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IDENTIFICATION OF FUNGI AND DETECTION OF MYCOTOXINS ASSOCIATED WITH STORED LOCUS BEANS PURCHASED FROM YANKURA MARKET

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BY

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--/H2013/01430
DEPARTMENT OF ----
SCHOOL OF ---
INSTITUTE OF ---



APPROVAL PAGE

This is to certify that the research work,"identification of fungi and detection of mycotoxins associated with stored locus beans purchased from yankura market"by ---, Reg. No. --/H2007/01430 submitted in partial fulfillment of the requirement award of a Higher National Diploma on --- has been approved.

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Supervisor                                                  Head of Department.
Signature……………….                           Signature……………….        

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DEDICATION
This project is dedicated to Almighty God for his protection, kindness, strength over my life throughout the period and also to my --- for his financial support and moral care towards me.Also to my mentor --- for her academic advice she often gives to me. May Almighty God shield them from the peril of this world and bless their entire endeavour Amen.



ACKNOWLEDGEMENT

The successful completion of this project work could not have been a reality without the encouragement of my --- and other people. My immensely appreciation goes to my humble and able supervisor mr. --- for his kindness in supervising this project.
My warmest gratitude goes to my parents for their moral, spiritual and financial support throughout my study in this institution.
My appreciation goes to some of my lecturers among whom are Mr. ---, and Dr. ---. I also recognize the support of some of the staff of --- among whom are: The General Manager, Deputy General manager, the internal Auditor Mr. --- and the ---. Finally, my appreciation goes to my elder sister ---, my lovely friends mercy ---, ---, --- and many others who were quite helpful.


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ABSTRACT

Mycotoxins are toxic secondary metabolites produced by various molds and fungi. While they are more commonly associated with crops such as grains (locust beans). Mycotoxins are secondary metabolites generated by several species of fungus that have a negative impact on food quality and are dangerous for both people and animals. Aflatoxins (AF), Fumonisins (FUM), Deoxynivalenol (DON), Ochratoxin A (OTA), Zearalenone (ZEA), Patulin (PAT), and Citrinin (CIT) are the most prominent and commercially relevant mycotoxins TLC has become a remarkably efficient, fast, and, in most circumstances, inexpensive separation technique in mycotoxicology. In this study, mycotoxins from various fungi, including Aspergillus species, Penicillium species, Fusarium species, Mucor species, Nocardia species, Trichoderma species, Curvularia species, Bipolaris species, Rhizopus species, and Alternaria species, are screened using TLC (Thin layer chromatography) analysis method, an easy physicochemical experiment, to determine whether they are present in stored locust beans. The extraction of mycotoxins used a variety of solvent systems. The study focused on the detection of mycotoxins in stored locust beans samples using Thin Layer Chromatography (TLC) technique. Fungal species, including Aspergillus sp, Penicillium sp, Fusarium sp, Mucor sp, Nocardia sp, Trichoderma sp, Curvularia sp, Bipolaris sp, Rhizopus sp, and Alternaria sp, were screened for mycotoxins. The TLC plates were visualized under visible light and UV light to identify the presence of mycotoxins. The study identifies fungi and detects mycotoxin contamination associated with stored locust beans purchased from yankura market using Thin Layer chromatography.Findings of this study can help create methods for preventing the formation of mould and extending the shelf life of locust beans.

TABLE OF CONTENTS
COVER PAGE
TITLE PAGE
APPROVAL PAGE
DEDICATION
ACKNOWLEDGEMENT
ABSTRACT
CHAPTER ONE
1.0      INTRODUCTION
1.1      BACKGROUND OF THE PROJECT

    1. STATEMENT OF PROBLEM
    2. AIM AND OBJECTIVE OF THE STUDY
    3. HYPOTHESIS
    4. SIGNIFICANCE OF THE STUDY
    5. LIMITATION OF THE STUDY
    6. SCOPE OF THE STUDY
    7. METHODOLOGY
    8. STRUCTURE OF THE STUDY

CHAPTER TWO
LITERATURE REVIEW

    1. OVERVIEW OF LOCUST BEAN
    2. BENEFIT OF LOCUST BEAN
    3. SOURCES OF MICROORGANISMS IN FOODS
    4. MICROBIAL ACTIVITIES IN FOODS
    5. CONTROL OF MICROORGANISMS IN FOODS
    6. APPROACHES TO MICROBIOLOGICAL CONTROL IN FOODS

CHAPTER THREE
3.0      RESEARCH METHODOLOGY

    1. INTRODUCTION
    2. STUDY AREA AND SAMPLE SOURCE
    3. ISOLATION AND IDENTIFICATION OF FUNGI
    4. MASS PRODUCTION AND EXTRACTION OF FUNGI

CHAPTER FOUR
4.0      RESULT AND DISCUSSION
4.1      Isolation and Identification of Fungi
4.2      Submerged Fermentation and Extraction Method
4.3      Mycotoxins Detection by TLC Method (Thin Layer Chromatography)
CHAPTER FIVE

    1. CONCLUSION AND RECOMMENDATION
    2. REFERENCES

  

CHAPTER ONE
1.0                                                        INTRODUCTION
1.1                                           BACKGROUND OF THE STUDY
Locus bean is a popular condiment used as taste and flavour enhancer in soup and dishes in Africa. IARC (2013) stated that fermented locust bean seed is commonly consumed in Ghana, Nigeria, Sierra- Leone and Togo. In Nigeria it is called iru in Yoruba, dawadawa in Hausa and ogiri `igala in Igbo. It is also referred to as kinda in Sierra- Leone and kpalugu in Ghana. When locust beans is stored, it means that it is preserved. Preservation and preservatives are designed to inhibit/control the activities of spoilage causing organisms in food, a process also referred to as sanitization. Spoilage causing organisms due to their growth and metabolic activities produce by-products, which change the texture, taste, flavor and the aroma of the food. Preservatives by their nature are intended to keep the food devoid of these changes. Preservatives act on both Gram positive and Gram negative food spoilers (Daou et al., 2021). The general idea of products having preservatives is to increase shelf life and prevent items from spoiling. The more shelf life a            product has, the more marketable it becomes. The use of preservatives makes products stay fresher, longer and give more time for products to be used (Richard, 2017). Preservation and preservatives are designed to inhibit/control the activities of spoilage causing organisms in food, a process also referred to as sanitization. Spoilage causing organisms due to their growth and metabolic activities produce by-products, which change the texture, taste, flavor and the aroma of the food. Preservatives by their nature are intended to keep the food devoid of these changes. Preservatives act on both Gram positive and Gram negative food spoilers (Awuchi et al., 2020). The general idea of products having preservatives is to increase shelf life and prevent items from spoiling.
The more shelf life a product has, the more marketable it becomes. The use of preservatives makes products stay fresher, longer and give more time for products to be used (Richard, 2017).
Preservation and preservatives are designed to inhibit/control the activities of spoilage causing organisms in food, a process also referred to as sanitization. Spoilage causing organisms due to their growth and metabolic activities produce by-products, which change the texture, taste, flavor and the aroma of the food. Preservatives by their nature are intended to keep the food devoid of these changes. Preservatives act on both Gram positive and Gram negative food spoilers (Awuchi et al., 2020). The general idea of products having preservatives is to increase shelf life and prevent items from spoiling. The more shelf life a product has, the more marketable it becomes. The use of preservatives makes products stay fresher, longer and give more time for products to be used. Shelf life is the period of time during which the food product remain safe, retain desired microbiology, physical, chemical and sensory characteristics (Ademola et al., (2013). The aim of the research is to determine microbiological quality of fermented African locust bean cake stored under ginger extract and ascorbic acid treatment.
In this study various bacterial and fungal species associated with some locally fermented products in locust beans were isolated and identified. Moreover investigation on microbiological hazards associated with fermented locust beans was also intensified to safeguard the production of this food condiment meant for human consumption.

1.2                                          STATEMENT OF PROBLEM

Safe food is a basic human right despite that many food condiments are frequently contaminated with naturally occurring pathogenic microorganisms. Such pathogens cannot be detected organoleptically (seen, smelled or tasted), but can cause disease of varying severity, including death specially if the way they are conserved during exposition for sales provides conditions for those microorganisms to grow and reach considerable levels of contamination (Adeyeye et al., 2016).

The global incidence of food borne illnesses is difficult to estimate but it has been reported that in 2000 alone 2.1 million people died from diarrhoeal diseases. A great proportion of these cases can be attributed to contamination of fungi and mycotoxins. This study identifies fungi and detects mycotoxin contamination associated with stored locust beans purchased from yankura market using Thin Layer chromatography.

1.3                          AIM AND OBJECTIVES OF THE STUDY
This study aimed at identifying fungi and detection of mycotoxin contamination associated with stored locust beans purchased from yankura market using Thin Layer chromatography. The objectives of the study are:

  1. To determine the physiochemical parameters of the stored locus beans
  2. To isolate and identify fungi from the stored locus beans
  3. To detect the presence of mycotoxins associated with the stored locus beans using Thin Layer chromatography

1.3                                                         HYPOTHESIS
HO:     fungi and mycotoxins are found in stored locust beans
Hg       = fungi and mycotoxins are not found in stored locust beans

1.4                                           SIGNIFICANCE OF THE STUDY
This study will be useful to consumers of food. The outcome of this research will guide users environment on the best method of preserving locust beans in other to avoid fungi and mycotoxins. The research work also will add to the literature and information existing about our environment

1.5                             LIMITATION OF THE STUDY
There are many or different type of microorganism that are found in different food item, the focus of this work  is made only on fungi and mycotoxins found in stored locust beans

1.6                                                   SCOPE OF THE STUDY
Fungi and mycotoxins are all around us. This study allows students to collect samples from the stored locust beans and grow the microorganisms so that students can visualize the microorganisms.  This teaches students how to fungi and mycotoxins grow in stored locust beans. The approach used in this work will enable us to evaluate the mycotoxicological risk of stored locust beans, to minimize economic losses and reduce the hazard to animal and human health.
1.7                                              RESEARCH METHODOLOGY
In the course of carrying this study, numerous sources were used which most of them are by visiting libraries, consulting journal and newspapers and online research which Google was the major source that was used.

1.8                                               STRUCTURE OF THE STUDY
The work is organized as follows: chapter one discuss the introductory part of the work,   chapter two presents the literature review of the related works, chapter three describes methods used, chapter four discusses the result analysis, chapter five is on summary of findings, conclusion and recommendation.

CHAPTER FIVE

5.1                                    Conclusion and Recommendation

Mycotoxins are harmful compounds made by certain moulds (fungi) that may infect a range of food items, including baked products. These toxins pose health risks to humans and animals when ingested in sufficient quantities. While mycotoxin contamination is more commonly associated with grains, nuts, and other staple foods, bakery products can also be affected if the raw ingredients used in their production are contaminated. The present study focuses on the detection of mycotoxins in bakery food products using the Thin Layer Chromatography (TLC) technique. The study involved the isolation and identification of various fungal species from stored locust beans samples, followed by the extraction of mycotoxins using different solvent systems. The mycotoxins were then analysed using TLC plates and visualized under UV light. The results showed the presence of mycotoxins in certain fungal species, indicating their ability to produce toxins. The findings of this research can contribute to the development of strategies to prevent mold growth and ensure the safety of locust beans.
It's important to note that mycotoxin contamination in locust beans is relatively rare compared to other food categories, but it is still a concern for food safety. Both food producers and regulatory agencies work to ensure that food items and other food products meet safety standards and pose minimal health risks to consumers.

 


CHAPTER TWO: The chapter one of this work has been displayed above. The complete chapter two of"identification of fungi and detection of mycotoxins associated with stored locus beans purchased from yankura market"is also available. Order full work to download. Chapter two of"identification of fungi and detection of mycotoxins associated with stored locus beans purchased from yankura market"consists of the literature review. In this chapter all the related work on"design and construction of an arduino based public address system"was reviewed.

CHAPTER THREE: The complete chapter three of"identification of fungi and detection of mycotoxins associated with stored locus beans purchased from yankura market"is available. Order full work to download. Chapter three of"identification of fungi and detection of mycotoxins associated with stored locus beans purchased from yankura market"consists of the methodology. In this chapter all the method used in carrying out this work was discussed.

CHAPTER FOUR: The complete chapter four of"identification of fungi and detection of mycotoxins associated with stored locus beans purchased from yankura market"is available. Order full work to download. Chapter four of"identification of fungi and detection of mycotoxins associated with stored locus beans purchased from yankura market"consists of all the test conducted during the work and the result gotten after the whole work

CHAPTER FIVE: The complete chapter five of"identification of fungi and detection of mycotoxins associated with stored locus beans purchased from yankura market"is available. Order full work to download. Chapter five of"identification of fungi and detection of mycotoxins associated with stored locus beans purchased from yankura market"consist of conclusion, recommendation and references.

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