Abstract
Bovine tuberculosis (bTB) is an infectious disease with significant impact on animal health, public health and international trade. Standard bTB screening in live cattle consists in injecting tuberculin and measuring the swelling at the place of injection few days later. This procedure is expensive, time-consuming, logistically challenging, and is not conclusive before performing confirmatory tests and additional analysis. The analysis of the volatile organic compounds (VOCs) emitted by non-invasive biological samples can provide an alternative diagnostic approach suitable for bTB screening. In the present study, we analyzed VOC samples emitted through the breath, feces and skin of 18 cows diagnosed with bTB from three farms from Romania, as well as of 27 negative cows for bTB from the same farms. Analytical studies employing gas chromatography coupled to mass spectrometry revealed 80 VOCs emitted through the breath, 200 VOCs released by feces, and 80 VOCs emitted through the skin. Statistical analysis of these compounds allowed the identification of 3 tentative breath VOC biomarkers (acetone; 4-methyldecane; D-limonene), 9 tentative feces VOC biomarkers (toluene; [(1,1-dimethylethyl)thio]acetic acid; alpha-thujene; camphene; phenol; o-cymene; 3-(1,1-dimethylethyl)-2,2,4,4-tetramethyl-3-pentanol; 2,5-dimethylhexane-2,5-dihydroperoxide; 2,4-di-tert-butylphenol), and 3 tentative skin VOC biomarkers (ammonia; 1-methoxy-2-propanol; toluene). The possible pathway of these volatile biomarkers is discussed.
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Data availability
The datasets generated and/or analyzed during the current study are available from the corresponding author on reasonable request.
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Acknowledgements
We wish to express special thanks to Prof. Danut Turcu, Spiru Haret University, Romania, for his involvement in identifying bovine tuberculosis outbreaks in Romania and samples collection, and to Dr. Pauline Nol, United States Department of Agriculture, USA, for assessment in samples collection and kindly supply of the samples collection material.
Funding
This work was supported by the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie Grant No. 777832, and COMBIVET ERA Chair Grant No. 857418.
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RI contributed to the study conception and design; VES and VA contributed to animal characterization; MB, ALJM and KJBS contributed to chromatographic analysis; FI contributed to statistical analysis; RI, TGW, RS and JBL contributed to biomarkers investigation; RI wrote the main manuscript text; ALJM prepared the figures. All authors reviewed the manuscript.
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The study was approved by the Research Ethics Committee of the Faculty of Veterinary Medicine from Spiru Haret University, Romania (Registry no. 85/13.03.2018). Samples were collected non-invasively from animals as described in the Material and Methods section. No additional interventions were applied to the animals for the scope of this study.
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Brebu, M., Simion, V.E., Andronie, V. et al. Putative volatile biomarkers of bovine tuberculosis infection in breath, skin and feces of cattle. Mol Cell Biochem 478, 2473–2480 (2023). https://doi.org/10.1007/s11010-023-04676-5
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DOI: https://doi.org/10.1007/s11010-023-04676-5