PROCESSING CSF SAMPLE.

INTRODUCTION: Cerebrospinal fluid (CSF) analysis is a set of laboratory tests that examine a sample of the fluid surrounding the brain and spinal cord. CSF is withdrawn from the subarachnoid space through a needle by a procedure called a lumbar puncture or spinal tap. CSF analysis includes tests in clinical chemistry, hematology, immunology, and microbiology. The purpose of a CSF analysis is to diagnose medical disorders that affect the central nervous system. Some of these conditions are meningitis and encephalitis, which may be viral, bacterial, fungal, or parasitic infections, metastatic tumors, syphilis and hemorrhaging in the brain and spinal cord, OBJECTIVES: 1. To process CSF sample. 2. To identify pathogenic microorganism that cause infection at brain and spinal cord. 3. To find the suitable antibiotics to treat this type of infections. METHODS AND MATERIALS: wire loop and needle, Bunsen burner, gloves, glass slides, microscope, disinfectant, normal saline , oil immersion, lens paper. skin swab sample, Gram stains reagents, blood agar, MacConkey agar, SDA agar,Blood agar, motility agar,peptone water (for indole test),Kovac reagent ,TSI agar, Citrase, methyl red reagents, VP reagents, Muller Hilton agar, gram negative antibiotics (AMP,CN10,AMC,CXM).

PROCEDURES: FIRST DAY. 1. One drop of CSF specimen was dropped into the Neubauer ruling, and counted the cells that can be observed under microscope. 2. By following the standard streaking procedure, culture the on the blood agar, MacConkey and SDA agar.

SECOND DAY 1. The raw observation has done to the all plates and the results were recorded. 2. The standard Grams staining procedure was followed and the slides were observed under oil immersion microscope. 3. The IMViC test, motility test and sensitivity test were done by followed the standard procedures that have learn in past lab session. 4. The gram negative antibiotics such as AMP, CN10, AMC and CXM were used for sensitivity test in Muller Hilton agar . 5. The results for biochemical test and biochemical test were read after 24 hours incubate. THIRD DAY 1. Observed and recorded the results in the form, for biochemical test. 2. The diameter of clear zone on Muller Hilton agar were measured using ruler, the results were recorded.

RESULTS: Gross Observation: Cloudy appearance of CSF sample. -hemolytic and grey colony on aerobic blood agar . Lactose ferment colony, appear pink in MacConkey agar. Microscopic observations: Microscopic observations Gram: negative Shape: rod (bacilli) Gram stain

Counting cell (neubaver ruling) 2 pus cell in one cubic mm a lot of microorganism.

Biochemical tests:

Biochemical tests Results Indole Motility TSI Positive Motile Slant / butt / gas Acid / acid/ gas produce lactose ferment and glucose ferment bacteria. MR test Positive Change to red colour. VP test Negative Not cloudy Citrase Negative colour not change from green.

Sensitivity test results: Antibiotics Ampicillin (AMP) Gentamicin (CN10) Augmentin (AMC) Cefuroxime (CXM) DISCUSSION: A lumbar puncture to withdraw a small amount of CSF for analysis may lead to serious complications. Lumbar punctures should be performed only with extreme caution, and only if the benefits are thought to outweigh the risks. In people who have bleeding disorders, lumbar puncture can cause hemorrhage that can compress the spinal cord. If there is increased spinal column pressure, as may occur with a brain tumor and other conditions, removal of CSF can cause the brain to herniated, compressing the brain stem and other vital structures and leading to irreversible brain damage or death. Bacteria introduced during the puncture may cause meningitis. For this reason, aseptic technique must be followed strictly, and a lumbar puncture should never be performed at the site of a localized skin lesion. Specimens should be handled with caution to avoid contamination with skin flora. They should be refrigerated if analysis cannot be performed immediately. Diameter of clear zone (mm) 0 (resistant) 8 12 10

CONCLUSION: