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

2005 ◽  
Vol 25 (1) ◽  
pp. 4-8 ◽  
Author(s):  
Ronnie A. Assis ◽  
Francisco C.F. Lobato ◽  
Rogéria Serakides ◽  
Renato L. Santos ◽  
Guilherme R.C. Dias ◽  
...  
Keyword(s):  
Guinea Pigs ◽  
Fluorescent Antibody ◽  
Deae Cellulose ◽  
Fluorescein Isothiocyanate ◽  
Type A ◽  
Fluorescent Antibody Technique ◽  
Antibody Technique ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed

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.

scholarly journals Lymphoplasmacytic Meningoencephalitis and Neuronal Necrosis Associated With Parvoviral Infection in Cats

2019 ◽  
Vol 56 (4) ◽  
pp. 604-608 ◽  
Author(s):  
Anna Kokosinska ◽  
Grazieli Maboni ◽  
Kathleen M. Kelly ◽  
Alex Molesan ◽  
Susan Sanchez ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Brain Tissue ◽  
Quantitative Polymerase Chain Reaction ◽  
Fluorescent Antibody ◽  
Chain Reaction ◽  
Neuronal Necrosis ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Polymerase Chain ◽  
Formalin Fixed

Neurologic manifestations other than cerebellar hypoplasia are rarely associated with feline panleukopenia virus (FPV) infection in cats. Here the authors describe lymphoplasmacytic meningoencephalitis and neuronal necrosis in 2 cats autopsied after exhibiting ataxia and nystagmus. Gross changes consisted of cerebellar herniation through the foramen magnum, with flattening of cerebrocortical gyri and narrowing of sulci. Histologically, lymphoplasmacytic meningoencephalitis, extensive neuronal necrosis, and neuroaxonal degeneration with digestion chambers were present in the telencephalon and brain stem in both cats. Frozen brain tissue of both cats was positive for parvoviral antigen via fluorescent antibody testing, and formalin-fixed, paraffin-embedded tissue sections of brain were immunoreactive for parvovirus antigen and positive for parvoviral DNA on in situ hybridization. Frozen brain tissue from 1 case was positive for parvovirus NS1 and VP2 genes using conventional polymerase chain reaction, and subsequent DNA sequencing and phylogenetic analysis revealed that the viral strain was a FPV. Reverse transcription quantitative polymerase chain reaction on formalin-fixed, paraffin-embedded brain tissue revealed high levels of parvovirus in both cases, supporting an acute and active viral infection. Although rare, FPV infection should be considered in cases of lymphoplasmacytic meningoencephalitis and neuronal necrosis in cats.

scholarly journals A case study of rabies diagnosis from formalin-fixed brain material : short communication

2011 ◽  
Vol 82 (4) ◽  
Author(s):  
J. Coertse ◽  
L. H. Nel ◽  
C. T. Sabeta ◽  
J. Weyer ◽  
A. Grobler ◽  
...  
Keyword(s):  
South Africa ◽  
Rabies Virus ◽  
Fluorescent Antibody ◽  
Domestic Dog ◽  
Health Priorities ◽  
Fluorescent Antibody Technique ◽  
Rt Pcr ◽  
Antibody Technique ◽  
Formalin Fixed ◽  
Brain Tissues

Rabies is caused by several Lyssavirus species, a group of negative sense RNA viruses. Although rabies is preventable, it is often neglected particularly in developing countries in the face of many competing public and veterinary health priorities. Epidemiological information based on laboratory-based surveillance data is critical to adequately strategise control and prevention plans. In this regard the fluorescent antibody test for rabies virus antigen in brain tissues is still considered the basic requirement for laboratory confirmation of animal cases. Occasionally brain tissues from suspected rabid animals are still submitted in formalin, although this has been discouraged for a number of years. Immunohistochemical testing or a modified fluorescent antibody technique can be performed on such samples. However, this method is cumbersome and cannot distinguish between different Lyssavirus species. Owing to RNA degradation in formalin-fixed tissues, conventional RT-PCR methodologies have also been proven to be unreliable. This report is concerned with a rabies case in a domestic dog from an area in South Africa where rabies is not common. Typing of the virus involved was therefore important, but the only available sample was submitted as a formalin-fixed specimen. A real-time RT-PCR method was therefore applied and it was possible to confirmrabies and obtain phylogenetic information that indicated a close relationship between this virus and the canid rabies virus variants from another province (KwaZulu-Natal) in South Africa.

Fluorescent Antibody Studies in vitro on Ascaris suum Goeze, 1782

1962 ◽  
Vol 36 (3) ◽  
pp. 339-346 ◽  
Author(s):  
L. F. Taffs ◽  
A. Voller
Keyword(s):  
Indirect Method ◽  
Ascaris Suum ◽  
Fluorescent Antibody ◽  
Fluorescein Isothiocyanate ◽  
Normal Serum ◽  
Fluorescent Antibody Technique ◽  
Antibody Technique ◽  
Antigenic Sites ◽  
Third Stage

The immunology of Ascaris suum was studied in vitro using the fluorescent antibody technique. Fluorescein isothiocyanate labelling of globulin was used in the direct and indirect method and by a combination of both methods.Fluorescein labelled antibody was incorporated into precipitates at the mouth, excretory pore, anus and on the cuticle of third stage larvae.The possible intercuticular localization of fluorescent antibody in moulting larvae and fluorescence of the ends of broken larvae were further observed.Circumlarval precipitates did not form nor did the larvae fluoresce in normal serum.The results indicate (1) that antigenic sites are located at the natural orifices and cuticle and (2) that the dead internal tissues and possibly the moulting substance are also antigenic.

scholarly journals Use of an Immunoperoxidase Technique to Detect Equine Herpesvirus-1 Antigen in Formalin-Fixed Paraffin-Embedded Equine Fetal Tissues

1993 ◽  
Vol 5 (1) ◽  
pp. 12-15 ◽  
Author(s):  
Patricia C. Schultheiss ◽  
James K. Collins ◽  
Jane Carman
Keyword(s):  
Fluorescent Antibody ◽  
Immunoperoxidase Technique ◽  
Equine Herpesvirus ◽  
Specific Staining ◽  
Equine Herpesvirus 1 ◽  
Formalin Fixed Paraffin ◽  
Induced Abortions ◽  
Formalin Fixed Paraffin Embedded ◽  
Herpesvirus 1 ◽  
Formalin Fixed

An indirect immunoperoxidase (IP) procedure using the avidin-biotin-peroxidase complex detection technique was developed to detect viral equine herpesvirus-1 (EHV-1) antigen in formalin-fixed paraffin-embedded tissues from aborted equine fetuses. The procedure was applied to liver, lung, and other tissues from 20 cases of confirmed or suspected EHV-1 -induced abortions. Specific staining was observed in tissue sections from EHV-1-infected fetuses. Positive IP staining was present in tissues of 7 cases that were also positive by fluorescent antibody (FA) and virus isolation (VI) and that had typical histologic lesions. There was no IP staining in 7 cases that had no histologic lesions and negative FA and VI results. Five cases had typical histologic lesions and positive results in only 1 laboratory test; 3 were positive by VI and 2 by FA. Liver of 1 case was positive by IP, but tissues were too autolytic for other tests to be evaluated.

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