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Journal of Microbiological Methods

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https://www.readbyqxmd.com/read/28344084/a-rapid-procedure-for-the-in-situ-assay-of-periplasmic-pqq-dependent-methanol-dehydrogenase-in-intact-single-bacterial-colonies
#1
Venkata Ramana Vemuluri, Shreya Shaw, Caroline Autenrieth, Robin Ghosh
Mechanistic details of methanol oxidation catalyzed by the periplasmically-located pyrroloquinoline quinone-dependent methanol dehydrogenase of methylotrophs can be elucidated using site-directed mutants. Here, we present an in situ colony assay of methanol dehydrogenase, which allows robotic screening of large populations of intact small colonies, and regrowth of colonies for subsequent analysis.
March 23, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28342745/culture-of-non-typeable-haemophilus-influenzae-from-the-nasopharynx-not-all-media-are-equal
#2
Tegan M Harris, Angela Rumaseb, Jemima Beissbarth, Federica Barzi, Amanda J Leach, Heidi C Smith-Vaughan
The efficacy of chocolate agar, versus bacitracin, vancomycin, clindamycin, chocolate agar (BVCCA) for the isolation of non-typeable Haemophilus influenzae (NTHi) from nasopharyngeal swabs was determined. BVCCA cultured NTHi from 97.3% of NTHi-positive swabs, compared to 87.1% for chocolate agar. To maximise culture sensitivity, the use of both media is recommended.
March 22, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28336461/chloroform-free-permeabilization-for-improved-detection-of-%C3%AE-galactosidase-activity-in-vibrio-cholerae
#3
Charlotte Toulouse, Claudia C Häse, Julia Steuber
LacZ (β-galactosidase) is used to monitor the transcription of genes in reporter strains carrying the lacZ gene under the control of a promotor of interest. This protocol for LacZ activity determinations in Vibrio cholerae following detergent lysis results in 2.5-fold increase of LacZ activities compared to lysis with chloroform.
March 21, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28330780/relationship-between-conventional-culture-and-flow-cytometry-for-the-diagnosis-of-urinary-tract-infection
#4
Marta García-Coca, Ignacio Gadea, Jaime Esteban
No abstract text is available yet for this article.
March 18, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28323067/evaluation-of-a-real-time-impedance-analysis-platform-on-fungal-infection
#5
Jiufeng Sun, Dan Ning, Wenying Cai, Huiqiong Zhou, Huan Zhang, Dawei Guan, De Wu
End-point assays of in vitro cell proliferation and death have been employed to study the mechanisms of fungal pathogenesis and have shown the responses of host cells at individual time points. A new cell analysis technology has been developed that allows for the continuous measurement and quantification of cell activities, thus enabling the dynamic assessment of electrical impedance when various pathogens are cultured in vitro. In this study, this system was evaluated to determine the response of the cell line RAW264...
March 18, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28323066/development-of-an-economical-autonomous-phstat-system-for-culturing-phytoplankton-under-steady-state-or-dynamic-conditions
#6
Rachel L Golda, Mark D Golda, Jacqueline A Hayes, Tawnya D Peterson, Joseph A Needoba
Laboratory investigations of physiological processes in phytoplankton require precise control of experimental conditions. Chemostats customized to control and maintain stable pH levels (pHstats) are ideally suited for investigations of the effects of pH on phytoplankton physiology, for example in context of ocean acidification. Here we designed and constructed a simple, flexible pHstat system and demonstrated its operational capabilities under laboratory culture conditions. In particular, the system is useful for simulating natural cyclic pH variability within aquatic ecosystems, such as diel fluctuations that result from metabolic activity or tidal mixing in estuaries...
March 18, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28323065/the-impact-of-fecal-sample-processing-on-prevalence-estimates-for-antibiotic-resistant-escherichia-coli
#7
Sylvia Omulo, Eric T Lofgren, Maina Mugoh, Moshe Alando, Joshua Obiya, Korir Kipyegon, Gilbert Kikwai, Wilson Gumbi, Samuel Kariuki, Douglas R Call
Investigators often rely on studies of Escherichia coli to characterize the burden of antibiotic resistance in a clinical or community setting. To determine if prevalence estimates for antibiotic resistance are sensitive to sample handling and interpretive criteria, we collected presumptive E. coli isolates (24 or 95 per stool sample) from a community in an urban informal settlement in Kenya. Isolates were tested for susceptibility to nine antibiotics using agar breakpoint assays and results were analyzed using generalized linear mixed models...
March 18, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28285169/microbial-profiling-of-cpn60-universal-target-sequences-in-artificial-mixtures-of-vaginal-bacteria-sampled-by-nylon-swabs-or-self-sampling-devices-under-different-storage-conditions
#8
John J Schellenberg, Angela Yena Oh, Janet E Hill
The vaginal microbiome is increasingly characterized by deep sequencing of universal genes. However, there are relatively few studies of how different specimen collection and sample storage and processing influence these molecular profiles. Here, we evaluate molecular microbial community profiles of samples collected using the HerSwab™ self-sampling device, compared to nylon swabs and under different storage conditions. In order to minimize technical variation, mixtures of 11 common vaginal bacteria in simulated vaginal fluid medium were sampled and DNA extracts prepared for massively parallel sequencing of the cpn60 universal target (UT)...
March 9, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28285168/design-and-use-of-mouse-control-dna-for-dna-biomarker-extraction-and-pcr-detection-from-urine-application-for-transrenal-mycobacterium-tuberculosis-dna-detection
#9
Hali Bordelon, Keersten M Ricks, Megan E Pask, Patricia K Russ, Francesca Solinas, Mark L Baglia, Philip A Short, Andrew Nel, Jonathan Blackburn, Keertan Dheda, Carlos Zamudio, Tatiana Cáceres, David W Wright, Frederick R Haselton, April C Pettit
Urine samples are increasingly used for diagnosing infections including Escherichia coli, Ebola virus, and Zika virus. However, extraction and concentration of nucleic acid biomarkers from urine is necessary for many molecular detection strategies such as polymerase chain reaction (PCR). Since urine samples typically have large volumes with dilute biomarker concentrations making them prone to false negatives, another impediment for urine-based diagnostics is the establishment of appropriate controls particularly to rule out false negatives...
March 9, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28285864/e-coli-reca-gene-improves-gene-targeted-homologous-recombination-in-mycoplasma-hyorhinis
#10
Hassan Z A Ishag, Qiyan Xiong, Maojun Liu, Zhixin Feng, Guoqing Shao
Mycoplasma hyorhinis is an opportunistic pathogen of pigs. Recently, it has been shown to transform cell cultures, increasing the attention of the researchers. Studies on the pathogenesis require specific genetic tool that is not yet available for the pathogen. To address this limitation, we constructed two suicide plasmids pGEMT-tetM/LR and pGEMT-recA-tetM/LR having a tetracycline resistance marker flanked by two hemolysin gene arms. The latter plasmid encodes an E. coli recA, a gene involved in DNA recombination, repair and maintenance of DNA...
March 8, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28285167/the-investigation-of-the-truncated-mbta-gene-within-the-mycobactin-cluster-of-mycobacterium-avium-subspecies-paratuberculosis-as-a-novel-diagnostic-marker-for-real-time-pcr
#11
Marcel de Kruijf, Aidan Coffey, Jim O'Mahony
The inability of Mycobacterium avium subspecies paratuberculosis (MAP) to produce endogenous mycobactin in-vitro is most likely due to the presence of a truncated mbtA gene within the mycobactin cluster of MAP. The main goal of this study was to investigate this unique mbtA truncation as a potential novel PCR diagnostic marker for MAP. Novel primers were designed that were located within the truncated region and the contiguous MAP2179 gene. Primers were evaluated against non-MAP isolates and no amplicons were generated...
March 8, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28268111/characterization-of-growth-inhibition-of-oral-bacteria-by-sophorolipid-using-a-microplate-format-assay
#12
Daniel K Y Solaiman, Richard D Ashby, Joseph Uknalis
Sophorolipid (SL) is a class of glycolipid biosurfactant produced by yeast and has potent antimicrobial activity against many microorganisms. In this paper, a microplate-based method was developed to characterize the growth inhibition by SL on five representative species of caries-causing oral bacteria. Bacterial growth on microplate in the absence and presence of varying concentrations of SL was continuously monitored by recording the absorbance at 600nm of the cultures using a microplate reader. The results showed that SL completely inhibited the growth of the Lactobacilli at ≥1mg/ml and the Streptococci at much lower concentrations of ≥50μg/ml...
March 6, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28267571/rapid-typing-of-mannheimia-haemolytica-major-genotypes-1-and-2-using-maldi-tof-mass-spectrometry
#13
John Dustin Loy, Michael L Clawson
Genotype 2M. haemolytica predominantly associate over genotype 1 with the lungs of cattle with respiratory disease and ICEs containing antimicrobial resistance genes. Distinct protein masses were detected by MALDI-TOF MS between genotype 1 and 2 strains. MALDI-TOF MS could rapidly differentiate genotype 2 strains in veterinary diagnostic laboratories.
March 4, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28267570/identification-of-microorganisms-grown-on-chromogenic-media-by-maldi-tof-ms
#14
Petra Lüthje, Arthur B Pranada, Duncan Carruthers-Lay, Marc Desjardins, Olivier Gaillot, David Wareham, Holly Ciesielczuk, Volkan Özenci
Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and chromogenic media are widely used in clinical microbiology laboratories to facilitate the rapid selection and identification of pathogens. The aim of this study was to evaluate whether usage of chromogenic media limits the diagnostic performance of MALDI-TOF MS for microbial identification. A total of 386 microorganisms collected and analyzed at five laboratories were included. Isolates were cultured on relevant chromogenic media and non-selective agar plates in parallel and identified using the Bruker MALDI-TOF MS...
March 4, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28259789/evaluation-of-a-serological-test-for-the-diagnosis-of-borrelia-miyamotoi-disease-in-europe
#15
Setareh Jahfari, Denis S Sarksyan, Nadezda M Kolyasnikova, Joppe W Hovius, Hein Sprong, Alexander E Platonov
BACKGROUND: Borrelia miyamotoi causes systemic febrile illness and is transmitted by the same tick species that transmits Borrelia burgdorferi sensu lato and tick-borne encephalitis virus. We describe a serological test using a fragment of glycerophosphodiester phosphodiesterase (GlpQ) as an antigen, and determined its performance in well-defined patient categories. METHODS: Serum of patients with PCR-confirmed Borrelia miyamotoi disease (BMD), Lyme borreliosis (LB), tick-borne encephalitis (TBE), and healthy blood donors (HBD) were collected in Udmurt Republic, Russia...
March 1, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28249794/identification-of-mycobacterial-gara-as-a-substrate-of-protein-kinase-g-from-m-tuberculosis-using-a-kestrel-based-proteome-wide-approach
#16
Philipp Mueller, Jean Pieters
Signal transduction in bacteria is generally mediated via two-component systems. These systems depend on the transfer of a phosphate molecule from a donor to an acceptor by histidine kinases, thereby activating the acceptor to allow downstream signaling/activation. Several bacterial genomes, including the genome of M. tuberculosis, were shown to encode eukaryotic-like kinases. To better understand the function of these kinases and the regulatory networks within which they operate, identification of downstream targets is essential...
February 27, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28238755/application-of-a-stir-bar-sorptive-extraction-sample-preparation-method-with-hplc-for-soil-fungal-biomass-determination-in-soils-from-a-detrital-manipulation-study
#17
Áron Beni, Kate Lajtha, János Kozma, István Fekete
Ergosterol is a sterol found ubiquitously in cell membranes of filamentous fungi. Although concentrations in different fungal species span the range of 2.6 to 42μg/mL of dry mass, many studies have shown a strong correlation between soil ergosterol content and fungal biomass. The analysis of ergosterol in soil therefore could be an effective tool for monitoring changes in fungal biomass under different environmental conditions. Stir Bar Sorptive Extraction (SBSE) is a new sample preparation method to extract and concentrate organic analytes from liquid samples...
February 24, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28235560/proteome-mining-for-the-identification-and-in-silico-characterization-of-putative-drug-targets-of-multi-drug-resistant-clostridium-difficile-strain-630
#18
Mohtashim Lohani, Anupam Dhasmana, Shafiul Haque, Mohd Wahid, Arshad Jawed, Sajad A Dar, Raju K Mandal, Mohammed Y Areeshi, Saif Khan
Clostridium difficile is an enteric pathogen that causes approximately 20% to 30% of antibiotic-associated diarrhea. In recent years, there has been a substantial rise in the rate of C. difficile infections as well as the emergence of virulent and antibiotic resistant C. difficile strains. So, there is an urgent need for the identification of therapeutic potential targets and development of new drugs for the treatment and prevention of C. difficile infections. In the current study, we used a hybrid approach by combining sequence similarity-based approach and protein-protein interaction network topology-based approach to identify and characterize the potential drug targets of C...
February 22, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28232090/immobilization-of-microalgae-cells-in-alginate-facilitates-isolation-of-dna-and-rna
#19
Blanca R Lopez, Juan-Pablo Hernandez, Yoav Bashan, Luz E de Bashan
Isolation of nucleic acids from Chlorella is difficult, given the chemically complex nature of their cell walls and variable production of metabolites. Immobilization of microalgae in polymers adds additional difficulty. Here, we modified, amended, and standardized methods for isolation of nucleic acids and compared the yield of DNA and RNA from free-living and encapsulated microalgae C. sorokiniana. Isolation of nucleic acids from immobilized cells required two steps in dissolving the alginate matrix, releasing the cells, and mechanical disruption with glass beads...
February 20, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28215962/two-colour-fluorescence-fluorimetric-analysis-for-direct-quantification-of-bacteria-and-its-application-in-monitoring-bacterial-growth-in-cellulose-degradation-systems
#20
Kwabena O Duedu, Christopher E French
Monitoring bacterial growth is an important technique required for many applications such as testing bacteria against compounds (e.g. drugs), evaluating bacterial composition in the environment (e.g. sewage and wastewater or food suspensions) and testing engineered bacteria for various functions (e.g. cellulose degradation). T?=1,^FigItem(1) ^ReloadFigure=Yesraditionally, rapid estimation of bacterial growth is performed using spectrophotometric measurement at 600nm (OD600) but this estimation does not differentiate live and dead cells or other debris...
February 17, 2017: Journal of Microbiological Methods
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