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1.
Food Chem ; 337: 127762, 2021 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-32777563

RESUMO

Gelatin and collagen are considered halal-critical ingredients as they are typically derived from either bovine or porcine animals. Current analytical methods for determining the sources of gelatin and collagen suffer from limitations in terms of robustness and false positives in peptide matching. Thus, the aim of this study was to investigate the utility of monitoring hydroxyproline, a signature amino acid for gelatin and collagen, for identifying potentially haram foodstuffs. To determine the hydroxyproline profiles among animal- and plant-based samples, one-way univariate analysis of variance followed by pair-wise comparison was used to establish statistical significance. Multivariate chemometric analysis through principal component analysis revealed a discrete distribution pattern among 59 samples due to hydroxyproline variability. Finally, inter- and intra-laboratory comparisons demonstrated the validity and robustness of hydroxyproline determination according to ISO 17025. Thus, this preliminary identification technique will aid the identification of potentially haram foodstuffs.

2.
Int J Med Microbiol ; 310(7): 151449, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-33092697

RESUMO

Pneumococci are a common cause of severe infections, such as otitis media, pneumonia, meningitis and bacteremia. Pili are detected in a small proportion of pneumococcal population, but these structures have recently been associated with bacterial virulence in humans. Therefore, the epidemiological relationships between pneumococcal pili, serotype and antimicrobial resistance are of interest. This study aims to discuss the virulence contribution of the Streptococcus pneumoniae pili and the epidemiological relationships among the pilus genes, antimicrobial resistance trends, regional serotypes and genotypic variations. Previous reports have characterized the pneumococcal pilus islet as a clonal feature in the pneumococcal serotypes that are covered by the pneumococcal conjugate vaccine (PCV), including serotypes 19A, 19F, 23F and 7F. Many of the pneumococcal molecular epidemiology network (PMEN) clones are piliated isolates that are also strongly associated with a high frequency of multidrug resistance. Most of these piliated pneumococcal isolates belong to a few clonal complexes (CC), such as CC320, CC199, CC271, CC191 and CC156. Additional molecular epidemiology and genomic studies, particularly whole genome sequence analysis (WGS), are needed to develop an in-depth understanding of the piliated pneumococcal isolates.

3.
J Food Sci ; 83(10): 2409-2414, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30184265

RESUMO

High-quality DNA extracts are imperative for downstream applications in molecular identification. Most processed food products undergo heat treatments causing DNA degradation, which hampers application of DNA-based techniques for food authentication. Moreover, the presence of inhibitors in processed food products is also problematic, as inhibitors can impede the process of obtaining high qualities and quantities of DNA. Various approaches in DNA extraction and factors in structure and texture of various food matrices affecting DNA extraction are explained in this review.


Assuntos
DNA/isolamento & purificação , Análise de Alimentos , Contaminação de Alimentos/análise , Culinária , DNA/química , DNA/genética , Manipulação de Alimentos
4.
Acta Trop ; 188: 68-77, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30145261

RESUMO

Leptospirosis is caused by the spirochetal bacterium Leptospira of which rodents are considered the most important reservoir. This study aims to determine and characterize virulent Leptospira species among rodents and small mammals found in human settlements and recreational spots within the Hulu Langat and Gombak districts of Selangor, Malaysia; regions that frequently report probable human leptospirosis cases. Molecular analysis revealed an overall Leptospira detection rate of 14.3% among the 266 small mammals captured, and the human settlements were found to have the highest number of isolates (15.1%), followed by recreational sites (14.5%). The molecular characterization conducted based on the lipL32, secY genes and MLST revealed that the strains belonged to four different species, including; Leptospira interrogans (29; 76.3%; ST50, ST238, ST243), L. kirschneri (5; 13.15%; ST110), L. borgpetersenii (3; 8%; ST143) and L. weilii (1; 2.63%; ST242). The study revealed genotypes of circulating strains among small mammals in Malaysia, which include Leptospira locus ST110 L. kirschneri, ST 50 L. interrogans, ST143 L. borgpetersenii and ST242 L. weilii. Among the small mammals studied, 17/105 (16.2%) Rattus norvegicus, 7/59 (11.9%) of Rattus rattus, 5/24 (20.8%) of Maxomys whiteheadi, 4/18 (22.2%) of Sundamys muelleri, 2/22 (9%), Tupaia gliss, 2/16 (12.5%) Rattus tiomanicus and 1/4 (25%) of Suncus murinus carried pathogenic leptospires. The data from the present study may imply that, in addition to rodents, other small mammals also serve as maintenance hosts for Leptospira. Hence, much remains unknown about Leptospira maintenance hosts, and there is need for further investigation to ascertain the prevailing serovars of pathogenic Leptospira in Malaysia. This will assist in the development of efficient diagnostic assays with improved microscopic agglutination test (MAT) panels, and in the implementation of suitable prevention and control measures.


Assuntos
Leptospira/genética , Leptospirose/microbiologia , Testes de Aglutinação , Animais , Humanos , Leptospira/isolamento & purificação , Malásia , Mamíferos , Roedores
5.
Germs ; 8(1): 21-30, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29564245

RESUMO

Introduction: This study aims to assess the antimicrobial susceptibility profiles of Staphylococcus aureus strains isolated from university students and to determine the prevalence of constitutive and inducible clindamycin resistance, the latter being able to cause therapeutic failure due to false in vitro clindamycin susceptibility. Methods: S. aureus strains were isolated from the nasal swabs of 200 health sciences students of a Malaysian university. Twelve classes of antibiotics were used to evaluate the antimicrobial susceptibility profiles with the macrolide-lincosamide-streptogramin B (MLSB) phenotype for inducible clindamycin resistance determined by the double-diffusion test (D-test). Carriage of resistance and virulence genes was performed by PCR on S. aureus isolates that were methicillin resistant, erythromycin resistant and/or positive for the leukocidin gene, pvl (n=15). Results: Forty-nine isolates were viable and identified as S. aureus with four of the isolates characterized as methicillin-resistant S. aureus (MRSA; 2.0%). All isolates were susceptible to the antibiotics tested except for penicillin (resistance rate of 49%), erythromycin (16%), oxacillin (8%), cefoxitin (8%) and clindamycin (4%). Of the eight erythromycin-resistant isolates, iMLSB was identified in five isolates (three of which were also MRSA). The majority of the erythromycin-resistant isolates harbored the msrA gene (four iMLSB) with the remaining iMLSB isolate harboring the ermC gene. Conclusion: The presence of MRSA isolates which are also iMLSB in healthy individuals suggests that nasal carriage may play a role as a potential reservoir for the transmission of these pathogens.

6.
Malays J Med Sci ; 23(5): 65-71, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-27904427

RESUMO

BACKGROUND: Several occupational diseases of multiple origins are encountered among abattoir workers. Presence of indicator microorganisms (coliforms) on hands of workers can be used a gauge for hygienic practices. METHODS: A cross-sectional study was performed to assess the prevalence of E.coli and enterobacteriaceae among Halal abattoir workers in some government halal abattoirs of Malaysia. A total of one hundred and sixty-five hand swab samples were collected from workers of Halal abattoirs in Malaysia. The samples were subjected to microbiological analysis for characterisation and serotyping. RESULTS: The results have shown that no Escherichia coli O157:H7 was isolated on the hands of abattoir workers before and after work. However, a total prevalence of 9.7% was recorded for all samples during work. For non-O157:H7, total prevalence of 33.3% during work and 13% after work were obtained. High prevalence was recorded in sample taken during work from Tampin, Jasin and Kemaman (100% each) while low prevalence where observed in Shah Alam, Banting and Ipoh (20% each). CONCLUSIONS: Based on the findings the hygienic practices of hand washing among the workers in few locations was found to be low especially after work.

7.
J Agric Food Chem ; 64(32): 6343-54, 2016 Aug 17.
Artigo em Inglês | MEDLINE | ID: mdl-27501408

RESUMO

Beef, buffalo, and pork adulteration in the food chain is an emerging and sensitive issue. Current molecular techniques to authenticate these species depend on polymerase chain reaction (PCR) assays involving long and single targets which break down under natural decomposition and/or processing treatments. This novel multiplex polymerase chain reaction-restriction fragment length polymorphism assay targeted two different gene sites for each of the bovine, buffalo, and porcine materials. This authentication ensured better security, first through a complementation approach because it is highly unlikely that both sites will be missing under compromised states, and second through molecular fingerprints. Mitochondrial cytochrome b and ND5 genes were targeted, and all targets (73, 90, 106, 120, 138, and 146 bp) were stable under extreme boiling and autoclaving treatments. Target specificity and authenticity were ensured through cross-amplification reaction and restriction digestion of PCR products with AluI, EciI, FatI, and CviKI-1 enzymes. A survey of Malaysian frankfurter products revealed rampant substitution of beef with buffalo but purity in porcine materials.


Assuntos
Análise do Polimorfismo de Comprimento de Fragmentos Amplificados/métodos , Búfalos/genética , Bovinos/genética , Contaminação de Alimentos/análise , Produtos da Carne/análise , Reação em Cadeia da Polimerase Multiplex/métodos , Suínos/genética , Animais , Análise Discriminante , Polimorfismo de Fragmento de Restrição
8.
Artigo em Inglês | MEDLINE | ID: mdl-26458055

RESUMO

This paper describes a short-amplicon-based TaqMan probe quantitative real-time PCR (qPCR) assay for the quantitative detection of canine meat in chicken nuggets, which are very popular across the world, including Malaysia. The assay targeted a 100-bp fragment of canine cytb gene using a canine-specific primer and TaqMan probe. Specificity against 10 different animals and plants species demonstrated threshold cycles (Ct) of 16.13 ± 0.12 to 16.25 ± 0.23 for canine DNA and negative results for the others in a 40-cycle reaction. The assay was tested for the quantification of up to 0.01% canine meat in deliberately spiked chicken nuggets with 99.7% PCR efficiency and 0.995 correlation coefficient. The analysis of the actual and qPCR predicted values showed a high recovery rate (from 87% ± 28% to 112% ± 19%) with a linear regression close to unity (R(2) = 0.999). Finally, samples of three halal-branded commercial chicken nuggets collected from different Malaysian outlets were screened for canine meat, but no contamination was demonstrated.


Assuntos
DNA/análise , Cães/genética , Contaminação de Alimentos/análise , Carne/análise , Sondas de Oligonucleotídeos/genética , Reação em Cadeia da Polimerase em Tempo Real/métodos , Animais , Galinhas , DNA/genética , Sondas de Oligonucleotídeos/análise
9.
Jundishapur J Microbiol ; 8(11): e25177, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26855740

RESUMO

BACKGROUND: The sensing mechanism of glucose in Saccharomyces cerevisiae is well studied. However, such information is scarcely found in other yeast species such as Candida glabrata. OBJECTIVES: This study aimed to identify the glucose sensing pathway related genes of C. glabrata and to analyze the regulation pattern of these genes in response to different surrounding glucose concentrations through the quantitative real time polymerase chain reaction (qRT-PCR). MATERIALS AND METHODS: Phylogenetic analysis was carried out on predicted amino acid sequences of C. glabrata and S. cerevisiae to compare their degree of similarity. In addition, the growth of C. glabrata in response to different amounts of glucose (0%, 0.01%, 0.1%, 1% and 2%) was evaluated via the spot dilution assay on prepared agar medium. Besides, the SNF3 and RGT2, which act as putative glucose sensors, and the RGT1 and MIG1, which act as putative transcriptional regulators and selected downstream hexose transporters (HXTs), were analysed through qRT-PCR analysis for the gene expression level under different glucose concentrations. RESULTS: Comparative analysis of predicted amino acids in the phylogenetic tree showed high similarity between C. glabrata and S cerevisiae. Besides, C. glabrata demonstrated the capability to grow in glucose levels as low as 0.01% in the spot dilution assay. In qRT-PCR analysis, differential expressions were observed in selected genes when C. glabrata was subjected to different glucose concentrations. CONCLUSIONS: The constructed phylogenetic tree suggests the close evolutionary relationship between C. glabrata and S. cerevisiae. The capability of C. glabrata to grow in extremely low glucose environments and the differential expression of selected glucose-sensing related genes suggested the possible role of these genes in modulating the growth of C. glabrata in response to different glucose concentrations. This study helps deepen our understanding of the glucose sensing mechanism in C. glabrata and serves to provide fundamental data that may assist in unveiling this mechanism as a potential drug target.

10.
Pak J Pharm Sci ; 27(4): 831-5, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25015448

RESUMO

The present study aimed to determine the hepatoprotective activity of Dicranopteris linearis L. (family Gleicheniaceae) leaf aqueous extract (DLAE) using two models of liver injury in rats. Rats were divided into ten groups (n=6) and received dH2O (negative control), 200 mg/kg silymarin (positive control) or DLAE (50, 250 and 500 mg/kg) orally once daily for 7 consecutive days and on the 8th day subjected to the hepatotoxic induction either using carbon tetrachloride (CCl4) or paracetamol (PCM). The bloods and livers were collected and subjected to biochemical and microscopical analysis. From the data obtained, only the highest dose of DLAE significantly (P<0.05) reduced the ALP, ALT and AST levels in CCl4-and PCM-induced hepatotoxic rats while the other doses caused significant (P<0.05) reduction only in the levels of ALT and AST. The histological results obtained were in line with the biochemical analysis wherein reduction in the CCl4- and PCM-induced tissue formation of necrosis, steatosis and inflammation occurred in a dose-dependent manner. In conclusion, the DLAE possesses hepatoprotective activity, which could be attributed to its free radicals scavenging and antioxidant activities, and high flavonoids content. Thus, in-depth studies regarding the hepatoprotective activity of DLAE are warranted.


Assuntos
Acetaminofen/toxicidade , Doença Hepática Induzida por Substâncias e Drogas/prevenção & controle , Gleiquênias , Extratos Vegetais/farmacologia , Animais , Tetracloreto de Carbono/toxicidade , Doença Hepática Induzida por Substâncias e Drogas/patologia , Fígado/efeitos dos fármacos , Fígado/patologia , Masculino , Fitoterapia , Folhas de Planta , Ratos , Ratos Sprague-Dawley
11.
Genome Announc ; 2(2)2014 Apr 10.
Artigo em Inglês | MEDLINE | ID: mdl-24723714

RESUMO

Here, we report the draft genome sequence of a methicillin-resistant Staphylococcus aureus (MRSA) strain, KT/Y21, isolated from a blood sample of a pediatric patient. This strain belongs to sequence type 772 (ST772), harbors the staphylococcal cassette chromosome mec element (SCCmec) type V, and is positive for the Panton-Valentine leukocidin (PVL) pathogenic determinant.

12.
Infect Genet Evol ; 21: 391-4, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24342879

RESUMO

Streptococcus pneumoniae is an epidemiologically important bacterial pathogen. Recently, we reported the antibiotic susceptibility patterns of a limited collection of pneumococcal isolates in Malaysia with a high prevalence of erythromycin resistant strains. In the present study, 55 of the pneumococcal isolates of serotype 19F were further analysed by pulsed field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). The generated genotypic patterns were then correlated with the antibiograms previously reported. Forty-seven different PFGE profiles (PTs) were obtained, showing that the isolates were genetically diverse. MLST identified 16 sequence types (STs) with ST-236 being predominant (58.2%), followed by ST-81 (10.3%). Among the ST-236 isolates, 22 were erythromycin resistant S. pneumoniae (ERSP) and 15 were trimethoprim/sulfamethoxazole (TMP/SMX) resistant, while among ST-81, four isolates were ERSP and two were TMP/SMX resistant. The high prevalence of erythromycin resistant serotype 19F isolates of ST-236 in this study has also been reported in other North and South East Asian countries.


Assuntos
Farmacorresistência Bacteriana Múltipla/genética , Infecções Pneumocócicas/epidemiologia , Infecções Pneumocócicas/microbiologia , Streptococcus pneumoniae/genética , Antibacterianos/farmacologia , DNA Bacteriano/análise , Eletroforese em Gel de Campo Pulsado , Genótipo , Humanos , Malásia/epidemiologia , Tipagem de Sequências Multilocus , Filogenia , Streptococcus pneumoniae/efeitos dos fármacos , Streptococcus pneumoniae/isolamento & purificação
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