• Korean Journal of Veterinary Service
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• v.20 no.2
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• pp.195-203
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• 1997

Fecal samples were collected from 47 calves with diarrhea and 12 clinically normal co-h-abitants, and tested for virus using MDBK cell cultures. Three cytopathic viruses were isolated from 8 fecal samples obtained from diarrheic calves. The isolated viruses were neutralized by bovine coronavirus hyperimmune serum In plaque reduction assay and were detected in the cytoplasm of MDBK cell by bovine coronavirus hyperimmune serum using immunofluorescence staining. The viruses agglutinated mouse erythrocytes only among the various animal erythrocytes tested and new isolates were identified as bovine coronavirus.

• Korean Journal of Veterinary Service
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• v.21 no.3
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• pp.313-323
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• 1998

In order to establish a sensitive and specific diagnostic method for detection of antibody to Salmonella pullorum, a enzyme-linked immunosorbent assay(ELISA) was designed and standardized. The diagnostic efficacy of the established ELISA was compared with that of the serum plate agglutination test and immunodiffusion test for pullorum disease. 1. The chicken hyperimmune sera to Salmonella pullorum, S gallinarum, S typhimurium and S typhi were shown the cross reaction to S pullorum antigen by serum plate agglutination test. 2. When compared the cross reaction titer of microplate agglutination test for chickens hyperimmune sera, it was found that the titer were 64 in S pullorum, 32 in S gallinarum, 4 in S typhimurium and 8 in S typhi, respectively. 3. When compared the specificity of various antigen(HA, EA, PA and SA) by the immunodiffusion test, the most suitable antigen was phenol-treated bactrium. 4. The optimal concentration of S pullorum antigen for ELISA was 1 : 160 dilution of bacterium. 5. The efficacy of the ELISA for detection of S pullorum antibody was compared with serum Plate agglutination test and immunodiffusion test in chickens infected with S pullorum. The antibody was first detected at 6 days after infection using three tests examined. The antibody was alldetected at 9 days by ELISA, at 12 days by serumplate agglutination test, at 15 days by immunodiffusion test.

• Journal of Pharmacopuncture
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• v.25 no.2
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• pp.114-120
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• 2022

Objectives: Antivenom serums have been used extensively for over a century and are the only effective treatment option for snake bites and other dangerous animal envenomations. In therapeutic serum centers, a wide range of antivenoms is made from animal serum, mainly equine and sheep, that are immunized with single or multiple venoms. This work aimed to use caprylic acid (CA) to purify therapeutic snake antivenom. Methods: Plasma was obtained from equine immunized with a mixture of venoms. Immunized plasma was obtained by precipitation of different concentrations (2-5%) of CA. This methodology was compared to that based on ammonium sulfate (AS) precipitation. Sediment plasma proteins were purified by ion-exchange chromatography. Protein assay, SDSPAGE, and agar gel diffusion were performed. Results: The total protein precipitation with AS was higher than precipitation with CA, but the best results were obtained when CA was added to the plasma until a final CA concentration of 5% was reached. Chromatography and electrophoresis indicated a stronger band for the 5% CA, and the gel diffusion assay showed antigen-antibody interaction in the purified serum. Conclusion: The use of CA compared to the routine method for purifying hyperimmune serums is a practical and cost-effective method for preparing and producing therapeutic serums. It constitutes a potentially valuable technology for alleviating the critical shortage of antivenom in Iran.

• Korean Journal of Veterinary Research
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• v.29 no.4
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• pp.503-515
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• 1989

Critical parameters affecting sensitivity and specificity of an enzyme-linked immunosorbent assay(ELISA) for detection of antibodies to avian infections bronchitis virus(IBV) were standardized. By adopting the optimized conditions an equation calculating ELISA antibody titers from the observations at single serum dilution was formulated. The purified antigen of IBV Mass-41 strain was dispensed into polystyrene microplate wells at a concentration of 300ng per well($100{\mu}l$) and the plates were coated by completey drying at $37^{\circ}C$. Diluted chicken serum and horseradish peroxidase conjugated goat anti-chicken IgG were added in order in $100{\mu}l$ volumes per well and allowed to react for 30 minutes each at room temperature. Just before use and after each reaction the plates were washed three times with distilled water. Finally o-phenylenediamine solution was added as an enzyme substrate. After incubation for another 15 minutes at room temperature absorbances were read at 492nm. Hyperimmune serum against Mass-41 strain was used as internal reference positive(IRP) serum. After repeated titration of IRP and negative serum, a constant titer of IRP was determined. Serum titrations were carried out for various sample sera together with IRP and negative sera and the observed titers of sample sera were corrected by reflecting the ratio between observed and constant titers of IRP serum. These corrected titers of the sample sera were plotted against sample/positive(S/P) OD ratios. All the OD's measured in the serum titrations were also corrected by substracting negative serum OD. The following equation was formulated from the above data; $Log_{10}$ ELISA titer=$5.568({\log}_{10}S/P)+4.161$ Thus it was possible to calculate ELISA titer by measuring absorbance at 1/400 single serum dilution. Titer measured by cross ELISA tests employing Mass-41 strain and three local IBV isolates were similar. These results suggest that the ELISA tests standardized in this study can be used for evaluating not only vaccinal immunity but also for infection status against fields IBV's.

• Parasites, Hosts and Diseases
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• v.36 no.2
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• pp.127-132
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• 1998

Immunotherapy has been used in support of trimethoprim-sulfamethoxazole treatment for Pneumocvstis corinii pneumonia. The present study investigated the therapeutic or preventive effects of heterogeneous hyperimmune IgG antibody (HIA) in experimental rats. Their immunity was suppressed by steroid injection, and they were also injected peritoneally with HIA which reacted with 40-55, 92, 116, and 200 kDa bands of the crude antigen. All rats were infected by p. ccrinii and the cystic forms on lung impression smears were counted. The count was 20.5-76.5 (mean 52.5 ± 19.)1 in those which received steroid only, but decreased to 6.0-21.0 (mean 13.5 : 10.6) in those of group 3 which received HIA for the same duration. In other groups, the mean count ranged from 29.9 t 32.9 to 54.1 t 47.7, and in those which received 13.7 mg HIA the reduction effect was greater than in those which received 6.8 mg or 20.5 mg HIA. The present finding confirmed that in rats during the early stage of infection, the heterogeneous HIA to MSG antigen bands had a partial effect on p. cori,nii pneumonia, both prophylactically and therapeutically.

• Korean Journal of Veterinary Research
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• v.28 no.1
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• pp.89-97
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• 1988

Twenty-eight porcine rotavirus were isolated from piglets with diarrhoea in chonnam province. According to the age, 41 to 60 day old pigs showed the highest isolation frequency among the post weaning pigs. The characteristics of the field isolates were determined by electronmicroscopy(EM), immunofluorescent assay(FA), and electrophoretic migration patterns of the genome profiles. Some of the isolates showed remarkable haemagglutination activity against rabbit and dog erythrocytes, ranged from 4 to 2848, respectively. At least 3 serotypes of porcine rotavirus were recognized by serum neutralization test using serotype specific rotavirus hyperimmune sera.

• Parasites, Hosts and Diseases
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• v.44 no.4 s.140
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• pp.271-283
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• 2006

The third-stage larvae (L3) of the parasitic nematode, Anisakis simplex, have been implicated in the induction of hyperimmune allergic reactions in orally infected humans. In this work, we have conducted a review of an investigation into immune reactions occurring in animals experimentally infected with A. simplex L3. The patterns of serum antibody productions if the experimental animals against excretory-secretory products (ESP) of A. simplex L3 contributed to our current knowledge regarding specific humoral immune reactions in humans. In our review, we were able to determine that L3 infection of experimental animals may constitute a good model system for further exploration of immune mechanisms and allergy in anisakiasis of humans.

• Parasites, Hosts and Diseases
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• v.34 no.4
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• pp.239-246
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• 1996

The usefulness of malaria diagnosis by Plusmodium JaLcipawn-GDH (NADP+), obtained by affinity chromatography. is demonstrated in ELISA assays, testing IgG antibodies against GDH (NADP+) from patients with acute malaria, who have had two or more episodes of malaria. or from sera of hyperimmune patients. GDH (NADP+) thermal stability was demonstrated in a high heat resistance assay. The immunofluorescence assay demonstrated that anti-culture (P. falciporum) supernatant serum and anti-GDH (NADP+) of Proton app. recognized epitopes in Venezuelan isolates and Colombian and Brazilian malarial strains. The antigen is soluble, with high specificity is a potent imnlunogen and is thermoresistant. Key words: antigenic enzymes. glutamate dehydrogenase, malaria diagnosis, Plasmodium berghei, Plcswlodium ccthemelum, PlusmoniumJnlcipnmm, Plosmonium uiuox. soluble antigens.

• Parasites, Hosts and Diseases
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• v.40 no.1
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• pp.59-64
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• 2002

A Cryptosporidium parvum sporozoite and oocyst λgt11 cDNA library was screened with a hyperimmune rabbit serum that was developed against insoluble fragments of ultrasonicated oocysts. A clone named Cp22.4.1 encoding a protein of 231 amino acids with 4 zinc-finger domains characterized by a Cys-X2-Cys-X4-His-X4-Cys motif was isolated and characterized. There was a complete match between the sequencing data of the coding region of Cp22.4.1 and the corresponding gene at chromosomal level. Cloning in a pBAD-TOPO-TA expression vector permitted to evaluate the antigenicty of the recombinant His-tagged antigen. This antigen was recognized by 2 out of 5 sera from Cruptosporidium immune calves and not by sera from parasite naive animals.

• Korean Journal of Veterinary Research
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• v.40 no.1
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• pp.86-93
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• 2000

Paratuberculosis (Johne's disease), a chronic enteritis produced by Mycobacterium paratuberculosis, affects a large proportion of ruminants in all continents and causes important economic losses. The identification of well-characterized and species-specific components of M paratuberculosis would provide the means to improve the specificity and sensitivity of immunodiagnostic assays for Johne's disease. The aims of this study were to express the recombinant C-terminal of 34kDa protein (rC34P) of M paratuberculosis in E coli and to investigate the effectiveness of this protein in detecting antibodies to the native protein in sera from paratuberculosis infected cattle. The C-terminal of the gene encoding the 34kDa protein was amplified by polymerase chain reaction from the chromosomal DNA of M paratuberculosis (ATCC 19698) and cloned into vector pGEX-4T-2. Then, cloned plasmid was transformed into E coli DH5${\alpha}$ and the rC34P was overexpressed. The rC34P was purified by affinity chromatography and gel filtration. The rC34P was examined antigenicity by Western blot. The rC34P was reactive with culture positive bovine serum and hyperimmune rabbit anti-M paratuberculosis serum but was not reactive with culture negative bovine serum and tuberculin positive bovine serum in Western blot. In conclusion, the rC34P produced in this study is expected as a useful candidate for antigen in serological diagnosis of Johne's disease.