Safeguarding bacterial resources promotes biotechnological innovation

Heylen, Kim; Hoefman, Sven; Vekeman, Bram; Peiren, Jindrich; Vos, Paul De

doi:10.1007/s00253-011-3797-y

Cited by 38 publications

(39 citation statements)

References 64 publications

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“…Cryopreservation was confirmed less detrimental to sensitive bacteria than lyophilization (Heylen et al ., ), which combines freezing and drying steps, as indicated by higher viability and shorter t lag of cryopreserved compared to lyophilized samples directly after processing in the absence of cryoprotectant. During freezing, bacterial cells are exposed to two main stresses.…”

Section: Discussionmentioning

confidence: 99%

Effect of cryopreservation and lyophilization on viability and growth of strict anaerobic human gut microbes

Bircher

Geirnaert

Hammes

et al. 2018

Microbial Biotechnology

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SummaryStrict anaerobic gut microbes have been suggested as ‘next‐generation probiotics’ for treating several intestinal disorders. The development of preservation techniques is of major importance for therapeutic application. This study investigated cryopreservation (−80°C) and lyophilization survival and storage stability (4°C for 3 months) of the strict anaerobic gut microbes Bacteroides thetaiotaomicron, Faecalibacterium prausnitzii, Roseburia intestinalis, Anaerostipes caccae, Eubacterium hallii and Blautia obeum. To improve preservation survival, protectants sucrose and inulin (both 5% w/v) were added for lyophilization and were also combined with glycerol (15% v/v) for cryopreservation. Bacterial fitness, evaluated by maximum growth rate and lag phase, viability and membrane integrity were determined using a standardized growth assay and by flow cytometry as markers for preservation resistance. Lyophilization was more detrimental to viability and fitness than cryopreservation, but led to better storage stability. Adding sucrose and inulin enhanced viability and the proportion of intact cells during lyophilization of all strains. Viability of protectant‐free B. thetaiotaomicron, A. caccae and F. prausnitzii was above 50% after cryopreservation and storage and increased to above 80% if protectants were present. The addition of glycerol, sucrose and inulin strongly enhanced the viability of B. obeum, E. hallii and R. intestinalis from 0.03–2% in protectant‐free cultures to 11–37%. This is the first study that quantitatively compared the effect of cryopreservation and lyophilization and the addition of selected protectants on viability and fitness of six strict anaerobic gut microbes. Our results suggest that efficiency of protectants is process‐ and species‐specific.

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Section: Discussionmentioning

confidence: 99%

Effect of cryopreservation and lyophilization on viability and growth of strict anaerobic human gut microbes

Bircher

Geirnaert

Hammes

et al. 2018

Microbial Biotechnology

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“…In non-BRC labs the preservation method of freezing at −80°C is preferred over freeze-drying or other drying techniques, because of the direct access to electrical freezers for most researchers and the straightforwardness of the procedure [16]. To avoid cellular damage during cryopreservation and subsequent thawing, a wide array of cryoprotective agents (CPA) has been applied.…”

Section: Introductionmentioning

confidence: 99%

Optimized Cryopreservation of Mixed Microbial Communities for Conserved Functionality and Diversity

et al. 2014

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The use of mixed microbial communities (microbiomes) for biotechnological applications has steadily increased over the past decades. However, these microbiomes are not readily available from public culture collections, hampering their potential for widespread use. The main reason for this lack of availability is the lack of an effective cryopreservation protocol. Due to this critical need, we evaluated the functionality as well as the community structure of three different types of microbiomes before and after cryopreservation with two cryoprotective agents (CPA). Microbiomes were selected based upon relevance towards applications: (1) a methanotrophic co-culture (MOB), with potential for mitigation of greenhouse gas emissions, environmental pollutants removal and bioplastics production; (2) an oxygen limited autotrophic nitrification/denitrification (OLAND) biofilm, with enhanced economic and ecological benefits for wastewater treatment, and (3) fecal material from a human donor, with potential applications for fecal transplants and pre/probiotics research. After three months of cryopreservation at −80°C, we found that metabolic activity, in terms of the specific activity recovery of MOB, aerobic ammonium oxidizing bacteria (AerAOB) and anaerobic AOB (AnAOB, anammox) in the OLAND mixed culture, resumes sooner when one of our selected CPA [dimethyl sulfoxide (DMSO) and DMSO plus trehalose and tryptic soy broth (DMSO+TT)] was added. However, the activity of the fecal community was not influenced by the CPA addition, although the preservation of the community structure (as determined by 16S rRNA gene sequencing) was enhanced by addition of CPA. In summary, we have evaluated a cryopreservation protocol that succeeded in preserving both community structure and functionality of value-added microbiomes. This will allow individual laboratories and culture collections to boost the use of microbiomes in biotechnological applications.

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“…Therefore, it is important that, after growing novel microbes, researchers devise appropriate preservation protocol(s) suitable for the microorganisms. Microbial resource centers also play an important role in the development of protocols related to long-term preservation; checking the viability and authenticity of preserved cultures; providing training in areas related to microbial handling, biosafety, and biosecurity; and offering reference strains to the scientifi c community for quality control and molecular biology research (Heylen et al 2012 ). Nowadays, it is becoming very important to study microbial taxonomy, preservation, genomic studies, and microbial ecology to preserve the valuable gene pool and maintain a diversity of microorganisms for future use.…”

Section: Role In Maintaining the Microbial Resourcesmentioning

confidence: 99%

Microbiomics: An Approach to Community Microbiology

Sharma

Brahma

Sharma

et al. 2015

PlantOmics: The Omics of Plant Science

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Many biologists have known that there are many groups of microorganisms that, although they cannot be cultured, do exist and infl uence the life of mammals, plants, and other small multicellular organisms. Nowadays, many new sequencing techniques have been developed that, with very high precision, can identify the presence of these microorganisms and how they affect others. The microbiota of humans and plants can be studied, and their effects on health and growth are becoming known using the transcriptome analysis of interaction. Life on earth has evolved over billions of years, and its most initial form-the microorganism-has evolved in gradually changing environmental conditions. They are present everywhere: from high temperatures to freezing conditions, in water and in air, on

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Safeguarding bacterial resources promotes biotechnological innovation

Cited by 38 publications

References 64 publications

Effect of cryopreservation and lyophilization on viability and growth of strict anaerobic human gut microbes

Effect of cryopreservation and lyophilization on viability and growth of strict anaerobic human gut microbes

Optimized Cryopreservation of Mixed Microbial Communities for Conserved Functionality and Diversity

Microbiomics: An Approach to Community Microbiology

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