scholarly journals Evaluation of Saline as Potential Alternative to Viral Transport Media for COVID-19 Samples Stored at Different Temperatures

Author(s):  
Parul Sinha ◽  
Dinesh Kumar Jain ◽  
Sandeep Gupta ◽  
Monika Gupta ◽  
Megha Gupta ◽  
...  

Introduction: Severe acute respiratory syndrome coronavirus-2 (SARS CoV 2) virus, a causative agent of COVID-19 has led to universal pandemic. During this pandemic there has been an acute shortage of good quality Viral Transport Medium (VTM) because of increase in number of infected people worldwide. It is also difficult to maintain the transport and storing conditions in line with the guidelines in pandemics. Aim: To assess the feasibility of Oropharyngeal Swab (OP)/Nasal swabs in 0.9% normal saline in place of VTM and to analyse the effect of temperature on nucleic acid detection by rRT PCR on saline samples stored at 4ºC, ambient and at higher temperature (37ºC). Materials and Methods: The present study was an observational analytical study which included 94 positive and 5 negative samples. Patients' nasal or OP samples were collected as dry swabs and in VTM. Normal saline was added once the samples were received in the laboratory. PCR was done with saline and VTM samples both on day 1. Samples were aliquotted in 3 sets and one set was kept at 4º-8º C and other two at 25ºC and 37ºC, respectively. All positive samples were further tested on day 3, day 4 and day 6. Results were analysed and compared. Results: Samples in normal saline showed very good sensitivity at all temperatures (4º-8ºC, 25ºC and 37ºC) till day 6. Both the swab samples (in saline and in VTM) showed nearly 100% agreement in rRT-PCR results. Ct value variation was also ≤±2. Conclusion: Looking into the cost and logistics issues especially during pandemics, saline is a good and cheaper alternative to VTM and with its use, testing capacity can be expanded.

2020 ◽  
Author(s):  
Julie McAuley ◽  
Claire Fraser ◽  
Elena Paraskeva ◽  
Elizabeth Trajcevska ◽  
Michelle Sait ◽  
...  

Abstract IntroductionThe sudden arrival of the COVID-19 pandemic placed significant stresses on supply chains including viral transport medium (VTM). The VTM that was urgently required needed to support viral replication, as well as other routine diagnostic approaches. We describe the preparation and validation testing of VTM for rapidly expanding diagnostic testing, where the capacity of the VTM to preserve viral integrity, for culture, isolation and full sequence analysis, was maintained.MethodsVTM was prepared using different methods of sterilization then ‘spiked’ with virus. The VTM was investigated using viral culture in Vero cells, and for nucleic acid detection by quantitative PCR (qPCR).ResultsThe best results were obtained by filter and autoclave-based sterilization. The VTM proved robust for culture-based analyses provided the inoculated VTM was stored at 4oC, and tested within 48 hours. The filtered VTM also supported PCR-based diagnosis for at least 5 days when the mock inoculated VTM was held at room temperature (RT).DiscussionThe manual handling of VTM production, including filling and sterilization, was optimized. SARS-CoV-2 was spiked into VTM to assess different sterilization methods and measure the effects of storage time and temperature upon VTM performance. While most diagnostic protocols will not require replication competent virus, the use of high quality VTM will allow for the next phase of laboratory analysis in the COVID-19 pandemic, including drug and antibody susceptibility analysis of re-isolated SARS--CoV-2, and for the testing of vaccine escape mutants.


2020 ◽  
Vol 35 (1) ◽  
pp. 25-33 ◽  
Author(s):  
Magnus Heldin ◽  
Urban Wiklund

AbstractGroundwood pulping is a process that employs large machines, making them difficult to use in research. Lab scale grinders exist, but even though they are smaller, the sizes of the grinding stones or segments make them cumbersome to exchange and tailor. This study presents a method and an apparatus for investigating the detailed mechanisms and the energy requirements behind the fibre separation process. A well-defined grinding tool was used at three different temperatures to demonstrate that the equipment can differentiate levels of energy consumption and defibration rates, confirming the well-known fact that a higher temperature facilitates defibration. It is also shown how the equipment can be used to study the influence of grinding parameters, exemplified by the effect of temperature on the way fibres are separated and the character of the produced fibres. A key feature of the equipment is the use and evaluation of small grinding surfaces, more readily designed, produced, evaluated and studied. This reduces both the cost and time necessary for testing and evaluating. At the same time, a technique to produce well defined grinding surfaces was employed, which is necessary for repeatability and robust testing, not achievable with traditional grinding stones.


2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Julie McAuley ◽  
Claire Fraser ◽  
Elena Paraskeva ◽  
Elizabeth Trajcevska ◽  
Michelle Sait ◽  
...  

Abstract Introduction The sudden arrival of the COVID-19 pandemic placed significant stresses on supply chains including viral transport medium (VTM). The VTM that was urgently required needed to support viral replication, as well as other routine diagnostic approaches. We describe the preparation and validation testing of VTM for rapidly expanding diagnostic testing, where the capacity of the VTM to preserve viral integrity, for culture, isolation and full sequence analysis, was maintained. Methods VTM was prepared using different methods of sterilization then ‘spiked’ with virus. The VTM was investigated using viral culture in Vero cells, and for nucleic acid detection by quantitative PCR. Results The best results were obtained by filter and autoclave-based sterilization. The VTM proved robust for culture-based analyses provided the inoculated VTM was stored at 4 °C, and tested within 48 h. The filtered VTM also supported PCR-based diagnosis for at least 5 days when the mock inoculated VTM was held at room temperature. Discussion The manual handling of VTM production, including filling and sterilization, was optimized. SARS-CoV-2 was spiked into VTM to assess different sterilization methods and measure the effects of storage time and temperature upon VTM performance. While most diagnostic protocols will not require replication competent virus, the use of high quality VTM will allow for the next phase of laboratory analysis in the COVID-19 pandemic, including drug and antibody susceptibility analysis of re-isolated SARS-CoV-2, and for the testing of vaccine escape mutants.


Separations ◽  
2021 ◽  
Vol 8 (12) ◽  
pp. 229
Author(s):  
Germán Eduardo Dévora-Isiordia ◽  
Alejandra Ayala-Espinoza ◽  
Luis Alberto Lares-Rangel ◽  
María Isela Encinas-Guzmán ◽  
Reyna Guadalupe Sánchez-Duarte ◽  
...  

A high percentage of the agricultural wells in the state of Sonora are overexploited, thus generating a significant degree of saline intrusion and abandonment by nearby communities. In this paper, the effect of temperature on the final concentration of diluted water was evaluated with variations in voltage and input concentration in a batch electrodialysis reversal (EDR) process in order to find the optimal operating conditions, with an emphasis on reducing the energy consumption and cost of desalinated water. Thirty-six samples were prepared: eighteen samples of 2000 mg/L total dissolved solids (TDS) and eighteen samples of 5000 mg/L TDS; brackish well water of 639 mg/L TDS and synthetic salt were mixed to obtain these concentrations. Three different temperatures (25, 30, and 35 °C) and two different voltages (10 and 20 V) were tested for each sample after evaluating the limiting current density. The best salt removal occurred in the 20 V sets, with 18.34% higher removal for the 2000 mg/L TDS experiments and 25.05% for the 5000 mg/L experiments (average between the 25 to 35 °C tests). The temperature positively affected the EDR, especially in the experiments at 10 V, where increasing by 10 °C increased the efficiency by 10.83% and 24.69% for 2000 and 5000 mg/L TDS, respectively. The energy consumption was lower with increasing temperature (35 °C), as it decreased by 1.405% and 1.613% for the 2000 and 5000 mg/L TDS concentrations, respectively (average between the 10 and 20 V tests), thus decreasing the cost per m3 of water.


Author(s):  
Adrien Calame ◽  
Lena Mazza ◽  
Adriana Renzoni ◽  
Laurent Kaiser ◽  
Manuel Schibler

In the context of an unprecedented shortage of nasopharyngeal swabs (NPS) or sample transport media during the coronavirus disease 2019 (COVID-19) crisis, alternative methods for sample collection are needed. To address this need, we validated a cell culture medium as a viral transport medium, and compared the analytical sensitivity of SARS-CoV-2 real-time RT-PCR in nasal wash (NW), oropharyngeal swab (OPS) and NPS specimens. Both the clinical and analytical sensitivity were comparable in these three sample types. OPS and NW specimens may therefore represent suitable alternatives to NPS for SARS-CoV-2 detection.


Author(s):  
Adrien Calame ◽  
Léna Mazza ◽  
Adriana Renzoni ◽  
Laurent Kaiser ◽  
Manuel Schibler

Abstract In the context of an unprecedented shortage of nasopharyngeal swabs (NPS) or sample transport media during the coronavirus disease 2019 (COVID-19) crisis, alternative methods for sample collection are needed. To address this need, we validated a cell culture medium as a viral transport medium, and compared the analytical sensitivity of SARS-CoV-2 RT-PCR in nasal wash (NW), oropharyngeal swab (OPS), and NPS specimens. Both the clinical and analytical sensitivity were comparable in these three sample types. OPS and NW specimens may therefore represent suitable alternatives to NPS for SARS-CoV-2 detection.


Author(s):  
Smita Tyagi ◽  
Rajesh Kumar Agarwal ◽  
Manish Agarwal ◽  
Manika Kapoor ◽  
Sakshi Tiwari

Background: The objective of this study was to estimate the incidence of vertical transmission COVID-19 by RT-PCR.Methods: In this hospital based prospective study, all COVID-19 positive pregnant women admitted in COVID ward in Muzaffarnagar medical college and hospital from April 2020 to January 2021 were included. A detailed history and examination was done and all routine investigations were done as per protocol. Samples were taken from amniotic fluid during vaginal delivery or caesarean section and collected in viral transport medium. Sample were also collected as nasal and oropharyngeal swab from neonate immediately after birth and sent for COVID 19 RT-PCR.Results: Out of total 50 cases; 43 (86%) neonates were delivered via LSCS and 7 (14%) by normal delivery, out of these 41 (82%) neonates were normal; 4 (8%) were IUGR, 2 (4%) were IUD, 2 (4%) were pre-term and 1 (2%) neonatal death. All 50 amniotic fluid as well as nasal and oropharyngeal samples of all neonates were negative by RT-PCR.Conclusions: Low vertical transmission may be due to the fact that antibodies are produced by mother that crosses the placenta and saves the fetus or there are highly specific immunological mediators in the placenta that do not allow the infection to pass to the fetus.


Author(s):  
D. T. Gauld ◽  
J. E. G. Raymont

The respiratory rates of three species of planktonic copepods, Acartia clausi, Centropages hamatus and Temora longicornis, were measured at four different temperatures.The relationship between respiratory rate and temperature was found to be similar to that previously found for Calanus, although the slope of the curves differed in the different species.The observations on Centropages at 13 and 170 C. can be divided into two groups and it is suggested that the differences are due to the use of copepods from two different generations.The relationship between the respiratory rates and lengths of Acartia and Centropages agreed very well with that previously found for other species. That for Temora was rather different: the difference is probably due to the distinct difference in the shape of the body of Temora from those of the other species.The application of these measurements to estimates of the food requirements of the copepods is discussed.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Aiyan Guan ◽  
Inge Van Damme ◽  
Frank Devlieghere ◽  
Sarah Gabriël

AbstractAnisakidae, marine nematodes, are underrecognized fish-borne zoonotic parasites. Studies on factors that could trigger parasites to actively migrate out of the fish are very limited. The objective of this study was to assess the impact of different environmental conditions (temperature, CO2 and O2) on larval motility (in situ movement) and mobility (migration) in vitro. Larvae were collected by candling or enzymatic digestion from infected fish, identified morphologically and confirmed molecularly. Individual larvae were transferred to a semi-solid Phosphate Buffered Saline agar, and subjected to different temperatures (6 ℃, 12 ℃, 22 ℃, 37 ℃) at air conditions. Moreover, different combinations of CO2 and O2 with N2 as filler were tested, at both 6 °C and 12 °C. Video recordings of larvae were translated into scores for larval motility and mobility. Results showed that temperature had significant influence on larval movements, with the highest motility and mobility observed at 22 ℃ for Anisakis spp. larvae and 37 ℃ for Pseudoterranova spp. larvae. During the first 10 min, the median migration of Anisakis spp. larvae was 10 cm at 22 ℃, and the median migration of Pseudoterranova spp. larvae was 3 cm at 37 ℃. Larval mobility was not significantly different under the different CO2 or O2 conditions at 6 °C and 12 ℃. It was concluded that temperature significantly facilitated larval movement with the optimum temperature being different for Anisakis spp. and Pseudoterranova spp., while CO2 and O2 did not on the short term. This should be further validated in parasite-infected/spiked fish fillets.


Toxins ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 134
Author(s):  
Ana Isabel Galván ◽  
Alicia Rodríguez ◽  
Alberto Martín ◽  
Manuel Joaquín Serradilla ◽  
Ana Martínez-Dorado ◽  
...  

Dried fig is susceptible to infection by Aspergillus flavus, the major producer of the carcinogenic mycotoxins. This fruit may be contaminated by the fungus throughout the entire chain production, especially during natural sun-drying, post-harvest, industrial processing, storage, and fruit retailing. Correct management of such critical stages is necessary to prevent mould growth and mycotoxin accumulation, with temperature being one of the main factors associated with these problems. The effect of different temperatures (5, 16, 25, 30, and 37 °C) related to dried-fig processing on growth, one of the regulatory genes of aflatoxin pathway (aflR) and mycotoxin production by A. flavus, was assessed. Firstly, growth and aflatoxin production of 11 A. flavus strains were checked before selecting two strains (M30 and M144) for in-depth studies. Findings showed that there were enormous differences in aflatoxin amounts and related-gene expression between the two selected strains. Based on the results, mild temperatures, and changes in temperature during drying and storage of dried figs should be avoided. Drying should be conducted at temperatures >30 °C and close to 37 °C, while industry processing, storage, and retailing of dried figs are advisable to perform at refrigeration temperatures (<10 °C) to avoid mycotoxin production.


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