scholarly journals Monitoring of Ralstonia eutropha KT1 in Groundwater in an Experimental Bioaugmentation Field by In Situ PCR

2002 ◽  
Vol 68 (1) ◽  
pp. 412-416 ◽  
Author(s):  
Katsuji Tani ◽  
Masahiro Muneta ◽  
Kanji Nakamura ◽  
Katsutoshi Shibuya ◽  
Masao Nasu
Keyword(s):  
Ralstonia Eutropha ◽  
Total Concentration ◽  
Phenol Hydroxylase ◽  
Bacterial Suspension ◽  
Bacterial Cells ◽  
In Situ Pcr ◽  
Hydroxylase Gene ◽  
Pcr Targeting ◽  
Number Of Bacteria

ABSTRACT Ralstonia eutropha KT1, which degrades trichloroethylene, was injected into the aquifer after activation with toluene, and then the number of bacteria was monitored by in situ PCR targeting the phenol hydroxylase gene and by fluorescent in situ hybridization (FISH) targeting 16S rRNA. Before injection of the bacterial suspension, the total concentration of bacteria in the groundwater was approximately 3 × 105 cells/ml and the amount of Ralstonia and bacteria carrying the phenol hydroxylase gene as a percentage of total bacterial cells was less than 0.1%. The concentration of bacteria carrying the phenol hydroxylase gene detected by in situ PCR was approximately 3 × 107 cells/ml 1 h after injection, and the concentration of Ralstonia detected by FISH was similar. The number of bacteria detected by in situ PCR was similar to that detected by FISH 4 days after the start of the extraction of groundwater. On and after day 7, however, the number of bacterial cells detected by FISH was less than that detected by in situ PCR.

scholarly journals Visualization and Direct Counting of Individual Denitrifying Bacterial Cells in Soil by nirK-Targeted Direct in situ PCR

2011 ◽  
Vol 26 (1) ◽  
pp. 74-80 ◽  
Author(s):  
Noriko Ryuda ◽  
Tomoyoshi Hashimoto ◽  
Daisuke Ueno ◽  
Koichi Inoue ◽  
Takashi Someya
Keyword(s):  
Bacterial Cells ◽  
In Situ Pcr ◽  
Direct Counting

Preliminary Study on In Situ Realtime Quantitation of Target Bacteria on the Principle of Flow Cytometry

2017 ◽  
Vol 262 ◽  
pp. 224-227
Author(s):  
Gen Murakami ◽  
Yuichi Sugai ◽  
Kyuro Sasaki
Keyword(s):  
Flow Cytometry ◽  
Bacterial Culture ◽  
Scattered Light ◽  
Bacterial Suspension ◽  
Culture Solution ◽  
Bacterial Cells ◽  
Monitoring Wells ◽  
Measurement Principle ◽  
Preliminary Study

In-situ realtime method that can monitor the target bacteria should be used to determine the real situation of the bacteria in deep parts of heaps in heap bioleaching plants. This study suggest to apply flow cytometry technology to in-situ realtime monitoring of target bacteria. Flow cytometry is a method that can rapidly quantify the bacterial cells in bacterial suspension based on the detection of lights that are emitted from bacterial cells. In this study, we estimated the possibility of the application of flow cytometry to the selective detection of target bacteria. The bacterial culture solution that had been diluted by water including other bacteria was provided for fluorescence spectral analysis and scattered light analysis that were functions of flow cytometry. Our target bacteria could be selectively detected by those analyses in this study, therefore, it was shown that the flow cytometry could be useful for detecting target bacteria selectively. Because the measurement principle of flow cytometry is quite simple, it can be expected to be installed into deep heaps through the monitoring wells and determine the dominance of target bacteria in-situ and realtime in the future.

Development of a Direct In Situ PCR Method for Detection of Specific Bacteria in Natural Environments

1998 ◽  
Vol 64 (4) ◽  
pp. 1536-1540 ◽  
Author(s):  
Katsuji Tani ◽  
Ken Kurokawa ◽  
Masao Nasu
Keyword(s):  
Alkaline Phosphatase ◽  
Single Cell ◽  
Natural Environments ◽  
Single Cell Level ◽  
Bacterial Cells ◽  
Cell Level ◽  
In Situ Pcr ◽  
Glass Slides ◽  
Pcr Products

ABSTRACT We applied HNPP (2-hydroxy-3-naphthoic acid-2′-phenylanilide phosphate) to direct in situ PCR for the routine detection of specific bacterial cells at the single-cell level. PCR was performed on glass slides with digoxigenin-labeled dUTP. The digoxigenin-labeled PCR products were detected with alkaline phosphatase-labeled antidigoxigenin antibody and HNPP which was combined with Fast Red TR. A bright red fluorescent signal was produced from conversion to HNP (dephosphorylated form) by alkaline phosphatase. We used the ECOL DNA primer set for amplification of ribosomal DNA of Escherichia coli to identify cells specifically at the single-cell level in a bacterial mixture. High-contrast images were obtained under an epifluorescence microscope with in situ PCR. By image analysis,E. coli cells in polluted river water also were detected.

scholarly journals Extensive Set of 16S rRNA-Based Probes for Detection of Bacteria in Human Feces

2002 ◽  
Vol 68 (6) ◽  
pp. 2982-2990 ◽  
Author(s):  
Hermie J. M. Harmsen ◽  
Gerwin C. Raangs ◽  
Tao He ◽  
John E. Degener ◽  
Gjalt W. Welling
Keyword(s):  
16S Rrna ◽  
Fluorescent In Situ Hybridization ◽  
Anaerobic Bacteria ◽  
Bacterial Cells ◽  
Human Feces ◽  
Human Gut ◽  
Fecal Samples ◽  
Bacterial Groups ◽  
Number Of Bacteria

ABSTRACT For the detection of six groups of anaerobic bacteria in human feces, we designed seven new 16S rRNA-based oligonucleotide probes. This set of probes extends the current set of probes and gives more data on the composition of the human gut flora. Probes were designed for Phascolarctobacterium and relatives (Phasco741), Veillonella (Veil223), Eubacterium hallii and relatives (Ehal1469), Lachnospira and relatives (Lach571), and Eubacterium cylindroides and relatives (Ecyl387), and two probes were designed for Ruminococcus and relatives (Rbro730 and Rfla729). The hybridization conditions for the new probes were optimized for fluorescent in situ hybridization, and the probes were validated against a set of reference organisms. The probes were applied to fecal samples of 11 volunteers to enumerate their target bacterial groups. The Phasco741 and Veil223 probes both detected average numbers below 1% of the total number of bacteria as determined with the bacterial kingdom-specific Bact338 probe. The Ecyl387 probe detected about 1.4%, the Lach571 and Ehal1469 probes detected 3.8 and 3.6%, respectively, and a combination of the Rbro730 and Rfla729 probes detected 10.3%. A set of 15 probes consisting of probes previously described and those presented here were evaluated in hybridization with the fecal samples of the same volunteers. Together, the group-specific probes detected 90% of the total bacterial cells.

scholarly journals Establishment of novel in vitro culture system with the ability to reproduce oral biofilm formation on dental materials

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Tomoki Kohno ◽  
Haruaki Kitagawa ◽  
Ririko Tsuboi ◽  
Yuma Nishimura ◽  
Satoshi Imazato
Keyword(s):  
Culture System ◽  
Volume Fraction ◽  
Sucrose Solution ◽  
Glass Ionomer Cement ◽  
Bacterial Suspension ◽  
Bacterial Cells ◽  
Oral Biofilm ◽  
Restorative Materials

AbstractIntensive research has been conducted with the aim of developing dental restorative/prosthetic materials with antibacterial and anti-biofilm effects that contribute to controlling bacterial infection in the oral cavity. In situ evaluations were performed to assess the clinical efficacy of these materials by exposing them to oral environments. However, it is difficult to recruit many participants to collect sufficient amount of data for scientific analysis. This study aimed to assemble an original flow-cell type bioreactor equipped with two flow routes and assess its usefulness by evaluating the ability to reproduce in situ oral biofilms formed on restorative materials. A drop of bacterial suspension collected from human saliva and 0.2% sucrose solution was introduced into the assembled bioreactor while maintaining the incubation conditions. The bioreactor was able to mimic the number of bacterial cells, live/dead bacterial volume, and volume fraction of live bacteria in the in situ oral biofilm formed on the surface of restorative materials. The usefulness of the established culture system was further validated by a clear demonstration of the anti-biofilm effects of a glass-ionomer cement incorporating zinc-releasing glasses when evaluated by this system.

Природные материалы для сорбционной очистки хром- и кадмийсодержащих инфильтратов полигонов ТБО

2019 ◽  
pp. 13-19
Author(s):  
A. Safonov ◽  
N. Andriushchenko ◽  
N. Popova ◽  
K. Boldyrev
Keyword(s):  
Sorption Capacity ◽  
Electron Acceptors ◽  
Bacterial Cells ◽  
Maximum Sorption Capacity ◽  
Sorption Material ◽  
Maximum Sorption ◽  
Properties Of Materials ◽  
Solid Waste Landfills ◽  
Sorption Characteristics

Проведен анализ сорбционных характеристик природных материалов (вермикулит, керамзит, перлит, цеолит Трейд ) при очистке кадмий- и хромсодержащих сточных вод с высокой нагрузкой по ХПК. Установлено, что цеолит обладает максимальными сорбционными характеристиками для Cd и Cr и наименьшим биологическим обрастанием. При использовании вермикулита и керамзита или смесей на их основе можно ожидать увеличения сорбционной емкости для Cd и Сr при микробном обрастании, неизбежно происходящем в условиях контакта с водами, загрязненными органическими соединениями и биогенами. При этом биообрастание может повысить иммобилизационную способность материалов для редоксзависимых металлов за счет ферментативных ресурсов бактериальных клеток, использующих их в качестве акцепторов электронов. Эффект микробного обрастания разнонаправленно изменял параметры материалов: для Cr в большинстве случаев уменьшение и для Cd значительное увеличение. При этом дополнительным эффектом иммобилизации Cr является его биологическое восстановление биопленками. Варьируя состав сорбционного материала, можно подбирать смеси, оптимально подходящие для очистки вод инфильтратов с полигонов твердых бытовых отходов с высокой нагрузкой по ХПК и биогенным элементам как при использовании in situ, так и в системах на поверхности.The analysis of the sorption characteristics of natural materials (vermiculite, expanded clay, perlite, Trade zeolite) during the purification of cadmium and chromium-containing leachate with a high COD load was carried out. It was determined that zeolite had the maximum sorption capacity for Cd and Cr and the lowest biological fouling. When using vermiculite and expanded clay or mixtures on their basis, one can expect an increase in the sorption capacity for Cd and Cr during microbial fouling that inevitably occurs during contacting with water polluted with organic compounds and nutrients. In this case biofouling can increase the immobilization properties of materials for redox-dependent metals due to the enzymatic resources of bacterial cells that use them as electron acceptors. The effect of microbial fouling changed the parameters of materials in different directions: for Cr, in most cases, downward, and for Cd, significantly upward. Moreover, chromium biological recovery by biofilms is an additional effect of immobilization. Varying the composition of the sorption material provides for selecting mixtures that are optimally suitable for the purification of leachates from solid waste landfills with high COD and nutrients load, both when used in situ and in surface systems.

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