In Klebsiella pneumoniae is a Gram-negative, nonmotile, rod-shaped

                 In the last decade, along with the problem of nosocomial infections,
multidrug-resistant bacteria in community and hospitals have soared. High
frequencies of multidrug-resistant bacteria have been grouped under the acronym
ESKAPE: Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobacter spp. The ESKAPE pathogens
are responsible for the majority of nosocomial infections and capable of ‘escaping’
the biocidal action of antimicrobial agents.

                Enterococci
are opportunistic bacteria which cause severe infectious diseases. Enterococcus faecium can cause several
infections including septicaemia, urinary tract infections, endocarditis, bacteraemia,
wound infections, meningitis, and neonatal sepsis. Acinetobacter are gram-negative coccobacilli that may cause
nosocomial infections in critically ill or debilitated patients’ particularly
ventilator-associated pneumonia and infections of the bloodstream, urinary
tract, and wounds. Klebsiella
pneumoniae is a Gram-negative, nonmotile,  rod-shaped bacterium .Klebsiella infections are seen mostly in people with
a weakened immune
system. The most common condition caused by Klebsiella bacteria
is pneumonia. In addition
to pneumonia, Klebsiella can also cause infections in
the urinary tract, lower biliary tract, and surgical wound sites. Enterobacter is
a genus of common Gram-negative, rod-shaped bacteria. These bacteria are pathogenic and cause opportunistic
infections in immunocompromised patients. The urinary and respiratory tracts are the most common sites of infection .Pseudomonas
is an opportunistic pathogen for humans lead to a broad spectrum of disease
such as urinary, burn, respiratory infections, and septicaemia. It is the
primary cause of ventilated, associated pneumonia in the intensive care unit

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This study is to consider the clinical importance of emerging of ESKAPE
pathogens in nosocomial infections to prepare feasible data about tracing and
treatment of infection related to ESKAPE pathogens that may be beneficial to
clinicians at the bed head. The awareness of residential antimicrobial
resistance can support selecting a convenient empirical therapeutic diet in
diseases due to ESKAPE pathogens.

Objectives

·        
To observe and analyze
the carriers of the ESKAPE pathogens in the ICU workers

·        
To develop innovative
therapeutic strategies.

·        
Study the environmental
factors leading to nosocomial infections in humans.

 

 

Methodology

               
Various organisms isolated from the nasal swabs will be tested for
ESKAPE pathogens along with their sensitivity pattern

Study
design and sample size

              
This will be a cross sectional study ,which will be carried out in two
months in the Department of Microbiology, in Sri Lakshmi Narayana Institute of
Medical Science ,Pondicherry .Clearance from the Institutional ethical
committee will be obtained . Nasal swabs will be collected from 80 health
workers from ICU after obtained informed consent. The health workers were not
taking any antibodies for last 2 weeks and were not admitted in hospital for
the last one year. Demographic details like age, occupation, sex, history of
underlying disease will be obtained

Collection
and transport of sample

              
Nasal swabs will be collected from both the anterior nose by swabbing
with a sterile swab, pre-moistened with sterile normal saline solution. Then these
swabs will be sending to the laboratory for culturing and testing the
antibiotic susceptibility. Antimicrobial susceptibility testing will be done on
Muller Hinton agar.

Processing
of sample

              After
receiving the sample in the laboratory, the swabs immediately inoculated on
Blood agar, chocolate agar, and macconkey agar. Then swabs are incubated at 37°C
for 24 hrs.

              Next
day, after overnight incubation, growths of bacterial colonies in all the
culture plates were noted. Identification of organisms was carried out using
standard microbiological methods, including Gram stain and other biochemical
reactions. In Blood agar, the type of haemolysis was noted. In MacConkey Agar,
Lactose fermentation was noted. By utilizing the lactose available in the
medium, Lac positive bacteria such as, Enterobacter
and Klebsiella will produce acid,
which lowers the pH of the agar below 6.8 and results in the appearance of pink
colonies .Non lactose fermenting organisms such as,
Pseudomonas aeruginosa produce mucoid
colonies which appear very moist and sticky. This phenomenon happens because
the organism is producing a capsule, which is predominantly made from the
lactose sugar in the agar.

            The
isolated colonies were Gram stained. After making out the gram reaction of the
organisms, appropriate biochemical tests were carried out to identify the
isolated organisms.

            Antimicrobial
susceptibility testing were done on Muller Hinton agar by  Kirby Bauer disc diffusion method a
semi-quantitative way based on diffusion ; small discs containing different
antibiotics, or impregnated paper discs, are dropped in different zones of the
culture on an agar plate, which is a nutrient-rich environment in which
bacteria can grow. The antibiotic will diffuse in the area surrounding each
tablet, and a disc of bacterial lyses will become visible. .Their susceptibilities
to antimicrobials will be recorded.

Implication

            Treatment of nosocomial infections
can be challenging due to the high level of antibiotic resistance of nosocomial
pathogens and the frequent immune depression of the host. We will use these
knowledge gathered on both microbial factors and host responses to develop
novel therapeutic strategies to prevent or fight nosocomial infections

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