Recording the Presence of Peanibacillus larvae larvae Colonies on MYPGP Substrates Using a Multi-Sensor Array Based on Solid-State Gas Sensors

American foulbrood is a dangerous disease of bee broods found worldwide, caused by the Paenibacillus larvae larvae L. bacterium. In an experiment, the possibility of detecting colonies of this bacterium on MYPGP substrates (which contains yeast extract, Mueller-Hinton broth, glucose, K2HPO4, sodium pyruvate, and agar) was tested using a prototype of a multi-sensor recorder of the MCA-8 sensor signal with a matrix of six semiconductors: TGS 823, TGS 826, TGS 832, TGS 2600, TGS 2602, and TGS 2603 from Figaro. Two twin prototypes of the MCA-8 measurement device, M1 and M2, were used in the study. Each prototype was attached to two laboratory test chambers: a wooden one and a polystyrene one. For the experiment, the strain used was P. l. larvae ATCC 9545, ERIC I. On MYPGP medium, often used for laboratory diagnosis of American foulbrood, this bacterium produces small, transparent, smooth, and shiny colonies. Gas samples from over culture media of one- and two-day-old foulbrood P. l. larvae (with no colonies visible to the naked eye) and from over culture media older than 2 days (with visible bacterial colonies) were examined. In addition, the air from empty chambers was tested. The measurement time was 20 min, including a 10-min testing exposure phase and a 10-min sensor regeneration phase. The results were analyzed in two variants: without baseline correction and with baseline correction. We tested 14 classifiers and found that a prototype of a multi-sensor recorder of the MCA-8 sensor signal was capable of detecting colonies of P. l. larvae on MYPGP substrate with a 97% efficiency and could distinguish between MYPGP substrates with 1–2 days of culture, and substrates with older cultures. The efficacy of copies of the prototypes M1 and M2 was shown to differ slightly. The weighted method with Canberra metrics (Canberra.811) and kNN with Canberra and Manhattan metrics (Canberra. 1nn and manhattan.1nn) proved to be the most effective classifiers.

[1]  Beata Bak,et al.  Diagnosis of Varroosis Based on Bee Brood Samples Testing with Use of Semiconductor Gas Sensors , 2020, Sensors.

[2]  E. Brzuszkiewicz,et al.  How to Kill the Honey Bee Larva: Genomic Potential and Virulence Mechanisms of Paenibacillus larvae , 2014, PloS one.

[3]  H. T. Nagle,et al.  Handbook of Machine Olfaction , 2002 .

[4]  J. Berdagué,et al.  Rapid discrimination of Micrococcaceae species using semiconductor gas sensors , 1995 .

[5]  László Györfi,et al.  A Probabilistic Theory of Pattern Recognition , 1996, Stochastic Modelling and Applied Probability.

[6]  W. Lubitz,et al.  Classification and identification of bacteria: current approaches to an old problem. Overview of methods used in bacterial systematics. , 1996, Journal of biotechnology.

[7]  Ananya Dey,et al.  Semiconductor metal oxide gas sensors: A review , 2018 .

[8]  E. Genersch,et al.  Biology of Paenibacillus larvae, a deadly pathogen of honey bee larvae , 2016, Applied Microbiology and Biotechnology.

[9]  K. Triyana,et al.  Gas Sensor Array System Properties for Detecting Bacterial Biofilms , 2019, Journal of medical signals and sensors.

[10]  M. Rohde,et al.  Discovery of Paenibacillus larvae ERIC V: Phenotypic and genomic comparison to genotypes ERIC I-IV reveal different inventories of virulence factors which correlate with epidemiological prevalences of American Foulbrood. , 2020, International journal of medical microbiology : IJMM.

[11]  Maryam Siadat,et al.  Orthogonal Signal Correction to Improve Stability Regression Model in Gas Sensor Systems , 2017, J. Sensors.

[12]  J. Haugen,et al.  A calibration method for handling the temporal drift of solid state gas-sensors , 2000 .

[13]  Julian W. Gardner,et al.  A brief history of electronic noses , 1994 .

[14]  J. Gardner,et al.  Biomedical Engineering Online Open Access Bacteria Classification Using Cyranose 320 Electronic Nose , 2022 .

[15]  E. Domashevskaya,et al.  APPLICATION OF SEMICONDUCTOR GAS SENSORS FOR MEDICAL DIAGNOSTICS , 1999 .

[16]  L. Francioso,et al.  Chemiresistor gas sensors using semiconductor metal oxides , 2014 .

[17]  A. Lindström Distribution of Paenibacillus larvae Spores Among Adult Honey Bees (Apis mellifera) and the Relationship with Clinical Symptoms of American Foulbrood , 2008, Microbial Ecology.

[18]  E. Genersch,et al.  Rapid identification of differentially virulent genotypes of Paenibacillus larvae, the causative organism of American foulbrood of honey bees, by whole cell MALDI-TOF mass spectrometry. , 2014, Veterinary microbiology.

[19]  I. Fries,et al.  Reclassification of Paenibacillus larvae subsp. pulvifaciens and Paenibacillus larvae subsp. larvae as Paenibacillus larvae without subspecies differentiation. , 2006, International journal of systematic and evolutionary microbiology.

[20]  D. Shearer,et al.  Volatile Acids from Honeybee Larvae Infected with Bacillus Larvae and from a Culture of the Organism , 1981 .

[21]  David Smith,et al.  Detection of volatile compounds emitted by Pseudomonas aeruginosa using selected ion flow tube mass spectrometry , 2005, Pediatric pulmonology.

[22]  Birgit Piechulla,et al.  Bacterial volatiles and their action potential , 2009, Applied Microbiology and Biotechnology.

[23]  K. Triyana,et al.  Lab-Made Electronic Nose for Fast Detection of Listeria monocytogenes and Bacillus cereus , 2020, Veterinary sciences.

[24]  J. Ellis,et al.  The worldwide health status of honey bees , 2005 .

[25]  Khalil Arshak,et al.  A review of gas sensors employed in electronic nose applications , 2004 .

[26]  Monika Maciejewska,et al.  Gas Sensor Array and Classifiers as a Means of Varroosis Detection , 2019, Sensors.

[27]  M. Siadat,et al.  Reduction of drift impact in gas sensor response to improve quantitative odor analysis , 2017, 2017 IEEE International Conference on Industrial Technology (ICIT).

[28]  A. D. Wilson,et al.  Applications of Electronic-Nose Technologies for Noninvasive Early Detection of Plant, Animal and Human Diseases , 2018, Chemosensors.

[29]  Changsheng Xie,et al.  Metal-oxide-semiconductor based gas sensors: screening, preparation, and integration. , 2017, Physical chemistry chemical physics : PCCP.

[30]  Lech Polkowski,et al.  Granular Computing in Decision Approximation - An Application of Rough Mereology , 2015, Intelligent Systems Reference Library.

[31]  Roberto Paolesse,et al.  Identification of a Large Pool of Microorganisms with an Array of Porphyrin Based Gas Sensors , 2016, Sensors.

[32]  Jay D. Evans,et al.  Standard methods for American foulbrood research , 2013 .

[33]  Keshun Yu,et al.  Comparison of long‐chain alcohols and other volatile compounds emitted from food‐borne and related Gram positive and Gram negative bacteria , 2002, Journal of basic microbiology.

[34]  P. Hauptmann Sensors: A Comprehensive Survey , 1996 .

[35]  J. Aslanzadeh,et al.  Biochemical Profile-Based Microbial Identification Systems , 2013 .

[36]  P. John Clarkson,et al.  Early detection of diseases in tomato crops: An Electronic Nose and intelligent systems approach , 2010, The 2010 International Joint Conference on Neural Networks (IJCNN).

[37]  Richard O. Duda,et al.  Pattern classification and scene analysis , 1974, A Wiley-Interscience publication.

[38]  E. Genersch American Foulbrood in honeybees and its causative agent, Paenibacillus larvae. , 2010, Journal of invertebrate pathology.

[39]  F. Margara,et al.  Attraction of Mexican fruit flies (Diptera: Tephritidae) to bacteria: effects of culturing medium on odour volatiles , 2009 .

[40]  H. Troy Nagle,et al.  Handbook of Machine Olfaction: Electronic Nose Technology , 2003 .

[41]  J. Shieh,et al.  Development of an E-nose system using machine learning methods to predict ventilator-associated pneumonia , 2020, Microsystem Technologies.

[42]  Myeong-lyeol Lee,et al.  Volatile disease markers of American foulbrood-infected larvae in Apis mellifera. , 2020, Journal of insect physiology.

[43]  Jeroen S. Dickschat,et al.  The scent of bacteria: headspace analysis for the discovery of natural products. , 2012, Journal of natural products.