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Journal of Microbiological Methods

Nelly Sophie Raymond, Dorette Müller Stöver, Lars Stoumann Jensen, Sebastian Håkansson
The impact of formulation and desiccation on the shelf life of phosphate (P)-solubilising microorganisms is often under-studied, particularly relating to their ability to recover P-solubilisation activity. Here, Penicilllium bilaiae and Aspergillus niger were formulated on vermiculite (V) alone, or with the addition of protectants (skimmed milk (V + SM) and trehalose (V + T)), and on sewage sludge ash with (A + N) and without nutrients (A), and dried in a convective air dryer. After drying, the spore viability of P...
May 17, 2018: Journal of Microbiological Methods
Yalong Yang, Kher Zahr, Michael W Harding, Stephen E Strelkov, Krista Zuzak, David Feindel, Jie Feng
A commonly used protocol for DNA extraction from Plasmodiophora brassicae was modified by adding an alkaline treatment step to increase the purity of resting spores. The quality of DNA extracted by the modified protocol was improved due to the removal of DNA contamination from host plant cells and other microorganisms.
May 16, 2018: Journal of Microbiological Methods
Claudia D Norzagaray-Valenzuela, Lourdes J Germán-Báez, Marco A Valdez-Flores, Sergio Hernández-Verdugo, Luke M Shelton, Angel Valdez-Ortiz
Microalgae are photosynthetic microorganisms widely used for the production of highly valued compounds, and recently they have been shown to be promising as a system for the heterologous expression of proteins. Several transformation methods have been successfully developed, from which the Agrobacterium tumefaciens-mediated method remains the most promising. However, microalgae transformation efficiency by A. tumefaciens is shown to vary depending on several transformation conditions. The present study aimed to establish an efficient genetic transformation system in the green microalgae Dunaliella tertiolecta using the A...
May 16, 2018: Journal of Microbiological Methods
Javier Campos-Gomez, Jorge A Benitez
RNA polymerase containing the stress response regulator σS subunit (RpoS) plays a key role in bacterial survival in hostile environments in nature and during infection. Here we devise and validate a simple cell-based high throughput luminescence assay for this holoenzyme suitable for screening large chemical libraries in a robotic platform.
May 12, 2018: Journal of Microbiological Methods
Ping Chen, S L Rajasekhar Karna, Kai P Leung
Deletion and insertion clonal complex 8 Staphylococcus aureus mutants were created without using intermediate host S. aureus RN4220 or temperature-sensitive shuttle vectors. These mutants were created using a common cloning vector by passing the constructs through a modification host and recovering the electroporated cells in a large volume of medium.
May 11, 2018: Journal of Microbiological Methods
Jungho Kim, Sungyoung Bong, Deok-Ryun Kim, Sang-Nae Cho, Hyejon Lee
Much effort has been made to reduce the cost of LTBI diagnosis with equivalent efficacy and efficiency of interferon-γ release assay (IGRA). This study showed that repeatability, intermediate precision, and accuracy of the Bionote-ELISA were comparable to QFT-ELISA. The Bionote-ELISA could provide the alternative method.
May 11, 2018: Journal of Microbiological Methods
E F van Ess, S Ouburg, J A Land, S A Morré
OBJECTIVES: Chlamydia trachomatis (CT) IgG serology is used in many fertility clinics in order to estimate the risk for tubal factor infertility (TFI) in the fertility work-up. The predictive value for TFI of the currently used mono-target CT serology test should be improved. This study compares the performance of the new multi-target Mikrogen recomWell CT IgG ELISA with the Mikrogen recomLine CT immunoblot and visualizes distribution of individual antibodies in serum with the immunoblot in order to potentially improve the current CT IgG serology test that is clinically used...
May 7, 2018: Journal of Microbiological Methods
Prakit Saingam, Bo Li, Tao Yan
DNA-based molecular detection of microbial pathogens in complex environments is still plagued by sensitivity, specificity and robustness issues. We propose to address these issues by viewing them as inadvertent consequences of requiring specific and adequate amplification (SAA) of target DNA molecules by current PCR methods. Using the invA gene of Salmonella as the model system, we investigated if next generation sequencing (NGS) can be used to directly detect target sequences in false-negative PCR reaction (PCR-NGS) in order to remove the SAA requirement from PCR...
May 7, 2018: Journal of Microbiological Methods
Suresh Sah, Priyanka Bordoloi, D Vijaya, Satish Kumar Amarnath, C Sheela Devi, V A Indumathi, K Prashanth
Coagulase-negative staphylococci (CoNS) have been increasingly recognized as a clinically important group of species that can cause several opportunistic nosocomial infections. There are at least 47 known species of Staphylococci and to differentiate all these species >40 biochemical tests need to be performed. The present study was able to refine the CoNS identification process by using only five tests to identify S. epidermidis from the rest and used six other tests to identify eleven other clinically significant CoNS species...
May 3, 2018: Journal of Microbiological Methods
Clémence Massip, Hélène Guet-Revillet, Marion Grare, Agnès Sommet, Damien Dubois
Modified Cary-Blair medium derived devices have been implemented in many laboratories to optimize culture recovery of common bacterial enteric pathogens. Our analysis constitutes the first report of routine laboratory experience supporting the idea that the use of such devices enhances Campylobacter recovery from stools.
May 3, 2018: Journal of Microbiological Methods
Yi Wang, Weiqiang Yan, Yan Wang, Jianguo Xu, Changyun Ye
Klebsiella pneumoniae (K. pneumoniae), as an important hospital-acquired bacterium, is responsible for severe morbidity and mortality among the elderly, newborn and immune-compromised people. We established a rcsA gene-based label-free multiple cross displacement amplification (MCDA) assay for rapid, simple and sensitive detection of K. pneumoniae by using lateral flow biosensor (LFB). MCDA reaction was conducted at a fixed temperature (65 °C) for only 30 min, and amplification results were directly indicated using LFB...
May 3, 2018: Journal of Microbiological Methods
Prajakta P Kamble, Maheshkumar V Kore, Sushama A Patil, Jyoti P Jadhav, Yasmin C Attar
Tithonia rotundifolia is an easily available and abundant inulin rich weed reported to be competitive and allelopathic. This weed inulin is hydrolyzed by inulinase into fructose. Response surface methodology was employed to optimize culture conditions for the inulinase production from Arthrobacter mysorens strain no.1 isolated from rhizospheric area of Tithonia weed. Initially, Plackett- Burman design was used for screening 11 nutritional parameters for inulinase production including inulin containing weeds as cost effective substrate...
May 3, 2018: Journal of Microbiological Methods
Jiaying Sun, Ruidi Xu, Shuqin Xiao, Yuanyuan Lu, Qifeng Zhang, Chunsheng Xue
Agrobacterium tumefaciens-mediated transformation (ATMT) has been widely used in filamentous fungi. In this study, an efficient Agrobacterium tumefaciens-mediated transformation approach was developed for the plant pathogenic fungus, Kabatiella zeae, the causative pathogen of eyespot in maize. Five parameters were selected to optimize efficiencies of transformation. A. tumefaciens concentration, conidia concentration of K. zeae and mixing ratio of A. tumefaciens and K. zeae were found to exert a significant influence on all parameters...
May 3, 2018: Journal of Microbiological Methods
Ramila Cristiane Rodrigues, Anne-Lise Pocheron, Jean-Michel Cappelier, Odile Tresse, Nabila Haddad
Campylobacter jejuni is the most prevalent foodborne bacterial infection agent. This pathogen seems also involved in inflammatory bowel diseases in which pro-inflammatory cytokines, such as tumor necrosis factor α (TNFα), play a major role. C. jejuni pathogenicity has been extensively studied using in vitro cell culture methods, and more precisely "healthy" cells. In fact, no information is available regarding the behavior of C. jejuni in contact with TNFα-stimulated cells. Therefore, this research was designed to investigate the effect of TNFα on C...
May 2, 2018: Journal of Microbiological Methods
Simen Akkermans, Barbara Jaskula-Goiris, Filip Logist, Jan F Van ImpeProf
Modelling methods applied in predictive microbiology generally neglect the importance of uncertainty on the measurement of the independent variables. The Ordinary Least Squares (OLS) method that is commonly applied in predictive microbiology is only applicable if the experimental error on the inputs of the model are insignificant. However, this does not apply for many types of experimental measurements of the independent variables. Therefore, a parameter estimation method was adapted in this research for the estimation of the parameters of secondary models, taking into account uncertainty on the measurement of the influencing food characteristics...
April 25, 2018: Journal of Microbiological Methods
Michael Böhringer, Alexander Rakin, Heinrich Neubauer, Lisa D Sprague
A rapid method was developed to determine the invasion frequency of enteropathogenic Yersinia into intestinal C2BBe1 cells by means of flow cytometry. Bacteria are labelled with a thiol-cleavable amine-reactive biotin and subsequently incubated with the fluorochrome-labelled biotin-ligand neutravidin. After infection of the intestinal cells with the labelled bacteria, the neutravidin-coupled fluorochrome is detached by breaking up the linker through reduction of the disulphide. Despite reduced adhesion and invasion frequencies of the labelled bacteria into C2BBe1 cells this procedure offers the basis for the development of a fast single-step staining protocol for the recovery of invading bacteria in in a host-pathogen system for further transcriptome or proteome analysis...
April 23, 2018: Journal of Microbiological Methods
Peng Chang, Wenjuan Wang, Yasuo Igarashi, Feng Luo, Jiangye Chen
Candida albicans is an opportunistic pathogenic fungus which causes superficial and systemic infections in immunocompromised patients. It is important to characterize the roles of genes involved in its pathogenesis, virulence, and drug resistance. Several genetic manipulation toolkits have been developed for gene function research in C. albicans. Here, we describe efficient vector systems that allow economical and rapid C-terminal and N-terminal epitope-tagging, inducible and constitutive promoter replacements, and ectopic gene overexpression in C...
April 23, 2018: Journal of Microbiological Methods
Juan Gabriel Costa, María Julia Vilariño
In this work we present a novel methodology to differentiate the phases of toxoplasmosis infection: the "semiquantitative Dot Blot". It is a simple technique that does not require expensive equipment, does not involve a long technique development, and can be used in a low-complexity laboratory. In this study, two recombinant sequences of Toxoplasma gondii GRA8 antigen were used, and specific IgG antibodies were detected in selected patient samples. This method makes it possible to obtain a score for each serum and define whether the patient is in the acute or chronic phase of the infection...
April 20, 2018: Journal of Microbiological Methods
Nitu Thakore, Steve Garber, Arial Bueno, Peter Qu, Ryan Norville, Michael Villanueva, Darrell P Chandler, Rebecca Holmberg, Christopher G Cooney
Systems that automate extraction of nucleic acid from cells or viruses in complex clinical matrices have tremendous value even in the absence of an integrated downstream detector. We describe our bench-top automated workstation that integrates our previously-reported extraction method - TruTip - with our newly-developed mechanical lysis method. This is the first report of this method for homogenizing viscous and heterogeneous samples and lysing difficult-to-disrupt cells using "MagVor": a rotating magnet that rotates a miniature stir disk amidst glass beads confined inside of a disposable tube...
April 17, 2018: Journal of Microbiological Methods
Liangcheng Jiao, Qinghua Zhou, Wu Liu, Yunjun Yan
Posttransformational vector amplification (PTVA) is widely used to enrich the gene-copy number in Pichia pastoris. We engineered two test strains for PTVA studies and demonstrate that the PTVA process results in the amplification of a fragment with the resistant gene flanked by two homologous arms instead of the entire vector.
April 17, 2018: Journal of Microbiological Methods
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