Stuti K.

Desaia, Anup Padmanabhana, Sharvari Harshea, Ronen Zaidel-

Barb, and Linda J. Kenney
SALMONELLOSIS Symptoms
Salmonella typhimurium
Blood in the stool
meat
Diarrhea
egg
Abdominal cramps
milk
Nausea

water Fever
 small intestine
Innate Immune System
Methods of Infection

body’s first line of Mast Dendritic

defense cell cell

stomach
 Salmonella Pathogenicity Islands (SPI)

SPI 1 gene SPI 2 gene

invasion survival
 Planktonic Biofilm

Unphosphorylated SsrB, a transcription factor in

Salmonella Typhimurium is responsible for biofilm
formation.
Caenorhabditis elegans: Heterologous host
To determine if Salmonella exists as sessile aggregates in the persistently
infected C. elegans gut.

To understand the nature of Salmonella aggregates in the intestinal lumen

To investigate how the ability of Salmonella to form biofilms affect host survival.
 To determine the presence of Salmonella
aggregates in the intestinal lumen of C. elegans

Infection of C. elegans larvae

STm SsrB null E. coli

Observe

Samples were excited

Figure 1. SsrB-dependent formation of aggregates.
Image aquisition 2 4 6
E. coli

Spatial resolution STm

SsrB null
 To understand the nature of Salmonella aggregates in the intestinal lumen

Time-lapse projection of worm’s intestine

Differences in the Measurement of

intestinal luminal width aggregates in worms
 To visualize the ultrastructure of Salmonella
aggregates in C. elegans intestine

Fixed Stained

Embedded Washed

Sectioned Suspended

Incubated Transferred

Washed Embedded

Analyzed Intestine of STm infected C. elegans

 To investigate how the ability of Salmonella
to form biofilms affect host survival.

Percentage Survival of infected C. elegans

Exposure of C. elegans larvae

STm SsrB null E. coli

Plated

Counted Increased
Dormant Host Survival
Biofilm
Replicated formation Reduced
Infectious Host Survival
 To investigate the molecular basis for rapid
death in worms infected with the SsrB- null
mutant

Isolated SPI 1 Gene Log FC (ssrB null vs. STm)

Fliz 3.09

Eluted hilD 1.90

hilA 2.70
invF 2.13
Quantified
sptP 2.28

Fold-change in expression of SPI-1 genes in two-day

Treated old macrocolonies of STm and ssrB null

PCR analysis Shorter lifespan

SPI 1 gene
RNA sequence analysis Longer lifespan
 To measure the transcript levels of the
SPI 1 virulent genes

Day One Day Six

Reverse transcription

cDNA amplification

Plotted
RT-PCR analysis on SPI 1 virulent genes in SsrB-null
during in vitro biofilm condition

=
Replicated Faster
SPI 1 gene death
 To determine how ERK affects host innate
immunity

Protein separation
a-phospho-SEK-1
(ERK homolog)
Electro-transferred
a-tubulin

Blocking Levels of a-phospho-SEK-1 (ERK Homolog) in C. elegans

Primary Antibody Incubation

Low ERK ON Host Innate
level Immunity
Secondary Antibody Incubation SptP
High ERK Host Innate
level Immunity
Signal detection OFF
 Wild type-STM SsrB null strain
Biofilm Present
Absent
Virulent
down regulated
genes Up regulated
Host Innate
Activated Inctivated
Immunity
Host
Prolong Reduced
Survival

Activation of innate immunity in the presence of STm biofilms

Salmonella aggregates in the C. elegans intestine are static and vary
in size
Formation of biofilms down-regulates virulence.

Planktonic lifestyle is more detrimental to the long-term survival of

Salmonella.
Biofilms Promote Innate Immunity.

Salmonella biofilms enable dormancy to prolong in vivo existence.

Thus, biofilm formation is a survival strategy in long-term infection.
THANK YOU