Mixture of pesticides based on dimethylamine and imidacloprid affects locomotion of adult zebrafish.

Numerous chemical compounds are found in aquatic environments; among them are pesticides. Pesticides are widely used worldwide, and this use has progressively increased in recent decades, resulting in the accumulation of potentially toxic compounds in surface waters. Dimethylamine-based herbicides (DBH) and imidacloprid-based insecticides (IBI) have low soil absorption and high water solubility, facilitating the arrival of these compounds in aquatic environments. In this study, our objective was to analyze whether two pesticides, DBH and IBI at environmentally relevant concentrations of 320 µg/L for each compound, and their mixtures impact the behavioral and endocrine parameters of adult zebrafish, verifying the effect of pesticides on exploratory behavior and social and analyzing hormonal parameters related to stress. Acute exposure to the mixture of pesticides reduced fish locomotion. Pesticides alone and in combination did not affect cortisol levels in exposed animals. Pesticides, when tested together, can cause different effects on non-target organisms, and the evaluation of mixtures of these compounds is extremely important.

Cannabis sativa-based oils against aluminum-induced neurotoxicity.

The use of terpenoid compounds in different neural-related conditions is becoming useful for several illnesses. Another possible activity of these compounds is the reduction of nervous impairment. Cannabis sativa plants are known for their concentration of two important terpenoids, the delta-9-tetrahydrocannabinol (THC) and cannabidiol (CBD). CBD and THC have central peripheral activities already described and their usage in different brain diseases, such as Alzheimer's and multiple sclerosis. Aluminum (Al) is known as an important neurotoxic compound, the physiological action of Al is not known already, and in high concentrations can lead to intoxication and cause neurotoxicity. Here we evaluated the potential effect of two different doses of CBD- and THC-rich based oils against Al-induced toxicity, in the zebrafish model. We evaluated behavioral biomarkers of the novel tank test (NTT) and social preference test (SPT), and biochemical markers: the activity of the enzyme acetylcholinesterase (AChE) and the antioxidant enzymes-catalase, superoxide dismutase, and glutathione-S-transferase. CBD- and THC-based oils were able to increase the AChE activity helping the cholinergic nervous system actuate against Al toxicity which was reflected by the behavioral biomarkers changes. We concluded that the oils have a protective effect and might be used with proposals for neurological and antioxidant impairment avoidance caused by Al intoxications.

Pyrethroid-based insecticides exert transgenerational, persistent, and chronic effects in Caenorhabditis elegans.

The use of agrichemical pyrethroid-based insecticides to combat crop infestations and ectoparasites in animals is increasing. In this context, two pyrethroid-based insecticides are widely used, λ-cyhalothrin and Cypermethrin. The mechanism of action of these insecticides is characterized by the opening of ion channels and death by neural hyperexcitability. In this study, we evaluated the toxicological effect of two pyrethroid-based insecticides λ-cyhalothrin and Cypermethrin in C. elegans aiming to evaluate the transgenerational (TG), neonatal (NN) and lifespan (LS) effects of these compounds. At the end of each exposure period, were evaluated behavioral biomarkers of body bends, pharyngeal pumping, and feeding behavior. Furthermore, the fluorescent expression of antioxidant enzymes (superoxide dismutase, catalase, and glutathione-S-transferase) and the fluorescent expression of PolyQ40 aggregates were quantified. Lastly, the activity of the enzyme acetylcholinesterase (AChE) was quantified. Changes in TG were more related to changes in AChE enzyme activity that probably were transferred to the offspring, altering behavioral biomarkers in the adult life of offspring from exposed parents. However, alterations in LS were related to the modulation of ion channels in a chronic way, exerting behavioral effects. In addition, both compounds increased the expression of PolyQ40 muscle aggregates in mutant worms. These proteins are related to the increased probability of the senile incidence of Huntington's Disease in genetically predisposed patients.

Neurotoxicological effects of venlafaxine on Caenorhabditis elegans and Danio rerio.

The growing consumption of psychoactive drugs, such as Venlafaxine (VFX), can negatively affect the organisms. Our main hypothesis is to investigate if VFX at human-used doses could exert effects on the behavioral, nervous, and antioxidant systems of two different organisms, zebrafish and C. elegans. We evaluated the effect of acute exposure to VFX at four concentrations (0, 37.5, 75, and 150 mg L-1) using toxicological indicator assessments. We evaluated zebrafish behavior using the novel tank test (NTT), social preference test (SPT), cortisol levels, acetylcholinesterase (AChE) activity, and antioxidant system. In C. elegans, we evaluated body bends, defecation cycles, pharyngeal pumping, AChE activity, and antioxidant system. C. elegans do not show alterations in the behavior analysis of pharyngeal pumping and body bends. Instead, the defecation cycle was increased in the highest dose of VFX. AChE activity also does not have differences compared to the control, the same occurs in lipid peroxidation rates. These results showed that nematodes were more resistant to changes when exposed to VFX. Zebrafish exposed to VFX showed changes in the NTT and SPT test, mainly in the anxiolytic pattern, suggesting that VFX alters this anxiolytic-like behavior. Comparing both organisms, we can observe that zebrafish seems to be more sensitive in this neurotoxicological evaluation.

Behavioral, biochemical, and endocrine responses of zebrafish to 30-min exposure with environmentally relevant concentrations of imidacloprid-based insecticide.

The imidacloprid-based insecticides (IBIs) are among the most used insecticides worldwide, and chronic and acute toxic effects (days exposure protocols) have been reported in several species in studies of IBIs at lethal concentrations. However, there is little information on shorter time exposures and environmentally relevant concentrations. In this study, we investigated the effect of a 30-min exposure to environmentally relevant concentrations of IBI on the behavior, redox status, and cortisol levels of zebrafish. We showed that the IBI decreased fish locomotion and social and aggressive behaviors and induced an anxiolytic-like behavior. Furthermore, IBI increased cortisol levels and protein carbonylation and decreased nitric oxide levels. These changes were mostly observed at 0.013 and 0.0013 µg·L-1 of IBI. In an environmental context, these behavioral and physiological disbalances, which were immediately triggered by IBI, can impair the ability of fish to evade predators and, consequently, affect their survival.

A novel insight on SARS-CoV-2 S-derived fragments in the control of the host immunity.

Despite all efforts to combat the pandemic of COVID-19, we are still living with high numbers of infected persons, an overburdened health care system, and the lack of an effective and definitive treatment. Understanding the pathophysiology of the disease is crucial for the development of new technologies and therapies for the best clinical management of patients. Since the manipulation of the whole virus requires a structure with an adequate level of biosafety, the development of alternative technologies, such as the synthesis of peptides from viral proteins, is a possible solution to circumvent this problem. In addition, the use and validation of animal models is of extreme importance to screen new drugs and to compress the organism's response to the disease. Peptides derived from recombinant S protein from SARS-CoV-2 were synthesized and validated by in silico, in vitro and in vivo methodologies. Macrophages and neutrophils were challenged with the peptides and the production of inflammatory mediators and activation profile were evaluated. These peptides were also inoculated into the swim bladder of transgenic zebrafish larvae at 6 days post fertilization (dpf) to mimic the inflammatory process triggered by the virus, which was evaluated by confocal microscopy. In addition, toxicity and oxidative stress assays were also developed. In silico and molecular dynamics assays revealed that the peptides bind to the ACE2 receptor stably and interact with receptors and adhesion molecules, such as MHC and TCR, from humans and zebrafish. Macrophages stimulated with one of the peptides showed increased production of NO, TNF-α and CXCL2. Inoculation of the peptides in zebrafish larvae triggered an inflammatory process marked by macrophage recruitment and increased mortality, as well as histopathological changes, similarly to what is observed in individuals with COVID-19. The use of peptides is a valuable alternative for the study of host immune response in the context of COVID-19. The use of zebrafish as an animal model also proved to be appropriate and effective in evaluating the inflammatory process, comparable to humans.

Tank color influences the response of tomato clownfish (Amphiprion frenatus) to an acute stress challenge.

The trade of aquarium organisms is growing worldwide. This market depends on a continuous supply of healthy and colorful aquatic animals, but this sector has few initiatives. However, in the last decade, there has been a growing interest in researching captive breeding of these animals, aiming to develop a more sustainable aquarium hobby. Larviculture is an important phase in the cultivation process because the larvae are more sensitive to stress and variations in the bulk of variables, such as temperature, salinity, nutritional management, light intensity and spectrum, and environmental background colors. Because background color could be a promoter variable of proper welfare, we tested whether it affects the endocrine response of tomato clownfish Amphiprion frenatus larvae to an acute stress challenge. We show that background color influences the endocrine stress axis responsiveness in tomato clownfish. When fish were subjected to a standard acute stressor of 61 days after hatching, only fish adapted to white walls increased the whole-body cortisol levels. From the results presented herein, we recommend that white tanks be avoided for A. frenatus larviculture. Both, the less stress level and the good welfare condition of larvae reared in colored tanks may have robust, practical applications since almost all clownfish in the ornamental aquarium trade come from captive breeding.

Household based-pyrethroids on adult zebrafish (Danio rerio) exert behavioral and cholinergic changes in different brain regions.

Pyrethroid-based insecticides are largely used for mosquito control. These compounds have household and agricultural applications with different formulations. Two important compounds used as household insecticides are prallethrin and transfluthrin, both from the pyrethroid chemical group. With the mode of action centered on sodium channels, pyrethroids keep the ionic sodium channels open for a long time causing the death of the insect by nervous hyperexcitability. Given the increased use of household insecticides by humans and the incidence of disease outbreaks with unknown etiology such as autism spectrum disease, schizophrenia, and Parkinson's disease we investigate some physiological inputs of these compounds on zebrafish. In this study, we evaluated the social interaction, shoaling formation, and anxiety-like behavior of zebrafish exposed chronically to transfluthrin- and prallthrin-based insecticides (T-BI and P-BI). In addition, we quantified the activity of the enzyme acetylcholinesterase (AChE) in different brain regions. We observed that both compounds caused anxiolytic behavior and reduced shoaling formation and social interaction. Their behavioral biomarkers indicated a harmful ecological effect on the specie as well as a possible impact of these compounds on autism spectrum disorder (ASD) and schizophrenia (SZP). In addition, the AChE activity would change its activity in different brain regions modulating the anxiety-like behavior and social behavior in zebrafish. We conclude that P-BI and T-BI make us alert about the relationship of these compounds with nervous diseases related to cholinergic signaling.

Effects of used and under-used doses of Transfluthrin-based insecticide on Caenorhabditis elegans metabolism.

Different classes of insecticide compounds have been employed to control insects and mosquitoes; Pyrethroids are one of the most common used in both urban and rural household environments. This study investigated the effects of exposure of two doses of commercial transfluthrin-based insecticide (T-BI) on behavior (body bends, pharyngeal pumping rate, and feeding attributes) and biochemical biomarkers (AChE, PolyQ40 aggregations, HSP, antioxidative SOD, CTL, and GST) following three different protocols (transgenerational, neonatal, and lifespan) in Caenorhabditis elegans model system. The relative calculated dose (RCD) and relative calculated half dose (RCHD) of T-BI were compared with those of the control (water). T-BI reduced the health span of worms treated during their whole life and changed biochemical and behavioral patterns due to progenitors' uterine (transgenerational) and neonatal exposures. It was inferred that the effects of T-BI are transgenerational and persistent and can be harmful to non-target species, including humans. In addition, our findings highlight that T-BI contact by progenitors accelerates the establishment of Huntington's disease and causes a cholinergic outbreak in offspring adulthood.

Exposure to levonorgestrel-based birth control pill in early life and its persistent effects in zebrafish.

The consumption of progestins has increased considerably in recent decades, as has their disposal into the environment. These substances can negatively affect the reproduction, physiology, and behavior of non-target organisms, such as fish. We aimed to evaluate the effects of exposure to environmentally relevant concentrations of levonorgestrel-control birth based (1.3, 13.3, 133, and 1330 ng/L) on the development and behavior of zebrafish (Danio rerio) in terms of mortality, hatching, spontaneous movement, and larval and adult behavioral tests. Exposure caused anxiogenic-like behavior in larvae, which persisted in adults, as demonstrated by the light-dark test. In contrast, it caused anxiolytic-like behavior in the novel tank test. There was a high mortality rate at all tested concentrations and increases in the hormone cortisol at 13.3 ng/L that affected the sex ratio. These changes may lead to an ecological imbalance, emphasizing the risk of early exposure to progestins in the environment.

Household prallethrin-based insecticide toxicity on different C. elegans life stage: A possible sign of Huntington Disease.

Household insecticide is largely used for insect and ectoparasite control, in city centers as well as in the countryside. The pyrethroids are the most used class of insecticide, these compounds in low doses have low toxicity for mammalians, in comparison to other compounds, with insecticide effects. The contact of these compounds in sublethal doses begins in early life and many cases, in intrauterine life. Considerable diseases still with undefined etiology, such as neurodegenerative conditions, and Huntington's Disease (HD) is one of them. HD is related to overexpression of Polyglutamine (PolyQ40), its aggregation, and non-solubilization, which leads to neural, behavioral, and cognitive damage. In our study, we evaluate the effect of two sublethal doses of a prallethrin-based insecticide (P-BI), in three different Caenorhabditis elegans life stages transgenerational, neonatal, and lifespan. We evaluated the Body bends and pharyngeal pumping rate, and social feeding as behavioral biomarkers. As well as acetylcholinesterase activity (AChE), PolyQ40 aggregation, antioxidant enzymes, and heat shock protein (HSP) expression. We observe that the toxic effect of P-BI is more pronounced on transgenerational and lifespan exposure. Both sublethal doses of P-BI decreased the AChE activity and retard the HSP expression as well as increased the PolyQ40 aggregates indicating a clear biomarker for possible effect in the progression of the HD, by the environmental contamination.

Persistent and transgenerational effects of pesticide residues in zebrafish.

Highly toxic chemical compounds are present in rivers and lakes, endangering the survival of non-target species. To evaluate the effects of environmental contamination on non-target species, we used the zebrafish as an animal model. Environmental concentrations of the widely used pesticides, glyphosate (GBH) at 4.8 µg·L-1 and 2,4-dichlorophenoxyacetic acid (DBH) at 3.4 µg·L-1, were used. The animals were exposed during the entire period of organogenesis and evaluated in our previous study regarding initial developmental parameters. In the present study, we evaluate these fish when achieve the adult phase, using the novel tank test (NTT) and the aversivity test. In the second step, the animals were allowed to reproduce, and the initial parameters of development, behavioral parameters in the open field test (OFT) and in the aversivity test (AST), and biochemical biomarkers as acetylcholinesterase (AChE), catalase (CAT), and superoxide dismutase (SOD) in the F1 generation were studied. Fish exposed to GBH showed hypermobility, and their anti-predatory reaction was impaired during adulthood, indicating a persistent effect. We also showed that fish had impaired behavioral and survival changes in the F1 generation as well as effects on AChE activity and antioxidant enzymes, characterizing a transgenerational effect. The fish did not show persistent effects in adulthood due to DBH exposure; however, they were unable to reproduce. Our findings demonstrate the serious impact of pesticides on fish, where the effects of contamination can affect future generations and compromise the species' survival.

Dietary transference of 17α-ethinylestradiol changes the biochemical and behavioral biomarkers in adult zebrafish (Danio rerio).

The endocrine disruptors (ED), even in low concentration, can change the homeostasis of an organism through the biochemical and physiological pathways; and are gaining more relevance due to their well-reported presence in the natural environment. EDs mainly affect non-target animals, which can bioaccumulate, leading to changes in metabolism. Another problem is due to several organisms that compose the aquatic biota serving as a basis of the food chain and transferring it to higher trophic levels. Here we evaluated the dietary transference of 17α-ethinylestradiol (EE2), in adult zebrafish chronically fed by EE2-bioaccumulated brine shrimp (BS). For this, we evaluated behavioral biomarkers such as the novel tank test (NTT), social preference test (SPT), mirror-induced aggressivity (MIA), and biochemical biomarkers such as acetylcholinesterase (AChE), superoxide dismutase (SOD), catalase (CTL), and glutathione-S-transferase (GST) activity, cortisol, and lipid peroxidation levels in adult zebrafish. The behavioral effects can be explained by the changed effects on acetylcholinesterase activity as well as in the antioxidant system mainly affected by the high levels of EE2 identified by HPLC shown that had occurred during a dietary transfer for fish. EE2 has a potential pattern for bioaccumulation and dietary transfer in biological tissue and EE2 can affect the behavior of fish. The observed effects could be dangerous to the environment, affecting, other animals and even human health.

Deferoxamine Supplementation Abolished Iron-Related Toxicity of Ilex paraguariensis Extract: Behavioral and Biochemical Evaluation in Adult Zebrafish (Danio rerio).

Ilex paraguariensis (Herb mate) is a native plant from South America, widely consumed through the infusion of dried leaves. The presence of antioxidant properties in herb mate may be relevant and contribute to evaluating the effect of its compounds against oxidative stress, which could cause neurodegenerative diseases. Despite having health benefits, there are reports of the presence of heavy metals in extracts obtained from the infusion. One of these metals is iron (Fe), found in large amounts in herb mate. To reverse the cumulative effects of metals and Fe in the body, the use of Deferoxamine (Dfx) is indicated, being a potent chelator of Fe. In this work, we aimed to evaluate the antioxidant potential of the micro-encapsulated extract of I. paraguariensis (MEIP) supplemented with Dfx on zebrafish behavior and biochemical biomarkers. To evaluate the effect per se and the supplementation, four groups were established: the first group was the control (water); the second, fish treated with MEIP; the third group was formed of fish treated with Dfx; while the fourth group was treated with both MEIP and Dfx. When applied alone, Dfx presents an anxiogenic-like pattern on zebrafish (Danio rerio), while the MEIP shows an anxiolytic-like behavior. The antioxidant enzymes are re-modulated close to control when the MEIP + Dfx is applied. The cholinergic system shows an activation of the signaling, as well as the heme radical group formation, which is not affected by the Dfx-chelating effect. Thus, the supplementation of MEIP with Dfx is important to transform this extract into one that is safer and healthier for human consumption.

Impaired initial development and behavior in zebrafish exposed to environmentally relevant concentrations of widely used pesticides.

Pesticides reach water bodies through different routes, either owing to incorrect packaging disposal, direct application to control macrophytes, leaching from fields, or natural degradation processes. In the aquatic environment, adverse effects in non-target species that come in contact with these substances are poorly understood. Currently, the most used pesticides are glyphosate (GBH) and 2,4-dichlorophenoxyacetic acid-based herbicides (DBH), as its presence in water bodies is already known, we used environmental concentrations and our exposure time comprised the entire period of organogenesis (3-120 h post-fertilization). We evaluated the response of embryos in their early development with the parameters of mortality, hatching, spontaneous movement, and heart rate; and it's through behavior the open field test and aversive stimulus, as well as biochemical analyzes of acetylcholinesterase activity (AChE), catalase (CTL) and superoxide dismutase (SOD) as a possible mechanism of action. Exposure to GBH decreased survival, caused hypermobility and anxiolytic behavior, negatively affected the anti-predatory behavior of the larvae, and increases acetylcholinesterase activity, whereas exposure to DBH caused only slight hypermobility in the larvae and increases acetylcholinesterase activity. These changes may compromise the perpetuation of the species, the search for partners/food, and facilitate the action of predators, which can result in serious ecological consequences.

Synthetic estrogen bioaccumulates and changes the behavior and biochemical biomarkers in adult zebrafish.

Estrogen is considered to be an endocrine disrupter and is becoming increasingly more prevalent in the daily life of humans. In some cases, estrogen is not fully metabolized by organisms and may be excreted in either its original form or in organic complex forms, into water residue systems reaching concentrations of 0.05 ng.L-1 to 75 ng.L-1. However, estrogen 17α-ethinylestradiol (EE2), which is used in oral contraceptives, is very difficult to remove from water. Here, we evaluated whether the synthetic hormone, EE2, affects the nervous system and the behavior of adult zebrafish. We established a range of concentrations (0.05, 0.5, 5, 50, and 75 ng.L-1), in addition to the control, to evaluate the effect of this compound and its bioaccumulation in zebrafish tissues. Here we show that EE2 bioaccumulates in fish and can change its behavior with an increased time in the upper zone (novel tank test) and far from the shoal segment (social preference test), demonstrating a clear anxiolytic pattern. The anxiolytic effect of EE2 can be harmful as it can affect the stress response of the species. The results presented herein reinforce the idea that the presence of EE2 in environmental water can be dangerous for non-target animals.

Neuroprotective and antioxidant effects of pitaya fruit on Cu-induced stress in adult zebrafish.

The harmful effects of heavy metals on organisms have not been fully described. At present, there is a close relationship between neurodegenerative diseases and copper toxicity. In addition, the copper effect on the central nervous system is followed by high levels of free radicals in different body tissues. An increase in free radical levels leads to aging-related diseases, resulting in the appearance of senile comorbidities. An increase in the consumption of natural compounds that could help to reduce this dangerous effect on organisms was observed. Pitaya (Hylocereus undatus) is a tropical fruit with great antioxidant potential and can help the organism with oxidative damage. This study evaluated the effect of H. undatus on zebrafish organisms in front of copper-induced toxicity. Therefore, the behavior, cholinergic system, antioxidant enzymes, and ALAD activity were evaluated as biomarkers. Our results highlight the great potential of Pitaya's pulp to reduce the levels of anxiety and aggressivity in fish and reduce cortisol levels. It could mediate the normal response of the cholinergic nervous system, antioxidant enzymes, and ALAD activity. Therefore, our data suggest that pitaya might improve the senile aging. PRACTICAL APPLICATIONS: The potential practical application of this study is related to the neuroprotective effect of the Hylocereus undatus microencapsulated pulp extract against metal-induced impairments. The results have shown that this extract is able to reduce the copper-induced damage modulating the antioxidant system and the cholinergic nervous system. One of the implicated potentials of use for this extract is the food supplementation to its fortification.

Pitaya fruit extract ameliorates the healthspan on copper-induced toxicity of Caenorhabditis elegans.

Copper (Cu) is an essential metal and it is important for metabolism. However, in high concentrations, it becomes toxic. Metal-induced toxicity is the cause of many neurodegenerative diseases. So it is necessary to search mechanisms to find ways of healthy aging. Natural compounds and diets based on fruits are increasingly common and could lead to a healthy life. Pitaya (Hylocereus undatus) is a tropical and Latin American, fruit that is gaining more popularity due to its antioxidant properties. Here, we evaluate the preventive and curative effect of different doses of microencapsulated pulp H. undatus extract on copper-induced toxicity. For this we use the nematode Caenorhabditis elegans, to investigate the effects of pitaya extract on behavior, lipid peroxidation, antioxidant chaperon, and cholinergic nervous system (ColNS). Results showed behavioral changes, decreased cell death biomarkers, and lipid peroxidation caused by copper, and these toxic effects were prevented and reverted by Pitaya's extract. After all, the extract can be used in diet as a supplement and studied to treat or prevent specific diseases, some of them linked to contamination and senility-related conditions. PRACTICAL APPLICATIONS: This research has been aimed to provide the uses of Hylocereus undatus microencapsulated pulp extract for the prevention and treatment of copper-induced toxicity. We have been shown that Pitaya is a good source of antioxidant compounds that can ameliorate the antioxidant system as well as the cholinergic nervous system avoiding behavior changes before and after the metal toxicity of copper. Therefore, the potential applications and common use of this extract can serve as food supplementation to prevent metal oxidative damage as well as to repair clinical cases of copper poisoning.

Fruit extract of red pitaya (Hylocereus undatus) prevents and reverses stress-induced impairments in the cholinergic and antioxidant systems of Caenorhabditis elegans.

The addition of fruit to the diet is very important, and we can use nutraceutical and functional foods for this supplement. A little-known fruit is a red pitaya (Hylocereus undatus) that has been widely reported to have a high antioxidant potential. In this study, we analyzed the in vitro and in vivo antioxidant capacity of microencapsulated pitaya extract on the behavior, antioxidant, and nervous system of the nematode Caenorhabditis elegans. The worms were treated with fruit extract before and after juglone-induced stress, to determine the protective or curative effects of pitaya. We have been evaluated cholinergic, antioxidant, and behavioral biomarkers. We have evidenced that the pulp of pitaya contains antioxidant compounds and can serve as a potential nutraceutical product. In addition, the fruit extract was effective in preventing and/or reverse the stress-induced damages, even at high levels of chemical stress at all evaluated parameters. PRACTICAL APPLICATIONS: The potential applications and uses aimed by this research are related to the supplementation of foods given the antioxidant effect. Our data suggested that the effect of the pitaya fruit microencapsulated pulp extract was effective to prevent and repair the damage caused by oxidative stress. Besides the use of this microencapsulated extract can be an auxiliary in the treatment of diseases related to oxidative damage as well as promoting senescent aging. Another important use is the application of this extract as a dietary supplement to fortify the antioxidant system.