The cerium oxide nanoparticles toxicity induced physiological, histological and biochemical alterations in freshwater mussels, Unio crassus.

INTRODUCTION: Releasing of cerium oxide nanoparticles (nano-CeO2) to the nature has increased due to the widespread use in many fields ranging from cosmetics to the food industry. Therefore, nano-CeO2 has been included in the Organization for Economic Co-operation and Development's (OECD) priority list for engineering nanomaterials. In this study, the effects of nano-CeO2 on the freshwater mussels were investigated to reveal the impact on the freshwater systems on model organism. METHODS: First, the chemical and structural properties of nano-CeO2 were characterized in details. Second, the freshwater mussels were exposed to environmentally relevant concentrations of nano-CeO2 as 10 mg, 25 mg and 50 mg/L during 48-h and 7-d. Third, after the exposure periods, hemolymph and tissue samples were taken to analyse the Total Hemocyte Counts (THCs) histology and oxidative stress parameters (total antioxidant status, glutathione, glutathione-S-transferase, and advanced oxidative protein products). RESULTS: Significant decrease of the THCs was observed in the nano-CeO2 exposed mussels compared to the control group (P < 0.05). The histological results showed a positive association between nano-CeO2 exposure concentration in the water and level of tissue damage and histopathological alterations were detected in the gill and the digestive gland tissues. Oxidative stress parameters were slightly affected after exposure to nano-CeO2 (P > 0.05). In conclusion, this study showed that acute exposure of freshwater mussels to nano-CeO2 did not pose significant biological risk. However, it has been proven that mussels are able to accumulate nano-CeO2 significantly in their bodies. CONCLUSION: This suggests that nano-CeO2 may be a potential risk to other organisms in the ecosystem through trophic transfer in the food-web based on their habitat and niche in the ecosystem.

How do biocidals affect the non-target marine organisms: the short-term effects of antifouling agent sodium pyrithione on Mediterranean mussels (Mytilus galloprovincialis, Lamark 1819).

The metallic pyrithiones are used as antifouling paints for marine vehicles against fouling organisms. However, they are dissolved in marine water and have negative impacts on marine non-target organisms. This study evaluated the adverse effects of sodium pyrithione (NaPT) on Mytilus galloprovincialis using total hemocyte counts (THCs), oxidative stress and antioxidant parameters, and histopathological observations. Mussels were exposed to 0.1 and 1 µg/L NaPT for 96 h. The THC values of the NaPT-exposed mussels significantly decreased (p < 0.05). Lipid peroxidation and advanced oxidative protein products of digestive gland and gill tissues were decreased but only the digestive gland tissues of 0.1 µg/L NaPT values were significantly decreased compared to control groups (p < 0.05). Histological alterations were observed in the gill and the digestive gland tissues revealing malformations and hyperplasia of gill lamella; degenerations and loss of tubules of digestive gland after exposure to NaPT for 96 h. As a result, biocidal sodium pyrithione has adverse effects on the mussels even in short-term exposures and low concentrations.

Pyrethroid-induced oxidative stress and biochemical changes in the primary mussel cell cultures.

Pyrethroids are among the most widely used insecticides. Permethrin and tetramethrin, which are synthetic pyrethroids, are generally used to control insects in agricultural areas and household applications. Due to broad use areas, they contaminate aquatic ecosystems and cause adverse effects to the non-target aquatic organisms. Even though permethrin and tetramethrin are known to alter the oxidative stress parameters of in vivo aquatic animal model organisms, there are limited studies in vitro. This study aims to determine the adverse effects of permethrin and tetramethrin in the in vitro models of freshwater mussels exposed to 1 mg/L, 10 µg/L, 100 ng/L and 1 ng/L concentrations of chemicals for 24 h. For this purpose, reduced glutathione activities were evaluated as biomarkers of the primary gill and digestive gland cell cultures. In both cell cultures, reduced glutathione values increased in the exposed groups, compared to the control group. Even though the results showed that reduced glutathione activities had not significantly changed concentration-dependently (p > 0.05), significant differences were observed in the reduced glutathione activities of both cell cultures (p < 0.05). This study showed that permethrin and tetramethrin had highly toxic effects in the in vitro models of mussels even at low concentrations.

Are BPA-free plastics safe for aquatic life? - Fluorene-9-bisphenol induced thyroid-disrupting effects and histopathological alterations in adult zebrafish (Danio rerio).

Fluorene-9-bisphenol (BPFL) is used as an alternative compound for bisphenol A, an endocrine disruptor compound which is present in various materials including plastic bottles and packaging. Although it is used extensively in products that are labelled BPA-free, its effect on wildlife and humans have not been fully studied. Therefore, this study aimed to investigate the effects of BPFL in adult zebrafish. In the preliminary experiments of the study, the median lethal concentration value (LC50) of BPFL was 0.25 mg/L (95 % confidence interval 0.15-0.41) for 96 h. Following exposure to three different sublethal concentrations of BPFL after 96 h and 15 days, T4 hormone levels, expression levels of genes involved in thyroid metabolism and histopathological alterations were assessed. T4 hormone levels were found to be significantly higher in females at the lowest BPFL concentration following 96 h exposure (P < 0.05). Expression levels of trh, tshba and trhrb genes were upregulated following 96 h exposure at 0.025 mg/L concentration and crh was upregulated following 15 days exposure at 0.025 mg/L concentration in female zebrafish (P < 0.05). The most prominent histopathological findings in zebrafish exposed to 0.025 and 0.125 mg/L of BPFL were observed in the gill, liver, kidney and testis tissues. The gill tissues showed some hyperemia, lamellar fusion, hyperplasia, epithelial lifting, and telangiectasis, while passive hyperemia, hydropic degeneration, and necrosis were observed in the liver tissues. The BPFL is highly toxic to zebrafish even in sublethal concentrations according to the molecular and histopathological responses.

The effects of invasive epibiont Dreissena polymorpha on the narrow-clawed crayfish (Astacus leptodactylus, Astacidae) in Egirdir Lake, Turkey: a case study.

In aquatic ecosystems, organisms are in a close relationship with each other. While this relationship is sometimes beneficial for both species, it can sometimes be harmful to one species. This study aimed to investigate the effects of colonized zebra mussels Dreissena polymorpha on the growth parameters and total hemocyte counts of the narrow-clawed crayfish Astacus leptodactylus from Egirdir Lake (Turkey). Zebra mussels were found on various body parts of crayfish and the most intense (44.7%) colonization was in the carapace region. There were significant differences between various biometric data and ratios of the narrow-clawed crayfish with epibiont D. polymorpha and without epibiont D. polymorpha (the control group) in terms of total length, abdomen width, arm length and chelae length, carapace width/carapace length, carapace length/abdomen length, abdomen width/carapace width, and chelae width/chelae length (p < 0.05). When the total hemocyte counts were examined, lower values ​were obtained in the narrow-clawed crayfish with zebra mussels compared to the control group (p < 0.05). As a result, epibiont organisms have negative effects on the health and growth status of the host organisms considering total hemocyte counts and biometric data.

Physiological and histopathological alterations in Capoeta baliki and Squalius pursakensis after caused by some environmental pollutants.

The present study was aimed to apply physiological and histological biomarkers of pelagic and benthic fish species as biomonitoring of environmental pollutants. Capoeta baliki and Squalius pursakensis were caught from 5 stations in Kirmir Stream (Turkey). After measuring the total length and weight for condition factor analysis, liver, gill, brain, and kidney tissues of fish were taken for histopathological analysis. Dorsal muscles of fish tissues were analyzed for polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), organochlorine pesticides (OCPs), and polybrominated diphenyl ethers (PBDEs). The mean condition factor of C. baliki and S. pursakensis were 1.14 and 0.97, respectively. It was indicated that the first one was in good condition than the latter one. According to histopathological observations, the alterations in the tissues may be associated with the environmental pollutants' exposure. The total concentration of PAHs of dorsal muscle of C. baliki and S. pursakensis found ranged from 0.915 to 108.421 µg/kg and 0.601 to 675.248 µg/kg, respectively. The highest total concentrations of OCPs were found as 44.16 µg/kg for C. baliki and as 562.12 µg/kg for S. pursakensis. The PCB 28 (10.83 µg/kg), PCB 52 (10.92 µg/kg), PCB 138 (226.24 µg/kg), and the total concentrations of PBDEs were found as 151.74 µg/kg in only S. pursakensis. The result of the concentrations of the investigated pollutants was generally found higher in S. pursakensis which is a higher trophic level than C. baliki. Histopathological alterations were detected as hyperemia in liver, gill, and brain; as hydrophobic degeneration in liver; hyperplasia in gill; and tubular degeneration in kidney tissues of both species. The obtained result of the study shows that the environmental pollutants may affect the physiological and histological status of fish.

How does sublethal permethrin effect non-target aquatic organisms?

Permethrin is belonged to pyrethroids that are one of the substances developed as an alternative to pesticides. Permethrin, which is used especially in agriculture, can bioaccumulate in the water and sediment when mixed into aquatic ecosystems. For this reason, it is necessary to investigate the effect of this substance on aquatic organisms other than the target organism. The aim of this study was the determination of acute and sublethal effects as antioxidant enzyme levels on different organs and hemolymph biochemistry of the non-target aquatic organism, narrow-clawed crayfish (Astacus leptodactylus), after exposure to permethrin, one of the synthetic pyrethroid pesticides, contaminating aquatic ecosystems due to its increase usage. The invertebrate model organism, the narrow-clawed crayfish, was selected for its bioindicator role in food webs as planktivorous grazers epibenthic scavengers and good alternative models in ecotoxicology studies with the importance in conservation of freshwater ecosystems. The 96-h LC50 value of permethrin to experimental species was estimated as 0.903 µg/L (95% CI = 0.5042-2.2734 µg/L) with probit analysis method. The sublethal concentration of the permethrin was determined by 1/10 of 96-h LC50 values as 0.09 µg/L. There were two control (negative and acetone) groups in the experiment. The sampling of hemolymph and the tissues (gills, hepatopancreas, and muscle) were done 48 h and 96 h after exposure of the permethrin. The total hemocyte counts significantly increased in the 96-h exposed group of permethrin (p<0.05). Among the hemolymph biochemical parameters, the hemolymph potassium and chloride values increased statistically (p<0.05). Malondialdehyde levels (MDA) of gills and muscle were significantly increased, whereas the MDA level of the hepatopancreas was significantly decreased at the end of the experiment (p<0.05). Hyperplasia in the lamella was recorded in gills, while the degenerations of the hepatopancreas tissues were observed. According to obtained results, permethrin was extremely toxic as acutely to narrow-clawed crayfish and also effected at sublethal concentrations.

In vitro effects of erythromycin and florfenicol on primary cell lines of Unio crassus and Cyprinus carpio.

The ubiquitous use of antibiotics leads exposure of these chemicals on non-target aquatic species, while the toxicity assays for these chemicals are time/labor consuming and expensive. Alternative approaches using primary cell cultures which retain the tissue functionality at its highest form have received global attention compared to cell lines. In the current study, the cytotoxic effects of two commonly used antibiotics from amphenicol (florfenicol) and macrolide (erythromycin) groups were evaluated on primary cell cultures of Unio crassus (mantle, digestive gland, gill, and gonad) and Cyprinus carpio (gill and liver) using MTT and Neutral Red assays. The highest cytotoxic effects were found on the mussel digestive gland and carp liver cells for florfenicol and erythromycin, while the lowest cytotoxic effects were found in mussel mantle cells for both drugs in the MTT test. In the NR test, the highest cytotoxic effects of erythromycin and florfenicol were found in the mussel gill, mantle, gonad, and carp gill cells; the lowest cytotoxic effect of erythromycin was found in the mussel digestive gland, while the lowest effect of florfenicol was found in the carp liver cells. The cytotoxicity of florfenicol was quite low for the carp liver, while the cytotoxicity of erythromycin was quite low in the mussel digestive tract. Thus, it was concluded that cells made from mussel tissues could be used in ecotoxicity tests, and sensitivity may vary according to the tissue.

Companion animals get close to the toxic aspects of antropogenic world: cytotoxicity of phthalates and bisphenol A on dog testicular primary cells.

Phthalates, which are among the most abundant plasticizers, have detrimental effects on the reproductive system. Similar to human, dogs are prominently exposed to phthalates in daily routines at low concentrations; while toys, training devices and commercial dog foods are considered as the primary sources of exposure. This study aimed to reveal and compare the cytotoxic effects of selected phthalates (Benzyl butyl phthalate (BBP), Diethyl phthalate (DEP), Bis (2-ethylhexyl) phthalate (DEHP), Di-'isobutyl' phthalate (DIBP), Di-'isodecyl' phthalate (-DIDP) Di-'isononyl' phthalate (DINP), Dimethyl phthalate (DMP), Di-n-octyl phthalate (DNOP)), and Bisphenol A (BPA) following 24 h exposure on primary testicular parenchymal cells of dog in vitro at concentrations between 0.001 and 2.5 nM. According to cytotoxicity results, DEHP was found to be the most toxic phthalate with IC50 at 22.53 µM; while DMP was the least (169.17 nM). IC50 of BPA was 161.81 nM, less than the average (61.95 nM) of phthalates. In addition, dog primary testicular cells were found more susceptible to the high molecular weight phthalates (DNOP, DEHP, DINP, DIDP) than low molecular weight phthalates (DMP, DEP, DIBP, BBP). Further studies should focus on morphological, physiological and molecular differences to comprehend the mechanisms involved as well as decreasing the risk for impaired spermatogenesis caused by environmental toxicants in companion animal medicine.

Ocular surface findings in patients with rheumatoid arthritis under methotrexate or biological agent therapy.

PURPOSE: To evaluate the ocular findings in patients with rheumatoid arthritis (RA) treated with disease-modifying antirheumatic drugs (DMARDs) such as methotrexate (MTX) or MTX with biological agents. METHODS: One hundred and twelve eyes of 56 patients with RA and treated with MTX or MTX with biological agents were included in the study. Patients were divided into two groups using DMARDs only (group 1) and patients using DMARDs and biologic agents together (group 2). In both groups; Schirmer's II test, tear film break-up time (tBUT), central corneal thickness (CCT), corneal volume (CV), intraocular pressure (IOP) measurement, and anterior segment and fundus examinations of the eye with slit lamp were carried out. Ocular surface disease index (OSDI) score questionnaire were performed. RESULTS: Thirty-eight patients with a mean age of 53.00 ± 8.19 years were in group 1 and 18 patients with a mean age of 51.00 ± 9.54 years were in group 2. The mean duration of RA was 6.89 ± 7.96 years in group 1 and 5.70 ± 9.00 years in group 2. There was a statistically significant difference between two groups with tBUT, CCT, CV, IOP (p < 0.05) and there was no significant difference with age, sex, disease duration, disease activity, and Schirmer's II test (p > 0.05). The disease duration showed a significant moderate negative correlation with CCT and CV in group 2 (p < 0.05). CONCLUSIONS: Although tBUT values were significantly higher in the combination treatment group, CCT and CV values were significantly lower. Due to the decrease in corneal thickness, IOP was determined to be significantly lower.