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Live/dead baclight kit

Laura Purevdorj-Gage, Brian Nixon, Kyle Bodine, Qilong Xu, William T Doerrler
A method for microscopic enumeration of viable Salmonella enterica in meat samples was developed by using the LIVE/DEAD BacLight kit technology. A two-step centrifugation and wash process was developed to clean the samples from food and chemical impurities that might otherwise interfere with the appropriate staining reactions. The accuracy of the BacLight kit-based viability assessments was confirmed with various validation tests that were conducted by following the manufacturer's instructions. For the biocide challenge tests, chicken parts each bearing around 8...
March 2018: Journal of Food Protection
Laia Fernández-Barat, Ana Motos, Otavio T Ranzani, Gianluigi Li Bassi, Elisabet Aguilera Xiol, Tarek Senussi, Chiara Travierso, Chiara Chiurazzi, Francesco Idone, Laura Muñoz, Jordi Vila, Miquel Ferrer, Paolo Pelosi, Francesco Blasi, Massimo Antonelli, Antoni Torres
Microorganisms are able to form biofilms within respiratory secretions. Methods to disaggregate such biofilms before utilizing standard, rapid, or high throughput diagnostic technologies may aid in pathogen detection during ventilator associated pneumonia (VAP) diagnosis. Our aim was to determine if sonication of endotracheal aspirates (ETA) would increase the sensitivity of qualitative, semi-quantitative, and quantitative bacterial cultures in an animal model of pneumonia caused by Pseudomonasaeruginosa or by methicillin resistant Staphylococcusaureus (MRSA)...
September 20, 2017: Microorganisms
Sopita Ruangcharoen, Waraporn Suwannarong, Marie Rossini Carmela T Lachica, Jan G M Bolscher, Kamran Nazmi, Watcharee Khunkitti, Suwimol Taweechaisupapong
Lactoferrin chimera (LFchimera), a heterodimeric peptide containing lactoferrampin (LFampin265-284) and a part of lactoferricin (LFcin17-30), possesses a broad spectrum of antimicrobial activity. However, there is no report on the inhibitory effects of LFchimera against multispecies oral biofilms. This study aimed to determine the effects of LFchimera in comparison to chlorhexidine digluconate (CHX) and minocycline hydrochloride (MH), on in vitro multispecies biofilms derived from subgingival plaque of periodontitis patients harboring Aggregatibacter actinomycetemcomitans...
August 19, 2017: World Journal of Microbiology & Biotechnology
Junyan Liu, Lin Li, Bing Li, Brian M Peters, Yang Deng, Zhenbo Xu, Mark E Shirtliff
OBJECTIVE: The present study aimed at investigating the capability of L. plantarum strain BM-LP14723 to enter into and recover from the viable but nonculturable (VBNC) state and to cause beer spoilage. METHODS: VBNC state was induced by incubating in beer with subculturing or low temperature treatment. Culturable, total, and viable cells numbers were assessed by MRS agar plate counting, acridine orange direct counting, and Live/Dead BacLight bacterial viability kit, respectively...
September 2017: Microbial Pathogenesis
Junyan Liu, Lin Li, Bing Li, Brian M Peters, Yang Deng, Zhenbo Xu, Mark E Shirtliff
OBJECTIVES: This study aimed to investigate the spoilage capability of Lactobacillus lindneri during the induction and resuscitation of viable but nonculturable (VBNC) state. METHODS: L. lindneri strain was identified by sequencing the PCR product (amplifying 16S rRNA gene) using ABI Prism 377 DNA Sequencer. During the VBNC state induction by low temperature storage and beer adaption, total, culturable, and viable cells were assessed by acridine orange direct counting, plate counting, and Live/Dead BacLight bacterial viability kit, respectively...
June 2017: Microbial Pathogenesis
Igor Tsesis, Shlomo Elbahary, Nuphar Blau Venezia, Eyal Rosen
OBJECTIVES: The purpose of this study was to evaluate Enterococcus faecalis colonization at the apical part of root canals following root-end resection and filling using confocal laser scanning microscopy (CLSM). MATERIALS AND METHODS: The apical 3-mm root-ends of 55 extracted single rooted human teeth were resected, and 3-mm retrograde cavities were prepared and filled using either mineral trioxide aggregate (MTA), intermediate restorative material (IRM), or Biodentine (n = 10 each); 25 teeth served as controls...
January 2018: Clinical Oral Investigations
Wei Hu, Kotaro Murata, Daizhou Zhang
Rainwater contains substantial bacteria and rain is an efficient pathway for the dissemination of bacteria from the atmosphere to land and water surfaces. However, quantitative information on rainwater bacteria is very limited due to the lack of a reliable method. In this study, the epifluorescence microscopy enumeration with the LIVE/DEAD BacLight Bacterial Viability Kit stain was verified to quantify the abundance of viable and non-viable bacterial cells in rainwater, with the 4',6-diamidino-2-phenylindole (DAPI) stain for the reference of total cell counts...
January 2017: Journal of Environmental Sciences (China)
Rafaela Pignatti de Freitas, Vanessa Raquel Greatti, Murilo Priori Alcalde, Bruno Cavalini Cavenago, Rodrigo Ricci Vivan, Marco Antonio Hungaro Duarte, Ana Carolina Villas Bôas Weckwerth, Paulo Henrique Weckwerth
INTRODUCTION: The objective of the present study was to evaluate the in vitro antibiofilm activity and pH of calcium hydroxide associated with different nonsteroidal anti-inflammatory drugs (NSAIDs). METHODS: The groups analyzed were as follows: group 1, calcium hydroxide paste with propylene glycol; group 2, calcium hydroxide paste with propylene glycol + 5% diclofenac sodium; group 3, calcium hydroxide paste with propylene glycol + 5% ibuprofen; group 4, calcium hydroxide paste with propylene glycol + 5% ciprofloxacin; and group 6, positive control (without medication)...
January 2017: Journal of Endodontics
Wei Qiu, Biao Ren, Huanqin Dai, Lixin Zhang, Qiong Zhang, Xuedong Zhou, Yuqing Li
OBJECTIVE: The aim of this study was to determine the inhibitory effect of eight antifungal drugs on S. mutans growth, biofilm formation and virulence factors. METHODS: The actions of antifungal drugs on S. mutans were determined by recovery plates and survival kinetic curves. Biofilms were observed by scanning electron microscopy and the viable cells were recovered on BHI plates, meanwhile biofilms were stained by BacLight live/dead kit to investigate the biofilm viability...
January 2017: Archives of Oral Biology
Eun-Hee Kim, Sun-Young Kang, Bog-Im Park, Young-Hoi Kim, Young-Rae Lee, Jin-Hee Hoe, Na-Young Choi, Ji-Young Ra, So-Youn An, Yong-Ouk You
Chamaecyparis obtusa (C. obtusa) is known to have antimicrobial effects and has been used as a medicinal plant and in forest bathing. This study aimed to evaluate the anticariogenic activity of essential oil of C. obtusa on Streptococcus mutans, which is one of the most important bacterial causes of dental caries and dental biofilm formation. Essential oil from C. obtusa was extracted, and its effect on bacterial growth, acid production, and biofilm formation was evaluated. C. obtusa essential oil exhibited concentration-dependent inhibition of bacterial growth over 0...
2016: Evidence-based Complementary and Alternative Medicine: ECAM
Ying-ting Yang, Yan Wei, Bin Xia, Zhen-gao Wang
OBJECTIVE: To study the effects of cells'and bacteria's adhesion and proliferation on different fiber diameters of polypyrrole coating with electricity. METHODS: Titanium coated with polypyrrole was divided into no electrical stimulation and stimulation groups, each group had 30-60 nm, 70- 100 nm, 130-170 nm diameters of the fiber. MC3T3 cells and Staphylococcus aureus (S. aureus) were inoculated on different fiber diameters of polypyrrole coating with and without electric stimulation...
February 18, 2016: Beijing da Xue Xue Bao. Yi Xue Ban, Journal of Peking University. Health Sciences
Janine Heise, Marcella Nega, Mashal Alawi, Dirk Wagner
In clinical trials investigating human health and in the analysis of microbial communities in cultures and natural environments, it is a substantial challenge to differentiate between living, potentially active communities and dead cells. The DNA-intercalating dye propidium monoazide (PMA) enables the selective masking of DNA from dead, membrane-compromised cells immediately before DNA extraction. In the present study, we evaluated for the first time a PMA treatment for methanogenic archaea in cultures and particle-rich environmental samples...
February 2016: Journal of Microbiological Methods
Diana Marcela Castillo, Yormaris Castillo, Nathaly Andrea Delgadillo, Yineth Neuta, Johana Jola, Justo Leonardo Calderón, Gloria Inés Lafaurie
This study investigated the effect of hypochlorous acid (HOCl) rinses and chlorhexidine (CHX) on the bacterial viability of S. mutans, A. israelii, P. gingivalis, A. actinomycetemcomitans, E. corrodens, C. rectus, K. oxytoca, K. pneumoniae and E. cloacae. The percentage of live bacteria was tested by fluorescence method using Live/Dead kit(r) and BacLight (Molecular Probes(r)) and compared between groups by the Kruskal-Wallis and U Mann-Whitney tests with Bonferroni correction (p value<0.012). The effect of HOCl and CHX on total proteins of P...
October 2015: Brazilian Dental Journal
D R Yarullina, L G Damshkaln, N L Bruslik, O A Konovalova, O N Ilinskaya, V I Lozinsky
BACKGROUND: Probiotics are live microorganisms, generally either lactobacilli or bifidobacteria, which when administered in adequate amounts confer a health benefit to the host [1]. Due to the growing evidence of health benefits associated with their use, probiotics are of increasing interest and represent now a significant growth area in the functional foods industry [2]. However, to be effective, orally administered probiotics should survive preparation of dosage forms and passage through acidic environment of the gastrointestinal tract (GIT)...
2015: International Journal of Risk & Safety in Medicine
C Shi, Z Jia, Y Chen, M Yang, X Liu, Y Sun, Z Zheng, X Zhang, K Song, L Cui, A B Baloch, X Xia
AIMS: The objective of this study was to determine the combined effect of thymoquinone (TQ) and mild heat on Cronobacter sakazakii in reconstituted infant formula. METHODS AND RESULTS: Reconstituted infant formula samples inoculated with a mixture of four C. sakazakii strains (approx. 6·5 log CFU ml(-1) ) were prepared with various concentrations of TQ (0, 5, 10, 20 and 30 mmol l(-1) ) and were heated to 45, 50 and 55°C for 0, 10, 20, 30, 60 and 120 min, and the surviving populations of C...
December 2015: Journal of Applied Microbiology
Karlo Marinic, Daniel Manoil, Anna Filieri, John C Wataha, Jacques Schrenzel, Norbert Lange, Serge Bouillaguet
BACKGROUND: In dentistry, antibacterial photodynamic therapy (a-PDT) has shown promising results for inactivating bacterial biofilms causing carious, endodontic and periodontal diseases. In the current study, we assessed the ability of eosin Y exposed to 3 irradiation protocols at inactivating Enterococcus faecalis biofilms, in vitro. METHODS: E. faecalis biofilms formed on hydroxyapatite disks were incubated with eosin Y (10-80μM), then activated with blue light using different irradiation protocols...
September 2015: Photodiagnosis and Photodynamic Therapy
Nicole Zacharias, Thomas Kistemann, Christiane Schreiber
The paper compares two methods of distinguishing between alive and dead cells by differentiation on the basis of their membrane structure: LIVE/DEAD flow cytometry and PMA-qPCR. LIVE/DEAD flow cytometry was established using the LIVE/DEAD(®) BacLight™ Bacterial Viability Kit with different ratios of Legionella pneumophila and Escherichia coli cells with intact and compromised membranes (heat treated). The PMA-qPCR method was tested and modified, and results were compared with those from LIVE/DEAD flow cytometry using L...
November 2015: International Journal of Hygiene and Environmental Health
Marc Philipp Dittmer, Carolina Fuchslocher Hellemann, Sebastian Grade, Wieland Heuer, Meike Stiesch, Rainer Schwestka-Polly, Anton Phillip Demling
INTRODUCTION: The purpose of the present study was to investigate and compare early biofilm formation on biomaterials, which are being used in contemporary fixed orthodontic treatment. METHODS: This study comprised 10 healthy volunteers (5 females and 5 males) with a mean age of 27.3 +-3.7 years. Three slabs of different orthodontic materials (stainless steel, gold and ceramic) were placed in randomized order on a splint in the mandibular molar region. Splints were inserted intraorally for 48 h...
2015: Head & Face Medicine
Tylor J Johnson, Michael B Hildreth, Liping Gu, Ruanbao Zhou, William R Gibbons
Filamentous cyanobacteria are currently being engineered to produce long-chain organic compounds, including 3rd generation biofuels. Because of their filamentous morphology, standard methods to quantify viability (e.g., plate counts) are not possible. This study investigated a dual-fluorescence assay based upon the LIVE/DEAD® BacLight™ Bacterial Viability Kit to quantify the percent viability of filamentous cyanobacteria using a microplate reader in a high throughput 96-well plate format. The manufacturer's protocol calls for an optical density normalization step to equalize the numbers of viable and non-viable cells used to generate calibration curves...
June 2015: Journal of Microbiological Methods
Paola Sepulveda-Medina, Yelena Katsenovich, Vishal Musaramthota, Michelle Lee, Brady Lee, Rupak Dua, Leonel Lagos
Past disposal practices at nuclear production facilities have led to the release of liquid waste into the environment creating multiple radionuclide plumes. Microorganisms are known for the ability to interact with radionuclides and impact their mobility in soils and sediments. Gram-positive Arthrobacter sp. are one of the most common bacterial groups in soils and are found in large numbers in subsurface environments contaminated with radionuclides. This study experimentally analyzed changes on the bacteria surface at the nanoscale level after uranium exposure and evaluated the effect of aqueous bicarbonate ions on U(VI) toxicity of a low uranium-tolerant Arthrobacter oxydans strain G968 by investigating changes in adhesion forces and cell dimensions via atomic force microscopy (AFM)...
June 2015: Research in Microbiology
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