Assuntos

RESUMO

Although non-front fanged snakes account for almost two-thirds of snake diversity, most studies on venom composition and evolution focus exclusively on front-fanged species, which comprise most of the clinically relevant accidents. Comprehensive reports on venom composition of non-front fanged snakes are still scarce for several groups. In this study, we address such shortage of knowledge by providing new insights about the venom composition among species of Phalotris, a poorly studied Neotropical dipsadid genus. Phalotris are known for their specialized venom delivery system and toxic venoms, which can cause life-threatening accidents in humans. We evaluate the venom-gland transcriptome of Phalotris, comparing the following three South American species: P. reticulatus for the Araucaria Pine forests, P. lemniscatus for the Pampa grasslands, and P. mertensi for the Brazilian Cerrado. Our results indicate similar venom profiles, in which they share a high expression level of Kunitz-type inhibitors (KUNZ). On the other hand, comparative analyses revealed substantial differences in the expression levels of C-type lectins (CTL) and snake venom metalloproteinases (SVMP). The diverse set of SVMP and CTL isoforms shows signals of positive selection, and we also identified truncated forms of type III SVMPs, which resemble type II and type I SVMPs of viperids. Additionally, we identified a CNP precursor hosting a proline-rich region containing a BPP motif resembling those commonly detected in viperid venoms with hypotensive activity. Altogether, our results suggest an evolutionary history favoring high expression levels of few KUNZ isoforms in Phalotris venoms, contrasting with a highly diverse set of SVMP and CTL isoforms. Such diversity can be comparable with the venom variability observed in some viperids. Our findings highlight the extreme phenotypic diversity of non-front fanged snakes and the importance to allocate greater effort to study neglected groups of Colubroidea.

Assuntos

RESUMO

Although non-front fanged snakes account for almost two-thirds of snake diversity, most studies on venom composition and evolution focus exclusively on front-fanged species, which comprise most of the clinically relevant accidents. Comprehensive reports on venom composition of non-front fanged snakes are still scarce for several groups. In this study, we address such shortage of knowledge by providing new insights about the venom composition among species of Phalotris, a poorly studied Neotropical dipsadid genus. Phalotris are known for their specialized venom delivery system and toxic venoms, which can cause life-threatening accidents in humans. We evaluate the venom-gland transcriptome of Phalotris, comparing the following three South American species: P. reticulatus for the Araucaria Pine forests, P. lemniscatus for the Pampa grasslands, and P. mertensi for the Brazilian Cerrado. Our results indicate similar venom profiles, in which they share a high expression level of Kunitz-type inhibitors (KUNZ). On the other hand, comparative analyses revealed substantial differences in the expression levels of C-type lectins (CTL) and snake venom metalloproteinases (SVMP). The diverse set of SVMP and CTL isoforms shows signals of positive selection, and we also identified truncated forms of type III SVMPs, which resemble type II and type I SVMPs of viperids. Additionally, we identified a CNP precursor hosting a proline-rich region containing a BPP motif resembling those commonly detected in viperid venoms with hypotensive activity. Altogether, our results suggest an evolutionary history favoring high expression levels of few KUNZ isoforms in Phalotris venoms, contrasting with a highly diverse set of SVMP and CTL isoforms. Such diversity can be comparable with the venom variability observed in some viperids. Our findings highlight the extreme phenotypic diversity of non-front fanged snakes and the importance to allocate greater effort to study neglected groups of Colubroidea.

RESUMO

The bufonid species Melanophryniscus admirabilis is restricted to a single location in the southern Atlantic Forest, Brazil. Although the site of occurrence of M. admirabilis is covered with native forest and it is not directly exposed to pesticides application, the area is surrounded by agricultural activity. Our objectives were to evaluate possible alterations in morphological parameters (body mass, snout-vent length, and body index), metamorphosis (time to reach Gosner stages 42, 46 and to complete metamorphosis), and survival of M. admirabilis exposed to isolated Roundup® Original DI (R1: 234 and R2: 2340 µg.L-1 of glyphosate) and Boral® 500 SC, (B1: 130 and B2: 980 µg.L-1 of sulfentrazone) or mixed (R1+B1, R2+B1, R1+B2, R2+B2). Spawns of M. admirabilis were collected in natural lakes in the municipality of Arvorezinha and taken to laboratory cultivation. After the tadpoles acquired free swimming, the animals were acclimated for five days and fed ad libitum. The aquariums were contaminated with herbicides on the sixth day of cultivation, and the animals stayed in these aquariums for four days. Afterwards, the tadpoles were transferred to aquariums with clean water and monitored until metamorphosis (Gosner stage 46), when they were weighed, measured (snout-cloacal length) and cryoeuthanized. We observed no alterations in morphological parameters; however, survival was reduced in exposed groups (mortality index: 71 % in R2 and 29-64 % in mixed groups), suggesting energy allocation for metamorphosis at the expense of survival. Boral did not alter metamorphosis time. Roundup isolated and mixed with Boral altered the timing of Gosner stages 42 and 46 and reduced metamorphosis time, suggesting endocrine disruption. Thus, monitoring the presence and limiting the use of these pesticides in the area where M. admirabilis occurs can be crucial for conservation strategies.

Assuntos

RESUMO

Melanophryniscus admirabilis is a microendemic and critically endangered toad, known from a single population. This microendemic species inhabits a small fragment of the Atlantic Forest in South Brazil, an area significantly impacted by hydroelectric power plant projects, livestock farming, agricultural activities, biopiracy, and tourism. Given the exclusive and limited population of M. admirabilis, preserving and conserving this species is of utmost importance in Brazil. Research on this species primarily concentrates on its biology, ecology, and ecotoxicology. Currently, there is no knowledge about antimicrobial resistance (AMR) bacteria present in wild M. admirabilis, despite the potential for studying them to provide valuable insights into environmental pollution. To this end, Enterobacteriaceae species (n = 82) obtained from 15 wild M. admirabilis toads were subjected to the standard Kirby-Bauer disk diffusion method to test their AMR. The results showed that Enterobacteriaceae species had the highest antibiotic resistance to IPM (45.1%), CIP (39%), NIT (32.5%), AMP (31.3%), TET (18.3%), and FOX (17%). Of the tested species, 18 (21.9%) species tested were susceptible, 40 (48.8%) were resistant to 1 or 2 different antibiotic classes, and 24 (29.3%) were classified as multidrug-resistant. Overall, our findings suggest that the incidence of AMR in Enterobacteriaceae isolated from wild M. admirabilis is high, indicating environmental stress caused by anthropic pollution in their habit.

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RESUMO

Melanophryniscus admirabilis is a small toad, critically endangered with a microendemic distribution in the Atlantic Forest in southern Brazil. The amphibian skin microbiome is considered one of the first lines of defense against pathogenic infections, such as Batrachochytrium dendrobatidis (Bd). The knowledge of skin amphibian microbiomes is important to numerous fields, including species conservation, detection, and quantification of environmental changes and stressors. In the present study, we investigated, for the first time, cultivable bacteria in the skin of wild M. admirabilis, and detected Bd fungus by nested polymerase chain reaction (PCR) technique. Skin swab samples were collected from 15 wild M. admirabilis, and the isolation of bacteria was performed by means of different culture strategies. A total of 62 bacterial isolates being Bacillus (n = 22; 34.48%), Citrobacter (n = 10; 16.13%), and Serratia (n = 12; 19.35%) were more frequently isolated genera. Interestingly, all skin samples tested were Bd negative. Some bacterial genera identified in our study might be acting in a synergic relationship and protecting them against the Bd fungus. In addition, these bacteria may play an essential role in maintaining this species in an environment modulated by anthropic actions. This first report of skin cultivable bacteria from M. admirabilis natural population improves our knowledge of skin amphibian microbiomes, contributing to a better understanding of their ecology and how this species has survived in an environment modulated by anthropic action.

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RESUMO

Liotyphlops Peters, 1881 is an anomalepidid blindsnake genus that encompasses 12 species, widely distributed in Central and South America. In this study, we evaluate the taxonomic status of Liotyphlops sousai Santos & Reis, 2018, a species described based on a single specimen from the Atlantic Rainforest of southern Brazil, and after analyzing evidences based upon molecular systematics, external morphology and osteology, we propose its synonymy with Liotyphlops beui (Amaral, 1924), a common and widely distributed Anomalepidid species. We also provide comments and recommendations on the taxonomy of Liotyphlops, assessing the risks associated with describing a new species based on small type series.

RESUMO

Melanophryniscus admirabilis (admirable red-belly toad) is a microendemic and critically endangered species found exclusively along 700 m of the Forqueta River, in a fragment of the Atlantic Forest of southern Brazil. One of the greatest concerns regarding the conservation of this species is the extensive use of pesticides in areas surrounding their natural habitat. In recent years, the adaptation and persistence of animal species in human-impacted environments have been associated with microbiota. Therefore, the present study aimed to characterize the oral bacterial community of wild M. admirabilis and to address the question of how this community might contribute to this toad's adaptation in the anthropogenic environment as well as its general metabolic capabilities. A total of 11 oral samples collected from wild M. admirabilis were characterized and analyzed via high-throughput sequencing. Fragments of the 16S rRNA variable region 4 (V4) were amplified, and sequencing was conducted using an Ion Personal Genome Machine (PGM) System with 316 chips. A total of 181,350 sequences were obtained, resulting in 16 phyla, 34 classes, 39 orders, and 77 families. Proteobacteria dominated (53%) the oral microbiota of toads, followed by Firmicutes (18%), Bacteroidetes (17%), and Actinobacteria (5%). No significant differences in microbial community profile from among the samples were reported, which suggests that the low dietary diversity observed in this population may directly influence the bacterial composition. Inferences of microbiome function were performed using PICRUSt2 software. Important pathways (e.g., xenobiotic degradation pathways for pesticides and aromatic phenolic compounds) were detected, which suggests that the bacterial communities may serve important roles in M. admirabilis health and survival in the anthropogenic environment. Overall, our results have important implications for the conservation and management of this microendemic and critically endangered species.

RESUMO

Melanophryniscus admirabilis is a frog endemic to the southern Atlantic Forest (Brazil), with restricted distribution and considered as critically endangered. The aim of this study was to evaluate possible alterations in biomarkers of metabolism (glycogen, proteins, and uric acid) and oxidative balance (superoxide dismutase, catalase, glutathione S-transferase, and lipoperoxidation) of tadpoles of Melanophryniscus admirabilis exposed to commercial herbicide formulations containing sulfentrazone (Boral® 500 SC: 130 and 980 µg a.i./L) and glyphosate (Roundup® Original: 234 and 2340 µg a.i./L). Mortality was not observed in any of the groups studied. Our results show that a 96-h exposure to the herbicides decreased glycogen levels, indicating increased energy demand for xenobiotic metabolism. Protein levels increased in the Boral group but decreased in the higher concentration of Roundup, and uric acid levels did not change significantly between the experimental groups. Lipoperoxidation decreased in the Boral group and in the higher concentration of Roundup. Decreased levels of superoxide dismutase in both treatments and of catalase in the lowest concentration of the herbicides were observed. Glutathione S-transferase activity increased in the Roundup group; this enzyme seems to be crucial in the metabolization of the herbicides and in the survival of the tadpoles. Our results suggest that M. admirabilis has a high antioxidant capacity, which guaranteed the survival of tadpoles. Nevertheless, exposure to pesticides could impose a serious risk to this species, especially considering its restricted distribution, habitat specificity, and high physiological demand to metabolize xenobiotics.

Assuntos

RESUMO

In anuran amphibians, acoustic signals are fundamental mechanisms of mate recognition and mate choice, which makes frog calls a fundamental tool for anuran taxonomy. In this work, we describe the advertisement call of two species for the genus Melanophryniscus, M. cambaraensis and M. macrogranulosus and use the descriptions to try to solve a taxonomic problem between them. We collected data after heavy rains in three different sample sites in Rio Grande do Sul, Brazil, between 2012 and 2013. The advertisement call of both species is composed of two segments. It always begins with part A (about 0.44-6 seconds) composed of single modulated pulses separated by long time intervals. It is followed by part B, a long train of unmodulated pulses with short time intervals, lasting from 9 to 32.2 seconds. Principal Component Analysis (PCA) indicated some variation between temporal parameters of the two species, but Multivariate Analysis of Variance showed no significant differences. Within-individual Coefficient of Variation (CV) showed only two static parameters: pulse rate and peak frequency, both in the part B of the call. Despite intra-male variation in some acoustic parameters, it is not possible to differentiate between M. cambaraensis and M. macrogranulosus species only using bioacoustics.

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