scholarly journals Effect of storage conditions on SARS-CoV-2 RNA quantification in wastewater solids

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e11933
Author(s):  
Adrian Simpson ◽  
Aaron Topol ◽  
Bradley J. White ◽  
Marlene K. Wolfe ◽  
Krista R. Wigginton ◽  
...  

SARS-CoV-2 RNA in wastewater settled solids is associated with COVID-19 incidence in sewersheds and therefore, there is a strong interest in using these measurements to augment traditional disease surveillance methods. A wastewater surveillance program should provide rapid turn around for sample measurements (ideally within 24 hours), but storage of samples is necessary for a variety of reasons including biobanking. Here we investigate how storage of wastewater solids at 4 °C, −20 °C, and −80 °C affects measured concentrations of SARS-CoV-2 RNA. We find that short term (7 or 8 d) storage of raw solids at 4 °C has little effect on measured concentrations of SARS-CoV-2 RNA, whereas longer term storage at 4 °C (35–122 d) or freezing reduces measurements by 60%, on average. We show that normalizing SARS-CoV-2 RNA concentrations by concentrations of pepper mild mottle virus (PMMoV) RNA, an endogenous wastewater virus, can correct for changes during storage as storage can have a similar effect on PMMoV RNA as on SARS-CoV-2 RNA. The reductions in SARS-CoV-2 RNA in solids during freeze thaws is less than those reported for the same target in liquid influent by several authors.

2021 ◽  
Author(s):  
Adrian Simpson ◽  
Aaron Topol ◽  
Bradley White ◽  
Marlene Wolfe ◽  
Krista Wigginton ◽  
...  

SARS-CoV-2 RNA in wastewater settled solids is associated with COVID-19 incidence in sewersheds and therefore, there is a strong interest in using these measurements to augment traditional disease surveillance methods. A wastewater surveillance program should provide rapid turn around for sample measurements (ideally within 24 hours), but storage of samples is necessary for a variety of reasons including biobanking. Here we investigate how storage of wastewater solids at 4C, -20C, and -80C affects measured concentrations of SARS-CoV-2 RNA. We find that short term (7-8 d) storage of raw solids at 4C has little effect on measured concentrations of SARS-CoV-2 RNA, whereas longer term storage at 4C (35-122 d) or freezing reduces measurements by 60%, on average. We show that normalizing SARS-CoV-2 RNA concentrations by concentrations of pepper mild mottle virus (PMMoV) RNA, an endogenous wastewater virus, can correct for changes during storage as storage can have a similar effect on PMMoV RNA as on SARS-CoV-2 RNA. The reductions in SARS-CoV-2 RNA in solids during freeze thaws is less than those reported for the same target in liquid influent by several authors.


Forests ◽  
2021 ◽  
Vol 12 (5) ◽  
pp. 528
Author(s):  
Jelena Kranjec Orlović ◽  
Damir Drvodelić ◽  
Marko Vukelić ◽  
Matea Rukavina ◽  
Danko Diminić ◽  
...  

When natural regeneration of Quercus robur stands is hampered by an insufficient acorn yield, human assisted sowing of acorns collected in non-affected stands and stored for some period of time is performed. To inhibit the development of fungi and acorn deterioration during storage, thermotherapy is usually applied by submerging acorns for 2.5 h in water heated to 41 °C. This research aimed to test the effect of four thermotherapy treatments of different durations and/or applied temperatures as well as short-term storage at −1 °C or 3 °C on acorn internal mycobiota and germination. Fungal presence in cotyledons was analyzed in 450 acorns by isolation of mycelia on artificial media, followed by a DNA-based identification. Germination of 2000 acorns was monitored in an open field trial. Thermotherapy significantly decreased fungal diversity, while storage at 3 °C increased the isolation frequency of several fungi, mainly Penicillium spp. The most frequently isolated fungi did not show a negative impact on acorn germination after short-term storage. The study confirmed the efficiency of thermotherapy in the eradication of a part of acorn internal mycobiota, but also its effect on the proliferation of fast-colonizing fungi during storage. However, the latter showed to be more stimulated by storage conditions, specifically by storage at 3 °C.


HortScience ◽  
2019 ◽  
Vol 54 (1) ◽  
pp. 143-148 ◽  
Author(s):  
Brianna L. Ewing ◽  
Gregory M. Peck ◽  
Sihui Ma ◽  
Andrew P. Neilson ◽  
Amanda C. Stewart

Hard cider production in the United States has increased dramatically during the past decade, but there is little information on how harvest and postharvest practices affect the chemistry of the resulting cider, including concentrations of organoleptically important flavanols. For 2 years we assessed fruit, juice, and cider from a total of five apple (Malus ×domestica Borkh.) cultivars in two experiments: sequential harvests and postharvest storage. Different cultivars were used in 2015 and 2016 with the exception of ‘Dabinett’, which was assessed in both years. There were no differences in polyphenol concentrations in cider made from fruit that was harvested on three separate occasions over a 4-week period in either 2015 or 2016. Fruit storage durations and temperatures had little influence on the chemistry when the experiment was conducted in 2015, but polyphenol concentration was greater after storage in the 2016 experiment. In 2016, total polyphenols in ‘Dabinett’ ciders were 51% greater after short-term storage at 10 °C and 67% greater after long-term storage at 1 °C than the control, which was not subjected to a storage treatment. In 2016, total polyphenols in ‘Binet Rouge’ ciders were 67% greater after short-term storage at 10 °C and 94% greater after long-term storage at 1 °C than the control. Although results varied among cultivars and harvest years, storing apples for longer periods of time and at warmer temperatures may be a strategy to increase polyphenol, particularly flavanol, concentrations in hard cider.


2019 ◽  
Vol 60 (6) ◽  
pp. 925-934 ◽  
Author(s):  
Ewa Dziedzic ◽  
Jan Błaszczyk

Abstract Fruits of the sweet cherry cultivar ‘Regina’ collected from trees growing on seven rootstocks were stored in a cold room at 2 °C with a normal (NA) and controlled atmosphere (15% and 20% CO2 and 5% O2—CA1 and CA2) for 2 weeks. The rootstocks on which the trees grew and the storage conditions significantly affected all fruit parameters tested during both years of the experiment. Fruit from Damil rootstock exhibited higher mean firmness than fruit from Colt rootstock. The effect of rootstocks on the value of soluble solids content (SSC) varied, wherein the fruits from Tabel Edabriz and Damil were characterized by high SSC mean content. The organic acids content (TA) was significantly lower after storage than during harvest time. Fruits from Tabel Edabriz trees were characterized by faster ripening, as was evident by the higher SSC to TA ratio. The amount of mass lost depended significantly and only on the storage conditions—sweet cherries from both CA combinations had the lowest mass losses. The percentage of fruits showing disease symptoms was largely dependent on the weather conditions in the orchard the year before the fruit harvest, as well as atmosphere composition and RH during fruit storage. Cold storage conditions with a high (20%) CO2 content are recommended for the short-term storage of sweet cherry fruits because they preserve fruit quality parameters: a low decrease in firmness, maintenance of a high SSC/TA ratio, a low percent of fungal infections, and good preservation of green color in the peduncle.


1996 ◽  
Vol 121 (6) ◽  
pp. 1063-1068 ◽  
Author(s):  
Richard N. Arteca ◽  
Jeannette M. Arteca ◽  
Tzann-Wei Wang ◽  
Carl D. Schlagnhaufer

The purpose of this study was to evaluate physiological, biochemical, and molecular changes that occur in unrooted Pelargonium ×hortorum cuttings during storage. Pelargonium cuttings of `Sincerity' (good shipper), `Wendy Ann' (moderate shipper) and `Snowmass' (poor shipper) were stored at 25 °C and evaluated over a 5-day period. Following removal from storage, cuttings of all cultivars exhibited steady and significant decline in photosynthesis, respiration, carbohydrate, starch, and protein over time. However, no significant differences were observed among cultivars for all of these parameters. Ethylene levels produced by `Sincerity' and `Wendy Ann' began to increase 3 days following storage; whereas, `Snowmass' showed an increase after 1 day, reaching a peak at 3 days, and then declined. When unrooted cuttings of `Snowmass' were stored for 5 days at temperatures ranging from 4 to 25 °C, it was observed that those stored at 4 °C had a significantly higher visual rating, chlorophyll content, and root and shoot weight than at higher temperatures tested. As temperature increased from 10 to 25 °C, quality of cuttings declined. Changes in gene expression of two ACC synthases and an ACC oxidase were evaluated in `Snowmass' cuttings stored at 4 and 25 °C. Correlations between ethylene and ACC levels with gene expression were observed. Chemical name used: 1-aminocyclopropane-1-carboxylic acid (ACC).


Biology ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 806
Author(s):  
Aitor Fernandez-Novo ◽  
Sergio Santos-Lopez ◽  
Clara Barrajon-Masa ◽  
Patricia Mozas ◽  
Eduardo de Mercado ◽  
...  

CASA kinetic parameters are often evaluated in a diagnostic centre. How storage conditions affect ejaculates up to evaluation is unclear. We assessed, in 25 commercial bulls electroejaculated in the field, the impact of time until evaluation (0–2 h, 4–6 h, and 24 h post-ejaculation), holding temperature (5 °C vs. room temperature), and extender (AndroMed®, BIOXcell® or INRA96®) on CASA kinetic parameters. Total and progressive motility, VCL, VAP, VCL, ALH, BCF, STR, LIN, and WOB were assessed. CASA kinetic parameters were preserved for up to 4–6 h post-ejaculation, except for AndroMed®. Regardless of extender or temperature, motility decreased from 4–6 h up to 24 h, with the best values obtained with BIOXcell® at 5 °C. Our results suggest that BIOXcell® can preserve sperm motility for up to 6 h, either at 5 °C or room temperature, and also INRA96® at room temperature, with motility assessments and the percentage of the most rapid sperms being the lowest with INRA96® at 5 °C. The kinetic parameters decreased when analyses were performed at 24 h. Therefore, we suggest evaluating seminal quality as soon as possible, before 6 h after collection. These results help to fix adequate protocols for the short-term storage and shipment of bovine semen collected under field conditions.


HortScience ◽  
1992 ◽  
Vol 27 (7) ◽  
pp. 798-800 ◽  
Author(s):  
M.P. Kaczperski ◽  
A.M. Armitage

The effects of storage conditions before transplanting were examined for Petunia × hybrida Vilm. `Supercascade Lilac', viola × wittrockiana Gams `Universal Beaconsfield', and Salvia splendens F. Sellow ex Roem. & Schult `Red Hot Sally'. Plug grown seedlings were stored for 0, 7, 14, or 21 days at 5 or 10C and with continuous irradiance levels from incandescent bulbs at 0, 2, or 12 μmol·m-2·s-1. A second group was stored at 18C with irradiance from fluorescent bulbs at 105 μmol·m-2·s-1 and a 16-hour photoperiod for the same durations. Temperature was more important than irradiance in maintaining a commercially acceptable plant during the storage period. Petunia and pansy could be stored successfully for 21 days at 5 or 10C with no appreciable loss of quality; salvia could be stored for a minimum of 14 days. Seedlings of all species elongated excessively when stored >7 days at 18C and 105 μmol·m-2·s-1 irradiance. After 14 days of storage, petunias stored at 18C flowered sooner than those stored at 5 or 10C but time in a production environment (days to flower - days in storage) was similar for petunias stored at 5 or 18C.


HortScience ◽  
1996 ◽  
Vol 31 (4) ◽  
pp. 637a-637
Author(s):  
Richard N. Arteca ◽  
Jeannette M. Arteca ◽  
Tzann-Wei Wang ◽  
Carl D. Schlagnhaufer

The purpose of this study was to evaluate physiological, biochemical and molecular changes which occur in unrooted Pelargonium ×hortorum cuttings during storage. Pelargonium cuttings of `Sincerity' (good shipper), `Wendy Ann' (moderate shipper), and `Snowmass' (poor shipper) were stored at 25°C and evaluated over a 5-day period. Following removal from storage, cuttings exhibited progressive declines in photosynthesis, respiration, carbohydrate, starch and protein over time which was significant in all three cultivars, however there was little difference among the cultivars. Ethylene levels produced by `Sincerity' and `Wendy Ann' began to increase 3 days following the initiation of storage, whereas `Snowmass' showed an increase after one day reaching a peak at 3 days and was followed by a sharp decline. When unrooted cuttings of `Snowmass' were stored for a 5-day period at temperatures from 4 to 25°C, it was observed that those stored at 4°C had a significantly higher visual rating, chlorophyll content, root and shoot weight than at higher temperatures tested. The decline in quality progressively became greater from 10 to 25°C. Changes in gene expression of two ACC synthases and an ACC oxidase were evaluated in `Snowmass' cuttings which were stored at 4 and 25°C. Correlations between ethylene and ACC levels with gene expression were observed.


2020 ◽  
Vol 50 (4) ◽  
pp. 335-338
Author(s):  
Érica MANGARAVITE ◽  
Vanessa TERRA ◽  
Eric Koiti Okiyama HATTORI ◽  
Thaís Carolina da Silva DAL’SASSO ◽  
Leonardo Lopes BHERING ◽  
...  

ABSTRACT Many tropical trees have high canopies and their leaves are not accessible. Thus, the use of tissue from a more accessible organ (cambium) for DNA extraction may be an alternative for molecular studies. We adapted a feasible methodology for extracting genomic DNA from cambium tissue harvested in the field for the assessment with PCR. We tested three storage conditions (two buffers and a silica gel) and four periods of time after harvest. We used previously described protocols and tested them on three species that occur in Amazonian forests and other biomes: Anadenanthera peregrina var. peregrina, Cedrela fissilis, and Ceiba speciosa. Our protocol obtained suitable PCR-grade genomic DNA for DNA sequencing and microsatellite genotyping. We recommend the use of silica for long-term storage and the buffer with ascorbic acid for short-term storage.


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