Zebrafish Maintenance Conditions Affect Behavioral and Biochemical Biomarkers: A Possible Interfering Factor on the Research Results.

Over the years, scientific research with fish models has grown at a rapid pace, and issues such as animal welfare are becoming increasingly important in various areas of animal husbandry and experimentation. Here, we evaluated whether Danio rerio behavior is affected by long-term maintenance (75 days) in an enriched environment or a chronic stress (CS) situation. In addition, we evaluated some biochemical parameters related to redox status. We concluded that long-term maintenance of zebrafish in enriched environment might induce an anxiety-like behavior pattern when these fish are faced with an acute subsequent stressor. These anxiety results, the increased school cohesion, and the absence of oxidative damage allow us to hypothesize that the fish maintained in environmental enrichment (EE) situation is more reactive, showing a strong protective reaction to the stress. From an applicable perspective, we show that both too much stress and too little stress are not ideal for zebrafish stocks. In CS situations, fish can habituate and might not respond optimally to test conditions. In opposite, the low stress promoted by environmental enrichment also renders the fish incapable of dealing with occasional stressors optimally, because now even normal conditions appear stressful to them and may elicit fear behaviors they normally would not exhibit.

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.

A single exposure to sub-lethal concentrations of a glyphosate-based herbicide or fluoxetine-based agent on growth performance in Nile tilapia.

The aim of the present study was to determine e whether a single acute 96 hr exposure of a glyphosate-based herbicide (GBH) to Nile tilapia fingerlings affected growth performance during the first 90 days of culture. This association was considered as GBH increases serotoninergic activity that affect fish anorexically. Although these findings were based upon chronic investigations, this study was designed to examine whether a single, acute, but excessive concentration GBH might impair growth performance in fish. In parallel, fish were also exposed to fluoxetine (FLU), a drug that selectively inhibits the reuptake of serotonin in brain synapses, leading to increased serotoninergic activity. Data demonstrated a decreased growth performance in fingerlings exposed to GBH or FLU compared to unexposed fingerlings. In fact, FLU-exposed fingerlings exhibited lower average weight and length, diminished weight gain, which resulted in lower final biomass. GBH-exposed fish, despite displaying a lower mean body weight, exhibited a biomass similar to biomass on controls. These body weight differences were noted after 30-60- and 90-day growth period in clean water. In an aquaculture context, these observed changes may be considered harmful to the production or economic performance of large-scale farming as currently practiced in tilapia farming.

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.

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.

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.

Transfluthrin- and prallethrin-based insecticides elicit specific enzymatic antioxidant responses in different tissue of zebrafish.

Pyrethroids prallethrin (P-BI) and transfluthrin (T-BI) are among the most commonly used molecules with insecticide action. These molecules comprise different formulations of insecticides largely used in household, agricultural, and animal production fields. However, the increased use of these molecules has led to concerns regarding their safety in animals and humans. Oxidative stress (OS) is believed to be easily established by xenobiotic contacts, such as pyrethroids. We aimed to evaluate and understand the impact of two household insecticides and two doses applied to different tissues of the antioxidant system of zebrafish (Danio rerio). We observed that the effect on the antioxidant system differed between tissues. The muscle was the most affected tissue in the body, the antioxidant enzymes were activated, and a mechanism of non-enzymatic antioxidants was activated; however, it could still cause cellular damage. The observed effect on muscle may be related to the progression of neurodegenerative conditions. In addition, in the brain, these compounds can inactivate the first line of enzymatic antioxidant defense, which is compensated for by the second line, avoiding cellular damage. Ultimately, the gill tissue did not appear to suffer lipid damage, but heme group formation was largely affected by the compounds.

Photobiomodulation Reduces the Cytokine Storm Syndrome Associated with COVID-19 in the Zebrafish Model.

Although the exact mechanism of the pathogenesis of coronavirus SARS-CoV-2 (COVID-19) is not fully understood, oxidative stress and the release of pro-inflammatory cytokines have been highlighted as playing a vital role in the pathogenesis of the disease. In this sense, alternative treatments are needed to reduce the level of inflammation caused by COVID-19. Therefore, this study aimed to investigate the potential effect of red photobiomodulation (PBM) as an attractive therapy to downregulate the cytokine storm caused by COVID-19 in a zebrafish model. RT-qPCR analyses and protein-protein interaction prediction among SARS-CoV-2 and Danio rerio proteins showed that recombinant Spike protein (rSpike) was responsible for generating systemic inflammatory processes with significantly increased levels of pro-inflammatory (il1b, il6, tnfa, and nfkbiab), oxidative stress (romo1) and energy metabolism (slc2a1a and coa1) mRNA markers, with a pattern similar to those observed in COVID-19 cases in humans. On the other hand, PBM treatment was able to decrease the mRNA levels of these pro-inflammatory and oxidative stress markers compared with rSpike in various tissues, promoting an anti-inflammatory response. Conversely, PBM promotes cellular and tissue repair of injured tissues and significantly increases the survival rate of rSpike-inoculated individuals. Additionally, metabolomics analysis showed that the most-impacted metabolic pathways between PBM and the rSpike treated groups were related to steroid metabolism, immune system, and lipid metabolism. Together, our findings suggest that the inflammatory process is an incisive feature of COVID-19 and red PBM can be used as a novel therapeutic agent for COVID-19 by regulating the inflammatory response. Nevertheless, the need for more clinical trials remains, and there is a significant gap to overcome before clinical trials can commence.

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.

Plasma proteome responses in zebrafish following λ-carrageenan-Induced inflammation are mediated by PMN leukocytes and correlate highly with their human counterparts.

Regulation of inflammation is a critical process for maintaining physiological homeostasis. The λ-carrageenan (λ-CGN) is a mucopolysaccharide extracted from the cell wall of red algae (Chondrus crispus) capable of inducing acute intestinal inflammation, which is translated into the production of acute phase reactants secreted into the blood circulation. However, the associated mechanisms in vertebrates are not well understood. Here, we investigated the crucial factors behind the inflammatory milieu of λ-CGN-mediated inflammation administered at 0, 1.75, and 3.5% (v/w) by i.p. injection into the peritoneal cavity of adult zebrafish (ZF) (Danio rerio). We found that polymorphonuclear leukocytes (neutrophils) and lymphocytes infiltrating the ZF peritoneal cavity had short-term persistence. Nevertheless, they generate a strong pattern of inflammation that affects systemically and is enough to produce edema in the cavity. Consistent with these findings, cell infiltration, which causes notable tissue changes, resulted in the overexpression of several acute inflammatory markers at the protein level. Using reversed-phase high-performance liquid chromatography followed by a hybrid linear ion-trap mass spectrometry shotgun proteomic approach, we identified 2938 plasma proteins among the animals injected with PBS and 3.5% λ-CGN. First, the bioinformatic analysis revealed the composition of the plasma proteome. Interestingly, 72 commonly expressed proteins were recorded among the treated and control groups, but, surprisingly, 2830 novel proteins were differentially expressed exclusively in the λ-CGN-induced group. Furthermore, from the commonly expressed proteins, compared to the control group 62 proteins got a significant (p < 0.05) upregulation in the λ-CGN-treated group, while the remaining ten proteins were downregulated. Next, we obtained the major protein-protein interaction networks between hub protein clusters in the blood plasma of the λ-CGN induced group. Moreover, to understand the molecular underpinnings of these effects based on the unveiled protein sets, we performed a bioinformatic structural similarity analysis and generated overlapping 3D reconstructions between ZF and humans during acute inflammation. Biological pathway analysis pointed to the activation and abundance of diverse classical immune and acute phase reactants, several catalytic enzymes, and varied proteins supporting the immune response. Together, this information can be used for testing and finding novel pharmacological targets to treat human intestinal inflammatory diseases.

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.

Sound stimulus effects on dusky damselfish behavior and cognition.

Anthropogenic noises are widespread and affect marine wildlife. Despite the growing knowledge on noise pollution in the marine environment, its effects on fish cognition are scarce. Here, we investigated the effects of sound exposure on anxiety-like behavior and memory retention on dusky damselfish Stegastes fuscus. The animals were trained in a conditioned place aversion task, and exposed to two daily sessions of music at intensities of 60-70 dBA or 90-100 dBA, while the control group was kept at 42-46 dBA (no music) for five days. After that, fish were tested in the novel tank paradigm and tested for the memory of the aversive task. In the novel tank, animals exposed to sound spent more time still and decreased the distance from the bottom of the tank. Animals also spent more time on the aversive side of the conditioning tank. These results suggest that anthropogenic noise applied through high-intensity music can increase anxiety and decrease memory retention in S. fuscus, suggesting the deleterious potential of noise for reef species.

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.

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.