Anopheles é o gênero da família Culicidae mais estudado devido sua importância médica. Atualmente o gênero Anopheles compreende 472 espécies válidas que estão divididas em sete subgêneros. Os principais vetores de plasmódio da Malária no Brasil pertencem ao subgênero Nyssorhynchus, que inclui 39 espécies oficialmente reconhecidas e um número crescente de complexos de espécies crípticas que estão distribuídas em três Seções: Myzorhynchella, Albimanus e Argyritarsis. Atualmente a Seção Myzorhynchella é formada por seis espécies: An. lutzii, An. parvus, An. nigritarsis, An. guarani, An. antunesi e An. pristinus. Para o desenvolvimento da análise morfológica, observou-se material-tipo depositado em diferentes coleções, espécimes depositados na coleção entomológica da FSP/USP, além de outros obtidos em coletas realizadas durante o presente estudo em diferentes localidades do Brasil. As análises moleculares foram desenvolvidas a partir de espécimes obtidos nas coletas. Revisão taxonômica da Seção Myzorhynchella é apresentada, incluindo-se descrições de quatro novas espécies e redescrições das demais, informações sobre bionomia, importância médica, caracterização molecular, distribuição geográfica, estado de preservação do material-tipo, além de chaves de identificação de adultos, larva de quarto estádio e genitália masculina. Os resultados das análises filogenéticas utilizando sequências de ITS2, COI e Catalase indicam a existência de pelo menos doze espécies dentro da Seção Myzorhynchella, os espécimes que vêm sendo identificados como An. antunesi constitui um complexo formado por possíveis cinco espécies e aqueles de An. parvus e An. pristinus também podem representar complexos de espécies. As sequências de ITS2 podem ser utilizadas como marcador diagnóstico para espécies da Seção Myzorhynchella. Contudo, o estudo ainda demonstra que pouco se conhece sobre a diversidade de espécies de Anopheles que ocorrem em ambientes onde a malária ocorre em baixa endemicidade. Pelo número de espécies novas encontradas e pela escassez de trabalhos com espécies da Seção, fica evidente a necessidade de mais estudos. / Anopheles is the Culicidae Family genus most studied because of its medical importance. The genus currently comprises 472 valid species that are divided into seven subgenus. The main vectors of plasmodium malaria in Brazil belong to the subgenus Nyssorhynchus, that includes 39 valid species and a growing number of cryptic species complexes that are divided in three Sections: Myzorhynchella, Albimanus and Argyritarsis. Myzorhynchella Section currently consists of six species: An. lutzii, An. parvus, An. nigritarsis, An. guarani, An. antunesi and An. pristinus. For the morphological analysis, type material deposited in different collections were studied, specimens deposited in the entomological collection of FSP/USP, and other obtained from field collections in different localities in Brazil during this study. The molecular analyzes were taken from specimens obtained in field collections. Taxonomic review of Myzorhynchella Section is presented, including descriptions of four new species and redescriptions of the others, information on bionomics, medical importance, molecular characterization, geographical distribution, preservation status of type material, as well as identification keys of adults, fourth instar larvae and male genitalia. The results of phylogenetic analysis using sequences of ITS2, COI and Catalase indicate the existence of at least twelve species within Myzorhynchella Section, the specimens which have been identified as An. antunesi is a species complex formed by possible five species and those of An. parvus and An. pristinus may also represent species complexes. The ITS2 sequences can be used as a diagnostic marker for species of Myzorhynchella Section. However, the study also shows that little is known about the diversity of Anopheles species that occur in environments where malaria occurs in low endemicity. By the number of new species found and the lack of studies with species of the Section, it is evident the need for further studies.

Non-melanoma skin cancer (NMSC) is the most common cancer in the world with a lifetime risk for development as high as 2 in 3 in Queensland, Australia. Mortality is quite low, representing an approximate 360 deaths in Australia annually but cost of treatment is extremely high, estimated at $232 million each year. Squamous cell carcinoma (SCC) and basal cell carcinoma (BCC) are the two most common forms of NMSC. Although BCC generally do not have the propensity to metastasise, they are highly invasive and can be locally destructive. SCC on the other hand is invasive and has metastatic potential. SCC is generally derived from a precursor lesion, solar keratosis (SK), which is also considered to be a biomarker of BCC, SCC and malignant melanoma. According to one theory, SKs actually represent the first recognisable stage of SCC development and therefore may be indicative of the earliest stage of NMSC development. In addition to these common forms of NMSC, rarer forms such as keratoacanthoma (KA), which spontaneously regress, and SCC in situ, which rarely become invasive, may provide clues into protective mechanisms associated with prevention of development. Like all other cancers, NMSC arises from an accumulation of genetic abnormalities that result in severe cellular dysfunction. A number of genes have been proposed in the development of NMSC, including p53, CDKN2a, Bcl-2 and the Ras family of genes, which are typically associated with proliferative and differentiation processes. Also, a number of genetic disorders that predispose individuals to NMSC have also been identified. Genetic abnormalities in these genes may be a result of somatic mutations that may be promoted by environmental carcinogens. For NMSC, ultraviolet (UV) radiation is the primary environmental stimulus that acts upon skin to generate mutations. UV effects are 2-fold; the first being direct damage produced by UVB radiation and the second being indirect damage as a result of UVA-induced oxidative stress. In addition to mutations of genes that directly result in carcinogenesis, polymorphic variants of genes may also play a role in susceptibility to NMSC. These susceptibility genes may have immunogenic, detoxifying or transcriptional roles that could be involved in increased mutagenesis or activation of cancer causing genes. The purpose of this study was ultimately to identify further molecular based mechanisms associated with the development of non-melanoma skin cancer. Initially, this study aimed to examine the effects of aberrant chromosomal regions on NMSC development and also to identify candidate genes within these regions that may be implicated in the development and progression of NMSC. Also, based on chromosomal and functional implications, a number of candidate genes were assessed using association analysis to determine their involvement in susceptibility to the earliest stages of NMSC development. Implicated susceptibility genes were then further investigated to determine their response to UV radiation. Therefore the methodological approach of these studies was based on three broad technical applications of cytogenetic, association and expression analyses. Previous comparative genomic hybridisation (CGH) studies implicated the 18q chromosomal region in progression of SK to SCC and this region was therefore suspected of harbouring one or more tumour suppressor genes that were associated with a more malignant phenotype. Following on from this analysis, loss of heterozygosity (LOH) analysis was used for further delineation of this region and possibly to implicate candidate genes involved in progression. Additionally, CGH was used to investigate keratoacanthoma to determine aberrant regions that might be involved in progression and also regression of this NMSC. Genes that had potential functional roles in NMSC development and that were located in or near regions implicated by these cytogenetic analyses were further investigated using association analysis. Association analysis was performed using polymerase chain reaction and subsequent restriction enzyme digestion or GeneScan analysis to determine genotype and allele frequencies in an SK affected versus control population for polymorphisms within a number of candidate genes. This population was carefully phenotyped so that not only genotypic factors could be analysed but also their interaction with a number of phenotypic and environmental risk factors. Genes with polymorphisms that did show association with solar keratosis development were then examined functionally. Specifically, gene expression analysis was undertaken to investigate their response to UV radiation. Both UVA only and combined UVA/UVB treatments were used for short term irradiation and also for long term irradiation with recovery to determine differential effects of UV range and dose in human skin. Relative mRNA expression analysis of these genes was performed using quantitative real time reverse transcription polymerase chain reaction to determine if UV radiation imposed gene expression changes in the skin. A combination of these methodologies provided a wide basis for investigation of NMSC. Cytogenetic, association and expression analyses all allow for the identification of molecular risk factors that cause or are associated with NMSC development and progression. These analyses provided diverse results that implicated various molecular mechanisms in the development of NMSC. Cytogenetic analysis is a powerful technique, especially for the identification of a broad range of aberrations throughout the genome. This study employed LOH analysis to investigate an implicated region involved in progression to SCC and to attempt identification of candidate genes that may be involved in this process. LOH analysis was successfully performed on 9 SCCs, 5 SCCs in situ and 2 SKs using 8 microsatellite markers within the 18q region. Polymerase chain reaction (PCR) was used to amplify polymorphic regions of these markers and genotypic composition was determined for normal and cancerous tissue within the specimen. In heterozygote individuals, determined by analysis of normal tissue, the cancerous tissue was examined to determine if alleles within the implicated region had been lost. However, after analysis of multiple different samples, there was no LOH detected in any of the samples examined for this analysis. This does not necessarily reject a role for 18q, or genes within this region, as the localisation of candidate tumour suppressor genes within a small region may indicate a tighter region of involvement than was expected. As such, a more targeted study may further delineate this region and implicate candidate genes in progression of SK to the more malignant phenotype of SCC. Further CGH analysis of keratoacanthoma was also undertaken to identify aberrations associated with development and also regression of this skin cancer. CGH was performed using universal amplification and nick translation to incorporate a fluorescent dye. Differentially labelled normal and tumour DNA were then competitively hybridised to a normal metaphase spread and fluorescence emission indicated either amplification or deletion of specific chromosomal regions. In total, 6 KA samples were analysed, with 2 samples each from evolving, matured and regressing stages of KA development. In general, regressing KAs appeared to be more highly associated with deleted regions than evolving and matured KAs. Specifically, the 15q chromosomal region that was deleted in regressing KAs but amplified in evolving or matured KAs, may be significantly involved in the process of KA regression. Also various candidate genes that were being considered for analysis were located in or near some of these implicated regions, including GSTM1, GSTP1 and SSTR2. As such, these candidate genes were targeted for further investigation. A number of susceptibility genes that were located in or near aberrant regions implicated in NMSC development were investigated using association analysis. These genes included members of the somatostatin receptor family (SSTR1 and SSTR2), members of the glutathione-S-transferase (GST) family (GSTM1, GSTT1, GSTP1 and GSTZ1) and the vitamin D receptor (VDR). Studies detected a number of interesting interactions between genetic, environmental and phenotypic factors in the development of the early stages of non-melanoma skin cancer. Additionally, genes implicated in NMSC development were further investigated using expression analysis to determine response to UV radiation. Association analysis was initially performed on members of the somatostatin receptor family. Somatostatin is a growth inhibiting factor, amongst other things, that mediates its actions through the somatostatin receptors (SSTRs). The presence of these receptors (SSTR1-5) in tumour cells indicates a potential for somatostatin to bind and suppress growth, as well as allowing for therapeutic treatment with somatostatin analogues. Additionally, expression of these receptors in normal tissue, including skin, should allow for potential protection against tumour growth. The genes for SSTR1 and SSTR2 have been shown to contain dinucleotide repeat polymorphisms, and although these polymorphisms may not directly result in altered expression or binding potential, they may be linked to another functional polymorphism that does. Using association analysis the SSTR1 and SSTR2 genes were investigated to determine whether they play a role in the development of solar keratosis. Results showed that there were no significant differences between SSTR1 and SSTR2 polymorphism frequencies in the tested solar keratosis population (P = 0.10 and P = 0.883, respectively) as compared to an unaffected population. Hence, these studies do not support a role for the SSTR1 or SSTR2 genes in solar keratosis development. Further association analysis and subsequent expression analysis was also performed on members of the glutathione-S-transferase family. The GST enzymes play a role in the detoxification of a number of carcinogens and mutagens, including those produced by UV-induced oxidative stress. This study examined the role of GSTM1, GSTT1, GSTP1 and GSTZ1 gene polymorphisms in susceptibility to SK development. Association analysis was performed to detect allele and genotype frequency differences in SK affected and control populations using PCR and restriction enzyme digestion. No significant differences were detected in GSTP1 and GSTZ1 allele or genotype frequencies, however polymorphisms within both genes were found to be in linkage disequilibrium, as previously reported, and a new allelic variant of the GSTZ1 gene was identified. Significant associations between GSTM1 (P = 0.003) and GSTT1 (P = 0.039) genotypes and SK development were detected, with the null variants of both genes conferring an approximate 2-fold increase in risk for solar keratosis development (OR: 2.1; CI: 1.3-3.5 and OR: 2.3; CI: 1.0-5.0 for GSTM1 and GSTT1, respectively). For the GSTM1 gene, this risk was significantly higher in conjunction with high outdoor exposure (OR: 3.4; CI: 1.9-6.3) and although the GSTT1 gene showed a similar trend (OR: 2.9; CI: 1.1-7.7), this did not reach significance. The increased risk of SK development associated with these genes is likely due to a decreased ability of the skin to detoxify mutagenic compounds produced by UV-induced oxidative stress, and hence a decreased ability to protect against carcinogenesis. Implication of the GSTM1 and GSTT1 null variants in solar keratosis development prompted interest in analysis of gene expression changes in response to UV radiation. Due to the high homology of the GSTM1 gene with other GSTM genes, and therefore potential issues with primer specificity, the GSTT1 gene was focussed on for the expression studies. Real time reverse transcription PCR, incorporating SYBR green fluorescence and 18S as a comparative gene, was used to study GSTT1 gene expression changes in response to both UVA and combined UVA/UVB radiation. It was found that only short term UV radiation had an effect on GSTT1 expression changes, whereas no alteration of gene expression was seen after 4 and 12 hours of recovery from long term irradiation between irradiated and matched non-irradiated skin samples. This indicated that changes in gene expression for the GSTT1 gene apparently occur relatively quickly after exposure to UV radiation. Analysis of both UVA only and combined UVA/UVB short term irradiation indicated that an initial decrease in expression, followed by an increase was likely to represent translation into protein and subsequent transcription of mRNA, and in some cases a second decrease indicated further translation. Hence, it appears as though UV radiation does have a significant effect on the expression of at least one GST gene, and that UV radiation in combination with genetic variation of these genes may play a role in the development of NMSC. Finally, association and subsequent expression analysis was also performed on the vitamin D receptor. The hormonal form of vitamin D, 1a25 dihydroxyvitamin D3, has been shown to have numerous cancer-related effects, including antiproliferative, differentiation, proapoptotic and antiangiogenic effects. These effects are mediated through the binding of 1a25 dihydroxyvitamin D3 to the vitamin D receptor and subsequent transcriptional pathways. Polymorphisms within the VDR are known to regulate its transcription and therefore expression, which is linked to the ability of 1a25 dihydroxyvitamin D3 to bind. Association analysis of a 5’ initiation codon variant (Fok I) and two 3’ variants (Apa I and Taq I) was performed in SK affected and control populations. Although the Fok I variant showed no association with SK development, both the Apa I and Taq I variants were found to be associated with SK development (P = 0.043 and P = 0.012, respectively). In particular, the Aa and Tt genotypes were associated with increased risk of SK. These results were however more complicated, as shown by further analysis. This showed that genotypes containing at least one allele that conferred decreased VDR transcription (ie. AA/Aa and Tt/tt) increased risk of SK development by 2-fold in fair skinned individuals (OR: 2.1; CI: 1.2-3.7 and OR: 1.7; CI: 1.1-2.7 for Apa I and Taq I variants, respectively) but also found to decrease the risk of SK development by 2-fold in medium skinned individuals (OR: 0.5; CI: 0.3-1.0 for Apa I variants). Additionally, genotypes containing 2 alleles conferring decreased transcription of the VDR gene were found to further increase the risk for SK development in fair skinned individuals (OR: 2.5; CI: 1.4-4.5 and OR: 2.4; CI: 1.2-5.0 for Apa I and Taq I variants, respectively), indicating a possible additive effect for the alleles. The highly differential association of the VDR gene polymorphisms amongst phenotypes may reflect a combination between the ability of an individual to synthesise 1a25 dihydroxyvitamin D3 with the binding availability of the VDR. To further investigate the role of VDR in NMSC, expression analysis of the VDR gene was undertaken using real time reverse transcription PCR, with SYBR green fluorescence and 18S as a comparative gene, to examine expression pattern changes associated with UV radiation. It was found that short term irradiation, as well as long term irradiation and recovery were associated with gene expression changes. Short term irradiation resulted in patterns indicative of translation and subsequent transcription, whereas long term irradiated samples resulted in reduction of VDR expression that was recovered after an extended period of time. Thus, VDR expression is clearly influenced by UV exposure. It would be very interesting to see more specifically if particular VDR genotypes, which appear to play a role in NMSC risk, also are affected differentially by UV exposure. It is possible that VDR expression is reduced to limit excessive binding of 1a25 dihydroxyvitamin D3, although since both UVA and UVB radiation affect VDR expression, this may not be mediated the effect of 1a25 dihydroxyvitamin D3 but rather a different pathway resulting from a general UV response. In summary, the detection of a number of susceptibility genes involved in SK development and their subsequent expression analysis in response to UV radiation has given further insight into the molecular changes associated with NMSC. In fact, both detoxification genes (GSTM1 and GSTT1) and a transcription related gene (VDR), were found to confer susceptibility to solar keratosis, an early stage skin lesion with tumourigenic potential. This suggests that even the earliest stages of skin cancer are mediated through a wide range of effects. Additionally, expression changes related to these genes indicate that they are associated with the well known environmental carcinogen of UV radiation and that their effects may be mediated through a wide range of pathways. Although implication of the 18q region in SCC progression was not confirmed in this study, it is still likely to play a role in malignant transformation. The implication of this region, as well as the implication of susceptibility genes has vastly increased knowledge into processes associated with NMSC. Although additional analysis can confirm and further implicate these molecular alterations, this study has resulted in a more comprehensive understanding of NMSC that may ultimately be of benefit in terms of prognosis and treatment.

Η ανθρώπινη εντερική χλωρίδα είναι ένα «ουσιώδες» όργανο, το οποίο συμβάλλει στη θρέψη, την ανοσία, την ανάπτυξη των εντερικών επιθηλιακών κυττάρων του ξενιστή, αλλά και συμμετέχει σε διάφορα μεταβολικά μονοπάτια αυτού. Επηρεάζεται από ποικίλους παράγοντες, όπως η ηλικία, ο γονότυπος του ξενιστή, το περιβάλλον, το στρες, η δίαιτα, καθώς και η πρόσληψη προβιοτικών, πρεβιοτικών, αντιβιοτικών. Η ανθρώπινη εντερική χλωρίδα αποτελείται κυρίως από δύο φύλα βακτηρίων: τα Bacteroidetes και τα Firmicutes. Τα τελευταία χρόνια πολλοί ερευνητές έχουν επικεντρώσει το ενδιαφέρον τους στη μελέτη της σχέσης που έχει η παχυσαρκία με την εντερική χλωρίδα. Τα αποτελέσματα είναι αντικρουόμενα. Συγκεκριμένα, έχει αποδειχθεί ότι σε παχύσαρκα άτομα υπάρχει χαμηλότερο ποσοστό Bacteroidetes και μεγαλύτερο Firmicutes, σε σχέση με άτομα φυσιολογικού βάρους. Στη συγκεκριμένη μελέτη ερευνήθηκε η επιρροή της παχυσαρκίας, της απώλειας βάρους, της διατροφής και της μεσογειακής δίαιτας στη σύσταση της εντερικής χλωρίδας. Η πειραματική πορεία που ακολουθήθηκε είναι η εξής: απομόνωση DNA, ηλεκτροφόρηση, φωτομέτρηση και Real Time – PCR. Η παχυσαρκία και ο αυξημένος Δείκτης Μάζας Σώματος σχετίστηκε, σε στατιστικά σημαντικό βαθμό, με μειωμένα ποσοστά Bacteroides και Bifidobacterium στα κόπρανα. Ακόμα, η απώλεια βάρους σχετίστηκε με μειωμένη ποσότητα Lactobacillus. Η υιοθέτηση της Μεσογειακής Διατροφής δε φάνηκε να σχετίζεται με διαφορές στην εντερική χλωρίδα σε στατιστικά σημαντικό βαθμό. Ενώ, όσον αφορά τη διατροφική πρόσληψη, βρέθηκε η κατανάλωση φρέσκων φρούτων, μοσχαριού και αναψυκτικών να σχετίζεται αρνητικά με την ποσότητα Clostridium. Η κατανάλωση οσπρίων σχετίστηκε θετικά με την ποσότητα Bacteroides, ενώ την ποσότητα των Bifidobacterium φάνηκε να επηρεάζει αρνητικά η κατανάλωση φρέσκων φρούτων και θετικά η κατανάλωση λαχανικών. / Gut flora is an «essential» part of human life that contributes positive to nutrition, to immunity and to the growth of the epithelial gut cells of the host. Moreover, it sprigthfully participates at host’s different metabolic paths. Gut flora is affected by various factors, such as the age, host’s genotype, environment, stress, diet and intake of probiotics, prebiotics or antibiotics. The gut flora of human consists of two main bacterial genders: The Bacteroidetes and the Firmicutes. Last years, many scientists have focused their interest on researches which deal with the correlation between the obesity and the gut flora. But, there are conflicting results on these researches. In particular, it is proved that the humans who suffer from obesity have smaller percentage of Bacteroidetes and bigger percentage of Firmicutes concerning with humans with normal weight. This research tries to correlate the obesity, the weight loss, the nutrition and the Mediterranean diet with the gut flora. The research has the next experimental development: DNA isolation, electrophoresis, photometry and Real Time – PCR. Obesity and increased Body Mass Index were related, in significant level, with reductions in Bacteroides and Bifidobacterium percentage in feces. Additionally, weight loss was related with decreased Lactobacillus percentage in feces. Adherence to Mediterranean Diet was not related with significant changes in gut microbiota. The consuming of fresh fruits, beef and soft drinks was, in significant level, related negatively with Clostridium levels in feces. The consuming of legumes was related positively with Bacteroides levels and, finally, Bifidobacterium levels in feces were affected negatively by fresh fruits consuming and positively by salads consuming.

O gênero Ramalina Ach. possui cerca de 250 espécies aceitas e está distribuído por todo o mundo. Desde a circunscrição inicial do gênero em 1809, aceitava-se que boa parte das espécies possuía distintos graus de plasticidade morfológica e até química, resultando em centenas de nomes na literatura e certa confusão a respeito da sistemática do mesmo. Estudos recentes revelaram uma diversidade oculta para o grupo e que, possivelmente, algumas características estavam sendo negligenciadas. Com o objetivo de contribuir para o entendimento da sistemática de Ramalina, o presente estudo utilizou-se de análises morfológicas, anatômicas, químicas e moleculares. No total foram examinados 2415 espécimes de Ramalina, dos quais 332 representam espécimes tipo. Foram geradas 278 novas sequências relativas a 24 espécies, sendo 195 sequências de ITS, 59 de IGS e 24 de rpb2. O estudo identificou 27 espécies distribuídas por doze estados brasileiros e em diversos ambientes, tais como restingas, mata de araucária, campos de altitude, Cerrado e Caatinga. Duas espécies são novas para a ciência. Ramalina anceps foi cofirmada como espécie distinta de R. usnea. A espécie R. subfraxinea foi excluída da micota brasileira. Foram registradas 33 novas ocorrências em vários Estados brasileiros, o que amplia consideravelmente a distribuição das espécies no país. A importância de ter descrições detalhadas sobre a morfologia e anatomia foi comprovada através dos estudos tendo respaldo das análises moleculares. O presente estudo contribui muito para o conhecimento a cerca dos problemas taxonômicos e sistemáticos das espécies de Ramalina, não apenas em nível nacional, mas mundial. Sobe para 38 o número de espécies de Ramalina conhecidas para o Brasil. A Região Sul teve substancial ganho no conhecimento a respeito das espécies presentes, bem como parte da Região Sudeste e Nordeste. As Regiões Centro-Oeste e Norte ainda carecem de coleções suficientes mesmo em herbários nacionais. Descrições, comentários, ilustrações e chaves de identificação são apresentadas. / Ramalina Ach. has ca. 250 species accepted and is distributed worldwide. Since the initial division of the genus in 1809, it was accepted that many of the species had different degrees of morphological and even chemical plasticity, resulting in hundreds of names in literature and some confusion about the systematic of the group. Recent studies have revealed a hidden diversity to the group and possibly some characteristics were being neglected. In aim to contribute to the understanding of systematic of Ramalina, this study used morphological, anatomical, chemical and molecular analyzes. 2415 specimens of Ramalina were examined, of which 332 represent type specimens. 278 new sequences related to 24 species were generated: 195 sequences of ITS, 59 of IGS and 24 of rpb2. The study identified 27 species distributed on twelve Brazilian states and in different environments, such as coastal vegetation, Araucaria forest, high altitude grasslands, savannah and Caatinga vegetation. Two species are new to science. Ramalina anceps was confirmed as a distinct species of R. usnea. The species R. subfraxinea was excluded from the Brazilian mycota. Thirty three new records were found in some Brazilian states, which considerably expand the distribution of species in the country. The importance of having detailed descriptions of morphology and anatomy has been proven through the studies and supported by the molecular analysis. This study contributes to the knowledge about the taxonomic and systematic problems of the species of Ramalina, not only nationally, but worldwide. The number of Ramalina species known to Brazil increases to 38. The South region had substantial gain to the knowledge about the species present, and part of the Southeast and Northeast regions. The North and Midwest regions still lack of sufficient collections even in national herbaria. Descriptions, comments, illustrations and identification keys are given.

Submitted by Náthali Maria Machado de Lima (nathalimachadolima@gmail.com) on 2018-09-06T17:50:08Z No. of bitstreams: 1 Nathali Maria Machado de Lima - Copia (2)-pages-1-3,5-101-merged.pdf: 3526956 bytes, checksum: 5f5c8cc704c14910caa7d290299d4c24 (MD5) / Approved for entry into archive by Elza Mitiko Sato null (elzasato@ibilce.unesp.br) on 2018-09-06T18:17:51Z (GMT) No. of bitstreams: 1 lima_nmm_me_sjrp.pdf: 3526956 bytes, checksum: 5f5c8cc704c14910caa7d290299d4c24 (MD5) / Made available in DSpace on 2018-09-06T18:17:51Z (GMT). No. of bitstreams: 1 lima_nmm_me_sjrp.pdf: 3526956 bytes, checksum: 5f5c8cc704c14910caa7d290299d4c24 (MD5) Previous issue date: 2016-10-31 / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Crostas biológicas consistem de uma comunidade composta por cianobactérias, algas verdes, microfungos, bactérias, líquens e musgos. Estas ocorrem em uma variedade de solos e regiões climáticas ao redor do globo, exercendo funções importantes como a de conferir estabilidade e proteger o solo contra forças erosivas, acumular e aumentar o tempo de residência da água no solo, além de, favorecer a germinação e permitir a fixação de nitrogênio e carbono. Em crostas biológicas de regiões quentes e temperadas há o predomínio de líquens e cianobactérias, sendo que as cianobactérias são consideradas os primeiros organismos a estruturarem a crosta, sendo seguidas por outros grupos de organismos. A partir da investigação de cianobactérias em crostas biológicas, muitos novos gêneros e espécies têm sido descritos, o que também enfatiza a grande necessidade da investigação sobre tais comunidades. Devido às condições propícias do ambiente, em termos de exposição do solo e fitofisionomia, o bioma Cerrado (Savana) foi escolhido para ser o local de estudo de assembleias de cianobactérias de crostas. Além disso, devido a estudos prévios que relatam o potencial de produção de enzimas celulolíticas e hemicelulolíticas por cianobactérias heterocitadas e a presença comum deste tipo de organismos em crostas biológicas, também foram feitas análises para avaliar a produção de tais substâncias. Desse modo, os estudos objetivaram contribuir com conhecimento da biodiversidade de cianobactérias de crostas biológicas de solo e avaliar a capacidade de produção enzimática das cianobactérias encontradas em crostas biológicas de solo. Para isto, amostras de crostas foram coletadas nos Parques Nacionais da Serra da Canastra (MG) e da Serra do Cipó (MG), no Parque Estadual Furnas do Bom Jesus (SP) e na região de Caldas Novas (GO). As cianobactérias encontradas nas amostras foram estudadas morfologicamente a partir do material da natureza e também isoladas em cultivos artificiais, visando além do estudo morfológico, as análises moleculares e enzimáticas. No total, foram estudados 28 morfotipos, provenientes de 31 populações, dos quais 12 foram identificados em nível específico (destes, três ainda exigem confirmação de identidade = cf) e 16 foram identificados apenas em nível genérico. Dezenove populações foram analisadas também em nível molecular por meio do sequenciamento parcial do gene 16S rRNA, e completo da Região Espaçadora Interna Transcrita 16S-23S (ITS). Seis populações foram também investigadas quanto a produção de enzimas. Entre os gêneros encontrados, muitos são comuns em assembleias de cianobactérias de crostas biológicas em outras regiões do mundo, entretanto, algumas identificações em nível de espécie não foram possíveis por diferenças importantes com os táxons já descritos em literatura. As análises moleculares enfatizaram e contribuíram com a problemática polifilia de gêneros como Leptolyngbya, Nostoc e Calothrix, reafirmaram a existência de um novo gênero próximo a Wilmottia e Microcoleus e apresentaram um novo grupo composto até o momento por uma espécie representada por duas populações. Este grupo revelou sequências moleculares que se aproximam de Brasilonema, entretanto os indivíduos apresentam ramificações verdadeiras e, portanto, investigações mais aprofundadas são necessárias para definição da identidade das populações. Dessa forma, as análises morfológicas e moleculares demonstraram a grande diversidade não acessada em localidades e habitats pobremente investigados e enfatizaram a contribuição deste trabalho ao enfocar pela primeira vez a flora de cianobactérias de solo de Cerrado. As análises de atividade enzimática revelaram que não houve produção de tais proteínas, tendo como possíveis explicações a ausência de habilidade para produção deste tipo de enzimas por parte das cepas, ou a ineficiência do método utilizado. De qualquer forma os resultados enfatizaram a necessidade de estudo nesta área, principalmente pela dificuldade no encontro de cepas produtoras e o desconhecimento sobre quais poderiam ser os fatores a estarem influenciando certas cepas a ativarem ou selecionarem diferentemente seus metabolismos. Em linhas gerais, os resultados do presente estudo apresentam, pela primeira vez, a composição das cianobactérias de crostas biológicas do território brasileiro e indicam uma flora diversificada e, em grande parte, desconhecida. Esses resultados fornecem subsídios e abrem caminho para outras pesquisas dentro do campo do conhecimento da biodiversidade presente em crostas biológicas. Complementarmente, estes trabalhos compõem a base para estudos sobre a ecologia de ambientes mais restritivos (como a caatinga ou o próprio Cerrado), permitindo abordagens como sucessão ecológica, produtividade primária, fluxos de nutrientes e dinâmicas de solo. / Biological soil crusts (BSCs) are communities potentially composed of cyanobacteria, green algae, micro fungi, bacteria, lichens and mosses. They occur in a variety of soils and climatic regions around the world, playing important roles as giving soil stability protecting against erosive forces, accumulating and increasing residence time for water in soil, besides promoting germination and performing nitrogen and carbon fixation. Biological soil crusts from hot deserts are frequently composed of cyanobacteria and lichens, and the cyanobacteria are considered the first colonizers in structuring the crust, being followed by other groups of organisms. A lot of new genera and species have been described based on crusts investigations and this fact also emphasizes the necessity of works on those communities. Due to environmental conditions, as soil exposition and phytophysiognomy, the biome Cerrado (Savannah) was chosen to be the place for studies on cyanobacterial assemblages of biological soil crusts. Besides, due to previous studies that indicate the production capability of celulolitic and hemicelulolitic enzymes by heterocytous cyanobacteria and the common presence of this kind of cyanobacteria in biological soil crusts, analyses of such production were also conducted. In this way, the studies aimed to contribute with knowledge about cyanobacterial biodiversity in biocrusts and evaluate the bioprospection capability of cyanobacterial from these biocrusts. Therefore, BSCs were sampled at the National Parks of Serra da Canastra (Minas Gerais State) and Serra do Cipó (Minas Gerais State), at the State Park of Furnas do Bom Jesus (São Paulo State) and in the region of the municipality of Caldas Novas (Goiás State). The cyanobacteria found in the samples had their morphology analyzed from the natural and cultivated conditions. Besides, molecular and enzymatic analysis were carried out. The results summarized 28 morphotypes from 31 populations, which 12 were identified at specie level (three of them need identity confirmation = cf) and 16 were identified only at genus level. Nineteen populations also were studied with molecular methods, through partial sequencing of 16S rRNA gene and complete sequencing of the Internal Transcribed Space Region (16S-23S ITS). In addition, six populations also were explored as possible enzymatic producers. Among the found genera, many are cyanobacteria frequently found in BSCs distributed around the world, however, some identifications at specific level were not possible due to considerable divergences in comparison with described taxa. The molecular analysis reaffirmed and emphasized the polyphyletic nature of Leptolyngbya, Nostoc and Calothrix, reinforced the existence of a new genus close related with Wilmottia and Microcoleus and presented a new group composed, until now, of one species represented by two populations. This group showed molecular sequences related with Brasilonema, however, the specimens are true branched, what requires more detailed studies to confirm the identification of the populations. In this way, the morphological and molecular analyses showed the wide diversity whose has not been accessed in poorly investigated places and habitats and reinforced the contribution of this work in focusing the cyanobacterial flora of crusts of Brazilian savannah area for the first time. The enzymatic activity analysis revealed that the strains studied did not produce celulolitic or hemicelulolitic compounds, having as possible explanations the absence of the production ability or the inefficiency of the utilized method. Either way, the results emphasized the necessity of studies in this area, mainly because of the difficulty in find producer strains and the lack of knowledge about which could be the factors influencing the strains in activate or select differently their metabolism. In general, our results presented for the first time the cyanobacterial composition for BSCs from Brazil and indicated a diverse, and sometimes, unknown flora. These results provided foundation and opened doors to investigations inside the biodiversity knowledge with biological soil crusts. Complementary, these works compound the basis for investigations in the ecology of extreme environments (as Caatinga and Cerrado), permiting studies about successional ecology, primary productivity, flow of nutrients and soil dynamics. / 2014/06245-8

Introdução: Mucopolissacaridose IVA é uma doença lisossômica, autossômica recessiva, causada pela deficiência da enzima N-acetilgalactosamina-6-sulfatase. É uma doença rara e a incidência varia de 1:76.000 a 1:640.000 recém-nascidos vivos. Até o momento 319 diferentes mutações causadoras da doença já foram identificadas, o que demonstra a ampla variabilidade genética. Objetivo: Caracterizar o genótipo de pacientes com MPS IVA, analisar 6 polimorfismos intragênicos e identificar os haplótipos presentes em nossos pacientes, através do estudo molecular do gene GALNS. Métodos: O estudo foi realizado em 45 pacientes provenientes das regiões Nordeste, Sudeste, e Sul do Brasil, com diagnóstico bioquímico confirmado para MPS IVA. A análise molecular foi realizada através de PCR seguida de sequenciamento, pelo método de Sanger, a fim de identificar as mutações causadoras da doença. Para o estudo de haplótipos foram analisados 6 polimorfismos intragênicos através de PCR em Tempo Real, pelo método Taqman, em pacientes e controles. Resultados: A análise do gene GALNS, nos 45 pacientes, permitiu a identificação de 18 diferentes mutações, e a caracterização de 6 haplótipos distintos. Das 18 mutações encontradas, 5 apresentaram uma alta frequência (p.Ser341Arg, p.Arg386Cys, p.Gly301Cys, p.Arg94Leu e p.Gly116Ser), além disso, foram encontradas 4 novas mutações em outros três pacientes (p.Gly115Arg, p.Asn45Gly, p.Thr394Ala e c.759-2A>G). Dentre as mutações encontradas com maior frequência, a mutação p.Ser341Arg foi identificada em um maior número de pacientes, sendo a maioria proveniente da região Nordeste. Além disso, todos os pacientes com esta mutação apresentaram um único haplótipo. Conclusão: Os resultados obtidos permitiram a identificação de 18 mutações dentre elas 4 novas mutações. A alta frequência da mutação p.Ser341Arg no Nordeste do Brasil, principalmente no estado da Paraíba nos leva a inferir um possível efeito fundador da doença. Esta mutação foi observada somente na população brasileira e todos os pacientes com mutação em homozigose apresentaramum único haplótipo. Estas análises são importantes para identificar portadores nas famílias, para diagnóstico pré-natal, e também como forma de identificar uma origem comum em mutações frequentes em determinadas populações. / Background: Mucopolysaccharidosis IVA is an autosomal recessive lysosomal disease, caused by deficiency of N-acetilgalactosamina-6-sulfatase. It is a rare disease and the incidence ranges from 1: 76,000 to 1:640,000 live births. To date 319 mutations have been identified in this gene, demonstrating the wide variability of disease causing mutations. Objective: Analyze and characterize the genotype of patients with MPS IVA, through molecular analysis of GALNS. Methods: Molecular analysis of 45 patients with confirmed biochemical diagnosis for MPS IVA was performed. Mutation analysis was performed by PCR followed by Sanger sequencing. Haplotype analysis was performed using 6 intragenic polymorphisms by Real-Time PCR. Results: In this study we found 18 different mutations among 45 Brazilian patients and identified 5 common mutations (p.Ser341Arg, p.Arg386Cys, p.Gly301Cys, p.Arg94Leu e p.Gly116Ser). Four novel mutations were also identified through molecular analysis, including: p.Gly115Arg, p.Asn45Gly, p.Thr394Ala e c.759-2A>G. Patients are distributed in Northeast, Southeast and South regions of Brazil. Six different haplotypes were identified among patients. The p.Ser341Arg mutation showed the highest frequency, and most patients are located in the Northeast, additionally, all patients with this mutation show the same haplotype.Conclusion: These analyzes are important to identify carriers in families, for prenatal diagnosis, and in order to identify the mutation origin when certain recurrent mutation is associated with the same haplotype. In this study, we observed a high frequency of p.Ser341Arg mutation in Northeast, mainly in the state of Paraíba. This mutation was detected with higher frequency among patients, and showed only a haplotype. This mutation is unique for the Brazilian population and thus, we could suggest that a possible founder effect for this mutation could exist.

Introdução: Mucopolissacaridose IVA é uma doença lisossômica, autossômica recessiva, causada pela deficiência da enzima N-acetilgalactosamina-6-sulfatase. É uma doença rara e a incidência varia de 1:76.000 a 1:640.000 recém-nascidos vivos. Até o momento 319 diferentes mutações causadoras da doença já foram identificadas, o que demonstra a ampla variabilidade genética. Objetivo: Caracterizar o genótipo de pacientes com MPS IVA, analisar 6 polimorfismos intragênicos e identificar os haplótipos presentes em nossos pacientes, através do estudo molecular do gene GALNS. Métodos: O estudo foi realizado em 45 pacientes provenientes das regiões Nordeste, Sudeste, e Sul do Brasil, com diagnóstico bioquímico confirmado para MPS IVA. A análise molecular foi realizada através de PCR seguida de sequenciamento, pelo método de Sanger, a fim de identificar as mutações causadoras da doença. Para o estudo de haplótipos foram analisados 6 polimorfismos intragênicos através de PCR em Tempo Real, pelo método Taqman, em pacientes e controles. Resultados: A análise do gene GALNS, nos 45 pacientes, permitiu a identificação de 18 diferentes mutações, e a caracterização de 6 haplótipos distintos. Das 18 mutações encontradas, 5 apresentaram uma alta frequência (p.Ser341Arg, p.Arg386Cys, p.Gly301Cys, p.Arg94Leu e p.Gly116Ser), além disso, foram encontradas 4 novas mutações em outros três pacientes (p.Gly115Arg, p.Asn45Gly, p.Thr394Ala e c.759-2A>G). Dentre as mutações encontradas com maior frequência, a mutação p.Ser341Arg foi identificada em um maior número de pacientes, sendo a maioria proveniente da região Nordeste. Além disso, todos os pacientes com esta mutação apresentaram um único haplótipo. Conclusão: Os resultados obtidos permitiram a identificação de 18 mutações dentre elas 4 novas mutações. A alta frequência da mutação p.Ser341Arg no Nordeste do Brasil, principalmente no estado da Paraíba nos leva a inferir um possível efeito fundador da doença. Esta mutação foi observada somente na população brasileira e todos os pacientes com mutação em homozigose apresentaramum único haplótipo. Estas análises são importantes para identificar portadores nas famílias, para diagnóstico pré-natal, e também como forma de identificar uma origem comum em mutações frequentes em determinadas populações. / Background: Mucopolysaccharidosis IVA is an autosomal recessive lysosomal disease, caused by deficiency of N-acetilgalactosamina-6-sulfatase. It is a rare disease and the incidence ranges from 1: 76,000 to 1:640,000 live births. To date 319 mutations have been identified in this gene, demonstrating the wide variability of disease causing mutations. Objective: Analyze and characterize the genotype of patients with MPS IVA, through molecular analysis of GALNS. Methods: Molecular analysis of 45 patients with confirmed biochemical diagnosis for MPS IVA was performed. Mutation analysis was performed by PCR followed by Sanger sequencing. Haplotype analysis was performed using 6 intragenic polymorphisms by Real-Time PCR. Results: In this study we found 18 different mutations among 45 Brazilian patients and identified 5 common mutations (p.Ser341Arg, p.Arg386Cys, p.Gly301Cys, p.Arg94Leu e p.Gly116Ser). Four novel mutations were also identified through molecular analysis, including: p.Gly115Arg, p.Asn45Gly, p.Thr394Ala e c.759-2A>G. Patients are distributed in Northeast, Southeast and South regions of Brazil. Six different haplotypes were identified among patients. The p.Ser341Arg mutation showed the highest frequency, and most patients are located in the Northeast, additionally, all patients with this mutation show the same haplotype.Conclusion: These analyzes are important to identify carriers in families, for prenatal diagnosis, and in order to identify the mutation origin when certain recurrent mutation is associated with the same haplotype. In this study, we observed a high frequency of p.Ser341Arg mutation in Northeast, mainly in the state of Paraíba. This mutation was detected with higher frequency among patients, and showed only a haplotype. This mutation is unique for the Brazilian population and thus, we could suggest that a possible founder effect for this mutation could exist.

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No. of bitstreams: 2 Dissertação - Tharlley Rodrigo Eugenio Duarte - 2018.pdf: 3238895 bytes, checksum: 4372ea2b66d739dd3a82a02b57e2b24a (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2018-04-26 / Brazil is the largest arbovirus granary in the world and presents the largest endemic area of yellow fever (YF). The Ministry of Health reported in 1170 suspected cases of yellow fever, of which 847 are under investigation, 93 were discarded and 230 were confirmed, being in the states of Minas Gerais (201), Espírito Santo (25) and São Paulo (4). Of the total number of cases reported, 186 died, 104 of which remained in the investigation, 79 deaths were confirmed and 3 were discarded. The case fatality rate among confirmed cases was 34.3%. For YF there is no specific treatment, however, vaccination is effective being the only and best way of prevention. Precisely because of this factor and others involved the diagnosis of YF is not made in the health system except by a relevant suspicion. The problem is that in many cases viruses go unnoticed in cases of Dengue virus infection because of cross reactivity between members of the genus Flavívirus or because of non-specific symptoms. The present study analyzed 118 samples that were screened for suspected Dengue Virus infections (VDEN) for the year 2011 to 2013, but discarded for this virus because they gave negative results to the viral agent through serological and molecular tests in the municipality of Goiânia, Goiás. Samples were sent to the Virology Laboratory of the Federal University of Goiás Regional Jataí and analyzed by molecular methods such as RT-PCR and Nested-PCR followed by verification of the amplicon by agarose gel electrophoresis. Among the 118 negative samples for the DENV virus, three of the samples were positive for the Yellow Fever Virus (YVF) according to the production of amplicons of 253 bp of the NS5 region and confirmation of the identity of the amplicon by nucleotide sequencing. The sequences obtained were submitted to BLAST for identity confirmation. The translated sequence was analyzed by MEGA software version 7.0. For phylogenetic analysis, the best model was previously determined and then the tree was constructed with 53 sequences of all Yellow Fever viruses present in databases for a comparative analysis. The sequences found were compared to sequences from the VFA recorded in the Genbank database and were identified as referring to a portion of the NS5 nonstructural protein, position 216 to 296 of this protein, conferring 81 amino acids. The representative tree demonstrated that the sequences submitted were directly related to Senegal taxa. The robustness of the phylogenetic method was by Bootstrap 2000 replicates using the best JTT + G model, Maximum Likelihood. The Tajima test applied yielded a value of D = 1.159570, thus demonstrating that there was no population expansion of the taxa analyzed, considering that they have a significant degree of conservation during evolution. From the results obtained in the study, it can be affirmed that there was already an YFV circulation in the year 2013, at least of patients seen in the central region of Brazil, even before the last outbreak in 2017. / O Brasil é o maior celeiro de arbovírus do mundo e apresenta a maior área endêmica de febre amarela (FA). O Ministério da Saúde notificou, em 2017, 1170 casos suspeitos de febre amarela, sendo que desses 847 estão em investigação, 93 foram descartados e 230 foram confirmados, sendo nos estados de Minas Gerais (201), Espírito Santo (25) e São Paulo (4). Do total de casos notificados, 186 evoluíram para óbito, sendo que 104 óbitos permanecem em investigação, 79 óbitos foram confirmados e 3 foram descartados. A taxa de letalidade entre os casos confirmados foi de 34,3%. Para FA não há tratamento específico, no entanto, a vacinação é eficaz sendo a única e melhor maneira de prevenção. Justamente por esse fator e outros envolvidos o diagnóstico da FA não é feito no sistema de saúde a não ser por uma suspeita relevante. O problema é que em muitos casos os vírus passam despercebidos em casos de infecção pelo vírus Dengue por apresentar reatividade cruzada entre membros do gênero Flavívirus ou por apresentar sintomas inespecíficos. O presente estudo analisou 118 amostras que foram triadas para infecções suspeitas de Vírus da Dengue (VDEN) referentes ao ano de 2011 a 2013, porém descartadas para esta virose por terem dado resultados negativos para o agente viral através de testes sorológicos e moleculares no município de Goiânia, Goiás. As amostras foram encaminhadas para o Laboratório de Virologia da Universidade Federal de Goiás Regional Jataí e analisadas por métodos moleculares, como o de RT-PCR e Nested-PCR seguida da verificação do amplicon por eletroforese em gel de Agarose. Dentre as 118 amostras negativas para os vírus DENV, três das amostras foram positivas para o Vírus da Febre Amarela (VFA) conforme produção de amplicons de 253 pb da região NS5 e confirmação da identidade do amplicon por sequenciamento nucleotídico. As sequências obtidas foram submetidas ao BLAST para confirmação da identidade. A sequência traduzida foi analisada pelo software MEGA versão 7.0. Para análise filogenética foi determinado previamente o melhor modelo e em seguida a árvore foi construída com 53 sequências de todos os vírus da Febre Amarela presentes em bancos de dados para uma análise comparativa. As sequências encontradas foram comparadas com sequências do VFA registradas no banco de dados Genbank e identificouse que se refere a uma porção da proteína não estrutural NS5, posição 216 a 296 desta proteína, conferindo 81 aminoácidos. A árvore representativa demonstrou que as sequências submetidas estavam diretamente relacionadas com táxons do Senegal. A robustez do método filogenético foi por Bootstrap 2000 réplicas utilizando o melhor modelo JTT+G, Maximum Likelihood (Máxima probabilidade). O teste D de Tajima aplicado gerou um valor de D= 1.159570, demonstrando assim que não houve expansão populacional dos táxons analisados, considerando que os mesmos possuem significativo grau de conservação durante a evolução. A partir dos resultados obtidos no estudo, pode-se afirmar que já havia circulação do VFA no ano de 2013, pelo menos de pacientes atendidos na região central do Brasil, antes mesmo do último surto em 2017.

Introdução Anopheles (Nsssorhynchus) constitui o grupo de anofelinos que encerra o maior número de vetores de plasmódios que causam a malária humana na Região Neotropical. Em vista disso, são as espécies que têm sido mais frequentemente estudadas. O subgênero possui 33 espécies e está dividido em três seções, Myzorhynchella, Albimanus e Argyritarsis. A última revisão da seção Myzorhynchella é a de Galvão (1941) e são raros os estudos com a seção que é formada pelas espécies An. lutzii, An. parvus, An. nigritarsis e An. antunesi. Embora estas espécies sejam consideradas zoofílicas, estudos taxonômicos são necessários para estabelecer a identificação morfológica e para diferenciar estas espécies de outros Anophelinae, fornecendo assim condições adequadas para avaliar as espécies que estão envolvidas na transmissão da malária. Objetivos Caracterizar morfologicamente e molecularmente as espécies da seção Myzorhynchella e estabelecer caracteres morfológicos que permitam a separação entre as mesmas. Métodos Foram realizadas coletas de mosquitos em diferentes localidades da Mata Atlântica, além da análise de caracteres morfológicos de larva, pupa, adultos macho e fêmea e ovos de espécimes disponíveis na coleção entomológica da Faculdade de Saúde Pública FSP/USP, do Museu de Zoologia MZUSP e do Instituto Oswaldo Cruz IOC. Foram realizadas análises moleculares utilizando sequências de bases nucleotídicas da região do Espaçador Interno Transcrito 2 - ITS2 do DNA ribossômico e do gene mitocondrial Citocromo Oxidase Subunidade I - COI. Resultados - Foram caracterizados os adultos, machos e fêmeas, as formas imaturas e os ovos de An. antunesi e de An. lutzii. Anopheles guarani e An. niger foram retiradas da sinonímia de An. lutzii. Foi descrita uma espécie nova que é encontrada em simpatria com An. antunesi na Serra da Mantiqueira. Os resultados das análises filogenéticas corroboraram a existência de pelo menos cinco espécies dentro da seção Myzorhynchella e indicam que An. parvus e An. antunesi podem representar complexos de espécies. Acresce considerar que An. lutzii foi redescrita com o emprego de espécimes do Vale do Ribeira. No entanto, a falta de espécimes de An. lutzii da localidade tipo com as formas adultas e imaturos associados, impediram a caracterização adequada da espécie. Conclusão Foram caracterizadas quatro espécies da seção Myzorhynchella, foi descrita uma espécie nova que ocorre na Serra da Mantiqueira e demonstrou-se que An. parvus e An. antunesi podem ser complexos de espécies. Há a necessidade de continuar os estudos da Seção Myzorhynchella e obter topotipos de An. lutzii / Introduction Anopheles (Nsssorhynchus) is the group of anophelines that has the largest number of vectors of plasmodium that causes human malaria in the Neotropics. Because of this, species of this subgenus have been most frequently studied. The subgenus has 33 nominal species, subdivided into three sections, Myzorhynchella, Albimanus and Argyritarsis. The last revision on species of the Myzorhynchella section is that by Galvão (1941) and taxonomic studies are rare in the section that is formed by An. lutzii, An. parvus, An. nigritarsis and An. antunesi. Although these species are considered to be zoophilic, taxonomic studies are necessary to fix the morphological identification, and to differentiate these species from other Anophelinae, thus providing appropriate conditions to evaluate which species are involved in the transmission of malaria. Objetives To fix the morphological and molecular identification of species of the Myzorhynchella section and to define morphological characters to separate the species. Methods The mosquitoes were collected in different localities in the Mata Atlântica. These specimens were employed in analyses of morphological characters of eggs, larva, pupa, adult male and female, and were compared to specimens deposited in the Entomological Collection of Faculdade de Saúde Pública FSP/USP, Museu de Zoologia MZUSP and Instituto Oswaldo Cruz IOC. Molecular analysis were performed using sequences of of the internal transcribed spacer 2 ITS2 of ribosomal DNA and the mitochondrial cytochrome oxidase subunit I gene COI. Results We characterized the adults, male and female, immature forms and the eggs of An. antunesi and An. lutzii. Anopheles guarani and An. niger were removed from the synomym of An. lutzii. A new species that was found in sympatry with An. antunesi in Serra da Mantiqueira was described. Results of phylogenetic analysis corroborate the existence of at least five species within Myzorhynchella Section and indicate that An. parvus and An. antunesi may represent complexes of species. Besides An. lutzii was redescribed using specimens of Ribeira Valley. However, the lack of specimens of An. lutzii from the type locality with associated adults and immature forms prevented the proper characterization of the species. Conclusion We characterized four species of Myzorhynchella section, a new species that occurs in Serra da Mantiqueira was described, and it showed that An. parvus and An. antunesi may be species complex. There is a need to continue the studies of the section and get topotypes of An. lutzii

Este trabalho visa esclarecer a identidade taxonômica de duas espécies de briozoários reportadas para o litoral brasileiro como Amathia cf. vidovici e Amathia cf. crispa, baseando-se em análises morfológicas e moleculares, utilizando os genes COI e 16S, que haviam se mostrado eficientes na diferenciação genética de outras espécies do gênero Amathia. Colônias foram amostradas em diversas localidades ao longo da costa brasileira, de Fortaleza, CE, a Palhoça, SC, tendo sido obtidas 107 amostras de A. cf. vidovici e 18 de A. cf. crispa. Uma espécie nova foi detectada para cada grupo avaliado: Amathia sp. nov.1 e Amathia sp. nov.2. Amathia sp. nov.1 é oriunda somente de localidades situadas em Cabo Frio, RJ e Vitória, ES, sendo distinta de A. cf. vidovici pelas medidas de maiores valores das seguintes características: 1) comprimento do agrupamento de zooides, 2) comprimento do internódio do estolão e 3) ângulo do agrupamento de zooides. Amathia sp. nov.2 foi encontrada exclusivamente na Ilha do Mel, PR revelando-se distinta de A. cf. crispa devido a medidas de menores valores das características: 1) ângulo da espiral, 2) número de pares de zooides, 3) comprimento do agrupamento de zooides e 4) comprimento do estolão. As análises moleculares corroboraram os resultados da morfologia, que conjuntamente revelaram também uma estrutura geográfica para A. vidovici. Esta foi encontrada em todo litoral do Brasil (CE, PB, PE, AL, BA, RJ, SP, PR, SC) sendo as colônias amostradas na ecorregião Nordeste, as que tiveram os menores valores mensurados das características analisadas quando comparadas com as das demais ecorregiões (maiores medidas). Amathia vidovici havia sido reportada anteriormente para localidades nos estados do Paraná, São Paulo e Alagoas, e Amathia crispa apenas para São Paulo / This work aims to clarify the taxonomic identity of two species of bryozoans reported on the Brazilian coast as Amathia cf. vidovici and Amathia cf. crispa based on morphological and molecular analysis using the mitochondrial genes 16S rRNA and COI, which had proved useful in the genetic differentiation at the species level of other species of genus Amathia. Colonies were collected in different localities along the Brazilian coast, from Fortaleza, Ceará state, up to Palhoça, Santa Catarina state, including 107 samples of A. cf. vidovici and 18 of A. cf. crispa. A new species was detected for each evaluated group: Amathia sp. nov.1 e Amathia sp. nov.2. Amathia sp. nov.1 was found only on sites situated in Cabo Frio, state of Rio de Janeiro, and Vitória, state of Espírito Santo, being distinct of A. cf. vidovici by having: 1) longer clusters of zooids, 2) longer stolon internodes, and 3) spiral of zooid clusters describing a greater angle. Amathia sp. nov.2 was exclusively found in Ilha do Mel, state of Paraná; it is distinct from A. cf. crispa by the following: 1) spiral of zooid clusters describing a smaller angle, 2) fewer pairs of zooids per cluster, 3) shorter clusters of zooids, and 4) shorter stolon internodes. The molecular analyses corroborate the morphological results, both also revealing a geographical structure for A. vidovici. This specie was found throughout the coast of Brazil (CE, PB, PE, AL, BA, RJ, SP, PR, SC), the ones from the Northeast Ecoregion with the lower values of all measured characteristics when compared with those from other ecoregions. Amathia vidovici was reported previously to localities in the states of Paraná, São Paulo, and Alagoas, and Amathia crispa was reported only to São Paulo