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
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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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