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Sequence and phylogenetic analysis of S1, S2, M, and N genes of infectious bronchitis virus isolates from Malaysia

Zulperi ZM, Omar AR, Arshad SS

Virus Genes, 2009 Jun;38(3):383-91.
PMID: 19242786 DOI: 10.1007/s11262-009-0337-2

Two Malaysian infectious bronchitis virus isolates, MH5365/95 and V9/04 were characterized based on sequence and phylogenetic analyses of S1, S2, M, and N genes. Nucleotide sequence alignments revealed many point mutations, short deletions, and insertions in S1 region of both IBV isolates. Phylogenetic analysis of S1 gene and sequences analysis of M gene indicated that MH5365/95 and V9/04 belong to non-Massachusetts strain. However, both isolates share only 77% identity. Analysis based on S1 gene showed that MH5365/95 shared more than 87% identity to several Chinese strains. Meanwhile, V9/04 showed only 67-77% identity to all the previously studied IBV strains included in this study suggesting it is a variant of IBV isolate that is unique to Malaysia. Phylogenetic analysis suggests, although both isolates were isolated 10 years apart from different states in Malaysia, they shared a common origin. Analysis based on S2 and N genes indicated that both strains are highly related to each other, and there are fewer mutations which occurred in the respective genes.
Fulltext Synthesis and characterizations iron oxide carbon nanotubes nanocomposite by laser ablation for anti-microbial applications

Al-Salih M, Samsudin S, Arshad SS

J Genet Eng Biotechnol, 2021 May 18;19(1):76.
PMID: 34003402 DOI: 10.1186/s43141-021-00161-y

BACKGROUND: Environmental contamination by microbes is a major public health concern. A damp environment is one of the potential sources for microbe proliferation. Smart synthesis nanocatalytic coatings on surfaces, food, and material from different pathogen bacteria can inhibit using the Fe3O4/CNTs as anti-microbial growth can effectively curb this growing threat. In this present work, the anti-microbial efficacy of synthesis of a compound nanoparticle-containing iron oxide-multi-walled carbon nanotube was combined by laser ablation PLAL and explored the anti-bacterial action of colloidal solution of Fe3O4/CNTs NPs that was evaluated against bacteria which is classified as gram-negative (Escherichia coli (E. coli), Klebsiella pneumonia (K. pneumonia), and also that is identified as gram-positive (Streptococcus pyogenes (S .pyogenes) and Staphylococcus aureus (S. aureus) under visible light irradiation.
RESULTS: Doping of a minute fraction of iron(III) salt (0.5 mol%) in a volatile solvent (ethanol) was carried out via the sol-gel technique. Fe3O4 was further calcined at various temperatures (in the range of 500-700 °C) to evaluate the thermal stability of the Fe3O4 nanoporous oxidizer nanoparticles. The physicochemical properties of the samples were characterized through X-ray diffraction (XRD), atomic force microscopy (AFM), attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR), and UV-Visible spectroscopy techniques. XRD results revealed that the nanoparticles framework of Fe3O4 was maintained well up to 650 °C by the Fe dopant. UV-Vis results suggested that absorption property of combination Fe3O4/CNTs nanopowder by PLAL was enhanced and the band gap is reduced into 2.0 eV.
CONCLUSIONS: Density functional theory (DFT) studies emphasize the introduction of Fe+ and Fe2+ ions by replacing other ions in the CNT lattice, therefore creating oxygen vacancies. These further promoted anti-microbial efficiency. A significantly high bacterial inactivation that indicates results was evaluated and that the mean estimations of restraint were determined from triple assessment in every appraisal at 400 ml which represent the best anti-bacterial action against gram-positive and gram-negative microbes.
Fulltext Isolation and Metagenomic Identification of Avian Leukosis Virus Associated with Mortality in Broiler Chicken

Bande F, Arshad SS, Omar AR

Adv Virol, 2016;2016:9058403.
PMID: 27597865 DOI: 10.1155/2016/9058403

Avian leukosis virus (ALV) belongs to the family Retroviridae and causes considerable economic losses to the poultry industry. Following an outbreak associated with high mortality in a broiler flock in northern part of Malaysia, kidney tissues from affected chickens were submitted for virus isolation and identification in chicken embryonated egg and MDCK cells. Evidence of virus growth was indicated by haemorrhage and embryo mortality in egg culture. While viral growth in cell culture was evidenced by the development of cytopathic effects. The isolated virus was purified by sucrose gradient and identified using negative staining transmission electron microscopy. Further confirmation was achieved through next-generation sequencing and nucleotide sequence homology search. Analysis of the viral sequences using the NCBI BLAST tool revealed 99-100% sequence homology with exogenous ALV viral envelope protein. Phylogenetic analysis based on partial envelope sequences showed the Malaysian isolate clustered with Taiwanese and Japanese ALV strains, which were closer to ALV subgroup J, ALV subgroup E, and recombinant A/E isolates. Based on these findings, ALV was concluded to be associated with the present outbreak. It was recommended that further studies should be conducted on the molecular epidemiology and pathogenicity of the identified virus isolate.
Fulltext Molecular detection, phylogenetic analysis, and identification of transcription motifs in feline leukemia virus from naturally infected cats in malaysia

Bande F, Arshad SS, Hassan L, Zakaria Z

Vet Med Int, 2014;2014:760961.
PMID: 25506469 DOI: 10.1155/2014/760961

A nested PCR assay was used to determine the viral RNA and proviral DNA status of naturally infected cats. Selected samples that were FeLV-positive by PCR were subjected to sequencing, phylogenetic analysis, and motifs search. Of the 39 samples that were positive for FeLV p27 antigen, 87.2% (34/39) were confirmed positive with nested PCR. FeLV proviral DNA was detected in 38 (97.3%) of p27-antigen negative samples. Malaysian FeLV isolates are found to be highly similar with a homology of 91% to 100%. Phylogenetic analysis revealed that Malaysian FeLV isolates divided into two clusters, with a majority (86.2%) sharing similarity with FeLV-K01803 and fewer isolates (13.8%) with FeLV-GM1 strain. Different enhancer motifs including NF-GMa, Krox-20/WT1I-del2, BAF1, AP-2, TBP, TFIIF-beta, TRF, and TFIID are found to occur either in single, duplicate, triplicate, or sets of 5 in different positions within the U3-LTR-gag region. The present result confirms the occurrence of FeLV viral RNA and provirus DNA in naturally infected cats. Malaysian FeLV isolates are highly similar, and a majority of them are closely related to a UK isolate. This study provides the first molecular based information on FeLV in Malaysia. Additionally, different enhancer motifs likely associated with FeLV related pathogenesis have been identified.
Detection of avian leukosis virus subgroup J in chicken flocks from Malaysia and their molecular characterization

Thapa BR, Omar AR, Arshad SS, Hair-Bejo M

Avian Pathol, 2004 Jun;33(3):359-63.
PMID: 15223566

Previously we have shown that avian leukosis virus subgroup J (ALV-J) might be present in chicken flocks from Malaysia based on serological study and also on detection of tissue samples with myelocytic infiltration. In this study, the polymerase chain reaction was used to detect ALV-J sequences from archived frozen samples. Out of 21 tissue samples examined, 16 samples were positive for proviral DNA and four samples for ALV-J RNA. However, only nine samples were found positive for myelocytic infiltration. A total of 465 base pairs equivalent to positions 5305 to 5769 of HPRS-103 from each of the viral RNA positive samples were characterized. Sequence analysis indicated that the samples showed high identity (95.9 to 98.2%) and were close to HPRS-103 with identities between 97.4 and 99.3%. This study indicates that ALV-J-specific sequences can be detected by polymerase chain reaction from frozen tissue samples with and without myelocytic infiltration.
Fulltext Molecular Detection, Risk Factors and Public Awareness of Avian Bornavirus among Captive and Non-captive Birds in Peninsular Malaysia

Lutpi SM, Abu J, Arshad SS, Rahaman NY

J Vet Res, 2022 Dec;66(4):523-535.
PMID: 36846031 DOI: 10.2478/jvetres-2022-0061

INTRODUCTION: Proventricular dilatation disease (PDD) is caused by avian bornavirus (ABV) has been identified in psittacine, non-psittacine birds and waterfowl. Birds may show signs of gastrointestinal tract deficit or neurological dysfunction or even both. The objectives of this study were to determine the molecular prevalence, risk factors and public awareness of ABV and PDD among captive and non-captive birds in Peninsular Malaysia.
MATERIAL AND METHODS: A total of 344 cloacal swabs or faeces were collected and subjected to detection using the RT-PCR assay. Meanwhile, KAP questionnaires were distributed by using the Google forms platform.
RESULTS: Molecular prevalence studies revealed that 4.5% (9/201) of the pet birds were ABV-positive, whereas 0% (0/143) in waterfowl. Nine positive pet birds were identified to be PaBV-2, which is closest to ABV isolates EU781967 (USA). Among the risk factors analysed, category, age and, location, were found to show an association with the ABV positivity. The KAP survey result showed: the respondents have low knowledge (32.9%), however, they showed positive attitude (60.8%) and good practice (94.9%). The association between knowledge, attitude and practice showed that there was a significant association between knowledge-attitude and also attitude-practice (P<0.05).
CONCLUSION: This study proved that avian bornavirus (ABV) causes proventricular dilatation disease (PDD) among a group of pet birds of Psittaciformes, but it is present in Peninsular Malaysia with a low prevalence rate. Furthermore, in addition to the useful databases obtained from this study, the level of public awareness on the importance of avian bornavirus that causes fatal disorders among a wide range of bird species is satisfactorily raised.
Performance of an RT-nested PCR ELISA for detection of Newcastle disease virus

Kho CL, Mohd-Azmi ML, Arshad SS, Yusoff K

J Virol Methods, 2000 Apr;86(1):71-83.
PMID: 10713378

A sensitive and specific RT-nested PCR coupled with an ELISA detection system for detecting Newcastle disease virus is described. Two nested pairs of primer which were highly specific to all the three different pathotypes of NDV were designed from the consensus fusion gene sequence. No cross-reactions with other avian infectious agents such as infectious bronchitis virus, infectious bursal disease virus, influenza virus, and fowl pox virus were observed. Based on agarose electrophoresis detection, the RT-nested PCR was about 100 times more sensitive compared to that of a non-nested RT-PCR. To facilitate the detection of the PCR product, an ELISA detection method was then developed to detect the amplified PCR products and it was shown to be ten times more sensitive than gel electrophoresis. The efficacy of the nested PCR-ELISA was also compared with the conventional NDV detection method (HA test) and non-nested RT-PCR by testing against a total of 35 tissue specimens collected from ND-symptomatic chickens. The RT-nested PCR ELISA found NDV positive in 21 (60%) tissue specimens, while only eight (22.9%) and two (5.7%) out of 35 tissue specimens were tested NDV positive by both the non-nested RT-PCR and conventional HA test, respectively. Due to its high sensitivity for the detection of NDV from tissue specimens, this PCR-ELISA based diagnostic test may be useful for screening large number of samples.
Fulltext Peripheral blood gene expression profile of atherosclerotic coronary artery disease in patients of different ethnicity in Malaysia

Abdullah MH, Othman Z, Noor HM, Arshad SS, Yusof AK, Jamal R, et al.

J Cardiol, 2012 Sep;60(3):192-203.
PMID: 22738689 DOI: 10.1016/j.jjcc.2012.05.009

The molecular basis of coronary artery disease (CAD) has been widely studied in the western world but there is no published work on the Malaysian population. This study looked at the global gene expression profiling of the peripheral blood of patients with CAD from the 3 main ethnic groups in Malaysia. Male subjects selected were based on angiographically confirmed CAD (≥50% stenosis) and normal control subjects (0% stenosis) with age range of 55.6±5.3 and 51.0±5.5 years, respectively. The global gene expression of 12 angiographically documented CAD patients and 11 matched control subjects were performed. The combined group samples identified 6 up regulated differential expression (DE) genes (GHRL, LTA, CBS, HP, ITGA2B, and OLR1) and 12 down regulated DE genes (IL18R1, ITGA2B, IL18RAP, HP, OLR1, SOD2 ITGB3, IL1B, MMP9, PLA2G7, UTS2, and CBS) to be involved in CAD at the fold change of 1.3 with fault discovery rate (FDR) of 1%. Three genes, MMP9, IL1B, and SOD2 were down regulated in all the 3 ethnic groups making them potential biomarker candidates for CAD across all three ethnicities. Further verification in a cohort study is needed.
Fulltext Diagnostic methods for feline coronavirus: a review

Sharif S, Arshad SS, Hair-Bejo M, Omar AR, Zeenathul NA, Alazawy A

Vet Med Int, 2010;2010.
PMID: 20798771 DOI: 10.4061/2010/809480

Feline coronaviruses (FCoVs) are found throughout the world. Infection with FCoV can result in a diverse range of signs from clinically inapparent infections to a highly fatal disease called feline infectious peritonitis (FIP). FIP is one of the most serious viral diseases of cats. While there is neither an effective vaccine, nor a curative treatment for FIP, a diagnostic protocol for FCoV would greatly assist in the management and control of the virus. Clinical findings in FIP are non-specific and not helpful in making a differential diagnosis. Haematological and biochemical abnormalities in FIP cases are also non-specific. The currently available serological tests have low specificity and sensitivity for detection of active infection and cross-react with FCoV strains of low pathogenicity, the feline enteric coronaviruses (FECV). Reverse transcriptase polymerase chain reaction (RT-PCR) has been used to detect FCoV and is rapid and sensitive, but results must be interpreted in the context of clinical findings. At present, a definitive diagnosis of FIP can be established only by histopathological examination of biopsies. This paper describes and compares diagnostic methods for FCoVs and includes a brief account of the virus biology, epidemiology, and pathogenesis.
Prevalence of feline coronavirus in two cat populations in Malaysia

Sharif S, Arshad SS, Hair-Bejo M, Omar AR, Zeenathul NA, Hafidz MA

J Feline Med Surg, 2009 Dec;11(12):1031-4.
PMID: 19818660 DOI: 10.1016/j.jfms.2009.08.005

The prevalence of feline coronavirus (FCoV) was studied in two catteries in Malaysia. Rectal swabs or faecal samples were collected from a total of 44 clinically healthy Persian purebred and mix-breed cats. RNA extracted from the faecal material was subjected to a reverse transcription-polymerase chain reaction (RT-PCR) using primers flanking for a conserved region of the virus genome. The overall prevalence of FCoV infection was 84% and the infection rate was higher in Persian purebred cats (96%) than mix-breed cats (70%). There was no significant association between the age or gender of tested cats and shedding the virus. This study is the first PCR-based survey for FCoV in Malaysia and showed the ubiquitous presence of FCoV in Malaysian cat colonies.
Fulltext Pathogenesis and Diagnostic Approaches of Avian Infectious Bronchitis

Bande F, Arshad SS, Omar AR, Bejo MH, Abubakar MS, Abba Y

Adv Virol, 2016;2016:4621659.
PMID: 26955391 DOI: 10.1155/2016/4621659

Infectious bronchitis (IB) is one of the major economically important poultry diseases distributed worldwide. It is caused by infectious bronchitis virus (IBV) and affects both galliform and nongalliform birds. Its economic impact includes decreased egg production and poor egg quality in layers, stunted growth, poor carcass weight, and mortality in broiler chickens. Although primarily affecting the respiratory tract, IBV demonstrates a wide range of tissues tropism, including the renal and reproductive systems. Thus, disease outcome may be influenced by the organ or tissue involved as well as pathotypes or strain of the infecting virus. Knowledge on the epidemiology of the prevalent IBV strains in a particular region is therefore important to guide control and preventions. Meanwhile previous diagnostic methods such as serology and virus isolations are less sensitive and time consuming, respectively; current methods, such as reverse transcription polymerase chain reaction (RT-PCR), Restriction Fragment Length Polymorphism (RFLP), and sequencing, offer highly sensitive, rapid, and accurate diagnostic results, thus enabling the genotyping of new viral strains within the shortest possible time. This review discusses aspects on pathogenesis and diagnostic methods for IBV infection.
Fulltext Scoring System for Lesions Induced by Different Strains of Newcastle Disease Virus in Chicken

Hussein EA, Hair-Bejo M, Adamu L, Omar AR, Arshad SS, Awad EA, et al.

Vet Med Int, 2018;2018:9296520.
PMID: 30631413 DOI: 10.1155/2018/9296520

Newcastle disease virus strains are velogenic, mesogenic, and lentogenic. This study aims to design a scoring system for lesions induced by different strains of Newcastle disease virus in chicken. Three experiments were conducted. In experiments 1 and 2, chickens were divided into infected and control groups. Infected groups of experiments 1 and 2 consisted of 6 and 24 specific pathogen-free (SPF) chickens, respectively. Control groups in experiments 1 and 2 consisted of 6 and 15 SPF chickens, respectively. In infected groups, infection was induced by intranasal administration of 105 50% EID50/0.1 mL of velogenic Newcastle disease virus strain (vNDV). Infected chickens in experiment 1 were euthanised by cervical dislocation on days 3, 6, and 7 postinoculation (pi). Infected chickens in experiment 2 were euthanised at hours (hrs) 2, 4, 6, 12 and days 1, 2, 4, and 6 pi. Chickens of the control group in experiment 1 were euthanised on days 3 and 7 pi, whereas control group chickens in experiment 2 were euthanised on days 0, 1, 2, 4, and 6 pi. Then in experiment 3, 15 SPF chickens were divided into three groups; in the first group, 5 SPF chickens were infected with vNDV, in the second group, 5 SPF chickens were infected with lentogenic NDV (lNDV) (103.0 EID50/0.1 mL), and the third group was kept without infection as a control group. Chickens were euthanised on day 5 pi. In all previous experiments, tissues of brain, trachea, lung, caecal tonsil, liver, kidney, spleen, heart, proventriculus, intestine, and thymus were collected, fixed in 10% buffered formalin, embedded in paraffin, and sectioned. HS staining was applied. Tissues were examined under light microscope and changes were recorded. A scoring system was designed for lesions induced by different strains of NDV and, accordingly, lesions were scored. The scoring system was found helpful in the evaluation of disease severity.
Development and partial characterization of new marine cell line from brain of Asian sea bass Lates calcarifer for virus isolation

Hasoon MF, Daud HM, Abdullah AA, Arshad SS, Bejo HM

In Vitro Cell Dev Biol Anim, 2011 Jan;47(1):16-25.
PMID: 21082288 DOI: 10.1007/s11626-010-9348-5

A new cell line, Asian sea bass brain (ASBB), was derived from the brain tissue of Asian sea bass Lates calcarifer. This cell line was maintained in Leibovitz L-15 media supplemented with 10% fetal bovine serum (FBS). The ASBB cell line was subcultured more than 60 times over a period of 15 mo. The ASBB cell line consists predominantly of fibroblastic-like cells and was able to grow at temperatures between 20°C and 30°C with an optimum temperature of 25°C. The growth rate of these cells increased as the proportion of FBS increased from 2% to 20% at 25°C with optimum growth at the concentrations of 10% or 15% FBS. Polymerase chain reaction products were obtained from ASBB cells and tissues of sea bass with primer sets of microsatellite markers of sea bass. An isolate of piscine nodavirus from juveniles of marine fish species tested positive by IQ2000 kit for viral nervous necrosis detection and was examined for its infectivity to a fish cell line of ASBB. A marine fish betanodavirus was tested to determine the susceptibility of this new cell line in comparison with commercial highly permissive SSN-1 cells. The ASBB cell line was found to be susceptible to nodavirus (RGNNV genotype), and the infection was confirmed by comparison cytopathic effect (CPE) with commercial SSN-1 and reverse transcriptase-polymerase chain reaction. A nodavirus was further elucidated by electron microscopy, and the virus tested was shown to induce CPE on ASBB cells with significant high titer. This suggests that the ASBB cell line has good potential for the isolation of fish viruses.
Fulltext Serological evidence of West Nile viral infection in archived swine serum samples from Peninsular Malaysia

Mohammed MN, Yasmin AR, Noraniza MA, Ramanoon SZ, Arshad SS, Bande F, et al.

J Vet Sci, 2021 May;22(3):e29.
PMID: 33908203 DOI: 10.4142/jvs.2021.22.e29

West Nile virus (WNV), a neurotropic arbovirus, has been detected in mosquitos, birds, wildlife, horses, and humans in Malaysia, but limited information is available on WNV infection in Malaysian pigs. We tested 80 archived swine serum samples for the presence of WNV antibody and West Nile (WN) viral RNA using ID Screen West Nile Competition Multi-species enzyme-linked immunosorbent assay kits and WNV-specific primers in reverse transcription polymerase chain reaction assays, respectively. A WNV seroprevalence of 62.5% (50/80) at 95% confidence interval (51.6%-72.3%) was recorded, with a significantly higher seroprevalence among young pigs (weaner and grower) and pigs from south Malaysia. One sample was positive for Japanese encephalitis virus antibodies; WN viral RNA was not detected in any of the serum samples.
Fulltext Prediction and In Silico Identification of Novel B-Cells and T-Cells Epitopes in the S1-Spike Glycoprotein of M41 and CR88 (793/B) Infectious Bronchitis Virus Serotypes for Application in Peptide Vaccines

Bande F, Arshad SS, Hair Bejo M, Kadkhodaei S, Omar AR

Adv Bioinformatics, 2016;2016:5484972.
PMID: 27667997 DOI: 10.1155/2016/5484972

Bioinformatic analysis was used to predict antigenic B-cell and T-cell epitopes within the S1 glycoprotein of M41 and CR88 IBV strains. A conserved linear B-cell epitope peptide, YTSNETTDVTS(175-185), was identified in M41 IBV strains while three such epitopes types namely, VSNASPNSGGVD(279-290), HPKCNFRPENI(328-338), and NETNNAGSVSDCTAGT(54-69), were predicted in CR88 IBV strains. Analysis of MHCI binding peptides in M41 IBV strains revealed the presence of 15 antigenic peptides out of which 12 were highly conserved in 96-100% of the total M41 strains analysed. Interestingly three of these peptides, GGPITYKVM(208), WFNSLSVSI(356), and YLADAGLAI(472), relatively had high antigenicity index (>1.0). On the other hand, 11 MHCI binding epitope peptides were identified in CR88 IBV strains. Of these, five peptides were found to be highly conserved with a range between 90% and 97%. However, WFNSLSVSL(358), SYNISAASV(88), and YNISAASVA(89) peptides comparably showed high antigenicity scores (>1.0). Combination of antigenic B-cells and T-cells peptides that are conserved across many strains as approach to evoke humoral and CTL immune response will potentially lead to a broad-based vaccine that could reduce the challenges in using live attenuated vaccine technology in the control of IBV infection in poultry.
Fulltext Propagation and Molecular Characterization of Bioreactor Adapted Very Virulent Infectious Bursal Disease Virus Isolates of Malaysia

Lawal N, Hair-Bejo M, Arshad SS, Omar AR, Ideris A

J Pathog, 2018;2018:1068758.
PMID: 30245887 DOI: 10.1155/2018/1068758

Two Malaysian very virulent infectious bursal disease virus (vvIBDV) strains UPM0081 (also known as B00/81) and UPM190 (also known as UPM04/190) isolated from local IBD outbreaks in 2000 and 2004, respectively, were separately passaged for 12 consecutive times in 11-day-old specific pathogen free (SPF) chicken embryonated eggs (CEE) via the chorioallantoic membrane (CAM) route. The CEE passage 8 (EP8) isolates were passaged once in BGM-70 cell line yielding UPM0081EP8BGMP1 and UPM190EP8BGMP1, while the EP12 isolates were passaged 15 times in BGM-70 cell line yielding UPM0081EP12BGMP15 and UPM190EP12BGMP15 using T25 tissue culture flask. These isolates were all propagated once in bioreactor using cytodex 1 as microcarrier at 3 g per liter (3 g/L) yielding UPM0081EP8BGMP1BP1, UPM190EP8BGMP1BP1, UPM0081EP12BGMP15BP1, and UPM190EP12BGMP15BP1 isolates. The viruses were harvested at 3 days after inoculation, following the appearance of cytopathic effects (CPE) characterized by detachment from the microcarrier using standard protocol and filtered using 0.2 μm syringe filter. The filtrates were positive for IBDV by RT-PCR and immunofluorescence. Sequence and phylogenetic tree analysis indicated that the isolates were of the vvIBDV strains and were not different from the flask propagated parental viruses.
Fulltext Immunological Analysis of Nodavirus Capsid Displaying the Domain III of Japanese Encephalitis Virus Envelope Protein

Kumar K, Ong HK, Tan WS, Arshad SS, Ho KL

Pharmaceutics, 2021 Nov 01;13(11).
PMID: 34834244 DOI: 10.3390/pharmaceutics13111826

Japanese encephalitis virus (JEV) is the pathogen that causes Japanese encephalitis (JE) in humans and horses. Lethality of the virus was reported to be between 20-30%, of which, 30-50% of the JE survivors develop neurological and psychiatric sequelae. Attributed to the low effectiveness of current therapeutic approaches against JEV, vaccination remains the only effective approach to prevent the viral infection. Currently, live-attenuated and chimeric-live vaccines are widely used worldwide but these vaccines pose a risk of virulence restoration. Therefore, continuing development of JE vaccines with higher safety profiles and better protective efficacies is urgently needed. In this study, the Macrobrachium rosenbergii nodavirus (MrNV) capsid protein (CP) fused with the domain III of JEV envelope protein (JEV-DIII) was produced in Escherichia coli. The fusion protein (MrNV-CPJEV-DIII) assembled into virus-like particles (VLPs) with a diameter of approximately 18 nm. The BALB/c mice injected with the VLPs alone or in the presence of alum successfully elicited the production of anti-JEV-DIII antibody, with titers significantly higher than that in mice immunized with IMOJEV, a commercially available vaccine. Immunophenotyping showed that the MrNV-CPJEV-DIII supplemented with alum triggered proliferation of cytotoxic T-lymphocytes, macrophages, and natural killer (NK) cells. Additionally, cytokine profiles of the immunized mice revealed activities of cytotoxic T-lymphocytes, macrophages, and NK cells, indicating the activation of adaptive cellular and innate immune responses mediated by MrNV-CPJEV-DIII VLPs. Induction of innate, humoral, and cellular immune responses by the MrNV-CPJEV-DIII VLPs suggest that the chimeric protein is a promising JEV vaccine candidate.
First molecular detection of porcine bocavirus in Malaysia

Mohan Jacob D, Lee CY, Arshad SS, Selvarajah GT, Bande F, Ong BL, et al.

Trop Anim Health Prod, 2018 Apr;50(4):733-739.
PMID: 29243138 DOI: 10.1007/s11250-017-1489-z

Several strains of porcine bocaviruses have been reported worldwide since their first detection in Sweden in 2009. Subsequently, the virus has been reported to be associated with gastrointestinal and respiratory signs in weaner and grower pigs. Although Malaysia is host to a self-sufficient swine livestock industry, there is no study that describes porcine bocavirus in the country. This report is the first to describe porcine bocavirus (PBoV) in Malaysian swine herds. PBoV was identified in various tissues from sick and runt pigs using the conventional PCR method with primers targeting conserved regions encoding for the nonstructural protein (NS1) gene. Out of 103 samples tested from 17 pigs, 32 samples from 15 pigs were positive for porcine bocavirus. In addition, a higher detection rate was identified from mesenteric lymph nodes (52.9%), followed by tonsil (37.0%), and lungs (33.3%). Pairwise comparison and phylogenetic analyses based on a 658-bp fragment of NS1 gene revealed that the Malaysian PBoV strains are highly similar to PBoV3 isolated in Minnesota, USA. The presence of porcine bocavirus in Malaysia and their phylogenetic bond was marked for the first time by this study. Further studies will establish the molecular epidemiology of PBoV in Malaysia and clarify pathogenicity of the local isolates.
Fulltext Adaptation and Molecular Characterization of Two Malaysian Very Virulent Infectious Bursal Disease Virus Isolates Adapted in BGM-70 Cell Line

Lawal N, Hair-Bejo M, Arshad SS, Omar AR, Ideris A

Adv Virol, 2017;2017:8359047.
PMID: 29230245 DOI: 10.1155/2017/8359047

Two Malaysian very virulent infectious bursal disease virus (vvIBDV) strains UPM0081 and UPM190 (also known as UPMB00/81 and UPM04/190, respectively) isolated from local IBD outbreaks were serially passaged 12 times (EP12) in specific pathogen free (SPF) chicken embryonated eggs (CEE) by chorioallantoic membrane (CAM) route. The EP12 isolate was further adapted and serially propagated in BGM-70 cell line up to 20 passages (P20). Characteristic cytopathic effects (CPEs) were subtly observed at P1 in both isolates 72 hours postinoculation (pi). The CPE became prominent at P5 with cell rounding, cytoplasmic vacuoles, granulation, and detachment from flask starting from day 3 pi, up to 7 days pi with titers of 109.50 TCID50/mL and log109.80 TCID50/mL for UPM0081 and UPM190, respectively. The CPE became subtle at P17 and disappeared by P18 and P19 for UPM0081 and UPM190, respectively. However, the presence of IBDV was confirmed by immunoperoxidase, immunofluorescence, and RT-PCR techniques. Phylogenetic analysis showed that these two isolates were of the vvIBDV. It appears that a single mutation of UPM190 and UPM0081 IBDV isolates at D279N could facilitate vvIBDV strain adaptability in CEE and BGM-70 cultures.
Global distributions and strain diversity of avian infectious bronchitis virus: a review

Bande F, Arshad SS, Omar AR, Hair-Bejo M, Mahmuda A, Nair V

Anim Health Res Rev, 2017 Jun;18(1):70-83.
PMID: 28776490 DOI: 10.1017/S1466252317000044

The poultry industry faces challenge amidst global food security crisis. Infectious bronchitis is one of the most important viral infections that cause huge economic loss to the poultry industry worldwide. The causative agent, infectious bronchitis virus (IBV) is an RNA virus with great ability for mutation and recombination; thus, capable of generating new virus strains that are difficult to control. There are many IBV strains found worldwide, including the Massachusetts, 4/91, D274, and QX-like strains that can be grouped under the classic or variant serotypes. Currently, information on the epidemiology, strain diversity, and global distribution of IBV has not been comprehensively reported. This review is an update of current knowledge on the distribution, genetic relationship, and diversity of the IBV strains found worldwide.

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