SYNOPSIS PRESENTATION NEW EDITED

1. Department OF Fisheries and
Aquaculture UVAS, Ravi Campus,
PATTOKI
 Presented By: Muhammad Boota
(2020-mphil-1627)
Genetic Identification and
Phylogenetic Association of
Pangasius and Clarias species
with other Fauna of Pakistan

2. CONTENT
Background study
Objectives
Data analysis
Introduction and
need of study
Material and
method
Possible
outcome

3. Supervisory Committee:
Member
Member
Supervisor Dr. Shahid Sherzada
Dr. Farzana Abbas
Dr. Ali Hussain

4. FAO Report on Fisheries and Aquaculture, 2020
Aquaculture production attained another all-time record high of
114.5 million tones in live weight in 2018 and the production of
farmed aquatic animals has grown at an annual average of 5.3%
between 2001 and 2018. This reflects how total fish consumption
has been increasing over the last half-century, rising at a faster rate
than annual population growth. It is predicted that, by 2030, 62% of
global aquaculture production will be composed of freshwater
species such as carp and catfish as well as higher-value species
such as salmon, trout, and shrimp (FAO, 2020)

5. WORLD AQUACULTURE AND FISHERIES SCENARIO

6. Contribution of Capture Fisheries vs
Aquaculture to human Consumption

7. • Protein
• Iodine
• VitaminA,D
and B12
• Iron
• Omega 3
fatty acida
• Brain
growth
• Heart
attack
Strokes
• Auto
immune
disease
• Asthma
• Minerals
such as
calcium, zinc •
selenium
IMPORTANCE OF FISH

8.  External
features are
focused,
 such as
morphometric
and
 meristic
characters.
 Cytology
 Genetic
 Physiology
 anatomy are focused.
Based on protein, DNA
and RNA.
Traditional
taxonomy methods Modern
taxonomy methods

9. Works
with all
stages
of life

10. Identification
and products
or fragments
of an
organism

11. Food
Safety
and
accuracy

12. DNA Barcode
The method of
species
identification on the
basis of:
 short section of
DNA from a gene
or genes.

13. Nuclear DNA
markers
SNP
RFLP
RAPD
Microsatellite
DNA
NUCLEAR AND MITOCHONDRIAL DNA
MARKERS
Mitochondrial DNA
markers
Cytochrome D
loop
Cytochrome b
gene
Cytochrome c
gene

14. Offspring
cell
 It is derived from
 both parents. That is
why
 it is diploid.
 It has both introns
and exons.
 It has recombination
due to crossing over
Mitochondrial DNA
has genome from
 Only mother.
That is why
 it is haploid
 conserved.
 It has multiple
copies
 lacks introns and
recombination.
WHY MITOCHONDRIAL DNA IS PAREFERRED?
Nuclear DNA

15.  Mitochondrial DNA is circular,
double helical,
 without introns
 16500 nucleotides
 37 genes. Out of which
 13 genes encodes for
respiratory proteins
 and remaining 24 specify
 non coding RNA(22 tRNA, 2
rRNA )
MITOCHONDRIAL DNA

17. These are present on
the inner membrane
of mitochondria.
Cytochrome c has
life- supporting
function and are
important
component of
respiratory chain.
CYTOCHROME C GENES

18. Catfishes belongs to:
 order siluriformes
 having 39 families
 477 genera and
 3920 species.
• Theyarecalled
catfishes due to
presence of barbells.
 Cultivated in temperate
environment .
 About 107 species of
catfishes are present in
Asia and Europe.
 They feed on
phytoplankton,
zooplankton or nektons.

19. Clarias has been found to be paraphyletic.
(Mwita and Nkwengulila 2008).
They are found in inland waters
throughout much of the Old World, and
are one of the most widespread catfish
genera in the world (Ng and Heok 2003).
The genus is found in Southeast
Asia and East Asia westwards through
India and the Asia Minor to Africa (Lim et
al. 1999).

20. Clarias species are recognized by their
long-based dorsal and anal fins, which give
them a rather eel-like appearance. These
fish have slender bodies, a flat, bony head,
and a broad, terminal mouth with four pairs
of barbels. They also have a large,
accessory breathing organ composed of
modified gill arches (Ng and Heok 2003).

21. Also, only the pectoral fins have
spines (Sudarto et al. 2004).
Many of the species are of great
economic importance in
both fisheries and fish culture
(Teugels et al. 2001).

22. Among other cultured species
pangasius is the 4th most commonly
cultured species in the world after
salmon, shrimps and tilapia. The
tremendous potential of the
pangasius sector directs the
attention of world fisheries market
(Hekimoğlu et al. 2014).

24. Statement of Problem:
Proper identification of this fish fauna
of Pakistan is vital for its management
and conservation.

25. HYPOTHESIS :
cytochrome c gene might be a crucial
marker to determine phylogenetic
position of Clarius and Pangasius
species among other freshwater fish
fauna of Pakistan.

26. Research objectives :
 To identify and discriminate Clarias and
Pangasius species by using mitochondrial
DNA markers
 To evaluate genetic differentiation
between Clarias and Pangasius species.
 To determine the phylogenetic position of
Clarias and Pangasius species among
other freshwater fish fauna of pakistan.

27. Materials and Methods

28. SAMPLE SITE AND SAMPLE SIZE
• Sample size will be 75 and 25
sample will be taken from each site.
Kasur
SHEIKHUPURA
Muzaffargarh

29. METHODOLOY
DNA extraction
SequenceAnalysis
PCR/DNAamplification Sequencing
Gel electrophoresis
• Work will be performed in Aquaculture and
Fisheries lab. UVAS, Ravi Campus pattoki

30. a- Sample
preparation
b- Lysis
c- Protein Removal
d- DNA Binding
e- Wash
f- DNA Elution

31. Gel electrophoresis

32. Cyt c
Fish F1 TCAACCAAACCACAAAGACATTCCCAC
Fish RI TAGACTTCTGGGTGGCCAAAGAATCA
Primer selection :
Reported primer have been taken
from Ward et al, 2005.

33. PCR
Components
and Process

34. Sequencing and its analysis
Sequence
Align
UPLOAD IN
BOLD AND
OTHER
DATABASE
CONVERT
TO BARCOD

35. Data analysis
 Software used for data
analysis are:
 BIO EDIT
 BLAST
 MEGA 6
 DNASP and DNA STAR

36.  Morphological documentation
 Phylogenetic relationship
of Clarias and Pangasius species.
Possible Outcomes

37. 2021
2020
2020
2018
2018
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39. ?