Post on 03-Feb-2016
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Antimicrobial Properties of Garfish Blood
Rachel E. Laird
Nicholls State University
Antimicrobial Studies Involving Fish Blood
Rainbow trout (Oncorhynchus mykiss) model.
Hollebecq et al;1995; - complement blood plays a significant role in defending against infection, the bactericidal activity of the trout blood is more accurate in defending against furunculosis.
Bandin et al.; 1995; - opsonized bacteria increased their survival inside phagocytes
Nikoskelainen et al.;2002; - rainbow trout complement activity increased from 10oC to 35oC
Why Gar Blood?• There have been no studies to date on the
effects of gar blood on bacterial activity.
• Help gain a better understanding of garfish biology.
• Establish a foundation to further explore blood components of an ancient fish species.
• Isolating and characterizing the molecules would help aid in a better understanding of host-pathogen interactions and pathways.
BackgroundFish immune responses against bacteria involve both cellular and non-cellular components
Complement and small antimicrobial peptides are thought to be important innate immune responses
Activity of blood is dependent upon such factors as temperature.
Bayou Lafourche has 106 CFU/ml Aerobic bacterial load (Nathaniel, R. unpublished)
Complement – series of enzymatic events resulting in formation of a ‘membrane attack complex, ‘death by a hole puncher’
Antimicrobial Peptides – cationic peptides, 5-60 aa, disrupt membrane integrity.
Objective:
To determine the antimicrobial properties in Garfish Blood.
Hypotheses:
• Gar blood will have a bactericidal effect.
• Bactericidal effect will follow a dose dependent and time dependent mechanism.
Audubon Bridge Water Collection Point
New OrleansThibodauxHouma
Satellite Image of Mississippi Drainage Basin
Methods
Control =Medium + Bayou water
Test = Medium + Bayou water + Blood
Blood Sterility Control = Medium + Blood
All results incubated at 37°C for 18 hrs, OD600 readings taken
Serial dilutions
Bayou Lafourche Water
Effects of Gar Blood on Bacterial GrowthB
acte
rial
Nu
mb
er C
FU
/ml
Bac
teri
al N
um
ber
CF
U/m
l
Mean Effects of Gar Blood on Bacterial GrowthB
acte
rial
Nu
mb
er (
CF
U/m
l)
N=4; *P<0.001
*
*
Effects of Gar Blood on Bacterial Growth over 24 HoursB
ac
teri
al N
um
be
r C
FU
/ml
N=6
Effect of Heat Treated Gar Blood on Bacterial Growth
Bac
teri
al N
um
ber
CF
U/m
l
Mean Effects of Heat Treated Gar Serum on Bacterial GrowthB
acte
rial
Nu
mb
er C
FU
/ml
N=3
Summary
Gar blood (@10%) showed antimicrobial activity against up to ~1,000 bacteria/ml
We did not yet find antimicrobial activity to follow a time dependent mechanism.
We have not yet been able to show antimicrobial activity to be heat labile.
Acknowledgements
Dr. Quenton FontenotDr. Allyse FerraraMr. Rickey Verrett (Gar Fisherman)Ms. Olivia SmithDr. Rajkumar Nathaniel
Funded by:LA Board of Regents