ASSOCIATIONS OF BIOCHEMICAL CHANGES AND MATERNAL TRAITS WITH MUTATION 1843 (C>T) IN THE RYR1 GENE AS A COMMON CAUSE FOR PORCINE STRESS SYNDROME
Popovski ZT, Tanaskovska B, Miskoska-Milevska E, Andonov S, Domazetovska S
*Corresponding Author: Professor Zoran T. Popovski, Ph.D., Faculty of Agriculture and Food Sciences, Bld “Aleksandar Makedonski,” bb PB 297, 1000 Skopje, Republic of Macedonia. Tel: +389-70-252-731. Fax: +389-2-3134-310. E-mail: zoran_popovski@yahoo.com
page: 75

MATERIALS AND METHODS

Blood was taken from 73 randomly chosen sows that were not exposed to stress. After the centrifugation step (1000 × g for 10 min. at room temperature), the plasma samples underwent further biochemical analyses, while the leucocytes served for DNA extraction. DNA extraction from leucocytes was done using a DNA purification kit (Cat. #A1125; Promega, Madison, WI, USA), according to the manufacturer’s instructions. Genotyping of PSS was performed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) by digestion of the amplified part of the ryr1 gene where the common 1843 (C>T) mutation occurs by digestion with HhaI restriction endonuclease, as previously described [18]. Biochemical analyses of nine biochemical parameters [bilirubin, Ca2+, creatinine, total protein, AP, aspartate aminotransferase (AST), alanine aminotransferase (ALT), CPK and lactate dehydrogenase (LDH)], were determined using an automatic analyzer (Cobas-Integra 700; Roche Diagnostics, Indianapolis, IN, USA). Quantitative records on newborn, stillborn and weaned piglets (piglets that became independent) required for the statistical analyses were obtained from the data kept on the farms. Statistical analysis for genotype distribution and allele frequencies were determined by individual and allele counting. For the quantitative parameters and productive traits, descriptive statistical parameters (mean, minimum, maximum and mean standard error (MSE) were calculated. Later, the differences in the genotypes were tested with the Student t-test. All statistical evaluations were performed using the Statistical Package for the Social Sciences (SPSS), version 6.1 for Windows Student Version (SPSS Inc, Chicago, IL, USA; 1994).



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