Answers: Wide Mouthed Container

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Answers

What is your diagnosis?


It could be a case of urinary tract infection. Some of the differential diagnoses include
urethritis, PID, endometriosis, vaginitis and renal calculi.

Which are the bacteria that can cause urinary tract infection?
Common uropathogens of community acquire UTI include E.coli, Klebsiella
pneumoniae, Proteus sps, and Enterobacter sps. In hospitalized patient, who have a
urinary catheter,  Pseudomonas sps, and Enterococci are common pathogens.
Staphylococcus saprophyticus is known to cause UTI in sexually active young
women.

What is the common source of UTI?


In community acquired UTI, the uropathogens frequently are one's own enteric flora.
UTI is more common in women than men due to proximity of anus to vagina and
shorter urethra.

How are urinary tract infections classified?


UTI may be community acquired or hospital acquired, lower or upper, ascending  or
descending, uncomplicated  or complicated.

How is the sample collected for laboratory diagnosis?


An early morning, freshly voided, clean-catch, mid-stream urine should be collected
in a sterile, wide mouthed container after proper anogenital toilette. The external
genitilia must be cleansed with mild antiseptic or soap before sample collection to
avoid contamination of the urine by normal flora present in this region. In men, the
prepuce is retracted and in women, the labia is spread apart and then the middle
portion of the urine is collected in the container. The sample must be labeled and sent
to the laboratory without delay.

Which are the other techniques to collect urine specimen?


In infants urine flow may be stimulated by tapping just above the pubis with two
fingers at one hour after a feed. One tap per second is given for one minute and after
an interval of one minute tapping is continued. Under certain conditions, suprapubic
aspiration of urine directly from the bladder may be performed. Since this is an
invasive technique, it must be performed only when absolutely necessary.
Catheterization only for the purpose of collecting urine should be avoided as it may
induce infection. In situations where the patient is already catheterized, the urine must
not be collected from the bag, instead, it should be aspirated from catheter tube using
needle and syringe.
How long can the urine be held before testing?
Ideally, urine must be processed as soon as possible since urine supports growth of
bacteria. In case of delay of 1-2 hours the sample may be refrigerated or treated with
boric acid at an concentration of 1.8%. Another way of preserving the sample in case
of delay is by collecting urine in sterile vacutainer tubes containing boric acid-sodium
formate transport medium. Samples that have been processed after a delay of five
hours or more do not give reliable results.

Which investigations are performed on urine sample?


Urine wet mount and culture is commonly performed on urine specimen. Wet mount
examination is performed to look for pus cells, RBCs and casts. A loopful of well
mixed urine placed on the glass slide (without spreading) can be stained by Gram
stain and observed. Presence of single bacterium per oil immersion field in such a
smear indicates significant bacteriuria. Screening test such as nitrate
reduction, dipstick, tetrazolium reduction etc are not specific and are not routinely
done. Leukocyte esterase dip test is helpful in detecting pyuria. Qualitative culture
technique such as Miles and Misra are too cumbersome to perform for routine
diagnosis, hence a semi-quantitative culture is performed by calibrated loop method.
A loopful of well-mixed uncentrifuged urine is inoculated on to CLED
agar/MacConkey agar and Blood agar without sterilizing the loop in between.

What is significant pyuria?


Presence of at least 1000 pus cells per ml of uncentrifuged urine is significant pyuria.
Ordinarily, presence of ≥10 pus cells/HPF in centrifuged urine and ≥5 pus cells in
uncentrifuged urine is considered significant. Some authors consider counts as low as
2-5 WBCs /HPF important in a centrifuged specimen in the female with appropriate
symptoms. In women, contamination from vagina may introduce large numbers of pus
cells into a sample of voided urine. The presence of squamous epithelial cells along
with pus cells in the sample is evidence that contamination has occurred and the pus
cell count is not significant.

What is significant bacteriuria?


Since normal voided urine tends to get contaminated with normal flora of the distal
urethra, differentiation of contamination from urinary tract infection is made by
quantifying the bacterial growth. Significant bacteriuria is a concept put forth by Kass
EH, who stated that there should be at least 1,00,000 bacteria of single type per ml of
urine. This count may not be applicable in all situations. Recent studies suggest that a
count of 102 per ml in acutely symptomatic women and a count of 10 3 per ml in
symptomatic men may be significant. Any growth obtained from urine collected via
suprapubic aspiration is significant. Lower counts may be significant when S. aureus
is the pathogen.
How is semi-quantitative culture performed?
A loopful of well-mixed uncentrifuged urine is inoculated on the agar medium
without sterilizing the loop and incubated at 37 oC overnight. Following incubation,
the number of colonies of single type is counted. A bacteriological loop of 3 mm
diameter approximately carries 0.001 ml of urine. If this amount of urine gives rise to
at least 100 colonies then the numbers of bacteria present in 1 ml can be obtained by
multiplying by 1000, i.e 1,00,000 per ml.

What is your observation?


Wet mount of urine shows plenty of pus cells and RBCs but few squamous epithelial
cells. More than 100 colonies of pink coloured (lactose fermenting), smooth, low
convex, circular colonies of a single type is seen on MacConkey's agar. Gram
stained smear of the colony shows gram negative bacilli, hanging drop shows
motile bacilli, and catalase test is positive. Results of biochemical reactions
are positive indole test, negative urea hydrolysis, negative citrate utilization,
positive MR test and negative VP test. TSI agar shows acid slant/acid butt with little
gas but no H2S. The isolate identified as Escherichia coli was obtained in significant
count.

What factors must be borne in mind while interpreting urine culture reports?
Urine in the bladder is sterile, small numbers of bacteria get into the urine from the
distal part of urethra while voiding. In typical cystitis, most often urine culture are
unimicrobial, recovery of more than one type of bacteria in urine indicates
contamination. Bacterial counts in the range of 10 2-103 in the absence of pyuria and
other symptoms usually indicates contamination. Rarely, mixed infection by more
than one type can occur; in such situations a repeat culture with same results is
reliable. Significant bacteriuria may not be applicable for suprapubic aspirated urine.
Bacterial counts can be lower if the patient is on antibiotics or has consumed large
amount of water before voiding urine. Bacterial counts can be higher if there is a long
delay between urine collection and culture.

What is sterile pyuria?


Presence of plenty of pus cells in urine but lack of growth on culture is sterile pyuria.
The reasons for this condition include recent administration of antibiotics, UTI by a
fastidious/auxotropic/anaerobic bacteria, urethritis due to gonococci,  non-gonococcal
urethritis (due to Ureaoplasma, Chlamydia, Trichomonas, or viruses) or renal
tuberculosis.

What is baceriuria without pyuria?


It is the presence of large numbers of bacteria in the urine but lack of significant
numbers of pus cells. This condition is usually seen in pregnant women where there is
retention of urine or when voided urine is held for a long time before culture.
How is this condition treated?
Trimethoprim-sulfamethoxazole for 3 days is considered the current standard therapy
for bacterial cystitis. Fluoroquinolones such as norfloxacin also works well.
Antibiotics should be selected on the basis of susceptibility testing of the isolate.

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