Estudio de la muerte celular por Apoptosis en el ovario de Columba maculosa y Columbina picui (Aves:Columbidae) durante el Ciclo Reproductivo Anual. Study of cell death by apoptosis in the ovary of Columba maculosa and Columbina picui (Aves: Columbidae) during the reproductive annual cycle.

En este trabajo se analizarán las características estructurales, cuantitativas y el proceso de muerte celular por apoptosis en el ovario de C. maculosa y C.picui con el objetivo de aportar conocimientos básicos a la biología reproductiva de estas aves. Cincuenta hembras adultas de cada especie se capturarán en el Dpto. Río Primero (Pcia. de Cba), R. Argentina, durante el ciclo reproductivo 2011-2012. Las muestras de ovarios se fijaran en Formalina Neutra pH 7.0, procesarán con la técnica de inclusión en parafina y colorearan con Hematoxilina /Eosina y Reacción Nuclear de Feulgen. Cinco muestras serán utilizadas para la determinación de muerte celular por apoptosis con la técnica de TUNEL. Se estudiarán las características morfohistológicas del ovario de C.maculosa y C. picui e identificarán, categorizarán y cuantificarán los folículos atrésicos no bursting (folículos atrésicos previtelogénicos y vcitelogénicos pequeños que conservan la integridad de la pared folicular) y bursting (folículos vitelogénicos mayores de 2 mm que liberar el contenido folicular por ruptura de la pared folicular) durante el ciclo reproductivo anual. Mediante la marcación de ADN fragmentado, se revelará la muerte celular por apoptosis en las células granulosas de los folículos atrésicos. Se compararán las semejanzas y diferencias estructurales y cuantitativas y el proceso de muerte celular en los folículos regresivos entre las dos especies. Los resultados de este trabajo representarán un importante aporte al conocimiento de la atresia folicular como así también a la muerte celular, un proceso estrechamente asociado a la misma, aún poco estudiado en el ovario de las aves. In this work we analyse the structural, quantitative and the process of the cell death by apoptosis in the ovary of Columba maculosa and Columbina picui in order of providing basic knowledge of reproductive biology of these birds. Fifty adult female of each species will be caught in the Río Primero (Pcia. Cba) R.Argentina, during 2011 - 2012. The ovarian samples will be fixed en neutral buffered Formalin (pH 7.0), processed with the technique of inclusion in paraffin and stained with Haematoxylin - Eosin, and nuclear Reaction of Feulgen. Five samples will be used to reveal cell death by apoptosis with the technique of TUNEL. Will be studied the morphohistological characteristics of ovary of both species, and identify, categorize and quantify the atretic follicles over the cycle, the non-bursting (pre-vitellogenic follicles and vitellogenic small < 2 mm, involution an without follicular rupture) and bursting (vitellogenic follicle > 2 mm in the which the yolk falls into the peritoneum due to rupture of with out put of the wall follicular). Will be revealed DNA fragmentation in the granulosa cells of the atretic follicles. Will compared the similaritues and differences in structure and quantitative and the cell death process by apoptosis in the regresive follicles, between the two species. The results of this study represent an important contribution to the knowledge of follicular atresia as well cell death a process closely associated with it and still poorly studied in the ovary of birds.

Formação das castas no gênero Melípona (Illiger, 1806)

The present work is destinated to prove that the castes : workers and queens, in Melipona bees are due to genetic factors and not to differences in food. 2) Material used: Hives of Melipona quadri-fasciata anthidioides (Lep. 1836), M. schenki schenki (Gribodo, 1893), M. fasciata rufiventris (Lep. 1836), M. quadri-fasciata vicina (Lep. 1836), M. marginata marginata (Lep. 1836), Apis mellifera (L. 1758). 3) It should be pointed out that in Melipona bees there are no royal cells for the queens, but all the cells are of the same size independently of being destinated for workers, queens or drones. The numerous queens which are born are killed soon after emerging from their cells. 4) Changes of feeding in quality and in quantity caused no variation of castes. The only variable factor is the size, which becomes bigger when the bee is well nourished. 5) The offsprings of 5 hives were examined : 3 of M. quadri-fasciata anthidioides (n.o 1, n.o 2 and n.o 3), 1 of M. quadri-fasciata vicina (n.o 4) and 1 of M. marginata marginata (n.o 5). Combs of about 40 cells were taken into laboratory and the type of bee registered immediately after emerging. The results of the counts were: BOX COMB WORKER QUEEN PERCENTAGE Σ X2 to 12,5% Nº 1 1th 69 8 10,4% 0, 3139 " 1 2nd 144 18 11,1% 0, 2856 " 2 1th 52 8 13,3% 0, 0384 " 3 1th 45 10 18,2% 1, 6736 " 4 1th 56 4 6,7% 1, 8686 " 4 2nd 29 4 12,1% 0,00432 Σ X2 to 25% " 5 1th 34 14 29,2% 0,44444 "5 2nd 83 27 24,5% 0, 0121 In the 4 first boxes there is a percentage of 11,63% queens and in the last there is a percentage of 25,95%. 6) These percentages are very near two genetical ratios: 12,5% or 7:1, and 25% or 3:1, which correspond to a trifactorial and a bifactorial back-cross. Carrying out a X² test no significant deviations were found ( X² to 12,5% and to 25% and table 1 to 4). 7) We suppose that the formula for the queen in the first case (11,65%) is: AaBbCc. Since the Melipona bees are arrhenotokous hymenopteres, the drones are haploid and may have any one of the following eight formulas, corresponding to the gonic segregation of the queem : ABC, ABc, Abc, Abc, AbC, aBC, aBc, abC, abc. Anyone combination of these males with the queen will give a segregation of 7 workers to 1 queen, since there is always only one triple heterozygote among the eight possible segregates (table 5). 8) In order to explain the second case, it is suffient to assume that in this species there are only two pairs of factors, the queen being the double heterozygote : AaBb, while the drones may have any one of the following constitutions: AB, Ab, aB and ab. Workers are again all diploids which are homozygous for one or both factors, for instance: AABB, AABb, AaBB, aaBb, AAbb, etc. (table 6). 9) It is suggested that the genus Melipona is an intermediary type between the solitary bees, where all females are fertile independently of their feeding, and the genera Apis and Trigona, where without special feeding all females are born sterile, while only specially fed females develop into fertile queens. 10) No speculations are put forward with regards to the evolutionary mechanism which may have been responsible for the development of the genetical determination of castes in Melipona, since it seems advisable point to extend the studies to other insects with complicated caste systems.

Uma espécie nova de Schwarzula da Amazônia (Hymenoptera, Apidae, Meliponini)

Schwarzula coccidophila sp. nov., a tiny Amazonian stingless bee, that attends scale insects (Cryptostigma Ferris, 1922, Coccidae) in its nest, is described. It is distinguished from Schwarzula timida (Silvestri, 1902), the only other species of the genus, mainly by the malar area longer than diameter of 3rd flagellomere, and the denser plumose pilosity. Additional records of S. timida is presented.

Variation morphogeometrics of Africanized honey bees (Apis mellifera) in Brazil

The morphometrics of the honey bee Apis mellifera L., 1758 has been widely studied mainly because this species has great ecological importance, high adaptation capacity, wide distribution and capacity to effectively adapt to different regions. The current study aimed to investigate the morphometric variations of wings and pollen baskets of honey bees Apis mellifera scutellata Lepeletier, 1836 from the five regions in Brazil. We used geometric morphometrics to identify the existence of patterns of variations of shape and size in Africanized honey bees in Brazil 16 years after the classic study with this species, allowing a temporal and spatial comparative analysis using new technological resources to assess morphometrical data. Samples were collected in 14 locations in Brazil, covering the five geographical regions of the country. The shape analysis and multivariate analyses of the wing allowed to observe that there is a geographical pattern among the population of Apis mellifera in Brazil. The geographical variations may be attributed to the large territorial extension of the country in addition to the differences between the bioregions.

Uma estação biológica para o estudo dos mosquitos e dos outros animaes silvestres relacionados com a febre amarela

The outbreak of the jungle or forest yellow fever, through the adapta¬tion, quite recently of the yellow fever virus o the forest mosquitoes, brou¬ght the necessity of ecological researches on hese mosquitoes, as well as on the wild animals they bite, some of them being susceptible to the desease. This has been done by the special yellow fever Service of the State of Sao Paulo, in a special Biological Station in Perús, São Paulo, which has been built in the midst of the jungle. This station was made with plain materials, and covered with straw, but was confortable enough for the technical work, i nthe early months of 1938. During the months in which the investigations were being carried on, the following interesting results were obtained: 1. As we have already pointed out in other places, the forest mosquitoes biting us during daytime, are always new born insects, having not yet sucked blood, as it is the general rule with all mosquitoes, and therefore also, with the anopheles and stegomyia, and this explains why nobody gets malaria or yellow fever, transmitted by anofeles or by aedes aegypti during the day. We think therefore, the jungle yellow fever, got during daytime is not due to the infected jungle or forest mosquito biting, but to infection through the human skin coming into close contact with tre virus, which the forest mosquitoes lay with their dejections, on the leaves of the trees where they remain sitting du¬ring the day. 2. As it is the rule with anopheles, stegomyia and other mosquitoes, the insects once having sucked blood, take nocturnal habits and, therefore, bite us, only during the night, so it happens with the forest mosquito, and insects with developped eggs and blood in stomach have been caught within the sta¬tion house, during the night. During the day, these mosquitoes do not bite, but remain quite still on the leaves of the trees, in the damp parts of the woods. 3. Jungle or forest mosquitoes can easely bite wild animals, some with more avidity then ethers, as it has bee npointed out to the opossum (didei-phis) and other animals. They also bite birds having very thin skin and only exceptionally, cold bloods animals. 5. Is has hot been possible to ascertain how forest mosquitoes are able to live, from onde season to another, through winter, when temperature drops near and even below zero. They have not been found in holes of the terrain, of trees and of animals, as it is the rule in cold countries. During winter, in the forest, it is possible to find larvs in the holes of bambus and trees full of water. As wild animals do not harbour the yellow fever virus for a long time in their body, it is diffcult to explain how the desease lasts from one season to another. Many ecological features on the mosquito, remains yet to be explained and therefore it in necessary to go on with the investigations, in bio¬logical stations, such as that one built up in Perús, São Paulo.

Atividades da estação biológica de Perús

The author who was appointed entomologist of the Biological Station in Perus, São Paulo, describes in this paper, the kind of work he has been doing there. He begins with a description of the organization of the Station and of the routine work as it was daily carried on there, by himself and his staff, during nearly 6 months. During the day as well as during the night, captures of jungle were made in the forest and the same was done by night, in the Station House chiefly when the athmosphere was damp, just before, during, or after a rain. There was also an intensive search for foci of mosquitoes' larves in the bromelias, in holes, in trees and in the soil. The larves found in these breeding places were brought to a larvarium established in the forest in a place close to the station where they were bred in holes of bambus which were very suitable for them. During daytime, only new hatched mosquitoes have been captured, but during the night it has been possible to catch, inside the Station house, many female mosquitoes, with developped eggs, so confirming Aragão's opinion, that mosquitoes biting during the day are always, newly hatched ones. Some species of Sabetini were captured only inside the Biological Station House, during the night. The habits of the following species were subjected to more accurate investigations. Aedes scapularis, Aedes leucocelaenus, Lutzia braziliae, Culex (Carolia) iridescens, Orthopodomyia albicosta, Goeldia palidiventer, Joblotia compressum, Wyeomyia longirostris, Sabetoides intermedius, Limatus durhami. The conditions of the temperature of the Station, did not permit the authour to obtain breedings of Aedes aegypti in the larvarium of the Station, even during he summer months. A great diminuitions of species of the jungle mosquitoes was observed, from January till June, that is, when temperature gets lower and lower. The author has made the interesting observation that some species of mosquitoes (Joblotia and Limatus), must take a meal of flowers or bee honey before they suck blood. A list of the mosquitoes captured during the months of February to June, in the Station is given.

On the applicability of mathematics : philosophical and historical perspectives

The present thesis is a contribution to the debate on the applicability of mathematics; it examines the interplay between mathematics and the world, using historical case studies. The first part of the thesis consists of four small case studies. In chapter 1, I criticize "ante rem structuralism", proposed by Stewart Shapiro, by showing that his so-called "finite cardinal structures" are in conflict with mathematical practice. In chapter 2, I discuss Leonhard Euler's solution to the Königsberg bridges problem. I propose interpreting Euler's solution both as an explanation within mathematics and as a scientific explanation. I put the insights from the historical case to work against recent philosophical accounts of the Königsberg case. In chapter 3, I analyze the predator-prey model, proposed by Lotka and Volterra. I extract some interesting philosophical lessons from Volterra's original account of the model, such as: Volterra's remarks on mathematical methodology; the relation between mathematics and idealization in the construction of the model; some relevant details in the derivation of the Third Law, and; notions of intervention that are motivated by one of Volterra's main mathematical tools, phase spaces. In chapter 4, I discuss scientific and mathematical attempts to explain the structure of the bee's honeycomb. In the first part, I discuss a candidate explanation, based on the mathematical Honeycomb Conjecture, presented in Lyon and Colyvan (2008). I argue that this explanation is not scientifically adequate. In the second part, I discuss other mathematical, physical and biological studies that could contribute to an explanation of the bee's honeycomb. The upshot is that most of the relevant mathematics is not yet sufficiently understood, and there is also an ongoing debate as to the biological details of the construction of the bee's honeycomb. The second part of the thesis is a bigger case study from physics: the genesis of GR. Chapter 5 is a short introduction to the history, physics and mathematics that is relevant to the genesis of general relativity (GR). Chapter 6 discusses the historical question as to what Marcel Grossmann contributed to the genesis of GR. I will examine the so-called "Entwurf" paper, an important joint publication by Einstein and Grossmann, containing the first tensorial formulation of GR. By comparing Grossmann's part with the mathematical theories he used, we can gain a better understanding of what is involved in the first steps of assimilating a mathematical theory to a physical question. In chapter 7, I introduce, and discuss, a recent account of the applicability of mathematics to the world, the Inferential Conception (IC), proposed by Bueno and Colyvan (2011). I give a short exposition of the IC, offer some critical remarks on the account, discuss potential philosophical objections, and I propose some extensions of the IC. In chapter 8, I put the Inferential Conception (IC) to work in the historical case study: the genesis of GR. I analyze three historical episodes, using the conceptual apparatus provided by the IC. In episode one, I investigate how the starting point of the application process, the "assumed structure", is chosen. Then I analyze two small application cycles that led to revisions of the initial assumed structure. In episode two, I examine how the application of "new" mathematics - the application of the Absolute Differential Calculus (ADC) to gravitational theory - meshes with the IC. In episode three, I take a closer look at two of Einstein's failed attempts to find a suitable differential operator for the field equations, and apply the conceptual tools provided by the IC so as to better understand why he erroneously rejected both the Ricci tensor and the November tensor in the Zurich Notebook.

Capacitación en meliponicultura de la población de Poço Redondo (Sergipe, Brasil)

Con la deforestación, los incendios y la utilización indebida de los agrotóxicos, las abejas nativas de Brasil corren un serio peligro de extinción. Ello se debe a que estas abejas construyen sus nidos en los huecos de los troncos de los árboles y también a que sus fuentes de alimentación son cada vez más escasas. Por ello cada vez son más los que se dedican a la meliponicultura, de esta manera se conservan las abejas y además, se obtiene una renta extra. Mediante el presente proyecto se capacitó en meliponicultura a una muestra de la población de Poço Redondo, para que de esta manera se empiece a recuperar su entorno y también su situación económica actual. Para poder conseguirlo se llevó a cabo el diseño del curso mediante la documentación previa, el trabajo de campo y el análisis de los datos. El resultado fue la capacitación, con la que se formó y concienció ambientalmente a personas para que se dedicaran a la meliponicultura de manera profesional.

Presença de material eletrodenso sob a membrana vitelina do ovo de Scaptotrigona postica latr. (Hymenoptera, Meliponinae) durante a fase inicial da embriogenese

The present paper reports the presence of an electrondense material of unknown nature distributed under the viteline membrane of Scaptotrigona postica egg. The dorsal side layer is thicker then the ventral one. In eggs newly oviposited the material is gradually distributed inside the plasmalema invaginations. Later on (12 hs after oviposition) when a blastoderm is already formed around the egg, the material moves to the intercellular space. On the sequence of the development there was no more indication of the material under the membrane.

Ecological-economic modelling of interactions between wild and commercial bees and pesticide use

The decline in extent of wild pollinators in recent years has been partly associated with changing farm practices and in particular with increase of pesticide use. In this paper we combine ecological modelling with economic analysis of a single farm output under the assumption that both pollination and pest control are essential inputs. We show that the drive to increase farm output can lead to a local decline in the wild bee population. Commercial bees are often considered an alternative to wild pollinators, but we show that their introduction can lead to further decline and finally local extinction of wild bees. The transitions between different outcomes are characterised by threshold behaviour and are potentially difficult to predict and detect in advance. Small changes in economic (input prices) and ecological (wild bees carrying capacity and effect of pesticides on bees) can move the economic-ecological system beyond the extinction threshold. We also show that increasing the pesticide price or decreasing the commercial bee price might lead to reestablishment of wild bees following their local extinction. Thus, we demonstrate the importance of combining ecological modelling with economics to study the provision of ecosystem services and to inform sustainable management of ecosystem service providers.

Sociogenomics of Cooperation and Conflict during Colony Founding in the Fire Ant Solenopsis invicta.

One of the fundamental questions in biology is how cooperative and altruistic behaviors evolved. The majority of studies seeking to identify the genes regulating these behaviors have been performed in systems where behavioral and physiological differences are relatively fixed, such as in the honey bee. During colony founding in the monogyne (one queen per colony) social form of the fire ant Solenopsis invicta, newly-mated queens may start new colonies either individually (haplometrosis) or in groups (pleometrosis). However, only one queen (the "winner") in pleometrotic associations survives and takes the lead of the young colony while the others (the "losers") are executed. Thus, colony founding in fire ants provides an excellent system in which to examine the genes underpinning cooperative behavior and how the social environment shapes the expression of these genes. We developed a new whole genome microarray platform for S. invicta to characterize the gene expression patterns associated with colony founding behavior. First, we compared haplometrotic queens, pleometrotic winners and pleometrotic losers. Second, we manipulated pleometrotic couples in order to switch or maintain the social ranks of the two cofoundresses. Haplometrotic and pleometrotic queens differed in the expression of genes involved in stress response, aging, immunity, reproduction and lipid biosynthesis. Smaller sets of genes were differentially expressed between winners and losers. In the second experiment, switching social rank had a much greater impact on gene expression patterns than the initial/final rank. Expression differences for several candidate genes involved in key biological processes were confirmed using qRT-PCR. Our findings indicate that, in S. invicta, social environment plays a major role in the determination of the patterns of gene expression, while the queen's physiological state is secondary. These results highlight the powerful influence of social environment on regulation of the genomic state, physiology and ultimately, social behavior of animals.

The eusociality continuum

Eusocial societies are traditionally characterized by a reproductive division of labor, an overlap of generations, and cooperative care of the breeders' young. Eusociality was once thought to occur only in termites, ants, and some bee and wasp species, but striking evolutionary convergences have recently become apparent between the societies of these insects and those of cooperatively breeding birds and mammals. These parallels have blurred distinctions between cooperative breeding and eusociality, leading to calls for either drastically restricting or expanding wage of these terms. We favor the latter approach. Cooperative breeding and eusociality are not discrete phenomena, but rather form a continuum of fundamentally similar social systems whose main differences lie in the distribution of lifetime reproductive success among group members. Therefore we propose to array vertebrate and invertebrate cooperative breeders along a common axis, representing a standardized measure of reproductive variance, and to drop such (loaded) terms as ''primitive'' and ''advanced'' eusociality. The terminology we propose unites all occurrences of alloparental helping of kin under a single theoretical umbrella (e.g., Hamilton's rule). Thus, cooperatively breeding vertebrates can be regarded as eusocial, just as eusocial inverbrates are cooperative breeders. We believe this integrated approach will foster potentially revealing cross-taxon comparisons, which are essential to understanding social evolution in birds, mammals, and in sects.

In vitro antimicrobial assessment of Cuban propolis extracts

Propolis is a resinous mixture of different plant exudates collected by honeybees. Currently, propolis is widely used as a food supplement and in folk medicine. We have evaluated 20 Cuban propolis extracts of different chemical types, brown (BCP), red and yellow (YCP), with respect to their in vitro antibacterial, antifungal and antiprotozoal properties. The extracts inhibited the growth of Staphylococcus aureus and Trichophyton rubrum at low µg/mL concentrations, whereas they were not active against Escherichia coli and Candida albicans. The major activity of the extracts was found against the protozoa Leishmania, Trypanosoma and Plasmodium, although cytotoxicity against MRC-5 cells was also observed. The BCP-3, YCP-39 and YCP-60 extracts showed the highest activity against P. falciparum, with 50% of microbial growth (IC50) values of 0.2 µg/mL. A positive correlation between the biological activity and the chemical composition was observed for YCP extracts. The most promising antimicrobial activity corresponds to YCP subtype B, which contains acetyl triterpenes as the main constituents. The present in vitro study highlights the potential of propolis against protozoa, but further research is needed to increase selectivity towards the parasite. The observed chemical composition-activity relationship of propolis can contribute to the identification of the active principles and standardisation of this bee product.

Estudi comparatiu de l'algorisme basat en una colònia artificial d'abelles

La finalitat d'aquest projecte és la realització d'un estudi comparatiu de l'algoritme basat en una colònia artificial d'abelles, Artificial Bee Colony (ABC), comparat amb un conjunt d'algoritmes fonamentats en el paradigma de la computació evolutiva. S'utilitzarà l'eficàcia a l'hora d'optimitzar diverses funcions com a mesura comparativa. Els algoritmes amb els quals es comparara l'algoritme ABC són: algoritmes genètics, evolució diferencial i optimització amb eixam de partícules.

Conflict resolution and evolution of social structures in insect societies

In colonies of social Hymenoptera (which include all ants, as well as some wasp and bee species), only queens reproduce whereas workers generally perform other tasks. The evolution of worker's reproductive altruism can be explained by kin selection, which states that workers can indirectly transmit copies of their genes by helping the reproduction of relatives. The relatedness between queens and workers may however be low, particularly when there are multiple queens per colony, which limits the transmission of copies of workers genes and increases potential conflicts between colony members. In this thesis, we investigated the link between social structure variations and conflicts, and explored the mechanisms involved in variation of colony queen number in ants. According to kin selection, workers should rear the brood they are most related to. In social Hymenoptera, males are haploid whereas females (workers and queens) are diploid. As a result, workers can be up to three times more related to females than males in some colonies, where they should consequently favour the production of females. In contrast, queens are equally related to daughters and sons in all types of colonies and therefore should favour a balanced sex ratio. In a meta-analysis across all studies of social Hymenoptera, we showed that colony sex ratio is generally largely influenced by workers. Hence, the evolution of social structures where queens and workers are equally related to males and females may contribute to decrease the conflict between the two castes over colony sex ratio. Another conflict between queens and workers can occur over male production. Many species contain workers that still have the ability to lay haploid eggs. In some social structures, workers are on average more related to sons of queens than to sons of other workers. As a result, workers should eliminate worker-laid eggs to favour queen-laid eggs. We showed that in the ant Formica selysi, workers eliminate more worker-laid than queen-laid eggs, independently of colony social structure. These results therefore suggest that worker policing can evolve independently from relatedness, potentially because of costs of worker reproduction at the colony-level. Colony queen number is a key parameter that influences relatedness between group members. Queen body size is generally linked to the success of independent colony foundation by single queens and may influence the number of queens in the new colony. In the ant F. selysi, single-queen colonies produce larger queens than multiple-queen colonies. We showed that this association results from genes or maternal effects transmitted to the eggs. However, we also found that queens produced in colonies of the two social forms did not differ in their general ability to found new colonies independently. Queen body size may also influence queen dispersal ability and constrain small queens to be re-adopted in their original nest after mating at proximity. We tested the acceptance of new queens in another ant species, Formica paralugubris, which has numerous queens per colony. Our results show that workers do not discriminate between nestmate and foreign queens, and more generally accept new queens at a limited rate. To conclude, this thesis shows that mechanisms influencing variation in colony queen number and the influence of these changes on conflict resolution are complex. Data gathered in this thesis therefore constitute a solid background for further research on the evolution and the maintenance of complex organisations in insect societies.