Neozygites floridana is an important natural enemy of Tetranychus urticae in many crops, including strawberry. In this crop there has been some changes in the production system from open field to low tunnel. This new system presents benefits to strawberry production, moreover this system increases problems with spider mites and some fungal plant pathogens; consequently increasing the use of pesticides. In order to integrate N. floridana in the management of spider mites in strawberry there are still many important aspects to be revealed on the biology, epizootiology of the fungus and the effect of crop systems on its prevalence in the field. For this, three studies were performed, being the first one about the effect of strawberry production systems (low tunnel and open field) and pesticide application on population dynamics of two-spotted spider mite T. urticae, its natural enemies (N. floridana and predatory mites), and plant diseases, the experiment was performed in Inconfidentes-MG. The population of T. urticae attained the highest level in treatments with chemical control of pest, independently of the crop system, low tunnel or open field. The fungus N. floridana was observed late in both crop seasons, in treatments with and without inoculative releases, and has suggested that it might be a natural occurrence of the fungus. The highest prevalence of the fungus were observed in treatments with high densities of T. urticae. The predatory mites occurred naturally during the two crop seasons, and the most abundant species was Neoseiulus anonymus (Chant) (57.5%). The incidence of fungal plant diseases are greater in the open field than in low tunnel, however it seems that the crop system do not affect the new disease \"vermelhao\". The second study aimed to reveal the abiotic and biotic factors involved in the regulation of resting spores formation in the hosts T. urticae and T. evansi. The Brazilian isolate ESALQ1420 produced a large number of resting spores (51.5%) in T. urticae at a temperature 11 °C, photoperiod of 10L:14D, light intensity of 42-46 (?mol m-2s-1), on non-senescent plants. Small percentages of mites with resting spores (0-5 %) were found for the Norwegian isolate NCRI271/04 under the conditions tested, and very low percentages of resting spores (up to 1%) were observed in T. evansi infected by the Brazilian isolate ESALQ1421.The third study was therefore conducted phylogenetic analysis of isolates of N. floridana and develops efficient real-time PCR-based analysis for detection and quantification of N. floridana propagules from the soil. The phylogenic three showed that Neozygites is a fungal group far from other Entomophthoromycota and that N. floridana present high intra-specific variability. This data indicates that the isolates from different Tetranychus species may be in fact different Neozygites species, not described yet. We developed a real-time PCR probe for detection and quantification of N. floridana in the soil. This molecular probe was successful in quantifying N. floridana at relatively high DNA concentrations seeded into the soil. / Neozygites floridana é um importante inimigo natural de Tetranychus urticae em diversas culturas, incluindo morango. Nesta cultura tem havido uma mudança no sistema de produção, passando de cultivo em campo aberto para túnel baixo. Este novo sistema apresenta benefícios na produção de morango, no entanto pode aumentar problemas com ácaros fitófagos e algumas doenças fúngicas, consequentemente aumentando o uso de agrotóxicos. Para a integração de N. floridana no manejo de ácaros fitófagos em morango, existem ainda muitos aspectos importantes a serem conhecidos sobre a biologia, epizootiologia do fungo e efeito dos sistemas de cultivo sobre a sua prevalência em campo. Para isso três estudos foram realizados, sendo o primeiro estudo sobre o efeito de sistemas de produção de morango (túnel baixo e campo aberto) e da aplicação de agrotóxicos sobre a dinâmica populacional do acaro rajado T. urticae, seus inimigos naturais (N. floridana e ácaros predadores) e doenças de plantas, o experimento foi realizado em Inconfidentes-MG. A população de T. urticae atingiu altos níveis em tratamentos com controle químico de pragas, independente do sistema de cultivo, túnel baixo ou campo aberto. O fungo N. floridana foi observado no final de ambos os ciclos de cultivos, nos tratamentos com e sem liberações inoculativas, sugerindo que houve incidência natural do fungo. As maiores prevalências do fungo foram observadas em tratamentos com altas densidades de T. urticae. Os ácaros predadores ocorreram naturalmente durante os dois ciclos de cultivo, sendo Neoseiulus anonymus (Chant) a espécie mais abundante (57.51%). A incidência de doenças fúngicas foi maior em campo aberto do que em túnel baixo, no entanto parece que o sistema de cultivo não afetou a incidência da nova doença vermelhão. No segundo estudo objetivou-se conhecer os fatores abióticos e bióticos envolvidos na formação de esporos de resistência nos hospedeiros T. urticae e Tetranychus evansi. O isolado brasileiro ESALQ1420 produziu um grande número de esporos de resistência (51.5%) em T. urticae, a uma temperatura de 11 ° C, fotoperíodo de 10F:14E, intensidade de luz de 42-46 (?mol m-2s-1), em plantas senescentes. Pequenas percentagens de ácaros com esporos de resistência (0-5%) foram encontrados para o isolado Norueguês NCRI271/04 sob as condições testadas, e muito baixas percentagens de esporos de resistência (ate 1%) foram observados em T. evansi infectados pelo isolado brasileiro ESALQ1421. O terceiro estudo objetivou analisar filogeneticamente os isolados de Neozygites patogênicos a ácaros, e desenvolver ferramenta utilizando PCR em tempo real para a detecção e quantificação de propágulos de N. floridana a partir do solo. A arvore filogenética mostrou que Neozygites e um grupo de fungos distante dos demais Entomophthoromycota, e que estes fungos apresentaram alta variabilidade intraespecífica entre isolados de N. floridana. Estes resultados indicam que isolados provenientes de diferentes espécies hospedeiras do gênero Tetranychus podem ser de fato diferentes espécies de Neozygites não descritas ainda. Desenvolveu-se uma sonda molecular para detecção e quantificação de N. floridana no solo utilizando PCR em tempo real. Esta sonda foi eficiente na quantificação de N. floridana, em concentrações relativamente altas de DNA inoculado no solo.

As micorrizas arbusculares (MAs) são simbiontes mutualísticas entre alguns fungos do solo e raízes de plantas de notória importância tanto em ecossistemas naturais quanto agrícolas, pois contribuem para o aumento da absorção de nutrientes do solo e sua transferência para as plantas. As MAs podem promover o crescimento de várias plantas, dentre elas a cana-de-açúcar. Assim, fatores que restringem o desenvolvimento das MAs podem também restringir a produção das culturas agrícolas. Muito embora os herbicidas utilizados em sistemas agrícolas possam contribuir para a alteração das estruturas das comunidades de microrganismos no solo e o desenvolvimento de simbioses, pouco se sabe sobre seu efeito sobre fungos micorrízicos arbusculares (FMAs). O presente estudo teve por objetivo determinar o efeito dos herbicidas Ametrina e Imazapique na germinação de esporos de Glomus clarum e desenvolvimento de MA em cana-de-açúcar, bem como as alterações no transcritoma de raízes de cana-de-açúcar colonizadas por G. clarum e cultivada em solo tratado com Imazapique. Ametrina não teve efeito significativo na germinação de esporos e colonização intrarradicular, nas doses avaliadas. Já, o aumento da concentração de Imazapique promoveu uma diminuição na germinação de esporos e aumento de colonização intrarradicular. Embora tenha sido observada maior taxa de colonização intrarradicular em plantas tratadas com Imazapique, a presença do fungo parece não ter aliviado os efeitos deletérios promovidos pelo uso do herbicida nas condições estudadas. Através de hibridização em macroarranjo de cDNA, foi possível detectar vários genes com expressão diferencial estatisticamente significativa em raízes micorrizadas e não- micorrizadas cultivadas em solo tratado com herbicida. Dentre as proteínas codificadas pelos genes com expressão induzida em raízes micorrizadas, em relação ao controle não-inoculado, se destacam um receptor de glutamato, uma metalotioneína, uma ATP sintase, e várias com função desconhecida. Já, dentre os genes com expressão suprimida estão: a proteína WALI7, uma proteína de catabolismo dependente de ubiquitina, uma tubulina e uma remorina. Nas raízes não-inoculadas e cultivadas em solo com Imazapique as proteínas codificadas pelos genes com expressão induzida são: um receptor de glutamato e uma proteína tipo lípase; e dentre as que foram suprimida está a proteína transcritase reversa. O Imazapique altera a expressão de vários genes modulados pela MA, dentre as proteínas codificadas por esses genes estão: uma ATP sintase, uma proteína de catabolismo dependente de ubiquitina, uma poliubiquitina, uma tubulina, uma quinase de histidina e várias com função desconhecida. Os resultados indicam que a formação MA altera o programa genético das raízes de cana-de-açúcar e que o herbicida Imazapique altera a expressão dos genes relacionados à simbiose de cana-de-açúcar com G. clarum. / The arbuscular mycorrhizae (AM) are symbiotic root-fungus associations that play a key role in natural and agricultural ecosystems, through the enhancement of nutrients absorption in the soil and its transference into the plants. The AM may promote the growth of many plants, among them are the sugarcane. Even thought the herbicide utilized in the agriculture system may contribute to soil microbial community alteration and the symbioses development, limited is know about the herbicide effects on arbuscular mycorrhizal fungi (AMF) and on AM formation. The aim of the present work was to evaluate the Ametrine and Imazapic effects in the spores germination of Glomus clarum and AM development in sugarcane, as well transcriptome alterations of sugarcane roots colonized with G. clarum and growing in soil treat with Imazapic. It was not detected significant effect on the spores germination and intraradical colonization in the evaluated doses of Ametrine. Whereas, the enhance of Imazapic concentration promoted a decrease in the spores germination and increase of intraradical colonization. Although the higher intraradical colonization has been observed in the plants treat with Imazapic, the presence of the fungi seems not alleviated the damage effect promoted by the use of the herbicide, in these studies conditions. By macro array cDNA hybridizations, it was possible to detect several genes with differential expression statistically significant in AM plants, and plants not inoculated growing in soil treated with herbicide. Among the proteins codified by genes with induced expression in AM roots, compared to the control no-inoculated, are: a putative ionotropic glutamate receptor, a metallothionein-like protein, a ATP synthase and many others with unknown function. Whereas, among the proteins codified by genes with suppressed expression are: Wali7 protein, a ubiquitin-dependent protein catabolism, a tubulin alpha-1 chain and a remorin. In the no-inoculated roots growing in the soil treat with Imazapic the proteins codified by genes with induced expression are: a putative ionotropic glutamate receptor and a lipase-like; and the reverse transcriptase like protein was suppressed. The Imazapic treatment alter several gene expression modulated by AM, among the proteins codified by those genes are: a ATP synthase, a ubiquitin-dependent protein catabolism, polyubiquitin, a tubulin alpha-1 chain, a signal transduction histidine kinase and many others with unknown function. The results indicate the AM formation alter the sugarcane roots genetic program and that the Imazapic herbicide alter the gene expression related to symbioses between sugarcane with G. clarum.

Com este estudo, objetivou-se avaliar o efeito de estirpes de Bacillus thuringiensis (Bt) na degradabilidade e digestibilidade da dieta, emissão de gases, microbiota ruminal, parâmetros sanguíneos e desempenho de ovinos. O estudo foi dividido em 2 experimentos: ensaio in vitro de produção de gases para a avaliação de 6 diferentes estirpes de Bt (907, 1192, 2036, 2493, 2496 e S1185) e ensaio in vivo com a estirpe selecionada 2036 para investigação de possíveis interferências na digestão e saúde dos animais. A simulação do ambiente ruminal foi realizada em garrafas de vidro incubadas a 39 oC por 24 h. O delineamento foi o inteiramente casualizado com 7 tratamentos (Sem Bt (controle), Bt 907, Bt 1192, Bt 2036, Bt 2493, Bt 2496 e Bt S1185) com 4 repetições em duplicata. O processo fermentativo foi avaliado pelos resultados de matéria seca degradada (MSD), matéria orgânica degradada (MOD), produção líquida de gases totais, produção líquida de metano e eficiência da conversão de metano. Produtos da fermentação (pH, nitrogênio amoniacal (N-NH3) e ácidos graxos de cadeia curta (AGCC)) e micro-organismos ruminais (Fibrobacter succinogenes, Ruminococcus flavefaciens, fungos anaeróbicos, arqueas metanogênicas e protozoários) também foram estudados. Apenas a estirpe Bt 907 reduziu a MSD e MOD em relação ao controle, com manutenção da população de F. succinogenes, pois as demais estirpes reduziram a população desta bactéria. No ensaio in vivo 20 cordeiros Santa Inês com 3 meses de idade e 18 ± 3,5 kg PV foram utilizados e divididos em 2 grupos: 10 animais tratados com 2,5x106 esporos de Bt 2036 por kg PV/d e 10 animais não tratados (controle). Estes foram alojados em baias individuais em delineamento inteiramente casualizado e receberam dieta composta de feno de capim Tifton-85 (Cynodon spp.) ad libitum e 300 g/animal/d de concentrado, que foi ajustado de acordo com as exigências de crescimento. O período experimental in vivo teve duração de 63 dias, dos quais 53 compreendeu o teste de desempenho dos animais, com aferição do consumo 3 vezes na semana e pesagem quinzenal, e os 10 dias subsequentes destinou-se aos ensaios de digestibilidade aparente, balanço de nitrogênio, síntese de proteína microbiana e emissão de metano entérico. Durante todo o experimento, coletas se sangue foram realizadas quinzenalmente a fim de avaliar os parâmetros hematológicos (hemácias, hemoglobina, hematócrito e leucócitos) e bioquímicos (glicose, proteínas totais, albumina, aspartato aminotransferase, ureia e creatinina) dos animais para o diagnóstico de possível intoxicação. Características da fermentação ruminal também foram investigadas em 3 momentos (início, meio e fim do experimento) sobre as variáveis pH, N-NH3, AGCC, abundância relativa das espécies F. succinogenes, R. flavefaciens, populações de arqueas metanogênicas, fungos anaeróbicos e contagem de protozoários, por meio de coletas de líquido ruminal. Não houve influência da estirpe sobre as variáveis estudadas. Conclui-se que na avaliação in vitro apenas a estirpe Bt 907 reduziu a MSD e MOD com manutenção da população F. succinogenes e no experimento in vivo a inclusão de esporos de Bt 2036 na dieta não afetou de forma negativa o desempenho e nem a saúde dos ovinos / The objective of present study was to evaluate the effect of Bacillus thuringiensis (Bt) strains on degradability and digestibility of the diet, gas production, ruminal fermentation, blood parameters and performance in sheep. The study was divided into 2 experiments: in vitro gas production to evaluate six different Bt strains (907, 1192, 2036, 2493, 2496 and S1185) and in vivo assay with the selected strain (Bt 2036) to investigate possible interference in digestion and health of animals. The rumen simulation was performed in glass bottles incubated at 39 °C for 24 h. A completely randomized design with 7 treatments (No Bt (control), Bt 907, Bt 1192, Bt 2036, Bt 2493, Bt 2496 and Bt S1185) was used, with 4 replications in duplicate. The fermentation process was evaluated using dry matter degradability (DMD), organic matter degradability (OMD), net gas production, methane output and conversion efficiency. Fermentation products (pH, ammonia nitrogen (N-NH3), short chain fatty acids (SCFA)) and ruminal microorganisms (Fibrobacter succinogenes, Ruminococcus flavefaciens, anaerobic fungi, methanogenic archaea and protozoa) were also studied. Only strain Bt 907 reduced DMD and OMD in relation to the control and F. succinogenes populations were maintained, whereas other strains of this bacterium population were reduced. An in vivo assay using 20 Santa Ines lambs at 3 months of age and 18 ± 3.5 kg BW was carried out. These animals were divided into 2 groups: 10 animals treated with 2.5x106 spores of Bt 2036 per kg BW/d and 10 untreated animals (control). These were housed in individual pens in a completely randomized design and were fed a diet of Tifton-85 (Cynodon spp.) hay ad libitum and 300 g/animal/d of concentrate, which was adjusted according to animal growth. The in vivo experimental period lasted 63 days, of which 53 included the performance test of the animals, with measurement of feed consumption three times a week and fortnightly weighing. The final 10 days was devoted to tests of digestibility, nitrogen balance, microbial protein synthesis and enteric methane emission. Throughout the experiment, blood was collected fortnightly to assess hematological parameters (erythrocytes, hemoglobin, packed cell volume and leukocytes) and biochemical profiles such as (Glucose, total protein, albumin, aspartate aminotransferase, urea and creatinine) for the diagnosis of possible intoxication. Ruminal fermentation characteristics were investigated at three times (Initial, middle and end of the experiment) using pH, N-NH3, SCFA, microorganism population size (such as F. succinogenes, R. flavefaciens, methanogenic archaea, anaerobic fungi and protozoa) through ruminal fluid collections. There was no influence of Bt 2036 on ruminal fermentation characteristics. It is concluded that, for the in vitro evaluation, only strain Bt 907 reduced DMD and OMD, with F. succinogenes population being maintained and for the in vivo studies, the inclusion of Bt 2036 spores in the diet did not negatively affect health and performance of lambs

OBJETIVOS: Estudar o ambiente interno em escolas públicas situadas em duas cidades (Atibaia e São Paulo), com diferentes níveis de poluição atmosférica, mediante a identificação e quantificação dos principais alérgenos presentes na poeira e no ar das salas de aula dessas escolas, assim como a avaliação da existência de padrão sazonal para cada um dos alérgenos estudados. MATERIAIS E MÉTODOS: As amostras de poeira foram coletadas de uma única sala de aula em cada escola, uma vez por mês durante um ano. A cada visita foram medidas a temperatura e umidade relativa do ar. A aspiração da poeira foi realizada em 8 pontos da sala, sendo 2 próximos à janela, 2 junto à porta, 2 junto à lousa e 2 no centro. Uma das amostras aspirada de cada local foi analisada quanto à sua composição, através de leitura por ELISA, utilizando-se anticorpos monoclonais (Indoor Biotechnologies-USA), para dosagem dos seguintes alérgenos: Der p 1, Der f 1, Blo t 5, Fel d 1, Can f 1, Asp f 1, Alt a 1, Bla g 1 e Bla g 2. A outra amostra foi semeada em agar Sabouraud para cultura para fungos. As amostras de ar foram coletadas por 20 minutos com o aparelho de Andersen N-6, um impactador de 6 estágios, colocado no centro da sala de aula. Os gêneros dos fungos foram identificados pela observação macroscópica das xi colônias e pelas características microscópicas das hifas, determinadas pela técnica de Ridell. RESULTADOS: Os resultados mostraram baixos níveis (< 2ug/g) de alérgenos de ácaros (Der p 1, Der f 1 e Blo t 5) na maioria das amostras de poeira dos pisos das escolas. Também foram encontrados baixos níveis (< 1ug/g) dos alérgenos animais (Fel d 1 e Can f 1), sendo que maiores níveis de Can f 1 foram detectados nas amostras da escola de São Paulo. Os alérgenos de barata não foram detectados nas duas escolas. As maiores quantidades de alérgenos de ácaros ocorreram durante a primavera e as menores no outono. Em Atibaia, verificaram-se maiores níveis de alérgenos de ácaros e esporos de fungos junto à lousa. Foram identificados 19 gêneros de fungos nas amostras de poeira e 25 gêneros nas amostras de ar das duas escolas. Destacaram-se Cladosporium nas amostras coletadas com aparelho de Andersen e Trichoderma nas amostras aspiradas. Considerando-se o total de esporos de fungos obtidos com aparelho de Andersen, não houve diferença estatisticamente significante entre as escolas de Atibaia e São Paulo, assim como não houve diferença entre as estações do ano. Em relação às amostras de poeira coletadas com aspirador, São Paulo apresentou os maiores níveis de UFC/g de poeira. CONCLUSÃO: Os pisos e o ar das escolas estaduais estudadas parecem não representar fonte importante de exposição à alérgenos de ácaros, fungos, cães, gatos e baratas, independente da presença de poluição atmosférica / Objective: To study indoor environment in public schools located in two cities (Atibaia and São Paulo) with different degrees of air pollution, identifying and measuring the most important allergens in classroom dust and air samples and evaluating the existence of seasonal variation for each allergen. Methods: Dust samples were obtained from a single classroom in each school, once a month during a year. During each visit temperature and relative humidity were measured. Floor dust was collected from eight different sites: 2 near the door, 2 near the window, 2 near the blackboard and 2 in the middle of the room. One sample collected from each site was analyzed by using mAb-based ELISAs (Indoor Biotechnologies-USA) for Der p 1, Der f 1, Blo t 5, Asp f1, Alt a 1, Fel d 1, Can f 1, Bla g 1 and Bla g 2. The other samples were plated on Sabouraud dextrose agar. Air samples were collected for 20 minutes with a N-6 Andersen sampler, a six-stage impactor, located in the middle of the classroom. Fungal genera were identified by macroscopic observation of the colonies and by the microscopic characteristics of the sporulating hyphae determined by the Ridell technique. Results: The results showed low levels (<2ug/g) of mite allergens (Der p 1, Der f 1 and Blo t 5) in most dust samples collected from the classroom\'s floor. Low levels (<1ug/g) of animal allergens (Fel d 1 and Can f 1) were found too, and higher levels of Can f 1 were detected in dust samples from São Paulo. Cockroach allergens (Bla g 1 and Bla g 2) were undetectable in all samples. Higher levels of mite allergens occurred during spring and lower during autumn. Higher levels of mite allergens and fungal spores were found near the blackboard in Atibaia. A total of 19 different fungal genera were distinguished in dust samples, while 25 genera were identified in air samples. The most frequently isolated fungal genera in Andersen samples was Cladosporium and in dust samples was Trichoderma. Considering total mold spores obtained with Andersen sampler, there was no statistically significant difference between São Paulo and Atibaia schools, neither between the seasons of the year. In relation to dust samples collected with vaccum cleaner, the school from São Paulo presented higher levels of UFC/g of dust. Conclusion: The floor and air from these two elementary schools in São Paulo seems not to represent an important source of exposure to allergens from mites, molds, cats, dogs and cockroach, independent on the presence of air pollution.

As micorrizas arbusculares (MAs) são simbiontes mutualísticas entre alguns fungos do solo e raízes de plantas de notória importância tanto em ecossistemas naturais quanto agrícolas, pois contribuem para o aumento da absorção de nutrientes do solo e sua transferência para as plantas. As MAs podem promover o crescimento de várias plantas, dentre elas a cana-de-açúcar. Assim, fatores que restringem o desenvolvimento das MAs podem também restringir a produção das culturas agrícolas. Muito embora os herbicidas utilizados em sistemas agrícolas possam contribuir para a alteração das estruturas das comunidades de microrganismos no solo e o desenvolvimento de simbioses, pouco se sabe sobre seu efeito sobre fungos micorrízicos arbusculares (FMAs). O presente estudo teve por objetivo determinar o efeito dos herbicidas Ametrina e Imazapique na germinação de esporos de Glomus clarum e desenvolvimento de MA em cana-de-açúcar, bem como as alterações no transcritoma de raízes de cana-de-açúcar colonizadas por G. clarum e cultivada em solo tratado com Imazapique. Ametrina não teve efeito significativo na germinação de esporos e colonização intrarradicular, nas doses avaliadas. Já, o aumento da concentração de Imazapique promoveu uma diminuição na germinação de esporos e aumento de colonização intrarradicular. Embora tenha sido observada maior taxa de colonização intrarradicular em plantas tratadas com Imazapique, a presença do fungo parece não ter aliviado os efeitos deletérios promovidos pelo uso do herbicida nas condições estudadas. Através de hibridização em macroarranjo de cDNA, foi possível detectar vários genes com expressão diferencial estatisticamente significativa em raízes micorrizadas e não- micorrizadas cultivadas em solo tratado com herbicida. Dentre as proteínas codificadas pelos genes com expressão induzida em raízes micorrizadas, em relação ao controle não-inoculado, se destacam um receptor de glutamato, uma metalotioneína, uma ATP sintase, e várias com função desconhecida. Já, dentre os genes com expressão suprimida estão: a proteína WALI7, uma proteína de catabolismo dependente de ubiquitina, uma tubulina e uma remorina. Nas raízes não-inoculadas e cultivadas em solo com Imazapique as proteínas codificadas pelos genes com expressão induzida são: um receptor de glutamato e uma proteína tipo lípase; e dentre as que foram suprimida está a proteína transcritase reversa. O Imazapique altera a expressão de vários genes modulados pela MA, dentre as proteínas codificadas por esses genes estão: uma ATP sintase, uma proteína de catabolismo dependente de ubiquitina, uma poliubiquitina, uma tubulina, uma quinase de histidina e várias com função desconhecida. Os resultados indicam que a formação MA altera o programa genético das raízes de cana-de-açúcar e que o herbicida Imazapique altera a expressão dos genes relacionados à simbiose de cana-de-açúcar com G. clarum. / The arbuscular mycorrhizae (AM) are symbiotic root-fungus associations that play a key role in natural and agricultural ecosystems, through the enhancement of nutrients absorption in the soil and its transference into the plants. The AM may promote the growth of many plants, among them are the sugarcane. Even thought the herbicide utilized in the agriculture system may contribute to soil microbial community alteration and the symbioses development, limited is know about the herbicide effects on arbuscular mycorrhizal fungi (AMF) and on AM formation. The aim of the present work was to evaluate the Ametrine and Imazapic effects in the spores germination of Glomus clarum and AM development in sugarcane, as well transcriptome alterations of sugarcane roots colonized with G. clarum and growing in soil treat with Imazapic. It was not detected significant effect on the spores germination and intraradical colonization in the evaluated doses of Ametrine. Whereas, the enhance of Imazapic concentration promoted a decrease in the spores germination and increase of intraradical colonization. Although the higher intraradical colonization has been observed in the plants treat with Imazapic, the presence of the fungi seems not alleviated the damage effect promoted by the use of the herbicide, in these studies conditions. By macro array cDNA hybridizations, it was possible to detect several genes with differential expression statistically significant in AM plants, and plants not inoculated growing in soil treated with herbicide. Among the proteins codified by genes with induced expression in AM roots, compared to the control no-inoculated, are: a putative ionotropic glutamate receptor, a metallothionein-like protein, a ATP synthase and many others with unknown function. Whereas, among the proteins codified by genes with suppressed expression are: Wali7 protein, a ubiquitin-dependent protein catabolism, a tubulin alpha-1 chain and a remorin. In the no-inoculated roots growing in the soil treat with Imazapic the proteins codified by genes with induced expression are: a putative ionotropic glutamate receptor and a lipase-like; and the reverse transcriptase like protein was suppressed. The Imazapic treatment alter several gene expression modulated by AM, among the proteins codified by those genes are: a ATP synthase, a ubiquitin-dependent protein catabolism, polyubiquitin, a tubulin alpha-1 chain, a signal transduction histidine kinase and many others with unknown function. The results indicate the AM formation alter the sugarcane roots genetic program and that the Imazapic herbicide alter the gene expression related to symbioses between sugarcane with G. clarum.

Ziel der vorliegenden Arbeit war zum einen die Ermittlung des im Grünfutter notwen-digen Mindest-Nitratgehaltes zur Erzielung buttersäurefreier Silagen, wobei TS-Gehalt und Z/PK-Quotient des Ausgangsmaterials sowie dessen Belastungsgrad mit Clostridiensporen berücksichtigt wurden. Dazu wurden zwei mehrfaktorielle Laborsi-lierversuche durchgeführt, bei denen eine Variationsreihe des Nitratgehaltes (0,01...0,3 % NO3-N in TS) mit Variationsreihen des TS-Gehaltes (ca. 14...40 %) und des Z/PK-Quotienten (1,5...3,1) systematisch kombiniert wurden. Jede Wertekombi-nation wurde sowohl mit sauber geerntetem Grünfutter als auch mit Clostridienspo-ren kontaminiertem Grünfutter geprüft. Die Silagen wurden nach 180 Tagen Lage-rungsdauer untersucht. Zum anderen wurde die Dynamik der Clostridienentwicklung im Gärungsverlauf in Abhängigkeit von TS-Gehalt, Säuerungsintensität und Nitratge-halt geprüft. Jede Stufe des TS-Gehaltes (20, 30, 40 und 50 %) wurde mit Zusätzen von Nitrat, Milchsäurebakterien (MSB) bzw. MSB + Glucose und MSB + Nitrat ange-setzt. Das Ausgangsmaterial (Dac. glomerata, nitratfrei) war durchgehend mit Clostridiensporen kontaminiert. Die Untersuchung der Silagen erfolgte 3, 7, 14, 28, 56 und 180 Tage nach dem Ansatz. Der Konservierungserfolg bei der Silierung hängt nicht nur vom TS-Gehalt und Z/PK-Quotienten sondern auch vom Nitratgehalt und Clostridiensporenbesatz des Aus-gangsmaterials ab. Bei sehr niedrigem Nitratgehalt des Grünfutters liegt ein erhöhtes Risiko für die Entstehung von Buttersäure in der Silage vor, auch bei dem als leicht vergärbar geltendem Grünfutter (VK > 45) und auch dann, wenn es im sauber geern-tetem Zustand einsiliert worden ist. Bei weiter erhöhten VK-Werten, > 45 (durch Er-höhung des TS-Gehaltes und/oder Z/PK-Quotienten), wird die Höhe der Buttersäure-gehalte zwar eingeschränkt. Zur sicheren Ausschaltung von Buttersäuregärung ist jedoch auch hier ein gewisser Nitratgehalt notwendig. Bei der Silierung nitratfrei-en/nitratarmen Grünfutters nimmt das Fehlgärungsrisiko mit dem Belastungsgrad an Clostridiensporen zu. Der notwendige Mindest-Nitratgehalt (MNG) hängt sowohl vom VK-Wert als auch vom Kontaminationsgrad des Grünfutters mit Clostridiensporen ab. Er ist um so niedriger, je höher der VK-Wert und geringer der Kontaminationsgrad ist, und umgekehrt. (MNG (% NO3-N in TS) für sporenarmes Grünfutter = 0,24 - 0,0035 . VK MNG (% NO3-N in TS) für sporenreiches Grünfutter = 0,20 - 0,0021 . VK) Hohe Clostridiensporengehalte lagen vor allem in buttersäurehaltigen Silagen vor und insbesondere dann, wenn das Grünfutter sehr geringe Nitratgehalte aufwies. Zwischen der Höhe der Buttersäuregehalte und dem Clostridiensporengehalt besteht jedoch kein direkter Zusammenhang. Erhöhung des TS-Gehaltes bewirkt eine Ein-schränkung der Clostridienentwicklung. Ein Rückgang des Sporengehaltes im Ver-gleich zum Ausgangsmaterial (nitratfrei) lag aber erst bei einem TS-Gehalt von etwa 50 % vor. Durch Zusätze von MSB sowie MSB + Glucose konnte die Milchsäuregä-rung deutlich intensiviert werden. Ein sehr geringer pH-Wert war schon am 3. oder 7. Lagerungstag erreicht. Buttersäuregärung war aber erst bei TS-Gehalten > 40 % ausgeschaltet. Eine Einschränkung der Sporenbildung lag ebenfalls erst bei TS-Gehalten über 40 % vor. Bei Nitratzusatz blieben die Silagen aller TS-Stufen bis zur Auslagerung buttersäurefrei. Die Sporengehalte gingen in allen TS-Stufen während des Gärungsverlaufes kontinuierlich zurück. Bei steigenden TS-Gehalten war der Rückgang der Sporengehalte verlangsamt. Durch die Kombination von MSB und Nit-rat konnte nicht nur ein sicherer Erfolg bei der Unterbindung von Buttersäurebildung und Laktatabbau sondern auch eine starke Verminderung der Sporengehalte erreicht werden. / The first aim of this work was to determine the minimum content of nitrate (MCN) which is required to get silage free of butyric acid. For it, two multi-factorial experi-ments with orchardgrass were carried out under laboratory condition. In this experi-ments, nitrate content (0.01 ... 0.3 % N in DM) was systematically combined with staggered levels of the dry matter (DM) content (14 ... 40 %) and of the ratio of water-soluble carbohydrate to buffering capacity (WSC/BC: 1.5 ... 3.1). All variants were tested with forage without or with addition of clostridial spores. The silages were ana-lysed after 180 days of incubation. The second aim of this study was to explain the dynamic of clostridial development during ensilage, depending on DM content, intensity of lactic acid formation, and ni-trate content. orchardgrass (free of nitrate) with 4 levels of DM (20, 30, 40, and 50 %) was firstly contaminated with clostridial spores about 104 / g FM. Then it was ensi-laged with following treatments: without additives, with inoculation of lactic acid bac-teria (LAB) alone or in combination with 2 % Glucose in FM (LAB+G), with nitrate addition (0.1 and 0.15 % N in DM), and with LAB plus nitrate 0.1 % N in DM. The si-lages were analysed 3, 7, 14, 28, 56, and 180 days after ensiling. Results showed that silage quality not only depends on DM content and ratio of WSC to BC, but also depends on nitrate content and extent of clostridial spores in forage. With an extremely low content of nitrate a high risk of butyric acid formation is given in silages, even if the ensiling material had a high ensilability (FC ³ 45) and a very low content of clostridial spores. The butyric acid concentration decreased with in-creasing DM content from 14 to 40 % or with increasing WSC/BC-ratio. But to get the silages free of butyric acid, a certain amount of nitrate was required. By adding clos-tridial spores in fodder the risk of butyric acid formation was increased, especially in case of material lacking in nitrate. The value of MCN depends on ensilability of the forage, as measured by DM-content and WSC/BC-ratio or by fermentability coefficient (FC) = DM+8WCS/BC, as well as depends on content of clostridial spores in the material used. The higher the FC-value and the lower the spores content is, the less nitrate is required to get silage free of butyric acid. MNC (% N in DM) for very low contaminated forage = 0.24 - 0.0035.FC MNC (% N in DM) for high contaminated forage = 0.20 - 0.021.FC High content of clostridial spores was especially found in silages containing butyric acid, which were made from forage with very low nitrate content (£ 0.02 % N in DM). But a strong relationship was not found between butyric acid and spores content. By increasing DM content the development of clostridia during ensiling was limited. A continuous decrease of spores content, in comparison with the forage before ensiled, was observed at first by increasing DM content to 50 %. By inoculation with LAB or LAB+G the lactic acid formation was strongly stimulated. A very low pH was reached 3 or 7 days after ensiling. But the butyric acid formation could be firstly prevented by increasing DM content to over 40 %. For all levels of DM, by nitrate addition the silages remained no butyric acid during the whole period of incubation. The concentration of clostridial spores decreased continuously during ensilage. This decrease was slower with increasing DM content . By combination of LAB with nitrate a reliable prevention of butyric acid formation and a fast decrease in spores concentration were reached.

Colloidal particles suspended in a liquid or gas phase often interact with a solid-liquid or solid-gas interface. In this study, experimental data through atomic force microscopy and neutron reflectometry and theoretical results of colloidal particle-surface interactions were obtained and compared. Atmospheric and aquatic environments were considered for the interactions of microbial colloidal particles and nano-sized silica particles with planar surfaces. Spores of Bacillus thuringiensis, members of the Bacillus cereus group, were examined as the microbial particles, simulating the pathogens Bacillus cereus and Bacillus anthracis which are potentially dangerous to human health. Model planar surfaces used in this study include gold which is an electrically conductive surface, mica which is a highly charged, nonconductive surface, and silica. In atmospheric systems, the interaction forces were found to be strongly affected by the relative humidity, and the total adhesion force of a particle onto a surface was modeled as the addition of the capillary, van der Waals, and electrostatic forces. Each component is influenced by the properties of the particle and surface materials, including hydrophobicity and surface roughness, as well as the humidity of the surrounding atmosphere. In aquatic systems, the interaction forces are mainly affected by the solution chemistry, including pH and ionic strength. The main components of the interaction force between a microbial colloidal particle and a planar surface were found to be the van der Waals and electrostatic forces. The results obtained in this research provide insights into the fundamental mechanisms of colloidal particle interactions with environmental surfaces in both atmospheric and aquatic systems, contributing to the understanding of the phenomena driving such interfacial processes as deposition, aggregation, and sedimentation. Thus, the results can help us describe the behavior of contaminant colloidal particles in environmental systems and subsequently devise better means for their removal from environmental surfaces.

No description available.

Viability and dispersal of ascospores of Anisogramma anomala, the cause of eastern filbert blight (EFB) on European hazelnut, from diseased branches pruned from trees were measured. In each of two years, branches bearing stromata of A. anomala were cut in mid-December and compared to branches cut near budbreak in March, when trees became susceptible to infection. The experiment was replicated three times at separated locations. At each location, 125 diseased branches (source) were piled loosely in a 1 x 1 m area. From March to June, spore traps (rain sampling-type) as well as 2-year-old potted hazelnut trees were placed next to each source, 6.4 m upwind and downwind, and 20 m downwind from each source. During seven significant major rain events over the two seasons, hazelnut seedlings (3-month-old) were placed adjacent to the spore traps. Near sources significantly higher (P. < 0.01) ascospores counts were obtained for branches cut near budbreak compared to those pruned in December in the first season; no significant difference in counts of ascospores were observed in the second season between pruning treatments. For both seasons significantly higher (P < 0.05) counts of ascospores were observed at 6.4 m downwind compared to 6.4 m upwind or 20 m downwind of a source. Ascospore viability, as assessed by staining with trypan blue, was similar for both pruning times at all distances and averaged 50%. At least one infected seedling was obtained for 5 of 7 major rain events regardless of pruning time at sources and 3 of 7 major rain events 6.4 m downwind of a source. All of the 2-year-old potted trees for both pruning treatments at the source and 6.4 m downwind became diseased and > 50% of trees at 20 m downwind became diseased in the 2010 season. Similar to ascospores counts, disease incidence was significantly higher (P < 0.01) in 2-year-old potted trees observed 6.4 m downwind compared to 6.4 m upwind or 20 m downwind in the 2010 season. Significantly higher (P < 0.01) disease incidence in 2-year-old potted trees was observed 20 m downwind compared to 6.4 m upwind in the 2010 season. Downwind disease gradients for both pruning treatments were shallow with slopes that were not significantly different than zero (p > 0.05) for the 2010 season. Based on these results, ascospores from diseased branches pruned from trees in both pruning treatments remained viable, infectious and were dispersed downwind of each treatment. / Graduation date: 2012

The data from public herbal collections were used to determine the frequency of the occurence of fertile bracken populations. Samples of bracken were collected to assess the degree of fertility and also to reveal the frequency of both diploid and triploid cytotypes using flow cytometry. Collected samples were also used for SSR analysis to characterize the population structure of both cytotypes and the propagation of triploid plants of bracken.