Integrating time since deposition estimation of bloodstains into a DNA profiling workflow: A novel approach using fluorescence spectroscopy.

Determining the time since deposition (TSD) of bloodstains is important to establish a timeline of bloodshed, while DNA profiling addresses identity (source attribution). Traditionally treated as separate processes, this study integrates TSD estimation into routine DNA profiling by analyzing typically discarded cell lysate (eluates) from spin-column-based DNA extractions. Fluorescence spectroscopy was used to analyze eluates from bloodstains deposited up to 99 weeks.

Injection of Ortho-Functionalized Tetrafluorinated Azobenzene-Containing siRNAs into Japanese Medaka Embryos for Photocontrolled Gene Silencing.

This article describes the detailed methodology of how to inject photoswitchable ortho-functionalized tetrafluorinated short interfering RNAs (F-siRNAs) into a single cell of stage-two Japanese medaka (Oryzias latipes) embryos and how to control gene silencing with different wavelengths of light. Many of the prior papers describing Japanese medaka embryo injections omit key information. As such, this article aims to give an in-depth explanation as to how the NanoJect III microinjector can be used for this purpose.

Injection of Reversible Optically Controlled Short Interfering RNA into Japanese Medaka Embryos () to Regulate Gene Silencing.

Photoswitchable -functionalized tetrafluorinated azobenzene-modified siRNAs (F-azo-siRNAs) were synthesized using solid-phase phosphoramidite chemistry. The activity of an F-azo-siRNA targeting enhanced green fluorescence protein (eGFP) in transgenic (Tg) Japanese Medaka () was reversibly photocontrolled with blue (470 nm) and green (530 nm) light, to activate and inactivate the siRNA, respectively. This study highlights the first reversible study with photoswitchable siRNA.

Metabolic, neurotoxic and immunotoxic effects of PFAAs and their mixtures on the proteome of the head kidney and plasma from rainbow trout (Oncorhynchus mykiss).

PFAAs (Perfluoroalkyl acids) are a class of bioaccumulative, persistent and ubiquitous environmental contaminants which primarily occupy the hydrosphere and its sediments. Currently, a paucity of toxicological information exists for short chain PFAAs and complex mixtures. In order to address these knowledge gaps, we performed a 3-week, aqueous exposure of rainbow trout to 3 different concentrations of a PFAA mixture (50, 100 and 500 ng/L) modeled after the composition determined in Lake Ontario.

Waterborne exposure to the antineoplastic 5-fluorouracil alters lipid composition in larval zebrafish (Danio rerio).

Antineoplastic medications are present in aquatic environments and are measured at relatively high concentrations in hospital sewage effluent. Thus, it is important to characterize risk associated with waterborne exposures to anticancer drugs. The drug 5-fluorouracil (5-FU) is used to treat several types of cancers, acting to inhibit cell division and cellular metabolism.

An alternative method for monitoring and interpreting influenza A in communities using wastewater surveillance.

Seasonal influenza is an annual public health challenge that strains healthcare systems, yet population-level prevalence remains under-reported using standard clinical surveillance methods. Wastewater surveillance (WWS) of influenza A can allow for reliable flu surveillance within a community by leveraging existing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) WWS networks regardless of the sample type (primary sludge vs. primary influent) using an RT-qPCR-based viral RNA detection method for both targets.

Characterizing the origin of blood plasma proteins from organ tissues in rainbow trout (Oncorhynchus mykiss) using a comparative non-targeted proteomics approach.

Protein expression patterns adapt to various cues to meet the needs of an organism. The dynamicity of an organism's proteome can therefore reveal information about an organism's health. Proteome databases contain limited information regarding organisms outside of medicinal biology.

Proteome changes in an aquatic invertebrate consumer in response to different nutritional stressors.

Nutrient imbalances in zooplankton are caused by the differences in elemental content of producers and the demand for elements in consumers, which alter the life-history traits in consumers. Changes in life-history traits are mediated through metabolic pathways that affect gene expression and the metabolome. However, less is known about proteomic changes to elemental-limitation in zooplankton.

Non-Lethal Blood Sampling of Fish in the lab and Field With Methods for Dried Blood Plasma Spot Omic Analyses.

There is global acknowledgment that humane methods in animal research are a priority, but few environmental effects monitoring programs use nonlethal methods for fish. The goal of the present study was to determine the impacts of sampling small volumes of blood in larger-bodied fish on survival and healing. In addition to evaluating survival following blood sampling, we evaluated the utility of dried blood spots as an alternative for sample processing and storage in the field.

Detection of SARS-CoV-2 Proteins in Wastewater Samples by Mass Spectrometry.

The recent COVID-19 pandemic overwhelmed the health system worldwide, and there was a need to track outbreaks and try to use this information as an early warning system. Wastewater-based epidemiology (WBE) enabled detection of the SARS-CoV-2 virus in wastewater treatment plant influents. Until now, the most used technique for this detection has been the quantitative polymerase chain reaction (qPCR)-based quantification of SARS-CoV-2 RNA.

Fish tissue accumulation and proteomic response to microcystins is species-dependent.

Cyanotoxins including microcystins are increasing globally, escalating health risks to humans and wildlife. Freshwater fish can accumulate and retain microcystins in tissues; however, uptake and depuration studies thus far have not exposed fish to microcystins in its intracellular state (i.e.

Cyanotoxins within and Outside of Cause Adverse Effects in Rainbow Trout ().

The global expansion of toxic blooms, and production of cyanotoxins including microcystins, are an increasing risk to freshwater fish. Differentiating intracellular and extracellular microcystin toxicity pathways (i.e.

Hypoxic responses in Oncorhynchus mykiss involve angiogenesis, lipid, and lactate metabolism, which may be triggered by the cortisol stress response and epigenetic methylation.

The incidence of hypoxia in water bodies is increasing more rapidly than aquatic life can adapt. This study aimed to determine the effects of hypoxia on fish physiology, as well as protein expression through proteomics. To do this, 40 rainbow trout were divided into normoxic control (11.

Are we forgetting the "proteomics" in multi-omics ecotoxicology?

Proteomics plays a significant role in discerning the effects of chemical exposures in animal taxa. Multi-omics applications have become more pervasive in toxicology, however questions remain about whether proteomics is being utilized by the community to its full potential - are we placing too much stock in transcriptomics and other omics approaches for developing adverse outcome pathways? Proteins are more relevant than transcripts because they are direct mediators of the resulting phenotype. There is also rarely perfect stoichiometry between transcript and protein abundance and transcript abundance may not accurately predict physiologic response.

Brown bullhead at the St. Lawrence River (Cornwall) Area of Concern: health and endocrine status in the context of tissue concentrations of PCBs and mercury.

The St. Lawrence River, at Cornwall Ontario, has accumulated sediment contaminants, mainly mercury (Hg) and polychlorinated biphenyls (PCBs), from industrial point sources over many years. Although those sources are past, the river at Cornwall remains an Area of Concern (AOC).

Shotgun proteomics analysis reveals sub-lethal effects in Daphnia magna exposed to cell-bound microcystins produced by Microcystis aeruginosa.

Microcystins that are cell-bound within Microcystis have demonstrated the ability to cause lethal and reproductive impairment in Daphnia, who constitute an important part of aquatic food chains and are known to feed on viable cyanobacterial cells. Recent advances in environmental toxicogenomics can be used to better understand the mechanistic effects from exposure to cell-bound microcystins in Daphnia; however, there remains a need to examine the effects of microcystins exposure as a function of dose and time in order to help elucidate the progression of (sub-)lethal effects. This study examines the effects of cell-bound microcystin exposure in Daphnia magna as a function of dose and time with shotgun proteomics in order to measure and provide insightful evidence describing functional mechanisms from, and relationships between, protein populations in response to toxic Microcystis aeruginosa.

Toxicity of the aquatic herbicide, reward®, on the fathead minnow with pulsed-exposure proteomic profile.

The objectives of this study were to assess the lethal and sub-lethal effects of the aquatic herbicide commercial formulation, Reward® (373 g/L DB), using application scenarios prescribed by the manufacturer. Specifically, a 14 d period between applications of Reward® in a water body undergoing treatment is required, yet the effects of these 'pulse' exposure scenarios on aquatic wildlife such as fish are unknown. In the first experiment early life stage FHM were exposed to a continuous DB concentrations from 0.

Hepatic proteome network data in zebrafish () liver following dieldrin exposure.

Dieldrin is an environmental contaminant that adversely affects aquatic organisms. The data presented in this study are proteomic data collected in liver of zebrafish that were exposed to the pesticide in a dietary exposure. For label free proteomics, data were collected with a quadrupole Time-of-Flight mass spectrometer and for iTRAQ proteomics, data were acquired using a hybrid quadrupole Orbitrap (Q Exactive) MS system.

Label-free and iTRAQ proteomics analysis in the liver of zebrafish (Danio rerio) following dietary exposure to the organochlorine pesticide dieldrin.

The organochlorine dieldrin (DLD) bioaccumulates in lipid-rich tissues and is associated with immunosuppression, altered metabolism, and cancer. The objective of this study was to determine the effect of DLD on the hepatic proteome in zebrafish following dietary treatment as the liver is central to metabolism. Females were fed a control dose or one of three doses of DLD-contaminated food pellets over 21 days.

Reduced anxiety is associated with the accumulation of six serotonin reuptake inhibitors in wastewater treatment effluent exposed goldfish Carassius auratus.

Pharmaceuticals and personal care products (PPCPs) have been found in wastewater treatment plant (WWTP) effluents and their recipient watersheds. To assess the potential of WWTP effluents to alter fish behaviour, we caged male goldfish (Carassius auratus) for 21-days at three sites along a contamination gradient downstream from a WWTP which discharges into Cootes Paradise Marsh, on the western tip of Lake Ontario. We also included a fourth caging site as an external reference site within Lake Ontario at the Jordan Harbour Conservation Area.

Altered expression of metabolites and proteins in wild and caged fish exposed to wastewater effluents in situ.

Population growth has led to increased global discharges of wastewater. Contaminants that are not fully removed during wastewater treatment, such as pharmaceuticals and personal care products (PPCPs), may negatively affect aquatic ecosystems. PPCPs can bioaccumulate causing adverse health effects and behavioural changes in exposed fish.

Inhibition of immune responses and related proteins in Rhamdia quelen exposed to diclofenac.

Nonsteroidal anti-inflammatory drugs are among the most widely detected pharmaceuticals in surface water worldwide. The nonsteroidal anti-inflammatory drug diclofenac is used to treat many types of pain and inflammation. Diclofenac's potential to cause adverse effects in exposed wildlife is a growing concern.