Isolation And Identification Of Staphylococcus Aureus Associated In Contaminated “Okpa”  (Bambara Groundnut)  Voundzies Subterenea

 

 

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ABSTRACT

“Okpa” a processed food from bambara groundnut is a major source of dietary protein, and forms a major food in the Easter in part of Nigeria.
Isolation and identification of staphylococcus eureus associated with contaminated “Okpa” was were brought from Ogbete, Kenyatta and New Market different times of the day (8.00am, 12noon, 4.00pm).
Isolate from the study also include salmonella species Clostridum botulmm Bacillius Cereus Campylobacter Jejum, Aenomoanas hydrophilia.
This study has shown that temperature is a very important determinant of microbial physiology and their availability in media. The importance of hygiene cannot be over emphasized.

 

 

TABLE OF CONTENT

CHAPTER ONE
1.0 Introduction 1
1.1 Aim and Objectives 5
1.2 Statement of Problems 6
1.3 Hypothesis 6
1.4 Limitations 6
1.5 Justification 7

CHAPTER TWO
2.0 Literature Review 8
2.1 Legumes and Food 8

CHAPTER THREE:
Methodology 16
3.0 Method of Sample Collection and Material 16
3.1 Materials 18

CHAPTER FOUR
4.0 Result 25
4.1 Mean Bacterial Count on “Okpa” in different
media at different time of the day 27

CHAPTER FIVE
5.0 Discussions 30
5.1 Recommendation/Conclusion 32
References 34

 

LIST OF TABLES

TABLE I: Mean bacterial count of “okpa” in different media at different time of the day.

TABLE II: Identification of Bacteria Isolated from “Okpa”
TABLE III: Flow chart showing production of “okpa” from Bambara groundnut.

 

 

 

CHAPTER ONE

1.0 INTRODUCTION
The legumes used as food can be divided into two groups, pulses and oil seed pulses are the dried seed of cultivated legumes while oil seeds are those legumes cultivated primarily for the oil content which may be extracted by pressing or by solvent extraction, the residue being a high protein cake. The nutritive properties of pulses resemble. Pulses are rich in lysine, in which many cereals are deficient legumes contain a considerable amount of carbohydrate (60% on the average) mainly starch, which is generally well digested and absorbed.
(Report of an Adhoc panel of the Advisory committee on Technology Innovation 1984) legumes generally contain low fat.

 

Pulses are good sources of B – group of vitamins. Greater part of these vitamins present in the harvested seeds are actually consumed unlike in cercals (rice and wheat) where there are losses that may arise during melling and cooking of rice. Pulses have therefore a well deserved reputation as a food will protect consumers against beri – beri (Davidson, 1975).
Bambara groundnut is one of the most popular pulses among Africans. It produces a nutritions food and is cultivated throughout Africa.

 

“Okpa is a processed food from Bambara groundnut with its biological name as voandzers subterenea by (Onyekwere, 1977). It forms a major sources of dietary protein food in the Eastern part of Nigeria among the Ibo’s especially those from Enugu State Bambara groundnut is produced in pods and its seasonal.
In some of Eastern States of Nigeria “Okpa” is considered as second to bread. (Report of an Adhoc panel of the Advisory Committee on Technology Innovation 1984). The Bambara seeds are also known as to have different names among the different ethnic groups in Nigeria. The Ibo’s call it “Okpa”, the Hausa’s call it “juijiya”, while the Yoruba’s call it “Epivour”.

 

In Eastern parts of the Nigeria, the flour from the seed is prepared and eaten as “Okpa”. This is gotten by mulling dry mature seed, mux with hot water and condiments to form paste. The resulting paste is wrapped in non absorbent leaves and steamed to solidify okpa, is a favourbale food especially among those who are familiar with it.
Staphylococcus aureus is a spherical bacterium (coccus) which on microscopic examination appears in pairs, short claims, or bunched, orape – life chisters.

 

The organism are gram – positive. Some strains are capable of producing a highly heat stable protein toxin that cause illness in humans. The on set of symptoms in staphylococcus food poisoning is usually rapid and in many eases acute, depending on individual susceptibility to toxin, the amount of contaminated ingested and the general health of victim. The most common symptoms are nausea, vomiting, abdominal grapping, and prostration some individuals may not always demonstrate all the symptoms associated with the illness like sever cases, headache, muscles cramping, and transient changes in blood pressure and pulse rate may occur.

 

According to Hobbs and Robert (1993) package food gets contaminated with micro organism during the processing period. These includes the water for preparation, cooking utensils, the milking machine, the food handlers and packaging materials, if not properly washed.

 

This Staphylococcus aureus found in “Okpa” enter the host cell, invade and multiply in or on the body tissue of the host. It was early in the 20th century that bacteria was first recognized as against of food – poisoning and food borne disease (Hobbs and Roberts, 1993). This bacteria includes:- Clostrdum Perfringes salmonella specie, Bacillus cereus campylobacter jejuni are known as food poisoning agent common in many countries Glostridium Botulmum is a hazard in Northern countries and Japan. Certan types of Escherichia Coli are known to cause diarrhoea in adult, which is often referred to as travelers diarrhoea. Streptococli are occasionally incriminated in food poisoning other organisms, particularly the common gram negative bacilli have been reported from times to time (Hobbs and Roberts, 1993).

 

The aim of this work therefore is to determine the bacteria, Staphylococcus aureus found in contaminated “okpa” at different time of the day (ie determine) when “okpa” like every other food is no longer good for human consumption. It is hoped that result from this investigation will provide useful information on the bacteria contamination of “okpa” and lead to improvement in the method of “okpa”

 

 

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Comparative Analysis Of Pipeborne Water And Other Sources Of Water Within Enugu Metropolis (Emene Locality)

 

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ABSTRACT

 

Comparative analysis of water was carried out on well water, stream and pipe born water obviated within Emene locality of Enugu state the sample were subjected to some analytical tests final like presumptive tests, comfirmative test final like presumptive tests, comfirmative test, completed test, indole test and plate count for the organism that develop on the culture, the result show that well water and stream were found to contain pathogenic organism like salmonella, Shigella and staphylococcus aureus as well as numerous viable counts, the cloform or the indicator organisms Escherichia coli the family of enterbactercteriaceae were isolated too form well water the pipe borne water a sample contained staphylococcus aureus and have lowtotal viable counts.

 

 

 

 

 

TABLE OF CONTENTS

 CHAPTER ONE: INTRODUCTION

1.1     Aims and objective

1.2     Statement of problem

1.3     Hypothesis

1.4     Significance of study

1.5     Limitations

• CHAPTER TWO LITERATURE REVIEW
  • water analysis

 

3.0     CHAPTER THREE: MATERIALS AND METHOD  

3.1     Sample collection

3.2     Preparation of media

3.3     Sterilization of materials

3.4     Multiple tubes technique

3.5     Presumptive test

3.6     Confirmed test

3.7     Completed test

3.8     Test for indole

3.9     Methyl red test

3.10 Oxidase test

3.1.1 catalse test

3.12 plate counts techniques

3.13   preparation of single and double strength media

3.14 dilution technique in most problem number     `

3.15 Inoculation of media with water samoltes

 

4.0 CHAPTER FOUR: RESULTS

GRAM STAIN TECHNIQUE                                                  

4.1 Colony counts

4.2 Most problem number (mpn) and pH result.

• Chapter five. Discussion an

5.1     Discussion

5.2     Recommendation

5.3     Conclusion

6.0     References

Appendix

LIST OF TABLE S

 

Table I comparative guality of ground water and surface water

12

Table II colony count result                                                                        25

Table III MPN and P^H result                                                                       25

Table IV the MPN of cliform     in the dilution                                                 27

Table V the physiological biochemical of the isolated organisms               28

 

 

 

 

 

 

 

 

 

 

LIST OF FIGURES

 

Fig I culture plate picture                                                       23

Fig II isolated micro organisms and shape                           24

 

 

 

 

CHAPTER ONE

 

INTRODUCTION

Following the theory of creation, it is clear that water is as old as man. From time immemorial, man has restorted to the use of this unique commodity for domestic and other purposes.

Most of our water supplies are from surface water which include: rivers, streams, lakes, oceans and seas and there water bodies are likely to be polluted with domestic and Industrial as well as agriculture waste, As populations increase, the problem become more serious and as such, water can endanger the health and life of human beings because when polluted by fecal materials it becomes potential carrier of pathogenic organism (Carpenter 1977).

Water, is of course, absolutely essential to life, not only human life but all life, animal and vegetable. Most of the biochemical reaction that occur in metabolism and growth of living cells involved water, and all take places in water. (Camp etal,1974).

Man uses water not only for drinking purposes but also for bathing, washing, laundering, heating, air conditioning, agriculture, stock raising and gardens, Industrial processes and cooling water power and steam power, fire protection, fishing, swimming and wild life propagation and navigation.

Natural water contain not only then natural flora but also micro – organisms from soil and possible from animals or sewage. Surface waters in streams or pools and stored waters in lakes and large ponds vary considerably in microbial content. (frazier 1978) water is broadly divided into three types viz., surface water which include: streams, rivers, lakes sear, and oceans Kelman et al (1957)

The ground water, well borehle, many people have defined the ground water in different ways: ground water is subter water that occurs where all pores in the soil or rock counting materials are saturated (pelezer et al 1992).

The atmospheric water, which includes rain, falls. All water bodies consist of a variety of bacterial and other microorganisms like the Algaes fungi, which inhabit these natural water bodies. Some of these micro –organisms are indigenous to thus natural water while others are transient, entering the water from external environment (Pelezar and Reg, 1997).

The generality of bacteria are mostly commonly found ordinarily in fresh water some of which include: pseudomonas, Archacbacter, and vibrio these are gram negative, the gram-positive bacterial which are found in water include: micrococcus Archacbacter and actinomycentes. Gebharal (1975) Tap water, as one of the water sources is mostly used domestically, it is observed that tap change sometime the water tap will be clear this calls for load, in order to be sure of its portability. Bonde (1977)

The increase in drinking water from different sources especially in Enugu state has made necessary to investigate the microbial content of water. Water is a potential carried of pathogenic organisms that can endage human life. Most of drinking water sources are often contaminated with different pollutants like faces, animal and plant wastes, making such water unfit for drinking if not treated. The pollution of water with pathogenic organisms and other pollutants can only be detected by carrying out microbiological assessment of such water. Most human disease such as typhoid paratyphoid cholera, arnboebiasis, Trichinosis, gastroenteritis, sanonall shigellosis, diphtheria, giadia, dracunculus etc are know to be water borne disease. Ewington et al (1971).

Water born disease are those disease which have water as their vehicle of transmission these disease are capable of destroying a whole community if not checked. Therefore, the quickest ways to prevent out break of these disease and to determine the portability of such water sources is to determine the microbial load or content if the microbial content is nor within acceptable limit, such water sources should be condemned immediately. Fair et al (1970)

 

 

 

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The Antibacterial Activity Of Three Types Of Medicated Soaps On Starhyrococcus Aureus Form Would Infections

 

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ABSTRACT

The antibacterial activity of three type of medicate soap on staphylococcus aureus isolated from wound infections was conducted. Fifty individual with would infection within the age range of 9-73 years were sampled. Swabsticks were used to collect specimens from wound infection. each swabstick was streaked separately over plates of nutrient and macconkey agar and later incubated at 370c. out of the fifty (50) individuals sampled 25(50%) were mostly infected with staphylococcus aureus followed by pseudomonas acruginqu (22%) staphylococcus epidermis (10%) least by Exchericha coli b(12%) The organisms occur within the age range 9-13 years with 8 (16%) followed by 14-18 years 4(8%) next by the age range of 19-23 years 3(6%) followed by 24-28 years with 2(4%) followed 24-28 years with 2(4%) and the heart in the age range are 29-33,34-38,39-43,44-48,49-53,54-58 64-68 with 1(2%) each while there was no isolation in the age range 59-63. The study also revealed that all the staphylococcus aureus isolated were sensitive to the three medicated soap. The means of inhibition was highest in Aleppo medicated soap with 12.92 mm followed by temperate medicated soap with 11.2mm and least in carat medicated soap with 10.56 mm. Also the staphylococcus aureus isolated were also sensitive to the commercial autibutics used as couplers the means zones of inhibition was highest in ampiclox with 12.8 mm followed by gentamyain with 12.04 mm and the heart in penicillin with 10.4 mm. there was a close relationship in the zenes of inhibition between the three medicated soap and the commercial antibiotics used as coconutrels. The result showned that medicated soaps were effective against staphylococcus aureus involved in would infection in humans and are comparable to commercial antibiotics. There fore medicated soap can be used in cleaning the skin particularly in the affected part of the skin during the time one has hurt or cut on the skin.
TABLE OF CONTENTS

CHAPTER ONE
1.0 Introduction
1.1 Aims and objectives
1.2 Statement of the problem
1.3 Hypothesis
1.4 Justification of the study
1.5 Limitation Of The Study
CHAPTER TWO
2.0 Literature review
2.1 Antibacterial activity of soap
2.2 Medical relevance of antimicrobial soap
2.3 Normal skin microbiota and soap

CHAPTER THREE
3.0 Material and methods
3.1 Materials
3.2 sterilization
3.2.2 Collection of soap sample
3.2.3 Preparation of culture media
3.2.4 Collection of bacterial organisms from would infection
3.2.5 Microbial count, grams stain and microscopic examination
3.2.6 Biochemical test for identification of isolates

CHAPTER FOUR
Result

CHAPTER FIVE
Discussion

CHAPTER SIX
Conclusion and recommendation
References
Appendix
LIST OF TABLE

Age distribution number sampled as well as percentages with staphylococcus aureus and other organisms
Table 2
Age distribution as well as percentage of each isolated organisms
Table 3
Preliminary identification of bacterial isolates
Table 4
Biochemical test carried out for identification of bacterial isolates
Table 5
Sensitivity test using three medicated soap and three antibiotics indicating their zones of inhibition.
CHAPTER ONE

INTRODUCTION
According to pelage et al (1986) antibacterial activity is the ability to either destroy bacteria or inhibit their growth. This is significant with respects to the human body in preventing sepsis and skin infections. Also Derland (1981) states that medicated soap have the germicidal substance like chloroxyhlenol potassium mercuric iodide, trichlorocarbanlide etc. incorporated into them, in order to enhance their antibacterial activity. These germicide substance are normally added in a specified amount and percentage of the substance used are always stated on the soapcase or inside the lesflet which certains the information on how to use the soap for various purposes.
Anon (1964) states that soap may be defined as a chemical compound resulting from the interaction of fathy acids oil and caustic soda (alkali) possesing the characteristic soap like properties of detergents, surface tension lowering wetting and emulsifying power and gel formation. All fatty oils and fats. Are mixtures of glycoside compounds (erters) of trihydric alcohol, glycosides and some fatty acids such as plasmatic acid the chemistry of soap manufacturing may be expressed the following equation which is know as saponification (Anon (1964).
HzCocoG5H31)3 + Na 0H Naoco15H31 (zH31 + c3H5(oH)3
Palliation caustic sodium palurtate glycerion
Soda
Amon (1964) further explain that a bactericidal soap cernirts existentially of the following 0.1 to 3% weight based on the total weight of the soap.
0 to 10% by weight based on the total weight of the soap in a compound of formular
R –Co-N1-CH2C00H
CH3

Here R represents an alkyl or alkenye group of 8 to carbon atoms.
According to Johnson (1978) antibacterial soap may include 2,4,4,trichloro 2,2 hydroxydiphenyl in an amount 0.05 to 5% by weight and at least one phosphorus oxyacid. Phosphorus oxyacid salt phosphoric ester is represented by
(R1(oCH,CH2)
R2

Where R1 in (8-20 alkyl, R2 is hydrogen or methyl and is (1-10). R1 is the same group as R2 or H alkali metal and n is H or alkali metal. Such a soap has a wide range of antibacterial activities and marked resistance of discoloration upon exposure to sunlight (Kaw,1981) another invention relates to an autimicrobial composition consisting of ammoniated zinc sulphate optionally formulated into sufficient formulation and toilet bars. Medicated soap incorporate in their composition germicidal agent which include hexachlorophene mercuric which trichoran, trichlorocarbaucide .
Sykes (1958) described sterilization as the complete destruction of all living matters. In medical sense, it is often used in a restricted sense to refer to the destruction of pathogenic organisms only.

According to William (1979) wound is defined as disruption of cellular and anatomic continuity while its healing is the restoration of continuity. That biological procen can only be accomplished by regeneration, cell proliferation and collage production which can be alleviated washing the would surface especially with medicated soap which due to its concert of pherolic compound help in keeping off organisms like staphylococcus aureus escherichia coli and Pseudomonas aeruginosa always from the would to a certain stage.

Wound can also result when the operative barrier of the skin is breached by traumatic invasion or whether it is caused by trauma or internationally by surgery. The open area is susceptible to microbial invasion and once a would has become infected pus form in the injured area resulting to wound abbess.

Baker et al decried antisepsis as the most convenient way of preventing infection usually by inhibiting the growth of bacteria. And most disinfectant when suitably diluted have the artixeptic action. Different method are employed for the destruction of bacteria or for getting rid of them and those method can be conveniently divided into chemical physical and mechanical methods.

Baker et al (1985) explained that chemical agent function as a sterilizing agent by the following lethal mechanisms
1. Disruption of the cell membranes
2. Interfering with orgymatic systems of the organisms emzyme poison)
3. Co-aqulation of protein
4. Oxidation

 

1.1 AIMS AND OBJECTIVES OF THE STUDY
1. To isolate staphylococcus aureus from would infection
2. To determine the artibacterial activity of three medicated soap on staphylococcus aureus isolated from wound infection.

 

 

1.2 STATEMENT OF PROBLEM
Since Dorland (1981) stated that medicated soaps have germicidal substances like chloroxylenol pothaium, mercuric widide trichlorocarberihide etc incorporated into them in order to greatly their antibacterial activity. It is…

 

 

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