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Simultaneous detection of 12 mycotoxins in cereals using RP-HPLC-PDA-FLD with PHRED and a post-column derivatization system

Soleimany F, Jinap S, Rahmani A, Khatib A

Food Addit Contam Part A Chem Anal Control Expo Risk Assess, 2011 Apr;28(4):494-501.
PMID: 21337232 DOI: 10.1080/19440049.2010.551547

A new method for the simultaneous quantification of 12 mycotoxins was developed and optimized using reverse phase high performance liquid chromatography (RP-HPLC) with a photodiode array (PDA) and fluorescence detector (FLD), a photochemical reactor for enhanced detection (PHRED) and post-column derivatization. The mycotoxins included aflatoxins (AFB(1), AFB(2), AFG(1), and AFG(2)), ochratoxin A (OTA), zearalenone (ZEA), deoxynivalenol (DON), fumonisins (FB(1), FB(2), and FB(3)), T-2 and HT-2 toxins. A double sample extraction with a phosphate-buffered saline solution (PBS) and methanol was used for co-extraction of mycotoxins, and a multifunctional immunoaffinity column was used for cleanup. Optimum conditions for separation of the mycotoxins were obtained to separate 12 mycotoxins in FLD and PDA chromatograms with a high resolution. The method gave recoveries in the range 72-111% when applied to spiked corn samples. The limits of detection (LOD) were 0.025 ng/g for AFB(1) and AFG(1), 0.012 ng/g for AFB(2) and AFG(2), 0.2 ng/g for OTA, 1.5 ng/g for ZEA, 6.2 ng/g for FB(1), FB(3) and HT-2 toxin, 9.4 ng/g for FB(2) and T-2 toxin, and 18.7 ng/g for DON. In addition, the limits of quantification (LOQ) ranged from 0.04 ng/g for AFB(2) and AFG(2) to 62 ng/g for DON. The method was successfully applied to the determination of these mycotoxins in 45 cereal samples obtained from the Malaysian market. The results indicated that the method can be applied for the multi-mycotoxin determination of cereals.

Matched MeSH terms: Mycotoxins/isolation & purification
Detection of immunogenic parasite and host-specific proteins in the sera of active and chronic individuals infected with Toxoplasma gondii

Yeng C, Osman E, Mohamed Z, Noordin R

Electrophoresis, 2010 Dec;31(23-24):3843-9.
PMID: 21080484 DOI: 10.1002/elps.201000038

Toxoplasma gondii infection in pregnant women may result in abortion and foetal abnormalities, and may be life-threatening in immunocompromised hosts. To identify the potential infection markers of this disease, 2-DE and Western blot methods were employed to study the parasite circulating antigens and host-specific proteins in the sera of T. gondii-infected individuals. The comparisons were made between serum protein profiles of infected (n=31) and normal (n=10) subjects. Antigenic proteins were identified by immunoblotting using pooled sera and monoclonal anti-human IgM-HRP. Selected protein spots were characterised using mass spectrometry. Prominent differences were observed when serum samples of T. gondii-infected individuals and normal controls were compared. A significant up-regulation of host-specific proteins, α(2)-HS glycoprotein and α(1)-B glycoprotein, was also observed in the silver-stained gels of both active and chronic infections. However, only α(2)-HS glycoprotein and α(1)-B glycoprotein in the active infection showed immunoreactivity in Western blots. In addition, three spots of T. gondii proteins were detected, namely (i) hypothetical protein chrXII: 3984434-3 TGME 49, (ii) dual specificity protein phosphatase, catalytic domain TGME 49 and (iii) NADPH-cytochrome p450 reductase TGME 49. Thus, 2-DE approach followed by Western blotting has enabled the identification of five potential infection markers for the diagnosis of toxoplasmosis: three are parasite-specific proteins and two are host-specific proteins.

Matched MeSH terms: Toxoplasma/isolation & purification*
Leachate characterization in semi-aerobic and anaerobic sanitary landfills: a comparative study

Aziz SQ, Aziz HA, Yusoff MS, Bashir MJ, Umar M

J Environ Manage, 2010 Dec;91(12):2608-14.
PMID: 20739117 DOI: 10.1016/j.jenvman.2010.07.042

This study analyzes and compares the results of leachate composition at the semi-aerobic Pulau Burung Landfill Site (PBLS) (unaerated pond and intermittently aerated pond) and the anaerobic Kulim Sanitary Landfill in the northern region of Malaysia. The raw samples were collected and analyzed for twenty parameters. The average values of the parameters such as phenols (1.2, 6.7, and 2.6 mg/L), total nitrogen (448, 1200, and 300 mg/L N-TN), ammonia-N (542, 1568, and 538 mg/L NH(3)-N), nitrite (91, 49, and 52 mg/L NO(2)(-)-N), total phosphorus (21, 17, and 19 mg/L), BOD(5) (83, 243, and 326 mg/L), COD (935, 2345, and 1892 mg/L), BOD(5)/COD (0.096,0.1124,0.205%), pH (8.20, 8.28, and 7.76), turbidity (1546, 180, and 1936 Formazin attenuation units (FAU)), and color (3334, 3347, and 4041 Pt Co) for leachate at the semi-aerobic PBLS (unaerated and intermittently aerated) and the anaerobic Kulim Sanitary Landfill were recorded, respectively. The obtained results were compared with previously published data and data from the Malaysia Environmental Quality Act 1974. The results indicated that Pulau Burung leachate was more stabilized compared with Kulim leachate. Furthermore, the aeration process in PBLS has a considerable effect on reducing the concentration of several pollutants. The studied leachate requires treatment to minimize the pollutants to an acceptable level prior to discharge into water courses.

Matched MeSH terms: Escherichia coli/isolation & purification
Resveratrol oligomers from Vatica albiramis

Abe N, Ito T, Ohguchi K, Nasu M, Masuda Y, Oyama M, et al.

J Nat Prod, 2010 Sep 24;73(9):1499-506.
PMID: 20735051 DOI: 10.1021/np1002675

Five new stilbenoids, vatalbinosides A-E (1-5), and 13 known compounds (6-18) were isolated from the stem of Vatica albiramis. The effects of these new compounds on interleukin-1β-induced production of matrix metalloproteinase-1 (MMP-1) in human dermal fibroblasts were examined. Three resveratrol tetramers, (-)-hopeaphenol (6), vaticanol C (13), and stenophyllol C (14), were identified as strong inhibitors of MMP-1 production.

Matched MeSH terms: Stilbenes/isolation & purification*
Use of Chlorella vulgaris for bioremediation of textile wastewater

Lim SL, Chu WL, Phang SM

Bioresour Technol, 2010 Oct;101(19):7314-22.
PMID: 20547057 DOI: 10.1016/j.biortech.2010.04.092

The potential application of Chlorella vulgaris UMACC 001 for bioremediation of textile wastewater (TW) was investigated using four batches of cultures in high rate algae ponds (HRAP) containing textile dye (Supranol Red 3BW) or TW. The biomass attained ranged from 0.17 to 2.26 mg chlorophyll a/L while colour removal ranged from 41.8% to 50.0%. There was also reduction of NH(4)-N (44.4-45.1%), PO(4)-P (33.1-33.3%) and COD (38.3-62.3%) in the TW. Supplementation of the TW with nutrients of Bold's Basal Medium (BBM) increased biomass production but did not improve colour removal or reduction of pollutants. The mechanism of colour removal by C. vulgaris is biosorption, in accordance with both the Langmuir and Freundlich models. The HRAP using C. vulgaris offers a good system for the polishing of TW before final discharge.

Matched MeSH terms: Water Pollutants, Chemical/isolation & purification
Developing a validated liquid chromatography-mass spectrometric method for the simultaneous analysis of five bioactive quassinoid markers for the standardization of manufactured batches of Eurycoma longifolia Jack extract as antimalarial medicaments

Teh CH, Murugaiyah V, Chan KL

J Chromatogr A, 2011 Apr 8;1218(14):1861-77.
PMID: 21367427 DOI: 10.1016/j.chroma.2011.02.014

An extensive comparative study on the electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI) mass spectrometry using automated flow injection analysis (FIA), was performed on eurycomanone (1), 13α(21)-epoxyeurycomanone (2), eurycomanol (3), eurycomanol-2-O-β-d-glucopyranoside (4), and 13,21-dihydroeurycomanone (5), the bioactive markers isolated from Eurycoma longifolia. The effects of eluent mixture (methanol or acetonitrile in water) and acidic modifiers (acetic acid, formic acid and trifluoroacetic acid) on the ionization efficiency of the markers were also investigated. The ESI in the positive ion mode with methanol containing 0.1% (v/v) acetic acid was selected for the subsequent optimization of nebulizer pressure, dry gas flow, dry gas temperature and capillary voltage to improve the sensitivity of the total ion chromatogram (TIC). Fragmentation of the analytes was further investigated by varying the capillary exit offset voltage and fragmentation amplitude in positive mode of ESI. The detection limits (LODs) were determined in isolation mode (selected ion monitoring, SIM). Their limits of detection (LODs) ranged between 0.03 and 0.1μgmL(-1) while the intra-day and inter-day precisions were less than 5.72% and 4.82%, respectively. The method was next applied for the simultaneous analysis of the markers to standardize various batches of manufactured extracts of E. longifolia for potential use as antimalarial products. Multiple Reaction Monitoring (MRM) mode was used for the quantification of analytes which gave protonated molecular ion, [M+H](+). For those without pseudo-molecular ions, SIM mode was used to quantify the analytes. The batches contained 5.65-9.95% of eurycomanone (1), 5.21-19.75% of eurycomanol (3) and 7.59-19.95% of eurycomanol-2-O-β-d-glucopyranoside (4) as major quassinoids whereas, 13α(21)-epoxyeurycomanone (2), and 13,21-dihydroeurycomanone (5) were much lower in concentrations of 0.78-3.90% and 0.47-1.76%, respectively.

Matched MeSH terms: Quassins/isolation & purification
Strychnan and secoangustilobine A type alkaloids from Alstonia spatulata. Revision of the C-20 configuration of scholaricine

Tan SJ, Low YY, Choo YM, Abdullah Z, Etoh T, Hayashi M, et al.

J Nat Prod, 2010 Nov 29;73(11):1891-7.
PMID: 21043460 DOI: 10.1021/np100552b

A total of 25 alkaloids were isolated from the leaf and stem-bark extracts of Alstonia spatulata, of which five are new alkaloids of the strychnan type (alstolucines A-E, 1-5) and the other, a new alkaloid of the secoangustilobine A type (alstolobine A, 6). The structures of these alkaloids were established using NMR and MS analysis and, in the case of alstolucine B (2), also confirmed by X-ray diffraction analysis. A reinvestigation of the stereochemical assignment of scholaricine (13) by NMR and X-ray analyses indicated that the configuration at C-20 required revision. Alkaloids 1, 2, 6, 7, 9, 10, and 13 reversed multidrug resistance in vincristine-resistant KB cells.

Matched MeSH terms: Indole Alkaloids/isolation & purification*
The manual MGIT system for the detection of M. tuberculosis in respiratory specimens: an experience in the University Malaya Medical Centre

Fadzilah MN, Ng KP, Ngeow YF

Malays J Pathol, 2009 Dec;31(2):93-7.
PMID: 20514851

A prospective study was conducted on 510 respiratory specimens for the presence of M. tuberculosis detected by direct acid-fast bacilli (AFB) smear examination, culture in the Manual Mycobacteria Growth Indicator Tube (BBL MGIT, Becton-Dickinson) and culture on Lowenstein-Jensen (LJ) medium. From positive BBL MGIT tubes, Ziehl-Neelsen and Gram stains were performed and subcultures were put up on LJ medium. A total of 101 (19.8%) specimens were positive by the BBL MGIT, 60 (11.8%) by primary LJ medium culture, 31 (6.1%) by direct smear examination and 29 (5.7%) by all three methods. Using primary LJ culture as the gold standard, the sensitivity and specificity of the BBL MGIT were 90% and 89.6% respectively but the sensitivity of AFB smear microscopy was only 48.3%. About half (51.1%) of the BBL MGIT false positives were due to contamination by non-AFB bacteria. The remaining false positives comprised specimens that were AFB microscopy positive but LJ culture negative. Of the AFB isolates obtained on LJ primary and sub-cultures, almost all (93.3%) were identified as Mycobacterium tuberculosis complex. The mean time-to-detection was significantly shorter (p < 0.0001) for the BBL MGIT than for LJ culture. For the former, positive results were available within 14 days for both AFB smear-positive and AFB smear-negative specimens. On the average, positive results were obtained 1.8 days earlier for direct AFB smear-positive samples than for AFB smear-negative samples. On the other hand, positive growth on LJ medium appeared after at least 33 days of incubation. These findings suggest that the BBL MGIT system will be a suitable alternative to LJ culture for the routine diagnosis of pulmonary tuberculosis, but a combination of liquid and solid cultures is still required for the highest diagnostic accuracy.

Matched MeSH terms: Mycobacterium tuberculosis/isolation & purification*
Effects of mitragynine from Mitragyna speciosa Korth leaves on working memory

Apryani E, Hidayat MT, Moklas MA, Fakurazi S, Idayu NF

J Ethnopharmacol, 2010 Jun 16;129(3):357-60.
PMID: 20371280 DOI: 10.1016/j.jep.2010.03.036

AIM OF THE STUDY: Mitragyna speciosa Korth from Rubiaceae family is a tropical plant indigenous to Southeast Asia particularly in Thailand, Peninsular of Malaysia and Indonesia. The leaves have been used by natives for their opium-like effect and cocaine-like stimulant ability to combat fatigue and enhance tolerance to hard work. However there is no scientific information about the effect of mitragynine on the cognitive performances. This study is designed to examine the working memory effects of mitragynine which is extracted from Mitragyna speciosa mature leaves.
MATERIALS AND METHODS: The cognitive effect was studied using object location task and the motor activity in open-field test. Mitragynine 5, 10 and 15 mg/kg and were administered by intraperitoneal (IP) for 28 consecutive days and evaluated on day 28 after the last dose treatment. Scopolamine was used as the control positive drug.
RESULTS: In this study there is prominent effects on horizontal locomotor activity was observed. Mitragynine significantly reduced locomotor activity in open-field test compared with vehicle. In object location task mitragynine (5, 10 and 15 mg/kg) did not showed any significances discrimination between the object that had changed position than the object that had remain in a constant position.
CONCLUSION: Our results suggest that chronic administration of mitragynine can altered the cognitive behavioral function in mice.

Matched MeSH terms: Secologanin Tryptamine Alkaloids/isolation & purification
Fulltext Dissemination of streptococcal pyrogenic exotoxin G (spegg) with an IS-like element in fish isolates of Streptococcus dysgalactiae

Abdelsalam M, Chen SC, Yoshida T

FEMS Microbiol Lett, 2010 Aug 1;309(1):105-13.
PMID: 20528946 DOI: 10.1111/j.1574-6968.2010.02024.x

The Lancefield group C alpha-hemolytic Streptococcus dysgalactiae ssp. dysgalactiae (GCSD) causes systemic granulomatous inflammatory disease and high mortality rates in infected fish. Superantigen and streptolysin S genes are the most important virulence factors contributing to an invasive streptococcal infection. PCR amplification revealed that all strains isolated from moribund fish harbored the streptolysin S structural gene (sagA). GCSD fish isolates were PCR negative for emm, speA, speB, speC, speM, smeZ, and ssa. However, the size of the streptococcal pyrogenic exotoxin G (spegg) locus, a superantigen, in positive S. dysgalactiae fish and pig strains was variable. The ORF of the spegg locus of 26 GCSD fish strains and one GCSD pig strain was inserted with IS981SC. Interestingly, the ORF of the spegg locus of two fish strains of GCSD collected in Malaysia was inserted with an IS981SC-IS1161 hybrid IS element. The hybrid IS element was found in all of the GCSD fish isolates and one GCSD pig through PCR screening. Although no insertion sequence (IS) was detected in the spegg locus of S. dysgalactiae ssp. equisimilis (GCSE) strains, a five-nucleotide deletion mutation was detected in the ORF of the spegg locus of one GCSE strain at the supposed site of IS981SC insertion, resulting in a frameshift mutation.

Matched MeSH terms: Streptococcus/isolation & purification
Fulltext Genetic characterization of Japanese encephalitis virus genotype II strains isolated from 1951 to 1978

Schuh AJ, Tesh RB, Barrett AD

J Gen Virol, 2011 Mar;92(Pt 3):516-27.
PMID: 21123550 DOI: 10.1099/vir.0.027110-0

Japanese encephalitis virus (JEV), the prototype member of the JEV serocomplex, genus Flavivirus, family Flaviviridae, is the most significant arthropod-borne encephalitis worldwide in terms of morbidity and mortality. At least four genotypes (GI-GIV) of the virus have been identified; however, to date, the genomic nucleotide sequence of only one GII virus has been determined (FU strain, Australia, 1995). This study sequenced three additional GII strains of JEV isolated between 1951 and 1978 in Korea, Malaysia and Indonesia, respectively, and compared them with the FU strain, as well as with virus strains representing the other three genotypes. Based on nucleotide and amino acid composition, the genotype II strains were the most similar to GI strains; however, these two genotypes are epidemiologically distinct. Selection analyses revealed that the strains utilized in this study are under predominantly purifying selection, and evidence of positive selection was detected at aa 24 of the NS4B protein, a protein that functions as an alpha/beta interferon signalling inhibitor.

Matched MeSH terms: Encephalitis Virus, Japanese/isolation & purification
Fulltext Molecular characterization and epidemiology of rotavirus isolates obtained from children with diarrhoea in Malaysia

Zuridah H, Kirkwood CD, Bishop RF, Bogdanovic-Sakran N, Yap KL

Med J Malaysia, 2009 Sep;64(3):193-6.
PMID: 20527266 MyJurnal

This retrospective study examined the G/P type of rotavirus in RNA samples that have previously been e-typed by RNA-PAGE in 1996. The results were then compared to 2007 samples to ascertain the extent of changes that may have occurred in this 11-years time interval. The G and P genotypes were determined by hemi-nested PCR and further analysed by phylogenetic study. In 1996, the G/P combination G1P[8], G(UT)P[8] and G1P(UT) prevalence rate were 81%, 9% and 7%, respectively. As expected, the G9 genotype which has already emerged worldwide was identified in 42% of the 2007 samples with the remaining 33% G1P[8] and 25% G1P(UT) Analysis of the RNA pattern showed that majority of the isolates were long e-type in both series, nevertheless minor differences within electropherotypes were observed. Genetic diversity in some strains of the human group A rotaviruses was analysed by phylogenetic methods. These findings will help in the decision to introduce rotavirus vaccines within the next decade.

Matched MeSH terms: Rotavirus/isolation & purification*
Hot compressed water pretreatment of oil palm fronds to enhance glucose recovery for production of second generation bio-ethanol

Goh CS, Lee KT, Bhatia S

Bioresour Technol, 2010 Oct;101(19):7362-7.
PMID: 20471249 DOI: 10.1016/j.biortech.2010.04.048

This work presents the pretreatment of oil palm fronds (OPF) using hot compressed water (HCW) to enhance sugar recovery in enzymatic hydrolysis. A central, composite rotatable design was used to optimize the effect of reaction temperature, reaction time and liquid-solid ratio on the pretreatment process. All variables were found to significantly affect the glucose yield. A quadratic polynomial equation was used to model glucose yield by multiple regression analysis, using response surface methodology (RSM). Using a 10 bar pressurized reactor, the optimum conditions for pretreatment of OPF were found at 178 degrees C, 11.1 min and a liquid-solid ratio of 9.6. The predicted glucose yield was 92.78 wt.% at the optimum conditions. Experimental verification of the optimum conditions gave a glucose yield in good agreement with the estimated value of the model.

Matched MeSH terms: Glucose/isolation & purification*
Molecular diversity of fungal endophytes isolated from Garcinia mangostana and Garcinia parvifolia

Sim JH, Khoo CH, Lee LH, Cheah YK

J Microbiol Biotechnol, 2010 Apr;20(4):651-8.
PMID: 20467234

Garcinia is commonly found in Malaysia, but limited information is available regarding endophytic fungi associated with this plant. In this study, 24 endophytic fungi were successfully recovered from different parts of two Garcinia species. Characterization of endophytic fungi was performed based on the conserved internal transcribed spacer (ITS) region sequence analysis and the antimicrobial properties. Results revealed that fruits of the plant appeared to be the highest inhabitation site (38 %) as compared with others. Glomerella sp., Guignardia sp., and Phomopsis sp. appeared to be the predominant endophytic fungi group in Garcinia mangostana and Garcinia parvifolia. Phylogenetic relationships of the isolated endophytic fungi were estimated from the sequences of the ITS region. On the other hand, antibacterial screening showed 11 of the isolates possessed positive response towards pathogenic and nonpathogenic bacteria. However, there was no direct association between certain antibacterial properties with the specific genus observed.

Matched MeSH terms: Ascomycota/isolation & purification
Fulltext The emerging of the fifth malaria parasite (Plasmodium knowlesi): a public health concern?

Sabbatani S, Fiorino S, Manfredi R

Braz J Infect Dis, 2010 May-Jun;14(3):299-309.
PMID: 20835518

After examining the most recent scientific evidences, which assessed the role of some malaria plasmodia that have monkeys as natural reservoirs, the authors focus their attention on Plasmodium knowlesi. The infective foci attributable to this last Plasmodium species have been identified during the last decade in Malaysia, in particular in the states of Sarawak and Sabah (Malaysian Borneo), and in the Pahang region (peninsular Malaysia). The significant relevance of molecular biology assays (polymerase chain reaction, or PCR, performed with specific primers for P. knowlesi), is underlined, since the traditional microscopic examination does not offer distinguishing features, especially when the differential diagnosis with Plasmodium malariae is of concern. Furthermore, Plasmodium knowlesi disease may be responsible of fatal cases, since its clinical presentation and course is more severe compared with those caused by P. malariae, paralleling a more elevated parasitemia. The most effective mosquito vector is represented by Anopheles latens; this mosquito is a parasite of both humans and monkeys. Among primates, the natural hosts are Macaca fascicularis, M. nemestina, M. inus, and Saimiri scirea. When remarking the possible severe evolution of P. knowlesi malaria, we underline the importance of an early recognition and a timely management, especially in patients who have their first onset in Western Hospitals, after journeys in Southeast Asian countries, and eventually participated in trekking excursions in the tropical forest. When malaria-like signs and symptoms are present, a timely diagnosis and treatment become crucial. In the light of its emerging epidemiological features, P. knowlesi may be added to the reknown human malaria parasites, whith includes P. vivax, P. ovale, P. malariae, and P. falciparum, as the fifth potential ethiologic agent of human malaria. Over the next few years, it will be mandatory to support an adequate surveillance and epidemiological network. In parallel with epidemiological and health care policy studies, also an accurate appraisal of the clinical features of P. knowlesi-affected patients will be strongly needed, since some preliminary experiences seem to show an increased disease severity, associated with increased parasitemia, in parallel with the progressive increase of inter-human infectious passages of this emerging Plasmodium.

Matched MeSH terms: Plasmodium knowlesi/isolation & purification*
Fulltext High-risk human papillomavirus in the oral cavity of women with cervical cancer, and their children

Saini R, Khim TP, Rahman SA, Ismail M, Tang TH

Virol J, 2010;7:131.
PMID: 20550718 DOI: 10.1186/1743-422X-7-131

Association of High-risk Human Papillomavirus (HR-HPV) with oral cancer has been established recently. Detecting these viruses in oral cavity is important to prevent oral lesions related to them. The purpose of this study was to evaluate the prevalence of HR-HPV in the oral cavity of women with cervical cancer, and their children. A total of 70 women, previously diagnosed with cervical cancer, and 46 children of these women, born by vaginal delivery only, were selected for this study. Buccal swabs were collected from their oral cavity and HPV detection was carried out using Hybrid Capture 2 high-risk HPV (HC2 HR-HPV) detection system.

Matched MeSH terms: Alphapapillomavirus/isolation & purification*
Fulltext Development and evaluation of polymerase chain reaction assay to detect Burkholderia genus and to differentiate the species in clinical specimens

Suppiah J, Thimma JS, Cheah SH, Vadivelu J

FEMS Microbiol Lett, 2010 May;306(1):9-14.
PMID: 20345378 DOI: 10.1111/j.1574-6968.2010.01923.x

Molecular-based techniques are becoming desirable as tools for identification of infectious diseases. Amongst the Burkholderia spp., there is a need to differentiate Burkholderia pseudomallei from Burkholderia cepacia, as misidentification could lead to false treatment of patients. In this study, conventional PCR assay targeting three genes was developed. Primers were designed for the amplification of Burkholderia genus-specific groEL gene, B. pseudomallei-specific mprA gene and B. cepacia-specific zmpA gene. The specificity and sensitivity of the assay was tested with 15 negative control strains and 71 Burkholderia spp. isolates including positive controls B. pseudomallei K96243 and ATCC B. cepacia strain. All B. pseudomallei strains were positive for groEL (139 bp) and mprA (162 bp), indicating a sensitivity of 100%. All B. cepacia strains produced amplicons for detection of groEL and zmpA (147 bp). Specificity using negative strains was 100%. In this study, a PCR assay specific for the detection of Burkholderia spp. and differentiation of the genus B. pseudomallei and B. cepacia was developed. The conventional assay has to be performed separately for each species due to the similar size of the PCR products amplified. This format may therefore be recommended for use as a diagnostic tool in laboratories where real-time PCR machines are not available. However, the real-time PCR was able to detect and differentiate the genus and species in single duplex assay.

Matched MeSH terms: Burkholderia/isolation & purification*
Fulltext Molybdate reduction by Pseudomonas sp. strain DRY2

Shukor MY, Ahmad SA, Nadzir MM, Abdullah MP, Shamaan NA, Syed MA

J Appl Microbiol, 2010 Jun;108(6):2050-8.
PMID: 19968732 DOI: 10.1111/j.1365-2672.2009.04604.x

To isolate and characterize a potent molybdenum-reducing bacterium.

Matched MeSH terms: Pseudomonas/isolation & purification
Prevalence of microsporidia in an indigenous Orang Asli community in Pahang, Malaysia

Lono A, Kumar GS, Chye TT

Trans R Soc Trop Med Hyg, 2010 Mar;104(3):214-8.
PMID: 19716577 DOI: 10.1016/j.trstmh.2009.07.006

Microsporidia are ubiquitous parasites thought to be closely related to fungi. Their presence in the environment means that humans are frequently exposed to infection. Stool samples were collected from 151 indigenous villagers from the eastern state of Pahang in 2005. The samples were concentrated with water-ether sedimentation, stained with modified trichrome stain and examined under oil-immersion microscopy. Thirty-two specimens (21.2%) were positive for microsporidia. Microsporidia were observed as ovoid or rounded ovoid shapes measuring approximately 1mum, with a bright pink outline containing a central or posterior vacuole. PCR amplification with specific primers on microscopy-positive specimens amplified Encephalitozoon intestinalis DNA from five of the ten specimens used.

Matched MeSH terms: Microsporidia/isolation & purification
Oral diosmectite reduces stool output and diarrhea duration in children with acute watery diarrhea

Dupont C, Foo JL, Garnier P, Moore N, Mathiex-Fortunet H, Salazar-Lindo E, et al.

Clin Gastroenterol Hepatol, 2009 Apr;7(4):456-62.
PMID: 19268266 DOI: 10.1016/j.cgh.2008.12.007

Diosmectite is a clay used to treat children with acute watery diarrhea. However, its effects on stool output reduction, the key outcome for pediatric antidiarrheal drugs, have not been shown.

Matched MeSH terms: Rotavirus/isolation & purification

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