Potential of Treatment-Specific Protein Biomarker Profiles for Detection of Hormone Abuse in Cattle

Ludwig, Susann K. J.; Smits, Nathalie G. E.; Cannizzo, Francesca Tiziana; Nielen, Michel W. F.

doi:10.1021/jf4004972

Cited by 16 publications

(18 citation statements)

References 45 publications

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“…The microarray chip was subsequently incubated with milk sample extract, secondary antibodies and Quantum Dots, whereupon the microarray spots were fluorescently labelled depending on the amount of biomarker present in the sample extract. The microarray chip was imaged following insertion into the low-cost smartphone attachment housing having UV LEDs for fluorescence excitation, an external lens for an increased field of view, and an optical emission light filter ( Fig 1A and 1B ) [ 24 ]. With our custom-developed Android application “Spot-An-Array”, the user can decide whether an image should be taken or whether a previously taken image shall be analyzed ( Fig 2 ).…”

Section: Resultsmentioning

confidence: 99%

Calling Biomarkers in Milk Using a Protein Microarray on Your Smartphone

et al. 2015

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Here we present the concept of a protein microarray-based fluorescence immunoassay for multiple biomarker detection in milk extracts by an ordinary smartphone. A multiplex immunoassay was designed on a microarray chip, having built-in positive and negative quality controls. After the immunoassay procedure, the 48 microspots were labelled with Quantum Dots (QD) depending on the protein biomarker levels in the sample. QD-fluorescence was subsequently detected by the smartphone camera under UV light excitation from LEDs embedded in a simple 3D-printed opto-mechanical smartphone attachment. The somewhat aberrant images obtained under such conditions, were corrected by newly developed Android-based software on the same smartphone, and protein biomarker profiles were calculated. The indirect detection of recombinant bovine somatotropin (rbST) in milk extracts based on altered biomarker profile of anti-rbST antibodies was selected as a real-life challenge. RbST-treated and untreated cows clearly showed reproducible treatment-dependent biomarker profiles in milk, in excellent agreement with results from a flow cytometer reference method. In a pilot experiment, anti-rbST antibody detection was multiplexed with the detection of another rbST-dependent biomarker, insulin-like growth factor 1 (IGF-1). Milk extract IGF-1 levels were found to be increased after rbST treatment and correlated with the results obtained from the reference method. These data clearly demonstrate the potential of the portable protein microarray concept towards simultaneous detection of multiple biomarkers. We envisage broad application of this ‘protein microarray on a smartphone’-concept for on-site testing, e.g., in food safety, environment and health monitoring.

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Section: Resultsmentioning

confidence: 99%

Calling Biomarkers in Milk Using a Protein Microarray on Your Smartphone

et al. 2015

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“…For many years several research groups have attempted to identify molecular markers useful to detect different anabolic treatments in cattle [27][28][29][30][31] . The usefulness of histological analyses on thymus morphology as a screening tool to identify illegal DEX treatments in cattle was already established [6][7][8][9][10] .…”

Section: Discussionmentioning

confidence: 99%

Morphological Examination and Transcriptomic Profiling To Identify Prednisolone Treatment in Beef Cattle

Cannizzo

Pegolo

Pregel

et al. 2016

J. Agric. Food Chem.

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in skeletal muscle and testis sampled at slaughtering. In the epididymis, spermatozoa number decreased in PDN treated animals, and in some cases they were totally absent. Correspondently, in testis of treated animals, down-regulation for serine/threonine kinase 11 (STK11) gene expression was detected (p<0.01). DNA microarray analysis revealed a total of 133 differentially expressed genes in skeletal muscle and testis, and 907 and 1,416 in PBMCs after 33 days of treatment and at slaughtering, respectively. Histological investigations on epididymal content could represent a promising marker for PDN treatment in beef cattle, and could be used as a screening method in order to identify animals worthy of further investigation with official methods. Moreover, the clear transcriptomic signature of PDN treatment evidenced in PBMCs supported for the possibility of using this matrix to monitor the illicit treatment in vivo during ranching.

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“…The use of biomarkers is becoming more and more popular, both in medicine, for indication or diagnosis on possible diseases, and in legal analysis addressed to reveal doping abuse in sports and illegal or undeclared pharmacological treatment in livestock [1][2][3][4][5]. This is due to the fact that biomarkers may be often easily quantified or create characteristic patterns that may be profiled in a more simple and cheaper way than other direct methods, such as the research and quantification of residues in biological materials.…”

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

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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.

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Potential of Treatment-Specific Protein Biomarker Profiles for Detection of Hormone Abuse in Cattle

Cited by 16 publications

References 45 publications

Calling Biomarkers in Milk Using a Protein Microarray on Your Smartphone

Calling Biomarkers in Milk Using a Protein Microarray on Your Smartphone

Morphological Examination and Transcriptomic Profiling To Identify Prednisolone Treatment in Beef Cattle

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

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