Establishment of a quantitative bovine CXCL8 sandwich ELISA with newly developed monoclonal antibodies.

Three IgG class anti-bovine CXCL8 (bCXCL8) monoclonal antibody (mAb)-secreting hybridomas, SH8-8D7, SH8-12A5 and SH8-2A1, were developed. SH8-8D7 was IgG2a, and SH8-12A5 and SH8-2A1 were IgG1. All three mAbs detected recombinant bCXCL8 (rbCXCL8) by immunoprecipitation and Western blotting. SH8-2A1 could neutralise the chemotactic activity of rbCXCL8 towards neutrophils. The quantitative bCXCL8 ELISA was constituted by the combination of SH8-12A5 and biotin-SH8-2A1. The detection range was 20-1000  pg/mL. A sandwich ELISA was used to measure native bCXCL8 derived from the supernatant of cultured bovine peripheral blood mononuclear cells stimulated with ConA, LPS or PHA. Furthermore, SH8-2A1 could detect bCXCL8 in formalin-fixed, paraffin-embedded, pneumonic calf tissues. These findings indicate that the newly developed anti-bCXCL8 mAbs could contribute to research on bovine inflammatory responses and immunology.

The effect of experimental inhibition of gastric acid secretion on curd formation in abomasum and weight gain of calves.

Eight Holstein bull calves were divided into two groups; a non-treated control group and a famotidine treated group. Fresh milk was fed twice a day. The experiment was conducted between 7 and 14 days of age. During the experimental period the control group was injected with physiological saline, and the famotidine group was injected with famotidine, a histamine-H2-receptor blocker, into the jugular vein 30 minutes prior to each feeding. The control group showed maximum curd formation 2 h after feeding at both 7 and 14 days of age. Curd scores of 7-day-old and 14-day-old calves were significantly lower in the famotidine than in the control group at 2 and 4 h after feeding. Most fecal samples from the famotidine group exhibited an acidic smell. The famotidine group showed significantly lower values for both average weight gain and the rate of weight gain from 7 to 14 days of age. The inhibition of gastric acid secretion decreased curd formation in the abomasum as well as daily weight gain compared to non-treated control calves. This suggested that curd formation in the abomasum is important for the weight gain of newborn calves.

Effect of astaxanthin in combination with alpha-tocopherol or ascorbic acid against oxidative damage in diabetic ODS rats.

The present study was performed to investigate the effect of astaxanthin in combination with other antioxidants against oxidative damage in streptozotocin (STZ)-induced diabetic Osteogenic Disorder Shionogi (ODS) rats. Diabetic-ODS rats were divided into five groups: control, astaxanthin, ascorbic acid, alpha-tocopherol, and tocotrienol. Each of the four experimental groups was administered a diet containing astaxanthin (0.1 g/kg), in combination with ascorbic acid (3.0 g/kg), alpha-tocopherol (0.1 g/kg), or tocotrienol (0.1 g/kg) for 20 wk. The effects of astaxanthin with other antioxidants on lipid peroxidation, urinary 8-hydroxy-2-deoxyguanosine (8-OHdG) excretion, serum creatinine (Cr) level, creatinine clearance (Ccr), and urinary protein content were assessed. The serum lipid peroxide levels and chemiluminescent (CL) intensity in the liver of the alpha-tocopherol and tocotrienol groups were significantly reduced in comparison to that of the control group. In the alpha-tocopherol group, urinary 8-OHdG excretion, serum Cr level, Ccr, urinary albumin excretion, and urinary protein concentration were significantly decreased as compared with those in the control group. Additionally, the CL intensity in the kidney of the alpha-tocopherol group was significantly lower, but that of the ascorbic acid group was significantly higher than that in the control group. These results indicate that dietary astaxanthin in combination with alpha-tocopherol has an inhibitory effect on oxidative stress. On the other hand, our study suggests that excessive ascorbic acid intake increases lipid peroxidation in diabetic rats.