Enumeration And Identification Of Bacteria On Used Handkerchiefs In Males

Project and Seminar Material for Biochemistry

Enumeration And Identification Of Bacteria On Used Handkerchiefs In Males


Abstract


This study was carried out on the enumeration and identification of bacteria on used handkerchiefs in males. The study was carried out in Makurdi town. Study was conducted on four samples used male handkerchiefs. (Sample A, Sample B, Sample C, Sample D Handkerchiefs). Each sample was collected in isolation in sterile polythene bags to avoid cross contamination.The identification of bacteria was based on their morphological and biochemical characteristics using procedure described by Cheesbrough, 2000.Microorganisms were gram stained,Air dried and viewed under microscope under x10 objectives.Growth on the sabouraud dextrose agar plates were observed for colour, pigment production, size of growth opaquecity.

Data was analyzed using the descriptive statistic SPSS (version 20). The sample in this study were contaminated with the following contaminant for bacteria they were all colonized by one type of bacteria, Bacillus substilis. Fungal includes Candida albican, Candida tropicalis, Penicilliumspp, Rhodotorularubra, Trichophytonrubrum. This study has shown that used male handkerchiefsare colonized with one type of bacterial, Bacillus substilis. From this study, it is concluded that Sample B and Sample C have the highest percentage in fungal and all the handkerchiefs have the same percentages in bacteria isolates.


Table Of Contents


Preliminary Page(s)

  • Title page
  • Certification page
  • Dedication
  • Acknowledgement
  • Abstract
  • Table of content

Chapter One:

Introduction

  • 1.1 Background of the study
  • 1.2 Identification of Gram-Positive Cocci
  • 1.3 Statement of problem
  • 1.4 Objectives of the study
  • 1.5 Significance / Justification of the study

Chapter Two:

Literature Review

  • 2.1 Overview of bacteria
  • 2.2 Origin and early evolution
  • 2.3 Morphology
  • 2.4 Bacterial isolation and identification
  • 2.5 Human microbiome

Chapter Three:

Materials And Methods

  • 3.1 Area of Study
  • 3.2 Sample collection
  • 3.3 Media Preparation
  • 3.4 Inoculation/culturing
  • 3.5 Identification of Bacteria
  • 3.6 Morphological Identification
  • 3.7 Gram Reaction
  • 3.8 Coagulase Test
  • 3.9 Motility Test
  • 3.10 Oxidase Test
  • 3.11 Catalase Test
  • 3.12 Identification of Fungal
  • 3.13 Germ Tube Test
  • 3.14 Lactophenol (Cotton blue test)
  • 3.15 Statistical Analysis

Chapter Four:

Result And Discussion

  • 4.1 Results
  • 4.2 Discussion

Chapter Five:

Conclusion And Recommendations

  • 5.1 Conclusion
  • 5.2 Recommendation
  • References

Chapter One


Introduction

1.1 Background Of The Study

Micro-organisms are ubiquitous and are found in almost every area around human bodies. Some are specifically found in certain regions of the body as a normal flora where they live as commensals with man. This association is important in protecting the body against other infectious diseases.

Each area of the body surface acquires a characteristic flora of organisms well adapted to growth at that particular environment. These residents (normal flora) tend to suppress the intruders either by competition for space and food supply or by production of metabolites that are antagonistic to the survival of the intruder.

These residents could be dislodged from their environment when sneezing, coughing, belching, yawning or could be destroyed by regular use of antiseptic soaps or creams on the body surfaces.

Handkerchiefs often used in males for wiping face, closing of the mouth and nose when expressing these reflex activities, therefore constitute an abode for bacteria. Furthermore, bacteria found in handkerchiefs could differ from one individual to another as the bacteria found could be a reflective of the environment and pathological conditions of the individual using the handkerchief. For instance, individual with upper respiratory tract infection are likely to dislodge strains of pathogenic microbes along sides with the normal flora in these regions.

Enumeration of bacteria on used handkerchief in males can be done using microscopic cell count and viable cell counting. Microscopic counts can be done on either samples dried on slides or samples in liquid. A viable cell counting is the one that is able to divide and form offspring.

Viable cell counting is also called plate count and there are at least two ways of performing plate count: the spread plate and pour plate method.

In spread plate method, a volume of appropriately diluted culture is spread over the surface of an agar plate using a sterile glass spreader. The plate is then incubated until colonies appear, and the number of colonies formed are counted.

In pour plate method, a known volume of culture is pipetted in a sterile petridish plate. Molten agar medium is then added and mix well by gentle swirling of the plate on the bench top. Because the sample is mixed with molten agar medium, the bacteria to be counted must be able to withstand brief temperature exposure to the temperature of the molten agar (45 – 50oC). Here, the colonies formed are counted throughout the plate and not just on the agar surface as in the spread plate method.

In identifying bacteria, the morphological and biochemical characteristics of the bacteria, are evaluated. The appearance and the microscopic description of the bacteria are examined with the aid of a light compound microscope.

From the growth of the bacteria (pure culture), the specimen to be viewed under the microscope can be prepared as a smear or as a wet mount. A stain is used to contrast the specimen from the background. This strain could be basic or acid stain.Basic stain, example methylene blue and crystal violet, are cationic and have a positive charge. They are ideal for staining chromosomes and the cell membrane of the bacterial. The acid stains are anionic and have a negative charge, and are used to stain cytoplasmic material and organelles or inclusions. Common examples are eosin and picric acid.

There are two types of stains simple and differential. A simple stain has a single basic dye that is used to show shapes of cells and structures within a cell while a differential stain consists of two or more dyes and is used in the procedures to identify bacterial. One of the most commonly used differential stain the gram stain. Gram positive bacteria stain purple while Gram negative bacteria stain pink. Other biochemical tests like Indole test, Urease test, Catalase reaction, Oxidase reaction etc will help in further characterization of the bacteria identified from the handkerchief.


1.2 Identification of Gram-Positive Cocci

Gram-positive cocci can be identified by the growth on blood and chocolate agar and the catalase test. The catalase test is used to differentiate those bacteria that produce the enzymes catalase, such as staphylococcus from non-catalase producing bacteria such as streptococci. For catalase positive, gram positive cocci, coagulase test could be used to further differentiate them. Staphylococcus aureus is coagulase positive whereas Staphylococcus epidermidis is negative to coagulase test. Streptococci are catalase-negative gram-positive cocci and are further classified on the basis of their type of haemolysis, A (partial) and B (complete) haemolysis on blood agar, Alpha-haemolytic streptococci, streptococcus viridans and Streptococcus pneumonia can be differentiated by the optochin disc susceptibility test.

The B-haemolytic streptococci are group according to the lancefield classification.
Identification of Gram-Negative Cocci Members of this group can be identified using fermentation patterns. In addition, catalase and oxidase tests can also be performed. It can further be identified by growing them on Thayer-Martin medium and nutrient agar.


1.3 Statement of problem

Bacteria are living things that have only one cell. Under a microscope, they look like balls, rods, or spirals. They are so small that a line of 1,000 could fit across a pencil eraser. Most bacteria won’t hurt you – less than 1 percent of the different types make people sick. Many are helpful. Some bacteria help to digest food, destroy disease-causing cells, and give the body needed vitamins. Bacteria are also used in making healthy foods like yogurt and cheese. But infectious bacteria can make you ill. They reproduce quickly in your body. Many give off chemicals called toxins, which can damage tissue and make you sick. Examples of bacteria that cause infections include Streptococcus, Staphylococcus, and E. coli.

These residents could be dislodged from their environment when sneezing, coughing, belching, yawning or could be destroyed by regular use of antiseptic soaps or creams on the body surfaces. Handkerchiefs often used in males for wiping face, closing of the mouth and nose when expressing these reflex activities, therefore constitute an abode for bacteria. Furthermore, bacteria found in handkerchiefs could differ from one individual to another as the bacteria found could be a reflective of the environment and pathological conditions of the individual using the handkerchief. For instance, individual with upper respiratory tract infection are likely to dislodge strains of pathogenic microbes along sides with the normal flora in these regions.


1.4 Objectives of the study

  1. To enumerate the bacteria on handkerchief used by male.
  2. To identify the bacteria on handkerchief used by male.
  3. To proffer possible ways of reducing the spread of bacteria.

1.5 Significance / Justification of the study

In view of the widespread sharing of handkerchiefs and other clothing items in Nigeria, and the ability of clothing to retain microorganism, therefore Research is relevant to the isolation and characterization of microbial contaminant associated with handkerchief and to justify that users of used handkerchiefs should be public concern to all in view of likelihood of getting communicable disease.

This study will also be of importance to the educational sector, as it will improve the knowledge on the subject matter and also serve as a material guide for students, researchers and academics that are carrying out research related to this study


Chapter Five


Conclusion And Recommendations

5.1 Conclusion

From this study, it is concluded that Sample B and Sample C have the highest percentage in fungal and all the handkerchiefs have the same percentages in bacteria isolates. Sample B and Sample C Handkerchief s second hand clothes pose a greater risk to the public and microorganism isolated from second hand clothes are pathogenic to human especially among women and children.


5.2 Recommendation

The following control measures are recommended

  • Proper hygiene should be adopted.
  • Avoidance of used clothing especially for infant since their prone to infections but in the case where there is partial avoidance due to the standard of living of consumers, consumers should wash second hand clothes with strong detergents.


Enumeration And Identification Of Bacteria On Used Handkerchiefs In Males


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