As espécies de moscas-das-frutas da família Tephritidae são consideradas importantes pragas da fruticultura mundial por utilizarem vários frutos de valor comercial como substrato para desenvolvimento do seu estágio de larva. O gênero Anastrepha é endêmico do Continente Americano e compreende cerca de 200 espécies descritas, das quais 99 ocorrem no Brasil. Dentre essas espécies, destaca-se a espécie nominal Anastrepha fraterculus (sensu lato), que compreende um complexo de espécies crípticas, quatro das quais foram reconhecidas, até o momento, A. sp.1, A. sp.2 e A. sp.3 (Brasil) e A. sp.4 encontrada no Equador. O presente trabalho buscou a caracterização da variabilidade do espaçador ITS1 do DNA ribossômico de A. sp.1, A. sp.2 e A. sp.3 em amostras populacionais de diversas localidades do Brasil e a análise desse espaçador em amostras de outras regiões das Américas do Sul, Central e México. O ITS1 mostrou ser um marcador bastante útil para análises filogenéticas entre espécies próximas. Os fragmentos amplificados com os iniciadores 18SF e 5.8SR, construídos neste trabalho, continham cerca de 900 pb, não havendo diferenças significativas entre as amostras. Os fragmentos continham uma seqüência parcial do gene 18S, o ITS1 completo e o início do gene 5.8S. O ITS1 variou de 539 a 584 bases. Todos os exemplares apresentaram uma quantidade muito maior de base AT (entre 77,6 a 88,6%). O índice de similaridade entre as amostras variou de 88% a 99,8%. Não foram observadas diferenças significativas intraespecíficas entre as seqüências de amostras populacionais das entidades brasileiras do complexo. A análise filogenética feita pelo programa POY gerou muitas árvores igualmente parcimoniosas. A árvore de consenso estrito agrupou as amostras em clados distintos. O clado (grupo externo) formado por C. capitata e C. rosa foi colocado em um ramo distinto e no outro clado, todas as amostras de Anastrepha. O clado de A. fraterculus, separou as amostras do México e Guatemala das demais, que formaram outro clado. Dentro deste clado, foram formados dois subgrupos, um com as amostras do Peru e Equador e um com as amostras do Brasil e Argentina. Dentro desses clados houve separação em sub-grupos de amostras de cada região geográfica, sendo observada, com algumas exceções uma relação desses grupos com as divisões biogeográficas da entomofauna da América Latina. Desta forma, a análise do ITS1 corrobora resultados anteriores indicando que há claramente três entidades do complexo fraterculus no Brasil (A. sp.1, A. sp.2 e A. sp.3) e que diferentes morfotipos devem existir nas diferentes regiões da América Latina. / Fruit flies of Tephritidae family are important pest for the worldwide fruitculture because many species use commercial variety of fruits as host for the development of thier larval stage. Anastrepha is endemic of the American Continent and comprises about 200 described species, 90 of which occurs in Brazil. Among these species the most important is the nominal A. fraterculus, that, actually, comprises a complex of cryptic species, the so-called \"fraterculus complex\". Four species of the complex has been characterized, three of which occurs in Brazil (A.sp.1, A.sp.2, A. sp.3) and one in Ecuador (A. sp.4). In the present analyses the ITS1 spacer of the ribosomal genes was characterized by PCR and sequencing, in samples of A. fraterculus s.l. from several populations in Brazil and from different regions of South, Central America and Mexico. Amplyfication was carried out with primers designed in the present analyses, 18SF and 5.8SR, that generate fragments of 900 bp containing a partial sequence of the 18S gene, the complete ITS1 and the initial portion of the 5.8S gene. The length of the fragment did not varied significantly among the samples. The ITS1 size varied from 539 to 584 bp, and contained a very high content of AT, from 77.6 to 88.6 %, among samples. Similarity of the ITS1 sequences was high (from 88% to 99.8%) among the samples, and no signficant intraspecific differences were found in the sequences of samples from different brazilian populations. The POY software produced many parcimonious phylogenetic trees, and the strict consensus tree grouped the samples in distinct clades. First, there was a separation of the clade of Ceratitis species (outgroups) from the clade of Anastrepha. Within this clade A. obliqua was isolated from a large clade containg all the samples of A. fraterculus s.l.. Within A. fraterculus, one clade grouped the samples from Mexico and Guatemala, another clade the samples from Ecuador and Peru and a clade with the samples from Brazil and Argentina. The sample of Colombia was found as independent branch of the tree. In each of these clades there were subgroups of the different samples, and it was observed a relationship of these clades and subgroups to the biogeographic divisions of the entomofauna of Latin America with a few exceptions. The analysis of ITS1 clearly corroborate previous data indicating that there are three biological entities of the fraterculus complex in Brazil. It also indicates that different entities must exist in other regions of Latin America.

This work covers stages one through seven (of nine stages) of a classical biological control program for Mediterranean fruit fly (=medfly), Ceratitis capitata (Wiedemann). Major research objectives concentrate on stage five (exploration and collection of natural enemies), and stage seven (testing and selecting natural enemies for additional work). Coffee was collected monthly from three locations in Kenya from November 1997 through July 1999. Four species of tephritid flies and ten parasitoid species were recovered. Four guilds of parasitoids were recorded, and two egg-prepupal endoparasitoids, Fopius caudatus (Szépligeti) and F. ceratitivorus (Wharton), were discovered. The oviposition behavior of these two species is contrasted. Domination of this tropical parasitoid assemblage by koinobionts is discussed relative to the dominance of temperate fruit-infesting tephritid systems by idiobionts. Fruit handling procedures were examined for impact on overall percent emergence and specifically percent emergence of flies versus parasitoids. It was determined that stirring samples had a significant positive effect on overall emergence, however daily misting of fruit did not. The only treatment without a significant bias in fly emergence over parasitoids was the stirred/dry treatment. Effects of these results on rearing procedures are discussed. Host specificity and host suitability of parasitoids reared from coffee were examined via: (1) association of parasitoids with host flies based on characteristics of the fly puparia from which parasitoids emerged, (2) rearing of cucurbit infesting tephritids and their parasitoids in Kenya, (3) rearing of flowerhead infesting tephritids and their parasitoids in Kenya and Hawaii, and (4) host range testing of Psyttalia species in Kenya and Hawaii. These results are discussed in terms of their utility for predicting nontarget effects. Psyttalia concolor (Szépligeti) was shipped to Hawaii and tested against the nontarget gall forming tephritid Procecidochares utilis Stone introduced to control the weed Ageretina adenophora (Maui pamakani). Psyttalia concolor failed to attack the gall-forming P. utilis both in choice and no-choice tests, but readily attacked tephritid larvae offered in fruit in choice tests. Recommendations for further testing and release of the parasitoids from Kenya are discussed for Hawaii and Latin America.

Differential gene expression plays a key role in developmental pathways within an organism. Examples of such pathways include primary sex determination signaling and the formation of secondary sexual characteristics. This dissertation is focused on the use of suppression subtractive hybridization (SSH) to identify genes that are differentially expressed and involved in some aspect of sexual development in the Mediterranean fruit fly (medfly), Ceratitis capitata. In the course ofthis project, a method for sexing individual specimens from pre-adult stages was developed. This method was used to collect sex-specific RNAs at different developmental stages for use in SSH. A total of25 subtraction products were obtained across all the stages examined. Analysis of these products revealed that approximately half were similar to cytoplasmic ribosomal proteins and mitochondrial ribosomal RNA The remaining products represent putative medfly homologs of other previously identified genes or potentially novel genes One ofthe subtraction products, representing a potentially novel gene, was characterized in detail. This gene, named mapotge', represents a novel medfly gene that appears to encode a polypeptide of 299 amino acids. The N-terminus of this polypeptide contains a BTB-POZ domain. This domain functions as a protein-protein interaction motif found in a wide range of organisms from humans to Drosophila that mediates protein dimerization and oligomerization. The temporal expression pattern of mapotge' was determined using RT-PCR and Northern blot analysis. These revealed that the transcript is expressed throughout embryogenesis in both females and males, and in adult females that are > 0.5 days post-eclosion. Minimal expression is observed in female and male third instar larvae, early pupae, and in adult males. Studies were also initiated to characterize the representation of additioual sequences containing a BTB-POZ domain in the medfly genome. This was performed using Southern blot analysis and degenerate primers for the polymerase chain reaction (PCR). These results indicate the presence of at least three sequences in the medfly, in addition to 'mapotge', that contain a BTB-POZ domain. Potential evolutionary relationships ofthe BTB-POZ domain sequences from the medfly and other insect species were also analyzed.

Male Drosophila melanogaster double the transcription of most of the genes on their single X chromosome, to equal that from the two female X chromosomes, in a process termed dosage compensation. This process is mediated by the MSL complex, consisting of both protein and non-coding RNA components. This complex is only active in males due to the presence of MSL2, which is not translated in females. The X-linked Lsp1α gene of Drosophila melanogaster appears to escape dosage compensation, and exhibits two-fold higher levels of expression in females compared to males. The apparent lack of dosage compensation of Lsp1α could be due to the promoter being more active in females than in males, or to a lack of regulation by the MSL complex. In this study, the mechanism by which this happens has been addressed. Lsp1α is expressed exclusively in the fat body tissue of third instar larvae, and forms part of a multi-protein complex that acts as a nutrient reservoir during pupariation. In this study it has been shown that transgenes, in which the reporter gene, lacZ, is under the control of the Lsp1α promoter, exhibit variable levels of increased activity in female compared to male third instar larvae. At high levels of transgene expression, activity of the transgene is equal in female and male larvae. When the expression of the transgene is low, the activity of the transgene is much higher in female compared to male larvae. This increased sensitivity of the Lsp1α promoter to position effects in females appears to be mediated by one or more components of the MSL complex. Females ectopically expressing MSL2 exhibit decreased levels of transgene activity. Furthermore, overexpression of MSL1 causes an increase in the activity of transgenes subject to strong position effects. Despite these findings, the sex-specific regulation of the Lsp1α promoter does not account for the non-dosage compensated appearance of Lsp1α. Instead, unlike control dosage compensated X-linked genes, Lsp1α is not enriched for a histone modification, acetylation of lysine 16 of histone H4 that is essential for dosage compensation by the MSL complex. The developmental stage at which the four genes flanking Lsp1α are expressed has been determined using northern RNA hybridization. Expression of the gene immediately 3' of Lsp1α could not be detected at any developmental stage using northern RNA hybridization or in adults by RT-PCR. However, the two genes flanking Lsp1α are expressed in equal levels in male and female Drosophila as determined by quantitative RNase protection analysis. Furthermore, the regions between Lsp1α and these flanking dosage compensated genes do not prevent dosage compensation of an X-linked armlacZ reporter gene. Bioinformatic analysis shows that Lsp1α is present in three species closely related to D. melanogaster but is absent in more distantly related species. It is probable that because of its recent evolutionary origin, the Lsp1α gene lacks the DNA sequences that are required to attract the MSL complex. More generally, a model is proposed in which dosage compensation involves binding of the MSL complex to DNA sequences in actively transcribed regions with possible limited spreading to closely associated active genes.

Τα έντομα και άλλοι ποικιλόθερμοι οργανισμοί έχουν αναπτύξει διάφορους μοριακούς μηχανισμούς για την προστασία τους από ακραίες θερμοκρασίες και άλλες στρεσογόνες καταστάσεις που συμβαίνουν κατά τη διάρκεια της ζωής τους. Ένας από τους σημαντικότερους μηχανισμούς είναι η παραγωγή των θερμοεπαγόμενων πρωτεϊνών οι οποίες, ως μοριακοί συνοδοί, προστατεύουν τα κύτταρα από τη μετουσίωση και συσσωμάτωση των πρωτεϊνών τους. Πολλές από τις θερμοεπαγόμενες πρωτεΐνες συνθέτονται και σε φυσιολογικές συνθήκες και παίζουν σημαντικούς ρόλους στην ορθή αναδίπλωση, μεταφορά και αποικοδόμηση των πρωτεϊνών του κυττάρου. Το εργαστήριο μας ενδιαφέρεται για το ρόλο των θερμοεπαγόμενων πρωτεϊνών και τη ρύθμιση των γονιδίων τους στη Μεσογειακή μύγα. Τέσσερα θερμοεπαγόμενα γονίδια, hsp90, hsp70, hsp27 και hsp26, έχουν κλωνοποιηθεί και χαρακτηριστεί στο εργαστήριό μας. Στόχοι αυτής της μεταπτυχιακής εργασίας είναι α) η λειτουργική ανάλυση της 5’ ανοδικής περιοχής του γονιδίου hsp90 της Μεσογειακής μύγας και β) η έκφραση του γονιδίου σε συνθήκες ψυχρού στρες. Για να μελετήσουμε την ρύθμιση του γονιδίου hsp90, δημιουργήσαμε διαγονιδιακά στελέχη που έφεραν το γονίδιο της λουσιφεράσης (ως γονίδιο αναφοράς) υπό τον έλεγχο διαφορετικών 5’ περιοχών του γονιδίου hsp90. Η ανάλυση των στελεχών αυτών έδειξε ότι η ευρύτερη περιοχή του υποκινητή που εκτίνεται έως και το -3536 περιλαμβάνει όλα τα απαραίτητα στοιχεία για τη θερμοεπαγόμενη έκφραση του γονιδίου. Για να μελετήσουμε εάν η χαμηλή θερμοκρασία (0 oC) έχει κάποια επίδραση στη βιωσιμότητα του εντόμου και στην έκφραση του hsp90, ομάδες αρσενικών ενήλικων εντόμων επωάστηκαν για διαφόρους χρόνους στους 0 oC και ακολούθως μελετήθηκε η επιβίωσή τους και η έκφραση του θερμοεπαγόμενου γονιδίου με RT-PCR και qRT-PCR. Τα αποτελέσματά μας υποδηλώνουν ότι οι χαμηλές θερμοκρασίες αυξάνουν την έκφραση του hsp90 σχεδόν στα ίδια επίπεδα με εκείνα του θερμικού στρες. Επίσης η βιωσιμότητα των εντόμων βρέθηκε πολύ υψηλή στους 0 oC. / Insects and other poikilotherms have multiple molecular mechanisms for their protection from extreme environmental and other stressful conditions which happens during their life. One of these mechanisms is the production of heat shock proteins. These proteins, being molecular chaperones, protect the cells from protein denaturation and aggregation. Some of these proteins are produced under normal conditions, and they play important role in proper protein folding, transport and degradation. We are interested in the role of heat shock proteins and the regulation of their genes in Mediterranean fruit fly (medfly). Four heat shock genes have been cloned and characterized in our laboratory. These are hsp90, hsp70, hsp27 and hsp26. The aim of this master thesis is: a) to study the normal and heat-induced regulation of the medfly’s hsp90 gene and b) to study the expression of this gene under cold shock conditions. For the first aim we created transgenic flies which they had the lusiferase gene (as a reference gene) under the control of different 5’ upstream regions of hsp90 gene. The analysis of these transgenic strains showed that the region between the first transcription base pair and -3536 base pair is crucial for the expression of the gene under normal and heat shock conditions. In order to examine whether the low temperature (0 oC) has any effect in the medfly’s viability and the expression of hsp90 gene, male groups were exposed at different intervals at 0 oC and then it was examined the viability and the expression of the gene. Our results suggest that the low temperature increases the expression of hsp90 nearly to the same levels of the thermal stress and this temperature does not affect the viability of the insect.

A técnica do inseto estéril visa a indução de esterilidade em fêmeas selvagens por meio do acasalamento com machos estéreis liberados em grandes quantidades no ambiente. Os insetos liberados devem ser capazes de competir com os selvagens pela cópula com as fêmeas, fertilizar seus ovos e evitar que ela copule novamente. A recópula com machos selvagens pode interferir na eficiência da técnica por resultar em ovos férteis. Não é conhecido como a sequência de cópula-recópula com machos estéreis e/ou selvagens determina a fertilidade dos ovos na mosca-da-fruta, Ceratitis capitata (Wied., 1824) (Diptera: Tephritidae), nem as conseqüências deste comportamento, o que justificou o presente trabalho. Os experimentos foram realizados em laboratório, na Embrapa Semi-Árido, Petrolina-PE, sob condições ambientes de temperatura e umidade relativa, para determinar a influência do tipo e ordem das cópulas na indução de esterilidade, além de parâmetros do comportamento de recópula, como frequência recópula entre as fêmeas, número de cópulas for fêmea, tempo de cópula e recópula, período refratário e competitividade sexual dos machos. Foi também avaliada a interferência do tratamento aromático dos machos estéreis com óleo de gengibre sobre a recópula das fêmeas. A Biofábrica Moscamed Brasil, Juazeiro- BA forneceu os insetos estéreis, e os selvagens foram provenientes de pupas coletadas de frutos infestados em áreas frutícolas na região do Submédio do Vale do São Francisco. Na presença de machos estéreis e selvagens concorrendo pelas cópulas na proporção de 5 estéreis: 1 selvagem, 63% das fêmeas recopularam em média 3,37 vezes com curto período refratário entre a cópula e a primeira recópula. O tratamento dos machos com óleo de gengibre diminuiu significativamente a taxa de recópula. A aromaterapia dos machos não reduziu a fecundidade e nem conferiu aos machos vantagem da competição espermática, entretanto aumentou a indução de esterilidade em fêmeas selvagens que recopularam. O segundo macho a copular a fêmea apresentou maior vantagem reprodutiva em curto período refratário. Conclui-se que para que a técnica do inseto estéril seja eficiente, é necessário que haja sempre machos estéreis de boa qualidade no campo para recopularem as fêmeas em curto período refratário. / The sterile insect technique (SIT) aim at the induction of sterility in wild females by mating with sterile males released in great amounts in the field. These released males must compete with wild ones being able to attract and mate wild females, fertilize their eggs and avoid remating with a wild male that would lead to fertile eggs and consequently reduce the efficiency of the technique. The circumstances in which remating of Ceratitis capitata (Wied., 1824) (Diptera: Tephritidae) females interfere with the SIT, as well as the consequences of this behavior are not well known, what justified the present study. The experiments were carried out at the Embrapa Semi-Árido, Petrolina-PE, inside the laboratory under ambient conditions of temperature and relative humidity to determine the influence of the order of mating type in the induction of sterility. The parameters evaluated to better understand the remating behavior were remating frequency, number of matings per female, mating and remating time, refractory period, sexual competitiveness of males, and the interference of males aromatherapy with ginger root oil in the females remating. Moscamed facility, Juazeiro-BA, provided the sterile males and the wild ones were from pupae collected from infested fruits from commercial orchards in the São Francisco River Valley. In the presence of sterile and wild males in competition (5 sterile: 1wild), 63% of females remated in average 3.37 times in short refractory period between the mating and the first remating. The males aromatherapy with ginger root oil reduced significantly the remating rate, but did not reduce the fecundity, neither confer advantage to the males in the sperm competition, however increased the induction of sterility in wild females multiply mated. The second male to mate the female presented reproductive advantage if the refractory period was short. For the SIT be efficient the sterile males in the field must always have great quality to remate the females in a short refractory period and thus induce sterility in the wild population.

As espécies de moscas-das-frutas da família Tephritidae são consideradas importantes pragas da fruticultura mundial por utilizarem vários frutos de valor comercial como substrato para desenvolvimento do seu estágio de larva. O gênero Anastrepha é endêmico do Continente Americano e compreende cerca de 200 espécies descritas, das quais 99 ocorrem no Brasil. Dentre essas espécies, destaca-se a espécie nominal Anastrepha fraterculus (sensu lato), que compreende um complexo de espécies crípticas, quatro das quais foram reconhecidas, até o momento, A. sp.1, A. sp.2 e A. sp.3 (Brasil) e A. sp.4 encontrada no Equador. O presente trabalho buscou a caracterização da variabilidade do espaçador ITS1 do DNA ribossômico de A. sp.1, A. sp.2 e A. sp.3 em amostras populacionais de diversas localidades do Brasil e a análise desse espaçador em amostras de outras regiões das Américas do Sul, Central e México. O ITS1 mostrou ser um marcador bastante útil para análises filogenéticas entre espécies próximas. Os fragmentos amplificados com os iniciadores 18SF e 5.8SR, construídos neste trabalho, continham cerca de 900 pb, não havendo diferenças significativas entre as amostras. Os fragmentos continham uma seqüência parcial do gene 18S, o ITS1 completo e o início do gene 5.8S. O ITS1 variou de 539 a 584 bases. Todos os exemplares apresentaram uma quantidade muito maior de base AT (entre 77,6 a 88,6%). O índice de similaridade entre as amostras variou de 88% a 99,8%. Não foram observadas diferenças significativas intraespecíficas entre as seqüências de amostras populacionais das entidades brasileiras do complexo. A análise filogenética feita pelo programa POY gerou muitas árvores igualmente parcimoniosas. A árvore de consenso estrito agrupou as amostras em clados distintos. O clado (grupo externo) formado por C. capitata e C. rosa foi colocado em um ramo distinto e no outro clado, todas as amostras de Anastrepha. O clado de A. fraterculus, separou as amostras do México e Guatemala das demais, que formaram outro clado. Dentro deste clado, foram formados dois subgrupos, um com as amostras do Peru e Equador e um com as amostras do Brasil e Argentina. Dentro desses clados houve separação em sub-grupos de amostras de cada região geográfica, sendo observada, com algumas exceções uma relação desses grupos com as divisões biogeográficas da entomofauna da América Latina. Desta forma, a análise do ITS1 corrobora resultados anteriores indicando que há claramente três entidades do complexo fraterculus no Brasil (A. sp.1, A. sp.2 e A. sp.3) e que diferentes morfotipos devem existir nas diferentes regiões da América Latina. / Fruit flies of Tephritidae family are important pest for the worldwide fruitculture because many species use commercial variety of fruits as host for the development of thier larval stage. Anastrepha is endemic of the American Continent and comprises about 200 described species, 90 of which occurs in Brazil. Among these species the most important is the nominal A. fraterculus, that, actually, comprises a complex of cryptic species, the so-called \"fraterculus complex\". Four species of the complex has been characterized, three of which occurs in Brazil (A.sp.1, A.sp.2, A. sp.3) and one in Ecuador (A. sp.4). In the present analyses the ITS1 spacer of the ribosomal genes was characterized by PCR and sequencing, in samples of A. fraterculus s.l. from several populations in Brazil and from different regions of South, Central America and Mexico. Amplyfication was carried out with primers designed in the present analyses, 18SF and 5.8SR, that generate fragments of 900 bp containing a partial sequence of the 18S gene, the complete ITS1 and the initial portion of the 5.8S gene. The length of the fragment did not varied significantly among the samples. The ITS1 size varied from 539 to 584 bp, and contained a very high content of AT, from 77.6 to 88.6 %, among samples. Similarity of the ITS1 sequences was high (from 88% to 99.8%) among the samples, and no signficant intraspecific differences were found in the sequences of samples from different brazilian populations. The POY software produced many parcimonious phylogenetic trees, and the strict consensus tree grouped the samples in distinct clades. First, there was a separation of the clade of Ceratitis species (outgroups) from the clade of Anastrepha. Within this clade A. obliqua was isolated from a large clade containg all the samples of A. fraterculus s.l.. Within A. fraterculus, one clade grouped the samples from Mexico and Guatemala, another clade the samples from Ecuador and Peru and a clade with the samples from Brazil and Argentina. The sample of Colombia was found as independent branch of the tree. In each of these clades there were subgroups of the different samples, and it was observed a relationship of these clades and subgroups to the biogeographic divisions of the entomofauna of Latin America with a few exceptions. The analysis of ITS1 clearly corroborate previous data indicating that there are three biological entities of the fraterculus complex in Brazil. It also indicates that different entities must exist in other regions of Latin America.

Orientadores: Benedito Carlos Benedetti, Adalton Raga, Perla Gómez Di Marco / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia Agrícola / Made available in DSpace on 2018-08-17T18:23:01Z (GMT). No. of bitstreams: 1 Vieira_StellaMariaJanuaria_D.pdf: 942200 bytes, checksum: f0c6adbea884bee20c024b9044a05cb9 (MD5) Previous issue date: 2011 / Resumo: A goiaba é um fruto com boas características organolépticas e nutricionais, além de ter boa aceitação tanto para o consumo "in natura" como industrial. É, porém, bastante perecível após a colheita, daí decorrendo perdas significativas. Além disso, acaba destinada quase totalmente ao mercado interno, em virtude das barreiras fitossanitárias impostas por países importadores diante de problemas como a infestação por moscas-das-frutas. É exatamente no estudo dessa praga que se situa o objeto do presente trabalho, cuja realização obedeceu aos seguintes propósitos: 1) avaliar a eficiência dos métodos de controle de radiação "UVC"e "água quente", bem como a aplicação combinada de ambos, para o fim de promover a desinfestação de ovos de Ceratitis capitata (Wied.) (Diptera: Tephritidae) em goiabas 'Kumagai'; 2) avaliar a qualidade físico-química de goiabas 'Kumagai' submetidas aos tratamentos quarentenários. No que se refere ao método "água quente", o equipamento banho-maria Dubnoff existente em laboratório forneceu uma variação de temperatura de 0,5 ºC, sendo que as médias dos tempos de exposição, verificados em termopares inseridos a 5 mm, para as temperaturas avaliadas de 42,0 a 50,0 ºC (± 0,5) com intervalos de 1,0 ºC, foram menores que as médias dos tempos de exposição daqueles termopares inseridos no centro das goiabas para as mesmas temperaturas avaliadas. Em função desse aumento no tempo de imersão em altas temperaturas, foram observadas a presença de escaldaduras na casca das goiabas tratadas a 46,0; 47,0; 48,0; 49,0 e 50,0 ºC (± 0,5). Por sua vez, no que tange ao método por "UV-C", as lâmpadas utilizadas no equipamento de radiação possuíam comprimentos de onda predominantes de 254 nm (adequados, portanto, para o estudo de radiação UV-C) e a tela de aço (diâmetro da malha hexagonal: 6,0x 7,0 cm) foi o material escolhido para compor a prateleira central, distante 8 cm da fonte luminosa. Aplicados os métodos em questão, o que se observou nos testes "in vitro" foi que ampliação do tempo de imersão no tratamento hidrotérmico e a exposição a intensidades crescentes de radiação UV-C no tratamento por radiação conduziram a um aumento na mortalidade ovos de C. capitata; já quanto aos testes "in vivo", a conclusão, em relação ao método "água quente", foi a de que tratamentos a temperaturas de 46,0; 47,0; 48,0; 49,0 e 50,0 (± 0,5) ºC, com tempos de imersão de 0; 46; 36; 26; 16 e 12 minutos, respectivamente, são eficientes para a mortalidade de ovos de C. capitata (não tendo sido verificadas escaldaduras nas cascas das frutas às temperaturas avaliadas) e, em relação ao método "UV-C", verificou-se que a intensidade de 16,0 kJm-2 foi capaz de inativar 100% de ovos de C. capitata. Nos tratamentos combinados, foi observado efeito aditivo na mortalidade de ovos de C. capitata, sendo mais intensos nas combinações de 0,54 kJm-2 + 45,0 ± 0,5 ºC; 0,80 kJm-2 + 45,0 ± 0,5 ºC; 0,54 kJm-2 + 46,0 ± 0,5 ºC e 0,80 kJm-2 + 46,0 ± 0,5 ºC. Por fim, quanto à qualidade pós-colheita, a das goiabas submetidas aos tratamentos e armazenadas a 8,0 ± 0,2 ºC durante o período de armazenamento de 10 dias mostrou-se superior à das goiabas que tiveram aplicados os mesmos tratamentos e foram armazenadas a 22,0 ± 0,2 ºC / Abstract: Guava is a fruit that has good organoleptic and nutritional haracteristics, besides having a good acceptance as for consumption "in natura" as industrial. It is, however, a lot perishable after the harvest, and because of that there are countless losses to the producers. Besides it, the fruit is practically consumed in the internal market, because of the sanitary obstacles imposed by importing countries before matters as infestation by fruit flies. It is exactly in the study of this plague that is the object of this work whose achievement has obeyed the following steps: 1) evaluating the efficiency of hot water imersion, UV-C radiation methods, and the application in both, with the intention of promoting the not infestation of eggs of Ceratitis capitata (Wied.) (Diptera: Tephrittidae) in "Kumagai" guavas; 2) evaluating the physics-chemistry quality submitted to quarantine treatments. Mentioning to the method hot water, the double boiler Duboff equipment existing in the laboratory gives us a variation of temperature of 0,5 ºC, as for the average of times of expositions verified in pairs at 5 mm, for temperatures of 42,0 the 50,0 ºC (± 0,5) with intervals of 1,0 ºC was lower than the ones of the times in exposition in the center of guavas to the same temperatures. Due to this increase of the time of immersion in high temperatures, it was noticed the presence of burns on the treated guavas peel to 46,0; 47,0; 48,0; 49,0 and 50,0 ºC (± 0,5). By its shift, according to the UV-C method, the light bulbs used in the radiation equipment had 254 nm waves (proper to the study of radiation UV-C) and the steel screen (hexagonal mesh: 6,0 x 7,0 cm) was the chosen material to compose the central shelf, far 8 cm from the illuminated fountain. The used methods, what was noticed in the tests "in vitro" was that the amplification of the time of the immersion (in the hydrothermical) and the exposition to the increasing intensity of radiation UV-C (in the treatment by radiation) led to an increase in the mortality of eggs of C. capitata; according to the tests "in vivo" the conclusion in relation to the method hot water was that treatments to the temperatures 46,0; 47,0; 48,0; 49,0 e 50,0 ºC (± 0,5). With times of immersion of 46; 36; 26; 16 and 12 minutes, respectively, are efficient to the mortality of the eggs of C. capitata (not having being verified burns on the fruits peel to the evaluated temperatures) and, in relation to the method UV-C, it was noticed that the intensity of 16.0 kJm-2 was able to activate 100% of eggs of C. capitata. In the combined treatments, it was observed the adding effect in the mortality of eggs of C. capitata, being more intense in the combinations 0,54 kJm-2 + 45,0 ± 0,5 ºC; 0,80 kJm-2 + 45,0 ± 0,5 ºC; 0,54 kJm-2 + 46,0 ± 0,5 ºC e 0,80 kJm-2 + 46,0 ± 0,5 ºC. Finally according to the quality after the harvest, the guavas that were submitted to the treatment and stored to 8,0 ± 0,2 ºC during a period of 10 days were superior to the guavas that had the same treatments and were stored to 22,0 ± 0,2 ºC / Doutorado / Tecnologia Pós-Colheita / Doutor em Engenharia Agrícola

A assimetria de cinco estruturas corporais da mosca-das-frutas Ceratitis capitata foi analisada para uma avaliação se poderiam ser utilizadas como bioindicadores. As análises foram feitas em amostras de uma população natural e de uma população de laboratório. Adicionalmente, em amostras obtidas da população de laboratório, os índices de assimetria foram avaliados após a submissão das diferentes amostras à diferentes temperaturas. No estágio de pupa, as amostras foram tratadas nas temperaturas de 17, 20, 25 e 30oC. As estruturas analisadas, em machos e fêmeas foram as cerdas frontais (FO), orbitais (OB), pós-oculares (PO), o comprimento e largura das asas. Nos machos, além dessas estruturas, foi incluída a análise do comprimento de uma das cerdas orbitais, a supra-fronto-orbital (SFO), que apresenta dimorfismo sexual. A variação numérica das cerdas FO e OB foi muito baixa em todas as amostras, não permitindo uma análise da assimetria. A assimetria das demais características, tanto merísticas como métricas, mostrou ser compatível com o modelo da assimetria flutuante (AF). O grau de assimetria de cada estrutura não diferiu entre machos e fêmeas, mas foi significativamente mais alto na amostra da população de laboratório do que na amostra da população natural. No entanto, a assimetria holística (somatória da AF das diferentes estruturas) não mostrou diferenças entre as duas amostras. Nas amostras submetidas a diferentes temperaturas foram observadas alterações significativas no grau de assimetria das estruturas, mas nenhuma alteração no tipo de assimetria que continuou sendo caracterizada como flutuante. Essas análises mostraram não haver diferenças entre os sexos, nem interações entre sexo e temperatura, mas apenas diferenças significativas entre as temperaturas. A assimetria flutuante das cerdas pós-oculares aumenta com o elevação da temperatura, a AF do comprimento das asas e da cerda SFO (nos machos) não mostra correlação com as temperaturas e os resultados indicam que a AF da largura das asas é mais elevada nas temperaturas extremas que nas intermediárias. A comparação da assimetria holística entre as amostras das diferentes temperaturas mostrou que as diferenças não foram significativas. Os resultados indicam que a assimetria das 52 cerdas pós-oculares e das medidas do largura das asas apresentam potencial para evidenciar eventuais estresses durante o desenvolvimento desses insetos. Indicam, também, que a utilização de uma assimetria holísitca pode mascarar possíveis diferenças da assimetria flutuante de estruturas individuais. / Asymmetry of five traits of the fruit fly Ceratitis capitata, an introduced insect pest in Brazil, was analysed in order to evaluate if they present potencial to be used as bioindicators of the \"quality\" of these insects. Samples from a natural population and from a laboratory colony were studied. Moreover, asymmetry was measured in samples from the laboratory colony maintained during the pupal stage, in the temperatures of 17, 20, 25 and 30oC. The analysed traits of males and females, were the frontal, the orbital and the postocular bristles, measurement of the length and cross diameter of the wings and the length of the supra-frontoorbital bristles in the males that show a pronounced sexual dimorphism. Since the numerical variation of the frontal and orbital bristles was very low in all samples it was not possible to analyse the variation in terms of asymmetry. For all other traits, meristic or metric, asymmetry was characterized as following the fluctuating asymmetry (AF) model. AF of the different traits does not differ between sexes, but was significantly higher in the laboratory sample than in the sample from the natural population. No significant differences, however, were found in the holistic asymmetry (sum of AF of different traits) between the two samples. In relation to the experiments conducted in different temperatures, no variations in AF levels were observed between sexes, nor interactions between sex and temperatures. However, significant differences were observed among the samples submitted to different temperatures. AF of the postocular bristles increases proportionaly to the increment of the temperature, while AF of wings and supra-fronto-orbital bristles (in males) lengths showed no correlations with the temperatures, and AF of the cross diameter of wings seems to be higher at the extreme temperatures than at the intermediate ones. When a holistic AF was applied, no significant differences among the temperature samples were observed. The results indicate that asymmetry of the postocular bristles and the length of the wings are parameters showing potencial use as indicators of stresses during development of these insects. They also indicate that a holistic asymmetry may mask variations in the asymmetry of individual traits.

A família Tephritidae congrega várias espécies de moscas-das-frutas que utilizam frutos como substrato alimentar no estágio larval, adquirindo o status de inseto-praga quando esses frutos são de valor comercial. O gênero Anastrepha é endêmico do Continente Americano e compreende cerca de 212 espécies descritas, das quais 109 ocorrem no Brasil. A espécie nominal Anastrepha fraterculus representa um complexo de espécies crípticas e se encontra distribuída pela Região Neotropical e sul dos Estados Unidos. No Brasil, através do estudo de diversas características biológicas e do marcador molecular ITS-1 (espaçador ribossômico nuclear), identificou-se a existência de três espécies crípticas no complexo fraterculus, a Anastrepha sp.1 affinis fraterculus, A. sp.2 aff. fraterculus e A. sp.3 aff. fraterculus. Marcadores gênicos presentes no DNA mitocondrial, como o gene cytochrome oxidase I (COI), são ferramentas amplamente utilizadas em análises filogenéticas, pois esta molécula apresenta características distintas do DNA nuclear, como o fato de possuir herança predominantemente materna, apresentar ausência ou baixíssima taxa de recombinação na maioria dos táxons, além de altas taxas mutacionais. Estas características possibilitam a obtenção de dados importantes na interpretação das relações entre as espécies. Amostras do complexo fraterculus (A. sp.1, A. sp.2, A. sp.3) de 14 localidades (média de 5 indivíduos / localidade) no sudeste do Brasil, uma amostra de A sp.4 do Equador e dois grupos externos (A. grandis e A. striata) foram utilizados. Fragmentos de 1139bp do gene COI foram amplificados e sequenciados, 45 haplótipos foram identificados: 30 em A. sp.1, 5 em A. sp.2 e 17 em A. sp.3. A distância média entre as espécies foi de 0,021 e o Fst médio foi 0,347 indicando estruturação populacional muito alta e pequena distância entre os haplótipos, que não apresentaram diferenças fixadas entre as espécies. Os testes de desvio de neutralidade apresentaram valores significativamente negativos. Os testes de seleção evidenciaram a atuação de seleção purificadora com baixos valores de Ka/Ks e significância no Z-teste de seleção. A análise filogenética mostrou fortes evidências de introgressão e não separou as diferentes entidades em clados distintos. Houve a formação de dois ramos principais, um constituído quase que exclusivamente por amostras de A. sp.1, e apenas duas amostras de A. sp.3, e outro que reuniu todas as espécies do complexo. Os dois principais grupos de haplótipos também foram visualizados na rede de haplótipos que mostrou indícios de expansão populacional. Quando somado ao estudo sequências depositadas em bancos de dados por outros autores, a espécie nominal A. fraterculus apresentou em sua distribuição 5 grupos de haplótipos mitocondriais. Dois deles ocorrem no Brasil, um com amostras do México e Costa Rica, um na Guatemala e Venezuela (baixa latitude) e um com indivíduos da Colômbia e Venezuela (alta latitude), sendo que os grupos Brasileiros também reuniram amostras da Argentina e do Equador. Assim, as sequências de COI não permitem a caracterização das entidades do complexo fraterculus apesar de indicar a estruturação populacional e a hipótese mais provável é a de que tenha havido introgressão da molécula mitocondrial entre as espécies do complexo com posterior expansão / The Tephritidae family comprises fruit flies species whose larvae feed and develop in fruits, many of which are commercial varieties and thus the species assume economic significance. Anastrepha genus is distributed throughout the Neotropical region and Southern United States. Analyses of biological characteristics and of the internal transcribed spacer (ITS-1) of the nuclear ribosomal DNA allowed the characterization of three cryptic species of the fraterculus complex in Brazil: Anastrepha sp.1 affinis fraterculus, Anastrepha sp.2 aff. fraterculus and Anastrepha sp.3 aff. fraterculus. Mitochondrial markers as gene cytochrome oxidase I (COI) are largely used in phylogenetic analyses because they have maternal inheritance, none or low recombination and high mutation rates compared to the nuclear DNA. Hence, analyses of the complex based in this marker will offer a divergent perspective from nuclear DNA for inferences on the evolutive relationships between different species. Samples from the fraterculus complex (A. sp.1, A. sp.2, A. sp.3) from 15 localities (average of 5 individuals/ locality) in southeastern Brazil, one sample of A. sp.4 from Ecuador and two outgroups (A. grandis and A. striata) were employed and COI sequences of 1139bp were amplified and analyzed. We identified 45 haplotypes: 30 in A. sp.1, 5 in A.sp.2 and 17 in A. sp.3. The mean distance between the haplotypes was 0.021 and mean Fst 0.347, indicating high population structure and low mitochondrial distance. The neutrality tests had significantly neutral values. The selection tests revealed the action of purifying selection with low values of Ka/Ks and significance in the Z-test selection. Phylogenetic analysis showed strong evidences of introgression and did not separate the various entities in distinct clades grouping the three species in a single branch; there was also the formation of another main branch formed almost exclusively by strains of A. sp.1 and only two samples of A. sp.3. The two main groups of haplotypes were also seen in the haplotype network that showed evidence of population expansion. The analysis of the philogenetic tree based on mitochondrial COI showed strong evidence for introgression. No fixed differences between species were found though mtDNA marker shows a lot of polymorphism. When added sequences deposited in databases by other authors the nominal species A. fraterculus presented in its distribution five groups of mitochondrial haplotypes, two of them in Brazil, one with samples from Mexico and Costa Rica, one in Guatemala and Venezuela and one with individuals from Colombia. The Brazilian groups also collected samples from Argentina and Ecuador. Therefore, the COI sequences do not allow the characterization of the entities of the fraterculus complex, although structure among the species is shown. The most likely hypothesis is that introgression has happened in the mitochondrial molecule among the species with further expansion