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Final Presentation
Final Presentation
Final Presentation
Trioxide Bioaccumulation on
Melanophore Index in Channa punctata
Background - Max Berman
Experimental Design - Gehad Abdelaal
Results - Kailyn Broughton
Discussion - Shrey Mittal
Limitations - Tayler Blair
Research Question
How does arsenic trioxide bioaccumulation in
Channa punctata affect dermal chromatophores,
specifically dermal melanophores, and how can
this information be used as a biomarker for
arsenic pollutants in aquatic environments.
Background
● The expansion of industry and the advancement of human civilization has brought with it
increases in pollution, namely wastewater runoff filled with pesticides, toxins, and heavy
metal pollutants.
● Heavy Metal pollutant arsenic- Arsenic trioxide is one of the most abundant forms of
arsenic in nature and it is heavily used in both the manufacturing and agriculture
industries (Lin et al., 2014).
○ Levels of soluble arsenic have beens shown to be above permissible levels of 0.010 mg/L,
and this is a global occurrence (Kumari et. al, 2016).
● Fish chromatophores, in particular, were shown to have been impacted by the pollutants
in the water, with changes in dispersal being detected (Kaur and Dua, 2011).
● Channa punctata is a common species that can be used as a model system in various
pollution studies because they are readily available and suitable to evaluate the effects of
the toxicity of pesticides.
○ Channa punctata does not have the ability for robust physiological color change, such
as in other teleosts like Fundulus heteroclitus, they undergo frequent morphological
color change as they develop.
Background
● Color change is mediated by dermal chromatophores, including dermal melanophores.
There are two main methods for color change in teleosts:
○ Morphological- long-term color change affected by hormones that change chromatophore
density and form (Sugimoto, 2002).
○ Physiological- rapid color change controlled by neurotransmitters utilizing motor proteins
and cytoskeleton intracellular tracks (Ligon and McCartney, 2016)
● Biomarkers refer to an organism or tool that can be used to show the relationship
between changes in cellular responses, and environmental pollutants or toxic effects.
○ Previous research used Betta splendens chromatophores as cytosensors for various
pollutants and toxins (Chaplen et al., 2002).
Research Objectives:
● Establish how Channa punctata dermal chromatophores bioaccumulate arsenic.
● Observe how melanophore index is affected by arsenic trioxide bioaccumulation in
dermal melanophores
● Extend this information to another model organism, Fundulus heteroclitus, in order to
generate another biomarker and examine the impacts of arsenic bioaccumulation in
other ecosystems
Experimental Design
Hypothesis: Arsenic trioxide bioaccumulation for prolonged exposure leads to structural changes
in the dermal melanophores in Channa punctata making them a model organism to use as a
biomarker for arsenic pollution.
Experimental Design
Independent Dependent
Variable Variable
Duration of Level of overall
exposure to 1 mg pigment dispersion
1-1 of arsenic according to
trioxide (7, 14, 30, Hogben MI
60, 90 days)
Experimental Controlled
Control Group Group Variables
20 fish not 4 fish per group - Weight of Channa punctata : 35
+/- 5 g
exposed to exposed to arsenic
- Temperature of water in tanks :
arsenic for any for 7, 14, 30, 60, 25 C
time split and 90 days - Number of feedings/day : 1
randomly into 5 Type of food : Kijaro Japan
groups to serve as - Number of fish used
- Fish kept in individual tanks
parallel controls
Methods
● Before any treatment was given, the fish were kept under laboratory conditions for two
weeks
● In order to estimate the arsenic concentrations in the fish scales, the removed scales
were digested in concentrated nitric acid that was diluted by double distilled water.
○ A 1.0g scale sample was digested in 10 mL of nitric acid at 80℃ for one hour.
After the hour, 2 mL aliquots of the digested scale resultant were tested for As3+.
This was done by using hydride generation at a pH of 6.0, with sodium
borohydride being used as the reducing agent. The resultant hydride was
collected on a column and tested through coupled plasma emission
spectroscopy. The samples were tested at each duration level this way and the
spectroscopy was performed at USIC Indian Institute of Technology.
● Scales from both experimental and control groups were isolated and stained
with borax carmine and examined under a microscope
● The melanophore index was then calculated using the Hogben Method (Hogben
1942).
Results