REDUCTION OF MICROBIAL LOAD OR KILLING THEM WITH NATURAL ALTERNATIVE BECAUSE OF SYNTHETIC CHEMICAL ADDITIVES.

ABSTRACT

The pervasive use of synthetic chemical additives in food preservation has raised concerns about their potential health hazards and environmental impact. In response, this study aimed to explore natural alternatives for reducing microbial load or eradicating microbes commonly found on vegetables and fruits due to inadequate disinfection practices. The primary objective was to mitigate microbial contamination using natural alternatives, particularly focusing on producing a vegetable sanitizer derived from plant extracts. Specific objectives included inhibiting common enteric pathogens like Escherichia coli (E.coli) and Staphylococcus aureus (S.aureus) associated with foodborne illnesses from contaminated produce.

Plant samples of Allium sativum (Garlic) and Ocimum gratissimum (Oregano) were collected and processed. Materials underwent thorough sterilization, and test organisms were obtained from laboratory cultures. Plant extracts were prepared through various methods, including ethanol and aqueous extractions. Disc diffusion and dilution methods were employed to assess antibacterial sensitivity and determine minimum inhibitory concentrations (MIC) of the extracts. The study revealed varying yields of extracts depending on the plant and extraction method employed. Both garlic and oregano extracts exhibited antimicrobial properties, with ethanol extraction yielding higher concentrations compared to aqueous extraction. The antimicrobial activity of the extracts was concentration-dependent, showing significant inhibition of S.aureus and E.coli. No resistance to the plant extracts was observed in the tested organisms.

The findings suggest the potential of garlic and oregano extracts as natural vegetable sanitizers, offering an alternative to synthetic chemical additives. Concentration-dependent antimicrobial activity underscores the importance of dosage in achieving desired effects. The study corroborates previous research indicating the effectiveness of plant extracts against pathogenic bacteria.

Garlic and oregano extracts demonstrate promising inhibitory effects against harmful enteric bacteria commonly found on unwashed produce. These natural herbs could serve as the basis for developing effective vegetable sanitizers, potentially reducing reliance on synthetic chemicals. Further research is warranted to explore the feasibility of applying these findings in real-world settings. The study underscores the potential of local herbs in combating microbial contamination in food. To advance this research, provision of laboratory equipment and public education initiatives highlighting the antimicrobial properties of natural herbs are recommended. Future studies should explore the efficacy of these natural alternatives in practical applications and assess their impact on microbial resistance patterns.

TABLE OF CONTENTS

CHAPTER ONE

INTRODUCTION

1.1 Background of the Study

1.2    Statement of Problems

1.3    Objectives

CHAPTER TWO

LITERATURE REVIEW

2.1 Natural Antimicrobial Products from Plants

2.2 Phytochemical Components of Plants

2.3 Herbs and Spices

2.4 Ocimium gratissimum (Scent Leave/ African Basil/ Nchuanwu)

2.4.1 Pharmacology of Extracts and Essential Oil of Ocimium gratissimum

2.5       Allium sativum (Garlic)

2.5.1   Antibacterial Activity of Allicin

2.5.2   Mechanism of Action of Allicin

2.6       Herbal Fumigant

2.6.1    Disinfection of Water by Herbs

CHAPTER THREE

MATERIALS AND METHOD

3.1       Collection of Plant Samples

3.1.2    Sterilization of Materials

3.1.3    Source of Test Organism:

3.2       Plant Extracts Preparation

3.2.1    Occimium gratissimum Extract

3.2.2    Disc Preparation

3.2.3    Antibacterial Sensitivity Testing Using Disc Diffusion Method

3.2.4    Dilution Method for Inhibitory Effect and Bactericidal Effect (Qualitative Test).

3.3.5    Determination of Minimum Inhibitory Concentration (MIC) Of the Crude Plant       

Extracts

CHAPTER FOUR

RESULTS

CHAPTER FIVE

DISCUSSION, CONCLUSION AND RECOMMENDATION

5.1 Discussion

5.2 Conclusion

5.3 Recommendation 

References

Appendix: Percentage Yield of Plant Extracts

LIST OF TABLE

Table 2.1       Sensitivity of Various Bacterial Species to Allicin

Table 4.1      Percentages of the Crude Extraction of Ocimium gratissimum and Allium Sativum

Table 4.2       Antimicrobial Activity of the Extracts with Diameter Zone of Inhibition (mm)

Table 4.3       Minimum Inhibitory Concentration (MIC) of Aqueous and Ethanol Extract of Ocimium gratissimum and Allium sativum Against Staphylococcus aureus and Escherichia coli

LIST OF FIGURE

Fig 1: Generation of Allicin in a Garlic Clove

CHAPTER ONE

INTRODUCTION

1.1       BACKGROUND OF THE STUDY

Plants such as fruits, vegetables, seeds, herbs and spices possess natural products which are chemical compounds or substances found in nature and they have pharmacological or biological activity (Koeh et al., 2005). Natural antimicrobials are been given more attention due to the increase concerns on chemical preservatives among consumers. Even though chemical preservatives are approved for human consumption by government agencies, many of these preservatives still threaten our health. Thus, the scientific communities have given more attention towards the potential antimicrobial activities of natural products. On the other hand, the increasing antibiotic resistance against chemical preservatives and sanitizer of some pathogen associated with food borne illness is in increasing rates (Sofos, 2008) and (Matthew et al., 2007).

In recent years, a large number of studies have been conducted searching the antimicrobial activity of natural products. Plants especially herbs and spices, are been given more attention. Now days, there are over 1340 plants with defined antimicrobial activities and over 30, 000 antimicrobial compounds have been isolated from plants (Tajkarimi et al., 2010). These plants and their compounds could be used as antimicrobial agents against food- borne pathogens like Escherichia coli, Staphylococcus aureus and Salmonella. They can also be used to control spoilage and pathogenic microorganisms in vegetable and other food products.

In addition, extraction methods, mechanism of action and factors influencing the antimicrobial activity are important of a natural sanitizer either for vegetables or in disinfection of inanimate objects.

EPA said that to sanitize is to make sanitary as by cleaning or disinfecting and to be sanitary is to be free from elements such as filth or pathogens that endanger health. Sanitizer are used to reduce, but not necessarily eliminate microorganisms from the in animate environment to levels considered safe as determined by public health codes or regulations, sanitizers includes food contact and nonfood contact products.

Sanitizing rinses for surfaces such a dishes and cooking utensils found in dairies, food –processing plants and eating and drinking establishments comprise the food contact sanitizers.

Spices have a long history of both culinary and medicinal uses (Tapsell and Swrahi 2006). Some species have been reported to have both bactericidal and bacteriostatic activities (Onwuliri and Wonang, 2005). They are generally recognized to be safe either because of their traditional use without any documented detrimental impact or because of dedicated toxicological studies (Smid and Gorris 1999). Some species are highly medicinal and have been used to sustain health and cure illness (Osunwole 1999). Some of the factors that determine their antimicrobial activities include concentration and composition of the spices, the type of microorganism, pH value, temperature and phenolic substances present in the food (Sagdic, 2003).   

Hence the use of safe antiseptics for hand wash purpose is necessary. Many of chemical antiseptics available in market are alcohol based and have some short comings or adverse effects, their frequent use can lead to skin irritation and also resistance among pathogens (Joshi et al., 2008). Number of herbs based products are being formulated and introduced into the market to overcome the problems. Enthapharmacologists, botanists, microbiologist and natural-products chemists are searching for photochemical and “leads” which could be developed into antimicrobials. Many plant extracts have been screened for their antimicrobial potential as it is very likely that these photochemical will find their way into arsenal of antimicrobials which may be used as disinfectants, antiseptics, dentifrices and chemotherapeutic agents (Marjorie, 1999).

1.4  STATEMENT OF PROBLEMS

Finding out the antibacterial activity of Ocimium grattissimum and Allium sativum as local herbs on Staphylococcus aureus and Escherichia coli which cause food borne diseases when vegetables contaminated by them are taking without proper sanitization and also because these organisms acquire resistance on the various chemical used in sanitization. This study now tries to develop a natural means of killing or reducing their load on food materials and surfaces and also to increase the shelf life of vegetables.   

1.5  OBJECTIVES

General objectives

i.        Reduction of microbial load or killing them with natural alternative because of synthetic chemical additives.

Specific Objectives

ii.      Producing a natural vegetable sanitizer

iii.    Inhibition of some enteric pathogenic (E.coli and S.aureus) that causes food-borne disease when infected vegetables and fruits are taken without proper disinfection.

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