Erythrocytic Nuclear Abnormalities Research Articles

Application of fenitrothion on Heteropneustes fossilis causes alteration in morphology of erythrocytes via modifying hematological parameters

In Bangladesh, the extensive use of fenitrothion on crops and in aquaculture ponds inevitably threatens a range of aquaculture species, including fish, owing to stress responses and physiological disturbances. The present study elucidated the potential toxic effects of fenitrothion on the blood biomarkers (haemato-biochemistry and structure of erythrocytes) of stinging catfish (Heteropneustes fossilis), a commercially significant aquaculture species. Fish were exposed to four sub-lethal concentrations (0%, 10%, 20%, and 40% of the 96-h LC50 value) of fenitrothion in triplicate and observed on the 7th, 14th, 21st, and 28th day following exposure. With increasing fenitrothion concentration, blood glucose and white blood cell levels increased significantly; in contrast, hemoglobin, red blood cell, and packed cell volume substantially decreased. However, the mean corpuscular volume and mean corpuscular hemoglobin did not change significantly during the exordial period (0–7 d); although, at a later stage, changes were observed. Frequencies of observed erythrocytic nuclear abnormalities, such as degeneration, bi-nucleus, micronucleus, notch nucleus, and nuclear bridge and erythrocytic cellular abnormalities, such as echinocytes, fusion, elongation, and tear drop morphology increased significantly in a concentration-dependent manner. Differences between the control individuals and those individuals under treatment were considered insignificant for twin cells on the 14th day of exposure. The study showed the pernicious impact of the effects of fenitrothion on H. fossilis through physiological alteration, which is likely to pose challenges for aquaculture production.

Non-Clinical Investigation of Tuberculosis Drugs: Conjugated Norbornene- Based Nanocarriers Toxic Impacts on Zebrafish

INTRODUCTION: Rifampicin conjugated (R-CP), and rifampicin -isoniazid dual conjugated (RI-CP) norbornene-derived nanocarriers are newly designed for pH stimuli-responsive delivery of tuberculosis (TB) drugs. Its biosafety level is yet to be well established. OBJECTIVES: To assess the impacts of the nanocarriers on liver cells using zebrafish animal model and human liver cell line model (HepG2). METHODS: Initially, lethal dose concentration for the norbornene-derived nanocarrier systems in zebrafish was determined. The toxic effects were analysed at the sub-lethal drug concentration by histopathological study, total GSH level, gene expression and DNA damage in zebrafish liver cells. Fish erythrocyte nuclear abnormalities were also evaluated. Cell viability and oxidative stress level (ROS generation) after exposure to the nanoconjugates was determined using HepG2 cell in the in vitro study. RESULTS: In vivo studies of both R-CP and RI-CP showed 100% mortality at 96 hours for exposure concentration >100mg/l and showed toxic changes in zebrafish liver histology, GSH, and DNA damage levels. A noticeable upregulated PXR, CYP3A and cyp2p6 genes was observed in RI-CP exposure than in RIF or R-CP molecules. The in vitro study revealed a dose-dependent effect on cell viability and ROS generation for RIF, R-CP and RI-CP exposures in HepG2 cells. CONCLUSION: The current study reports that the rifampicin conjugated (R-CP) and rifampicin-isoniazid conjugated (RI-CP) norbornene derived nanocarriers exhibit enhanced toxic responses in both adult zebrafish and HepG2 cells. The pH-sensitive norbornene derived nanocarriers on conjugation with different drugs exhibited varied impacts on hepatic cells. Hence the present investigation recommends a complete metabolomics analysis and norbornene carrier-drug interaction study to be performed for each drug conjugated norbornene nanocarrier to ensure its biosafety.

Genotoxicity, oxidative stress and lysozyme induction in Clarias gariepinus chronically exposed to water-soluble fraction of burnt tire ash.

The safety of aquatic ecosystems has been compromised by numerous anthropogenic activities, especially leachates from non-point source toxicants, leaching into aquatic systems. This study evaluated the toxicity of the water-soluble fractions (WSFs) of burnt tire ash (BTA) on Clarias gariepinus via a battery of integrated biomarkers. Juvenile C. gariepinus were exposed to sublethal (0.56, 1.12, and 2.24 g/L) concentrations of BTA, derived from 11.2 g/L median lethal concentration (96 LC50), at duration intervals of 1, 14, and 28 days, followed by a recovery trial that lasted for 14 days. Serum biochemical parameters, antioxidant enzyme activities of the gill and liver, lysozymes activity and erythron profile were assessed. The findings of the present study revealed that BTA-WSF induced prominent alterations on biochemical parameters, lysozymes activity and antioxidant enzymes activities in the exposed fish. Furthermore, toxicant exposure promoted oxidative stress, cellular damage and genotoxicity (erythrocytic nuclear and cellular abnormalities) in the exposed fish. In general, a post-exposure trial showed partial recovery from the exposure effects of the toxicant, following the evident modifications of serum enzymes and erythron pathopathology in the experimental model. Biomonitoring of BTA, using sentinel aquatic species such as C. gariepinus, provides insights into the ecotoxicological potency of this toxicant.

Habitat differences affect the nuclear morphology of the erythrocytes and the hepatic melanin in Leptodactylus fuscus (Anura) in the Brazilian Cerrado savanna.

The sensitivity of anuran to the effects of habitat destruction and contamination has led to a preoccupying global decline in their populations. Morphological biomarkers such as micronuclei and other erythrocyte nuclear abnormalities (ENAs), as well as the occurrence of hepatic melanin, can be used to evaluate the effects of habitat impacts. In the present study, these two parameters were combined for the in situ assessment of the effects of soybean cultivation on the grassfrog, Leptodactylus fuscus. Specimens were also collected from a protected area to provide a reference site (non-agricultural environment). The frequency of some of the nuclear abnormalities in the animals from the soybean plantation was much higher than that recorded at the reference site, in particular micronuclei, which were 3.6 times more frequent in the plantation, lobulated nuclei (3.4 times more frequent), and reniform nuclei, which were four times more common than at the reference site. The combined analysis of all the ENAs together also revealed a frequency approximately 1.4 times higher in the animals from the soybean plantation, in comparison with the protected area. Smaller areas of hepatic melanin were observed in the specimens from the soybean plantation. These results provide further evidence of the sensitivity of anurans to habitat impacts and indicate that animals found in soybean plantations are susceptible to systematic alterations of their cells.

Impacts of wildfires in aquatic organisms: biomarker responses and erythrocyte nuclear abnormalities in Gambusia holbrooki exposed in situ.

Wildfires are an environmental concern due to the loss of forest area and biodiversity, but also because their role as drivers of freshwater systems contamination by metals. In this context, the fish Gambusia holbrooki was used as a model, deployed for in situ exposure in watercourses standing within a recently burnt area and further assessment of toxic effects. The fish were exposed during 4 days at four different sites: one upstream and another downstream the burnt area and two within the burnt area. Biochemical biomarkers for oxidative stress and damage were assessed. The extent of lipoperoxidative damage was monitored by quantifying malondialdehyde and DNA damage evaluated through erythrocyte nuclear abnormalities observation. Chemical analysis revealed higher metal levels within the burnt area, and exposed fish consistently showed pro-oxidative responses therein, particularly an increase of gill glutathione peroxidase and glutathione reductase activity, the records doubling compared to samples from sites in the unburnt area; also the activity of glutathione-S-transferases comparatively increased (by 2-fold in the liver) in samples from the burnt area, and malondialdehyde was produced twice as much therein and in samples downstream the burnt area reflecting oxidative damage. Consistently, the frequency of erythrocyte nuclear abnormalities was higher at sites within and downstream the burnt area. This study supports the use of sensitive oxidative stress and genotoxicity biomarkers for an early detection of potentially noxious ecological effects of wildfires runoff.

Multiple toxicity endpoints induced by carbon nanofibers in Amazon turtle juveniles: Outspreading warns about toxicological risks to reptiles

The toxicity of carbon-based nanomaterials (CNs) has been observed in different organisms; however, little is known about the impact of water polluted with carbon nanofibers (CNFs) on reptiles. Thus, the aim of the current study was to assess the chronic effects (7.5months) of 1 and 10mg/L of CNF on Podocnemis expansa (Amazon turtle) juveniles (4months old) based on different biomarkers. Increased total organic carbon (TOC) concentrations observed in the liver and brain (which suggests CNF uptake) were closely correlated to changes in REDOX systems of turtles exposed to CNFs, mainly to higher nitrite, hydrogen peroxide and lipid peroxidation levels. Increased levels of antioxidants such as total glutathione, catalase and superoxide dismutase in the exposed animals were also observed. The uptake of CNFs and the observed biochemical changes were associated with higher frequency of erythrocyte nuclear abnormalities (assessed through micronucleus assays), as well as with both damage in erythrocyte DNA (assessed through comet assays) and higher apoptosis and necrosis rates in erythrocytes of exposed turtles. Cerebral and hepatic acetylcholinesterase (AChE) increased in turtles exposed to CNFs, and this finding suggested the neurotoxic effect of these nanomaterials. Data in the current study reinforced the toxic potential of CNFs and evidenced the biochemical, mutagenic, genotoxic, cytotoxic, and neurotoxic effects of CNFs on P. expansa.

Can the insecticide Imidacloprid affect the health of the Neotropical freshwater fish Astyanax altiparanae (Teleostei: Characidae)?

Female juveniles of the Neotropical fish Astyanax altiparanae were exposed for 96 h to four treatments containing the active ingredient from Imidacloprid® commercial formulation (IMI 1, IMI 2, IMI 3, and IMI 4) and to a control treatment (only dechlorinated tap water). Glutathione content, glutathione S-transferase activity, lipid peroxidation (LPO) and protein carbonylation levels, acetylcholinesterase (AChE) activity, and frequency of micronuclei and erythrocyte nuclear abnormalities (ENA) were measured in the fish. The muscle and gills were the most affected organs; their antioxidant defense was not enough to prevent oxidative damage (LPO) in the IMI 2 and IMI 4 treatment fish. IMI also inhibited AChE activity in the muscle (IMI 3 and IMI 4) and increased ENA frequency (IMI 4). IMI can affect the health of A. altiparanae in environmentally relevant concentrations, causing oxidative damage in different organs, neurotoxic effects in the muscle, and genotoxicity.

Giant Toads (Rhinella marina) From the Industrial Zones of Low Basin of the Coatzacoalcos River (Veracruz, MX) Presents Genotoxicity in Erythrocytes.

The lower basin of Coatzacoalcos River is one of the most polluted regions of the southern Gulf of Mexico. Organochlorine compounds, polybrominated diphenyl ethers, polycyclic aromatic hydrocarbons, and heavy metals have been registered in this region. In the present study, genotoxicity was evaluated in the blood of giant toads (Rhinella marina) from Coatzacoalcos’ rural and industrial zones, and compared with laboratory toads. Determination of the frequency of micronucleus and erythrocyte nuclear abnormalities by the light microscope and cell cycle and apoptosis by flow cytometry were used as biomarkers of genotoxicity. We found more variability in micronucleus and more nuclear buds in toads from industrial zones. Also, cell cycle alterations and an increase of apoptosis in erythrocytes were found in toads from rural and industrial zones. Multivariate statistics show that the toads from the industrial zone were more affected than toads from laboratory and rural zones.Supplementary InformationThe online version contains supplementary material available at 10.1007/s00128-021-03162-2.

Bisphenol A and Bisphenol S Induce Endocrine and Chromosomal Alterations in Brown Trout.

Bisphenol A is a widely used compound found in large amount of consumer products. As concerns have been raised about its toxicological and public health effect, the use of alternatives to bisphenol A are now increasing. Bisphenol S is one of the analogues being used as a replacement for bisphenol A despite the fact that little is known about the effects of bisphenol S on living organisms. In this study, we investigated the potential endocrine and genotoxic effects of bisphenol A and bisphenol S in juvenile brown trout (Salmo trutta). The fish were exposed to the compounds for either 2 weeks or 8 weeks via sustained-release cholesterol implants containing doses of 2 mg/kg fish or 20 mg/kg fish of the substances. The effects on the thyroid hormone levels and the estrogenic disrupting marker vitellogenin were evaluated, along with the genotoxic markers micronucleated cells and erythrocyte nuclear abnormalities. An increase in plasma vitellogenin was observed in fish exposed to the high dose of bisphenol A for 2 weeks. At this experimental time the level of the thyroid hormone triiodothyronine (T3) in plasma was elevated after bisphenol S exposure at the high concentration, and paralleled by an increase of micronucleated cells. Moreover, bisphenol A induced an increase of micronuclei frequency in fish erythrocytes after the exposure at the lowest dose tested. Taken together the results indicate that both bisphenol A and its alternative bisphenol S cause endocrine disrupting and genotoxic effects in brown trout, although suggesting two different mechanisms of damage underlying bisphenol A and bisphenol S activity.

DNA damage induced by cylindrospermopsin on different tissues of the biomonitor fish Poecilia reticulata.

The cyanotoxin cylindrospermopsin (CYN) is the second biggest cause of poisoning worldwide, both in humans and animals. Although CYN primarily affects the aquatic environments and can be absorbed in fishes by multiple routes, data reporting its toxicity and mechanism of action are still scarce in this group. Using P. reticulata as model species, it was evaluated whether CYN promotes mutagenic and genotoxic effects in different fish target tissues. Adult females were exposed in a static way to 0 (control), 0.5, 1.0, and 1.5 μg L-1 of pure CYN for 24 and 96 hours. For the first time, DNA damage was detected in fish brain after CYN exposition. In brain cells, a concentration-response DNA damage was observed for both exposure times, suggesting a direct or indirect action of CYN in neurotoxicity. For the liver cells, 96 hours caused an increase in DNA damage, as well the highest percentage of DNA in the tail was reached when used 1.5 μg L-1 of CYN. In peripheral blood cells, an increase in DNA damage was observed for all tested concentrations after 96 hours. In erythrocytes, micronuclei frequency was higher at 1.5 μg L-1 treatment while the erythrocyte nuclear abnormalities (ENA) frequency was significantly higher even at the lowest CYN concentration. Such data demonstrated that acute exposition to CYN promotes genotoxicity in the brain, liver, and blood cells of P. reticulata, as well mutagenicity in erythrocytes. It rises an alert regarding to the toxic effects of CYN for aquatic organisms as well as for human health.

Exposure to a contaminated tropical freshwater (Awba Dam) in Ibadan, Nigeria, induced cytogenotoxicity and haemato-pathological changes in Clarias gariepinus.

Awba reservoir serves the purpose of water supply in the University of Ibadan, Nigeria. Recent reports on pollution status have focused on toxicological implication of contaminants in this reservoir. But none is on genetic and systemic toxicity of the water in fish. We investigated cytogenotoxicity of Awba Dam water (ADW) on Clarias gariepinus using piscine micronucleus (MN) assay. Haematological and histopathological changes were also evaluated. Bi-monthly composite water samples were collected from the reservoir from July to October, 2018. The water was used to cultivate juvenile C. gariepinus in the laboratory for 1-4 months, and tap water was used as the negative control. Peripheral blood erythrocytes from the caudal veins of C. gariepinus were used for the monthly MN assessment. There was significant (p < 0.05) induction of MN and other erythrocyte nuclear abnormalities in C. gariepinus within the period of study. There were variations in the haematological indices and pathological alterations in sections of the gill, liver and kidney of C. gariepinus. The levels of some heavy metals in ADW were above standard permissible limits and might have contributed to the observed cytogenetic and systemic disruptions. These findings may be used by the concerned authorities to evolve management strategies for the reservoir's health and biologic resources.

CYTOGENETIC ANALYSIS OF NUCLEAR ANOMALIES OF ERITHROCYTES OF COMMON CARP (CYPRINUS CARPIO)

Blood of fish is a highly sensitive system that quickly responds to the effects of toxins and other stress factors of abiotic and biotic nature. Among the various pathologies of fish blood the formation of micronuclei and anomalies of the nucleus of erythrocytes are distinguished, which can act as markers of the stability of the genome of aquatic life. Therefore, this work presents a cytogenetic analysis of nuclear abnormalities in erythrocytes of common carp. In fish under investigation, erythrocytes with micronuclei, with a budding and bubbling nucleus, erythrocytes with a caudate filamentous and coracoid nucleus, and cells with a bilobed nucleus were detected. In the blood such types of nucleus anomalies as erythrocytes with a budding and bubbling nucleus prevailed, constituting 0.16 ± 0.031 and 0.28 ± 0.153%, respectively. The average total level of pathologies of the nucleus of erythrocytes in the studied carp was 0.89 ± 0.202%, which corresponded to the parameters of healthy fish.

Distortion of micronuclei and other peripheral erythrocytes caused by fenitrothion and their recovery assemblage in zebrafish.

The experiment was explicated to investigate the fenitrothion persuaded genotoxicity in the peripheral erythrocytes of zebrafish (Danio rerio) through in vivo exposures (10 %, 20 % and 40 % of LC50 of fenitrothion, i.e., 0.8, 1.6, and 3.2 mg/L, respectively) for variable periods (1, 3, and 7 days) and its subsequent post-exposure recuperation array in pesticide-free water for similar intervals was also evaluated. With the exception of the control group (0% of fenitrothion), the obtained results pointed out that with the promotion of time and concentrations, fenitrothion induced significantly (p < 0.05) higher prevalence and severity of erythrocytic nuclear abnormalities (ENA) such as- notched, micronucleus, nuclear bridges, blebbed, binucleated, nuclear bud and also erythrocytic cellular abnormalities (ECA) such as - echinocytic, elongated, tear-drop, crescentic, twin, fusion, and spindle-shaped cells. Recuperation data stated that zebrafish cured spontaneously and aberrated erythrocytic anomalies in all treatments were renormalized according to the concentration and duration dependence. Hence, we concluded that fenitrothion has a dangerous effect on the zebrafish, and this technology can be used to anticipate the sensitivity of aquatic animals to environmental pollution.

Influence of extreme ambient cold stress on growth, hematological, antioxidants, and immune responses in European seabass, Dicentrarchus labrax acclimatized at different salinities

Climate change-driven extreme climatic events are expected to challenge ectotherms' physiological tolerance. The hemato-physiological modulation potentials of fish during ambient extreme-cold events at various salinities are poorly studied. In this study, we evaluated the growth, hemato-physiological, antioxidants, and immunological responses of European seabass, Dicentrarchus labrax acclimatized at 30, 12, 6, and 3 PSU followed by an extreme ambient cold (8 °C) exposure for 20 days. Juveniles acclimatized at 30 and 3 PSU showed significantly low growth performance (p < 0.05). Red blood cells (RBC) count, hematocrit, hemoglobin, and serum protein content were decreased in 3 and 30 PSU fish. In contrast, significantly higher white blood cells (WBC) count, skin mucus cortisol, different types of erythrocytic cellular abnormalities (ECA), and erythrocytic nuclear abnormalities (ENA) were observed in 30 and 3 PSU fish. Also, higher activities of serum antioxidants [superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT), glutathione reductase (GR)], and upregulation of tumor necrosis factor α (TNF-α) in the spleen were observed in 3 and 30 PSU fish. In addition, on day 20, phagocytic respiratory burst (RB) and serum lysozyme activities (LSZ) were significantly higher in 3 and 30 PSU fish during extreme cold exposure. None of the repeatedly evaluated parameters indicated acclimation capacity to cope with tested salinities during cold exposure. However, taken together, our results indicate that Dicentrarchus labrax acclimatized at intermediary salinities (6 and 12 PSU) can perform comparatively better during ambient extreme-cold exposure (8 °C).

Chromium Exposure Causes Structural Aberrations of Erythrocytes, Gills, Liver, Kidney, and Genetic Damage in Striped Catfish Pangasianodon hypophthalmus.

Heavy metal pollution due to anthropogenic activities poses a great threat to aquatic organisms. The present study was conducted to evaluate the cytotoxic and genotoxic effects of hexavalent chromium (potassium dichromate) on hemato-biochemical, histo-pathological, and genetical changes in striped catfish Pangasianodon hypophthalmus. Three sub-lethal doses (0.8, 1.6, and 3.2 mg/L) of chromium (Cr) were selected and fish were exposed in vivo contrasting with a control (0 mg/L) for 30 days. The study revealed that various hemato-biochemical parameters showed a significant decrease in hemoglobin (Hb), red blood cells (RBCs), and blood glucose content, whereas white blood cells (WBCs) significantly increased in Cr exposed fish. Frequencies of all forms of structural abnormalities of erythrocytes (erythrocytic cellular abnormalities; ECA, erythrocytic nuclear abnormalities; ENA and erythroblasts; Ebs) were significantly increased in higher two test concentrations (1.6 and 3.2 mg/L) when compared to control. Differential leucocyte count exhibited significant increase in neutrophil and decrease in lymphocytes in the highest Cr treated group. The severity of various histo-pathological changes in the gills, liver, and kidney were increased considerably with the increase of Cr concentrations. Similarly, the amount of DNA (ng/μl) decreased significantly in blood and tissues of different vital organs where the liver showed the highest decline compared to control in a concentration-dependent manner. Taken altogether, P. hypophthalmus is susceptible to Cr and can be used as a bio-indicator to assess aquatic metal pollution.

Co-exposure of iron oxide nanoparticles and glyphosate-based herbicide induces DNA damage and mutagenic effects in the guppy (Poecilia reticulata)

Iron oxide nanoparticles (IONPs) have been tested to remediate aquatic environments polluted by chemicals, such as pesticides. However, their interactive effects on aquatic organisms remain unknown. This study aimed to investigate the genotoxicity and mutagenicity of co-exposure of IONPs (γ-Fe2O3 NPs) and glyphosate-based herbicide (GBH) in the fish Poecilia reticulata. Thus, fish were exposed to citrate-functionalized γ-Fe2O3 NPs (0.3 mg L−1; 5.44 nm) alone or co-exposed to γ-Fe2O3 NPs (0.3 mg L−1) and GBH (65 and 130 μg of glyphosate L−1) during 14 and 21 days. The genotoxicity (DNA damage) was analyzed by comet assay, while the mutagenicity evaluated by micronucleus test (MN test) and erythrocyte nuclear abnormalities (ENA) frequency. The co-exposure induced clastogenic (DNA damage) and aneugenic (nuclear alterations) effects on guppies in a time-dependent pattern. Fish co-exposed to NPs and GBH (130 μg glyphosate L−1) showed high DNA damage when compared to NPs alone and control group, indicating synergic effects after 21 days of exposure. However, mutagenic effects (ENA) were observed in the exposure groups after 14 and 21 days. Results showed the potential genotoxic and mutagenic effects of maghemite NPs and GBH co-exposure to freshwater fish. The transformation and interaction of iron oxide nanoparticles with other pollutants, as herbicides, in the aquatic systems are critical factors in the environmental risk assessment of metal-based NPs.

Genotoxic effects of glyphosate on Physalaemus tadpoles

Genotoxicity studies have revealed that pesticides bind to genetic material in non-target vertebrates, thereby impairing the genetic integrity of these animals. The main objective of this study was to determine the genotoxic damage in erythrocytes of two native South American amphibian Physalaemus cuvieri and Physalaemus gracilis, both species exposed to a glyphosate-based herbicide. We evaluated the presence of micronuclei (MN) and erythrocyte nuclear abnormalities (ENA) as biomarkers for potential genotoxic compounds. Tadpoles were exposed to doses permitted by Brazilian legislation and concentrations found naturally in Brazilian and Argentinian waters (500, 700 and 1000 μg/L). Glyphosate-based herbicide caused micronuclei formation and several types of erythrocyte nuclear abnormalities in both Physalaemus species. The total frequency of MN and ENA demonstrated the occurrence of cell damage at all tested concentrations. Glyphosate herbicide can be considered a genotoxic that may impact the genetic integrity of native populations of P. cuvieri and P. gracilis.

Multi-biomarkers approach to access the impact of novel metal-insecticide based on flavonoid hesperidin on fish.

Aquatic ecosystem health is the main concern to increasing pesticides application to control agricultural pests as it is the ultimate receptor of such materials. This study evaluated the impact of new metal-insecticide, the [Mg(hesp)2(phen)], referred as MgHP, on fish using physiological, genetic, biochemical, and morphological biomarkers. The fish, Prochilodus lineatus, was exposed to 0 (control), 1, 10, 100, 1000μgL-1 MgHP, for 24 and 96h. MgHP was not lethal but caused genotoxicity, altered hematological variables and, the activity of antioxidant and biotransformation enzymes and histology of liver, depending on concentration and time exposure. Hematocrit and erythrocyte number (RBC) increased without change hemoglobin content resulting in changes in hematimetric indexes after 24h; after 96h, only RBC was changed. Erythrocyte nuclear abnormalities and crenate cells increased after 24h but, not after 96h. Erythrocytes and hepatocytes indicated instability in DNA integrity however, the absence of micronuclei suggested DNA damage repairment. After 24h, the antioxidant defense system and the phase II biotransformation enzyme was responsiveness and catalase activity decreased at high MgHP concentrations; the antioxidant response was triggered after 96h. Hepatocyte hypertrophy, intracellular cytoplasmic substances, cytoplasm degeneration, melanomacrophage and hyperemia increased in fish exposed from 10μgL-1 to higher MgHP concentrations; the organ alteration index increased as MgHP concentration increased showing dose-dependence. Most of hematological and genotoxic effects occurred after 24h exposure evidencing potential recover capability of organism by activation of the antioxidant defense system and DNA repairment mechanisms. Nevertheless, the histopathological changes in the liver was maintained over time at high MgHP concentrations, a concentration usually no environmental relevant. In conclusion, this data reinforced the importance of continuing research on MgHP effects in other organisms considering the promising use of such compound to control the leaf-cutter ants and other insects.

High temperature acclimation alters upper thermal limits and growth performance of Indian major carp, rohu, Labeo rohita (Hamilton, 1822).

Increase in water temperature due to anthropogenic and climatic changes is expected to affect physiological functions of fish. In this study, we determined high temperature tolerance (CTmax) of a common aquacultured Indian major carp, rohu, Labeo rohita fingerlings (15.96±0.72g BW, 11.56±0.42cmTL) followed by acclimatization at three temperatures (30, 33, 36°C). To determine the CTmax, we analyzed the major hemato-biochemical indices - hemoglobin (Hb), red blood cell (RBC), white blood cell (WBC), blood glucose levels, and erythrocytic nuclear abnormalities (ENAs) and erythrocytic cellular abnormalities (ECAs) of peripheral erythrocytes in the fish sampled at the start and end point at each acclimated temperature. Significantly decreased CTmax of the fish was found at 36°C compared to 30°C and 33°C. The fish in the highest (36°C) temperature were found with significantly lower Hb and RBC content and significantly higher WBC and blood glucose levels than that of the fishes in the lowest (30°C) temperature both at the start and end points. The highest frequencies of ENAs and ECAs were found in the highest (36°C) temperature group compared to the lowest (30°C) temperature group at both the points. We also evaluated growth performance of the rohu fingerlings reared in the three temperatures for 60 days. The growth parameters - final weight gain, percent weight gain and specific growth rate were the highest at 33°C and the lowest at 36°C. The present study revealed that the highest temperature (36°C) tested here may be hazardous to rohu and the temperature should be kept below 36°C in the aquaculture setting to avoid physiological damage and growth and production loss to the fish.

Histopathologic and genotoxic effects of deltamethrin on marsh frog, Pelophylax ridibundus (Anura: Ranidae)

It is known that amphibians inhabiting agricultural areas are constantly exposed to chemicals such as insecticides. Deltamethrin, a type II pyrethroid insecticide, is widely used in the world. The present study aimed to investigate the toxic effects of different concentrations (0.625, 1.25, and 2.50 mg/kg body wt) of orally applied deltamethrin to marsh frog (Pelophylax ridibundus) after 96 h of treatment. The histopathological abnormalities in liver, gastrointestinal tract, and kidney tissues were evaluated with quantitative analyses. In liver, increase of melanomacrophagic aggregates, non-homogeneous hepatocyte parenchyma, sinusoidal dilatations, infiltration, vascular epithelial degeneration, central vein degeneration, and congestion were determined in exposed frogs. In gastrointestinal tract, vacuolization, hypertrophy, congestion, infiltration, necrosis, and erosion of the epithelial layer, increasing goblet secretion, degeneration in villi, epithelial disorganization, and edema were observed in high-dose groups. In all experimental groups, glomerular shrinkage, hemorrhage, degeneration, infiltration, increase in Bowman distance, and eosinophilic-stained tubular lumens were detected in kidneys. Histopathological changes were more prominent in 1.25 and 2.50 mg/kg groups than the other groups. To determine the genotoxic effects of deltamethrin, the peripheral blood samples of the frogs were used. The erythrocytic nuclear abnormalities such as micro-nucleus, double-nucleus, kidney-shaped nucleus, notched nucleus, and bud nucleus were determined in the frogs, and the genotoxicity did not show a significant increase between control and low-dose groups, statistically. However, the genotoxic effects increased in medium- and especially high-dose groups. The results of this study showed that acute deltamethrin exposure can lead to histopathologic and genotoxic effects in P. ridibundus.