The investigation of microplastics (MPs) in freshwater has received increased attention within the last decade. To date, sampling is mainly conducted at the surface of both rivers and lakes and only a few studies assessed the vertical distribution of MPs in the water column of freshwater bodies. To contribute to the understanding of MP pollution in the water column of freshwater lakes, this study evaluated the vertical profile of MPs in Lake Tollense considering particles between 63 and 5000 μm in size. Sampling was conducted on three occasions at three depths (surface, 7 m and 10 m) along a transect including eight sampling stations. The retrieved samples were digested with hydrogen peroxide and sodium hypochlorite and investigated via Nile Red staining and fluorescence microscopy. Subsequently, a sub-sample of stained particles was verified by μRaman-spectroscopy. The vertical distribution of MPs in Lake Tollense differed considerably between particle shapes (irregular particles (IPs) and fibers). Fibers did not show a noticeable pattern with depth and ranged between 22 fibers m⁻³ at 0 m to 19 fibers m⁻³ at 10 m. In contrast, IPs were distinctly less abundant in sub-surface samples with concentrations between 50 IPs m⁻³ at 0 m to 29 IPs m⁻³ at 10 m. Concerning IPs, buoyant polymers (mainly PE and PP) and concerning fibers PET and PP dominated the polymeric composition. Besides particle inherent properties, wind-induced mixing is likely affecting the intensity of vertical concentration gradients. This study highlights the need for depth-integrated sampling approaches in order to achieve representative data without over- or underestimating the overall abundances.

Since only a few studies have investigated effects of microplastics (MPs) by routes other than ingestion, this study was designed to analyze the accumulation patterns and transfer of toxic substances associated with microplastic exposure by simple attachment to (1) adult zebrafish (Danio rerio) gills and (2) zebrafish embryos. Two sizes of fluorescently labelled polymers (1–5 and 10–20 μm) loaded with the model polycyclic aromatic hydrocarbon (PAH) benzo[a]pyrene (BaP) were used to analyze fate, accumulation and transfer of microplastic-associated persistent organic pollutants (POPs) on gills and embryos.Results indicate that microplastics did not permanently accumulate at high amounts in adult zebrafish gills after 6 nor 24 h of incubation: Most particles only superficially adhered to the mucus layer on the filaments, which is constantly being excreted. In contrast, the smaller and heavier MPs (1–5 μm) accumulated in high numbers on the surface of zebrafish egg chorions. In both exposure scenarios, transfer of BaP could be visualized with fluorescence microscopy: A prominent BaP signal was visible both in gill filaments and arches after 6 and 24 h incubation and in zebrafish embryos after exposure to BaP-spiked microplastics. Furthermore, the gill EROD (Ethoxyresorufin-O-deethylase) assay showed a clear trend to CYP 1A (Cytochrom P450 1 A) induction via exposure to BaP-spiked microplastics. However, BaP from spiked microplastics did not reach sufficiently high concentrations to be able to induce morphological effects in the fish embryo toxicity test (FET). In contrast, control exposure to waterborne BaP did induce effects in the FET.As a conclusion, microplastics can also transfer POPs not only via ingestion, but also by simple attachment to epithelia or via the water column. However, further studies are needed to clarify if these interactions are of environmental concern relative to waterborne exposure to toxic substances.

Many reliable studies have provided evidence of microplastic ingestion by soil organisms. However, further research is required to determine the edible size of microplastics, especially given the ubiquity of microplastics and their adverse effects on the soil environment. Determining the size range of microplastics that can be ingested by soil organisms is crucial for the prediction of the exposure route and toxicity mechanisms of microplastics in soil. Springtails, organisms prevalent in a wide variety of soil ecosystems, can ingest or transport microplastics; however, direct evidence for this has not been reported. To address this knowledge gap, we designed dietary exposure experiments under laboratory conditions, using the springtail species Folsomia candida. The springtails were administered polyethylene microplastics in three different sizes (2, 34, and 66 μm) via their food for a short period of time; we further observed the intestinal presence of microplastics via fluorescence microscopy to determine the maximum edible size. We evaluated the effects of ingested microplastics on springtails by quantifying their moving behavior. The results show that the edible size of microplastics is < 66.0 ± 10.9 μm, and microplastics smaller than this can significantly reduce the velocity and distance of springtail movement by 74% ± 38% compared with the control group. Based on this finding, the broader fate and toxicity of microplastics in soil environments can be estimated. Furthermore, the average velocity and distance of springtail movement decreases in response to microplastic ingestion, highlighting the negative effects of microplastics on soil organisms.

Due to the potential toxicity of polycyclic aromatic hydrocarbons (PAHs) to humans, the uptake and translocation of PAHs in food crops have gained much attention. However, it is still unclear whether phloem participates in the acropetal translocation of PAHs in plants. Herein, the evidence for acropetal translocation of phenanthrene (a model PAH) via phloem is firstly tested. Wheat (Triticum aestivum L.) new leaves contain significantly higher phenanthrene concentration than old leaves (P < 0.05), and the inhibitory effect on phenanthrene translocation is stronger in old leaves after abscisic acid and polyvinyl alcohol (two common transpiration inhibitors) application. Phenanthrene concentration in xylem sap is slightly higher than in phloem sap. Ring-girdling treatment can significantly reduce phenanthrene concentration in castor bean (Ricinus communis L.) leaves. Two-photon fluorescence microscope images indicate a xylem-to-phloem and acropetal phloem translocation of phenanthrene in castor bean stem. Therefore, phloem is involved in the acropetal translocation of phenanthrene in wheat seedlings, especially when the xylem is not mature enough in scattered vascular bundle plants. Our results provide a deeper understanding of PAH translocation in plants, which have significant implications for food safety and phytoremediation enhancement of PAH-contaminated soil and water.

Terrestrial environments are subject to extensive pollution by plastics and, based on the slow degradation of plastics, are likely to act as long term sinks for microplastic debris. Currently the hazards of microplastics in soil and the potential impacts on soil organisms is poorly understood. Particularly the role of particle characteristics, such a size or polymer type, in dose-response relationships for microplastics is not known. The aim of this study was to assess the ingestion and toxicity of nylon (polyamide) particles, in three different size ranges, to Enchytraeus crypticus in a soil exposure. Effects were also compared with those of polyvinyl chloride (PVC) particles, in a single size range. Nylon particle ingestion was confirmed using fluorescence microscopy, with greatest ingestion for particles in the smallest size range (13–18 μm). To investigate how particle size affected survival and reproduction, E. crypticus were exposed to nylon particles in two well-defined size ranges (13–18 and 90–150 μm) and concentrations of 20, 50, 90 and 120 g/kg (2–12% w/w). An intermediate nylon size range (63–90 μm) and a larger sized PVC particle (106–150 μm), both at 90 g/kg, were also tested. Survival was not affected by either of the polymer types or sizes. Reproduction was significantly reduced, in a dose-dependent manner, by the nylon particles at high exposure concentrations (>90 g/kg). Smaller size ranges (13–18 μm) had a greater effect compared to larger size ranges (>63 μm), with a calculated EC₅₀ for the 13–18 μm size range of 108 ± 8.5 g/kg. This greater hazard could be qualitatively linked with the ingestion of a greater number of smaller particles. This study highlights the potential for toxic effects of plastics in small size ranges to soil organisms at high exposure concentrations, providing understanding of the hazards microplastics may pose in the terrestrial environment.

Protozoa as phagocytizing cells have been shown to integrate engineered nanoparticles (NPs), while the mechanism, dynamics and extent of such uptake are unclear. Here our fluorescence microscopy data showed that CdSe/ZnS quantum dots (QDs) with primary size of 12 nm were readily phagocytized into the food vacuoles of Tetrahymena thermophila in a time- and dose-dependent manner. Twenty hours after the exposure to QDs in sublethal concentration the clearance of the QDs from the cells was incomplete suggesting that phagocytosis of QDs into food vacuoles was not the only pathway of uptake by T. thermophila. This was further proven by the results that the inhibition of phagocytosis did not block the internalization of QDs into protozoans. This study provides a new insight into uptake and cellular trafficking of subtoxic concentrations of nanoparticles that may, due to prolonged retention times in the cells, pose risks by potentially becoming available to higher trophic levels.

Pesticides aid in crop-protection against pests and increase yield. However, the xenobiotic stress exerted by pesticides leads to the deterioration of human and animal health. There is a lacuna in our knowledge about their impact on the ocular surface The present work sheds light on this gap by analysing the deterioration of visual acuity as a consequence of pesticide induced xenobiotic stress and Notch pathway dysregulation.Alteration in the expression of vital components of the notch signalling was analyzed along the visual pathway with special focus on its two terminals-the cornea and the visual cortex, by mimicking the on-field scenario regarding chronic pesticide exposure in experimental murine model (Swiss albino mice; Mus musculus). Various aspects were taken into consideration through visual acuity tests, histological evaluations, culture analyses, wound healing assays, flowcytometric evaluation, fluorescence microscopic studies etc. Complete dysregulation of key players of the Notch signalling pathway was observed in both: cells of the ocular surface as well as those in the murine visual cortex post pesticide exposure, indicating activities relating to cell proliferation, differentiation and wound healing in the pesticide exposed samples. Ultra-microscopic analyses corroborated our findings by revealing the loss of fine neural processes in the visual cortex of the pesticide exposed murine samples, thereby hinting at delayed perception to visual stimuli. In vivo evaluations of the functional capacity of the neuroanatomical structures along the visual pathway also confirmed that pesticide exposure leads to severe damage along the various parts of the visual pathway, right from the ocular surface to the visual cortex.

Understanding of cadmium (Cd) uptake mechanism and development of lower Cd crop genotypes are crucial for combating its phytotoxicity and meeting 70% increase in food demand by 2050. Bio-accumulation of Cd continuously challenges quality of life specifically in regions without adequate environmental planning. Here, we investigated the mechanisms operating in Cd tolerance of two rice genotypes (Heizhan-43 and Yinni-801). Damage to chlorophyll contents and PSII, histochemical staining and quantification of reactive oxygen species (ROS), cell viability and osmolyte accumulation were studied to decipher the interactions between Cd and zinc (Zn) by applying two Cd and two Zn levels (alone as well as combined). Cd²⁺ and Ca²⁺ fluxes were also measured by employing sole Cd₁₀₀ (100 μmol L⁻¹) and Zn₅₀ (50 μmol L⁻¹), and their combination with microelectrode ion flux estimation (MIFE) technique. Cd toxicity substantially reduced chlorophyll contents and maximal photochemical efficiency (Fᵥ/Fₘ) compared to control plants. Zn supplementation reverted the Cd-induced toxicity by augmenting osmoprotectants and interfering with ROS homeostasis under combined treatments, particularly in Yinni-801 genotype. Fluorescence microscopy indicated a unique pattern of live and dead root cells, depicting more damage with Cd₁₀, Cd₁₅ and Cd₁₅+Zn₅₀. Our results confer that Cd²⁺ impairs the uptake of Ca²⁺ whereas, Zn not only competes with Cd²⁺ but also Ca²⁺, thereby modifying ion homeostasis in rice plants. This study suggests that exogenous application of Zn is beneficial for rice plants in ameliorating Cd toxicity in a genotype and dose dependent manner by minimizing ROS generation and suppressing collective oxidative damage. The observations confer that Yinni-801 performed better than Heizhan-43 genotype mainly under combined Zn treatments with low-Cd, presenting Zn fortification as a solution to increase rice production.

In recent years, reports of plastic debris in the gastrointestinal (GI) tract of fish have been well documented in the scientific literature. This, in turn, increased concerns regarding human health exposure to microplastics through the consumption of contaminated fish. Most of the available research regarding microplastic toxicity has focused on marine organisms through direct feeding or waterborne exposures at the individual level. However, little is known about the trophic transfer of microplastics through the aquatic food chain. Freshwater zooplankton Daphnia magna (hereafter Daphnia), and the fathead minnow Pimephales promelas (FHM), are well-known model species used in standard toxicological studies and ecological risk assessments that provide a simple model for trophic transfer. The aim of this study was to assess the tissue translocation, trophic transfer, and depuration of two concentrations (20 and 2000-part ml⁻¹) of 6 μm polystyrene (PS) microplastics particles between Daphnia and FHM. Bioconcentration factors (BCF) and bioaccumulation factors (BAF) were determined. Fluorescent microscopy was used to determine the number of particles in the water media and within the organs of both species. Throughout the five days of exposure, PS particles were only found within the GI tract of both species. The BCF for Daphnia was 0.034 ± 0.005 for the low concentration and 0.026 ± 0.006 for the high concentration. The BAF for FHM was 0.094 ± 0.037 for the low concentration and 0.205 ± 0.051 for the high concentration. Between 72 and 96 h after exposure all microplastic particles were depurated from both species. The presence of food had a significant effect on the depuration of microplastic particles from Daphnia but not for FHM. Based on the low BCF and BAF values for both species, rapid depuration rates, and null translocation of microplastic particles to organs and tissues from the GI tract, there is a low probability that microplastics will bioconcentrate and bioaccumulate under environmental conditions.

The severe and pervasive effects of multispecies foodborne microbial biofilms highlight the importance of rapid detection and diagnosis of contamination risk in the field using epifluorescence-based techniques (EBT) combined with automatic image-counting software. This study screened the hygiene quality of the environment, the carcass and the slaughtering equipment in the El-Kharga abattoir, New Valley Province, Egypt, to assess possible contamination during slaughter process. In addition, biofilm was assessed, and bacteria was enumerated by epifluorescence microscopy. Using both conventional and EBT, the highest bacterial counts were observed for the slaughtering equipment (6.6 and 5.2 cfu/cm2, respectively), followed by different parts of the carcass (4.1 and 4.4 cfu/cm2, respectively) and environmental samples (3.9 and 4.1 cfu/cm2, respectively). A high prevalence of E. coli O157:H7 was observed on the slaughtering equipment (25%), which also led to carcass (1%) contamination. Moreover, Enterobacteriaceae members were detected during examination, such as Klebsiella pneumoniae, Enterobacter aerogenes, and Raoultella ornithinolytica. Despite the relatively good hygiene quality of the abattoir environment, there is also a high risk associated with biofilm formation by pathogenic microorganisms on the slaughtering equipment. Moreover, EBT showed different structures of the biofilm, including those formed at different maturation stages, such as voids, microbubbles, channels and mushroom shapes. (EBT) microscopy combined with image-counting software could be a candidate substitute to estimate efficiently, precisely and rapidly the microbial aggregation and exposure risk in field than the conventional counting techniques.