2                                 

1.3       Objectives                                           2                                                                                                                                                  

CHAPTER TWO

LITERATURE REVIEW                                                                  

2.1       Tobacco                                                                                                                     3

2.2       Classification of Tobacco Plant                                                                              12

2.2.1    Family Salanaceae                                                                                                    12

2.2.2    Nicatiana Rustica                                                                                                              12

23        Chemical Composition of Tobacco                                                                                    13       

2.4       Short Term Effects of Use of Powdered Tobacco (Snuff)                                          14

2.5       Long-term effects of smoking or chewing tobacco                                                    14

2.5.1     Heart problems                                                                                                                                                      14       

2.5.2     Cancers                                                                                                                             14       

2.5.3     Lung disease                                                                                                                                             15      

2.5.4     Other health problems                                                                                                      15

2.6         Effects of tobacco on the brain                                                                                                   15

CHAPTER THREE   

MATERIALS AND METHODS

3.1     Source of materials                    17       

3.2    Determination of alkaloid content (nicotine)                                                             17

3.3     Sample preparation                                                                  18

3.4     Media preparation                                                                                                               18

3.5     Determination of microbial load                                                                               19 

3.6     Isolation and identification of microbes                                                                   19                                                                                                                                                                                           

3.7     Characteristics of isolates                                                                                                   20

3.7.1   Characterization of fungi isolates                                                                                       20

3.7.1.1    Colony features                                                                                                              20

3.7.1. 2   Structural features                                                                                                         20

3.7.2     Characterization of bacteria isolates                                                                      21

3.7.2.1   Colony features                                                                                                                21

3.7.3     Biochemical tests                                                                                                               21

3.8       Identification of isolate                                                                                                        24

CHAPTER FOUR

4.1   Results                                                                                                                        25                                                                                                             

CHAPTER FIVE

5.1     Discussion                                                                                                                            31

5.2     Conclusion                                                                                                                           32

5.3      Recommendation                                                                                                                32

 REFERENCES

                                                        LIST OF TABLES                   

TABLE              TITLE                                    PAGE          

 1                   SHOWS THE CONCENTRATION OF NICOTINE IN                            26                  

                       EACH OF THE SNUFF SAMPLES

2                  BIOCHEMICAL CHARACTERISTICS OF EACH OF                                      27

                       THE ISOLATES          

 3                 MEAN TOTAL MICROBIAL COUNTS FOR DIFFERENT

                    MARKETS (CFU/G)                                                                                                                                                      

4             PERCENTAGE OCCURRENCE OF EACH OF THE BACTERIAL          29

                    ISOLATES

 5           PERCENTAGE OCCURRENCE OF EACH OF THE FUNGAL               30

                   ISOLATES

CHAPTER ONE

INTRODUCTION

Powdered tobacco (snuff) is a finely ground product of Nicotina tabacum   leaves that is taken by inhalation through the nose or by tucking it between the lower lips and the gum. (Ayo-yusuf et al., 2005).It has being recommended as nicotine substitute of cigarette, it is

locally called   Anwuru by the Igbos,   Saara by the Yorubas and Kirha or Taban hanchi by the Hausas. Snuffing has become quite popular as a medication for long grief, pain and aches (Tuner et al., 2011).Snuff is also available in wet form. Wet snuff sometimes called snus is rubbed inside the mouth instead. In other words, "Snuff" is tobacco that has been dried and processed into a powder. This powder is snorted into the nose, where it is absorbed through the nasal passage. There are various processes involved in the manufacturing of tobacco powder (snuff) however, in Nigeria the powder is produced by grinding the cured tobacco leaves with locally made mortar or by mechanical grinding (Ogundaro1980) Also depending on the scale of operation, grinding stones may be used. It can also be mixed with potash depending on the people's culture. This enhances flavour quality of the tobacco powder. In the United States of America tobacco powder is produced from dark cured and fire cure types of leaf. It involves steaming of the leaves, chopping the leaves and packaging in hogshead or other container for fermentation to take place. This process requires about two months for completion. After fermentation, the tobacco looses the creaosote- like odour and becomes more aromatic the product is then dried by passing through heated container and finally reduced to powdered form by using steel drums fitted with rotating rollers. People take snuff for different reasons, for medicinal purposes, for smoking cessation among others etc (Ureme et al.,2007).The principal content of tobacco is nicotine others include nitrosamines, nitrosamine acids, polycyclic aromatic hydrocarbons , aldehydes and tar(Fant et al., 2009) Many of these chemicals are known to cause cancer and various other ailments. In Igbo communities of Nigeria where tobacco is utilized for cultural and traditional purposes, Nicotine has been associated with addiction in regular smokers and snuffers (Ureme et al., 2007). There are many microorganisms that are associated with snuff mostly transferred during production due to poor sanitary conditions prevailing around the snuff mills and market. This can expose snuff to high levels of microbial contamination. Some of the health effect of snuff use include increase heart beat and raised blood pressure, development of oropharyngeal and upper respiratory tract cancer (gingival recession carries staining and abression), cardiovascular diseases and adverse reproductive outcome (Russell et al., 2010). Snuff is taken raw without further processing to reduce the microbial load mostly by elderly people with lower immunity and other health conditions

1.1       Aim    

This study is aimed at determining the Nicotine content and also determining the presence of microorganisms (bacteria and fungi) in powdered tobacco sold in Umuahia  Abia   State.

1.2       Objectives

·       To measure the nicotine content of the powdered tobacco

·       To isolate and identify the different bacteria present in the powdered tobacco

·       To isolate and identify the different fungi present in the snuff sample.

·       To determine the microbial load of the powdered tobacco.

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