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.

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.

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.

Detection of total bile acids in biological samples using an indirect competitive ELISA based on four monoclonal antibodies

2017 ◽  
Vol 9 (4) ◽  
pp. 625-633 ◽  
Author(s):  
Shuchen Liu ◽  
Yue Zhang ◽  
Baoping Qu ◽  
Gaofeng Qin ◽  
Jinjun Cheng ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Clinical Practice ◽  
Biological Samples ◽  
Competitive Elisa ◽  
Routine Clinical Practice ◽  
Enzyme Linked Immunosorbent Assay ◽  
Indirect Competitive Elisa ◽  
Different Types ◽  
Total Bile Acids

We investigated a newly developed indirect competitive enzyme-linked immunosorbent assay for the determination of 5 major components of TBA, which works efficiently in different types of biological samples, and may be suitable for routine clinical practice.

Monoclonal Antibody-Based Enzyme Linked Immunosorbent Assay of Aflatoxin B1, T-2 Toxin, and Ochratoxin A in Barley

1990 ◽  
Vol 73 (1) ◽  
pp. 71-76 ◽  
Author(s):  
Nannapaneni Ramakrishna ◽  
John Lacey ◽  
Alan A.G Candlish ◽  
John E Smith ◽  
Ian A Goodbrand
Keyword(s):  
Monoclonal Antibodies ◽  
Ochratoxin A ◽  
Aflatoxin B1 ◽  
Chloroform Extract ◽  
Barley Grain ◽  
Competitive Elisa ◽  
Water Soluble ◽  
Enzyme Linked Immunosorbent Assay ◽  
Simple Liquid ◽  
The Mean

Abstract Aflatoxin B1 (B1), T-2 toxin (T2), and ochratoxin A (OA) were assayed in a single extract from barley grain by using competitive enzyme linked immunosorbent assays (ELISAs) with monoclonal antibodies. B1 and T2 monoclonal antibodies were conjugated to horseradish peroxidase for direct competitive ELISA while an indirect competitive ELISA was used for OA determination. The competitive ELISA detected 0.1 ng/mL of B1f 10 ng/mL of T2, or 1 ng/mL of OA. Acetonitrile- 0.5% KCI-6% H2S04 (89 + 10 + 1 ) extracts of barley grain either were diluted 1:10 for direct assay or were subjected to a simple liquid-liquid cleanup procedure to concentrate the extract 10:1 before assay. For cleanup, water was added to the acetonltrile extract to partition water-soluble interfering substances, and then the mycotoxins were re-extracted with chloroform. The chloroform extract was evaporated to dryness and redissolved in Tris HCI buffer for ELISA. The mean recoveries from barley spiked with 4-60 ng/g of B1( 50-5000 ng/g of T2, and 5-500 ng/g of OA were, respectively, 93.8, 80.6, and 95.8%. The mean within-assay, Inter-assay, and subsample coefficients of variation by ELISA of barley grain colonized with toxigenic fungi were <12% for Bi and OA but as high as 17% forT2.

Distribution of nitrifying bacteria in a shallow stream

1997 ◽  
Vol 36 (8-9) ◽  
pp. 161-166 ◽  
Author(s):  
Ivana Jancarkova ◽  
Tove A. Larsen ◽  
Willi Gujer
Keyword(s):  
Field Experiments ◽  
Heterotrophic Bacteria ◽  
Pollution Source ◽  
Nitrifying Bacteria ◽  
Channel Water ◽  
River Bed ◽  
Hydraulic Conditions ◽  
Major Floods ◽  
Shallow Stream ◽  
Stream Bed

A project investigating the dynamics of self-purification processes in a shallow stream is carried out. Effects of the concentration gradient due to the distance to the pollution source, of hydraulic conditions in the river bed and of storm floods on the distribution of nitrifying bacteria were studied with the help of laboratory and field experiments. Nitrifiers density on the surface of the stream bed increased rapidly up to a distance of 300 m from the WWTP indicating possible competition of the nitrifiers with the heterotrophic bacteria close to the WWTP. Afterwards a slight decrease in the downstream direction was observed. In vertical profiles, higher bacterial densities were found at sites with rapid infiltration of channel water to the stream bed than at sites with no exchange between channel water and stream bed water or where stream bed water exfiltrated. A major flood event scoured the nitrifiers nearly totally from the surface of the river bed. Major floods belong so to the most dominant processes controlling self-purification in shallow streams. Minor floods, however, don't scour bacteria in the depth of the stream bed that could then be important for the self-purification processes.

scholarly journals Epitope Mapping of Monoclonal Antibodies to Calreticulin Reveals That Charged Amino Acids Are Essential for Antibody Binding

Antibodies ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 31
Author(s):  
Ann Christina Bergmann ◽  
Cecilie Kyllesbech ◽  
Rimantas Slibinskas ◽  
Evaldas Ciplys ◽  
Peter Højrup ◽  
...  
Keyword(s):  
Amino Acids ◽  
Monoclonal Antibodies ◽  
Epitope Mapping ◽  
Biological Function ◽  
Specific Binding ◽  
Enzyme Linked Immunosorbent Assay ◽  
Potential Therapeutic Target ◽  
Charged Amino Acids ◽  
Chaperone Protein ◽  
Terminal Amino

Calreticulin is a chaperone protein, which is associated with myeloproliferative diseases. In this study, we used resin-bound peptides to characterize two monoclonal antibodies (mAbs) directed to calreticulin, mAb FMC 75 and mAb 16, which both have significantly contributed to understanding the biological function of calreticulin. The antigenicity of the resin-bound peptides was determined by modified enzyme-linked immunosorbent assay. Specific binding was determined to an 8-mer epitope located in the N-terminal (amino acids 34–41) and to a 12-mer peptide located in the C-terminal (amino acids 362–373). Using truncated peptides, the epitopes were identified as TSRWIESK and DEEQRLKEEED for mAb FMC 75 and mAb 16, respectively, where, especially the charged amino acids, were found to have a central role for a stable binding. Further studies indicated that the epitope of mAb FMC 75 is assessable in the oligomeric structure of calreticulin, making this epitope a potential therapeutic target.

scholarly journals A Survey of G6 and G10 Serotypes of Group a Bovine Rotaviruses from Diarrheic Beef and Dairy Calves using Monoclonal Antibodies in ELISA

1994 ◽  
Vol 6 (2) ◽  
pp. 175-181 ◽  
Author(s):  
A. Lucchelli ◽  
S. Y. Kang ◽  
M. K. Jayasekera ◽  
A. V. Parwani ◽  
D. H. Zeman ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
South Dakota ◽  
Dairy Herd ◽  
Dairy Calves ◽  
Enzyme Linked Immunosorbent Assay ◽  
Positive Stool ◽  
Field Samples ◽  
Group A Rotaviruses ◽  
Beef Herds ◽  
Group A

Group A bovine rotaviruses (BRV) have been identified worldwide as a major cause of diarrhea in the young of many species, including humans. Group A rotaviruses are classified into serotypes on the basis of the outer capsid proteins, VP7 (G types) and VP4 (P types). To date, there are 14 G types of group A rotaviruses, with G1, G6, G8, and G10 described for BRV isolates. In this study, G6- and G lo-specific monoclonal antibodies (MAbs) were used in an enzyme-linked immunosorbent assay (ELISA) for the G typing of BRV-positive stool samples from diarrheic beef and dairy calves from South Dakota, Ohio, Michigan, Nebraska, and Washington, USA, and Ontario, Canada. ELISA plates were coated using a broadly reactive VP7 MAb (Common 60) or with G6- or G10-specific MAbs. BRV-positive fecal samples were diluted and added to duplicate wells, followed by the addition of polyclonal guinea pig anti-group A rotavirus serum as the secondary antibody. Several reference G6 and G10 BRV strains as well as other G types previously reported in cattle (G1, G2, G3, G8) and BRV-negative samples were included as G type specificity and negative controls. From a total of 308 field samples analyzed, 79% (244/308) tested positive by the broadly reactive VP7 MAb; of these, 54% (131/244) were G6 positive, 14% (35/244) were G10 positive, 4% (9/244) were both G6 and G10 positive, and 28% (69/244) were G6 and G10 negative. The negative samples may represent additional or undefined serotypes. The 89 samples from South Dakota were further subdivided into samples from beef ( n = 43) or dairy ( n = 46) herds. G6 was more prevalent in beef herd samples (67%) than in dairy herd samples (47.5%). In addition, dairy herds had higher percentages of G10-positive samples (17.5%) G6-G10 double positives (10%), and untypable samples (25%) than did beef herds, in which the prevalence of G10 positive samples was 5.5%, G6-G10 double positives was 5.5%, and untypable samples was 22%. Application of the serotype ELISA for the analysis of additional BRV samples will provide further epidemiologic data on the distribution of BRV serotypes in beef or dairy cattle, an important consideration for the development of improved BRV vaccines.

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