Genetic identification of purebred and crossbred water buffalo [Bubalus bubalis (linnaeus, 1758)] in selected farms in Malaysia
Water buffalo or Bubalus bubalis is a domesticated animal that plays essential roles in agriculture, economy and food production. Water buffalo is classified into two sub- species based on the phenotypic, genotypic and cytogenetic. The two subspecies of water buffalo is murrah buffalo (Bub...
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| Main Author: | |
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| Format: | Thesis |
| Language: | English |
| Published: |
2019
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| Subjects: | |
| Online Access: | http://psasir.upm.edu.my/id/eprint/84070/1/FPV%202019%2015%20-%20ir.pdf |
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| Summary: | Water buffalo or Bubalus bubalis is a domesticated animal that plays essential roles
in agriculture, economy and food production. Water buffalo is classified into two sub- species
based on the phenotypic, genotypic and cytogenetic. The two subspecies of water buffalo is
murrah buffalo (Bubalus bubalis bubalis) and swamp buffalo (Bubalus bubalis carabensis).
Identification based on morphology is not enough to confirm the species or to make the comparison
between the two sub-species. The problem arises when these two sub-species mates and
produce their crossbreed offspring, which is indistinguishable from their parents.
Therefore, the aim of this study is to determine three sub-species of water buffalo available
in Pusat Ternakan dan Pembiakan Kerbau, Telupid, Sabah and Ladang Ternakan Kerbau Semenyih
(Semenyih Farm) using karyotyping and molecular methods. In addition, phylogenetic analysis is to
determine the differences in the nucleotide sequences, associated with the relationship
evolutionary events between species. The blood of 112 animals were taken, cultured, terminated and
harvested using conventional karyotyping protocol to obtain the chromosomes. Results showed
that from 112 buffaloes in PLadang Ternakan dan PembiakanHaiwan, Telupid, Sabah, 97 animals
successfully identified were confirmed as swamp buffalo presented with 48 chromosomes and the
remaining 7 were identified as crossbred buffalo presented with 49 chromosomes. However, in
Semenyih Farm, 8 animals were successfully identified as murrah buffalo presented with 50
chromosomes. In molecular work, the bloods were collected and the PCR were conducted to quantify
the d-loop of mitochondrial DNA in three sub-species. Two pairs of primers known as NP1 and NP2
were designed to amplify the full d-loop region by PCR. The purified PCR products from three
sub-species were then sent for sequencing. Results from sequencing showed that the full
d-loop of mitochondrial DNA is successfully amplified, consist of 1000 base pairs. Based
on the multiple sequence analysis, the three subspecies of water buffalo shared many conserve
region and they also showed some differences in their nucleotide sequences. The mean molecular weight (Da) is also differ each sub-species, which swamp with 343,747.70
Da, crossbreed with 345,331.30 Da and murrah with 335,374.22 Da. All d-loop
mitochondrial DNA sequences from this study hit the NCBI blast for the most similar
sequences with 99% identical to the sequences available from the database.
Phylogenetic tree was constructed using maximum likelihood analysis using software
MEGA 7.0. From the phylogenetic results, the swamp and murrah clearly formed two
different clades, showing that they are diverged from each other. On the other hand,
crossbreed buffalo is claded into the swamp buffalo, showing that swamp buffalo is
more dominant in the crossbreed compared to murrah buffalo. By combining
karyotyping method, molecular study and also phylogenetic tree, the result obtained
from this study is more convincing and complete compared to analysis done solely on
one aspect. In conclusion, the genetic identification is crucial in identifying the three
sub-species of water buffalo, as it can provide more concrete and clear result compared
to conventional method such as morphological analysis. |
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