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The Studies on Immuno-histochemical Techniques to Detect Riemerella Anatipestifer (RA) and Distribution of RA Antigen and Pathological in Artifical Infected Duckling

Author: LiuWeiPing
Tutor: ChengAnChun;WangMingShu
School: Sichuan Agricultural University
Course: Preventive Veterinary Medicine
Keywords: Riemerella Anatipestifer Immunohistochemistry Pathology Antigen distribution
CLC: S858.32
Type: Master's thesis
Year: 2003
Downloads: 164
Quote: 3
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Abstract


In this study successfully established indirect immunoperoxidase staining (Indirect Immunoperoxidase Staining Methods IISM) detection RA antigen, crossover study, absorption test, the alternative test positive, the negative control experiments show that the method has high sensitivity, specificity and good benefits. The methods work best conditions: fixed with 4% paraformaldehyde solution organization, sticky tablet APES (3-Amino Propyltriethoxy Silane), paraffin sections in 60 ℃ for 1h after indirect staining an anti-dilution of 1:25, the working concentration of the labeled secondary antibody (HRP) to 1:25,1% hydrochloric repair antigen 30min, 3% over the hydrogen peroxide methanol solution treated sample 20min, an anti-incubation time was 37 ℃ under 60min, HRP anti Cultivation time 37 ° C for the next 30min, blocking serum incubation time 37 ° C for the next 30min, a DAB (3,3 '-Diaminobenzidine tetrahydrochloride; 3,3' - two amino benzidine tetrahydrochloride) color 1min, satisfactory dyeing effect can be obtained. Successfully copied ducks source artificially infected ducklings after subcutaneous and intranasal two ways serotype 1 Anatipestifer labor cases, fibrinous pericarditis, perihepatitis and airbag Yandeng typical symptoms, artificially infected 6 hours after some ducks appear only tears symptoms after 12 hours, the desire to drink significantly decreased appetite waste must, volleyball Brown Xifen. 18 hours after the individual duck only neurological symptoms, mainly fell on the ground and can not anyway, drowsiness, head slightly swollen row of white Xifen, drowsiness, sleep like a nod general, sometimes shrill sharp call Head and Neck contraction, like chills, could not stand to crouch, stretched out a wing, cloudy eyes, blindness symptoms. A subcutaneous group of death, necropsy no obvious lesions. 24 hours microscopic examination found that some organs congestive bleeding. 36 hours after the pathological changes become evident, the pathological damage is more serious, subcutaneous and intranasal group began to die, which the subcutaneous group of mortality was peak-rise, the mortality rate was 40.9%, the highest. Intranasal group mortality was as high as 31.8%, after 50 hours, the subcutaneous group mortality begins, the mortality rate was 13.6%, while the the intranasal group's mortality peak of 50%. The subcutaneous group shorter incubation period (18 hours), the total mortality rate of 66.7%, the intranasal group long incubation period (48 hours), the total mortality rate as high as 95.5%. Necropsy appear the characteristic lesions the Riemerella Anatipestifer disease, the cellulose pericarditis perihepatitis, airsacculitis. Morbidity and mortality duck histological features as bleeding heart occurred edema, myocarditis, myocardial fibers dissolve disappear; liver bleeding, granular degeneration and vacuolar degeneration of liver cells, the final disintegration, necrosis; pulmonary hemorrhage, neutrophil infiltration, The lung room cell granular degeneration; renal hemorrhage, glomerular hyperplasia, tubule degeneration, lymphocyte and neutrophil granulocytes infiltration; intestinal the intestinal villi rupture, intestinal muscle rupture dissolved, midgut gland atrophy, intestinal villus epithelial cells vacuolar degeneration. Immune organs spleen hemorrhage, lymphopenia, granular degeneration, infiltration of neutrophils; bursal the mucosal epithelial degeneration off lymphopenia follicular atrophy, cortex, medulla, epithelial cell proliferation; thymus cellulose exudation, neutrophil infiltration. Brain lesions are less obvious, some vascular congestion, microglia nerve cell infiltration. The artificially infected ducklings liver and intestinal RA antigen detection rate of the highest average detection rate of duodenal highest (60.5%), the liver was 56.6%, 53.7% rectum cecum was 51.9%; lung, kidney and immune organs (spleen, bursa and thymus) average detection rate is also higher (45.7% lung, kidney 40%, 46.8%, spleen and bursa of 45%, the thymus was 41.7 pancreas, heart detection rate is relatively low (pancreas 39.4%, the Heart of 37.7%), brain and proventriculus detection rate is very low (less than 20%). antigen is widely distributed in the liver cells, intestinal epithelial cells, the cells of the midgut gland cells and tubular epithelial cells of the alveolar wall; immune organs spleen located in the white pulp lymphocytes, reticular cells, bursal lymphocytes located in lymphoid follicles medulla, thymus distributed in a corticomedullary lymphocytes; pancreatic gland cells and stromal cells distributed; brain distribution in the meninges, the the proventriculus is also a small amount of the distribution in the mucosa and submucosa cells. epithelial cells of RA antigen target cells. RA intracellular parasites.

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Livestock, poultry, wildlife diseases > Poultry > Duck
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