Immunohistochemical detection of Clostridia species in paraffin-embedded tissues of experimentally inoculated guinea pigs

Ronnie A. Assis, Francisco C F Lobato, Rogéria Serakides, Renato L. Santos, Guilherme R C Dias, Ricardo A P Nascimento, Vera L V Abreu, Patrícia M. Parreiras, Francisco A Uzal

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Blackleg is caused by Clostridium chauvoei, whereas malignant oedema is caused by C. chauvoei, C. septicum, C. sordellii, C. perfringens type A, and/or C. novyi type A. Anti-C. chauvoei, anti-C. septicum, anti-C. sordellii and anti-C. novyi type A polyclonal antibodies were produced in rabbits and purified in a column of DEAE-cellulose. Aliquots of the antisera were conjugated with fluorescein isothiocyanate and the remaining was used for the streptavidin biotin peroxidase technique (SBPT). SBPT was standardized to detect C. chauvoei, C. septicum, C. sordellii and C. novyi type A in formalin-fixed, paraffin-embedded tissues of guinea pigs. SBPT was compared to a fluorescent antibody technique (FAT). Sections and smears of muscle from inoculation area (MIA), heart, liver, spleen and kidney, were obtained for both SBPT and FAT. Cross-reactions between the different Clostridial species were not observed. C. chauvoei and C. septicum were detected in all specimens from the animals inoculated with these microorganisms, while only sections of muscle obtained from all the animals inoculated with C. sordellii and C. novyi type A were positive. The same results observed by the SBPT, were obtained on tissue smears of these microorganisms stained by the FAT. The results indicate that SBPT is suitable for detection of C. chauvoei, C. septicum, C. sordellii and C. novyi type A in formalin-fixed, paraffin-embedded tissues of guinea pigs.

Original languageEnglish (US)
Pages (from-to)4-8
Number of pages5
JournalPesquisa Veterinaria Brasileira
Volume25
Issue number1
StatePublished - Jan 2005

Fingerprint

Clostridium chauvoei
streptavidin
Streptavidin
Clostridium
biotin
Biotin
Paraffin
guinea pigs
alkanes
Peroxidase
peroxidase
Guinea Pigs
fluorescent antibody technique
Fluorescent Antibody Technique
formalin
Formaldehyde
blackleg (animal disease)
Clostridium sordellii
Clostridium novyi
Clostridium septicum

Keywords

  • Blackleg
  • Clostridia
  • Fluorescent antibody technique
  • Guinea pigs
  • Malignant oedema
  • Streptavidin biotin peroxidase technique

ASJC Scopus subject areas

  • veterinary(all)

Cite this

Assis, R. A., Lobato, F. C. F., Serakides, R., Santos, R. L., Dias, G. R. C., Nascimento, R. A. P., ... Uzal, F. A. (2005). Immunohistochemical detection of Clostridia species in paraffin-embedded tissues of experimentally inoculated guinea pigs. Pesquisa Veterinaria Brasileira, 25(1), 4-8.

Immunohistochemical detection of Clostridia species in paraffin-embedded tissues of experimentally inoculated guinea pigs. / Assis, Ronnie A.; Lobato, Francisco C F; Serakides, Rogéria; Santos, Renato L.; Dias, Guilherme R C; Nascimento, Ricardo A P; Abreu, Vera L V; Parreiras, Patrícia M.; Uzal, Francisco A.

In: Pesquisa Veterinaria Brasileira, Vol. 25, No. 1, 01.2005, p. 4-8.

Research output: Contribution to journalArticle

Assis, RA, Lobato, FCF, Serakides, R, Santos, RL, Dias, GRC, Nascimento, RAP, Abreu, VLV, Parreiras, PM & Uzal, FA 2005, 'Immunohistochemical detection of Clostridia species in paraffin-embedded tissues of experimentally inoculated guinea pigs', Pesquisa Veterinaria Brasileira, vol. 25, no. 1, pp. 4-8.
Assis, Ronnie A. ; Lobato, Francisco C F ; Serakides, Rogéria ; Santos, Renato L. ; Dias, Guilherme R C ; Nascimento, Ricardo A P ; Abreu, Vera L V ; Parreiras, Patrícia M. ; Uzal, Francisco A. / Immunohistochemical detection of Clostridia species in paraffin-embedded tissues of experimentally inoculated guinea pigs. In: Pesquisa Veterinaria Brasileira. 2005 ; Vol. 25, No. 1. pp. 4-8.
@article{e22b189906fa4eca8f35020f4c17b33b,
title = "Immunohistochemical detection of Clostridia species in paraffin-embedded tissues of experimentally inoculated guinea pigs",
abstract = "Blackleg is caused by Clostridium chauvoei, whereas malignant oedema is caused by C. chauvoei, C. septicum, C. sordellii, C. perfringens type A, and/or C. novyi type A. Anti-C. chauvoei, anti-C. septicum, anti-C. sordellii and anti-C. novyi type A polyclonal antibodies were produced in rabbits and purified in a column of DEAE-cellulose. Aliquots of the antisera were conjugated with fluorescein isothiocyanate and the remaining was used for the streptavidin biotin peroxidase technique (SBPT). SBPT was standardized to detect C. chauvoei, C. septicum, C. sordellii and C. novyi type A in formalin-fixed, paraffin-embedded tissues of guinea pigs. SBPT was compared to a fluorescent antibody technique (FAT). Sections and smears of muscle from inoculation area (MIA), heart, liver, spleen and kidney, were obtained for both SBPT and FAT. Cross-reactions between the different Clostridial species were not observed. C. chauvoei and C. septicum were detected in all specimens from the animals inoculated with these microorganisms, while only sections of muscle obtained from all the animals inoculated with C. sordellii and C. novyi type A were positive. The same results observed by the SBPT, were obtained on tissue smears of these microorganisms stained by the FAT. The results indicate that SBPT is suitable for detection of C. chauvoei, C. septicum, C. sordellii and C. novyi type A in formalin-fixed, paraffin-embedded tissues of guinea pigs.",
keywords = "Blackleg, Clostridia, Fluorescent antibody technique, Guinea pigs, Malignant oedema, Streptavidin biotin peroxidase technique",
author = "Assis, {Ronnie A.} and Lobato, {Francisco C F} and Rog{\'e}ria Serakides and Santos, {Renato L.} and Dias, {Guilherme R C} and Nascimento, {Ricardo A P} and Abreu, {Vera L V} and Parreiras, {Patr{\'i}cia M.} and Uzal, {Francisco A}",
year = "2005",
month = "1",
language = "English (US)",
volume = "25",
pages = "4--8",
journal = "Pesquisa Veterinaria Brasileira",
issn = "0100-736X",
publisher = "Colegio Brasileiro de Patologia Animal",
number = "1",

}

TY - JOUR

T1 - Immunohistochemical detection of Clostridia species in paraffin-embedded tissues of experimentally inoculated guinea pigs

AU - Assis, Ronnie A.

AU - Lobato, Francisco C F

AU - Serakides, Rogéria

AU - Santos, Renato L.

AU - Dias, Guilherme R C

AU - Nascimento, Ricardo A P

AU - Abreu, Vera L V

AU - Parreiras, Patrícia M.

AU - Uzal, Francisco A

PY - 2005/1

Y1 - 2005/1

N2 - Blackleg is caused by Clostridium chauvoei, whereas malignant oedema is caused by C. chauvoei, C. septicum, C. sordellii, C. perfringens type A, and/or C. novyi type A. Anti-C. chauvoei, anti-C. septicum, anti-C. sordellii and anti-C. novyi type A polyclonal antibodies were produced in rabbits and purified in a column of DEAE-cellulose. Aliquots of the antisera were conjugated with fluorescein isothiocyanate and the remaining was used for the streptavidin biotin peroxidase technique (SBPT). SBPT was standardized to detect C. chauvoei, C. septicum, C. sordellii and C. novyi type A in formalin-fixed, paraffin-embedded tissues of guinea pigs. SBPT was compared to a fluorescent antibody technique (FAT). Sections and smears of muscle from inoculation area (MIA), heart, liver, spleen and kidney, were obtained for both SBPT and FAT. Cross-reactions between the different Clostridial species were not observed. C. chauvoei and C. septicum were detected in all specimens from the animals inoculated with these microorganisms, while only sections of muscle obtained from all the animals inoculated with C. sordellii and C. novyi type A were positive. The same results observed by the SBPT, were obtained on tissue smears of these microorganisms stained by the FAT. The results indicate that SBPT is suitable for detection of C. chauvoei, C. septicum, C. sordellii and C. novyi type A in formalin-fixed, paraffin-embedded tissues of guinea pigs.

AB - Blackleg is caused by Clostridium chauvoei, whereas malignant oedema is caused by C. chauvoei, C. septicum, C. sordellii, C. perfringens type A, and/or C. novyi type A. Anti-C. chauvoei, anti-C. septicum, anti-C. sordellii and anti-C. novyi type A polyclonal antibodies were produced in rabbits and purified in a column of DEAE-cellulose. Aliquots of the antisera were conjugated with fluorescein isothiocyanate and the remaining was used for the streptavidin biotin peroxidase technique (SBPT). SBPT was standardized to detect C. chauvoei, C. septicum, C. sordellii and C. novyi type A in formalin-fixed, paraffin-embedded tissues of guinea pigs. SBPT was compared to a fluorescent antibody technique (FAT). Sections and smears of muscle from inoculation area (MIA), heart, liver, spleen and kidney, were obtained for both SBPT and FAT. Cross-reactions between the different Clostridial species were not observed. C. chauvoei and C. septicum were detected in all specimens from the animals inoculated with these microorganisms, while only sections of muscle obtained from all the animals inoculated with C. sordellii and C. novyi type A were positive. The same results observed by the SBPT, were obtained on tissue smears of these microorganisms stained by the FAT. The results indicate that SBPT is suitable for detection of C. chauvoei, C. septicum, C. sordellii and C. novyi type A in formalin-fixed, paraffin-embedded tissues of guinea pigs.

KW - Blackleg

KW - Clostridia

KW - Fluorescent antibody technique

KW - Guinea pigs

KW - Malignant oedema

KW - Streptavidin biotin peroxidase technique

UR - http://www.scopus.com/inward/record.url?scp=20044391824&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=20044391824&partnerID=8YFLogxK

M3 - Article

VL - 25

SP - 4

EP - 8

JO - Pesquisa Veterinaria Brasileira

JF - Pesquisa Veterinaria Brasileira

SN - 0100-736X

IS - 1

ER -