Erythrocytic Nuclear Abnormalities Research Articles

Exposure of adult zebrafish (Danio rerio) to SARS-CoV-2 at predicted environmentally relevant concentrations: Outspreading warns about ecotoxicological risks to freshwater fish

While the multifaceted social, economic, and public health impacts associated with the COVID-19 pandemic are known, little is known about its effects on non-target aquatic ecosystems and organisms. Thus, we aimed to evaluate the potential ecotoxicity of SARS-CoV-2 lysate protein (SARS.CoV2/SP02.2020.HIAE.Br) in adult zebrafish (Danio rerio) at predicted environmentally relevant concentrations (0.742 and 2.226 pg/L), by 30 days. Although our data did not show locomotor alterations or anxiety-like or/and anxiolytic-like behavior, we noticed that exposure to SARS-CoV-2 negatively affected habituation memory and social aggregation of animals in response to a potential aquatic predator (Geophagus brasiliensis). An increased frequency of erythrocyte nuclear abnormalities was also observed in animals exposed to SARS-CoV-2. Furthermore, our data suggest that such changes were associated with a redox imbalance [↑ROS (reactive oxygen species), ↑H2O2 (hydrogen peroxide), ↓SOD (superoxide dismutase), and ↓CAT (catalase)], cholinesterasic effect [↑AChE (acetylcholinesterase) activity], as well as the induction of an inflammatory immune response [↑NO (nitric oxide), ↑IFN-γ (interferon-gamma), and ↓IL-10 (interleukin-10)]. For some biomarkers, we noticed that the response of the animals to the treatments was not concentration-dependent. However, principal component analysis (PCA) and the “Integrated Biomarker Response” index (IBRv2) indicated a more prominent ecotoxicity of SARS-CoV-2 at 2.226 pg/L. Therefore, our study advances knowledge about the ecotoxicological potential of SARS-CoV-2 and reinforces the presumption that the COVID-19 pandemic has negative implications beyond its economic, social, and public health impacts.

Description and quantification of micronucleus and nuclear abnormalities in erythrocytes of the sentinel green turtle (Chelonia mydas) with fluorescence microscopy.

Contamination in marine ecosystems is of the most critical threats to marine turtles. The identification of useful biomarkers to detect and monitor the physiological and clinical effects of pollutants on these populations will allow early detection of alterations (e.g., mutagenic damages) that could risk their viability or favor the development of diseases, thus threatening the biodiversity of these ecosystems and human population. This study is aimed at describing and quantifying nuclear anomalies in peripheral blood erythrocytes of green turtles (Chelonia mydas) from three distinct foraging areas in Mexico (Akumal, Xcalak, and Punta Herrero). We developed a novel morphological index that could be used as a biomarker to identify abnormal nuclei in peripheral blood erythrocytes. Here we describe for the first time in C. mydas, with a species-specific staining protocol, distinct nuclear abnormalities such as blebbed, lobed, notched, eight shape nuclei, and binucleated cells. These nuclear abnormalities were present in >90 % of the subjects (n=30). Moreover, 50 % of the organisms presented erythrocytes with micronuclei. The number of nuclear abnormalities did not correlate with size of the green turtles or differ between sites, or health status. We found a higher frequency of green turtles with nuclear abnormalities in the southern region (Punta Herrero and Xcalak) with the highest frequency of micronucleus and buds. The former could be associated to the constant exposure to chemical pollutants of oceanographic origin in the southern coast of Quintana Roo. Furthermore, the increasing anthropogenic pollution in Akumal could also explain the highest variability in the number of nuclear abnormalities presented in resident individuals. We propose that a long-term monitoring programs of green turtle populations in the Mexican Caribbean that include a micronucleus test could be a useful to determine possible mutagenic damage in these animals.

Oxidative Stress in a Wild Population of the Freshwater Fish Hyphessobrycon Luetkenii Chronically Exposed to a Copper Mining Area: New Insights into Copper Toxicology.

In this study, we examine markers of oxidative stress in the tetra Hyphessobrycon luetkenii collected from two locations in the copper contaminated João Dias creek (southern Brazil). Also, specimens were translocated from a clean reference section of the creek to a polluted stretch and vice-versa. Fish were held at in submerged cages for 96h and then sacrificed. Nuclear abnormalities in erythrocytes and total antioxidant capacity, lipid peroxidation and protein carbonylation in gills, brain, liver and muscle displayed similar trends in both groups. Lipid peroxidation increased in all tissues of individuals translocated to the polluted site but only in liver and muscle of those translocated to the reference site. Increased protein carbonylation was also observed in gills of individuals translocated to the reference location. These results suggest similar oxidative stress among fish from the reference and polluted locations and that long-term metals exposure may require adaptations toward oxidative stress responses.

Potential ecotoxicity of substrate-enriched zinc oxide nanoparticles to Physalaemus cuvieri tadpoles

Although the ecotoxicological effects of ZnO nanoparticles (ZnO NPs) have already been reported in different taxa, little is known about their impacts on amphibians. Thus, we aimed to evaluate the potential effects of exposure of Physalaemus cuvieri tadpoles to substrates enriched with ZnO NPs (and with its ionic counterpart, Zn+2, ZnCl2 – both at 100 mg/kg) previously used in the cultivation of Panicum maximum (Guinea grass). We showed that although exposure for 21 days did not impact the survival, growth, and development of tadpoles, we noted an increase in the frequency of erythrocyte nuclear abnormalities in the “ZnCl2” and “ZnONP” groups, which was associated with suppression of antioxidant activity in the animals (inferred by SOD and CAT activity and DPPH free radical scavenging capacity). In the tadpoles of the “ZnONP” group, we also noticed a reduction in creatinine and bilirubin levels, alpha-amylase activity, and an increase in alkaline phosphatase activity. But the treatments did not alter the activity of the enzymes lactate dehydrogenase and gamma-glutamyl-transferase and total protein and carbohydrate levels. On the other hand, we report a cholinesterase and hypotriglyceridemic effect in the “ZnCl2” and “ZnONP” groups. Zn bioaccumulation in animals, from ZnO NPs, from Zn+2 released from them, or both, has been associated with causing these changes. Finally, principal component analysis (PCA) and the values of the “Integrated Biomarker Response” index revealed that the exposure of animals to substrates enriched with ZnO NPs caused more pronounced effects than those attributed to its ionic counterpart. Therefore, our study reinforces the need to consider the environmental risks of using these nanomaterials for agricultural purposes for amphibians.

Brodifacoum Levels and Biomarkers in Coastal Fish Species following a Rodent Eradication in an Italian Marine Protected Area: Preliminary Results.

Brodifacoum is the most common rodenticide used for the eradication of invasive rodents from islands. It blocks the vitamin K cycle, resulting in hemorrhages in target mammals. Non-target species may be incidentally exposed to brodifacoum, including marine species. A case study conducted on the Italian Marine Protected Area of Tavolara Island was reported after a rodent eradication using the aerial broadcast of a brodifacoum pellet. Brodifacoum presence and effects on non-target marine organisms were investigated. Different fish species were sampled, and a set of analyses was conducted to determine vitamin K and vitamin K epoxide reductase concentrations, prothrombin time, and erythrocytic nuclear abnormalities (ENA) assay. In all the examined organisms, brodifacoum was not detected. The results obtained showed differences in vitamin K and vitamin K epoxide concentrations among the samples studied, with a positive correlation for three species between vitamin K, vitamin K epoxide, and fish weight. The prothrombin time assay showed a good blood clotting capacity in the fish. Higher abnormality values were recorded for four species. The results of this study suggest that it is possible to hypothesize that the sampled fish were not likely to have been exposed to brodifacoum and that consequently there are no negative issues concerning human consumption.

Ethoxysulfuron causes nuclear abnormalities in erythrocytes, DNA damage in some visceral organs, and oxidative stress in male Japanese quail

Ethoxysulfuron causes nuclear abnormalities in erythrocytes, DNA damage in some visceral organs, and oxidative stress in male Japanese quail

Contraceptive-Pill-Sourced Synthetic Estrogen and Progestogen in Water Causes Decrease in GSI and HSI and Alters Blood Glucose Levels in Climbing Perch (Anabas testudineus)

The present study was conducted to understand the changes in gonads and hematological parameters in climbing perch (Anabas testudineus) exposed to synthetic estrogen and progestogen [mixture of ethinylestradiol (EE2) and desogestrel (DES)]. Climbing perch were exposed to four different concentrations of EE2/DES mixtures, viz. 0 ng of EE2 and DES/L (T0), 3 ng EE2 and 15 ng DES/L (T3), 30 ng EE2 and 150 ng DES/L (T30), and 300 ng EE2 and 1500 ng DES/L (T300) for 60 days. On days 45 and 60, samples were taken to assess changes in somatic indexes, gonad histology, and hematological parameters. The gonadosomatic index (GSI) increased in both females and males with increasing concentrations of estrogen mixtures except for T30 females, which was the lowest among all\four treatments. The hepatosomatic index (HSI) was observed to be increased in males as estrogen content increased. However, compared to fish at T0, HSI in female individuals did not vary in T30 fish, where the value was the highest among all the treatments. On day 45, histological observations showed no feminization or intersexuality but several germ-cell deformities in the ovary (adhesion, degenerated oocyte wall, degenerated granulose layer, increased interfollicular space, atretic follicle, and cytoplasmic clumping) and testes (increased interstitial area, focal loss of spermatocyte, dilation of the lumen, breakage of tubular epithelium, and elongated seminiferous tubule) were observed in fish exposed to EE2 and DES. Fish reared at T30 had lower RBC count, hemoglobin (Hb), glucose, and hematocrit levels. On day 60, fish reared at T30 had the highest Hb content compared to fish raised in other treatment conditions. WBC was progressively higher with increasing EE2/DES concentrations. Significant erythrocyte cytoplasmic abnormalities and erythrocyte nuclear abnormalities were observed in fish exposed to higher EE2/DES concentrations. The present study provides insights into the adverse impacts of synthetic estrogens sourced from human contraceptive pills on fish physiology.

Acute and sub-lethal toxicity in African mud catfish (Clarias gariepinus, Burchell, 1822) exposed to some pesticides

Pesticides from agricultural run-off pose a severe threat to non-target organisms, such as fishes. This study was carried out to evaluate acute toxicity and histological and genotoxic effects (erythrocytic nuclear abnormalities) of lethal and sub-lethal concentrations of three commonly used pesticides: Atrazine, Butachlor and Glyphosate on the African mud catfish (Clarias gariepinus). Fishes were exposed to the pesticides for 96h periods to determine their LC50 and the sub-lethal effect at various concentrations (1/10th, 1/100th , 1/1000th 96h LC50) over 28 days. The 96h LC50 values were 7.63mg/l, 0.7mg/l and 15.97mg/l for atrazine, butachlor and glyphosate, respectively. Histological sections of the liver of C. gariepinus exposed to the three pesticides showed mild vascular congestion but no necrosis in the tissue. There was a significant (p<0.5) dose-dependent increase in micronuclei and nuclear abnormalities in the erythrocytes of exposed C. gariepinus compared to the control by 28 days. The study confirmed that C. gariepinus are at risk of adverse effects from exposure to pesticides. Discharge of agricultural run-off around water bodies should be prevented or prohibited to avoid adverse effects on aquatic life.

Effects on liver somatic index, erythrocyte nuclear abnormalities and biliary metabolites of PAH in Oreochromis niloticus exposed to water-borne crude oil

Effects on liver somatic index, erythrocyte nuclear abnormalities and biliary metabolites of PAH in Oreochromis niloticus exposed to water-borne crude oil

Mercury bioaccumulation, genotoxic and biochemical biomarkers reveal the health status of yellow-spotted Amazon River turtles (Podocnemis unifilis) in an environmental protection area in the Amazon

ABSTRACT Chelonians are considered good bioindicators of environmental quality. The assessment of the health status of chelonian populations in the Amazon is important because they are traditionally consumed in large numbers in riverine communities and sustainable use reserves. The present study aimed to evaluate the health of Podocnemis unifilis (Testudines, Podocnemididae) in an environmental protection area in the Amazon region in Brazil. We analyzed the biomarkers lipoperoxidation, carbonylation of proteins, occurrence of micronuclei and erythrocytic nuclear abnormalities, quantified metallothioneins, and evaluated mercury bioaccumulation. We generated pioneering data regarding biomarkers in wild Amazonian freshwater turtles. All biomarker responses did not vary significantly between the sexes. The occurrence of oxidative and genotoxic damage, as well as concentrations of metallothioneins was low compared to other studies. In addition, the bioaccumulation of mercury in the muscle of the animals was below the limit recommended for human consumption by the World Health Organization. Our results provide baseline data on mercury bioaccumulation and biomarker responses that can be useful for future comparisons with other freshwater turtles. The data also provide evidence that the sustainable exploitation of these turtles in the study area (Piagaçu-Purus Sustainable Development Reserve) does not pose a risk to local riverine communities, as the detected mercury concentrations are safe for human consumption. In this sense, our results highlight the importance of protected areas for the conservation of healthy turtle populations, at the same time ensuring the health of the human populations that use them as a food resource.

Hepatic and blood alterations in Lithobates catesbeianus tadpoles exposed to sulfamethoxazole and oxytetracycline

In this study the effects of environmentally realistic concentrations of the antibiotics sulfamethoxazole (SMX) and oxytetracyclyne (OTC) on Lithobates catesbeianus tadpoles were evaluated, through the analyzes of the frequencies of micronucleus and nuclear abnormalities in erythrocytes, alterations in leucocytes, liver histopathology, and changes in hepatic esterase activities and oxidative stress biomarkers. The animals were exposed for 16 days at concentrations of 0 (control), 20, 90 and 460 ng L−1. No significant difference was found in the frequencies of micronucleus and nuclear abnormalities. The two highest concentrations of SMX and all concentrations of OTC caused a significant increase in the number of lymphocytes. A significant decrease in the number of neutrophils compared to the control group was observed for all concentrations tested of both antibiotics. Also, decrease in the activity of glutathione S-transferase and high histopathological severity scores, indicating liver damage, were found in tadpoles exposed to the two highest concentrations of SMX and all concentrations of OTC. The main changes in the liver histopathology were the presence of inflammatory infiltrate, melanomacrophages, vascular congestion, blood cells and eosinophils. Esterase activities were unchanged. Indeed, the two highest concentrations of OTC caused a reduction in the activities of superoxide dismutase and glucose 6-phosphate dehydrogenase, while the highest concentration inhibited the activity of glutathione peroxidase and increased protein carbonyl levels. These results evidences that environmentally realistic concentrations of SMX and OTC in aquatic environments are capable to significantly disrupt tadpoles’ physiology, possibly affecting negatively their survival rate in natural environments.

Toxicity and genotoxicity of imidacloprid in the tadpoles of Leptodactylus luctator and Physalaemus cuvieri (Anura: Leptodactylidae)

Imidacloprid is a neonicotinoid insecticide used to control agricultural pests around the world. This pesticide can have adverse effects on non-target organisms, especially in aquatic environments. The present study evaluated the toxicity of an imidacloprid-based insecticide in amphibians, using Leptodactylus luctator and Physalaemus cuvieri tadpoles as study models. Spawning of both species were collected within less than 24 h of oviposition from a non-agricultural land at Erechim, Rio Grande do Sul state, Brazil. Survival, swimming activity, body size, morphological malformations, and genotoxic parameters were analyzed at laboratory conditions. A short-term assay was conducted over 168 h (7 days) with five different concentrations of imidacloprid (3–300 µg L−1) being tested. The insecticide did not affect survival, although the tadpoles of both species presented reduced body size, malformed oral and intestine structures, and micronuclei and other erythrocyte nuclear abnormalities following exposure to this imidacloprid-based compound. Exposure also affected swimming activity in L. luctator, which reflected the greater sensitivity of L. luctator to imidacloprid in comparison with P. cuvieri. The swimming activity, body size, and malformations observed in L. luctator and the morphological malformations found in P. cuvieri indicated that even the lowest tested concentration of the insecticide were harmful to amphibians. At concentrations of over 3 μg L−1, P. cuvieri presents a smaller body size, and both species are affected by genotoxic cell damage. This demonstrates that imidacloprid is potentially toxic for the two study species at environmentally relevant concentrations.

Fish biomarker responses reflect landscape anthropic disturbance in savanna streams.

Disturbance in the landscape surrounding streams can interfere with water quality and cause harm to aquatic organisms. In this study, we evaluate the influence of land use on the genetic and biochemical biomarkers of fish in streams of Brazilian savanna (Cerrado). We also evaluated whether biomarker responses are seasonally consistent. For this purpose, individuals of the Neotropical tetra fish Astyanax lacustris were exposed in cages for 96h, in 13 streams draining agroecosystems with different degrees of disturbance during the dry and wet seasons. After exposure, blood, liver, and gills were collected for multibiomarker analyses (micronuclei, erythrocytic nuclear abnormalities, lipid peroxidation, antioxidant enzymes, and biotransformation enzyme). The results showed that the gradient of anthropic disturbance was positively associated with genotoxic damage (erythrocytic nuclear abnormalities) and negatively associated with antioxidant and biotransformation enzymes of the liver in both seasons. No association of the gradient of anthropic disturbance with the frequency of micronuclei and for most gill enzymes was found for both seasons. Landscape disturbance was also negatively associated with water quality in the wet season. These results indicate that changes in land use interfere with the genetic and biochemical processes of organisms. Thus, the multibiomarker approach may represent an effective strategy for assessing and monitoring terrestrial landscape disturbance.

Nanoplastics are bioaccumulated in fish liver and muscle and cause DNA damage after a chronic exposure

The extent of the widespread, planetary contamination by plastic waste is difficult to fully capture. Nanoplastics (NPs) are currently in the center of research concerning plastic litter, both for the analytical challenges they pose and for their potential to provoke hazardous effects in organisms. However, there are still many unanswered questions in this multidisciplinary field, with a crucial missing piece being the quantification of NPs in fish tissues after in vivo exposures. Another relevant question that is still greatly unexplored is how a chronic exposure to NPs will affect fish health. This study aims to provide answers to both of these relevant knowledge gaps. To this end, goldfish (Carassius auratus) were exposed to 44 nm polystyrene (PS)-NPs via water for 30 days. Following the exposure, gastrointestinal tract, liver and muscle were sampled for PS-NPs analysis by means of size exclusion chromatography coupled to high resolution mass spectrometry. PS-NPs were detected in all liver and muscle samples of exposed fish, with higher concentrations in liver than in muscle, whereas no PS-NPs were detected in the gastrointestinal tract. Nevertheless, exposure to PS-NPs did not induce changes in hematology parameters nor in cortisol and glucose levels in plasma. On the other hand, even a relatively low concentration of PS-NPs was able to cause DNA damage, measured by an increase in erythrocyte nuclear abnormalities, suggesting that PS-NPs can reach the cell nucleus and cause genotoxicity. These results show for the first time that PS-NPs find their way to fish muscle after chronic exposure, where they bioaccumulate, but do not alter fish survival nor hematological or physiological stress indicators. The accumulation of PS-NPs in fish muscle can represent a threat to human health as a possible route of exposure to small-sized plastics. The present results in a model fish species open windows for future studies in edible fish species.

In vivo assessment of genotoxic effects in Cyprinus carpio L., 1758 (Teleostei: Cyprinidae) exposed to selected metal(oid)s.

Heavy metals and metalloids originating from industrial, agricultural, and urban wastes and increasing in aquatic ecosystems cause genotoxic damage to fish species. This study aimed to determine the potential genotoxic effects of mixtures of aluminum, arsenic, and manganese in Cyprinus carpio. The effects of the mixtures on erythrocyte cells of C. carpio were examined using the comet assay, micronucleus test, and erythrocytic nuclear abnormalities in two groups after exposure to three doses of the mixtures (Group A; Dose 1: 0.3 + 0.1 + 0.02mg/L, Dose 2: 0.6 + 0.2 + 0.04mg/L, Dose 3: 0.9 + 0.3 + 0.06mg/L and Group B; Dose 1: 1 + 3 +0.1mg/L, Dose 2: 2 + 6 + 0.2mg/L, Dose 3: 3 + 9 + 0.3mg/L). Experimental groups were formed according to the permissible limits specified in the Turkish Surface Water Quality Regulation (TSWQR). The results of comet assay parameters such as tail DNA %, tail moment, and olive tail moment confirmed the genotoxic effect of metal(oid)s mixtures on erythrocyte cells compared with control groups and showed that DNA damage increased with increasing the concentrations. The micronucleus and other nuclear abnormalities such as blebbed nuclei, notched nuclei, eightshaped nuclei, lobed nuclei, and binucleated cells were detected in the erythrocyte cells exposed to the mixtures. Consequently, it was found that the frequency of micronucleus and erythrocytic nuclear abnormalities significantly increased in the erythrocyte cells exposed to metal(oid) concentrations compared to control groups. These results show the existence of potential genotoxicity in C. carpio even at the minimum values specified in the TSWQR after exposure to the mixtures.

Fish feed can show genotoxic damage.

The present study was carried out to evaluate the genotoxic potential of nutritional quality of feed, using erythrocytic nuclear abnormalities assay in Nile tilapia and its correlation with available nutrients and common fish growth biomarkers. For this, ten feeds commercialized in Brazil were assessed on digestibility and performance assays with triplicate groups. Venipuncture of the caudal vein for abnormalities analyzed was performed 102days after the fish were fed with the commercial feed twice a day, to apparent satiation. Nuclear abnormalities were analyzed in blood smears. Principal component analysis and correlation matrix were used to carry out an exploratory analysis of correlation between frequency of abnormalities and performance parameters or available nutrients. The frequency of abnormalities in erythrocytes of Nile tilapia, fish performance, and digestibility were feed-dependent. It was observed correlations between the frequency of most abnormalities and performance parameters or content of nutrients. The frequency of kidney-shaped (6.23 ± 4.14), bud nuclei (2.99 ± 1.95), bridge nuclei (0.53 ± 0.50), and binuclei (0.43 ± 0.59) was the highest in fish that also presented the worst performance among all treatments and correlated with digestible methionine. Micronucleus frequency (0.33 ± 0.49) was higher in fish from the same group that presented depressed feed intake and lower available zinc. Only one group presented vacuolated nuclei and the frequency of this abnormality was correlated with available phosphorus. The frequency of abnormalities in erythrocytes is a fish sensitive indicator of health and plays an important role as a complementary tool in the assessment of fish feeding. The variation in the frequency of nuclear abnormalities in erythrocytes obtained among fish fed with the different feeds allows the assertion that they contained genotoxic factors.

In vivo genotoxic effects of commercial grade cypermethrin on fish peripheral erythrocytes.

The study explicates the genotoxic effects of commercial grade cypermethrin on peripheral erythrocytes of Catla catla, chronically exposed to two environmentally relevant concentrations. The fish was treated with sub-lethal concentrations 0.12 μg/L and 0.41 μg/L (1/10th and /1/3rd of 96 h LC50 ) of cypermethrin for 45 days. DNA damage in the exposed fish was assessed using alkaline comet assay, presence of micronuclei (MN), erythrocyte nuclear and cytoplasmic abnormalities. Exposure to cypermethrin induced a dose-dependent increase in percent DNA damage, micronucleus frequency and erythrocyte abnormalities. Nuclear anomalies such as notched nuclei, lobed nuclei, bridged nuclei, and deformed nuclei; and cytoplasmic anomalies like anisochromasia, vacuolated cytoplasm, lobed cells, and echinocytes were observed. The findings revealed the genotoxic potential of commercial formulations pyrethroid cypermethrin at concentrations found in the environment and their potential deleterious effects on nontarget aquatic organisms.

Toxicity evaluation of the combination of emerging pollutants with polyethylene microplastics in zebrafish: Perspective study of genotoxicity, mutagenicity, and redox unbalance

Despite the toxicity of microplastics (MPs) in freshwater fish has been demonstrated in previous studies, their effects when mixed with other pollutants (organic and inorganic) are poorly understood. Thus, we aimed to test the hypothesis that the association of polyethylene MPs (PE-MPs) to a mix of emerging pollutants induces more adverse genotoxic, mutagenic, and redox unbalance effects in adult zebrafish (Danio rerio), after 15 days of exposure. Although the accumulation of MPs in animals was greater in animals exposed to PE-MPs alone, erythrocyte DNA damage (comet assay) and the frequency of erythrocytic nuclear abnormalities (ENAs) evidenced in zebrafish exposed to PE-MPs alone were as pronounced as those observed in animals exposed to the mix of pollutant (alone or in combination with MPs), which constitutes the big picture of the current study. Moreover, we noticed that such effects were associated with an imbalance between pro-and antioxidant metabolism in animals, whose activity of superoxide dismutase (SOD) and catalase (CAT) was assessed in different organs which were not sufficient to counterbalance the production of reactive oxygen species [hydrogen peroxide (H2O2)] and nitrogen [nitric oxide (NO)] evaluated. The principal component analysis (PCA) also revealed that while the antioxidant activity was more pronounced in the brain and liver of animals, the highest production of H2O2 was perceived in the gills and muscles, suggesting that the biochemical response of the animals was organ-dependent. Thus, the present study did not demonstrate antagonistic, synergistic, or additive effects on animals exposed to the combination between PE-MPs and a mix of pollutants in the zebrafish, which reinforces the theory that interactions between pollutants in aquatic ecosystems may be as complex as their effects on freshwater ichthyofauna.

Alterations in hematological parameters and the structure of peripheral erythrocytes in Nile tilapia (Oreochromis niloticus) exposed to profenofos.

The burden of pesticide use from agricultural intensification lies in the fact that pesticides may end up in aquatic ecosystems and have pernicious effects on non-target organisms, including fish. Different blood biomarkers, including hemato-biochemical indices, erythrocytic nuclear abnormalities (ENA), and erythrocytic cellular abnormalities (ECA), were observed in Nile tilapia (Oreochromis niloticus) after exposure to varying sub-lethal concentrations (0%, 5%, 10%, 20%, and 40% of 96-h LC50) of profenofos at different time intervals (7, 14, 21, and 28days). The results revealed that glucose and white blood cell (WBC) levels significantly increased, while hemoglobin, red blood cell (RBC), and packed cell volume (PCV) significantly decreased in a time- and concentration-dependent manner. Aberrant erythrocytic morphology-derived ENA, such as nuclear degeneration, micronuclear formation, binuclear development, nuclear budding, and karyopyknosis, significantly increased with time in profenofos-exposed groups compared to controls. Between the treatment and control groups, a significant execution was discerned for teardrop and fusion type ECA. For other cellular aberrations of erythrocytes, including elongated, twin, and spindle, a significant difference appeared only at the beginning of the experiment (day 7). This study concludes that the presence of widely used profenofos in aquatic systems has a pernicious effect on Nile tilapia.

Response and recovery of Nile tilapia exposed to diesel oil – Behavioral, hemato-biochemical and morphological changes of erythrocytes

Pollution caused by petroleum oil in several manners is becoming a threat to aquatic ecosystem. Hence, we carried out an experiment to investigate how diesel oil affects biota behavior, physiological attributes and how they recover by using Nile tilapia (Oreochromis niloticus) as a model organism. Nile tilapia of two different treatment groups were exposed to 0.1 mL/L and 0.5 mL/L diesel oil for 7 days. Then both groups were kept in completely diesel oil-free water for 14 days. A control group was maintained throughout the experimental period. We examined the behavioral attributes, hemato-biochemical parameters: hemoglobin (Hb), red blood cell (RBC), white blood cell (WBC) and glucose (Glu), and morphological changes of erythrocytes after diesel exposure and at the end of recovery phase. Our results revealed that there were abnormalities in behavior and significant changes in Hb, RBC, WBC and Glu level in both of the treatment group after 7 days of exposure. Frequencies of erythrocytic cellular abnormalities (ECAs), for example, twin, spindle, elongated, tear drop and erythrocytic nuclear abnormalities (ENAs) like notch nuclei, karyopyknosis, nuclear bud and nuclear bridge were prominent in both groups. However, the amount of anomalies was higher in most if not all the cases in 0.5 mL/L treatment group. Nile tilapia of both groups were quick to recover but the 0.1 mL/L group showed profound recovery than the 0.5 mL/L group. However, in the cases of ECAs and ENAs, recovery of the 0.5 mL/L group was insignificant. Hence, our experimental study concluded that the higher the exposure to diesel oil, higher incidences of major health problems are recorded, seriously piercing the healing system of Nile tilapia.