The agricultural sector uses fertilizers such as urea to add more nutrients to the soil needed for plant growth. Although it is cost-effective in crop production, indiscriminate use of nitrate-based fertilizer may result in behavioural, morphological, and physiological alterations on non-target organisms. This study determined the angiogenesis activity in the chorioallantoic membrane of urea-exposed duck embryos. It also investigated the weight, morphometries, and liver histopathology to gather more information on urea fertilizer's toxicity. It was observed that urea promoted angiogenesis in the CAM of duck embryos, especially at higher concentrations (P<0.05). Embryos treated with urea resulted in an alteration of the head-beak length (P<0.05). However, weight, crown-rump length, forelimb length, and hind limb length were not affected. The developing liver of urea-treated embryos showed distortion of the central vein shape and had larger sinusoidal spaces. The presence of Kupffer cells and lipid droplets were observed in the treated section. Congestion of blood cells, haemorrhage, and necrosis of hepatocytes were also observed in the tissue suggesting the extent of damage caused by the fertilizer. The findings of this study showed multiple developmental effects of urea on duck embryos. Further investigations are needed to shed more light on the toxicity of urea fertilizer on vertebrates.

Microplastics (MPs), an emerging environmental pollutant, have been clarified to induce testicular disorder in mammals. And the current studies have delineated a correlation between gut microbiota and male reproduction. However, it's still unclear whether gut microbiota gets involved in MPs-induced reproductive toxicity. In this work, we constructed a mouse model drinking 5 μm polystyrene-MPs (PS-MPs) at the concentrations of 100 μg/L and 1000 μg/L for 90 days. Evident histological damage, spermatogenetic disorder and hormones synthesis inhibition were observed in PS-MPs exposed mice. With fecal microbiota transplantation (FMT) trial, the recipient mice exhibited gut microbial alteration, and the elevated abundance of Bacteroides and Prevotellaceae_UCG-001 were positively correlated with testicular disorder according to spearman correlation analysis. Mechanistically, increased proportion of pro-inflammatory bacteria may drive translocation of T helper 17 (Th17) cells, resulting in overproduced interleukin (IL)-17 A and downstream inflammatory response in both the mice exposed to PS-MPs and corresponding recipient mice. In summary, our findings revealed the critical role of gut microbiota in PS-MPs-induced reproductive toxicity, and tried to elucidate the underlying mechanism of gut microbial dysregulation-mediated IL-17 A signaling pathway. Furthermore, this study also provides the research basis for gut microbiota-targeted treatment of male infertility in the future.

Amongst other threats, the world’s oceans are faced with man-made pollution, including an increasing number of microparticulate pollutants. Sponges, aquatic filter-feeding animals, are able to incorporate fine foreign particles, and thus may be a potential bioindicator for microparticulate pollutants. To address this question, 15 coral reef demosponges sampled around Bangka Island (North Sulawesi, Indonesia) were analyzed for the nature of their foreign particle content using traditional histological methods, advanced light microscopy, and Raman spectroscopy. Sampled sponges accumulated and embedded the very fine sediment fraction (<200 μm), absent in the surrounding sand, in the ectosome (outer epithelia) and spongin fibers (skeletal elements), which was confirmed by two-photon microscopy. A total of 34 different particle types were identified, of which degraded man-made products, i.e., polystyrene, particulate cotton, titanium dioxide and blue-pigmented particles, were incorporated by eight specimens at concentrations between 91 and 612 particle/g dry sponge tissue. As sponges can weigh several hundreds of grams, we conservatively extrapolate that sponges can incorporate on average 10,000 microparticulate pollutants in their tissue. The uptake of particles, however, appears independent of the material, which suggests that the fluctuation in material ratios is due to the spatial variation of surrounding microparticles. Therefore, particle-bearing sponges have a strong potential to biomonitor microparticulate pollutants, such as microplastics and other degraded industrial products.

The use of chemotherapy agents has been growing worldwide, due to the increase number of cancer cases. In several countries, mainly in Europe countries, these drugs have been detected in hospitals and municipal wastewaters. In Brazil this issue is poorly explored. The main goal of this study was to assess the presence of three anti-cancer drugs, 5-fluorouracil (5-FU), gemcitabine (GEM) and cyclophosphamide (CP), and two metabolites, alpha-fluoro-beta-alanine (3-NH₂-F) and 2′-deoxy-2′,2′-difluorouridine (2-DOH-DiF), in effluents from a large cancer hospital, in the municipal wastewater treatment plant (WWTP) influent and effluent, and also to evaluate toxicity of the mixtures of these compounds by ecotoxicological testing in zebrafish. The sample collections were performed in Barretos Cancer Hospital of the large cancer center in Brazil. After each collection, the samples were filtered for subsequent Liquid Chromatography Mass Spectrometry analysis. The presence of CP, GEM, and both metabolites (3-NH₂-F and 2-DOH-DiF) were detected in the hospital wastewater and the WWTP influent. Three drugs, GEM, 2-DOH-DiF and CP, were detected in the WWTP effluent. Two drugs were detected below the limit of quantification, 2-DOH-DiF: <LOQ (above 1400 ng L⁻¹) and CP: <LOQ (above 300 ng L⁻¹), and GEM was quantified at 420 ng L⁻¹. Furthermore, 2-DOH-DiF (116,000 ng L⁻¹) was detected at the highest level in the hospital wastewater. There were no zebrafish deaths at any of the concentrations of the compounds used. However, we observed histological changes, including aneurysms and edema in the gills and areas of necrosis of the liver. In summary, we found higher concentrations of CP, GEM and both metabolites (3-NH₂-F and 2-DOH-DiF) were detected for the first time. There is currently no legislation regarding the discharge of anti-cancer drugs in effluents in Brazil. This study is first to focus on effluents from specific treatments from a large cancer hospital located in small city in Brazil.

Diazinon is a common organophosphate pesticide widely used to control parasitic infections in agriculture. Excessive use of diazinon can have adverse effects on the environment and aquatic animal health. In the present study, the toxic effects of diazinon on the histology, antioxidant, innate immune and intestinal microbiota community composition of crucian carp (Carassius auratus gibelio) were investigated. The results showed that diazinon at the tested concentration (300 μg/L) induced gill and liver histopathological damages. Hepatic total superoxide dismutase (T-SOD), catalase (CAT), and glutathione S-transferase (GST) activities significantly decreased (P < 0.05) by 32.47%, 65.33% and 37.34%, respectively. However, the liver tissue malondialdehyde (MDA) content significantly (P < 0.05) increased by 138.83%. The 300 μg/L diazinon significantly (P < 0.05) downregulated the gene expression of TLR4, MyD88, NF-kB p100 and IL-8 but had no significant effect TNF-α (P = 0.8239). In addition, the results demonstrated that diazinon exposure could affect the intestinal microbiota composition and diversity. Taken together, the results of this study indicated that diazinon exposure can cause damage to crucian carp, induce histopathological damage in gill and liver tissues, oxidative stress in the liver, and innate immune disorders and alter intestinal microbiota composition and diversity.

Microplastics (MPs) are widely found in coastal areas and oceans worldwide. The MPs are environmentally concerning due to their bioavailability and potential impacts on a wide range of marine biota, so assessing their impact on the biota has become an urgent research priority. In the present study, we exposed Crassostrea gigas oysters to irregular MPs of two polymer types (polyethylene (PE) and polyethylene terephthalate (PET)) at concentrations of 10 and 1000 μg L⁻¹ for 21 days. Accumulation of MPs, changes in metabolic enzyme activity, and histological damage were evaluated, and metabolomics analysis was conducted. Results demonstrated that PE and PET MPs were detected in the gills and digestive gland following exposure to both tested concentrations, confirming ingestion of MPs by the organisms. Moreover, both PE and PET MPs inhibited lipid metabolism, while energy metabolism enzyme activities were activated in the oysters. Histopathological damage of exposed oysters was also observed in this study. Integrated biomarker response (IBR) results showed that MPs toxicity increased with increasing MPs concentration, and the toxic effects of PET MPs on oysters was greater than PE MPs. In addition, metabolomics analysis suggested that MPs exposure induced alterations in metabolic profiles in oysters, with changes in energy metabolism and inflammatory responses. This study reports new insights into the consequences of MPs exposure in marine bivalves at environmentally relevant concentrations, providing valuable information for ecological risk assessment of MPs in a realistic conditions.

Exposure to crude oil during spill events causes a variety of pathologic effects in birds, including oxidative injury to erythrocytes, which is characterized in some species by the formation of Heinz bodies and subsequent anemia. However, not all species appear to develop Heinz bodies or anemia when exposed to oil, and there are limited controlled experiments that use both light and electron microscopy to evaluate structural changes within erythrocytes following oil exposure. In this study, we orally dosed zebra finches (Taeniopygia guttata) with 3.3 or 10 mL/kg of artificially weathered Deepwater Horizon crude oil or 10 mL/kg of peanut oil (vehicle control) daily for 15 days. We found that birds receiving the highest dosage experienced a significant increase in reticulocyte percentage, mean corpuscular hemoglobin concentration, and liver mass, as well as inflammation of the gastrointestinal tract and lymphocyte proliferation in the spleen. However, we found no evidence of Heinz body formation based on both light and transmission electron microscopy. Although there was a tendency for packed cell volume and hemoglobin to decrease in birds from the high dose group compared to control and low dose groups, the changes were not statistically significant. Our results indicate that additional experimental dosing studies are needed to understand factors (e.g., dose- and species-specific sensitivity) and confounding variables (e.g., dispersants) that contribute to the presence and severity of anemia resulting from oil exposure in birds.

Thiacloprid is a neonicotinoid insecticide widely exploited in agriculture and easily mobilized towards aquatic environments by atmospheric agents. However, little information about its toxicological effects on aquatic invertebrate bioindicators is available. In this study, specimens of the mussel Mytilus galloprovincialis were exposed to thiacloprid at environmental (4.5 μg L⁻¹) and 100 times higher than environmental (450 μg L⁻¹) concentrations for 20 days. Thiacloprid affected haemolymph biochemical parameters, cell viability in the digestive gland, antioxidant biomarkers and lipid peroxidation in the digestive gland and gills at environmentally relevant concentrations (4.5 μg L⁻¹). In addition, thiacloprid exposure caused histological damage to the digestive gland and gills. Interestingly, the pesticide was detected at levels equal to 0.14 ng g⁻¹ in the soft tissues of sentinels exposed for 20 days to 450 μg L⁻¹ thiacloprid in seawaterμ. Due to its harmful potential and cumulative effects after long-term exposure of M. galloprovincialis, thiacloprid may pose a potential risk to nontarget aquatic organisms, as well as to human health. This aspect requires further in-depth investigation.

Developmental exposures to estrogenic chemicals possibly cause structural and functional abnormalities of reproductive organs in vertebrates. Bisphenol AF (BPAF), a bisphenol A (BPA) analogue, has been shown to have higher estrogenic activity than BPA, but little is known about the effects of BPAF on gonadal development, particularly gonadal differentiation. We aimed to determine whether low concentrations of BPAF could disrupt gonadal differentiation and subsequent development using Xenopus laevis, a model species for studying feminizing effects of estrogenic chemicals. X. laevis tadpoles were exposed to BPAF (1, 10, 100 nM) or 17β-estradiol (E2, positive control) from stages 45/46 to 53 and 66 in a semi-static exposure system, with a prolonged treatment with the highest concentration to the eighth week post-metamorphosis (WPM8). Gonadal morphology and histology as well as sexually dimorphic gene expression were examined to evaluate the effects of BPAF. All concentrations of BPAF caused changes in testicular morphology at different developmental stages compared with controls. Specifically, at stage 53, BPAF like E2 resulted in decreases in both the size and the number of gonadal metameres (gonomeres) in testes, looking like ovaries. Some of BPAF-treated testes remained segmented and even became discontinuous and fragmented at subsequent stages. Histological abnormalities were also observed in BPAF-treated testes, such as ovarian cavity at stages 53 and 66 and poorly developed seminiferous tubules on WPM8. At the molecular level, BPAF inhibited expression of male highly expressed genes in testes at stage 53. Correspondingly, BPAF, like E2, inhibited cell proliferation in testes at stage 50. All results show that low concentrations of BPAF inhibited testicular differentiation and subsequent development in X. laevis, along with feminizing effects to some degree. Our finding implies a risk of BPAF to the male reproductive system of vertebrates including humans.