Made available in DSpace on 2016-06-02T20:20:34Z (GMT). No. of bitstreams: 1 TeseSMC.pdf: 1475573 bytes, checksum: 4f80d07f9aea9ed3c478641a4ab37393 (MD5) Previous issue date: 2004-06-29 / The PIT1 gene codes for the pituitary transcription factor Pit-1, which is critical for the activation of the expression of prolactin and growth hormone genes, in addition to participating in the activation of PIT1 itself. The objective of the present study was to investigate the effect of HinfI polymorphism of this gene on meat production traits. The sample consisted of 509 Canchim animals born between 1998 and 2000. The sample included a traditional line (GG1) consisting of 5/8 Charolais genome breed and 3/8 of the Zebu breeds, Nelore, Guzerá and Indubrasil, and a new line (GG2) with 21/32 Charolais genome and 11/32 of the Nelore breed. The genotypes for the PIT1 gene were identified by PCR-RFLP. Genotype effect on phenotypic values for birth weight (BW), standardized weaning weight (W240) and at 12 month of age weight (W365), and on average daily weight gain to the born at weaning (GMND) and to the weaning at 12 month of age (GMD12), were analyzed by the least squares method. The results revealed significant effects of the interaction between genetic animal group and PIT1 genotype on W240, GMND and GMD12 (P < 0.05). The differences in mean least squares between genotype ( / ) and genotypes (+/+) and (+/ ) for W240 and GMND were significant in GG2 (P < 0,01 and p < 0,05, respectively), revealing superiority of the ( / ) genotype on that characters. The means for genotypes (+/+) and (+/ ), respectively at the W240 and GMND, did not differ from one another, suggesting a dominance effect of the HinfI (+) allele. The means effects of the HinfI ( ) allele indicated a positive effect of this allele on W240 and GMND, and the deviation estimates attributed to dominance showed a favorable effect of the HinfI ( ) allele on W240 and GMND of GG2 animals, when present in homozygosis in the genotypes of the individuals. The means of the genotypes in GG1 did not differ from one another. The difference in PIT1 behavior observed in the two genetic groups, however, may suggest the action of a quantitative trait locus linked to PIT1 segregating only in GG2 of the population studied, and not a direct effect of PIT1. / O gene PIT1 codifica o fator de transcrição pituitário Pit-1, crítico para ativar a expressão dos genes da prolactina e do hormônio de crescimento, além de participar na ativação do próprio PIT1. Esta pesquisa teve por objetivo investigar o efeito do polimorfismo HinfI desse gene sobre caracteres de produção de carne em um rebanho da raça Canchim. A amostra foi constituída por 509 animais nascidos entre os anos de 1998 e 2000. A amostra incluiu uma linhagem tradicional (GG1) com média de 5/8 de genoma da raça Charolês e 3/8 das raças zebuínas Nelore, Guzerá e Indubrasil, e uma linhagem nova (GG2) com média de 21/32 de genoma de Charolês e 11/32 da raça Nelore. Os genótipos para o gene PIT1 foram identificados mediante técnica PCR-RFLP empregando a enzima HinfI. Os efeitos dos genótipos sobre valores fenotípicos para pesos ao nascimento (PN) e pesos padronizados à desmama (P240) e ao ano (P365), e sobre os valores de ganhos de peso médios diários do nascimento à desmama (GMND) e da desmama aos 12 meses de idade (GMD12) foram analisados pelo método dos quadrados mínimos. Os resultados revelaram efeitos significativos da interação entre grupo genético do animal e genótipo de PIT1 sobre P240, GMND e GMD12 (P < 0,05). As diferenças nas médias dos quadrados mínimos entre o genótipo ( / ) e os genótipos (+/+) e (+/ ) para P240 e para GMND foram significativas em GG2 (P < 0,01 e p < 0,05, respectivamente), revelando superioridade do genótipo ( / ). As médias dos genótipos (+/+) e (+/ ), referentes a P240 e GMND, não diferiram entre si, indicando efeito de dominância do alelo HinfI (+). As médias dos efeitos do alelo HinfI ( ) apontaram para efeito positivo deste alelo sobre P240 e GMND, e as estimativas de desvios atribuídos à dominância mostraram efeito favorável do alelo HinfI ( ), sobre P240 e GMND dos animais do grupo GG2, quando presente em homozigose nos genótipos dos indivíduos. Não houve diferenças significativas entre as médias dos três genótipos no grupo genético GG1. A diferença de comportamento de PIT1 observada nos dois grupos genéticos, entretanto, pode sugerir a ação de um QTL ( Quantitative Trait Locus) ligado ao gene PIT1 segregando apenas em GG2 da população estudada e não efeito direto de PIT1.

When paper is manufactured, deposits often form in the machines. These deposits are slimelike and can interfere with the papermaking process. The slimelike deposits are aggregates of micro-organisms, also known as biofilm. One single type of micro-organism can form a biofilm, but most biofilms consists of a mixture of several different kinds of micro-organisms and can form on about any conceivable surface. To control the aggregates of micro-organisms a slimecide is added, a so-called biocide. To examine what kind of bacteria that is included in the biofilm and also which bacteria that is killed or not killed by the biocide, Terminal Restriction Fragment Length Polymorphism analysis (T-RFLP) can be used.   In this report we examine biocides impact on biofilm produced in the laboratory.The biocides were first tested for possible interference with the PCR-step of the T-RFLP analysis. None of the tested ten biocides inhibited the PCR process the biofilm was formed on metal plates when these were lowered in a beaker with white water. Three different beakers were set up, one with addition of a biocide with active component 4,5-DICHLORO-1,2-DITHIOLONE from the start, one with the addition of the same biocide after three days and one with no addition at all of biocide. Samples were taken from the beakers and analyzed with T-RFLP.   In this report, we show that biocides affect planktonic and biofilm micro-organisms differently. There are however some micro-organisms in the biofilm that does not get affected by the biocide.   The experimental in this report is a good way of investigate the influence that biocides have on planktonic and biofilm micro-organisms, but to get even greater result the experiment should be done over a longer period of time and repeatedly.

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

Submitted by Marlene Santos (marlene.bc.ufg@gmail.com) on 2016-09-01T17:47:29Z No. of bitstreams: 2 Dissertação - Vanderlei Alves Cardoso - 2016.pdf: 2005513 bytes, checksum: f02b1c68047a6e30b76b4cd7e996de8d (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-09-05T13:05:25Z (GMT) No. of bitstreams: 2 Dissertação - Vanderlei Alves Cardoso - 2016.pdf: 2005513 bytes, checksum: f02b1c68047a6e30b76b4cd7e996de8d (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2016-09-05T13:05:25Z (GMT). No. of bitstreams: 2 Dissertação - Vanderlei Alves Cardoso - 2016.pdf: 2005513 bytes, checksum: f02b1c68047a6e30b76b4cd7e996de8d (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2016-07-15 / There are several factors that may affect the milk yield in cattle, among which the environmental characteristics and the genetic profile are the most important. The use of tools for genetic evaluation of animals, in particular the identification of SNPs, which are able to interfere with the production capacity, is being widely studied and used in animal production. In this sense, the objective of this study was to investigate the distribution of polymorphic variants of the promoter region and éxon8 in gene of growth hormone receptor (GHR) in Holstein cows in milk. They used data from lactations and milk composition in the first and second lactation of 106 cows in the municipality of Cristalina, Goiás. The blood samples were collected and the genomic material was purified (DNA). The genotypes were analyzed by PCR-FRLP technique using the AluI enzyme in the promoter region and Sspl to the exon8 of the GHR gene, respectively. For the data analysis of lactation and milk composition was performed analyses of varyance and Tukey test. Allelic frequencies of 47.64% were observed for AluI (-) and 52.36% for AluI (+) for the polymorphism of the promoter region, 49.53% for Ssp (-) and 50.47% for Ssp (+ ) for the polymorphism of exon8 of the gene. As to the genotypic frequencies, the promoter region had 12.26%, 70.76% and 16.98% for AluI genotypes (- / -), AluI (+/-) and AluI (+ / +) respectively. While the region of exon8 presented frequencies of 7.55%, 83.96% and 8.49% for genotypes Ssp (- / -), SspI (+/-), and SspI (+ / +) respectively. No statistically significant differences were found (P> 0.05) for the production and composition of milk on the effects of polymorphisms of the promoter region (AluI) and exon 8 (SspI) of the GHR gene. The composition EST and ESD were higher for SspI genotypes (- / -), but with significance level P = 0.06 and P = 0.05 respectively for EST and ESD. The interaction between the two polymorphisms and their effects on milk production and composition, no statistically significant differences were observed (P> 0.05). / Existem vários fatores que podem interferir na produção de leite em bovinos, entre os quais, as características ambientais e o perfil genético são os mais importantes. O uso de ferramentas para avaliação genética dos animais em especial a identificação de SNPs, os quais são capazes de interferir na capacidade produtiva, está sendo amplamente estudado e utilizado na produção animal. Neste sentido, o objetivo do presente trabalho foi verificar a distribuição das variantes polimórficas da região promotora e do éxon 8 no gene do Receptor do Hormônio do Crescimento (GHR) em vacas da raça Holandesa em lactação. Foram utilizados dados das lactações e composição do leite na primeira e segunda lactação de 106 vacas do município de Cristalina, Goiás. As amostras de sangue periférico foram coletadas e o material genômico foi purificado (DNA). Os genótipos foram analisados pela técnica de PCR-FRLP com o uso da enzima AluI na região promotora e SspI para o éxon 8 do gene do GHR, respectivamente. Para a análise dos dados das lactações e composição do leite foi realizada análise de variância e teste de tukey. Foram observadas frequências alélicas de 47,64% para AluI(-) e 52,36% para AluI(+), para o polimorfismo da região promotora, 49,53% para SspI(-) e 50,47% para SspI(+) para o polimorfismo do éxon 8 do gene. Quanto as frequências genotípicas, a região promotora apresentou 12,26%, 70,76% e 16,98% para os genótipos AluI(-/-), AluI(+/-) e AluI(+/+) respectivamente. Enquanto a região do éxon 8 apresentou frequências de 7,55%, 83,96% e 8,49% para os genótipos SspI(-/-),SspI(+/-) e SspI(+/+) respectivamente. Não foram encontradas diferenças estatisticamente significativas (P>0,05) para produção e composição do leite sobre o efeitos dos polimorfismos da região promotora (AluI) e éxon 8 (SspI) do gene do GHR. A composição em EST e ESD se mostraram maiores para os genótipos SspI(-/-) porem com níveis de significância P=0,06 e P=0,05 respectivamente para EST e ESD. Quanto a interação entre os dois polimorfismos e seus efeitos sobre a produção e composição do leite, não foram observadas diferenças estatisticamente significativa (P>0,05).

O melão-de-São-Caetano (Momordica charantia L) é uma cucurbitácea que ocorre em todas as regiões geográficas brasileiras. A espécie apresenta duas \"variedades\" distintas, comumente conhecidas por \"variedade silvestre\" e \"variedade cultivada\", esta última caracterizada por plantas de maior tamanho. Plantas da \"variedade selvagem\" são hospedeiras de um fitoplasma do grupo 16SrIII-J, que causa superbrotamento de ramos, nanismo generalizado, clorose, redução no tamanho de folhas e frutos. No entanto, não há informação da \"variedade cultivada\" ser hospedeira de fitoplasmas. Assim, no presente estudo, plantas sintomáticas desta \"variedade\" foram analisadas quanto à presença de fitoplasmas em seus tecidos, visando associar este tipo de patógeno à doença. Neste mesmo trabalho, os isolados do fitoplasma encontrados nas plantas sintomáticas da \"variedade cultivada\" foram molecularmente analisados em relação ao gene secY, atualmente usado como um marcador que permite identificar variações genéticas sutis. Buscou-se, com isto, identificar possíveis variantes entre os isolados. Amplificações de fragmentos de DNA a partir dos genes 16S rRNA (1,2Kb) e secY (1,6Kb) demonstraram a associação de fitoplasma do grupo 16SrIII com as plantas sintomáticas de melão-de-São-Caetano da \"variedade cultivada\". Nestas plantas, corpúsculos pleomórficos, típicos de fitoplasmas, foram visualizados no floema dos tecidos doentes, confirmando os resultados dos testes moleculares. Análises de RFLP virtual, conduzidas com o fragmento genômico correspondente ao gene secY e diversas enzimas de restrição, permitiram identificar o referido fitoplasma como pertencente ao subgrupo 16SrIIIJ. A análise filogenética evidenciou que o fitoplasma padrão do subgrupo 16SrIII-J emergiu do mesmo ramo que o fitoplasma identificado no melão-de-São-Caetano, confirmando que ambos estão estritamente relacionados. O emprego do gene secY como marcador para diferenciação fina entre fitoplasmas não apontou variabilidade genética entre os isolados do fitoplasma detectado tanto nas plantas da \"variedade cultivada\" como naquelas pertencentes à \"variedade selvagem\". Como conclusão, a \"variedade cultivada\" de M. charantia é hospedeira do mesmo fitoplasma encontrado na \"variedade selvagem\", o qual é afiliado ao subgrupo 16SrIII-J; por sua vez, o gene seY não distinguiu geneticamente os isolados do fitoplasma pertencente ao subgrupo 16SrIII-J, presentes nas plantas de melão-de-São- Caetano. / Momordica charantia is a species of the family Cucurbitaceae that occurs in all Brazilian geographic regions. This species presents two distinct \"varieties\", commonly called \"wild variety\" and \"cultivated variety\", whose plants are bigger than those belonging to the \"wild variety\". Plants of the \"wild variety\" are hosts of a phytoplasma of the group 16SrIII-J, which is associated with shoot proliferation, stunting, chlorosis, and small leaves and fruits. However, there is no information about the occurrence of phytoplasmas in plants of the \"cultivated variety\". Thus, in the present study, symptomatic plants belonging to this \"variety\" were analysed in order to associate phytoplasmas with the disease. In addition, strains of the phytoplasma found in symptomatic plants of the \"cultivated variety\" were molecularly analyzed in relation to the gene secY, which is currently used as a marker for identification fine of genetic diversity, aiming provide elements for construction of new schemes for classification of phytoplasmas. The objective was to identify distinct strains among the strains present in plants of the wild and cultivated varieties. Amplifications of DNA fragments from the genes 16Sr rRNA (1.2Kb) and secY (1.6Kb) demonstrated the association of a phytoplasma of group 16SrIII with the symptomatic plants belonging to \"cultivated variety\". These plants revealed the presence of pleomorphic bodies, typical of phytoplasmas, which was visualized in the phloem vessel from diseased tissues, confirming the results obtained by molecular assays. Virtual RFLP analysis, performed with the genomic fragment corresponding to the gene secY and various restriction enzymes, allowed to identify this phytoplasma as a member of the subgroup 16SrIII-J. Phylogenetic analysis revealed that the phytoplasma used as reference for 16SrIII-J subgroup and the phytoplasma identified in plants of M. charantia emerged from the same branch, confirming that both phytoplasmas are closely related. The gene secY, used as marker for fine differentiation of phythoplasmas, did not show genetic variability among the strains of the phytoplasma detected both plants belonging to \"cultivated variety\" and \"wild variety\". In conclusion, plants of the \"cultivated variety\" are hosts of the same phytoplasma found in the \"wild variety, which is affiliated with the 16SrIII-J subgroup; in addition, the gene secY did not distinguish isolates of the phytoplasma belonging to 16SrIII-J subgroup, present in plants of M. charantia.

Estudou-se Microsporum gypseum de diferentes fontes regiões geográficas do Brasil.Os fungos foram isolados do solo, de animais e obtidos em coleções de cultura. Em 692 amostras de solo e a taxa de recuperação foi de 19,2%. A atividade queratinolítica e elastinolítica foi avaliada quantitativamente em 138 amostras de M. gypseum. A biotipagem molecular foi realizado através da técnica de PCR-RFLP com a enzima MvaI. O seqüenciamento da região ITS1 do rDNA foi realizado em amostras representativas. M. gypseum de amostras clínicas e do solo mostraram diferenças quantitativas significantes na expressão das enzimas. A PCR-RFLP para a região ITS1 não mostrou diferença. Pelo seqüenciamento foram obtidas duas espécies sexuadas (A. gypseum e A. incurvatum). As atividades enzimáticas sugerem um importante papel na patogenicidade e uma provável adaptação desta espécie de fungo ao parasitismo animal. A análise dos resultados pode auxiliar a identificação e o conhecimento de mecanismos de virulência destes fungos. / Microsporum gypseum from different geographical sources and regions of Brazil were studied. The fungi were isolated from the soil, animals and obtained in collections of culture. In 692 samples of soil, the recovery rate was 19.2%. The keratinolytic and elastinolytic activities were quantitatively performed on 138 samples of M. gypseum. The molecular biotyping was carried out by the technique of PCR-RFLP with the enzyme MvaI. The sequencing of the region ITS1 rDNA was carried out on representative samples. Samples of M. gypseum from clinical isolates and from soil showed significant quantitative differences in the expression of enzymes. The PCR-RFLP for the ITS1 region showed no difference. For the sequencing was obtained two sexual species (A. gypseum and A. incurvatum). The enzyme activities suggest an important role in pathogenicity and a likely adjustment of this kind of parasitic fungus to feed. The results may help the identification and knowledge of mechanisms of virulence of these fungi.

>Magister Scientiae - MSc / The extreme conditions of Antarctic desert soils render this environment selective towards a diverse range of psychrotrophic microbial communities. Cracks and fissures in translucent quartz rocks permit an adequate amount of penetrating light, sufficient water and nutrients to support cryptic microbial development. Hypolithons colonizing the ventral surface of these quartz rocks have been classified into three types: cyanobacterial dominated (Type I),moss dominated (Type II) and lichenized (Type III) communities. Eukaryotic microbial communities were reported to represent only a minor fraction of Antarctic communities. In this study, culture independent techniques (DGGE, T-RFLP and clone library construction) were employed to determine the profile of the dominant eukaryotes, fungi and microalgae present in the three different hypolithic communities. The 18S rRNA gene (Euk for eukaryotes), internal transcribed spacer (ITS for fungi) and microalgal specific regions of the 18S rRNA gene, were the phylogenetic markers targeted for PCR amplification from hypolith metagenomic DNA. Results suggest that the three hypolith types are characterized by different eukaryotic, fungal and microalgal communities, as implied by nMDS analysis of the DGGE and T-RFLP profiles. Sequence analysis indicates close affiliation to members of Amoebozoa, Alveolata, Rhizaria (general eukaryote), Ascomycota (fungal) and Streptophyta (microalgal). Many of these clones may represent novel species. This study demonstrates that Dry Valley hypolithons harbour higher eukaryote diversity than previously recognised.Each hypolithon is colonized by specialized microbial communities with possible keystone species. The ecological role of the detected microorganisms in the hypolith environment is also theorized, and a trophic hierarchy postulated.

[EN] Acetic acid bacteria (AAB) belong to the Acetobacteriaceae family, within the ¿-Proteobacteria class. These Gram-negative elliptical or cylindrical microorganisms occur in isolation, in pairs or in chains. They have polar flagellar or peritrichous motility. They are non-spore forming, cathalase positive and oxidase negative. They have an obligate aerobic metabolism, with oxygen as the terminal electron acceptor. Currently, the Acetobactereaceae family includes 19 genera and 72 species. Sugar and alcohol oxidising ability result in an accumulation of organic acids as final products, which is widely used in the food industry to produce vinegar from wine and fruits. The objective of this thesis was to isolate and identify AAB in blueberry and citrus epiphyt flora from plants grown in the Salto Grande region (Entre Ríos, Argentina) in order to detect the most suitable bacteria for biotechnological processes such as vinegar. 36 AAB were isolated from these fruits using enrichment techniques and plate isolation. Enrichment broths containing ethanol and acetic acid enabled the maximum recovery of AAB due to the fact that these components promote their growth. Fermented juice yielded a larger number of AAB compared to fresh fruit or peel, due to ethanol occurrence as it is an alcohol-resistant microorganisms selecting agent. Biochemical tests allowed bacteria differentiation at genus level. Six were identified: 13 bacteria isolates were identified as Acetobacter, 5 as Gluconobacter, 7 as Asaia, 5 as Acidomonas, 1 as Gluconacetobacter and 5 as Saccharibacter. Subsequently, those bacteria (Acetobacter, Gluconobacter and Gluconacetobacter genera) that could be used for the development of a suitable starter culture for the bio-oxidation of musts alcoholics obtained from these fruits, in order to obtain vinegars, were identified to specie level. Molecular techniques, such as RFLP-PCR of the 16S rDNA and RFLP-PCR of the 16S-23S rDNA intergenic spacer, were utilised. The former enabled the identification of the 16 AAB under study. C7, C8, A80, A160 and A180 isolates were identified as G. frateurii, C1, C2, C3, C4, C5, C6, A70 and A210 isolates as A. pasteurianus, A50 and A140 isolates as A. tropicalis and C9 isolate as A. syzygii. 16S-23S PCR-RFLP technique validated identifications performed using 16S; however, C1 showed a different restriction pattern from the first identification. Genus 16S partial sequencing solved this discrepancy. Growth dynamics and acetic acid production capacity were studied in the isolates in order to determine the most suitable ones to acetify fruits musts. Growth was assessed in different ethanol and acetic acid concentrations as high cell count values (109 cells/mL) are essential for the inoculum to start vinegar production, in order to achieve a short lag phase and therefore reduce process times. Three ethanol concentrations were assayed (4%, 6% and 8%) to study acetic acid production capacity. Results showed that cultures identified as A. pasteurianus (A210 and C1), A. syzygii (C9) and G. frateurii (A80) could be used as inoculum to produce vinegars, due to their high acetic acid production capacity when ethanol concentration values are 4% and 6% v/v. Finally, the most suitable culture preservation method was determined to maintain purity and activity through time. AAB may be lyophilised with 20% w/v mannitol or 10% w/v dried milk since these lyoprotective agents demonstrated they are effective at maintaining cells viability. Nevertheless, these agents did not allow acetic acid production from ethanol. This study has demonstrated that glycerol (20% v/v) and mannitol (20% w/v) can be used as cryoprotective agents during freezing since they not only protect AAB and maintain bacteria viability but also help to preserve the functional properties, such as its ability to grow and produce acetic acid in alcoholic musts. / [ES] Las bacterias del ácido acético (BAA) pertenecen a la familia Acetobacteriaceae; están incluidas en el grupo de las ¿-Proteobacterias. Son microorganismos Gram-negativos, de forma elipsoidal o cilíndrica que pueden encontrarse aislados, en parejas o formando cadenas. Son móviles por flagelación polar o perítrica. Presentan actividad catalasa positiva, oxidasa negativa y no forman endosporas. Poseen metabolismo aeróbico estricto, con el oxígeno como aceptor final de electrones. Actualmente, la familia Acetobactereaceae está compuesta por 19 géneros y 72 especies. Es conocida la habilidad de las BAA para oxidar azúcares y alcoholes, obteniéndose como producto final una acumulación de ácidos orgánicos, capacidad que es aprovechada en la industria de alimentos para la elaboración de vinagres de vinos y de frutas. La presente Tesis Doctoral planteó el aislamiento e identificación de BAA a partir de la flora epifítica de arándanos y frutas cítricas cultivadas en la región de Salto Grande (Entre Ríos, Argentina), con el fin de encontrar las más adecuadas para ser utilizadas en procesos biotecnológicos, tales como el vinagre. Se aislaron 36 BAA a partir de estas frutas mediante técnicas de enriquecimiento y aislamiento en placas. Los caldos de enriquecimiento que contenían etanol y ácido acético permitieron recuperar el mayor número de BAA, ya que dichos componentes favorecen el crecimiento de las mismas. Del jugo fermentado de las frutas cítricas se obtuvo un mayor número de BAA respecto del jugo fresco o la cáscara, debido a la presencia de etanol, el que actuó como agente de selección para estos microorganismos alcohol-resistentes. Las pruebas bioquímicas permitieron diferenciar las bacterias a nivel de género. Se reconocieron 6 géneros: 13 aislados fueron identificados como Acetobacter, 5 como Gluconobacter, 7 como Asaia, 5 como Acidomonas, 1 como Gluconacetobacter y 5 como Saccharibacter. Posteriormente, se identificaron a nivel de especie aquellas bacterias (géneros Acetobacter, Gluconobacter y Gluconacetobacter) que podrían ser utilizadas para el desarrollo de un cultivo iniciador apto para la bioxidación de mostos alcohólicos obtenidos a partir de estos frutos, con el fin de obtener vinagres. Para esto, se emplearon técnicas moleculares, tales como PCR-RFLP del gen 16S y PCR-RFLP del espaciador intergénico 16S-23S. Con la primera, se identificaron las 16 BAA estudiadas. Los aislados C7, C8, A80, A160 y A180 fueron identificados como G. frateurii, los aislados C1, C2, C3, C4, C5, C6, A70 y A210 como A. pasteurianus, los aislados A50 y A140 como A. tropicalis y el aislado C9 como A. syzygii. Se estudió la dinámica de crecimiento y la habilidad de las BAA aisladas para producir ácido acético, con el fin de elegir las más aptas para la acetificación de mostos de frutas. El crecimiento se evaluó con distintas concentraciones de etanol y ácido acético. En cuanto a la habilidad para producir ácido acético, se ensayaron tres concentraciones de etanol, 4%, 6% y 8%, evidenciándose que los cultivos, identificados como A. pasteurianus (A210 y C1) A. syzygii (C9) y G. frateurii (A80) podrían ser utilizados como inóculo para la elaboración de vinagres, por poseer una alta capacidad de producción de ácido acético cuando la concentración de etanol es de 4% y 6% v/v. Las BAA podrían ser liofilizadas con manitol 20% p/v o leche en polvo 10% p/v, ya que estos lioprotectores demostraron ser efectivos para mantener la viabilidad de las células. Sin embargo, éstos no permitieron mantener la producción de ácido acético a partir de etanol. El presente estudio demostró que el glicerol (20% v/v) y el manitol (20% p/v) pueden ser utilizados como crioprotectores en el proceso de congelación, ya que no sólo protegen a las BAA, manteniendo su viabilidad, sino que también ayudan a conservar las propiedades funcionales de las mismas, tales como su capacidad de crecer y producir ácido / [CAT] Els bacteris de l'àcid acètic (BAA) pentanyen a la família Acetobacteriaceae i estan incloses en el grup dels ¿-Proteobacteris. Són microorganismes gram-negatius, de forma elipsoidal p cilíndrica que pot trobar-se aïllada, emparellada o formant cadenes. Són mòbils per flagel¿lació polar o perítrica. Presenten activitat catalasa positiva, oxidasa negativa i no formen endospores. Posseeixen metabolisme aeròbic estricte, amb l'oxigen com aceptor final d'electrons. Actualment, la familia Acetobactereaceae està composta per 19 gèneres i 72 espècies. Es coneguda l'habilitat dels BAA per a oxidar sucres i alcohols, obtenint-se com a producte final una acumulació d'àcids orgànics, capacitat aprofitada en la industria dels aliments per a l'elaboració de vinagres de vins i fruites. La present Tesi Doctoral planteja l'aïllament i identificació dels BAA a partir de la flora epifítica dels nabius i cítrics cultivats en la regió de Salto Grande (Entre Ríos, Argentina), amb la finalitat de trobar les més adequades per ser utilitzades en processos biotecnològics, tals com el vinagre. S'aïllaren 36 BAA a partir d'aquestes frutes mitjançant tècniques d'enriquiment i aïllament en plaques. Els brous d'enriquiment que contenien E i AA permeteren recuperar el major numero de BAA, ja que tals components afavorien el seu creixement. Del suc fermentat dels citrics es va obtenir un major nombre de BAA respecte del suc fresc o la pell, degut a la presència de E, que actuà com agent de selecció per a aquestos microorganismes alcohol-resistents. Es reconegueren 6 gèneres: 13 aïllats foren identificats com Acetobacter, 5 com Gluconobacter, 7 com Asaia, 5 com Acidomonas, 1 com Gluconacetobacter i 5 com Saccharibacter. Posteriorment, s'identificaren a nivell d'espècie aquells bacteris que podríen ser utilitzats per al desenvolupament d'un cultiu indicadorstarter apte per a la biooxidació de mostos alcohòlics obtinguts a partir d'aquestos fruïts, amb la finalitat d'obtenir vinagres de fruites (gèneres Acetobacter, Gluconobacter i Gluconacetobacter). Amb aquesta finalitat, s'utilitzaren tècniques moleculars, com PCR-RFLP del gen 16S i PCR-RFLP de l'espaciador intergènic 16S-23S. Amb la primera, s'identificaren els 16 BAA estudiats. La tècnica PCR-RFLP del 16S-23S va confirmar les identificacions realitzades amb el 16S, tanmateix C1 va mostrar un patró de restricció que no es corresponia amb la primera identificació realitzada. Aquesta discrepància fou resolta per la seqüenciació parcial de gen 16S, que va confirmar el resultats obtinguts per PCR-RFLP. Els aïllats C7, C8, A80, A160 i A180 foren identificats com G. Frateurii, els aïllats C1, C2, C3, C4, C5, C6, A70 i A210 com A. pasteurianus,els aïllats A50 i A140 com A.tropicalis i l'aïllat C9 com A. syzygii. S'estudià la dinàmica de creixement i la habilitat per produirAA dels BAA aïllades, amb la finalitat de triar les més aptes per a l'acetificació de mostos de frutes. El creixement s'evaluà amb diferents concentracions de E i AA. Quan a l'habilitat per a produir AA, s'assajaren tres concentracions d'etanol, 4%, 6% i 8% v/v, evidencianse que els cultius , identificats com A. pasteurianus (A210 i C1) A. syzygii (C9) i G. frateurii (A80) podrien ser utilitzats com inòcul per a l'elaboració de vinagres, per poseïr una alta capacitat de producció de AA quan la concentraició de E és de 4% i 6% v/v. Els BAA podrien ser liofilitzats amb manitol 20% p/v o llet en pols 10% p/v, ja que aquestos lioprotectors demostraren ser efectius per a mantenir la viabilitat de les cèl¿lules. Tanmateix, aquestos no permeteren mantenir la producció de AA a partir de E. El present estudi va demostrar que el glicerol (20% v/v) i el manitol (20% p/v) poden ser utilitzats com crioprotectors en el procés de congelació, ja que no sols protegeixen els BAA, mantenint la seua viabilitat, sinò que també ajuda a conservar les seues propietats funcionals, tals com la se / Gerard, LM. (2015). Caracterización de bacterias del ácido acético destinadas a la producción de vinagres de frutas [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/59401 / TESIS

This thesis is concerned with the identification of the yeasts belonging to Saccharomyces species, which is participating in the particular fermentation phases of the white wine. The rapidity that we are able to identify the yeast strains is the significant factor for appreciation the fermantation proces quality as well as final product quality – the quality of wine. The molecular biology method developing is gradually limiting the using of traditional identification methods mainly because of their time intensity. In this thesis was used molecular method PCR-RFLP as an implement to quick and accurate identification of particular strains of yeasts. The specific DNA section was amplified by the help of PCR and consequently amenabled to the restrict analysis. The restrict endonukleas fissiled DNA to fragments specified for the certain strain of yeast. The fragments were detected by horizontal electrophoresis. To compare the fragments with type yeast fragments made us possible to identify the trast strain and its taxonomy classification. The literature search contains the basic information about the yeasts and their using, the information about wine making as well as the PCR-RFLP method principle.

This thesis deals with identification of the wine yeasts by applying the PCR-RFLP method. The identification and characteristic of the yeasts has gone through substantial changes in recent years. There have been introduced new methods of taxonomic classifying based on the molecular methods, which are oriented to easy and fast identification. One of these methods is the PCR-RFLP method. The amplification of the 5•8S-ITS rDNA sequence by the polymerase chain reaction with use of the primers ITS1 and ITS4 leads to the amplification of the specific sequence of DNA. Such multiplied DNA is after repurifying by the ethanol and drying submitted to the restriction analysis. With use of the restriction endonuklases DNA is chopped into the specific segments typical for the particular genus. The chopped fragments can be separated in the electric field in the agarose gel and subsequently evaluated. In this thesis together 63 type yeasts were used. These yeasts were analysed by applying of the seven restriction endonuklases – HaeIII, HhaI, HinfI, HpaII, TaqI, AluI a MseI. The final image of type yeasts splitting was compared to the results of splitting of already identified wine yeasts and these yeasts were subsequently taxonomically classified. Evaluation of genetic similarity was conducted by program BioNumerics and as the results the dendrograms that were created with use of Jaccard‘s coefficients are obtained.