Toxic tasting: how capuchin monkeys avoid grasshoppers' chemical defenses.

Platyrrhines consume many species of arthropods in the order Orthoptera. Some species of orthopterans can produce chemical defenses that render them toxic or unpalatable and thus act as predator deterrents. These species include the stick grasshoppers (family Proscopiidae), which are widely distributed in the Caatinga biome in northeastern Brazil, which comprises part of the distribution of capuchin monkeys. Capuchin monkeys are omnivores and consume a wide variety of foods, including unpleasant-tasting, potentially toxic items, which they need to learn how to process. We describe the processing of stick grasshoppers (Stiphra sp.) by wild capuchin monkeys (Sapajus libidinosus) that live in Serra da Capivara National Park, Brazil, and compare how individuals of different age classes handle these potentially toxic food items. S. libidinosus predominantly avoided consuming the digestive tract, which contains toxic compounds, when feeding on stick grasshoppers. Immatures took longer than adults to process the stick grasshoppers, indicating that capuchins need to learn how to process the toxic digestive tract of these prey to avoid consuming it.

Role of ABC Proteins in Secondary Metabolism and Immune (=Defensive) Response in Seaweeds.

Laurencia seaweed species synthesize a broad range of secondary metabolites, mainly terpenes (e.g., elatol), exhibiting diverse ecological roles, such as defense against fouling and herbivores. Recently, an intricate cellular machinery was described concerning terpenes biosynthetic pathways, storage inside corps en cerise (CC), and regulated exocytosis in these species. But for seaweeds in general, the proteins involved in transmembrane transport of secondary metabolites remain unknown. Assays with Rhodamine-123 and cyclosporine A (CSA) revealed the presence of ABC transporters in CC membrane of Laurencia dendroidea. In vivo incubation assays with CSA resulted in CC morphological changes, reduced intracellular elatol concentrations, and increased biofouling cover on the seaweed surface. Cultivation assays in the presence of a marine pathogenic bacteria induced the expression of ABC proteins belonging to the subfamilies ABCB, ABCD, ABCF, and ABCG. The latter subfamily is known to be associated with the transport of plant terpenes. Our results shed new light on the role of ABC proteins in key mechanisms of the defensive system in seaweeds against fouling and herbivory.

Which variables influence the herbivory amount on Montrichardia spp. (Araceae) in aquatic ecosystems? / ¿Qué variables influyen en la herbivoría en Montrichardia spp. (Araceae) en los ecosistemas acuáticos?

Abstract Introduction: The study of herbivory is fundamental in ecology and includes how plants invest in strategies and mechanisms to reduce herbivore damage. However, there is still a lack of information about how the environment, plant density, and functional traits influence herbivory in aquatic ecosystems. Objective: To assess if there is a relationship between herbivory, environmental variables, and plant traits two species of Montrichardia, a neotropical aquatic plant. Methods: In September 2018, we studied 78 specimens of Montrichardia arborescens and 18 of Montrichardia linifera, in 18 sites in Melgaço, Pará, Brazil. On each site, we measured water depth, distance to the margin, and plant density. From plants, we measured plant height and leaf thickness, and photographed the leaves to calculate the specific leaf area and percentage herbivory. To identify anatomical structures, we collected fully expanded leaves from three individuals per quadrat. Results: For M. arborescens, plants with thicker leaves and higher specific leaf area have less herbivore damage. For M. linifera, plants from deeper sites and with thicker leaves had more herbivore damage, while plants that grew farther from the margin had less damage. We found anatomical structures related to defense, such as idioblast cells with phenolic compounds, and cells with solid inclusions that can contribute to avoiding severe damage. Conclusions: Herbivory in these Montrichardia species can be explained by a combination of plant and environmental traits (patch isolation and water depth). The main plant traits are leaf thickness and area, but chemical compounds and solid inclusions also help Montrichardia to sustain less damage than other macrophytes.

Resumen Introducción: La herbivoría es fundamental para comprender cómo las plantas invierten en diferentes estrategias para evitar la depredación, lo que implica diferentes mecanismos de defensa. Factores relacionados con el medio ambiente, la densidad de plantas y/o los rasgos funcionales de las plantas pueden influir en la herbivoría en los ecosistemas acuáticos. Sin embargo, todavía falta información sobre cómo esos factores influyen en la herbivoría en los ecosistemas acuáticos y pueden contribuir a la carga de herbivoría. Objetivo: Evaluar si existe una relación entre la herbivoría y las variables ambientales (p. ej., profundidad del agua y distancia al margen), los factores ecológicos (densidad de plantas) y los rasgos estructurales de las plantas (altura, grosor de la hoja y área foliar) e indicar estructuras anatómicas que actúen junto con los rasgos estructurales en el sistema de defensa de especies de Montrichardia. Métodos: Se evaluaron 96 individuos de Montrichardia spp. (78 de M. arborescens y 18 de M. linifera, en 18 sitios) recolectados en septiembre de 2018. En cada sitio, se midió la profundidad del agua, la distancia al margen y la densidad de plantas. De los individuos, medimos la altura de la planta, el grosor de la hoja y fotografiamos las hojas para calcular el área foliar específica y la cantidad de herbivoría (en porcentaje). Para identificar las estructuras anatómicas relacionadas con la defensa de las plantas, se recogió hojas completamente expandidas de tres individuos por cuadrante. Resultados: Para M. arborescens, las plantas con hojas más gruesas y mayor área foliar específica tienen menos daño por herbivoría. Para M. linifera, las plantas con hojas más gruesas y que habitan en sitios más profundos tienen más daño por herbívoros, mientras que las plantas más alejadas del margen tienen menos daño por herbívoros. Se encontró estructuras anatómicas relacionadas con la defensa, como células idioblásticas con compuestos fenólicos y células con inclusiones sólidas que pueden contribuir a evitar daños severos en las características de las hojas. Conclusiones: Nuestros resultados indican que la herbivoría en las especies de Montrichardia podría explicarse por una combinación de características ambientales (aislamiento del parche y profundidad del agua) y de la planta. Descubrimos que los rasgos de las hojas eran factores importantes que impulsaban los cambios en la carga de herbivoría, especialmente el grosor de las hojas y el área foliar específica. Además, las especies de Montrichardia invierten en compuestos químicos e inclusiones sólidas para evitar daños graves en las hojas y, por lo tanto, pueden sufrir menos daños que otras especies de macrófitos.

Meta-analysis of tadpole taste tests: consumption of anuran prey across development and predator strategies.

The risk of predation and the costs and benefits of diverse anti-predator strategies can shift across the life stages of an organism. Yet, empirical examples of ontogenetic switches in defense mechanisms are scarce. Anurans represent an alleged exception; previous meta-analytic work suggests that unpalatability of developing anurans is "rare", whereas adult anurans in many lineages are well defended by toxic and/or unpalatable skin secretions. Here, we revisit the question of the unpalatability of anuran young in a meta-analysis of the relative proportion of prey consumed within 922 predation tests, including 135 anuran species. We tested the hypotheses that a predator's propensity to consume anuran young depends on (1) prey family, (2) predator manipulation strategy, and (3) prey ontogenetic stage. We used a binomial mixed model approach with considerations for multiple effect sizes within studies to evaluate the log odds ratio of the proportion of prey consumed by individual predators. Prey consumption was highly variable, but toads (Bufonidae) were consumed in lower proportions. Chewing invertebrates consumed more anuran prey than biting vertebrates. Late stage tadpoles were more vulnerable to predation than other stages of anuran ontogeny. However, more studies are needed to unravel the roles of development and evolutionary history in the chemical ecology of anuran young. This synthesis provides clear meta-analytic evidence that relative unpalatability is an important component in the anti-predator defenses of young in some anuran families, calling into question the degree to which chemically defended anuran families undergo ontogenetic switches in anti-predator strategies.

Dose-dependent alkaloid sequestration and N-methylation of decahydroquinoline in poison frogs.

Sequestration of chemical defenses from dietary sources is dependent on the availability of compounds in the environment and the mechanism of sequestration. Previous experiments have shown that sequestration efficiency varies among alkaloids in poison frogs, but little is known about the underlying mechanism. The aim of this study was to quantify the extent to which alkaloid sequestration and modification are dependent on alkaloid availability and/or sequestration mechanism. To do this, we administered different doses of histrionicotoxin (HTX) 235A and decahydroquinoline (DHQ) to captive-bred Adelphobates galactonotus and measured alkaloid quantity in muscle, kidney, liver, and feces. HTX 235A and DHQ were detected in all organs, whereas only DHQ was present in trace amounts in feces. For both liver and skin, the quantity of alkaloid accumulated increased at higher doses for both alkaloids. Accumulation efficiency in the skin increased at higher doses for HTX 235A but remained constant for DHQ. In contrast, the efficiency of HTX 235A accumulation in the liver was inversely related to dose and a similar, albeit statistically nonsignificant, pattern was observed for DHQ. We identified and quantified the N-methylation of DHQ in A. galactonotus, which represents a previously unknown example of alkaloid modification in poison frogs. Our study suggests that variation in alkaloid composition among individuals and species can result from differences in sequestration efficiency related to the type and amount of alkaloids available in the environment.

Dose-dependent alkaloid sequestration and N-methylation of decahydroquinoline in poison frogs

Sequestration of chemical defenses from dietary sources is dependent on the availability of compounds in the environment and the mechanism of sequestration. Previous experiments have shown that sequestration efficiency varies among alkaloids in poison frogs, but little is known about the underlying mechanism. The aim of this study was to quantify the extent to which alkaloid sequestration and modification are dependent on alkaloid availability and/or sequestration mechanism. To do this, we administered different doses of histrionicotoxin (HTX) 235A and decahydroquinoline (DHQ) to captive-bred Adelphobates galactonotus and measured alkaloid quantity in muscle, kidney, liver, and feces. HTX 235A and DHQ were detected in all organs, whereas only DHQ was present in trace amounts in feces. For both liver and skin, the quantity of alkaloid accumulated increased at higher doses for both alkaloids. Accumulation efficiency in the skin increased at higher doses for HTX 235A but remained constant for DHQ. In contrast, the efficiency of HTX 235A accumulation in the liver was inversely related to dose and a similar, albeit statistically nonsignificant, pattern was observed for DHQ. We identified and quantified the N-methylation of DHQ in A. galactonotus, which represents a previously unknown example of alkaloid modification in poison frogs. Our study suggests that variation in alkaloid composition among individuals and species can result from differences in sequestration efficiency related to the type and amount of alkaloids available in the environment.

Diverse traits of aquatic plants cannot individually explain their consumption by the generalist gastropod Biomphalairia glabrata.

Several experimental studies on aquatic plants have reported the prevalence of chemical defense mechanism against herbivory, as opposed to structural, life-forms or other traits. Here, our laboratory feeding experiments and integrative analysis explored the relationship among palatability (fresh or reconstituted plants used as artificial diet) and various chemical/nutritional traits (i.e., contents of dry mass, ash, nitrogen, protein, and phenols) of diverse aquatic plants and their susceptibility to consumption by the generalist gastropod Biomphalaria glabrata. Biomphalaria glabrata consumed all of the assayed aquatic plants in a hierarchical yet generalized way, with the consumption of fresh plants, their reconstituted forms and defensive properties of lipophilic extracts not being significantly correlated with plant physical or chemical traits to determine the feeding preference of the gastropod. Our results do not reveal a prevalence for a specific plant attribute contributing to herbivory. Instead, they indicate that the susceptibility of aquatic plants to generalist consumers is probably related to a combination of their chemical and physical properties, resulting in moderate grazing rates by generalist consumers.

Simulation of the electron ionization mass spectra of the Novichok nerve agent.

Novichok is one of the most feared and controversial nerve agents, which existence was confirmed only after the Salisbury attack in 2018. A new attack on August 2020, in Russia, was confirmed. After the 2018 attack, the agent was included in the list of the most dangerous chemicals of the Chemical Weapons Convention (CWC). However, information related to its electron ionization mass spectrometry (EI/MS), essential for unambiguous identification, is scarce. Therefore, investigations about Novichok EI/MS are urgent. In this work, we employed Born-Oppenheimer molecular dynamics through the Quantum Chemistry Electron Ionization Mass Spectrometry (QCEIMS) method to simulate and rationalize the EI/MS spectra and fragmentation pathways of 32 Novichok molecules recently incorporated into the CWC. The comparison of additional simulations with the measured EI spectrum of another Novichok analog is very favorable. A general scheme of the fragmentation pathways derived from simulation results was presented. The present results will be useful for elucidation and prediction of the EI spectra and fragmentation pathways of the dangerous Novichok nerve agent.

Domestication of Plants of Ugni molinae Turcz (Myrtaceae) Interferes in the Biology of Chilesia rudis (Lepidoptera: Erebidae) Larvae.

In terms of the domestication process in murtilla, studies have found changes in the concentration of phenolic compounds, with reduction of chemical defense of plants, depending on the change in the feeding behavior of insects. Thus, we hypothesized that the domestication of Ugni molinae decreases the content of phenolic compounds and modifies the feeding preference of Chilesia rudis larvae. Leaves of three parental ecotypes and four cultivated ecotypes were used in preference experiments to evaluate the mass gain and leaves consumption of larvae. Phenolic extracts from leaves of U. molinae were analyzed by HPLC. Identified compounds were incorporated in an artificial diet to assess their effect on mass gain, consumption, and survival of the larvae. The presence of phenolic compounds in bodies and feces was also evaluated. In terms of choice assays, larvae preferred parental ecotypes. Regarding compounds, vanillin was the most varied between the ecotypes in leaves. However, plant domestication did not show a reduction in phenolic compound concentration of the ecotypes studied. Furthermore, there was no clear relation between phenolic compounds and the performance of C. rudis larvae. Whether this was because of sequestration of some compounds by larvae is unknown. Finally, results of this study could also suggest that studied phenolic compounds have no role in the C. rudis larvae resistance in this stage of murtilla domestication process.

Effect of Spatial Variation on Defensive Substances of Constrictotermes Cyphergaster Soldiers (Blattaria, Isoptera).

The composition of chemical weaponry of termite soldiers show interspecific and intraspecific variation. However, spatial effects on the qualitative and quantitative compositions of these substances in Neotropical termites are poorly known. Hexane extracts of heads and the defensive secretion of soldiers of Constrictotermes cyphergaster from four localities in Northeast Brazil were analyzed by gas chromatography and mass spectrometry. Chemical analysis allowed the detection of 54 compounds from the head extract and from the direct extraction of the defensive secretion of soldiers, and the percentage of common substances and the presence of exclusive substances varied depending on the spatial distance between colonies. The profile of the chemical armament of soldiers consists basically of terpenoids: monoterpenes (45.53%-71.97 - for head extract and 57.41% - 78.56 for secretion) and sesquiterpenes (19.69% - 35.78% for head extract and 18.41% - 33.31%for secretion). In general, the main component of the chemical arsenal, regardless of the methodology used for extraction, was α-pinene (27.98-50.44%). Two chemotypes were identified based on chemical differences between populations of ecoregions with distinct spatial-environmental and climate characteristics: (1) α-pinene <33%; (2) α-pinene >33% for both extracts (head and secretion). The results reveal a similar pattern of chemical differentiation for soldiers in both extracts (head and secretion), with increasing differences as a function of distance between the analyzed colonies.

Review about Structure and Evaluation of Reactivators of Acetylcholinesterase Inhibited with Neurotoxic Organophosphorus Compounds.

BACKGROUND: Neurotoxic chemical warfare agents can be classified as some of the most dangerous chemicals for humanity. The most effective of those agents are the Organophosphates (OPs) capable of restricting the enzyme Acetylcholinesterase (AChE), which in turn, controls the nerve impulse transmission. When AChE is inhibited by OPs, its reactivation can be usually performed through cationic oximes. However, until today, it has not been developed one universal defense agent, with complete effective reactivation activity for AChE inhibited by any of the many types of existing neurotoxic OPs. For this reason, before treating people intoxicated by an OP, it is necessary to determine the neurotoxic compound that was used for contamination, in order to select the most effective oxime. Unfortunately, this task usually requires a relatively long time, raising the possibility of death. Cationic oximes also display a limited capacity of permeating the Blood-Brain Barrier (BBB). This fact compromises their capacity to reactivating AChE inside the nervous system. METHODS: We performed a comprehensive search on the data about OPs available on the scientific literature today in order to cover all the main drawbacks still faced in the research for the development of effective antidotes against those compounds. RESULTS: Therefore, this review about neurotoxic OPs and the reactivation of AChE, provides insights for the new agents' development. The most expected defense agent is a molecule without toxicity and effective to reactivate AChE inhibited by all neurotoxic OPs. CONCLUSION: To develop these new agents, the application of diverse scientific areas of research, especially theoretical procedures as computational science (computer simulation, docking and dynamics), organic synthesis, spectroscopic methodologies, biology, biochemical and biophysical information, medicinal chemistry, pharmacology and toxicology, is necessary.

The sea-hare Aplysia brasiliana promotes induction in chemical defense in the seaweed Laurencia dendroidea and in their congeneric neighbors.

Inducible chemical defenses are more common in temperate seaweeds than tropical ones, and are directly detected by increase of chemical contents, or indirectly by differential consumption of live seaweed tissues or artificial food with algal extracts by herbivores. In general, seaweed-induced chemical defense occur between 11 and 20 days after both simulated/artificial or direct herbivory. Here, we used experimental procedures to assess induced chemical defense in the tropical red seaweed Laurencia dendroidea as response to direct grazing, chemical cues from grazed conspecific neighbors and only presence of herbivores. Chemical defenses were analyzed by detecting the palatability of artificial food containing L. dendroidea extracts offered to Aplysia brasiliana and by comparative analyses of extracts from this seaweed by Gas Chromatography/Mass Spectroscopy, as well as metabolomic data analysis by Principal Component Analysis. Our results revealed that direct grazing by A. brasiliana induced a rapid (after 48 h) response among individuals of L. dendroidea, as did waterborne chemical cues from grazed conspecifics, but the presence of sea hare alone did not elicit a response. Increased resistance to grazing was accompanied by significative changes in sesquiterpene metabolomic chemical profile, revealing that induced defense: may be more widespread among seaweeds, independent of latitude; can involve changes in other classes of substances besides phlorotannins or not only the increase in the content of a single compound; and may be a rapid and ecologically coherent response to consumers. In addition, the importance of incorporating the metabolomic approach when examining inducible chemical defense in seaweeds is also emphasized.

New Ecological Role of Seaweed Secondary Metabolites as Autotoxic and Allelopathic.

Allelopathy and autotoxicity are well-known biological processes in angiosperms but are very little explored or even unknown in seaweeds. In this study, extract and major pure compounds from two distinct populations of the red seaweed Laurencia dendroidea were investigated to evaluate the effect of autotoxicity through auto- and crossed experiments under laboratory conditions, using chlorophyll fluorescence imaging to measure inhibition of photosynthesis (ΦPSII) as a variable response. Individuals of L. dendroidea from Azeda beach were inhibited by their own extract (IC50 = 219 µg/ml) and the major compound elatol (IC50 = 87 µg/ml); both chemicals also inhibited this seaweed species from Forno beach (IC50 = 194 µg/ml for the extract and IC50 = 277 µg/ml for elatol). By contrast, the extract of L. dendroidea from Forno and its major compound obtusol showed no inhibitory effect in individuals of both populations; but obtusol was insoluble to be tested at higher concentrations, which could be active as observed for elatol. The Azeda population displayed higher susceptibility to the Azeda extract and to elatol, manifested on the first day, unlike Forno individuals, in which the effect was only detected on the second day; and inhibition of ΦPSII was more pronounced at apical than basal portions of the thalli of L. dendroidea. This first finding of seaweed autotoxicity and allelopathic effects revealed the potential of the chemistry of secondary metabolites for intra- and inter-populational interactions, and for structuring seaweed populations.

Use of whole-body cryosectioning and desorption electrospray ionization mass spectrometry imaging to visualize alkaloid distribution in poison frogs.

Ambient mass spectrometry is useful for analyzing compounds that would be affected by other chemical procedures. Poison frogs are known to sequester alkaloids from their diet, but the sequestration pathway is unknown. Here, we describe methods for whole-body cryosectioning of frogs and use desorption electrospray ionization mass spectrometry imaging (DESI-MSI) to map the orally administered alkaloid histrionicotoxin 235A in a whole-body section of the poison frog Dendrobates tinctorius. Our results show that whole-body cryosectioning coupled with histochemical staining and DESI-MSI is an effective technique to visualize alkaloid distribution and help elucidate the mechanisms involved in alkaloid sequestration in poison frogs.

Molecular dynamics simulation of the electron ionization mass spectrum of tabun.

Tabun (ethyl N,N-dimethylphosphoramidocyanidate), or GA, is a chemical warfare nerve agent produced during the World War II. The synthesis of its analogs is rather simple; thus, it is a significant threat. Furthermore, experiments with tabun and other nerve agents are greatly limited by the involved life risks and the severe restrictions imposed by the Chemical Weapons Convention. For these reasons, accurate theoretical assignment of fragmentation pathways can be especially important. In this work, we employ the Quantum Chemistry Electron Ionization Mass Spectra method, which combines molecular dynamics, quantum chemistry methods, and stochastic approaches, to accurately investigate the electron ionization/mass spectrometry (EI/MS) fragmentation spectrum and pathways of the tabun molecule. We found that different rearrangement reactions occur including a McLafferty involving the nitrile group. An essential and characteristic pathway for identification of tabun and analogs, a two-step fragmentation producing the m/z 70 ion, was confirmed. The present results will be also useful to predict EI/MS spectrum and fragmentation pathways of other members of the tabun family, namely, the O-alkyl/cycloalkyl N,N-dialkyl (methyl, ethyl, isopropyl, or propyl) phosphoramidocyanidates.

Towards the optimization of botanical insecticides research: Aedes aegypti larvicidal natural products in French Guiana.

Natural products have proven to be an immeasurable source of bioactive compounds. The exceptional biodiversity encountered in Amazonia, alongside a rich entomofauna and frequent interactions with various herbivores is the crucible of a promising chemodiversity. This prompted us to search for novel botanical insecticides in French Guiana. As this French overseas department faces severe issues linked to insects, notably the strong incidence of vector-borne infectious diseases, we decided to focus our research on products able to control the mosquito Aedes aegypti. We tested 452 extracts obtained from 85 species originating from 36 botanical families and collected in contrasted environments against an Ae. aegypti laboratory strain susceptible to all insecticides, and a natural population resistant to both pyrethroid and organophosphate insecticides collected in Cayenne for the most active of them. Eight species (Maytenus oblongata Reissek, Celastraceae; Costus erythrothyrsus Loes., Costaceae; Humiria balsamifera Aubl., Humiriaceae; Sextonia rubra (Mez) van der Werff, Lauraceae; Piper hispidum Sw., Piperaceae; Laetia procera (Poepp.) Eichl., Salicaceae; Matayba arborescens (Aubl.) Radlk., Sapindaceae; and Cupania scrobitulata Rich., Sapindaceae) led to extracts exhibiting more than 50% larval mortality after 48 h of exposition at 100 µg/mL against the natural population and were considered active. Selectivity and phytochemistry of these extracts were therefore investigated and discussed, and some active compounds highlighted. Multivariate analysis highlighted that solvents, plant tissues, plant family and location had a significant effect on mortality while light, available resources and vegetation type did not. Through this case study we highlighted that plant defensive chemistry mechanisms are crucial while searching for novel insecticidal products.

Mechanisms of Convergent Egg Provisioning in Poison Frogs.

Parental provisioning of offspring with physiological products (nursing) occurs in many animals, yet little is known about the neuroendocrine basis of nursing in non-mammalian species. Within amphibians, maternal provisioning has evolved multiple times, with mothers of some species feeding unfertilized eggs to their developing offspring until tadpoles complete metamorphosis [1-3]. We conducted field studies in Ecuador and Madagascar to ask whether convergence at the behavioral level provides similar benefits to offspring and relies on shared neural mechanisms in dendrobatid and mantellid poison frogs. At an ecological level, we found that nursing allows poison frogs to provide chemical defenses to their tadpoles in both species. At the neural level, nursing was associated with increased activity in the lateral septum and preoptic area, demonstrating recruitment of shared brain regions in the convergent evolution of nursing within frogs and across vertebrates [4]. In contrast, only mantellids showed increased oxytocin neuron activity akin to that in nursing mammals [5], suggesting evolutionary versatility in molecular mechanisms. Our findings demonstrate that maternal provisioning provides similar potential benefits to offspring and relies on similar brain regions in poison frog species with convergently evolved toxicity and maternal care. VIDEO ABSTRACT.

Worker Defensive Behavior Associated with Toxins in the Neotropical Termite Neocapritermes braziliensis (Blattaria, Isoptera, Termitidae, Termitinae).

Termite societies are abundant in the tropics, and are therefore exposed to multiple enemies and predators, especially during foraging activity. Soldiers constitute a specialized defensive caste, although workers also participate in this process, and even display suicidal behavior, which is the case with the species Neocapritermes braziliensis. Here we describe the morphology, mechanisms of action, and proteomics of the salivary weapon in workers of this species, which due to the autothysis of the salivary glands causes their body rupture, in turn releasing a defensive secretion, observed during aggressiveness bioassays. Salivary glands are paired, composed of two translucent reservoirs, ducts and a set of multicellular acini. Histological and ultrastructural techniques showed that acini are composed of two types of central cells, and small parietal cells located in the acinar periphery. Type I central cells were abundant and filled with a large amount of secretion, while type II central cells were scarce and presented smaller secretion. Parietal cells were often paired and devoid of secretion. The gel-free proteomic approach (shotgun) followed by mass spectrometry revealed 235 proteins in the defensive secretion, which were classified into functional groups: (i) toxins and defensins, (ii) folding/conformation and post-translational modifications, (iii) salivary gland detoxification, (iv) housekeeping proteins and (v) uncharacterized and hypothetical proteins. We highlight the occurrence of neurotoxins previously identified in arachnid venoms, which are novelties for termite biology, and contribute to the knowledge regarding the defense strategies developed by termite species from the Neotropical region.

Testing for latitudinal gradients in defense at the macroevolutionary scale.

Plant defenses against herbivores are predicted to evolve to be greater in warmer climates, such as lower latitudes where herbivore pressure is also thought to be higher. Instead, recent findings are often inconsistent with this expectation, suggesting alternative hypotheses are needed. We tested for latitudinal gradients in plant defense evolution at the macroevolutionary scale by characterizing plant chemical defenses across 80 species of the evening primroses, spanning both North and South America. We quantified phenolics in leaves, flowers, and fruits, using advanced analytical chemistry techniques. Dominant individual ellagitannin compounds, total concentrations of ellagitannins, flavonoids, total phenolics, and compound diversity were quantified. Variation in these compounds was predicted with latitude, temperature, precipitation, and continent using phylogenetic generalized least squares (PGLS) multiple regression models. Latitude did not strongly explain variation in chemical defenses. Instead, fruit total ellagitannins, oenothein A, and total phenolics were greater in species inhabiting regions with colder climates. Using analytical chemistry and 80 species in two continents, we show that contrary to classic predictions, concentrations of secondary metabolites are not greater at lower latitudes or in warmer regions. We propose higher herbivore pressure in colder climates and gradients in resource availability as potential drivers of the observed patterns in Oenothera.

Behavior of uncharged oximes compared to HI6 and 2-PAM in the human AChE-tabun conjugate: a molecular modeling approach.

Tabun is one of the most dangerous nerve agents because it has deleterious effects like inhibition of the essential enzymes acetylcholinesterase (AChE) and butyrylcholinesterase. Some oximes such HI6 as 2-PAM are nucleophiles that are capable to reactivate inhibited human AChE under some conditions. Zwitterionic and cationic species have the best chance of productive action on inhibited AChE. However uncharged oximes can give important interaction information. In order to investigate the interaction and behavior of cationic and uncharged oximes, we performed molecular docking simulations and molecular dynamics and calculated binding energies of complexes of these compounds with human AChE. The uncharged oximes of larger structure were more susceptible to the influence of the substituents on the phosphorus atom and presented low binding energies. In contrast, HI 6 and 2-PAM showed high binding energy values with great contribution of the amino acid Asp74, demonstrating the importance of the quaternary nitrogen to the affinity and interaction of the oximes/AChE tabun-inhibited complexes.