{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,7]],"date-time":"2026-01-07T08:10:37Z","timestamp":1767773437123,"version":"build-2065373602"},"reference-count":35,"publisher":"MDPI AG","issue":"15","license":[{"start":{"date-parts":[[2020,7,26]],"date-time":"2020-07-26T00:00:00Z","timestamp":1595721600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Sensors"],"abstract":"<jats:p>Monitoring the spore life cycle is one of the main issues in several fields including environmental control, sustainable ecosystems, food security, and healthcare systems. In this framework, the study of the living organism resistance to extreme conditions like those mimicking space environments is particularly interesting. The assessment of the local change of the pH level can be extremely useful for this purpose. An optical physiometer method based on the Raman response of the graphene, which is able to locally sense pH of a fluid on a micrometric scale, has been recently proposed. Due to the presence of    \u03c0   -bonds at the surface, the electronic doping of graphene is determined by the external conditions and can be electrochemically controlled or altered by the contact with an acid or alkaline fluid. The doping level affects the vibrational energies of the graphene that can be monitored by conventional Raman spectroscopy. In addition, Surface-Enhanced Raman Spectroscopy (SERS) can give direct information on the biochemical changes occurring in spore components. In this work, we propose the joint use of Graphene-Based Raman Spectroscopy (GbRS) and SERS for the monitoring of the response of spores to exposure to low temperatures down to 100 K. The spores of the thermophilic bacterium Parageobacillus thermantarcticus isolated from an active volcano of Antarctica (Mt. Melbourne) were investigated. These spores are particularly resistant to several stressing stimuli and able to adapt to extreme conditions like low temperatures, UV irradiation, and    \u03b3   -rays exposure. The results obtained showed that the joint use of GbRS and SERS represents a valuable tool for monitoring the physio-chemical response of bacterial spores upon exposure to stressing stimuli.<\/jats:p>","DOI":"10.3390\/s20154150","type":"journal-article","created":{"date-parts":[[2020,7,27]],"date-time":"2020-07-27T04:39:50Z","timestamp":1595824790000},"page":"4150","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Graphene-Based and Surface-Enhanced Raman Spectroscopy for Monitoring the Physio-Chemical Response of Thermophilic Bacterial Spores to Low Temperatures Exposure"],"prefix":"10.3390","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6014-0588","authenticated-orcid":false,"given":"Carlo","family":"Camerlingo","sequence":"first","affiliation":[{"name":"CNR-SPIN, Institute for Superconductivity, Innovative Materials and Devices, 80078 Pozzuoli, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Giuseppe","family":"Di Meo","sequence":"additional","affiliation":[{"name":"CNR-ICB, Institute of Biomolecular Chemistry, 80078 Pozzuoli, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2743-5904","authenticated-orcid":false,"given":"Maria","family":"Lepore","sequence":"additional","affiliation":[{"name":"Dipartimento di Medicina Sperimentale, Universit\u00e0 della Campania \u201cL. Vanvitelli\u201d, Via S. Maria di Costantinopoli 16, 80138 Napoli, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4791-6468","authenticated-orcid":false,"given":"Mikhail","family":"Lisitskiy","sequence":"additional","affiliation":[{"name":"CNR-SPIN, Institute for Superconductivity, Innovative Materials and Devices, 80078 Pozzuoli, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6465-4765","authenticated-orcid":false,"given":"Annarita","family":"Poli","sequence":"additional","affiliation":[{"name":"CNR-ICB, Institute of Biomolecular Chemistry, 80078 Pozzuoli, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4463-0779","authenticated-orcid":false,"given":"Marianna","family":"Portaccio","sequence":"additional","affiliation":[{"name":"Dipartimento di Medicina Sperimentale, Universit\u00e0 della Campania \u201cL. Vanvitelli\u201d, Via S. Maria di Costantinopoli 16, 80138 Napoli, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7203-9869","authenticated-orcid":false,"given":"Ida","family":"Romano","sequence":"additional","affiliation":[{"name":"CNR-ICB, Institute of Biomolecular Chemistry, 80078 Pozzuoli, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7485-1815","authenticated-orcid":false,"given":"Paola","family":"Di Donato","sequence":"additional","affiliation":[{"name":"CNR-ICB, Institute of Biomolecular Chemistry, 80078 Pozzuoli, Italy"},{"name":"Dipartimento di Scienze e Tecnologie, Universit\u00e0 Parthenope di Napoli, Centro Direzionale Isola C\/4, 80143 Napoli, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,7,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"351","DOI":"10.1007\/s11084-007-9074-5","article-title":"Question 2: Relation of panspermia-hypothesis to astrobiology","volume":"37","author":"Zagorski","year":"2007","journal-title":"Orig. Life Evol. Biosph."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1297","DOI":"10.1128\/JB.01455-13","article-title":"Germination of spores of bacillus species: What we know and do not know","volume":"196","author":"Setlow","year":"2014","journal-title":"J. Bacteriol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"103470","DOI":"10.1016\/j.fm.2020.103470","article-title":"Survival of Salmonella enterica and shifts in the culturable mesophilic aerobic bacterial community as impacted by tomato wash water particulate size and chlorine treatment","volume":"90","author":"Luo","year":"2020","journal-title":"Food Microbiol."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Vidic, J., Chaix, C., Manzano, M., and Heyndrickx, M. (2020). Food sensing: Detection of Bacillus cereus spores in dairy products. Biosensors, 10.","DOI":"10.3390\/bios10030015"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1016\/j.tifs.2019.03.016","article-title":"Heat assisted HPP for inactivation of bacteria, moulds and yeasts spores in foods: Log reductions and mathematical models","volume":"88","author":"Evelyn","year":"2019","journal-title":"Trends Food Sci. Technol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1016\/j.tifs.2016.09.008","article-title":"Processed cheese contamination by spore-forming bacteria: A review of source, routes, fate during processing and control","volume":"57","author":"Oliveira","year":"2016","journal-title":"Trends Food Sci. Technol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"746","DOI":"10.1094\/Phyto-84-746","article-title":"Role of volatile compounds from wounded oranges in induction of germination of Penicillum digitatum Conidia","volume":"84","author":"Eckert","year":"1994","journal-title":"Phytopathology"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"3220","DOI":"10.1128\/AEM.64.9.3220-3224.1998","article-title":"Comparative study of pressure-induced germination of Bacillus subtilis spores at low and high pressure","volume":"64","author":"Wuytack","year":"1998","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_9","first-page":"2","article-title":"Incorporating germination-induction into decontamination strategies for bacterial spores","volume":"124","author":"Kolher","year":"2017","journal-title":"J. Appl. Microbiol."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"S\u00e1nchez-Clemente, R., Ige\u00f1o, M.I., Poblati\u00f3n, A.G., Guijo, M.I., Merch\u00e1n, F., and Blasco, R. (2018). Study of pH changes in media during bacterial growth of several environmental strains. Proceedings, 2.","DOI":"10.3390\/proceedings2201297"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"6865","DOI":"10.1038\/srep06865","article-title":"A graphene-based physiometer array for the analysis of single biological cells","volume":"4","author":"Paulus","year":"2014","journal-title":"Sci. Rep."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"210","DOI":"10.1038\/nnano.2008.67","article-title":"Monitoring dopants by Raman scattering in a electrochemical top-gated graphene transistor","volume":"3","author":"Das","year":"2008","journal-title":"Nat. Nanotechnol."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Camerlingo, C., Verde, A., Manti, L., Meschini, R., Delfino, I., and Lepore, M. (2018). Graphene-based Raman spectroscopy for pH sensing of X-rays exposed and unexposed culture media and cells. Sensors, 18.","DOI":"10.3390\/s18072242"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"931","DOI":"10.1007\/s00792-018-1049-0","article-title":"Resistance and Raman spectroscopy analysis of Parageobacillus thermantarcticus spores after \u03b3-ray exposure","volume":"22","author":"Romano","year":"2018","journal-title":"Extremophiles"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"3683","DOI":"10.1039\/c2an35468c","article-title":"Analysis of the Raman spectra of Ca2+-dipinolinic acid alone and in the bacterial spore core in both aqueous and deydrated environments","volume":"137","author":"Kong","year":"2012","journal-title":"Analyst"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"868","DOI":"10.1366\/000370203322102979","article-title":"Analysis of single bacterial spores by micro-Raman spectroscopy","volume":"57","author":"Esposito","year":"2003","journal-title":"Appl. Spectrosc."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"4681","DOI":"10.1128\/JB.00282-07","article-title":"Levels of Ca2+-dipinolinic acid in individual Bacillus spores determined using microfluidic Raman twizeers","volume":"189","author":"Huang","year":"2007","journal-title":"J. Bacteriol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1366\/000370208783759623","article-title":"Bacillus spores classification via surface-enhanced Raman spectroscopy and principal component analysis","volume":"62","author":"Guicheteau","year":"2007","journal-title":"Appl. Spectrosc."},{"key":"ref_19","unstructured":"Farquharson, S., Shende, C., Gift, A., and Inscore, F. (2020, July 26). Detection of Bacillus Spores by Surface-Enhanced Raman Spectroscopy. Available online: http:\/\/www.intechopen.com\/books\/bioterrorism\/detection-of-bacillus-spores-by-surface-enhanced-raman-spectroscopy."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"4756","DOI":"10.1002\/anie.201205748","article-title":"Surface-Enhanced Raman Spectroscopy: Concepts and chemical applications","volume":"53","year":"2014","journal-title":"Angew. Chem. Int. Ed."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"125901","DOI":"10.1088\/0034-4885\/78\/12\/125901","article-title":"The physics of Martian weather and climate: A review","volume":"78","author":"Read","year":"2015","journal-title":"Rep. Prog. Phys."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Finore, I., Lama, L., Di Donato, P., Romano, I., Tramice, A., Leone, L., Nicolaus, B., and Poli, A. (2019). Parageobacillus thermantarcticus, an Antarctic cell factory: From crop residue valorization by green chemistry to astrobiology studies. Diversity, 11.","DOI":"10.3390\/d11080128"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1007\/s11084-014-9397-y","article-title":"Extremophiles survival to simulated space conditions: An astrobiology model study","volume":"44","author":"Mastascusa","year":"2014","journal-title":"Orig. Life Evol. Biosph."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1007\/s11084-017-9540-7","article-title":"Survival and adaptation of the thermophilic species Geobacillus thermantarcticus in simulated spatial conditions","volume":"48","author":"Romano","year":"2018","journal-title":"Orig. Life Evol. Biosph."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Camerlingo, C., Portaccio, M., Tat\u00e9, R., Lepore, M., and Delfino, I. (2017). Fructose and pectin detection in fruit-based food products by Surface-Enhanced Raman Spectroscopy. Sensors, 17.","DOI":"10.3390\/s17040839"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Camerlingo, C., Lisitskiy, M., Lepore, M., Portaccio, M., Montorio, D., Del Prete, S., and Cennamo, G. (2019). characterization of human tear fluid by means of Surface-enhanced Raman spectroscopy. Sensors, 19.","DOI":"10.3390\/s19051177"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1016\/j.chemolab.2012.03.011","article-title":"Spectral pre-processing for biomedical vibrational spectroscopy and micro-spectroscopic imaging","volume":"117","author":"Lasch","year":"2013","journal-title":"Chemometr. Intell. Lab. Syst."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1140\/epjti\/s40485-015-0018-6","article-title":"Review of multidimensional data processing approaches for Raman and infrared spectroscopy","volume":"2","author":"Gautam","year":"2015","journal-title":"EPJ Tech. Instrum."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"5775","DOI":"10.1128\/AEM.01669-16","article-title":"q Use of Raman spectroscopy and phase-contrast microscopy to characterize cold atmospheric plasma Inactivation of individual bacterial spores","volume":"82","author":"Wang","year":"2016","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"187401","DOI":"10.1103\/PhysRevLett.97.187401","article-title":"Raman spectrum of graphene and graphene layers","volume":"97","author":"Ferrari","year":"2006","journal-title":"Phys. Rev. Lett."},{"key":"ref_31","unstructured":"The value of n was estimated using the relation n = (6817.6 + 8.63\u03bdG \u2212 0.0027\u03bdG2)1013, that approximates the experimental dependence of Ref. [12] for n in the range between \u22124 \u00d7 1012 and \u221225 \u00d7 1012 cm\u22122."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"296","DOI":"10.1016\/j.tim.2013.03.001","article-title":"Mechanisms of endospore inactivation under high pressure","volume":"21","author":"Reineke","year":"2013","journal-title":"Trends Microbiol."},{"key":"ref_33","first-page":"867","article-title":"Germination and inactivation of Bacillus subtilis induced by moderate hydrostatic pressure","volume":"107","author":"Dantigny","year":"2010","journal-title":"Biotechnol. Bioeng."},{"key":"ref_34","first-page":"108","article-title":"The effect of temperature on the survival of microorganisms in a deep space vacuum","volume":"3","author":"Hagen","year":"1971","journal-title":"Space Life Sci."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"218","DOI":"10.1016\/j.fm.2014.07.022","article-title":"Survival of Clostridium difficile spores at low temperatures","volume":"46","author":"Deng","year":"2015","journal-title":"Food Microbiol."}],"container-title":["Sensors"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8220\/20\/15\/4150\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T09:51:48Z","timestamp":1760176308000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8220\/20\/15\/4150"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,7,26]]},"references-count":35,"journal-issue":{"issue":"15","published-online":{"date-parts":[[2020,8]]}},"alternative-id":["s20154150"],"URL":"https:\/\/doi.org\/10.3390\/s20154150","relation":{},"ISSN":["1424-8220"],"issn-type":[{"type":"electronic","value":"1424-8220"}],"subject":[],"published":{"date-parts":[[2020,7,26]]}}}