Submitted by Claudia Rocha (claudia.rocha@udesc.br) on 2017-12-14T12:35:51Z No. of bitstreams: 1 PGEF17MA078.pdf: 124602 bytes, checksum: 743d0f80327ad1bfc6c95e6e3c31ebac (MD5) / Made available in DSpace on 2017-12-14T12:35:51Z (GMT). No. of bitstreams: 1 PGEF17MA078.pdf: 124602 bytes, checksum: 743d0f80327ad1bfc6c95e6e3c31ebac (MD5) Previous issue date: 2017-02-24 / Capes / The objective of this work was: (I) to characterize macroscopically and microscopically the anatomical structures of the wood of 31 species of Ombrophilous Mixed Forest, following the standardization of the International Association of Wood Anatomists (IAWA); (II) Identify its characteristics of taxonomic value; (III) Make identification keys using the microscopic characteristics of these species; (IV) To evaluate similarities and main anatomical components of the wood among 14 species. For the characterizations, the wood of species previously collected from the flooded areas for the implantation of the Hydroelectric Plant (HPP) of São Roque - Santa Catarina was used. Three specimens of each specimen were randomly selected from the specimens. One specimen was taken from each individual in the Diameter at Chest Height (DAP), after which two specimens were removed from the intermediate region of the disc. For the macroscopic characterization the wood was polished and classified according to the International Association of Wood Anatomists. In the microscopic description, two permanent slides and a macerated material were made for each test specimen. In the measurement of anatomical elements, the mean, minimum, maximum and standard deviation values were determined. Also, using the characterizations made, an identification key was made using the microscopic characteristics peculiar to each genus. Subsequently, of these 31 species, 14 were randomly selected to evaluate their similarities and their main anatomical components. In these 14 species, it was observed that the species present common characteristics such as diffuse porosity, vascular elements with simple perforation plate and stitches of the wall of the alternate vessels. In the Principal Components Analysis (PCA), the pore diameter, the length and thickness of the radius in micrometers, the length of the radius in number of cells, the vessel wall and the vascular wall were recorded / Este trabalho teve como objetivos: (I) Caracterizar macroscopicamente e microscopicamente as estruturas anatômicas da madeira de 31 espécies de Floresta Ombrófila Mista, seguindo a padronização da International Association of Wood Anatomists (IAWA); (II) Identificar suas características de valor taxonômico; (III) Confeccionar chaves de identificação utilizando as características microscópicas dessas espécies; (IV) Avaliar similaridades e componentes anatômicos principais da madeira entre 14 espécies. Para as caracterizações, foram utilizadas as madeiras de espécies previamente coletadas das áreas alagadas para a implantação da Hidrelétrica (UHE) de São Roque – Santa Catarina. Na obtenção dos corpos de prova foram selecionados aleatoriamente três indivíduos de cada espécie, sendo retirado um disco de cada indivíduo no Diâmetro a Altura do Peito (DAP), posteriormente foram retirados dois corpos de prova da região intermediária do disco. Para a caracterização macroscópica a madeira foi polida e classificada seguindo a norma da International Association of Wood Anatomists. Na descrição microscópica, para cada corpo de prova foram confeccionadas duas lâminas permanentes e um material macerado. Na mensuração dos elementos anatômicos determinou-se os valores médios, mínimos, máximos e o desvio padrão. Ainda, utilizando as caracterizações feitas, confeccionou-se uma chave de identificação utilizando as características microscópicas peculiares de cada gênero. Posteriormente, destas 31 espécies, foram selecionadas aleatoriamente 14 para avaliar suas similaridades e seus componentes anatômicos principais. Nestas 14 espécies, observou-se que as espécies apresentam características comuns como a porosidade difusa, elementos vasculares com placa de perfuração simples e pontoações da parede dos vasos alternas. Na Análise dos Componentes Principais (PCA), destacaram-se o diâmetro do poro, o comprimento e espessura do raio em micrômetros, o comprimento do raio em número de células, as pontoações da parede dos vasos e raio-vasculares

Content: Identification and classification of leather species becomes valuable and necessary due to concerns regarding consumer protection, product counterfeiting, and dispute settlement in the leather industry. Identification and classification of leather into species is carried out by histological examination or molecular analysis based on DNA. Manual method requires expertise, training and experience, and due to involvement of human judgment disputes are inevitable thus a need to automate the leather species identification. In the present investigation, an attempt has been made to automate leather species identification using machine learning techniques. A novel non-destructive leather species identification algorithm is proposed for the identification of cow, buffalo, goat and sheep leathers. Hair pore pattern was segmented efficiently using k-means clustering algorithm Significant features representing the unique characteristics of each species such as no.of hair pores, pore density, percent porosity, shape of the pores etc., were extracted. The generated features were used for training the Random forest classifier. Experimental results on the leather species image library database achieved an accuracy of 87 % using random forest as classifier, confirming the potentials of using the proposed system for automatic leather species classification. Take-Away: Novel technique to identify leather species Non destructive method Machine learning algorithms to automate leather species identification

Orientador: Nilton Nobuhiro Imai / Resumo: O objetivo desse doutorado é propor uma nova metodologia para identificar oito espécies arbóreas emergentes (i.e., que se sobressaem do dossel florestal), em diferentes idades e estágios de desenvolvimento e pertencentes à Mata Atlântica brasileira. Para tal, imagens hiperespectrais foram adquiridas em Julho/2017, em Junho/2018, e em Julho/2019 em um transecto localizado no fragmento florestal Ponte Branca, localizado a Oeste do Estado de São Paulo, onde a floresta é considerada estacional semidecidual e submontana. As imagens com resolução espacial de 10 cm foram adquiridas com câmara hiperespectral (500–900 nm) acoplada em veículo aéreo não tripulado (VANT ou UAV, do inglês Unmanned aerial vehicle) e, posteriormente corrigidas geometricamente e radiometricamente. Em seguida, as copas arbóreas individuais (ITCs, do inglês Individual tree crows) foram delineadas manualmente em cada conjunto de dados para serem utilizadas como referência para os experimentos. Dentre os experimentos realizados, destaca-se o uso do espectro normalizado para redução da variabilidade espectral intra-espécies, o uso da classificação baseada em regiões utilizando o algoritmo Random Forest e o uso de superpixexls para delineamento automático das ITCs em cada conjunto de imagens. Além disso, avaliou-se o uso dos superpixels multitemporais com diferentes atributos multitemporais (espectro normalizado, textura e índices de vegetação) e estruturais (derivados do modelo de altura das copas), sozinhos ou c... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The objective of this doctoral dissertation is to propose a new methodology to identify eight emergent tree species (i.e., that stood out from the canopy) belonging to highly diverse Brazilian Atlantic forest and with different ages and development stages. To achieve the objective, hyperspectral images were acquired in July/2017, June/208, and July/2019 in a transect area located in the western part of São Paulo State. The area is in Ponte Branca ecological station, where the forest is classified as submontane semideciduous seasonal with different stages of succession. Images with a spatial resolution of 10 cm were acquired with a hyperspectral camera (500–900 nm) onboard unmanned aerial vehicle (UAV) and geometrically and radiometrically post-processed. In sequence, the individual tree crowns (ITCs) were manually delineated in each dataset to be used as reference in the experiments. From the performed experiments, it is highlighted the use of mean normalized spectra to reduce the within-species spectral variability, the use of region-based classification with the Random Forest algorithm, and the use of superpixels to automatically delineate the ITCs in each dataset. Additionally, the multitemporal superpixels with different multitemporal features (normalized spectra, texture and vegetation indexes) and structural features derived from the canopy height model, combined or not, were assessed to the tree species classification. The best result was achieved merging normalized sp... (Complete abstract click electronic access below) / Doutor

While the presence of ≥1% of an undeclared species in ground meat generally used as an indicator of intentional mislabeling as opposed to cross-contamination, the actual percent of undeclared species resulting from cross-contamination has not been experimentally determined. The objective of this study was to quantify the effect of sanitation procedures on the crosscontamination of animal species in ground meat products, using undeclared pork in ground beef. Pork (13.6 kg) was processed using a commercial grinder, then one of three sanitation treatments was completed (“no cleaning”, “partial cleaning”, or “complete cleaning”). Next, beef (13.6 kg) was ground using the same equipment. For “no cleaning,” beef was ground immediately after pork without any cleaning step; for “partial cleaning,” the hopper tray was wiped, and excess meat was taken out from the auger; for “complete cleaning,” all parts of the grinder were disassembled and thoroughly cleaned with water and soap. A 100-g sample was collected for each 0.91 kg (2 lb) of beef processed with the grinder and each sanitation treatment was tested twice. Real-time polymerase chain reaction (PCR) was used to quantify pork in ground beef. For “no cleaning,” the first 100-g sample of ground beef run through the grinder contained 24.42 ± 10.41% pork, while subsequent samples contained

Mislabeling of ground meat products is a form of food fraud that can lead to economic deception and interfere with dietary restrictions related to allergens or religious beliefs. In various parts of the world, including Ireland, Mexico and Turkey, high levels of meat mislabeling have been reported between 2000-2015. However, there is currently a lack of information regarding this practice in the United States. Therefore, the objective of this study was to test a variety of ground meat products sold on the U.S. commercial market for the presence of potential mislabeling. Forty-eight ground meat samples were purchased from online and local retail sources, including both supermarkets and specialty meat retailers. DNA was extracted from each sample in duplicate and tested using DNA barcoding of the cytochrome c oxidase subunit I (COI) gene. The resulting sequences were identified at the species level using the Barcode of Life Database (BOLD). Any samples that failed DNA barcoding went through repeat extraction and sequencing. Due to the possibility of a species mixture, these samples were also tested with real-time polymerase chain reaction (PCR) targeting beef, chicken, lamb, turkey, pork and horse. Of the 48 products analyzed in this study, 10 were found to be mislabeled, with nine containing multiple meat species. Meat samples purchased from online specialty meat distributors had a higher rate of being mislabeled (35%) compared to samples purchased from a local butcher (18%) and samples purchased at local vii supermarkets (5.8%). Horsemeat, which is illegal to sell on the U.S. commercial market, was detected in two of the samples acquired from online specialty meat distributors. Overall, the mislabeling detected in this study appears to be due to reasons such as intentional mixing of lower-cost meat species into higher cost products or unintentional mixing of meat species due to cross-contamination during processing.

As abelhas sem ferrão estão entre os animais mais importantes para o equilíbrio do meio ambiente, isso devido a fatores como sua enorme diversidade e principalmente por serem importantes polinizadores tanto de ecossistemas naturais como de agroecossistemas. A necessidade de identificação dos animais amostrados por especialistas é fundamental para estudos ecológicos. Neste trabalho foram feitas análises interespecíficas e intra-populacionais sobre os indivíduos do gênero Schwarziana a fim de aperfeiçoar a utilização da técnica de morfometria geométrica, que nos permite identificações baseadas apenas nos padrões de venação das asas desses insetos. Não obstante, foram realizadas também análises de sequenciamento de dois genes do DNA mitocondrial, o COI e o 16S. O gênero estudado foi Schwarziana, com duas espécies válidas, S. quadripunctata e S. mourei. Nas análises morfométricas, os testes realizados por indivíduos os testes de validação cruzada identificaram de forma correta 70% das amostras de um total de 10 localidades diferentes. Esta acurácia aumenta ainda mais à medida que novos grupos são formados, alcançando próximo de 85% quando separadas por regiões. Em todos os ensaios realizados com a morfometria geométrica (partial warps e coordenadas alinhadas) atingiu-se uma taxa de 100% de identificação correta entre as duas espécies e nos ensaios feitos com as médias das colônias esta taxa também foi atingida para a identificação de todas as populações amostradas. Os testes feitos com os partial warps mostraram-se mais eficazes em relação às coordenadas alinhadas, já que nestes últimos a tolerância mínima para a separação dos grupos não foi atingida em alguns ensaios. As análises moleculares apontaram 53 sítios polimórficos para o gene COI, com um índice de diversidade nucleotídica de 0,02180 e de diversidade haplotípica de 0,8854, separando as amostras em 9 haplótipos, porém a rede de haplótipos não foi suficientemente conclusiva. A diferenciação genética total medida pelo parâmetro Fst somente para as populações de S. quadripunctata foi de 0.9453 (P<0,05). Já para o gene 16S foram encontrados 14 sítios polimórfico e um índice de diversidade nucleotídica de 0,00649 e de diversidade haplotípica de 0,8419, com 7 haplótipos gerados. O parâmetro Fst para todas amostras foi de 0.9552 (P<0,05) e somente para as de S. quadripunctata foi de 0.8736 (P<0,05). Para ambos os genes os testes Fst par-a-par mostraram uma maior variação entre as populações dos estados, mostrando que estas populações estão estruturadas. Os testes de Mantel correlacionaram positivamente os dados morfométricos, geográficos e moleculares. Das duas metodologias aplicadas em nossa pesquisa, podemos afirmar que a morfometria mostrou-se extremamente eficiente na diferenciação das populações amostradas. As análises moleculares indicaram que estas populações estão estruturadas mesmo analisando poucas amostras. No entanto ao unirmos estas duas metodologias, combinamos a simplicidade e a rapidez da morfometria geométrica com a capacidade sempre inovadora de estudos moleculares, obtendo desta forma ferramentas eficazes para acessar a biodiversidade em Meliponini, inclusive para rastrear geograficamente espécimes a partir de estudos com indivíduos de sua área de distibuição natural. / Stingless bees are among the most important animals to the environmental balance, that due to their diversity and mainly because they are important pollinators of both natural and agro-ecosystems. The need for identification of sampled animals by experts is essential for ecological studies. In this work, intra and interspecific population analysis was performed in order to improve the technique of geometric morphometry, which allows us to access this biodiversity through identifications based on patterns of wing venation of these insects. We also sequenced two mitochondrial genes, the COI and 16S. Schwarziana Lepeletier 1836 was the gender studied with two valid species, S. quadripunctata and S. mourei. In morphometric analysis, cross-validation tests carried out by individuals correctly identified 70% of samples from a total of 10 different locations. This accuracy increases further as new groups are formed, according to geographic proximity, reaching around 85% when separated by regions. In all tests with geometric morphometry (partial warps and aligned coordinates) reached a rate of 100% correct identification between the two species and the tests made with the averages of the colonies that rate achieved the perfect identification of all populations sampled. Partial warps tests were more effective in relation to aligned coordinates ones, as these last a minimum tolerance for the separation of the groups was not achieved in some tests. Molecular analysis showed 53 polymorphic sites for the COI gene, with nucleotide diversity of 0.02180 and haplotype diversity of 0.8854, separating the samples into 9 haplotypes, but their network of haplotypes was not sufficiently conclusive. The total genetic differentiation measured by Fst parameter only for the populations of S. quadripunctata was 0.9453 (P <0.05). As for the gene 16S, we found 14 polymorphic sites and a nucleotide diversity of 0.00649 and haplotype diversity of 0.8419, with seven haplotypes generated. The parameter Fst for all samples was 0.9552 (P <0.05) and only for S.quadripunctata was 0.8736 (P <0.05). For both genes Fst pairwise tests showed greater variation among populations of the states, showing these groups as a genetic structure. Mantel tests were positively correlated for morphometric, geographical and molecular data. Of the two methodologies in our research, we can affirm that the morphometry proved to be extremely efficient in the differentiation of populations. Molecular analysis showed that populations are consistent, even that a low sample size is available. However combining these two methodologies, we have joined the simplicity and speed of geometric morphometric with the always innovative ability of molecular studies, thus achieving effective tools for accessing biodiversity in Meliponini, including a geographically trace for specimens by studying individuals from their natural range.

Submitted by PPG Zoologia (zoologia-pg@pucrs.br) on 2018-05-18T17:22:44Z No. of bitstreams: 1 dissertacao_mestrado_final_paulochaves.pdf: 4426171 bytes, checksum: 8be6ef944f497d1a9518754ebfbc27c1 (MD5) / Approved for entry into archive by Sheila Dias (sheila.dias@pucrs.br) on 2018-05-28T12:19:27Z (GMT) No. of bitstreams: 1 dissertacao_mestrado_final_paulochaves.pdf: 4426171 bytes, checksum: 8be6ef944f497d1a9518754ebfbc27c1 (MD5) / Made available in DSpace on 2018-05-28T12:35:54Z (GMT). No. of bitstreams: 1 dissertacao_mestrado_final_paulochaves.pdf: 4426171 bytes, checksum: 8be6ef944f497d1a9518754ebfbc27c1 (MD5) Previous issue date: 2008-03-20 / Sequ?ncias de DNA usadas na identifica??o de material biol?gico t?m alcan?ado consider?vel popularidade nos ?ltimos anos, especialmente no contexto dos c?digos de barras de DNA. Aferir a esp?cie de origem em amostras de pelos, penas, peles e particularmente fezes ? um passo fundamental para quem estuda a ecologia e evolu??o de diversos animais com este tipo de amostra. Este ? o caso em carn?voros, cujos h?bitos furtivos e baixas densidades populacionais de algumas esp?cies evidenciam a import?ncia de estudos baseados em amostras n?o-invasivas. Entretanto a atual escassez de ensaios padronizados de identifica??o de carn?voros freq?entemente dificulta a aplica??o dessas amostras em larga escala e compara??es de resultados entre diferentes localidades. No presente estudo n?s avaliamos dois segmentos curtos (<250 pb) de DNA mitochondrial (mtDNA) localizados nos genes ATP sintase 6 e citocromo oxidase I com potencial de servirem como marcadores-padr?o para identifica??o de carn?voros. Entre um e 11 indiv?duos de 66 esp?cies de carn?voros foram seq?enciados para um ou ambos os segmentos do mtDNA e analisados usando tr?s diferentes m?todos (?rvore de dist?ncia, dist?ncia gen?tica e an?lise de caracteres). Em geral, indiv?duos conspec?ficos apresentaram menor dist?ncia gen?tica entre si do que em rela??o a outras esp?cies, formando agrupamentos monofil?ticos. Exce??es foram algumas esp?cies que divergiram recentemente, algumas das quais ainda puderam ser identificadas pelo m?todo de caracteres, hapl?tipos esp?cie-espec?ficos, ou reduzindo a abrang?ncia geogr?fica das compara??es (restringindo a an?lise a uma regi?o zoogeogr?fica). An?lises in silico, usando um segmento curto do citocromo b freq?entemente empregado em carn?voros, tamb?m foram realizadas para comparar o desempenho deste segmento em rela??o aos outros dois propostos. N?s ent?o testamos o desempenho destes segmentos na identifica??o de fezes de carn?voros por meio de tr?s estudos de caso: (i) fezes de felinos de zool?gico, objetivando-se verificar o potencial de contamina??o das seq?encias com DNA da presa (coelho); (ii) fezes coletadas no Cerrado brasileiro contendo restos de presas (p?los, ossos, penas), supostamente proveniente de lobo-guar?, objetivando-se investigar a efici?ncia de identifica??o do predador e ocorr?ncia de interfer?ncia do DNA da presa na identifica??o; e (iii) fezes coletadas em uma reserva na Mata Atl?ntica, tamb?m com o objetivo de avaliar a efici?ncia de identifica??o. Apesar de diferen?as em alguns aspectos de sua performance, nossos resultados indicam que os dois segmentos propostos t?m um bom potencial de servir como marcadores moleculares eficientes para identifica??o acurada de amostras de carn?voros ao n?vel de esp?cie. / DNA sequences for species-level identification of biological materials have achieved considerable popularity in the last few years, especially in the context of the DNA barcoding initiative. Species assignment of biological samples such as hairs, feathers, pelts and particularly faeces is a crucial step for those interested in studying ecology and evolution of many species with these samples. This is especially the case for carnivores, whose elusive habits and low densities highlight the importance of studies based on noninvasive samples. However, the current lack of standardized assays for carnivore identification often poses challenges to the large-scale application of this approach, as well as the cross-comparison of results among sites. Here we evaluate the potential of two short (<250 pb) mitochondrial DNA (mtDNA) segments located within the genes ATP synthase 6 and cytochrome oxidase I as standardized markers for carnivore identification. Between one and eleven individuals of 66 carnivore species were sequenced for one or both of these mtDNA segments and analyzed using three different approaches (tree-based, distance-based and character-based), in conjunction with sequences retrieved from public databases. In most cases, conspecific individuals had lower genetic distances from each other relative to other species, resulting in diagnosable monophyletic clusters. Notable exceptions were the more recently diverged species, some of which could still be identified using diagnostic character attributes, species-specific haplotypes, or by reducing the geographic scope of the comparison (restricting the analysis to a single zoogeographic region). Additional in silico analyses using a short cytochrome b segment frequently employed in carnivore identification were also performed aiming to compare performance to that of our two focal markers. We then tested the performance of these segments in the identification of carnivore faeces via three case studies: (i) felid faeces collected in a controlled zoo experiment, aimed at assessing whether DNA from rabbit prey would contaminate the resulting sequences; (ii) field-collected faeces from the Brazilian Cerrado presumed to be from maned wolves and containing prey remains (hairs, bones, feathers), aimed at investigating the efficiency of predator identification and occurrence of prey DNA interference; and (iii) field-collected scats from an Atlantic Forest study site, also addressing the issue of PCR success rate and identification efficiency. In spite of some relevant differences in some aspects of their performance, our results indicate that both of our focal segments have a good potential to serve as efficient molecular markers for accurate species-level identification of carnivore samples.

Phlebotomine sand flies (Diptera, Psychodidae) are small blood sucking insects which are only proven vectors of neglected tropical disease called leishmaniasis. Sand flies of the genus Phlebotomus act as vectors in the Old World and those of genus Lutzomyia are vectors in the New World. However, not all of them are capable of transmitting the disease. It is therefore crutial to conclusively determinate the species and have up-to-date knowledge of their natural occurence. Routine identification based on morphological characters is challenging due to intraspecific variability of these or their possible damage during the capture and preparation. In adition, correct assessment of distinctive cahracters, especially for females, is difficult and requires certain expertise. Thus, approaches of molecular taxonomy have been recently increasingly used for sand flies species determination. This thesis presents usage of DNA sequencing and MALDI-TOF protein profiling for routine determination of sand flies caught in southeastern Europe and adjacent regions, where we have lack of information of present sand flies species. Another part of this thesis focus on closer examination of relations between selected closely related species within species complexes. Key words: Phlebotomus, molacular identification, DNA...

Introdução: O gênero Acinetobacter sp apresenta considerável heterogeneidade possuindo inúmeras espécies. Atualmente, 23 espécies já foram nomeadas e nove outras espécies já foram descritas. Quatro destas espécies possuem contextos clínicos e epidemiológicos diferentes, no entanto são agrupadas em um complexo denominado Acinetobacter calcoaceticus-Acinetobacter baumannii (ABC) devido à sua similaridade genética e fenotípica. Diversos testes para diferenciação do complexo já foram descritos, porém a maioria não pode ser realizado na rotina laboratorial, pois são caros e laboriosos. Objetivos: Avaliar um método rápido e viável na rotina laboratorial, capaz de diferenciar as espécies do complexo ABC; Determinar a prevalência das diferentes espécies do complexo ABC; Avaliar o perfil de suscetibilidade aos antimicrobianos nas diferentes espécies. Métodos: Foram analisadas 118 amostras de dois hospitais de Porto Alegre-RS através do método Multiplex PCR para o gene gyrB e posteriormente confirmadas pelo padrão-ouro: sequenciamento do 16S-23S ITS. O perfil de suscetibilidade foi realizado através de microdiluição em caldo. Resultados: Das 118 amostras identificadas inicialmente como Acinetobacter sp., a grande maioria dos isolados (106 -89.9%) foram identificados como A. baumannii; mas doze isolados foram identificados como sendo das demais espécies do complexo ABC: 6 (5.1%) A. nosocomialis, 5 (4.2%) A. pittii, e 1 (0.8%) A. genoespécie 10, através da técnica de Multiplex PCR. Todos os resultados foram confirmados por sequenciamento. A. baumannii apresentou um elevado nível (72,6%) de resistência ao imipenem em comparação com as outras espécies seguido da espécie A. nosocomialis que apresentou metade de seus isolados resistentes. Todas as espécies apresentaram baixos índices (inferior a 7,5%) de resistência à Polimixina B e Tigeciclina. Conclusão: O Multiplex PCR para o gene gyrB apresentou resultados fidedignos quando comparados ao padrão-ouro, demonstrando, assim, ser um método confiável para a identificação das espécies do complexo ABC. Outras espécies, além de A. baumannii, ABC podem apresentar percentuais significativos de resistência ao imipenem. / Background: Introduction: The genus Acinetobacter sp presents considerable heterogeneity possessing numerous species. Currently, 23 species have been named and nine other species have been described. Four of these species have different clinical and epidemiological contexts, but are grouped in a complex called Acinetobacter calcoaceticus-Acinetobacter baumannii (ABC) due to their genetic and phenotypic similarity. Several tests for differentiation of the complex have been described, but most can not be performed routinely in the laboratory, they are expensive and laborious. Objectives: To evaluate a fast and feasible in routine laboratory able to differentiate the species of the complex ABC; determine the prevalence of different species of the complex ABC; evaluate the antimicrobial susceptibility profile of the different species. Methods: We analyzed 118 samples from two hospitals in Porto Alegre-RS by the method of Multiplex PCR for gene gyrB and subsequently confirmed by the gold standard: sequencing the 16S-23S ITS. The susceptibility profile was performed by microdilution. Results: Of the 118 samples initially identified as Acinetobacter sp. The great majority of isolates (106 -89.9%) were identified as A. baumannii, but twelve isolates were identified as being from other species of the complex ABC: 6 (5.1%) A. nosocomialis, 5 (4.2%) A. pittii, and 1 (0.8%) A. genospécie 10 by Multiplex PCR technique. All results were confirmed by sequencing. A. baumannii showed a high level (72.6%) of imipenem resistance in comparison with the other species followed by the species A. nosocomialis showed that half of his resistant isolates. All species showed low levels (less than 7.5%) of resistance to Polymyxin B and Tigecycline. Conclusion: Multiplex PCR for gene gyrB results presented totally reliable when compared to the gold standard, demonstrating thus be a safe method for the laboratory. Other species besides A. baumannii, ABC may have significant percentages of resistance to imipenem.

Scrapter Lepeletier de Saint-Fargeau & Audinet-Serville, 1828 (Hymenoptera: Aculeatea: Anthophila: Colletidae) is a genus of solitary bees largely endemic to southern Africa. This dissertation investigated the phylogenetic systematics of the genus. Eleven new species of Scrapter are described, principally from the Succulent Karoo biome of South Africa, bringing the total number of species in the genus to 42. An updated dichotomous key to facilitate identification is provided. The previously unknown females of S. albifumus Eardley and S. amplispinatus Eardley are also described. The genus is recorded from outside southern Africa for the first time with the collection of S. nitidus (Friese) in Kenya. This constitutes a significant range extension of the genus. The taxonomic status of five species described by Cockerell in 1944, and subsequently overlooked, is addressed. They are all found to be synonyms of other Scrapter species, except one, which is found to be a Ctenoplectrina species (Apidae: Apinae: Ctenoplectrini). The new synonymies are: S. subincertus Cockerell = S. niger Lepeletier de Saint-Fargeau & Audinet-Serville; S. brunneipennis Cockerell = S. niger Lepeletier de Saint-Fargeau & Audinet-Serville; S. merescens Cockerell = S. leonis Cockerell; S. sinophilus Cockerell = S. algoensis (Friese). Scrapter ugandica Cockerell becomes Ctenoplectrina ugandica (Cockerell) as a new combination. Investigation of selected morphological features (e.g. postmentum, facial fovea, galea) revealed much diversity in Scrapter. The monophyly of Scrapter is not supported by unambiguous apomorphies, but is defensible by the congruence of various qualitative characters (e.g. premental fovea, T2 fovea, hindleg and sternal scopa in [females], two submarginal cells). A cladistic analysis using 25 morphological characters recovered numerous most parsimonious trees under both equal- and successive-weighting. To aid in resolution, several taxa known from only one sex or from very limited material, and with many unknown states, were deleted from the matrix. Analysis using this reduced matrix under equal- and successive-weighting resulted in better resolution, although with low consistency index values. Several subclades were common to both cladograms, and likely represent monophyla. The low consistency indices and general lack of unique synapomorphies upholding these subclades, however, dictated against making any classificatory re-arrangements. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2006.