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Biobanks have become an integral part of health and bioscience research. However, the ultra-low temperature (ULT) storage methods that biobanks employ [ULT freezers and liquid nitrogen (LN2)] are associated with carbon emissions that contribute to anthropogenic climate change. This paper aims to provide a 'Roadmap' for reducing carbon emissions associated with ULT storage in biobanking. The Roadmap offers recommendations associated with nine areas of ULT storage practice: four relating to ULT freezers, three associated with LN2 storage, and two generalised discussions regarding biosample management and centralisation. For each practice, we describe (a) the best approaches to mitigate carbon emissions, (b) explore barriers associated with hindering their implementation, and (c) make a series of recommendations that can help biobank stakeholders overcome these barriers. The recommendations were the output of a one year, UK-based, multidisciplinary research project that involved a quantitative Carbon Footprinting Assessment of the emissions associated with 1 year of ULT storage (for both freezers and LN2) at four different case study sites; as well as two follow up stakeholder workshops to qualitatively explore UK biobank stakeholder perceptions, views, and experiences on how to consider such assessments within the broader social, political, financial, technical, and cultural contexts of biobanking. [ABSTRACT FROM AUTHOR]

Abstract Biobanks have become an integral part of health and bioscience research. However, the ultra-low temperature (ULT) storage methods that biobanks employ [ULT freezers and liquid nitrogen (LN2)] are associated with carbon emissions that contribute to anthropogenic climate change. This paper aims to provide a ‘Roadmap’ for reducing carbon emissions associated with ULT storage in biobanking. The Roadmap offers recommendations associated with nine areas of ULT storage practice: four relating to ULT freezers, three associated with LN2 storage, and two generalised discussions regarding biosample management and centralisation. For each practice, we describe (a) the best approaches to mitigate carbon emissions, (b) explore barriers associated with hindering their implementation, and (c) make a series of recommendations that can help biobank stakeholders overcome these barriers. The recommendations were the output of a one year, UK-based, multidisciplinary research project that involved a quantitative Carbon Footprinting Assessment of the emissions associated with 1 year of ULT storage (for both freezers and LN2) at four different case study sites; as well as two follow up stakeholder workshops to qualitatively explore UK biobank stakeholder perceptions, views, and experiences on how to consider such assessments within the broader social, political, financial, technical, and cultural contexts of biobanking.

In this study, short-term liquid nitrogen (LN) storage was used as a strategy to conserve Pyrostegia venusta embryos, and its effects on in vitro germination, phenotypic and biochemical characteristics, and in vitro secondary metabolite production were assessed. Embryos stored in LN for 1 and 7 days presented a higher germination rate and germination speed index compared to those of the control (non-cryostored embryos). Short-term LN storage also favored the phenotypic characteristics of seedlings. LN storage significantly affected the proteins (PTN), soluble sugar (SS) and reducing sugar (RS) contents, oxidative metabolism, and phenylalanine ammonia-lyase (PAL) activity, as well as the total phenolic compound, flavonoid, phytosterol, and alkaloid levels in seedlings regenerated from embryos cryostored for 7 days. Benzoic acid derivatives and flavonoids were observed in regenerated non-acclimatized seedlings. LN storage did not affect the survival rate or phenotypic characteristics of seedlings during acclimatization. Acclimatization promoted significant changes in PTN, SS and RS contents, oxidative metabolism, and PAL activity in seedlings from embryos cryostored for 7 days. Roots from acclimatized seedlings exhibited the highest phenolic, phytosterol, and total alkaloid levels. Differences in the chromatographic profiles of the acclimatized seedlings compared with the non-acclimatized seedlings were observed. LN storage can be an effective means of ex situ conservation of P. venusta genetic resources. [ABSTRACT FROM AUTHOR]

In this study, short-term liquid nitrogen (LN) storage was used as a strategy to conserve Pyrostegia venusta embryos, and its effects on in vitro germination, phenotypic and biochemical characteristics, and in vitro secondary metabolite production were assessed. Embryos stored in LN for 1 and 7 days presented a higher germination rate and germination speed index compared to those of the control (non-cryostored embryos). Short-term LN storage also favored the phenotypic characteristics of seedlings. LN storage significantly affected the proteins (PTN), soluble sugar (SS) and reducing sugar (RS) contents, oxidative metabolism, and phenylalanine ammonia-lyase (PAL) activity, as well as the total phenolic compound, flavonoid, phytosterol, and alkaloid levels in seedlings regenerated from embryos cryostored for 7 days. Benzoic acid derivatives and flavonoids were observed in regenerated non-acclimatized seedlings. LN storage did not affect the survival rate or phenotypic characteristics of seedlings during acclimatization. Acclimatization promoted significant changes in PTN, SS and RS contents, oxidative metabolism, and PAL activity in seedlings from embryos cryostored for 7 days. Roots from acclimatized seedlings exhibited the highest phenolic, phytosterol, and total alkaloid levels. Differences in the chromatographic profiles of the acclimatized seedlings compared with the non-acclimatized seedlings were observed. LN storage can be an effective means of ex situ conservation of P. venusta genetic resources.

Cryopreservation of clonal plant germplasm is a reliable way to preserve important agronomic traits and protect against loss of crop genetic diversity of many horticultural species. Dormant bud cryopreservation techniques present an efficient alternative to the labor-intensive shoot tip cryopreservation process and may allow a single technician to preserve large quantities of germplasm in a season. This method of cryopreservation takes advantage of the natural dormancy in cold hardy crops, making it a viable technique for only deciduous trees and shrubs. Many factors must be considered when attempting to perform dormant bud research methods to applied-level germplasm preservation efforts. This process is necessarily a seasonal endeavor, which puts strain on labor and facilities particularly in winter. Integration of methods and procedures using different crop species or new equipment provides additional challenges that must be tested in advance. By identifying variables of dormant bud processing in cryopreservation literature, options emerge that allow for the modification of reported methods to work within the confines of institutional resources. Infrastructure, pre-processing, and recovery stages are discussed in terms of necessity and available alternatives to allow informed decision making in establishing an applied dormant budwood genebank. Key message: This review aims to provide information about the DB cryopreservation method with a focus on its many variables and permutations reported in literature to allow genebanks and germplasm managers to make informed decisions regarding the designation of critical resources. [ABSTRACT FROM AUTHOR]

Owing to the quick genetic turnover of the pig industry, most AI-boar sires live 2-3 yr, a period during which for 1-2 yr their semen is extended and used in liquid form for AI. Despite showing low cryosurvival, affecting fertility after AI, boar semen is frozen for easiness of transport overseas and reposition of valuable genetics. For the latter, semen is stored in liquid nitrogen (LN 2 , cryostorage) for many years, a controversial practice. Here we studied how length of cryostorage could affect sperm quality. Straws (0.5 mL) frozen using the same cryopreservation protocol at one specific location from AI- sires of proven fertility were stored in LN 2 for up to 8 yr. Post-thaw sperm quality was evaluated after 2, 4 or 8 yr of cryostorage, always compared to early thawing (15 d after freezing). Sperm motility and kinematics were evaluated post-thaw using CASA and sperm viability was cytometrically evaluated using specific fluorophores. Sperm viability was not affected by length of cryostorage, but total and progressive sperm motility were lower (p < 0.01) in sperm samples cryostored for 4 or 8 yr compared to those thawed 15 d after freezing. Cryostorage time affected sperm kinetics, but with greater intensity in the samples cryostored for 4 yr (p < 0.001) than in those for 2 yr (p < 0.01). The fact that the major phenotypic characteristic of boar spermatozoa, motility, is constrained by time of cryostorage should be considered when building cryobanks of pig semen. Attention should be placed on the finding that >2 yr of cryostorage time can be particularly detrimental for the post-thaw motility of some sires, which might require increasing sperm numbers for AI. [ABSTRACT FROM AUTHOR]

The application of composites to cryotanks has been one of the major concerns for lightweight launch vehicles. In this study, a prototype of a Type 3 cryotank was fabricated with the composite developed for cryogenic application and aluminum liner, and the cryogenic conditions were applied by filling the prototype with liquid nitrogen and then pressurizing it with gaseous nitrogen. During the experiment, delamination inside the cryotank happened. This article describes several attempts made to investigate failure through both analytical approach with thermoelastic analysis accompanied by progressive failure and experimental approach with LN2 immersion of composite/aluminum ring specimens. [ABSTRACT FROM AUTHOR]

Germplasm collections of vegetatively propagated crops are usually maintained as plants in fields or potted in greenhouses or screened enclosures. Safety duplication of these collections, as duplicate plants or separate collections, is costly and requires large amounts of space. Cryopreservation techniques which were recently developed for long-term storage of pear germalasm may offer an efficient alternative to conventional germplasm collection maintenance. Pear ( Pyrus L.) germplasm may now be stored as seeds (species), dormant buds or pollen from field-grown trees, or shoot tips from in vitro-grown plants (cultivars). Pear germplasm may now be cryopreserved and stored for long periods (> 100 yr) utilizing slow-freezing or vitrification of in vitro-grown shoot-tips. Dormant bud freezing, pollen, and seed cryopreservation of other lines are being developed to complete the base collection for Pyrus. This cryopreserved collection provides base (long-term) storage for the field-grown pear germplasm collection at the National Clonal Germplasm Repository, Corvallis, Oregon. [ABSTRACT FROM AUTHOR]