Insights into the evolution of photoreceptor oil droplets in frogs and toads.

Photoreceptor oil droplets (ODs) are spherical organelles placed most commonly within the inner segment of the cone photoreceptors. Comprising neutral lipids, ODs can be either non-pigmented or pigmented and have been considered optically functional in various studies. Among living amphibians, ODs were only reported to occur in frogs and toads (Anura), while they are absent in salamanders and caecilians. Nonetheless, the limited understanding of their taxonomic distribution in anurans impedes a comprehensive assessment of their evolution and relationship with visual ecology. We studied the retinae of 134 anuran species, extending the knowledge of the distribution of ODs to 46 of the 58 currently recognized families, and providing a new perspective on this group that complements the available information from other vertebrates. The occurrence of ODs in anurans shows a strong phylogenetic signal, and our findings revealed that ODs evolved at least six times during the evolutionary history of the group, independently from other vertebrates. Although no evident correlation was found between OD occurrence, adult habits and diel activity, it is inferred that each independent origin involves distinct scenarios in the evolution of ODs concerning photic habits. Furthermore, our results revealed significant differences in the size of the ODs between nocturnal and arrhythmic anurans relative to the length of the cones' outer segment.

Transcription factor roles in the local adaptation to temperature in the Andean Spiny Toad Rhinella spinulosa.

Environmental temperature strongly influences the adaptation dynamics of amphibians, whose limited regulation capabilities render them susceptible to thermal oscillations. A central element of the adaptive strategies is the transcription factors (TFs), which act as master regulators that orchestrate stress responses, enabling species to navigate the fluctuations of their environment skillfully. Our study delves into the intricate relationship between TF expression and thermal adaptation mechanisms in the Rhinella spinulosa populations. We sought to elucidate the dynamic modulations of TF expression in prometamorphic and metamorphic tadpoles that inhabit two thermally contrasting environments (Catarpe and El Tatio Geyser, Chile) and which were exposed to two thermal treatments (25 °C vs. 20 °C). Our findings unravel an intriguing dichotomy in response strategies between these populations. First, results evidence the expression of 1374 transcription factors. Regarding the temperature shift, the Catarpe tadpoles show a multifaceted approach by up-regulating crucial TFs, including fosB, atf7, and the androgen receptor. These dynamic regulatory responses likely underpin the population's ability to navigate thermal fluctuations effectively. In stark contrast, the El Tatio tadpoles exhibit a more targeted response, primarily up-regulating foxc1. This differential expression suggests a distinct focus on specific TFs to mitigate the effects of temperature variations. Our study contributes to understanding the molecular mechanisms governing thermal adaptation responses and highlights the resilience and adaptability of amphibians in the face of ever-changing environmental conditions.

Runway extinction in terrestrial toads (Rhinella arenarum): Instrumental or Pavlovian?

Instrumental appetitive extinction involves the reduction of a previously reinforced response when its occurrence is no longer rewarded. Two experiments with terrestrial toads (Rhinella arenarum) tested whether the occurrence of a nonreinforced response is necessary for response extinction by varying the time of exposure to nonrewarded goal-box stimuli across groups. In Experiment 1, toads that received the same acquisition training (15 sessions, 1 session/day, 300 s of access to water in the goal box) were randomly assigned to two groups. In Group 600 (n=12), animals spent 600 s in the goal box in 8 daily extinction sessions (water present but inaccessible). In Group 0 (n=11), toads performed the runway response (i.e., walking from the start to the goal box) but were removed as soon as they entered the goal box, thus having minimal exposure to nonrewarded goal-box stimuli. The runway response was weakened in Group 600 across extinction trials, but exhibited little change in Group 0. In Experiment 2, toads were randomly assigned to two groups after the same acquisition training. Group 0 (n=7) was treated the same as Group 0 in the previous experiment. In Group RI (retention interval, n=7), toads remained in their home cage for 13 days. Finally, all animals received 4 extinction sessions with 300 s in the empty goal box. There was little behavioral change in Group 0 during the 13 sessions with minimal exposure to the goal box. In extinction, both groups reduced their runway response at similar rates. Although the procedures were instrumental, extinction of the runway response in toads can be accounted for in terms of a Pavlovian approach response to stimuli paired with reward and nonreward in the goal box.

The Impacts of Transdermal Application of Corticosterone on Toad (Rhinella icterica) Immunity.

Recent studies have shown that acute physiological increases in endogenous glucocorticoid levels have immunostimulatory effects. Although post-acute stress immunosuppressive effects have also been described, the difference between enhancing and suppressing the immune response seems mediated by the stressor's duration, intensity, and the immune component under analysis. To elicit physiologically relevant corticosterone levels that can be found in Rhinella icterica toads after stressful events (e.g., restraint or captivity) and understand how acute increased glucocorticoid levels of different intensities affect corticosterone and testosterone plasma levels and immune parameters (in vitro plasma bacterial killing ability, neutrophil-to-lymphocyte ratio, and in vivo phagocytosis of peritoneal leukocytes), we submitted toads to the transdermal application of two corticosterone doses (1 and 10 µg). Corticosterone transdermal application increased corticosterone plasma levels with different intensities: 3 times for 1 µg and fourteen times for 10 µg, compared to the vehicle, and the neutrophil-to-lymphocyte ratio increased regardless of the corticosterone dose. However, there was no effect on testosterone levels and bacterial killing ability. Interestingly, both corticosterone doses promoted immunosuppression, decreasing peritoneal leukocytes' phagocytosis activity by 60% for toads receiving the dose of 1µg and 40% for those receiving 10 µg. Our results show the complexity of the relationship between increased corticosterone levels and immunomodulation. The different corticosterone doses promoted increases of distinct magnitudes in corticosterone plasma levels, with the less intense increase in corticosterone levels generating greater cell-mediated immunosuppression. Future studies using different corticosterone doses to achieve and compare physiological vs. pharmacological hormone levels are imperative to understanding these interrelationships between corticosterone and immune response.

Increased rates of dispersal of free-ranging cane toads (Rhinella marina) during their global invasion.

Invasions often accelerate through time, as dispersal-enhancing traits accumulate at the expanding range edge. How does the dispersal behaviour of individual organisms shift to increase rates of population spread? We collate data from 44 radio-tracking studies (in total, of 650 animals) of cane toads (Rhinella marina) to quantify distances moved per day, and the frequency of displacement in their native range (French Guiana) and two invaded areas (Hawai'i and Australia). We show that toads in their native-range, Hawai'i and eastern Australia are relatively sedentary, while toads dispersing across tropical Australia increased their daily distances travelled from 20 to 200 m per day. That increase reflects an increasing propensity to change diurnal retreat sites every day, as well as to move further during each nocturnal displacement. Daily changes in retreat site evolved earlier than did changes in distances moved per night, indicating a breakdown in philopatry before other movement behaviours were optimised to maximise dispersal.

Feeding alters the preferred body temperature of Cururu toads, Rhinella diptycha (Anura, Bufonidae).

Ectothermic organisms depend primarily on external heat sources and behavioural adjustments to regulate body temperature. Under controlled conditions, in a thermal gradient, body temperature often clusters around a more or less defined range of preferred body temperatures (Tpref). However, Tpref may be modified in response to environmental parameters and/or physiological state. For example, meal ingestion is sometimes followed by a post-prandial thermophilic response leading to a transient increment in Tpref. Although thought to optimize digestive processes, its occurrence, magnitude, and possible determinants remains scarcely documented for anuran amphibians. Herein, we investigated whether the Cururu toad, Rhinella diptycha, exhibits a post-prandial thermophilic response by monitoring the body temperature of fasting and fed toads while they were maintained in a thermal gradient. We found that the toads' Tpref increased by about 13% from day 2 to 4 after feeding, in comparison with the Tpref recorded under fasting. Also, fed animals exhibited a broader range for Tpref at days 2 and 3 post-prandial, which reflects a greater level of locomotor activity compared to fasting individuals. We conclude that R. diptycha is capable to exhibit a post-prandial thermophilic response under the controlled conditions of a thermal gradient. Although this thermoregulatory adjustment is thought to optimize meal digestion yielding important energetic and ecological benefits, its occurrence in anuran amphibians in nature remains uncertain.

Toads prey upon scorpions and are resistant to their venom: A biological and ecological approach to scorpionism.

In recent years, SE Brazil, the most populous region in the country with an estimated population of 88 million, has been experiencing an alarming increase in scorpions accidents (scorpionism), mainly caused by the yellow scorpion (Tityus serrulatus), or "escorpião amarelo" in Portuguese. This species is considered particularly dangerous to humans and can reproduce by parthenogenesis favouring rapid dispersal and colonization of new environments. Since the 1940s, owing to the growing danger represented by scorpionism, public control policies have been developed, including active search for scorpions, together with the use of toxic substances applied in places most likely to serve as their refuges. Even so, the number of accidents is increasing year by year, presently at an alarming rate. It seems evident that the increase in accidents is directly (or primarily) related to the lack of predators that in healthy environmental conditions would naturally control scorpion populations. However, due to environmental changes, leading to a lack of predators, scorpions have been gradually invading the urban environment. Arachnids and insects in general, as well as some other invertebrates, are preyed upon by anuran amphibians (toads, frogs and tree frogs). Toads (family Bufonidae) are nocturnal, large, and highly voracious animals, capable of actively exploring extensive areas and consuming large numbers of insects and arachnids daily. One of the most common toad species in southeastern Brazil is Rhinella icterica. Both R. icterica and T. serrulatus inhabit the same nocturnal environment. The predatory action of toads, specifically on scorpions, is practically unknown from behavioural and toxinological points of view. Thus, we studied the predatory behaviour of this toad against the yellow scorpion and evaluated the resistance of the amphibian to scorpion venom. Our results show that R. icterica is a voracious predator of T. serrulatus and is extremely resistant to its venom. Human/toad relationship throughout western history has always been very conflicted and possibly one of the factors that most has contributed to human ignorance of the role of these amphibians in maintaining ecological balance. Presently, the control of scorpionism is being performed through active search and/or the use of chemical agents, although showing little efficacy in reducing human accidents. In the medium or long term, more effective actions taking into account the biology of scorpions and their predators have never been taken to reduce these accidents.

Predation by non-bioluminescent firefly larvae on a tepui-summit endemic toad.

Tepuis are Precambrian sandstone tabletop mountains in South America that can reach up to ∼3,000 m in elevation. Their highest summits are both physiographically and ecologically isolated from the surrounding upland savannah and lush tropical rainforest, and they face particularly hostile, challenging environmental conditions [1,2] (Figure 1A). Taxa thriving on high tepui summits must adapt to resource-limited and highly competitive ecosystems. The toad genus Oreophrynella is exclusively found on tepui slopes and summits [3] and is particularly well-adapted to the tepui top environment. We here report on an unanticipated and possibly large-scale predation on this endemic toad by non-bioluminescent firefly larvae.

Time-related immunomodulation by stressors and corticosterone transdermal application in toads.

Immune responses have been mostly studied at a specific time in anuran species. However, time-changes related to immunomodulation associated with glucocorticoid (GC) alterations following stressors and GC treatment are complex. The present study describes time-related changes in immune response and corticosterone (CORT) plasma levels following restraint challenge, short, mid and long-term captivity, and CORT exogenous administration by transdermal application (TA) in Rhinella ornata toads. We observed increased neutrophil: lymphocyte ratios after restraint challenge and CORT TA, without changes following short and mid-term captivity. Plasma bacterial killing ability was sustained in all treatments, except long-term captivity, with decreased values after 90 days under such conditions. Phagocytic activity of peritoneal cells increased after mid-term captivity, and the phytohemagglutinin swelling response was impaired in those animals treated with CORT TA for 20 consecutive days. Plasma CORT levels increased or were sustained after restraint challenge (depending on initial values), decreased following mid and long-term captivity (for those animals showing high CORT in the field) and increased after 20 days of CORT TA. By performing assessments of time-changes in immune processes and CORT plasma levels in R. ornata, we demonstrate immuno-enhancing effects following restraint, short and mid-term stressors, while long-term stressors and CORT TA promoted immunosuppression in these toads.

The advertisement call of the phytotelm-breeding Melanophryniscus xanthostomus (Anura: Bufonidae)

Vocalizations are often useful for understanding taxonomic relationships among anuran species. Despite this usefulness, vocalizations are described in only nine of 29 in Melanophryniscus Gallardo, 1961. Here we describe the advertisement call of Melanophryniscus xanthostomus Baldo, Bornschein, Pie, Ribeiro, Firkowski & Morato, 2015 of a population from Serra Dona Francisca, municipality of Campo Alegre, state of Santa Catarina, Brazil. The advertisement call (of three males, total of 17 calls) comprises two segments (the first with short and single notes, followed by a multi-pulsed note), with a duration of 12.194-20.986 s, and dominant frequency of 3101-3618 Hz (first and second segments combined). The advertisement call of M. xanthostomus differs from its congeners mostly by the higher number of pulses in the second segment (294-1033; from 16 to 321 in the other Melanophryniscus species), except Melanophryniscus krauczuki Baldo & Basso, 2004 (1018-1502 pulses in the second segment). This is the first described call of a phytotelm breeding Melanophryniscus, but it presents the same prototype (a compound call formed by two segments, the first composed of short notes followed by a long trill) of its congeners not-phytotelm breedings. This might indicate the advertisement call of Melanophryniscus as a conserved trait and thus can be considered diagnostic for the genus.(AU)

Differences in ocular media transmittance in classical frog and toad model species and its impact on visual sensitivity.

The transmittance properties of the cornea, lens and humours of vertebrates determine how much light across the visible spectrum reaches the retina, influencing sensitivity to visual stimuli. Amphibians are the only vertebrate class in which the light transmittance of these ocular media has not been thoroughly characterised, preventing large-scale comparative studies and precise quantification of visual stimuli in physiological and behavioural experiments. We measured the ocular media transmittance in some commonly used species of amphibians (the bufonids Bufo bufo and Rhinella ornate, and the ranids Lithobates catesbeianus and Rana temporaria) and found low transmittance of short wavelength light, with ranids having less transmissive ocular media than bufonids. Our analyses also show that these transmittance properties have a considerable impact on spectral sensitivity, highlighting the need to incorporate this type of measurement into the design of stimuli for experiments on visual function.

Evaluation of antimutagenic and cytotoxic activity of skin secretion extract of Rhinella marina and Rhaebo guttatus (Anura, Bufonidae) / Avaliação da atividade antimutagênica e citotóxica da secreção cutânea de Rhinella marina e Rhaebo guttatus (Anura, Bufonidae)

The skin secretion from toads of the Bufonidae family has great potential in the search for new active compounds to be used as drug candidates in treating some diseases, among them cancer. In this context, this study aimed to evaluate the cytotoxic and antimutagenic activity of the parotoid gland secretion extracts of Rhinella marina and Rhaebo guttatus, as well as biochemically analyze transaminases and serum creatinine for liver and renal damage, respectively. Cytotoxicity was performed by the colorimetric method based on MTT (3- [4, 5-dimethyl-2-thiazolyl]-2, 5-diphenyl-2H-tetrazolium bromide) with different concentrations of the extracts in Walker or splenic tumor cell cultures from rats and mice. The micronucleus test was performed with male Swiss mice treated orally with the extracts for 15 days, and then intraperitoneally with N-ethyl-N-nitrosurea (50 mg kg-1). Micronucleated polychromatic erythrocytes (MNPCE) were evaluated in bone marrow. The extracts showed cytotoxic activity in the evaluated cells. There was a significant reduction in the frequency of MNPCE (R. marina = 56% and R. guttatus = 75%, p < 0.001), indicating antimutagenic potential of the extracts. The groups treated only with extract showed an increase in MNPCE frequency, evidencing mutagenic potential. Biochemical analyzes showed no significant difference between treatments. Thus, under our experimental conditions, the extracts of R. marina and R. guttatus skin secretions presented chemopreventive potential for cancer. (AU)

A secreção cutânea de anuros da família Bufonidae tem grande potencial na busca de novos compostos ativos para utilização como fármacos candidatos no tratamento de algumas doenças, entre elas o câncer. Neste contexto, este estudo teve como objetivo avaliar a atividade citotóxica e antimutagênica dos extratos da secreção da glândula parótida de Rhinella marina e Rhaebo guttatus, bem como a análise bioquímica de transaminases e creatinina séricas, para avaliar dano hepático e renal, respectivamente. A avaliação de citotoxicidade foi realizada pelo método colorimétrico baseado no MTT (3-[4, 5-dimethyl-2-thiazolyl]-2, 5-diphenyl-2H-tetrazolium bromide), com diferentes concentrações dos extratos em culturas de células do Tumor de Walker ou células esplênicas de rato e camundongo. O teste do micronúcleo foi realizado com camundongos Swiss machos que receberam tratamento oral com os extratos durante 15 dias, seguido de tratamento intraperitoneal com N-etil-N-nitrosuréia (50 mg kg-1). A frequência de eritrócitos policromáticos micronucleados (PCEMN) foi determinada em medula óssea. Os extratos apresentaram ação citotóxica nas células avaliadas. Houve uma redução significativa na frequência de PCEMN (R. marina = 56% e R. guttatus = 75%, p < 0,001), observando-se um potencial antimutagênico dos extratos. Os grupos tratados somente com os extratos apresentaram um aumento na frequência de PCEMNs, evidenciando um potencial mutagênico. As análises bioquímicas não apresentaram diferença significativa entre os tratamentos. Assim, nas condições experimentais testadas, as secreções cutâneas de R. marina e R. guttatus apresentaram potencial quimiopreventivo para câncer.(AU)

The advertisement call of the phytotelm-breeding Melanophryniscus xanthostomus (Anura: Bufonidae)

Vocalizations are often useful for understanding taxonomic relationships among anuran species. Despite this usefulness, vocalizations are described in only nine of 29 in Melanophryniscus Gallardo, 1961. Here we describe the advertisement call of Melanophryniscus xanthostomus Baldo, Bornschein, Pie, Ribeiro, Firkowski & Morato, 2015 of a population from Serra Dona Francisca, municipality of Campo Alegre, state of Santa Catarina, Brazil. The advertisement call (of three males, total of 17 calls) comprises two segments (the first with short and single notes, followed by a multi-pulsed note), with a duration of 12.194-20.986 s, and dominant frequency of 3101-3618 Hz (first and second segments combined). The advertisement call of M. xanthostomus differs from its congeners mostly by the higher number of pulses in the second segment (294-1033; from 16 to 321 in the other Melanophryniscus species), except Melanophryniscus krauczuki Baldo & Basso, 2004 (1018-1502 pulses in the second segment). This is the first described call of a phytotelm breeding Melanophryniscus, but it presents the same prototype (a compound call formed by two segments, the first composed of short notes followed by a long trill) of its congeners not-phytotelm breedings. This might indicate the advertisement call of Melanophryniscus as a conserved trait and thus can be considered diagnostic for the genus.

Artificial light at night and captivity induces differential effects on leukocyte profile, body condition, and erythrocyte size of a diurnal toad.

Light pollution or artificial lighting at night (ALAN) is an emerging threat to biodiversity that can disrupt physiological processes and behaviors. Because ALAN stressful effects are little studied in diurnal amphibian species, we investigated if chronic ALAN exposure affects the leukocyte profile, body condition, and blood cell sizes of a diurnal toad. We hand-captured male toads of Melanophryniscus rubriventris in Angosto de Jaire (Jujuy, Argentina). We prepared blood smears from three groups of toads: "field" (toads processed in the field immediately after capture), "natural light" (toads kept in the laboratory under captivity with natural photoperiod), and "constant light" (toads kept in the laboratory under captivity with constant photoperiod/ALAN). We significantly observed higher neutrophil proportions and neutrophils to lymphocytes ratio in toads under constant light treatment. In addition, we observed significantly better body condition and higher erythrocyte size in field toads compared with captive toads. In summary, ALAN can trigger a leukocyte response to stress in males of the diurnal toad M. rubriventris. In addition, captivity can affect the body condition and erythrocyte size of these toads.

Dehydration as a stressor in toads (Rhinella ornata).

Availability of a humid environment is essential for amphibians to carry out their activities and most species are characterized by low resistance to evaporative water loss. Moreover, dehydration severely compromises amphibian locomotor and foraging performance, representing a relevant selective factor modulating the evolution of its integrative phenotype. In this way, we hypothesized that dehydration is a stressor for toads, inducing a stress response comparable to that elicited by another commonly used stress protocol: restraint challenge. We evaluated changes in plasma levels of corticosterone (CORT), hematocrit (Hct), and neutrophil:lymphocyte (N:L) ratio in adult males of Rhinella ornata, experimentally submitted to different levels of hydration (100%, 90%, and 80% of standard body mass) and to restraint challenge. Our results showed that dehydrating toads by 10% increase CORT to levels equivalent to that obtained by restraint. Moreover, toads dehydrated by 20% show a more pronounced increase in CORT, along with increased Hct and N:L ratio. In this way, we corroborated the hypothesis that dehydration triggers a pronounced stress response in R. ornata.

Interplay among steroids, body condition and immunity in response to long-term captivity in toads.

Stressful experiences can promote harmful effects on physiology and fitness. However, stress-mediated hormonal and immune changes are complex and may be highly dependent on body condition. Here, we investigated captivity-associated stress effects, over 7, 30, 60, and 90 days on plasma corticosterone (CORT) and testosterone (T) levels, body index, and innate immunity (bacterial killing ability and phagocytosis of peritoneal cells) in toads (Rhinella icterica). Toads in captivity exhibited elevated CORT and decreased T and immunity, without changes in body index. The inter-relationships between these variables were additionally contrasted with those obtained previously for R. schneideri, a related species that exhibited extreme loss of body mass under the same captive conditions. While T and phagocytosis were positively associated in both species, the relationship between CORT and bacterial killing ability was dependent on body index alterations. While CORT and bacterial killing ability were positively associated for toads that maintained body index, CORT was negatively associated with body index in toads that lost body mass over time in captivity. In these same toads, body index was positively associated with bacterial killing ability. These results demonstrate that steroids-immunity inter-relationships arising from prolonged exposure to a stressor in toads are highly dependent on body condition.

Temperature and dehydration effects on metabolism, water uptake and the partitioning between respiratory and cutaneous evaporative water loss in a terrestrial toad.

Terrestrial anurans often experience fluctuations in body temperature and hydration state, which are known to influence evaporative water loss through the skin (EWLSkin) and lungs (EWLResp). These effects arise from associated changes in skin permeability, metabolism and lung ventilation. Herein, we determined the rates of EWLSkin and EWLResp in the terrestrial toad Rhinella diptycha at different temperatures and hydration states. We measured oxygen uptake rates to verify whether alterations in the partitioning between EWLSkin and EWLResp were associated with metabolism-induced changes in pulmonary gas exchange. We also measured the influence of hydration and temperature on water uptake (WU) through the skin. Finally, as estimates of skin resistance to evaporation (Rs) are usually inferred from total evaporative water loss (EWLTotal), under the assumption of negligible EWLResp, we calculated the potential error in accepting this assumption for different temperature and hydration states. EWLSkin and EWLResp increased with temperature, but this response was greater for EWLResp, which was attributed to the temperature-induced elevation in metabolism and lung ventilation. Dehydration caused a decrease in the relative contribution of EWLSkin to EWLTotal, mirrored by the concurrent increase in the contribution of EWLResp, at all temperatures. Thus, Rs increased with dehydration. WU rates were dictated by dehydration with little influence of temperature. The partitioning between EWLSkin and EWLResp was affected by both temperature and hydration state and, under some conditions, considering EWLResp as negligible led to significant errors in the assessment of skin resistance to evaporation.

Unraveling fine-scale habitat use for secretive species: When and where toads are found when not breeding.

A good understanding of species-habitat associations, or habitat use, is required to establish conservation strategies for any species. Many amphibian species are elusive and most information concerning amphibian habitat use comes from breeding sites where they are comparatively easy to find and study. Knowledge about retreat sites is extremely limited for most species and for the greater part of the year. For such species, it is especially important to factor in detection probability in habitat analyses, because otherwise distorted views about habitat preferences may result, e.g., when a species is more visible in habitat type B than in A, even though A may be preferred. The South American red-belly toad, Melanophryniscus pachyrhynus, is a range-restricted species from Southern Brazil and Uruguay that inhabits open areas with rocky outcrops and is usually seen only during explosive breeding events. Here we studied the fine-scale habitat use of the red-belly toad outside of the breeding season to identify retreat sites and test for the importance of accounting for species imperfect detection, using Bayesian occupancy models. We identified shrub density and the number of loose rocks as important predictors of occupancy, while detection probability was highest at intermediate temperatures. Considering the harsh (dry and hot) conditions of rocky outcrops, shrubs and loose rocks may both work as important refuges, besides providing food resources and protecting against predation. Rocky outcrops have been suffering changes in habitat configuration and we identify nonbreeding habitat preferences at a fine scale, which may help to promote population persistence, and highlight the importance of accounting for imperfect detection when studying secretive species.

Entomotoxic activity of Rhinella icterica (Spix, 1824) toad skin secretion in Nauphoeta cinerea cockroaches: An octopamine-like modulation.

Rhinella icterica is a poisonous toad whose toxic secretion has never been studied against entomotoxic potential. Sublethal doses of Rhinella icterica toxic secretion (RITS) were assayed in Nauphoeta cinerea cockroaches, in order to understand the physiological and behavioral parameters, over the insect central and peripheral nervous system. RITS (10 µg/g) injections, induced behavioral impairment as evidenced by a significant decrease (38 ±â€¯14%) in the distance traveled (p < .05), followed by an increase (90 ±â€¯6%) of immobile episodes (p < .001, n = 28, respectively). In cockroaches semi-isolated heart preparations, RITS (16 µg/200 µl) induced a significant irreversible dose-dependent negative chronotropism, reaching ~40% decrease in heart rate in 20 min incubation. In in vivo cockroach neuromuscular preparations, RITS (20, 50 and 100 µg/g of animal weight) induced a time-dependent inhibition of twitch tension that was complete for 20 µg/g, in 120 min recordings. RITS (10 µg/g) also induced a significant increase in the insect leg grooming activity (128 ±â€¯10%, n = 29, p < .01), but not in the antennae counterparts. The RITS increase in leg grooming activity was prevented in 90% by the pretreatment of cockroaches with phentolamine (0.1 µg/g). The electrophysiological recordings of spontaneous neural compound action potentials showed that RITS (20 µg/g) induced a significant increase in the number of events, as well as in the rise time and duration of the potentials. In conclusion, RITS showed to be entomotoxic, being the neuromuscular failure and cardiotoxic activity considered the main deleterious effects. The disturbance of the cockroaches' behavior together with the electrophysiological alterations, may unveil the presence of some toxic components present in the poison with inherent biotechnological potentials.