Int.J.Curr.Microbiol.App.

Sci (2014) 3(3): 115-119

ISSN: 2319-7706 Volume 3 Number 3 (2014) pp. 115-119

http://www.ijcmas.com

Original Research Article

Isolation and antibiotic sensitivity of Klebsiella pneumoniae
from pus, sputum and urine samples

K.N.Ravichitra1*, P.Hema Prakash2, S. Subbarayudu3 and U. Sreenivasa Rao4

Department of Microbiology, Alluri Sitarama Raju Academy of Medical Sciences (ASRAMS),

Eluru - 534 005 West Godavari District, Andhra Pradesh, India
*Corresponding author

ABSTRACT

Gram negative pathogens are an important cause of community and hospital

acquired infections throughout the world. Klebsiella pneumoniae has become one
of the more common cause of these infections and one of the important aspects of
Klebsiella associated infections is the emergence of multi-drug resistant strains
Keywords
particularly those involved in nosocomial diseases. So the knowledge of the
resistivity pattern of Klebsiella isolates has been the global necessity. This study
Klebsiella was done to determine the isolation rate of Klebsiella, their antibiogram and for the
pneumoniae; presence of resistant strains from various clinical samples. A total number of 698
nosocomial pus, 312 sputum and 2176 urine samples were included in the present study.
infections; Isolates of Klebsiella were identified by standard microbiological techniques and
antimicrobial their antibiogram determined by Kirby-Bauer disc diffusion method. Of the 3186
drug resistance; samples processed 1798 (56.4%) samples were culture positive for various
ESBLs. organisms. A total of 1871 organisms were isolated, of them 480 (25.6%) were
Klebsiella pneumoniae. The frequency of ESBL producers in our study was 76
(15.8%) of all Klebsiella isolates. The present study reveals the frequency of
isolation of Klebsiella from various clinical samples and their tendency towards
antibiotic resistance.

Introduction

In 1883 Friedlander isolated a capsulated Klebsiella is a gram negative, non-motile,

bacillus from the lungs of patients who encapsulated, lactose fermenting,
had died of pneumonia (Patrick R Murray, facultative anaerobe belonging to the
2005). This was named after him as Enterobacteriaceae family (Elmer W.
Friedlander s bacillus. Later on this Koneman, 2006). It is the second most
organism was given the generic name of popular member of the aerobic bacterial
Klebsiella, which is ubiquitously present flora of the human intestine. It is the most
and reported worldwide. common causative agent of nosocomial
and community acquired infections.

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It has even replaced Escherichia coli in Microbiology. A total of 698 pus, 312
some centers as a nosocomial pathogen. It sputum and 2176 urine samples were
causes pneumonia, urinary tract infection, collected during this period with universal
other pyogenic infections, septicemia and safety precautions and were transported to
rarely diarrhea (Arti Kapil, 2013). the laboratory without delay. The pus
samples were either aspirated by
Biochemically typical strains of Klebsiella disposable syringes or collected onto
pneumoniae are resistant to a wider range sterile cotton tipped swabs. Sputum and
of antibiotics than are most Escherichia clean voided midstream urine was
coli strains. They are nearly always collected into screw top containers (Patrica
naturally resistant to ampicillin (Patrick R M. Tille, 2014). Samples were obtained
Murray, 2005). Resistance of Klebsiella to from both inpatients and outpatients, of all
previously sensitive antibiotics is also age groups and of both sexes.
increasing in the recent years due to
overuse and misuse of antimicrobial Characterization of bacterial isolates
agents and or natural causes.
Pus, sputum and urine samples were
Of particular concern is the Extended aseptically inoculated on to Blood and
Spectrum Beta Lactamase (ESBL) MacConkey agar plates and incubated
producing Klebsiella pneumoniae that overnight at 370C. Klebsiella isolates were
have been steadily increasing over the past identified by their morphology and
years and rapidly spreading worldwide biochemical characteristics. Morphology
that pose a serious threat for healthcare of Klebsiella identified were large dome
associated infections. Increasingly the shaped colonies on blood agar and lactose
ESBL Klebsiella pneumoniae are also fermenting mucoid colonies on
showing co-resistance to other MacConkey agar. In gram staining, gram
antimicrobial agents like quinolones and negative, short, plump, straight rods were
aminoglycoside antibiotics. Both seen .The biochemical characters
morbidity and mortality is increased when identified were positive Voges-Proskauer
infection is caused by these drug resistant test, positive citrate utilization test,
organisms. Antibiotic sensitivity pattern positive urease test, acid and abundant gas
may change from time to time and place to production from glucose, lactose, sucrose,
place. maltose and mannitol sugar fermentation
tests (Elmer W. Koneman, 2006; J.
Therefore updated knowledge of the drug Gerald collee, 2012).
resistance pattern in a particular region is
useful in clinical practice. This work gives Antimicrobial susceptibility testing
an account of isolation of Klebsiella from
clinical pus, sputum and urine samples, Was done for all the isolates on Mueller-
their antibiogram and presence of resistant Hinton agar plates by Kirby-Bauer disc
strains in various samples. diffusion method according to the CLSI
guidelines 2012 (CLSI, 2012). Reduced
Materials and Methods susceptibility to cefotaxime (30 g) and
Collection of samples Ceftriaxone (30 g) with zone sizes
27mm and 25mm respectively were
This study was carried from November used as screening method for ESBL
2012 to October 2013 at the Department of production.
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Results and Discussion in good agreement with Manikandan et al

(2013) followed by 29.2% in pus samples,
A total of 3186 pus, sputum and urine which is similar to Valarmathi et al (2013)
samples were processed during our study and 22.9% in urine samples which is in
period. 1871 bacterial isolates were agreement with R.Sarath babu et al (2012).
obtained from culture positive 1798 Most of the isolates in our study were not
samples. Of them 480 were Klebsiella multidrug resistant. The frequency of
pneumoniae (Table -1 & chart-1). The ESBL producers in our study was 76
sensitivity and resistance pattern of the (15.8%) of all Klebsiella isolates. Of them
Klebsiella isolates to various antibiotics in isolates from pus constituted the highest
our study is shown in the table -2. Of the 36 (23.1%) of all pus isolates, which is in
480 Klebsiella isolated, 76 (15.8%) were good agreement with reports from
ESBL producers, 36 isolates were from B.L.Chaudhary et al (2013), followed by
pus samples, constituting 23.1% of all pus 10 (14.7%) of sputum isolates which is
isolates, 15 were from sputum samples little higher than that reported by Ugur
constituting 22.1% of all sputum isolates Gonlugur et al (2004) and 30 (11.7%) of
and 25 were from urine samples all urine isolates which is similar to
constituting 9.8% of all urine samples. Shamweel Ahmed et al (2009). Our study
showed good sensitivity to imipenem,
Klebsiella species have been associated accounting for 94.9% of pus, 80.4% of
with different types of infections. sputum and 89.9% of urine samples.
However the main importance of Sensitivity to cefaperazone and sulbactam
Klebsiella as a pathogen is in causing was also good accounting to 81.1% of pus,
infections in hospitalized patients, the 88.2% of sputum and 89.1% of urine
strains responsible are nearly always samples. Sensitivity to quinolones and
biochemically typical members of aminoglycosides antibiotics was moderate
Klebsiella pneumoniae (Patrick R Murray, in our study.
2005).
The present study reveals the incidence of
Moreover, extensive use of broad infections due to Klebsiella and their
spectrum antibiotics in hospitalized tendency towards antibiotic resistance.
patients has led to both increased carriage Multidrug resistant bacteria are emerging
of Klebsiella and the development of worldwide which causes major public
multidrug resistant strains like those of health problems and challenges to health
Extended Spectrum Beta Lactamases care.
(ESBLs) (Archana Singh Sikarwar et al.,
2011). These multidrug resistant strains Knowledge about the common organisms
cause serious nosocomial and community associated with infections, the resistance
acquired infections that are hard to patterns of these bacterial strains in a
eradicate by using available antibiotics. geographical area will help to guide
Hence the need to determine the appropriate and judicious antibiotic use,
antibiogram of these pathogens in order to formulate antibiotic policies and for
evaluate the efficiency of empirical drug infection control intervention programmes.
treatments formulated in our hospitals. However only screening tests were
performed for detection of ESBLs in our
In the present study culture positivity for study, confirmatory studies are required
Klebsiella in sputum was 30.9% which is for further evaluation.
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Table.1 Total number of organisms isolated

Number of Culture % of culture

Sample Total no of samples
positives positives
Pus 698 482 69.1
Sputum 312 212 67.9
Urine 2176 1104 50.7
Total 3186 1798 56.4

Table.2 Antibiotic sensitivity pattern

Pus Sputum
Antibiotics Urine
R% S% R% S% R% S%
Amoxyclav 58.2 41.8 78.4 21.6 65.9 34.1
Ofloxacin 35.4 64.6 33.3 66.7 40.3 59.7
Cefotaxime 23.1 76.9 19.8 80.2 8.7 91.3
Ceftriaxone 20.6 79.4 22.1 77.9 9.8 90.2
Gentamicin 12.7 87.3 37.6 62.4 40.3 59.7
Cefoperazone&
18.9 81.1 11.8 88.2 10.9 89.1
sulbactam
Imipenem
5.1 94.9 19.6 80.4 10.1 89.9

Chart.1 Culture positivity of Klebsiella pneumoniae

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Acknowledgement Patricia M. Tille. 2014. Bailey & Scott s

Diagnostic Microbiology. 13th edition.
I thank the management of ASRAM St. Louis, Missouri : Mosby Elsevier
Medical College and HOD of publications.
Microbiology Department, Patrick R Murray, Barry Holmes, Hazel
Dr. S. Subbarayudu for allowing me to M. Aucken. 2005. Topley & Wilson s
conduct this short study. Microbiology & Microbial Infections.
Volume 2. 10th edition. Salisbury, UK:
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