Putative volatile biomarkers of bovine tuberculosis infection in breath, skin and feces of cattle.

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.

Non-Invasive Method to Detect Infection with Mycobacterium tuberculosis Complex in Wild Boar by Measurement of Volatile Organic Compounds Obtained from Feces with an Electronic Nose System.

More effective methods to detect bovine tuberculosis, caused by Mycobacterium bovis, in wildlife, is of paramount importance for preventing disease spread to other wild animals, livestock, and human beings. In this study, we analyzed the volatile organic compounds emitted by fecal samples collected from free-ranging wild boar captured in Doñana National Park, Spain, with an electronic nose system based on organically-functionalized gold nanoparticles. The animals were separated by the age group for performing the analysis. Adult (>24 months) and sub-adult (12-24 months) animals were anesthetized before sample collection, whereas the juvenile (<12 months) animals were manually restrained while collecting the sample. Good accuracy was obtained for the adult and sub-adult classification models: 100% during the training phase and 88.9% during the testing phase for the adult animals, and 100% during both the training and testing phase for the sub-adult animals, respectively. The results obtained could be important for the further development of a non-invasive and less expensive detection method of bovine tuberculosis in wildlife populations.

Assessment of Electronic Sensing Techniques for the Rapid Identification of Alveolar Echinococcosis through Exhaled Breath Analysis.

Here we present a proof-of-concept study showing the potential of a chemical gas sensors system to identify the patients with alveolar echinococcosis disease through exhaled breath analysis. The sensors system employed comprised an array of three commercial gas sensors and a custom gas sensor based on WO3 nanowires doped with gold nanoparticles, optimized for the measurement of common breath volatile organic compounds. The measurement setup was designed for the concomitant measurement of both sensors DC resistance and AC fluctuations during breath samples exposure. Discriminant Function Analysis classification models were built with features extracted from sensors responses, and the discrimination of alveolar echinococcosis was estimated through bootstrap validation. The commercial sensor that detects gases such as alkane derivatives and ethanol, associated with lipid peroxidation and intestinal gut flora, provided the best classification (63.4% success rate, 66.3% sensitivity and 54.6% specificity) when sensors' responses were individually analyzed, while the model built with the AC features extracted from the responses of the cross-reactive sensors array yielded 90.2% classification success rate, 93.6% sensitivity and 79.4% specificity. This result paves the way for the development of a noninvasive, easy to use, fast and inexpensive diagnostic test for alveolar echinococcosis diagnosis at an early stage, when curative treatment can be applied to the patients.

Challenges during the realization of an international research project on leishmaniasis in Colombia.

Leishmaniasis is an infectious disease that belongs to the top 10 neglected tropical diseases. It mainly affects the poor population from tropical and subtropical areas of the World, which lacks sufficient resources and means to fight against this disease. With this in mind, the European Commission has funded an international collaborative research project in which are participating various institutions from South America, North Africa and Europe. The main objective of this project is the development of a fast, less expensive, non-invasive and easy to use alternative method for leishmaniasis diagnosis in dogs, one of the main reservoirs of leishmaniasis spread to humans. In this perspective article, we present our personal insight and opinion regarding the challenges of realizing a joint international research project on leishmaniasis in Colombia, a country where leishmaniasis is endemic, as well as regarding the involvement of the Public Health institutions and the local population from this country.

Review on Volatolomic Studies as a Frontier Approach in Animal Research.

This paper presents a comprehensive review of the research studies in volatolomics performed on animals so far. At first, the procedures proposed for the collection, preconcentration, and storing of the volatile organic compounds emitted by various biological samples of different animals are presented and discussed. Next, the results obtained in the analysis of the collected volatile samples with analytical equipment are shown. The possible volatile biomarkers identified for various diseases are highlighted for different types of diseases, animal species, and biological samples analyzed. The chemical classes of these compounds, as well as the biomarkers found in a higher number of animal diseases, are indicated, and their possible origin is analyzed. The studies that dealt with the diagnosis of various diseases from sample measurement with electronic nose systems are also presented and discussed. The paper ends with a final remark regarding the necessity of optimization and standardization of sample collection and analysis procedures for obtaining meaningful results.

Exhaled breath analysis for gastric cancer diagnosis in Colombian patients.

We present here the first study that directly correlates gastric cancer (GC) with specific biomarkers in the exhaled breath composition on a South American population, which registers one of the highest global incidence rates of gastric affections. Moreover, we demonstrate a novel solid state sensor that predicts correct GC diagnosis with 97% accuracy. Alveolar breath samples of 30 volunteers (patients diagnosed with gastric cancer and a controls group formed of patients diagnosed with other gastric diseases) were collected and analyzed by gas-chromatography/mass-spectrometry (GC-MS) and with an innovative chemical gas sensor based on gold nanoparticles (AuNP) functionalized with octadecylamine ligands. Our GC-MS analyses identified 6 volatile organic compounds that showed statistically significant differences between the cancer patients and the controls group. These compounds were different from those identified in previous studied performed on other populations with high incidence rates of this malady, such as China (representative for Eastern Asia region) and Latvia (representative for Baltic States), attributable to lifestyle, alimentation and genetics differences. A classification model based on principal component analysis of our sensor data responses to the breath samples yielded 97% accuracy, 100% sensitivity and 93% specificity. Our results suggest a new and non-intrusive methodology for early diagnosis of gastric cancer that may be deployed in regions lacking well-developed health care systems as a prediagnosis test for selecting the patients that should undergo deeper investigations (e.g., endoscopy and biopsy).