Himac cf 9rx

Blood samples were collected in BD Vacutainer tubes with clot activator and gel (BD Diagnostics, Plymouth, UK) from piglets and submitted to the Animal Diseases Diagnostic Center of National Pingtung University of Science and Technology, Taiwan. All piglets were divided into two groups, the MLV and E2 groups. Piglets in the MLV group were born from sows vaccinated with the CSFV MLV vaccine at 4 weeks post-farrowing, whereas piglets in the E2 group were born from sows vaccinated with the CSFV E2 subunit vaccine (Bayovac^® CSF-E2, Bayer Animal Health) at 4–5 weeks pre-farrowing. The blood samples were centrifuged at 2150× g for 15 min with a Himac CF 9RX (Hitachi Koki, Tokyo, Japan), and then the sera were carefully transferred into 1.5 mL centrifuge tubes. The stock serum was kept at −80 °C until needed.

Heparinized blood samples were centrifuged at 2150× g for 20 min at 4 °C. Samples of plasma supernatant were collected for biochemical analyses. Plasma CK and AST were measured using assay kits according to the manufacturer’s instructions. Sorbitol dehydrogenase (SDH) activities were measured as previously described [12 (link)].
Excised myocardial ventricular tissue samples were rinsed with ice-cold homogenizing buffer (50 mM Tris-HCl, 325 mM sucrose and 1 mM EDTA, pH 7.0). Liver tissue samples were rinsed with ice-cold homogenizing buffer (5 mM Tris-HCl, 250 mM sucrose and 0.1 mM EDTA, pH 7.0). Tissue homogenates were prepared by homogenizing ~0.5 g of minced tissue in 5 mL ice-cold homogenizing buffer (i.e., a 10% homogenate (w/v)) in a Teflon-in glass homogenizer (Glas-Col, Terre Haute, IN, USA) on ice at a speed of 40 rpm for 10 strokes. The homogenates were centrifuged (Himac CF 9RX, Hitachi Koki Co., Ltd., Ibaraki, Japan) at 600× g for 20 min at 4 °C. Mitochondrial fractions were obtained by centrifugation (Himac CR21G, Hitachi Koki Co., Ltd., Ibaraki, Japan) at 9200× g for 30 min at 4 °C.