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Journal of Microbiological Methods

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https://www.readbyqxmd.com/read/28411150/comparative-diagnostic-efficacy-of-recombinant-llo-and-pi-plc-based-elisas-for-detection-of-listeriosis-in-animals
#1
Rahul D Suryawanshi, Satya Veer Singh Malik, Bhushan Jayarao, Sandeep P Chaudhari, Emily Savage, Jess Vergis, Nitin V Kurkure, Sukhadeo B Barbuddhe, Deepak B Rawool
The present study for the first time evaluates the serodiagnostic efficacy of two recombinant antigens namely, listeriolysin O (rLLO) and phosphatidyl-inositol phospholipase C (rPI-PLC). Indirect ELISA with the above recombinant antigens was used on samples collected from bovines (n=106), goats (n=138) and pigs (n=92) having either a history of abortion, emaciation and/or apparently healthy animals. Isolation of Listeria was attempted from the blood samples using USDA-FSIS method. On screening of test sera by rLLO-based ELISA, antibodies against anti-listeriolysin O (ALLO) were observed in goats (22...
April 11, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28404228/dynamic-light-scattering-a-fast-and-reliable-method-to-analyze-bacterial-growth-during-the-lag-phase
#2
Susana Vargas, Blanca E Millán-Chiu, Sofía M Arvizu-Medrano, Achim M Loske, Rogelio Rodríguez
A comparison between plate counting (PC) and dynamic light scattering (DLS) is reported. PC is the standard technique to determine bacterial population as a function of time; however, this method has drawbacks, such as the cumbersome preparation and handling of samples, as well as the long time required to obtain results. Alternative methods based on optical density are faster, but do not distinguish viable from non-viable cells. These inconveniences are overcome by using DLS. Two different bacteria strains were considered: Escherichia coli and Staphylococcus aureus...
April 9, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28396241/molecular-typing-of-legionella-pneumophila-from-air-conditioning-cooling-waters-using-mip-gene-sbt-and-faflp-methods
#3
Xiangli Gong, Juntao Li, Ying Zhang, Shuiping Hou, Pinghua Qu, Zhicong Yang, Shouyi Chen
Legionella spp. are important waterborne pathogens. Molecular typing has become an important method for outbreaks investigations and source tracking of Legionnaires. In a survey program conducted by the Guangzhou Center for Disease Control and Prevention, multiple serotypes Legionella pneumophila (L. pneumophila) were isolated from waters in air-conditioning cooling towers in urban Guangzhou region, China between 2008 and 2011. Three genotyping methods, mip (macrophage infectivity potentiator) genotyping, SBT (sequence-based typing), and FAFLP (fluorescent amplified fragment length polymorphism analysis) were used to type these waterborne L...
April 7, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28392069/corrigendum-to-a-spectrophotometric-screening-method-for-avermectin-oxidizing-microorganisms-j-microbiol-methods-135-2017-93-95
#4
Yuan-Shan Wang, Qi-Wei Hu, Xing-Chang Zheng, Jian-Fen Zhang, Yu-Guo Zheng
No abstract text is available yet for this article.
April 6, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28390706/comparison-of-imipenem-and-meropenem-antibiotics-for-the-maldi-tof-ms-detection-of-carbapenemase-activity
#5
Veronika Rotova, Costas C Papagiannitsis, Anna Skalova, Katerina Chudejova, Jaroslav Hrabak
A comparison of carbapenem molecules for the detection of carbapenemase-producing bacteria by MALDI-TOF MS showed that imipenem exhibited higher sensitivity (97%) and specificity (100%) scores for Pseudomonas aeruginosa than meropenem. However, meropenem was more efficient (98% sensitivity and 100% specificity) against Enterobacteriaceae.
April 5, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28385456/real-time-pcr-assay-for-aquimarina-macrocephali-subsp-homaria-and-its-distribution-in-shell-disease-lesions-of-homarus-americanus-milne-edwards-1837-and-environmental-samples
#6
Robert A Quinn, Suchandra Hazra, Roxanna Smolowitz, Andrei Y Chistoserdov
Epizootic shell disease (ESD) is causing major losses to the lobster fishery in southern New England. Potential pathogens have been identified in lesion communities, but there are currently no efficient means of detecting and quantifying their presence. A qPCR assay was developed for a key potential pathogen, Aquimarina macrocephali subsp. homaria found to be ubiquitous in ESD lesions but not the unaffected integument. Application of the assay to various samples demonstrated that A. macrocephali subsp. homaria is ubiquitous and abundant in lobster lesions, commonly associated with healthy surfaces of crabs and is scarce in water and sediment samples from southern New England suggesting the affinity of this microorganism to the Arthropod integument...
April 3, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28359738/development-of-a-multiplex-real-time-pcr-assay-for-phylogenetic-analysis-of-uropathogenic-escherichia-coli
#7
Mojtaba Hasanpour, Akram Najafi
Uropathogenic Escherichia coli (UPEC) is among major pathogens causing 80-90% of all episodes of urinary tract infections (UTIs). Recently, E. coli strains are divided into eight main phylogenetic groups including A, B1, B2, C, D, E, F, and clade I. This study was aimed to develop a rapid, sensitive, and specific multiplex real time PCR method capable of detecting phylogenetic groups of E. coli strains. This study was carried out on E. coli strains (isolated from the patient with UTI) in which the presence of all seven target genes had been confirmed in our previous phylogenetic study...
March 28, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28359737/a-flow-cytometric-method-to-evaluate-eosinophil-mediated-uptake-of-probiotic-lactobacillus-reuteri
#8
Laura S Kraemer, Todd A Brenner, Julia O Krumholz, Helene F Rosenberg
Eosinophils are resident leukocytes of gut mucosa. Here we present a combined flow cytometric-antibiotic protection assay to identify mouse eosinophils capable of bacterial uptake, specifically, Gram-positive Lactobacillus reuteri, in studies performed ex vivo. The assay may be adapted for use in vivo.
March 27, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28347725/development-of-novel-monoclonal-antibodies-directed-against-catechins-for-investigation-of-antibacterial-mechanism-of-catechins
#9
Takahisa Miyamoto, Xiaoguang Zhang, Yuuki Ueyama, Kitichalermkiat Apisada, Motokazu Nakayama, Yasuto Suzuki, Tadahiro Ozawa, Asako Mitani, Naofumi Shigemune, Kanami Shimatani, Koji Yui, Ken-Ichi Honjoh
Catechins are major polyphenolic compounds of green tea. To investigate mechanism for antibacterial action of catechins, 11 monoclonal antibodies (MAbs) were raised against a 3-succinyl-epicatechin (EC)-keyhole limpet hemocyanin (KLH) conjugate. Amino acid sequences of variable regions determined for MAbs b-1058, b-1565, and b-2106 confirmed their innovative character. MAb b-1058 strongly interacted with its target substances in the following order of magnitude: theaflavin-3,3'-di-O-gallate (TFDG)>theaflavin-3-O-gallate (TF3G)≥theaflavin-3'-O-gallate (TF3'G)>gallocatechin gallate (GCg)>penta-O-galloyl-β-d-glucose (PGG)>epigallocatechin gallate (EGCg), as determined using surface plasmon resonance (SPR) on MAb-immobilized sensor chips...
March 24, 2017: Journal of Microbiological Methods
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
#10
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
#11
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
#12
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
#13
Marta García-Coca, Ignacio Gadea, Jaime Esteban
No abstract text is available yet for this article.
March 19, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28323067/evaluation-of-a-real-time-impedance-analysis-platform-on-fungal-infection
#14
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
#15
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
#16
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
#17
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
#18
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
#19
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
#20
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
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