Low Intraspecific Aggression Level, Cuticular Hydrocarbons, and Polydomy in the Bullet Ant.

Ants use chemical cues known as cuticular hydrocarbons (CHCs) for both intraspecific and interspecific recognition. These compounds serve ants in distinguishing between nestmates and non-nestmates, enabling them to coexist in polydomous colonies characterized by socially connected yet spatially separated nests. Hence, the aim of this study was to investigate the intraspecific aggression level between nestmates and non-nestmates of the bullet ant Paraponera clavata (Fabricius, 1775), analyze and compare their CHCs, and evaluate the occurrence of polydomy in this species. We conducted aggression tests between foragers, both in laboratory and field settings. To identify the chemical profiles, we utilized gas chromatography coupled with mass spectrometry (GC-MS). We marked the foragers found at nest entrances and subsequently recaptured these marked ants to validate workers exchange among nests. Across all nests, a low intraspecific aggression level was observed within the same area. However, a significant difference in aggression correlated to distance between nests. Analysis of the cuticular chemical profile of P. clavata unveiled colony-specific CHCs, both qualitatively and quantitatively. Notably, we observed instances of ants from certain nests entering or exiting different nests. This behavior, in conjunction with the observed low intraspecific aggression despite differences in CHCs suggests polydomy for this species. Polydomy can offer several benefits, including risk spreading, efficient exploitation of resources, potential for colony size increasing and reduced costs associated with foraging and competition.

Hydrocarbons in Formicidae: influence of chemical footprints on ant behavioral strategies.

When an insect walks, it leaves chemical cues that derive from the arolium, a tarsal structure. These cues may contain important information about other species that occur in their community and can then mediate interactions of competition, predation, and information about resources with ants from their own colony. The compounds of these cues are released into the substrate in the form of chemical footprints. There are still few species studied, and little is known about the behavior of ants regarding these signals and how they use them in their interactions. Therefore, the aim of this study was to assess the behavioral strategy of different ant species when confronted with chemical footprints left by other ants, as well as identify their compounds and their relationship with the cuticular hydrocarbon profile. The experiments were performed using a Y-maze, where in one of the arms, there were chemical footprints of their own species or of other species, and the other Y arm was footprint-free. The chemical compounds of footprints and cuticle were analyzed by gas chromatography-mass spectrometry. The results show that foragers of all species detect and respond to the presence of chemical cues in the form of footprints left by other ants. Foragers of all species followed footprints of individuals of the same species both nestmates and non-nestmates; however, Neoponera villosa avoided the footprints of Cephalotes borgmeieri, and C. borgmeieri avoided the footprints of the other two species. The chemical compositions of the cuticle and footprints are related to each other and are specific to each species.

Impacts of the insecticide thiamethoxam on the native stingless bee Plebeia catamarcensis (Hymenoptera, Apidae, Meliponini).

Agricultural production and the indiscriminate use of insecticides such as thiamethoxam have put at risk the biodiversity and ecosystem services provided by bees, including native stingless species. Since most of the native species do not present economic importance, they may suffer "silent extinction", due to lack of monitoring of their colonies. Therefore, this study aimed to determine the lethal and sublethal concentrations of the insecticide thiamethoxam, with evaluation of its sublethal effects on mobility, in the stingless bee Plebeia catamarcensis (Holmberg, 1903). Foraging bees were collected and exposed to thiamethoxam to determine lethal (LC50) and sublethal concentrations. The 24 h LC50 was 0.408 ng a.i./µL, a value demonstrating that this species may be as sensitive as other stingless bees already studied. Sublethal concentrations influenced the locomotion abilities of the bees, making them hyperactive when exposed to LC50/10 and lethargic when exposed to LC50/100. The effects of sublethal concentrations on individuals may have collective consequences, especially in colonies with few individuals, as is the case of P. catamarcensis. The findings reinforce the hypothesis that thiamethoxam may contribute to the decline of native stingless bees, which can be significantly impacted when chronically exposed to agricultural production systems that use this insecticide, consequently affecting the ecosystem services provided by these bees.

Effects of chronic exposure to sublethal doses of neonicotinoids in the social wasp Polybia paulista: Survival, mobility, and histopathology.

Several studies have investigated the consequences of exposure to neonicotinoids in honeybees. Given the lack of studies concerning the consequences of exposure of social wasps to neonicotinoids, as well as the ecological importance of these insects, the objective of this study was to test the hypothesis that chronic exposure to sublethal concentrations of thiamethoxam decreases survival and mobility by causing damage to the brain and midgut of the social wasp Polybia paulista. The wasps were exposed to different concentrations of thiamethoxam, in order to obtain the mean lethal concentration (LC50), which was used as a reference for calculation of two sublethal concentrations (LC50/100 and LC50/10) employed in subsequent experiments. To calculate survival, groups of exposed (EW) and unexposed (UW) wasps were monitored until death, allowing calculation of the average lethal time. The EW and UW groups were evaluated after 12, 24, 48, and 72 h of exposure, considering their mobility and histopathological parameters of the midgut and brain. A lesion index based on semiquantitative analyses was used for comparison of histopathological damage. The results demonstrated that exposure to the LC50/10 led to a significantly shorter survival time of the P. paulista workers, compared to unexposed wasps. In addition, both sublethal concentrations decreased mobility and caused damage to the intestine (loss of brush border, presence of spherocrystals, loss of cytoplasmic material, and pyknosis) and the brain (loss of cell contact and pyknosis), regardless of the exposure time. The findings showed that, like bees, social wasps are nontarget insects susceptible to the detrimental consequences of neonicotinoid use, with exposure leading to impaired survival, locomotion, and physiology.

Lethal and Sublethal Dose of Thiamethoxam and Its Effects on the Behavior of a Non-target Social Wasp.

In the last few years, with the increase in agricultural productivity, there has also been an increase in the use of insecticides to combat insects considered pests. However, these chemical compounds end up affecting nontarget insects that also interact with the crops. Studies have shown that social bees are among the insects that are suffering most from the effects of these compounds, resulting in negative ecological and economic impacts, considering that these insects provide pollination services in ecosystems. At the same time, social wasps also interact with plants, including cultivated ones, and perform ecological services similar to those of social bees, so it can be hypothesized that insecticides are also affecting social wasp colonies. Therefore, the purpose of this study was to evaluate contamination and sublethal effects of neonicotinoids on the mobility of the social wasp Protopolybia exigua (Saussure). In the first step, oral exposure experiments were performed to determine lethal and sublethal concentrations. In a second step, the wasps were exposed to sublethal concentrations, in order to evaluate the effects on their mobility. The results demonstrated that this species is more susceptible to exposure to neonicotinoids, compared to several bee species that have so far been studied, but lower than others. Exposure to sublethal concentrations can significantly reduce wasp mobility, which can have short-term consequences both for worker wasps and for the maintenance of their colonies.

Magnetic nanoparticles in the body parts of Polistes versicolor and Polybia paulista wasps are biomineralized: evidence from magnetization measurements and ferromagnetic resonance spectroscopy.

The detection of the geomagnetic field by animals to use as a cue in homing and migration is known as magnetoreception. The ferromagnetic hypothesis explains magnetoreception assuming that magnetic nanoparticles in cellular structures are used as magnetic field transducers. Considering magnetoreception in social insects, the most studied has been the honeybee Apis mellifera and only in two wasp species (Vespa orientalis and Polybia paulista) have been shown a magnetosensitive behavior. In the present report the body parts (abdomen, head and antennae) of Polistes versicolor and Polybia paulista wasps were studied aiming to find biomineralized magnetic nanoparticles, using magnetometry measurements and ferromagnetic resonance spectroscopy. The magnetometry measurements show the presence of magnetic nanoparticles in all body parts, being characterized as mixtures of superparamagnetic, single domain and pseudo-single domain nanoparticles. From the ferromagnetic resonance spectra were obtained the asymmetry ratio A and the effective g factor geff, and those parameters are consistent with the presence of biomineralized magnetic nanoparticles in both wasps. In the case of Polybia paulista, the magnetic nanoparticles can be associated with some sort of magnetosensor once this wasp is magnetosensitive. For Polistes versicolor, the results indicate that this wasp can be magnetosensitive as Polybia paulista once their magnetic nanoparticles are biomineralized in the body. Behavioral studies with Polistes versicolor wasps deserve to be performed.

Effect of thiamethoxam (organophosphate) on the flies and beetle visitation and cadaveric decomposition process

ABSTRACT Entomofauna associated with decaying cadavers may be useful in criminal investigation, either through the development of immature insects of interest or through entomological succession in corpses. These factors may vary if the insects are exposed to toxic substances that can modify the insect's developmental cycle, as well as its own occurrence, which would imply significant changes in the results of any investigation. However, there are few studies on how contamination by toxic compounds can affect the action of insects on carcasses and their consequence for forensic expertise. Therefore, this study aimed to test the hypothesis that the normal visitation of flies and beetles is altered in insecticide-contaminated carcasses. The experiment was carried out in a sugarcane plantation, using pig carcasses contaminated with insecticide and the same number of carcasses without any type of contamination as a control. In all experiments, the contaminated carcasses reached the final phase of decomposition in a longer time than the uncontaminated carcasses of the control group. A total of 2.767 specimens were collected and identified, 2.103 individuals from the order Diptera and 664 from the order Coleoptera. There was a significant geometric regression adjustment during the decomposition phases only for the control group, different from the contaminated carcasses in which this pattern did not occur. Results show that contamination by insecticide alters decomposition time and phase, altering the action of flies and beetles, affecting the abundance, composition of species as well as their activities, which can alter the data used by experts in criminal experts.

Is the social wasp Polybia paulista a silent victim of neonicotinoid contamination?

Neonicotinoids are among the chemicals most widely used against insects considered agricultural pests, although they may also affect nontarget species, as has been reported for social bees. Social wasps are recognized as efficient predators of larvae of other insects, including pest species, so they may have contact with insecticides, at least indirectly. However, to date, there have been no studies investigating the consequences for social wasps of the use of neonicotinoids. Therefore, the aim of this study was to obtain the topical and oral acute lethal mean doses of the neonicotinoid thiamethoxam, as well as to evaluate the effects of sublethal contamination, for the social wasp Polybia paulista. Foraging wasps were exposed orally and topically, with monitoring of their mortality after 24 and 48 h, in order to calculate the LD50 values. Other wasps were then exposed with sublethal doses, in order to evaluate the effects of contamination on their mobility. The results indicated that P. paulista workers are as sensitive to thiamethoxam contamination as several nontarget species studied previously. Exposure to a sublethal dose (LD10) negatively affected their mobility. It could be concluded that social wasp colonies may be as much affected by exposure to neonicotinoids as social bees. Further toxicological studies should be conducted, evaluating other parameters and different species, in order to understand the extent of the problems faced by these insects due to the use of neonicotinoids.

Is It Possible to Obtain the Chemical Profile From Ethanol-Preserved Specimens? The Hydrocarbon and Fatty Acid Composition of the Social Wasp Polybia paulista (Hymenoptera: Vespidae: Epiponini).

Cuticular hydrocarbons perform multiple functions in insects such as protecting against desiccation and pathogenic infection, and signaling interactions. Evaluation of cuticular hydrocarbon (CHC) profiles of insects is commonly performed by extraction using a nonpolar solvent such as hexane. Specimens intended for CHC analysis are ideally handled by avoiding contact with solvents such as ethanol. However, insects are frequently stored in ethanol after collection, especially if intended for molecular analysis. To determine if it is possible that chemical compounds in the cuticles of specimens can withstand previous exposure to ethanol, we evaluated the efficiency of CHC extraction from specimens preserved in 95% ethanol. We extracted cuticular compounds from specimens of the social wasp Polybia paulista (Ihering) with no contact with ethanol solvents and compared them with those from specimens stored in 95% ethanol. We analyzed chemical composition from wasps and the 95% ethanol in which they had been stored by a gas chromatograph coupled to a mass spectrometer. In total, 56 compounds were detected: 50 that were classified as hydrocarbons which were mostly branched alkanes, followed by linear alkanes and alkenes. Three compounds were identified as fatty acids, and three compounds were unidentifiable. The ethanol-preserved specimens showed similar chemical profiles to those of specimens that had no contact with ethanol. Thus, we suggest that it is possible to study the chemical profiles of ethanol-preserved specimens.

A 3D model to illustrate the nest architecture of Acromyrmex balzani (Hymenoptera; Formicidae)

ABSTRACT For eusocial insects, the nest is a place where the main social interactions occur. The nest architecture ensures protection from predators and the environment, as well as suitable conditions for brood rearing, food storage, and in some cases the cultivation of fungus farms. Variations in nest architecture can occur, according to the environmental conditions. In order to elucidate the internal organization of nests, most studies use 2D schemes and photographs to illustrate the nest architecture models. However, 3D models can provide a different and more realistic view of the nest architecture. The aim of this study was to describe the nest architecture and colony size of the grass-cutting ant Acromyrmex balzani (Emery), using 3D models to illustrate these features. The structures of eight colonies were measured and the data were used to create a 3D model of each nest. Externally, the nests had one or more piles of loose soil and waste, with a single straw turret over the entrance. Underground, the nests had from 2 to 6 chambers, at a maximum depth of 122 cm. It could be concluded that the observed nest architecture of Acromyrmex balzani followed, at least in part, the pattern already reported in the literature. However, this is the first report of connection between two chambers made by two shafts, as well as the presence of the turret at the nest entrance/exit, regardless of the season of the year. These differences evidence that the nest structures may vary, depending on intrinsic or local environmental conditions.

New approach to application of mid-infrared photoacoustic spectroscopy in forensic analysis: Study with the necrophagous blow fly Chrysomya megacephala (Diptera: Calliphoridae).

Insects can provide clues in a variety of ways to assist in criminal investigations. The FTIR-PAS technique has been successfully used to assess the cuticular chemical profiles of insect samples from different groups and for several goals. However, until now, it has never been used to evaluate samples of forensic interest, despite providing faster results, compared to the methods currently used. In this study, mid-infrared photoacoustic spectroscopy was employed to assess the cuticular chemical profiles of different stages of development of the blow fly Chrysomya megacephala sampled from two distinct populations. The results showed that this technique enabled detection of significant differences between the main vibrational modes of the chemical bonds present in the cuticles of the two populations and the different stages of development of the fly. The method enables identification of the age of individuals collected at the crime scene, as well as the distinction of different populations. Therefore, this methodology could assist in forensic investigations, in both estimating the Postmortem Interval and determining the location where the crime occurred, or whether the body had suffered some type of translocation. The technique provides high reproducibility and fast analysis, so its application for analysis of C. megacephala is a viable option in forensic crime investigations.

Proteomic analysis of the venom of the social wasp Apoica pallens (Hymenoptera: Vespidae)

ABSTRACT Wasps are a diverse group of insects that possess a sting apparatus associated with a venom gland, which is used for predation and colony defense. The biochemistry of Hymenoptera venom has been evaluated in relation to allergy and immunology, and proteomics has been shown to be a powerful tool for the identification of compounds with pharmacological potential. Data on wasps venom the of genus Apoica are scarce, so the objective of the present work was to identify the venom proteins of the eusocial wasp Apoica pallens, as a first step towards further investigation of applied uses of the venom and its protein constituents. The venom proteins were separated by two-dimensional gel electrophoresis, followed by MALDI-TOF/TOF mass spectrometry. A total of 259 spots were detected, with molecular weights from 4.9 to 141 kDa. Thirty of these proteins were identified and classified into eight functional categories: allergen, enzyme, metabolism, structural, environmental response, proteoglycan, active in DNA and RNA, and unknown function. Due to the few available proteomic data for wasp venom, many proteins could not be identified, which makes studies with proteomic analysis of Hymenoptera venom even more important.

Can altered magnetic field affect the foraging behaviour of ants?

Social insects such as ants can use geomagnetic field information in orientation and navigation tasks. However, few studies have assessed the effect of magnetic fields on aspects such as orientation and decision making during foraging of ants. Therefore, the present study aims to test the hypothesis that foragers of different species of ants with different foraging strategies when under effect of applied magnetic field change the patterns of search for resources and recruitment of ants. We used two species with solitary foraging strategy, Ectatomma brunneum and Neoponera inversa, and another with mass recruitment, Pheidole sp. The experiments were performed in field and laboratory conditions. We used some parameters for comparison such as speed, distance and time during foraging in the field and laboratory experiments, under normal and applied magnetic field with the coils on and off. We also performed SQUID magnetometry analysis for all species. The results demonstrate that changes in normal values of magnetic field affect workers behaviour of the three species. Thus, we can conclude that ants under the effect of applied magnetic fields can suffer significant changes in their foraging activities decreasing the flow of workers, increasing the travelled distance from the nest to the resource and back to the nest, in addition to time and distance to fetch the resource and decision-making, in both types of species, those which have mass recruitment, or forage individually, and that the three species are magnetosensitive, being affected by changes of low intensity in the local magnetic field.

Hydrocarbon and Fatty Acid Composition from Blowfly Eggs Represents a Potential Complementary Taxonomic Tool of Forensic Importance.

One of the most important contributions of forensic entomology is to assist criminal expertise to determine the postmortem interval, which depends on the duration of the immature stages of insects of forensic interest. On the other hand, the time of development of the different stages varies according to the species; therefore, its identification is essential. Currently, few studies have investigated the use of cuticular hydrocarbons, and none regarding fatty acids, as complementary taxonomic tools to expedite species identification. Therefore, we evaluated whether cuticular hydrocarbons together with fatty acids of eggs of flies of the family Calliphoridae, main group of forensic interest, can be used to distinguish species. The analyses were performed by chromatographic techniques. The results show that there are significant differences between the composition of cuticular hydrocarbons and fatty acids between species and, therefore, they can be used to provide a complementary taxonomic tool to expedite the forensic expertise.

Effect of temperature on the chemical profiles of nest materials of social wasps.

Social insects depend on their nests for protection against predation and abiotic threats. Accordingly, the chemical compounds present in the material wasps use to build their nests can both facilitate communication and repel predators. It is herein hypothesized that different wasp species build their nests with different structure and substrate materials and that such materials consist of chemical compounds related to unique wasp behavior and outside temperature variation. To test this hypothesis, nests were collected from three species of social wasps, the samples of which were subjected to temperature variation under laboratory conditions. The compounds present in the substrate were analyzed by gas chromatography coupled to mass spectrometry. Chemical compounds identified in the nest material of the three species responded differently to temperature variation. Chemical compounds from Polybia nests were altered significantly when subjected to temperature variation, whereas the nests of Polistes versicolor did not significantly change in relation to the control. The differences found between Polistes and Polybia nests may be related to genetic factors, but also to the type of nest they construct. It is possible that divergent evolutionary strategies for maintaining colony temperature, as a function of the chemical composition of the nests, may have appeared between wasps that have open and closed nests. In relatively small colonies, nest substrate is more resistant to temperature variation because it is composed of a greater diversity of elements and thus capable of holding heavier, longer carbon chains. Our results suggest that chemical compounds in the nest material of the three wasp species analysed responded differently to fluctuating ambient temperatures and that such variation could result from the biochemical differences of unique wasp species or from thermoregulation strategies of colonies.

Effect of temperature on survival and cuticular composition of three different ant species.

Climatic factors, such as temperature variation, interfere with the survival of insects. To respond to these variations, insects have some specific characteristics. These include water content of the body, thickness of the lipid layer, as well as the qualitative and quantitative characteristics of cuticular chemical components. This study hypothesizes that different ant species respond to temperature changes in different ways and that such differences may be associated with cuticle hydrocarbons (CHCs) and fatty acids. As model ant species, Atta sexdens, Odontomachus bauri and Ectatomma brunneum were used for experimental analyses. Ants were submitted to a water bath for 5 h at different temperatures, and their CHCs and fatty acids were identified and quantified, followed by correlating these chemical compounds with temperature variations and the survival. Temperatures below 30 °C did not affect the survival of the three species. E. brunneum had a higher percentage of survival at temperatures above 30 °C. O. bauri was the most sensitive species with 100% mortality at 40 °C. Survival was found to be unrelated to any of the identified fatty acids. However, CHCs underwent significant quantitative and qualitative variation, as shown by an increased percentage of CHCs with longer chain length of linear alkanes at temperatures above 30 °C. These increase enables these ants to maintain the integrity of their cuticle and survive at temperatures above 30 °C. It can be concluded that the forager ants studied respond differently to temperature variation and that changes in the conformation of CHCs are in line with the ecological characteristics of the different studied species because, they vary in terms of diurnal/nocturnal foraging and types of environments foraged. Among the three species, E. brunneum foragers were found to be more active under adverse conditions and more tolerant to temperature variation with the correspondingly appropriate changes in CHCs composition.

New method for estimating the post-mortem interval using the chemical composition of different generations of empty puparia: Indoor cases.

Most flies of forensic importance are in two superfamilies, the Muscoidea and the Oestroidea, with similar life stages including the puparium. Upon completion of metamorphosis the adult fly emerges from the puparium, leaving behind an exuvia that is of potential significance in forensic investigation. The empty puparium is a durable piece of entomological evidence lasting several years. Through the study of chemical compounds, specifically the hydrocarbons of these puparia, it is possible to identify the species, in addition to how long they have been exposed to weathering and for this reason, these parameters can assist forensic entomologists in estimating long-term postmortem interval (minPMI). In corpses that take a relatively longer time to decompose, insects may use the same corpses for several oviposition cycles. Therefore, the aim of this study was to develop a new method to determine the PMI based on chemical compounds of the puparia from different oviposition cycles of the fly Chrysomya megacephala. The chemical composition of 50 puparia from different cycles of oviposition were evaluated by Gas Chromatography-Mass Spectrometry (GC-MS). In total, 60 compounds were identified ranging from C18 to C34, 38 of those were common to all generations. Our results demonstrate that chemical profiles can be used to differentiate puparia collected from successive cycles, and therefore valuable in the estimation of minPMI.

Influence of temperature on survival and cuticular chemical profile of social wasps.

The cuticle of social insects is a barrier against desiccation and a channel for chemical communication, two characteristics fundamental to the success of this group. The compounds present in the cuticle interact dynamically in order to achieve a balance between these two functions. Thus, viscosity correlates with waterproofing, whereas fluidity correlates with effective communication. Temperature variation can cause the cuticular hydrocarbon (CHC) layer of the cuticle to change in order to maintain body homeostasis. Thus, in this study, we tested the hypothesis that wasps with different body sizes and nest types will differ in their tolerance to temperature variation and ability to respond by changing cuticular chemical composition. To test this hypothesis, workers of three species of social wasps with different body sizes and nests, both with or without envelope, were subjected to different temperatures under controlled conditions. Cuticular compounds were analyzed by gas chromatography coupled to mass spectrometry (GC/MS). Results show that tolerance to temperature variation is not directly related to either wasp size or nesting type. An increase in the percentage of linear alkanes and a decrease in the percentage of branched alkanes were correlated with increased temperature. Thus, instead of either body size or nest type, tolerance to temperature variation seems to be mediated by the changing chemical composition of the cuticle.

Observation of intra- and interspecific differences in the nest chemical profiles of social wasps (Hymenoptera: Polistinae) using infrared photoacoustic spectroscopy.

The aim of this study was to explore whether the nest chemical profile (NCP) can be used to determine intra- and interspecific differences in social wasps of the subfamily Polistinae. For this purpose, Fourier transform infrared photoacoustic spectroscopy (FTIR-PAS) was used to directly analyze small pieces of nest as well as the gasters of females. An advantage of the methodology was that no sample preparation was required. FTIR-PAS combined with multivariate discriminant analyzes was used, to the best of our knowledge for the first time, to evaluate the NCPs of six species of polistine wasps, observing the influence of the nesting environment and investigating the relationships among the nest and female cuticular chemical profiles. The results revealed significant inter-species differences among the NCPs, as well as strong correlations with the environments in which the nests were located. The Mischocyttarus and Polistes species belong to the same tribe (Polistini) and therefore exhibited similar NCPs. These species were separated from the Polybia species in the NCP dendrogram, mainly because the Polybia belong to another tribe (Epiponini). Correlation was observed between the NCPs and the cuticular chemical profiles of females. The findings of the study demonstrated the importance of the NCP for differentiation of species and environments, and the utility of FTIR-PAS for identification of correlations between individuals and nests. The results confirmed the hypothesis that the NCP can be used as an additional tool for intra- and interspecific differentiation in social wasps of the subfamily Polistinae.

Intraspecific variation and influence of diet on the venom chemical profile of the Ectatomma brunneum Smith (Formicidae) ant evaluated by photoacoustic spectroscopy.

Studies of venomous animals have shown that environmental and genetic factors contribute to determining the chemical composition of venom. It is well known that external effects cause differences in the toxicity, concentration, and prey specificity of venom. However, the influence of different factors on the chemical profile of Hymenoptera venom remains little explored. In view of this, the aim of this study was to evaluate intraspecific differences and the influence of diet on the chemical profile of Ectatomma brunneum venom using Fourier transform infrared photoacoustic spectroscopy. For the study of intraspecific variation of the venom, foragers were collected at locations with different environmental conditions, such as urban, intermediate, woodland and monoculture sites. To investigate the influence of diet on the venom, two colonies were sampled in the same area and were maintained in the laboratory under controlled diet conditions and at room temperature. The mid-infrared absorption spectra obtained were interpreted using discriminant function analysis. The results showed significant differences among the chemical profiles of the venoms of individuals from different environments and individuals exposed to a controlled diet in the laboratory, suggesting that venom composition was determined not only by genetic traits inherent to the species, but also by exogenous factors.