Made available in DSpace on 2016-12-08T16:24:15Z (GMT). No. of bitstreams: 1 PGCA13MA115.pdf: 2445554 bytes, checksum: 3ef0e43758000b9d20b09e8f978e7502 (MD5) Previous issue date: 2013-07-10 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The species of the genus Eimeria are responsible for the greatest economic impact among the parasitic diseases that affect broilers. The control of coccidiosis is mainly performed with anticoccidial, however, the development of resistance to these drugs, together with the public opinion against the use of drugs in food, they tend to promote the use of vaccines, increasing the importance of the identification of species of Eimeria circulating. Aiming to identify the species of Eimeria and study the occurrence of these species in poultry farms in the southern state of Santa Catarina, samples were collected (pool of feces) in 21 municipalities, from 251 flocks of broilers aged 28 to 48 days. The oocysts were recovered by techniques of filtration, centrifugation and flotation using supersaturated solution of NaCl and quantified using a Neubauer chamber. The samples were stratified by age of the birds in three intervals (28 to 34, 35 to 41 and 42 to 48). The species were identified by Multiplex PCR technique. Amplicons of the seven species of Eimeria originating from the PCR positive samples have been cloned and sequenced. The occurrence of the genus Eimeria was 96.02%. It was found that in chickens aged 42 to 48 days the average number of oocytes was significantly reduced when compared to the intervals age of 28 to 34, and 35 to 41 days (p<0.05). Were found Eimeria acervulina (63.3%), Eimeria maxima (63,7%), Eimeria tenella (54.6%), Eimeria praecox (25.1%), Eimeria mitis (38.6%), Eimeria necatrix (24.3%) and Eimeria brunetti (13.1%). The average number of species detected by property was 2.96 and was the most frequent combination of E. acervulina, E. maxima and E. tenella (9.16%). The sequencing of the clones confirmed the specificity and effectiveness of the Multiplex PCR technique for identification of species of Eimeria. It can be concluded that in the South of the State, prophylactic measures should consider the control of the seven Eimeria species that parasitize chickens / As espécies do gênero Eimeria são responsáveis pelo maior impacto econômico dentre as doenças parasitárias que acometem frangos de corte. O controle da coccidiose é realizado principalmente com anticoccidianos, porém, o desenvolvimento da resistência a esses fármacos, aliada à opinião pública contra o uso de drogas na ração, tendem a fomentar a utilização de vacinas, aumentando a importância da identificação das espécies de Eimeria circulantes. Com os objetivos de identificar as espécies de Eimeria e estudar a ocorrência destas em instalações avícolas da mesorregião Sul do estado de Santa Catarina, foram coletadas amostras (pool de fezes) em 21 municípios, provenientes de 251 lotes de frangos de corte com idade entre 28 a 48 dias. Os oocistos foram recuperados por meio de técnicas de filtragem, centrifugação e centrífugo-flutuação utilizando solução hipersaturada de NaCl e quantificados utilizando câmara de Neubauer. As amostras foram estratificadas, por idade das aves em três intervalos (28 a 34; 35 a 41 e 42 a 48). As espécies foram identificadas por meio da técnica de Multiplex PCR. Amplicons das sete espécies de Eimeria originados da PCR de amostras positivas foram clonados e sequenciados. A ocorrência do gênero Eimeria foi de 96,02%. Verificou-se que em frangos com idade entre 42 a 48 dias a média do número de oocistos foi significativamente menor quando comparada com os intervalos de idade de 28 a 34 e 35 a 41 dias (P<0,05). Foram encontradas Eimeria acervulina (63,3%), Eimeria maxima (63,7%), Eimeria tenella (54,6%), Eimeria praecox (25,1%), Eimeria mitis (38,6%), Eimeria necatrix (24,3%) e Eimeria brunetti (13,1%). O número médio de espécies detectadas por propriedade foi de 2,96 e a associação mais frequente foi de E. acervulina, E. maxima e E. tenella (9,16%). Por meio do sequenciamento dos clones confirmou-se a especificidade e eficácia da técnica de Multiplex PCR para a identificação das espécies de Eimeria. Concluí-se que na mesorregião Sul do Estado, a adoção de medidas profiláticas deve considerar o controle das sete espécies de Eimeria que parasitam frangos

Submitted by Franciele Moreira (francielemoreyra@gmail.com) on 2017-10-26T13:06:49Z No. of bitstreams: 2 Tese - Ricardo Goulart Rodovalho - 2017.pdf: 4249347 bytes, checksum: b731c79e9585351e61ca0e415749d4b1 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2017-10-26T14:48:15Z (GMT) No. of bitstreams: 2 Tese - Ricardo Goulart Rodovalho - 2017.pdf: 4249347 bytes, checksum: b731c79e9585351e61ca0e415749d4b1 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-10-26T14:48:15Z (GMT). No. of bitstreams: 2 Tese - Ricardo Goulart Rodovalho - 2017.pdf: 4249347 bytes, checksum: b731c79e9585351e61ca0e415749d4b1 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-09-05 / Fundação de Amparo à Pesquisa do Estado de Goiás - FAPEG / The main scope of the current study was to develop a short tandem repeat (STR) multiplex system, made up of 22 highly informative loci, for application in forensic genetics. The system comprised of 21 polymorphic autosomal short tandem repeat loci, namely D3S1358, TH01, D21S11, D18S51, Penta E, D5S818, D13S317, D7S820, D16S539, CSF1PO, Penta D, vWA, D8S1179, TPOX, FGA, D2S441, D17S1301, D19S433, D18S853, D20S482 and D14S1434, and the amelogenin gene locus. Strategies were developed to overcome the challenges involved in creating a multiplex system. Based on the literature and available databases, STR loci were selected to obtain discriminatory markers, and followed specific criteria for this purpose. Primers were designed using the Primer3 software and the AutoDimer was used to evaluate potential interactions between them. The 21 selected STR loci were validated individually and jointly, both to assess their sensitivity and to test the efficiency of the multiplex system. Statistical analyses were based on the genetic data of 450 unrelated individuals living in the State of Goiás, thus allowing the establishment of the parameters necessary to use this system. A total of 239 alleles were detected for the 21 loci in the set, allowing for a probability of identity of 4.23 x 10-25 to be obtained. The combined power of discrimination was 0.999999999999999999999999 and the combined power of exclusion was 0.99999. Upon complete validation of the entire system, this multiplex assay was considered to be a powerful tool for application in human identification by DNA analysis. / O escopo principal do atual estudo foi o desenvolvimento de um sistema multiplex de loci STR, composto por 22 loci altamente informativos para aplicação em genética forense. O sistema compreendeu 21 loci STRs polimórficos autossômicos, a seguir D3S1358, TH01, D21S11, D18S51, Penta E, D5S818, D13S317, D7S820, D16S539, CSF1PO, Penta D, vWA, D8S1179, TPOX, FGA, D2S441, D17S1301, D19S433, D18S853, D20S482 e D14S1434, e o locus do gene da amelogenina. As estratégias foram desenvolvidas para superar os desafios envolvidos na criação de um sistema multiplex. Com base na literatura e nas bases de dados disponíveis, os loci STRs foram selecionados para obter marcadores discriminatórios e seguiram critérios específicos para esse fim. Os primers foram projetados usando o software Primer3, e o AutoDimer foi usado para avaliar potenciais interações entre eles. Os 22 loci selecionados foram validados individualmente e em conjunto, tanto para avaliar sua sensibilidade quanto para testar a eficiência do sistema multiplex. As análises estatísticas foram baseadas nos dados genéticos de 450 indivíduos não relacionados e residentes no estado de Goiás, permitindo assim estabelecer os parâmetros necessários para a utilização do sistema. Um total de 239 alelos foram detectados para os 21 loci do conjunto, permitindo obter uma probabilidade de identidade de 4,23 x 10-25. O poder combinado de discriminação foi 0,999999999999999999999999 e o poder combinado de exclusão foi 0,99999. Após a validação completa de todo o sistema, este ensaio de multiplex foi considerado uma ferramenta poderosa para aplicação na identificação humana pela análise do DNA.

La technique de spectroscopie basée sur la diffusion Raman Stokes spontanée est un procédé standard employé dans de nombreux domaines allant de la thermodynamique à la médecine, en passant par la science des matériaux. À la faveur d'un échange d'énergie inélastique, elle permet de déterminer les fréquences des vibrations moléculaires présentes dans un objet. On peut ainsi remonter à l'identification des molécules et ainsi caractériser l'objet d'étude sans utiliser de marqueur spécifique. Cette méthode est néanmoins affligée de défauts. Outre la présence d'un signal de fluorescence qui peut submerger la réponse Raman, le désavantage majeur est le long temps d'exposition que requière cette technique. Dans le cas d'étude d'échantillon biologique, cela proscris son usage pour des mesures de microspectroscopie : la cartographie spectrale d'objet microscopique. Afin de pallier ce problème, de nouvelles techniques ont été développées. C'est le cas de la spectroscopie employant la diffusion Raman anti-Stokes Cohérente (ou CARS pour Coherent Anti-Stokes Raman Scattering). Du fait de sa cohérence et de sa directivité le signal anti-Stokes affiche une intensité 10^5 to 10^6 fois plus importante que dans le cas de la diffusion Raman spontanée, ce qui permet alors d'abaisser le temps d'exposition à un niveau tolérable pour les objets biologiques lors d'une mesure de microspectroscopie. De plus, le caractère anti-Stokes du signal l'épargne de la contribution de la fluorescence. Pourtant, un défaut majeur limite encore l'utilisation de cette technique : le bruit de fond non résonant. Ce phénomène peut diminuer, voir noyer la contribution résonante qui porte l'information. Cette thèse a permis le développement de techniques CARS autorisant une réduction du bruit de fond non résonant. Pour ce faire un dispositif de spectroscopie CARS multiplex (M-CARS) en configuration copropagative a été construit. Ses capacités sont illustrées par des mesures spectrales d'échantillons minéral, végétal et biologique. À partir de ce système, il a été établi une méthode innovante permettant de discriminer le signal résonant du bruit non résonant en utilisant un champ électrique continu. Il est aussi démontré la mise en place d'un procédé qui a permis de mener la première mesure de microspectroscopie M-CARS en configuration contrapropagative sur un échantillon biologique. Cette configuration limite la collecte du signal à l'objet d'étude, empêchant ainsi l'acquisition du signal résonant et non résonant issu du solvant, principal responsable du bruit de fond non résonant lors d'une mesure CARS en configuration copropagative. / The spectroscopy technique based on spontanée Raman Stokes scattering is a standard process used in many fields spanning from thermodynamic and medicine, to materials sciences. An inelastic energy exchange permits to determinate the frequency of the molecular vibrations in an object. One can identify the molecules and thus, can characterize the object of study in a label-free way. Nevertheless, this method is afflicted with faults. Beside the presence of fluorecence that can drown the Raman answer, the main drawback is the long exposition time required. In the case of biological sample, this can prohibit the use of spontaneous Raman scattering for microspectroscopy measures: the spectral mapping of microscopic objects. To avoid this problem, new techniques have been developed. It is the case of Coherent anti-Stokes Raman scattering (CARS) spectroscopy. Due to its coherence and its directivity, the anti-Stokes signal has an intensity 105 to 106 times greater than the spontaneous Raman scattering one. The exposition time is then reduced to a tolerable level for biological objects during microspectroscopy measures. Moreover, the anti-Stokes characteristic of the signal prevents the fluorescence contribution. However, a major fault still limits the use of this technique: the nonresonant background. This phenomenon can diminish, even overwhelm the resonant contribution carrying the information. This thesis permitted the development of CARS approaches that allow the reduction of the nonresonant background. To do so, a multiplex CARS (M-CARS) spectroscopy apparatus in a forward configuration has been built. Its abilities are illustrated with spectral measures of mineral, vegetal and biological samples. Based on this system, it has been established an innovative method that can discriminate the resonant signal from the nonresonant one thanks to a static electric field. It has been also been demonstrated the development of a process that has allowed the first M-CARS microspectroscopy measure of a biological sample in a contrapropagative configuration. This setup limits the collect of the signal to the object of study, avoiding the acquisition of the resonant and resonant signals coming from the solvent, responsible for the major part of non resonant background during a CARS measure in a forward configuration.

O presente trabalho foi conduzido na escarpa sul da Serra da Mantiqueira, com o objetivo de reverter os danos ambientais causados pelo rompimento de uma adutora de água localizada a 1.700 m de altitude. Esse acidente resultou na formação de uma cicatriz na floresta da encosta, com supressão do solo e da vegetação. Para reverter os processos erosivos formados no local foi desenvolvida uma nova técnica utilizando quatro barreiras formadas por mudas de Bambusa mutiplex (Lour.), com o intuito de desviar as águas e favorecer a regeneração da vegetação. Para contenção de duas ravinas, denominadas de menor e maior, colmos de bambu (Bambusa sp) foram plantados em consórcio com B. multiplex. O projeto foi monitorado por 18 meses e sua avaliação se fez por meio de observações de campo e preenchimento de ficha técnica. Posteriormente, os dados foram aplicados a uma adaptação da Matriz de Leopold composta por cinco aspectos monitorados: erosão, regeneração da vegetação, sucesso do plantio de bambu, conservação das estruturas e funcionalidade, nos quais se procurou avaliar a eficiência das intervenções com o bambu. O resultado da Matriz possibilitou classificar qualitativa e quantitativamente as intervenções, obtendo-se cinco níveis de classificação. As Barreiras I, II e IV foram classificadas como sendo de muita alta eficiência e, a Barreira III, como de alta eficiência. A contenção da ravina maior foi classificada como média eficiência e, da menor, como de muito alta eficiência. / This work was carried out in southern escarpment of the Serra Mantiqueira, city of Pindamonhangaba within an area adjacent to the Plateau of Campos do Jordão, in São Paulo state, Brazil, and claimed as a result to reverse environmental damage caused by the rupture of an adductor of water located at 1700 m altitude. This accident resulted in the formation of a scar in the hillside forest, with its removal of soil and vegetation. To reverse the erosion processes created at the site a new technique was developed using four barriers formed by seedlings of Bambusa mutiplex (Lour.) in order to divert the waters and promote the regeneration of vegetation. For containment of two ravines, called lower and higher, stalks of bamboo (Bambusa sp.) were installed on the walls of the ravines with intercropped planting of seedlings B. multiplex. The project was monitored for 18 months and its evaluation was done through observations of field and fill in sheet. Subsequently, the data were applied to an adaptation of the Leopold Matrix composed of five monitored aspects: erosion, regeneration of vegetation, the success of bamboo plantation, preservation of the structures and functionality, which sought to evaluate the effectiveness of interventions with the bamboo. The result of the classification matrix enabled the qualitative and quantitative interventions, resulting in five levels of classification. The barriers I, II and IV were classified as being of very high efficiency, and the Barrier III as a high efficiency. The containment of the higher ravine was classified as medium average efficiency, the lower, as a very high efficiency.

Beta lactam resistance producing isolates of Enterobacteriacea and non-Enterobacteriacea have emerged since more than seventy years ago (Abraham and Chain, 1940). They are known to cause both community and hospital-acquired infections. Resistance against carbapenem is primarily mediated by the production of enzymes that destroy the beta lactam antimicrobials, which are produced by these isolates involving the expression of serine and metalobetalactamase genes KPC, IMP, VIM, NDM-1 and OXA-48. Quinolone resistance is predominanty mediated by mutations in the qnrA, qnrB, qnrS, and aac-6-Ib genes. Carbapenemase-producing organisms especially Klebsiella pneumoniae carbapenemases (KPCs) emerged as important pathogens especially among critically ill patients causing significant morbidity and mortality. This study aims to determine the prevalence and types of 15 quinolone resistance and carbapenemases genes among different isolates from patients admitted to the University Hospital of Leipzig over a period of ten months. During the period from January 2011 through October 2011, a total of 50 carbapenemases isolates were recovered from patients of the University Hospital of Leipzig/ Germany. The isolates were identified by biochemical tests and their susceptibility to antimicrobials was determined by the microbroth dilution method according to ISO standard. The KPC, IMP, VIM, OXA-48, NDM-1, and aac-6-Ib genes as well as qnrA, qnrB, and qnrS genes were detected by multiplex PCR, respectively. Results showed that KPC gene was detected in 82% of the isolates while 8% were KPC negative. The qnrA, qnrS, IMP, NDM-1, and OXA-48 genes were not detected in any of the isolates while qnrB and VIM genes were found in 2%. On the other hand, aac-6-Ib gene was the most prevalent gene among the study isolates and composed a percentage of 96%. Results also showed that KPC, and aac-6-Ib genes were detected in isolates collected from urine, blood, wounds, swabs, sputum, tracheal secretions, biopsies, and anal smears, while VIM gene was detected in one isolate collected from blood. The qnrB gene was found in one isolate collected from urine specimen. The wide spread of carbapenem and quinolone resistance-producing organisms is a critical problem that complicates the treatment of infections resulting from these organisms. Necessary measures must, therefore, be taken to limit their spread, which include appropriate antibiotic treatment, control of hospital infections, observe of personal hygiene, and the use of appropriate methods of sterilization and disinfection to prevent the dissemination of these organisms. Keywords: Resistance, carbapenemases, QRDR, multiplex PCR, antimicrobials

No description available.

No description available.

Made available in DSpace on 2014-07-29T15:16:37Z (GMT). No. of bitstreams: 1 AnaClaraferraz.pdf: 1870286 bytes, checksum: d17e1fe96ee32ca1852c51743beff160 (MD5) Previous issue date: 2010-01-12 / The progress of Molecular Biology and Genetics provided the appearance of several molecular markers that detect the genetic polymorphism directly at DNA. Among these markers are the microsatellites (SSR), which are distinguished by their high degree of polymorphism. The use of these markers for individual genotyping has evolved into multiplex systems, which allow many SSR fragments to be detected and analyzed simultaneously. Currently there are several articles in literature discussing the criteria to be used in the primer design for use in PCR, as well as various softwares are available for this end. However, there are few studies and tools for the analysis of compatibility between pairs of primers for use in multiplex systems, where multiple fragments are simultaneously amplified using PCR. This paper evaluated different criteria for compatibility between pairs of primers. A set of 74 combinations of pairs of primers, involving the amplification of 94 SSR loci were evaluated in duplex systems. The same combinations were evaluated according to different criteria, including the degree of complementarity between primers, the magnitude of differences of denaturation temperatures (Tm) and the tendency to annealing between pairs of primers based on the Gibbs free energy resulting from the association between them. The comparison between the different criteria allowed the identification of a set of criteria with positive predictive value equal to 94%. These criteria were implemented for use in a software called Multiplexer, which from the analysis in sequence of pairs of primers, suggests compatible combinations for use in multiplex genotyping systems. Using this tool can significantly reduce the costs related to laboratory activities for genotyping using PCR. / Os avanços da Biologia Molecular e da Genética proporcionaram o surgimento de diversos marcadores moleculares que detectam o polimorfismo genético diretamente no DNA. Entre estes marcadores se encontram os microssatélites (SSR), que se destacam pelo seu elevado grau de polimorfismo. O uso desses marcadores para fins de genotipagem individual tem evoluído para sistemas multiplex, os quais permitem que vários fragmentos SSR sejam detectados e analisados simultaneamente. Atualmente são abundantes na literatura artigos que discutem os critérios a serem utilizados no desenho de pares de primers para aplicação em PCR, bem como estão disponíveis diversos softwares para este fim. No entanto, ainda são escassos os estudos e ferramentas destinados à análise de compatibilidade entre pares de primers para aplicação em sistemas multiplex, onde vários fragmentos são amplificados simultaneamente por PCR. Neste trabalho são avaliados diferentes critérios de compatibilidade entre pares de primers. Um conjunto de 74 combinações de pares de primers, envolvendo a amplificação de 94 locos SSR foram avaliados em sistemas duplex. As mesmas combinações foram avaliadas segundo diferentes critérios, incluindo o grau de complementariedade entre primers, magnitude das diferenças de temperaturas de desnaturação (Tm) e a tendência ao anelamento entre pares de primers com base na energia livre de Gibbs resultante da associação entre eles. A comparação entre os diferentes critérios permitiu a identificação de um conjunto de critérios com valor preditivo positivo igual a 94%. Estes critérios foram implementados para utilização em um software denominado Multiplexer, que a partir da análise de sequências de pares de primers, sugere combinações compatíveis para a utilização em sistemas de genotipagem multiplex. O uso dessa ferramenta pode reduzir consideravelmente os custos laboratoriais relativos às atividades de genotipagem utilizando PCR.

Cette thèse présente une nouvelle méthodologie pour analyser des réseaux. Nous développons l'intrication d'un réseau multiplex, qui se matérialise sous forme d'une mesure d'intensité et d'homogénéité, et d'une abstraction, le réseau d'interaction des catalyseurs, auxquels sont associés des indices d'intrication. Nous présentons ensuite la mise en place d'outils spécifiques pour l'analyse visuelle des réseaux complexes qui tirent profit de cette méthodologie. Ces outils présente une vue double de deux réseaux,qui inclue une un algorithme de dessin, une interaction associant brossage d'une sélection et de multiples liens pré-attentifs. Nous terminons ce document par la présentation détaillée d'applications dans de multiples domaines. / When it comes to comprehension of complex phenomena, humans need to understand what interactions lie within them.These interactions are often captured with complex networks. However, the interaction pluralism is often shallowed by traditional network models. We propose a new way to look at these phenomena through the lens of multiplex networks, in which catalysts are drivers of the interaction through substrates. To study the entanglement of a multiplex network is to study how edges intertwine, in other words, how catalysts interact. Our entanglement analysis results in a full set of new objects which completes traditional network approaches: the entanglement homogeneity and intensity of the multiplex network, and the catalyst interaction network, with for each catalyst, an entanglement index. These objects are very suitable for embedment in a visual analytics framework, to enable comprehension of a complex structure. We thus propose of visual setting with coordinated multiple views. We take advantage of mental mapping and visual linking to present simultaneous information of a multiplex network at three different levels of abstraction. We complete brushing and linking with a leapfrog interaction that mimics the back-and-forth process involved in users' comprehension. The method is validated and enriched through multiple applications including assessing group cohesion in document collections, and identification of particular associations in social networks.

As doenças respiratórias são responsáveis, na suinocultura, por grandes perdas econômicas e entre os agentes destacam-se os micoplasmas. Foram coletadas 126 amostras de muco nasal/tonsilar, e fragmentos de 78 pulmões, 2 de traquéia e 2 de tonsila. No isolamento foi utilizado o meio FRIIS modificado, na identificação, a Multiplex-PCR e na tipificação, a PFGE e sequenciamento do gene 16S rRNA. Foram obtidos 59 isolados identificados como M. hyopneumoniae (1,70%), M. flocculare (3,40%) e M. hyorhinis (94,90%). A PFGE dos isolados de M. hyorhinis, resultou em 10 e 9 perfis com a enzima AvaI e XhoI, respectivamente. O sequenciamento do gene 16S rRNA dos isolados M. hyorhinis apresentaram baixo polimorfismo quando comparados entre si e com a cepa de referência. A sequência do gene 16S rRNA do isolado de M. hyopneumoniae, quando comparada a seqüência da cepa J e os isolados 7448 e 232, resultaram em polimorfismo. O M. hyopneumoniae continua sendo o mais difícil de isolar. Os dendrogramas obtidos da PFGE resultaram em grande heterogeneidade entre os isolados de M. hyorhinis. O seqüenciamento do gene 16S rRNA permitiu o estudo de variabilidade interespecífica e intraespecífica dos isolados de micoplasmas. / Economic losses in swine production are common due the respiratory diseases in these animals. M. hyopneumoniae and M. hyorhinis are the most frequent microbial agents. The aim of this study was recover this isolates in FRIIS medium, indentify them by Multiplex PCR and detect their genotypic variations by PFGE and sequencing the 16s rRNA gene. One hundred twenty six swabs from tonsil and nasal mucus of swine with respiratory disturbances were analyzed. It was included 78 lungs, two trachea and two tonsils. It was obtained 59 isolates; 1.70% of M. hyopneumoniae, 3.40% of M. flocculare and 94.90% of M. hyorhinis. The PFGE of M. hyorhinis, allowed obtain 10 profiles with enzyme AvaI and nine profiles with XhoI. A low polymorphism of gene 16sRNS was detected in M. hyorhinis isolates when compared with the type strain at the GenBank. The M. hyopneumoniae isolates resulted in polymorphisms when comparated with strain J, 7448 and 232. M. hyopneumoniae is still the most difficult to isolate. M. hyorhinis isolates of different herds showed a large heterogenicity with enzymes AvaI e XhoI. The sequencing of gene 16S rRNA allowed analyse the interespecífic and intraespecífic variations of mycoplasmas isolated.