Orientador: Odair Correa Bueno / Banca: Denise S. Scheepmaker / Banca: Ana Eugenia de Carvalho Campos / Banca: José Chaud Netto / Banca: Osmar Malaspina / Resumo: Interações insetos/micro-organismos são amplas e ocorrem das mais diversas maneiras, com as bactérias simbiontes de insetos desempenhando vários papéis, na maioria desconhecidos, na biologia do hospedeiro. O que se considerava apenas um único organismo eucariótico é na verdade um agregado de vários diferentes organismos, o que levou a uma mudança na maneira de se estudar esses organismos, culminando em uma abordagem mais holística. Dentro desse vasto mundo, relativamente pouco conhecido dos micro-organismos em associação com insetos, estão as bactérias do gênero Wolbachia (Classe Alphaproteobacteria, Ordem Rickettsiales), amplamente distribuídas nos artrópodes e transmitidas maternalmente, causadoras de diversas alterações reprodutivas no hospedeiro, sendo que sua ocorrência em populações naturais pode ser de grande interesse no controle biológico. A distribuição dessas bactérias em formigas é pouco explorada e existe carência de informações sobre a interação com formigas do gênero Solenopsis, que inclui espécies nativas da América do Sul. Este gênero possui espécies distribuídas de forma cosmopolita e no Brasil estão amplamente disseminadas, associadas preferencialmente a áreas de atividade humana. Este trabalho teve como objetivo analisar a diversidade genética da Wolbachia de amostras de ninhos de populações nativas de espécies do gênero Solenopsis, através do sequenciamento de cinco genes que compõe o multilocus de Wolbachia, além do gene wsp, a fim de caracterizar as cepas e estabelecer inferências filogenéticas entre elas. Além disso, testar as hipóteses de coevolução entre as cepas e as formigas e de recombinação entre as cepas encontradas. Com o sequenciamento e análises dos cinco genes que compõem o multilocus de Wolbachia (gatB, coxA, hcpA, ftsZ e fbpA), totalizando 2079 pb, destacam-se os seguintes resultados: i. o registro de 15 novas cepas; ii. o registro de 11 alelos... / Abstract: Insects/microorganisms interactions are broad and occurs in many different ways with symbiont bacteria of insects playing many roles, most unknown, on the biology if its host. What was considered a single eukaryotic organism is actually an aggregate of many different organisms which lead to a change in the way we study organisms, leading to a more holistic approach. Within this vast but relatively unknown world of microorganisms in association with insects are the bacteria of the genus Wolbachia (Class Alphaproteobacteria, Order Rickettsiales) widely distributed in arthropods and maternally transmitted, causing several reproductive alterations in the host, and their occurrence in natural populations being of great interest in the biological control of insects. Its distribution in ants is poorly explored and little is known about the interaction with the Solenopsis genus which includes native species from South America. This genus includes species cosmopolitan distributed and in Brazil they are widely distributed being preferentially associated with areas of human activity. This study aimed to analyze the genetic diversity of samples of nests from native populations of Solenopsis species infected by Wolbachia by sequencing the five genes comprising the Wolbachia multilocus, and also the wsp gene in order to characterize the strains and establish phylogenetic inferences between them. Furthermore test the hypothesis of coevolution between ants and its Wolbachia strains and recombination between found strains. With the sequencing and analysis of five genes comprising the Wolbachia multilocus (gatB, coxA, hcpA, ftsZ e fbpA) totaling 2079 bp the following results are highlighted: i. the record of 15 previously unknown new strains, ii. the record of 11 previously unknown new alleles, iii. phylogenetic relationship between the strains found here presents a polyphyletic pattern, indicative of the complexity of the evolutionary history of strains ... / Doutor

Orientador: Odair Correa Bueno / Banca: Osmar Malaspina / Banca: Terue Sadatsune / Banca: Maria Santina de Castro Morini / Banca: Ana Eugênia de Carvalho Campos Farinha / Resumo: As formigas podem adquirir bactérias presentes no ambiente e permitir que elas se desenvolvam nos seus tratos digestórios, transmitindo-as para as formas mais jovens. Estudos filogenéticos, com formigas do gênero Camponotus, apontam uma co-evolução das espécies de bactérias e de formigas, indicando também uma transmissão maternal. As formigas andarilhas, ou “tramp species”, podem ocorrer em diversos ambientes, entre eles os hospitais, podendo atuar como carreadores de bactérias patogênicas. Não existem na literatura trabalhos que relatem bactérias patogênicas no trato digestório de formigas que ocorrem em ambientes urbanos. O objetivo do presente trabalho foi o de verificar a existência de bactérias patogênicas no trato digestório de formigas que ocorrem em ambientes hospitalares, determinar se essas espécies poderiam funcionar como reservatórios de bactérias e comparar com as bactérias isoladas de formigas de laboratório. Os resultados obtidos indicaram que as formigas não apresentam bactérias patogênicas em seus tratos digestórios, mesmo sendo coletadas em ambiente hospitalar, porém estas podem eventualmente ser isoladas, não fazendo parte da microbiota natural dessas espécies. Bactérias patogênicas foram isoladas nos ninhos mantidos em laboratórios e também nas larvas das espécies Monomorium pharaonis e Monomorium floricola. Devido às diferenças morfológicas entre o trato digestório das operárias adultas e larvas, pode ser explicada a distinção encontrada entre a microbiota isolada. Foram realizados o levantamento da distribuição das espécies de formigas no Hospital das Clínicas da Faculdade de Medicina da UNESP de Botucatu, SP, e no Centro de Saúde de São Carlos, bem como o isolamento das bactérias presentes no trato digestório dos exemplares coletados. Como não foram isoladas bactérias patogênicas... (Resumo completo, clicar acesso eletrônico abaixo) / Ants can collect bacteria from environment and led them develop and reproduce in their digestive tracts, transmitting them to young forms. Phylogenetic studies with ants of Camponotus genera, pointed the coevolution between ant and bacteria species, indicating also maternal transmission. Tramp species are ant species that live in close association with man and occur in several environments, like hospitals. There is no works in literature about pathogenic bacteria isolated from digestive tract of ants collected in urban environments. The objective of present work were to verify the presence of pathogenic bacteria in digestive tract of ants collected at hospitals, and determine if these species can act as bacterial reservatories, comparing them with laboratory ants. The results obtained indicated that ants donþt presented pathogenic bacteria in their digestive tracts, even those ants collected at hospitalar environment. But, these bacteria could be eventually isolated, donþt being part of the natural gut microflora of these species. Pathogenic bacteria were also isolated from colonies maintained in laboratory conditions and from M. pharaonis and M. floricola larvae. Based at the differences between digestive tracts of adults and larvae workers, we could suppose that gut microflora founded would be distinct. A survey was made about ant species presents and their distribution at HC from UNESP Botucatu, SP and a survey was made at another Hospital, at São Carlos, SP. Ants collected at both hospitals were utilized in microbiological experiments to verify the gut microflora. Utilizing molecular procedures of DNA extraction and PCR (Polymerase Chain Reaction), we could sequence DNA from bacteria that were not isolated by commom microbiological methodology. Were utilized sterile ants from T. melanocephalum species... (Complete abstract, click electronic address below) / Doutor

Orientador: Maurício Bacci Júnior / Banca: Luiz Carlos Forti / Banca: João Batista Fernandes / Resumo: As formigas cortadeiras Atta sexdens, conhecidas popularmente como saúvas, são consideradas pragas agrícolas, e são controladas principalmente por inseticidas tóxicos, altamente inespecíficos, prejudiciais ao homem e outros animais, além de poluir o solo e a água. Estas formigas extraem seus nutrientes através da degradação da matéria vegetal, realizada por polissacaridases (especialmente pectinases), que estão presentes em seU fluido fecal e são produzidas pelo fungo simbionte Leucoagaricus gongylophorus. Assim, a inibição de pectinase seria uma estratégia promissora para controlar as formigas cortadeiras na agricultura de modo mais específico. No presente trabalho, nós buscamos inibidores de pectinase do fluido fecal em várias partes de 22 espécies vegetais, ensaiando 126 extratos brutos, 99 frações ou subfrações e 41 substâncias puras isoladas destes vegetais. Nossos resultados indicam que (1) Cedrela fissilis (Meliaceae) contém inibidores da pectinase presente no intestino das formigas. (2) O flavonóide (-)-catequina foi identificado como composto majoritário ou puro de todas as frações ativas dos extratos de Cedrela fissilis. (3) O flavonóide (-)-catequina, purificado de Cedrela fissilis, assim como extratos desta planta contendo majoritariamente (-)-catequina, inibiram a atividade de pectinase contida no fluido fecal, mas não a atividade de pectinase contida nos jardins de fungo, que provavelmente contém inibidores da ação da (-)-catequina. Assim, sugerimos que o jardim de fungo funciona como uma barreira protetora para as formigas / Abstract: The leaf-cutting ants Atta sexdens are agricultural pests controled by chemicals which are toxic, not specific and harmful to man, other animal and water resources. They feed on the degradation products of plant material which they cut, harvested to their nest and offer to a symbiotic fungus Leucoagaricus gongylophorus, which grows on this plant material to form the so-called fungus garden. The fungus produces polysaccharidases (especially pectinase) which the ants concentrate in their intestine and secretes in the faecal fluid. Thus, pectinase inhibition would negatively affect the ants nutrition and could be a promising strategy for the control of leafcutters. In the present work, we have searched for pectinase inhibitors in 126 extracts, 99 fractions or sub-fractions and 41 compounds which have been isolated from 22 plant species. Our results showed that (1) Cedrela fissilis (Meliaceae) contains inhibitors of the pectinase which is contained in the ants' faecal fluid. (2) (-)-Catequin, a flavonoid, was the major or the unique compound in every C. fissilis preparation inhibiting the pectinase activity of faecal fluid. (3) These preparations, however, have not inhibited the pectinase activity of the fungus garden. We suggest that the fungus garden acts as a barrier which protects the fungal enzymes against harmful compounds which may be introduced in the ants'nest. the pectinase activity of the fungus garden. We suggest that the fungus garden acts as a barrier which protects the fungal enzymes against harmful compounds which may be introduced in the ants'nest / Mestre

Orientador: Odair Correa Bueno / Banca: Jacques Hubert Charles Delabie / Banca: Benedito Cortês Lopes / Banca: Maria Izabel Souza Camargo / Banca: Edilberto Giannotti / Resumo: A bioecologia e o comportamento de Cyphomyrmex lectus e de Cyphomyrmex transversus (Formicidae: Myrmicinae: Attini) foram comparados. As colônias de C. lectus são pequenas (73,75 l 46,61 indivíduos) e nidificam no solo, enquanto as de C. transversus são maiores (249,1 l 240,4 indivíduos) e nidificam principalmente sob cocos caídos. O fungo miceliforme de C. lectus e o fungo leveduriforme de C. transversus são cultivados com os mesmos tipos de substratos animais e vegetais. Diariamente, dois picos de forrageamento ocorrem para C. lectus e apenas um para C. transversus. Preparação e incorporação do substrato no fungo são diferentes entre as duas espécies e alguns comportamentos revelaram-se próprios a cada uma. Descobriram-se microcavidades na cutícula da pró-pleura das duas espécies servindo para alojar bactérias simbiontes. A morfologia larval é muito similar entre as duas espécies; no entanto, a morfometria revelou cinco instares larvais para C. transversus e somente quatro para C. lectus. A morfologia do aparelho reprodutor das fêmeas é muito similar entre as duas espécies. Os ovários com três ovaríolos cada um são do tipo politrófico, bem desenvolvidos e funcionais para a rainha, mas reduzidos para a gine e a operária. O desenvolvimento da espermateca é igual entre rainha e gine das duas espécies cujas colônias são monogínicas. A etologia comparada da rainha e da gine revelou que a primeira tem uma função social orientada para a reprodução, enquanto a gine tem um papel similar ao das operárias. O cuidado com os imaturos pelas operárias é semelhante nas duas espécies. Enfim, um excepcional caso de parasitismo múltiplo por vespas Acanthopria (Hymenoptera: Diapriidae) ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The ecology and behavior of Cyphomyrmex lectus and Cyphomyrmex transversus (Formicidae: Myrmicinae: Attini) were compared. The colonies of C. lectus are small (73,75 l 46,61 individuals) and nest in the ground, while those of C. transversus are larger (249,1 l 240,4 individuals) and nest mostly under dry coconuts on the floor. The myceliform fungus of C. lectus and the yest-like form fungus of C. transversus are cultivated with the same types of animal and vegetal substrates. Each day, there are two peaks of foraging for C. lectus and only one for C. transversus. Preparation and incorporation of the substrate in the fungus are different between the both species and some behaviors were shown to be characteristic to only one or the other species. Micro-holes that host symbiotic bacteria were found in the cuticle of the propleura of the two species. The larval morphology is very similar between them. The morphometric analysis showed yet five larval instars for C. transversus and only four for C. lectus. The morphology of the reproductive system of the females is very similar between them. The ovaries of three ovarioles belong to the polytrophic type, are well developed and functional in the queen, but reduced in the gyne and the worker. The spermatheca development is the same in the gyne and the queen when colonies are monogyne. The compared behavior of the queen and the gyne showed that the first has a social function oriented to the reproduction, while the gyne has a function similar to a worker. The workers take care of immatures similarly in the both species... (Complete abstract click electronic access below) / Doutor

Made available in DSpace on 2014-06-11T19:30:16Z (GMT). No. of bitstreams: 0 Previous issue date: 2004Bitstream added on 2014-06-13T20:00:17Z : No. of bitstreams: 1 campos_mcg_me_rcla.pdf: 269650 bytes, checksum: 2539c2d250c795c665b585ed1efd4558 (MD5) / Há muito tempo vem sendo constatada a ocorrência de formigas em ambientes urbanos; esses insetos podem acarretar danos em residências e em vários utensílios de uso domésticos, além de causar sérios problemas, pois se tornam um perigo à saúde pública quando a infestação se dá em hospitais. Métodos eficientes de controle de formigas urbanas envolvem um manejo adequado do ambiente e a utilização de iscas atrativas tóxicas. Estas funcionam como fonte de alimento e, desta forma, podem provocar intensa competição entre as várias espécies. As formigas possuem uma enorme variedade de estratégias ofensivas e defensivas na procura de alimento e proteção do ninho. A maior parte dessas agressões pode ser vista como uma série de comportamentos que atuam como técnicas competitivas. Competição compreende a demanda ativa por dois ou mais indivíduos da mesma espécie (competição intraespecífica) ou membros de duas ou mais espécies de mesmo nível trófico (competição interespecífica) por um recurso comum potencialmente limitado. Foram realizados seis testes em laboratório, com colônias de formigas urbanas, com o objetivo de estudar as interações comportamenatis na fonte de alimento. As colônias se encontravam em uma bandeja de plástico e foram providas de mel e larvas de besouro Tenebrio. O suprimento de água deu-se com um tubo de ensaio com algodão na extremidade livre. Os alimentos atrativos utilizados foram: uma mistura composta de fígado de boi desidratado, bolo, mel, e 1% de óleo de amendoim, e açúcar líquido diluído em água (75%). Para a realização dos testes foi utilizado um recipiente denominado de arena de forrageamento. Durante cada teste, os recipientes de criação das duas das espécies a serem estudadas foram conectados à esta arena, e as fontes de alimento foram colocadas no centro... / The attractive food used was a composed mixture of dehydrated ox liver, cake, honey, and 1% of peanut oil. The second food was a diluted liquid sugar in water (75%). For the accomplishment of these tests a container called arena of forage was used. During each test, the containers of creation of two of the species studied were connected to this arena, and the food source was placed in the center. After this procedure, the interaction behaviors of the ants were written down. Six tests of field were carried out where the attractive food was placed in fixed points in the studied environment close to the occurrence place of the ants species and it was taken notes of the behaviors of competition among all species. In accordance with the results of the laboratory tests, the order of domination among the four studied species was: Monomorium floricola, Monomorium pharaonis, Linepithema humile and Tapinoma melanocephalum. The two last ones were considered subordinated in relation to the competitive ability for interference, although they presented a bigger competitive ability for exploration. According to the final results of the tests carried through natural situations, it can be concluded that there is an effective competition for food among the different species of urban ants. However it is very difficult to carry on comments or observations about this process, since the sucession of different species of ants on food occurs in a short period of time and with a great rapidity.

The assemblage of bacteria and fungi associated with red imported fire ants (RIFA) Solenopsis invicta Buren was obtained from Hinds, Leake, and Madison Counties (location) along Natchez Trace Parkway in Mississippi. The sites were selected due to the limited presence of RIFA within the park and the more natural, undisturbed ecosystem. Active mounds containing soil, plant debris, and RIFA (substrate) were collected in March, July, and November of 2004 (time). Samples were processed according to standard microbiological protocols, and microorganisms identified using morphological, biochemical and molecular methods. A total of 71 bacteria (2324 isolates) and 50 fungi (1445 isolates) were obtained. The most common bacterium and fungus identified were Bacillus sp. B76(B)Ydz-zz, and Trichoderma aureoviride strain IMI 113135. The fungal entomopathogens Paecilomyces lilacinus and Metarhizium anisopliae var. anisopliae were isolated from mound soil, plant debris, and external tissues of the ants. Patterns of species richness, diversity, and evenness values across substrates were 71, 1.58, and 0.37 for bacteria, and 50, 1.11 and 0.28 for fungi, respectively. Total coefficient of community values for bacteria were 0.74 – 0.89 and 0.79 – 0.92 for fungi indicating uniform communities. No consistent trends were observed by comparing substrate, location, and sampling date. However, fungi species richness and diversity for ant external tissues were significantly higher than internal tissues of the ant. Selected bacteria and fungi were evaluated for their biological control and/or antagonistic potential in vitro and in situ. The most promising isolates studied in vitro included Paenibacillus sp. JA-08, Aspergillus terreus, and Aspergillus sp. HZ-35 with death rates on mound soil surface at 4.4, 5.0, and 4.8. The fungus Metarhizium anisopliae var. anisopliae strain LRC 211 had low death rate (1.8) on mound soil during in vitro trial but showed the greatest biocontrol potential during in situ evaluation. After 14 days in situ evaluation, the living RIFA extracted showed sluggish movement and the fungus was recovered from dead (48.3%) and living (33.3%) RIFA. Since the in situ trials were conducted only at one location and season, additional tests, including microscopic documentation of parasitism/pathogenicity, are needed to confirm the results of this study.

The ability to synchronize both the movements and activity rhythms of many interacting individuals is a major component of the repertoire of collective behaviour in social species. Among the many social animals on Earth, ants have some of the most spectacular forms of synchronized behaviour because of their eusociality. One example of this impressive behavioral synchronization in ants is the so-called short-term activity cycle, where the workers inside a nest will move together in rhythmic pulses of activity that are separated by moments of collective rest and inactivity. There are many aspects of short-term activity cycles that are poorly understood. The first chapter of this thesis makes the case for why ants are a useful taxonomic group for studying social synchronization and introduces the phenomenon of ant short-term activity cycles. In chapter 2, I study intraspecific variation in short-term activity cycles and show that colonies’ queens, number of brood items, and number of workers all influence aspects of colony-level activity oscillations. In chapter 3, I show that colonies are capable of synchronizing despite noise in the behaviour of individual ants, and that colonies can modulate between multiple collective rhythms. In the same chapter, I also devised a model to study the role of noise more generally in excitable systems that have properties similar to those in ant colonies. Chapter 4 examined the effect of 24-hour light cycles on circadian activity, and I found that colonies kept in conditions with constant light or darkness had weaker circadian rhythms than colonies in an alternating light/dark regime. However, colonies’ short-term activity cycles were not affected by the external light regime. In chapter 5, I provide evidence that having synchronized rhythms of rest and activity provides a functional benefit for colonies: workers can, on average, more fully inspect the inside of the nest when activity is synchronized because inactive ants aggregate into piles that active ants have difficulty penetrating. In chapter 6, I recount a history of the research into synchronized and rhythmic activity patterns inside ant nests and consolidate the new results from the previous chapters with the existing literature to create an up-to-date review of what we currently know about short-term activity cycles. The work compiled here thereby sheds new light on an enigmatic form of synchronized behaviour in ants, one of the most ecologically important groups of animals on the planet. / Thesis / Doctor of Philosophy (PhD)

The study was divided into three main sections. In the first section, temporal stability of genetic structure of red imported fire ant populations and effective population sizes were assessed with the help of microsatellites. The second part of the study was focused on the development of new microsatellite markers for the population genetics study of fire ants. The third part of the study was aimed at describing the association of phenotypic and genotypic characters of polygyne hybrid (Solenopsis invicta x richteri) fire ant in a colony and identifying the origin of polygyny in hybrid imported fire ant colony. Meridian, Yazoo City and Crystal Springs in Mississippi and Mobile, Alabama were selected for conducting the study. The overall FST estimates and the pairwise FST estimates between the populations of study, indicated little genetic differentiation and low spatial variation in the nuclear genetic structure of red imported fire ant. Gene flow estimates indicated that there is extensive movement between these populations. From 2006 to 2008 the FST estimates decreased and gene flow estimates increased, indicating that there is no equilibrium between genetic drift and migration in the populations studied. Effective population sizes ranged from 10 -17 in these populations.We have isolated and characterized an additional 11 polymorphic microsatellite loci in the invasive ant S. invicta from a population in Lauderdale County, Mississippi. The observed and effective number of alleles ranged from two to six (average 3.1818 ± S. E. 0.0486). Probability tests indicated significant deviations from the Hardy-Weinberg equilibrium at three loci. The polymorphism information content of the microsatellites ranged from 0.1482 to 0.6208.Identifying multiple inseminated queens in a colony and sequencing the critical region of Gp-9 locus (1600 base pairs) in individuals with the polygyne allele strongly indicate the association of phenotypic and genotypic characters of polygyny in a hybrid imported fire ant colony. A 100% match of Gp-9B and Gp-9b alleles and concordance in the critical amino acid substitutions of the Gp-9 locus in the hybrid fire ant with S. invicta Buren provides the evidence that polygyny in the hybrid imported fire ant colony is derived from S. invicta.

Thesis (Ph. D.)--University of Missouri-Columbia, 2003. / Typescript. Vita. Includes bibliographical references (leaves 185-199). Also available on the Internet.

Thesis (Ph. D.)--University of Missouri-Columbia, 2003. / Typescript. Vita. Includes bibliographical references (leaves 185-199). Also available on the Internet.