Tile Page i
Approval Page ii
Table of Content v
List of Figures
List of Table
Problem of Statement 13
and Objective of the study 15
and Method 29
Sterilization of Materials 29
3.2 Sample Size 29
3.3 Collection and Processing Sample 30
3.4 Culture Media 30
3.5 Isolation of Cultures 31
3.6 Characterization of Bacteria Isolates 32
3.7 Colonial Morphology 32
3.8 Cellular Morphology 32
3.9 Biochemical Characterization 34
3.10 Antibiotics Sensitivity Test 36
4.0 Results 39
5.0 Discussion 60
5.1 Conclusion 66
5.2 Recommendation 67
LIST OF FIGURES
Figure 1: Frequency
of Occurrence of bacterial from Wound Infection patient in University of Ilorin
Figure 2: Bacterial
Isolate from patient with diabetic ulcers in University of Ilorin Teaching
Figure 3: Bacterial
Isolated from patients with surgical wound in University of Ilorin Teaching
Figure 4: Bacterial
Isolated from patients with burns in University of Ilorin Teaching Hospital.
Figure 5: bacterial
Isolated from patient with pressure sore in University of Ilorin Teaching
LIST OF TABLE
Table 1: Morphological
Characteristics, Staining Reaction and Biochemical Test
2: Frequency of Occurrence of
bacteria from Wound Infection patient in University of Ilorin Teaching
Table 3: Bacteria pathogens causing wound infection in patients
with Diabetic ulcers.
Table 4: Bacteria pathogens causing wound infection in patients
with Surgical Wound.
Table 5: Bacteria pathogens causing wound infection in patients
Table 6: Bacteria pathogens causing wound infection in patients
with Pressure Sore.
Table 7: Antibiotic
susceptibility Patterns of Staphylococcus
aureus isolated from wound patients in UITH
Table 8: Antibiotics
susceptibility pattern of Pseudomonas
aeruginosa isolated from wound in UITH
Table 9: Antibiotic
susceptibility pattern of Staphylococcus
epidermidis in UITH.
Table 10: Antibiotic
Susceptibility pattern of Escherichia
coli isolated from wound patients in UITH.
Table 11: Antibiotic
Susceptibility pattern of Streptococcus
pyogenes isolated from wound patients in UITH.
Table 12: Antibiotic
Susceptibility pattern of Klebsiella
pneumoniae isolated from wound patients in UITH.
work investigated the antibiotic susceptibility profile of bacterial associated
with wound sepsis of patients attending University of Ilorin, Ilorin, Teaching
Hospital. Wound swabs were collected from a total number of Hundred patients
with different kinds of wound (surgical wound, burn, ulcers and pressure sores)
and cultured, of which 72 samples showed bacterial growth. Six different
species of bacteria were isolated. Staphylocococcus
aureus (47.2%) Pseudomonas aeruginosa (19.5%), Klebsiella pneumonia
(6.85%), Escherichia coli (15.3%), Staphylococcus epidemidis
(5.6%) and Streptococcus pyogenes (5.6%). The antibiotics
susceptibility test of these bacterial isolate was performed using the Kirby-
bauer dist diffusion method. Gantamycin and Sreptomycin shows high
effectiveness to all the isolate except Staphylococcus epidermis and Klebsiella with pefloxacin and ceftadine
respectively showing sensitivity to them. Resistance were shown amongst
Ampicillin, contrimoxazole and Tetracycline. This study has revealed Gentamycin
as the only antibiotic to which most bacterial isolate from infected wounds
were sensitive to. Ampicillin and penicillin were effective against Streptococcus pygenes while zinacef and ceftazidime shows high effectiveness
against Klebsiella pneumonia.
wound infection: antibiotics susceptibility profile: staphylococcus aureus:
A wound results following disruption of the
skin which can be intentional or accidental (Giacometti, 2000).Wound infections
cause a burden of disease and morbidity for both the patient and the health
services. To the patient it causes pain, discomfort, inconvenience, disability,
financial drain, and even death due to complications such as septicemia. It
causes financial strain on the health services due to the required high cost of
hospitalization and management of the patients.
number of factors contribute to wound infection; however microorganisms are the
major cause with bacteria being the most prevalent (Obuku, 2012). Early
recognition of wound infection and appropriate management is important.
Antibiotic therapy and surgical management are the cornerstone measures whereby
antibiotics offer adjuvant treatment. Wound infection can be caused by single
bacteria or multiple microorganisms. Surgical site infections are the second
most common cause of nosocomial infections after urinary tract infections
surgical site infections occur in ambulatory patients after discharge from the
beyond the hospital infection control surveillance programs (Cosgrove, 2003).
Prolonged preoperative hospital stay and exposure to diagnostic procedures has
been associated with increased rate of surgical site infection. In clean
surgical procedures, Staphylococcus aureus is the most common pathogen
while Pseudomonas aeruginosa is the most common gram negative bacilli.
number of studies indicate an increase in antibiotic resistant microorganisms
in surgical patients. Resistant bacteria causes severe infections that are
expensive to diagnose and difficult to treat. The mechanism by which resistance
develops is complex and can result in multi-drug resistant bacterial strains
due to simultaneous development of resistance to several antibiotics.
Determination of local bacterial sensitivity patterns to antibiotics is
important in providing a guide for antibiotic selection.
are factors that increase the risk of wound infection which include patient
characteristics such as; age, obesity, malnutrition, endocrine and metabolic
disorders, smoking, hypoxia, anaemia, malignancies and immunosuppressants (WHO,
2007). Other factors are the state of the wound which includes nonviable tissue
in the wound, foreign bodies, tissue ischaemia, and formation of haematomas,
long surgical procedures, and contamination during operation, poor surgical
techniques, hypothermia and prolonged pre-operative stay at the hospital.
Wound infections can be prevented by restoring
blood circulation as soon as possible relieving pain, maintaining normal body
temperature, avoiding tourniquets, performing surgical toilet and debridement
of the wound as soon as possible, administration of antibiotic prophylaxis for
deep wound and high risk infections (WHO, 2007). High risk wounds include
contaminated wounds, penetrating wounds, abdominal trauma, compound fractures,
wounds with devitalized tissue; high risk anatomical sites such as hands and
feet. Antibiotic prophylaxis should be started two hours before the surgical
of the causative microorganism is important and treatment should be initiated based
on the bacterial sensitivity patterns. Topical silver dressings have been used
to treat infected wounds however; there is no evidence for their efficacy due
to multiple microbial aetiologies. (Vermenlen, 2007). To achieve optimum
antimicrobial therapy, the biofilm load should be reduced to enhance drug
concentration at the wound site (Strup et
Bacterial wound infections are a common
finding in open injuries. Severe and poorly managed infections can lead to gas
gangrene and tetanus which may cause long-term disabilities (Bjarnsholt, 2013).
Chronic infection can cause septicemia or bone infection which can lead to
death. Sepsis associated encephalopathy increases morbidity and mortality
especially in the ICU patients (Maramattom, 2007).
Infection is an important cause of morbidity and mortality
in hospitalized burn patients in patients with burn over more than 40% of the total body surface area, 75% of
all deaths following thermal injuries are related to infections (Vindence,2005). The rate of nosocomial infections is higher
in burn patients (WHO, 2007)due to
various factors like nature of
burn injury itself, immunocompromised status of the patient (Preuitt,2008), age of
the patient, extent of injury, and depth of burn in combination with microbial
factors such as type and number of organisms, enzyme and toxin
production, colonization of the
burn wound site, systemic dissemination of the
colonizing organisms(Preuitt,2004). Moreover the larger area of tissue is
exposed for a longer time that renders
patients prone to invasive bacterial sepsis. In extensive burns when the
organisms proliferate in the eschar, and when
the density exceeds 100,000 organisms per gram of tissues, they spread to the blood
and cause a lethal bacterenia. Therapy of burn
wound infections is therefore aimed at keeping the organisms’ burden below
100,000 per gram of tissues which increases
the chances of successful skin grafting.
denatured protein of the burn eschar provides nutrition for the organisms. A vascularity
of the burned tissue places the organisms
beyond the reach
of host defense
mechanisms and systemically
administered antibiotics (Canton,2002). In addition, cross-infection results between
different burn patients due to overcrowding in burn wards. Also
thermal destruction of the
skin barrier and
concomitant depression of
local and systemic
host cellular and humeral
immune responses are pivotal
factors contributing to
infectious complication in patients with
severe burn (Maramattom,2007). Burn wound infections are largely hospital
acquired and the infecting pathogens differ from one hospital to another. The
burn wound represents a susceptible site for opportunistic colonization by
organisms of endogenous and exogenous origin; thermal injury destroys the skin
barrier that normally prevents invasion by microorganisms. This makes the burn wound
the most frequent origin of sepsis in these patients (Anguzu, 2005). Burn wound
surfaces are sterile immediately following thermal injury, these wounds
eventually become colonized with microorganisms, gram-positive bacteria that
survive the thermal insult, such as S. aureus located deep within
sweat glands and hair follicles, heavily colonize the burn wound surface within
first 48h (Oduyebo, 2008). Topical antimicrobials decrease
microbial overgrowth but
seldom prevent further
colonization with other potentially invasive bacteria and fungi. Gastrointestinal and
upper respiratory tract
and the hospital environment ( Hansbrough,2007). A susceptible
site for opportunistic colonization by organisms of endogenous and exogenous origin;
Following colonization, these organisms
start penetrating the viable
tissue depending on their
invasive capacity, local wound
factors and the degree of
the patient s
immunosuppression. If sub-eschar
tissue is invaded, disseminated infection is
likely to occur,
and the causative
infective microorganisms in
any burn facility
change with time Individual organisms are brought into
the burns ward on the wounds of new
patients. These organisms then persist in the resident flora
of the burn
treatment facility for
a variable period
of time, only
to be replaced
by newly arriving microorganisms. Introduction of new
topical agents and systemic antibiotics influence the flora of the wound
(NCCLS, 2000). The aim of the present study was to obtain information about the
type of isolates, identification and antimicrobial sensitivity of bacterial
wound infections in burn patients.
Most wound infections can be classified into
two major categories; skin and soft tissue infections although they often
overlap as a consequence of disease progression. Infections of hospital
acquired wounds are among the leading nosocomial causes of morbidity a4nd
increasing medical expense .Routine surveillance for hospital acquired wound
infection is recommended by both the centers for disease control and prevention
and surgical infection society .
The most useful classification of wound from a practical
point of view is the rank and wakefield classification (Russell et al., 2004) which classified wounds
into tidy and untidy wounds. Tidy wounds are inflected by sharp instrument and
contain no devitalized tissue. Examples are surgical incisions, cut from glass,
knife and machete. Skin wounds will usually be single and clean cut. Untidy
wounds results from crushing, tearing, avulsion, vascular injury or burns and
contain devitalized tissue skin wounds will often be multiple and irregular.
(Wikipedia, 2007) can be classified into a number of different types according
to the object that caused the wound. Types of open wound include: incision or
incised wounds, laceration, abrasions (grazes) puncture wounds and gunshot wounds,
penetration wounds and gunshot wounds. Closed wounds have fewer categories but
are just as dangerous as open wounds. The types of closed wounds include:
contusion, hematoma, crushing injuries. Bruise, contusion and Hematoma: a
closed blunt injury may result in a bruise or contusion. There is bleeding into
the tissue and visible discoloration where the amount of bleeding is sufficient
to create localize collection in the tissue, it is described as hematoma.
Puncture wounds and Bites: puncture wound is an open injury in
which foreign materials and organisms are likely to be carried deeply into the
underlying tissues. A major danger is that they may give rise to an abscess.
They are caused by an object puncturing the skin such as nail or needle.
Abrasion and Friction Burn: is a shearing injury of skin in which
the surface is rubbed off. Most are superficial and will heal by epithelization
but some may result in full thickness skin loss. It is often caused by sliding
fall unto rough surfaces.
Laceration: is the result of which a sharp object causing
irregular wounds to soft tissues which lie over hard tissue.
Traction and Avulsion: Avulsion injuries are open where there has
been a severe degree of tissue damage such injuries occur when hands or limbs
are trapped in moving machinery such as rollers.
War wounds and Gunshot injuries: are associated with different
severity of tissue damage depending on whether the injury is of low or high
Crush: are further variant of blunt injury and are often
accompanied by degloving and compartment syndrome (Wikipedia, 2007).
Ulcer: is any breach in an epithelial surface. Chronic ulcers
(Example is leg ulcer seen in diabetes mellitus patient) are wounds that fail
Pressure sore: is also known as decubitus ulcer. These are chronic
wounds following tissue necrosis from pressure. They occur over bony
prominences. They arise where there is unrelieved pressure in the soft tissue
overlying bone such that the external pressure exceeds capillary perfusion and
ischemic necrosis occurs.
Burns: are tissues from thermal (heat or cold) application or from
the absorption of physical energy or chemical contact. Types include scalds,
fat burn, flame burns, electrical burns, cold injury, friction burns, cognizing
radiation and chemical burns (Russell et
cell wall of bacteria consists of toxic lip polysaccharides called end toxins.
Many of these bacteria endotoxin are bactericidial or bacteriostatic to related
species of bacteria, but not themselves. The concept of that substances derived
from organisms may kill another (Antibiosis) is almost as old as the science of
microbiology (Bud, 2005). More than 3000 years ago ancient people stumbled over
the discovery that some India of Central America use mould to treat rashes and
the 1860s, Louis Pasteur showed that many diseases were caused by bacteria.
Later he discovered that he may be able to fight germs with other miocrobes. It
was two German doctors who were the first to make an effective medication from
microbes; Rudolf Emmerich and Oscar Low conducted their experiment in the
1890s. They proved that the germs that would cause one diseases may be cure for
another. A medication called pyocyanase was created from Bacillus pyocyaneus
and it was effective in killing the organism that caused cholera, typhoid,
diphtheria, and anthrax. It was the first antibiotic to used in hospital.
The antibiotic properties of penicillin spp. were first
described in France by Ernest Duchesne in 1897. However, his work went by
without much notice from the scientific community until Alexander Fleming’s
discovery of penicillin. Modern research on antibiotic therapy began in Germany
with the development of the narrow-spectrum antibiotic salvarasan by Paul
Ehrlich in 1909 for the first time allowing an efficient treatment of the
widespread problem of syphilis. The drug was also an effective against other
spirochaetal infected. The invention of the sulfa drug followed that of
penicillin. Sulfa drug comes from prontosil. Which is a substance used as a
dye. It has been proven that this drug could cure pneumonia, scarlet fever and
blood poisoning, (Berkow, 1999). In the 1930s other laboratories started making
other drugs in the same family. An American, Doctor Selman Waksman discovered a
drug called streptomycin. It originated from microbes found in soil and was a
cure for many intestinal diseases. Auremycin a broad spectrum antibiotic was
discovered and it was found that it could do the job of both penicillin and
streptomycin. Another laboratory discovered chloromycin. In 1949, another
laboratory came up with one of most effective antibiotics ever found,
Tetracycline. This drug could be used against many bacteria diseases (Skerrft
and Stratton, 2006).
Antibiotics can be
obtained from microorganisms, plants or could he synthetically produced, an antibiotic
is an antimicrobial substance produced by a living micro-organism and active in
high dilution (Thomas, 1988). It was defined as a metabolism product of one
organism that is detrimental or inhibitory to other organism in very small
amount by Daniel in 1992. Antibiotic is a product of the metabolism of a micro
organism that is inhibitory to other micro organisms. Antibiotics was
originated referred only to substance extracted from fungus or other organisms
but it has later come to include many synthetic and semi-synthetic drugs that
have antimicrobial or antibacterial effects (Wikipedia, 2007)
The effectiveness of antibacterial agents is influenced by
a number of factors. The concentration if it will be bacteriostatic or
bactericidal. The d\time of exposure and type of organism; the longer the
contact time, the greater would be the number of organisms killed. Organisms which clump together take a
longer time to kill than organisms that occur once. Temperature can also
influence antimicrobial activity.
Antibiotics may be categorized (Brooks et al., 2003) according to their
spectrum of activity or mode of action; Broad (affect both Gram
Positive and Gram Negative),Narrow(effect against only one group of organism)
or Limited(effective against only one or very few bacteria).They may be
classified on the basis of their mode of action(ability to interfere with
protein synthesis, structure and function of cytoplasmic membrane, DNA
synthesis, or inactive of metabolic pathways). On the basis of their chemical
structures they are classified as macrolides (erythromycin), carbohydrate
containing antibiotic (aminoglycosides), aromatic antibiotic (chloraphenicol),
quinines (tetracycliune), etc.
antibiotic is achieved by the pathogens through the following way (CDC, 2004;Prescott
et al 2003)
a) By pumping the antibiotic out of the cell after it has entered.this
pumps is known as multidrug –resistance pumps.pathogens with this pumps are Escherichia coli, Pseudomonas aeruginosa and
b) Some pathogen act by inactivating antibiotics through chemical
modes. Examples include penicillin’s enzyme action on beta lactam ring of
penicillins, phosphorylation of amino glycosides and acetylating of amino
glycoside and chloramphenicol.
Another way in which resistance to antibiotic is achieved is through the
modification of the target sites in the pathogen mutation. This is exhibited in
streptomycin resistance by Escherichia
Some bacteria lack target structures which antibiotics can attack. An example
is seen in Mycoplasma which lack bacterial cell wall hence their resistance to penicillin’s.
By preventing entrance of the antibiotic into the cells. This is seen in penicillin
resistant gram negative pathogens.
The gene responsible for antibiotic resistance
is located on the plasmid and chromosome in the pathogen (CDC, 2007).
wounds are a common cause of morbidity. Despite improvement in the practice of
medicine and attempts to provide aseptic conditions in the surgical wards, the
incidences of wound infection are increasing. Management of wound infection
remains a challenge in the surgical areas with the increasing resistance to
antimicrobials Antimicrobial resistance can lead to complications which depend
on severity that cause disability or death and increased cost of
hospitalization and management. In children, this impacts negatively on the
quality of life at a tender age. The antibiotic sensitivity patterns have not
been studied fully especially in the surgical pediatric patients at University
of Ilorin Teaching Hospital. Inappropriate antimicrobial use is associated with
increased resistance. It was therefore important to identify the causative
organisms and determine the antimicrobial sensitivity patterns to help reduce
infections and ensure appropriate use of antimicrobials.
Purpose of the Study
infection is a common problem in children, proper management with appropriate
antibiotics is therefore important to reduce morbidity that may arise. This
study aimed to determine the aetiology and antimicrobial sensitivity patterns
of bacteria that cause wound infections in the surgical paediatric patients at University
of Ilorin Teaching Hospital. Antibiotic misuse and overuse can lead to
resistance which necessitated the need for the study.
is considered one of the major health problems in the world and infection is
one of the most frequent and severe complications in patients who have
study is based on understanding that bacteria are responsible for delayed wound
healing and the so called normal flora of the skin can become pathogenic. This
study will not only identify the bacteria but also perform antibiotics
susceptibility test on each them so as to know the drug that will be effective
against a particular bacteria present in the wound.
1.4 Aims and Objective
main objective of this study was to identify the bacteria that cause wound
sepsis and their sensitivity patterns in eastern and western part of Ilorin at
the University of Ilorin Teaching Hospital.
The specific objectives were:
determine the prevalence of wound infection in the paediatric surgical
wards in University of Ilorin Teaching Hospital.
identify the bacteria that cause wound infection.
determine the antimicrobial sensitivity patterns of the isolated bacteria
find out the antibiotics used in management of wounds in the paediatric
surgical wards in University of Ilorin Teaching Hospital.
findings of the study was to help in choosing appropriate antibiotics by
considering the sensitivity patterns that were observed, hence appropriate
management of the infected wounds.
would result in a cost effective therapy for the patient and reduced financial
burden of hospitalization.
infections are caused by a number of microorganisms which include bacteria,
fungi and viruses. This study was only able to address sepsis caused by aerobic
bacteria and anaerobic bacteria. The study was not able to address the factors
that contribute to resistance patterns that were observed.