scholarly journals Alcohol keeps eDNA at the party longer

2021 ◽  
Vol 4 ◽  
Author(s):  
Sarah Licul ◽  
Rachael Impey ◽  
Andrew Weeks

For a typical eDNA water study, water will be filtered on site, before prompt transfer to a laboratory for DNA extraction and required scientific analysis. In a setting where transport is quick and available, this is a straightforward process. However, many of our studies can occur in remote Australia where sample preservation presents many logistical challenges. Typically, we advise clients to store eDNA water filters after sampling below 4 °C to ensure minimal DNA degradation. For many clients however, field studies often occur in an isolated setting without adequate refrigeration facilities, and as such present challenges for this process. Rather than compromise on sample integrity, EnviroDNA conducted a pilot study into the use of alternate preservation methods on our most commonly used 0.22 mm Sterivex filters. With help from our friendly neighbourhood goldfish tank, our standard 4 °C protocol was compared to a variety of conditions including filled ethanol filters, flushed ethanol filters, lysis buffer and silica bead storage conditions at both 4 °C and room temperature. The study, conducted at various time points over 14-days, used qPCR to quantify the amount of DNA extracted from the filter. Our results revealed that storage within or using flushed ethanol, allowed the samples to be stored for longer time intervals at room temperature, with similar, or in some cases, improved DNA elutions. This protocol optimisation has allowed us to offer an alternate sample storage protocol for clients, expanding the availability and accessibility of eDNA biodiversity assessments around Australia.

Author(s):  
M. Quan ◽  
M.S. Mulders ◽  
D.G.A. Meltzer

Investigaltions to determine the effect of sample storage on the concentration of copper in liver tissue and on the activity of erythrocyte superoxide dismutase were undertaken in preparation for a study of blesbok (Damaliscus pygargus phillipsi) that were suspected to be suffering from copper deficiency. Two liver samples were collected from each of 20 culled blesbok in a manner that simulated the collection of biopsies from the live animal. These samples were stored either in 10 % formalin or frozen at -20 °C until analysed 4 1/2 months later. The effect of different methods of sample storage on superoxide dismutase activity was determined. Erythrocytes collected from 3 Jersey cows and 5 culled blesbok were washed and divided into 0.5m portions, stored at room temperature (~20 °C), in a refrigerator (4 °C), frozen at -20 °C in a freezer, and in liquid nitrogen (-200 °C). An analysis of superoxide dismutase activity was undertaken using a commercial assay kit at intervals of 2-4 days until the levels of activity had fallen significantly. The copper concentration in formalin-preserved liver samples was significantly lower than that measured in frozen liver tissue apparently as a result of leaching. The activity of superoxide dismutase in cattle blood was unchanged for 4 days at room temperature but fell appreciably after 2 days at 4 °C and -20 °C. Enzyme activity remained unchanged for 200 days in erythrocytes stored in liquid nitrogen. Superoxide dismutase activity levels in healthy blesbok were considerably lower than those measured in Jersey cows and remained unaffected for up to 6 days in samples stored at 4 °C and 20 °C. The level of activity fell significantly thereafter. Samples stored in liquid nitrogen were unchanged after 40 days.


mSphere ◽  
2020 ◽  
Vol 5 (1) ◽  
Author(s):  
Yali Liang ◽  
Tianyu Dong ◽  
Minjian Chen ◽  
Lianping He ◽  
Tingzhang Wang ◽  
...  

ABSTRACT The contribution of human gastrointestinal (GI) microbiota and metabolites to host health has recently become much clearer. However, many confounding factors can influence the accuracy of gut microbiome and metabolome studies, resulting in inconsistencies in published results. In this study, we systematically investigated the effects of fecal sampling regions and storage and retrieval conditions on gut microbiome and metabolite profiles from three healthy children. Our analysis indicated that compared to homogenized and snap-frozen samples (standard control [SC]), different sampling regions did not affect microbial community alpha diversity, while a total of 22 of 176 identified metabolites varied significantly across different sampling regions. In contrast, storage conditions significantly influenced the microbiome and metabolome. Short-term room temperature storage had a minimal effect on the microbiome and metabolome profiles. Sample storage in RNALater showed a significant level of variation in both microbiome and metabolome profiles, independent of the storage or retrieval conditions. The effect of RNALater on the metabolome was stronger than the effect on the microbiome, and individual variability between study participants outweighed the effect of RNALater on the microbiome. We conclude that homogenizing stool samples was critical for metabolomic analysis but not necessary for microbiome analysis. Short-term room temperature storage had a minimal effect on the microbiome and metabolome profiles and is recommended for short-term fecal sample storage. In addition, our study indicates that the use of RNALater as a storage medium of stool samples for microbial and metabolomic analyses is not recommended. IMPORTANCE The gastrointestinal microbiome and metabolome can provide a new angle to understand the development of health and disease. Stool samples are most frequently used for large-scale cohort studies. Standardized procedures for stool sample handling and storage can be a determining factor for performing microbiome or metabolome studies. In this study, we focused on the effects of stool sampling regions and stool sample storage conditions on variations in the gut microbiome composition and metabolome profile.


Author(s):  
Michael Worlako Klu ◽  
John Antwi Apenteng ◽  
Bright Selorm Addy ◽  
David Ntinagyei Mintah ◽  
Elikem Katsekpor

Objective: Stability studies on flucloxacillin sodium in reconstituted oral suspensions were carried out. The experiment sought to investigate the effects that the different types of water for reconstitution and different storage conditions have on the stability of flucloxacillin sodium in the reconstituted suspensions.Methods: Suspensions of flucloxacillin sodium were reconstituted with tap water, commercial bottled water (Voltic brand was used), commercial sachet water (Everpure brand was used) treated tap water and distilled water and stored under refrigeration (RF) (4-6 °C), at room temperature (RT) (31-33 °C) and in a bowl of water (BW) (26-27 °C). Assay of flucloxacillin sodium was by iodimetry at predetermined time intervals for 8 d.Results: The amount of flucloxacillin sodium in all the suspensions stored under the various storage conditions reduced with time and at different rates. The percentage breakdown, a parameter of stability, was calculated for each reconstituted suspension stored at the different conditions investigated and they were as follows: commercial bottled water (RT-22.40 %, RF-9.90 % and BW-15.90 %), distilled water (RT-29.14 %, RF-18.0 %, BW-28.80 %), tap water (RT-25.0%, RF-14.60 % and BW-25.10 %) and commercial sachet water (RT-25.0 %, RF-10.17 % and BW-22.50 %).Conclusion: At the end of the study, it was found that those suspensions reconstituted with the commercial bottled water were the most stable and had the smallest breakdown of flucloxacillin sodium whereas those reconstituted with distilled water were the least stable and had the largest breakdown of flucloxacillin sodium. Commercial sachet water reconstituted more stable suspensions than tap water. Also, the suspensions stored under refrigeration were the most stable followed by those stored in a bowl of water. The formulations kept at room temperature were the least stable and thus, had the largest breakdown of flucloxacillin sodium.


2020 ◽  
Vol 78 (2) ◽  
pp. 529-535
Author(s):  
Marlena Walter ◽  
Jens Wiltfang ◽  
Jonathan Vogelgsang

Previous studies on blood-based biomarkers for Alzheimer’s disease suggest a less invasive blood test might be a valuable screening tool for Alzheimer-specific pathology. Pre-analytical sample storage conditions seem to play an important role on amyloid-β (Aβ) stability, impacting reliability and reproducibility. This study shows that Aβ40, Aβ42, and Aβ42/40 levels significantly and early decrease during storage at room temperature in whole blood or plasma. Storing blood samples at 4°C leads to stable Aβ peptide concentrations up to 72 h. In addition, Aβ peptides can be measured in capillary blood with a stable Aβ42/40 ratio up to 72 h at 4°C.


2009 ◽  
Vol 14 (5) ◽  
pp. 538-546 ◽  
Author(s):  
Ryan MacArthur ◽  
William Leister ◽  
Henrike Veith ◽  
Paul Shinn ◽  
Noel Southall ◽  
...  

The authors describe how room temperature storage of a 1120-member compound library prepared in either DMSO or in a hydrated-DMSO/water (67/33) mixture affects the reproducibility of potency values as monitored using cytochrome P450 1A2 and 2D6 isozyme assays. The bioluminescent assays showed Z′ factors of 0.71 and 0.62, with 17% and 32% of the library found as active against the CYP 1A2 and 2D6 isozymes, respectively. The authors tested the library using quantitative high-throughput screening to generate potency values for every library member, which was measured at 7 time intervals spanning 37 weeks. They calculated the minimum significant ratio (MSR) from these potency values at each time interval and found that for the library stored in DMSO, the CYP 1A2 and 2D6 assay MSRs progressed from approximately 2.0 to 5.0. The hydrated conditions showed similar performance in both MSR progression and analytical quality control results. Based on this study, the authors recommend that DMSO samples be stored in 1536-well plates for <4 months at room temperature. Furthermore, the study illustrates the degree and time scale of apparent compound potency changes due to sample storage. ( Journal of Biomolecular Screening 2009:538-546)


1992 ◽  
Vol 27 (4) ◽  
pp. 833-844 ◽  
Author(s):  
Micheline Hanna

Abstract In order to quantitatively assess the effect of sample storage conditions on the body burden analysis of organic contaminants, a comparative analysis was carried out on the unionid mussel Elliptic complanata. The mussels were divided into two groups, each with distinct storage conditions, while Group A was kept in the freezer at −20°C, Group B was kept in the refrigerator for five days at 5°C. All the compounds present in the control were also present in Group B samples. Analysis of the organic contaminants in each of these two groups showed that for total PCB concentrations, the two treatments were not significantly different; however when compared individually 6 of the 13 PCB congeners showed significant differences. The observed differences were relatively small for individual PCB congeners (7.1 to 15.3%), higher for chlorobenzenes (10.5 to 36.4%), and yet higher for HCE (44.1%); the difference for HCE, although large is nevertheless not significant, even if only marginally so.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Liliana G. Fidalgo ◽  
Mário M. Q. Simões ◽  
Susana Casal ◽  
José A. Lopes-da-Silva ◽  
Ivonne Delgadillo ◽  
...  

AbstractHyperbaric storage at room temperature (HS/RT: 75 MPa/25 °C) of vacuum-packaged fresh Atlantic salmon (Salmo salar) loins was studied for 30 days and compared to atmospheric pressure at refrigerated temperatures (AP/5 °C, 30 days) and RT (AP/25 °C, 5 days). Most of the fatty acids were not affected by storage conditions, with only a slight decrease of docosahexaenoic acid (DHA) content (n-3 polyunsaturated fatty acid) for AP samples, reflected in the lower polyene index values obtained and higher oxidation extent. For HS, a lower lipid oxidation extension and a slower increase of myofibrillar fragmentation index values were observed, when compared to AP samples. The volatile profile was similar for the HS and fresh samples, with the HS samples retaining fresh-like alcohols and aldehydes components, which disappeared in AP samples, mainly in AP/25 °C samples. The volatile profile for AP samples (5 and 25 °C) revealed mostly spoilage-like compounds due to microbial activity. Drip loss increased progressively during the 30 days of storage under HS, while a slight decrease of water holding capacity after 5 days was observed, increasing further after 30 days. Regarding textural properties, only resilience was affected by HS, decreasing after 30 days. So, HS/RT could represent an interesting extended preservation methodology of fresh salmon loins, since allows retaining important physicochemical properties for at least 15 days, while refrigeration after 5 days showed already volatile spoilage-like compounds due to microbial activity. Furthermore, this methodology allows additional considerable energy savings when compared to refrigeration.


2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Michael J. Allison ◽  
Jessica M. Round ◽  
Lauren C. Bergman ◽  
Ali Mirabzadeh ◽  
Heather Allen ◽  
...  

Abstract Objective Silica gel beads have promise as a non-toxic, cost-effective, portable method for storing environmental DNA (eDNA) immobilized on filter membranes. Consequently, many ecological surveys are turning to silica bead filter desiccation rather than ethanol preservation. However, no systematic evaluation of silica bead storage conditions or duration past 1 week has been published. The present study evaluates the quality of filter-immobilized eDNA desiccated with silica gel under different storage conditions for over a year using targeted quantitative real-time polymerase chain reaction (qPCR)-based assays. Results While the detection of relatively abundant eDNA target was stable over 15 months from either ethanol- or silica gel-preserved filters at − 20 and 4 °C, silica gel out-performed ethanol preservation at 23 °C by preventing a progressive decrease in eDNA sample quality. Silica gel filter desiccation preserved low abundance eDNA equally well up to 1 month regardless of storage temperature (18, 4, or − 20 °C). However only storage at − 20 °C prevented a noticeable decrease in detectability at 5 and 12 months. The results indicate that brief storage of eDNA filters with silica gel beads up to 1 month can be successfully accomplished at a range of temperatures. However, longer-term storage should be at − 20 °C to maximize sample integrity.


Author(s):  
O.O Fadipe

The study investigated the characteristics of packaged water stored under ambient and sunlight conditions. This is with a view to testing the effect of prolonged storage under different storage conditions on its quality. In addition it analyzed the interactions between the parameters. Two packs each of bottled and sachet water was purchased from each factory at the point of production and ready for distribution to wholesalers. Twenty eight pieces of packaged water from each factory were kept at room temperature and the same quantity were kept under sunlight. Physico-chemical and microbiological analyses were carried out on the remaining packaged water within 24hrs. Half of the samples stored at the two storage conditions were removed for analysis at 3 weeks while the remaining half was analyzed after 6weeks. The physico-chemical characteristics were within the WHO recommended values except for the pH of some samples that have values in the acidic range of 6.2-6.48. All the physico-chemical values increased for samples kept under sunlight. All the water samples showed growth in faecal coliform (4-46 cfu/100 mL) and E. coli (0-13 cfu/100 mL) for samples kept under sunlight at three weeks and this growth increased to the sixth week. The presence of E. coli is an indication that the packaged water is not pure. Displaying packaged water under the sunlight and storing beyond 3 weeks by vendors have effect on the potability of the product. The regulatory bodies should raise awareness and ensure manufacturer have a quality control unit to test on a routine basis.


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