Identification and Elimination of the Clinically Relevant Multi-Resistant Environmental Bacteria <i>Ralstonia insidiosa</i> in Primary Cell Culture
In times of spreading multidrug-resistant bacteria, species identification and decontamination of cell cultures can be challenging. Here, we describe a mobile cell culture contaminant with “black dot”-like microscopic appearance in newly established irreplaceable hybridoma cell lines and its identif...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2020-10-01
|
| Series: | Microorganisms |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2076-2607/8/10/1599 |
| _version_ | 1797550679286349824 |
|---|---|
| author | Dennis Nurjadi Sébastien Boutin Katja Schmidt Melinda Ahmels Daniel Hasche |
| author_facet | Dennis Nurjadi Sébastien Boutin Katja Schmidt Melinda Ahmels Daniel Hasche |
| author_sort | Dennis Nurjadi |
| collection | DOAJ |
| description | In times of spreading multidrug-resistant bacteria, species identification and decontamination of cell cultures can be challenging. Here, we describe a mobile cell culture contaminant with “black dot”-like microscopic appearance in newly established irreplaceable hybridoma cell lines and its identification. Using 16S rRNA gene sequencing, species-specific PCRs, whole genome sequencing (WGS), and MALDI-TOF mass spectrometry, the contaminant was identified as the ubiquitous environmental and clinically relevant Gram-negative bacterium <i>Ralstonia insidiosa</i> (<i>R. insidiosa</i>), a strong biofilm producer. Further characterizations by transmission electron microscopy (TEM) and biochemical API test were not conclusive. Whole genome sequencing of our <i>R. insidiosa</i> isolate revealed numerous drug-resistance determinants. Genome-wide comparison to other <i>Ralstonia species</i> could not unambiguously designate our isolate to <i>R. insidiosa</i> (<95% average nucleotide identity) suggesting a potential novel species or subspecies, closely related to <i>R. insidiosa</i> and <i>R. pickettii</i>. After determining the antibiotic susceptibility profile, the hybridoma cell culture was successfully decontaminated with ciprofloxacin without affecting antibody production. |
| first_indexed | 2024-03-10T15:32:47Z |
| format | Article |
| id | doaj.art-b77229dbc1eb46bc8645d32394301ad7 |
| institution | Directory Open Access Journal |
| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-03-10T15:32:47Z |
| publishDate | 2020-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microorganisms |
| spelling | doaj.art-b77229dbc1eb46bc8645d32394301ad72023-11-20T17:29:12ZengMDPI AGMicroorganisms2076-26072020-10-01810159910.3390/microorganisms8101599Identification and Elimination of the Clinically Relevant Multi-Resistant Environmental Bacteria <i>Ralstonia insidiosa</i> in Primary Cell CultureDennis Nurjadi0Sébastien Boutin1Katja Schmidt2Melinda Ahmels3Daniel Hasche4Department of Infectious Diseases, Medical Microbiology and Hygiene, Heidelberg University Hospital, Im Neuenheimer Feld 324, 69120 Heidelberg, GermanyDepartment of Infectious Diseases, Medical Microbiology and Hygiene, Heidelberg University Hospital, Im Neuenheimer Feld 324, 69120 Heidelberg, GermanyGerman Cancer Research Center (DKFZ), Division of Microbiological Diagnostics (W440), Im Neuenheimer Feld 280, 69120 Heidelberg, GermanyGerman Cancer Research Center (DKFZ), Division of Viral Transformation Mechanisms (F030), Im Neuenheimer Feld 242, 69120 Heidelberg, GermanyGerman Cancer Research Center (DKFZ), Division of Viral Transformation Mechanisms (F030), Im Neuenheimer Feld 242, 69120 Heidelberg, GermanyIn times of spreading multidrug-resistant bacteria, species identification and decontamination of cell cultures can be challenging. Here, we describe a mobile cell culture contaminant with “black dot”-like microscopic appearance in newly established irreplaceable hybridoma cell lines and its identification. Using 16S rRNA gene sequencing, species-specific PCRs, whole genome sequencing (WGS), and MALDI-TOF mass spectrometry, the contaminant was identified as the ubiquitous environmental and clinically relevant Gram-negative bacterium <i>Ralstonia insidiosa</i> (<i>R. insidiosa</i>), a strong biofilm producer. Further characterizations by transmission electron microscopy (TEM) and biochemical API test were not conclusive. Whole genome sequencing of our <i>R. insidiosa</i> isolate revealed numerous drug-resistance determinants. Genome-wide comparison to other <i>Ralstonia species</i> could not unambiguously designate our isolate to <i>R. insidiosa</i> (<95% average nucleotide identity) suggesting a potential novel species or subspecies, closely related to <i>R. insidiosa</i> and <i>R. pickettii</i>. After determining the antibiotic susceptibility profile, the hybridoma cell culture was successfully decontaminated with ciprofloxacin without affecting antibody production.https://www.mdpi.com/2076-2607/8/10/1599bacterial contaminationprimary cell culture<i>Ralstonia insidiosa</i>multidrug resistance |
| spellingShingle | Dennis Nurjadi Sébastien Boutin Katja Schmidt Melinda Ahmels Daniel Hasche Identification and Elimination of the Clinically Relevant Multi-Resistant Environmental Bacteria <i>Ralstonia insidiosa</i> in Primary Cell Culture Microorganisms bacterial contamination primary cell culture <i>Ralstonia insidiosa</i> multidrug resistance |
| title | Identification and Elimination of the Clinically Relevant Multi-Resistant Environmental Bacteria <i>Ralstonia insidiosa</i> in Primary Cell Culture |
| title_full | Identification and Elimination of the Clinically Relevant Multi-Resistant Environmental Bacteria <i>Ralstonia insidiosa</i> in Primary Cell Culture |
| title_fullStr | Identification and Elimination of the Clinically Relevant Multi-Resistant Environmental Bacteria <i>Ralstonia insidiosa</i> in Primary Cell Culture |
| title_full_unstemmed | Identification and Elimination of the Clinically Relevant Multi-Resistant Environmental Bacteria <i>Ralstonia insidiosa</i> in Primary Cell Culture |
| title_short | Identification and Elimination of the Clinically Relevant Multi-Resistant Environmental Bacteria <i>Ralstonia insidiosa</i> in Primary Cell Culture |
| title_sort | identification and elimination of the clinically relevant multi resistant environmental bacteria i ralstonia insidiosa i in primary cell culture |
| topic | bacterial contamination primary cell culture <i>Ralstonia insidiosa</i> multidrug resistance |
| url | https://www.mdpi.com/2076-2607/8/10/1599 |
| work_keys_str_mv | AT dennisnurjadi identificationandeliminationoftheclinicallyrelevantmultiresistantenvironmentalbacteriairalstoniainsidiosaiinprimarycellculture AT sebastienboutin identificationandeliminationoftheclinicallyrelevantmultiresistantenvironmentalbacteriairalstoniainsidiosaiinprimarycellculture AT katjaschmidt identificationandeliminationoftheclinicallyrelevantmultiresistantenvironmentalbacteriairalstoniainsidiosaiinprimarycellculture AT melindaahmels identificationandeliminationoftheclinicallyrelevantmultiresistantenvironmentalbacteriairalstoniainsidiosaiinprimarycellculture AT danielhasche identificationandeliminationoftheclinicallyrelevantmultiresistantenvironmentalbacteriairalstoniainsidiosaiinprimarycellculture |