Genotoxic effect of two environmentally safe doses of cadmium on the hepato-nephrocytic system of Bombus atratus forager workers.

Eusocial bees are declining due to anthropogenic actions. Individuals can be exposed to contaminants like Cd, which have been found in pollen. Thus, we evaluated the potential genotoxicity of Cd to the hepato-nephrocitic system of Bombus atratus foraging workers exposed to environmentally safe doses of Cd (0.001 mg/kg and 0.003 mg/kg) during 72 h. To assess the genotoxicity of Cd, we conducted an alkaline comet assay. Doses of 0.001 mg/kg of Cd caused comets of levels 2 and 3, and 0.003 mg/kg of Cd induced comets of level 4, while controls exhibited regular nucleoids. Also, 0.003 mg/kg doses caused higher Damage Index, Damage Frequency, and Total Damage. Our results bring new evidence that 0.003 mg/kg Cd exerted genotoxicity to the HNS cells of B. atratus, probably affecting secondary metabolism and the detoxification capability of bees.

Pointing out geographic and gender disparities related to productivity indicators among Brazilian ecotoxicologists.

Policies and actions related to diversity, equity, and inclusion have been discussed recently in Brazil, but there is still limited information available for most academic and knowledge fields, including ecotoxicology. This study aimed to describe the profile of Brazilian ecotoxicologists and assess gender and geographical disparities, especially regarding productivity indicators. An ecological study was conducted using data of the researchers registered in the open-access database of the Brazilian Society of Ecotoxicology, along with their respective curriculum data available on the Lattes Platform, which is the main registry portal for Brazilian researchers. The target population of our study was ecotoxicology researchers in academic educational institutions with a focus on human resource training. The data, collected in 2021, led to the inclusion of a total of 177 researchers in this study, with 62 men and 115 women, and 42.37% working in the southeastern region. Female researchers generally showed lower rates of scientific productivity when the analyzed metrics included research productivity scholarships, the number of articles published in scientific journals, and the number of articles published with international collaborations, compared to male researchers. Researchers from the southeastern and southern regions also had higher rates of productivity and human resource training compared to those from other regions of the country. The proportion of funding grants received was significantly higher among men, and this profile was also observed among researchers from the southern and southeastern regions, although with a smaller disparity. This means that there is an uneven distribution of funding grants from funding organizations, which favors men and urban centers. Our findings highlight gender and geographic disparities in the scientific production of ecotoxicologists working in Brazil and reinforce the existence of complex obstacles that need to be addressed and combated within scientific societies. Integr Environ Assess Manag 2024;00:1-8. © 2024 SETAC.

Estrogen contamination increases vulnerability of amphibians to the deadly chytrid fungus.

Aquatic contaminants and infectious diseases are among the major drivers of global amphibian declines. However, the interaction of these factors is poorly explored and could better explain the amphibian crisis. We exposed males and females of the Brazilian Cururu Toad, Rhinella icterica, to an environmentally relevant concentration of the estrogen 17-alpha-ethinylestradiol (an emerging contaminant) and to the chytrid infection (Batrachochytrium dendrobatidis), in their combined and isolated forms, and the ecotoxicity was determined by multiple biomarkers: cutaneous, hematological, cardiac, hepatic, and gonadal analysis. Our results showed that Cururu toads had many physiological alterations in response to the chytrid infection, including the appearance of cutaneous Langerhans's cells, increased blood leukocytes, increased heart contraction force and tachycardia, increased hepatic melanomacrophage cells, which in turn led to gonadal atrophy. The estrogen, in turn, increased the susceptibility of the toads to the chytrid infection (higher Bd loads) and maximized the deleterious effects of the pathogen: reducing leukocytes, decreasing the contraction force, and causing greater tachycardia, increasing hepatic melanomacrophage cells, and leading to greater gonadal atrophy, which were more extreme in females. The exposure to estrogen also revealed important toxicodynamic pathways of this toxicant, as shown by the immunosuppression of exposed animals, and the induction of the first stages of feminization in males, which corroborates that the synthetic estrogen acts as an endocrine disruptor. Such an intricate relationship is unprecedented and reinforces the importance of studying the serious consequences that multiple environmental stressors can cause to aquatic populations.

The amphibian's spleen as a source of biomarkers for ecotoxicity assessment: Historical review and trends.

Amphibians are very sensitive to many environmental changes, so these animals are considered good bioindicator models for ecotoxicology. Given the importance of the amphibian spleen for hematopoietic and immune responses, this can be a key organ for the evaluation of biomarkers to monitor the health of individuals in nature or in captivity. In this systematic review, we searched databases and summarized the main findings concerning the amphibian spleen as a source of possible biomarkers applied in different scientific fields. The searches resulted in 83 articles published from 1923 to 2022, which applied the use of splenic samples to evaluate the effects of distinct stressors on amphibians. Articles were distributed in more than twenty countries, with USA, Europe, and Brazil, standing out among them. Publications focused mainly on anatomical and histomorphological characterization of the spleen, its physiology, and development. Recently, the use of splenic biomarkers in pathology and ecotoxicology began to grow but many gaps still need to be addressed in herpetological research. About 85 % of the splenic biomarkers showed responses to various stressors, which indicates that the spleen can provide numerous biomarkers to be used in many study fields. The limited amount of information on morphological description and splenic anatomy in amphibians may be a contributing factor to the underestimated use of splenic biomarkers in herpetological research around the world. We hope that this unprecedented review can instigate researchers to refine herpetological experimentation, using the spleen as a versatile and alternative source for biomarkers in ecotoxicology.

Differential liver histopathological responses to amphibian chytrid infection.

Amphibians have been facing a pandemic caused by the deadly fungus Batrachochytrium dendrobatidis (Bd). Although studies have elucidated cutaneous and homeostatic disturbances, it is still unknown if the hepatic function can be affected or if hepatic effects differ among host species. Thus, we evaluated the effects of an experimental Bd infection on the liver (histopathology and the hepatosomatic index) of 2 anuran species (Xenopus laevis and Physalaemus albonotatus) with different susceptibilities to Bd infection and compared them to uninfected controls. Bd infection increased the melanomacrophage cell area and induced leukocyte infiltration in both species. The effects were more pronounced in the sensitive species, P. albonotatus, which showed severe reduction in glycogen stores and liver atrophy, due to energetic imbalance. Hepatocytes of P. albonotatus also showed ballooning degeneration (vacuolization), which could lead to cell death and liver failure. Our results provide evidence that although the sensitive species showed more severe effects, the tolerant species also had hepatic responses to the infection. These findings indicate that hepatic function can play an important role in detoxification and in immune responses to chytridiomycosis, and that it may be used as a new biomarker of health status in chytrid infections.

Novel findings on the impact of chytridiomycosis on the cardiac function of anurans: sensitive vs. tolerant species.

BACKGROUND: Understanding of the physiological effects of chytridiomycosis is crucial to worldwide amphibian conservation. Therefore, we analyzed the cardiac function of two anuran species (Xenopus laevis and Physalaemus albonotatus) with different susceptibilities to infection by the causative agent of chytridiomycosis, Batrachochytrium dendrobatidis (hereafter Bd). METHODS: We analyzed the in situ heart rate (f H - bpm), relative ventricular mass (RVM -%), and Ca2+ handling in heart of Bd infected animals compared to uninfected controls of both study species. RESULTS: Bd infection resulted in a 78% decrease in contraction force values in P. albonotatus when compared to the less susceptible X. laevis. This negative effect was even more evident (82%) for the cardiac pumping capacity. The time to reach peak tension was 125% longer in P. albonotatus than in X. laevis, and cardiac relaxation was 57% longer. DISCUSSION: These results indicate a delay in the cardiac cycle of P. albonotatus on a beat-to-beat basis, which was corroborated by the bradycardia observed in situ. In summary, Bd-sensitive species present impaired cardiac function, which could be a factor in mortality risk. The more pronounced effects of Bd in P. albonotatus may not only result from electrolyte imbalance, as previously reported, but also could be an effect of toxins produced by Bd. For X. laevis, the ability to promote cardiac adjustments seems to be an important homeostatic feature that allows greater tolerance to chytridiomycosis. This study provides new physiological mechanisms underlying the tolerance or susceptibility of amphibian species to chytridiomycosis, which determine their adaptability to survive in the affected environments.

Impact of an environmental relevant concentration of 17α-ethinylestradiol on the cardiac function of bullfrog tadpoles.

This study evaluated if a concentration of 17α-ethinylestradiol (EE2 - 10 ng L(-1) for 96 h) normally found in Brazilian surface waters exerts any impact on cardiac function of bullfrog tadpoles (25 Gosner stage), Lithobates catesbeianus. During exposure, the animals' activity level (AL -% of active individuals) was monitored twice a day. Then, the in loco heart rate (f(H) - bpm) was determined, as well as the relative ventricular mass (RVM - % of body mass). Afterwards, cardiac ventricles were mounted for isometric force recordings (CS - mN mm(-2)), and determination of the cardiac pumping capacity (CPC - mN mm(-2) min(-1)). EE2 did not affect tadpoles' AL, although it resulted in a tachycardia in animals exposed to EE2 (f(H) = 66 bpm) when compared to controls (f(H) = 52 bpm), suggesting that EE2 acts directly on the cardiac muscle of tadpoles, rather than being a result of an increased cardiac demand due to a higher activity level (i.e., avoidance response). Additionally, EE2 exerted a positive inotropic response, which resulted in a higher CPC, which occurred independently of an increase in the number of myofibrils of EE2-exposed animals, since RVM remained similar between experimental groups. Thus, the increase on cardiac demand induced by the exposure to EE2 elevates considerably the animal energy expenditure, diverting a large amount of energy that tadpoles could use for their growth and development. These alterations can make amphibians more susceptible to predators and reduce the likelihood to reach reproductive stage.