ABSTRACT
The objective of this work was to investigate the use of a halothane to screen broiler chickens prone to develop PSE (Pale, Soft, Exudative) meat through a special gas chamber connected to a veterinarian anaesthetic apparatus developed in our laboratory. Anaesthesia was induced with approx. 3.0 percent halothane at a flow rate of 6.0 Lm-1 in pure oxygen for 5 min. Commercial male broilers (n=342) aged 46 days old were evaluated and classified as either halothane sensitive (HAL+) or insensitive (HAL-), depending on the leg rigidity response. Approximately 27.5 percent (n=94) of broilers were HAL+ and 72.5 percent (n=248) were HAL-. This is a simple and rapid technique to evaluate broiler sensitivity to halothane and identify broilers prone to develop PSE meat. The occurrence of PSE meat in HAL insensitive broiler chicken samples suggests that other factors are related to PSE occurrence.
O objetivo deste trabalho foi investigar o uso do halotano para avaliar e identificar frangos propensos ao desenvolvimento de carnes PSE (Pálida, Flácida, Exsudativa) por meio de uma câmara desenvolvida em nossos laboratórios. Esta foi conectada a um equipamento de anestesia veterinária e a anestesia foi então induzida com aproximadamente 3,0 por cento de halotano a um fluxo de 6,0 L/min em puro oxigênio. A sensibilidade de três frangos foi avaliada simultaneamente. Os frangos de linhagem comercial (n=352) com 46 dias de idade foram expostos à 3,0 por cento de halotano por 5 min e classificados como halotano sensível (HAL +) e não-sensível (HAL -), baseado no enrijecimento dos seus membros inferiores. Aproximadamente 27,5 por cento (n=94) frangos foram HAL+ e 72,5 por cento (n=248) HAL-. Amostras do músculo Pectoralis major (n=75) foram coletadas após o abate dos animais para as medidas de pH e cor, determinadas nos filés mantidos a 4 C por 24 h postmortem. Frangos HAL+ apresentaram maior porcentagem de carnes PSE quando comparadas ao grupo HAL-. Este é um teste simples e rápido para a avaliação da sensibilidade das aves ao halotano e identificar frangos propensos ao desenvolvimento de carnes PSE embora amostras de HAL - também apresentaram carnes PSE sugerindo que outros fatores estão relacionados com a formação da anormalidade.
ABSTRACT
Objective : The purpose of the presera study was to devélop a molecular genotyping method test by using a real time PCR hybridization probé and applying it to the analysis of C1843T mutations of the Sus scrofa RYR1 gene. Animáis population Three PSS-susceptible and PSS non-susceptible crossbred swine races were used for the experiments: Pietrain X Landrace Belga, Pietrain X Large White and Pietrain X Duroc. Methods: We have devéloped a genotyping method by using a hybridization probé and applied it to the analysis of C1843T mutations of the RYR1 gene, associated with PSS susceptibility. Genotyping results obtained by hybridization probé strategies were confirmed by restriction analysis and sequencing. In addi-tion, phenotype/genotype correlation analyses were devéloped by using the in vitro contracture test and confirmed the in vivo hálothane-succinylcholine challenge. Results: The real-time PCR with fluorescent hybridization probé methodology was designed to identify ho-mozygous PSS-resistant, PSS-susceptible animáis as well as heterozygous carriers. All cases genotyped by fluorescent hybridization probes were in agreement with PCR restriction enzyme digestión and sequencing and showed a 100% concordance between the in vivo and in vitro porcine stress syndrome (PSS) susceptibility results. Conclusions and clinical relevance: The real-time PCR with fluorescent hybridization probé method described here provides a rapid, easily interpretable and réliáble tool for genotyping the C1843T (Arg615-Cys) polymorphism of the RYR1 gene. This new methodology may be useful in the wide-scale genotyping of PSS-susceptibility and genetic selection.