Abstract
Some microorganisms have been shown to retain a chemical signature indicative of the medium used for culturing. However, the repeatability of medium-specific chemical signatures has not been demonstrated from samples of microorganisms produced in the same batch or in different batches by the same sporulation protocol. Here, the variation in Raman spectra of bacterial endospores repeatedly prepared by the same procedure is compared to the variation between Raman spectra of spores prepared using different media. Bacillus cereus T strain (BcT) samples were correctly classified according to the medium used to induce sporulation for 100 % of spores grown in a controlled manner by the same scientist using Raman spectroscopy and multivariate data analysis. The proof-of-concept results from BcT spores produced in 12 different sporulation media showed correct classification by medium for 98 % of samples (with 100 % classification accuracy for all but one sporulation medium in this data set). Spectral differences were discerned between spores that had been freshly prepared or freeze-dried and spores that had been frozen; however, the differences did not impact the classification of the sporulation medium. Latent variables reduced the classification accuracy of BcT sporulated in G medium by different scientists using different media lots and stored for different periods of time and requires further study.
Similar content being viewed by others
References
Jackman J (2012) The microbe: the basics of structure, morphology, and physiology as they relate to microbial characterization and attribution. In: Cliff JB, Kreuzer HW, Ehrhardt CJ, Wunschel DS (eds) Chemical and physical signatures for microbial forensics. Springer, New York, pp 13–24
Jarman KH, Kreuzer-Martin HW, Wunschel DS, Valentine NB, Cliff JB, Petersen CE, Colburn HA, Wahl KL (2008) Appl Environ Microbiol 74:3573–3582
Kreuzer-Martin HW, Lott MJ, Dorigan J, Ehleringer JR (2003) Proc Natl Acad Sci U S A 100:815–819
Rieck VT, Palumbo SA, Witter LD (1973) J Gen Microbiol 74:1–8
Ehrhardt CJ, Chu V, Brown T, Simmons TL, Swan BK, Bannan J, Robertson JM (2010) Appl Environ Microbiol 76:1902–1912
Wunschel DS, Colburn HA, Fox A, Fox KF, Harley WM, Wahl JH, Wahl KL (2008) J Microbiol Methods 74:57–63
Whiteaker J, Fenselau C (2004) Agric Food Chem 53:3735–3742
DeGelder J, Scheldeman P, Leus K, Heyndrickx M, Vandenabeele P, Moens L, DeVos P (2007) Anal Bioanal Chem 389:2143–2151
DeGelder J, DeGussem K, Vandenabeele P, Vancanneyt M, DeVos P, Moens L (2007) Anal Chim Acta 603:167–175
Hutsebaut D, Maquelin K, DeVos P, Vandenabeele P, Moens L, Puppels GJ (2004) Anal Chem 2004:6274–6281
Choo-Smith L-P, Maquelin K, Vreeswijk T, Bruining HA, Puppels GJ, Thi NAN, Kirschner C, Naumann D, Ami D, Villa AM, Orsini F, Doglia SM, Lamfarraj H, Lamfarraj H, Sockalingum GD, Manfait M, Allouch P, Endtz HP (2001) Appl Environ Microbiol 67:1461–1469
Stöckel S, Meisel S, Elschner M, Rösch P, Popp J (2012) Agnew Chem Int Ed 51:5339–5342
Maquelin K, Kirschner C, Choo-Smith L-P, Van den Braak N, Endtz HP, Naumann D, Puppels GJ (2002) J Microbiol Meth 51:255–271
Maquelin K, Choo-Smith L-P, Vreeswijk T, Endtz HP, Smith B, Bennett R, Bruining HA, Puppels GJ (2000) Anal Chem 72:12–19
Helm D, Labischinski H, Schallehn G, Naumann D (1991) J Gen Microbiol 137:69–79
Kümmerle M, Scherer S, Seiler H (1998) Appl Environ Microbiol 64:2207–2214
Stöckel S, Meisel S, Elschner M, Rösch P, Popp J (2012) Anal Chem 84:9873–9880
Kelly JG, Trevisan J, Scott AD, Carmichael PL, Pollock HM, Martin-Hirsch PL, Martin FL (2011) J Proteome Res 10:1437–1448
Trevisan J, Angelov PP, Carmichael PL, Scott AD, Martin FL (2012) Analyst 137:3203–3215
Ramette A (2007) FEMS Microbiol Ecol 62:142–160
Church BD, Halvorson H, Halvorson HO (1954) J Bacteriol 68:393–399
Hashimoto T, Black SH, Gerhardt P (1960) Can J Microbiol 6:203–212
Burke WF (1982) J Gen Microbiol 128:1591–1597
Mueller JH, Hinton J (1941) Proc Soc Exp Biol and Me 48:330–333
Pisal S, Wawde G, Salvankar S, Lade S, Kadam S (2006) AAPS PharmSciTech 7:E1–E8
Schaeffer P, Millet J, Aubert JP (1965) Proc Natl Acad Sci USA 54:704–711
Morisaki S, Ota C, Matsuda K, Kaku N, Fujiwara H, Oda R, Ishibashi H, Kubo T, Kawata M (2013). J Biomed Opt 18:116011–116018
So PTC, Yew EYS, Rowlands C (2013) Biophys J 105:2641–2654
Standard Guide for Raman Shift Standards for Spectrometer Calibration (2007). ASTM Standard E1840. ASTM International, West Conshohocken, PA, 2000, DOI: 10.1520/E1840-96R07, http://www.astm.org/Standards/E1840.htm
Cozzolino D, Vadell A, Ballesteros F, Galietta G, Barlocco N (2006) Anal Bioanal Chem 385:931–939
Medina-Gutiérrez C, Luis-Quintanar J, Frausto-Reyes C, Sato-Berrú R (2005) Spectrochim Acta Part A 61:87–91
Næs T, Isaksson T, Fearn T, Davies T (2002) A user-friendly guide to multivariate calibration and classification. NIR Publications, Chicester, UK
Johnson DR, O’Higgins P, Moore WJ, McAndrew TJ (1989) Forensic Sci Int 41:41–53
Snow CC, Hartman S, Giles E, Young FA (1979) J Forensic Sci 24:448–460
Kahraman M, Keseroğlu K, Çulha M (2011) Appl Spectrosc 65:500–506
de Vries YP, Hornstra LM, de Vos WM, Abee T (2004) Appl Environ Microbiol 70:2514–2519
Huang S, Chen D, Pelczar PL, Vepachedu VR, Setlow P, Li Y (2007) J Bacteriol 189:4681–4687
Carmona P (1980) Spectrochim Acta Part A 36:705–712
Nelson WH, Dasari R, Feld M, Sperry JF (2004) Appl Spectrosc 58:1408–1412
DeGelder J, DeGussem K, Vandenabeele P, DeVos P, Moens L (2007) Anal Chim Acta 585:234–240
Bassi D, Cappa F, Cocconcelli PS (2012) Water and cations flux during sporulation and germination. In: Abel-Santos E (ed) Bacterial spores: current research and applications. Caister Academic Press, Norfolk
Bach ML, Gilvarg C (1966) J Biol Chem 241:4563–4566
Barth A, Zscherp C (2002) Q Rev Biophys 35:369–430
Aronson AI (2012) The structure and composition of the outer layers of the bacterial spores. In: Abel-Santos E (ed) Bacterial spores: current research and applications. pp 57–71
Dai D, Holder D, Raskin L, Xi C (2011). BMC Microbiol 11 (59)
Madonna AJ, Basile F, Furlong E, Voorhees KJ (2001) Rapid Commun Mass Sp 15:1068–1074
Shields MJ, Hahn KR, Janzen TW, Goji N, Thomas MC, Kingombe CB, Paquet C, Kell AJ, Amoako KK (2012) J Food Prot 75:1243–1248
Whiteaker J, Karns J, Fenselau C, Purdue ML (2004) Foodborne Pathog Dis 1:185–194
Gao W, Smith DW, Li Y (2007) Water Environ Res 79:507–513
Wyatt LR, Waites WM (1975) J Gen Microbiol 89:337–344
Harz M, Rösch P, Peschke P-D, Ronnenberger O, Burkhardt H, Popp J (2005) Analyst 130:1543–1550
Acknowledgments
We are very grateful to Jack Hietpas for independent statistical analysis and confirmation of the results presented here. We would also like to thank Jason Brewer (FBI Laboratory, Chemistry Unit) for providing assistance with the Raman instrument and Lindsay Lundberg for spore preparations. This research was supported in part by an appointment to the Visiting Scientist Program at the FBI Laboratory Division, administered by the Oak Ridge Institute of Science and Education, through an interagency agreement between the US Department of Energy and the FBI.
Disclaimer
This is publication number 14-11 of the Laboratory Division of the Federal Bureau of Investigation. Names of commercial manufacturers are provided for information only, and inclusion does not imply endorsement by the FBI or the US Government. The views expressed are those of the authors and do not necessarily reflect the official policy or position of the FBI or the U.S. Government.
Author information
Authors and Affiliations
Corresponding author
Additional information
This article is dedicated to the memory of Diane Williams, who lost a valiant battle against ovarian cancer. Diane was a very effective mentor to many Visiting Scientist Fellows and always keen to share her expertise in chemometrics and spectroscopy and engage in both scientific and cultural discussions on numerous subjects. She will be missed.
Electronic supplementary material
Below is the link to the electronic supplementary material.
ESM 1
(PDF 19 kb)
Rights and permissions
About this article
Cite this article
Dettman, J.R., Goss, J.M., Ehrhardt, C.J. et al. Forensic differentiation of Bacillus cereus spores grown using different culture media using Raman spectroscopy. Anal Bioanal Chem 407, 4757–4766 (2015). https://doi.org/10.1007/s00216-015-8677-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00216-015-8677-3