ABSTRACT
Medical
equipments (stethoscopes, thermometers and sphygmomanometer cuffs) which are
universally used as a medical device by health workers, are likely to be
contaminated by microorganisms, if they are not cleaned or disinfected may
transmit pathogens from one patient to another. A total of 30 samples were
collected from Irrua Specialist Teaching Hospital, Irrua, cultured and
incubated at 370c for 24 hours to observe for growth. Fourteen
(46.7%) of the medical equipments had bacterial contamination and sixteen (53.3%) have no significant growth. Staphylococcus aureus was the highest
contaminant with nine isolates (64.3%)
followed by Staphylococcus epidermidis
with three isolates (21.4%) and the least isolated was Staphylococcus saprophyticus with one isolate (7.1%). Different biochemical tests were done to
identify the isolates. Susceptibility to antibiotics of the isolates was also
performed. Integration of medical equipments care in daily hospital routine
enhances the control of nocosomial infections.
TABLE OF CONTENTS
Title
page i
Certification iii
Dedication
iv
Acknowledgement v
Table
of content vi
Abstract
CHAPTER ONE
1.1
Introduction 1
1.2
Objective of the study 4
CHAPTER TWO
2.1
Literature review. 5
2.2
Epidemiology 6
2.3
Cause 6
2.4
System 8
2.5
Diagnosis 9
2.6
Treatment 11
2.7
Transmission
2.8 Risk Factors 13
2.9 Prevention
16
CHAPTER THREE
3.1
Study site 19
3.2
Materials and method 19
3.3
Cleaning and
sterilization 20
3.4
Preparation for
sterilization 20
3.5
Specimen 20
3.6
Identification of
Isolates 21
3.7
Biochemical test 22
CHAPTER FOUR
Result 25
CHAPTER FIVE
5.1
Discussion 32
5.2
Conclusion and
Recommendation 35
Reference 38
Appendix
1 47
Appendix
11 50
CHAPTER ONE
1.1 INTRODUCTION
Hospital acquired infections, also known
as nocosomial infections, are infections whose development is favoured by the
hospital environment, such as one acquired by a patient during the hospital
visit, or one developing among staff. Infections are considered nosocomial when
they become clinically evident during hospitalization (at least 72 hours after
admission (Orrett et al., 1998). In
developing countries, between 5% and 10% of patients acquire one or more
infections, and 15-40% of patients admitted to critical care are thought to be
affected (Lazzari et al., 2004;
Klevens et al, 2007). In poor
settings, such as most developing countries including Nigeria, the
rate of infection can exceed 20% (Pittet, 2005; WHO, 2008). Hospital acquired
infections exact a tremendous toll, resulting in increased morbidity and
mortality, and increased health care costs (Haley, 1985). Infection
transmission in the hospital environment remains a significant hazard for
hospitalized patients and healthcare workers are potentially source of these
infections, with many pathogens transmitted by medical device such as
thermometers, sphygmomanometer cuffs and stethoscopes (Patent storm, 2004).
These medical equipments are used in hospitals by medical doctors, medical students
and other health practitioners for assessing patients’ health, have been
reported as a potential formites for transmitting infections in the hospital
environment in various parts of the world (Cohen et al., 1996; Zuliani-Maluf et
al., 2002).
There are increasing reports of the
tremendous risk of transmitting antibiotic-resistant bacteria from one patient
to another from thermometer, sphygmomanometer cuff and stethoscopes. Because
most hospital acquired infections are primarily nosocomial and not auto
infections (Hoog Kampkorstanze et al.,
1982), their acquisition in the hospital environment adds to morbidity,
mortality and economic costs (Parmar et
al., 2002). Stethoscopes, thermometers and sphygmomanometer cuffs are the
universal tools of the medical profession, are additional possible carriers, as
these come in contact with many patients, following their contact with the skin;
microorganisms can attach and establish themselves on the medical equipments
and subsequently be transferred to other patients if the medical equipment is
not disinfected before reuse (Sander, 2003; Madar et al., 2005). The transmission of infection from contaminated
medical devices can be a possible cause of hospital acquired infection
(nocosomial infection).
This
accounts for some of the difference in the knowledge, attitudes and practices
among medical professional in the use of the medical equipments (Stethoscopes,
thermometers and sphygmomanometer cuffs).
1.2
OBJECTIVES
OF THE RESEARCH
1. To
assess medical equipments (Stethoscopes, thermometers and sphygmomanometer cuffs)
handling and maintenance practices among Medical practitioners.
2. To
determine the bacterial agents that can contaminate medical equipments (Stethoscopes,
thermometers and sphygmomanometer cuffs) used by medical practitioners.
3. To
determine the antibiotic sensitivity of bacterial isolates from the medical
equipments used by
the Medcal Practitioners.
CHAPTER TWO
2.1
LITERATURE
REVIEW
The
majority of Nosocomial infections are of non minor or moderate clinical
importance but nevertheless may cause distressing morbidity, lengthen hospital
stay and increase costs. Some nosocomial infections can have catastrophic local
consequences, such as infection of a hip joint prosthesis or a prosthetic
hearth valve, although these do not necessarily result in death. Serious
nosocomial infections due to Gram negative bacilli and methicillin-resistant
staphylococci have increased greatly in incidence during the last 30 years and
the ultimate consequence of these may be fatal septicaemia (Cohen et al., 1997; Zuliani-Maluf et al., 2002).New or previously unrecognized
serious problems have occasionally occurred in recent years, such as nosocomial
infection caused by Vancomycin-resistant enterococci. Members of the hospital
staff are also sometime at risk from other disease such as tuberculosis,
hepatitis, chicken pox and human immuno deficiency virus (HIV) infections.
There are three main routes of spread of
organisms causing nosocomial infections: Self infection (autogenous infection),
Gross-infection and environmental infection. Colonization rather than infection
occurs with each of these three routes in many instance. This is particularly
relevant when considering the epidemiology of outbreak of nosocomial infection,
as the colonized patient or member of staff may cause symptomatic infection in
another patient. Examples of nosocomial infection are : Ventilator associated
pneumonia (VAP), Staphylococcus aureus
infection, Methicillin resistant Staphylococcus
aureus (MRSA) wound infections, Candida
albicans infections, Pseudomans aeruginosa
infections, Stenotrophomonos maltophilig infections, Pseudomenbraneous colitis,
Tuberculosis, Urinary tract infection, Hospital acquired Pneumonia (HAP), Gastroenteristics,
Vancomyan-resistant Enterococcus (VRE), and Legionanaire’s disease.
2.2
EPIDEMIOLOGY
Nosocomial
infections are commonly transmitted when hospital officials become complacent
and personnel do not practice correct hygiene regularly. Increased use of inpatients
treatment mean that people who are hospitalized are more ill and have more
weakened Immune systems than may have been true in the past. Moreover, some
medical procedure by pass the body’s natural protective barriers. Since medical
staffs move from patient to patient, the staffs themselves serve as a means for
spreading pathogens. Essentially the staffs and some medical equipment can
serve as vectors.
2.3
CAUSES
All
hospitalized patients are at risk of acquiring an infection from their
treatment or surgery. Some patients are at greater risk than others, especially
young children, the elderly and persons with compromised immune systems. The
National Nosocomial Infection Surveillance System Database compiled by CDC
shows that the overall infection among the children in intensive care is 6.19%,
with the primary causes being venous catheters and ventilator-associated
pneumonia (Patent storm, 2004).
The
risk factors for hospital acquired infections in children include parenteral
nutrition (tube or intravenous feeding), the use of antibiotics for more than
10 days, use of invasive devices, poor post-operative status and immune system
dysfunction. Other risk factors that increase the opportunity for hospitalized
adults and children to acquire infection are:
1. A
Prolonged hospitalization
2. Severity
of underlying illness
3. Compromised
nutritional or immune status
4. Use
of indwelling catheters
5. Failure
of health care workers to wash their hands between patients or before
procedures
6. Prevalence of antibiotic resistance
bacteria from the overuse of antibiotics. Any type of invasive procedure can
expose a patient to the possibility of infection. Some common procedures that
increase the risk of hospital acquired infections include
1. Urinary
bladder catheterization
2. Respiratory
procedures such as incubation or mechanical ventilation
3. Surgery and the dressing or drainage of
surgical wounds
4. Intravenous
procedures for delivery of medication, transfusion or nutrition
5. Gastric drainage tubes into the stomach
through the nose or mouth.
2.4
SYMPTOMS
Fever
is often the first sign of infection (Vincent, 2003). Other symptoms and signs
of infection are rapid breathing, mental confusion, low blood pressure and a
high white blood cell count. Patients with a Urinary Tract Infection (UTI) may
have pain when urinating and blood in the urine. Symptoms of pneumonia may
include difficulty breathing and cough (Cohen
et al., 1997; Zuliani-Maluf et al., 2002). A localized infection
begins with swelling, redness and tenderness on the skin or around a surgical
wound or other open wound, which can progress rapidly to the destruction of
deeper layers of muscle tissue and eventually sepsis.
2.5
DIAGNOSIS
An
infection is suspected anytime a hospitalized patient develops a fever that
cannot be explained by the underlying illness. Some patients, especially the
elderly, may not develop a fever. In these patients, the first signs of
infection may be rapid breathing or mental confusion (Saxena et al., 2005).
Diagnosis of a hospital acquired
infection is determined by:
1. Evaluation
of symptoms and signs of infection.
2. Examination
of wounds and catheter entry sites for redness, swelling or the presence of pus
or an abscess.
3. A complete physical examination and review
of underlying illness.
4. Laboratory tests, including complete blood
count (CBC) especially to look for an increase in infection fighting white
cell, urinalysis, looking out for white cells or evident of blood in urinary
tract; culture of infected area, blood, sputum, urine, or other body fluids to
find the causative organism.
5. Chest X-ray may be done when pneumonia is
suspected to look for the presence of white blood cells and other inflammatory
substances in lung tissue.
6. Review
of all procedure performed that might have led to infection.
2.6
TREATMENT
Cultures
of blood, urine, sputum, other body fluids, or tissue are important in order to
identify the bacteria, fungi, virus or other microorganism have being identified,
it will be tested again for sensitivity to a range of antibiotics so that
patients can be treated immediately and effectively with an appropriate
medicine to which the causative organism will respond to (Bryan 1986; Pitte et al., 2008). While waiting for a test
result, treatment may begin with common broad spectrum antibiotics such as
penicillin, cephalosporins, tetracyclines or erythromycins. Some types of
bacteria are becoming resistant to these standard antibiotic treatments,
especially when patients with chronic illness are given antibiotic therapy for
a long period of time when this happens, a different, more powerful, and more
specific antibiotic must be used to which the specific organism responds to
(Bryan, 1986; Pitte et al., 2000).
Two strong antibiotics that have been effective against resistant bacteria are Vancomycin
and Imipenem, although some bacteria are developing resistance to these drugs
as well. The prolonged use of antibiotics is also known to reduce the
effectiveness of the patient’s own immune system, sometimes becoming a factor
in the development of infection. Fungal infections are treated with antifungal
medications. Examples of these are amphotericin B, Nystatin, Ketoconazole, Itraconazole
and Fluconazoles.
2.7 TRANSMISSION
The
transmission of hospital acquired infections are shown below with their
description:
|
|
Route
|
Description
|
|
1
|
Contact Transmission
|
The most important and frequent mode of transmission of hospital
acquired infection (Nosocomial)
|
|
2
|
Droplet transmission
|
Occurs when droplets are generated from the source person mainly
during coughing, sneezing and talking; and during the performance of certain
procedures such as bronchoscopy. Transmission occurs when droplets containing
microorganisms from the infected person are propelled through air and
deposited on the host body.
|
|
3
|
Air Borne Transmission
|
Occurs by dissemination of either airborne droplet nuclei (small
particle residue, 5mm or smaller in size) of evaporated droplets containing
micro organism suspended in air for a long period or dust particles
containing the infectious agents. Microorganisms carried in this manner can
be dispersed widely by air currents and may be inhaled by a susceptible host
within the same room or over a long distance from the source patients,
depending on the environmental factors. Therefore, special air handling and
ventilation are required to prevent air-borne transmission. Microorganism
transmitted by air include Legionella, Mycobacterium tuberculosis
and Rubeola.
|
|
4
|
Common vehicle transmission
|
Applies to microorganisms transmitted to the host by contaminated
items such as food, water, medications, devices and equipment.
|
|
5
|
Vector borne transmission
|
Occurs when vectors such as mosquitoes, flies, rats and their vermin
transmit microorganisms.
|
Contact
transmission is divided into two subgroups which are:
i. Direct
contact transmission and
ii. Indirect
contact transmission
Rates of contact
Transmission
|
|
Routes
|
Description
|
|
1
|
Direct contact transmission
|
Involves a direct body surface contact and physical transfer of
microorganisms between a susceptible host and an infected or colonised person
turns a patient, gives a patient a bath or performs other patient care
activities that require direct personal contact. Direct contact transmission
also occurs between two patients, with one serving as the source of the
infections.
|
|
2
|
Indirect transmission
|
Involves a contact of a susceptible host with a contaminated
intermediate object, usually inanimate such as contaminated instrument or
needle. Gloves that are changed between patients. In addition, the improper
use of saline flush syringes, vials, and bags has been implicated in disease
transmission. Even when Health workers had access to gloves, disposable
needles, intravenous device and flush.
|
Factors
predisposing a patient to infection can broadly be divided in three areas:
1. People
in hospitals are usually in a poor state of health, imparing their defences
against bacteria. Advanced age or premature birth along with immunodeficiency
(due to drugs, illness or irradiation) present a general risk while other
diseases can present specific risk for instance, chronic obstructive pulmonary
disease can increase chances of respiratory tract infection (Parmar et al., 2004; Lange et al., 2000).
2. Invasive
devices, for instance incubation tubes, catheters, surgical drains and
tracheostomy tubes all bypass the body’s natural line of defence against
pathogens and provide an easy route for infection. Patients already colonized
on admission are instantly put at greater risk when they undergo an invasive
procedure (WHO, 2003).
3. A
patient’s treatment itself can leave them vulnerable to infection. Immuno-suppression
and antacid treatment undermine the body’s defence, while antimicrobial therapy
(removing competitive flora and leaving resistant organisms) and recurrent
blood transfusion have also be identified as risk factors.
2.9 PREVENTION
Hospital
acquired infections can be prevented through the various steps:
1. Adoption
of an infection control program such as the one sponsored by U. S. Centres For
Diseases Control (CDC), which includes quality of procedure known to lead
infection and monitoring program to track infection rate to see if they go up or
down.
2. Sterilization
of all reusable equipment such as thermometers, stethoscopes and sphygmomanometer
cuffs and any other device that comes to contact with the skin.
3. Strict
attention to aseptic techniques in the performance of procedures including the
use of gloves, nose masks and barriers.
4. Reduction
in the general use of antibiotics to encourage better immune response in
patients and reduce the cultivation of resistance.
5. Isolation
of patients with known contagious infection
6. Frequent
changing of dressing for wounds and use of antibacterial ointment.
7. Limitation
on the use and duration of high risk procedures such as urinary catheterization.
8. Sterilization
of medical equipment.
9. Strict
adherence to hand washing rule by Health care workers and visitors to avoid
passing infectious microorganisms to or between hospitalized patients.
10. Use
of silver alloy coated urinary catheters that destroy bacteria before they can
migrate to the bladder.
CHAPTER
THREE
MATERIALS AND METHODS
3.1
STUDY
SITE
The
research was conducted in Irrua Specialist Teaching Hospital (I.S.T.H) located
at Esan Central Local Government, Edo
State, Nigeria.
The study entails the bacterial isolates associated with medical equipments
from thermometers, stethoscopes and sphygmomanometer cuffs in various wards of
the Teaching Hospital which include the male surgical wards (M.S.W.), Female
surgical ward (F.S.W.), female medical wards (F.M.S), Ganaecology ward (G.W.)
and from Pediatric wards. The research started on December 1 and ended December
15, 2011.
3.2
MATERIALS
AND EQUIPMENT USED:
Sterile
Forceps, Petri dishes, sterile wire loops, incubator, Autoclave, Bunsen burner,
Hot Air Oven, Gas, Distilled water, conical flask, cotton wool, marking pen,
microscope, glass slides, cover slips, culture plates and Sensitivity disk.
3.3
CLEANING
AND STERILIZATION OF GLASS WARE
All
glass wares were cleaned with detergent water and rinsed with sterile distilled
water before sterilization at 1600c for 1 hour in a hot air oven.
3.4
PRPARATION
AND STERILIZATION OF MEDIA.
The
media used for this study were blood agar, chocolate agar, MacConkey agar and
Nutrient agar.
Media
were reconstituted with water according to manufacturer’s instruction (See
Appendix 1).
3.5
SPECIMEN
A
total of 30 samples were collected from medical equipment such as stethoscopes,
thermometers and sphygmomanometer cuffs from Irrua Specialist Teaching
Hospital, Irrua. A sterile swab stick moistened in sterile physiological saline
was swabbed all over the surface of each medical equipment and transferred for
analysis to the medical microbiology laboratory of Irrua specialist teaching
hospital, Irrua. All laboratory analysis were done within 1 hour of sample
collection. The swabs were directly inoculated on blood agar, chocolate agar
and MacConkey agar. The inoculated media were incubated for 24hours, 48 hours and
72 hours at 370c and then examined for bacterial growth (Cheesbrough,
2000) according to the standard protocol.
3.6
IDENTIFICATION
OF ISOLATES
GRAM STAINING: This
technique helps to group organisms into Gram positive and Gram negative. It
also shows the shapes of the organisms (rod, spiral or cocci) and the
arrangement of the organism (in chain, clusters, singly) (Cheesbrough, 2000).
Colonies of the organism were aseptically picked with a sterile wire loop and
was then emulsified in a drop of normal saline on a slide. This was allow to
air dry and then fixed with heat by passing through heat briefly (Cheesbrough,
2000).
The smear was stained first with crystal
violet solution for 30 seconds, then washed off under running water. Lugol’s
iodine solution was applied for 30 seconds. The stained smear was then
decolourised with acetone and was immediately flooded off with running water.
Then, stained using 1% neutral red for 30 seconds, after which it was flooded
off with running water. The stained slide was allowed to air dry, after which a
drop of oil immersion was put on the stained portion of the slide and the viewed
under oil immersion objective lens (100x).
All gram negative organisms were dark
red colour while the organisms that stain dark purple is Gram positive.
3.7
BIOCHEMICAL
TESTS
Biochemical
tests were carried out for identification and characterization of various
isolates.
A pure culture of each of the isolates
was used in these biochemical tests, they were obtained by sub culturing
primary plate. The following tests were carried out for identification.
CATALASE TEST
The
catalase test method used was according to Harrigan and McCane (1976). An
inoculum was taken from a pure culture of the organism which was emulsified in
normal saline on a clean grease-free slide. A
drop of hydrogen peroxide (H2O2) was added into the
suspension using a Pasteur pipette. The slide is observed carefully for gas
bubbles, if present, it indicates a positive test while the absence of gas
bubbles indicates a negative test.
COAGULASE TEST
An
inoculum was taken from pure culture of the organism with the aid of a sterile
wire loop and emulsified in normal saline on a grease-free slide, a drop of
plasma was added and the slide was gently rocked. A positive result is
represented by the coagulation of the plasma indicating the production of
coagulase. This was done to confirm the presence of Staphylococcus aureus in Gram positive cocci organisms and those
which were catalase positive. The enzyme, coagulase produced by Staphylococcus aureus converts
fibrinogen in plasma to fibrin forming agglutinates
SUGAR FERMENATION
TEST
The
test was carried out following Harrigan and McCane (1976) procedure. One
percent (1%) solution of a series of sugars were prepared in peptone water to
which neutral red indicator was added. This was then distributed equally into
bijou bottles and sterilized at 1210C for 10minutes after cooling. A
loopful of the pure colony of each test organism which were grown for three to
four hours in sterile peptone water at 370C was then inoculated into
sterile 1% sugar solution in peptone water. The set was then incubated at 370C
for 24 hours. The sugars used were Mannitol and Sucrose. The production of a
yellow to pink colour indicated acid production while the trapping of gas in an
inverted Durham’s
tube indicated gas production.
CHAPTER FOUR
RESULTS
A
total of 30 samples were taken from medical equipment (stethoscopes,
thermometers and sphygmomanometer cuffs) from Irrua specialist teaching hospital,
Irrua in various wards of the hospital - the female medical ward, male surgical
ward, Gynaecology ward, Female surgical ward and paediatric ward. Fourteen
(46.7%) of the medical equipment had bacterial contamination and sixteen (53.3%)
had no significant growth after 72 hours.
The
table below show the growth of bacteria after incubating for 24 hour, 42 hours
and 72 hours at room temperature.
Table 1:
Occurrence of microorganisms in the medical equipments
|
Hours of Incubation
|
||||
|
Ward
|
Material
|
24
|
48
|
72
|
|
F.M.W
|
Stethoscope
1 Diaphragm
|
-
|
-
|
-
|
|
|
Bell
|
-
|
-
|
-
|
|
|
Stethoscope
2 Diaphragm
|
-
|
+
|
|
|
|
Bell
|
-
|
+
|
|
|
|
Thermometer
|
-
|
+
|
|
|
|
Sphygmomanometer
cuff 1
|
-
|
+
|
|
|
|
Sphygmomanometer
cuff 2
|
+
|
|
|
|
|
Sphygmomanometer
cuff 3
|
+
|
|
|
|
M.S.W
|
Sthethoscope Diaphragm
|
-
|
-
|
-
|
|
|
Bell
|
-
|
-
|
-
|
|
|
Sphygmomanometer
cuff
|
-
|
-
|
+
|
|
G.ward
|
Thermometer
1
|
-
|
-
|
-
|
|
|
Thermometer
2
|
-
|
-
|
-
|
|
|
Stethoscope Diaphragm
|
-
|
-
|
-
|
|
|
Bell
|
-
|
-
|
-
|
|
|
Sphygmomanometer
Cuff
|
+
|
|
|
|
F.S.W
|
Stethoscope
2 Diaphragm
|
-
|
-
|
-
|
|
|
Bell
|
-
|
-
|
-
|
|
|
Stethoscope
2 Diaphragm
|
-
|
+
|
|
|
|
Bell
|
-
|
-
|
-
|
|
|
Sphygmomanometer
cuff 1
|
-
|
+
|
|
|
|
Sphygmomanometer
cuff 2
|
-
|
+
|
|
|
|
Sphygmomanometer
cuff 3
|
-
|
+
|
|
|
P.ward
|
Stethoscope
1 Diaphragm
|
-
|
-
|
-
|
|
|
Bell
|
+
|
+
|
|
|
|
Stethoscope Diaphragm
|
-
|
-
|
-
|
|
|
Bell
|
-
|
-
|
-
|
|
|
Sphygmomanometer
cuff
|
+
|
|
|
|
|
Stethoscope Diaphragm
|
-
|
-
|
-
|
|
|
Bell
|
-
|
-
|
-
|
KEY:
+
= growth
- = no growth
F.M.W
= Female Medical Ward
P. Ward = Paediatric ward
F.S.W. = Female Surgical Ward
M.S.W. = Male Surgical Ward
Table 2: Ward, material and the bacterial isolated
Table 2: Ward, material and the bacterial isolated
|
Ward
|
Material
|
Isolate
|
|
F.M.W
|
Stethoscope Diaphragm
|
Staphylococcus
Saprophyticus
|
|
|
Bell
|
Staphylococcus
aureus
|
|
|
Thermometer
|
Staphylococcus
aureus
|
|
|
Sphygmomanometer cuff 1
|
Staphylococcus
aureus
|
|
|
Sphygmomanometer cuff 2
|
Staphylococcus
epidermidis
|
|
|
Sphygmomanometer cuff 3
|
Staphylococcus
epidermidis
|
|
M.S.W.
|
Sphygmomanometer cuff
|
Staphylococcus
aureus
|
|
G.ward
|
Sphygmomanometer cuff
|
Staphylococcus
aureus
|
|
F.S.W
|
Stethoscope 2 Diaphragm
|
Staphylococcus
aureus
|
|
|
Sphygmomanometer cuff 1
|
Staphylococcus
aureus
|
|
|
Sphygmomanometer cuff 2
|
Staphylococcus
aureus
|
|
|
Sphygmomanometer cuff 3
|
Staphylococcus
aureus
|
|
P.ward
|
Stethoscope Bell
|
Staphylococcus
epidermidis
|
|
|
Sphygmomanometer cuff
|
Staphylococcus
aureus
|
KEY:
F.M.W.
= Female medical ward,
M.S.W
= Male Surgical ward,
P.
Ward = Paediatric ward
G.
Ward = Gynaecology ward
F.S.W
= Female Surgical Ward.
Table
3: Bacterial Isolates from medical
equipment with their number of isolates and their percentages
|
Bacterial isolate
|
No isolated
|
% isolated
|
|
Staphylococcus
aureus
|
9
|
64.3%
|
|
Staphylococcus
epidermidis
|
3
|
21.4%
|
|
Staphylococcus
saprophyticus
|
1
|
7.1%
|
|
Total
|
14
|
46.7%
|
From
the organisms isolated in Irrua specialist teaching hospital, Irrua, Staphylococcus aureus was the most frequent
organism with nine isolates (63.4%) followed by Staphylococcus epidermidis with three isolates (21.4%) and Staphylococcus saprophyticus was the least organism to be isolated with one
isolate (7.1%).
Table 4: Biochemical
characteristics of bacterial isolate
|
Characterization
|
S. aureus
|
S. epidermidis
|
S. saprophyticus
|
|
Gram
reaction
|
+
|
+
|
+
|
|
Catalase
|
+
|
+
|
+
|
|
Coagulase
|
+
|
-
|
-
|
|
Mannitol*
|
+
|
-
|
-
|
|
Sucrose*
|
+
|
+
|
+
|
KEY:
* = Fermentation test
+
= Positive
-
= Negative
Susceptibility pattern to some antimicrobial
agents is shown in table 5 below. Isolates of Staphylococcus aureus were more sensitive to Cefixime, Ciprofloxacin
and Gentamycin (60%), Staphylococcus
epidermidis were more sensitive to Ciprofloxacin, Gentamycin, Graxine
(100%) and Staphylococcus saprophyticus
Cefixime, were more sensitive to Ciprofloxacin, Gentamycin and Graxine (100%).
Table 5:
Susceptibility patterns of isolates.
|
No. of
Sensitive isolate (%)
|
||||
|
Antibiotics
|
Abbreviation
|
Staphylococcus
aureus
|
Staphylococcus
epidermidis
|
Staphycoccus
Saprophyticus
|
|
Erythromycin
|
E
|
4 (40%)
|
0 (0)
|
0 (0)
|
|
Cefixine
|
CEF
|
6 (60%)
|
0 (0)
|
1 (100%)
|
|
Ciprofloxacin
|
CIP
|
6 (60%)
|
3 (100%)
|
1( 100%)
|
|
Gentamycin
|
GN
|
6 (60%)
|
3 (100%)
|
1 (100%)
|
|
Augumentine
|
AU
|
0 (0)
|
1 (33.3%)
|
0( 0)
|
|
Cotrimoxazole
|
COT
|
0 (0)
|
0 (0)
|
0( 0)
|
|
Cloxaculine
|
CL
|
0 (0)
|
0 (0)
|
0( 0)
|
|
Tetracyhile
|
TET
|
0 (0)
|
0 (0)
|
0 (0)
|
|
Graxine
|
GR
|
2 (20%)
|
2 (66.7%)
|
1 (100%)
|
|
Ampicillin
|
AM
|
0 (0)
|
0 (0)
|
0(0)
|
Number
of Isolated Staphylococcus aureus =
10
Number
of Isolated Staphylococcus epidermidis
=3
Number
of Isolated Staphylococcus Saprophyticus=1
CHAPTER FIVE
DISCUSSION,
CONCLUSION & RECOMMENDATION
5.1 DISCUSSION
The
rate of contamination (46.7%) observed in this study indicates that the medical
equipment used by health care worker’s could be vectors playing a major role in
transmitting microorganisms in the hospital environment. Earlier studies have
also indicated that insufficient emphasis on consistent cleaning of medical equipment
(Stethoscopes, Thermometers and Sphygmomanometer cuffs) in the medical
curriculum are responsible for the high rate of bacterial contamination of the
health workers (Zuliani-Maluf et al.,
2002; Osorib et al., 2000). A number
of studies have demonstrated that 47-60% of medical equipment analysed were colonized
by various species of bacterial agents (Cohen et al., 1997, Smith et al.,
1996, Sander, 2003, Madar et al.,
2008).
The spectrum of organisms isolated in
this study was also isolated in a number of previous studies (Zuliani-Maluf et al., 2000l Sander, 2003; Madar et al., 2005). Of the bacteria isolated
from stethoscopes, thermometers and sphygmomanometer cuffs in this study, Staphylococcus aureus was the most
commonly isolated contaminant with 9 (64.3%), followed by Staphylococcus epidermidis 3 (2.49) and the least isolated Staphylococcus saprophitycus 1 (7.1%).
Earlier
studies showed that Staphylococcus
has developed resistance to conventional antibiotics (WHO, 2000; Sood et al., 2000) and the findings of this
research confirm this. The antibiotic
sensitivity test conducted in our study indicated that all the isolated
bacteria showed high levels of resistance to most of the antibiotics assessed
(WHO, 200; Lange et al., 2000; Uneke
and Ogbu, 2007). It is well proven that these antibiotic resistant microorganisms
are capable of initiating severe nosocomiasis in a hospital environment and
could require contact isolation and aggressive treatment to prevent their
spread (Gupta et al., 2004; Saxena et al., 2005; Breathnach et al., 1992).
This
study reinforces the dire need to revisit the medical curriculum with view to
integrating adequate cleaning of thermometers, stethoscopes and sphygmomanometer
cuffs, as a strategy of controlling nosocomial infections.
In
most health care settings, the prevention of nosocomial infections is given
serious consideration. Unfortunately however, primary attention to preventing
nosocomial infections is usually passed through high risk invasive diagnostic
tools and therapeutic health care procedures.
The importance of the simple procedures
such as hand hygiene and sterilisation of health care tools (stethoscopes,
thermometers and sphygmomanometer cuffs) have been underestimated (Sengupta et al., 2006; Madar et al., 2009). By virtue of constant contact with patients by touch
and by their medical devices, health care workers have become a potential
source of transmission of hospital acquired infections. All need to wash their
hands before and after seeing each patient. Failure to do so could facilitate
the introduction of pathogens on any device that the health worker uses
frequently, such as the thermometers, sphygmomanometer cuffs and stethoscopes.
An earlier study showed that bacterial colony counts were significantly reduced
from the stethoscopes, thermometers and sphygmomanometer cuffs after cleaning
with isopropyl alcohol, sodium hypochlorite or benzalkonium chloride (Marinella
et al., 1997).
5.2 CONCLUSION AND RECOMMENDATION
Hospital
acquired infections represent an increasing financial burden and declining
quality of health care in poor settings such as inmost developing countries
including Nigeria and these have lead to increase in morbidity and mortality of
the patient and health care workers. The need of strict prevention guidelines
is very essential (Bernard et al., 199;
Harris et al., 2000).
One possible strategy for the prevention
of hospital acquired infection outbreaks can be achieved by providing each
patient with a disposable sphygmomanometer cuffs that will remain with them
during their hospital stay and be disposed when the patients are discharged.
Likewise, providing medical devices (thermometers and stethoscopes) in each
patient room that are appropriately sanitized or disinfected between patients
can prevent the outbreak of nocosmial infections. Strict adherence to CDC
guidelines regarding hand washing, hand hygiene and use of standard precautions
also remains critical to preventing future hospital acquired infections.
Health professionals are used to
carrying stethoscopes on the neck or in briefcases and they may take it home.
The possibility of the transmission of the organisms from hospital to homes and
vice versa, with the spread of
microorganisms in the family members also needs to be explored and prevented
(Lange et al., 2000; Klevens et al., 2002).
The
need of frequent cleaning of stethoscopes, thermometers and sphygmomanometer
cuffs with alcohol will reduce the transmission rate of nocosomial infections.
Furthermore, it has been suggested that hospitals need to develop more rigorous
programs and protocols for disinfecting their medical equipment such as
thermometers, sphygmomanometer cuffs and stethoscopes (Myers et al., 1978; Jarvis, 2001).
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APPENDIX I
PREPARATION OF MEDIA:
PREPARATION OF MACCONKEY AGAR
FORMULA GRAMS/LITRE
Peptic digest of animal tissue 20.00
Lactose 10.00
Bile salts 5.00
Sodium chloride 5.00
Neutral red 0.075
Final pH (at 25ºC) 7.4±0.2
DIRECTION/PREPARATION
Suspend
550.7grams in 1000ml of distilled water. Heat to boiling, dissolve the medium
completely, sterilize by autoclaving at 151bs pressure (121ºC) for 15 minutes.
Avoid overheating. Cool to 45-50ºC and pour into sterile Petri dishes. The
surface of the medium should be dry when inoculated.
PREPARATION OF NUTRIENT AGAR
FORMULA GRAM/LITRE
Peptic digest of animal tissue 5.00
Beef extract 1.50
Yeast extract 5.00
Sodium chloride 5.00
Agar 15.00
Final pH (at 25ºC)
7.4±0.2
DIRECTION/PREPARATION
Prepare
by dissolving 28 grams
of media in 1 litre
of distilled water, heat to boiling, dissolve the medium completely. Sterilize
by autoclaving at 121ºc, 15bs pressure for 15 minutes.
PREPARATION OF BLOOD AGAR
FORMULA GRAM/LITRE
Peptic digest of animal tissue 5.00
Beef extract 1.50
Yeast extract 5.00
Sodium chloride 5.00
Agar 15.00
Final pH (at 25ºC)
7.4±0.2
Sterile Blood
DIRECTION/PREPARATION
Prepare
by dissolving 28 grams
of media in 1 litre
of distilled water, heat to boiling, dissolve the medium completely. Sterilize
by autoclaving at 121ºc, 15bs pressure for 15 minutes.
Add sterile blood and mixed thoroughly.
NUTRIENT AGAR
Composition
Ingredients Grams/litre
Meat extract 1.0
Yeast extract 2.0
Peptone 5.0
Sodium dilloride 5.0
Agar 15.0
Final PH 7.4
± 0.2
Directions/Preparation
28.0g
nutrient base powder was weighed and dissolved in 1000ml of distilled water in
a sterile conical flask. The medium was allowed to soak for 10 minutes swirled
to mix, plugged with sterile cotton wool and then sterilized by autoclaving at
1210c for 15 minutes. It was allowed to cool to 470c,
mixed well before pouring aseptically into sterile Petri-dish and allowed to
set.
APPENDIX II
REAGENTS
CATALASE REAGENT:
Hydrogen Peroxide (H2O2)
3% of solution.
It is stored in a cool
place with bottle closed and protected from light.
COMPOSITION OF GRAM STAINING REAGENTS:
PREPARATION OF CRYSTAL VIOLET SOLUTION
FORMULA GRAMS/LITRE
Crystal
violet 2.0
95% Ethyl alcohol 20.0
Ammonium oxalate monohydrate 0.2
Distilled water 40.0
DIRECTION/PREPARATION
The
crystal violet was dissolved in 20ml of 95% ethyl alcohol and 0.2grams of
ammonium oxalate monohydrate was dissolved in 20ml of distilled water. The
solution were mixed and made with another 20ml of distilled water.
LUGOL’S IODINE
SOLUTION
Lugol’s iodine 1g
Potassium ioding 2g
Distilled water 100ml
DIRECTION/PREPARATION
The
potassium iodine was first dissolve in the water, iodine crystal were then
added and the mixture shorten properly to get completely dissolved.
PREPARATION OF SAFRANIN SOLUTION
FORMULAR GRAM/LITRE
Safranin 2.5
95% ethyl alcohol 100.0
Distilled water 100.0
DIRECTION/PREPARATION
The
dye was dissolved in the alcohol, 10% of this solution was added to water
(100ml) and stored thoroughly.
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