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1
Bacteriophage
Maggie22
Nick
Hargett
2
Discovery
of
Maggie22
I
discovered
this
phage
at
the
Helen
S.
Ruder
Preserve
near
Fort
Mile
Creek.
The
area
was
grassy
and
had
plenty
of
life
around
it,
including
insects
and
other
plants.
The
sample
was
collected
near
the
base
of
a
plant
and
was
mainly
dry
soil
collected
1-1.5
inches
below
the
surface.
Temperature:
90℉
Humidity:
55%
3
4
Naming
of
Maggie22
I
named
the
phage
Maggie22
after
my
grandparent’s
new
puppy,
Maggie,
who
was
born
this
year.
Just
like
maggie,
I
figured
that
phages
are
also
small
and
cute.
5
Plaque
Morphology
Close-up
Plaque
morphology
is
clear
(lytic)
with
round
margins.
Entire
Plate
Plaque
are
mainly
medium
sized,
others
might
be
a
little
larger/smaller,
and
clear.
Calculated
titer:
12.6*10^10
pfu/mL
6
Restriction
Enzyme
Analysis
HaeIII:
1.)
395.20
bp
2.)
221.62
bp
3.)
112.85
bp
MseI:
1.)
11541.71
bp
2.)
7848.56
bp
3
bands
of
DNA
present
with
the
HaeIII
enzyme
2
bands
of
DNA
present
with
the
MseI
enzyme
1
2
3
1
2
7
Maggie22
vs.
Everyone
else
Resilience
Phage
lost
its
virulence
during
purification,
but
then
came
back
in
a
later
plaque
assay
EA
Cluster
The
restriction
enzyme
analysis
suggests
that
the
phage
belongs
to
the
EA
cluster
Sample
The
sample
was
mostly
dry
dirt
like
most
other
EA
cluster
m.
foliorum
phages.
Formed
2
morphologies
1st
uncontaminated
plaque
assay
included
both
clear
and
cloudy
plaque
8
Complications
Phage
lost
virulence
On
September
26th,
no
plaque
were
present
during
the
2nd
round
of
purification.
Contamination
Contamination
was
present
for
the
1st
round
of
purification,
however,
phage
was
still
able
to
infect
m.
foliorum.
Additionally,
although
not
immediately
present,
contamination
was
observed
in
the
plaque
for
the
high
titer
lysate.
(but
she
came
back)
9
Discussion
Results
that
echo
findings
in
publications
●
Phages
that
infect
the
genus
microbacterium
are
typically
members
of
the
EA
cluster.
●
Phages
with
greater
fitness,
like
reproductive
capabilities
and
high
titer,
are
more
likely
to
be
successful
during
purification
10
Future
Directions
What
other
roles
do
phages
have?
We
know
that
phage
are
viruses
and
kill
bacterial
cells,
but
do
they
have
roles
outside
of
killing
bacteria?
●
They
are
drivers
of
natural
selection
●
Regulate
bacterial
levels
11
Future
Directions
Phage
are
a
necessity
to
many
environments,
like
oceanic
environments
where
they:
●
Contribute
to
nutrient
availability
by
what
is
known
as
the
“viral
shunt”
In
the
human
gut
microbiome:
●
They
shape
the
bacterial
composition
of
the
intestines
by
infecting
and
lysing
certain
bacteria.
12
Resources
Jacobs-Sera,
Deborah,
et
al.
"Genomic
diversity
of
bacteriophages
infecting
Microbacterium
spp."
PLoS
One
15.6
(2020):
e0234636.
Nair,
A.,
Ghugare,
G.S.
&
Khairnar,
K.
An
Appraisal
of
Bacteriophage
Isolation
Techniques
from
Environment.
Microb
Ecol
83,
519–535
(2022).
https://doi.org/10.1007/s00248-021-01782-z
Brown,
Teagan
L.,
Oliver
J.
Charity,
and
Evelien
M.
Adriaenssens.
"Ecological
and
functional
roles
of
bacteriophages
in
contrasting
environments:
marine,
terrestrial
and
human
gut."
Current
Opinion
in
Microbiology
70
(2022):
102229.
10
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4
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Click to add new slide
Bacteriophage
Maggie22
Nick
Hargett
Discovery
of
Maggie22
I
discovered
this
phage
at
the
Helen
S.
Ruder
Preserve
near
Fort
Mile
Creek.
The
area
was
grassy
and
had
plenty
of
life
around
it,
including
insects
and
other
plants.
The
sample
was
collected
near
the
base
of
a
plant
and
was
mainly
dry
soil
collected
1-1.5
inches
below
the
surface.
Temperature:
90℉
Humidity:
55%
Naming
of
Maggie22
I
named
the
phage
Maggie22
after
my
grandparent’s
new
puppy,
Maggie,
who
was
born
this
year.
Just
like
maggie,
I
figured
that
phages
are
also
small
and
cute.
Plaque
Morphology
Close-up
Plaque
morphology
is
clear
(lytic)
with
round
margins.
Entire
Plate
Plaque
are
mainly
medium
sized,
others
might
be
a
little
larger/smaller,
and
clear.
Calculated
titer:
12.6*10^10
pfu/mL
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hargetne@miamioh.edu
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