Kajian ini dijalankan bagi menentukan kesan penambahan kalsium karbonat (CaCO3) dan sukrosa ke atas ekstrak roselle dengan menggunakan Kaedah Respon Permukaan. Sebanyak 21 perlakuan digunakan berdasarkan reka bentuk eksperimen Central Composite Rotatable Design (CCRD). Penentuan nilai pH dan Briks telah dijalankan. Bagi nilai pH, model yang didapati adalah 4.03+0.39x1-8.20x2-0.05x1 2-4.16x2 2-0.02x1x2 dengan nilai x1 adalah kalsium karbonat dan nilai x2 adalah sukrosa. Nilai positif pada kepekatan kalsium menunjukkan penambahan kalsium meningkatkan nilai pH ekstrak roselle. Model untuk Briks adalah 8.91 + 0.23x1 + 2.20x2. Nilai positif pada kepekatan kalsium dan sukrosa menunjukkan penambahan kalsium dan sukrosa meningkatkan nilai Briks ekstrak roselle. Penilaian sensori dengan menggunakan panel terlatih (n=8) menunjukkan penambahan sukrosa meningkatkan penerimaan keseluruhan terhadap ekstrak Roselle. Manakala, kesan penambahan kalsium karbonat telah mengurangkan penerimaan keseluruhan kerana ia merendahkan rasa masam.
Kekasaran permukaan amat mempengaruh tingkah-laku ricihan dan nilai sudut geseran puncak sesuatu satah ketakselanjaran. Oleh yang demikian, kekasaran permukaan merupakan faktor yang penting yang perlu dinilai dalam menentukan kestabilan struktur pengorekan dalam jasad batuan. Beberapa kaedah boleh digunakan untuk menentukan kekasaran permukaan ketakselanjaran. Kertas ini memperkenalkan satu pendekatan yang mudah dalam bentuk dua rumus polinominal yang boleh digunakan untuk mengkorelasi sudut geseran puncak ketakselanjaran (φp) batuan syis segar dan terluluhawa sedikit dengan pekali kekasaran kekar, PKK. Rumus polinominal ini ialah φp= -0.022PKK2 + 3.21PKK + 28.1 untuk ketakselanjaran segar dan φp= -0.025PKK2 + 3.24PKK + 26.6 untuk ketakselanjaran terluluhawa sedikit. Kedua-dua rumus ini memperlihatkan nilai pekali penentuan, R2 = 0.98. Hasil kajian ini menawarkan satu kaedah alternatif penganggaran φp satah ketakselanjaran daripada nilai PKK yang diukur pada permukaan ketakselanjaran di lapangan.
Kajian mengenai taburan dan kelimpahan parasitoid lalat rumah (Musa domestica) telah dijalankan di ladang ternakan ayam di beberapa negeri di Semenanjung Malaysia. Didapati bahawa spesies parasitoid yang memparasit pupa lalat di ladang ternakan ayam di Semenanjung Malaysia adalah Splangia endius, Splangia cameroni, Dirhinus himalayanus
dan Pachycrepoideus vindemiae. Taburan populasi S. endius dan D. himalayanus adalah menyeluruh iaitu terdapat di semua negeri berbanding P. vindemiae. Namun, kelimpahan (bilangan individu) S. endius adalah tinggi secara signifikan (p<0.05) berbanding lain-lain spesies parasitoid. Kelimpahan D. himalayanus, S. cameroni dan P. vindemiae adalah tidak berbeza secara signifikan antara negeri. Ini menunjukkan S. endius adalah berpotensi untuk dijadikan agen kawalan biologi lalat rumah di ladang ternakan ayam di Malaysia. Kelimpahan parasitoid adalah tinggi secara signifikan (p<0.05) di ladang ayam telur berbanding ladang ayam daging. Secara keseluruhannya, terdapat perbezaan yang tinggi dan signifikan tentang populasi parasitoid di Johor berbanding Kelantan tetapi tidak (p<0.05) di antara negeri Johor, Selangor, Pulau Pinang dan Pahang. Perbezaan yang tidak signifikan juga direkodkan antara kelimpahan pupa lalat rumah dan parasitoid di sepanjang tempoh kajian dan antara lokasi di Malaysia. Kesan suhu dan jumlah hujan
didapati tidak mempengaruhi perbezaan kelimpahan parasitoid di ladang ayam. Potensi S. endius dan D. himalayanus untuk dikomersialkan pengeluarannya juga dibincangkan.
The aim of this study was to determine the inhibition effects of the terpenoid of Plantago major on calcium oxalate crystals in vitro and to compare the effects of Plantago major with clinically used drugs like zyloric and potassium citrate for the treatment of urinary stone. Modified Schneider slide gel method was used for in vitro study and crystals formed were measured by Image Analyser System (Leica) after 24 h of treatment. The active compound in the methanol extract of Plantago major was isolated by bioassay - guided fractionation & isolation method. Dimethylsulphoxide (DMSO) was used as the negative control and zyloric and potassium citrate were used as positive controls. The results showed that crude methanol extract of Plantago major contained the active compound terpenoid. Terpenoid, zyloric and potassium citrate at concentrations in the range of (100 μg/mL - 250 μg/mL) significantly inhibited the area of crystal formation in comparison to the negative control after 24 h (p<0.001). The Zyloric and terpenoid of Plantago major in the concentrations of (100 μg/mL-250 μg/mL) inhibited the sizes of crystals significantly (p<0.05). Potassium citrate was more effective, than terpenoid of Plantago major in inhibiting the size of crystals at two concentrations i.e. 100 μg/mL and 150 μg/mL respectively (p<0.05). However the IC50 values for terpenoid of Plantago major, potassium citrate and zyloric were 250 μg/mL, 300 μg/mL and 550 μg/mL, respectively. The inhibition effect of the terpenoid of Plantago major extract on crystal size was much better than Zyloric and potassium citrate.
Science curriculum that emphasizes science process skills (SPS) will be able to help students acquire and understand information, as well as improve skills in critical thinking and decision making. Consequently, the SPS should be systematically taught to the students from as early as at primary school. To produce students who acquire the science process skills, the teacher should be competent in SPS; theoretically and practically. Thus, the aim of this study was to identify the teachers’ perception on their understanding of integrated SPS, conceptual knowledge and the competence level of integrated SPS of primary school teachers in Kuala Lumpur. A questionnaire was administered to 329 science teachers from 52 primary schools that have been selected randomly. The reliability of the instrument is 0.80. The data were analyzed using descriptive and inferential analysis (t-test and ANOVA). The findings showed that majority of teachers (74.1% to 83.6%) perceived that they have a high level of understanding on each of the sub-component of an integrated SPS. However teachers’ perception on their level of understanding of the integrated SPS was found to be inconsistent with the actual level of understanding (conceptual knowledge). Teachers had a low level of conceptual understanding
of the integrated SPS. The findings also revealed that science teachers did not have sufficient conceptual knowledge of integrated SPS to teach their students to understand it in a meaningful way. However, the competency level of the integrated SPS among teachers is high with a mean of 20.86. As a conclusion, the study showed that teachers competency in the integrated SPS is good at the practical stage but not theoretically. Therefore, emphasis should be given to integrated SPS both conceptual and operational knowledge in pre and in-service training to ensure that the teachers understand, acquire and are able to implement the skills meaningfully.
There has been extensive research to look at the pharmacological effects on immediate recovery after stroke. However, there is lack of research to investigate the contributions of other effects that influence recovery after stoke. The study aims is to determine the extent of psychological factors such as illness representations, perceived control, psychological distress and posttraumatic stress symptoms in predicting recovery from disability following stroke. A cross sectional and longitudinal study with two times lines (six weeks and six months post−stroke) will be conducted in this study. It will consists of 180 patients from UKM Medical Centre. The idea of enhancing recovery from disability through the usage of psychological variables is important to assist recovery despite depending on a fully medical approach. Hence, it will provide information to the healthcare services and the government to improve the services and treatment intervention of the stroke sufferers.
This study aimed to investigate the usage of Behavioural Cognitive approach in group counselling on social anxiety among adolescents. This study also identified social anxiety sympthoms and social situations that cause anxiety among them. A total of 72 secondary school students from one particular school in Terengganu participated in this experimental study. Data was analysed using descriptive statistics and ANOVA test. Results revealed that there was a significant difference in the mean score of Social Anxiety Scale for Adolesence – SASA in the post test between the control and treatment group, F(1,70) =1.068, p<0.05. This indicates that the Cognitive Behavioural approach in group counselling was found to be effective in reducing social anxiety among adolescents in group counselling. It was also found that the sympthoms that caused social anxiety included having rapid heart beat, difficulty in giving attention, experincing chills, and empty thoughts. The social situations that were found to be significant in increasing social anxiety were when they felt being observed by others, doing something in public, talking or giving a speech infront of a big or small audience, facing a stranger,and being the main attraction in any social context.
Pertimbangan semula kaunseling yang berasaskan bantuan kerjaya dalam perkhidmatan kaunselingmembolehkan klien untuk menikmati kesejahteraan psikologi. Sehubungan dengan itu, bantuan kerjaya dianggapsebagai pelbagai perkhidmatan yang dirancangkan oleh seorang kaunselor untuk membantu klien bagimemahami konstruk kerjaya (seperti nilai kerja, konflik dan matlamat) serta pengaruh kontekstual (umpamanya,dasar awam dan fenomena global) yang mempengaruhi kesejahteraan psikologi klien di tempat kerjanya dan didalam keluarga. Umpamanya, bantuan kerjaya yang dirancangkan dengan teliti boleh dilaksanakan sebagaiintervensi yang berbentuk perundingan untuk membantu klien bagi (1) mencantumkan nilai dan matlamat kerja pihak majikan dengan nilai peribadi klien supaya klien boleh menghadapi konflik nilai di tempat kerja; dan (2)menyelaraskan pembangunan kerjaya klien dengan tuntutan pihak majikan supaya klien boleh merasa puas hati.Implikasi kaunseling yang berasaskan bantuan kerjaya dalam konteks Malaysia adalah seorang kaunselor perlu:(1) cekap untuk menjalankan kaunseling kerja; (2) memiliki kecekapan silang budaya; dan (3) mahir untukmelaksana pelbagai intervensi seperti yang diperlukan oleh klien dalam proses kaunseling. Dengan ini, seorangklien dibantu untuk mencapai kesejahteraan psikologi demi menyumbang sumber tenaga bagi transformasinegara ke arah Wawasan 2020.
Citation: Institute for Public Health (IPH) 2012. The National Health and Morbidity Survey: Malaysia Global School-based Student Health Survey 2012. Kuala Lumpur: Ministry of Health Malaysia.
http://iku.moh.gov.my/images/IKU/Document/REPORT/NHMS2012-GlobalSchoolBasedStudentHealthSurvey.pdf
Study name: Global School-based Student Health Survey (GSHS); NHMS-2012
Malaysia School-Based Nutrition Survey 2012
http://iku.moh.gov.my/images/IKU/Document/REPORT/NHMS2012-MalaysiaSchoolBasedNutritionSurvey.pdf
State level reports:
Johor
http://iku.moh.gov.my/images/IKU/Document/REPORT/2012/NHMS2012Johor.pdf
Kedah
http://iku.moh.gov.my/images/IKU/Document/REPORT/2012/NHMS2012Kedah.pdf
Kelantan
http://iku.moh.gov.my/images/IKU/Document/REPORT/2012/NHMS2012Kelantan.pdf
Kuala Lumpur
http://iku.moh.gov.my/images/IKU/Document/REPORT/2012/NHMS2012Kuala%20Lumpur.pdf
Melaka
http://iku.moh.gov.my/images/IKU/Document/REPORT/2012/NHMS2012Melaka.pdf
Negeri Sembilan
http://iku.moh.gov.my/images/IKU/Document/REPORT/2012/NHMS2012NegeriSembilan.pdf
Pahang
http://iku.moh.gov.my/images/IKU/Document/REPORT/2012/NHMS2012Pahang.pdf
Perak
http://iku.moh.gov.my/images/IKU/Document/REPORT/2012/NHMS2012Perak.pdf
Perlis
http://iku.moh.gov.my/images/IKU/Document/REPORT/2012/NHMS2012Perlis.pdf
Pulau Pinang
http://iku.moh.gov.my/images/IKU/Document/REPORT/2012/NHMS2012Penang.pdf
Putrajaya
http://iku.moh.gov.my/images/IKU/Document/REPORT/2012/NHMS2012Putrajaya.pdf
Sabah & Labuan
http://iku.moh.gov.my/images/IKU/Document/REPORT/2012/NHMS2012SabahDanLabuan.pdf
Sarawak
http://iku.moh.gov.my/images/IKU/Document/REPORT/2012/NHMS2012Sarawak.pdf
Selangor
http://iku.moh.gov.my/images/IKU/Document/REPORT/2012/NHMS2012Selangor.pdf
Terengganu
http://iku.moh.gov.my/images/IKU/Document/REPORT/2012/NHMS2012Terengganu.pdf
Citation: ITC Project (March 2012). ITC Malaysia National Report. Findings from Wave 1 to 4 Surveys (2005–2009). University of Waterloo, Waterloo, Ontario, Canada; Universiti Sains Malaysia, Pulau Pinang, Malaysia; and Ministry of Health, Putrajaya, Malaysia
This report presents results of Waves 1 to 4 (2005 – 2009) of the ITC Malaysia Survey – a face-to-face and telephone survey of a cohort of approximately 2,000 adult smokers and, 1,000 youth smokers and non-smokers. Waves 1 to 3 also included a cohort of 1,500 adult non-smokers. The key findings contained in this report provide evidence regarding the attitudes and behaviours of adult and youth smokers and non-smokers to assist policy makers in implementing effective tobacco control policies in Malaysia.
Citation: Global Adult Tobacco Survey (GATS) Malaysia. Kuala Lumpur: Ministry of Health, Malaysia, 2012
Fact sheet: http://moh.gov.my/images/gallery/Report/GATS_Malaysia.pdf
Datasets: GATS_Malaysia_National_2011_SPSS.zip
https://nccd.cdc.gov/GTSSDataSurveyResources/Ancillary/DataReports.aspx?CAID=2
Study name: Global Adults Tobacco Survey (GATS-2011)
MeSH terms: Adult; Cross-Sectional Studies; Health Surveys; Humans; Health Knowledge, Attitudes, Practice; Malaysia; Smoking; Tobacco; Tobacco Smoke Pollution; Prevalence
BACKGROUND: Treatment of hypercholesterolaemia in Asia is rarely evaluated on a large scale, and data on treatment outcome are scarce. The Pan-Asian CEPHEUS study aimed to assess low-density lipoprotein cholesterol (LDL-C) goal attainment among patients on lipid-lowering therapy.
METHODS: This survey was conducted in eight Asian countries. Hypercholesterolaemic patients aged ≥18 years who had been on lipid-lowering treatment for ≥3 months (stable medication for ≥6 weeks) were recruited, and lipid concentrations were measured. Demographic and other clinically relevant information were collected, and the cardiovascular risk of each patient was determined. Definitions and criteria set by the updated 2004 National Cholesterol Education Program guidelines were applied.
RESULTS: In this survey, 501 physicians enrolled 8064 patients, of whom 7281 were included in the final analysis. The mean age was 61.0 years, 44.4% were female, and 85.1% were on statin monotherapy. LDL-C goal attainment was reported in 49.1% of patients overall, including 51.2% of primary and 48.7% of secondary prevention patients, and 36.6% of patients with familial hypercholesterolaemia. The LDL-C goal was attained in 75.4% of moderate risk, 55.4% of high risk, and only 34.9% of very high-risk patients. Goal attainment was directly related to age and inversely related to cardiovascular risk and baseline LDL-C.
CONCLUSION: A large proportion of Asian hypercholesterolaemic patients on lipid-lowering drugs are not at recommended LDL-C levels and remain at risk for cardiovascular disease. Given the proven efficacy of lipid-lowering drugs in the reduction of LDL-C, there is room for further optimization of treatments to maximize benefits and improve outcomes.
MeSH terms: Adult; Hypolipidemic Agents/therapeutic use*; Asia/epidemiology; Cardiovascular Diseases/ethnology; Cardiovascular Diseases/prevention & control; Cross-Sectional Studies; Drug Utilization; Female; Hong Kong; Humans; Hypercholesterolemia/blood; Hypercholesterolemia/drug therapy*; Hypercholesterolemia/ethnology; Indonesia; Korea; Cholesterol, LDL/blood*; Malaysia; Male; Middle Aged; Philippines; Practice Patterns, Physicians'; Drug Prescriptions; Prospective Studies; Risk Factors; Taiwan; Thailand; Time Factors; Vietnam; Biomarkers/blood; Down-Regulation; Multivariate Analysis; Linear Models; Logistic Models; Odds Ratio; Treatment Outcome; Practice Guidelines as Topic; Risk Assessment; Health Care Surveys; Guideline Adherence; Asian Continental Ancestry Group
A 69 year-old man presented with an incidental finding on radiograph of a lesion in the left upper lobe. CT indicated it was likely to be a neoplasm and CT-guided FNA was requested. The lesion was located medial to the scapula so a creative approach was utilised to gain access to the lesion. This study discusses the approach used and why it reduced patient risk compared to a more conventional procedure. The sample was positive for neoplasm and there were no complications arising from the procedure.
Indian spinach (Basella rubra L.) is a red stem species of Basella that is cultivated worldwide as an ornamental and the aerial parts are also consumed as a vegetable. In May of 2011, symptoms of damping-off were observed on approximately 10% of the plants at the stem base around the soil line of seedlings in a greenhouse in Homestead, FL. Lesions were initially water soaked, grayish to dark brown, irregular in shape, and sunken in appearance on large plants, causing the infected seedlings to collapse and eventually die. Symptomatic stem tissue was surface sterilized with 0.6% sodium hypochlorite, rinsed in sterile distilled water, air dried, and plated on potato dextrose agar (PDA). Plates were incubated at 25°C in darkness for 3 to 5 days. A fungus was isolated in all six isolations from symptomatic tissues on PDA. Fungal colonies on PDA were light gray to brown with abundant growth of mycelia, and the hyphae tended to branch at right angles when examined under a microscope. A septum was always present in the branch of hyphae near the originating point and a slight constriction at the branch was observed. Neither conidia nor conidiophores were found from the cultures on PDA. The characteristics of hyphae, especially the right angle branching of mycelia, indicate close similarity to those of Rhizoctonia solani (2,3). The internal transcribed spacer (ITS) region of rDNA was amplified with the primers ITS1/ITS4 and sequenced (GenBank Accession No. JN545836). Subsequent database searches by the BLASTN program indicated that the resulting sequence had a 100% identity over 472 bp with the corresponding gene sequence of R. solani anastomosis group (AG) 4 (GenBank Accession No. JF701752.1), a fungal pathogen reported to cause damping-off on many crops. Pathogenicity was confirmed through inoculation of healthy India spinach plants with the hyphae of isolates. Four 4-week-old plants were inoculated with the isolates by placing a 5-mm PDA plug of mycelia at the stem base and covering with a thin layer of the soil. Another four plants treated with sterile PDA served as a control. After inoculation, the plants were covered with plastic bags for 24 h and maintained in a greenhouse with ambient conditions. Four days after inoculation, water-soaked, brown lesions, identical to the symptoms described above, were observed on the stem base of all inoculated plants, whereas no symptoms developed on the control plants. The fungus was isolated from affected stem samples, and the identity was confirmed by microscopic appearance of the hyphae and sequencing the ITS1/ITS4 intergenic spacer region, fulfilling Koch's postulates. This pathogenicity test was conducted twice. R. solani has been reported to cause damping-off of B. rubra in Ghana (1) and Malaysia (4). To our knowledge, this is the first report of damping-off caused by R. solani AG-4 on Indian spinach in Florida and the United States. With the increased interest in producing Asian vegetables for food and ornamental purposes, the occurrence of damping-off on Indian spinach needs to be taken into account when designing programs for disease management in Florida. References: (1) H. A. Dade. XXIX. Bull. Misc. Inform. 6:205, 1940. (2) J. R. Parmeter et al. Phytopathology 57:218, 1967. (3) B. Sneh et al. Identification of Rhizoctonia species. The American Phytopathological Society, St Paul, MN, 1991. (4) T. H. Williams and P. S. W. Liu. Phytopathol. Pap. 19:1, 1976.
In June 2011, tomatoes (Solanum lycopersicum) in major growing areas of the Cameron Highlands and the Johor state in Malaysia were affected by a leaf spot disease. Disease incidence exceeded 80% in some severely infected regions. Symptoms on 50 observed plants initially appeared on leaves as small, brownish black specks, which later became grayish brown, angular lesions surrounded by a yellow border. As the lesions matured, the affected leaves dried up and became brittle and later developed cracks in the center of the lesions. A survey was performed in these growing areas and 27 isolates of the pathogen were isolated from the tomato leaves on potato carrot agar (PCA). The isolates were purified by the single spore technique and were transferred onto PCA and V8 agar media for conidiophore and conidia production under alternating light (8 hours per day) and darkness (16 hours per day) (4). Colonies on PCA and V8 agar exhibited grey mycelium and numerous conidia were formed at the terminal end of conidiophores. The conidiophores were up to 240 μm long. Conidia were oblong with 2 to 11 transverse and 1 to 6 longitudinal septa and were 24 to 69.6 μm long × 9.6 to 14.4 μm wide. The pathogen was identified as Stemphylium solani on the basis of morphological criteria (2). In addition, DNA was extracted and the internal transcribed spacer region (ITS) was amplified by universal primers ITS5 and ITS4 (1). The PCR product was purified by the commercial PCR purification kit and the purified PCR product sequenced. The resulting sequences were 100% identical to published S. solani sequences (GenBank Accestion Nos. AF203451 and HQ840713). The amplified ITS region was deposited with NCBI GenBank under Accession No. JQ657726. A representative isolate of the pathogen was inoculated on detached 45-day-old tomato leaves of Malaysian cultivar 152177-A for pathogenicity testing. One wounded and two nonwounded leaflets per leaf were used in this experiment. The leaves were wounded by applying pressure to leaf blades with the serrated edge of a forceps. A 20-μl drop of conidial suspension containing 105 conidia/ml was used to inoculate these leaves (3). The inoculated leaves were placed on moist filter paper in petri dishes and incubated for 48 h at 25°C. Control leaves were inoculated with sterilized distilled water. After 7 days, typical symptoms for S. solani similar to those observed in the farmers' fields developed on both wounded and nonwounded inoculated leaves, but not on noninoculated controls, and S. solani was consistently reisolated. To our knowledge, this is the first report of S. solani causing gray leaf spot of tomato in Malaysia. References: (1) M. P. S. Camara et al. Mycologia 94:660, 2002. (2) B. S. Kim et al. Plant Pathol. J. 15:348, 1999. (3) B. M. Pryor and T. J. Michailides. Phytopathology 92:406, 2002. (4) E. G. Simmons. CBS Biodiversity Series 6:775, 2007.
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.
Symptoms of gray leaf spot were first observed in June 2011 on pepper (Capsicum annuum) plants cultivated in the Cameron Highlands and Johor State, the two main regions of pepper production in Malaysia (about 1,000 ha). Disease incidence exceeded 70% in severely infected fields and greenhouses. Symptoms initially appeared as tiny (average 1.3 mm in diameter), round, orange-brown spots on the leaves, with the center of each spot turning gray to white as the disease developed, and the margin of each spot remaining dark brown. A fungus was isolated consistently from the lesions using sections of symptomatic leaf tissue surface-sterilized in 1% NaOCl for 2 min, rinsed in sterile water, dried, and plated onto PDA and V8 agar media (3). After 7 days, the fungal colonies were gray, dematiaceous conidia had formed at the end of long conidiophores (19.2 to 33.6 × 12.0 to 21.6 μm), and the conidia typically had two to six transverse and one to four longitudinal septa. Fifteen isolates were identified as Stemphylium solani on the basis of morphological criteria described by Kim et al. (3). The universal primers ITS5 and ITS4 were used to amplify the internal transcribed spacer region (ITS1, 5.8, and ITS2) of ribosomal DNA (rDNA) of a representative isolate (2). A 570 bp fragment was amplified, purified, sequenced, and identified as S. solani using a BLAST search with 100% identity to the published ITS sequence of an S. solani isolate in GenBank (1). The sequence was deposited in GenBank (Accession No. JQ736024). Pathogenicity of the fungal isolate was tested by inoculating healthy pepper leaves of cv. 152177-A. A 20-μl drop of conidial suspension (105 spores/ml) was used to inoculate each of four detached, 45-day-old pepper leaves placed on moist filter papers in petri dishes (4). Four control leaves were inoculated similarly with sterilized, distilled water. The leaves were incubated at 25°C at 95% relative humidity for 7 days. Gray leaf spot symptoms similar to those observed on the original pepper plants began to develop on leaves inoculated with the fungus after 3 days, and S. solani was consistently reisolated from the leaves. Control leaves did not develop symptoms and the fungus was not reisolated from these leaves. Pathogenicity testing was repeated with the same results. To our knowledge, this is the first report of S. solani causing gray leaf spot on pepper in Malaysia. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) M. P. S. Camara et al. Mycologia 94:660, 2002. (3) B. S. Kim et al. Plant Pathol. J. 15:348, 1999. (4) B. M. Pryor and T. J. Michailides. Phytopathology 92:406, 2002.
A DNA macroarray was previously developed to detect major fungal and oomycete pathogens of solanaceous crops. To provide a convenient alternative for researchers with no access to X-ray film-developing facilities, specific CCD cameras or Chemidoc XRS systems, a chromogenic detection method with sensitivity comparable with chemiluminescent detection, has been developed. A fungal (Stemphylium solani) and an oomycete (Phytophthora capsici) pathogen were used to develop the protocol using digoxigenin (DIG)-labeled targets. The internal transcribed spacer (ITS) region of the nuclear ribosomal DNA (rDNA), including ITS1, 5.8S rDNA, and ITS2, was used as the target gene and polymerase chain reaction amplified as in the previous protocol. Various amounts of species-specific oligonucleotides on the array, quantities of DIG-labeled ITS amplicon, and hybridization temperatures were tested. The optimal conditions for hybridization were 55°C for 2 h using at least 10 pmol of each species-specific oligonucleotide and labeled target at 10 ng/ml of hybridization buffer. Incubation of the hybridized array with anti-DIG conjugated alkaline phosphatase substrates, NBT/BCIP, produced visible target signals between 1 and 3 h compared with 1 h in chemiluminescent detection. Samples from pure cultures, soil, and artificially inoculated plants were also used to compare the detection using chemiluminescent and chromogenic methods. Chromogenic detection was shown to yield similar results compared with chemiluminescent detection in regard to signal specificity, duration of hybridization between the array and targets, and cost, though it takes 1 to 2 h longer for the visualization process, thus providing a convenient alternative for researchers who lack darkroom facilities. To our knowledge, this is the first report of DNA macroarray detection of plant pathogens using a chromogenic method.
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.
Physic nut (Jatropha curcas L.) is an important biofuel crop worldwide. Although it has been reported to be resistant to pests and diseases (1), stem cankers have been observed on this plant at several locations in Peninsular Malaysia since early February 2008. Necrotic lesions on branches appear as scars with vascular discoloration in the tissue below the lesion. The affected area is brownish and sunken in appearance. Disease incidence of these symptomatic nonwoody plants can reach up to 80% in a plantation. Forty-eight samples of symptomatic branches collected from six locations (University Farm, Setiu, Gemenceh, Pulau Carey, Port Dickson, and Kuala Selangor) were surface sterilized in 10% bleach, rinsed twice with sterile distilled water, air dried on filter paper, and plated on water agar. After 4 days, fungal colonies on the agar were transferred to potato dextrose agar (PDA) and incubated at 25°C. Twenty-seven single-spore fungal cultures obtained from all locations produced white, aerial mycelium that became dull gray after a week in culture. Pycnidia from 30-day-old pure cultures produced dark brown, oval conidia that were two celled, thin walled, and oval shape with longitudinal striations. The average size of the conidia was 23.63 × 12.72 μm with a length/width ratio of 1.86. On the basis of conidial morphology, these cultures were identified as Lasiodiplodia theobromae. To confirm the identity of the isolates, the internal transcribed spacer (ITS) region was amplified with ITS1/ITS4 primers and sequenced. The sequences were deposited in GenBank (Accession Nos. HM466951, HM466953, HM466957, GU228527, HM466959, and GU219983). Sequences from the 27 isolates were 99 to 100% identical to two L. theobromae accessions in GenBank (Nos. HM008598 and HM999905). Hence, both morphological and molecular characteristics confirmed the isolates as L. theobromae. Pathogenicity tests were performed in the glasshouse with 2-month-old J. curcas seedlings. Each plant was wound inoculated by removing the bark on a branch to a depth of 2 mm with a 10-mm cork borer. Inoculation was conducted by inserting a 10-mm-diameter PDA plug of mycelium into the wound and wrapping the inoculation site with wetted, cotton wool and Parafilm. Control plants were treated with plugs of sterile PDA. Each isolate had four replicates and two controls. After 6 days of incubation, all inoculated plants produced sunken, necrotic lesions with vascular discoloration. Leaves were wilted and yellow above the point of inoculation on branches. The control plants remained symptomless. The pathogen was successfully reisolated from lesions on inoculated branches. L. theobromae has been reported to cause cankers and dieback in a wide range of hosts and is common in tropical and subtropical regions of the world (2,3). To our knowledge, this is the first report of stem canker associated with L. theobromae on J. curcas in Malaysia. References: (1) S. Chitra and S. K. Dhyani. Curr. Sci. 91:162, 2006. (2) S. Mohali et al. For. Pathol. 35:385, 2005. (3) E. Punithalingam. Page 519 in: CMI Descriptions of Pathogenic Fungi and Bacteria. Commonwealth Mycological Institute, Kew, Surrey, UK. 1976.