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.

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.

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.

Effect of multiple stressors and population decline of frogs.

The ongoing decline in anuran populations is linked primarily to the effects of stressor agents such as pathogens, pesticides, alterations of natural landscapes, and the introduction of exotic species. Most studies that have evaluated the effects of these stressors have focused on a single component, which is the opposite of the reality of most natural environments, where anuran populations tend to suffer the influence of multiple agents simultaneously. Studies of the effects of the interaction between these components are extremely important, given that one agent may potentialize (synergistic effect) or weaken another (antagonistic effect) or, in some cases, have a neutral effect. The present study is based on the scientometric analysis of three bibliographic databases (ISI Web of Science, Scopus, and PubMed), which identified 1376 papers that reported on the global decline of anuran populations, although only 172 of these studies focused on the interactive effects of environmental stressors. Synergistic effects were the most frequent type of interaction, followed by antagonistic effects, and a small number of studies that found no clear interaction between the stressors. Pathogens and pesticides were the classes of stressor studied most frequently, while climate-pathogen and pathogen-pesticide interactions were the combinations that featured in the largest number of studies. Overall, we would recommend a more systematic focus on the dynamics of the interactions among the stressors that impact anuran populations, in particular for the elaboration of conservation programs, given that these agents tend to have complex combined effects.

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.

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.

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.

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.

Visceral pigmentation in three species of the genus Scinax (Anura: Hylidae): distinct morphological pattern.

Amphibians share with other ectothermic vertebrates an extracutaneous pigmentary system consisting of melanin-containing cells in various organs and tissues. We describe the interspecific variation in the visceral pigmentation in three anuran species (Scinax similis, S. fuscovarius, and S. fuscomarginatus). We analyzed the visceral pigmentation of 15 adult males from each species during the reproductive period. The individuals were weighed and measured, and the pigmented visceral cells were classified and documented in stereomicroscope. The shape and amount of pigment cells differed among organs and also among species. Significant differences were detected in the cardiorespiratory system, digestory system, urogenital system, and lumbosacral peritoneum. In the urogenital system, the main difference was observed in the testes, in which only S. fuscomarginatus had varying degrees of pigmentation.

Morphological changes of the epididymis and description of the excurrent ducts of Phrynops geoffroanus (Testudines: Chelidae) during the reproductive cycle.

The seminal ducts (efferent ductule, epididymis, and deferent duct) in adults of Phrynops geoffroanus were examined using light microscopy. A series of tubules (efferent ductules) connect the testes to the epididymides. The efferent ductules are formed by a rete of small tubules of varying diameters, with simple columnar epithelium formed by the ciliated cells, nonciliated cells, and few basal cells. The epididymis is a simple, long and highly convoluted tubule that receives the efferent ductules throughout its extension. It is covered by a pseudostratified columnar epithelium with three cellular types: the principal cells, which are the most abundant, basal cells, and a small narrow cell. The histological differences in the epididymis region (cranial, medial, and caudal), as well as the differences in the epithelium throughout the reproductive cycle, are discussed. The deferent ducts consist of a low pseudostratified epithelium with two cellular types: the principal and basal cells. During the months analyzed, spermatozoa were stored in the epididymis, and deferent ducts were found.

Pigmentation in anuran testes: anatomical pattern and variation.

In amphibians, pigmented cells are present in several organs, composing an extracutaneous pigmentary system. Seventeen species from two families were studied to develop a protocol for pigmentary classification. The amount and distribution of these cells are variable, allowing the establishment of anatomical patterns for visceral pigmentation in anuran testes.

Histological aspects and structural characteristics of the testes of Dendropsophus minutus (Anura, Hylidae).

The present study describes morphological aspects of testes and presents a general characterization of the seminiferous elements of Dendropsophus minutus (Peters, 1872). Twenty samples of the species were used; after macroscopic descriptions the testes were submitted to histological routine for microscopic analysis. Anatomically, the testes measured 1.90+/-0.13 mm, and were oval and milky-white. In relation to microscopic aspects, it was observed in D. minutus, as well as in anuran amphibians, that spermatogenesis occurs in the seminiferous tubule where elements of the germ epithelium are organized in spermatogenetic cysts. Each cyst contains cells in the same stage of differentiation. Characterization of each cellular type enables the identification and differentiation of germ lineage cells. Spermatogonia I, found at the epithelial base, are the largest of the lineage cells and are usually present in association with Sertoli cells present next to the basal membrane. During the mitotic proliferation phase, cysts containing variable numbers of spermatocytes II are originated; these spermatocytes are smaller and similar inside the cyst. Spermatocytes I are developed after some morphological changes; these spermatocytes are large cells with a spherical nucleus and different degrees of nuclear compaction. Spermatocytes II, highly numerous cells resultant from the first meiotic division, are much smaller than their antecedents. A second meiotic division produces haploid cell formation, specifically spermatids I, which through cellular differentiation form spermatids II. Spermatids II are elongated and organized in bundles supported by Sertoli cells. Spermatozoids appear during the spermiogenesis process and in their most advanced stage they lose their fascicular organization and are released in the tubular lumen, where they follow a pathway through the duct system.