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  1. Fulltext Serological evidence and experimental infection of cynomolgus macaques with pteropine orthoreovirus reveal monkeys as potential hosts for transmission to humans
    Tan CW, Wittwer K, Lim XF, Uehara A, Mani S, Wang LF, et al.
    Emerg Microbes Infect, 2019;8(1):787-795.
    PMID: 31132935 DOI: 10.1080/22221751.2019.1621668
    Pteropine orthoreoviruses (PRV) are emerging bat-borne viruses with proven zoonotic transmission. We recently demonstrated human exposure to PRV in Singapore, which together with previous reports from Malaysia and Vietnam suggest that human infection of PRV may occur periodically in the region. This raises the question whether bats are the only sources of human infection. In this study, we screened 517 cynomolgus macaques caught in Singapore for evidence of exposure to PRV3M (also known as Melaka virus), which was first isolated from human patients in Melaka, Malaysia. We found that 67 serum samples were PRV3M positive by ELISA and 34 were also positive by virus neutralization assay. To investigate whether monkeys could act as hosts for PRV transmission, we experimentally infected cynomolgus macaques with PRV3M and housed these animals with uninfected monkeys. Although no clinical signs of infection were observed in infected animals, viral RNA was detected in nasal and rectal swabs and all infected macaques seroconverted. Additionally, one of the uninfected animals seroconverted, implying active shedding and transmission of PRV3M. We provide evidence that PRV exposure in the macaque population in Singapore occurs at a relatively high prevalence and this study suggests that cynomolgus macaques may be an intermediate or reservoir host for PRVs.
    Matched MeSH terms: DNA Viruses
  2. Selective inhibition of genes in methicillin resistant Staphylococcus aureus (MRSA) treated with Melastoma malabathricum methanol extract
    Zulaikah Mohamed, Nazlina Ibrahim, Ismail Ahmad
    Sains Malaysiana, 2008;37(1):107-113.
    Methanol extract of Melastoma malabathricum leaves inhibited the growth of Staphylococcus aureus and six clinical isolates of Methicilin Resistant Stapyhlococcus aureus (MRSA 1-6). The minimum inhibitory concentration (MIC) of test substance was 1.565mg/ml and the minimum bacteriocidal concentration (MBC) was 3.125 mg/ml. The methanol extract suppressed RNA synthesis at 10 mg/ml as shown by RNA profile which was devoid of three bands compared to the control. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis using seven primer pairs was only successful in amplifying four cDNA amplicons. The failure to amplify three cDNA amplicons for three primer pairs corresponding to gyrA, femA and nuc genes, implied the possibility of suppression of the corresponding mRNA. Electrophoretic separation of endogenous and exogenuos bacterial proteins showed that three and five protein, respectively were not expressed. One endogenous and three exogenous proteins were over-expressed in treated MRSA compared with untreated control. The results of the molecular and proteomic analyses are in agreement, and based on primers being used, methanol extract of M. malabathricum leaves possibly inhibits MRSA growth through inhibition of DNA synthesis, peptidoglycan production, and nuclease production.
    Keywords: Methicillin resistant Staphylococcus aureus; Melastoma malabathricum; gene expression; protein production
    Matched MeSH terms: DNA, Complementary
  3. Comparison between Pork and wild Boar meat (Sus Scrofa ) by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP)
    Sahilah Abu Mutalib, Wan Sakeenah Wan Nazari, Safiyyah Shahimi, Norhayati Yaakob, Norrakiah Abdullah Sani, Aminah Abdullah, et al.
    Sains Malaysiana, 2012;41:199-204.
    A method of PCR-restriction fragment length polymorphism (RFLP) has been utilized to differentiate the mitochondrial genes of pork and wild boar meat (Sus scrofa). The amplification PCR products of 359 bp and 531 bp were successfully amplified from the cyt b gene of these two meats. The amplification product of pork and wild boar using mt-12S rRNA gene successfully produced a single band with molecular size of 456 bp. Three restriction endonucleases (AluI, HindIII and BsaJI) were used to restrict the amplification products of the mitochondrial genes. The restriction enzymes of AluI and BsaJI were identified as potential restriction endonucleases to differentiate those meats. HindIII enzyme was unable to restrict the PCR product of both meats. The genetic differences within the cyt b gene among the two meats were successfully confirmed by PCR-RFLP analysis.
    Matched MeSH terms: DNA Restriction Enzymes
  4. Fulltext Overview of safety and efficacy of non-viral gene transfer in cartilage tissue engineering from the worldview of Islam
    Noorhidayah Md Nazir, Munirah Sha’ban
    International Medical Journal Malaysia, 2018;17(101):115-123.
    MyJurnal
    This paper examines the safety and efficacy of non-viral gene transfer in cartilage tissue engineering (TE) from the worldview of Islam. The first clinical trial treating adenosine deaminase deficient patients conducted in 1990 has triggered the development of gene transfer technology. The potential of gene transfer is further explored in TE field with the hope that it could prosper the regenerative medicine application. However, ethical issues become important when it comes to application of new treatment modalities, primarily in gene transfer because of genetic modification influences the basis of life - the DNA. Besides ethical issue, the application of gene transfer in treating diseases also attract views from religious context. The questions on the techniques to administer the gene in human, social acceptance of genetically modified cell and adverse effects from it are still debatable and unresolved. Apart from that dilemma, both safety and efficacy issues are raised due to the scientific uncertainty and social perception of the technology. Despite countless number of encouraging findings and recommendations by the proponents of
    the technology, gene transfer is currently available only in the research setting. The established guidelines are used to complement and provide the necessary foundations in discussing the aspects involved in the incorporation of gene transfer with cartilage TE. Relevant Islamic input are identified and aligned to those particular guidelines. It is hoped that the integration of Islamic inputs in the existing guidelines could suggest the safest approach in treating cartilage degenerative disease through gene transfer and TE.
    Matched MeSH terms: DNA
  5. Pengesahan bagi mengesan bakteria patogen makanan menggunakan teknologi mikroaturan DNA
    SAWEI J, NORRAKIAH ABDULLAH SANI, AMINAH ABDULLAH, SAHILAH ABD. MUTALIB
    Sains Malaysiana, 2013;42:1715-1720.
    Kajian ini dijalankan untuk mengesahkan kemampuan teknologi DNA mikroaturan cip gen OliproTM FoodPATH bagi mengesan bakteria patogen makanan. Sebanyak 9 jenis DNA bakteria patogen makanan telah digunakan iaitu Bacillus cereus, Escherichia coli O157:H7, Staphylococcus aureus, Vibrio cholerae, Vibrio parahaemolyticus, Listeria monocytogenes, Salmonella spp., Shigella spp. dan Campylobacter spp. Sebanyak 36 kombinasi templat DNA bakteria patogen makanan telah digunakan. Pengesahan bagi mengesan bakteria patogen makanan dilakukan dengan menggunakan kaedah reaksi berantai polimerase (PCR) dan penghibridan Southern-blotting di atas cip gen untuk mengesahkan kemampuannya. Keputusan daripada analisis hibridasi di atas cip gen telah dibandingkan dengan hasil gel elektroforesis 2.0% (w/v). Lima saringan diperlukan untuk menghabiskan 36 kombinasi templat DNA bakteria patogen makanan. Setiap saringan, satu cip gen telah digunakan sebagai kawalan negatif tidak diinokulasikan dengan sebarang kombinasi DNA bakteria patogen makanan. Daripada hasil kajian, didapati bahawa semua kombinasi templat DNA bakteria patogen makanan telah dapat dikesan. Cip yang digunakan sebagai kawalan negatif tidak menunjukkan kehadiran DNA. Oleh itu, daripada kajian ini cip gen OliproTM FoodPATH didapati memberikan keputusan yang lebih baik berbanding dengan 2.0% (w/v) gel elektroforesis.
    Matched MeSH terms: DNA
  6. Molecular Mitochondrial DNA and radiographic approaches for human archaeology identification
    Rus Dina Rus Din, Shahrul Hisham Zainal Ariffin, Sahidan Senafi, Rohaya Megat Abdul Wahab, Intan Zarina Zainol Abidin
    Sains Malaysiana, 2014;43:1523-1535.
    Ancient remains are considered very valuable artefacts, as they allow for the study of ancient cultures, phylogeny, evolution and the reconstruction of demographic history. To obtain all the information contained within remains, the investigation of such samples requires the expertise and various techniques from multiple fields of study. The present review focuses on the molecular biology and radiographic approaches used to identify ancient samples. Studies of ancient remains face various limitations; for example, the quality and quantity of the ancient samples can affect the difficulty of the investigations. Due to these limitations, new sophisticated techniques are being introduced to replace the earlier conventional techniques. A search was conducted using PubMed, Scopus, Science Direct and Science Finder to provide a new and timely review on the molecular mitochondrial DNA and radiographic analysis for human archaeology identification. The present review has determined that molecular biological approaches are very accurate and useful for the use in the ancestral determination of incomplete specimens, whereas observations of the dental pulp chamber are suitable for age at death estimations in both adults and children. However, these techniques are expensive and require expert personnel. Therefore, conventional approaches remain the favourite methods of most institutions, especially in Asia.
    Matched MeSH terms: DNA, Mitochondrial
  7. Phylogenetic relationships within the genus holothuria inferred from 16S mitochondiral rRNA gene sequences
    Kamarul Rahim Kamarudin, ‘Aisyah Mohamed Rehan, Ridzwan Hashim, Usup G, Maryam Mohamed Rehan
    Sains Malaysiana, 2016;45:1079-1087.
    This study aimed to resolve the taxonomic status of a morphologically undetermined sea cucumber species of order Apodida
    from Malaysia (GenBank accession no.: FJ223867) using partial 16S mitochondrial rRNA gene sequences and subsequently
    to determine the validity of morphological taxonomy of Holothuria species into its current subgenera. The undetermined
    species clustered with all taxa of Holothuria in previous study. Phylogenetic analyses using maximum parsimony and
    Bayesian methods suggest that the undetermined species was genetically closer to Holothuria (Lessonothuria) pardalis and
    Holothuria (Acanthotrapeza) coluber; and its position in both phylogenetic trees further suggests its status as a Holothuria
    taxon. Subgenera of Holothuria, Merthensiothuria and Metriatyla are monophyletic with strong bootstrap supports and
    posterior probabilities of clades, thus strengthening their morphological taxonomies. Nonetheless, the non-monophyly of
    subgenera of Halodeima, Microthele and Platyperona suggests a requirement for their taxonomic revisions using integrative
    taxonomy. The status of Holothuria (Halodeima) edulis subgroups in the maximum parsimony and Bayesian trees is
    indistinct and further taxonomic revisions are necessary. In terms of sister relationship, both phylogenetic trees suggest
    that subgenus Holothuria is a sister taxon of subgenus Roweothuria while the other sister relationships were unclear due
    to the undetermined species, paraphyly and polyphyly of a number of subgenera. Further studies with more specimens of
    genus Holothuria from broader geographical locations and various mtDNA genes along with morphological approaches
    may facilitate to provide better insights into the molecular phylogeny of subgenera of Holothuria.
    Matched MeSH terms: DNA, Mitochondrial
  8. Phylogeny of sea cucumber (Echinodermata: Holothuroidea) as inferred from 16S mitochondrial rRNa gene sequences
    Kamarul Rahim Kamarudin, Ridzwan Hashim, Usup G
    Sains Malaysiana, 2010;39:209-218.
    This study aimed to determine phylogenetic relationship between and among selected species of sea cucumbers (Echinodermata: Holothuroidea) using 16S mitochondrial ribosomal RNA (rRNA) gene. Phylogenetic analyses of 37 partial sequences of 16S mitochondrial rRNA gene using three main methods namely neighbour joining (NJ), maximum parsimony (MP) and maximum likelihood (ML) showed the presence of five main genera of sea cucumbers: Molpadia from order Molpadiida and four genera of order Aspidochirotida namely Holothuria, Stichopus, Bohadschia and Actinopyga. All of the 17 species obtained from Malaysia distributed among the main genera except within Actinopyga. Interestingly, Holothuria excellens was out of Holothuria group causing Holothuria to be paraphyletic. High bootstrap value and consistent clustering made Molpadia, Stichopus, Bohadschia and Actinopyga monophyletic. The relationship of Actinopyga with the other genera was unclarified and Stichopus was sister to Molpadia. The latter finding caused the resolution at order level unclear. The pairwise genetic distance calculated using Kimura 2-parameter model further supported and verified findings from the phylogenetic trees. Further studies with more samples and different mitochondrial DNA genes need to be done to get a better view and verification on the molecular phylogeny of sea cucumbers.
    Matched MeSH terms: DNA, Mitochondrial
  9. Fulltext Identification of Helicobacter pylori in Saliva of Patients with and without Gastritis by Polymerase Chain Reaction
    Sekhar Goud EVS, Kannan R, Rao UK, Joshua E, Tavaraja R, Jain Y
    J Pharm Bioallied Sci, 2019 Nov;11(Suppl 3):S523-S529.
    PMID: 31920269 DOI: 10.4103/jpbs.JPBS_260_18
    Aims and Objective: The aim of this study was to identify the presence of Helicobacter pylori in saliva of patients with and without gastritis by polymerase chain reaction (PCR) method.

    Materials and Methods: The study comprised 20 patients in Group I presenting with various symptoms of gastritis and 10 asymptomatic subjects in Group II. The intestinal endoscopy antral biopsies were collected from 20 symptomatic patients with gastroduodenal disorders. The saliva specimens were taken from all patients before endoscopy. PCR was performed using genomic DNA, isolated from the saliva and the biopsies of the patients as the template to detect the presence of the 16S ribosomal RNA gene in H. pylori.

    Results: In Group I, 10 (50%) cases of clinical gastritis were positive for H. pylori by endoscopy biopsy and 10 (50%) were negative. Of the 10 endoscopy biopsy positive cases for H. pylori, eight were PCR positive in saliva and two were negative. Of the 10 endoscopy biopsy negative cases, three were PCR positive for H. pylori in saliva and seven were negative. In Groups II, four were symptomatic for gastritis and six were negative. Of the six gastritis negative cases, three were PCR positive, four were gastritis positive, and three were PCR positive. Sensitivity and specificity of PCR were found to be 80% and 70%, respectively. The positive predictive and negative predictive values of PCR in saliva were 72.7% and 77.7%, respectively.

    Conclusion: PCR analysis of saliva may be handy in identification of H. pylori and serves as a noninvasive technique to diagnose and monitor the prognosis.

    Matched MeSH terms: DNA
  10. Fulltext Population structure and genetic diversity of Aedes aegypti and Aedes albopictus in Penang as revealed by mitochondrial DNA cytochrome oxidase I
    Md Naim D, Kamal NZM, Mahboob S
    Saudi J Biol Sci, 2020 Mar;27(3):953-967.
    PMID: 32127775 DOI: 10.1016/j.sjbs.2020.01.021
    The population genetics study is crucial as it helps in understanding the epidemiological aspects of dengue and help improving a vector control measures. This research aims to investigate the population genetics structure of two common species of Aedes mosquitoes in Penang; Aedes aegypti and Aedes albopictus using Cytochrome Oxidase I (COI) mitochondrial DNA (mtDNA) marker. Molecular investigations were derived from 440 bp and 418 bp mtDNA COI on 125 and 334 larvae of Aedes aegypti and Aedes albopictus respectively, from 32 locations in Penang. All samples were employed in the BLASTn for species identification. The haplotype diversity, nucleotide diversity, neutrality test and mismatch distribution analysis were conducted in DnaSP version 5.10.1. AMOVA analysis was conducted in ARLEQUIN version 3.5 and the phylogenetic reconstructions based on maximum likelihood (ML) and neighbor-joining (NJ) methods were implemented in MEGA X. The relationships among haplotypes were further tested by creating a minimum spanning tree using Network version 4.6.1. All samples were genetically identified and clustered into six distinct species. Among the species, Ae. albopictus was the most abundant (67.2%), followed by Ae. aegypti (25.2%) and the rest were counted for Culex sp. and Toxorhynchites sp. Both Ae. aegypti and Ae. albopictus show low nucleotide diversity (π) and high haplotype diversity (h), while the neutrality test shows a negative value in most of the population for both species. There are a total of 39 and 64 haplotypes recorded for Ae. aegypti and Ae. albopictus respectively. AMOVA analysis revealed that most of the variation occurred within population for both species. Mismatch distribution analysis showed bimodal characteristic of population differentiation for Ae. aegypti but Ae. albopictus showed unimodal characteristics of population differentiation. Genetic distance based on Tamura-Nei parameter showed low genetic divergent within population and high genetic divergent among population for both species. The maximum likelihood tree showed no obvious pattern of population genetic structure for both Ae. aegypti and Ae. albopictus from Penang and a moderate to high bootstrap values has supported this conclusion. The minimum spanning network for Ae. aegypti and Ae. albopictus showed five and three dominant haplotypes respectively, which indicates a mixture of haplotypes from the regions analysed. This study revealed that there is no population genetic structure exhibited by both Ae. aegypti and Ae. albopictus in Penang. Mutation has occurred rapidly in both species and this will be challenging in controlling the populations. However, further analysis needed to confirm this statement.
    Matched MeSH terms: DNA, Mitochondrial
  11. First Report of Rubber Tree Stem Rot Caused by Fusarium oxysporum in China
    Li BX, Shi T, Liu XB, Lin CH, Huang GX
    Plant Dis, 2014 Jul;98(7):1008.
    PMID: 30708897 DOI: 10.1094/PDIS-01-14-0004-PDN
    Rubber tree (Hevea brasiliensis) is an important crop in tropical regions of China. In October 2013, a new stem rot disease was found on cv. Yunyan77-4 at a rubber tree plantation in Hekou, Yunnan Province. There were about 100 plants, and diseased rubber trees accounted for 30% or less. Initially, brown-punctuate secretion appeared on the stem, which was 5 to 6 cm above the ground. Eventually, the secretion became black and no latex produced from the rubber tree bark. After removing the secretion, the diseased bark was brown putrescence, but the circumambient bark was normal. Upon peeling the surface bark, the inner bark and xylem had brown rot and was musty. The junction between health and disease was undulate. On the two most serious plants, parts of leaves on the crown were yellow, and the root near the diseased stem was dry and puce. The pathogen was isolated and designated HbFO01; the pathogenicity was established by following Koch's postulates. The pathogen was cultivated on a potato dextrose agar (PDA) plate at 28°C for 4 days. Ten plants of rubber tree cv. Yunyan77-4 were selected from a disease-free plantation in Haikou, Hainan Province, and the stem diameter was about 7 cm. The bark of five plants was peeled, and one mycelium disk with a diameter of 1 cm was inserted into the cut and covered again with the bark. The other five plants were treated with agar disks as controls. The inoculation site was kept moist for 2 days, and then the mycelium and agar disk were removed. On eighth day, symptoms similar to the original stem lesions were observed on stems of inoculated plants, while only scars formed on stems of control plants. The pathogen was re-isolated from the lesions of inoculated plants. On PDA plates, the pathogen colony was circular and white with tidy edges and rich aerial hyphae. Microscopic examination showed microconidia and chlamydospores were produced abundantly on PDA medium. The falciform macroconidia were only produced on lesions and were slightly curved, with a curved apical cell and foot shaped to pointed basal cell, usually 3-septate, 16.2 to 24.2 × 3.2 to 4.0 μm. Microconidia were produced in false heads, oval, 0-septate, 6.2 to 8.2 × 3.3 to 3.8 μm, and the phialide was cylindrical. Chlamydospores were oval, 6.4 to 7.2 × 3.1 to 3.8 μm, alone produced in hypha. Morphological characteristics of the specimen were similar to the descriptions for Fusarium oxysporum (2). Genomic DNA of this isolate was extracted with a CTAB protocol (4) from mycelium and used as a template for amplification of the internal transcribed spacer (ITS) region of rDNA with primer pair ITS1/ITS4 (1). The full length of this sequence is 503 nt (GenBank Accession No. KJ009335), which exactly matched several sequences (e.g., JF807394.1, JX897002.1, and HQ451888.1) of F. oxysporum. Williams and Liu had listed F. oxysporum as the economically important pathogen of Hevea in Asia (3), while this is, to our knowledge, the first report of stem rot caused by F. oxysporum on rubber tree in China. References: (1) D. E. L. Cooke et al. Fungal Genet. Biol. 30:17, 2000. (2) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual, 2006. (3) T. H. Williams and P. S. W. Liu. A host list of plant diseases in Sabah, Malaysia, 1976. (4) J. R. Xu et al. Genetics 143:175, 1996.
    Matched MeSH terms: DNA, Ribosomal
  12. Occurrence of Anthracnose Caused by Colletotrichum truncatum on Chickpea (Cicer arietinum) in Malaysia
    Mahmodi F, Kadir JB, Nasehi A, Puteh A, Soleimani N
    Plant Dis, 2013 Nov;97(11):1507.
    PMID: 30708462 DOI: 10.1094/PDIS-03-13-0231-PDN
    At least nine Colletotrichum species, particularly Colletotrichum truncatum, have been recorded on legumes worldwide (1). In June 2010, samples of chickpea leaflets showing leaf spot disease symptoms were collected from experimental farms in Ladang Dua, Selangor state of Malaysia. Tan lesions with darker brown borders were observed on leaflets and were associated with premature leaf drop. Stem lesions initially appeared on the lower parts of stems and later progressed higher in the plant. Lesions often girdled the stem and caused severe dieback. Abundant acervuli developed in the lesions visible as black dots. Foliar lesions were removed, surface sterilized in 1% sodium hypochlorite for 2 min, rinsed twice with distilled water, dried on sterilized tissue paper, plated on PDA plates, and incubated at 25°C (3). Three isolates of the fungus were obtained and identified as C. truncatum on the basis of morphological characteristics (2). The isolates were deposited in the University Putra of Malaysia Culture Collection (UPMCC). Colony characteristics on PDA varied from greyish white to dark in color and exhibited mycelial growth with sparse acervuli. The isolates produced both sclerotia and setae in culture. Conidia (mean ± SD = 22 ± 0.83 × 3.6 ± 0.08 μm, L/W ratio = 6.1) produced in acervuli were falcate, hyaline, and aseptate, with tapering towards the acute and greatly curved apex. The conidial mass color varied from pale buff to saffron. Isolates produced simple to slightly lobed, mainly short clavate appressoria (mean ± SD = 9.60 ± 0.36 × 6.67 ± 0.29 μm, L/W ratio = 1.45). Amplification and sequence analysis of coding and none-coding regions of the ITS-rDNA (GenBank Accession JX971160), actin (JX975392), β-tubulin (KC109495), histone (KC109535), chitin synthase (KC109575), and glyceraldehyde-3-phosphate dehydrogenase (KC109615) obtained from the representative isolate, CTM37, aligned with deposited sequences from GenBank and revealed 99 to 100% sequence identity with C. truncatum strains (AJ301945, KC110827, GQ849442, GU228081, GU228359, and HM131501 from GenBank). Isolate CTM37 was used to test pathogenicity in the greenhouse. Five chickpea seeds of cultivar ILC-1929 were sown per pot in four replications. Ten days after seedling emergence, plants were inoculated with a spore suspension (concentration = 106 conidia ml-1) and check pots were sprayed with distilled water. After inoculation, the plants were covered with plastic bags for 48 h and kept at 28 to 33°C and >90% RH. After incubation, the plastic bags were removed and the plants were placed on greenhouse benches and monitored daily for symptom development (3). One week after inoculation, typical anthracnose symptoms developed on the leaves and stems of inoculated plants including acervuli formation, but not on the checks. A fungus with the same colony and conidial morphology as CTM37 was recovered from the lesions on the inoculated plants. The experiment was repeated twice. The ability to accurately diagnose Colletotrichum species is vital for the implementation of effective disease control and quarantine measures. We believe this is the first report of C. truncatum causing anthracnose on chickpea in Malaysia. References: (1) B. D. Gossen et al. Can. J. Plant Pathol. 31:65, 2009. (2) B. C. Sutton. The Genus Glomerella and its anamorph Colletotrichum. CAB International, Wallingford. UK. 1992. (3) P. P. Than et al. Plant Pathol. 57:562, 2008. ERRATUM: A correction was made to this Disease Note on May 19, 2014. The author N. Soleimani was added.
    Matched MeSH terms: DNA, Ribosomal
  13. First Report of Bark Dieback on Blueberry Caused by Botryosphaeria dothidea in Korea
    Choi IY
    Plant Dis, 2011 Feb;95(2):227.
    PMID: 30743439 DOI: 10.1094/PDIS-05-10-0371
    This study was conducted to identify the causal organism of bark dieback disease of highbush blueberry (Vaccinium corymbosum L.) observed in Korea. Blueberry, a woody plant that is native to North America, belongs to the family Ericaceae and genus Vaccinium. Of the 400 species of blueberry in the world, most are distributed in the tropics of Malaysia and Southeast Asia. Highbush blueberry is abundantly grown in Canada and the United States and has become a popular commercial crop in Korea for products such as jam, wine, and sauce. Bark dieback disease of blueberry was found in Sunchang (<5% incidence), Jeollabuk-do, Korea in July 2009. Typical symptoms of the disease were blight and dieback on the stems with lesions extending along entire branches. Morphological examination revealed that the perithecia were of the globose type with a nipple, 155 to 490 (374.6) μm, and brown on the dead bark. Asci were bitunicate and clavate or cylindrical with dimensions of 63 to 125 × 16 to 20 μm and containing eight ascospores. Ascospores were of the long ovoid type with dimensions of 13.2 to 23.7 (17.98) × 25.4 to 41.1 (33.21) μm. From extracted genomic DNA, the internal transcribed spacer (ITS)-5.8S ribosomal DNA region was amplified with universal primers ITS1 (5'-TCCGTAGGTGAACCTGCGG-3') and ITS4 (5'-TCCTCCGCTTATTGATATGC-3'). A BLAST search of GenBank with the ITS sequence revealed that the Sunchang isolate (GenBank Accession No. HQ384217) had 99 to 100% sequence identity with the following Botryosphaeria dothidea accessions: FJ517657, AJ938005, FJ478129, FJ171723, and AJ938004. Phylogenetic analysis with the Sunchang isolate, B. dothidea strains, and related species revealed that the B. dothidea isolate and strains comprised a monophyletic group distinguished from other Botryosphaeria spp. including B. ribis, B. parva, B. protearum, B. lutea, B. australis, B. rhodina, B. obtuse, and B. stevensii (2). On the basis of morphological and molecular results, the isolate was identified as B. dothidea (Moug.) Ces. & De Not. A culture of B. dothidea isolate was grown on potato dextrose agar (PDA) for 10 days. A 5-mm plug was inoculated into stem wounds created with a No. 2 cork borer in 20 2-year-old disease-free blueberry plants grown in a greenhouse. Six plants inoculated with only PDA plugs served as noninoculated controls. The wounds were covered with Parafilm. After 3 months, the Parafilm was removed and black lesions were observed at the fungal inoculation sites, while no lesion was observed on the control plants. To complete Koch's postulates, the fungus was reisolated from the lesions and confirmed to be B. Dothidea (1). There is an urgent need to determine the spread of this disease in Korea, estimate the losses, and develop methods for reducing damage through biological and eco-friendly cultural control methods. References: (1) D. Jurc et al. Plant Pathol. 55:299, 2006. (2) B. Slippers et al. Mycologia 96:83, 2004.
    Matched MeSH terms: DNA, Ribosomal Spacer
  14. First Report of Podosphaera fusca Causing Powdery Mildew of Cosmos caudatus in Malaysia
    Siddiqui Y, Sariah M, Kausar H
    Plant Dis, 2011 Apr;95(4):495.
    PMID: 30743349 DOI: 10.1094/PDIS-12-10-0866
    Cosmos caudatus Kunth. (Asteraceae), commonly known as ulam raja, is widely grown as an herbal aromatic shrub. In Malaysia, its young leaves are popularly eaten raw as salad with other greens and have been reported to possess extremely high antioxidant properties, which may be partly responsible for some of its believed medicinal functions. In early 2010, a suspected powdery mildew was observed on ulam raja plants at the Agricultural Park of Universiti Putra Malaysia. Initially, individual, white, superficial colonies were small and almost circular. Later, they enlarged and coalesced to cover the whole abaxial leaf surface. With development of the disease, all green parts (leaves, stems, and petioles) became covered with a continuous mat of mildew, giving a dusty appearance. Newly emerged leaves rapidly became infected. Diseased leaves ultimately senesced and dried up, making them aesthetically unattractive and unmarketable. The pathogen produced conidia in short chains (four to six conidia) on erect conidiophores. Conidiophores were unbranched, cylindrical, 125 to 240 μm long, with a slightly swollen foot cell. Individual conidia were hyaline, ellipsoid, and 25 to 30 (27.5) × 15 to 20 (17.5) μm with fibrosin inclusions. Morphological descriptions were consistent with those described for Sphaerotheca fuliginea or S. fusca, which has lately been reclassified as Podosphaera fusca (1). From extracted genomic DNA of P. fusca UPM UR1, the internal transcribed spacer (ITS) region was amplified with ITS1 (5'-TCCGTAGGTGAACCTGCGG-3') and ITS4 (5'-TCCTCCGCTTATTGATATGC-3'). A BLAST search of GenBank with an ITS rDNA sequence of this fungus (GenBank Accession No. HQ589357) showed a maximum identity of 98% to the sequences of two P. fusca isolates (GenBank Accession Nos. AB525915.1 and AB525914.1). To satisfy Koch's postulates, the pathogenicity of fungal strain UPM UR1 was verified on 4-week-old plants. Inoculation was carried out by gently rubbing infected leaves onto healthy plants of C. caudatus. Ten pots of inoculated plants were kept under a plastic humid chamber and 10 pots of noninoculated plants, placed under another chamber, served as controls. After 48 h, the plants were then placed under natural conditions (25 to 28°C). Powdery mildew symptoms, similar to those on diseased field plants, appeared after 7 days on all inoculated plants. The white, superficial colonies enlarged and merged to cover large areas within 2 weeks. The infected leaf tissues became necrotic 6 to 8 days after the appearance of the first symptoms. Sporulation of P. fusca was observed on all infected leaves and stems. No symptoms were seen on the control plants. To our knowledge, this is the first report of P. fusca causing powdery mildew on C. caudatus in Malaysia. This pathogen has also been reported previously to be economically important on a number of other hosts. With ulam raja plants, more attention should be given to prevention and control measures to help manage this disease. Reference: (1) U. Braun and S. Takamatsu. Schlechtendalia 4:1, 2000.
    Matched MeSH terms: DNA, Ribosomal
  15. First Report of Alternaria tenuissima Causing Leaf Spot on Eggplant in Malaysia
    Nasehi A, Kadir JB, Abidin MAZ, Wong MY, Mahmodi F
    Plant Dis, 2012 Aug;96(8):1226.
    PMID: 30727083 DOI: 10.1094/PDIS-03-12-0237-PDN
    A leaf spot on eggplant (Solanum melongena) was observed in major eggplant growing regions in Malaysia, including the Cameron Highlands and Johor State, during 2011. Disease incidence averaged approximately 30% in severely infected regions in about 150 ha of eggplant fields and greenhouses examined. Early symptoms consisted of small, circular, brown, necrotic spots uniformly distributed on leaves. The spots gradually enlarged and developed concentric rings. Eventually, the spots coalesced and caused extensive leaf senescence. A fungus was recovered consistently by plating surface-sterilized (1% NaOCl) sections of symptomatic leaf tissue onto potato dextrose agar (PDA). For conidial production, the fungus was grown on potato carrot agar (PCA) and V8 agar media under a 16-h/8-h dark/light photoperiod at 25°C (4). Fungal colonies were a dark olive color with loose, cottony mycelium. Simple conidiophores were ≤120 μm long and produced numerous conidia in long chains. Conidia averaged 20.0 × 7.5 μm and contained two to five transverse septa and the occasional longitudinal septum. Twelve isolates of the fungus were identified as Alternaria tenuissima on the basis of morphological characterization (4). Confirmation of the species identification was obtained by molecular characterization of the internal transcribed spacer (ITS) region of rDNA amplified from DNA extracted from a representative isolate using universal primers ITS4 and ITS5 (2). The 558 bp DNA band amplified was sent for direct sequencing. The sequence (GenBank Accession No. JQ736021) was subjected to BLAST analysis (1) and was 99% identical to published ITS rDNA sequences of isolates of A. tenuissima (GenBank Accession Nos. DQ323692 and AY154712). Pathogenicity tests were performed by inoculating four detached leaves from 45-day-old plants of the eggplant cv. 125066x with 20 μl drops (three drops/leaf) of a conidial suspension containing 105 conidia/ml in sterile distilled water. Four control leaves were inoculated with sterile water. Leaves inoculated with the fungus and those treated with sterile water were incubated in chambers at 25°C and 95% RH with a 12-h photoperiod/day (2). Leaf spot symptoms typical of those caused by A. tenuissima developed on leaves inoculated with the fungus 7 days after inoculation, and the fungus was consistently reisolated from these leaves. The control leaves remained asymptomatic and the pathogen was not reisolated from the leaves. The pathogenicity test was repeated with similar results. To our knowledge, this is the first report of A. tenuissima causing a leaf spot on eggplant in Malaysia. A. tenuissima has been reported to cause leaf spot and fruit rot on eggplant in India (3). References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) B. M. Pryor and T. J. Michailides. Phytopathology 92:406, 2002. (3) P. Raja et al. New Disease Rep. 12:31, 2005. (4) E. G. Simmons. Page 1 in: Alternaria Biology, Plant Diseases and Metabolites. J. Chelchowski and A. Visconti, eds. Elsevier, Amsterdam, 1992.
    Matched MeSH terms: DNA, Ribosomal
  16. First Report of a Soft Rot of Phalaenopsis aphrodita Caused by Dickeya dieffenbachiae in China
    Zhou JN, Lin BR, Shen HF, Pu XM, Chen ZN, Feng JJ
    Plant Dis, 2012 May;96(5):760.
    PMID: 30727539 DOI: 10.1094/PDIS-11-11-0942
    Phalaenopsis orchids, originally from tropical Asia, are mainly planted in Thailand, Singapore, Malaysia, the Philippines, and Taiwan and have gained popularity from consumers all over the world. The cultivation area of Phalaenopsis orchids has been rising and large-scale bases have been established in mainland China, especially South China because of suitable environmental conditions. In September 2011, a soft rot of Phalaenopsis aphrodita was found in a Phalaenopsis planting base in Guangzhou with an incidence of ~15%. Infected plants initially showed water-soaked, pale-to-dark brown pinpoint spots on leaves that were sometimes surrounded by a yellow halo. Spots expanded rapidly with rising humidity and temperatures, and in a few days, severely extended over the blade with a light tan color and darker brown border. Lesions decayed with odorous fumes and tissues collapsed with inclusions exuding. The bacterium advanced to the stem and pedicle. Finally, leaves became papery dry and the pedicles lodged. Six diseased samples were collected, and bacteria were isolated from the edge of symptomatic tissues after sterilization in 0.3% NaOCl for 10 min, rinsing in sterile water three times, and placing on nutrient agar for culture. Twelve representative isolates were selected for further characterization. All strains were gram negative, grew at 37°C, were positive for indole production, and utilized malonate, glucose, and sucrose but not glucopyranoside, trehalose, or palatinose. Biolog identification (version 4.20.05, Hayward, CA) was performed and Pectobacterium chrysanthemi (SIM 0.868) was confirmed for the tested isolates (transfer to genus Dickeya). PCR was used to amplify the 16S rDNAgene with primers 27f and 1492r, dnaX gene with primers dnaXf and dnaXr (3), and gyrB gene with primers gyrBf (5'-GAAGGYAAAVTKCATCGTCAGG-3') and gyrB-r1 (5'-TCARATATCRATATTCGCYGCTTTC-3') designed on the basis of the published gyrB gene sequences of genus Dickeya. BLASTn was performed online, and phylogeny trees (100% bootstrap values) were created by means of MEGA 5.05 for these gene sequences, respectively. Results commonly showed that the representative tested strain, PA1, was most homologous to Dickeya dieffenbachiae with 98% identity for 16S rDNA(JN940859), 97% for dnaX (JN989971), and 96% for gyrB (JN971031). Thus, we recommend calling this isolate D. dieffenbachiae PA1. Pathogenicity tests were conducted by injecting 10 P. aphrodita seedlings with 100 μl of the bacterial suspension (1 × 108 CFU/ml) and another 10 were injected with 100 μl of sterile water as controls. Plants were inoculated in a greenhouse at 28 to 32°C and 90% relative humidity. Soft rot symptoms were observed after 2 days on the inoculated plants, but not on the control ones. The bacterium was isolated from the lesions and demonstrated identity to the inoculated plant by the 16S rDNA sequence comparison. Previously, similar diseases of P. amabilis were reported in Tangshan, Jiangsu, Zhejiang, and Wuhan and causal agents were identified as Erwinia spp. (2), Pseudomonas grimontii (1), E. chrysanthemi, and E. carotovora subsp. carovora (4). To our knowledge, this is the first report of D. dieffenbachiae causing soft rot disease on P. aphrodita in China. References: (1) X. L. Chu and B. Yang. Acta Phytopathol. Sin. 40:90, 2010. (2) Y. M. Li et al. J. Beijing Agric. Coll. 19:41, 2004. (3) M. Sławiak et al. Eur. J. Plant Pathol. 125:245, 2009. (4) Z. Y. Wu et al. J. Zhejiang For. Coll. 27:635, 2010.
    Matched MeSH terms: DNA, Ribosomal
  17. Differentiation of Ficus deltoidea varieties and chemical marker determination by UHPLC-TOFMS metabolomics for establishing quality control criteria of this popular Malaysian medicinal herb
    Afzan A, Kasim N, Ismail NH, Azmi N, Ali AM, Mat N, et al.
    Metabolomics, 2019 Mar 04;15(3):35.
    PMID: 30830457 DOI: 10.1007/s11306-019-1489-2
    BACKGROUND: Ficus deltoidea Jack (Moraceae) is a plant used in Malaysia for various diseases including as a supplement in diabetes management. Morphology distinction of the 7 main varieties (var. angustifolia, var. bilobata, var. deltoidea, var. intermedia, var. kunstleri, var. motleyana and var. trengganuensis) is challenging due to the extreme leaf heterophylly and unclear varietal boundaries, making it difficult for quality control of F. deltoidea products.

    OBJECTIVE: We aimed to compare the phytochemical composition of 7 varieties growing in different conditions at various geographical locations. We also aimed to establish the quality control markers for the authentication of these varieties.

    METHODS: We applied untargeted UHPLC-TOFMS metabolomics to discriminate 100 leaf samples of F. deltoidea collected from 6 locations in Malaysia. A genetic analysis on 21 leaf samples was also performed to validate the chemotaxonomy differentiation.

    RESULTS: The PCA and HCA analysis revealed the existence of 3 chemotypes based on the differentiation in the flavonoid content. The PLS-DA analysis identified 15 glycosylated flavone markers together with 1 furanocoumarin. These markers were always consistent for the respective varieties, regardless of the geographical locations and growing conditions. The chemotaxonomy differentiation was in agreement with the DNA sequencing. In particular, var. bilobata accession which showed divergent morphology was also differentiated by the chemical fingerprints and genotype.

    CONCLUSION: Chemotype differentiation based on the flavonoid fingerprints along with the proposed markers provide a powerful identification tool to complement morphology and genetic analyses for the quality control of raw materials and products from F. deltoidea.

    Matched MeSH terms: Sequence Analysis, DNA
  18. Fulltext Simvastatin Inhibits Endotoxin-Induced Apoptosis in Liver and Spleen Through Up-Regulation of Survivin/NF-κB/p65 Expression
    Nežić L, Amidžić L, Škrbić R, Gajanin R, Nepovimova E, Vališ M, et al.
    Front Pharmacol, 2019;10:54.
    PMID: 30828299 DOI: 10.3389/fphar.2019.00054
    Endotoxemia is associated by dysregulated apoptosis of immune and non-immune cells. We investigated whether simvastatin has anti-apoptotic effects, and induces hepatocytes and lymphocytes survival signaling in endotoxin-induced liver and spleen injuries. Wistar rats were divided into the groups pretreated with simvastatin (20 or 40 mg/kg, orally) prior to a non-lethal dose of lipopolysaccharide (LPS), the LPS group, and the control. The severity of tissue inflammatory injuries was expressed as hepatic damage scores (HDS) and spleen damage scores (SDS), respectively. The apoptotic cell was detected by TUNEL (Terminal deoxynucleotidyl transferase dUTP Nick End Labeling) and immunohistochemical staining (expression of cleaved caspase-3, and anti-apoptotic Bcl-xL, survivin and NF-κB/p65). Simvastatin dose-dependently abolished HDS and SDS induced by LPS (p < 0.01), respectively. Simvastatin 40 mg/kg significantly decreased apoptotic index and caspase-3 cleavage in hepatocytes and lymphocytes (p < 0.01 vs. LPS group, respectively), while Bcl-XL markedly increased accordingly with simvastatin doses. In the simvastatin, groups were determined markedly increased cytoplasmic expression of survivin associated with nuclear positivity of NF-κB, in both hepatocytes and lymphocytes (p < 0.01 vs. LPS group). Cell-protective effects of simvastatin against LPS seemed to be mediated by up-regulation of survivin, which leads to reduced caspase-3 activation and inhibition of hepatocytes and lymphocytes apoptosis.
    Matched MeSH terms: DNA Nucleotidylexotransferase
  19. Description of the female of Simulium (Gomphostilbia) aziruni (Diptera: Simuliidae) and its genetic relationships with members of the Simulium gombakense species-group
    Ya'cob Z, Takaoka H, Low VL, Tan TK, Sofian-Azirun M
    Acta Trop, 2019 May;193:66-70.
    PMID: 30807749 DOI: 10.1016/j.actatropica.2019.02.023
    Simulium (Gomphostilbia) aziruni Takaoka, Hashim & Chen was described initially based only on a pupa and a mature larva collected from Peninsular Malaysia. Herein, we describe the morphological characters of the female of S. aziruni for the first time. It resembles those of the other members of the Simulium gombakense species-group by the genital fork with a distinct projection directed medioposteriorly from each arm and claw with a large basal tooth. Cytochrome c oxidase I (COI) barcoding analysis indicates that S. aziruni is the sister species of S. maleewongae, but both are distantly separated by a genetic distance of 4.9%.
    Matched MeSH terms: DNA Barcoding, Taxonomic
  20. Fulltext Prevalence of obesity in Malaysia: perspectives in Terengganu
    Muhammad, I. N., Saifullah, K., Hassan, B., Yasrul, I., Norizan, A./G., Ahmed Zubaidi, A. L., et al.
    Asian Journal of Medicine and Biomedicine, 2018;2(1):0-0.
    MyJurnal
    A normal health status is highly depends on body weight. Many health problems may occur or impose risk for
    extremely obese individuals. Etiopathology of obesity includes interaction of several factors inclusive of
    genetic and non-genetic factors such as lifestyle changes. This study aimed to launch Malaysian Obesity
    DNA Bank and determine the prevalence of obesity along with anthropometric measurements of the subjects.
    The cross-sectional study was conducted on total of 340 subjects (obese = 95, overweight = 122 and normal =
    123), aged 19-60 years, in Terengganu. The BMI and well appropriate anthropometric measurements (waist
    circumference, hip circumference, waist to hip ratio, fat percentage and ASindex) were determined through
    standard protocols and formulae. The mean difference of anthropometrics was determined by independent ttest. Data was analysed using SPSS ver.16.0.0. The BMI was determined for all subjects and it was found that
    out of 340 subjects, a total of 95 (27.9%) subjects were obese , followed by a total of 122 (35.8%) subjects
    were overweight and normal individuals were 123 (36.1%). The mean of the BMI, WHR, Fat% and ASindex,
    in Malay obese were 32.83, 0.88, 33.5 and 13.21 respectively, while in normal healthy individuals were 22.1,
    0.78, 24.2 and 20.1 respectively. The difference of mean of BMI, WHR, Fat% and ASindex was calculated to
    be 10.73, 0.1, 9.3 and 6.89 respectively. To the best of our knowledge, this is the first report in Malaysia,
    reporting that this is very first Obesity DNA Bank in South East Asia region and prevalence of obesity in
    Terengganu, Malaysia to be 27.9%. In addition, it indicates a significant mean difference for anthropometric
    measurements among obese and normal individuals. For Asindex calculations suggest that the prevalence of
    genocide obesity is greater 89.9% of android obesity in Malay obese attributes
    .
    Matched MeSH terms: DNA
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