The role of ascorbic acid combined exposure on Imidacloprid-induced oxidative stress and genotoxicity in Nile tilapia

Imidacloprid (Imid), a systemic neonicotinoid insecticide, is broadly used worldwide. It is reported to contaminate aquatic systems. This study was proposed to evaluate oxidative stress and genotoxicity of Imid on Nile tilapia (Oreochromis niloticus) and the protective effect of ascorbic acid (Asc). O. niloticus juveniles (30.4 ± 9.3 g, 11.9 ± 1.3 cm) were divided into six groups (n = 10/replicate). For 21 days, two groups were exposed to sub-lethal concentrations of Imid (8.75 ppm, 1/20 of 72 h-LC50 and 17.5 ppm, 1/10 of 72 h-LC50); other two groups were exposed to Asc (50 ppm) in combination with Imid (8.75 and 17.5 ppm); one group was exposed to Asc (50 ppm) in addition to a group of unexposed fish which served as controls. Oxidative stress was assessed in the liver where the level of enzymatic activities including superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) in addition to mRNA transcripts and, Lipid peroxidation (LPO) were evaluated. Moreover, mitotic index (MI) and comet assay were performed, in addition, the erythrocytic micronucleus (MN), and nuclear abnormalities (NA) were observed to assess genotoxicity in fish. Imid exposure induced significant (p ˂ 0.05) changes in the antioxidant profile of the juveniles' liver by increasing the activities and gene expression of SOD, CAT and GPX as well as elevating the levels of LPO. DNA strand breaks in gill cells, erythrocytes and hepatocytes along with erythrocytic MN and NA were also significantly elevated in Imid-exposed groups. MI showed a significant (p ˂ 0.05) decrease associated with Imid exposure. Asc administration induced a significant amelioration towards the Imid toxicity (8.75 and 17.5 ppm). A significant protective potency against the genotoxic effects of Imid was evidenced in Asc co-treated groups. Collectively, results highlight the importance of Asc as a protective agent against Imid-induced oxidative stress and genotoxicity in O. niloticus juveniles.

An efficient protocol for chromosome isolation from sterlet (A. ruthenus) embryos and larvae

In the present study, the development of an efficient and feasible protocol for chromosome preparation from sterlet (A. ruthenus) embryos and larvae was carried out. In the established protocol, the mean efficiency of chromosome extraction ranged from 70 to 100%. The average number of recorded metaphases per slide was between 9 to 15. In general, the most satisfactory results were obtained for embryos at 6 dpf and larvae at the age of up to 7 dph. In the 24 dpf group, chromosome isolation was possible without immersion in spindle poison, however; in successive developmental stages, the minimal immersion time exceeded 1.5 hours, regardless of chorionation. Immersion for 14 hours did not compromise the efficacy of chromosome isolation. In the current study, successful chromosome isolation was determined mainly by hypotonization conditions. Younger developmental stages generally require the shortest hypotonization times, whereas older larvae require longer hypotonization times. The optimal hypotonization period is 5-15 minutes for embryos at 24 dpf, 40 minutes for embryos at 4dpf, and 50-60 minutes for fish at 6 dpf-7 dph. The only exception was the 24 hpf group where only blastula cells were used. An additional overnight fixation step significantly enhanced chromosome quality and supported chromosome counting especially in the 24 dpf group. The quality and quantity of chromosome slides were also significantly determined by tissue type, and the slides prepared from heads (gill cells) produced the best results.

Imidacloprid Induces Oxidative Stress and Genotoxicity in Nile Tilapia: The Role of Ascorbic Acid Combined Exposure

Imidacloprid (Imid), a systemic neonicotinoid insecticide, is broadly used worldwide. It is reported to contaminate aquatic systems. This study was proposed to evaluate oxidative stress and genotoxicity of Imid on Nile tilapia (Oreochromis niloticus) and the protective effect of ascorbic acid (Asc). O. niloticus juveniles (30.4 ± 9.3 g, 11.9 ± 1.3 cm) were divided into six groups (n=10/replicate). For 21 days, two groups were exposed to sub-lethal concentrations of Imid (8.75 ppm, 1/20 of 72h-LC50 & 17.5 ppm, 1/10 of 72h-LC50); other two groups were exposed to Asc (50 ppm) in combination with Imid (8.75 & 17.5 ppm); one group was exposed to Asc (50 ppm) in addition to a group of unexposed fish which served as controls. Oxidative stress was assessed in the liver where the level of enzymatic activities including superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) in addition to mRNA transcripts and, Lipid peroxidation (LPO) were evaluated. Moreover, mitotic index (MI) and comet assay were performed, in addition to the erythrocytic micronucleus (MN), and nuclear abnormalities (NA) were observed to assess genotoxicity in fish. Imid exposure induced significant (p˂ 0.05) changes in the antioxidant profile of the juveniles' liver by increasing the activities and gene expression of SOD, CAT and GPX as well as elevating the levels of LPO. DNA strand breaks in gill cells, erythrocytes and hepatocytes along with erythrocytic MN and NA were also significantly elevated in Imid-exposed groups. MI showed a significant (p˂ 0.05) decrease associated with Imid exposure. Asc administration induced a significant amelioration towards the Imid toxicity (8.75 & 17.5 ppm). A significant protective potency against the genotoxic effects of Imid was evidenced in Asc co-treated groups. Collectively, results highlight the importance of Asc as a protective agent against Imid-induced oxidative stress and genotoxicity in O. niloticus juveniles.

Glutathione and its dependent enzymes’ modulatory responses to neonicotinoid insecticide sulfoxaflor induced oxidative damage in zebrafish in vivo

The use of neonicotinoid insecticides has progressively increased worldwide when compared with other insecticide groups. Due to this increase, non-target animal species such as fish are exposed to neonicotinoids from different sources, so they can be accumulated at trophic levels and cause various toxic effects by reaching humans. There are limited studies related to the toxic effects of neonicotinoid sulfoximine insecticides including sulfoxaflor on non-target species. The purpose of the present study was to evaluate the effects of sulfoxaflor on GSH-related antioxidants and to determine oxidative stress-producing effect of sulfoxaflor in the gill of zebrafish ( Danio rerio). For this purpose, three sublethal concentrations of sulfoxaflor 0.87 mg/L (2.5% of 96 h LC50), 1.75 mg/L (5% of 96 h LC50), 3.51 mg/L (10% of 96 h LC50) of sulfoxaflor were exposed to zebrafish for 24, 48, and 96 h. GSH related antioxidants were evaluated by analyzing tGSH levels and GPx, GR, GST specific enzyme activities in the gill of zebrafish. The oxidative damage of sulfoxaflor on gill cells was determined by measuring TBARS levels. The results of this study demonstrated that sulfoxaflor activated GSH related antioxidants by increasing tGSH levels, GPx, GR enzyme activities and by diminishing GST enzyme activity in the gill of zebrafish. Sulfoxaflor also caused oxidative damage in the gill of zebrafish by increasing lipid peroxidation. In conclusion, this study indicated that sulfoxaflor led to oxidative stress and activation of GSH related antioxidants in the gill of zebrafish.

GENOTOXIC EFFECT OF CARBARYL ON GILL CELLS OF CHANNA PUNCTATUS

The genotoxicity of carbaryl was evaluated through Chromosomal Aberration Test on gill cells in freshwater fish Channa punctatus. Fishes were acclimatized in laboratory and divided into control and experimental groups. Two sublethal concentrations of carbaryl (0.1ppm and 0.5ppm) were identified and experimental fishes were exposed to these concentrations for a period of 144 hrs. Chromosomal aberrations were increased in carbaryl treated group, both were greater at higher concentration of carbaryl. These finding indicate that carbaryl is able to cause genotoxic effects in Channa punctatus.

Effect-Based Approach to Assess Nanostructured Cellulose Sponge Removal Efficacy of Zinc Ions from Seawater to Prevent Ecological Risks

To encourage the applicability of nano-adsorbent materials for heavy metal ion removal from seawater and limit any potential side effects for marine organisms, an ecotoxicological evaluation based on a biological effect-based approach is presented. ZnCl2 (10 mg L−1) contaminated artificial seawater (ASW) was treated with newly developed eco-friendly cellulose-based nanosponges (CNS) (1.25 g L−1 for 2 h), and the cellular and tissue responses of marine mussel Mytilus galloprovincialis were measured before and after CNS treatment. A control group (ASW only) and a negative control group (CNS in ASW) were also tested. Methods: A significant recovery of Zn-induced damages in circulating immune and gill cells and mantle edges was observed in mussels exposed after CNS treatment. Genetic and chromosomal damages reversed to control levels in mussels’ gill cells (DNA integrity level, nuclear abnormalities and apoptotic cells) and hemocytes (micronuclei), in which a recovery of lysosomal membrane stability (LMS) was also observed. Damage to syphons, loss of cilia by mantle edge epithelial cells and an increase in mucous cells in ZnCl2-exposed mussels were absent in specimens after CNS treatment, in which the mantle histology resembled that of the controls. No effects were observed in mussels exposed to CNS alone. As further proof of CNS’ ability to remove Zn(II) from ASW, a significant reduction of >90% of Zn levels in ASW after CNS treatment was observed (from 6.006 to 0.510 mg L−1). Ecotoxicological evaluation confirmed the ability of CNS to remove Zn from ASW by showing a full recovery of Zn-induced toxicological responses to the levels of mussels exposed to ASW only (controls). An effect-based approach was thus proven to be useful in order to further support the environmentally safe (ecosafety) application of CNS for heavy metal removal from seawater.

Screening of Chilean fish-killing microalgae using a gill cell-based assay

Fish-killing algal species are responsible for important global economic losses to the finfish industry. Chile is the world’s second-largest salmon exporter, and fish-killing algal blooms have widely impacted its production in the last decades. The lack of standardized analytical methods to quantify and characterize the so-called “ichthyotoxins” has hindered our understanding of the underlying ichthyotoxic modes of action. The novel application of a highly sensitive and reproducible fish RTgill-W1 cell line-based assay has allowed significant progress in the field. In this study, the ichthyotoxic potency of the main microalgae species, which has been reported in fish-killing events in the historical Chilean monitoring programs, was assessed. The dinoflagellate Karenia selliformis was the most ichthyotoxic species against the RTgill-W1 (cell viability down to 8%), representing the major threat for the local salmon industry. In comparison, the raphidophyte Heterosigma akashiwo and the dinoflagellate Prorocentrum micans were the least toxic (gill cell viability down to 81 and 89%, respectively). Importantly, ichthyotoxic flagellates were more toxic to fish gill cells upon rupture. These results have important implications for the mitigation and management of algal blooms by the salmon industry.

Ultrastructure of intracytoplasmic Rickettsia-like infection of the gills of the teleost Archosargus probatocephalus (Sparidae) in northeastern Brazil

A histopathological survey was conducted to investigate the presence of microparasites in fish Archosargus probatocephalus in a river near Maceió, Brazil. Light microscope observations of fragments of gill showed the presence of small cysts containing numerous myxospores that were morphologically identified as Henneguya. Transmission electron microscopy observations further revealed several gill cells containing groups of prokaryotic cells within large cytoplasmic vacuoles. Each infected host cell displayed a single vacuole containing a variable number of Rickettsia-like cells (up to 11), some of which presented the dumbbell shape characteristic of binary fission. The Rickettsia-like cells were pleomorphic, without a nucleus and with chromatin dispersed in the cytoplasm. They had a thin electron-dense wall of Gram-negative type. The morphology of these prokaryotic was similar to those of the order Rickettsiales and was described as a Rickettsia-like organism. Histopathological evaluation showed that several vacuole membranes had a lysed appearance. Some had ruptured, thus allowing direct contact between the Rickettsia-like organism and the cytoplasm of the host cell. The rupturing of the branchial epithelium may have contributed towards reduction of the surface area of the gills, but it is not possible to say that this was the cause of the host’s death.