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PROCESSING URINE SAMPLE. INTRODUCTION: Normally, the urine does not contain significant numbers of any microorganism.

However, if bacteria or yeast are introduced into the urinary tract, they can multiply and cause a urinary tract infection, called a UTI. Most UTIs are caused by Escherichia coli (E. coli), one of the most common human bacteria. Other frequently identified bacteria are Proteus, Klebsiella, and Staphylococcus saprophyticus. Females get UTIs more often than males. Even school-age females may have frequent UTIs. For males with a culture-proven UTI, the doctor may order further tests to rule out the presence of a kidney stone or a structural abnormality of the urinary tract that could cause the infection. The culture may be used when symptoms indicate the possibility of a urinary tract infection, such as pain and burning when urinating and frequent urge to urinate. In addition, it may be used for patients who have a catheter inserted for an extended period of time, even if they do not show obvious symptoms of an infection, since there is a risk of bacteria being introduced by the catheter. Pregnant women without any symptoms may be screened for bacteria in their urine, which could harm the baby. OBJECTIVES: 1. To process urine sample. 2. To identify pathogenic microorganism that cause s infection at urinary tract. 3. To find the suitable antibiotics to treat this type of urinary tract infections. METHODS AND MATERIALS: Wire loop and needle, Bunsen burner, gloves, glass slides, Neubauer ruling, microscope, disinfectant, normal saline, oil immersion, lens paper. Urine sample, Gram stains reagents,CLED 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) and gram positive antibiotic (Pen, Cn, FD10, E15)

PROCEDURES: FIRST DAY. 1. One drop of urine specimen was dropped into the Neubauer ruling, and counted the cells that can be observed under microscope. 2. The CLED agar was divided into two sections for colony count (a) and common culture (b).
a)colony count

b) Culture

a) The uro strip was deep inside the urine container and makes sure no excess urine at strip before tapped it slightly on the CLED agar. b) The standard streaking procedure was followed on this section. 3. The CLED agar was incubated in 37 C temperatures for 24 hours. SECOND DAY 1. The raw observation has done to the all plate and the result was 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 for gram negative mucoid bacteria 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 oxidase, catalase and coagulase test was done on gram positive bacteria. 6. 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 agars were measured using ruler, the results were recorded.

RESULTS: Gross Observation: Clear of urine sample. Lactose ferments colony growth on CLED agar, because it changes green color of CLED agar to yellow. Microscopic observations: Microscopic observations Gram stain Gram: positive Shape: cocci

Gram: negative Shape: bacilli (mucoid)

Colony Count: >25 >105 colony growth are significant. Biochemical tests for gram positive cocci :

Oxidase

Negative

Catalase

Negative

Coagulase (Slide) (tube) Negative Negative

Biochemical tests for gram negative (mucoid):

Biochemical tests Indole Motility TSI

Results Positive Motile Slant / butt / gas acid/ acid/ gas produce Non lactose ferment but glucose ferment bacteria.

MR test

Positive Change to red colour.

VP test

Negative Not cloudy

Citrase

Negative Color not change.

Sensitivity test results: a) Gram positive Antibiotics Penicilin G (Cn) Fusidic asid (FD10) Erythromycin (E15) b) Gram negative Antibiotics Ampicillin (AMP) Gentamicin (CN10) Augmentin (AMC) Cefuroxime (CXM) DISCUSSION: Urine culture result usually negative, if positive it means the infection of urinary tract. It is depend on significant growth of only one type of bacteria and the urine sample should be fresh. The mixed culture of bacteria maybe caused by contamination during collecting sample, lack of aceptic techniques during culture or the urine specimen not Diameter of clear zone (mm) 7 20 7 15 Diameter of clear zone (mm) 6 16 7 15

fresh or store in proper ways. Some urine samples have the potential to be contaminated with normal flora from the skin, care must be taken with interpreting some culture results. The presence of a single type of bacteria growing at high colony counts is considered a positive urine culture. CONCLUSION: As conclusion the infection of urinary tract is caused by E-coli. The disease are pyelonephritis, burning sensation during urinate, urgency and frequently of urination. The E-coli transmitted from GIT tract.

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