Erythrocyte Recovery in Oreochromis niloticus Fish Exposed to Urban Effluents.

Urban activities pollute aquatic ecosystems, and the integrity of organisms such as fish. The use of cytological techniques, such as the analysis of blood cellular integrity using the Micronucleus test, can help detect mutagenic damage as a result to urban effluents exposure. In this context, this study aimed to evaluate the frequency of micronucleus and other nuclear abnormalities in Oreochromis niloticus fish environmentally exposed to urban effluents in relation to their erythrocyte recovery capacity when exposed to clean water (30 and 45 days). The results indicated high copper, dissolved iron, nickel, and thermotolerant coliform levels in the urban stream. There was no difference in the frequency of micronuclei. In contrast, cells with nuclear nuclei, binucleates, kidney-shaped nuclei, notched nuclei, lobed nuclei, and segmented nuclei decreased according to the time the fish were exposed to clean water. When exposed to clean water, we conclude that urban fish recover from genotoxic and cytotoxic damage.

Genotoxic, mutagenic, and cytotoxic analysis in bats in mining area.

Pollution generated by the mining industry can cause harm to wildlife. This study aimed to evaluate the cytotoxicity, genotoxicity and mutagenicity in bats environmentally exposed to open pit mining. Thus, 62 bats of the following species, Carollia perspicillata, Glossophaga soricina, Phyllostomus hastatus, and Desmodus rotundus exposed to mining activities (ferronickel) were used in the analysis. The animals were obtained in samplings in July and November of 2021, totaling 8 days of sampling in the field. The results indicated that species differ in the frequency of genotoxic damage between sampling points within the mining landscape. Cytotoxicity was observed by scoring of karyorrhexis, pyknosis and karyolysis. The most captured species, C. perspicillata, showed differences in DNA damage between exposed and unexposed populations, but no differences were observed between males (n = 14) and females (n = 20). G. soricina was also a sensitive species for indicating a high frequency of DNA damages compared to the omnivore P. hastatus. Elements such as Mn, Cr, Pb, and Zn observed in water samples were at high levels in the mining area. We conclude that bats in mining areas are susceptible to increased DNA damage as already identified for other species.

Micronucleus and different nuclear abnormalities in wild birds in the Cerrado, Brazil.

Free-living birds play an important role as bioindicators in natural environments. In this study, we used the micronucleus test and nuclear erythrocyte abnormalities to investigate the difference in the genotoxic damage frequency between animals in agricultural areas (soybean planting) in relation to the conserved area. It was also discussed the bird's eating habits, which are important for ecotoxicological analysis. The results point to a difference between the insectivorous, granivorous, and omnivorous guilds. The omnivore Gnorimopsar chopi was the species that most exhibited micronucleus frequency in the protected area. In the agricultural area, the animals did not differ in the genotoxic damage frequency. In the comparison between common species in both environments, G. chopi from the agricultural area showed a micronuclei frequency almost three times higher in relation to specimens collected in the conserved area. Based on these results, this study adds to the efforts of using the micronucleus test as a simple and accessible tool for biomonitoring the wild fauna. It can be concluded that the passerine, G. chopi, due to its higher genotoxic damage frequency may be a strong candidate to indicate environmental health.

Evaluation of genotoxicity in bat species found on agricultural landscapes of the Cerrado savanna, central Brazil.

Habitat loss and fragmentation together represent the most significant threat to the world's biodiversity. In order to guarantee the survival of this diversity, the monitoring of bioindicators can provide important insights into the health of a natural environment. In this context, we used the comet assay and micronucleus test to evaluate the genotoxic susceptibility of 126 bats of eight species captured in soybean and sugarcane plantation areas, together with a control area (conservation unit) in the Cerrado savanna of central Brazil. No significant differences were found between the specimens captured in the sugarcane and control areas in the frequency of micronuclei and DNA damage (comet assay). However, the omnivore Phyllostomus hastatus had a higher frequency of nuclear abnormalities than the frugivore Carollia perspicillata in the sugarcane area. Insectivorous and frugivorous bats presented a higher frequency of genotoxic damage than the nectarivores in the soybean area. In general, DNA damage and micronuclei were significantly more frequent in agricultural environments than in the control area. While agricultural development is an economic necessity in developing countries, the impacts on the natural landscape may result in genotoxic damage to the local fauna, such as bats. Over the medium to long term, then DNA damage may have an increasingly negative impact on the wellbeing of the local species.

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.

Multi-taxon inventory and landscape characterization in an agrosystem of the Brazilian Midwest targeted for payment for environmental services / Inventário multi-táxon e caracterização da paisagem em um agrossistema no Centro-Oeste brasileiro alvo de pagamento por serviços ambientais

Abstract: The replacement of natural landscapes by anthropic environments has led to habitat loss and consequently, to the decline and extinction of many species populations. Several strategies, such as the creation of protected areas (PAs) and payment for ecosystem services have been adopted to mitigate the consequences of this biodiversity crisis. In Rio Verde, Goiás, Brazil, a Water Producer Program (WPP) was established, which provides payment for ecosystem services for the maintenance of riparian vegetation in the region of the Ribeirão Abóbora, a river which supplies the city and metropolitan area. In the absence of environmental diagnostic studies in the region, this present study contributes to reducing some knowledge gaps, through the: (i) characterization of the landscape in the WPP area, (ii) characterization of species of fauna (amphibians, birds, fish, and medium and large terrestrial mammals) and woody flora that occur in this area, and (iii) comparison of species diversity (fauna and flora) found in our study area with the diversity observed in the Cerrado protected area network. Our results reveal a low native vegetation cover (~21% of total area), but we recorded 300 total species: 20 species of anuran amphibians, 100 birds, 10 fish, 16 terrestrial mammals (medium and large), and 154 woody plants. Our sample comprises species considered endemic to the Cerrado (e.g., Anuran - Barycholos terntezi and Mammal - Callithrix penicillata), threatened species (e.g., Mammals - Herpailurus yagouaroundi, Myrmecophaga tridactyla and Tapirus terrestris), and exotic invasive species (Mammal - Sus scrofa). The results presented in this study are relevant and may help in the proposal of management actions within the scope of this important program of payment for environmental services.

Resumo: A substituição das paisagens naturais por ambientes antropizados tem levado à perda de habitat e, consequentemente, ao declínio populacional e extinção de muitas espécies. Várias estratégias como a criação de Unidades de Conservação (UCs) e o pagamento de serviços ecossistêmicos têm sido adotadas para mitigar as consequências da crise de biodiversidade. Em Rio Verde, estado de Goiás, foi estabelecido um Programa Produtores de Água (PPA) que prevê o pagamento de serviços ecossistêmicos pela manutenção da vegetação ripária na região do ribeirão Abobora que abastece a cidade. Diante da ausência de estudos de diagnóstico ambiental da região, o presente estudo contribui para reduzir algumas lacunas de conhecimento, através da: i) caracterização da paisagem na área deste PPA, ii) caracterização das espécies da fauna (anfíbios, aves, peixes e mamíferos terrestres de médio e grande porte) e flora lenhosa que ocorrem neste local, iii) comparação da diversidade de espécies (fauna e flora) encontrada na área de estudo com aquela observada na rede de áreas protegidas do Cerrado. Nossos resultados revelam baixo índice de cobertura vegetal nativa (~21% da área total), mas por outro lado, registramos 300 espécies, sendo 20 de anfíbios anuros, 100 aves, 10 peixes, 16 mamíferos terrestres (médio e grande porte) e 154 plantas lenhosas. A nossa amostragem compreende espécies que são consideradas endêmicas do Cerrado (p.ex. Anfíbio - Barycholos terntezi e Mamífero - Callithrix penicillata), ameaçadas de extinção (p.ex. Mamíferos - Herpailurus yagouaroundi, Myrmecophaga tridactyla e Tapirus terrestris) ou exótica invasora (Mamífero - Sus scrofa). Os resultados apresentados no presente estudo são relevantes e podem auxiliar na proposição de ações de manejo no âmbito deste importante programa de pagamento por serviços ambientais.

Current Status of Ecotoxicological Studies of Bats in Brazil.

Bats are sensitive to contaminants generated by agricultural activities, mining, and urbanization. In this review, we update the status of bat toxicology in Brazil. Agriculture, for example, in addition to habitat fragmentation and loss, undoubtedly affects non-target organisms through the use of pesticides. Other factors such as trace metals are a neglected problem in the country, as they can deposit on insects and plants reaching bats through the ingestion of these foods. Of the 184 species of bats in the country, only 4.9% have been investigated. The frugivorous species, Artibeus lituratus, has frequently been studied for the effects of pesticide exposure, and impacts at the cellular level on metabolism and reproduction have been observed. Given the scarcity of studies on bat ecotoxicology, we encourage national researchers and scientists elsewhere to increase knowledge of the effects of chemical contaminants on bats in Brazil.

Micronucleus test in tadpole erythrocytes: Trends in studies and new paths.

The micronucleus test has been applied for more than three decades in tadpoles, generating an early warning of environmental quality. In this study, we reviewed 48 articles on the micronucleus test in tadpoles, published between 1987 and 2018. The findings reveal that pesticides have been the main topic discussed in the induction of micronucleus and other nuclear abnormalities in anuran larvae to the detriment of the widespread use of compounds used in agriculture. In addition to pesticides, a number of other xenobiotic agents have been targeted for genotoxic damage, such as heavy metals, radiation and wastewater. An appeal is reported to environmental contaminants, which when released naturally into the environment or because of human activities may contaminate aquatic habitats, threatening populations of tadpoles that depend on these environments for their survival. Larvae can bioaccumulate these contaminants that cause progressive impacts, ranging from DNA damage to metamorphosis delays, as well as malformations. We found that Argentina is the main driving force for the application of this test in anuran larvae along with Brazil. Different erythrocyte malformations have been reported for the erythrocyte nuclear abnormalities test, binucleated cells, nuclear buds, notched, lobed, reniform, nuclear bebbled, anucleated, picnotic and apoptotic cells are the most cited. In summary, the presence of chemical or physical agents, along with other disturbances of the habitat, can have a significant impact on the life history of the species, contributing to the decline of anuran populations.

Genotoxic Evaluation in Tadpoles Associated with Agriculture in the Central Cerrado, Brazil.

Many agricultural practices cause environmental degradation that affects the cellular integrity of anurans. In the present study, we provided in situ data of Dendropsophus minutus, Physalaemus cuvieri, and Scinax fuscovarius collected in soybean/corn and conservation units in the Brazilian Cerrado. The in situ data showed no significant variation in the micronucleus frequency between the sites, only the reniform cells had a higher rate for the agricultural environment. A combined analysis of all nuclear erythrocyte abnormalities (ENAs = nuclear buds, reniform nuclei, apoptotic cell, binucleated, and anucleated cells) was recorded higher frequencies in farmland. Overall, Scinax fuscovarius was considered the best potential bioindicator for soybean/corn plantations. Finally, we recommend expanding the micronucleus test for in situ studies to expand our understanding of the sensitivity of native anuran species and provide a more systematic assessment of the adverse effects of environmental pollutants on wildlife.

Micronucleus and other nuclear abnormalities in exfoliated cells of buccal mucosa of bats at different trophic levels.

The micronucleus (MN) test in exfoliated cells of the buccal mucosa is a relatively non-invasive method for the monitoring of populations exposed to genotoxic risks. In this study, the MN test was used as bats conservation strategy. The highest frequencies of micronuclei were recorded in the frugivorous bats sampled in both urban and agricultural environments, as well as in insectivorous bats from the urban zone. Female of this group (insectivorous) presented higher frequency of MN when compared to males. Other guilds showed no difference in gender assessments in each environment, as well as in the correlation between weight and MN. In addition to micronuclei, a number of other types of nuclear abnormality were recorded, including binucleated cells and karyolysis in the frugivores from the agricultural environment. Binucleated cells were also relatively common in urban frugivores and insectivores, and karyolysis was common in insectivores. Nectarivorous bats did not exhibit a significant increase in any type of nuclear abnormality in either environment. In summary, study results indicate that buccal mucosa of bats is a sensitive site for detecting micronuclei and other nuclear abnormalities. However, more research is needed to indicate whether xenobiotic agents are affecting this cellular integrity.

Genotoxicological analyses of insectivorous bats (Mammalia: Chiroptera) in central Brazil: The oral epithelium as an indicator of environmental quality.

The micronucleus (MN) test of the human buccal mucosa was developed more than 30 years ago, although this technique has only recently been applied to wild mammals. This paper presents a pioneering study in the genotoxicological evaluation of the exfoliated cells of the buccal mucosa of bats. The assay was applied to two insectivorous bat species (Noctilio albiventris and Pteronotus parnellii) sampled in riparian corridors located in the city of Palmas (capital of the Brazilian state of Tocantins), with the results being compared with those obtained for a third insectivorous species (Nyctinomops laticaudatus), which has established a colony under a road bridge in the same region. This colony represents one of the largest molossidae populations ever recorded in Brazil. A significantly higher frequency of micronuclei was recorded in this colony, as well as a number of other nuclear abnormalities, including binucleated cells, cells with condensed chromatin and karyolysis, in comparison with the bats from the riparian corridors, indicating that the bats from the bridge colony are more susceptible to genotoxic damage. Thus, it is demonstrated the importance of the biomarker (MN) for use in wild animals and allows to conclude that colony bats are more susceptible to genotoxic damages.

Monitoring the morphological integrity of neotropical anurans.

Amphibians are considered to be excellent bioindicators to their morphophysiological characteristics and life cycle. In this context, the present study investigated the morphological integrity of anuran larvae collected in preserved environments in the Emas National Park, in the municipality of Mineiros (Goiás state, Brazil), and in environments representative of the agricultural matrix of the Rio Verde region, also in Goiás, where there is a long history of the use of agricultural pesticides. Samples of water from temporary ponds, permanent dams, and veredas were analyzed for the presence of pesticides and, especially atrazine (5350 µg/L), found at significantly higher concentrations in the agricultural matrix. We observed a high percentage (approximately 10%) of morphological malformations including alterations of the fins in Boana albopunctatus and Scinax fuscovarius; alteration in oral structures in B. albopunctatus, Dematonotus muelleri, Physalaemus centralis, Physalaemus cuvieri, and Leptodactylus fuscus mainly in the tadpoles collected in the agricultural environment in comparison with those from the protected area (3.5%; P < 0.0001, χ2 31.75). However, changes in the eyes, mouth, intestines, and nostrils, as well as amelia were observed only in the agricultural environment. The vast majority of the observed malformations were associated with the tail and oral disc, which suggests that these anatomical parameters may be used as sensitive morphological biomarkers. Given these findings, we reinforce that areas of agricultural land may have a deleterious effect on the morphological integrity of the tadpoles and consequently, on their development, and that these features may be used as indicators of environmental quality and health.

The micronucleus test for the oral mucosa: global trends and new questions.

This study reviews global trends in the publication of papers on the micronucleus test of the exfoliated cells of the oral mucosa in mammals as an approach for environmental biomonitoring. This test has been widely used due to its precision for the detection of chromosome damage. Our temporal analysis showed a significant increase (p < 0.05) in the publication of papers on the oral mucosa over the past 33 years. Brazil was the country that published most papers (24% of the total), followed by India (16%), the USA (10%), Mexico (7%), and Turkey (6%). A further 30 countries contributed the other 37% of the papers. Overall, 99% of the micronucleus studies involved human subjects, and only 1% involved other mammals. As many wild mammals are subject to the same environmental pressures as humans, in particular chemical pollution, it seems likely that many species are equally susceptible to genotoxicogical damage. We emphasize the importance of applying this technique to the analysis of the oral mucosa of wild mammals, as well as the evaluation of its effectiveness, as observed in humans, and the expansion of the available approaches to the monitoring of environmental quality.

Effects of thermal stress on hepatic melanomacrophages of Eupemphix nattereri (Anura).

Melanomacrophages are the pigmented cells present in the hematopoietic organs. Besides melanin, hemosiderin and lipofuscin are also observed in the melanomacrophages. For the liver, however, numerous studies relate these cells to immunological and metabolic functions. Therefore, the aim of this study is to evaluate the hepatic metabolism by quantifying melanin, hemosiderin and lipofuscin in the anuran Eupemphix nattereri submitted to varying thermal conditions. E. nattereri adult males were separated into three groups, as follows: (i) five animals in the control group were kept at room temperature (27°C); (ii) 30 animals were submitted to hyperthermic (35.1°C); and (iii) 30 to hypothermic (18.9°) conditions. In each experiment, the animals were analyzed and separated into two different treatments: (1) immediately after undergoing the stress; and, (2) after recovering from the stress caused by the stimulus, at three distinct times (12 hr, 24 hr, and 48 hr). Both hyperthermia and hypothermia decreased hepatic pigmentation after thermal stress. The recovered animals of both experimental treatments showed as much pigmentation as the control animals. Thermal stress alters hemosiderin and lipofuscin as well, which may be related to liver function catabolism. In conclusion, liver pigmentation decreased due to temperature variation and duration of thermal stimulation to which the animals were exposed. The increase in temperature rather than hypothermia led to more drastic physiological disorders. In this study, we observed that thermal stress for a short period compromises the morphology and liver function, as observed by the changing pigmentation of melanomacrophages. These analyses can be used as biomarkers of environmental effects.