Rapid quantification and in situ detection of nitrifying bacteria in biofilms by monoclonal antibody method

2000 ◽  
Vol 41 (4-5) ◽  
pp. 301-308 ◽  
Author(s):  
N. Noda ◽  
H. Ikuta ◽  
Y. Ebie ◽  
A. Hirata ◽  
S. Tsuneda ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Fluorescent Antibody ◽  
Nitrifying Bacteria ◽  
Fluorescent Antibody Technique ◽  
Antibody Technique ◽  
Antibody Method ◽  
In Situ Detection ◽  
Rapid Quantification

Fluorescent antibody technique by the monoclonal antibody method is very useful and helpful for the rapid quantification and in situ detection of the specific bacteria like nitrifiers in a mixed baxterial habitat such as a biofilm. In this study, twelve monoclonal antibodies against Nitrosomonas europaea (IFO14298) and sixteen against Nitrobacter winogradskyi (IFO14297) were raised from splenocytes of mice (BALB/c). It was found that these antibodies exhibited little cross reactivity against various kinds of heterotrophic bacteria. The direct cell count method using monoclonal antibodies could exactly detect and rapidly quantify N. europaea and N. winogradskyi. Moreover, the distribution of N. europaea and N. winogradskyi in a biofilm could be examined by in situ fluorescent antibody technique. It was shown that most of N. winogradskyi existed near the surface part and most of N. europaea existed at the inner part of the polyethylene glycol (PEG) gel pellet, which had entrapped activated sludge and used in a landfill leachate treatment reactor. It was suggested that this monoclonal antibody method was utilized for estimating and controlling the population of nitrifying bacteria as a quick and favorable tool.

Comparison of detection specificity of nitrifying bacteria in biofilm using fluorescence in situ hybridization and in situ fluorescent antibody methods

2003 ◽  
Vol 47 (5) ◽  
pp. 129-132
Author(s):  
N. Noda ◽  
Y. Ebie ◽  
M. Matsumura ◽  
S. Tsuneda ◽  
A. Hirata ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Fluorescent Antibody ◽  
Nitrifying Bacteria ◽  
Specific Fluorescence ◽  
Antibody Method ◽  
In Situ Detection ◽  
Detection Specificity ◽  
Fluorescent Antibody Method

The in situ fluorescent antibody and fluorescence in situ hybridization (FISH) methods are very useful in the in situ detection of specific bacteria like nitrifiers in a biofilm. In this study, simultaneous staining using the FISH and in situ fluorescent antibody methods was examined. As a result, no specific fluorescence was observed with either method when FISH was performed followed by the in situ fluorescent antibody method; however, when the in situ fluorescent antibody method was performed first followed by FISH, specific fluorescence was observed in both cases. Moreover, it was suggested that the detection specificities of FISH and the in situ fluorescent antibody method are almost identical.

STUDIES ON H. PERTUSSIS LIQUID CULTURES: III. LOCALIZATION OF SURFACE ANTIGENS BY MEANS OF FLUORESCENT ANTIBODY

1956 ◽  
Vol 2 (7) ◽  
pp. 677-683 ◽  
Author(s):  
J. de Repentigny ◽  
A. Frappier
Keyword(s):  
Fluorescent Antibody ◽  
Surface Antigens ◽  
Fluorescent Antibody Technique ◽  
Antibody Technique ◽  
Solid Media ◽  
Antigenic Properties ◽  
Antigenic Material ◽  
Capsular Antigens ◽  
The Relationship

The fluorescent antibody technique was used in an attempt to clarify the relationship between the morphology of the surface of Haemophilus pertussis and the antigenic properties of its capsular antigens. First, specific antibodies were produced by injection into rabbits of agglutinogenic and protective surface washings of the bacilli. Then, these antibodies were made fluorescent and used to mark specifically and in situ the original capsular antigens at the surface of bacilli grown in liquid as well as on solid media. Thus was obtained morphological and specific evidence for the presence of a capsule containing antigenic material.

scholarly journals Diagnosis of Avian Infectious Bronchitis by Indirect Fluorescent Antibody Technique Using Monoclonal Antibody

1993 ◽  
Vol 46 (2) ◽  
pp. 117-119 ◽  
Author(s):  
Kazuki ISHIBASHI ◽  
Hitomi SHIRAKAWA ◽  
Yoshifumi TOMISHITA ◽  
Hiroyuki MATUO ◽  
Akira WATANABE ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Fluorescent Antibody ◽  
Fluorescent Antibody Technique ◽  
Indirect Fluorescent Antibody ◽  
Antibody Technique ◽  
Infectious Bronchitis

Fasciola hepatica: development of monoclonal antibodies against somatic antigens and their characterization by ultrastructural localization of antibody binding

1988 ◽  
Vol 62 (1) ◽  
pp. 15-28 ◽  
Author(s):  
R. E. B. Hanna ◽  
A. G. Trudgett ◽  
A. Anderson
Keyword(s):  
Monoclonal Antibodies ◽  
Fasciola Hepatica ◽  
Fluorescent Antibody ◽  
Parenchymal Cell ◽  
Ultrastructural Localization ◽  
Immunogold Labelling ◽  
Antigenic Determinants ◽  
Fluorescent Antibody Technique ◽  
Adult Fluke ◽  
Antibody Technique

ABSTRACTA series of monoclonal antibodies was prepared against tegumental and internal antigens ofFasciola hepaticaby immunizing mice with whole adult-fluke homogenates prior to harvesting the splenic lymphocytes for fusion. Preliminary screening by the Indirect Fluorescent Antibody technique indicated the occurrence of discrete groups of monoclonals differing from one another in tissue-specificity but within which IFA labelling patterns were fairly consistent. Representative hybridomas for 5 of these groups were stabilized and used to produce ascites fluid in mice. By application of an immunogold labelling technique it was possible to map the distribution of antigens for which each monoclonal antibody had affinity throughout the tissues of 4-week and 12-week flukes. Several monoclonals specifically labelled antigenic determinants on the important tegumental antigen T1. However the distribution of gold colloid labelling suggested that epitopes other than that normally exposed to the infected host were recognized; and several monoclonals specifically attached to T1 antigen in the tegument of juvenile worms only. The glycocalyx of the gut and excretory system of flukes shared T1 antigenicity with the tegument. Monoclonal antibodies were produced against an internal immunogen associated with ribosomes and heterochromatin in active protein-producing cells, and against interstitial material of adult flukes. Monoclonals against antigens in parenchymal cell cytoplasm and in mature vitelline cells were recognized but the corresponding hybridomas were not stabilized.

Development of a rapid quantification method for nitrosomonas and nitrobacter using elisa for wastewater treatment facilities

1997 ◽  
Vol 36 (12) ◽  
pp. 169-174 ◽  
Author(s):  
Yuhei Inamori ◽  
Tomotake Takai ◽  
Naohiro Noda ◽  
Akira Hirata ◽  
Hiroshi Niioka ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Heterotrophic Bacteria ◽  
Competitive Elisa ◽  
Nitrifying Bacteria ◽  
Enzyme Linked Immunosorbent Assay ◽  
Quantification Method ◽  
Atcc Strain ◽  
Antibody Method ◽  
Treatment Facilities ◽  
Rapid Quantification

Enzyme-linked immunosorbent assay (ELISA) by use of monoclonal antibodies (MAbs) is very useful and helpful for the detection and quantification of the specific bacteria like nitrifiers in a mixed bacterial habitat. In this study, seven monoclonal antibodies were raised from splenocytes of mice(BALB/c) that are specific for the surface antigen of the two kinds of nitrifying bacteria. Three were directed against Nitrosomonas europaea (IFO 14298) and four were directed against Nitrobacter winogradskyi (IFO 14297). Cross-reactivities of MAbs against other strains of nitrifying bacteria as well as some kinds of representative heterotrophic bacteria in activated sludge and biofilm were checked to determine the usefulness of MAbs. It was found that there were some strain specificities between the same genera of IFO and ATCC strain. By means of a competitive ELISA, correlation curves for quantifying nitrifying bacteria were developed in a pure culture. It was found that this monoclonal antibody method could be used as a quick and powerful tool for estimating and controlling the population of nitrifying bacteria.

The rapid quantification and detection of nitrifying bacteria by using monoclonal antibody method

2000 ◽  
Vol 42 (3-4) ◽  
pp. 1-7 ◽  
Author(s):  
H. Ikuta ◽  
N. Noda ◽  
Y. Ebie ◽  
A. Hirata ◽  
S. Tsuneda ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Wastewater Treatment ◽  
Monoclonal Antibodies ◽  
Heterotrophic Bacteria ◽  
Sandwich Elisa ◽  
Domestic Wastewater ◽  
Nitrifying Bacteria ◽  
Bacterial Number ◽  
Domestic Wastewater Treatment ◽  
Antibody Method

Monoclonal antibodies against the two kinds of nitrifying bacteria Nitrosomonas europaea (IFO14298) and Nitrobacter winogradskyi (IFO14297) were raised and isotypes of these monoclonal antibodies, IgM and IgG1, were successfully obtained. Cross reactivities of these monoclonal antibodies against various kinds of representative heterotrophic bacteria turned out to be relatively low by competitive ELISA. In contrast, these monoclonal antibodies were very specific for nitrifying bacteria used as antigens. By means of sandwich ELISA using different isotype monoclonal antibodies such as IgM and IgG1, calibration curves were successfully developed for quantification of nitrifying bacteria. It was shown that the obtainable lower limit of quantification of N. europaea and N. winogradskyi were 7.0 × 106 N/ml and were 6.0 × 105 N/ml, respectively. Nitrifying bacteria in activated sludge of advanced domestic wastewater treatment johkaso were counted by sandwich ELISA and MPN methods. The bacterial number estimated by MPN method was lower than that estimated by sandwich ELISA. It was indicated that this monoclonal antibody method could be used as a quick and powerful tool for estimating and controlling the population of nitrifying bacteria in the advanced domestic wastewater treatment processes.

scholarly journals IMMUNOFLUORESCENT LOCALIZATION OF ORNITHINE AMINOTRANSFERASE IN RAT LIVER

1970 ◽  
Vol 18 (4) ◽  
pp. 264-267 ◽  
Author(s):  
PATRICIA C. BRENNAN ◽  
C. PERAINO ◽  
R. J. M. FRY ◽  
R. W. SWICK
Keyword(s):  
Fluorescent Antibody ◽  
Protein Diet ◽  
Fluorescent Antibody Technique ◽  
Ornithine Aminotransferase ◽  
Immunofluorescent Localization ◽  
Antibody Technique ◽  
Specific Fluorescence ◽  
Hepatic Cells ◽  
Standard Laboratory Diet

Ornithine aminotransferase (l-ornithine:2-oxoacid aminotransferase) (EC 2.6.1.13) was localized in the livers of rats fed diets differing in protein content with an indirect fluorescent antibody technique. The ornithine aminotransferase specific fluorescence in livers from rats fed a 60% protein diet was distributed throughout the cytoplasm of hepatic cells, and numerous fluorescent cells were observed throughout the lobules. Stained hepatic cells of rats fed a standard laboratory diet (24% protein) appeared to be less numerous and the fluorescence less intense than in rats fed the 60% diet. Livers from rats fed the 0% protein diet showed no specific fluorescence. The identification of ornithine aminotransferase in situ with the fluorescent antibody technique is compatible with measurements of the activity of enzyme extracted from the livers of rats fed similar diets.

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