A fundamental tenet of scientific research is that published results are open to independent validation and refutation. Minimum data standards aid data providers, users, and publishers by providing a specification of what is required to unambiguously interpret experimental findings. Here, we present the Minimum Information about a Flow Cytometry Experiment (MIFlowCyt) standard, stating the minimum information required to report flow cytometry (FCM) experiments. We brought together a crossdisciplinary international collaborative group of bioinformaticians, computational statisticians, software developers, instrument manufacturers, and clinical and basic research scientists to develop the standard. The standard was subsequently vetted by the International Society for Advancement of Cytometry (ISAC) Data Standards Task Force, Standards Committee, membership, and Council. The MIFlowCyt standard includes recommendations about descriptions of the specimens and reagents included in the FCM experiment, the configuration of the instrument used to perform the assays, and the data processing approaches used to interpret the primary output data. MIFlowCyt has been adopted as a standard by ISAC, representing the FCM scientific community including scientists as well as software and hardware manufacturers. Adoption of MIFlowCyt by the scientific and publishing communities will facilitate third-party understanding and reuse of FCM data. ' 2008 International Society for Advancement of Cytometry Key termsimmunology; fluorescence-activated cell sorting; knowledge representation FLOW cytometry (FCM) systems have been available to investigators for over 30 years, and the field continues to advance at a rapid rate. FCM has been responsible for major progress in basic and clinical research by enabling the phenotypic and functional characterization of individual cells in a high-throughput manner. Advances in the technology now allow for automated, multiparametric analyses of thousands of samples per day (1). Each data set can consist of multidimensional descriptions of millions of individual cells, producing data similar in size and complexity to gene expression microarrays. Like the microarray field, the ability to collect FCM data is outpacing the computational means for data handling and analysis. Furthermore, the lack of reporting standardization limits collaboration, independent validation/refutation, and meta-analysis, and thus minimizes the value of the wealth
More than 20 Synechococcus and Cyanobium isolates were obtained from central European subalpine lakes and sequenced for their 16S rRNA gene and part of the phycocyanin operon (cpc), specifically the intergenic spacer (IGS) between cpcB and cpcA, and corresponding flanking regions (cpcBA-IGS). Maximum-likelihood analyses revealed the existence of at least six to seven clusters of nonmarine picocyanobacteria within the picophytoplankton clade and support the conjecture of global dispersal for some closely related picocyanobacterial genotypes.
In this review the analytical techniques for measuring microplastics in sediment have been evaluated.
The shedding of pathogens by infected humans enables the use of sewage monitoring to conduct wastewater-based epidemiology (WBE). Although most WBE studies use data from large sewage treatment plants, timely data from smaller catchments are needed for targeted public health action. Traditional sampling methods, like autosamplers or grab sampling, are not conducive to quick ad hoc deployments and high-resolution monitoring at these smaller scales. This study develops and validates a cheap and easily deployable passive sampler unit, made from readily available consumables, with relevance to the COVID-19 pandemic but with broader use for WBE. We provide the first evidence that passive samplers can be used to detect SARS-CoV-2 in wastewater from populations with low prevalence of active COVID-19 infections (0.034 to 0.34 per 10,000), demonstrating their ability for early detection of infections at three different scales (lot, suburb, and city). A side by side evaluation of passive samplers ( n = 245) and traditionally collected wastewater samples ( n = 183) verified that the passive samplers were sensitive at detecting SARS-CoV-2 in wastewater. On all 33 days where we directly compared traditional and passive sampling techniques, at least one passive sampler was positive when the average SARS-CoV-2 concentration in the wastewater equaled or exceeded the quantification limit of 1.8 gene copies per mL ( n = 7). Moreover, on 13 occasions where wastewater SARS-CoV-2 concentrations were less than 1.8 gene copies per mL, one or more passive samplers were positive. Finally, there was a statistically significant ( p < 0.001) positive relationship between the concentrations of SARS-CoV-2 in wastewater and the levels found on the passive samplers, indicating that with further evaluation, these devices could yield semi-quantitative results in the future. Passive samplers have the potential for wide use in WBE with attractive feasibility attributes of cost, ease of deployment at small-scale locations, and continuous sampling of the wastewater. Further research will focus on the optimization of laboratory methods including elution and extraction and continued parallel deployment and evaluations in a variety of settings to inform optimal use in wastewater surveillance.
Quantifying the emissions of per- and polyfluoroalkyl substances (PFAS) from Australian wastewater treatment plants (WWTP) is of high importance due to potential impacts on receiving aquatic ecosystems. The new Australian PFAS National Environmental Management Plan recommends 0.23 ng L −1 of PFOS as the guideline value for 99% species protection for aquatic systems. In this study, 21 PFAS from four classes were measured in WWTP solid and aqueous samples from 19 Australian WWTPs. The mean ∑ 21 PFAS was 110 ng L −1 (median: 80 ng L −1 ; range: 9.3–520 ng L −1 ) in aqueous samples and 34 ng g −1 dw (median: 12 ng g −1 dw; range: 2.0–130 ng g −1 dw) in WWTP solids. Similar to WWTPs worldwide, perfluorocarboxylic acids were generally higher in effluent, compared to influent. Partitioning to solids within WWTPs increased with increasing fluoroalkyl chain length from 0.05 to 1.22 log units. Many PFAS were highly correlated, and PCA analysis showed strong associations between two groups: odd chained PFCAs, PFHxA and PFSAs; and 6:2 FTS with daily inflow volume and the proportion of trade waste accepted by WWTPs (as % of typical dry inflow). The compounds PFPeA, PFHxA, PFHpA, PFOA, PFNA, and PFDA increased significantly between influent and final effluent. The compounds 6:2 FTS and 8:2 FTS were quantified and F–53B detected and reported in Australian WWTP matrices. The compound 6:2 FTS was an important contributor to PFAS emissions in the studied Australian WWTPs, supporting the need for future research on its sources (including precursor degradation), environmental fate and impact in Australian aquatic environments receiving WWTP effluent.
This is the first study using flow cytometry to characterize the population dynamics of freshwater autotrophic picoplankton (APP) over a full seasonal cycle, the goal of which was to accurately quantify and qualify the natural APP populations in relation to major environmental parameters. In particular, we wanted to test current assumptions about the seasonal succession of prokaryotic and eukaryotic picoplankton cells, including the relationship between solitary picocyanobacteria and microcolonies. Using flow cytometry, we were able to efficiently characterize the abundances of 4 lake APP assemblages in Lake Mondsee, including that of a solitary picocyanobacterial population exhibiting high 'side scatter' values. Such cells were not readily enumerated by epifluorescence microscopy. Unlike Lakes Constance and Maggiore, we found no evidence of a spring peak in solitary picocyanobacteria -we propose that the lack of a spring peak in Lake Mondsee was due to weak stratification in March-April and relatively deep vertical mixing. Since summer declines in the abundance of solitary picocyanobacteria were associated with extended periods of reduced light availability, it is likely that such declines were in part due to low relative growth rates. Finally, we argue that the formation of microcolonies by picocyanobacteria is unlikely to be a strategy for more efficient nutrient recycling (e.g. Stockner & Antia 1986). Rather, we suggest that microcolonies reach high concentrations in surface and near-surface waters due to the production of a photosynthate-rich mucilage resulting from active photosynthesis during periods of severe nutrient deficiency.
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