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Isolation, identification and characterization of chicken anaemia virus in Malaysia

Chowdhury SM, Omar AR, Aini I, Hair-Bejo M, Jamaluddin AA, Kono Y, et al.

J. Biochem. Mol. Biol. Biophys., 2002 Aug;6(4):249-55.
PMID: 12186740

A study was conducted to isolate and identify chicken anaemia virus (CAV) from field samples of clinically infected broiler chickens in Malaysia. A total of 125 samples were collected from chickens aged 2-6 weeks with clinically depressed and retarded growth, of which five samples were found positive to CAV directly by polymerase chain reaction (PCR). Later, five isolates of CAV from the respective five PCR positive samples were isolated in MDCC-MSB1 cells at passage 4 based on cytopathic effects, PCR and indirect immunofluorescent antibody test. The isolates were identified as BL-1, BL-2, BL-3, BL-4 and BL-5. These CAV isolates were found to resist treatment with chloroform and heat at 37 degrees C for 2 h, 56 degrees C for 30 min and 70 degrees C for 5 min. One of the isolates, BL-5 produced significant reduction (p < 0.001) of hematocrit values (9-19%), pale bone marrow, thymus atrophy and haemorrhages in skin/muscle when inoculated into 1-day old SPF chickens. Restriction enzyme digestion of 926 bp genomic fragments of all the isolates including Cux-1 isolate with HindIII exhibited a similar pattern of bands in 2% agarose gel. The present findings confirmed the presence of CAV in Malaysia.
Characterization and identification of Oya virus, a Simbu serogroup virus of the genus Bunyavirus, isolated from a pig suspected of Nipah virus infection

Kono Y, Yusnita Y, Mohd Ali AR, Maizan M, Sharifah SH, Fauzia O, et al.

Arch Virol, 2002 Aug;147(8):1623-30.
PMID: 12181680

A virus, named Oya virus, was isolated in Vero cell cultures from the lungs of a pig suspected of Nipah virus infection. The virus was revealed as a spherical enveloped RNA virus with a diameter of 79 nm. For identification of Oya virus, RT-PCR was performed. A common primer set for S-RNA of the Simbu serogroup of the genus Bunyavirus was able to amplify a cDNA from Oya virus RNA. The sequence data of the product revealed that the partial gene of Oya virus S-RNA segment had 65-70% homology with published cDNA sequences of Simbu serogroup viruses. The phylogenetic analysis of the data showed that the Oya virus is grouped in Simbu serogroup, but is genetically distinct from the serogroup viruses that have been analyzed molecularly. Serological surveys revealed that the virus distributed widely and densely in Malaysia.
Development of a modified method for sequencing high G:C regions in chicken anemia virus genome

Chowdhury SM, Omar AR, Aini I, Hair-Bejo M, Jamaluddin AA, Wan KL, et al.

J. Biochem. Mol. Biol. Biophys., 2002 Jun;6(3):229-32.
PMID: 12186760

Two areas in the chicken anemia virus (CAV) genome have high G:C content with secondary structures. These two G:C rich areas could not be sequenced with Perkin Elmer's Big Dye Terminator Cycle Sequencing Kit. Several modifications were carried out to solve the problem. Finally, a package of modified method was developed to sequence the high G:C areas. The result showed that the Perkin Elmer's Big Dye Terminator Cycle Sequencing Kit with the normal procedures are not suitable for sequencing the high G:C regions of the CAV genome. The present developed method made the Perkin Elmer's Kit useful for the first time to sequence the G:C rich hairpin structures of the CAV genome. The system may be useful to sequence all other G:C rich DNA templates.
Pathogenicity, sequence and phylogenetic analysis of Malaysian Chicken anaemia virus obtained after low and high passages in MSB-1 cells

Chowdhury SM, Omar AR, Aini I, Hair-Bejo M, Jamaluddin AA, Md-Zain BM, et al.

Arch Virol, 2003 Dec;148(12):2437-48.
PMID: 14648297

Specific-pathogen-free (SPF) chickens inoculated with low passage Chicken anaemia virus (CAV), SMSC-1 and 3-1 isolates produced lesions suggestive of CAV infection. Repeated passages of the isolates in cell culture until passage 60 (P60) and passage 123 produced viruses that showed a significantly reduced level of pathogenicity in SPF chickens compared to the low passage isolates. Sequence comparison indicated that nucleotide changes in only the coding region of the P60 passage isolates were thought to contribute to virus attenuation. Phylogenetic analysis indicated that SMSC-1 and 3-1 were highly divergent, but their P60 passage derivatives shared significant homology to a Japanese isolate A2.
Fulltext Two new species of Begonia, B. moneta and B. peridoticola (Begoniaceae) from Sabah, Malaysia

Peng CI, Lin CW, Repin R, Kono Y, Leong WC, Chung KF

Bot Stud, 2015 Dec;56(1):7.
PMID: 28510816 DOI: 10.1186/s40529-015-0087-5

BACKGROUND: Mount Kinabalu, reknowned for its high biodiversity and endemism, is a National Park in the State of Sabah on the northern end of the island of Borneo. Every year many visit the higher part of the Kinabalu National Park, while most lowland forests in the Park are under-explored. Two unknown species of Begonia were collected from a peridotic (ultramafic) cliff in the Kinabalu National Park at ca. 400 m elevation.
RESULTS: The two species are named B. moneta C.-I Peng, Rimi & C. W. Lin and B. peridoticola Rimi, C.-I Peng & C. W. Lin. Begonia moneta (sect. Baryandra) is similar to B. gueritziana Gibbs, a widespread species of the same section in Borneo, differing in the peltate (vs. basifixed) leaves and the smaller flower parts. Also, their chromosome numbers are different (B. moneta, 2n = 30; B. gueritziana, 2n = 28). The peltate and succulent foliage of B. moneta is also reminiscent of B. burttii Kiew & S. Julia and B. payung S. Julia & Kiew, both of sect. Reichenheimia, from Sarawak. Begonia moneta is distinct from the two species in having branched (vs. entire) placental lamellae. Additionally, B. moneta differs from B. burttii in having 4 (vs. 5) tepals in pistillate flowers and markedly unequal (vs. equal) fruit wings. Begonia moneta differs from B. payung in the smaller leaves and conspicuously winged (vs. wingless) capsules. Begonia peridoticola (sect. Petermannia) resembles B. punchak Kiew & S. Julia from limestone areas in Kuching Division, Sarawak, differing in the entire leaf margin (vs. distantly dentate), much larger capsular wings (8-11 mm vs. 2-3 mm wide) and yellow, spiral (vs. crimson, U-shaped) styles.
CONCLUSION: A careful study of the herbarium materials and literature supports the recognition of the two new species. Detailed descriptions, line drawings, color plates, chromsome data, foliar SEM observations and comparisons with phenetically similar species are provided to aid in identification.
Encephalitis and retarded growth of chicks caused by Sitiawan virus, a new isolate belonging to the genus Flavivirus

Kono Y, Tsukamoto K, Abd Hamid M, Darus A, Lian TC, Sam LS, et al.

Am J Trop Med Hyg, 2001 5 19;63(1-2):94-101.
PMID: 11358004

A new virus named Sitiawan virus (SV) was isolated from sick broiler chicks in chicken embryos. The virus replicated well with cytopathogenic effect (CPE) in the chicken B-lymphocyte cell line LSCC-BK3. The virus was an enveloped RNA virus of approximately 41 nm in size with hemagglutinating activity (HA) to goose erythrocytes. It was cross-reactive with Japanese encephalitis virus (JEV), a member of flaviviruses by HA inhibition tests but not by cross-virus neutralization tests. The cDNA fragment of NS5 gene was amplified with primers corresponding to NS5 gene of flaviviruses. The nucleotide sequences were 92% homologous to Tembusu virus, a member of the mosquito-borne virus cluster of the genus Flavivirus. In cross-neutralization tests with Tembusu virus, antiserum to SV did not neutralize Tembusu virus, and antiserum to Tembusu virus neutralized more weakly to SV than against homologous virus. These results indicate that SV is a new virus which can be differentiated serologically from Tembusu virus but is otherwise similar with respect to nucleotide sequence. The virus causes encephalitis, growth retardation, and increased blood glucose levels in inoculated chicks.