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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Bayou Lafourche has 106 CFU/ml Aerobic bacterial load (Nathaniel, R. unpublished)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.
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
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.