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  1. Al-Najjar BO, Saqallah FG, Abbas MA, Al-Hijazeen SZ, Sibai OA
    Eur J Med Chem, 2022 Jan 05;227:113924.
    PMID: 34731765 DOI: 10.1016/j.ejmech.2021.113924
    P2Y12 is a platelet surface protein which is responsible for the amplification of P2Y1 response. It plays a crucial role in platelet aggregation and thrombus formation through an ADP-induced platelet activation mechanism. Despite that P2Y12 platelets' receptor is an excellent target for developing antiplatelet agents, only five approved medications are currently in clinical use which are classified into thienopyridines and nucleoside-nucleotide derivatives. In the past years, many attempts for developing new candidates as P2Y12 inhibitors have been made. This review highlights the importance and the role of P2Y12 receptor as part of the coagulation cascade, its reported congenital defects, and the type of assays which are used to verify and measure its activity. Furthermore, an overview is given of the clinically approved medications, the potential naturally isolated inhibitors, and the synthesised candidates which were tested either in-vitro, in-vivo and/or clinically. Finally, we outline the in-silico attempts which were carried out using virtual screening, molecular docking and dynamics simulations in efforts of designing novel P2Y12 antagonists. Various phytochemical classes might be considered as a corner stone for the discovery of novel P2Y12 inhibitors, whereas a wide range of ring systems can be deliberated as leading scaffolds in that area synthetically and theoretically.
    Matched MeSH terms: Purinergic P2Y Receptor Antagonists/isolation & purification
  2. Fauzi FH, Hamzan NI, Rahman NA, Suraiya S, Mohamad S
    J Zhejiang Univ Sci B, 2021 4 13;21(12):961-976.
    PMID: 33843162 DOI: 10.1631/jzus.B2000161
    Worldwide there has been a significant increase in the incidence of oropharyngeal squamous cell carcinoma (OPSCC) etiologically attributed to oncogenic human papillomavirus (HPV). Reliable and accurate identification and detection tools are important as the incidence of HPV-related cancer is on the rise. Several HPV detection methods for OPSCC have been developed and each has its own advantages and disadvantages in regard to sensitivity, specificity, and technical difficulty. This review summarizes our current knowledge of molecular methods for detecting HPV in OPSCC, including HPV DNA/RNA polymerase chain reaction (PCR), loop-mediated isothermal amplification (LAMP), p16 immunohistochemistry (IHC), and DNA/RNA in situ hybridization (ISH) assays. This summary may facilitate the selection of a suitable method for detecting HPV infection, and therefore may help in the early diagnosis of HPV-related carcinoma to reduce its mortality, incidence, and morbidity.
    Matched MeSH terms: Alphapapillomavirus/isolation & purification*
  3. Altowayti WAH, Othman N, Al-Gheethi A, Dzahir NHBM, Asharuddin SM, Alshalif AF, et al.
    Molecules, 2021 Oct 13;26(20).
    PMID: 34684757 DOI: 10.3390/molecules26206176
    Sustainable wastewater treatment is one of the biggest issues of the 21st century. Metals such as Zn2+ have been released into the environment due to rapid industrial development. In this study, dried watermelon rind (D-WMR) is used as a low-cost adsorption material to assess natural adsorbents' ability to remove Zn2+ from synthetic wastewater. D-WMR was characterized using scanning electron microscope (SEM) and X-ray fluorescence (XRF). According to the results of the analysis, the D-WMR has two colours, white and black, and a significant concentration of mesoporous silica (83.70%). Moreover, after three hours of contact time in a synthetic solution with 400 mg/L Zn2+ concentration at pH 8 and 30 to 40 °C, the highest adsorption capacity of Zn2+ onto 1.5 g D-WMR adsorbent dose with 150 μm particle size was 25 mg/g. The experimental equilibrium data of Zn2+ onto D-WMR was utilized to compare nonlinear and linear isotherm and kinetics models for parameter determination. The best models for fitting equilibrium data were nonlinear Langmuir and pseudo-second models with lower error functions. Consequently, the potential use of D-WMR as a natural adsorbent for Zn2+ removal was highlighted, and error analysis indicated that nonlinear models best explain the adsorption data.
    Matched MeSH terms: Zinc/isolation & purification*
  4. Lim CH, Ng BH, Teh HL
    BMJ Case Rep, 2019 Nov 21;12(11).
    PMID: 31753829 DOI: 10.1136/bcr-2019-232828
    Matched MeSH terms: Calcium Pyrophosphate/isolation & purification*
  5. Shudirman S, Abang Kassim A, Shamsol Anuar NS, Utsumi M, Shimizu K, Muhammad Yuzir MA, et al.
    J Gen Appl Microbiol, 2021 Jul 31;67(3):92-99.
    PMID: 33642451 DOI: 10.2323/jgam.2020.08.001
    Musty odor production by actinomycetes is usually related to the presence of geosmin and 2-methylisoborneol (2-MIB), which are synthesized by enzymes encoded by the geoA and tpc genes, respectively. Streptomyces spp. strain S10, which was isolated from a water reservoir in Malaysia, has the ability to produce geosmin when cultivated in a basal salt (BS) solid medium, but no 2-MIB production occurred during growth in BS medium. Strain S10 could produce higher levels of geosmin when the phosphate concentration was limited to 0.05 mg/L, with a yield of 17.53 ± 3.12 ✕ 105 ng/L, compared with growth in BS medium. Interestingly, 2-MIB production was suddenly detected when the nitrate concentration was limited to 1.0 mg/L, with a yield of 1.4 ± 0.11 ✕ 105 ng/L. Therefore, it was concluded that phosphate- and nitrate-limiting conditions could induce the initial production of geosmin and 2-MIB by strain S10. Furthermore, a positive amplicon of geoA was detected in strain S10, but no tpc amplicon was detected by PCR analysis. Draft genome sequence analysis showed that one open reading frame (ORF) contained a conserved motif of geosmin synthase with 95% identity with geoA in Streptomyces coelicolor A3 (2). In the case of the tpc genes, it was found that one ORF showed 23% identity to the known tpc gene in S. coelicolor A3(2), but strain S10 lacked one motif in the N-terminus.
    Matched MeSH terms: Streptomyces/isolation & purification
  6. Abd Murad NB, Mohamed Nor NMI, Shohaimi S, Mohd Zainudin NAI
    J Appl Microbiol, 2017 Dec;123(6):1533-1546.
    PMID: 28891270 DOI: 10.1111/jam.13582
    AIMS: The aims of this study were to identify the Fusarium isolates based on translation elongation factor (tef) 1α sequence, to determine the genetic diversity among isolates and species using selected microsatellite markers and to examine the pathogenicity of Fusarium isolates causing fruit rot disease of banana.

    METHODS AND RESULTS: One-hundred and thirteen microfungi isolates were obtained from fruit rot infected banana in Peninsular Malaysia. However, this study was focused on the dominant number of the discovered microfungi that belongs to the genus Fusarium; 48 isolates of the microfungi have been identified belonging to 11 species of Fusarium, namely Fusarium incarnatum, Fusarium equiseti, Fusarium camptoceras, Fusarium solani, Fusarium concolor, Fusarium oxysporum, Fusarium proliferatum, Fusarium verticillioides, Fusarium sacchari, Fusarium concentricum and Fusarium fujikuroi. All Fusarium isolates were grouped into their respective clades indicating their similarities and differences in genetic diversity among isolates. Out of 48 Fusarium isolates tested, 42 isolates caused the fruit rot symptom at different levels of severity based on Disease Severity Index (DSI). The most virulent isolate was F. proliferatum B2433B with DSI of 100%.

    CONCLUSIONS: All the isolated Fusarium species were successfully identified and some of them were confirmed as the causal agents of pre- and postharvest fruit rot in banana across Peninsular Malaysia.

    SIGNIFICANCE AND IMPACT OF THE STUDY: Our results will provide additional information regarding new report of Fusarium species in causing banana fruit rot and in the search of potential biocontrol agent of the disease.

    Matched MeSH terms: Fusarium/isolation & purification
  7. Péré-Védrenne C, Flahou B, Loke MF, Ménard A, Vadivelu J
    Helicobacter, 2017 Sep;22 Suppl 1.
    PMID: 28891140 DOI: 10.1111/hel.12407
    The current article is a review of the most important and relevant literature published in 2016 and early 2017 on non-Helicobacter pylori Helicobacter infections in humans and animals, as well as interactions between H. pylori and the microbiota of the stomach and other organs. Some putative new Helicobacter species were identified in sea otters, wild boars, dogs, and mice. Many cases of Helicobacter fennelliae and Helicobacter cinaedi infection have been reported in humans, mostly in immunocompromised patients. Mouse models have been used frequently as a model to investigate human Helicobacter infection, although some studies have investigated the pathogenesis of Helicobacters in their natural host, as was the case for Helicobacter suis infection in pigs. Our understanding of both the gastric and gut microbiome has made progress and, in addition, interactions between H. pylori and the microbiome were demonstrated to go beyond the stomach. Some new approaches of preventing Helicobacter infection or its related pathologies were investigated and, in this respect, the probiotic properties of Saccharomyces, Lactobacillus and Bifidobacterium spp. were confirmed.
    Matched MeSH terms: Helicobacter/isolation & purification*
  8. Chong KW, Yeap JS, Lim SH, Weber JF, Low YY, Kam TS
    J Nat Prod, 2017 11 22;80(11):3014-3024.
    PMID: 29087707 DOI: 10.1021/acs.jnatprod.7b00621
    Reexamination of the absolute configuration of recently isolated eburnane alkaloids from Malaysian Kopsia and Leuconotis species by X-ray diffraction analysis and ECD/TDDFT has revealed the existence of biosynthetic enantiodivergence. Three different scenarios are discerned with respect to the composition of the enantiomeric eburnane alkaloids in these plants: first, where the new eburnane congeners possess the same C-20, C-21 absolute configurations as the common eburnane alkaloids (eburnamonine, eburnamine, isoeburnamine, eburnamenine) occurring in the same plant; second, where the new eburnane congeners possess opposite or enantiomeric C-20, C-21 absolute configurations compared to the common eburnane alkaloids found in the same plant; and, third, where the four common eburnane alkaloids were isolated as racemic or scalemic mixtures, while the new eburnane congeners were isolated as pure enantiomers with a common C-20, C-21 configuration (20α, 21α). Additionally, the same Kopsia species (K. pauciflora) found in two different geographical locations (Peninsular Malaysia and Malaysian Borneo) showed different patterns in the composition of the enantiomeric eburnane alkaloids. Revision of the absolute configurations of a number of new eburnane congeners (previously assigned based on the assumption of a common biogenetic origin to that of the known eburnane alkaloids co-occurring in the same plant) is required based on the present results.
    Matched MeSH terms: Indole Alkaloids/isolation & purification*
  9. Danderson CA, Downie SR, Hermann M
    Mol Phylogenet Evol, 2018 01;118:286-305.
    PMID: 29017853 DOI: 10.1016/j.ympev.2017.10.006
    The Arracacia clade (Apiaceae, Apioideae) is a heterogeneous assemblage of 12 genera, comprising 111 known species distributed in high montane temperate and sub-alpine habitats of meso- and South America. Previous studies have indicated that the genera Arracacia, Coulterophytum, and Prionosciadium are polyphyletic, but for the most part relationships among the members of the clade are largely unknown. Initially, cladistic analyses of nrDNA ITS sequences were carried out on 212 accessions (122 taxa), representing 92 species of the Arracacia clade and outgroups from the closely-related páramo genera Cotopaxia, Niphogeton, and Perissocoeleum and members of the Perennial Endemic North American clade and its allies. Using the ITS results to inform sampling of a small subset of taxa, a pilot study examining the phylogenetic utility of 20 noncoding chloroplast loci was subsequently performed to identify those regions most useful at resolving relationships. A cost-benefit analysis determined that five loci (trnQ-5'rps16, trnD-trnT, rpl32-trnL, psbD-trnT, ndhA intron) would maximize resolution and branch support in the clade. Cladistic analyses of four of these loci (trnQ-5'rps16, trnD-trnT, rpl32-trnL, ndhA intron) and the ITS region, separately and combined, revealed that Arracacia, Coaxana, Coulterophytum, Prionosciadium, and Rhodosciadium are each polyphyletic and that Donnellsmithia and Myrrhidendron are each monophyletic. Although most relationships in the Arracacia clade and among the closely-related genera Cotopaxia, Niphogeton, and Perissocoeleum are poorly resolved and supported, ten groups are recognized for future revisionary studies. Polyploidy and rapid species radiation have likely confounded generic circumscriptions and interpretation of relationships.
    Matched MeSH terms: DNA, Chloroplast/isolation & purification
  10. Karimi E, Ghorbani Nohooji M, Habibi M, Ebrahimi M, Mehrafarin A, Khalighi-Sigaroodi F
    Nat Prod Res, 2018 Aug;32(16):1991-1995.
    PMID: 28774179 DOI: 10.1080/14786419.2017.1359171
    The antioxidant activities of crude extract fractions using Hexane, Chloroform, Ethyl Acetate, Butanol and Water of Clematis orientalis and Clematis ispahanica were investigated. 1,1-diphenyl-2-picryl-hydrazyl (DPPH) assay and the ferric reducing/antioxidant potential (FRAP) were used to evaluate the antioxidant capacity. The total phenolics were found to be 4.37-9.38 and 1.32-11.37 mg gallic acid equivalents (GAE)/g in different fractions for C. orientalis and C. ispahanica, respectively. The ethyl acetate fraction of C. orientalis and chloroform fraction of C. ispahanica showed the highest DPPH and FRAP activities at a concentration of 300 μg/mL. The predominant phenolic compounds identified by HPLC in C. orientalis were Resorcinol (603.5 μg/g DW) in chloroform fraction and Ellagic acid (811.7 μg/g DW) in chloroform fraction of C. ispahanica.
    Matched MeSH terms: Antioxidants/isolation & purification*
  11. Lee HX, Ahmad F, Saad B, Ismail MN
    Prep Biochem Biotechnol, 2017 Nov 26;47(10):998-1007.
    PMID: 28857669 DOI: 10.1080/10826068.2017.1365250
    Date fruits are well known to be very nutritious. Nevertheless, the protein contents of the fruit, particularly the seed and flesh, are still understudied, largely due to their difficult physical characteristics. This study was conducted to compare three different protein extraction methods which were the trichloroacetic acid (TCA)-acetone (TCA-A), phenol (Phe), and TCA-acetone-phenol (TCA-A-Phe), and to perform proteomic analysis on date palm seed and flesh. Phe extraction method showed the highest protein yields for both seed (8.26 mg/g) and flesh (1.57 mg/g). Through sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Phe, and TCA-A-Phe extraction methods were shown to be efficient in removing interfering compounds and gave well-resolved bands over a wide range of molecular weights. Following liquid chromatography-tandem mass spectrometry analysis, about 50-64% of extracted proteins were identified with known functions including those involved in glycolysis, Krebs cycle, defense, and storage. Phe protein extraction method was proven to be the optimal method for date flesh and seed.
    Matched MeSH terms: Plant Proteins/isolation & purification
  12. Naderali N, Nejat N, Vadamalai G, Davis RE, Wei W, Harrison NA, et al.
    Int J Syst Evol Microbiol, 2017 Oct;67(10):3765-3772.
    PMID: 28905707 DOI: 10.1099/ijsem.0.002187
    Landscape-grown foxtail palm (Wodyetia bifurcata A. K. Irvine) trees displaying symptoms of severe foliar chlorosis, stunting, general decline and mortality reminiscent of coconut yellow decline disease were observed in Bangi, Malaysia, during 2012. DNA samples from foliage tissues of 15 symptomatic palms were analysed by employing a nested PCR assay primed by phytoplasma universal ribosomal RNA operon primer pairs, P1/P7 followed by R16F2n/R2. The assay yielded amplicons of a single band of 1.25 kb from DNA samples of 11 symptomatic palms. Results from cloning and sequence analysis of the PCR-amplified 16S rRNA gene segments revealed that, in three palms, three mutually distinct phytoplasmas comprising strains related to 'Candidatus Phytoplasma asteris' and 'Candidatus Phytoplasma cynodontis', as well as a novel phytoplasma, were present as triple infections. The 16S rRNA gene sequence derived from the novel phytoplasma shared less than 96 % nucleotide sequence identity with that of each previously describedspecies of the provisional genus 'Ca. Phytoplasma', justifying its recognition as the reference strain of a new taxon, 'Candidatus Phytoplasma wodyetiae'. Virtual RFLP profiles of the R16F2n/R2 portion of the 16S rRNA gene and the pattern similarity coefficient value (0.74) supported the delineation of 'Ca. Phytoplasma wodyetiae' as the sole representative subgroup A member of a new phytoplasma ribosomal group, 16SrXXXVI.
    Matched MeSH terms: Phytoplasma/isolation & purification
  13. Ser HL, Tan WS, Ab Mutalib NS, Yin WF, Chan KG, Goh BH, et al.
    Braz J Microbiol, 2017 09 06;49(1):13-15.
    PMID: 28927873 DOI: 10.1016/j.bjm.2017.01.013
    As the largest genus in Actinobacteria family, Streptomyces species have the ability to synthesize numerous compounds of diverse structures with bioactivities. Streptomyces mangrovisoli MUSC 149T was previously isolated as a novel streptomycete from mangrove forest in east coast of Peninsular Malaysia. The high quality draft genome of MUSC 149T comprises 9,165,825bp with G+C content of 72.5%. Through bioinformatics analysis, 21 gene clusters identified in the genome were associated with the production of bioactive secondary metabolites. The presence of these biosynthetic gene clusters in MUSC 149T suggests the potential exploitation of the strain for production of medically important compounds.
    Matched MeSH terms: Streptomyces/isolation & purification*
  14. Auta HS, Emenike CU, Fauziah SH
    Environ Pollut, 2017 Dec;231(Pt 2):1552-1559.
    PMID: 28964604 DOI: 10.1016/j.envpol.2017.09.043
    The continuous accumulation of microplastics in the environment poses ecological threats and has been an increasing problem worldwide. In this study, eight bacterial strains were isolated from mangrove sediment in Peninsular Malaysia to mitigate the environmental impact of microplastics and develop a clean-up option. The bacterial isolates were screened for their potential to degrade UV-treated microplastics from polyethylene (PE), polyethylene terephthalate (PET), polypropylene (PP), and polystyrene (PS). Only two isolates, namely, Bacillus cereus and Bacillus gottheilii, grew on a synthetic medium containing different microplastic polymers as the sole carbon source. A shake flask experiment was carried out to further evaluate the biodegradability potential of the isolates. Degradation was monitored by recording the weight loss of microplastics and the growth pattern of the isolates in the mineral medium. The biodegradation extent was validated by assessment of the morphological and structural changes through scanning electron microscopy and Fourier transform infrared spectroscopy analyses. The calculated weight loss percentages of the microplastic particles by B. cereus after 40 days were 1.6%, 6.6%, and 7.4% for PE, PET, and PS, respectively. B. gottheilii recorded weight loss percentages of 6.2%, 3.0%, 3.6%, and 5.8% for PE, PET, PP, and PS, respectively. The designated isolates degraded the microplastic material and exhibited potential for remediation of microplastic-contaminated environment. Biodegradation tests must be conducted to characterize the varied responses of microbes toward pollutants, such as microplastics. Hence, a novel approach for biodegradation of microplastics must be developed to help mitigate the environmental impact of plastics and microplastic polymers.
    Matched MeSH terms: Bacillus/isolation & purification*
  15. Thevarajoo S, Selvaratnam C, Chan KG, Goh KM, Chong CS
    Braz J Microbiol, 2017 07 19;49(1):10-12.
    PMID: 28778371 DOI: 10.1016/j.bjm.2017.03.013
    Vitellibacter aquimaris D-24T (=KCTC 42708T=DSM 101732T), a halophilic marine bacterium, was isolated from seawater collected from Desaru beach, Malaysia. Here, we present the draft genome sequence of D-24T with a genome size of approximately 3.1Mbp and G+C content of 39.93%. The genome of D-24T contains genes involved in reducing a potent greenhouse gas (N2O) in the environment and the degradation of proteinaceous compounds. Genome availability will provide insights into potential biotechnological and environmental applications of this bacterium.
    Matched MeSH terms: Flavobacteriaceae/isolation & purification
  16. Seow SL, Naidu M, Sabaratnam V, Vidyadaran S, Wong KH
    Int J Med Mushrooms, 2017;19(5):405-418.
    PMID: 28845770 DOI: 10.1615/IntJMedMushrooms.v19.i5.30
    In Malaysia and China, the sclerotium of Lignosus rhinocerotis is used by local communities and traditional medicine practitioners as a general tonic and remedy to treat a variety of ailments, including inflammation-associated disorders. In this study, 10 samples from different preparations of L. rhinocerotis sclerotium, including a hot aqueous extract (HAE), an ethanol extract (EE), fractions from the HAE and EE, and crude polysaccharides, were tested for their in vitro cytotoxic and nitric oxide (NO) inhibitory activities in lipopolysaccharide (LPS)--stimulated BV2 microglia. Of the 10 samples tested, HAE was the least cytotoxic toward BV2 microglia, with a half-maximal inhibitory concentration of 176.23 ± 2.64 mg/mL at 24 hours of incubation and 20.01 ± 1.69 mg/ mL at 48 hours of incubation. The inhibition of NO production was explored by pretreatment of BV2 microglia with samples at 2 incubation time points (4 and 24 hours) before the stimulation by LPS for 24 hours. After 24 hours of pretreatment, 8 of the 10 samples inhibited NO production by 50% or more, and cytotoxic effects were not observed. Among the 8 active samples, 500 µg/mL of HAE, 250 µg/mL of an n-butanol fraction of the HAE, and 250 µg/mL of an ethyl acetate fraction of HAE showed maximum inhibition of NO production by 88.95%, 86.50%, and 85.93%, respectively. These results suggest that the L. rhinocerotis sclerotium may contain secondary metabolites that have the potential to inhibit NO production.
    Matched MeSH terms: Cell Extracts/isolation & purification
  17. Khoo JJ, Lim FS, Tan KK, Chen FS, Phoon WH, Khor CS, et al.
    J Med Entomol, 2017 09 01;54(5):1444-1448.
    PMID: 28874019 DOI: 10.1093/jme/tjx131
    Spirochetes from the Borrelia genus are known to cause diseases in humans, namely Lyme disease and relapsing fever. These organisms are commonly transmitted to humans by arthropod vectors including ticks, mite, and lice. Here, we report the molecular detection of a Borrelia sp. from a Haemaphysalis hystricis Supino tick collected from wildlife in an Orang Asli settlement in Selangor, Malaysia. Phylogenetic analyses of partial 16s rRNA and flaB gene sequences revealed that the Borrelia sp. is closely related to the relapsing fever group borreliae, Borrelia lonestari, Borrelia miyamotoi, and Borrelia theileri, as well as a number of uncharacterized Borrelia sp. from ticks in Portugal and Japan. To our knowledge, this is the first report of a Borrelia sp. detected in H. hystricis, and in Malaysia. The zoonotic potential of this Borrelia sp. merits further investigation.
    Matched MeSH terms: Borrelia/isolation & purification*
  18. Mohan Jacob D, Lee CY, Arshad SS, Selvarajah GT, Bande F, Ong BL, et al.
    Trop Anim Health Prod, 2018 Apr;50(4):733-739.
    PMID: 29243138 DOI: 10.1007/s11250-017-1489-z
    Several strains of porcine bocaviruses have been reported worldwide since their first detection in Sweden in 2009. Subsequently, the virus has been reported to be associated with gastrointestinal and respiratory signs in weaner and grower pigs. Although Malaysia is host to a self-sufficient swine livestock industry, there is no study that describes porcine bocavirus in the country. This report is the first to describe porcine bocavirus (PBoV) in Malaysian swine herds. PBoV was identified in various tissues from sick and runt pigs using the conventional PCR method with primers targeting conserved regions encoding for the nonstructural protein (NS1) gene. Out of 103 samples tested from 17 pigs, 32 samples from 15 pigs were positive for porcine bocavirus. In addition, a higher detection rate was identified from mesenteric lymph nodes (52.9%), followed by tonsil (37.0%), and lungs (33.3%). Pairwise comparison and phylogenetic analyses based on a 658-bp fragment of NS1 gene revealed that the Malaysian PBoV strains are highly similar to PBoV3 isolated in Minnesota, USA. The presence of porcine bocavirus in Malaysia and their phylogenetic bond was marked for the first time by this study. Further studies will establish the molecular epidemiology of PBoV in Malaysia and clarify pathogenicity of the local isolates.
    Matched MeSH terms: Bocavirus/isolation & purification*
  19. Wassermann M, Raisch L, Lyons JA, Natusch DJD, Richter S, Wirth M, et al.
    PLoS One, 2017;12(11):e0187984.
    PMID: 29131856 DOI: 10.1371/journal.pone.0187984
    We examined Sarcocystis spp. in giant snakes from the Indo-Australian Archipelago and Australia using a combination of morphological (size of sporocyst) and molecular analyses. We amplified by PCR nuclear 18S rDNA from single sporocysts in order to detect mixed infections and unequivocally assign the retrieved sequences to the corresponding parasite stage. Sarcocystis infection was generally high across the study area, with 78 (68%) of 115 examined pythons being infected by one or more Sarcocystis spp. Among 18 randomly chosen, sporocyst-positive samples (11 from Southeast Asia, 7 from Northern Australia) the only Sarcocystis species detected in Southeast Asian snakes was S. singaporensis (in reticulated pythons), which was absent from all Australian samples. We distinguished three different Sarcocystis spp. in the Australian sample set; two were excreted by scrub pythons and one by the spotted python. The sequence of the latter is an undescribed species phylogenetically related to S. lacertae. Of the two Sarcocystis species found in scrub pythons, one showed an 18S rRNA gene sequence similar to S. zamani, which is described from Australia for the first time. The second sequence was identical/similar to that of S. nesbitti, a known human pathogen that was held responsible for outbreaks of disease among tourists in Malaysia. The potential presence of S. nesbitti in Australia challenges the current hypothesis of a snake-primate life cycle, and would have implications for human health in the region. Further molecular and biological characterizations are required to confirm species identity and determine whether or not the Australian isolate has the same zoonotic potential as its Malaysian counterpart. Finally, the absence of S. nesbitti in samples from reticulated pythons (which were reported to be definitive hosts), coupled with our phylogenetic analyses, suggest that alternative snake hosts may be responsible for transmitting this parasite in Malaysia.
    Matched MeSH terms: Sarcocystis/isolation & purification*
  20. Wong YP, Othman S, Lau YL, Radu S, Chee HY
    J Appl Microbiol, 2018 Mar;124(3):626-643.
    PMID: 29165905 DOI: 10.1111/jam.13647
    Loop-mediated isothermal amplification (LAMP) amplifies DNA with high specificity, efficiency and rapidity under isothermal conditions by using a DNA polymerase with high displacement strand activity and a set of specifically designed primers to amplify targeted DNA strands. Following its first discovery by Notomi et al. ( Nucleic Acids Res 28: E63), LAMP was further developed over the years which involved the combination of this technique with other molecular approaches, such as reverse transcription and multiplex amplification for the detection of infectious diseases caused by micro-organisms in humans, livestock and plants. In this review, available types of LAMP techniques will be discussed together with their applications in detection of various micro-organisms. Up to date, there are varieties of LAMP detection methods available including colorimetric and fluorescent detection, real-time monitoring using turbidity metre and detection using lateral flow device which will also be highlighted in this review. Apart from that, commercialization of LAMP technique had also been reported such as lyophilized form of LAMP reagents kit and LAMP primer sets for detection of pathogenic micro-organisms. On top of that, advantages and limitations of this molecular detection method are also described together with its future potential as a diagnostic method for infectious disease.
    Matched MeSH terms: DNA/isolation & purification
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