Brood stock segregation of spring Chinook salmon Oncorhynchus tshawytscha by use of the enzyme-linked immunosorbent assay (ELISA) and the fluorescent antibody technique (FAT) affects the prevalence and levels of Renibacterium salmoninarum infection in progeny

We developed a rapid method for detecting and quantifying the pathogen Renibacterium salmoninarum in coelomic fluid of spring chinook salmon (Oncorhynchus tshawytscha) by concentrating the bacteria on 0.2-μm polycarbonate filters and staining them with specific fluorescein-labeled antibody. Centrifugation of samples and resuspension of the sedimented material in phosphate-buffered saline containing Triton X-100 increased the ease of filtration. Background fluorescence was reduced by counterstaining filters with Eriochrome black T. Postfiltration staining, rinsing, and counterstaining were done in the syringe-mounted filter holders, reducing handling of the filters and possible loss of bacteria. The number of bacteria detected by the filtration – fluorescent antibody technique in a broth culture of R. salmoninarum ranged from 6.7 × 107to7.6 × 107/mL and was slightly higher than that determined by plate count (9.6 × 106/mL). Increasing the sample dilution or decreasing the number of microscope fields examined generally increased the variability of filter counts of R. salmoninarum. Using the filtration – fluorescent antibody technique, we detected the bacterium in the coelomic fluid of 85% of spawning female spring chinook salmon sampled from a hatchery population.

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Prevalence ofLeptospiraspp. in wild brown rats (Rattus norvegicus) on UK farms

Epidemiology and Infection ◽

10.1017/s0950268800052043 ◽

1995 ◽

Vol 114 (1) ◽

pp. 195-201 ◽

Cited By ~ 31

Author(s):

J. P. Webster ◽

W. A. Ellis ◽

D. W. Macdonald

Keyword(s):

Rattus Norvegicus ◽

Silver Staining ◽

Diagnostic Tests ◽

Fluorescent Antibody ◽

Agglutination Test ◽

Enzyme Linked Immunosorbent Assay ◽

Fluorescent Antibody Technique ◽

Antibody Technique ◽

Wild Rats ◽

The Uk

SUMMARYWild brown rats (Rattus norvegicus) are frequently implicated in the carriage and spread ofLeptospiraspp. Wild brown rats (n= 259) were trapped from 11 UK farms and tested forLeptospiraspp. using a number of diagnostic tests. The prevalence of leptospiral infection was low, but there was variation in the results obtained with the different diagnostic tests. Estimates of prevalence ranged between 0% by silver-staining of tissues, 1% by the microscopic agglutination test, 4% by the enzyme-linked immunosorbent assay, 4% by culture, and 8% by fluorescent antibody technique. In total, 37 (14%) rats were positive by at least one of the tests, which contrasts with the frequently reported prevalences of 50–70% for wild rats in the UK. Serovarbratislavawas as prevalent asicterohaemorrhagiae, although it was present only on farms with larger rat populations.

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In vitro methods for rabies virus detection, and evaluation of their use in the production of rabies immunoglobulin

BIOpreparations Prevention Diagnosis Treatment ◽

10.30895/2221-996x-2021-21-2-76-84 ◽

2021 ◽

Vol 21 (2) ◽

pp. 76-84

Author(s):

Yu. K. Gavrilova ◽

S. V. Generalov ◽

E. G. Abramova ◽

A. K. Nikiforov

Keyword(s):

Rabies Virus ◽

Fluorescent Antibody ◽

Enzyme Linked Immunosorbent Assay ◽

Fluorescent Antibody Technique ◽

Biological Method ◽

Culture Methods ◽

Antibody Technique ◽

Highly Sensitive ◽

Antibodies Detection

Current highly sensitive methods for rabies virus and rabies antibodies detection in biological material can be used not only for diagnosis and experimental research, but also for the production of antirabies medicines used for postexposure prophylaxis. The aim of the study was to analyse existing methods for rabies virus and rabies antibodies detection and to assess the potential for using these methods at the control stages during production of heterologous antirabies immunoglobulin obtained from equine serum. The search for cutting-edge highly sensitive in vitro control methods that could compete with the biological method, which is the main method used in antirabies immunoglobulin control, is an important prerequisite for improvement of the production technology and the quality of antirabies medicines. The study demonstrated that the following test methods can be used in the production of antirabies immunoglobulin: fluorescent antibody technique, enzyme-linked immunosorbent assay, cell culture methods, atomic force microscopy, and flow cytometry. These methods could be used alone or as an alternative to the biological method in white mice. These methods were chosen because of their high sensitivity, specificity, rapid and easy implementation, cost-effectiveness, and automatic recording of test results.

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Genetic effects of ELISA-based segregation for control of bacterial kidney disease in Chinook salmon (Oncorhynchus tshawytscha)

Canadian Journal of Fisheries and Aquatic Sciences ◽

10.1139/f06-163 ◽

2006 ◽

Vol 63 (12) ◽

pp. 2793-2808 ◽

Cited By ~ 13

Author(s):

Jeffrey J Hard ◽

Diane G Elliott ◽

Ronald J Pascho ◽

Dorothy M Chase ◽

Linda K Park ◽

...

Keyword(s):

Kidney Disease ◽

Chinook Salmon ◽

Oncorhynchus Tshawytscha ◽

Enzyme Linked Immunosorbent Assay ◽

Antigen Level ◽

Renibacterium Salmoninarum ◽

Listonella Anguillarum ◽

Bacterial Kidney Disease ◽

Genetic And Phenotypic Correlations ◽

High Heritability

We evaluated genetic variation in ability of Chinook salmon (Oncorhynchus tshawytscha) to resist two bacterial pathogens: Renibacterium salmoninarum, the agent of bacterial kidney disease (BKD), and Listonella anguillarum, an agent of vibriosis. After measuring R. salmoninarum antigen in 499 adults by enzyme-linked immunosorbent assay (ELISA), we mated each of 12 males with high or low antigen levels to two females with low to moderate levels and exposed subsets of their progeny to each pathogen separately. We found no correlation between R. salmoninarum antigen level in parents and survival of their progeny following pathogen exposure. We estimated high heritability for resistance to R. salmoninarum (survival h2 = 0.890 ± 0.256 (mean ± standard error)) independent of parental antigen level, but low heritability for resistance to L. anguillarum (h2 = 0.128 ± 0.078). The genetic correlation between these survivals (rA = –0.204 ± 0.309) was near zero. The genetic and phenotypic correlations between survival and antigen levels among surviving progeny exposed to R. salmoninarum were both negative (rA = –0.716 ± 0.140; rP = –0.378 ± 0.041), indicating that variation in antigen level is linked to survival. These results suggest that selective culling of female broodstock with high antigen titers, which is effective in controlling BKD in salmon hatcheries, will not affect resistance of their progeny.

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Comparison of the enzyme-linked immunosorbent assay (ELISA) and the fluorescent antibody test (FAT) for measuring the prevalences and levels of Renibacterium salmoninarum in wild and hatchery stocks of salmonid fishes in Alaska. USA

Diseases of Aquatic Organisms ◽

10.3354/dao016181 ◽

1993 ◽

Vol 16 ◽

pp. 181-189 ◽

Cited By ~ 20

Author(s):

TR Meyers ◽

S Short ◽

C Farrington ◽

K Lipson ◽

HJ Geiger ◽

...

Keyword(s):

Immunosorbent Assay ◽

Fluorescent Antibody ◽

Antibody Test ◽

Enzyme Linked Immunosorbent Assay ◽

Salmonid Fishes ◽

Renibacterium Salmoninarum

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Comparison of the Membrane-Filtration Fluorescent Antibody Test, the Enzyme-Linked Immunosorbent Assay, and the Polymerase Chain Reaction to Detect Renibacterium Salmoninarum in Salmonid Ovarian Fluid

Journal of Veterinary Diagnostic Investigation ◽

10.1177/104063879801000111 ◽

1998 ◽

Vol 10 (1) ◽

pp. 60-66 ◽

Cited By ~ 34

Author(s):

Ronald J. Pascho ◽

Dorothy Chase ◽

Connie L. McKibben

Keyword(s):

Polymerase Chain Reaction ◽

Immunosorbent Assay ◽

Membrane Filtration ◽

Fluorescent Antibody ◽

Antibody Test ◽

Enzyme Linked Immunosorbent Assay ◽

Ovarian Fluid ◽

Chain Reaction ◽

Renibacterium Salmoninarum ◽

Polymerase Chain

Ovarian fluid samples from naturally infected chinook salmon ( Oncorhynchus tshawytscha) were examined for the presence of Renibacterium salmoninarum by the membrane-filtration fluorescent antibody test (MF-FAT), an antigen capture enzyme-linked immunosorbent assay (ELISA), and a nested polymerase chain reaction (PCR). On the basis of the MF-FAT, 64% (66/103) samples contained detectable levels of R. salmoninarum cells. Among the positive fish, the R. salmoninarum concentrations ranged from 25 cells/ml to 4.3 × 109 cells/ml. A soluble antigenic fraction of R. salmoninarum was detected in 39% of the fish (40/103) by the ELISA. The ELISA is considered one of the most sensitive detection methods for bacterial kidney disease in tissues, yet it did not detect R. salmoninarum antigen consistently at bacterial cell concentrations below about 1.3 × 104 cells/ml according to the MF-FAT counts. When total DNA was extracted and tested in a nested PCR designed to amplify a 320-base-pair region of the gene encoding a soluble 57-kD protein of R. salmoninarum, 100% of the 100 samples tested were positive. The results provided strong evidence that R. salmoninarum may be present in ovarian fluids thought to be free of the bacterium on the basis of standard diagnostic methods.

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T77. ASSOCIATION BETWEEN INTRACELLULAR INFECTIOUS AGENTS AND SCHIZOPHRENIA

Schizophrenia Bulletin ◽

10.1093/schbul/sbaa029.637 ◽

2020 ◽

Vol 46 (Supplement_1) ◽

pp. S261-S261

Author(s):

Young-Joon Kwon ◽

Bo-Hyun Yoon ◽

Kwanghun Lee ◽

Moon-Doo Kim ◽

Beomwoo Nam ◽

...

Keyword(s):

Schizophrenia Patient ◽

Clinical Symptoms ◽

Fluorescent Antibody ◽

Chlamydia Infection ◽

Enzyme Linked Immunosorbent Assay ◽

Fluorescent Antibody Technique ◽

Igg Antibody ◽

Antibody Technique ◽

General Psychopathology ◽

Control Subjects

Abstract Background A number of studies have reported association between Toxoplasma gondii (T. gondii) and Chlamydia infection and the risk of schizophrenia. The aim of the present study was to compare the prevalence of T. gondii and Chlamydia infection between the schizophrenia and normal control subjects and to compare the clinical features between seropositive and seronegative schizophrenia patients. Methods The rate of serum reactivity to T. gondii, Chlamydia trachomatis (C. trachomatis), Chlamydia pneumonia in 96 schizophrenia and 50 control subjects was investigated using enzyme-linked immunosorbent assay and indirect fluorescent antibody technique. The clinical symptoms of the schizophrenia patients were scored with Positive and Negative Syndrome Scale and a comparative analysis was carried out. Results A significant positive association between immunoglobulin G (IgG) antibodies to T. gondii and C. trachomatis in schizophrenia was found, and the odds ratio of schizophrenia associated with IgG antibody was found to be 3.22 and 2.86, respectively. The Toxoplasma-seropositive schizophrenia patient had higher score on the negative subscale N1 and N7 and general psychopathology subscale G13, while C. trachomatis-seropositive schizophrenia patient had higher score on the general psychopathology subscale G10. Discussion The results from the present study suggest significant association between T. gondii, C. trachomatis infection and schizophrenia. In future, further studies are needed to elucidate the correlation between the two types of infection and schizophrenia.

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Comparison of two fluorescent antibody techniques (FATs) for detection and quantification of Renibacterium salmoninarum in coelomic fluid of spawning chinook salmon Oncorhynchus tshawytscha

Diseases of Aquatic Organisms ◽

10.3354/dao030037 ◽

1997 ◽

Vol 30 ◽

pp. 37-43 ◽

Cited By ~ 8

Author(s):

DG Elliott ◽

CL McKibben

Keyword(s):

Chinook Salmon ◽

Oncorhynchus Tshawytscha ◽

Fluorescent Antibody ◽

Coelomic Fluid ◽

Renibacterium Salmoninarum ◽

Detection And Quantification

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

Water Science & Technology ◽

10.2166/wst.2000.0459 ◽

2000 ◽

Vol 41 (4-5) ◽

pp. 301-308 ◽

Cited By ~ 1

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.

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