STUDY OF ISOLATION AND ITS ANTIMICROBIAL SUSCEPTIBILITY

OF GRAM NEGATIVE BACILLI IN URINE SAMPLE

. E. Navya sree
121317057008
INTRODUCTION :
A urinary tract infection (UTI) is an inflammatory process occurring in the kidney, ureter, bladder, or
adjacent structures when microorganisms enter through the urethra. Urinary tract infection is the most
commonly observed among clinical cases. Urinary tract infections are 35% among all hospitalized
patients.

The most commonly observed pathogens that cause urinary tract infections are from enterobacteriaceae
family. Most frequent cause of urinary tract infection is the E. coli although some other gram negative
bacteria also present in these infections. Other isolated pathogens of gram negative bacilli from urinary
tract infections patients are Klebsiella, Proteus, Pseudomonas. There are many factors that help in the increase
of urinary tract infection. The major factors include are sex, age, kidney tumors, pregnancy, stone,
catheterization, neurological diseases, urethral structures suppressed immune system, congenital/acquired
anomalies of bladder, enlarge prostrate, diabetes mellitus etc . Resistance to most of the antibiotics is still a
major problem.
.
Upper tract infections involves the kidneys ( pyelonephritis) , and lower tract infections involves the bladder
(cystitis) , urethra (urethritis) , and prostate ( prostatitis) .
AIM’S & OBJECTIVES:
1. To study the Isolation of Klebsiella gram negative bacilli from
urine sample.
2. To study the Antibiotic Susceptibility pattern of these Isolates.
3. To perform the test using Vitek 2 compact machine.
REVIEW OF LITERATURE :

AUTHOR YEAR WORK DONE

Waqas ahmed, adeel

saltar et al.,
2012 Study on antimicrobial susceptibility of gram negative bacilli
 

S.A.Lakshminarayana

et al., 2014 Antimicrobial resistance of pathogens associated with Urinary tract infections

Alfredo Ponce-de-Leon et 2009 The study of monitoring Antimicrobial resistance of bacteria from urine sample.
al.,

Devananda Prakash 2012 To determine the distribution and Antimicrobial susceptibility of uropathogens

et al.,

Mansi Agarwal et al., 2018 Gram negative bacilli causing hospital acquired infections and antibiotic suspectibility

Gerardo E. Guillen nieto 2019 Bacterial infections and their antibiotic resistance pattern
et al.,

Abbas mihankhah et al., 2013 Prevalence and antibiotic resistance pattern of bacteria isolated from urinary tract infections
Symptoms of UTIs
• A burning feeling when you urinate
• A frequent or intense urge to urinate, even though little comes out when you do
• Pain or pressure in your back or lower abdomen
• Cloudy, dark, bloody, or strange-smelling urine
• Feeling tired or shaky
• Fever or chills (a sign the infection may have reached your kidneys)
These symptoms may vary from mild to severe and in healthy women last an average of six
days. Some pain above the pubic bone or in the lower back may be present.
TRANSMISSION

Urinary tract infection can be transmitted by sexual intercourse, however, it can also be caused by bacteria
from the bowels. UTI is common among women. This is because the opening of the urethra is very close
to the vaginal and anal openings. There is a great chance that bacteria from your bowels will be spread by
wiping from anus to vagina after using the bathroom. Also, women tend to have much shorter urethras,
allowing bacteria to reach the bladder much easier. This is where the infection process begins. it will
infect the bladder and may spread to the kidneys. About 8-10 million people become infected with this
disease every year. About 80-90% of UTI's are caused by the Escherichia coli (E. coli), bacteria that is
found in the rectum. Bacteria can spread through person-to-person contact (for example, contaminated
hands of healthcare personnel, or other people via patient to patient) or, less commonly, by contamination
of the environment; the role of transmission directly from the environment to patients is controversial and
requires further investigation. Patients in health care settings also maybe exposed to bacteria when they
are on ventilators, or have intravenous catheters or wounds.
ANTIMICROBIAL SUSCEPTIBILITY :

Resistance to antibiotics is increasing day by day due to wrong or overuse. Major cause of urinary
tract infections are bacteria while best treatment for urinary tract infection is use of antibiotics.
Cephalosporins, broad spectrum Floroquinolones and Aminoglycosides are antibiotics which are used
in routine treatment of UTI. The best antibiotic used to treat the infection against gram negative
organism are cephalosporin. Resistance against bacterial pathogen is a worldwide issue. It varies
according to geographical and regional location. Antibiotic susceptibility pattern to urinary tract
pathogens varies which result in increase in resistance to most of antibiotics. Distribution of
uropathogens and their susceptibility to commonly used antibiotics varies from country to country.
MATERIALS AND METHODS

Methods used to collect urine sample :

Clean catch
Midstream catch
Catheter catch

CLED Agar for processing urine sample in the lab :

Composition :

Lactose-10gm
Enzymatic Digest of Gelatin-4gm
Enzymatic Digest of Casein-4gm
Beef extract-3 gm
L -Cystine-0.128 gm
Bromothyml Blue -0.02 gm
Agar – 15 gm
 MATERIALS USED ARE :

A BIO SAFTEY CABINET VITEK2 COMPACT

.
GENDER WISE PERCENTAGE
       
ISOLATED SPECIES TOTAL MALE % FEMALE %
 

E.COLI 65 34 /65 ( 54%) 31/65 (47%)

KLEBSIELLA 26 15/26 (57%) 11 /26 (42%)

PSEUDOMONAS 8 7 /8 (87.5%) 1 /8 (12.5%)

PROTEUS 4 3 /4 (75%) 1/4 (25%)

AGE WISE PERCENTAGE
 
AGE (YEARS) NO.OF PATIENTS MALE FEMALE

20 - 30 11 7 4

31 - 40 7 2 5

41 - 50 12 5 7

51 - 60 19 6 13

61 - 70 26 17 9

71-80 22 18 4

81 - 90 6 4 2
CLEAN CATCH / CATHETER
CATCH
  CLEAN CATCH CATHETER CATCH
ORGANISM

MALE FEMALE MALE FEMALE

 

E.COLI 29 / 55 26 / 55 5/10 5/10

 

KLEBSIELLA 12 /20 8/20 3/6 3/6

 

PSEUDOMONAS 2 /3 1/3 5/5 0/5

PROTEUS 1 /2 1 /2 2/2 0/2

RESULT :
Escherichia coli and klebsiella species have been reported to be the most common organisms causing UTI.
The study on gram negative bacilli was carried on specimens of 900 patients suspected of Urinary tract
infection of which 103 (11.4%) are positive. There were 23 inpatients and 80 out patients. The samples
were collected using clean catch and catheter catch. Males with 57.2% were observed to have higher cases of
UTI than the females 42.7%. as the significant growth recorded.
The age group of 61-70 years with 26 had the highest infection rate of against the other groups used.
Escherichia coli was the predominant uropathogens with 65 (63.1%) of total other microorganisms, followed
by klebsiella pneumonia 26 (25.2%) and pseudomonas aeruginosa and proteus mirabilus with 8 (7.7%) and 4
(3.8%) respectively.
The antimicrobial susceptibility pattern of gram negative bacilli isolated in the study revealed colistin and
amikacin to be most promising drug of choice for the treatment of Urinary tract infections. Colistin,
amikacin, ertapenem, nitrofurantoin are more susceptible to E.coli, klebsiella and pseudomonas. Amikacin,
gentamycin and ceftriaxone are susceptible to proteus.
Conclusion :
In this study a total of 900 urine samples from different patients of different age groups were
studied, out of which 897 were sterile cases. In the remaining samples E.coli, klebsiella,
pseudomonas, and proteus were identified.

The antibiotic sensitivity profile was obtained for each organism from individual sample and
appropriate treatment was provided.

The occurrence and infecting pattern of the organisms was studied and was found to be more in the
patients of higher groups(above 51).

From the results it was found that E.coli was most commonly effecting organism among the
patients of UTI.
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