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Journal of Microbiological Methods

Eloy Gonzales-Gustavson, Yexenia Cardenas-Youngs, Miquel Calvo, Marcelle Figueira Marques da Silva, Ayalkibet Hundesa, Inmaculada Amorós, Yolanda Moreno, Laura Moreno-Mesonero, Rosa Rosell, Lliliane Ganges, Rosa Araujo, Rosina Girones
In this study, the use of skimmed milk flocculation (SMF) to simultaneously concentrate viruses, bacteria and protozoa was evaluated. We selected strains of faecal indicator bacteria and pathogens, such as Escherichia coli and Helicobacter pylori. The viruses selected were adenovirus (HAdV 35), rotavirus (RoV SA-11), the bacteriophage MS2 and bovine viral diarrhoea virus (BVDV). The protozoa tested were Acanthamoeba, Giardia and Cryptosporidium. The mean recoveries with q(RT)PCR were 66% (HAdV 35), 24% (MS2), 28% (RoV SA-11), 15% (BVDV), 60% (E...
January 13, 2017: Journal of Microbiological Methods
C Dard, S Bailly, T Drouet, H Fricker-Hidalgo, M P Brenier-Pinchart, H Pelloux
BACKGROUND: Serological investigation of Toxoplasma gondii can answer many questions about toxoplasmosis in human pathology. Along these lines, studies on serum storage in biobanks need to be performed especially in terms of determining the impact of storage on relevance of sera analysis after freezing. This study assessed the impact of long-term sera storage on the stability of anti-Toxoplasma immunoglobulins. MATERIAL AND METHODS: The stability of anti-Toxoplasma IgG and IgM was studied in 244 and 242 sera respectively, stored at -20°C from one month to ten years...
January 13, 2017: Journal of Microbiological Methods
Pallop Tankaew, Thawatchai Singh-La, Chatchote Titaram, Veerasak Punyapornwittaya, Preeyanat Vongchan, Takuo Sawada, Nattawooti Sthitmatee
Pasteurella multocida causes haemorrhagic septicemia in livestock and wild animals, including elephants. The disease has been reported in Asian elephants in India and Sri Lanka, but to date there have been no reported cases in Thailand. ELISA or indirect hemagglutination assays (IHA) have been demonstrated to be able to detect the antibody against the disease in cattle, but no data are available for elephants. The present study reports a novel in-house indirect ELISA for antibody detection of haemorrhagic septicemia in Asian elephants, and evaluates the sensitivity and specificity of the method using a Bayesian approach...
January 12, 2017: Journal of Microbiological Methods
Yukari Matsunaga, Megumi Ando, Kosuke Izumitsu, Kazumi Suzuki, Yoichi Honda, Toshikazu Irie
Pleurotus ostreatus was transformed using the nourseothricin-resistant gene for the first time. The transformation efficiency was 1.3±0.6transformants/μg plasmid DNA. In addition, the transformation efficiency of the bialaphos-resistant gene was increased to 26.7±11.5transformants/μg plasmid DNA.
January 10, 2017: Journal of Microbiological Methods
Osamu Tsuzukibashi, Satoshi Uchibori, Taira Kobayashi, Koji Umezawa, Chiho Mashimo, Takayuki Nambu, Masanori Saito, Tomomi Hashizume-Takizawa, Tomoko Ochiai
Rothia dentocariosa and Rothia mucilaginosa which are Gram-positive bacteria are part of the normal flora in the human oral cavity and pharynx. Furthermore, Rothia aeria, which was first isolated from air samples in the Russian space station Mir, is predicted to be an oral inhabitant. Immunocompromised patients are often infected by these organisms, leading to various systemic diseases. The involvement of these organisms in oral infections has attracted little attention, and their distribution in the oral cavity has not been fully clarified because of difficulties in accurately identifying these organisms...
January 10, 2017: Journal of Microbiological Methods
Darren Greetham, David F Lappin, Ranjith Rajendran, Lindsay O'Donnell, Leighann Sherry, Gordon Ramage, Christopher Nile
Candida albicans metabolic activity in the presence and absence of acetylcholine was measured using phenotypic microarray analysis. Acetylcholine inhibited C. albicans biofilm formation by slowing metabolism independent of biofilm forming capabilities. Phenotypic microarray analysis can therefore be used for screening compound libraries for novel anti-fungal drugs and measuring antifungal resistance.
January 9, 2017: Journal of Microbiological Methods
Felix Matthias Fuchs, Adam Driks, Peter Setlow, Ralf Moeller
Bacterial biofilms cause severe problems in medicine and industry due to the high resistance to disinfectants and environmental stress of organisms within biofilms. Addressing challenges caused by biofilms requires full understanding of the underlying mechanisms for bacterial resistance and survival in biofilms. However, such work is hampered by a relative lack of systems for biofilm cultivation that are practical and reproducible. To address this problem, we developed a readily applicable method to culture Bacillus subtilis biofilms on a membrane filter...
January 7, 2017: Journal of Microbiological Methods
A M Celis, A M Vos, S Triana, C A Medina, N Escobar, S Restrepo, H A B Wösten, H de Cock
Malassezia spp. are part of the normal human and animal mycobiota but are also associated with a variety of dermatological diseases. The absence of a transformation system hampered studies to reveal mechanisms underlying the switch from the non-pathogenic to pathogenic life style. Here we describe, a highly efficient Agrobacterium-mediated genetic transformation system for Malassezia furfur and M. pachydermatis. A binary T-DNA vector with the hygromycin B phosphotransferase (hpt) selection marker and the green fluorescent protein gene (gfp) was introduced in M...
January 5, 2017: Journal of Microbiological Methods
Yanglei Yi, Oscar P Kuipers
In order to develop a method for electroporation of environmental Bacillus mycoides strains, we optimized several conditions that affect the electroporation efficiency of this bacterium. By combining the optimized conditions, the electroporation efficiency of strain EC18 was improved to (1.3±0.6)×10(5)cfu/μg DNA, which is about 10(3)-fold increase in comparison with a previously reported value. The method was further validated on various B. mycoides strains, yielding reasonable transformation efficiencies...
December 30, 2016: Journal of Microbiological Methods
E L Plummer, S M Garland, C S Bradshaw, M G Law, L A Vodstrcil, J S Hocking, C K Fairley, S N Tabrizi
We investigated the utility of quantitative PCR assays for diagnosis of bacterial vaginosis and found that while the best model utilized bacterial copy number adjusted for total bacterial load (sensitivity=98%, specificity=93%, AUC=0.95[95%CI=0.93,0.97]), adjusting for total bacterial or human cell load did not consistently increase the diagnostic performance of the assays.
December 29, 2016: Journal of Microbiological Methods
Robert S Burlage, Joshua Tillmann
Biosensors are devices which utilize both an electrical component (transducer) and a biological component to study an environment. They are typically used to examine biological structures, organisms and processes. The field of biosensors has now become so large and varied that the technology can often seem impenetrable. Yet the principles which underlie the technology are uncomplicated, even if the details of the mechanisms are elusive. In this review we confine our analysis to relatively current advancements in biosensors for the detection of whole bacterial cells...
December 28, 2016: Journal of Microbiological Methods
Léa Cabrol, Marianne Quéméneur, Benjamin Misson
Sodium azide (NaN3) was evaluated as inhibitor of microbial growth and activity in marine sediment resuspensions by monitoring the abundance of free-living and sessile bacteria using both flow cytometry and qPCR methods. Results show that 50mM of NaN3 strongly inhibits bacterial growth under natural and enriched resource conditions.
December 27, 2016: Journal of Microbiological Methods
Fernanda Almeida, Marta Inês Cazentini Medeiros, Dália Dos Prazeres Rodrigues, Marc W Allard, Juliana Pfrimer Falcão
CRISPR-multi-locus virulence sequence typing (CRISPR-MVLST) was performed to type 92 S. Typhimurium strains isolated from humans and food sources between 1983 and 2013 in Brazil and assess the suitability of this methodology comparing it with PFGE already used for subtyping the same strains. Among the 92 S. Typhimurium strains studied, we identified 25 CRISPR1 alleles, 27 CRISPR2 alleles, 2 fimH alleles and 3 sseL alleles showing that the genetic variability is much higher in the CRISPRs loci than in the virulence genes...
December 26, 2016: Journal of Microbiological Methods
Steven D Bowden, Nagendra P Palani, Igor G L Libourel
Site specific recombinases are invaluable tools in molecular biology, and are emerging as powerful recorders of cellular events in synthetic biology. We have developed a stringently controlled FLP recombinase system in Escherichia coli using an arabinose inducible promoter combined with a weak ribosome binding site.
December 24, 2016: Journal of Microbiological Methods
Whitney N Wood, Kyle D Smith, Jennifer A Ream, L Kevin Lewis
Many plasmids used for gene cloning and heterologous protein expression in Escherichia coli cells are low copy number or single copy number plasmids. The extraction of these types of plasmids from small bacterial cell cultures produces low DNA yields. In this study, we have quantitated yields of low copy and single copy number plasmid DNAs after growth of cells in four widely used broths (SB, SOC, TB, and 2xYT) and compared results to those obtained with LB, the most common E. coli cell growth medium. TB (terrific broth) consistently generated the greatest amount of plasmid DNA, in agreement with its ability to produce higher cell titers...
December 23, 2016: Journal of Microbiological Methods
M Rahnama, N Forester, K G S U Ariyawansa, C R Voisey, L J Johnson, R D Johnson, D J Fleetwood
A split-marker system for targeted gene deletion was developed for the model grass endophytic fungus Epichloë festucae. Compared to the conventional system that yields up to 25% homologous recombinants, the method resulted in 33-74% targeted deletions in E. festucae using as little as 1.5kb of targeting sequence.
December 22, 2016: Journal of Microbiological Methods
Angela Ma, Linda Chui
Three qPCR assays targeting the locus of heat resistance to identify heat resistant clinical Escherichia coli isolates are described. Of 613 isolates, 3 (0.5%) possessed the locus. The assays are a rapid, highly sensitive and specific alternative to screening by heat shock and can be used in food safety surveillance.
December 22, 2016: Journal of Microbiological Methods
Takaya Segawa, Mari Matsui, Masato Suzuki, Atsuko Tsutsui, Makoto Kuroda, Keigo Shibayama, Satowa Suzuki
The Carba NP test was developed to detect carbapenemase-producing Enterobacteriaceae, and uses imipenem as the reaction substrate. In Japan, IMP-6 metallo-β-lactamase (MBL) producers, which are usually resistant to meropenem but susceptible to imipenem, and IMP-1 MBL producers, which are usually resistant to both carbapenems are prevalent. We performed the Carba NP test with IMP-6 and IMP-1 MBL producers, and both types were detected by the Carba NP test with high sensitivity. All IMP-1 MBL producers were detected by the Carba NP test, but the minimum inhibitory concentrations (MICs) of imipenem varied from 0...
December 19, 2016: Journal of Microbiological Methods
Romain Briquaire, Rita R Colwell, Jacques Boncy, Emmanuel Rossignol, Aline Dardy, Isabelle Pandini, François Villeval, Jean-Louis Machuron, Anwar Huq, Shah Rashed, Thierry Vandevelde, Christine Rozand
Cholera is now considered to be endemic in Haiti, often with increased incidence during rainy seasons. The challenge of cholera surveillance is exacerbated by the cost of sample collection and laboratory analysis. A diagnostic tool is needed that is low cost, easy-to-use, and able to detect and quantify Vibrio cholerae accurately in water samples within 18-24h, and perform reliably in remote settings lacking laboratory infrastructure and skilled staff. The two main objectives of this study were to develop and evaluate a new culture medium embedded in a new diagnostic tool (PAD for paper based analytical device) for detecting Vibrio cholerae from water samples collected in Haiti...
December 19, 2016: Journal of Microbiological Methods
Shiping Deng, Richard Dick, Christopher Freeman, Ellen Kandeler, Michael N Weintraub
Data interpretation and comparison in enzyme assays can be challenging because of the complex nature of the environment and variations in methods employed. This letter provides an overview of common enzyme assays, the need for methods standardization, and solutions addressing some of the concerns in microplate fluorimetric assay approaches.
December 18, 2016: Journal of Microbiological Methods
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