Assumption of Using Blood Serum of Komodo Dragon for Antibacterial anctivity

1. SZABIST, Karachi

3. Research Published on
29th August 2012

4. Different Biologists Contributions:
• Mark Merchant
• Danyell Henry
• Rodolfo Falconi
• Becky Musher
• Judith Bryja

9. Taxonomy
• Phylum: Chordata
• Class: Reptilia
• Order : Squamata
• Family: Varanidae
• Species: Varanus Komodoensis

10. Features
• Largest lizard on Earth
• May live up to 100 years
• Can run 20 kilometers per hour
• Dive upto 4.5 meters ( 15 feets) deep in water
• Kills its prey with a lethal bite
• Believed to inject toxic substance into the
blood stream of the animals
• Saliva contains potent and complex venom

13. Primary goal of the research was to
characterize the antibacterial
properties of Serum from the
Komodo Dragon

15. Nutrient Broth
Low EEO Aggrose
Bacterial Cultures

16. Gram Negative
Bacteria

17. • Esherichia Coliform
• Shigella Flexnerri
• Klebsiella Oxytoca
• Salmonella Tryphimurium
• Providenica Stuartii.

18. Gram Positive
Bacteria

19. • Staphylococcus Aureus
• Streptococcus Epidermitis
• Streptomyces Griseus

20. Collection of Blood Sample
of Komodo Dragon

22. • Collected from komodo Dragons at the Houstan
and San Antonio Zoo.
• Collected from 3 adults of weight 20 – 81.5 kg
• Collected from 5 juveniles of weight 1.5-6.2 kg
• Drawn from the tail vein
• Transferred to vaccutainer tube
• Allowed to clot for 5 hours before collecting the
serum.
• Serum was pooled so that the average
antibacterial values can be generated
• Stored at -20’C until used for bacterial assay.

24. Serum Volume Dependent
Antibacterial Assays

25. • Took 200 samples of serum containing
volumes of 20-200 ul in micro-titer plates.
• The plates were then inoculated with 5ul of
bacterial culture.
• Then the samples were incubated for 24 hours
at 37’C.
• Optical density of each sample was measured
at 0,3,6,12 and 24 hour using Benchmark Plus
micro-titer plate reader at 430 nm.

26. Kinetic Antibacterial Assay

27. • 500 ul of Komodo Dragon serum was inoculated
with 20 ul of an E.Coli culture in log phase
growth.
• 50 dilution of each sample were spread onto the
surface of Nutrient Broth agar plates to
determine the CFU’s for each sample.
• To determine the possible antibacterial
mechanism, pooled serum was incubated with
pronase ( isolated from Streptomyces Griseus), 5
mm EDTA or 5 mm phosphate for 30 minutes at
ambient temperature.

29. Results of Serum
Volume Dependent
Antibacterial Assays

30. Results for Gram Negative Bacteria
• 10% of Komodoensis serum produced 44% decrease in
E.Coli growth.
• 25% of serum produced 58% decrease in E.Coli growth.
• 50% of serum poduced 75% decrease in E.Coli growth.
• 75% of serum produced 93% decrease in E.Coli growth
• 100% of serum produced 98% decrease in E.Coli
growth
• 1 ul of serum has the capacity to kill approximately
2.24 million of E.Coli bacteria.

31. • 25% of serum produced 65% decrease in
Klebsiella Oxytoca.
• 58% of serum completely inhibited the
growth of Klebsiella Oxytoca in 24 hours.
• 10% of Serum inhibited Shigella Flexneri and
Salmonella Typhimurium by 95% at 24 hours
• Providencia Staurti’s growth is completely
inhibited by 10% Komodoensis serum.

32. Results for Gram Positive Bacteria
• 10% of komodoensis serum inhibited the growth
of Staphylococcus Aureus by 64 % at 24 hours.
• 25% of serum inhibited 68% growth of
Staphylococcus.
• 50% of serum inhibited 84% growth of
Staphylococcus.
• 75% of serum inhibited 94% growth of
Staphylococcus.
• 10% of serum inhibited 49% growth of
Streptococcus Epidermitis

33. Results for Kinetic
Antibacterial Assay

34. • Antibacterial activity occurred quickly as 18% of
the bacteria (2.95 million) were killed in 5 mins.
• Further incubation resulted in 43% (7.05 million)
of the bacteria killed in 10 mins after the
exposure to serum
• In 20 mins 68% (11.2 million) of E.coli bacteria
were killed.
• Treatment of serum with 5 Mm EDTA or 5 Mm of
Phosphate only resulted in 6.3% or 8.7% of the
antibacterial activity.

35. • This activity of Serum suggests that Komodo
Dragon are resistant to Microbial Infections.
• It would be reasonable to expect the Komodo
Dragons would have developed Innate
Immune Mechanisms to combat possible
infections by the bacteria in their own Saliva.

36. • Before this study ,crocodilians were
considered to have the most potent serum
complement defense system.
• The result in this study show that the potency
of Komodo Dragon serum is much higher than
the crocodilians