Nickel nitrate is a heavy metal known as an environmental contaminant due to its toxicity, long environmental half-lives, and capacity for bioaccumulation.  This study aims to determine chromosomal aberration, nuclear alteration, and cell death in Allium cepa var. aggregatum L. root caused by different nickel concentrations.  Roots of Allium cepa var. aggregatum were induced by soaking bulbs in water, then transferred to a solution containing nickel (Ni) at a concentration of 20 ppm, 30 ppm, and 40 ppm for 72 hours.  Root tip mitotic chromosome preparations were done by the squash method.  The chromosome was stained with aceto-orcein and chromosomal damages were observed under a microscope.  The results showed that the mitotic index decreased from 5.025% at control to 3.144%, 2.467%, and 2.181% at immersion with 20 ppm, 30 ppm 40 ppm nickel nitrate, respectively.  Anaphase and telophase indexes in roots with Ni treatments were lower than in control, suggesting that nickel inhibits cell division.  Nickel nitrate induced chromosomal damages and nuclear abnormalities, such as sticky chromosome, fragmented chromosome, chromosome bridge and chromosome laggard, micronuclei, binucleate and nuclear budding.  The percentage of chromosomal damage increases with a higher concentration of Ni.  In situ cell visualization showed that the higher the nickel concentration, the more coloured the root tips indicating high levels of cell death.

In this study, the purpose was to investigate the histopathological, genotoxic effect, oxidative stress and cell death due to Metronidazole (MTZ), which is a 5-nitroimidazole compound used widely for the treatment of anaerobic organism infections in fish and humans on gill and liver tissues of Oncorhynchus mykiss.Trout fishes were exposed to 5, 10, and 20 mg/L of MTZ in the aquariums for 2, 4 and 8 days. Staining technics namely H&E, NOS immunohistochemistry, and TUNEL were performed to determine histopathological changes, oxidative damage and apoptosis. Additionally, smear preparations were also prepared from gill blood for genotoxic evaluations. The organ damage started in the 2nd day with 5 mg/L MTZ application and effects increased per duration and dose-dependent manner. It was observed that the gills had the primary and secondary lamellae lengths, with formation of clavate lamellae, fusion in secondary lamellae, separation of epithelium and aneurysm. Regional necrosis, vacuolization of hepatocytes, pycnotic nucleus, enlarged sinusoids were also determined in the liver. NOS immunoreactivity increased with the inducible immunoreactivity (iNOS) that was more prominent when compared to the endothelial immunoreactivity (eNOS). Apoptotic immunoreactivity was higher in the 10 mg 8th day experimental group at liver and gills, and was lower 20 mg 8th day experimental group. When the gills and liver compared with each other, in all doses, immunoreactivity was lower in gills, compared with liver. Genotoxic examinations showed that both number of micronucleated erythrocytes and nuclei abnormalities were higher in MTZ-treated groups.

peer reviewed | Metconazole (MEZ), a chiral triazole fungicide, produces enantioselective adverse effects in non-target organisms. Among MEZ's isomers, cis-MEZ displays robust antimicrobial properties. Evaluating MEZ and cis-MEZ's toxicity may mitigate fungicide usage and safeguard non-target organisms. Our study evaluated the toxicity of MEZ and its cis-isomers at concentrations of 0.02, 0.2, 2, and 4 mg L−1. We report stereoselectivity and severe cardiovascular defects in zebrafish, including pericardial oedema, decreased heart rate, increased sinus venous and bulbous arteries distances, intersegmental vessel defects, and altered cardiovascular development genes (hand2, gata4, nkx2.5, tbx5, vmhc, amhc, dll4, vegfaa, and vegfc). Further, MEZ significantly increased oxidative stress and apoptosis in zebrafish, primarily in the cardiac region. Isoquercetin, an antioxidant found in plants, partially mitigates MEZ-induced cardiac defects. Furthermore, MEZ upregulated the Wnt/β-catenin pathway genes (wnt3, β-catenin, axin2, and gsk-3β) and β-catenin protein expression. Inhibitor of Wnt Response-1 (IWR-1) rescued MEZ-induced cardiotoxicity. Our findings highlight oxidative stress, altered cardiovascular development genes, and upregulated Wnt/β-catenin signaling as contributors to cardiovascular toxicity in response to MEZ and cis-MEZ treatments. Importantly, 1R,5S-MEZ exhibited greater cardiotoxicity than 1S,5R-MEZ. Thus, our study provides a comprehensive understanding of cis-MEZ's cardiovascular toxicity in aquatic life. © 2024 Elsevier Ltd

Nowadays, the ubiquitous distribution and increasing abundance of P⁺ᴵᴵᴵ in waterbodies have caused serious concerns regarding its bioavailability and potential toxicity. However, our knowledge on these issues is relatively limited. We addressed previously unknown effects of P⁺ᴵᴵᴵ on three dominate algae species i.e. Microcystic aeruginosa (M. aeruginosa), Chlorella pyrenoidesa (C. pyrenoidesa) and Cyclotella. sp in eutrophic waterbodies in China. Remarkable declines in biomass, specific growth rate and Chl-a of algae cells treated with 0.01–0.7 mg/L P⁺ᴵᴵᴵ as sole or an alternative P source were observed, indicating P⁺ᴵᴵᴵ had an inhibitory effect on the algal growth. Besides, the intracellular enzyme activities e.g superoxide dismutase (SOD) and malondialdehyde (MDA) were significantly increased with P⁺ᴵᴵᴵ stress. M. aeruginosa and Cyclotella. sp cells seemed to be more sensitive to P⁺ᴵᴵᴵ toxicity than C. pyrenoidesa since cell membrane suffered more serious stress and destruction. These findings combined, it confirmed P⁺ᴵᴵᴵ could not be utilized as bioavailable P, but had certain toxicity to the tested algae. It indicated that the increased P⁺ᴵᴵᴵ abundance in eutrophic waterbodies would accelerate the algal cell death, which could have a positive effect against algal blooms. Our results provide new insights into assessing the ecological risks of P⁺ᴵᴵᴵ in aquatic environments.

The mosquito Aedes aegypti is a primary vector for major arboviruses, and its control is mainly based on the use of insecticides. Caffeine and spent coffee grounds (CG) are potential agents in controlling Ae. aegypti by reducing survival and blocking larval development. In this study, we analyzed the effects of treatment with common CG (CCG: with caffeine), decaffeinated CG (DCG: with low caffeine), and pure caffeine on the survival, behavior, and morphology of the midgut of Ae. aegypti under laboratory conditions. Third instar larvae (L3) were exposed to different concentrations of CCG, DCG, and caffeine. All compounds significantly affected larval survival, and sublethal concentrations reduced larval locomotor activity, delayed development, and reduced adult life span. Damage to the midgut of treated larvae included changes in epithelial morphology, increased number of peroxidase-positive cells (more abundant in DCG-treated larvae), and caspase 3-positive cells (more abundant in CCG-treated larvae), suggesting that the treatments triggered cell damage, leading to activation of cell death. In addition, the treatments reduced the FMRFamide-positive enteroendocrine cells and dividing cells compared to the control. CG and caffeine have larvicidal effects on Ae. aegypti that warrant field testing for their potential to control mosquitoes.

Cigarette smoke (CS) affects immune functions, leading to severe outcomes in smokers. Robust evidence addresses the immunotoxic effects of combustible tobacco products. As heat-not-burn tobacco products (HNBT) vaporize lower levels of combustible products, we here compared the effects of cigarette smoke (CS) and HNBT vapor on Jurkat T cells. Cells were exposed to air, conventional cigarettes or heatsticks of HNBT for 30 min and were stimulated or not with phorbol myristate acetate (PMA). Cell viability, proliferation, reactive oxygen species (ROS) production, 8-OHdG, MAP-kinases and nuclear factor κB (NFκB) activation and metallothionein expression (MTs) were assessed by flow cytometry; nitric oxide (NO) and cytokine levels were measured by Griess reaction and ELISA, respectively. Levels of metals in the exposure chambers were quantified by inductively coupled plasma mass spectrometry. MT expressions were quantified by immunohistochemistry in the lungs and liver of C57Bl/6 mice exposed to CS, HNBT or air (1 h, twice a day for five days: via inhalation). While both CS and HBNT exposures increased cell death, CS led to a higher number of necrotic cells, increased the production of ROS, NO, inflammatory cytokines and MTs when compared to HNBT-exposed cells, and led to a higher expression of MTs in mice. CS released higher amounts of metals. CS and HNBT exposures decreased PMA-induced interleukin-2 (IL-2) secretion and impaired Jurkat proliferation, effects also seen in cells exposed to nicotine. Although HNBT vapor does not activate T cells as CS does, exposure to both HNBT and CS suppressed proliferation and IL-2 release, a pivotal cytokine involved with T cell proliferation and tolerance, and this effect may be related to nicotine content in both products.

The frequent exposure of bees to a wide variety of fungicides, on crops where they forage, can be considered a stressor factor for these pollinators. The organisms are exposed both to the fungicide active ingredients and to the adjuvants of commercial formulations. All these ingredients are brought to the hive by bee foragers through contaminated pollen and nectar, thus exposing also immature individuals during larval phase. This work aimed to compare the effects of larval exposure to the fungicide pyraclostrobin (active ingredient and commercial formulation) and its influence on the cytotoxicity to midguts in adults, which were inoculated with the Nosema ceranae spores in the post-emergence stage. Under laboratory conditions, Apis mellifera larvae received an artificial diet containing fungicide solution from the third to the sixth day of the feeding phase. One-day-old adult workers ingested 100,000 infectious N. ceranae spores mixed in sucrose solution. Effects on midgut were evaluated through cellular biomarkers of stress and cell death. The exposure to the fungicide (active ingredient and commercial formulation) did not affect the larval post-embryonic development and survival of adult bees. However, this exposure induced cytotoxicity in the cells of the midgut, showed by the increase in DNA fragmentation and alteration in the HSP70 immunolabeling pattern. Without the pathogen, the midgut cytotoxic effects and HSP70 immunolabeling of the organisms exposed to the commercial formulation were lower when compared to the exposure to its active ingredient. However, in the presence of the pathogen, the cytotoxic effects of the commercial formulation to the adult bees’ midgut were potentialized. The pathogen N. ceranae increased the damage to the intestinal epithelium of adult bees. Thus, realistic doses of pyraclostrobin present in beebread consumed by larvae can affect the health and induce physiological implications to the midgut functions of the adult bees.

Heavy metal tolerance of microorganisms is the basis of heavy metal removal by growing cells. In this study, a cross-protection effect generated by salt stress significantly enhanced the cadmium tolerance of multi-stress-tolerant Pichia kudriavzevii. Comparative transcriptome analysis using RNA-Seq linked with physiological and biochemical observation was used to elucidate the underlying mechanisms of the improved cadmium tolerance. The expression of cadmium transport related genes (GSTY2, GLR1, GLO2, YCF1 and YOR1), GSH content and GST activity were elevated by salt stress, suggesting enhanced cadmium conjugation and detoxification in yeast cells. The inhibited cadmium uptake by ZRT1 and enhanced cadmium efflux by YOR1 contributed to the decrease in the intracellular cadmium concentration. The improved expression of antioxidant enzyme genes (SOD1, SOD2, SOD6, CAT1 and PRXIID), along with the enhanced activities of antioxidant enzymes (SOD, CAT and POD) resulted in a decrease in cadmium-induced ROS production, protein carbonylation, lipid peroxidation and cell death. The abundant expression of heat shock protein genes (HSP12, HSP10 and SSC1) and genes related to trehalose synthesis (TPS1 and TSL1) induced by salt stress protected yeast cells against complex stress conditions, contributing to the improved cadmium tolerance. These findings will be useful to develop cadmium-tolerant yeasts for cadmium removal by growing cells.

An account of histo-cytological and ultrastructural studies on ozone effect on crop and forest species in Italy is given, with emphasis on induced cell death and the underlying mechanisms. Cell death phenomena possibly due to ambient O3 were recorded in crop and forest species. In contrast, visible O3 effects on Mediterranean vegetation are often unclear. Microscopy is thus suggested as an effective tool to validate and evaluate O3 injury to Mediterranean vegetation. A DAB-Evans blue staining was proposed to validate O3 symptoms at the microscopic level and for a pre-visual diagnosis of O3 injury. The method has been positively tested in some of the most important crop species, such as wheat, tomato, bean and onion and, with some restriction, in forest species, and it also allows one to gain some very useful insights into the mechanisms at the base of O3 sensitivity or tolerance. Ozone-induced cell death is a frequent phenomenon in Mediterranean conditions, not only in the most sensitive crops but also in forest species.

Graphene oxide (GO) has been widely studied and applied in numerous industrial fields and biomedical fields for its excellent physical and chemical properties. Along with the production and applications of GO persist increasing, the environmental health and safety risk (EHS) of GO has been widely studied. However, previous studies almost focused on the biotoxicity of pristine GO under a relatively high exposure dose, without considering its transformation process within environmental and biological mediums. Meanwhile, its secondary toxicity or synergistic effects have not been taken seriously. Here, two different kinds of artificial lung fluids were adopted to incubate pristine GO to mimic the biotransformation process of GO in the lung fluids. And, we explored that biotransformation within the artificial lung fluids could significantly change the physicochemical properties of GO and could enhance its biotoxicity. To reveal the synergistic effects of GO and toxic metal ions, we uncovered that GO could enhance the intracellular content of metal ions by inhibiting the efflux function of ATP binding cassette (ABC) transporters which are distributed on the cellular membrane, and artificial lung fluids incubation of GO could enhance this synergistic effect. Finally, toxic metal ions induced a series of toxic reactions through oxidative stress response and promoted cell death. Moreover, consistent with the results of in vitro experiments, the lungs of mice exposed to GOs combined with Cd exhibited significant inflammation and oxidative stress compared with Cd treatment alone, and it was more remarkable within the mice which were treated with bio-transformed GOs. In summary, this study explored the impact and mechanism of biotransformation of GO in the lung fluids on the synergistic and secondary effects between GO and metal ions.