Serum responsiveness to recombinant bovine somatotropin in buffalo: a three‐month lactation study using an acid‐stripping ELISA for screening

Abstract

The misuse of recombinant bovine somatotropin (rbST) to increase milk yield involves buffalo not just cows. Screening methods to identify rbST-treated cattle have already been proposed. However, there have been no studies on prolonged periods with a high number of animals. In this study, we developed a new enzyme-linked immunosorbent assay (ELISA) to measure the serum responsiveness towards rbST, based on an acid-stripping procedure and relatively simple integral calculation dilution curves. We also applied the analysis to 640 serum and 96 milk samples collected from 16 buffalo treated with rbST and 16 controls, over a period of approximately three months. Its suitability as a screening method, in compliance with EU law, was also assessed. A bi-factorial approach was also evaluated, including the measurement of insulin-like growth factor 1 concentration in serum. Results showed that our ELISA could be used on its own for screening purposes, without the need to assess other biomarkers. Copyright © 2016 John Wiley & Sons, Ltd.

Tracing recombinant bovine somatotropin ab(use) through transcriptomics: the potential of bovine somatic cells in a multi-dose longitudinal study

“…Therefore, it is also of great importance to develop indirect methods that allow detection of rbST administration in cattle. In this sense, some research works have developed ELISA methods to detect rbST antibodies in serum and milk 19,20 . Other studies have developed methods based in the determination of multiple protein biomarkers to detect the use of rbST in dairy cattle 5,21 .…”

Tracing (r)bST in cattle: Liquid-based options for extraction and separation

“…Recently, Castigliego et al described a very simple and straightforward method to isolate rbST (somavubove) from slow-release preparations, i.e. Boostin® [28,33] . The reported method consists of two successive carbonate buffer (pH 9.6) additions, vortexing and cool centrifugation, in order to separate the grease layer from the protein solution.…”

“…This procedure has been used to measure the responsiveness of the animal to rbST treatments, using preferably milk and serum as target matrices. A new ELISA based on an acid-stripping procedure was developed very recently to measure buffalo serum and milk responsiveness towards rbST [33] . Antibody production for these assays is a complicated task due to the high degree of homology of rbST, particularly sometribove and somavubove, and bST.…”

Microsphere Peptide-Based Immunoassay for the Detection of Recombinant Bovine Somatotropin in Injection Preparations

“…To distinguish between farms suspected and not-suspected of rbST misuse, a straightforward immunochemical method measuring rbST induced-antibodies can be performed as an initial screening procedure. To measure the rbST-induced antibody expression, the full rbST protein is attached to a variety of surfaces, e.g., microspheres for suspension arrays [14,15], 96-well plates in ELISA [16,17] and glass chips in microarrays [18], enabling detection by binding to the immobilized rbST [19]. As these assays rely on the banned full rbST protein, they suffer from limited accessibility and applicability.…”