Se desarrollaron películas biodegradables activas a base de PHBV, combinadas con otros biopolímeros (PLA y almidón) y diferentes compuestos antimicrobianos (aceites esenciales (AE)), las cuales se caracterizaron en cuanto a sus propiedades funcionales y estructurales a fin de obtener materiales que satisfagan mejor los requisitos de envasado de alimentos. La plastificación del PHBV se llevó a cabo mediante el uso de diferentes compuestos con el objetivo de mejorar el rendimiento mecánico de los films. Asimismo, se incorporaron diferentes activos (AE de orégano y clavo, así como sus respectivos compuestos mayoritarios, carvacrol (CA) y eugenol (EU)) en películas bicapa de PHBV pulverizando los activos entre dos monocapas obtenidas por termocompresión. También se analizó la potencial sinergia entre diferentes compuestos de AE, así como sus aplicaciones a diferentes alimentos cuando se incorporan en películas bicapa de PHBV. Se desarrollaron películas antimicrobianas multicapa donde se combinaron láminas polares (almidón), apolares (poliéster) y CA, ya sea pulverizado entre ambas capas o incorporado en la solución filmogénica de poliésteres, a fin de optimizar la funcionalidad del material. El proceso de valorización de la cascarilla de arroz, basado en extracción con agua subcrítica, permitió obtener xilanos bioactivos y fracciones celulósicas con potenciales aplicaciones en el envasado de alimentos. Pese a que la adición de polietilenglicol (PEG) de diferente peso molecular o ácido láurico disminuyó significativamente la rigidez y la resistencia de las películas de PHBV, solo el PEG1000 dio lugar a películas más extensibles. No obstante, estrategias adicionales fueron necesarias a fin de adaptar las propiedades mecánicas de los films de PHBV a ciertos requisitos de envasado. La pulverización de activos en la interfaz de ambas monocapas de PHBV generó películas antimicrobianas con propiedades físicas adecuadas. Además, la liberación de los activos desde las películas permitió controlar el crecimiento de Escherichia coli y Listeria innocua en condiciones in vitro. Ambos activos, CA y EU, se liberaron de manera efectiva en diferentes simulantes alimentarios. En este sentido, la tasa de liberación mejoró cuando disminuyó la polaridad de los simulantes. El efecto sinérgico más notable para los compuestos de AE se observó para las mezclas CA/cinnamaldehído para ambas bacterias, pero usando diferentes proporciones de compuestos. De esta forma, los resultados obtenidos permitieron la optimización de la dosis de activos utilizados para la aplicación de alimentos, minimizando así su impacto sensorial. Las películas de PHBV con compuestos activos de AE fueron altamente efectivas contra L. innocua y E. coli in vitro, pero mucho menos activas en alimentos. Asimismo, no se observó ninguna correlación entre la cantidad de activo migrada al alimento y el efecto antibacteriano en las diferentes matrices, lo que refleja que existen numerosos factores composicionales que afectan a la disponibilidad de los activos a la hora de ejercer su acción antibacteriana sobre un alimento determinado. La formulación 75:25 PLA-PHBV con PEG1000 exhibió las mejores propiedades físicas y, por lo tanto, se utilizó como soporte del CA mediante casting, así como para desarrollar bicapas con almidón. La incorporación de CA pulverizarlo entre las láminas de poliéster y almidón no fue eficaz a la hora de retener el compuesto activo en las bicapas. Sin embargo, la incorporación de CA en películas de poliésteres mediante casting dio lugar a una retención de CA prácticamente total, lo que condujo a una notable actividad antimicrobiana. Además, estas bicapas exhibieron propiedades físicas altamente mejoradas con respecto a la monocapa de almidón. La valorización de la cáscara de arroz mediante extracción con agua subcrítica, permitió obtener hemicelulosas mejor conservadas, con actividad antioxi / Biodegradable active films based on PHBV, combined with other biopolymers (PLA and starch) and different antimicrobial compounds (essential oils (EO)), were developed and characterized as to their functional and structural properties to obtain materials that better meet food packaging requirements. Plasticization of PHBV was analysed by using different compounds to enhance the PHBV mechanical performance. Likewise, different active compounds (oregano and clove EO, as well as their respective main compounds, carvacrol (CA) and eugenol (EU)) were incorporated into PHBV bilayer films by spraying the actives between two thermo-compressed monolayers. The potential synergy between different EO compounds and their applications to different food when incorporated into PHBV films was also analysed. Multilayer antimicrobial films combining polar (starch) and non-polar (polyester) sheets, incorporating CA, either by spraying it between both layers or incorporating it in the polyester casting solution, were developed to optimise the material functionality. Bioactive xylans and cellulosic fractions from rice husk, which are useful for food packaging applications, were obtained by using an eco-friendlier valorization process based on subcritical water extraction. Although the addition of polyethylene glycol (PEG) of different molecular weight and lauric acid significantly decreased the stiffness and the resistance to break of PHBV films, only PEG1000 yielded more extensible films. Nonetheless, additional strategies would be required to adapt PHBV mechanical properties to certain packaging requirements. Spraying actives at the interface of both PHBV monolayers produced antimicrobial films with appropriate physical properties. The release of the actives from the films was adequate to control the growth of E. coli and L. innocua in vitro. Both actives, CA and EU were effectively released into different food simulants. The release rate was enhanced when the polarity of the simulants decreased, but it fell markedly in fatty systems. The most remarkable synergistic effect for the EO compounds was observed for CA/cinnamaldehyde blends for both bacteria but using different compound ratios. Thus, the results allowed for the optimization of the dose of actives used for food application, thus minimizing their sensory impact. PHBV films with active EO compounds were highly effective against L. innocua and E. coli in the in vitro tests, but they were much less effective in foods. Likewise, no correlation between the amount of active that migrated to the food and the antibacterial effect was observed, which reflected that many compositional factors affect the availability of the antimicrobials to exert their action on a specific food. The 75:25 PLA-PHBV formulation with PEG1000 exhibited the best properties in terms of physical properties and, thus it was used to be the carrier of CA by casting and to develop bilayers with starch. Incorporating CA by spraying it between the polyester and starch sheets was not effective at retaining this active in the bilayers. However, the incorporation of CA into casted polyester films was highly effective at providing practically total CA retention, which led to a notable antimicrobial activity. Moreover, these bilayers exhibited highly improved tensile and water vapour barrier capacity with respect to the starch monolayer. The rice husk valorization, based on subcritical water extraction, allowed for obtaining better preserved hemicelluloses, with antioxidant and antibacterial activity, useful as additives for food or food packaging applications, and cellulosic reinforcing agents to develop biocomposites with enhanced mechanical performance. / Es van desenvolupar pel·lícules biodegradables actives a base de PHBV, combinades amb altres biopolímers (PLA i midó) i diferents compostos antimicrobians (olis essencials (OE)), les quals es van caracteritzar quant a les seues propietats funcionals i estructurals a fi d'obtindre materials que complisquen millor els requisits d'envasament d'aliments. La plastificació del PHBV es va dur a terme per mitjà de diferents compostos amb l'objectiu de millorar el rendiment mecànic dels films. Així mateix, es van incorporar diferents actius (OE d'orenga i clau, així com els seus respectius compostos majoritaris, carvacrol (CA) i eugenol (EU)) en pel·lícules bicapa de PHBV polvoritzant els actius entre dos monocapas, obtingudes per termocompressió. També es va analitzar la potencial sinergia entre diferents compostos d'OE, així com les seues aplicacions a diferents aliments quan s'incorporen en pel·lícules bicapa de PHBV. Es van desenvolupar pel·lícules antimicrobianes multicapa on es combinaren làmines polars (midó) i apolars (polièsters) i carvacrol, ja siga polvoritzat entre ambdós capes o incorporat en la solució filmogénica de polièsters, a fi d'optimitzar la funcionalitat del material. El procés de valoració de la corfeta d'arròs, basat en extracció amb aigua subcrítica, va permetre obtindre xilans bioactius i fraccions cel·lulòsiques amb potencials aplicacions en l'envasament d'aliments. A pesar que l'addició de polietilenglicol (PEG) de diferent pes molecular o àcid làuric va disminuir significativament la rigidesa i la resistència de les pel·lícules de PHBV, només PEG1000 va donar lloc a pel·lícules més extensibles. No obstant això, estratègies addicionals varen ser necessàries a fi d'adaptar les propietats mecàniques dels films de PHBV a certs requisits d'envasament. La polvorització d'actius a la interfase d'ambdós monocapes de PHBV va generar pel·lícules antimicrobianes amb propietats físiques adequades. L'alliberament dels actius des de les pel·lícules va permetre controlar el creixement d'Escherichia coli i Listeria innocua en assajos in vitro. Ambdós actius, CA i EU, es van alliberar de manera efectiva en els diferents simulantes alimentaris. La taxa d'alliberament va millorar quan va disminuir la polaritat dels sistemes aquosos. L'efecte sinèrgic més notable per als compostos d'OE es va observar per a les mescles de CA/cinnamaldehído per a ambdós bacteris, però utilitzant diferents proporcions. D'esta manera, els resultats van permetre l'optimització de la dosi d'actius utilitzats per a l'aplicació en aliments, minimitzant així el seu impacte sensorial. Les pel·lícules de PHBV amb actius d'OE van ser altament efectives front L. innocua i E. coli en les proves in vitro, però van ser molt menys efectives en aliments. Així mateix, no es va observar cap correlació entre la quantitat d'actiu migrada a l'aliment i l'efecte antibacterià en les diferents matrius, la qual cosa reflectix que hi ha molts factors composicionals que afecten la efectivitat del dels compostos actius per a exercir la seua acció sobre un aliment en concret. La formulació 75:25 PLA-PHBV amb PEG1000 va exhibir les millors propietats físiques i, per tant, es va utilitzar com a suport del CA per mitjà de càsting, així com per a desenvolupar bicapes amb midó. La incorporació de CA polvoritzat entre les làmines de polièsters i midó no va ser eficaç per a retindre el compost actiu en la bicapa. No obstan, la incorporació de CA en pel·lícules de polièster per càsting va permetre una retenció de CA pràcticament total, la que va conduir a una notable activitat antimicrobiana. A més, estes bicapas van exhibir una capacitat de barrera al vapor d'aigua i a la tracció altament millorades respecte a la monocapa de midó. La valoració de la corfa d'arròs, basada en l'extracció amb aigua subcrítica, va permetre obtindre fraccions hemicelulósiques millor conservades, amb activ / Requena Peris, R. (2018). Multilayer biodegradable active films based on PHBV for food packaging [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/110079 / TESIS

Tesis por compendio / [ES] El uso masivo de plásticos sintéticos y su impacto medioambiental obliga a buscar alternativas biodegradables para el envasado de los alimentos,etapa necesaria para su adecuada conservación.Así mismo,la necesidad de incrementar la vida útil de los alimentos ha despertado gran interés en el desarrollo de materiales activos(antimicrobianos y antioxidantes)que mantengan su calidad y seguridad por más tiempo,mediante el uso de compuestos de origen natural,seguros para el consumidor.En este sentido,el desarrollo de materiales biodegradables activos para el envasado de alimentos constituye hoy en día un reto importante para la industria alimentaria.En la presente Tesis Doctoral,se ha estudiado la encapsulación de carvacrol mediante el electroestirado o extensión y secado de diferentes disoluciones poliméricas con carvacrol.Se han utilizado polímeros biodegradables portadores de diferente polaridad(almidón termoplástico:TPS,polivinil-alcohol:PVA, policaprolactona:PCL o ácido poliláctico:PLA)disueltos en el solvente adecuado,con el fin de obtener capas activas.Estas capas se han combinado con otras de polímeros con propiedades complementarias,para obtener laminados activos adecuados para el envasado de alimentos.Los laminados combinaron polímeros polares(TPS o PVA)y poliésteres no polares(PCL o PLA)incorporando el carvacrol en una de las capas.Se evaluó la cinética de liberación del activo,así como la acción antimicrobiana de los materiales obtenidos.Los laminados se caracterizaron en su funcionalidad como material de envase(prop. de barrera,mecánicas u ópticas)así como en su estructura y comportamiento térmico.Los estudios de encapsulación revelaron un mayor potencial encapsulante del carvacrol para los polímeros no polares(PCL;PLA),aunque el PVA mostró también una buena afinidad con el compuesto activo.La matriz de PVA mostró una mayor retención de carvacrol mediante electroestirado de sus disoluciones acuosas que por extensión y secado,sin necesidad de adición de tensoactivos como el Tween85.Para la encapsulación en PLA,se usaron mezclas binarias de solventes aptos para contacto con los alimentos(acetato de etilo y DMSO).En este caso,se obtuvo una mayor eficiencia encapsulante del PLA en los materiales obtenidos por extensión y secado que en los electroestirados.La cinética de liberación del carvacrol de las fibras de PCL explicó el mayor efecto antibacteriano contra E.coli,y el escaso efecto antilisteria.La velocidad de liberación del activo aumentó cuando disminuyó la polaridad de los simulantes alimentarios, mostrando una liberación completa en los sistemas apolares,pero solo hasta 75% en los sistemas acuosos,que requerirían una mayor proporción del activo en el envase para potenciar su efectividad.La combinación de láminas de TPS con fibras de PCL cargadas con carvacrol dio lugar a materiales con una permeabilidad al vapor de agua mejorada,en comparación con los films de almidón,sin efectos relevantes sobre las otras propiedades funcionales estudiadas.Cuando los laminados se probaron in vitro contra cepas G(+) y G(-) mostraron un efecto antibacteriano similar al de las fibras de PCL con carvacrol,pero retrasado en el tiempo.Los estudios de desintegración-biodegradación de los laminados almidón-PCL revelaron que las películas con carvacrol afectaron la actividad del inóculo del compost,disminuyendo ligeramente la biodegradabilidad de las películas,pero alcanzando valores de desintegración similares(75-80%)a las muestras libres de carvacrol.Se obtuvieron también laminados de PLA y PVA mediante la extensión y secado de disoluciones acuosas de PVA con carvacrol.La superficie del PLA fue sometida a aminolización a fin de mejorar la extensibilidad de las disoluciones acuosas.A pesar del incremento de la componente polar de la energía superficial del PLA y su mejorada humectabilidad con las soluciones de PVA,estas bicapas no mostraron una mejora significativa en las propied / [CA] L'ús massiu de plàstics sintètics i el seu impacte mediambiental obliga a buscar alternatives biodegradables per a l'envasament dels aliments necessari per a la seua conservació.Així mateix,la necessitat d'incrementar la vida útil dels aliments ha despertat gran interés en el desenvolupament de materials actius(antimicrobians i antioxidants)que mantinguen la seua qualitat i seguretat per més temps,per mitjà de l'ús de compostos d'origen natural,segurs per al consumidor.En este sentit,el desenvolupament de materials biodegradables actius per a l'envasament d'aliments constituïx un repte important per a la indústria alimentària.En la present Tesi Doctoral,s'ha estudiat l'encapsulació de carvacrol per mitjà de l'electroestirat o extensió i assecat de diferents dissolucions polimèriques amb carvacrol.S'han utilitzat polímers biodegradables portadors de diferent polaritat(midó termoplàstic:TPS, polivinil-alcohol:PVA, policaprolactona:PCL o àcid poliláctic:PLA)dissolts en el solvent adequat,a fi d'obtindre capes actives.Estes s'han combinat amb altres de polímers amb propietats complementàries,per a obtindre laminats actius adequats per a l'envasament d'aliments.Els laminats van combinar polímers polars(TPS o PVA)i poliésters no polars(PCL o PLA)incorporant el carvacrol en una de les capes.Es va avaluar la cinètica d'alliberament de l'actiu,així com l'acció antimicrobiana dels materials obtinguts.Els laminats es van caracteritzar en la seua funcionalitat com a material d'envàs(propietats de barrera, mecàniques o òptiques),així com en la seua estructura i comportament tèrmic.Els estudis d'encapsulació van revelar un major potencial encapsulant del carvacrol per als polímers no polars(PCL i PLA),encara que el PVA va mostrar també una bona afinitat amb el compost actiu.La matriu de PVA va mostrar una major retenció de carvacrol per mitjà d'electroestirat de les seues dissolucions aquoses que per extensió i assecat,sense necessitat d'addició de tensioactius com el Tween 85.Per a l'encapsulació en PLA,es van usar mescles binàries de solvents aptes per a contacte amb els aliments(acetat d'etil i DMSO).Es va obtindre una major eficiència encapsulant del PLA en els materials obtinguts per extensió i assecat que en els electroestirats.La cinètica d'alliberament del carvacrol de les fibres de PCL va explicar el major efecte antibacterià contra Escherichia coli,i l'escàs efecte antilisteria.La velocitat d'alliberament de l'actiu va augmentar quan va disminuir la polaritat dels simulants alimentaris,mostrant un alliberament complet en els sistemes no polars, però només fins a un 75% en els sistemes aquosos,que requeririen una major proporció de l'actiu en l'envàs per a potenciar la seua efectivitat.La combinació de làmines de TPS amb fibres de PCL carregades amb carvacrol va donar lloc a materials amb una permeabilitat al vapor d'aigua millorada,en comparació amb els films de midó, sense efectes rellevants sobre les altres propietats funcionals.Quan els laminats es van provar in vitro contra ceps Gram(+) i Gram(-) van mostrar un efecte antibacterià semblant al de les fibres de PCL amb carvacrol,però retardat en el temps.Els estudis de desintegració-biodegradació dels laminats midó-PCL van revelar que les pel·lícules amb carvacrol van afectar l'activitat de l'inocule del compost,disminuint lleugerament la biodegradabilitat,però aconseguint valors de desintegració semblants(75-80%)a les mostres lliures de carvacrol.Es van obtindre també laminats de PLA i PVA per mitjà de l'extensió i assecat de dissolucions aquoses de PVA amb carvacrol.La superfície del PLA va ser sotmesa a aminolizatció a fi de millorar l'extensibilitat de les dissolucions aquoses.A pesar de l'increment de la component polar de l'energia superficial del PLA i la seua millorada mullabilitat amb les solucions de PVA,estes bicapes no van mostrar una millora significativa en les propietats mecàniques i de barrera / [EN] The massive use of synthetic plastics and their environmental impact makes necessary the search for biodegradable alternatives for food packaging. Likewise, the need to increase the shelf life of food has aroused great interest in the development of active materials (antimicrobial and antioxidant) that maintain food quality and safety for longer periods of time through the use of compounds of natural origin, safe for the consumer. In this sense, the development of active biodegradable materials for food packaging is both a major imperative and challenge for the food industry today. In the present Doctoral Thesis, the encapsulation of carvacrol has been studied by means of the electrospinning or casting of different polymeric solutions with carvacrol. Biodegradable polymers with different polarities (thermoplastic starch: TPS, poly(vinyl-alcohol): PVA, poly-(¿-caprolactone): PCL or poly(lactic acid): PLA) dissolved in the appropriate solvent have been used to obtain active layers. These have been combined with other polymers with complementary properties, to obtain active laminates suitable for food packaging. The laminates combined polar polymers (TPS or PVA) and non-polar polyesters (PCL or PLA) incorporating carvacrol in one of the layers. The release kinetics of the active ingredient was evaluated, as well as the antimicrobial action of the materials obtained. The laminates were characterized in their functionality as a packaging material (barrier, mechanical or optical properties), as well as in their structure and thermal behaviour. Encapsulation studies revealed a higher encapsulating potential of carvacrol for non-polar polymers (PCL and PLA), although PVA also showed a good affinity with the active compound. The PVA matrix showed a higher retention of carvacrol by electrospinning of its aqueous solutions than by casting, without the need for addition of surfactants such as Tween 85. For the encapsulation in PLA, binary mixtures of solvents suitable for food contact (ethyl acetate and DMSO) were used. A higher encapsulation efficiency of PLA was obtained in the materials produced by casting than by electrospinning. The carvacrol release kinetics of PCL fibres explained the higher antibacterial effect against Escherichia coli and the lower antilisterial effect. The release ratio of the active ingredient increased when the polarity of the food simulants decreased, showing a complete release in non-polar systems and only up to 75% in aqueous systems that would require a higher proportion of the active ingredient in the packaging material to enhance its effectiveness. The combination of TPS films with carvacrol loaded PCL fibres resulted in materials with improved water vapour permeabilities, compared to starch films, with no relevant effects on the other functional properties. When the laminates were tested in vitro against Gram (+) and Gram (-) strains, they showed a similar antibacterial effect to that of PCL fibres with carvacrol, but delayed in time. Disintegration-biodegradation studies of PCL-starch laminates revealed that carvacrol films affected the activity of the compost inoculum, slightly decreasing the biodegradability of the laminates, but reaching similar disintegration values (75-80%) to the carvacrol-free samples. PLA and PVA laminates were also obtained by casting aqueous PVA solutions with carvacrol. The surface of PLA was submitted to aminolization in order to improve the extensibility of the aqueous solutions. Despite the increase in the polar component of the PLA surface energy and its improved wettability with PVA solutions, these bilayers did not show significant improvement in mechanical and barrier properties over the PLA monolayers. / The authors would like to thank the Ministerio de Economia y Competitividad of Spain, for funding this study as part of projects AGL2013-42989-R and AGL2016-76699-R and predoctoral research grant # BES-2014-068100. / Tampau, A. (2020). Carvacrol encapsulation by electrospinning or solvent casting to obtain biodegradable multilayer active films for food packaging applications [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/140313 / TESIS / Compendio

Es wurden zwei Feldstudien durchgeführt, um zu ermitteln, ob der Verlauf von Eimeria spp.-Infektionen bei Zicklein bei natürlicher Exposition durch die Beimischung von Zusätzen mit ätherischen Ölen (EIMERICOX®= EO1, NEXT Enhance® 200 = EO2) in den Milchaustauscher (MAT) oder das Einbringen eines kommerziellen Pulvers (Stalosan® F = STA) in die Einstreu beeinflusst werden kann. Als Kriterien zur Bewertung der Wirksamkeit dienten die Ausscheidung von Eimeria-Oozysten, die Kotkonsistenz und die klinische Symptomatik sowie die Gewichtsentwicklung. Des Weiteren wurde das Spektrum der caprinen Eimeria-Spezies in der untersuchten Population erfasst. In Studie 1 wurden in einem niederländischen Aufzuchtbetrieb 45 Zicklein verschiedener Herkunft zu je 15 Studientieren auf die Gruppen A1 (Kontrolle), B1 (EO1) und C1 (STA) aufgeteilt. In Studie 2 wurden in einem deutschen Milchziegenbetrieb 45 Zicklein aus betriebseigener Nachzucht in analoger Weise auf die Gruppen A2 (Kontrolle), B2 (EO1) und C2 (EO2) randomisiert. Die Zusätze auf Basis ätherischer Öle wurden während des Studienzeitraums kontinuierlich dem MAT beigemischt (EO1: 4 g/kg MAT; EO2: 125 g/t MAT) und über die MAT-Tränke ad libitum angeboten. Das Einstreupulver wurde zweimal wöchentlich in die Einstreu eingebracht. Insgesamt wurden in Studie 1 (53 Studientage) 484 Kotproben und in Studie 2 (56 Studientage) 847 Kotproben gesammelt, hinsichtlich ihrer Konsistenz bewertet und parasitologisch sowie teilweise differentialdiagnostisch untersucht. Die koproskopischen Untersuchungen umfassten die quantitative Bestimmung der ausgeschiedenen Oozysten pro Gramm Kot (OpG) und die Speziesdifferenzierung. Ferner wurden die Zicklein in Studie 1 am Studientag (ST) 0 und ST 35 und in Studie 2 am ST 0, ST 14, ST 35 und ST 56 gewogen und es erfolgte eine tägliche Kontrolle des Allgemeinbefindens. In beiden Studien wurden natürliche Infektionen der Zicklein mit Eimeria spp. beobachtet. In Studie 1 konnte bei 20 (44,4 %) und in Studie 2 bei 37 (82,2 %) der 45 Studientiere eine Oozystenausscheidung nachgewiesen werden. Die Befallsraten in den Kontrollgruppen (A1 und A2) lagen bei 73,3%in Studie 1 und 93,3%in Studie 2, was einen hohen bis sehr hohen Infektionsdruck anzeigt. Klinisch manifeste Eimeriosen mit Diarrhoe konnten während beider Studien nicht beobachtet werden. In Studie 2 wurde zwar bei 8,1 bis 8,5% der Kotproben in den drei Gruppen Durchfall festgestellt, dieser stand aber in keinem Zusammenhang mit einer Oozystenausscheidung von Eimeria spp. im Allgemeinen oder der pathogenen Spezies E. ninakohlyakimovae im Besonderen. Die Infektionen verliefen also ausnahmslos subklinisch, dennoch ist angesichts der hohen Befallsraten und der Präsenz pathogener Arten das Risiko eines Ausbruchs klinischer Kokzidiosen gegeben. Insgesamt handelte es sich im Falle von Oozystenausscheidung mehrheitlich um Polyinfektionen (Studie 1: 90,0 %; Studie 2: 94,4 %). In beiden Beständen wurde die Präsenz folgender neun Eimeria (E.) spp. nachgewiesen: E. alijevi, E. ninakohlyakimovae, E. hirci, E. arloingi, E. jolchijevi, E. christenseni, E. caprina, E. caprovina. Die höchste Prävalenz in den positiven Proben zeigten (in beiden Studien) E. arlongi (Studie 1: 52,5 %; Studie 2: 78,1 %) und E. ninakohlyakimovae (Studie 1: 55,9 %; Studie 2: 47,8 %). Am seltensten traten E. caprovina (5,1 %) und E. apsheronica (3,4 %) in Studie 1 sowie E. caprina (3,9 %) und E. caprovina (1,7 %) in Studie 2 auf. Die Intensität der Oozystenausscheidung bewegte sich bei 78,0% der positiven Proben in Studie 1 und 88,9% der positiven Proben in Studie 2 im niedrigen bis moderaten Bereich von 50 bis 10 000 OpG. Eine Oozystenausscheidung von mehr als 100 000 OpG (sehr hoher Bereich) trat in Studie 2 bei 1,1% der positiven Proben auf. Die Maxima in Einzelkotproben lagen in Studie 1 bei 67 000 OpG und in Studie 2 bei 157 000 OpG. Anlässlich der Prüfung möglicher Auswirkungen von STA fiel auf, dass gegen Ende des Studienzeitraums in der Gruppe C1 weniger (Re-) Infektionen im Vergleich zur Kontrolle auftraten. Auch die OpG-Werte erwiesen sich bei Auswertung der kumulierten Daten für den gesamten Studienzeitraum und den Zeitraum ab ST 47 sowie am ST 51 in Gruppe C1 als signifikant geringer als in Gruppe A1. Hinsichtlich der Untersuchung von Effekten durch den Einsatz von EO1 und EO2 wurden bei der Analyse von Befallsextensität und Ausscheidungsintensität in Studie 1 in Gruppe B1 niedrigere Befallsraten festgestellt und bei der Auswertung der über den Studienzeitraum kumulierten Daten und gegen Ende der Studie am ST 47, ST 49, ST 51 und ST 52 signifikant weniger positive Kotproben und geringere OpG- Werte als in Gruppe A1 ermittelt. Bei den Befallsraten und der kumulierten Anzahl positiver Proben konnten solche Effekte in Studie 2 nicht reproduziert werden. Gleichwohl zeigte sich am ST 35 und ST 37 in Gruppe B2 eine signifikant niedrigere Eimeria-Prävalenz als in Gruppe A2 und die Intensität der Oozystenausscheidung war in Gruppe B2 am ST 37 und in Gruppe C2 für den Zeitraum ab ST 26 sowie am ST 35, ST 37 und ST 42 signifikant geringer als in Gruppe A2. Bei den mittleren Zunahmen der Körpergewichte fielen stets leichte Gruppenunterschiede zu Gunsten der behandelten Gruppen auf (Gruppe A1: 4,33 kg, Gruppe B1: 5,58 kg, Gruppe C1: 4,55 kg und Gruppe A2: 9,70 kg, Gruppe B2: 10,39 kg, Gruppe C2: 10,74 kg) sie waren jedoch nicht statistisch signifikant. Der subjektive Eindruck einer besseren körperlichen Kondition und Gesamtentwicklung der EO1-behandelten Zicklein wurde dagegen in beiden Studien für einige Parameter und Studientage als signifikant bestätigt. Am Ende der Studien fielen zwölf der 14 Studientiere in Gruppe B1 (Studie 1) und zehn der 14 Studientiere in Gruppe B2 (Studie 2) durch einen exzellenten Entwicklungszustand auf, während dies in den Kontrollgruppen auf drei von zwölf Zicklein in Gruppe A1 (Studie 1) und fünf von 15 Zicklein in Gruppe A2 (Studie 2) zutraf. Der Einsatz von Stalosan® F in Studie 1 schien sich günstig auf den Verlauf der Kokzidiose auszuwirken. Der Eindruck einer kokzidioziden Wirkung sollte jedoch mittels weiterer Studien überprüft werden. Durch die Anwendung der Zusätze auf Basis ätherischer Öle, EIMERICOX® und NEXT Enhance® 200, konnte die Oozystenausscheidung unter subklinischen Bedingungen zwar begrenzt beeinflusst werden, bei einer massiven Exposition ist allerdings kaum davon auszugehen, dass eine Behandlung mit diesen untersuchten ätherischen Ölen eine ausreichend protektive Wirkung hätte, um Eimeriosen effektiv kontrollieren zu können. Bei einer geringen bis moderaten Expositionssituation könnten die geprüften alternativen Maßnahmen ergänzend zu weiteren prophylaktischen Maßnahmen (insbesondere im Haltungs- und Hygienemanagement) dem Aufbau eines kritischen Infektionsdrucks entgegen wirken und somit ein opportunes Anwendungsgebiet finden.

La contamination des surfaces par des bactéries et l’émergence de souches résistantes aux antimicrobiens sont des problèmes très préoccupants dans différents domaines tel que les domaines hospitalier et alimentaire. Cette contamination commence par l’adhésion de bactéries pathogènes sur une surface jusqu’à la formation de biofilms. Ces derniers contribuent à l’émergence de résistances de certaines souches bactériennes aux traitements conventionnels. Pour répondre à ces problèmes de contamination des surfaces, ces travaux de thèse portent sur le développement de nouveaux matériaux antibactériens à base de fibres de pâte à papier. Nous nous sommes intéressés, dans une première partie, à l’élaboration d’un papier antibactérien par le greffage, via un lien triazine, de deux composés d’huiles essentielles, le thymol et le carvacrol, connus pour leurs activités antibactériennes. L’évaluation microbiologique des matériaux élaborés, sur les deux souches bactériennes testées, E.coli et S.aureus, a montré un effet bactériostatique. Ces matériaux bloquent donc la croissance bactérienne empêchant ainsi la formation des biofilms. Une synergie entre le thymol et le carvacrol lorsqu’ils sont greffés sur les fibres de pâte à papier a également été montré. Dans une deuxième partie, notre étude s’est focalisée sur l’élaboration d’un papier antibactérien qui n’acquière son activité qu’après greffage et formation du motif actif « aryl-1,2,3-triazole ». Le greffage est réalisé par une réaction de « Click Chemistry », la cycloaddition de Huisguen catalysée par le cuivre I. Les tests antibactériens révèlent l’importance du substituant de l’aryle, l’influence du temps de contact et la pertinence d’utiliser des mélanges de matériaux. L’activité antibactérienne observée sur les fibres de la pâte thermomécanique est meilleure dans les deux parties. Les différents résultats obtenus sont décrits dans ce manuscrit. / The contamination of surfaces by bacteria and the emergence of antimicrobial resistant strains are very worrying problems in different areas such as hospital and food. This contamination begins with the adhesion of pathogenic bacteria on a surface until the formation of biofilms. These biofilms contribute to the emergence of resistances of certain bacterial strains to conventional treatments. To answer these problems of surface contamination, this thesis work focuses on the development of new antibacterial materials based on pulp fibers. In the first part, we focused on the development of an antibacterial paper by grafting, via triazine link, two essential oil compounds, thymol and carvacrol, known for their antibacterial activities. The microbiological evaluation of the developed materials against the two bacterial strains tested, E. coli and S. aureus, showed a bacteriostatic effect. These materials block the bacterial growth thus preventing the biofilms formation. Synergy between thymol and carvacrol grafted onto paper has also been shown. In a second part, our study focused on the development of an antibacterial paper that acquires its activity only after the grafting and formation of "aryl-1,2,3-triazole", the active motif. The grafting is carried out by a reaction of "Click Chemistry", the copper (I)-catalyzed Azide Alkyne Cycloaddition. The antibacterial tests reveal the importance of the aryl substituent, the influence of the contact time and the relevance of using mixtures of materials. The antibacterial activity observed on the thermomechanical pulp fibers is better in both parts. The different results obtained are described in this manuscript.

In terms of morbidity and mortality, dengue is considered the important human viral disease transmitted by mosquitoes, caused by virus of the family Flaviridae, one of the main problems in public health in the world, due to the great potential for the development of several forma and lethal disease. Another problem about dengue control is the population of Aedes aegypti resistant to toxicity of chemical insecticides, such as temephos. This work involved the formulation and characterization of inclusion complex of essential of Lippia gracilis (LGEO), with potent larvicidal effect against Aedes Aegypti Linnaeus larvae in β-cyclodextrin (β-CD), aiming to improve oil solubility in aqueous serving as a strategy for obtaining new products larvicides. Inclusion complexes were obtained by kneading and coevaporation. The complexes obtained by paste and co-evaporation was assessed the influence of the co-solvent used to preparation the complexes by paste with water (PMW), kneading with ethanol / water (PME) co-evaporation with water (COW) and co-evaporation of ethanol / water (COE) methods. The chemical LGEO constituents were identified by gas chromatography coupled to mass spectrometer (GC / MS) where the complex (PMW) was superior in the inclusion of carvacrol with a content of 15.25% compared with the another techniques preparation. The complexes were characterized by thermogravimetry (TG / DTG), the moisture content by Karl-Fischer, differential scanning calorimetry (DSC), X-ray diffraction (X-RD), spectroscopy in the infrared with Fourier transform ( FTIR) and ultra violet (UV-vis), scanning electron microscopy (SEM) and solubility tests. According to the results obtained by GC / MS the major constituent of OELG was carvacrol (46.7%). In TG / DTG and DSC revealed the complex displacement in the range of thermal degradation of the oil to higher temperature ranges indicating a gain of thermal stability. In the analyzes of (X-RD) of the complexes showed the loss of crystalline order compared to pure β-CD. The scanning electron microscopy (SEM) showed changes in particle shapes and morphology of the complex when compared with pure β-CD. The larvicidal assay against Aedes aegypti larvae were carried out with the Rockefeller strain in the third stage (L3), and mortality was observed after 24 hs. The phase solubility diagram at different temperatures, the values of the stability complex of the constant (PMW), was demonstrates be stable and water soluble the interaction complex OELG / β-CD. Aedes aegypti larvae were susceptible to the composition of OELG, carvacrol and complex (PMW) using the method of Probit analysis. The larvicidal activity of the complex (PMW) LC50 was 33 ppm, close to that of pure oil LC50 39 ppm and may be a suitable alternative to biolarvicida dengue control programs. / Em termos de morbidade e mortalidade, a dengue é considerada atualmente a mais importante doença viral humana transmitida por mosquitos, causada por vírus da família Flaviridae, transmitida, no Brasil, através do mosquito Aedes aegypti Linnaeus, sendo um dos principais problemas de saúde pública no mundo, devido ao grande potencial para o desenvolvimento de formas graves e letais da doença. Outro problema enfrentado no controle da dengue é à existência de populações de Aedes aegypti resistentes a toxicidade de inseticidas químicos, como o temefós. Este trabalho envolveu a obtenção e caracterização do complexo de inclusão entre óleo essencial de Lippia gracilis (OELG), com atividade larvicida contra as larvas de Aedes aegypti em β-ciclodextrina (β-CD), objetivando melhorar a solubilidade do óleo em meio aquoso servindo de estratégia para obtenção de novos produtos larvicidas. Para os complexos obtidos por malaxagem e co-evaporação foi avaliada a influência do uso de um co-solvente na preparação dos complexos, pelos métodos malaxagem com água (MAH), malaxagem com etanol/água (MAE), co-evaporação com água (COH) e co-evaporação etanol/água (COE). A determinação do teor do OELG foi obtido por cromatografia gasosa acoplada a espectrômetro de massa (CG/EM), onde o complexo (MAH) mostrou-se superior na inclusão do carvacrol, com teor de 15,25 %, em relação as outras técnicas de preparação. Os complexos foram caracterizados por termogravimetria (TG/DTG), determinação do teor de umidade por Karl-Fischer, calorimetria exploratória diferencial (DSC), difratometria de raios-X (DRX), espectroscopia de absorção na região do infravermelho com transformada de Fourier (FTIR) e na região do ultra violeta (UV-vis), microscopia eletrônica de varredura (MEV) e ensaios de solubilidade. Os ensaios larvicidas contra as larvas de Aedes aegypti foram realizados com as larvas da linhagem Rockefeller no terceiro estádio (L3), sendo verificado a mortalidade após 24 horas. De acordo com os resultados obtido por CG/EM o principal constituinte do OELG, foi o carvacrol (46,7%). Nas curvas TG/DTG e DSC dos complexos revelaram deslocamento no intervalo de degradação térmica do óleo para faixas de temperaturas mais elevadas indicando um ganho de estabilidade térmica. Nas análises de (DR-X) dos complexos demonstraram redução do grau de cristalinidade, quando comparado a β-CD pura. Através da microscopia eletrônica de varredura (MEV) pode-se observar mudanças na morfologia do complexo, quando comparado com a β-CD pura. No estudo do diagrama de fases em diferentes temperaturas, os valores da constante de associação do complexo (MAH), demonstrou serem interações estáveis e que o complexo OELG/β-CD e solúvel em água. As larvas de Aedes Aegypti foram suscetíveis à composição do OELG, carvacrol e do complexo (MAH), utilizando o método de análise Probit. A atividade larvicida do complexo (MAH) foi de CL50 33 ppm, próxima ao do óleo puro de CL50 39 ppm, podendo ser uma alternativa biolarvicida adequada aos programas de controle da dengue.

Es wurden zwei Feldstudien durchgeführt, um zu ermitteln, ob der Verlauf von Eimeria spp.-Infektionen bei Zicklein bei natürlicher Exposition durch die Beimischung von Zusätzen mit ätherischen Ölen (EIMERICOX®= EO1, NEXT Enhance® 200 = EO2) in den Milchaustauscher (MAT) oder das Einbringen eines kommerziellen Pulvers (Stalosan® F = STA) in die Einstreu beeinflusst werden kann. Als Kriterien zur Bewertung der Wirksamkeit dienten die Ausscheidung von Eimeria-Oozysten, die Kotkonsistenz und die klinische Symptomatik sowie die Gewichtsentwicklung. Des Weiteren wurde das Spektrum der caprinen Eimeria-Spezies in der untersuchten Population erfasst. In Studie 1 wurden in einem niederländischen Aufzuchtbetrieb 45 Zicklein verschiedener Herkunft zu je 15 Studientieren auf die Gruppen A1 (Kontrolle), B1 (EO1) und C1 (STA) aufgeteilt. In Studie 2 wurden in einem deutschen Milchziegenbetrieb 45 Zicklein aus betriebseigener Nachzucht in analoger Weise auf die Gruppen A2 (Kontrolle), B2 (EO1) und C2 (EO2) randomisiert. Die Zusätze auf Basis ätherischer Öle wurden während des Studienzeitraums kontinuierlich dem MAT beigemischt (EO1: 4 g/kg MAT; EO2: 125 g/t MAT) und über die MAT-Tränke ad libitum angeboten. Das Einstreupulver wurde zweimal wöchentlich in die Einstreu eingebracht. Insgesamt wurden in Studie 1 (53 Studientage) 484 Kotproben und in Studie 2 (56 Studientage) 847 Kotproben gesammelt, hinsichtlich ihrer Konsistenz bewertet und parasitologisch sowie teilweise differentialdiagnostisch untersucht. Die koproskopischen Untersuchungen umfassten die quantitative Bestimmung der ausgeschiedenen Oozysten pro Gramm Kot (OpG) und die Speziesdifferenzierung. Ferner wurden die Zicklein in Studie 1 am Studientag (ST) 0 und ST 35 und in Studie 2 am ST 0, ST 14, ST 35 und ST 56 gewogen und es erfolgte eine tägliche Kontrolle des Allgemeinbefindens. In beiden Studien wurden natürliche Infektionen der Zicklein mit Eimeria spp. beobachtet. In Studie 1 konnte bei 20 (44,4 %) und in Studie 2 bei 37 (82,2 %) der 45 Studientiere eine Oozystenausscheidung nachgewiesen werden. Die Befallsraten in den Kontrollgruppen (A1 und A2) lagen bei 73,3%in Studie 1 und 93,3%in Studie 2, was einen hohen bis sehr hohen Infektionsdruck anzeigt. Klinisch manifeste Eimeriosen mit Diarrhoe konnten während beider Studien nicht beobachtet werden. In Studie 2 wurde zwar bei 8,1 bis 8,5% der Kotproben in den drei Gruppen Durchfall festgestellt, dieser stand aber in keinem Zusammenhang mit einer Oozystenausscheidung von Eimeria spp. im Allgemeinen oder der pathogenen Spezies E. ninakohlyakimovae im Besonderen. Die Infektionen verliefen also ausnahmslos subklinisch, dennoch ist angesichts der hohen Befallsraten und der Präsenz pathogener Arten das Risiko eines Ausbruchs klinischer Kokzidiosen gegeben. Insgesamt handelte es sich im Falle von Oozystenausscheidung mehrheitlich um Polyinfektionen (Studie 1: 90,0 %; Studie 2: 94,4 %). In beiden Beständen wurde die Präsenz folgender neun Eimeria (E.) spp. nachgewiesen: E. alijevi, E. ninakohlyakimovae, E. hirci, E. arloingi, E. jolchijevi, E. christenseni, E. caprina, E. caprovina. Die höchste Prävalenz in den positiven Proben zeigten (in beiden Studien) E. arlongi (Studie 1: 52,5 %; Studie 2: 78,1 %) und E. ninakohlyakimovae (Studie 1: 55,9 %; Studie 2: 47,8 %). Am seltensten traten E. caprovina (5,1 %) und E. apsheronica (3,4 %) in Studie 1 sowie E. caprina (3,9 %) und E. caprovina (1,7 %) in Studie 2 auf. Die Intensität der Oozystenausscheidung bewegte sich bei 78,0% der positiven Proben in Studie 1 und 88,9% der positiven Proben in Studie 2 im niedrigen bis moderaten Bereich von 50 bis 10 000 OpG. Eine Oozystenausscheidung von mehr als 100 000 OpG (sehr hoher Bereich) trat in Studie 2 bei 1,1% der positiven Proben auf. Die Maxima in Einzelkotproben lagen in Studie 1 bei 67 000 OpG und in Studie 2 bei 157 000 OpG. Anlässlich der Prüfung möglicher Auswirkungen von STA fiel auf, dass gegen Ende des Studienzeitraums in der Gruppe C1 weniger (Re-) Infektionen im Vergleich zur Kontrolle auftraten. Auch die OpG-Werte erwiesen sich bei Auswertung der kumulierten Daten für den gesamten Studienzeitraum und den Zeitraum ab ST 47 sowie am ST 51 in Gruppe C1 als signifikant geringer als in Gruppe A1. Hinsichtlich der Untersuchung von Effekten durch den Einsatz von EO1 und EO2 wurden bei der Analyse von Befallsextensität und Ausscheidungsintensität in Studie 1 in Gruppe B1 niedrigere Befallsraten festgestellt und bei der Auswertung der über den Studienzeitraum kumulierten Daten und gegen Ende der Studie am ST 47, ST 49, ST 51 und ST 52 signifikant weniger positive Kotproben und geringere OpG- Werte als in Gruppe A1 ermittelt. Bei den Befallsraten und der kumulierten Anzahl positiver Proben konnten solche Effekte in Studie 2 nicht reproduziert werden. Gleichwohl zeigte sich am ST 35 und ST 37 in Gruppe B2 eine signifikant niedrigere Eimeria-Prävalenz als in Gruppe A2 und die Intensität der Oozystenausscheidung war in Gruppe B2 am ST 37 und in Gruppe C2 für den Zeitraum ab ST 26 sowie am ST 35, ST 37 und ST 42 signifikant geringer als in Gruppe A2. Bei den mittleren Zunahmen der Körpergewichte fielen stets leichte Gruppenunterschiede zu Gunsten der behandelten Gruppen auf (Gruppe A1: 4,33 kg, Gruppe B1: 5,58 kg, Gruppe C1: 4,55 kg und Gruppe A2: 9,70 kg, Gruppe B2: 10,39 kg, Gruppe C2: 10,74 kg) sie waren jedoch nicht statistisch signifikant. Der subjektive Eindruck einer besseren körperlichen Kondition und Gesamtentwicklung der EO1-behandelten Zicklein wurde dagegen in beiden Studien für einige Parameter und Studientage als signifikant bestätigt. Am Ende der Studien fielen zwölf der 14 Studientiere in Gruppe B1 (Studie 1) und zehn der 14 Studientiere in Gruppe B2 (Studie 2) durch einen exzellenten Entwicklungszustand auf, während dies in den Kontrollgruppen auf drei von zwölf Zicklein in Gruppe A1 (Studie 1) und fünf von 15 Zicklein in Gruppe A2 (Studie 2) zutraf. Der Einsatz von Stalosan® F in Studie 1 schien sich günstig auf den Verlauf der Kokzidiose auszuwirken. Der Eindruck einer kokzidioziden Wirkung sollte jedoch mittels weiterer Studien überprüft werden. Durch die Anwendung der Zusätze auf Basis ätherischer Öle, EIMERICOX® und NEXT Enhance® 200, konnte die Oozystenausscheidung unter subklinischen Bedingungen zwar begrenzt beeinflusst werden, bei einer massiven Exposition ist allerdings kaum davon auszugehen, dass eine Behandlung mit diesen untersuchten ätherischen Ölen eine ausreichend protektive Wirkung hätte, um Eimeriosen effektiv kontrollieren zu können. Bei einer geringen bis moderaten Expositionssituation könnten die geprüften alternativen Maßnahmen ergänzend zu weiteren prophylaktischen Maßnahmen (insbesondere im Haltungs- und Hygienemanagement) dem Aufbau eines kritischen Infektionsdrucks entgegen wirken und somit ein opportunes Anwendungsgebiet finden.

Estudos revelam que o extrato de orégano possui muitas funções na fisiologia animal. Resultados positivos de desempenho e produção foram encontrados em aves e suínos. Em ruminantes, alguns estudos foram conduzidos de forma a avaliar seus efeitos no rúmen, mas são inexistentes os que abordam seu efeito sobre o comportamento animal. No extrato de orégano, há predominância dos óleos essenciais carvacrol e timol. Este estudo objetivou avaliar as modificações no consumo e no comportamento ingestivo e social de novilhas recebendo diferentes doses de extrato de orégano (EO) na dieta. Foram utilizadas 34 novilhas holandesas mantidas em pastejo, com idade de 18,4 ± 4,2 meses e peso 424,2 ± 94,4 Kg, distribuídos em quatro tratamentos: C = controle - sem adição do extrato de orégano; EO 2,5 = 2,5 gramas; EO 5 = 5,0 gramas e EO 7,5 = 7,5 gramas/dia de extrato de orégano. O experimento teve duração de 77 dias, sendo os primeiros 21 de adaptação e constou de avaliações comportamentais focais feitas visualmente em um período de 8 horas, das 10:00 até às 18:00 horas (GMT -3:00). Os valores de consumo e as variáveis do comportamento ingestivo e mudanças posturais foram submetidas à análise de variância e regressão (PROC MIXED e PROC REG), adotou-se o nível 5% de significância. Consideraram-se os efeitos de dia, tratamento, interação tratamento e dia, as medidas realizadas no período pré-experimental foram utilizadas como covariáveis. As variáveis do comportamento social foram transformadas em respostas binárias e submetidas à análise logística (PROC LOG) pelo SAS Enterprise Guide 5.1. Não houve alteração no consumo e no comportamento ingestivo dos animais. Houve interação tratamento x dia para tempo gasto em decúbito, havendo um aumento linear no 46° dia após o início da suplementação. A chance de ocorrência de eventos de dominância aumentou em 1,12 vezes para cada 2,5 gramas de extrato de orégano adicionado à dieta (P=0,0496), enquanto a chance de ocorrência de eventos de submissão não foi alterada. A inclusão de até 7,5 gramas/dia de extrato de orégano não altera o consumo e comportamento ingestivo, porém aumenta os eventos de dominância e pode aumentar o tempo em decúbito de novilhas da raça Holandesa. / Oregano extract has many functions in animal’s physiology. Positive results on performance and production were found in poultry and swine. In ruminants, studies were conducted to evaluate the effects on the rumen metabolism, but few have addressed the effect on animal behavior. In the oregano extract, carvacrol and timol essential oils are prevalent. This study aimed to evaluate changes in ingestive and social behavior in heifers receiving oregano extract (OE) in the diet. Thirty-four Holstein heifers kept in grazing, aged between 18.4 ± 4.2 months and with 424.2 ± 94.4 kg BW were distributed into four treatments: C = control - without addition of oregano extract; OE 2.5 = 2.5 grams; 5 OE = 5.0 grams and OE 7.5 = 7.5 grams/day of oregano extract. The experiment lasted 77 days, with 21 days of adaptation. Behavior evaluation was performed through visual and focal observations of the animals during 8 hours for the day, from 10:00 to 18:00 h (GMT -3:00). Data about intake and ingestive behavior and change in posture were submitted to analysis of variance and regression (PROC MIXED e PROC REG), significance was settled at 0.05. Testing the effects of day, treatment, interaction between treatment and day, and the pre experimental measurements were used for adjustment by covariance. The variables of social behavior were transformed into binary answers and submitted to logistic regression (PROC LOGISTIC) using the SAS Enterprise Guide 5.1. No differences in the feed intake and ingestive behavior of the heifers were detected between treatments. etected. On the 46° day after the beginning of OE supplementation, time spent lying increased with OE addition into the diet. The risk of occurrence of dominance events increased by 1.12 times for each 2.5 grams of the oregano extract added in the diet (P=0.0496), while OE did not change the risk of occurrence of submission events. Inclusion of up to 7.5 grams/day of oregano extract does not change feed intake and ingestive behavior, however increases the chance of occurrence of dominance events and may increase the time spent lying of Holstein heifers.

Estudos revelam que o extrato de orégano possui muitas funções na fisiologia animal. Resultados positivos de desempenho e produção foram encontrados em aves e suínos. Em ruminantes, alguns estudos foram conduzidos de forma a avaliar seus efeitos no rúmen, mas são inexistentes os que abordam seu efeito sobre o comportamento animal. No extrato de orégano, há predominância dos óleos essenciais carvacrol e timol. Este estudo objetivou avaliar as modificações no consumo e no comportamento ingestivo e social de novilhas recebendo diferentes doses de extrato de orégano (EO) na dieta. Foram utilizadas 34 novilhas holandesas mantidas em pastejo, com idade de 18,4 ± 4,2 meses e peso 424,2 ± 94,4 Kg, distribuídos em quatro tratamentos: C = controle - sem adição do extrato de orégano; EO 2,5 = 2,5 gramas; EO 5 = 5,0 gramas e EO 7,5 = 7,5 gramas/dia de extrato de orégano. O experimento teve duração de 77 dias, sendo os primeiros 21 de adaptação e constou de avaliações comportamentais focais feitas visualmente em um período de 8 horas, das 10:00 até às 18:00 horas (GMT -3:00). Os valores de consumo e as variáveis do comportamento ingestivo e mudanças posturais foram submetidas à análise de variância e regressão (PROC MIXED e PROC REG), adotou-se o nível 5% de significância. Consideraram-se os efeitos de dia, tratamento, interação tratamento e dia, as medidas realizadas no período pré-experimental foram utilizadas como covariáveis. As variáveis do comportamento social foram transformadas em respostas binárias e submetidas à análise logística (PROC LOG) pelo SAS Enterprise Guide 5.1. Não houve alteração no consumo e no comportamento ingestivo dos animais. Houve interação tratamento x dia para tempo gasto em decúbito, havendo um aumento linear no 46° dia após o início da suplementação. A chance de ocorrência de eventos de dominância aumentou em 1,12 vezes para cada 2,5 gramas de extrato de orégano adicionado à dieta (P=0,0496), enquanto a chance de ocorrência de eventos de submissão não foi alterada. A inclusão de até 7,5 gramas/dia de extrato de orégano não altera o consumo e comportamento ingestivo, porém aumenta os eventos de dominância e pode aumentar o tempo em decúbito de novilhas da raça Holandesa. / Oregano extract has many functions in animal’s physiology. Positive results on performance and production were found in poultry and swine. In ruminants, studies were conducted to evaluate the effects on the rumen metabolism, but few have addressed the effect on animal behavior. In the oregano extract, carvacrol and timol essential oils are prevalent. This study aimed to evaluate changes in ingestive and social behavior in heifers receiving oregano extract (OE) in the diet. Thirty-four Holstein heifers kept in grazing, aged between 18.4 ± 4.2 months and with 424.2 ± 94.4 kg BW were distributed into four treatments: C = control - without addition of oregano extract; OE 2.5 = 2.5 grams; 5 OE = 5.0 grams and OE 7.5 = 7.5 grams/day of oregano extract. The experiment lasted 77 days, with 21 days of adaptation. Behavior evaluation was performed through visual and focal observations of the animals during 8 hours for the day, from 10:00 to 18:00 h (GMT -3:00). Data about intake and ingestive behavior and change in posture were submitted to analysis of variance and regression (PROC MIXED e PROC REG), significance was settled at 0.05. Testing the effects of day, treatment, interaction between treatment and day, and the pre experimental measurements were used for adjustment by covariance. The variables of social behavior were transformed into binary answers and submitted to logistic regression (PROC LOGISTIC) using the SAS Enterprise Guide 5.1. No differences in the feed intake and ingestive behavior of the heifers were detected between treatments. etected. On the 46° day after the beginning of OE supplementation, time spent lying increased with OE addition into the diet. The risk of occurrence of dominance events increased by 1.12 times for each 2.5 grams of the oregano extract added in the diet (P=0.0496), while OE did not change the risk of occurrence of submission events. Inclusion of up to 7.5 grams/day of oregano extract does not change feed intake and ingestive behavior, however increases the chance of occurrence of dominance events and may increase the time spent lying of Holstein heifers.

Estudos revelam que o extrato de orégano possui muitas funções na fisiologia animal. Resultados positivos de desempenho e produção foram encontrados em aves e suínos. Em ruminantes, alguns estudos foram conduzidos de forma a avaliar seus efeitos no rúmen, mas são inexistentes os que abordam seu efeito sobre o comportamento animal. No extrato de orégano, há predominância dos óleos essenciais carvacrol e timol. Este estudo objetivou avaliar as modificações no consumo e no comportamento ingestivo e social de novilhas recebendo diferentes doses de extrato de orégano (EO) na dieta. Foram utilizadas 34 novilhas holandesas mantidas em pastejo, com idade de 18,4 ± 4,2 meses e peso 424,2 ± 94,4 Kg, distribuídos em quatro tratamentos: C = controle - sem adição do extrato de orégano; EO 2,5 = 2,5 gramas; EO 5 = 5,0 gramas e EO 7,5 = 7,5 gramas/dia de extrato de orégano. O experimento teve duração de 77 dias, sendo os primeiros 21 de adaptação e constou de avaliações comportamentais focais feitas visualmente em um período de 8 horas, das 10:00 até às 18:00 horas (GMT -3:00). Os valores de consumo e as variáveis do comportamento ingestivo e mudanças posturais foram submetidas à análise de variância e regressão (PROC MIXED e PROC REG), adotou-se o nível 5% de significância. Consideraram-se os efeitos de dia, tratamento, interação tratamento e dia, as medidas realizadas no período pré-experimental foram utilizadas como covariáveis. As variáveis do comportamento social foram transformadas em respostas binárias e submetidas à análise logística (PROC LOG) pelo SAS Enterprise Guide 5.1. Não houve alteração no consumo e no comportamento ingestivo dos animais. Houve interação tratamento x dia para tempo gasto em decúbito, havendo um aumento linear no 46° dia após o início da suplementação. A chance de ocorrência de eventos de dominância aumentou em 1,12 vezes para cada 2,5 gramas de extrato de orégano adicionado à dieta (P=0,0496), enquanto a chance de ocorrência de eventos de submissão não foi alterada. A inclusão de até 7,5 gramas/dia de extrato de orégano não altera o consumo e comportamento ingestivo, porém aumenta os eventos de dominância e pode aumentar o tempo em decúbito de novilhas da raça Holandesa. / Oregano extract has many functions in animal’s physiology. Positive results on performance and production were found in poultry and swine. In ruminants, studies were conducted to evaluate the effects on the rumen metabolism, but few have addressed the effect on animal behavior. In the oregano extract, carvacrol and timol essential oils are prevalent. This study aimed to evaluate changes in ingestive and social behavior in heifers receiving oregano extract (OE) in the diet. Thirty-four Holstein heifers kept in grazing, aged between 18.4 ± 4.2 months and with 424.2 ± 94.4 kg BW were distributed into four treatments: C = control - without addition of oregano extract; OE 2.5 = 2.5 grams; 5 OE = 5.0 grams and OE 7.5 = 7.5 grams/day of oregano extract. The experiment lasted 77 days, with 21 days of adaptation. Behavior evaluation was performed through visual and focal observations of the animals during 8 hours for the day, from 10:00 to 18:00 h (GMT -3:00). Data about intake and ingestive behavior and change in posture were submitted to analysis of variance and regression (PROC MIXED e PROC REG), significance was settled at 0.05. Testing the effects of day, treatment, interaction between treatment and day, and the pre experimental measurements were used for adjustment by covariance. The variables of social behavior were transformed into binary answers and submitted to logistic regression (PROC LOGISTIC) using the SAS Enterprise Guide 5.1. No differences in the feed intake and ingestive behavior of the heifers were detected between treatments. etected. On the 46° day after the beginning of OE supplementation, time spent lying increased with OE addition into the diet. The risk of occurrence of dominance events increased by 1.12 times for each 2.5 grams of the oregano extract added in the diet (P=0.0496), while OE did not change the risk of occurrence of submission events. Inclusion of up to 7.5 grams/day of oregano extract does not change feed intake and ingestive behavior, however increases the chance of occurrence of dominance events and may increase the time spent lying of Holstein heifers.

Abiotic factors may interfere in production of plant secondary metabolites and consequently influence the bioactivity of these compounds on insect pests. Thus, this study aimed to evaluate the influence of different agricultural practices (irrigation, mineral fertilizer and organic manure) in chemical composition of the essential oil (EO) of Lippia gracilis and its effects on the maize weevil Sitophilus zeamais.The chemical compositions of EO from plants of L. gracilis cultured on different treatments were analyzed using GC-MS and GC-FID. The carvacrol was the major component of EO in different agricultural practices followed by thymol and ñ-cymene.These three monoterpenes were used in subsequent bioassays. Cultivation practices did not significantly affect the composition of the OEs of L. gracilis. However, small variations in the composition of EOs resulted in different biological responses to S. zeamais. All EOs of L. gracilis and its main monoterpenes applied topically showed low toxicity on S. zeamais after 84 hours exposure at a dose 10 mg g-1. However, these same compounds were highly toxic to S. zeamais by fumigation. The time required to cause 50% mortality of the pest was 28.4 hours on average, ranging from 17.3 to 40.8 hours between treatments. The toxic action of carvacrol, thymol and EOs of L. gracilis (no irrigation/fertilization mineral and 40 t ha-1 organic fertilizer) were the fastest, LT50=18,3. The magnitude of the variation of CL50 and CL99 were 1.9 and 2.1 times. The concentration needed to cause 50 to 99% mortality of the pest was on average 69 and 260 ìl l-1, respectively.Considering the LC50 and LC99, the more toxic OEs of L. gracilis were obtained from plants grown in the absence of mineral fertilizer and in the presence of organic manure with of 20, 40 and 60 t ha-1 (no irrigation) and 40 and 60 t ha-1 (with irrigation). Much of the activity of the EOs of L. gracilis is due to the major compound carvacrol. It took only 27.2 and 92.2 l ìl l-1 to cause 50% and 99 % mortality of adult S. zeamais. The toxic activity of this monoterpene was still one of the fastest (LT50 = 17.3 and LT99 = 70.8 hours). Already the thymol and ñ-cymene awere compounds of low toxicity to adult S. zeamais (LC50= 4,4 e 26,1 respectively). Generally, the EOs of L. gracilis and the monoterpenes applied at lower concentrations (0.01 and 0.1% v/v) showed a neutral effect (not significant) or attractive (negative and significant) in all periods analyzed. At concentration 10% (v/v) almost all treatments were repellents to S. zeamais at any time analyzed (repellence > 50%). Major indexes repellency (54-86%) were observed in treatments irrigated and with maximum organic manure, at concentrations of 1 and 10% (v/v). Thus, our results show the great potential of EO of L. gracilis and its major compound for the development of new products and programs for use in integrated pest management of stored grain. / Fatores abioticos podem interferir na producao de metabolitos secundarios de plantas e consequentemente influenciar na bioatividade destes compostos sobre insetos-praga. Assim, neste trabalho objetivou-se avaliar a influencia de diferentes praticas agricolas (irrigacao, adubacao quimica e organica) na composicao quimica do oleo essencial (OE) de Lippia gracilis e seus efeitos na mortalidade e repelencia do gorgulho do milho Sitophilus zeamais. As composicoes quimicas dos OEs das plantas de L. gracilis cultivadas sobre diferentes tratamentos foram analisadas usando CG-MS e CG-FID. As praticas de cultivo nao afetaram significativamente a composicao dos OEs de L. gracilis, o carvacrol foi o componente majoritario dos OEs nas diferentes praticas agricolas, seguido pelo timol e Ï-cimeno. Estes tres monoterpenos foram utilizados nos bioensaios posteriores. Pequenas variacoes na composicao dos OEs resultaram em diferentes respostas biologicas de S. zeamais. Todos os OEs de L. gracilis aplicados topicamente mostraram baixa toxicidade sobre S. zeamais apos 84 horas de exposicao na dose 10 mg g-1. Contudo, estes mesmos compostos foram altamente toxicos a S. zeamais por fumigacao. O tempo necessario para causar 50% de mortalidade desta praga foi em media 28,4 horas; variando de 17,3 a 40,8 horas entre os tratamentos. A acao toxica do carvacrol, timol e do OEs de L. gracilis (ausencia de irrigacao/adubacao mineral e 40 t ha-1 de adubacao organica) foram as mais rapidas, TL50=18,3. A magnitude da variacao das CL50 e CL99 foram de 1,9 e 2,1 vezes. A concentracao necessaria para causar 50 e 99% de mortalidade desta praga foi em media 69 e 260 Êl l-1, respectivamente. Considerando as CL50 e CL99, os OEs de L. gracilis mais toxicos foram obtidos de plantas cultivadas na ausencia de adubacao mineral e presenca de adubacao organica com 20, 40 e 60 t ha-1 (sem irrigacao) e 40 e 60 t ha-1 (com irrigacao). Grande parte da atividade dos OEs de L. gracilis deve-se ao composto majoritario carvacrol. Foram necessarios apenas 27,2 e 92,2 Êl l-1 para causar 50 e 99% de mortalidade a adultos de S. zeamais. A atividade toxica deste monoterpeno foi de TL50 = 17,3 e TL99 = 70,8 horas. Ja o timol e Ï-cimeno foram os compostos de menor toxicidade a adultos de S. zeamais (CL50= 4,4 e 26,1 respectivamente). De forma geral, os OEs de L. gracilis e os monoterpenos utilizados nas menores concentracoes (0,01 e 0,1% v/v) apresentaram efeito neutro (nao significativo) ou atrativo (negativo e significativo) em todos os tempos analisados. Ja na concentracao 10% (v/v) quase todos os tratamentos foram repelentes a S. zeamais em qualquer tempo analisado (repelencia > 50%). Maiores indices de repelencia (54-86%) foram observadas nos tratamentos irrigados com maxima adubacao organica e concentracao 1 e 10% (v/v). Assim, nossos resultados mostram o grande potencial do OE de L. gracilis e seu composto majoritario para o desenvolvimento de novos produtos e utilizacao em programas de manejo integrado de pragas de graos armazenados.