ABSTRACT
The aim of this study is the microbiological study of some selected cosmetics products used by their consumers. A Total Eight (8) samples of cosmetics products Powder, Lipstick, Body lotion ,Nail polish, Tiro, Enchanter powder, Body butter cream and Face scrub were collected from female students in Michael Okpara University of Agriculture, Umudike and was transported to the laboratory for microbiological analysis. A Tenfold serial dilution of each of the samples was prepared using sterile distilled water. 0.1 mL aliquots of the serially-diluted samples (10-4 and 10-5) were spread-plated on the surface of nutrient agar (NA), MacConkey agar and Blood agar contained in petri dishes to isolate the bacteria. The plates were incubated at 37˚C for 24-48hrs in an inverted position. The colonies that developed were counted, subcultured and stored on sterile nutrient agar slants for characterization and identification. The same procedure was used in the isolation of the fungi except that Saboraud dextrose agar (SDA) was used as the growth medium and the plates incubated at 370C. After microbial analysis, Six different bacterial isolates from cosmetic products were identified by morphological characteristics the major bacterial isolates to belong to Bacillus specie, E. coli, Staphylococcus aureus, Pseudomonas aeruginosa, Proteus sp and Micrococcus sp respectively while Three fungal isolates from cosmetic products which were identified by their morphological characteristics and revealed the fungal isolates to belong to Aspergillus, Aspergillus flavus, and Rhodotorula sp. The microbial evaluation of used cosmetic products exhibited high bacterial count ranging between 8.2 x 104 to 4.7 x 104 while total coliform plate count ranged from 7.8 x 104 to 4.6 x 104 and total fungal plate count has 5.0 x 104 to 2.0 x 104 respectively while the unsued cosmestic products showed that Total Heterotrophic count ranged from 8.8 x104 to 3.6 x104, while total coliform plate count ranged from 9.9 x104 and Total plate count 4.0x 104 to 3.4 x104.The percentage occurrence of bacterial isolates from different cosmetic products Staphylococcus aureus (28%) was predominant among the samples used in this study followed by E. coli (20%), Micrococcus sp (20%), Bacillus sp (16%), Proteus (12%) while Pseudomonas aeruginosa (4%) was least predominant while the percentage occurrence of fungal isolates from different cosmetic products. Aspergillus fumigatus 46%, was predominant followed by Aspergillus flavus (30.8%) while Rhodotula sp (23%) was least predominant.
TABLE OF CONTENTS
Title
Page i
Certification
ii
Dedication
iii
Acknowledgements
iv
Table
of Contents v
Lists
of Tables vii
Abstract
viii
1.0 CHAPTER ONE
1.1 Introduction 1
1.2 Types of Cosmetics 1
1.3
Where They Can Be Bought 5
1.4
Uses of Cosmetics 5
1.5
Group of Individuals That Use Cosmetics 5
1.6 Method Used In
Detecting the Microbial Load in Cosmetics 5
1.7 Advantages and
Disadvantages of Cosmetics 8
1.7.1 Disadvantages 8
1.8 Aims and Objectives 8
2.0
CHAPTER TWO
2.1
Literature Review 9
2.2
Microorganisms and Cosmetics 9
2.3 Cosmetic Preservatives 16
3.0
CHAPTER THREE
3.1 Materials and Methods 18
3.2 Study Area 18
3.3 Sample Collection 18
3.4 Media Used 18
3.5 Sterilization 19
3.6
Isolation of Microorganisms 19
3.7
Characterization and Identification of the Bacterial Isolates 19
3.8 Gram
Staining 19
3.9 Biochemical Cultural Characteristics 20
3.9.1Catalase
Test 20
3.9.2
Coagulase Test 20
3.9.3
Citrate Test 20
3.9.4
Motility Test 21
3.9.5
Indole Test 21
3.9.6
Oxidase Test 21
3.9.7 Sugar Fermentation 22
3.10
Characterization and Identification of the Fungal Isolates 22
3.10.1
Lactophenol Cotton Blue Staining 22
3.10.2
Slide Culture Test 23
3.11
Statistical Analysis 23
4.0 CHAPTER FOUR
4.1 Results 24
5.0 CHAPTER FIVE
5.1 Discussion,
Conclusion and Recommendation 30
5.2 Discussion 30
5.3 Conclusion 32
5.4 Recommendation 33
References 34
Appendix I
Appendix II
LIST OF TABLES
Table Title Page
1: Total viable microbial count of
microorganisms from different cosmetic products 25
2: Identification and Characterization of
Isolates from different cosmetic products 26
3: Morphological Identification and Characterization
of Fungal Isolates from
different cosmetic products 27
4: Percentage Occurrence of Microbial
Isolates from different cosmetic products 28
5: Percentage occurrence of fungal isolates from different
cosmetic products 29
CHAPTER
ONE
1.1 Introduction
Cosmetic product means any substance or
preparation intended for placing in contact with the various external parts of
the human body/ or with the teeth and the mucous membranes of the oral cavity
(Blanchin et al., 2014).The microbial contamination of personal care
products may already occur in the course of production, through raw materials,
ingredients, and during handling, or through repeated use by the consumer.
Cosmetics are substances or products used to enhance or alter the appearance of the face or fragrance and texture of
the body. Many cosmetics are designed for use of applying to the face, hair,
and body. They are generally mixtures of chemical compounds; some being derived from
natural sources (such as coconut oil), and some being synthetics
or artificial. Common cosmetics include lipstick, mascara, eye shadow, foundation, skin cleansers and body lotions, shampoo and conditioner, hairstyling products (gel, hair spray, etc.), perfume and cologne. Cosmetics applied to the
face to enhance its appearance are often called make-up or makeup.
In
the United States., the Food and Drug
Administration (FDA), which regulates cosmetics, defines cosmetics as
"intended to be applied to the human body for cleansing, beautifying,
promoting attractiveness, or altering the appearance without affecting the body's
structure or functions". This broad definition includes any material
intended for use as a component of a cosmetic product. The
FDA specifically excludes soap from this
category.
1.2
TYPES OF COSMETICS
Cosmetics are intended
to be applied externally. They include, but are not limited to, products that
can be applied to the face: skin-care creams, lipsticks, eye and facial makeup, towelettes, and colored contact lenses; to the body: deodorants, lotions, powders, perfumes, baby products, bath
oils, bubble baths, bath salts, and body butters; to the hands/nails:
fingernail and toe nail polish, and hand sanitizer; to the hair: permanent
chemicals, haircolors, hair sprays, and gels.
A subset of cosmetics is
called "makeup", refers primarily to products containing color
pigments that are intended to alter the user's appearance. Manufacturers may
distinguish between "decorative" and "care" cosmetics
therefore cosmetics that are meant to be used on the face and eye area are
usually applied with a brush, a makeup sponge, or the fingertips.
Most
cosmetics are distinguished by the area of the body intended for application.
- Primer comes in formulas to suit
individual skin conditions. Most are meant to reduce the appearance of
pore size, prolong the wear of makeup, and allow for a smoother
application of makeup. Primers are applied before foundation or eyeshadows
depending on where the primer is to be applied.
- Lipstick, lip gloss, lip liner, lip plumper, lip balm, lip stain, lip conditioner,
lip primer, lip boosters, and lip butters: Lipsticks are intended to add
color and texture to the lips and often come in a wide range of colors, as
well as finishes such as matte, satin, and lustre. Lip stains have a water
or gel base and may contain alcohol to help the product stay on leaving a
matte look. They temporarily saturate the lips with a dye. Usually
designed to be waterproof, the product may come with an applicator brush, rollerball, or could be
applied with a finger. Lip glosses are intended to add shine to the lips
and may add a tint of color, as well as being scented or flavored for a
pop of fun. Lip balms are most often used to moisturize, tint, and protect
the lips. Some brands contain sunscreen.
- Concealer covers imperfections of the
skin. Concealer is often used for any extra coverage needed to cover
acne/pimple blemishes, undereye circles, and other imperfections.
Concealer is often thicker and more solid than foundation, and provides
longer lasting, more detailed coverage as well as creating a fresh clean
base for all the rest of the makeup.
- Foundation is used to smooth out the face by covering spots, acne, blemishes, or uneven skin
tone. These are sold in a liquid, cream, or powder, or more recently in a
mousse. Foundation provides sheer, matte, dewy or full coverage.[2] Foundation primer is applied
before foundation to fill out pores, create a dewy look or create a
smoother finish. They usually come in cream formulas to be applied before
foundation as a base.
- Face
powder sets the foundation and under eye concealer, giving it
a matte finish while also concealing
small flaws or blemishes. It can also be used to bake the foundation, so that it
stays on longer. Tinted face powders may be worn alone as a light
foundation so that the full face does not look as caked-up as it could.
- Rouge, blush, or
blusher is cheek coloring to bring out the color in the cheeks and make the cheekbones appear more defined. Rouge
comes in powder, cream, and liquid forms. Different blush colors are used
to compliment different skin tones.
- Contour
powders and creams are used to define the face. They can give the illusion of a
slimmer face or to modify a face shape in other desired ways. Usually a
few shades darker than the skin tone and matte in finish, contour products
create the illusion of depth. A darker-toned foundation/concealer can be
used instead of contour products for the same purpose.
- Highlight, used to draw attention to the
high points of the face as well as to add glow; comes in liquid, cream,
and powder forms. It often contains a substance to provide shimmer.
Alternatively, a lighter-toned foundation/concealer can be used.
- Bronzer gives skin a bit of color by
adding a golden or bronze glow and highlighting the cheekbones, as well as
being used for contouring. Bronzer is considered to be more of a natural
look and can be used for an everyday wear. Bronzer enhances the color of
the face while adding more of a shimmery look. It comes in either matte, semi
matte/satin, or shimmer finishes.
- Mascara is used to darken, lengthen,
thicken, or draw attention to the eyelashes. It is available in various
colors. Some mascaras include glitter flecks. There are many
formulas, including waterproof versions for those prone to allergies or sudden tears. It is often
used after an eyelash curler and mascara primer. Many
mascaras have components to help lashes appear longer and thicker.
A wide range of preservatives has been
developed to combat the contamination from the repeated use by the consumer
(NakiSiviri et al., 2006). Nowadays, maintaining a balance between
protection against microbial contamination and limiting the health risks of
preservatives has constituted the art of preservation (Wu et al., 2010; Taher,
2011).
1.3
WHERE THEY CAN BE BOUGHT
Cosmetic products can be bought at
pharmaceutical shops, Supermarkets or Cosmetic shops.
1.4
USES OF COSMETICS
Cosmetics are substances or products used
to enhance or alter the appearance of the face or fragrance and texture of the
body. Many cosmetics are designed for use of applying to the face, hair and
body.
1.5
GROUP OF INDIVIDUALS THAT USE COSMETICS
Cosmetics are more commonly used by
females than males; however there are some useful cosmetics applications that
men can use in today’s world.
1.6
METHOD USED IN DETECTING THE MICROBIAL LOAD IN COSMETICS
Methods to detect microbial contamination
in cosmetics and their raw materials are usually based on traditional plate
counts. However, little is known about the metabolic state of microorganisms
residing in cosmetic products or in specific areas of a manufacturing plant.
Viable microorganism are often metabolically injured as a result of adverse
physical or chemical conditions (high processing temperature, cleaning,
sanitization agents and preservatives). As a result, these microorganisms are
in a viable but non-cultural state and thus cannot multiply in a nutritive agar
medium.The recovery of stressed microorganisms is a challenge for cosmetic
microbiologists, since appropriate diluents, preservatives, neutralizing
agents, culture media, etc. are needed.The validation of microbiological
detection methods is therefore an indispensable prerequisite for the detection
of microorganisms. A variety of new methods, such as Bioluminescence, impedance
and cytometry, which are based on the metabolic state of microorganisms, are
the most reliable for detecting stressed cells. These “fast” methods allow the
detection of microbial contamination ,both in the finished products and in raw
materials with in 24 hours .Fast methods are of great industrial
importance,since they facilitate the rapid release of products into the
market.However,despite the advantages offered by fast methods,they are not yet
able to detect specific microorganisms,including pathogens.Thus,classical
microbiological approaches remain indispensable for the isolation and
identification of microorganisms (Leranoz, 2014).
Historically,cosmetic term includes the
products whose purpose was to enhance the appearance or modified the odour of
the human body.However this concept has evolved due to the pressure to
continually adapt cosmetics to changing market demands and to fulfill the
consumers need and expectations.This rapid evolution forced the authorities of
different countries,in the middle of the 20th century to regulate cosmetic
products in order to ensure consumer safety.
However, with the continuous increased in
the variety of raw materials and cosmetic products,it is frequently necessary
to rely on pharmaceutical, food regulations and on pharmacopoeias due to the
lack of official cosmetic guidelines.While regulation regarding microbiological
content in cosmetic products do not existfoods are classified according to
their nature and health regulations including microbiological limits are
defined.Although some recommendations have been published by governments and
cosmetics associations,the only requirement of cosmetics is that they “must not
cause damage to human health when applied under normal or reasonably
foreseeable conditions of use”.
Differences between cosmetics and
food/pharmaceuticals are often a matter of disagreement.Health authorities
would like cosmetic companies to achieve the “gold standards” imposed by
pharmaceutical regulations and recommendations on areas such as manufacturing,
filling and testing. While the general intention is correct,cosmetics are not
meant to be ingested or injected in the human body nor are they to be used for
therapeutic purpose.
Microbiologists working in the fields of
cosmetics are frequently required to design preservative systems that provide
good protection of cosmetic products against microbial
contamination.However,scientific information on this issue is scarce,since most
biocide studies deal with antibiotics for human treatment. Microbiologists must
therefore work within a narrow range of preservative concentrations in order to
achieve effectiveness against microorganisms while avoiding toxicity for
consumers.For this reason,regulations in the EU and in other countries have
specified preservatives allowed,their maximum concentrations and other
directions specifically related to the kind of cosmetic products.
Over 150 mascaras representing eight
popular brands were examined for their susceptibility to microbial
contamination during their use by study group members.Early in the studies,
brands without preservatives supported reproducing populations of microorganisms,including
potential eye pathogens. Microbes associated with the facial skin and the
fingers of the study group users were typically isolated from mascaras after
use .Four patients with Staphylococcal blepharitis and cosmetic heavily laden
with Staphylococcus epidermidis showed marked clinical improvement when
they stopped using the contaminated cosmetics .The application of used eye area
makeup prior to and following ocular surgery should be avoided (Ergun, 2007).
Antimicrobial of antibiotics was checked on bacteria isolated. Antibiotics are
among the most frequently prescribed medication in modern medicine.Some
antibiotics are bactericidal,means that they work by killing bacteria.Other
antibiotics are bacteriostatic,means that they work by stopping bacteria
multiplication.The selective action of antibiotics upon bacteria and other
microorganism is known as the Antibiotic Spectrum. Some are active against
certain bacteria and not upon others whereas some are active against fungi and
viruses (Davies, 1994). Mechanism of
action is inhibiting protein synthesis on the ribosomal level (Smadel, 2003).
1.7 ADVANTAGES AND DISADVANTAGES OF COSMETICS
1. Cosmetics are regarded
as a means of enhancing ones complexion and beautifying the skin.
2. Skin creams and lotions can be used to
deepcleanse the skin.
3. If pores in the skin
are blocked, it may lead to infection and acne formation. Various cosmetic
cleansing products thoroughly cleanse the skin and remove dirt and
microorganisms from the skin.
1.7.1 Disadvantages
Cosmetics are not valid
line of acne control, but they can camouflage an acute outbreak. One must
remember, however, that any kind of topically applied oil based compound will
aggravate acne. Cosmetics are not good for proper function of the human body. Even
natural cosmetics may contain products which are harmful.
1.8
AIMS AND OBJECTIVES
The aim of this study to evaluate the
microbial load of cosmetic products.
The objectives are:
1. To isolate the bacterial load of
used and unused cosmetic products by various consumers.
2. To identify the fungal load of
used and unused cosmetic products by various consumers.
3. To determine the percentage
occurrence of the different microorganisms that survive in cosmetics.
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