Displaying publications 61 - 80 of 255 in total

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  1. Siti Aisyah Abd Ghafar, Muhammad Fikhry Mohd Salehuddin,, Nur Syamimi Syuhada Che Awang, Rohazila Mohamad Hanafiah
    MyJurnal
    Introduction:Spilanthes acmella, also known as “subang nenek’, has been used traditionally in Malaysia to treat toothache. A previous study has shown Spilanthes acmella leaves extracts (SALE) inhibit Streptococcus mutans growth. Streptococcus mutans is commonly found in the human oral cavity and is the main contributor to tooth de-cay. There is no study on the antibacterial effects of Spilanthes acmella flower extracts (SAFE) against Streptococcus mutans reported to date. Therefore, the objective of this study is to investigate antibacterial properties of SAFE against S. mutans. Methods:S. mutans was subcultured in Muller Hinton (MH) broth and agar. Sequential extractions of S. acmella flowers were conducted using four different solvents with increasing polarity, [n- hexane, dichloromethane (DCM), acetone, methanol (MeoH)] and tested with different concentrations against S. mutans via the disc diffusion assay, minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC). Sodium fluoride (NaF) was used as a positive control while DMSO was used as a negative control. Results: The disc diffusion assay shows SAFE inhibited Streptococcus mutans growth. SAFE-DCM shows the greatest inhibition properties (12.33±2.30 mm) followed by SAFE-n-hexane (11.33±0.57 mm). Meanwhile, SAFE-Meoh and SAFE-acetone show no inhibition zone (6.00±0.001 mm). MIC value for SAFE-DCM and SAFE-n-hexane is 12.5 mg/mL respectively. Whereas, MBC value SAFE-DCM and SAFE-n-hexane is 50.0 mg/mL respectively. Conclusion: It can be concluded SAFE-DCM and SAFE-n-hexane possesses bactericidal properties against Streptococcus mutans.
    Matched MeSH terms: Agar
  2. Vankayala B, Anantula K, Saladi H, Gudugunta L, Basavarajaiah JM, Yadav SS
    J Conserv Dent, 2020 08 20;22(6):559-563.
    PMID: 33088065 DOI: 10.4103/JCD.JCD_221_19
    Aim: This study aims to evaluate the amount of apical extrusion of bacteria during root canal instrumentation using K3XF, Protaper Gold, Edge taper platinum, and Hyflex CM Rotary systems.

    Materials and Methods: Sixty freshly extracted maxillary incisors teeth collected in saline. Access cavity prepared and canals were made free of bacterial and pulp. The teeth were mounted on the bacteria collecting apparatus. Root canals were contaminated with the Fusobacterium Nucleatum (ATCC25586) and dried at 37°C for 24 h. In Group 1 (Control group): No instrumentation was done and biomechanical preparation done in all other groups with Group 2: Hand K-files, Group 3: Protaper gold, Group 4: K3XF, Group 5: Edge taper platinum, and Group 6: Hyflex CM rotary file systems. Then, the extrude was collected, and it is incubated in Mueller-Hinton agar for 24 h and the number of colony forming units were counted and statistical comparison was done using Kruskal-Wallis test and Mann-Whitney U test.

    Results: Hand K-files extruded more bacteria when compared to other four rotary systems, K3XF file system extruded least number of bacteria.

    Conclusion: All instrumentation techniques extruded intracanal bacteria apically. However, engine-driven nickel-titanium instruments extruded less bacteria than the manual technique. The K3XF rotary file system comparatively extruded less bacteria than other rotary file systems.

    Matched MeSH terms: Agar
  3. Farra Amira Mohamed, Aimi Nadia Ramli,, Noorlis Ahmad
    MyJurnal
    Demand for milk has increased in Malaysia due to the increased in awareness of healthy foods consumption.
    Hence, research of milk is crucial to ensure that it is not contaminated with Escherichia coli. This study
    evaluated the survival of Escherichia coli at different temperature and haemolysin activity of Escherichia
    coli on blood agar. A total of 8 samples of raw fresh and pasteurized milk were collected from nearby farm
    and market in Negeri Sembilan, Malaysia. After an overnight exposure to four different temperatures of
    0
    0C, 280C, 350C and 450C, the bacteriological test of milk was evaluated for the presence of Escherichia
    coli. Overall, all raw fresh milk sampled exceeded the acceptable limit of bacterial count of 1 x 105 CFU/ml.
    Raw fresh milk recorded the highest count at 35oC with 4.4 x 107 CFU/ml and the lowest at 0oC with 8.3 x
    104 CFU/ml. The presence of Escherichia coli was detected in 7/20(35%) of the total raw fresh milk
    samples. All pasteurized milk showed no presence of Escherichia coli due to the effectiveness of heat
    treatment. Haemolysin test showed no haemolytic activity. Milk contaminated with Escherichia coli can
    cause diarrheal, gastrointestinal diseases and urinary infection. Hence, it is important to study the survival
    rate of Escherichia coli and its pathogenicity in milk to ensure public safety.
    Matched MeSH terms: Agar
  4. Nur Hafizah Zakaria, Husnul Azan Tajarudin, Mohd Sharizal Mohd Sapingi, Mohamad Fared Murshed
    Scientific Research Journal, 2017;14(1):42-52.
    MyJurnal
    This study focused on the identification of pathogenic bacteria in raw water intake and after sand filtration for drinking water treatment plant during flood event in 2014. The samples was collected from the Lubok Buntar Water Treatment Plant (WTP) and processed through bacterial isolation using chocolate agar as a media. The isolation process conducted based on serial samples dilution and streaking method prior to DNA extraction. Deoxyribonucleic acid (DNA) extraction kit was used to get selected bacteria DNA and further analysis using Polymerase Chain Reaction (PCR) test and electrophoresis to get DNA sequences. The Basic Local Alignment Search Tool (BLAST) analysis was employed to identify the species of the isolated bacteria. As a result, Pantoeaagglomerans and Enterobacter sp. were found in raw and filtered water sample and indicating the same family types. It was concluded that bacteria of the same species were found before and after sand filtration and need to be removed by disinfectant process. The findings also indicated that all the physicochemical parameters measured were within the values prescribed by the Interim National Water Quality Standard (INWQS).
    Matched MeSH terms: Agar
  5. Heng PY, Sulong A, Ali UKS, Wong KK
    Malays J Pathol, 2019 Aug;41(2):139-148.
    PMID: 31427549
    INTRODUCTION: OXA-48, a carbapenem-hydrolysing class D β-lactamase, and its variant, OXA-181, are increasingly reported worldwide. This study aimed to describe the prevalence and distribution of OXA-48 and OXA-181 carbapenem-resistant Enterobacteriaceae (CRE) in a tertiary medical centre in Malaysia.

    MATERIALS & METHODS: A total of 13,098 Enterobacteriaceae isolates from various clinical samples were sent to our laboratory between January 2011 and December 2012. Of these, 90 demonstrated reduced susceptibility to at least one carbapenem and were included in this study. Only 88 isolates were successfully subcultured on blood agar (BA). Another 2 isolates failed to grow and were excluded. Of the 88, 2 isolates had the same identification number (repetitive isolates); therefore, 1 isolate was excluded from further analyses. Only 87 isolates were subjected to molecular detection of the blaOXA-48 and blaOXA-181 genes by polymerase chain reaction.

    RESULTS: Eighty-seven non-repetitive isolates grew following subculture on BA. Of these, 9 (10.34%) were positive for OXA-48 (7 Klebsiella pneumoniae, 2 Escherichia coli). Each isolate originated from different patients. All patients had a history of treatment with at least one cephalosporin and/or carbapenem prior to the isolation of OXA-48 CRE. OXA-181 was detected in one (1.15%) out of the 87 isolates; CONCLUSIONS: The prevalence of OXA-48 and OXA-181 CRE among all Enterobacteriaceae isolates in our institution is 0.069% and 0.008%, respectively. Nevertheless, our findings suggest that OXA-48 and OXA-181 carbapenemases appear to be important and possibly under-recognised causes of carbapenem resistance in Malaysia.

    Matched MeSH terms: Agar
  6. Khalid A, Ahmad P, Alharthi AI, Muhammad S, Khandaker MU, Rehman M, et al.
    Nanomaterials (Basel), 2021 Feb 10;11(2).
    PMID: 33578945 DOI: 10.3390/nano11020451
    Copper oxide and Zinc (Zn)-doped Copper oxide nanostructures (CuO-NSs) are successfully synthesized by using a hydrothermal technique. The as-obtained pure and Zn-doped CuO-NSs were tested to study the effect of doping in CuO on structural, optical, and antibacterial properties. The band gap of the nanostructures is calculated by using the Tauc plot. Our results have shown that the band gap of CuO reduces with the addition of Zinc. Optimization of processing conditions and concentration of precursors leads to the formation of pine needles and sea urchin-like nanostructures. The antibacterial properties of obtained Zn-doped CuO-NSs are observed against Gram-negative (Pseudomonasaeruginosa,Klebsiellapneumonia,Escherichiacoli) and Gram-positive (Staphylococcusaureus) bacteria via the agar well diffusion method. Zn doped s are found to have more effective bacterial resistance than pure CuO. The improved antibacterial activity is attributed to the reactive oxygen species (ROS) generation.
    Matched MeSH terms: Agar
  7. Ong AH, Vellayan S
    Zoo Biol, 2008 Jan;27(1):62-9.
    PMID: 19360604 DOI: 10.1002/zoo.20163
    The amplification of the highly conserved chromo-helicase-DNA binding region found in both the Z and W chromosome was evaluated with three sets of primers (P8/P2, 1237L/1272H and 2550F/2718R). DNA extracted from feathers through a simple boiling method was used to address its reliability in generating the sex-linked bands. All the bird samples, including the seven bird families that have not been reported previously, were successfully amplified with the primer set 2550F/2718R. The resulting polymerase chain reaction products showed clearly resolved fragments on a conventional agarose gel electrophoresis with size differences ranging from 80 to 540 bp between the two respective ZW gene copies. Although the P8/P2 primer was not as effective under the same conditions, it was able to produce well-resolved Z and W bands from bird species under the Antidea family, whereas the 2250F/2718R primer set only produced a single amplified fragment of a different size between the male and the female. Zoo Biol 27:62-69, 2008. (c) 2007 Wiley-Liss, Inc.
    Matched MeSH terms: Electrophoresis, Agar Gel
  8. Nur Azura AB, Yusoff M, Tan GY, Jegadeesh R, Appleton DR, Vikineswary S
    J Ind Microbiol Biotechnol, 2016 Apr;43(4):485-93.
    PMID: 26721619 DOI: 10.1007/s10295-015-1724-4
    Actinomycete strain AUM 00500 was 99.5 % similar to Streptomyces sanglieri NBRC 100784(T) and was evaluated for antagonistic activity towards Ganoderma boninense, the causative fungus of basal stem rot of oil palm. The strain showed strong antifungal activity towards G. boninense in in vitro and SEM analysis showed various modes of inhibition of the fungus. Ethyl acetate extracts of single culture and inhibition zone of cross-plug culture by HPLC indicated that strain AUM 00500 produced two different antibiotics of the glutarimide group namely cycloheximide and actiphenol. In greenhouse trials, oil palm seed treated with spores of S. sanglieri strain AUM 00500 at 10(9) cfu/ml showed significant (P agar.
    Matched MeSH terms: Agar
  9. Ong CC, Yusoff K, Yap CK, Tan SG
    J Genet, 2009 Aug;88(2):153-63.
    PMID: 19700853
    A total of 19 polymorphic microsatellite loci were used to analyse levels of genetic variation for 10 populations of Perna viridis L. collected from all over peninsular Malaysia. The populations involved in this study included Pulau Aman in Penang, Tanjung Rhu in Kedah, Bagan Tiang in Perak, Pulau Ketam in Selangor, Muar, Parit Jawa, Pantai Lido and Kampung Pasir Puteh in Johore, and Kuala Pontian and Nenasi in Pahang state. The number of alleles per locus ranged from two to seven, with an average of 3.1. Heterozygote deficiencies were observed across all the 10 populations. Characterization of the populations revealed that local populations of P. viridis in peninsular Malaysia were genetically similar enough to be used as a biomonitoring agent for heavy metal contamination in the Straits of Malacca. Cluster analysis grouped the P. viridis populations according to their geographical distributions with the exception of Parit Jawa. The analysis also revealed that P. viridis from the northern parts of peninsular Malaysia were found to be the most distant populations among the populations of mussels investigated and P. viridis from the eastern part of peninsular Malaysia were closer to the central and southern populations than to the northern populations.
    Matched MeSH terms: Electrophoresis, Agar Gel
  10. Teoh, Chul Peng, Koh, Soon Peng, Clemente Michael Wong Vui Ling
    MyJurnal
    Glaciozyma antarctica PI12 is a psychrophilic yeast isolated from Antarctica. It has an optimal growth in yeast peptone dextrose (YPD) and yeast mould (YM) broth media but not in potato dextrose (PD) broth medium. Early phase G. antarctica PI12 cells had elongated-shape and became oval-shaped as they aged. G. antarctica PI12 exhibited bipolar budding and formed a chain of cells during the lag and early exponential phases. The number of chains decreased as the yeast aged. It appeared mainly as a single cell at the stationary phase, and a small number of them still produced buds. Some cells at the stationary phase entered the quiescence state (G0) as a longterm survival strategy. The G. antarctica PI12 cell size decreased when they entered the stationary phase. G. antarctica PI12 was found to produce hydrolytic enzymes, chitinase, cellulase, mannanase, and xylanase. A higher glucose concentration of 2% in the PD agar medium inhibited the activities of chitinase but not the cellulase, mananase and xylanase.
    Matched MeSH terms: Agar
  11. Omar S, Jalaludin FA, Yee JM, Kamarudin Z, Jayaseelan K, Khlubi ANM, et al.
    J Vet Med Sci, 2020 Aug 28;82(8):1236-1242.
    PMID: 32641623 DOI: 10.1292/jvms.20-0229
    It is important to provide a baseline of fungal composition in the captive wildlife environment to better understand their role in overall wildlife health. The objectives were to identify species of fungi existing within wildlife animal enclosures and their environment at the National Wildlife Rescue Centre (NWRC) and the National Zoo, Malaysia and to describe their medical and veterinary importance. Samples of air, wall or floor swab, enrichment swab and soil were taken from the animal enclosures, exercise yard and enrichments at NWRC and National Zoo respectively. All samples including those pre-treated samples were plated onto Sabouraud's Dextrose Agar (SDA). Numerous fungi were grown on all sampling SDA plates regardless by either single or multiple growth. Samples of air in both NWRC and National Zoo had the highest growth of Penicillium spp. with a prevalence of 31.2% and 83.7% respectively. Samples of swab from the wall, floor and enrichments were predominantly by Candida spp. (42.6%) in NWRC and Penicillium spp. (41.6%) in the National Zoo. Prevalence of multiple fungi isolated from the soil samples in NWRC were 57.9% and yeast species was the most common in National Zoo with a prevalence of 88.9%. Overall, 29 and 8 isolates were found in both samples from the NWRC and National Zoo with a predominant species of potential zoonotic fungi have been identified in both premises. The expected fungus Aspergillus spp. was not isolated in all samples in NWRC. Prevalent fungal species found in this study are known to cause disease in animals and humans as primary pathogen and also as opportunistic pathogens that may also cause infection. Thus, health safety precautions should be considered particularly in dealing with conservation of endangered wildlife species, along with personnel and public involvements.
    Matched MeSH terms: Agar
  12. Jibril FI, Mohd Hilmi AB, Aliyu S
    J Pharm Bioallied Sci, 2020 Nov;12(Suppl 2):S831-S835.
    PMID: 33828385 DOI: 10.4103/jpbs.JPBS_280_19
    Introduction: Stingless bee is an insect that belongs to the family Apidae. Its name is based on its disability of stinging. It has a high product of Meliponini honey and propolis by which are commonly referred to as stingless bee honey and stingless bee propolis. Meliponini honey is one of the crucial natural sources and has the potential to kill infectious microorganisms. Previous studies have proved that the antibacterial activity of natural honey was an effect of hydrogen peroxide, a substance contained in the honey. However, these claims were contradicting with too many studies.

    Objective: Therefore, this study aimed to identify the antibacterial activity of Malaysian Meliponini honey which contained non-hydrogen peroxide against Staphylococcus aureus, an opportunistic microbial.

    Materials and Methods: Meliponini honey was used as an antibacterial agent for the treatment of S. aureus in agar well diffusion assay. An amplex red hydrogen peroxide kit was used to identify the hydrogen peroxide in the honey sample. Meanwhile, non-hydrogen peroxide activity was performed by using honey-catalase treated.

    Results: For the first time, we found that hydrogen peroxide was absent in all Meliponini honey samples. Meliponini honey has higher antibacterial activity (13.30 ± 0.56mm) compared to Apis honey (9.03 ± 0.22mm) in agar well diffusion assay.

    Discussion: Non-hydrogen peroxide in Meliponini honey is a bioactive compound and beneficial to kill the microbial infection.

    Conclusion: Antibacterial activity of Malaysian Meliponini honey is directly contributed by non-hydrogen peroxide.

    Matched MeSH terms: Agar
  13. Cheung, Tian Pei, Rostenberghe, Hans Van, Narazah Mohd Yusoff, Noraida Ramli, Nor Rosidah Ibrahim, Nishio, Hisahide, et al.
    MyJurnal
    Background: The low yield and quality of buccal-derived genomic DNA have reduced its applicability in various genetic research. The aim of this study was to assess the quantity, purity and genotyping efficiency of genomic DNA isolated from neonatal buccal swabs. Methods: Paired buccal swabs and whole blood samples were collected from 60 neonates with the mean age 5 days (SD=1.57). The genomic DNA quantity and purity were measured by using Infinite® 200 PRO NanoQuant reader and agarose gel electrophoresis. High-resolution melting (HRM) analysis was used to analyse the sequence variants present in uridine diphosphate glucuronosyltransferase 1A1 (UGT1A1 c.211G>A) and nuclear receptor subfamily 1, group I, member 3 (NR1I3 IVS8+116T>G) genes. Results: Buccal swabs provided lower mean genomic DNA concentration (18.78 ± 8.39 ng/μl versus 40.02 ± 13.03 ng/μl), yield (2.63 ± 1.17 μgversus8.00 ± 2.61 μg). The purity of buccal samples however were inconsistent with 16 samples (26.7%) having A260/280 ratios below 1.8 which indicated protein contamination. Genomic DNA purity for all blood samples were within the ideal range with average absorbance ratios of 1.8−2.0. However, all buccal genomic DNA demonstrated 100% genotype call rates for all variants. A complete genotype concordance was also observed between paired genomic DNA samples. Conclusion: Despite related to a reduced quantity and purity, neonatal buccal genomic DNA could generate reliable HRM genotyping results. Therefore, buccal swab collection is a promising alternative to the invasive blood sampling to provide genomic DNA for genetic analysis involving paediatric population.
    Matched MeSH terms: Electrophoresis, Agar Gel
  14. Nurul Fatihah Mohamed Yusoff, Basma Ezzat Mustafa, Pram Kumar Subramaniam, Nazih Shaban Mustafa, Muhannad Ali Kashmoola, Khairani Idah Mokhtar, et al.
    MyJurnal
    Introduction:Linum usitatissimum (flax seed) has been cultivated for domestic use since prehistoric times. Its use as a dietary supplement becomes more popular nowadays. Nigella sativa seeds and oils have been widely used for centuries in the treatment of various ailments throughout the world. It is an important drug in the Indian traditional system of medicine like Unani and Ayurveda. Methods: This is a laboratory experimental in-vitro study using select-ed oral pathogens (Streptococcus mutans, Klebsiella pneumoniae and Pseudomonas aeruginosa) cultured in nutrient agar. The pathogens were then inoculated in nutrient based broth and incubation for 24hours. Linum usitatissimum and Nigella sativa extract efficacy was tested by measurement of the zone of inhibition. The result of the extracts antimicrobial activities were compared with positive control (penicillin) and negative control(Dimethyl sulfoxide DMSO). The statistical analysis was done by using SPSS18. Results: The antibacterial effect of Linum usitatissimum and Nigella sativa extract is comparable to the effect of penicillin and this study shows that flax seed extract shows more potent antibacterial effect than Nigella sativa on Streptococcus mutans and Pseudomonas aeruginosa while both extracts didn’t show an effect on Klebsiella pneumoniae. Conclusion: The results of the present study scien-tifically validate the inhibitory capacity of Linum usitatissimum or Nigella sativa as antibiotic against selective oral pathogens this will contribute towards the development of new treatment options based on natural base products.
    Matched MeSH terms: Agar
  15. Singh S, Srivastava B, Gupta K, Gupta N, Singh R, Singh S
    Int J Clin Pediatr Dent, 2020 8 4;13(2):119-123.
    PMID: 32742086 DOI: 10.5005/jp-journals-10005-1718
    Aim and objective: The aim of this study was to evaluate and compare the antifungal efficacy of MTA Fillapex, Metapex, zinc oxide eugenol cement, Endomethasone, and Endoflas against Candida albicans.

    Materials and methods: Root canal exudates of 30 patients were tested against MTA Fillapex (Angelus), Metapex (BioMed), zinc oxide eugenol (Deepak Enterprise), Endomethasone (Septodont), Endoflas FS (Sanlor Laboratories), MTA (Angelus) (positive control), and glycerine (negative control). Children with failed endodontic cases were included in the study. Tube dilution and agar diffusion methods were used to check the antifungal efficacy of the root canal sealers. In tube dilution method, 24-well culture plates containing freshly mixed material along with Candida albicans were used. Wells containing MTA (Angelus) along with Sabouraud dextrose agar and Candida albicans served as positive control while glycerine along with Sabouraud dextrose agar and Candida albicans served as negative control. All plates were incubated at 37°C for 24 hours. Growth of the fungi was monitored after 24 hours by the presence of the turbidity. The samples were recultured to test the experimental material using agar well diffusion method, and the Petri plates were incubated for 24 hours and 72 hours. Zone of inhibition was measured after respective time period. Paired t test was used for the data analysis.

    Results: It was seen in tube dilution method Endomethasone showed least turbidity while maximum was shown by Metapex; similar results were seen in case of agar well diffusion method in which largest zone of inhibition was shown by Endomethasone while smallest was by Metapex.

    Conclusion: It was concluded that Endomethasone showed maximum efficacy against Candida albicans as compared to Metapex.

    How to cite this article: Singh S, Srivastava B, Gupta K, et al. Comparative Evaluation of Antifungal Efficacy of Five Root Canal Sealers against Clinical Isolates of Candida albicans: A Microbiological Study. Int J Clin Pediatr Dent 2020;13(2):119-123.

    Matched MeSH terms: Agar
  16. Harika K, Shenoy VP, Narasimhaswamy N, Chawla K
    J Glob Infect Dis, 2020 08 29;12(3):129-134.
    PMID: 33343163 DOI: 10.4103/jgid.jgid_150_19
    Background: Microorganisms are known to be involved in the formation of biofilm. These biofilms are often seen in chronic wound infections, surgical site infections, implants etc., These are capable of causing recalcitrant infections and most of them are also known to possess high antibiotic resistance.

    Objectives: This study was conducted to detect the biofilm formation in bacterial isolates from chronic wound infections.

    Materials and Methods: In the present study, ninety two isolates from chronic wound infections were identified by MALDI-TOF-MS (bioMerieux) and VITEK-2-MS (bioMerieux). These isolates were further screened for biofilm formation by three methods i. e., Tissue Culture Plate method (TCP), Tube Method (TM) and Congo Red Agar (CRA) method. Impact of biofilm production was correlated with the antibiotic resistant pattern.

    Statistical Analysis: Statistical analysis was done for all three methods considering TCP as Gold Standard and parameters like senitivity and specificity of TM i.e. 47.2 and 100% respectively.

    Results: Out of 92 isolates, biofilm formation was seen in 72 isolates (78.2%) by TCP method. 64 isolates were strong biofilm producers, 8 isolates were moderate biofilm producers and 20 isolates were nonbiofilm producing. High prevalence of biofilm formation was seen in nonhealing ulcers infected with Staphylococcus aureus followed by Klebsiella pneumoniae.

    Conclusion: Among three screening methods used for detection of biofilm production, TCP method is considered to be a standard and most reliable for screening of biofilm formation in comparison to TM and CRA.

    Matched MeSH terms: Agar
  17. Nazerian E, Sijam K, Zainal Abidin MA, Vadamalai G
    Plant Dis, 2011 Nov;95(11):1474.
    PMID: 30731752 DOI: 10.1094/PDIS-10-10-0754
    Cucumber (Cucumis sativus L.) is one of the most important vegetable fruits in Malaysia. Cucumber is principally grown in the states of Johor, Kelantan, and Perak. The broad host range Enterobacteriaceae pathogen, Pectobacterium carotovorum, can cause soft rot on stems or cucumber fruit. In Malaysia, cucumber is produced in a warm, humid climate, thus the plant is susceptible to attack by P. carotovorum at any time during production. In 2010, cucumber samples with wilted and chlorotic leaves, water-soaked lesions, and collapsed fruits were found in multiple fields. Small pieces of infected stems and fruit were immersed in 5 ml of saline solution (0.85% NaCl) for 20 min and then 50 μl of this suspension was spread onto nutrient agar (NA) and incubated at 27°C for 24 h. White-to-pale gray colonies with irregular margins were selected for analysis. For pathogenicity tests, cucumber fruits were surface sterilized by ethyl alcohol 70%, washed with sterilized distilled water, cut into small pieces, and inoculated with 20 μl of 108 CFU/ml suspensions of five representative strains. Cucumber plants were grown for 3 weeks in sterilized soil and their stems were inoculated with 20 μl of 108 CFU/ml of bacterial suspension. Inoculated samples and control (noninoculated) plants were placed in a growth chamber with 80 to 90% relative humidity at 27°C. Symptoms occurred on fruit slices and stems after 1 to 3 days and appeared the same as naturally infected samples, but the control samples remained healthy. Koch's postulates were fulfilled with the reisolation of cultures with the same characteristics as described earlier. Hypersensitivity reaction (HR) assays were done by infiltrating 108 CFU/ml of bacterial suspension into tobacco leaf epidermis and HR developed. All strains were subjected to biochemical and morphological assays, as well as molecular assessment. The strains were gram negative, facultative anaerobes, rod shaped, able to macerate potato slices and growth at 37°C; catalase positive; oxidase and phosphatase negative; able to degrade pectate; sensitive to erythromycin; negative for utilization of α-methyl glycoside, indole production, and reduction of sugars from sucrose; acid production from arabitol, sorbitol, and utilization of citrate were negative, but positive for raffinose and melibiose utilization. PCR amplification of the pel gene by Y1 and Y2 primers produced a 434-bp fragment on agarose gel 1% (1). Amplification of intergenic transcribed spacer region by G1 and L1 primers gave two main bands at approximately 535 and 580 bp on agarose gel 1.5%. The ITS-PCR products were digested with RsaI restriction enzyme (3). On the basis of biochemical and morphological characteristics, PCR-based pel gene and characterization of the ITS region, and digestion of the ITS-PCR products with RsaI restriction enzyme, all isolates were identified as P. carotovorum subsp. carotovorum. To our knowledge, this is the first report of soft rot caused by P. carotovorum subsp. carotovorum on cucumber from Malaysia. References: (1) A. Darraas et al. Appl. Environ. Microbiol. 60:1437, 1994. (2) N. W Schaad et al. Laboratory Guide for the Identification of Plant Pathogenic Bacteria. 3rd ed. The American Phytopathological Society Press, St. Paul, 2001. (3) I. K. Toth et al. Appl. Environ. Microbiol. 67:4070, 2001.
    Matched MeSH terms: Agar
  18. Rooney-Latham S, Blomquist CL, Scheck HJ
    Plant Dis, 2011 Nov;95(11):1478.
    PMID: 30731749 DOI: 10.1094/PDIS-03-11-0261
    Passiflora edulis Sims f. edulis, known as purple passion fruit, is a woody, perennial vine that is grown for its attractive two-part flower and its purple, edible fruit (4). In November 2009, passion fruit vines were collected during a regulatory nursery inspection in Santa Barbara County and submitted to the California Department of Food and Agriculture Plant Pest Diagnostics Laboratory. Nearly 100% of the plants inspected, all of which were approximately 1.25 m tall, appeared stunted, defoliated, and severely wilted. Dark brown vascular discoloration was present in the roots and lower stems of the plants. A pinkish violet Fusarium oxysporum colony containing chlamydospores, multiseptate macroconidia, and microconidia formed on monophialidic conidiophores was consistently isolated from roots and stems onto half-strength acidified potato dextrose agar (aPDA). All further experiments were done with an isolate obtained from a single conidium. A portion of the translation elongation factor gene (TEF-1α) was amplified and sequenced with primers ef1 and ef2 from our isolate (GenBank No. JF332039) (3). BLAST analysis of the 615-bp amplicon with the FUSARIUM-ID database showed 99% similarity with a F. oxysporum passion fruit isolate from Australia (NRRL 38273) (3). To confirm pathogenicity, washed roots of four-leaf stage seedlings approximately 10 cm tall were submerged in a conidial spore suspension (106 spores/ml) for 15 min. The conidial suspension was prepared by flooding 10-day-old cultures grown on aPDA medium with sterile distilled water. Seven seedlings were inoculated and planted in 10-cm2 pots and kept in a 25°C growth chamber with a 12-h photoperiod. Seven seedlings were mock inoculated with sterile water. After 3 weeks, four of the seven inoculated plants had leaves with yellow veins and discolored roots and had partially defoliated. Two of the four symptomatic plants also had brown stem cankers. F. oxysporum grew from the isolated roots and stems of all the inoculated plants. F. oxysporum did not grow from root and stem pieces from the water-dipped plants and the plants remained asymptomatic. Inoculations were repeated on plants approximately 15 cm tall with F. oxysporum growing from roots and stem pieces of all inoculated plants. Symptoms of yellow veins and root necrosis were not observed until 4 weeks after inoculation. Fusarium wilt caused by F. oxysporum f. sp. passiflorae is a significant disease of P. edulis f. edulis in Australia. The disease has also been reported in South Africa, Malaysia, Brazil, Panama, and Venezuela; but it is unclear as to whether the symptoms were caused by Fusarium wilt or Haematonectria canker (1). Banana poka (P. mollissima), P. ligularis, and P. foetida are also susceptible hosts (2). To our knowledge, this is the first report of Fusarium wilt caused by F. oxysporum f. sp. passiflorae on passion fruit in North America. Passion fruit is not commercially produced for consumption in California so the economic importance of this disease appears to be limited to nursery production and ornamental landscapes. The grower of the California nursery stated that the infected passion fruit plants had been propagated on site from seed. The source of inoculum at this nursery remains unknown. References: (1) I. H. Fischer and J. A. M. Rezende. Pest Tech. 2:1, 2008 (2) D. E. Garder. Plant. Dis. 73:476, 1989. (3) D. M. Geiser et al. Eur. J. Plant Pathol. 110:473, 2004. (4) F. W. Martin et al. Econ. Bot. 24:333, 1970.
    Matched MeSH terms: Agar
  19. Intan Sakinah MA, Suzianti IV, Latiffah Z
    Plant Dis, 2013 Jul;97(7):991.
    PMID: 30722542 DOI: 10.1094/PDIS-10-12-0985-PDN
    Banana is the second largest cultivated fruit crop in Malaysia, and is cultivated for both the domestic market and also for export. Anthranose is a well-known postharvest disease of banana and with high potential for damaging market value, as infection commonly occurs during storage. Anthracnose symptoms were observed on several varieties of banana such as mas, berangan, awak, nangka, and rastali in the states of Perak and Penang between August and October 2011. Approximately 80% of the fruits became infected with initial symptoms characterized as brown to black spots that later became sunken lesions with orange or salmon-colored conidial masses. Infected tissues (5 × 5 mm) were surface sterilized by dipping in 1% sodium hypochlorite (NaOCl) for 3 to 5 min, rinsed with sterile distilled water, and plated onto potato dextrose agar (PDA). Direct isolation was done by transferring the conidia from conidial masses using an inoculation loop and plating onto PDA. For both methods, the PDA plates were incubated at 27 ± 1°C with cycles of 12 h light and 12 h darkness. Visible growth of mycelium was observed after 4 to 5 days of incubation. Twenty isolates with conidial masses were recovered after 7 days of incubation. The isolates produced grayish white to grayish green and grey to moss dark green colony on PDA, pale orange conidial masses, and fusiform to cylindrical and hyaline conidia with an average size of 15 to 19 × 5 to 6 μm. Appresoria were ovate to obovate, dark brown, and 9 to 15 × 7 to 12 μm and setae were present, slightly swollen at the base, with a tapered apex, and brown. The cultural and morphological characteristics of the isolates were similar to those described for C. gleosporioides (1,2,3). All the C. gloeosporioides isolates were deposited in culture collection at Plant Pathology Lab, University Sains Malaysia. For confirmation of the identity of the isolates, ITS regions were sequenced using ITS4 and ITS5 primers. The isolates were deposited in GenBank with accessions JX163228, JX163231, JX163201, JX163230, JX163215, JX163223, JX163219, JX163202, JX163225, JX163222, JX163206, JX163218, JX163208, JX163209, JX163210, JX431560, JX163212, JX163213, JX431540, and JX431562. The resulting sequences showed 99% to 100% similarity with multiple C. gloeosporioides isolates in GenBank. Pathogenicity tests were conducted using mas, berangan, awak, nangka, and rastali bananas. Fruit surfaces were sterilized with 70% ethanol and wounded using a sterile scalpel. Two inoculation techniques were performed separately: mycelia plug and conidial suspension. Mycelial disc (5 mm) and a drop of 20 μl spore suspension (106 conidia/ml) were prepared from 7-day-old culture and placed on the fruit surface. The inoculated fruits were incubated at 27 ± 1°C for 10 days at 96.1% humidity. After 3 to 4 days of inoculation, brown to black spotted lesions were observed and coalesced to become black sunken lesions. Similar anthracnose symptoms were observed on all banana varieties tested. C. gloeosporioides was reisolated from the anthracnose lesions of all the inoculated fruit in which the cultural and morphological characteristics were the same as the original isolates. To our knowledge, this is the first report of C. gloeosporioides causing anthracnose of Musa spp. in Malaysia. References: (1) P. F. Cannon et al. Mycotaxon 104:189, 2008. (2) J. E. M. Mordue. Glomerella cingulata. CMI Description of Pathogenic Fungi and Bacteria, No. 315. CAB International,1971. (3) H. Prihastuti et al. Fungal Diversity 39:89, 2009.
    Matched MeSH terms: Agar
  20. Sakinah MAI, Latiffah Z
    Plant Dis, 2013 Aug;97(8):1110.
    PMID: 30722495 DOI: 10.1094/PDIS-09-12-0831-PDN
    Rambutan (Nephelium lappaceum L.) is among the tropical fruit grown in Malaysia and the demand for export rose in 2011. A fruit rot was observed between August and December 2011 from several areas in the states of Pulau Pinang and Perak, Malaysia. The symptoms initially appeared as light brown, water-soaked lesions that developed first in the pericarp and pulp, later enlarging and becoming dark brown. Greyish brown mycelia were observed on infected areas that turned yellowish at later stages of infection. Gliocephalotrichum bacillisporum was isolated from infected fruit by surface sterilization techniques. Conidia were mass-transferred onto potato dexstrose agar (PDA) plates and incubated at 27 ± 1°C. Tissue pieces (5 × 5 mm) excised from the margins between infected and healthy areas were then surface sterilized in 1% sodium hypochlorite for 3 to 5 min before being rinsed with distilled water, plated on PDA, and incubated at 27 ± 1°C for 7 days. Ten isolates of G. bacillisporum were obtained. Colonies on PDA were initially white before turning yellow with a feathery appearance. Microscopic characteristics on carnation leaf agar (CLA) consisted of hyaline conidia that were slightly ellipsoid to bacilliform with rounded apex ranging from 6.0 to 8.5 μm long and 2.0 to 2.5 μm wide. Conidiophores (70 to 130 μm long) were mostly single arising from large hypha approximately 13 to 16 μm. The conidiogenous structures were mostly quadriverticillate with dense, short, penicillate branches. The phialides were cylindrical and finger-like. Chlamydospores were present singly, in groups of 2 to 4, or in occasionally branched short chains and were brown in color with thick walls ranging from 11 to 13 μm. The cultural and morphological characteristics of G. bacillisporum isolates in the present study were very similar to previously published descriptions (1) except the conidiophores formed without sterile stipe extensions. All the G. bacillisporum isolates were deposited in culture collection at the Plant Pathology Lab, University Sains Malaysia, Penang. Molecular identification was accomplished from the ITS regions using ITS1 and ITS2 primers, and the β-tubulin gene using Bt2a and Bt2b primers (2). BLAST results from the ITS regions showed a 98 to 99% similarity with sequences of G. bacillisporum isolates reported in GenBank. Accession numbers of G. bacillisporum ITS regions: JX484850, JX484852, JX484853, JX484856, JX484858, JX484860, JX484862, JX484866, JX484867, and JX484868. The identity of G. bacillisporum isolates infecting rambutan was further confirmed by β-tubulin sequences (KC683909, KC683911, KC683912, KC683916, KC683919, KC683920, KC683923, KC683926, and KC683927), which showed 92 to 95% similarity with sequences of G. bacillisporum. Pathogenicity tests were also performed using mycelial plug (5 mm) and sprayed conidial suspensions (20 μl suspension of 106 conidia/ml) prepared from 7-day-old cultures. Inoculated fruits were incubated at 27 ± 1°C and after 10 days, similar rotting symptoms appeared on the fruit surface. The pathogen was reisolated from fruit rot lesions, thus fulfilling Koch's postulates, and tests were repeated twice. To our knowledge, this is the first report of G. bacillisporum causing fruit rot of rambutan (N. lappaceum L.) in Malaysia. References: (1) C. Decock et al. Mycologia 98:488, 2006. (2) N. L. Glass and G. C. Donaldson. Appl. Environ Microbiol. 61:1323, 1995.
    Matched MeSH terms: Agar
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