scholarly journals The effect of ethanol concentration on the morphological and molecular preservation of insects for biodiversity studies

2020 ◽  
Author(s):  
Daniel Marquina ◽  
Mateusz Buczek ◽  
Fredrik Ronquist ◽  
Piotr Łukasik
Keyword(s):  
Ethanol Concentration ◽  
Extended Period ◽  
Morphological Examination ◽  
Long Term Storage ◽  
Copy Numbers ◽  
Insect Biodiversity ◽  
Trade Offs ◽  
Negative Effect

Abstract1. Traditionally, insects collected for scientific purposes have been dried and pinned, or preserved in 70 % ethanol. Both methods preserve taxonomically informative exoskeletal structures well but are suboptimal for preserving DNA. Highly concentrated ethanol (95 – 100 %), preferred as a DNA preservative, has generally been assumed to make specimens brittle and prone to breaking. However, systematic studies on the correlation between ethanol concentration and specimen preservation are lacking.2. We tested how preservative ethanol concentration in combination with different sample handling regimes affect the integrity of seven insect species representing four orders, and differing substantially in the level of sclerotization. After preservation and treatments (various levels of disturbance), we counted the number of appendages (legs, wings, antennae, heads) that specimens had lost. Additionally, we assessed the preservation of DNA after long-term storage by comparing the ratio of PCR amplicon copy numbers to an added artificial standard.3. We found that high ethanol concentrations indeed induce brittleness in insects. However, the magnitude and nature of the effect varied strikingly among species. In general, ethanol concentrations at or above 90 % made the insects more brittle, but for species with robust, thicker exoskeletons, this did not translate to an increased loss of appendages. Neither freezing nor drying the insects after immersion in ethanol had a negative effect on the retention of appendages. We also found that DNA preserves less well at lower ethanol concentrations when stored at room temperature for an extended period. However, the magnitude of the effect varies among species; the concentrations at which the number of COI amplicon copies relative to the standard was significantly decreased compared to 95 % ethanol ranged from 90 % to as low as 50 %.4. While higher ethanol concentrations positively affect long-term DNA preservation, there is a clear trade-off between preserving insects for morphological examination and genetic analysis. The optimal ethanol concentration for the latter is detrimental for the former, and vice versa. These trade-offs need to be considered in large insect biodiversity surveys and other projects aiming to combine molecular work with traditional morphology-based characterization of collected specimens.

scholarly journals The effect of ethanol concentration on the morphological and molecular preservation of insects for biodiversity studies

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e10799
Author(s):  
Daniel Marquina ◽  
Mateusz Buczek ◽  
Fredrik Ronquist ◽  
Piotr Łukasik
Keyword(s):  
Ethanol Concentration ◽  
Extended Period ◽  
Morphological Examination ◽  
Long Term Storage ◽  
Copy Numbers ◽  
Insect Biodiversity ◽  
Trade Offs ◽  
Negative Effect

Traditionally, insects collected for scientific purposes have been dried and pinned, or preserved in 70% ethanol. Both methods preserve taxonomically informative exoskeletal structures well but are suboptimal for preserving DNA for molecular biology. Highly concentrated ethanol (95–100%), preferred as a DNA preservative, has generally been assumed to make specimens brittle and prone to breaking. However, systematic studies on the correlation between ethanol concentration and specimen preservation are lacking. Here, we tested how preservative ethanol concentration in combination with different sample handling regimes affect the integrity of seven insect species representing four orders, and differing substantially in the level of sclerotization. After preservation and treatments (various levels of disturbance), we counted the number of appendages (legs, wings, antennae, or heads) that each specimen had lost. Additionally, we assessed the preservation of DNA after long-term storage by comparing the ratio of PCR amplicon copy numbers to an added artificial standard. We found that high ethanol concentrations indeed induce brittleness in insects. However, the magnitude and nature of the effect varied strikingly among species. In general, ethanol concentrations at or above 90% made the insects more brittle, but for species with robust, thicker exoskeletons, this did not translate to an increased loss of appendages. Neither freezing the samples nor drying the insects after immersion in ethanol had a negative effect on the retention of appendages. However, the morphology of the insects was severely damaged if they were allowed to dry. We also found that DNA preserves less well at lower ethanol concentrations when stored at room temperature for an extended period. However, the magnitude of the effect varies among species; the concentrations at which the number of COI amplicon copies relative to the standard was significantly decreased compared to 95% ethanol ranged from 90% to as low as 50%. While higher ethanol concentrations positively affect long-term DNA preservation, there is a clear trade-off between preserving insects for morphological examination and genetic analysis. The optimal ethanol concentration for the latter is detrimental for the former, and vice versa. These trade-offs need to be considered in large insect biodiversity surveys and other projects aiming to combine molecular work with traditional morphology-based characterization of collected specimens.

scholarly journals Long-Term Cryopreservation Does Not Affect Quality of Peripheral Blood Stem Cell Grafts: A Comparative Study of Native, Short-Term and Long-Term Cryopreserved Haematopoietic Stem Cells

2021 ◽  
Vol 30 ◽  
pp. 096368972110360
Author(s):  
Daniel Lysak ◽  
Michaela Brychtová ◽  
Martin Leba ◽  
Miroslava Čedíková ◽  
Daniel Georgiev ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Statistical Significance ◽  
Extended Period ◽  
High Dose ◽  
Atp Production ◽  
Cd34 Cells ◽  
Negative Effect

Cryopreserved haematopoietic progenitor cells are used to restore autologous haematopoiesis after high dose chemotherapy. Although the cells are routinely stored for a long period, concerns remain about the maximum storage time and the possible negative effect of storage on their potency. We evaluated the effect of cryopreservation on the quality of peripheral stem cell grafts stored for a short (3 months) and a long (10 years) period and we compared it to native products.The viability of CD34+ cells remained unaffected during storage, the apoptotic cells were represented up to 10% and did not differ between groups. The clonogenic activity measured by ATP production has decreased with the length of storage (ATP/cell 1.28 nM in native vs. 0.63 in long term stored products, P < 0.05). Only borderline changes without statistical significance were detected when examining mitochondrial and aldehyde dehydrogenase metabolic activity and intracellular pH, showing their good preservation during cell storage. Our experience demonstrates that cryostorage has no major negative effect on stem cell quality and potency, and therefore autologous stem cells can be stored safely for an extended period of at least 10 years. On the other hand, long term storage for 10 years and longer may lead to mild reduction of clonogenic capacity. When a sufficient dose of stem cells is infused, these changes will not have a clinical impact. However, in products stored beyond 10 years, especially when a low number of CD34+ cells is available, the quality of stem cell graft should be verified before infusion using the appropriate potency assays.

scholarly journals How Do Trade-Offs and Synergies between Ecosystem Services Change in the Long Period? The Case Study of Uxin, Inner Mongolia, China

2019 ◽  
Vol 11 (21) ◽  
pp. 6041 ◽  
Author(s):  
Zhang ◽  
Li ◽  
Buyantuev ◽  
Bao ◽  
Zhang
Keyword(s):  
Ecosystem Services ◽  
Inner Mongolia ◽  
Stepwise Regression ◽  
Driving Forces ◽  
Extended Period ◽  
Trade Offs ◽  
Development And Utilization ◽  
Long Term Trends

Ecosystem services management should often expect to deal with non-linearities due to trade-offs and synergies between ecosystem services (ES). Therefore, it is important to analyze long-term trends in ES development and utilization to understand their responses to climate change and intensification of human activities. In this paper, the region of Uxin in Inner Mongolia, China, was chosen as a case study area to describe the spatial distribution and trends of 5 ES indicators. Changes in relationships between ES and driving forces of dynamics of ES relationships were analyzed for the period 1979–2016 using a stepwise regression. We found that: the magnitude and directions in ES relationships changed during this extended period; those changes are influenced by climate factors, land use change, technological progress, and population growth.

scholarly journals Characterization of Postharvest Fungicide-Resistant Botrytis cinerea Isolates From Commercially Stored Apple Fruit

2017 ◽  
Vol 107 (3) ◽  
pp. 362-368 ◽  
Author(s):  
Wayne M. Jurick ◽  
Otilia Macarisin ◽  
Verneta L. Gaskins ◽  
Eunhee Park ◽  
Jiujiang Yu ◽  
...  
Keyword(s):  
Botrytis Cinerea ◽  
Gray Mold ◽  
Apple Fruit ◽  
Sublethal Dose ◽  
Long Term Storage ◽  
Pose Control ◽  
First Time

Botrytis cinerea causes gray mold and is an economically important postharvest pathogen of fruit, vegetables, and ornamentals. Fludioxonil-sensitive B. cinerea isolates were collected in 2011 and 2013 from commercial storage in Pennsylvania. Eight isolates had values for effective concentrations for inhibiting 50% of mycelial growth of 0.0004 to 0.0038 μg/ml for fludioxonil and were dual resistant to pyrimethanil and thiabendazole. Resistance was generated in vitro, following exposure to a sublethal dose of fludioxonil, in seven of eight dual-resistant B. cinerea isolates. Three vigorously growing B. cinerea isolates with multiresistance to postharvest fungicides were further characterized and found to be osmosensitive and retained resistance in the absence of selection pressure. A representative multiresistant B. cinerea strain caused decay on apple fruit treated with postharvest fungicides, which confirmed the in vitro results. The R632I mutation in the Mrr1 gene, associated with fludioxonil resistance in B. cinerea, was not detected in multipostharvest fungicide-resistant B. cinerea isolates, suggesting that the fungus may be using additional mechanisms to mediate resistance. Results from this study show for the first time that B. cinerea with dual resistance to pyrimethanil and thiabendazole can also rapidly develop resistance to fludioxonil, which may pose control challenges in the packinghouse environment and during long-term storage.

Assessment of Package Performance During Long-Term Storage of Intermediate-Level Radioactive Waste

2003 ◽  
Author(s):  
A. V. Chambers ◽  
W. R. Rodwell ◽  
M. Kelly ◽  
A. R. Hoch ◽  
A. J. Baker
Keyword(s):  
Extended Period ◽  
Intermediate Level ◽  
Passive Safety ◽  
Long Term Storage ◽  
Waste Storage ◽  
Disposal Option ◽  
Intermediate Level Radioactive Waste ◽  
The Uk ◽  
The Impact

In the UK, the need for the long-term management of intermediate-level radioactive wastes arises from the absence of an established deep disposal option. As a consequence there is a requirement to demonstrate the ‘passive safety’ of waste packages in stores over a necessarily lengthy period (perhaps 150 years or more). ‘Passive safety’ is taken here to imply that the waste packages themselves would remain safe without intervention; it is assumed however, that the store building and associated infrastructure would need to be actively maintained and that the store may need to be ventilated. Importantly, methods for waste storage also need to ensure that possible future options for disposal are not foreclosed. An extended period of storage suggests that emphasis will need to be placed by waste producers on understanding a number of issues such as: • the impact of corrosion on container integrity; • the assessment of container vents and other features as a pathway for the release of radioactivity; • the extent to which wastes are conditioned for storage and the selection of appropriate methods; • the generation and behaviour of toxic or flammable gases in the storage facility; and the mitigating steps that might be needed to address adverse impacts (e.g. the type of ventilation that the store requires). In this paper, we review the requirements and current state of knowledge relevant to the assessment of operational releases of radioactive, flammable and toxic gases from wastes in long-term interim storage.

Effect of MAP on Quality of Rice for Super-Long-Term

2010 ◽  
Vol 148-149 ◽  
pp. 439-442
Author(s):  
Xi Hong Li ◽  
Jing Wang ◽  
Ling Yan Ge ◽  
Sen Wan
Keyword(s):  
Fatty Acid Content ◽  
Extended Period ◽  
Total Acid ◽  
Volatile Aldehyde ◽  
Long Term Storage ◽  
Pvc Film ◽  
Ma Packaging ◽  
Rice Samples

In this study, effects of Modified Atmosphere (MA) packaging on the quality of rice after a long term storage at room temperature have been investigated. PVC films with different thicknesses and woven bag were used as the package materials for rice samples. Results showed that MA packaging had a significant effect on the quality of the rice. After an extended period of storage, the rice in woven bag showed most severe quality deterioration compared to those stored in PVC packages. Rice packed in 0.05 mm PVC film package has the lowest value of total acid among all samples. The increase of fatty acid content in rice packed in 0.03 mm film, 0.05 mm film, 0.08 mm film and 0.12 mm film was 38.58 mg KOH/100g, 18.96 mg KOH/100g, 26.83 mg KOH/100g and 32.64 mg KOH/100g, respectively. The volatile aldehyde components accumulated in the stored rice are mainly acetaldehyde, valeraldehyde and hexanal. Small package with a MA has obvious benefits in regulating the metabolism and accumulation of volatile aldehyde components in the rice. Rice samples stored in a package of 0.05mm film had the lowest value of volatile aldehyde, while those in 0.03mm film had the highest value. This study suggests that MA packaging with a PVC film of 0.05 mm shows the best effects in maintaining the quality of rice after an extended period of storage (3 to 5 years).

scholarly journals The effects of freezing on long-term storage of canine erythrocytes

2016 ◽  
Vol 19 (2) ◽  
pp. 401-406 ◽  
Author(s):  
D. Aktaran Bala ◽  
M. Özcan
Keyword(s):  
Frozen Storage ◽  
Biochemical Properties ◽  
Quality Parameters ◽  
Consecutive Series ◽  
Time Points ◽  
Long Term Storage ◽  
Negative Effect ◽  
Natural Outcome ◽  
2,3 Dpg

Abstract Human medicine studies have so far demonstrated that erythrocytes may be preserved and stored at low temperatures for decades retaining their metabolic and biochemical properties. However, detailed studies regarding this problem are not yet available in veterinary medicine. Therefore, the objective of the current study was to investigate time-dependent effects of long-term frozen storage of canine red blood cells. Twelve healthy adult dogs meeting the criteria for blood transfusion were used in the study. Whole blood samples (450 ± 45 ml) collected from each dog were centrifuged by a cryogenic microcentrifuge and packed RBC suspensions were obtained. The samples were prewashed three times in 0.9% NaCl solution and were allocated into three groups to be evaluated at three different time points (day 0 and month 4 and 6). The samples to be frozen were subjected to glycerolization and then stored at −80°C for 4 and 6-month periods. At the end of this period the packed RBC samples were thawed, centrifuged and then washed in a consecutive series of dextrose solutions. 2,3-Diphosphoglycerate (2,3-DPG), Adenosin triphosphate (ATP), supernatant hemoglobin (SupHb), sodium (Na+) and potassium (K+) levels, residual glycerol concentrations and hemograms were evaluated and compared. Sterility tests were performed on all samples for bacterial contamination. A statistically significant decrease was noted in potassium levels, which was the natural outcome of deglycerolization process. No significant change was observed in terms of other parameters due based on different time points. In conclusion, long-term frozen storage had no negative effect on the quality parameters of canine erythrocytes.

scholarly journals Resveratrol prevents amyloid fibrillation of insulin by arresting it in a bioactive oligomeric form

2019 ◽  
Author(s):  
Bani Kumar Pathak ◽  
Debajyoti Das ◽  
Sayan Bhakta ◽  
Partha Chakrabarti ◽  
Jayati Sengupta
Keyword(s):  
Phenolic Compounds ◽  
Limiting Factor ◽  
Commercial Preparation ◽  
Long Term Storage ◽  
Natural Polyphenol ◽  
Oligomeric Form ◽  
Amyloid Fibrillation

AbstractInsulin fibrillation is a limiting factor for its long-term storage because of considerably reduced bioavailable moieties. Deposition of fibrillated insulin can also cause subcutaneous insulin amyloidoma. Toxic phenolic compounds along with Zinc are used during commercial preparation of insulin to stabilize it in a hexameric form. Designed or repurposed natural small molecules with anti-amyloidogenic properties could thus be attractive agents for preventing insulin fibrillation. Natural polyphenolic compounds which have been shown to serve as anti-amyloid agents for proteins associated with neurodegenerative diseases are potential candidates for such function.In this study we have demonstrated that resveratrol, a natural polyphenol, can not only prevent insulin fibrillation but can also preserve insulin in a bioactive oligomeric form even at high temperature. While investigating the influence of some natural polyphenols on human insulin (hINS) in a condition inductive to amyloid fibrillation at physiological pH, we found attenuation, to different extents, of insulin fibril formation. However, visualization of polyphenol-treated hINS revealed that resveratrol in particular has the unique ability to arrest hINS before the onset of fibrillation growth in soluble oligomeric forms with discrete spherical morphology. Importantly, insulin treated with resveratrol retains its full biological activity in vivo and exerts no toxicity towards cell lines. Structural characterization of the major population of resveratrol-induced insulin oligomers by cryo-EM and single particle 3D reconstruction revealed its morphology that resembled crystal structure of insulin hexamer formulated with phenolic compounds. Thus, our study suggests that resveratrol can be an effective nontoxic substituent of phenolic compounds for insulin preservation.

Characterization of aluminum nanopowders after long-term storage

2014 ◽  
Vol 321 ◽  
pp. 475-480 ◽  
Author(s):  
O.B. Nazarenko ◽  
Yu.A. Amelkovich ◽  
A.I. Sechin
Keyword(s):  
Long Term Storage ◽  
Term Storage

The Conceptual Solutions Concerning Decomissioning and Dismantling of Russian Civil Nuclear Powered Ships

2013 ◽  
Author(s):  
Konstantin N. Kulikov ◽  
Rinat A. Nizamutdinov ◽  
Andrey N. Abramov
Keyword(s):  
Economic Assessment ◽  
North West ◽  
Long Term Storage ◽  
Murmansk Region ◽  
Negative Effect ◽  
Research Procedure ◽  
State Corporation ◽  
Radiation Contamination ◽  
Selection Of

From 1959 up to 1991 nine civil nuclear powered ships were built in Russia: eight ice-breakers and one lash lighter carrier (cargo ship). At the present time three of them were taking out of service: ice-breaker “Lenin” is decommissioned as a museum and is set for storage in the port of Murmansk, nuclear ice-breakers “Arktika” and “Sibir” are berthing. The ice-breakers carrying radwastes appear to be a possible source of radiation contamination of Murmansk region and Kola Bay because the ship long-term storage afloat has the negative effect on hull’s structures. As the result of this under the auspices of the Federal Targeted Program “Nuclear and Radiation Safety of Russia for 2008 and the period until 2015” the conception and projects of decommissioning of nuclear-powered ships are developed by the State corporation Rosatom with the involvement of companies of United Shipbuilding Corporation. In developing the principal provisions of conception of decommissioning and dismantling of ice-breakers the technical and economic assessment of dismantling options in shiprepairing enterprises of North-West of Russia was performed. The paper contains description of options, research procedure, analysis of options of decommissioning and dismantling of nuclear ice-breakers, taking into account the principle of optimization of potential radioactive effect to personnel, human population and environment. The report’s conclusions contain the recommendations for selection of option for development of nuclear ice-breaker decommissioning and dismantling projects.

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