Saturday, 14 January 2017

Isolation, Characterization And Susceptibility Study Of Bacteria Isolated From Burn Wounds

The study is aimed at determining the susceptibility pattern of bacteria associated with burn wound infection. Fifty (50) swabs samples were collected from patient with burn wound infection and cultured on Macconkey and Blood agar. Seven (7) bacteria speciesStaphylococcusaureus,Escherichiacoli, Pseudomonasaerugnosa,Streptococcusspecies, Staphylococcusepiderrmidis, Klebsiella species and Micrococcus species were isolated. The percentage occurrence of the isolates werestaphylococcusaureus(18%),Escherichiacoli(14%),Pseudomonasaerugnosa(12%),Streptococcusspecies(12%), Staphylococcusepiderrmidis(10%),Klebsiella species (12%) and Micrococcus species (10%).The percentage occurance of staphylococcus aureus is higher than Escherichiacoli, Pseudomonasaerugnosa,Streptococcusspecies, Staphylococcusepiderrmidis,Klebsiella species and Micrococcus species
The antibacterial activities of the isolates were tested against some antibiotics, which include Gentamycin, Erythromycin, Pefloxacin, Amoxicillin, Ampiclox, Zinnacef, Roceptin, Streptomycin, Septrin, Chloramphenicol, Sporfloxacin, Augmentin, Tarivid and Ciprofloxacin. The result obtained showed that ciprofloxacin, Septrin,Zinnacef,Erythromycin and Streptomycin are susceptible to the isolates while Gentamycin, Pefloxacin, Amoxicillin, Ampiclox, Roceptin, Chloramphenicol, Sporfloxacin, Augmentin and Tarivid are resistant. The increase in drug resistance is a major obstacle in the treatment of bacterial infection. So the knowledge of antibiotic susceptibility pattern is essential to give proper antibiotic therapy and to avoid unnecessary medication with non- effective drug which may increase resistance.   

Tittle page      
Table of content 

2.1Anatomy of the skin      
2.3 Pathophysiology      
2.3. 1 Zone of coagulation 
2.3.2  Zone of Stasis         
2.3.3  Zone of Hyperemia  
2.3.4  Types of Burn Wound
2.4 Pathogenesis of Burn Wound Infection       
2.5 Types OF Burn Infections      
2.5.1 Burn Wound Impetigo            
2.5.2 Open Burn associated with Surgical Wound Infection  
2.5.3  Burn Wound Cellulites
2.5.4 Invassive Infection in Unexcited Burn Wounds   
2.6 Microorganism Associated with Burn Wound Infections   



3.2 Media Preparations    

3.2.1Preparation of Mueller Hinton Agar

3.2.2 Preparation of Macconkey Agar  

3.2.3 Preparation of Blood Agar     

3.2.4 Preparation of Chocolate Agar  

3.2.5Preparation of Brain Heart Infusion Broth   

3.3 Collection of Samples

3.3.1Burn Wound Swab Collection 

3.4 Isolation of Bacteria from Burn Wound

3.5 Biochemical Identification of Each Bacteria

3.5.1Gram Staining 

3.5.2 Catalase Test  
3.5.3 Coagulase Test  
3.5.4 Motilty Test 
3.5.5 Indole Test   
3.5.6 Citrate Utilization Test

3.5.7 Oxidase

3.5.8Triple Sugar Iron Agar (TSI) Test 
3.5.9 Methyl Red Voges – Proskauer Test (MR – VP) 

3.6 Antimicrobial Susceptibility Testing  

3.6.1Preparation of McFarland Standard  

3.6.2Antibiotic Susceptibility Test  



4.1 Isolation and Identification of BacteriaAssociated with Burn Wound Infections
4.2 Percentage Rate of Occurrence of the Bacteria Isolate
4.3 Susceptibility pattern of the Isolate to Some Antibiotics


5.1 Discussion

5.2 Conclusion

5.3 Recommendations

            CHAPTER ONE

Burn wound infection is an important cause of morbidity and mortality in hospitalized patients. The rate of nosocomial infections is higher in burn patients due to various factors like nature of burn injury itself, immunocomprimised status of the patient, age, extent of injury, and depth of burn in combination with microbial factors such as type and number of organisms, enzymes and toxin production, colonization of burn wound site, systemic dissemination of the colonizing organisms (Mehedi et al., 2013).

The skin provides primary protection against infection by acting as a physical barrier. However, when this barrier is damaged, pathogens have a direct route to penetrate the body, possibly resulting to infection, due to the importance of skin as a barrier to microbial host penetration, it is surprising that the risk of  burn wound infection and endemic infection correlate with the size of wound injury because 75% mobility is associated with burn wound infections (Alghalibi et al., 2011).

Burn wounds are damage to the skin and are caused by a variety of non-mechanical sources including chemicals, electricity, heat, sunlight or nuclear radiation (Alghalibi et al., 2011).  Burn injury destroys the physical barriers that normally prevent the penetration of microorganisms. They denature protein of the burn eschar provide nutrition for the microorganisms. A vascularity of the burned tissue places the organisms beyond the reach of host defense mechanisms (Mehedi et al., 2013).

Burn wounds have three separate zones of concern. The zone of coagulation is located where the skin comes in contact with the burn source, at the center of the wound. It is made up of dead, leathery tissue that forms the burn eschar (scab). The zone of stasis surround the zone of coagulation; tissue in this zone is alive but at high risk of infection and necrosis (tissue death) due to decreased perfusion, a result of poor circulation to the area. Lastly, the zone of hyperemia surrounds the zone of stasis and contains healthy skin. vasodilation is common in this area as a result of the injury (Church et al., 2006).

Burn wound infection are suitable site for multiplication of bacteria and are most lasting richer sources of infection than surgical wounds, mainly because of the larger area involved and longer period of patient stay in the hospital(Alghalibi et al., 2011).

Burns can be classified into three primary types of burns:

Ø  First degree burns: red, non-blistered skin

Ø  Second degree burns: blisters and some thickening of the skin

Ø  Third degree burns: widespread thickness with a white, leathery appearance.

There are also fourth degree burns. This type of burn includes all of the symptoms of a third degree burn and also extends beyond the skin into tendons and bones (Church et al., 2006).

The area of a burn is often determined using the “Rule of Nines,” which divides the body up into sections that correspond with approximately 9% of the body’s surface area (Church et al., 2006).

Burn associated infections are highly drug resistant pathogens with limited treatment options.
The most commonly isolated organisms from burn patients are Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Klebsiella pneumonia, Staphylococcus epidermis’s, Acinetobacter baumannii and Anterobacter cloacae(Alghalibi et al., 2011).

A high proportion of Enterobacteriaceae strains produced extended spectrum beta lactamase enzyme and Staphylococcus aureus isolates were uniformly methicillin resistant. For Gram  negative bacteria, the most reliably active antibiotics were imipenen and amikacin (Mehedi et al., 2013).

                  Aim Of The Study

The aim of this project is to determine the antibiotic susceptibility pattern of some antibiotics against different bacteria isolated from burn wound infections.

           Objectives Of The Study
1. To isolates bacteria associated with burn woundsinfection.

2. To characterize the bacteria isolates associated with burn wound infections.

3. To determine the susceptibility pattern of the bacteria isolated from burn wound infections to some antibiotics

             Statement of the Research Problem

               Burns wound infection is a major cause of morbidity and mortality. About 75% of motility associated with burn injuries is associated with sepsis rather than osmotic shock or hypovolemia. About 50% to 60% of deaths in burn patient is caused by infection in spite of intensive therapy with antibiotics both topical and intravenous (shahzad et al., 2012).
Burn wound infection is problematic because it delays healing, encourages scarring and may result in bacteremia, sepsis or multi – organ dysfunction syndrome whereby organs from several systems are unable to maintain homeostasis on their own, requiring immediate medical attention (Church et al., 2006).

            Justification Of The Study

This study will help to understand the microorganisms associated with burn wound infections. Overcrowding in burn unit is an important cause of cross infection which necessitates a regular monitoring of bacteria species and their antibiotic susceptibility (Magnetet al., 2013).The study will also help to assess the burden of infections and consequently formulation of antibiotic policy (Ahmed et al., 2006).

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