Samples of Pectoralis major m. were collected, and an RT-PCR analysis of the a-Ryanodine receptor (a RYR) from chicken mRNA hotspot region spanning aminoacid residues 386 to 540, numbered according to the turkey sequence, revealed two classes of transcripts. The sequences of the first class were similar to turkey and human with 97% and 74% of identity, respectively, and included all transcripts with substitutions in the nucleotide sequence. The second class was characterized by the deletion of nucleotides, leading to a premature stop codon and coding for a truncated and nonfunctional protein. These results are to date the first report related to the sequencing of the chicken αRYR hotspot region 1, which will possibility serve as a guide for further studies regarding a solution in the poultry production chain related to the problem of pale, soft and exudative (PSE) meat.

The biological cause of Pork Stress syndrome, which leads to PSE (pale, soft, exudative) meat, is excessive release of Ca(2+) ions, which is promoted by a genetic mutation in the ryanodine receptors (RyR) located in the sarcoplasmic reticulum of the skeletal muscle cells. We examined the relationship between the formation of PSE meat under halothane treatment and heat stress exposure in chicken alphaRYR hot spot fragments. Four test groups were compared: 1) birds slaughtered without any treatment, i.e., the control group (C); 2) birds slaughtered immediately after halothane treatment (H); 3) birds slaughtered immediately after heat stress treatment (HS), and 4) birds exposed to halothane and to heat stress (H+HS), before slaughtering. Breast muscle mRNA was extracted, amplified by RT-PCR, and sequenced. PSE meat was evaluated using color determination (L* value). The most common alteration was deletion of a single nucleotide, which generated a premature stop codon, resulting in the production of truncated proteins. The highest incidence of nonsense transcripts came with exposure to halothane; 80% of these abnormal transcripts were detected in H and H+HS groups. As a consequence, the incidence of abnormal meat was highest in the H+HS group (66%). In HS, H, and C groups, PSE meat developed in 60, 50, and 33% of the samples, respectively. Thus, halothane apparently modulates alphaRYR gene expression in this region, and synergically with exposure to heat stress, causes Avian Stress syndrome, resulting in PSE meat in broiler chickens.

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ABSTRACT

This work aimed to evaluate female lineage broilers for halothane sensitivity and for their susceptibility to the subsequent development of PSE meat. The halothane test was carried out in an anesthetic chamber with 3.0% halothane. The unconscious birds were examined for leg muscle rigidity. If one or both legs became extended and rigid, the birds were classified as halothane sensitive (HAL+), while unresponsive birds were classified as halothane negative (HAL-). The results showed that of 298 birds aged 42 days old, 95.6% were HAL- and 4.4% were HAL+. A sample of pectoralis major muscle was collected from HAL- (n=105) and HAL+ (n=13) birds. The pH and breast fillet color were determined at 4ºC, 24 hours post-mortem. Interestingly, only 2.5% of HAL+ birds displayed PSE meat characteristics compared to 12.7% of HAL- individuals. The halothane test demonstrated that female lineage broilers displayed very little sensitivity towards halothane, indicating that the development of PSE meat is related to other environmental factors.
O objetivo deste trabalho foi avaliar a sensibilidade de aves de uma linhagem fêmea ao halotano e sua relação com o desenvolvimento de carnes PSE. O teste do halotano foi conduzido com o auxílio de uma câmara anestésica com 3,0% de halotano volatilizado. As aves inconscientes foram examinadas pelo enrijecimento dos seus membros inferiores. Quando ambos ou um dos membros permaneceram rígidos, os frangos foram classificados como sensíveis ao halotano (HAL+) e os frangos sem enrijecimento dos membros foram classificados como não-sensíveis (HAL-). Os resultados mostraram que de 298 frangos com 42 dias de idade, 95,6% foram HAL-, e apenas 4,4% HAL+. O peito foi coletado das aves HAL- (n=105) e HAL+ (n=13) em que o pH e Cor (L*,a*,b*) foram determinados a 4ºC, 24h postmortem. Interessantemente, apenas 2,5% das aves HAL+ demonstraram carnes PSE, enquanto que as aves HAL- apresentaram 12,7% de carnes PSE, em relação ao total de aves abatidas. O teste do halotano demonstrou que frangos da linhagem fêmea mostraram pouquíssima sensibilidade ao halotano, indicando que a ocorrência de carnes PSE está mais associada a fatores ambientais