Orientador: José Luiz Pereira / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-21T13:27:50Z (GMT). No. of bitstreams: 1 Lopes_AlinedeSouza_M.pdf: 1681631 bytes, checksum: f77820cebe02c8058ad8b01cc2f36a6e (MD5) Previous issue date: 2012 / Resumo: O gênero Aspergillus é um grupo de fungos que possui diversas espécies produtoras de micotoxinas, distribuídas principalmente em três seções denominadas de Nigri, Flavi e Circumdati. Estudos para isolamento destas espécies estão sendo executados para se conhecer a micobiota e atuar na prevenção e redução da contaminação dos alimentos, principalmente por micotoxinas, como também são úteis nas descobertas de novas espécies. A identificação de fungos, como o gênero Aspergillus sp foi, por muito tempo,realizada através de suas características morfológicas, sendo hoje amparadas por técnicas como a Biologia Molecular, fisiologia e detecção de metabólitos específicos produzidos pelos microrganismos. Com este objetivo, este trabalho apresenta o inicio do levantamento de dados relacionado à ocorrência, caracteres morfológicos, fisiológicos,bioquímicos e moleculares, assim como a distribuição geográfica. A partir do isolamento de 10.048 cepas potencialmente toxigênicos de amostras de café, cacau, castanha do Brasil e frutas secas (tâmaras, uvas passas, figos e ameixas), matérias-primas de projetos desenvolvidos no Laboratório de Microbiologia do Instituto de Tecnologia de Alimentos (ITAL), 5.069 destes isolados foram preservadas em sílica gel, exigindo a catalogação dos dados. Neste acervo, a section Flavi predominou com o número de 2.507 culturas (32% destas cepas foram produtoras de aflatoxinas), seguida da section Nigri com 2.078 e 463 da section Circumdati que, somando, contribuiram com 11% de fungos produtores de ocratoxina A. Os Aspergillus da section Nigri apareceram em número considerável em todos os substratos, confirmando a sua predominância destes como contaminantes de alimentos. As amostras de castanha do Brasil contribuíram com o maior número de isolados, principalmente pela biodiversidade da floresta e colheita extrativista. Fungos que apresentaram estruturas diferenciadas, representantes de grupos com mesmas características, toxicidade ou espécies novas foram encaminhados para outros tipos de identificação. Duzentos e setenta e seis culturas foram identificadas por análise molecular, 435 pela extração de seus metabólitos e 87 espécies foram classificadas através da identificação polifásica. A distribuição das culturas apresentou representantes do Norte, Nordeste, Sul e Sudeste do Brasil, sendo que o Pará e Amazonas contribuíram com 2.759, como também culturas originárias de amostras de outros países como Irã,Turquia, Tunísia, EUA, México, Espanha e Argentina. A rotina de uma coleção consiste em novos isolamentos, manutenção do acervo e atualização do banco de dados, um trabalho enriquecedor para a ciência e que nunca se encerra / Abstract: The genus Aspergillus is part of a fungi group with several species that produce mycotoxins, mainly distributed in three sections named Nigri, Flavi and Circumdati. Studies to isolate these microorganism types are being made to know the mycobiota and their function in prevention and reduction of food contamination, mainly by mycotoxins and also to discover new species. The Aspergillus fungi identification was for a long time made by morphological characteristics but now it is supported by techniques such as molecular biology, physiology and detection of microorganism metabolites. With the objective this work presents the beginning of data collection related to the occurrence, morphological, physiological, biochemical and molecular, as well as the geographic distribution. From the potentially toxigenic strains isolation of 10,048 samples of coffee, cocoa, Brazil nuts and dried fruit (dates, raisins, figs and prunes) raw materials for projects developed in the Laboratory of Microbiology, Institute of Food Technology (ITAL), 5069 of these isolates were preserved in silica gel, requiring cataloging data. In this collection section Flavi predominates with 2,507 cultures (32% of these strains are aflatoxin producers) followed by section Nigri with 2,078 and 463 of section Circumdati that together, contributes 11% of ochratoxin A fungi producing. The Aspergillus section Nigri showed a considerable number of all substrates, confirming its predominance and resistance as a food contaminant. Brazil nut samples contributed with the largest number of strains due to forest biodiversity and harvest extraction. Fungi that had differentiated structures, group representatives with similar characteristics, toxicity or new species were referred to other types of identification. Two hundred and seventy six isolates were identified by molecular analysis with 435 metabolites, 88 species of Aspergillus showed the two forms, being classified by polyphasic identification. The genus Aspergillus was identified widely from countries such as Iran, Turkey, Tunisia, USA, Mexico, Spain and Argentina. In Brazil there are representatives from the North, Northeast, South and Southeast, and Para and Amazonas states that contributed to 2,759 cultures. The collection routine consists of new insolation, collection maintenance and updating of the database, which is an undending task for the enrichment of science / Mestrado / Ciência de Alimentos / Mestra em Ciência de Alimentos

Replacing fossil fuels with algae, a renewable resource, is an exciting possibility. This study evaluates the algae found in South Texas brackish water ponds used for aquaculture of fish as a possible source of biofuels. Samples of algae from these ponds were cultured at varying temperatures ranging from 15.5ºC to 36.5ºC. High levels of growth were observed at 20.5ºC and the highest lipid content was measured at 23.0ºC. Temperature was also a factor in the distribution of microalgal taxa throughout the temperature gradient. This information will be added to the growing body of research investigating similar cultures of algae for future biofuel production. / text

<p>Ispitano je prisustvo toksičnih cijanobakterija u različitim ekosistemima sa teritorije&nbsp;Republike Srbije i analizirane su negativne posledice koje mogu da prouzrokuju ovi&nbsp;mikroorganizmi. Formirana je Baza podataka cijanobakterija u Srbiji koja prilaže veliki broj bitnih&nbsp; i korisnih informacija iz preko 70 literaturnih izvora o prostiranju i učestalosti pojave cijanobakterija i njihovih toksina u periodu od 130 godina, kao i njihovih efekata na živi svet u vodenim ekosistemima, ali i &scaron;ire. Istraživana su 64 vodena ekosistema, uključujući reke, jezera, bare, kanale, ribnjake, akumulacije za navodnjavanje, akumulacije za snabdevanje vodom za piće i akumulacije sa drugom namenom, gde je najče&scaron;će cvetalo pet vrsta cijanobakterija i to:&nbsp;<em> Microcystis aeruginosa,&nbsp; Aphanizomenon flos-aquae</em>,&nbsp; <em>Planktothrix agardhii,&nbsp; Microcystis flos-aquae</em>&nbsp; i<em>&nbsp; Planktothrix rubescens</em>&nbsp; koje ujedno i &scaron;ire svoj areal rasprostranjenja na teritoriji Republike Srbije. U brojnim vodenim telima detektovani su mikrocistini i to u visokim koncentracijama.</p><p>Istraživanje vodenog ekosistema Ludo&scaron; sa teritorije Republike Srbije vr&scaron;eno je da bi se ustanovilo prisustvo i uticaj cijanobakterija i cijanotoksina na druge biljne i životinjske organizme u prirodnim uslovima. Trofički status jezera Ludo&scaron; najče&scaron;će je eutrofan, a cvetanje cijanobakterija kontinuirano jo&scaron; od 1970. godine. Tokom 2011. i 2012. godine&nbsp; vrste<em> Limnothrix&nbsp; redekei&nbsp;</em> i&nbsp;<em> Pseudanabaena&nbsp; limnetica</em>&nbsp; nađene&nbsp; u&nbsp; cvetanju. Detektovano je i prisustvo mikrocistina/nodularina i saksitoksina u biomasi i vodi, a mikrocistini su detektovani i u tkivu vodenih biljaka (trske&nbsp;<em> Phragmites communis</em>, rogoza&nbsp; <em>Typha latifolia</em>&nbsp; i ljubičastog lokvanja<em> Nymphaea elegans</em>) i ribi (<em>Carassius gibelio</em>) iz jezera Ludo&scaron;. Histolo&scaron;kim pregledom tkiva ribe&nbsp; pronađene su naizraženije promene u jetri, bubrezima i &scaron;krgama, a primećene su i na crevima.</p><p>Testiranjem biolo&scaron;kih lesnih pokorica sa teritorije Vojvodine nisu detektovani&nbsp;mikrocistini/nodularini, a nije detektovana ni toksičnost uzoraka. Pretpostavlja se da su koncentracije cijanotoksina ispod granica detekcije ili ih nema u testiranim biolo&scaron;kim lesnim pokoricama. Razvojem novih metoda i optimizacijom postojećih za detekciju cijanotoksina u biolo&scaron;kim lesnim pokoricama i drugim terestričnim ekosistemima potrebno je proveriti dobijene rezultate.</p><p>Proučavanjem svojstva 84 soja cijanobakterija iz NSCCC, koji potiču iz terestričnih i&nbsp;vodenih ekosistema sa teritorije Republike Srbije, dobijena je intracelularna toksičnost u eksponencijalnoj i stacionarnoj fazi rasta, kao i ekstracelularna toksičnost &nbsp;kod jednog soja koji potiče sa terestričnog ekosistema. Dobijeni su i pozitivni rezultati &nbsp;na prisustvo mikrocistina, nodularina ili/i saksitoksina kod 34,1% terestričnih i 55,5% vodenih sojeva koji potiču sa teritorije Republike Srbije. Rezultati ukazuju na&nbsp; potencijalnu opasnost pojave ovih&nbsp; mikroorganizama i njihovih toksičnih metabolita u različitim ekosistemima sa teritorije Republike Srbije.</p><p>Eksperimentalno je potvrđena akumulacija mikrocistina u račiću<em>&nbsp; Daphnia pulex&nbsp; </em>usled ishrane toksičnim sojem iz NSCCC, zbog čega upotreba ove vrste račića u izvođenju biolo&scaron;kih testova za testiranje prisustva cijanotoksina treba da se preispita.&nbsp; &Scaron;tavi&scaron;e, veliki broj jedinki dafnija (92,2%) koristio je istraživane sojeve iz NSCCC za ishranu, odnosno ishrana je bila moguća sa svim vodenim i sa gotovo 90% terestričnih sojeva, &scaron;to može poslužiti kao osnov za dalja istraživanja prevencije cvetanja.</p><p>S obzirom na mogućnost ishrane račića&nbsp;<em> Daphnia&nbsp; sp</em>. cijanobakterijama, kompleks&nbsp;<br />ribnjaka sa teritorije Republike Srbije kori&scaron;ćen je za istraživanje potencijalnog načina&nbsp;<br />prevencije pojave i cvetanja cijanobakterija u zavisnosti od pravovremenog uno&scaron;enja&nbsp;<br />pomenutog račića. Na osnovu koncentracije hlorofila&nbsp; a&nbsp; i trofičkog statusa,&nbsp; kvalitativne i kvantitativne analize cijanobakterija, toksičnosti vode&nbsp; i prisustva cijanotoksina mikrocistina/nodularina i saksitoksina, potvrđeno je smanjenje cvetanja cijanobakterija i drugih negativnih efekata u eksperimentalnim jezerima u odnosu na kontrolna. Ukoliko pak dođe do cvetanja i proizvodnje toksina&nbsp; ijanobakterija u vodenim ekosistemima, zbog ozbiljnost pojave i mogućih negativnih posledica po zdravlje ljudi, neohodno je uvesti postupke eliminacije ćelija&nbsp; cijanobakterija i njihovih &nbsp;toksina u praksu pri obradi&nbsp; otpadnih voda i preči&scaron;ćavanja vode iz povr&scaron;inskih akumulacija u Republici Srbiji.</p> / <p>The presence of toxic&nbsp; cyanobacteria in different ecosystems from the territory of the Republic of Serbia was analyzed as well as the negative consequences that may be causeed by these microorganisms. Serbian Cyanobacterial Data Base was formed where great number of important and useful information from over 70 literary sources regarding the distribution and frequency of cyanobacteria and their toxins over a period of 130 years, as well as their effects on wildlife in aquatic ecosystems, and beyond was presented. The study consisted of 64 aquatic ecosystems, including rivers, lakes, ponds, canals, irrigation reservoirs, reservoirs for drinking water supply and reservoirs with other purpose, where five species of cyanobacteria <em>Microcystis aeruginosa,&nbsp; Aphanizomenon flos-aquae,&nbsp; Planktothrix agardhii,&nbsp; Microcystis flosaquae&nbsp;</em> and&nbsp; <em>Planktothrix rubescens</em>&nbsp; frequently bloomed, and also expanded their area of distribution on the territory of the Republic of Serbia. In many water bodies microcystins were detected in high concentrations.</p><p>Research into aquatic ecosystem Ludo&scaron;, located on the territory of the Republic of Serbia, was performed in order to determine the presence and effect of cyanobacteria and cyanotoxins on other plant and animal organisms in natural conditions. Trophic status of&nbsp; the lake Ludo&scaron; was usually eutrophic, and cyanobacteria bloom is constant since 1970. During 2011 and 2012, the species <em>Limnothrix&nbsp; redekei</em>&nbsp; and&nbsp; <em>Pseudanabaena&nbsp; limnetica&nbsp;</em> were found in blooming. Presence of microcystins/nodularin and saxitoxin was detected in&nbsp; biomass and water, and microcystins were detected in tissues of aquatic plants (reed <em>Phragmites communis,</em> cattail&nbsp;<em>Typha latifolia&nbsp;</em>and royalblue waterlily&nbsp; <em>Nymphaea elegans</em>) and fish (<em>Carassius gibelio</em>) from the lake Ludo&scaron;. Histological examination of tissue showed most prominent changes in liver, kidney and gills, and alterations were also observed in the intestine.</p><p>Testing of the biological loess crust from Vojvodina showed no presence of microcystins/nodularin and toxicity of samples was not detected as well. It is assumed that cyanotoxin concentrations were below detection limit or are absent from the tested biological loess crusts. The development of new methods and optimization of existing ones for detection of cyanotoxins in biological loess crustsand&nbsp; other terrestrial ecosystems is necessary in order to revise obtained results.</p><p>Research of the properties from 84 strains of cyanobacteria from NSCCC originating&nbsp;from terrestrial and aquatic ecosystems from the territory of the Republic of &nbsp;Serbia, resulte d inintracellular toxicity in exponential and stationary growth phase, as &nbsp;well as extracellular toxicity of a strain originating from terrestrial ecosystems. The obtained results were positive for the presence of microcystins, nodularin and/or saxitoxin&nbsp; for 34.1% terrestrial and 55.5% aquatic strains originating from the territory of the Republic of Serbia. These results demonstrate the potential risk of occurrence of these microorganisms and their metabolites in different ecosystems from the territory of the Republic of Serbia.</p><p>Accumulation of microcystins in shrimp <em>Daphnia pulex</em>&nbsp;after feeding with toxic strain&nbsp;from NSCCC was experimentally confirmed, indicating that the use of this species of shrimp in biological tests which determine the presence of cyanotoxins needs to be revisited. Moreover, a large number of&nbsp;<em>Daphnia&nbsp; individuals </em>(92.2%) used the investigated strains of NSCCC for food, and the feeding was possible with all &nbsp;the water strains and nearly 90% of terrestrial strains, which can serve as a basis for further research of bloom prevention.</p><p>With regard to the possibility shrimp <em>Daphnia&nbsp;</em>sp. feeding with cyanobacteria, a&nbsp;complex of ponds from the territory of the Republic of Serbia was used to explore &nbsp;potential ways of prevention the occurrence and blooming of cyanobacteria, depending on the timely introduction of the aforementioned shrimp. Based on the concentrations of chlorophyll&nbsp; a&nbsp; and trophic status, qualitative and quantitative analysis of cyanobacteria, the toxicity of water and the presence of&nbsp; cyanotoxins microcystins/nodularin and saxitoxin, reduction in blooming cyanobacteria and other negative effects in the experimental lakes when compared to the control lakes was confirmed. If the blooming regardless occurres as well as production of cyanobacterial &nbsp;toxins in aquatic ecosystems, due to the seriousness of the phenomenon and the possible negative consequences for human health, it would be necessary to introduce &nbsp;procedures for the elimination of cells of cyanobacteria and their toxins into practice in waste water treatment and purification of water from surface reservoirs in the Republic of Serbia.</p>

Ziel der vorliegenden Arbeit war die Untersuchung von Mikroalgen-Kulturstämmen unterhalb des Artniveaus. Für Service-Kulturensammlungen sind diese Untersuchungen wichtig, um die genetische Diversität der vorhandenen bzw. neu aufzunehmender Kulturen zu erfassen, Stämme eindeutig zu identifizieren oder auch Kontaminationen auszuschließen. Zu diesem Zweck wurden häufig in der Forschung genutzte Stämme von Mikroalgen mithilfe von genetischen Fingerabdrücken, die mit der AFLP (amplified fragment length polymorphism) Technik generiert wurden, untersucht. Mittels AFLP was es möglich, verschiedene Stämme derselben Art (Chlorella vulgaris) zu unterscheiden und für jedes Isolat einen charakteristischen genetischen Fingerabdruck zu erhalten. Da diese genomischen Variationen innerhalb derselben Art auch mit Unterschieden in physiologischen/biochemischen Eigenschaften korreliert sein können, ist es wichtig genau festzuhalten, welches Isolat für einen Versuch oder eine biotechnologische Anwendung verwendet wurde. Mit der AFLP-Technik war es ferner möglich, phänotypisch unterschiedliche Mutanten von Parachlorella kessleri und den entsprechenden Wildtyp zu unterscheiden. Normalerweise ist es mit AFLP nicht möglich, Algenkulturen zu untersuchen, die mit anderen Organismen kontaminiert sind. Das Problem hierbei ist, dass bei den generierten Fragmenten nicht unterschieden werden kann, ob sie von der Alge stammen oder von der Kontamination. Es konnte hier jedoch gezeigt werden, dass kontaminierte Kulturen trotzdem mit AFLP untersucht werden können. Hierzu war es nötig, Kulturen mit unterschiedlichem Verhältnis von Algen- und Bakterienzellen zu untersuchen. Außerdem konnte gezeigt werden, dass Infektionen von Algenkulturen mit Viren, die meistens schwer nachzuweisen sind, mittels AFLP festgestellt werden können. Kryokonservierung ist mittlerweile die Methode der Wahl zur Langzeitaufbewahrung von Mikroalgen. Es wird davon ausgegangen, dass diese Aufbewahrungsmethode genetisch stabile Kulturen über Jahrzehnte hinweg garantieren kann. Jedoch wurde bis heute die genetische Stabilität von kryokonservierten Algen noch nicht überprüft und deshalb in der vorliegenden Arbeit mithilfe der AFLP-Technik untersucht.