1.Identification of Newcastle Disease Virus sub-genotype VII 1.1 isolated from chickens in Sabah, Malaysia
Syamsiah Aini, S. ; Leow, B.L. ; Faizul Fikri, M.Y. ; Muhammad Redzwan, S. ; Faizah Hanim, M.S.
Tropical Biomedicine 2022;39(No.4):579-586
Newcastle disease (ND) is an extremely contagious and fatal viral disease causing huge economic losses
to the poultry industry. Following recent ND outbreaks in Sabah in commercial poultry and backyard
farms, it was speculated that this could be due to a new introduction of Newcastle Disease Virus (NDV)
genotype/sub-genotype. Here we report the genetic characterization of NDVs isolated from Sabah during
early 2021. All isolates were amplified and sequenced with primers specific to the viral fusion (F) gene
using reverse transcription-polymerase chain reaction (RT-PCR). Nucleotide sequence analysis of the F
gene showed that all isolates shared similar homology of 99.4% with NDV strain from Iran isolated in
2018. Amino acid sequences of the F protein cleavage site revealed the motif of 112RRQKRF117 indicating
all isolates were of virulent strain. Phylogenetic analysis demonstrated that all isolates were clustered
under sub-genotype VII 1.1 and clustered together with isolates from Iran (previously known as subgenotype VIIl). The present findings suggested that there is an emerging of a new sub-genotype into
the poultry population in Sabah and this sub-genotype has never been reported before in Malaysia.
Therefore, transboundary monitoring and continuous surveillance should be implemented for proper
control and prevention of the disease. A further molecular epidemiological analysis of NDV is needed
to well understand the circulatory patterns of virulent strains of NDV in the country to prevent future
outbreaks.
2.Molecular characterisation of hemagglutinin and neuraminidase genes of the first highly pathogenic Avian Influenza H5N1 2.3.2.1c virus isolated from Sabah, Malaysia
Leow, B.L. ; Syamsiah Aini, S. ; Faizul Fikri, M.Y. ; Muhammad Redzwan, S. ; Ong, G.H. ; Faizah Hanim, M.S.
Tropical Biomedicine 2022;39(No.2):160-169
Highly Pathogenic Avian Influenza (HPAI) is a highly contagious disease in poultry. The outbreaks can
lead to flock mortality up to 100% in two to three days. In July 2018, high mortality in a commercial
layer farm in Kauluan village, Sabah was reported. Samples were sent to Veterinary Research Institute
Ipoh for diagnosis. Virus isolation and molecular detection is carried out simultaneously. The causative
agent was then identified as AI H5N1 virus by real time reverse transcription-polymerase chain reaction
(RT-PCR). The virus was then subjected for further nucleotide sequencing of full length hemagglutinin
(HA) and neuraminidase (NA) gene. The PQRERRRKR/GLF motif at the HA cleavage site indicated that the
isolate was of HPAI virus. Phylogenetic analysis of the HA gene showed that the isolate was belonged to
the clade 2.3.2.1c virus. In the HA gene, besides the S133A substitution, the virus possesses conserved
amino acid at most of the avian receptor binding sites including the glutamine (Q) and glycine (G) at
position 222 and 224 respectively, indicating that the virus retains the avian-type receptor binding
preference. As such, the zoonotic potential of the virus was relatively low. On the other hand, though
the N154D and T156A substitution were detected in the same gene, the pandemic potential of this
Sabah 2.3.2.1c virus is low in the absence of the Q222L, G224S, H103Y, N220K and T315I. A typical
20 amino acid deletion with loss of four corresponding glycosylation sites in the NA stalk region was
visible. Though three NA resistance markers were detected, the virus was predicted to be sensitive
to NA inhibitor. This is the first HPAI H5N1 outbreak in Sabah. The introduction of this virus into East
Malaysia for the first time raised an alert alarm of the future epidemic potential. Strict farm biosecurity,
continuous surveillance programme in poultry, wild birds, migratory birds; molecular epidemiology as
well as risk assessment for the virus with pandemic potential are needed in dealing with emergence
of new influenza virus in the country.
3.Molecular Characterization of Avian Infectious Bronchitis Virus Isolated in Malaysia during 2014-2016
Leow, B.L. ; Syamsiah Aini, S ; Faizul Fikri, M.Y ; Muhammad Redzwan, S. ; Khoo, C.K ; Ong, G.H. ; Basirah, M.A ; Norazura, B. ; Mazaitul, Z. ; Mohd Khairil, A. ; Mohd. Jihan, R ; Sohayati, A.R ; Chandrawathani, P.
Tropical Biomedicine 2018;35(4):1092-1106
Avian Infectious Bronchitis (IB) is a highly contagious disease which can cause
huge economic losses to the poultry industry. Forty five IB viruses (IBV) were isolated
from poultry in Malaysia during 2014-2016. Phylogenetic analysis of the spike glycoprotein
1 (S1) gene revealed that all isolates were clustered into five distinct groups. The
predominant type of IBV isolated was QX strains (47%), second was 4/91 type (27%),
followed by Malaysian strain MH5365/95 (13%), Massachusetts type (11%) and finally
Taiwanese strains (2%). Four types of S1 protein cleavage recognition motifs were found
among the isolates which includes HRRRR, RRSRR, RRFRR and RRVRR. To our knowledge,
this is the first report describing the motif RRVRR and are unique to Malaysian strains. Six
IBVs were grouped in Malaysian MH5365/95 strains. Among these, one isolate was different
from others where it only shared 82% identity with MH5365/95 and to others. It formed its
own branch in the Malaysian cluster suggesting it may be a variant unique to Malaysia.
Alignment analysis of the S1 amino acid sequences indicated that point mutations, insertions
and deletions contribute to the divergence of IB variants. This study indicated at least five
groups of IBV are circulating in Malaysia with most of the isolates belonged to QX strains.
As new IBV variants continue to emerge, further study need to be carried out to determine
whether the current available vaccine is able to give protection against the circulating
virus.