A Microbial Survey of Second Hand Clothe Samples Collected From Baghdad Market
A Microbial Survey of Second Hand Clothe Samples Collected From Baghdad Market
127-136 Science
127
Nada Abdulrahman F. Al-Easawi
pathogens far away from the original contact and oils) other chemicals (such as sulfur
position with human skin, mainly when trioxide, sulphuric acid, and ethylene oxide)
moisture is present from sweat, saliva, and alkalis [8].
secretions from open pimples, wounds and Antimicrobial agents also called
spilled drinks [4]. disinfectants or sanitizers such detergents, are
In a research carried out by Nelly the either microbiocidal (kill microorganisms) or
records referred that Enterococci and microbiostatic (inhibit the growth of
Staphylococci can survive for extended microorganisms). Detergents can work in a
periods of time on materials commonly wear number of ways. Some interfere with the
by patients and health care workers and on formation of the microbial cell walls or cell
various other fabrics in the hospital membranes, usually having a microbiocidal
environment [5] while another study refers that effect. Some interfere with the synthesis of
neckties worn by doctors at an intensive care microbial proteins, nucleic acids, or essential
unit produced heavy growths of coagulase metabolites. For examples, Quaternary
negative Staphylococcus on 2/5 ties tested [3]. ammonium chlorides and alcohols may be
Studies have revealed that a variety of objects, used as antimicrobial agents [9]
such as tools, toys, and clothes, can serve as The previous study indicated that soaps can
vehicles for the transmission of Shigella sp. In remove 65 to 85% bacteria from human skin
the community, the skin infections have grown [10]. A large number of chemical compounds
significantly and the majority of these have the ability to inhibit the growth and
infections are transmitted through, shared metabolism of microorganisms or kill them.
equipment also due to contact skin-to-skin [6]. The number of chemicals in enormous,
In a study conducted by Nelly et al. (2001), probably at least 10000 with 1000 commonly
Candida, Aspergillus, Mucor, and Fusarium used in the hospitals and homes. These
which are associated with nosocomial chemicals exist as solids, liquids, and gases
infections in patients survived long periods on such as granulated detergent and soaps which
fabrics and plastics which are routinely used in are used to reduce or destroy microbes [11].
hospitals. These survival results indicate the Although fats and oils are general
potential for various fabrics and plastics to ingredients of soaps but some chemical
serve as reservoirs or vectors for fungi [5]. compounds are added to enhance the
Detergents are essential to personal and antibacterial activities of soaps [12] Transient
public health. Through their ability to loosen bacteria are deposited on the skin surface from
and remove dirt from a surface, they contribute environmental sources such as used clothes
to good personal hygiene; reduce the presence causing skin infections. Examples of such
of germs that cause infectious diseases. bacteria are Pseudomonas aeruginosa [13] and
Antimicrobial agents Kill or inhibit the growth Staphylococcus aureus [14].
of microorganisms that cause diseases. The aim of the study was to investigate
Detergents and soaps found in the home can bacteria and fungi contamination in the second
be grouped into four general Categories: hand clothes and assess the risk of
personal cleaning, cloth cleaning, dishwashing transmission of pathogenic microorganisms.
and household cleaning. Within each category Also, assess the efficiency of two kinds of
are different product types formulated with detergents to remove this bacterial
ingredients selected to perform a broad contamination.
cleaning function as well as to deliver
properties specific to that product [7]. A Materials and Methods
detergent is a chemical compound it is an Collection of samples
effective cleaning product because it contains Samples were collected from three used
one or more surfactants. The surfactants used cloth markets, including Baghdad algededa, Al
in detergents can be engineered to perform Dora, Al Binook district markets. These
well under a variety of conditions. These days Markets represent the largest second hand
detergents are made from a multiplicity of clothes in Baghdad. All samples were kept in
petrochemical chemicals (derived from fats sterile plastic bags and transferred to the
128
Journal of Al-Nahrain University Vol.20 (3), September, 2017, pp.127-136 Science
laboratory for bacterial isolation and agar plates for recovery of yeast and molds.
identification. The plates were incubated at 30˚C for 2 weeks
to 4 weeks. After incubation, the colonies on
Ready to use media the surface were counted. Mycosel agar plates
Nutrient agar and MacConkey agar were were examined for molds while Sabourands
prepared according to the instruction of dextrose agar plates were examined for yeasts.
manufacturing company. Three samples were Total microbial counts, expressed as colony
collected from each of the following forming units (CFU), were recorded for each
categories: adult's clothes, baby's clothes, and plate[16]. Appearance of isolated fungi were
fabric toys. calculated according to [17].
Isolation method Inoculum preparation
The samples which are obtained from the The samples were washed with two types of
markets using swab were inoculated to isolate detergents the Neon-trademark as soap
bacteria, for adults clothes a swab from the detergent and the granulated detergents of
entire area (under arm, neck) of the sample Al-wazeer- trademark. Each sample of (Adult
was swabbed using a sterile cotton swab clothes. baby's clothes, fabric toys) were cut
immersed in sterile water while baby clothes into two pieces, one of them was washed with
were swabbed in areas of (chest, foot). soap detergent and the other washed with
Different swaps were taken from fabric toys. granulated detergent. The Temperature of
Samples suspension was prepared by adding washing water was 500C. All samples were
the swabs to 4.5ml D.W. in a sterile tube and dried before isolation. Swabbs were taken
mixed well, then inoculated into Nutrient agar from each pairs of pieces and presented a
and MacConkey agar using ABC streaking primary procedure to isolate bacteria.
method for isolation of G-ve and G+ve
bacteria then incubated at 370C for 24 hrs[18]. Identification of bacterial isolates
Isolated bacteria were identified by using
Total bacterial count (CFU/ml) Bergey’s Manual of Determinative
Samples (10 ml) were aseptically pipetted Bacteriology, on the basis of physiological,
into a sterile Erlenmeyer flask and diluted by morphological and biochemical properties
adding 90 ml of peptone water pH 7.2, then [18].
series dilutions were done (10-2 - 10 -8). 0.1 ml
of each dilution was speared on nutrient agar Morphological characterization
and incubated at 37 ˚C for 24 hours with three Colonies developed on nutrient agar, then
replicate [15]. At the end of the incubation classified according to their shape, color, odor,
period, the colonies on the Petri plates were and their margin.
counted using digital colony counter. The
numbers of colonies were estimated as CFU/ Microscopic examination
ml according to the below equation: Microscopic examination was done for the
gram-stained slides in order to characterize
CFU /ml = gram-positive bacteria from others.
Results and Discussion
The same procedure was done before and Incidence of bacteria on Clothes Samples
after samples washing. Bacterial growth exhibited a mixture of
Isolation of Fungi colonies in the Nutrient and MaCconkey Agar,
The entire area of the sample was swabbed which represent a selective medium for
using a sterile cotton swab immersed in sterile recovering G-ve and G+ ve bacteria. This
water. The swab was used for culture in latter work included the prepartion of ferment
Sabourands Dextrose agar plates for yeasts. mannitol medium which sustains bacteria
The plates were incubated at 300C for 5 days. growth on agar containing sodium chloride
Direct imprints of 2 cm2 swatches of all the (70-100g/l), therefore, it's able to grow on
samples were cut using sterile scissors and mannitol salt agar to produce yellow colonies.
cultured directly onto the middle of Mycosel During incubation, colonies with white cream
129
Nada Abdulrahman F. Al-Easawi
Table (2)
Colonies morphology, gram stain, and biochemical tests results.
Microorganism Characteristics Staphylococcus aureus Bacillus sp. Enterobacter sp.
Colony color gold creamy creamy
Cell shape cluster short rod rod
Gram stain + - -
Catalase test + + +
Oxidase test - + -
Methyl red test - - -
Voges – Proskauer test - - +
Nitrate reduction test + + +
Citrate utilization test _ + +
Urease production test - - +
Indole production test - - -
Motility test - + +
H2S production test + - +
(+) = positive result. (-) = negative result
The results of this study showed that used temperature, such bacterial characteristics
clothes such as adults and baby's clothes as allows it to survived on fabric and clothes for a
well as fabric toys were colonized frequently long time and most of clothes and
with bacteria such as Staphylococcus aureus, undergarments frequency contact with the
Bacillus, and Enterobacteriaceae which previous owner skin, expect to colonize with
have great public health interest. In the potentially pathogenic bacteria [4]. For
environment, Staphylococci are widespread Staphylococci sp., our results come agree with
and can be found on clothing as well as those of Nelly et al[5] and Loh et al [19] who
environmental surfaces [4]. A previous study reported that S. aureus can survive for long
refered that Staphylococci can survive for periods on hospital fabrics to become
years in dust and soil particles, also it can epidemiological.
resist the dryness conditions and tolerant high
130
Journal of Al-Nahrain University Vol.20 (3), September, 2017, pp.127-136 Science
The existing Staphylococci sp. is into contact with a host, they may only need to
considered as the main cause of both sporadic be present in small numbers to infect the next
infections and epidemics of varying scope. It host [4].
causes superficial skin lesions such as boils,
pneumonia, meningitis, and urinary tract Viable count of bacterial isolates
infections; and deep-seated infections, such as Results of the current study recorded high
osteomyelitis [20]. viable counts in the N-agar plates in all cases,
In the current study greater part of tested which were reported in the order of 106 per
samples were made of polyester (fabric type) sample. Compare to before and after washing
as it mentioned in clothes label. The baby treated samples, the viable counts were found
clothes and toys made of this type of fabric. to be higher in the case of before washing
Takashima et al refer that polyester and cotton treated with detergent samples. In the case of
fabrics are colonized easily by ordinary skin Adults clothes samples, the viable counts were
bacteria. Also, they found that polyester or in the order of 103 per piece, and in the case of
acrylic fibers bound Staph. aureus and P. Baby's clothes, it was 106 per piece, while in
aeruginosa at high ratios (>80%), but P. the case of Fabric toys it was 104 per piece
aeruginosa at intermediate ratios, but cotton before washing.
fibers bound them at low ratios (<10%). Nylon The samples were washed with granulated
fibers bound Staph. aureus at low ratios, [21]. detergent showed that the viable counts of
There are two factors that control the survival bacterial isolates were the lowest compared to
of microbes in fabrics, the first is microbe soap. The details are as presented in Table (3)
characteristics and fabric properties while the and Table (4).
second is environmental factors such as
temperature, humidity, etc. If these microbes
stay viable on surfaces long enough to come
Table (3)
Effects of washing with granulated detergent on plate counts in different sample.
A1 23x102 2x101
A2 26 x102 2 x101
A3 12 x102 1 x101
Average CFU of Adults clothes 20.3 x102 1.6 x101
B1 3x104 1X102
B2 13 x104 1 X102
B3 15 x104 1 X102
Average CFU of Baby's clothes 10 x104 1 X102
F1 40x103 2 x101
F2 15 x103 1 x101
F3 30 x103 1 x101
Average CFU of Fabric toys 28.3 x103 1.3 x101
131
Nada Abdulrahman F. Al-Easawi
Table (4)
Effects of washing with soap detergent on plate counts in different samples.
A1 2x103 1x101
A2 3.7x103 1x101
A3 1.5x103 1x101
Average CFU of Adults clothes 2.4x103 1x101
B1 3x106 2x101
B2 4 x106 1 x101
B3 5 x106 1 x101
Average CFU of Baby's clothes 4x106 1.5x101
F1 2.6x104 1x101
F2 1.2 x 104 1 x 101
F3 3 x 104 2 x 101
Average CFU of Fabric toys 2.2 x 104 1.3 x 101
100000
100000
28300
10000
2030
1000
CFU/ml
C.F.U.before washing
100
100
C.F.U.after washing
16 13
10
Fig.(1): Mean bacterial counts before and after washing with granulated detergent.
132
Journal of Al-Nahrain University Vol.20 (3), September, 2017, pp.127-136 Science
Soap detergent
Results showed that bacterial counts before
washing with soap detergent were 2.4x103
C.F.U in adult's clothes. Baby's clothes and
Fabric toys had the highest contamination with
mean plate counts of 4x106 and 2.2 x 104
respectively. After washing with detergent,
adult's clothes recorded the lowest plate counts
with 1 x101 and 1.3x101 C.F.U for Fabric toys,
while baby's clothes were 1.5 x 101. Table (4)
and Fig.(2) summarize the C.F.U of the
different samples before washing and after
washing with soap detergent.
10000000 4000000
1000000
100000 22000
CFU/ml
10000 2400
1000 C.F.U.before washing
Fig.(2): Average bacterial counts before and after washing with soap detergent.
In the recent study, the clothes that washed microbes number in some clothes as compared
with soap showed a reduction in the bacterial to the lower number of microbe in healthy
plate counts Table (3) and Fig.(2) but this users used clothes[1].
reduction is less than that found in samples
washed with granulated detergent. The Diversity of Fungi Isolated from Clothes
granulated detergent may contain bactericidal Molds isolated from second-hand clothes
or bacteriostatic compounds that contribute to were identified using lacto phenol cotton blue
inhibiting microbe's growth in used clothes, mounts. These molds included Aspergillus
and the concentration of the bactericidal niger. Aspergilus flavus and Rhizopus. Table
compound in soap may be less than that of 5 shows that baby clothes recorded the highest
granulated detergent. The differences in the incedence of fungi (100%) among other used
microbes’ counts among the three samples clothes categories, while adult clothes
types and the differences in CFU before and recorded 66.6% followed by fabric toys
after washing as shown in Fig. (1, 2) could be samples which recorded 50%. A previous
attributed to different reasons such as, study by potera. 2001 referred that some fungi
Fumigation that carried out on clothes from species were isolated from the clothes used
the country of origin or some of the clothes are frequently by different individuals, such as
hanged in the open in the used clothes market Rhizopus and Aspergillus flavus these common
where the microbes can be killed by ultraviolet fungus has long been known to cause a
radiation of the sun or the frequency of usage potentially deadly threat of infections for
and laundering by the previous user, as well as hospital patients with injured or impaired
hygiene habits and their state for e.g. sick user immune systems [22].
could be considered as a source to increase
133
Nada Abdulrahman F. Al-Easawi
Table (5)
Isolated fungi sp. and their incidence percent from used clothes categories.
Number of samples incidence percent Fungi Isolate
Clothes category Isolating fungi sp.
that appeared growth % number
Aspergillus niger
Fabric toys 2 50 30
Aspergillus flavus
Aspergillus niger
Baby's clothes 3 100 49
Aspergillus flavus
Rhizopus
Adult clothes 2 66.6 223 Aspergillus niger
Aspergillus flavus
Before washing
After washing
(Soap detergent)
After washing
(Granulated detergent)
Fig.(3): Fungi species colonies before and after washing with soap and granulated detergent.
Table (6)
Spectrum of fungi isolated from used clothes categories.
C.F.U after washing with
C.F.U
Clothes category Isolating fungi sp. detergent
Before washing
Soap granulated
Aspergillus niger 20 4 1
Fabric toys
Aspergillus flavus 10 8 1
Aspergillus niger 22 2 0
Baby's clothes
Aspergillus flavus 27 7 0
Rhizopus 200 0 0
Adult clothes Aspergillus niger 3 2 0
Aspergillus flavus 2 14 0
134
Journal of Al-Nahrain University Vol.20 (3), September, 2017, pp.127-136 Science
136