Monitoring milk for antibodies against recombinant bovine somatotropin using a microsphere immunoassay-based biomarker approach

Ludwig, Susann K. J.; Smits, Nathalie G. E.; Bremer, M.G.E.G.; Nielen, Michel W. F.

doi:10.1016/j.foodcont.2011.12.011

Cited by 18 publications

(21 citation statements)

References 13 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Since blood sampling in routine veterinary monitoring programmes might not be justified in some countries, we suggest a tiered approach according to Commission Decision 2002/657/EC. This would consist of three steps: First, a fast screening for anti-rbST-antibodies in tank milk using our previously described milk FCIA [29]. Second, in case of suspicious findings, a more detailed individual bovine serum biomarker profiling using the IA, AO or IAO FCIA presented here will provide additional evidence, since they are based on more biomarkers and data for individual cows.…”

Section: Discussionmentioning

confidence: 99%

“…A generic sample pre-treatment procedure which was crucial for removing non-specific interferences and making the candidate biomarkers accessible for detection was described previously [4], [28], [29], is depicted in Figure 1 and described in-depth in Materials S1.…”

Section: Methodsmentioning

confidence: 99%

“…Covalent coupling of 100 µg mL −1 Monsanto rbST standard, 100 µg mL −1 IGF-1 and 10 µg mL −1 IGFBP2 to seroMAP microspheres (sets 050, 025, 078 respectively) was described before [4], [28], [29]. Coupling 75 µg mL −1 osteocalcin to microspheres (set 084) was done following the same procedure.…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Multiple Protein Biomarker Assessment for Recombinant Bovine Somatotropin (rbST) Abuse in Cattle

Ludwig¹,

Smits²,

Veer³

et al. 2012

PLoS ONE

View full text Add to dashboard Cite

Biomarker profiling, as a rapid screening approach for detection of hormone abuse, requires well selected candidate biomarkers and a thorough in vivo biomarker evaluation as previously done for detection of growth hormone doping in athletes. The bovine equivalent of growth hormone, called recombinant bovine somatotropin (rbST) is (il)legally administered to enhance milk production in dairy cows. In this study, first a generic sample pre-treatment and 4-plex flow cytometric immunoassay (FCIA) were developed for simultaneous measurement of four candidate biomarkers selected from literature: insulin-like growth factor 1 (IGF-1), its binding protein 2 (IGFBP2), osteocalcin and endogenously produced antibodies against rbST. Next, bovine serum samples from two extensive controlled rbST animal treatment studies were used for in vivo validation and biomarker evaluation. Finally, advanced statistic tools were tested for the assessment of biomarker combination quality aiming to correctly identify rbST-treated animals. The statistical prediction tool k-nearest neighbours using a combination of the biomarkers osteocalcin and endogenously produced antibodies against rbST proved to be very reliable and correctly predicted 95% of the treated samples starting from the second rbST injection until the end of the treatment period and even thereafter. With the same biomarker combination, only 12% of untreated animals appeared false-positive. This reliability meets the requirements of Commission Decision 2002/657/EC for screening methods in veterinary control. From the results of this multidisciplinary study, it is concluded that the osteocalcin – anti-rbST-antibodies combination represent fit-for-purpose biomarkers for screening of rbST abuse in dairy cattle and can be reliably measured in both the developed 4-plex FCIA as well as in a cost-effective 2-plex microsphere-based binding assay. This screening method can be incorporated in routine veterinary monitoring programmes: in the European Union for detection of rbST abuse and in the control of rbST-free dairy farms in the United States of America and other countries.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Multiple Protein Biomarker Assessment for Recombinant Bovine Somatotropin (rbST) Abuse in Cattle

Ludwig¹,

Smits²,

Veer³

et al. 2012

PLoS ONE

View full text Add to dashboard Cite

show abstract

“…These liquid-based systems have demonstrated its applicability also to monitor somatotropin abuse in cattle. A multiplex flow cytometric immunoassay capable of discriminating between milk from rbST-treated and untreated animals was developed by Ludwig et al in 2012 [37] . The method was based on the detection of anti-rbST antibodies in raw milk.…”

Section: Other Liquid Applications and Indirect Determinations Of Rbstmentioning

confidence: 99%

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

Regal

Lamas

Fente

et al. 2017

Journal of Liquid Chromatography & Related Technologies

View full text Add to dashboard Cite

Growth hormone (GH) or somatotropin (ST) is a species-specific polypeptide hormone produced in the pituitary gland of vertebrates. When administered exogenously to dairy cattle it has galactopoietic effects and is capable of increasing the milk yield. The beginning of the commercial production of recombinant variants of bovine somatotropin (rbST), by the end of the twentieth century, had a major economic impact in the dairy industry. Recombinant bST enabled large scale applications in farms, enhancing significantly milk production. While it is banned in the European Union (EU), several countries permit the trade and use of recombinant somatotropins in animal husbandry. Also, rbST-free dairy products can be frequently found in the market of those countries, even though these labels are not actually verified in a laboratory. In this context, effective analytical methods are needed for residue control to avoid an illegal use of rbST but also to prevent fraudulent labelling in some cases. The present review includes studies published in the last five years (from 2012 to 2017) to monitor rbST in bovine animals, using liquid-based applications. It is then intended to serve as a practical guide to help 2 those laboratories interested in developing analytical methods to detect rbST use and abuse.

show abstract

“…Beyond some confirmation methods based on mass spectrometry [8][9][10], which require specialized personnel and expensive instrumentations, efforts have been made to develop cheaper and more widely applicable tests. In this direction, different approaches have been proposed, from the research and quantification of molecules recognizing the somatotropin in serum [11][12][13], through the combined research of a number of different molecules, the concentration of which may vary in serum responding to the rbST treatment cows [14][15][16][17][18], unto the quantification of the alteration of gene expression in different tissues [19,20].…”

Section: Introductionmentioning

confidence: 99%

Selection of Biomarkers from Differentially Expressed Genes in Leukocytes of Buffalos Treated with Recombinant Bovine Somatotropin: The Importance of Sample Size for Reliable Discriminating Systems

et al. 2016

View full text Add to dashboard Cite

The research on biomarkers to detect livestock treated with recombinant bovine somatotropin (rbST) is still an open issue. In fact, beyond undertaking confirmation methods, there is the need to develop simple and inexpensive screening tests. In this direction, some proposals have been forwarded, mostly involving the measurement of circulating molecules, whereas the possibility of using biomarkers related to gene expression is a field under investigation. The present study was carried out on sixteen buffalos, eight of which treated with rbST. Blood samples were collected six times during the treatment to investigate on the presence of differentially expressed genes in leukocytes. Analysis with the microarray technique was performed on two sampling moments, in order to obtain a first selection of genes. Further analysis was carried out by real time RT-PCR, in order to create a discriminating linear system. A study on the variation of the error related to the number of samples included in statistics was also performed. Results showed that, including an increasing number of samples to build the discriminating algorithm, the-error grows and tends to stabilize on 6.5%. This study clearly shows the paramount importance of including a proper number of samples to obtain reliable algorithms.

show abstract

Monitoring milk for antibodies against recombinant bovine somatotropin using a microsphere immunoassay-based biomarker approach

Cited by 18 publications

References 13 publications

Multiple Protein Biomarker Assessment for Recombinant Bovine Somatotropin (rbST) Abuse in Cattle

Multiple Protein Biomarker Assessment for Recombinant Bovine Somatotropin (rbST) Abuse in Cattle

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

Selection of Biomarkers from Differentially Expressed Genes in Leukocytes of Buffalos Treated with Recombinant Bovine Somatotropin: The Importance of Sample Size for Reliable Discriminating Systems

Contact Info

Product

Resources

About