Second round of an interlaboratory comparison of SARS-CoV2 molecular detection assays used by 45 veterinary diagnostic laboratories in the United States

The COVID-19 pandemic presents a continued public health challenge. Veterinary diagnostic laboratories in the United States use RT-rtPCR for animal testing, and many laboratories are certified for testing human samples; hence, ensuring that laboratories have sensitive and specific SARS-CoV2 testing methods is a critical component of the pandemic response. In 2020, the FDA Veterinary Laboratory Investigation and Response Network (Vet-LIRN) led an interlaboratory comparison (ILC1) to help laboratories evaluate their existing RT-rtPCR methods for detecting SARS-CoV2. All participating laboratories were able to detect the viral RNA spiked in buffer and PrimeStore molecular transport medium (MTM). With ILC2, Vet-LIRN extended ILC1 by evaluating analytical sensitivity and specificity of the methods used by participating laboratories to detect 3 SARS-CoV2 variants (B.1; B.1.1.7 [Alpha]; B.1.351 [Beta]) at various copy levels. We analyzed 57 sets of results from 45 laboratories qualitatively and quantitatively according to the principles of ISO 16140-2:2016. More than 95% of analysts detected the SARS-CoV2 RNA in MTM at ≥500 copies for all 3 variants. In addition, for nucleocapsid markers N1 and N2, 81% and 92% of the analysts detected ≤20 copies in the assays, respectively. The analytical specificity of the evaluated methods was >99%. Participating laboratories were able to assess their current method performance, identify possible limitations, and recognize method strengths as part of a continuous learning environment to support the critical need for the reliable diagnosis of COVID-19 in potentially infected animals and humans.     

Continuous Production of Biogenic Magnetite Nanoparticles by the Marine Bacterium Magnetovibrio blakemorei Strain MV-1T with a Nitrous Oxide Injection Strategy

Magnetotactic bacteria (MTB) produce magnetosomes, which are membrane-embedded magnetic nanoparticles. Despite their technological applicability, the production of magnetite magnetosomes depends on the cultivation of MTB, which results in low yields. Thus, strategies for the large-scale cultivation of MTB need to be improved. Here, we describe a new approach for bioreactor cultivation of Magnetovibrio blakemorei strain MV-1^T. Firstly, a fed-batch with a supplementation of iron source and N₂O injection in 24-h pulses was established. After 120 h of cultivation, the production of magnetite reached 24.5 mg∙L⁻¹. The maximum productivity (16.8 mg∙L⁻¹∙day⁻¹) was reached between 48 and 72 h. However, the productivity and mean number of magnetosomes per cell decreased after 72 h. Therefore, continuous culture in the chemostat was established. In the continuous process, magnetite production and productivity were 27.1 mg∙L⁻¹ and 22.7 mg∙L⁻¹∙day⁻¹, respectively, at 120 h. This new approach prevented a decrease in magnetite production in comparison to the fed-batch strategy.     

May the Force (Field) Be with You: On the Importance of Conformational Searches in the Prediction of NMR Chemical Shifts

NMR data prediction is increasingly important in structure elucidation. The impact of force field selection was assessed, along with geometry and energy cutoffs. Based on the conclusions, we propose a new approach named mix-J-DP4, which provides a remarkable increase in the confidence level of complex stereochemical assignments—100% in our molecular test set—with a very modest increment in computational cost.     

Potentially Beneficial Effects on Healthy Aging by Supplementation of the EPA-Rich Microalgae Phaeodactylum tricornutum or Its Supernatant—A Randomized Controlled Pilot Trial in Elderly Individuals

Dietary supplements that promote healthy aging are mostly warranted in an aging society. Because of age-related risks, anti-inflammatory and anti-oxidative agents such as microalgae are potential candidates for intervention. In a randomized controlled trial, we tested Phaeodactylum tricornutum (PT), a microalgae rich in eicosapentaenoic acid (EPA), carotenoids, vitamins, and β-glucans, cultured in bioreactors. In this pilot trial, 19 healthy elderly received supplements for two weeks based on either the whole PT (A), the β-1,3-glucan-rich PT supernatant (SupB), the combination thereof (A+SupB), or a Comparator product (Comp). The primary outcome variable plasma interleukin-6 was reduced after treatment with A+SupB compared to the Comp group (p = 0.04). The mobility parameters 5 s sit-to-stand test (p = 0.04 in the A group) and by trend gait speed (p = 0.08 in the A+SupB diet) were improved compared to Comp. No treatment effects were observed for fatty acids, compared to Comp but omega-6 to -3 fatty acid ratio (p = 0.006) and arachidonic acid/EPA ratio (p = 0.006) were reduced within group A+SupB. Further, the SupB study product reduced faecal zonulin (p = 0.03) compared to the Comp. The data revealed an anti-inflammatory and potentially anti-oxidative effect of particular PT preparations, suggesting that they might be suitable for effects in healthy elderly.     

1-O-Alkylglycerol Ethers from the Marine Sponge Guitarra abbotti and Their Cytotoxic Activity

The cytotoxicity-bioassay-guided fractionation of the ethanol extract from the marine sponge Guitarra abbotti, whose 1-O-alkyl-sn-glycerol ethers (AGEs) have not been investigated so far, led to the isolation of a complex lipid fraction containing, along with previously known compounds, six new lipids of the AGE type. The composition of the AGE fraction as well as the structures of 6 new and 22 previously known compounds were established using ¹H and ¹³C NMR, GC/MS, and chemical conversion methods. The new AGEs were identified as: 1-O-(Z-docos-15-enyl)-sn-glycerol (1), 1-O-(Z-docos-17-enyl)-sn-glycerol (2), 1-O-(Z-tricos-15-enyl)-sn-glycerol (3), 1-O-(Z-tricos-16-enyl)-sn-glycerol (4), 1-O-(Z-tricos-17-enyl)-sn-glycerol (5), and 1-O-(Z-tetracos-15-enyl)-sn-glycerol (6). The isolated AGEs show weak cytotoxic activity in THP-1, HL-60, HeLa, DLD-1, SNU C4, SK-MEL-28, and MDA-MB-231 human cancer cells. A further cytotoxicity analysis in JB6 P⁺ Cl41 cells bearing mutated MAP kinase genes revealed that ERK2 and JNK1 play a cytoprotective role in the cellular response to the AGE-induced cytotoxic effects.     

工厂化生产杏鲍菇细菌性病害的防治

工厂化生产杏鲍菇细菌性病害,是由假单孢杆菌引发的重要病害,在特定的控温、控湿、控光、控CO2条件下,每年重复利用出菇房18次以上的出菇房很容易发生、并迅速传播,给杏鲍菇工厂化生产企业带来不同程度的损失;研究、制定的10条预防措施,可以有效控制病害的发生、蔓延。     

粗甘油对生长猪生长性能、胴体性状及肉品的影响

通过一个为期138d的饲养试验来测定日粮中粗甘油对生长猪的生长性能,胴体性状,肉质指标和组织结构的影响。本试验使用的粗甘油包含84．51％甘油、11．95％水、2．91％氯化钠和0．32％甲醇。试验选用断奶8天后的仔猪共96头（48头公猪和48头母猪,平均体重为7．9±0．4kg）,按试验开始时的性别和体重分配到24个栏中（4头／栏）。日粮处理为玉米-豆粕型,     

Spatial and temporal analysis of coffee wilt disease caused by Fusarium xylarioides in Coffea canephora

Coffee wilt disease (CWD) caused by Fusarium xylarioides, considered to be a soil-inhabiting fungus, is endemic in several African countries, affecting commercially important coffee species and causing serious economic losses. Coffee wilt disease development in naturally infected Coffea canephora fields at the Coffee Research Institute in Uganda was assessed from April 2001 to March 2006 to generate information about temporal and spatial spread of the disease. Maps of diseased trees were also generated from the data. Semi-variance analysis was performed on the data to show the spatio-temporal structure of disease. Host influence on the spatio-temporal structure was deduced from the distribution pattern of diseased and healthy trees and analysis of variance. Results show that the temporal disease epidemic progress was slow. The disease was found to spread from initial infections to healthy neighbouring trees, resulting in an aggregated pattern. An infected tree could infect up to three healthy trees away, in any direction. Disease foci formed and expanded with time, coalescing but punctuated in spots planted with resistant hosts. There were varying levels of susceptibility among host genotypes, affecting the rates and levels of epidemic development. The implications of the findings to the control of CWD are discussed.