Exp. No.4(c) Processing of clinical samples: Pus

 

Pus Sample

Introduction

Wound is an abnormal break in the skin or other tissue, which allows blood to escape. Wounds are two types, they are open wound and closed wound. Open wound allows blood to escape from the body. Here the skin is broken. Open wounds are contaminated by germs, which enter from air, fingers and other parts of the body. Any wound, which has not begun to heal properly after 48 hours it, may be infected. Infection may further spread and cause dangerous illness to human beings. Staphylococcus aureus mostly isolated from skin wounds, Pseudomonas aeruginosa is associated with infected burns and with hospital acquired infections. Escherichia coli, Proteus species associated with abdominal abscess. Clostridium perfringens is found mainly in deep wounds. Possible Pathogens are Staphylococcus aureus, Pseudomonas aeruginosa, Clostridium spp., Streptococcus pyogenes. sp., Escherichia coli, Proteus sp., Bacteroides, Klebsiella sp.

Aim

To isolate the pathogenic bacteria from the wound specimens.

Materials and Methods

Sterile leak proof container, Sterile cotton swab, Aimers transport medium, Blood agar, Neomycin blood agar, Robertson cooked meat medium, Mac Conkey agar. Pus from wound is collected at the time of the abscess is incised and drained or after it ruptured naturally. Collect a sample of the pus on a sterile cotton swab and insert it in a leak proof container of Amies transport medium.

Procedure

·         Inoculated two blood agar, one neomycin blood agar, one MacConkey agar and one cooked meat medium tube using pus sample.  Incubated One blood agar and Mac Conkey agar aerobically at 35-37°C overnight. Incubated Neomycin blood agar and 2nd blood agar plate anaerobically at 35-37°C for 48 hours. Incubated Cooked meat medium at 35-37°C for 72 hours. Observed for Colony morphology on different medium and record the results. Subjected the isolated colonies to biochemical test for species identification.

 Result

Organisms isolated from wound are Staphylococcus aureus, Streptococcus pyogenes and Pseudomonas spp.