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Zhenbo Xu, Lin Li, Xihong Zhao*, Jin Chu, Bing Li, Lei Shi, Jianyu Su* and Mark E. Shirtliff,
Abstract: In this study, a novel multiplex- polymerase chain reaction (PCR) for rapid detection of various staphylococci strains, including methicillin-resistant Staphylococcus aureus (MRSA), methicillin-sensitive Staphylococcus aureus, (MSSA) methicillin-resistant coagulase-negative staphylococci (MRCNS), methicillin-sensitive coagulase-negative staphylococci (MSCNS) and non-staphylococci strains, had been developed and applied. Six primers were specially designed on three target genes, which were mecA, 16S Ribosomal ribonucleic acid (rRNA) and femA. The specific amplification generated 3 bands on agarose gel, with sizes 374 bp for mecA, 542 bp for 16S rRNA and 823 bp for femA, respectively. The PCR product showed highest levels of resolution of DNA when 250 M of dNTP, primer concentration of mecA, 16S rRNA and femA reaching 1, 1 and 3 M respectively. No false positive amplification was observed, indicating the high specificity of the established multiplex PCR assay. Application of this multiplex-PCR had been further performed on detection for 262 MRSA and MRCNS strains with primers pairs M1 with M2 and F1 and F2. According to the results, multiplex-PCR results showed expected products for either MRSA or MRCNS strains, demonstrating the multiplex-PCR assays established in this study to be useful and powerful methods for differentiation of MRSA, MSSA, MRCNS, MSCNS and non-staphylococci strains.[...] Read More.
Keywords: Staphylococcus, multiplex-PCR, rapid detection.
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文章
Eisa Tahmasbpour Marzony, Mahdi Kamali, Mojtaba Saadati, Amir Homayoun Keihan*, Abbas Ali Imani Fooladi , and Sharareh Sajjadi
Abstract: Strains of shiga toxin–producing Escherichia coli (STEC) and Shigella dysenteriae type1 have been associated with outbreaks of diarrhea, hemorrhagic colitis and hemolytic uremic syndrome (HUS) in humans. Most clinical signs of disease arise as consequence of the production of shiga toxin/shiga toxin 1 (Stx/Stx1), shiga toxin 2 (Stx2) or combination of these toxins. Here, we designed a Multiplex PCR technique to identify stx/stx1 and stx2 genes with the incorporation of mdh gene of E. coli and Shigella. A total of six primers were used: SFI and SRI, which produce a 199bp product that serves as an internal positive control; Ka2F and Ka2R, which yield a 381bp fragment of stx2 gene, and Ka1F and Ka1R, which amplify a 622bp fragment of stx/stx1. The thermal profile, which was preceded by a 5 min incubation at 95°C for 20 to 25 cycles with the following parameters: 95°C at 1 min, 60°C at 1 min, 72°C at 1 min, and 5 min incubation at 72°C as final extension. PCR amplification products identifying the stx/stx1 and stx2 gene sequences were observed only in E. coli 0157:H7 and Shigella dysenteriae type1. Template nucleic acid extracted from other Gram-negative bacteria was found to be negative. The sensitivity of the PCR procedure for detection of shiga toxin genes was determined to be 2.1 pg/µl of total nucleic acid and 320 cfu/µl.[...] Read More.
Keywords: Shiga toxin, multiplex PCR, diagnosis.
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文章
HaiKuan Wang, AnDong Li , FeiFei Liu and Wei Qi*
Abstract: Clostridium butyricum is a widely utilized probiotics, also used as an alternative to antibiotics for humans and growth promotion in a wide variety of livestock species. The objective of this study is to develop an economical and practical fermentation medium for the growth of C. butyricum TK2 using orthogonal test. The optimal fermentation medium was obtained by the single factor and the orthogonal test of carbon source, nitrogen source and growth factors design. The fermentation medium was estimated to be the most economical formula (per liter): 8 g glucose, 20 g soybean meal (hydrolyzed for 3 h by neutral protease), 5 g brewer’s yeast powder. After incubating for 48 h in the optimum fermentation medium, the populations of C. butyricum TK2 were estimated to be 8 × 108 CFU ml-1, while the cost was 90% lower than ever. The optimized medium is not only more economical but also good for the growth of C. butyricum TK2. The low cost medium developed in this study can be used for large-scale commercial application where economics are quite likely to be important.[...] Read More.
Keywords: Clostridium butyricum, soybean meal, brewer’s yeast powder, medium optimization.
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Hsiu Fen Lin, Tsang Hai Chen, and Shan Da Liu*
Abstract: The inhibition zones of three Bacillus subtilis isolates, BS-99, BS-23857 and BS-33608 against Pestalotiopsis eugeniae were, 13.5, 0, and 0 mm, respectively. BS-99 showed the strongest inhibitory activity, whereas, no inhibitory activity was reported for BS-23857 or BS-33608. The spore germination rates of P. eugeniae were not significantly inhibited by n-hexane, ethyl acetate (EtOAc), or methanol (MeOH) extracts of the fermentation broths of BS-99, BS-23857, and BS-33608. However, the hyphae of P. eugeniae became swollen and malformed after 12 h and stopped growing after 30 h of treatment with the MeOH extract of the BS-99 fermentation broth. Analysis by polymerase chain reaction (PCR), blood agar test plates, and high performance liquid chromatography (HPLC) indicated that, BS-99 produced the antibiotics, iturin A and surfactin, whereas, BS-23857 and BS-33608 only produced surfactin. These data suggest that, the antifungal activity of the B. subtilis BS-99 isolate against P. eugeniae is activated only when both iturin A and surfactin are present and not by surfactin alone. Unveiling the antifungal mechanism of B. subtilis, BS-99 could promote its commercial development and application as a biofungicide for controlling P. eugeniae infections of the wax apple, which is a highly valued fruit in Taiwan.[...] Read More.
Keywords: Bacillus subtilis, iturin A, solvent extract, surfactin.
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A. Maripandi,*, Arun Kumar T. and Ali A. Al Salamah
Abstract: Dental caries is a microbiologic infection of the tooth which results in localized dissolution and destruction of the calcified tissue and it is one of the most common chronic infectious diseases in humans throughout the world. People spent huge amounts of money and time in treating dental caries. Hence, it is essential to prevent and control dental caries in public health. In our studies, a total of 70 clinical specimens, 37 from male and 33 from female dental caries patient’s samples were collected. All the clinical samples showed positive results with 87 bacterial isolates from which 63 were facultative anaerobes and 24 were anaerobe, among facultative anaerobe, Streptococcus mutants (22.98%) S. salivarius (13.79%) and Candida spp (14.97%), in anaerobes, Prevotella spp (12.64%) and Fusobacterium spp (5.74%). The antibiotic susceptibility test was carried out against Streptococcus spp. the results showed all isolates were resistance to penicillin and sensitive to tetracycline. The antibacterial activity of commercially available tooth paste was checked against Streptococcus spp., by agar well diffusion method. The results showed that pepsodent and colgate have good antibacterial activity against Streptococcus spp.[...] Read More.
Keywords: Dental caries, Streptococcus spp, Prevotella spp, antibiotics, tooth pastes.
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文章
Full Length Researc
Abstract: Diesel biodegradation and the effect of application frequency were investigated in artificially contaminated soils (loam soil and sea sand) amended with commercial fertilizers and the effect of application frequency. Two sets of four equal portions (2 kg each) of diesel-contaminated soils were amended with 10% (w/w) of 2 different commercial fertilizers at 4 different but regular intervals for up to four times during a 7- week study period. The microcosms were then incubated at 30°C and sampled weekly. One sterilized soil microcosm contaminated with 10% (v/w) diesel and supplemented with 10% (w/w) fertilizer and one naturally attenuated microcosm served as controls. The amount of diesel remaining in each sample was determined using the Soxhlet extraction method and compared to the amount of diesel present in the same sample at day 0. The quantities of hydrocarbon contents in the supplemented samples and the controls were determined by GC-MS. The total heterotrophic population of each sample was also monitored. The population increase was found to correspond to the higher diesel degradation percentage in the study on sea sand. A significant enhancement (p[...] Read More.
Keywords: Diesel, bioremediation, fertilizers, application frequency.
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Guihong Lan, Zengting Li, Hui zhang, Changjun Zou, Dairong Qiao and Yi Cao*
Abstract: Thermophilic microbial diversity in production water from a high temperature, water-flooded petroleum reservoir of an offshore oilfield in China was characterized by enrichment and polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) analysis. Six different function enrichment cultures were cultivated one year, at 75°C DGGE and sequence analyses of 16S rRNA gene fragments were used to assess the thermophilic microbial diversity. A total of 27 bacterial and 9 archaeal DGGE bands were excised and sequenced. Phylogenetic analysis of these sequences indicated that 21 bacterial and 7 archaeal phylotypes were affiliated with thermophilic microbe. The bacterial sequences were mainly bonged to the genera Fervidobacterium, Thermotoga, Dictyoglomus, Symbiobacterium, Moorella, Thermoanaerobacter, Desulfotomaculum, Thermosyntropha, Coprothermobacter, Caloramator, Thermacetogenium, and the archaeal phylogypes were represented in the genera Geoglobus and Thermococcus, Methanomethylovorans, Methanothermobacter Methanoculleus and Methanosaeta. So many thermophiles were detected suggesting that they might be common habitants in high-temperature petroleum reservoirs. The results of this work provide further insight into the thermophilic composition of microbial communities in high temperature petroleum reservoirs.[...] Read More.
Keywords: Thermophilic microbial diversity, petroleum reservoir, denaturing gradient gel electrophoresis.
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文章
Nermeen A. ElSersy* and Eman A. H. Mohamed
Abstract: A polyphasic approach was used to discriminate among the significantly hemolytic and closely related Bacillus licheniformis strains, NE-1, NE-2 and NE-3. Utilization of the API 20E kit revealed that B. licheniformis NE-1 is unique in its ability to utilize sorbitol as a sole carbon source, whereas B. licheniformis NE-2 is an exceptional in L-arabinose utilization. The phylogenetic analysis using the 16S rDNA 5´end revealed a strong relatedness to B. licheniformis strains found in the GenBank with a similarity percentage ranging from 97 to 99. Moreover, the partial sequences was successfully differentiated among the novel strains. The plasmid pattern is almost the same for the strains and plasmid curing using acridine orange showed no effect on the hemolytic activity of them.[...] Read More.
Keywords: Bacillus licheniformis, aquacultures, hemolytic activity, plasmids, 16S rDNA.
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文章
Eissa Piri, Mahdi Babaeian*, Abolfazl Tavassoli and Yasser Esmaeilian
Abstract: Global change will definitely introduce changes in agricultural ecosystems that will affect plant productivity. However, the effects on plants will be different for each region depending on the pre-existing climatic conditions and the adaptation potential of local cultivated species. The solar energy from the sun is essential to support the life on our plant, via the process of photosynthesis. However a small proportion of solar spectrum contains short-wavelength ultraviolet-B light (280 to 320 nm), which is deleterious to life. The depletion of stratospheric ozone layer by manmade pollution has substantially increased UV-B light impinging on the earth surface. UV-B affects living organisms by damaging cellular metabolism in several ways, such as dimmers formation in the genetic material DNA, disruption of membrane structure, inactivation of enzymes and generation of highly reactive free radicals. Elevated UV exposure also causes temporary or irreversible damage to the process of photosynthesis. Therefore, increased UV-B radiation would affect the stability of ecosystems and genetic health of living organisms. Many species of plants have evolved mechanisms for protection against deleterious effects of UV-B radiation. Accumulation of the UV-B absorbing pigments such as flavonoids is one of the ways by which plants alleviate the harmful effects of UV-B light.[...] Read More.
Keywords: Global change, agriculture, UV radiation, absorbing compounds.
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文章
Hamit Kavak* and Erkan Boydak
Abstract: In this work, sudden wilt syndrome in sesame was investigated during varying irrigation treatments. For the four irrigation intervals numbers of infected plants and infected rows were determined in experiments conducted during growing periods. The resulting data were transformed to percent values and regressed against the irrigation intervals. The correlations between the rates of infected plants and irrigation intervals and between the rates of infected rows and irrigation intervals were assessed separately in replicate and total plots following testing. Positive increases were observed for both categories within total plots. The correlations for the two years were r2 = 0.86 and r2 = 0.85 for the wilted plant category, and r2 = 0.99 and r2 = 1 for the infected row category. Fusariım oxysporum fsp sesame and Macrophomina phaseolina were two of the parasitic pathogens isolated from samples. In addition, parasitic pathogens were not isolated from some of the samples that displayed sudden wilt symptoms. Depending on delays in irrigation within irrigated crops grown in drought and hot regions, sudden wilt syndrome of sesame may increase. The causes of sudden wilt syndrome appear to be parasitic pathogens, drought stress, or the combined effect of both.[...] Read More.
Keywords: Sesame, irrigation intervals, sudden wilt.
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