Despite significant vector control advancements in the past years, the current malaria trends suggest that new control strategies are urgently required. These new approaches should address the current frontline intervention challenges like increasing insecticide resistance in mosquitoes and residual transmission issues. Insecticide-treated livestock (ITL) is one of the novel potential strategies to overcome the above challenges. ITL involves treating livestock near humans with an insecticide like ivermectin (IVM) to kill zoophagic malaria vectors. However, ivermectin pharmacokinetics data suggests that most IVM-administered drugs remain intact, and more than 90 % of this drug is eliminated in feces. Biological metabolites: 3′′-O-demethylivermectin (3DI) and 24-hydroxymethyl ivermectin (24OHI) are also excreted in feces. Therefore, using manure from treated cattle as fertilizers contaminates the soil, ground, and surface water with IVM or its metabolites through leaching and hydraulic water flow affecting the soil and aquatic ecosystems. Contemplating the contamination impacts, these drugs' environmental fate and effects could be regarded before massive IVM applications. Many researchers have tried to address this subject in temperate regions compared to the tropics, where IVM is urgently needed. Regional discrepancies such as soil types and climate can independently and dependently determine the fate and impact of ivermectin. Our research investigates the environmental fate of IVM and its primary biological metabolites. Laboratory and field studies in Tanzania and Virginia were conducted to simulate the difference between tropical and temperate climates. Soil and soil-manure mixture spiked with IVM were layered into two 5 mm layers in columns exposed to natural sunlight. The remaining IVM and its primary metabolite were quantified using Liquid Chromatography with a tandem mass spectrometry detector (LC-MS/MS. These compounds degraded up to 1.5 times faster in Tanzania than in Virginia, depending on temperature, soil depths and type, organic matter, and soil moisture. When IVM is subcutaneously injected into cattle, drug residues and metabolites: 3DI and 24OHI are excreted in feces following a positive skewed Poisson distribution profile. IVM, 3DI, and 24OH were found to degrade rapidly when cattle pats when exposed to the field. Since we conducted our study in the Summer, no IVM or its metabolites leached into the soil beneath. The obtained half-lives suggest that ivermectin's massive drug administration has little to worry about, primarily when the dung from treated cattle is spread into the field in thin layers in the Summer before farm application. / Doctor of Philosophy / Despite significant vector control advancements in the past years, the current malaria trends suggest that new control strategies are urgently required. These new approaches should address the current frontline intervention challenges like increasing drug resistance in mosquitoes and residual transmission issues. Treating cattle with ivermectin is one of the novel potential strategies to overcome the above challenges. This strategy is effective because the amount of ivermectin (IVM) found in the blood of treated cattle is enough to kill malaria vectors feeding on them. However, the literature suggests that most IVM-administered drugs remain intact, and more than 90 % of this drug is eliminated in feces. Metabolite bioproducts: 3″-O-demethylivermectin (3DI) and 24-hydroxymethyl ivermectin (24OHI) are also excreted in feces. Therefore, using manure from treated cattle as fertilizers contaminates the soil, ground, and surface water with IVM or its metabolites through leaching and hydraulic water flow affecting the soil and aqua ecosystems. Contemplating the contamination impacts, these drugs' environmental fate and effects could be regarded before massive IVM applications. Many researchers have tried to address this subject in temperate regions compared to the tropics, where IVM is more needed. Regional discrepancies such as soil types and climate can independently and dependently determine the fate and impact of ivermectin. Our research investigates the environmental fate of IVM and its primary bioproducts. Laboratory and field studies in Tanzania and Virginia were conducted to simulate the difference between tropical and temperate climates. Soil and soil-manure mixture spiked with IVM were layered into two 5 mm layers in columns exposed to natural sunlight. The remaining IVM and its primary metabolite were quantified on the appropriate instrument. These compounds degraded up to 1.5 times faster in Tanzania than in Virginia, depending on temperature, soil depths and type, organic matter, and soil moisture. When IVM drug is injected into cattle, the intact drug and its bioproducts: 3DI and 24OHI, are eliminated in feces following a favorable skewed normal distribution profile. IVM, 3DI, and 24OH were found to degrade rapidly when cattle pats when exposed to the field. Since we conducted our study in the Summer, no IVM or its bioproducts leached into the soil beneath. The obtained data suggest that ivermectin's massive drug administration has little to worry about, primarily when the dung from treated cattle is spread into the field in thin layers in the Summer before farm application.

Les infections par les nématodes gastro-intestinaux entrainent des baisses en productions animales et des pertes économiques majeures pour les éleveurs. Les lactones macrocycliques (LMs) sont parmi les antiparasitaires les plus utilisés dans la lutte contre les nématodes gastro-intestinaux en médecine vétérinaire. De part une utilisation intensive, des résistances aux LMs chez les parasites gastro-intestinaux se sont développées au sein des élevages du monde entier mettant en péril l'efficacité thérapeutique de ces molécules. Par ailleurs, le développement de nouveaux antiparasitaires est limité. Ainsi, un des enjeux pour assurer le contrôle de ces parasites est de ralentir les phénomènes de résistance aux LMs afin de prolonger leur efficacité. Le succès d'une telle stratégie repose sur les connaissances précises des mécanismes impliqués dans la résistance. Parmi eux, la modulation des systèmes de détoxification est décrite lors de phénomènes de résistance aux LMs. Au cours de ces travaux, nous avons étudiés la régulation des systèmes de détoxification, en réponse à l'ivermectine chez l'hôte. Nous avons montré, en comparaison à une administration unique, qu'une administration répétée d'ivermectine par voie orale chez la souris est responsable de l'induction de l'expression de certains gènes transporteurs ABC et cytochromes impliqués dans son métabolisme. Ceci entraîne, à la fois, une diminution de la concentration de la molécule parentale et une augmentation de la teneur de son métabolite principal dans le plasma et l'intestin. Ensuite, nous avons étudié l'implication des mécanismes de régulation des systèmes de détoxification, et notamment les récepteurs nucléaires, dans la tolérance à l'ivermectine chez le nématode C.elegans. Nous avons montré que le récepteur nucléaire nhr-a est important pour la tolérance et le développement de la résistance à l'ivermectine. Enfin, nous avons étudié l'impact d'inhibiteurs des transporteurs ABC sur l'efficacité de l'ivermectine. Nous avons mis en évidence la capacité de certains flavonoïdes et de l'ivermectine aglycone à potentialiser l'efficacité de l'ivermectine chez le nématode. Une exposition d'ivermectine induit la surexpression des systèmes de détoxification chez l'hôte. Ceci pourrait être la base des mécanismes moléculaires de la résistance chez le nématode. Cibler les systèmes de détoxification ou les mécanismes de résistance, par des inhibiteurs adaptés, représente une stratégie pertinente pour potentialiser l'efficacité de l'ivermectine. / Infections with gastrointestinal nematodes (GINs) in livestock leads to major losses in production and consequently impact economically farmers. Their intensive use has led to widespread anthelmintic resistance which is nowadays the main threat on the sustainable control of GINs in livestock. The development of new anthelmintic is limited due to the cost of such process. Then, the challenge remains in optimizing the use of existing molecules. Therefore, it is urgent to limit and control MLs resistance in order to extend their efficacy and to avoid therapeutic failure. Resistance mechanisms remain to be elucidated. In that context, we investigated regulatory mechanism of detoxification systems of ivermectin implicated in therapeutic efficacy in host and resistance development in nematode. Therapeutic combinations of ivermectin with flavonoïds have been evaluated to potentiate its efficacy in nematode. We showed that repeated oral administration of ivermectin induced gene expression encoding some ABC efflux transporters and cytochromes involved in its metabolism. Compared with single administration, repeated ivermectin administration lowered plasma, liver and intestine drug concentration, while increasing main metabolite content in plasma and intestine. We have also shown that nuclear receptor nhr-a was important for ivermectin tolerance and ivermectin development of resistance in C. elegans. Finally, we demonstrated the ability of the flavonoïd phloretin to potentiate ivermectin efficacy in the nematode C. elegans. Taken together, these data suggest that induction of detoxification systems impact on ivermectin distribution and targeting their regulation could be an appropriate strategy to potentiate ivermectin efficacy in host and to reverse resistance in nematode.

Die zunehmende Ausbreitung des Usutu-Virus in Europa als Ursache für fatale Ausbruchsgeschehen innerhalb der Avifauna, insbesondere unter Sperlingsvögeln (Passeriformes) und Eulenartigen (Strigiformes), stellt in Zusammenhang mit einem neuroinvasiven sowie zoonotischen Potential ein Risiko für die Veterinär- sowie Humanmedizin dar. Trotz dieser Relevanz stehen derzeit keine zugelassenen Therapeutika gegen eine Usutu-Virus-Infektion zur Verfügung. Auf Basis indikationsfremder Substanzen mit pharmakologischer Zulassung im Sinne des drug repositioning sowie auf Grundlage der Gegenregulation viral-induzierter Modulationen des Zellmetabolismus wurde die Identifikation antiviraler Therapieoptionen gegen das Usutu-Virus in vitro angestrebt.:Abkürzungsverzeichnis Abbildungsverzeichnis Tabellenverzeichnis 1 Einleitung 2 Literaturübersicht 2.1 Das Usutu-Virus 2.1.1 Ursprung, Klassifikation und Epidemiologie 2.1.1 Transmissionszyklus und Wirtstropismus 2.1.2 Aufbau des Virions und des Genoms 2.1.3 Veterinärmedizinische Relevanz und pathologische Ausprägung 2.1.4 Humanmedizinische Bedeutung als Zoonose-Erreger 2.1.5 Vergleichende Aspekte zum West-Nil-Virus 2.2 Antivirale Präventions- und Therapieoptionen 2.2.1 Möglichkeiten und Grenzen der Immunprophylaxe 2.2.2 Pharmaka mit potentiell antiviraler Wirkung gegen Flaviviren 2.2.3 Identifizierte Substanzen gegen das Usutu-Virus 2.3 Zelluläre Systeme als antivirale Zielobjekte 2.3.1 Das Interferon-System und seine antivirale Schutzfunktion 2.3.2 Viral-induzierte Modulation des Wirtszellmetabolismus 3 Zielstellungen der Dissertation 4 Material 4.1 Zelllinien 4.2 Viruslinien 4.3 Zellkulturmedien 4.4 Pharmakologische und andere Substanzen 4.5 Chemikalien 4.6 Lösungen und Puffer 4.7 Antikörper 4.8 Reagenzien und Kit-Systeme 4.9 Enzyme und Nukleotide 4.10 Primer 4.11 Verbrauchsmaterialen 4.12 Geräte 4.13 Datenbanken und Software 5 Methodik 5.1 Zellkultivierungstechnik und PBMC-Isolation 5.2 Isolation von Usutu-Virus-Linien aus Gewebeproben in Zellkultur 5.2.1 Aufbereitung aviären Organmaterials 5.2.2 Typisierung von in Deutschland zirkulierenden Usutu-Virus-Linien (2019, 2020) 5.2.3 Virusanzucht in verschiedenen Zellkulturen zur Virusstock-Generierung 5.3 Quantifizierung des extrazellulären Virustiters 5.4 Immunfluoreszenzanalyse 5.5 Präparation pharmakologischer und anderer Substanzen 5.6 Infektionsansätze mit dem Usutu-Virus und WNV 5.7 Durchflusszytometrische Analyse 5.8 Zytotoxizitätsstudien 5.9 Extrazelluläre Fluxanalyse mittels Agilent Seahorse XF-Technologie 5.10 Statistische Analyse 6 Ergebnisse 6.1 Isolation des Usutu-Virus in Zellkultur 6.2 Replikationsdynamik des Usutu-Virus in verschiedenen Zelllinien 6.3 Pharmakologisches Screening zur antiviralen Wirksamkeit gegen das Usutu-Virus 6.4 Identifikation und Charakterisierung der antiviralen Eigenschaften von Ivermectin 6.5 Wirkung von Ivermectin gegen Linie 2 des West-Nil-Virus in einer aviären Zelllinie 6.6 Metabolischer Phänotyp Usutu-Virus-infizierter Zelllinien 6.7 Antivirale Inhibition der Glykolyse durch 2-Deoxy-D-Glukose 6.8 Einfluss von exogenem Interferon auf den Wirtszellmetabolismus unter Infektion 6.9 Auswirkung der Interferon-Rezeptor-Defizienz auf den Metabolismus unter Infektion 7 Diskussion 7.1 Typisierung und Isolation des Usutu-Virus in Zellkultur 7.2 Charakterisierung der zelltypspezifischen Permissivität und Replikationskinetik 7.3 Identifikation antiviral wirksamer Substanzen gegen das Usutu-Virus 7.4 Usutu-Virus-induzierte Modulation des Metabolismus als antiviraler Ansatz 8 Ausblick 9 Zusammenfassung 10 Summary 11 Literaturverzeichnis 12 Anhang 12.1 Zusatzmaterial 12.2 Publikation 1 12.3 Publikation 2 12.4 Publikation 3 12.5 Weitere Veröffentlichungen 13 Danksagung / The emerge of Usutu virus (USUV) in Europe as a causative agent of fatal outbreaks in avifauna, notably in Passeriformes and Strigiformes, as well as its neuroinvasive and zoonotic potential emphasize a considerable risk in veterinary and human medicine. Despite its relevance, recently, no approved drugs against USUV infections are available. The identification of antiviral therapeutic options against USUV in vitro was addressed based on the analysis of approved drugs of other medical indications in terms of drug repositioning and the counteraction of viral-induced alterations of the cellular metabolism.:Abkürzungsverzeichnis Abbildungsverzeichnis Tabellenverzeichnis 1 Einleitung 2 Literaturübersicht 2.1 Das Usutu-Virus 2.1.1 Ursprung, Klassifikation und Epidemiologie 2.1.1 Transmissionszyklus und Wirtstropismus 2.1.2 Aufbau des Virions und des Genoms 2.1.3 Veterinärmedizinische Relevanz und pathologische Ausprägung 2.1.4 Humanmedizinische Bedeutung als Zoonose-Erreger 2.1.5 Vergleichende Aspekte zum West-Nil-Virus 2.2 Antivirale Präventions- und Therapieoptionen 2.2.1 Möglichkeiten und Grenzen der Immunprophylaxe 2.2.2 Pharmaka mit potentiell antiviraler Wirkung gegen Flaviviren 2.2.3 Identifizierte Substanzen gegen das Usutu-Virus 2.3 Zelluläre Systeme als antivirale Zielobjekte 2.3.1 Das Interferon-System und seine antivirale Schutzfunktion 2.3.2 Viral-induzierte Modulation des Wirtszellmetabolismus 3 Zielstellungen der Dissertation 4 Material 4.1 Zelllinien 4.2 Viruslinien 4.3 Zellkulturmedien 4.4 Pharmakologische und andere Substanzen 4.5 Chemikalien 4.6 Lösungen und Puffer 4.7 Antikörper 4.8 Reagenzien und Kit-Systeme 4.9 Enzyme und Nukleotide 4.10 Primer 4.11 Verbrauchsmaterialen 4.12 Geräte 4.13 Datenbanken und Software 5 Methodik 5.1 Zellkultivierungstechnik und PBMC-Isolation 5.2 Isolation von Usutu-Virus-Linien aus Gewebeproben in Zellkultur 5.2.1 Aufbereitung aviären Organmaterials 5.2.2 Typisierung von in Deutschland zirkulierenden Usutu-Virus-Linien (2019, 2020) 5.2.3 Virusanzucht in verschiedenen Zellkulturen zur Virusstock-Generierung 5.3 Quantifizierung des extrazellulären Virustiters 5.4 Immunfluoreszenzanalyse 5.5 Präparation pharmakologischer und anderer Substanzen 5.6 Infektionsansätze mit dem Usutu-Virus und WNV 5.7 Durchflusszytometrische Analyse 5.8 Zytotoxizitätsstudien 5.9 Extrazelluläre Fluxanalyse mittels Agilent Seahorse XF-Technologie 5.10 Statistische Analyse 6 Ergebnisse 6.1 Isolation des Usutu-Virus in Zellkultur 6.2 Replikationsdynamik des Usutu-Virus in verschiedenen Zelllinien 6.3 Pharmakologisches Screening zur antiviralen Wirksamkeit gegen das Usutu-Virus 6.4 Identifikation und Charakterisierung der antiviralen Eigenschaften von Ivermectin 6.5 Wirkung von Ivermectin gegen Linie 2 des West-Nil-Virus in einer aviären Zelllinie 6.6 Metabolischer Phänotyp Usutu-Virus-infizierter Zelllinien 6.7 Antivirale Inhibition der Glykolyse durch 2-Deoxy-D-Glukose 6.8 Einfluss von exogenem Interferon auf den Wirtszellmetabolismus unter Infektion 6.9 Auswirkung der Interferon-Rezeptor-Defizienz auf den Metabolismus unter Infektion 7 Diskussion 7.1 Typisierung und Isolation des Usutu-Virus in Zellkultur 7.2 Charakterisierung der zelltypspezifischen Permissivität und Replikationskinetik 7.3 Identifikation antiviral wirksamer Substanzen gegen das Usutu-Virus 7.4 Usutu-Virus-induzierte Modulation des Metabolismus als antiviraler Ansatz 8 Ausblick 9 Zusammenfassung 10 Summary 11 Literaturverzeichnis 12 Anhang 12.1 Zusatzmaterial 12.2 Publikation 1 12.3 Publikation 2 12.4 Publikation 3 12.5 Weitere Veröffentlichungen 13 Danksagung

Des systèmes injectables de formation in situ ont été utilisés dans les dernières années pour l'obtention de formulations de préparation facile et permettant la libération prolongée de principes actifs. Ces systèmes utilisant des solvants biocompatibles et des polymères biodégradables sont des liquides (solutions ou émulsions) qui une fois injectés dans l'organisme donnent lieu à des implants (ISI) ou à des microparticules (ISM) solides. La formation de ces systèmes est induite par la précipitation du polymère à partir des solutions polymériques qu'ils contiennent lors du contact avec les fluides corporaux aqueux. Dans ce travail, des ISI et des ISM, réalisés à partir des polymères de l'acide lactique et/ou glycolique (PLA et PLGA) et des différents solvants biocompatibles, pour la libération prolongée d?ivermectine (IVM), un principe actif antiparasitaire faiblement biodisponible par la voie orale, ont été développés. Les profils de libération du principe actif in vitro et in vivo à partir de ces systèmes, ont été comparés avec ceux obtenus à partir de microparticules réalisées par la méthode classique dite d'émulsion simple - évaporation de solvant ; il s'agit d'une technique aux multiples étapes, à coût élevé et dont l'utilisation de solvants toxiques la font difficilement industrialisable. La libération du principe actif à partir des microparticules obtenues par émulsion simple/évaporation du solvant a été influencée par la forte interaction du principe actif avec les polymères mais aussi par la porosité. Dans le cas des systèmes in situ, la vitesse de libération d'IVM a été conditionnée par la solubilité dans l'eau du solvant biocompatible sélectionné et par les interactions solvant/polymère. Pour les ISM, des paramètres tels que la nature de la phase externe, aqueuse (ISM-O/W) ou huileuse (ISM-O/O), la solubilité dans l'eau du solvant de la phase interne, l'affinité entre les phases et l'affinité de l'IVM pour chacune des phases, ont déterminé la vitesse de libération du principe actif. La bonne stabilité ainsi que les profils de libération plus prolongés et présentant une faible libération initiale du principe actif in vivo et in vitro, ont montré que les ISI et les ISM réalisés à partir de solvants biocompatibles de faible solubilité dans l'eau tels que la triacetine sont les plus indiqués pour l'encapsulation d'IVM par rapport à ceux plus solubles dans l'eau comme la N-methyl-2-pyrrolidone et la 2-pyrrolidone. Ces systèmes représentent donc une alternative intéressante par rapport aux formulations conventionnelles d'IVM / In situ forming injectable systems have been used in the past years to obtain sustained drug release formulations which are easy to prepare. These systems using biocompatible solvents and biodegradable polymers are liquids (solutions or emulsions) that upon injection on the body lead to solid implants (ISI) or microparticles (ISM). These systems are formed in contact with water body fluids by polymer precipitation from the polymeric solution. In this work, ISI and ISM made from lactide and/or glycolide polymers (PLA and PLGA) and different biocompatible solvents were performed to obtain sustained release of ivermectin (IVM), an antiparasitic drug with a low oral bioavailability. In vitro and in vivo drug release profiles from these systems were compared with those from microparticles obtained by the classical simple emulsion/solvent evaporation method, which is difficult to propose in industry because of its multiple steps, high cost and the solvent toxicity. Drug release from simple emulsion/solvent evaporation microparticles was affected by the strong polymer/drug interactions and porosity. Concerning to in situ forming systems, the rate of IVM release was dependent on solvent water solubility and solvent/polymer interactions. The nature of the external phase, water (ISM-O/W) or oil (ISM-O/O), the water solubility of the solvent in the internal phase, phase affinity and IVM/phase affinity determined drug release from ISM. The good stability, the in vitro and in vivo sustained release and the low burst effect of IVM, indicated that ISI and ISM formulated from low hydrosoluble biocompatible solvents such as triacetin are more appropriated to IVM formulation instead of those based on more hydrophilic solvent (N-methyl-2-pyrrolidone and 2-pyrrolidone). These systems are an interesting alternative to conventional IVM formulations

Made available in DSpace on 2016-04-28T20:16:30Z (GMT). No. of bitstreams: 1 2009 - Clarissa Pimentel de Souza.pdf: 1413726 bytes, checksum: ad7e79be3b961bab7286264dea8d79c1 (MD5) Previous issue date: 2009-04-29 / The mite Demodex canis is a resident of the hair follicles and when overgrowth cause an inflammatory parasitic disease called demodectic mange. The diagnosis is based on the visualization of the mite under microscope, especially by skin scrapings, hair plucks or histopathologic exam. A large number of drugs has been used on the treatment of demodectic mange, demonstrating varied efficacy numbers, side effects that can prohibit the conclusion of treatment and difficulties to owners. The objective of the present study was to evaluate the efficacy of the fluazuron 2.5% pour on and ivermectin on different therapeutic protocols on the treatment of canine demodectic mange. For this, 31 dogs were divided into five groups, the first four with six animals and the last one with seven. All with positive skin scrapings. The dogs were treated on each 14 days, during 84 days. The first group used fluazuron 2.5% pour on at 20mg/kg, the second used this same drug associated to ivermectin 0.5% pour on at 0.6mg/kg and, the third one, only the ivermectin 0,5% pour on. The fourth and fifth groups were treated with long-acting ivermectin 3.15% on subcutaneous administration at 0.6 and 1.5mg/kg, respectively. The evaluation and follow up of treatments were realized through skin scrapings on each 14 days, clinical evaluation with photos of dogs on every visit for a compare of the lesions. And the histopathologic exam at the end of the therapeutic protocol. The success rate was defined as the percentage of dogs on each group with negative skin scrapings. The reduction in mite numbers reached the efficacy levels of 67.66; 88.99; 84.29; 84.90 and 87.86%, for groups 1, 2, 3, 4 e 5, respectively. And the success rates at the end of treatment were 16.67% for the first group and 50% for the other four. By Wilcoxon test the reduction of the infestation on the histopathologic exam, before and after treatment, had no significance for all groups. Remission of lesions did not occur with the dogs of the first group. On groups 2 and 3, on each one was observed two dogs clinically cured and one with an improvement of the lesions. And on groups 4 and 5, 4 and 5 dogs were cured, respectively and also, there was one with an improvement on the lesions. The fluazuron 2.5% pour on did not show efficacy on the treatment of canine demodectic mange. But the ivermectin 0.5% pour on associated or not to fluazuron 2.5% pour on and the ivermectin 3.15% on both dosages, showed good efficacy on the reduction in mite numbers at the end of the protocols. / O ?caro Demodex canis ? um habitante dos fol?culos pilosos, que quando se prolifera causa uma dermatopatia de cunho inflamat?rio denominada sarna demod?cica. O diagn?stico ? feito atrav?s da visualiza??o do parasito sob microscopia ?ptica, especialmente atrav?s do exame parasitol?gico de raspado cut?neo, dos p?los, ou exame histopatol?gico. Muitas drogas t?m sido utilizadas no tratamento da sarna demod?cica, mas demonstrando n?veis de efic?cia variados, ocorr?ncia de efeitos colaterais que impossibilitam a conclus?o da terapia ou empecilhos ? ades?o dos propriet?rios. O objetivo do presente estudo foi avaliar a efic?cia do fluazuron e da ivermectina em diferentes protocolos terap?uticos no controle da sarna demod?cica canina. Foram avaliados 31 c?es divididos em cinco grupos, os quatro primeiros com seis animais e o ?ltimo com sete, positivos para o ?caro D. canis atrav?s do raspado cut?neo. Todos os c?es foram tratados a cada 14 dias, durante 84 dias. No primeiro grupo foi utilizado fluazuron 2,5% pour on na dosagem de 20mg/kg, no segundo este f?rmaco tamb?m foi empregado, mas associado a ivermectina 0,5% pour on na dosagem de 0,6mg/kg e no terceiro, somente a ivermectina 0,5% pour on . Os c?es do quarto e quinto grupos foram tratados com ivermectina 3,15% longa a??o por via subcut?nea nas dosagens de 0,6 e 1,5mg/kg, respectivamente. A avalia??o e acompanhamento do tratamento foram feitos atrav?s dos exames parasitol?gicos de raspado cut?neo a cada 14 dias, da avalia??o cl?nica, inclusive fotografando os c?es para uma melhor compara??o dos quadros lesionais e, do exame histopatol?gico ao final de cada protocolo terap?utico. A taxa de sucesso foi definida pela porcentagem de c?es em cada grupo que apresentaram raspados negativos. A redu??o na contagem no n?mero de ?caros alcan?ou n?veis de efic?cia de at? 67,66; 88,99; 84, 29; 84,90 e 87,86%, nos grupos 1, 2, 3, 4 e 5, respectivamente. E as taxas de sucesso ao final do tratamento foram de 16,67% para o grupo 1 e 50% para os outros quatro. Pelo do teste de Wilcoxon a redu??o da infesta??o atrav?s do exame histopatol?gico antes e depois do tratamento n?o foi significativa para nenhum grupo. Clinicamente, n?o ocorreu remiss?o das les?es nos c?es do primeiro grupo. Nos grupos 2 e 3, em cada um se observou 2 c?es considerados curados clinicamente e 1 com melhora. J? nos grupos 4 e 5, evidenciou-se 4 e 5 animais curados, respectivamente e, tamb?m um com n?tida melhora do quadro cl?nico. O fluazuron 2,5% pour on n?o demonstrou efic?cia no tratamento da sarna demod?cica canina. J? a ivermectina 0,5% pour on associada ao fluazuron ou como terapia ?nica e, a ivermectina 3,15% por via subcut?nea nas duas dosagens diferentes, foram eficazes na redu??o do n?mero de ?caros ao final dos protocolos terap?uticos.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniv., 2006. / Härtill 5 uppsatser.

O cloridrato de tramadol é um analgésico de ação central, análogo sintético da codeína e morfina, o qual vem sendo amplamente estudado em equinos, sendo avaliado sua farmacocinética e farmacodinâmica. Em humanos, há relato de que o tramadol é substrato para a Gp-P, o que pode ser fator limitante na absorção do tramadol. A Gp-P funciona como uma bomba de efluxo celular, de maneira que, transporta ativamente xenobióticos do meio intracelular para o extracelular, atuando como um mecanismo de proteção contra xenobióticos. O estudo teve como objetivo a detecção do Gene MDR1 a partir do cDNA, e avaliar as alterações fisiológicas e efeito analgésico do tramadol em equinos submetidos a inibição da Gp-P entérica pela ivermectina. Seis equinos, machos e fêmeas, pesando 448±68Kg, foram distribuídos em três grupos autocontrole, recebendo tramadol por sonda nasogástrica na dose 1 mg/kg (GT1), 4 mg/kg (GT4) e recebendo tramadol 1mg/kg associada a ivermectina 0,2mg/kg VO. Foram avaliados FC, f, motilidade intestinal, temperatura corpórea aos 30 min antes, imediatamente antes da administração de qualquer substância para determinação dos valores basais e aos 30min, 60 min, 90 min, 120 min e a cada 60 min até 360 min após o tratamento. A claudicação foi avaliada aos 30 min, 60 min e a cada 60 min até os 360 min. Os parâmetros hemogasométricos foram avaliados no momento 0, 60 min e 120 min. Para as variáveis paramétricas utilizou-se análise de variância (ANOVA) para amostras pareadas, com posterior teste de Dunnett. Para comparações entre os grupos, realizou-se análise de variância, seguido de teste de Tukey. Para a variável não-paramétrica, motilidade intestinal, utilizou-se teste de Wilcoxon para amostras pareadas. As diferenças foram consideradas significantes quando P<0,05. Não foram observadas diferenças na FC e na avaliação analgésica. Houve hipomotilidade no GT1 e GT4 apenas ao final das avaliações e aumento da f em todos os grupos. Houve aumento do HCO3+ e redução do K+ e Ca++. Conclui-se que a inibição da Gp-P entérica pela ivermectina não alterou os efeitos do tramadol nas doses estudadas, sugerindo que o mesmo não é substrato para Gp-P, mas estudos futuros devem ser realizados a fim de avaliar a interação da ivermectina como inibidor da Gp-P na farmacocinética do tramadol. / Tramadol hydrochloride is a centrally acting analgesic, synthetic analogue of codeine and morphine, which has been widely studied in horses, being evaluated its pharmacokinetics and pharmacodynamics. In humans, there is a report that tramadol is a substrate for P-gp, which can be a limiting factor in the absorption of tramadol. P-gp acts as an efflux pump cell, that actively transports xenobiotics from the intracellular to the extracellular environment, acting as a protective mechanism against xenobiotic. The study aimed the detection of MDR1 Gene from cDNA, and the evaluation of physiological parameters and analgesic effect of tramadol in horses submitted to inhibition of enteric P-gp by ivermectin. Six horses, control of themselves, male and female, weighing 448 ± 68kg, were distributed into three groups, receiving tramadol by nasogastric tube in dose of 1 mg/kg (GT1), 4 mg/kg (GT4) and tramadol 1 mg/kg associated with ivermectin 0.2 mg/kg orally. Were evaluated HR, RR, intestinal motility, body temperature 30 min before, and immediately before the administration of any substance for determination of baseline and posterior at 30 min, 60 min, 90 min, 120 min and every 60 min up to 360 min after treatment. The analgesic evaluation occurred at 30 min, 60 min and every 60 min to 360 min. Blood gas parameters were evaluated at 0, 60 min and 120 min. For parametric variables were used analysis of variance (ANOVA) for paired samples, followed by Dunnett's test. For comparisons between groups, ANOVA followed by Tukey test were used. The non-parametric variable, intestinal motility, we used the Wilcoxon test for paired samples. Differences were considered significant when P <0.05. Differences in HR and analgesic evaluation were not observed. Hypomotility occurs in GT1 and GT4 only at the end of evaluation and RR increased in all groups. There was an increase of HCO3- and reduction of K+ and Ca++. We conclude that inhibition of enteric P-gp by ivermectin did not alter the effects of tramadol in the studied doses, suggesting that tramadol it is not a substrate for P-gp, but future studies should be conducted to assess the interaction of ivermectin as inhibitor of P-gp on the pharmacokinetics of tramadol.

O cloridrato de tramadol é um analgésico de ação central, análogo sintético da codeína e morfina, o qual vem sendo amplamente estudado em equinos, sendo avaliado sua farmacocinética e farmacodinâmica. Em humanos, há relato de que o tramadol é substrato para a Gp-P, o que pode ser fator limitante na absorção do tramadol. A Gp-P funciona como uma bomba de efluxo celular, de maneira que, transporta ativamente xenobióticos do meio intracelular para o extracelular, atuando como um mecanismo de proteção contra xenobióticos. O estudo teve como objetivo a detecção do Gene MDR1 a partir do cDNA, e avaliar as alterações fisiológicas e efeito analgésico do tramadol em equinos submetidos a inibição da Gp-P entérica pela ivermectina. Seis equinos, machos e fêmeas, pesando 448±68Kg, foram distribuídos em três grupos autocontrole, recebendo tramadol por sonda nasogástrica na dose 1 mg/kg (GT1), 4 mg/kg (GT4) e recebendo tramadol 1mg/kg associada a ivermectina 0,2mg/kg VO. Foram avaliados FC, f, motilidade intestinal, temperatura corpórea aos 30 min antes, imediatamente antes da administração de qualquer substância para determinação dos valores basais e aos 30min, 60 min, 90 min, 120 min e a cada 60 min até 360 min após o tratamento. A claudicação foi avaliada aos 30 min, 60 min e a cada 60 min até os 360 min. Os parâmetros hemogasométricos foram avaliados no momento 0, 60 min e 120 min. Para as variáveis paramétricas utilizou-se análise de variância (ANOVA) para amostras pareadas, com posterior teste de Dunnett. Para comparações entre os grupos, realizou-se análise de variância, seguido de teste de Tukey. Para a variável não-paramétrica, motilidade intestinal, utilizou-se teste de Wilcoxon para amostras pareadas. As diferenças foram consideradas significantes quando P<0,05. Não foram observadas diferenças na FC e na avaliação analgésica. Houve hipomotilidade no GT1 e GT4 apenas ao final das avaliações e aumento da f em todos os grupos. Houve aumento do HCO3+ e redução do K+ e Ca++. Conclui-se que a inibição da Gp-P entérica pela ivermectina não alterou os efeitos do tramadol nas doses estudadas, sugerindo que o mesmo não é substrato para Gp-P, mas estudos futuros devem ser realizados a fim de avaliar a interação da ivermectina como inibidor da Gp-P na farmacocinética do tramadol. / Tramadol hydrochloride is a centrally acting analgesic, synthetic analogue of codeine and morphine, which has been widely studied in horses, being evaluated its pharmacokinetics and pharmacodynamics. In humans, there is a report that tramadol is a substrate for P-gp, which can be a limiting factor in the absorption of tramadol. P-gp acts as an efflux pump cell, that actively transports xenobiotics from the intracellular to the extracellular environment, acting as a protective mechanism against xenobiotic. The study aimed the detection of MDR1 Gene from cDNA, and the evaluation of physiological parameters and analgesic effect of tramadol in horses submitted to inhibition of enteric P-gp by ivermectin. Six horses, control of themselves, male and female, weighing 448 ± 68kg, were distributed into three groups, receiving tramadol by nasogastric tube in dose of 1 mg/kg (GT1), 4 mg/kg (GT4) and tramadol 1 mg/kg associated with ivermectin 0.2 mg/kg orally. Were evaluated HR, RR, intestinal motility, body temperature 30 min before, and immediately before the administration of any substance for determination of baseline and posterior at 30 min, 60 min, 90 min, 120 min and every 60 min up to 360 min after treatment. The analgesic evaluation occurred at 30 min, 60 min and every 60 min to 360 min. Blood gas parameters were evaluated at 0, 60 min and 120 min. For parametric variables were used analysis of variance (ANOVA) for paired samples, followed by Dunnett's test. For comparisons between groups, ANOVA followed by Tukey test were used. The non-parametric variable, intestinal motility, we used the Wilcoxon test for paired samples. Differences were considered significant when P <0.05. Differences in HR and analgesic evaluation were not observed. Hypomotility occurs in GT1 and GT4 only at the end of evaluation and RR increased in all groups. There was an increase of HCO3- and reduction of K+ and Ca++. We conclude that inhibition of enteric P-gp by ivermectin did not alter the effects of tramadol in the studied doses, suggesting that tramadol it is not a substrate for P-gp, but future studies should be conducted to assess the interaction of ivermectin as inhibitor of P-gp on the pharmacokinetics of tramadol.

Submitted by MARCEL KENZO VILALBA ONIZUKA null (marcel.kenzo@gmail.com) on 2016-11-22T17:07:43Z No. of bitstreams: 1 Dissertação_Marcel_Kenzo_Vilalba_Onizuka.pdf: 966239 bytes, checksum: 3186b3d5035f1c543aa30c82f00aad7c (MD5) / Approved for entry into archive by Felipe Augusto Arakaki (arakaki@reitoria.unesp.br) on 2016-11-25T15:49:39Z (GMT) No. of bitstreams: 1 onizuka_mkv_me_jabo.pdf: 966239 bytes, checksum: 3186b3d5035f1c543aa30c82f00aad7c (MD5) / Made available in DSpace on 2016-11-25T15:49:39Z (GMT). No. of bitstreams: 1 onizuka_mkv_me_jabo.pdf: 966239 bytes, checksum: 3186b3d5035f1c543aa30c82f00aad7c (MD5) Previous issue date: 2016-10-20 / O objetivo deste trabalho foi avaliar o desenvolvimento ponderal de bovinos precoces, tratados estrategicamente contra nematódeos gastrintestinais, em julho e setembro, com dois fármacos distintos, o sulfóxido de albendazole 10% e ivermectina 1%. O experimento teve duração de 112 dias e foi dividido em três etapas: E1 (pasto); E2 (pasto e suplemento); E3 (confinamento e eutanásia). Foram utilizados 36 bovinos (Braford), idade entre 18 e 20 meses, peso médio inicial 357 kg e naturalmente infectados com nematódeos gastrintestinais. Os animais foram distribuídos em três grupos experimentais (n=12), de acordo com peso e OPG: GI (controle); GII (sulfóxido de albendazole 10%); GIII (ivermectina 1%). Para análise dos dados foram avaliadas as variáveis peso, ganho em peso, OPG, helmintos remanescentes e rendimento de carcaça. Ao término do período experimental os animais foram eutanasiados para realizar a necropsia parasitológica. Em todas as análises estabeleceu-se como nível de significância 5%. Houve diferença significativa nas variáveis peso, ganho em peso, OPG e rendimento de carcaça quando comparou-se o GII aos demais grupos. Na necropsia parasitológica foi encontrada predominância das espécies Trichostrongylus axei e Cooperia punctata. Concluiu-se que os bovinos tratados estrategicamente com sulfóxido de albendazole 10%, em julho e setembro, apresentaram melhor desenvolvimento ponderal em relação aos demais grupos. / The objective of this study was to evaluate the weight development of early cattle strategically treated against gastrointestinal nematodes in July and September, with two different drugs, albendazole sulphoxide 10% and ivermectin 1%. The experiment lasted 112 days and was divided into three stages: S1 (pasture); S2 (pasture and supplement); S3 (feedlot and euthanasia). Were used 36 cattle (Braford), aged between 18 and 20 months, average weight 357 kg and naturally infected with gastrointestinal nematodes. The animals were divided into three experimental groups (n= 12), according to weight and EPG: GI (control); GII (albendazole sulphoxide 10%); GIII (ivermectin 1%). For data analysis was evaluated the variables weight, weight gain, EPG, remaining helminths and carcass yield. At the end of the experimental period the animals were euthanized to perform the parasitological necropsy. In all analyzes it was established as a significance level of 5%. There were significant differences in the variables weight, weight gain, EPG and carcass yield when compared to the GII to other groups. In parasitological necropsy found predominance of species Trichostrongylus axei and Cooperia punctata. It was concluded that cattle strategically treated with albendazole sulphoxide 10%, in July and September, showed better growth development than the other groups.

Orientador: Alvimar José da Costa / Resumo: O objetivo deste trabalho foi avaliar o desenvolvimento ponderal de bovinos precoces, tratados estrategicamente contra nematódeos gastrintestinais, em julho e setembro, com dois fármacos distintos, o sulfóxido de albendazole 10% e ivermectina 1%. O experimento teve duração de 112 dias e foi dividido em três etapas: E1 (pasto); E2 (pasto e suplemento); E3 (confinamento e eutanásia). Foram utilizados 36 bovinos (Braford), idade entre 18 e 20 meses, peso médio inicial 357 kg e naturalmente infectados com nematódeos gastrintestinais. Os animais foram distribuídos em três grupos experimentais (n=12), de acordo com peso e OPG: GI (controle); GII (sulfóxido de albendazole 10%); GIII (ivermectina 1%). Para análise dos dados foram avaliadas as variáveis peso, ganho em peso, OPG, helmintos remanescentes e rendimento de carcaça. Ao término do período experimental os animais foram eutanasiados para realizar a necropsia parasitológica. Em todas as análises estabeleceu-se como nível de significância 5%. Houve diferença significativa nas variáveis peso, ganho em peso, OPG e rendimento de carcaça quando comparou-se o GII aos demais grupos. Na necropsia parasitológica foi encontrada predominância das espécies Trichostrongylus axei e Cooperia punctata. Concluiu-se que os bovinos tratados estrategicamente com sulfóxido de albendazole 10%, em julho e setembro, apresentaram melhor desenvolvimento ponderal em relação aos demais grupos. / Mestre