Design and validation of a real-time PCR technique for assessing the level of inclusion of fungus- and yeast-based additives in feeds.

A reliable method for quantification of non-viable microbe-based nutritional and zootechnical additives introduced into feed is essential in order to ensure regulatory compliance, feed safety and product authenticity in industrial applications. In the present work, we developed a novel real-time quantitative polymerase chain reaction (qPCR) -based analysis protocol for monitoring two microbial additives in feed matrices. To evaluate the applicability of the method, pelleted wheat- and maize-based broiler chicken diets containing a non-viable phytase-producing strain of Aspergillus niger produced in solid state fermentation (150 or 300 g/t) and a non-viable selenium-enriched Saccharomyces cerevisiae (100 or 200 g/t) as model feed ingredients, were manufactured and subjected to analysis. Power analysis of the qPCR results indicated that 2 to 6 replicate feed samples were required to distinguish the product doses applied, which confirms that the microbial DNA was efficiently recovered and that potential PCR inhibitors present in the feed material were successfully removed in DNA extraction. The analysis concept described here was shown to be an accurate and sensitive tool for monitoring the inclusion levels of non-viable, unculturable microbial supplements in animal diets.

Vitamin E supplementation fails to impact measures of oxidative stress or the anaemia of feline chronic kidney disease: a randomised, double-blinded placebo control study.

This study was designed to test the hypothesis that supplementation with vitamin E, an antioxidant, in cats with chronic kidney disease (CKD), would reduce oxidative stress and its impact on RBC membrane fragility, resulting in these cats maintaining a greater packed cell volume (PCV) compared with CKD cats not receiving supplementation. Thirty-six cats with CKD were randomly assigned to receive either daily vitamin E or a placebo for 3 months in a double-blinded study design. History and physical examination, blood pressure, complete blood count (CBC), PCV, biochemical profile and urinalysis (UA) were determined. Parameters of oxidative stress and osmotic fragility were measured. Cats were administered vitamin E or placebo once daily for 3 months. Cats were then reassessed and the diagnostics were repeated. Twenty-four cats completed the study, 11 in the vitamin E group and 13 in the placebo group. There were no significant differences between the two groups at the start, or upon completion of the study with regard to biochemical parameters, oxidative stress, erythrocyte osmotic fragility or PCV. None of these parameters changed significantly in either group over the treatment period. Daily supplementation with 30 IU of vitamin E did not affect the measures of oxidative stress or the anaemia seen in cats with CKD.