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Journal of Microbiological Methods

I Cornet, N Wittner, G Tofani, S Tavernier
Since the determination of the fermentation kinetics is one of the main challenges in solid state fermentation, the quantitative measurement of biomass growth during microbial pretreatment by FTIR spectroscopy in Attenuated Total Reflectance mode was evaluated. Peaks at wave numbers of 1651 cm-1 and 1593 cm-1 showed to be affected during pretreatment of poplar wood particles by Phanerochaete chrysosporium MUCL 19343. Samples with different microbial biomass fractions were obtained from two different experiments, i...
January 12, 2018: Journal of Microbiological Methods
A M Gaudreau, J Labrie, C Goetz, S Dufour, M Jacques
We evaluated MALDI-TOF mass spectrometry to identify bacteria from biofilms. We compared three sample preparation procedures on biofilms grown in vitro. The extended direct transfer method was able to identify 13 isolates out of 18 (72%) at the species level and 15 out of 18 (83%) at the genus level.
January 10, 2018: Journal of Microbiological Methods
Flávia Fernandes de Mendonça Uchôa, Adriana Pittella Sudré, Sabrina Destri Emmerick Campos, Nádia Regina Pereira Almosny
Enteric parasitic diseases including giardiasis are of public health concern. Different methods are available for the diagnosis of this parasitic infection in fecal samples such as the identification of protozoan cysts and trophozoites by light microscopy, detection of specific antigens by ELISA, and amplification of DNA fragments by PCR. The present study aimed at assessing the performance of four laboratory tests for the detection of Giardia duodenalis in fecal specimens from three different host species with a previous diagnosis of giardiasis; canine, feline and human patients provided new stool samples to be retested for Giardia before initiating treatment with antiprotozoal drugs...
January 9, 2018: Journal of Microbiological Methods
Qian Zhang, Mengge Liu, Shiyu Li, Zhenrui Xu, Jiajia Wang, Yuzhe Wang, Zhengbin Fei, Wenhua Huang, Hanxiao Sun
Bifidobacterium has been developed for the oral delivery of peptides and has the added beneficial effect on our bodies through its probiotic properties. Here, we utilize Bifidobacterium as a delivery system to orally deliver Glucagon like peptide-2 (GLP-2). We constructed vector derived from pET-31b(+) to construct a Bifidobacterium longum expressing GLP-2. We then determined the bioactivity of recombinant Bifidobacterium in Caco-2 cells. Finally, we quantified newly synthesized ApoB48 and chylomicron production in mice infused with exogenous GLP-2 or Bifidobacterium expressing GLP-2...
January 9, 2018: Journal of Microbiological Methods
Takahiro Nomura, Yusuke Hashimoto, Jun Kurushima, Hidetada Hirakawa, Koichi Tanimoto, Zheng Bo, Genjie Ruan, Feng Xue, Jian Liu, Junzo Hisatsune, Motoyuki Sugai, Haruyoshi Tomita
New colony multiplex PCR assays for detection of seven types of vancomycin-resistance determinants and eight types of Enterococcus species were developed. For 135 enterococcal isolates examined in this study, these assays showed high sensitivity and specificity, and could provide the rapid and accurate detection of vancomycin-resistant determinants and Enterococcus spp.
January 5, 2018: Journal of Microbiological Methods
G Miltgen, P Plésiat, A Mille, P Chatelain, D Fournier
A new MALDI-TOF MS imipenem hydrolysis assay was developed to optimize detection of carbapenemase-producing strains of Pseudomonas aeruginosa. The method correctly discriminated carbapenemase producers (n=74) from non-producers (n=95) in 99.4% of cases, and successfully detected 100% GES-5 strains (n=5).
January 4, 2018: Journal of Microbiological Methods
Yun-Chen Lee, Ching-Fang Chien, Nai-Chun Lin
Recent advances in next generation sequencing technology allow us to retrieve the whole genome sequence of a requested bacterium in less than a day. Thus, development of quick, easy and efficient means to systemically analyze the functions of all genes is required in the post-genome era. Here, a procedure of finding a suitable chromosome integration site and developing a gene disruption system into a knock-in system in Gram-negative bacteria is proposed. As a proof of concept, we successfully modified a sacB-based gene knock-out strategy into a site-specific gene integration system to deliver a DNA fragment into the genome site between 313,520bp and 313,521bp of the model phytopathogenic bacterium, Pseudomonas syringae pv...
January 3, 2018: Journal of Microbiological Methods
Judit Serra-Pladevall, Carlos Gulin Blanco, Neus Vila Olmo, Pilar Arjona Camacho, Andreu Domingo Antonia
Although molecular diagnosis has replaced culture as the reference technique in gonococcal infection, conventional culture is still performed. The aim was to compare the survival of Neisseria gonorrhoeae (NG) in transport swabs at room temperature (RT) and at 4°C. Fifty-four NG isolates were tested. A 1.5×108CFUmL-1 suspension was prepared for each strain, in which two swabs (Amies liquid medium with flocked swabs) were absorbed; one was preserved at RT and the other at 4°C. At 24 and 48h, 150μL of Amies medium were plated in Thayer Martin agar...
December 29, 2017: Journal of Microbiological Methods
Henry P Mishek, Stephen A Stock, Jacqueline D Florick, Wilson R Blomberg, Josef D Franke
We experimentally determined minimal media requirements for Gemmata obscuriglobus, a Gram-negative Planctomycete bacteria with several unusual physiological features. We find that supplementing media with the usual vitamins solution does not improve viability, but does result in an increased growth rate in liquid cultures and a larger colony size on agar plates. By systematically including individual vitamins, or omitting individual vitamins, from media we find that the addition of only two vitamins, biotin and cyanocobalamin, are sufficient to restore colony growth to comparable rates as other commonly used media...
December 29, 2017: Journal of Microbiological Methods
David L Evers, Franz A Frye, Peter J Weina
This work developed a simple empirical algorithm to distinguish three Leishmania species using MALDI-TOF mass spectrometry. It suggests that complicated computer algorithms may not always be necessary for clinically useful microbiology applications.
December 28, 2017: Journal of Microbiological Methods
Tadayuki Iwase, Chiaki Okai, Yuko Kamata, Akiko Tajima, Yoshimitsu Mizunoe
Cellular glycogen levels reflect the activity of RpoS, an important stress-inducible bacterial sigma factor known to regulate several stress-resistance related genes, such as katE, encoding hydroperoxidase II (HPII), and the glg genes, encoding glycogen synthesis enzymes, in Escherichia coli. In this study, a straightforward assay for measuring glycogen levels and RpoS activity was developed combining the ease and simplicity of qualitative approaches. The assay reagent was a 2% iodine solution (2% iodine/1M NaOH), and the basic principle of this assay is the iodine-glycogen reaction, which produces a reddish brown color that can be measured using a spectrophotometer...
December 27, 2017: Journal of Microbiological Methods
Xi-Jun Tang, Ze Yang, Xin-Bin Chen, Wen-Fang Tian, Cheng-Ning Tu, Hai-Bo Wang
Salmonella spp./Shigella spp. are often associated with food poisoning and fecal-oral transmission of acute gastroenteritis that requires strict monitoring, especially among people who would handle food and water. In 2014, the National Health and Family Planning Commission of the P. R. China issued a national standard protocol (recommendatory) for the screening of Salmonella spp./Shigella spp.. However, its performance has not been fully studied. Whether it was suitable for use in our laboratory was still unknown...
December 17, 2017: Journal of Microbiological Methods
Hiromi Yamakawa, Kosuke Kosai, Yasuhide Kawamoto, Norihiko Akamatsu, Junichi Matsuda, Norihito Kaku, Naoki Uno, Yoshitomo Morinaga, Hiroo Hasegawa, Katsunori Yanagihara
BD Phoenix™ is an automated bacterial identification and susceptibility testing system. Here, its performance in screening IMP-producing Enterobacteriaceae was evaluated. The system identified 97.8% of IMP producers as being nonsusceptible to imipenem or meropenem, which was higher than that identified by the broth microdilution method (91.3%, imipenem; 41.3%, meropenem).
December 16, 2017: Journal of Microbiological Methods
Pallop Tankaew, Wanwisa Srisawat, Tawatchai Singhla, Khajornsak Tragoolpua, Yasushi Kataoka, Takuo Sawada, Nattawooti Sthitmatee
The ELISA is recognized as an efficient diagnostic tool for antibody detection, but there is no standard ELISA assay for detection of antibodies against hemorrhagic septicemia (HS) in cattle. The present study reports on an indirect ELISA assay for antibody detection of HS in dairy cows, and evaluates the sensitivity (Se) and specificity (Sp) of the method using a Bayesian approach. An indirect ELISA was developed with two types of heat extract antigens, Pasteurella multocida strains P-1256 and M-1404, as coating antigens...
December 12, 2017: Journal of Microbiological Methods
B Horton, F Katzer, M P Y Desmulliez, H L Bridle
Continuous and reliable monitoring of water sources for human consumption is imperative for public health. For protozoa, which cannot be multiplied efficiently in laboratory settings, concentration and recovery steps are key to a successful detection procedure. Recently, the use of megasonic energy was demonstrated to recover Cryptosporidium from commonly used water industry filtration procedures, forming thereby a basis for a simplified and cost effective method of elution of pathogens. In this article, we report the benefits of incorporating megasonic sonication into the current methodologies of Giardia duodenalis elution from an internationally approved filtration and elution system used within the water industry, the Filta-Max®...
December 12, 2017: Journal of Microbiological Methods
Jesus D Castaño, Jiwei Zhang, Jonathan S Schilling
Hydrogen peroxide (H2O2) plays a critical role in generating oxygen radicals as fungi attack and deconstruct plant cell walls. Its concentrations in planta, however, are often low during decomposition and evade detection by traditional methods. Here, we compared relevant methods and selected the best based on detection limits and selectivity.
December 11, 2017: Journal of Microbiological Methods
Joonhong Park, Woori Jang, Myungshin Kim, Yonggoo Kim, So Youn Shin, Kuhn Park, Myung Sook Kim, Soyoung Shin
The increasing burden of multidrug resistant (MDR)-TB, defined by resistance to rifampin (RFP) and isoniazid (INH), and extensively drug resistant-TB, defined by MDR-TB with additional resistance to fluoroquinolones (FQs) and more than one second-line injectable drug, is a serious impediment to global TB control. We evaluated the feasibility of full-length gene analysis including inhA, katG, rpoB, pncA, rpsL embB, eis, and gyrA using a semiconductor NGS with the Ion AmpliSeq TB panel to directly analyse 34 sputum specimens confirmed by phenotypic DST: INH, RFP, ethambutol (EMB), pyrazinamide (PZA), amikacin, kanamycin, streptomycin (SM), FQs including ofloxacin, moxifloxacin, and levofloxacin...
December 11, 2017: Journal of Microbiological Methods
Saman Saadat, Mohammad M Sajadi, Mohammad Yousef Alikhani, Zahra Rikhtegaran Tehrani, Rasoul Yousefi Mashouf
INTRODUCTION: Mycoplasma hominis is an opportunistic pathogen of the human genital tract. Detection of antibodies against this organism in human serum or plasma is theoretically unreliable because of high variation in bacterial surface antigens. In this study, we applied the bioinformatics tools to design a chimeric protein constructed of specific, conserved and predicted immuno-dominant epitopes from two different membrane proteins, P120 and P80. MATERIAL AND METHODS: Linear B-cell epitopes of P120 and P80 were predicted and evaluated by bioinformatics tools and the designed chimeric protein was expressed in Escherichia coli...
December 4, 2017: Journal of Microbiological Methods
Rosanna C Hennessy, Line Christiansen, Stefan Olsson, Peter Stougaard
Fluorescence-based reporter systems are valuable tools for studying gene expression dynamics in living cells. Here we describe a dual-fluorescence reporter system carrying the red fluorescent marker mCherry and the blue fluorescent protein EBFP2 enabling the simultaneous analysis of two promoters in broad-host range autofluorescent Gram-negative bacteria.
December 1, 2017: Journal of Microbiological Methods
Rebecca J Abraham, Mark O'Dea, Bertha Rusdi, Stephen W Page, Ryan O'Handley, Sam Abraham
This study describes a neonatal mouse model of Giardia infection for development of novel antigiardials. Mice were infected with the axenically cultured Assemblage A BAH2c2 strain, with 105 trophozoites per animal recovered. This model proved to be robust and consistent for use in preliminary drug efficacy trials and drug development.
November 30, 2017: Journal of Microbiological Methods
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