Submitted by DIEGO FELISBINO FRAGA null (diegofraga@fazu.br) on 2016-03-08T21:00:47Z No. of bitstreams: 1 Tese_Diego_Felisbino_Fraga.pdf: 1816202 bytes, checksum: 645838702bb204f2d81827bcc74d32f0 (MD5) / Approved for entry into archive by Ana Paula Grisoto (grisotoana@reitoria.unesp.br) on 2016-03-09T18:05:33Z (GMT) No. of bitstreams: 1 fraga_df_dr_jabo.pdf: 1816202 bytes, checksum: 645838702bb204f2d81827bcc74d32f0 (MD5) / Made available in DSpace on 2016-03-09T18:05:33Z (GMT). No. of bitstreams: 1 fraga_df_dr_jabo.pdf: 1816202 bytes, checksum: 645838702bb204f2d81827bcc74d32f0 (MD5) Previous issue date: 2016-02-22 / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / O Controle Biológico Natural, ou Conservativo, é uma ferramenta indispensável para o sucesso do Manejo Integrado de Pragas, pois permite uma interação entre os componentes do ambiente, o que o coloca na linha de frente das táticas e estratégias de manejo de pragas em agroecossistemas. Basicamente, esta interação, denominada interação tritrófica, envolve as plantas, os organismos fitófagos e seus inimigos naturais. Neste contexto, diversos fatores são importantes para o controle biológico, tais como condições abióticas e bióticas, destacando-se características das plantas (compostos metabólitos secundários e arquitetura, por exemplo) e potenciais presas/hospedeiros (semioquímicos, etc.). Inimigos naturais podem utilizar estes recursos durante o seu processo de busca e localização do hospedeiro, de modo que o conhecimento sobre o comportamento de inimigos naturais em função da fenologia e arquitetura das plantas, bem como sobre o potencial de voláteis emitidos por plantas e presas é fundamental para um programa de manejo integrado de pragas. Desta forma, o presente trabalho teve por objetivos: a) avaliar a dinâmica populacional e a distribuição vertical de ovos de Chrysodeixis includens (Lepidoptera: Noctuidae) e de seus ovos parasitados por Trichogramma spp. (Hymenoptera: Trichogrammatidae) na cultura da soja; b) determinar a resposta comportamental do percevejo predador Orius insidiosus (Hemiptera: Anthocoridae) para voláteis associados à Halyomorpha halys (Hemiptera: Pentatomidae). Sendo assim, em Jaboticabal, SP, Brasil, durante as safras agrícolas de 2012/13 e 2013/14, foram realizadas coletas de ovos de C. includens presentes em diferentes terços das plantas de soja, durante diferentes estádios fenológicos da cultura. Semanalmente, o número de ovos presentes nas plantas foi anotado e os mesmos eram coletados e levados para laboratório para aguardar a emergência de lagartas ou de adultos de Trichogramma spp. O número de adultos do parasitoide emergido por ovo era registrado, bem como a razão sexual. Os adultos foram identificados ao nível específico, encontrando-se a espécie Trichogramma pretiosum presente na cultura da soja. Fêmeas de C. includens preferiram ovipositar com mais intensidade nos terços médio e inferior das plantas, principalmente durante os estádios reprodutivos da cultura. O parasitismo de ovos por T. pretiosum apresentou uma tendência semelhante, em que a medida que se aumentou a densidade de ovos de C. includens nas plantas, também elevou-se o número médio de ovos parasitados, indicando uma forte correlação positiva entre a praga e o inimigo natural. Para atingir o segundo objetivo desta tese, e fornecer alternativas para o controle biológico de H. halys, um pentatomídeo invasor nos EUA, foram conduzidos experimentos para testar a atratividade de O. insidiosus para voláteis emitidos pela presa e por plantas de feijoeiro atacadas por ela. Foram conduzidos experimentos em casa-de-vegetação, laboratório e em campo, em que foi identificado altas concentrações do composto tridecano. Foi avaliado a atratividade de O. insidiosus para diferentes concentrações deste composto, bem como os efeitos desta atração na predação de ovos. Foi observado que O. insidiosus é atraído por tridecano, no entanto, esta atração não afeta a predação de ovos, em casa de vegetação e no campo. / Natural ou Conservative Biological Control is a valuable tool for the success of Integrated Pest Management as it allows a natural interaction between the components of the environment, which places it at the forefront of the tactics and pest management strategies in agroecosystems. This interaction, so-called tritrophic interaction, involves plants, phytophagous organisms and their natural enemies. In this context several factors are important for biological control, such as biotic and abiotic conditions, highlighting plant characteristics (secondary metabolites compounds and plant architecture, for example) and possible prey/hosts (semiochemicals, etc.). Natural enemies can use this information during the its host foraging behavior, so the knowledge of the behavior of natural enemies according to the phenology and plant architecture as well as potential volatiles emitted by prey plants and potential is key for an integrated pest management program. Thus, this study aimed to: a) assess the population dynamics and vertical distribution of natural biological control of Chrysodeixis includens (Lepidoptera: Noctuidae) eggs by Trichogramma spp. (Hymenoptera: Trichogrammatidae) in soybean field; b) determine the behavioral response of the predator Orius insidiosus (Hemiptera: Anthocoridae) for volatile associated with Halyomorpha halys (Hemiptera: Pentatomidae). Thus, in Jaboticabal, SP, Brazil, during the growing seasons of 2012/2013 and 2013/2014, samples were taken from C. includens eggs present on different thirds of soybean plants during different phenological stages of the crop. Weekly, number of eggs present in plants was recorded and the eggs were collected ant taken to the laboratory to await the emergence of larvae or adult Trichogramma spp. The number of adult Trichogramma spp. emerged per egg was noted, as was the sex ratio. The adults were identified to species level, finding Trichogramma pretiosum as the main species present in soybeans. Females of C. includens intensively preferred to lay eggs in the middle and lower thirds of the plants, especially during the reproductive stages of soybean plants. Egg parasitism by T. pretiosum presented a similar tendency, which as the C. includens eggs density increased in plants, also increased the average number of parasitized eggs, indicating a strong positive correlation between the pest and the natural enemy. To achieve the second objective of this thesis, and provide alternatives for the biological control of H. halys (an invasive pentatomid in the US). experiments were conducted to test the attractiveness of O. insidiosus to volatiles emitted by the prey and bean plants attacked by it. Experiments were conducted under greenhouse, laboratory and field conditions. It was identified that tridecane was the major compound in damage-bean pods as well as on H. halys. The attractiveness of O. insidiosus was evaluated for different concentrations of this compound as well as the effects of this attraction as predators of eggs. It was observed that O. insidiosus is attracted to tridecane, however, this attraction not poisitivamente affected the predation of eggs in the greenhouse and in the field. / CNPq: 164869/2014-0

Submitted by JOSÉ PAULO GONÇALVES FRANCO DA SILVA null (jpgfdsilva@fca.unesp.br) on 2016-09-14T19:15:06Z No. of bitstreams: 1 Bentivenha JPF_Dissertation_2016.pdf: 2117731 bytes, checksum: 70ae67a18a3d561f32eacb362814d40d (MD5) / Approved for entry into archive by Ana Paula Grisoto (grisotoana@reitoria.unesp.br) on 2016-09-19T19:12:10Z (GMT) No. of bitstreams: 1 silva_jpgf_dr_bot.pdf: 2117731 bytes, checksum: 70ae67a18a3d561f32eacb362814d40d (MD5) / Made available in DSpace on 2016-09-19T19:12:10Z (GMT). No. of bitstreams: 1 silva_jpgf_dr_bot.pdf: 2117731 bytes, checksum: 70ae67a18a3d561f32eacb362814d40d (MD5) Previous issue date: 2016-08-19 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Entre as principais pragas da cultura do milho, destacam-se quatro espécies de lepidópteros da família Noctuidae: a lagarta-do-cartucho Spodoptera frugiperda (J.E. Smith), a lagarta-da-espiga Helicoverpa zea (Boddie), Helicoverpa armigera (Hübner), e Striacosta albicosta (Smith). Essas espécies podem interagir e competir por alimento, uma vez que apresentam a mesma guilda alimentar, a espiga do milho. Canibalismo/predação também são comportamentos também apresentados durante o estágio larval para a maioria dessas espécies, o que intensifica a competição intraguilda. Somado a isso, o tipo de agroecossistema, que oferece uma grande variação de plantas hospedeiras para as espécies-praga, a recente identificação de H. armigera no continente americano, e também a disperssão de S. albicosta nos EUA, tem trazido vários questionamentos sobre o impacto das interações larvais no manejo integrado de pragas (MIP) e no manejo de resistência dos insetos (MRI). Além disso, o complexo manejo dessas espécies, juntamente com a adoção de híbridos de milho transgênicos (Bt), as doses subletais de Bt, e as estratégias de manejo da resistência, envolvendo as estratégias de refúgio, eleva a importância do comportamento de noctuídeos. Compreender o comportamento e interações larvais, primeiramente em milho não-Bt, é essencial para entender a prevalência de espécies nas culturas agrícolas, e para a obtenção de sucesso no MIP e MRI. Sendo assim, esse estudo avaliou as interações intraguilda de lagartas de diferentes espécies, bem como seus movimentos, através de estudos comportamentais em milho não-Bt. Inicialmente, as interações intraguilda de S. frugiperda, H. zea e S. albicosta foi investigada em condições de laboratório e campo nos EUA, através de cenários de interação em arenas específicas e estruturas de milho. Em outro estudo em laboratório, foi avaliado os movimentos de ataque e defesa entre S. frugiperda e H. zea em diferentes cenários de interação, com e sem alimentação, constituindo um estudo de etograma. Um estudo entre as espécies, anteriormente alopátricas, H. zea e H. armigera, foi conduzido envolvendo a interação intraguilda na cultura do milho, em competição intraespecífica e interespecífica, assim como a ocorrência e proporção das espécies em três regiões brasileiras. Finalmente, a interação intraguilda de S. frugiperda com Helicoverpa spp. foi investigada em cenários em condições de campo de laboratório, em milho no Brasil. Um sistema de rastreamento de vídeo foi utilizado para caracterizar parâmetros de comportamento larval resultado das interações. Em relação ao primeiro estudo, S. frugiperda apresentou vantagem nas interações sobre as outras espécies, enquanto que S. albicosta apresentou desvantagem nas interações. No segundo estudo, S. frugiperda exibiu predominância de movimentos defensivos quando interagindo com H. zea em mesmo ínstar. Por outro lado, lagartas de H. zea apresentaram predominância de comportamentos agressivos, e com canibalismo/predação ocorrendo em interações de sexto ínstar como oponentes de quarto ínstar. No terceiro estudo, H. zea foi predominante no Rio Grande do Sul e na região central do Brasil. No oeste da Bahia, a espécie foi predominante na primeira coleta, sendo a relação H. zea: H. armigera similar na segunda coleta. Lagartas de H. zea apresentaram vantagem na interação com H. armigera. O agroecossistema tem importante papel na ocorrência das espécies e impacta na interação e prevalência destas no meio ambiente. Lagartas de S. frugiperda apresentam vantagens na competição com Helicoverpa spp. Frequência e tempo se alimentando de lagartas de S. frugiperda são negativamente afetadas pelas interações. Lagartas de S. frugiperda movem curtas distâncias em comparação com H. zea. / Among the insect pests from maize crop, stand out the lepidopteran species belonging to Noctuidae family, covering the fall armyworm Spodoptera frugiperda (J.E. Smith), the corn earworm Helicoverpa zea (Boddie), the Old World bollworm Helicoverpa armigera (Hübner), and the western bean cutworm Striacosta albicosta (Smith). These species might interact and compete by food, once they share the same feeding guild, the maize ear. The cannibalism/predation behavior is also present during the larval stage for most of these species, with exception of S. albicosta, which enhance the intraguild competition of them. In addition to this changing agroecosystem, which offer a range of host plant to these species, the recent identification of H. armigera in American continent, and also the dispersion of S. albicosta in the USA, have been raised several concerns involving the impact of larval interactions in the integrated pest management (IPM) and insect resistance management (IRM). Also, the complex management of these species, along with the adoption of transgenic (Bt) maize hybrids, the sub doses of Bt, and the resistance management strategies, involving refuge strategy, take the importance of noctuids behavior to other level. Understanding the larval behavior and interaction, firstly on non-Bt maize, is essential to find the possible prevalence of species in agricultural crops, and to obtain successes in IPM and IRM. In this sense, this study aimed to evaluate the intraguild interaction among larvae of different species and their movements, throught behavioral studies on non-Bt maize. Initially, the intraguild interaction of S. frugiperda, H. zea and S. albicosta was investigated under laboratory and field conditions in the USA, using several interaction scenarios with arenas and maize tissue. In another study in laboratory, it was assessed the attack and defense movements between S. frugiperda and H. zea in different interaction scenarios, with and without food, aiming to build an ethogram. A study between the allopatric species, H. zea and H. armigera, was carried out involving their intraguild interactions on maize crops, in intraspecific and interspecific scenarios, as well the larval occurrence and proportion in three regions of Brazil. Finnally, the intraguild interaction of S. frugiperda with the Helicoverpa spp. was investigated on scenarios in laboratory and field conditions, on maize in Brazil. A video-tracking system was utilized to characterize larval behavior parameters resulted from interactions. Regarding the first study, S. frugiperda presented advantage in interaction with the other species, while S. albicosta has disadvantage in the interactions. In the second study, S. frugiperda exhibited a predominance of defensive movements when competing against H. zea in the same instars. On the opposite, larvae of H. zea showed distinguished aggressive movements, with cannibalism and predation occurring in interactions between 6th instar with opponents in 4th instar. In the following study, H. zea was predominant in Rio Grande do Sul and central of Brazil. In western Bahia, the species was predominant in the first crop season, being the relation H. zea: H. armigera more similar in the second crop season. Larvae of H. zea presented advantage in interaction with H. armigera. The agroecosystem plays an important role in the species occurrence and impacts the interactions and prevalence of them in the environment. Larvae of S. frugiperda presented competitive advantage on Helicoverpa spp. Frequency and feeding time period in food of S. frugiperda larvae were negatively affected by interactions. Larvae of S. frugiperda moved shorter distances compared to H. zea.

<div>Forensic entomology is a subdiscipline of entomology that involves the use of insect behavior and developmental data to aid in criminal investigations. Genetic data has become increasingly important to the field as there has been a push for DNA-based species identification methods of forensically relevant insects. Genetic data can also elucidate population structure and relatedness of these insects, and such knowledge can contribute to the development of more specific datasets for insects in different regions. The first study presented here investigated the phylogenetics of sister species <i>Lucilia cuprina</i> and <i>Lucilia sericata</i> to identify possible subspecies divisions and issues with DNA-based identifications in the United States. The initial aim of this study was to identify genetic differences between specimens of <i>L. cuprina</i> that preferred live versus carrion flesh. Flies collected from Indiana, USA and South Africa were sequenced and analyzed. Upon sequencing of the genes <i>COI, Period,</i> and <i>28s,</i> our results indicated that <i>L. cuprina</i> from Indiana possess a unique combination of nuclear and mitochondrial haplotypes that suggest a unique lineage, possibly indicating modern hybridization with <i>L. sericata. </i>The inability of both nuclear and mitochondrial genes to distinguish between <i>L. cuprina</i> and <i>L. sericata</i> raises questions about the capabilities of DNA-based species identifications within this genus. Additionally, the inability of these genes to distinguish between specimens that preferred live versus carrion flesh highlights a need for continued research of these behavioral differences. The second study presented here investigated the population structure and relatedness of house flies in the American southwest in relation to a civil lawsuit where neighbors of a poultry farm alleged that flies were emanating from the farm to their homes. <i>Musca domestica</i> (house fly) specimens were collected from the chicken farm and from locations in varying directions and distances from the farm. Amplified fragment length polymorphism (AFLP) analysis was performed and the data were used in a number of analyses. Population reallocation simulations generally indicated that samples from different locations were not genetically different enough from other locations to allocate to their true origin population over others. Kinship analysis showed differences in samples collected in a later season that indicate a genetic bottleneck over time. Population structure analysis indicated the presence of two intermixing genetic populations in the dataset. AMOVA revealed that the majority of genetic variation laid within, rather than among, populations. A Mantel test revealed no significant correlation between genetic and geographic distances. These results indicate that the <i>M. domestica</i> population in this region of southwestern America is large and intermixing, with no clear genetic distinctions between specimens collected at the poultry farm versus the surrounding locations. In regard to the civil lawsuit, it was not possible to conclude that the flies did not emanate from the poultry farm. In a broader perspective, these data can be utilized to develop pest management strategies in this region. Overall, the data from both studies presented here will be useful to forensic investigations, development of more specific and detailed data and identification techniques, and pest control measures.</div>

Acoustic communication in orthopterans and anurans provides a suitable model system for studying the evolutionary mechanisms of sexual selection mainly because males use acoustic signals to attract females over long distances for pair formation. Females use these signals not only to localize conspecific males but also to discriminate between potential mates. Investigations on the effect of sexual selection on acoustic signals requires an understanding of how female preferences for different features of the acoustic signal affect male mating success under ecological constraints in wild populations. The effect of female preferences on male mating success depends on the mate sampling strategy that females employ to search for potential mates. Despite its relevance, female mate sampling strategies based on male acoustic signals have rarely investigated in orthopterans and anurans, especially in the field. Considering the elaborate knowledge of the role of sensory physiology in female phonotaxis behaviour and characterization of the male acoustic signal, I used the field cricket species Plebeiogryllus guttiventris as a model system in this study. In this thesis, I first investigated the ecology of callers in wild populations. I then investigated female mate sampling strategies by incorporating relevant information on the ecology of signalers and the sensory physiology of receivers. Amount of calling activity is a strong determinant of male mating success in acoustically communicating species such as orthopterans and anurans. While many studies in crickets have investigated the determinants of calling effort, patterns of variability in male calling effort in natural choruses remain largely unexplored. I therefore investigated the spatio-temporal dynamics of acoustic chorusing behaviour in a wild population. I first studied the consistency of calling activity by quantifying variation in male calling effort across multiple nights of calling using repeatability analysis. Callers were inconsistent in their calling effort across nights and did not optimize nightly calling effort to increase their total number of nights spent calling. Next, I investigated calling site fidelity of males across multiple nights by quantifying movement of callers. Callers frequently changed their calling site across calling nights with substantial displacement but without any significant directionality. Finally, I investigated trade-offs between within-night calling effort and energetically expensive calling song features such as call intensity and chirp rate. Calling effort was not correlated with any of the calling song features, suggesting that energetically expensive song features do not constrain male calling effort. The two key features of signaling behaviour, calling effort and call intensity, which determine the duration and spatial coverage of the sexual signal, are uncorrelated and function independently Acoustic signal variation and female preference for different signal components constitute the prerequisite framework to study the mechanisms of sexual selection that shape acoustic communication. Despite several studies of acoustic communication in crickets, information on both male calling song variation in the field and female preference in the same system is lacking for most species. First, I quantified variation in the spectral, temporal and amplitudinal characteristics of the male calling song in a wild population, at two temporal scales, within and across nights, using repeatability analysis. Carrier frequency (CF) was the most repeatable call trait across nights, whereas chirp period (CP) had low repeatability. I further investigated female preferences based on song features with high and low repeatability (CF and CP respectively). Females showed no consistent preferences for CF but were more attracted towards calls with higher rates (shorter CP). I also examined the effect of signal intensity, which is known to play a critical role in female phonotaxis behaviour, on female preferences for faster calls. Females preferred louder calls over faster ones, implying a dominant role for signal intensity in female evaluation of potential mates based on acoustic signals. Call intensity was also the only signal feature that was positively correlated with male size. In the final chapter, I investigated female mate sampling strategies based on acoustic signals using both theoretical and empirical approaches. Analytical models of mate sampling have demonstrated significant differences in individual fitness returns for different sampling strategies. However these models have rarely incorporated relevant information on the ecology of signalers and the sensory physiology of receivers. I used simulation models to compare the costs and benefits of different mate sampling strategies by incorporating information on relative spacing of callers in natural choruses and the effect of signal intensity on female phonotaxis behaviour. The strategy of mating with males that were louder at the female position emerged as the optimal sampling rule in the simulations. When tested empirically in the field using callers in natural choruses, females seemed to follow the optimal strategy of mating with males that were perceived as louder at their position.

Social insects are the most dominant terrestrial fauna for the last 50 million years. This tremendous ecological success is accompanied by the fact that sociality has evolved multiple times independently and achieved highest degree of complexity in insect lineages. The remarkable social organization found in insect societies is the result of finely balanced cooperation and conflict among the colony members. A typical hymenopteran colony is characterised by one or a few queens monopolizing reproduction and several sterile workers co-operatively raising brood and performing colony activities. The colonies are often conceptualized as superorganisms where groups of cooperative workers are compared with organs in the body, each of which accomplish a particular task like brood care, foraging and defence. The choice of tasks is often regulated by a systematic age polyethism. As the queens monopolize reproduction, they serve as the sole suppliers of eggs in the colony. Therefore, loss or death of the queen creates a crucial void which exposes the colony to potential reproductive conflict for the position of egg-layer. This crisis is expected to be extreme in monogynous colonies. The situation is rescued only after a new queen is established, and the whole process is known as queen succession. I am interested in this crisis management, and my thesis deals with potential and realized conflicts associated with queen succession and behavioural strategies involved in resolution of these conflicts. The queen can be replaced in two ways - either by a newly eclosed specialized reproductive individual, which happens in highly eusocial hymenopterans, or by an existing member of the colony (worker), as it happens in primitively eusocial hymenopterans. Unlike in highly eusocial species, the workers of primitively eusocial species retain their ancestral capability of mating and activating ovaries to produce both sons and daughters, which makes them suitable for taking up the role of queen in their lifetime. Hence, primitively eusocial species provide a unique situation where loss or death of the queen might result in severe reproductive conflict as the queen can be replaced by any one of the existing workers. Strictly monogynous colonies of the tropical primitively eusocial wasp Ropalidia marginata provide ideal opportunities to study the reproductive conflict and its resolution associated with queen succession because the queen is frequently replaced by one of her nestmates resulting in a serial polygyny. These queens have highly variable tenures of queenship ranging from seven to over 200 days, which, together with the fact that they are replaced by a variety of relatives such as daughters, niece and cousins, suggests a potential reproductive conflict with variable degrees of complexity. I have divided my thesis in three parts which are as follows -Natural queen turnover: Previous works from this lab have tried to characterize the queen succession in R. marginata colonies by experimentally removing the queen from the colony. As this design involves the experimenter intervening at a random point of the colony cycle, the colony might not respond in the similar way as it might have done to a natural succession necessitated by loss or death of the queen. But rarity and unpredictability of natural queen turnovers made them difficult to study. Therefore, in this section, we gathered a dataset of long-term and opportunistic quantitative behavioural observations on eleven natural queen turnovers and compared them with available data on queen removal experiments. All our queen removal experiments resulted in a hyper-aggressive potential queen who gradually reduced her aggression, activated her ovaries and went on to become the unanimously accepted new queen of the colony if the original queen was not returned. Here we found a similar phenomenon in natural colonies where a single un-challenged potential queen took over the colony as new queen after the old queen was lost, died or was driven out of the colony. In some of the natural colonies, the transition was preceded by aggression shown by the potential queens towards their nestmates including the queens, which indicates that they might have pre-empted the transition. The potential queens in natural colonies started laying eggs much faster than in experimental colonies suggesting their physiological readiness for the transition. How does a colony respond to a declining queen?: As we could show that some of the potential queens might perceive the upcoming queen turnover, a fair prediction would be that they sense it through the declining fertility status of the queens. Therefore, we tried to ex-perimentally induce situations where the queen appears to be declining, expecting that it might lead to a queen turnover. The growing evidence suggests that R. marginata queen maintains her status by applying a pheromone on the nest surface by rubbing the tip of her abdomen. We knocked down the nest to deny the queen the surface for applying her pheromone, and argued that the queen would be overthrown as the workers would sense her as infertile. To our surprise, the queen maintained her status in six out of seven colonies by applying her pheromone on the entire surface of the cage. However, the effectively insufficient concentration of pheromone elicited aggression from workers towards the queen, and the queen retaliated back with aggression. These results suggest that the pheromone, being an honest signal of fertility, is extremely important for the queen for maintaining her reproductive monopoly, and the workers are able to perceive the decline of the queen from her pheromone. Queen-successor conflict over access to reproduction: Here we more explicitly looked at the potential reproductive conflict between the queen and her successor over access to direct reproduction. We used the theory of parent-offspring conflict proposed by Robert Trivers (1974) as the conceptual framework and adapted it to unravel the pat-tern of queen-successor conflict in R. marginata colonies. According to this idea, we expected that there should be a pre-conflict zone where the queen and the successor both would agree that the queen should continue to reproduce, followed by a conflict zone where the successor would try to takeover but the queen would hang on, finally followed by post-conflict zone where they both would agree that the successor should reproduce. To test this expectation, we maintained the queen and the potential queen on either side of a wire-mesh partition, each with randomly chosen half of the workers. It allowed the potential queen (successor) to establish herself and then we reintroduced the queen to her side of the mesh daily till the queen gave up. We could behaviourally characterise all three zones which always appeared in the expected sequence. The pre and post-conflict zones had no aggressive interaction between the queen and the potential queen, whereas the conflict zone was characterized by aggressive falling fight between them. This is our first success in experimentally creating overt conflict between the queen and her successor. Overall we can say, that the queens and the potential queens of R. marginata show great behvioural plasticity which might have been shaped by natural selection as an adaptation for conflict resolution. We could show that the potential queens sometimes can predict the upcoming transition and pre-pare themselves accordingly, whereas they can also react to an experimentally created sudden loss of queen by hugely elevating their aggression. The docile queens, on the other hand, maintain their reproductive monopoly by a pheromone, which is essentially a feature of highly eusocial species. But these docile queens have not lost their capability to show aggression and can use that to complement the insufficient concentration of her pheromone. This and the behaviour of potential queens in their establishment phase are strongly reminiscent of typical primitively eusocial species. We conclude that Ropalidia marginata is, perhaps, a particularly advanced primitively eusocial hymenopteran situated on an evolutionary continuum from primitive to highly eusocial species.