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Neonatal Sepsis: Antibiotic Resistance

This document summarizes a study on cephalosporin and carbapenem resistance in gram-negative bacteria causing neonatal sepsis. The study aimed to detect resistant microorganisms using biochemical assays and analyze antibiotic susceptibility patterns, identify ESBL and carbapenemase production, and detect resistance genes. Biochemical tests identified E. coli isolates. Disc diffusion tests showed resistance to several antibiotics. Combination disk tests identified ESBL and carbapenemase production. PCR detected the NDM gene in some isolates. Further tests are planned to fully characterize resistance determinants and their transmissibility.

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Tamoghna Naskar
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129 views17 pages

Neonatal Sepsis: Antibiotic Resistance

This document summarizes a study on cephalosporin and carbapenem resistance in gram-negative bacteria causing neonatal sepsis. The study aimed to detect resistant microorganisms using biochemical assays and analyze antibiotic susceptibility patterns, identify ESBL and carbapenemase production, and detect resistance genes. Biochemical tests identified E. coli isolates. Disc diffusion tests showed resistance to several antibiotics. Combination disk tests identified ESBL and carbapenemase production. PCR detected the NDM gene in some isolates. Further tests are planned to fully characterize resistance determinants and their transmissibility.

Uploaded by

Tamoghna Naskar
Copyright
© Attribution Non-Commercial (BY-NC)
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Cephalosporin and Carbapenem resistance in gram negative bacteria causing Neonatal Sepsis

Under the guidance of Dr.Sulagna Basu Scientist C Dept. of Bacteriology, National Institute of Cholera and Enteric Diseases,Kolkata

Presented by Poulami Dutta Regn no. 1641110013

Neonatal Sepsis
Blood infection by bacteria or any other virulent pathogen is referred to as septicemia. Sepsis that develops within 28 days of life is known as
neonatal sepsis or sepsis neonatorum.

Neonatal sepsis is of 3 types:

Congenital neonatal sepsis Early onset sepsis(EOS) Late onset sepsis(LOS)

Clinical features
Hypothermia or fever Lethergy,poor cry,refusal to suck

Absent neonatal reflexes


Bradycardia/Tachycardia Respiratory distress & gasping respiration

Each year, globally around 3.6 million babies die within the first 28 days of life (the neonatal period) among which 98% deaths occur from developing countries (Zupan, 2005).

In India there is an estimated 1.84 million under five children death and of these approximately 41% happen in the neonatal period (Lahariya et al., 2010).
In India the major causes of neonatal deaths are sepsis and pneumonia 30.4%, birth asphyxia 19.5% and pre-maturity 16.8

Zupan J. 2005. Perinatal mortality in developing countries. N Engl Med. 352:20472048 Baqui AH, Darmstadt GL, Williams EK, Kumar V, Kiran TU, Panwar D.et al. 2006. Rates, timing and causes of neonatal deaths in rural India: implications for neonatal health programs. Bull World Health Organ.84:706713.

Antibiotics administered in case of neonatal sepsis


Gentamicin

Ampicillin

Cephalosporin
Carbapenem

Aminoglycoside

OBJECTIVE

Work Plan & Methods used Detection of the gram negative micro-organism using a few
biochemical assays

Phenotypic analysis 1.Antibiotic susceptibility patterns Disc diffusion test MIC determination 2.Detection of ESBLs and MBLs Combination disk test 3. Detection of the Carbapenemase Modified Hodge test

Genotypic analysis
1.PCR followed by agarose gel electrophoresis 2 Conjugation experiments to detect the transmissibility of carbapenem and cephalosporin resistance determinants

Results of the biochemical tests


Indole test Positive TSI test A/A Gas +,no H2S Citrate test Negative (green) Mannitol motility test Positive,yellow hazy Urease test Negative

The identified micro-organism is E.coli

Results of the disc diffusion tests


CAZ SXT 1 2 3 4 5 6 7 8 9 1 0 R R R R R R R R R R S R R R R R R R R R CRO CL R R R R R R R R R R S S S S S R R S R S FEP PIP R R R R R R R R R R R R R R R R R R R R AK R R R R R R R R R R GM CIP S R R R R R R R R R S R R R R R R R R R MEM GAT MI S R R R S R R R R R S R R R S R R R R R S S S S R S R S R S TGC TZP S I I S S S R S S R S R R R R R R R R R

Zone of clearance

Fig: Kirby -Bauer disc diffusion method of determining antibiotic susceptibility pattern of some representative test isolates (E.coli)

Graphical representation of the antibiotic susceptibility patterns of 10 test isolates

Results of the combination disk tests

Zone of clearance
Fig:Combination disk method of determining antibiotic susceptibility pattern of some of the representative test isolates (E.coli)

ESBL+ve
(CTX+C/CTXCC ) - CTX 5 (CTX+C/CTXCC) CTXCX 5

AmpC +ve
(CTXCX/CTXCC)- CTX 5 (CTXCX/CTXCC)- CTX+C 5 (MRP+BO/MRP+CL)- MRP5 (MRP+DP)-MRP 3

KPC+ve
(MRP+DP/MRP+CL) MRP 3 (MRP+BO) MRP 5

MBL+ve
(MRP +DP)-MRP 5

PCR results

NDM gene (775bp)

Work in progress
Further tests will be carried out to confirm the presence of the resistant determinants. Phenotypic tests like MIC determination tests and Modified Hodge tests. Genotypic analysis-screening of other genes. Conjugation experiments.

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