Factors determining microbial colonization of liquid nitrogen storage tanks used for archiving biological samples

Publikation: Beitrag in FachzeitschriftArtikelForschungPeer-Review

Autoren

  • F. Bajerski
  • A. Bürger
  • Birgit Glasmacher
  • E. R.J. Keller
  • K. Müller
  • K. Mühldorfer
  • M. Nagel
  • H. Rüdel
  • T. Müller
  • J. Schenkel
  • J. Overmann

Organisationseinheiten

Externe Organisationen

  • Leibniz-Institut DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH
  • Helmholtz Zentrum München - Deutsches Forschungszentrum für Gesundheit und Umwelt
  • Leibniz-Institut für Pflanzengenetik und Kulturpflanzenforschung (IPK)
  • Leibniz-Institut für Zoo- u Wildtierforschung (IZW)
  • Fraunhofer-Institut Molekularbiologie und Angewandte Oekologie IME
  • BioKryo GmbH
  • Deutsches Krebsforschungszentrum (DKFZ)
  • Ruprecht-Karls-Universität Heidelberg
  • Technische Universität Braunschweig
Forschungs-netzwerk anzeigen

Details

OriginalspracheEnglisch
Seiten (von - bis)131-144
Seitenumfang14
FachzeitschriftApplied Microbiology and Biotechnology
Jahrgang104
Ausgabenummer1
Frühes Online-Datum28 Nov. 2019
PublikationsstatusVeröffentlicht - Jan. 2020

Abstract

The availability of bioresources is a precondition for life science research, medical applications, and diagnostics, but requires a dedicated quality management to guarantee reliable and safe storage. Anecdotal reports of bacterial isolates and sample contamination indicate that organisms may persist in liquid nitrogen (LN) storage tanks. To evaluate the safety status of cryocollections, we systematically screened organisms in the LN phase and in ice layers covering inner surfaces of storage tanks maintained in different biobanking facilities. We applied a culture-independent approach combining cell detection by epifluorescence microscopy with the amplification of group-specific marker genes and high-throughput sequencing of bacterial ribosomal genes. In the LN phase, neither cells nor bacterial 16S rRNA gene copy numbers were detectable (detection limit, 102 cells per ml, 103 gene copies per ml). In several cases, small numbers of bacteria of up to 104 cells per ml and up to 106 gene copies per ml, as well as Mycoplasma, or fungi were detected in the ice phase formed underneath the lids or accumulated at the bottom. The bacteria most likely originated from the stored materials themselves (Elizabethingia, Janthibacterium), the technical environment (Pseudomonas, Acinetobacter, Methylobacterium), or the human microbiome (Bacteroides, Streptococcus, Staphylococcus). In single cases, bacteria, Mycoplasma, fungi, and human cells were detected in the debris at the bottom of the storage tanks. In conclusion, the limited microbial load of the ice phase and in the debris of storage tanks can be effectively avoided by minimizing ice formation and by employing hermetically sealed sample containers.

ASJC Scopus Sachgebiete

Zitieren

Factors determining microbial colonization of liquid nitrogen storage tanks used for archiving biological samples. / Bajerski, F.; Bürger, A.; Glasmacher, Birgit et al.
in: Applied Microbiology and Biotechnology, Jahrgang 104, Nr. 1, 01.2020, S. 131-144.

Publikation: Beitrag in FachzeitschriftArtikelForschungPeer-Review

Bajerski, F, Bürger, A, Glasmacher, B, Keller, ERJ, Müller, K, Mühldorfer, K, Nagel, M, Rüdel, H, Müller, T, Schenkel, J & Overmann, J 2020, 'Factors determining microbial colonization of liquid nitrogen storage tanks used for archiving biological samples', Applied Microbiology and Biotechnology, Jg. 104, Nr. 1, S. 131-144. https://doi.org/10.1007/s00253-019-10242-1, https://doi.org/10.15488/9391
Bajerski, F., Bürger, A., Glasmacher, B., Keller, E. R. J., Müller, K., Mühldorfer, K., Nagel, M., Rüdel, H., Müller, T., Schenkel, J., & Overmann, J. (2020). Factors determining microbial colonization of liquid nitrogen storage tanks used for archiving biological samples. Applied Microbiology and Biotechnology, 104(1), 131-144. https://doi.org/10.1007/s00253-019-10242-1, https://doi.org/10.15488/9391
Bajerski F, Bürger A, Glasmacher B, Keller ERJ, Müller K, Mühldorfer K et al. Factors determining microbial colonization of liquid nitrogen storage tanks used for archiving biological samples. Applied Microbiology and Biotechnology. 2020 Jan;104(1):131-144. Epub 2019 Nov 28. doi: 10.1007/s00253-019-10242-1, 10.15488/9391
Bajerski, F. ; Bürger, A. ; Glasmacher, Birgit et al. / Factors determining microbial colonization of liquid nitrogen storage tanks used for archiving biological samples. in: Applied Microbiology and Biotechnology. 2020 ; Jahrgang 104, Nr. 1. S. 131-144.
Download
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abstract = "The availability of bioresources is a precondition for life science research, medical applications, and diagnostics, but requires a dedicated quality management to guarantee reliable and safe storage. Anecdotal reports of bacterial isolates and sample contamination indicate that organisms may persist in liquid nitrogen (LN) storage tanks. To evaluate the safety status of cryocollections, we systematically screened organisms in the LN phase and in ice layers covering inner surfaces of storage tanks maintained in different biobanking facilities. We applied a culture-independent approach combining cell detection by epifluorescence microscopy with the amplification of group-specific marker genes and high-throughput sequencing of bacterial ribosomal genes. In the LN phase, neither cells nor bacterial 16S rRNA gene copy numbers were detectable (detection limit, 102 cells per ml, 103 gene copies per ml). In several cases, small numbers of bacteria of up to 104 cells per ml and up to 106 gene copies per ml, as well as Mycoplasma, or fungi were detected in the ice phase formed underneath the lids or accumulated at the bottom. The bacteria most likely originated from the stored materials themselves (Elizabethingia, Janthibacterium), the technical environment (Pseudomonas, Acinetobacter, Methylobacterium), or the human microbiome (Bacteroides, Streptococcus, Staphylococcus). In single cases, bacteria, Mycoplasma, fungi, and human cells were detected in the debris at the bottom of the storage tanks. In conclusion, the limited microbial load of the ice phase and in the debris of storage tanks can be effectively avoided by minimizing ice formation and by employing hermetically sealed sample containers.",
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note = "Funding information: This study was funded by the Alliance of German Cryobanks (Gemeinschaft Deutscher Kryobanken, GDK). This work was supported by {\textquoteleft}EUCOMM: Tools for Functional Annotation of the Mouse Genome{\textquoteright} (EUCOMMTOOLS) project - grant agreement no [FP7-HEALTH-F4-2010-261492] and {\textquoteleft}ExNet-0041-Phase2-3 („SyNErgy-HMGU“){\textquoteleft} through the Initiative and Network Fund of the Helmholtz Association. Acknowledgments We gratefully acknowledge the support by Angelika Senula, Anika Methner, Doris B{\"u}chner, Julia Guewa, Katerina Zelena, Franziska Klann, Petra Henke, Javier Pascual and Johannes Sikorski for critical comments and statistical advice. Furthermore, we thank Martin Weing{\"a}rtner for taking the samples at Fraunhofer IME.",
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TY - JOUR

T1 - Factors determining microbial colonization of liquid nitrogen storage tanks used for archiving biological samples

AU - Bajerski, F.

AU - Bürger, A.

AU - Glasmacher, Birgit

AU - Keller, E. R.J.

AU - Müller, K.

AU - Mühldorfer, K.

AU - Nagel, M.

AU - Rüdel, H.

AU - Müller, T.

AU - Schenkel, J.

AU - Overmann, J.

N1 - Funding information: This study was funded by the Alliance of German Cryobanks (Gemeinschaft Deutscher Kryobanken, GDK). This work was supported by ‘EUCOMM: Tools for Functional Annotation of the Mouse Genome’ (EUCOMMTOOLS) project - grant agreement no [FP7-HEALTH-F4-2010-261492] and ‘ExNet-0041-Phase2-3 („SyNErgy-HMGU“)‘ through the Initiative and Network Fund of the Helmholtz Association. Acknowledgments We gratefully acknowledge the support by Angelika Senula, Anika Methner, Doris Büchner, Julia Guewa, Katerina Zelena, Franziska Klann, Petra Henke, Javier Pascual and Johannes Sikorski for critical comments and statistical advice. Furthermore, we thank Martin Weingärtner for taking the samples at Fraunhofer IME.

PY - 2020/1

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N2 - The availability of bioresources is a precondition for life science research, medical applications, and diagnostics, but requires a dedicated quality management to guarantee reliable and safe storage. Anecdotal reports of bacterial isolates and sample contamination indicate that organisms may persist in liquid nitrogen (LN) storage tanks. To evaluate the safety status of cryocollections, we systematically screened organisms in the LN phase and in ice layers covering inner surfaces of storage tanks maintained in different biobanking facilities. We applied a culture-independent approach combining cell detection by epifluorescence microscopy with the amplification of group-specific marker genes and high-throughput sequencing of bacterial ribosomal genes. In the LN phase, neither cells nor bacterial 16S rRNA gene copy numbers were detectable (detection limit, 102 cells per ml, 103 gene copies per ml). In several cases, small numbers of bacteria of up to 104 cells per ml and up to 106 gene copies per ml, as well as Mycoplasma, or fungi were detected in the ice phase formed underneath the lids or accumulated at the bottom. The bacteria most likely originated from the stored materials themselves (Elizabethingia, Janthibacterium), the technical environment (Pseudomonas, Acinetobacter, Methylobacterium), or the human microbiome (Bacteroides, Streptococcus, Staphylococcus). In single cases, bacteria, Mycoplasma, fungi, and human cells were detected in the debris at the bottom of the storage tanks. In conclusion, the limited microbial load of the ice phase and in the debris of storage tanks can be effectively avoided by minimizing ice formation and by employing hermetically sealed sample containers.

AB - The availability of bioresources is a precondition for life science research, medical applications, and diagnostics, but requires a dedicated quality management to guarantee reliable and safe storage. Anecdotal reports of bacterial isolates and sample contamination indicate that organisms may persist in liquid nitrogen (LN) storage tanks. To evaluate the safety status of cryocollections, we systematically screened organisms in the LN phase and in ice layers covering inner surfaces of storage tanks maintained in different biobanking facilities. We applied a culture-independent approach combining cell detection by epifluorescence microscopy with the amplification of group-specific marker genes and high-throughput sequencing of bacterial ribosomal genes. In the LN phase, neither cells nor bacterial 16S rRNA gene copy numbers were detectable (detection limit, 102 cells per ml, 103 gene copies per ml). In several cases, small numbers of bacteria of up to 104 cells per ml and up to 106 gene copies per ml, as well as Mycoplasma, or fungi were detected in the ice phase formed underneath the lids or accumulated at the bottom. The bacteria most likely originated from the stored materials themselves (Elizabethingia, Janthibacterium), the technical environment (Pseudomonas, Acinetobacter, Methylobacterium), or the human microbiome (Bacteroides, Streptococcus, Staphylococcus). In single cases, bacteria, Mycoplasma, fungi, and human cells were detected in the debris at the bottom of the storage tanks. In conclusion, the limited microbial load of the ice phase and in the debris of storage tanks can be effectively avoided by minimizing ice formation and by employing hermetically sealed sample containers.

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KW - Biobanking

KW - Cryobank

KW - Cryopreservation

KW - Microbial contamination

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