The European hare (Lepus europeaus) is one of the most important game animal in Europe. Since the last twenty years the stability of European populations has been progressively declining due to several causes, among them the occurrence of European Brown Hare Syndrome (EBHS), an highly contagious disease caused by a calicivirus, nowadays endemic in most European countries. As reported in another contribution to this conference, the first outbreaks of the disease in North Italy in the 1990s forced the adoption of serological surveillance in protected areas, in order to control the dynamics of hare populations in relation to the spread of the virus. Thereafter the periodical reoccurrence of cases imposed the adoption of a more complete program with the aim of monitoring the whole hare population, including those dwelling in hunting territories. In fact, hunting activity may facilitate the spread of the virus and also natal dispersal rates of animals, higher in hunting areas than in non-hunting areas, may contribute to the diffusion of the disease. In addition to the examination of carcasses of dead animals for viral detection, such monitoring activity takes advantage from serological survey i.e. by checking the presence of antibodies to EBHSV in both hares shot during hunting activity (from September to December) or those captured in restocking areas at the end of the hunting season (December-January) before being moved and released in hunting-free areas. Since different types of blood sampling may be adopted according to each situation, we planned to compare the serological titres obtained by testing with competitive enzyme-linked immunosorbent assay (cELISA) the “classical” serum from blood taken from ear vein with samples of blood dried onto blotting paper and also bloody samples taken directly from the heart cavities during necropsy. The major aim was to establish the utility of each sampling method for verifying the health status of hares and particularly to check the possibility to get data from low density areas. From 2005 to 2012 we analysed the following samples: a) serum from venous blood and blotting paper of 305 animals; b) blotting paper and cardiac blood of 182 animals; c) serum from venous blood and cardiac blood of 95 animals. Two small squares of approximately 6x6 mm of side were cut from dried blotters and placed in 100 μl of phosphate-buffered saline (pH 7,4) for 1 night and then 32μl of the eluted was recovered for ELISA testing. Hearts were collected by hunters during evisceration and delivered to the laboratory, where the bloody cardiac fluid (7 μl) was obtained by pressing the heart. Even if blotting paper and cardiac blood slightly underestimate the EBHSV antibody titres, if compared to the titre of serum from venous blood, both these “alternative” sampling methods may be useful for field studies. In particular blotting paper appears easier in sampling, storing and transporting also for hunters. In addition the slightly underestimates of antibody titres do not prevent to correctly interpret the sero-epidemiological results with regard to the understanding of spatial/time exposure of the population to EBHS and the ability of single hares to resist the EBHSV infection.

Comparison of differential sampling methods for serological tests applied to European Brown Hare Syndrome (EBHS) monitoring programs / M. Chiari, E. Gioia, N. Ferrari, L. Capucci, A. Lavazza - In: 4th World Lagomorph Conference[s.l] : World Lagomorph Society, 2012 Jul 23. - pp. 39-39 (( Intervento presentato al 4. convegno World Lagomorph Conference tenutosi a Wien nel 2012.

Comparison of differential sampling methods for serological tests applied to European Brown Hare Syndrome (EBHS) monitoring programs

M. Chiari
Primo
;
N. Ferrari;
2012

Abstract

The European hare (Lepus europeaus) is one of the most important game animal in Europe. Since the last twenty years the stability of European populations has been progressively declining due to several causes, among them the occurrence of European Brown Hare Syndrome (EBHS), an highly contagious disease caused by a calicivirus, nowadays endemic in most European countries. As reported in another contribution to this conference, the first outbreaks of the disease in North Italy in the 1990s forced the adoption of serological surveillance in protected areas, in order to control the dynamics of hare populations in relation to the spread of the virus. Thereafter the periodical reoccurrence of cases imposed the adoption of a more complete program with the aim of monitoring the whole hare population, including those dwelling in hunting territories. In fact, hunting activity may facilitate the spread of the virus and also natal dispersal rates of animals, higher in hunting areas than in non-hunting areas, may contribute to the diffusion of the disease. In addition to the examination of carcasses of dead animals for viral detection, such monitoring activity takes advantage from serological survey i.e. by checking the presence of antibodies to EBHSV in both hares shot during hunting activity (from September to December) or those captured in restocking areas at the end of the hunting season (December-January) before being moved and released in hunting-free areas. Since different types of blood sampling may be adopted according to each situation, we planned to compare the serological titres obtained by testing with competitive enzyme-linked immunosorbent assay (cELISA) the “classical” serum from blood taken from ear vein with samples of blood dried onto blotting paper and also bloody samples taken directly from the heart cavities during necropsy. The major aim was to establish the utility of each sampling method for verifying the health status of hares and particularly to check the possibility to get data from low density areas. From 2005 to 2012 we analysed the following samples: a) serum from venous blood and blotting paper of 305 animals; b) blotting paper and cardiac blood of 182 animals; c) serum from venous blood and cardiac blood of 95 animals. Two small squares of approximately 6x6 mm of side were cut from dried blotters and placed in 100 μl of phosphate-buffered saline (pH 7,4) for 1 night and then 32μl of the eluted was recovered for ELISA testing. Hearts were collected by hunters during evisceration and delivered to the laboratory, where the bloody cardiac fluid (7 μl) was obtained by pressing the heart. Even if blotting paper and cardiac blood slightly underestimate the EBHSV antibody titres, if compared to the titre of serum from venous blood, both these “alternative” sampling methods may be useful for field studies. In particular blotting paper appears easier in sampling, storing and transporting also for hunters. In addition the slightly underestimates of antibody titres do not prevent to correctly interpret the sero-epidemiological results with regard to the understanding of spatial/time exposure of the population to EBHS and the ability of single hares to resist the EBHSV infection.
Settore VET/05 - Malattie Infettive degli Animali Domestici
Settore VET/06 - Parassitologia e Malattie Parassitarie degli Animali
23-lug-2012
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/230986
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