Microscopic detection of the toluene dioxygenase gene and its expression inside bacterial cells in seawater using prokaryotic in situ PCR

Author(s):  
F. Chen ◽  
W. A. Dustman ◽  
R. E. Hodson
2011 ◽  
Vol 26 (1) ◽  
pp. 74-80 ◽  
Author(s):  
Noriko Ryuda ◽  
Tomoyoshi Hashimoto ◽  
Daisuke Ueno ◽  
Koichi Inoue ◽  
Takashi Someya

1998 ◽  
Vol 64 (4) ◽  
pp. 1536-1540 ◽  
Author(s):  
Katsuji Tani ◽  
Ken Kurokawa ◽  
Masao Nasu

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.


2002 ◽  
Vol 68 (1) ◽  
pp. 412-416 ◽  
Author(s):  
Katsuji Tani ◽  
Masahiro Muneta ◽  
Kanji Nakamura ◽  
Katsutoshi Shibuya ◽  
Masao Nasu

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.


Author(s):  
A. Safonov ◽  
N. Andriushchenko ◽  
N. Popova ◽  
K. Boldyrev

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.


2017 ◽  
Vol 262 ◽  
pp. 224-227
Author(s):  
Gen Murakami ◽  
Yuichi Sugai ◽  
Kyuro Sasaki

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.


2016 ◽  
Vol 88 (5) ◽  
pp. 2792-2798 ◽  
Author(s):  
Christopher B. Raub ◽  
Chen-Chung Lee ◽  
Darryl Shibata ◽  
Clive Taylor ◽  
Emil Kartalov

1998 ◽  
Vol 28 ◽  
pp. 105
Author(s):  
Zs. Simon ◽  
G. Lotz ◽  
B. Nemes ◽  
F. Szalav ◽  
G. Lengyel ◽  
...  

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