Orientador: José Antonio Marques / Coorientador: Sérgio Britto Garcia / Banca: Deborah Penteado Martins Dias / Banca: Paulo Aléscio Canola / Resumo: A Dermatite Digital Bovina (DDB) constitui uma das principais causas de graus elevados de claudicação em bovinos leiteiros, em função da dor que estes animais aparentam frente ao estímulo nocioceptivo. A hiperalgesia em alguns casos de dor crônica está relacionada à expressão exacerbada de fibras dos receptores vaniloides TRPV1, podendo haver forte correlação com casos crônicos da DDB. No presente estudo foram utilizados 15 bovinos, fêmeas, da raça Holandesa Preto e Branco, com idades de 2 a 7 anos, em lactação com pico médio de 47,85 L, mantidos em regime "free-stall". Durante o casqueamento realizado na propriedade, as lesões da DDB foram identificadas e divididas em quatro grupos, referentes aos quatro estágios da doença (inicial M1, clássico M2, intermediário M3 e crônico M4). Foram coletadas biópsias por meio de "punch" cutâneo (4mm). Além das lesões foram coletadas amostras de pele sadia de cada animal. Priori às biopsias realizou-se a dimensionamento das lesões, para média comparativa entre os estágios. Os fatores predisponentes ao aparecimento de lesões (idade, número de partos e pico de lactação) foram considerados. As biopsias foram processadas em laboratório e colocadas frente à reação imunológica com anticorpos anti-TRPV1 (Chemicon -USA). Posteriormente as fibras imuno-marcadas nos quatro grupos e pele sadia foram contabilizadas e comparadas. As dimensões das lesões foram analisadas pelo método estatístico descritivo e possuíam média de comprimento e largura no es... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The Bovine Digital Dermatitis (BDD) is one of the main cause on high claudication levels at the dairy cattle, as a result of the pain that appear in those animal front of the nociceptive stimulus. Hyperalgesia in some chronic pain cases is related to the exacerbated expression of vanyloid TRPV1 fibers, and may have a hole on the chronic cases of BDD. In the present study, 15 Black and White Holstein cow was used, aged 2 to 7 years old, during lactation with mean peak of 47.85L, kept in the free-stall under the same diet. BDD lesions were identified and divided into 4 stages of the disease (initial M1, classic M2, intermediate M3 and chronic4), which skin are collected by dermal punch (4mm). In addition to the lesions, healthy skin samples were collected from each animal. Prior to the biopsies, the lesion dimensions were taken, for comparative average between the stages. Predisposing factors to the lesions appearance (age, number of births and lactation peak) were considered. The biopsies were processed at the laboratory and reacted with anti-TRPV1 antibodies (Chemicon -USA). The immuno-marked fibers in the four groups and healthy skin were counted and compared. The lesion dimensions were analyzed by descriptive statistical method and had a M1 mean length/ width on M1 of 5,60mm±3,20 x 4,4±1,34mm, M2 of 12,60±6,46mm x 14,4±8,87mm, M3 of 21,60±3,36mm x 17,20±6,61mm and M4 of 21,60±3,36mm x 24,57±7,32mm. The thickness mean value in M1 of 1.80 ± 1.09mm, M2 of 6.20 ± 2.16mm, M3 of 7.40 ± 6.54mm and M4 of 8.85 ± 4.14mm. The predisposing factors results were analyzed separately with the number of lesions by Pearson Correlation statistic method (p <0.05). There was difference between more lesion and birth numbers. TPV1 fiber counts were analyzed by logarithmic scale (p <0.05), with a significant difference between the M4 group (chronic stage of DDB) and the other groups. / Mestre

Orientador: Celso Antonio Rodrigues / Banca: José Luiz de Mello Nicoletti / Banca: José Wanderley Cattelan / Resumo: A Dermatite Digital Papilomatosa (DDP) é uma enfermidade com etiologia bacteriana associadas às espiroquetas. O tratamento consiste na administração de antimicrobianos parenterais ou tópicos, destacando-se especialmente as tetraciclinas. Os objetivos deste trabalho foram analisar e comparar a eficiência do tratamento tópico e sistêmico com oxitetraciclina em vacas acometidas por DDP, e a presença de resíduos deste antimicrobiano no leite e suas concentrações no líquido sinovial e plasma. Foram utilizadas 8 vacas no grupo 1 tratadas topicamente e 8 no grupo 2 tratadas por via sistêmica . As amostras foram coletadas nos momentos M0: anterior aos tratamentos, M1: 6 horas pós-tratamento, M2: 12 horas após, e em intervalos de 12 horas até o M23. A eficiência dos tratamentos foi avaliada através do grau de claudicação e da extensão da lesão. As concentrações do antimicrobiano de oxitetraciclina foram determinadas por HPLC. As vacas do Grupo 1 apresentaram redução das lesões e no grau de claudicação e as do grupo 2 não demonstraram melhora nas lesões e elevação no grau de claudicação. As concentrações avaliadas em leite, líquido sinovial e plasma no Grupo 1, revelaram valores abaixo dos limites de detecção estabelecidos pelo HPLC. A avaliação dos resíduos de oxitetraciclina no leite do Grupo 2, entre M1 e M23, encontram-se acima dos limites máximos estabelecidos pela legislação. Conclui-se que a oxitetraciclina em pó foi eficiente para o tratamento de DDP e não apresenta resíduos em plasma, líquido sinovial e leite e que o tratamento sistêmico apresenta baixa eficiência e elevadas concentrações no leite / Abstract: Papillomatous digital dermatitis (PDD) is a restricted digital disease caused by a spirochete-like and other bacterias. The treatment consists of administrating systemic or topics antibiotics, especially tetracyclines. With this work it is intended to compare the efficiency of the topical and systemic treatment with oxitetracycline in cows with PDD and analyze the presence of residues of this antibiotic in the milk and its concentration in the sinovial liquid and plasma. The total of 16 lactating cows with PDD were divided in 2 treatment groups. The samples were obtained in the moments M0: previous to the treatments, M1: 6 hours pos treatment, M2: 12 hours after treatment, and in intervals of 12 hours after treatment until M23. The efficiency of the treatment was evaluated through the lameness degree and the extension of the lesion. The assayed for oxitetracycline content was measured by means of HPLC. The cows of the Group 1 presented reduction of the lesions and in the lameness degree. Cattle of the Group 2 did not demonstrate improvement in the lesions and showed elevation in the lameness degree. The concentration of sinovial fluid and plasma in the milk of cows belonging to Group 1 revealed values below the established detection limits for HPLC in milk. The evaluation of the residues of oxitetracycline in the milk of the Group 2, between M1 and M23, were above the maximum limits established by legislation. As a matter of conclusion it is argued that the powdered oxitetracycline was efficient for the treatment of PDD and it does not present residues in milk, level of plasma or sinovial fluid. Otherwise, the systemic treatment presents low efficiency and high concentration of such substance in the milk / Mestre

Digital dermatitis is a top cause of lameness in dairy cattle that results in ulcerative lesions on the feet. Topical salicylic acid has been shown to provide similar efficacy to the antibiotic drugs used previously, but there is no milk withholding time established in the United States. The objective of this study was to provide data in order to establish this withholding period. A secondary objective was to evaluate outcomes among treatments. Treatment groups were topical applications of the following drugs: salicylic acid paste, salicylic acid powder, and tetracycline. The lesions were scored at day 0, day 7, and day 28 post-treatment. Milk samples were collected the day before treatment, 4 hours, 8 hours, 24 hours, 36 hours, and 48 hours post-treatment. Results indicated that most cows did not show detectable levels of salicylic acid after 24 hours.

A prospective study was performed to evaluate a biological medication in the treatment of digital dermatitis (DD) in dairy cattle. The study was divided into four parts; i) on farm evaluation of DD and treatment effects and comparison between the biological ointment and OTC-spray, ii) statistical evaluation, iii) histological examination using FISH and iv) microbiological examination and culture if bacteria found in biopsies from infected skin.

Made available in DSpace on 2015-06-17T19:34:49Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-07-22. Added 1 bitstream(s) on 2015-06-18T12:47:00Z : No. of bitstreams: 1 000831292.pdf: 3855958 bytes, checksum: f0eddfd2bd8c33b972b6c00be59dda5a (MD5) / O objetivo do estudo foi testar um protocolo de tratamento para pododermatite classe III em um grupo de Spheniscus magellanicus mantidos em cativeiro utilizando terapia fotodinâmica (TFD) e compará-lo com outro tratado com antibióticos (ATB). Todos os animais foram submetidos inicialmente à debridamento cirúrgico da área da lesão e em seguida aleatoriamente separados em um grupo TFD e outro ATB, contendo 11 lesões cada. No grupo ATB utilizou-se pomada antibiótica três vezes por semana e antibiótico e anti-inflamatório sistêmicos diariamente. No grupo TFD, terapia fotodinâmica foi aplicada três vezes por semana sem a utilização de medicação sistêmica. Fotografias da área da lesão, swab para cultura e antibiograma e sangue para hemograma e bioquímica sanguínea foram coletados no início e a cada 14 dias perfazendo um total de 84 dias. Os resultados reafirmaram a natureza recidivante dessas lesões que se iniciam a partir do ponto de apoio. A prevalência das bactérias ao longo do tempo não foi uniforme, sendo a realização de antibiogramas para escolha de uma droga apropriada necessário em diferentes momentos. Na comparação, o grupo TFD apresentou desempenho melhor com 63,62% de cura em um tempo médio de 42 dias, contra 9,09% de cura em um tempo médio de 70 dias no grupo ATB. Os achados do presente estudo atestam a eficácia da Terapia Fotodinâmica para o tratamento de pododermatite classe III para Spheniscus magellanicus / The aim of the study was to test a treatment protocol for pododermatitis class III in a group of captive Spheniscus magellanicus using photodynamic therapy (PDT) and compare it with another treated with antibiotics (ATB). All animals initially underwent surgical debridement of the lesion area and then they were randomly divided into one TFD and one ATB group, each containing 11 injuries. In the ATB group antibiotic ointment was used three times a week and systemic antibiotic and anti-inflammatory daily. In PDT group, photodynamic therapy has been applied three times a week without the use of systemic medication. Photos of the lesion area, swab for culture and sensitivity and blood for blood count and blood chemistry were collected on the first day and every 14 days for a total of 84 days. The results reaffirmed the relapsing nature of these lesions that begin from the point of support. The prevalence of bacteria over time was not uniform and there was need to carry out antibiograms at different times to select an appropriate drug. In comparison, the PDT group showed the best performance with 63.62% on an average healing time of 42 days, compared to 9.09% in an average healing time of 70 days in the ATB group. The findings of this study attest to the effectiveness of photodynamic therapy for the treatment of class III to pododermatitis Spheniscus magellanicus

Orientador: Carlos Roberto Teixeira / Banca: Sheila Canevese Rahal / Banca: Alyne Simões Gonçalves / Resumo: O objetivo do estudo foi testar um protocolo de tratamento para pododermatite classe III em um grupo de Spheniscus magellanicus mantidos em cativeiro utilizando terapia fotodinâmica (TFD) e compará-lo com outro tratado com antibióticos (ATB). Todos os animais foram submetidos inicialmente à debridamento cirúrgico da área da lesão e em seguida aleatoriamente separados em um grupo TFD e outro ATB, contendo 11 lesões cada. No grupo ATB utilizou-se pomada antibiótica três vezes por semana e antibiótico e anti-inflamatório sistêmicos diariamente. No grupo TFD, terapia fotodinâmica foi aplicada três vezes por semana sem a utilização de medicação sistêmica. Fotografias da área da lesão, swab para cultura e antibiograma e sangue para hemograma e bioquímica sanguínea foram coletados no início e a cada 14 dias perfazendo um total de 84 dias. Os resultados reafirmaram a natureza recidivante dessas lesões que se iniciam a partir do ponto de apoio. A prevalência das bactérias ao longo do tempo não foi uniforme, sendo a realização de antibiogramas para escolha de uma droga apropriada necessário em diferentes momentos. Na comparação, o grupo TFD apresentou desempenho melhor com 63,62% de cura em um tempo médio de 42 dias, contra 9,09% de cura em um tempo médio de 70 dias no grupo ATB. Os achados do presente estudo atestam a eficácia da Terapia Fotodinâmica para o tratamento de pododermatite classe III para Spheniscus magellanicus / Abstract: The aim of the study was to test a treatment protocol for pododermatitis class III in a group of captive Spheniscus magellanicus using photodynamic therapy (PDT) and compare it with another treated with antibiotics (ATB). All animals initially underwent surgical debridement of the lesion area and then they were randomly divided into one TFD and one ATB group, each containing 11 injuries. In the ATB group antibiotic ointment was used three times a week and systemic antibiotic and anti-inflammatory daily. In PDT group, photodynamic therapy has been applied three times a week without the use of systemic medication. Photos of the lesion area, swab for culture and sensitivity and blood for blood count and blood chemistry were collected on the first day and every 14 days for a total of 84 days. The results reaffirmed the relapsing nature of these lesions that begin from the point of support. The prevalence of bacteria over time was not uniform and there was need to carry out antibiograms at different times to select an appropriate drug. In comparison, the PDT group showed the best performance with 63.62% on an average healing time of 42 days, compared to 9.09% in an average healing time of 70 days in the ATB group. The findings of this study attest to the effectiveness of photodynamic therapy for the treatment of class III to pododermatitis Spheniscus magellanicus / Mestre

A prospective study was performed to evaluate a biological medication in the treatment of digital dermatitis (DD) in dairy cattle. The study was divided into four parts; i) on farm evaluation of DD and treatment effects and comparison between the biological ointment and OTC-spray, ii) statistical evaluation, iii) histological examination using FISH and iv) microbiological examination and culture if bacteria found in biopsies from infected skin.

Made available in DSpace on 2014-07-29T15:13:42Z (GMT). No. of bitstreams: 1 tese pos defesa.pdf: 2322225 bytes, checksum: 4027ee4b5a7e38a6c0d20eb192a18551 (MD5) Previous issue date: 2011-03-18 / This study has aimed do evaluate a risk factor for bovine digital dermatitis and the efficacy of 5% aqueous barbatimão extract solution as ancillary in interdigital skin incisional wound repair. One hundred bovine females, 50 Holstein and 50 Girolando, from two different farms were used for the risk factor study. Experimental units were divided into four groups: healthy Holstein cattle group (GHS) and digital dermatitis Holstein cattle group (GHL), healthy Girolando cattle group (GGS) and digital dermatitis Girolando cattle group (GGL). Ten milliliters of blood were collected from all animals and stored in anticoagulant added tubes. Samples were refrigerated and submitted to hemolytic tests for eight bovine blood systems: A, B, C, F, J, L, S and Z. The frequency of blood factors was determined and compared, in each system and each group. Data were analyzed through non parametric tests: Chi Square and Fisher s exact. The results indicated that the blood factors may contribute to the occurrence of digital dermatitis lesions, but further studies are needed for a more accurate conclusion. Thirty confined, 10 months old calves were selected in order to evaluate the efficacy of 5% barbatimão solution. A biopsy punch was introduced in each interdigital space, up to the dermis, to produce a lesion measuring one centimeter in diameter. The lesions were numbered 120 in total and named experimental units, which were divided into two groups of 60 units each: GB treated with 5% aqueous barbatimão bark extract solution and GC treated with water. Tissue repair was evaluated through macroscopic and histological findings in days three, seven, 14 and 21 of treatment. Macroscopic findings were analyzed descriptively, while histological findings were submitted to Mann Whitney test, with p<0.05. For lesions treated, GB group showed less intense polymorphonuclear cells infiltration on the first 14 days of treatment. A marked collagen deposition was also observed at day 14 while collagen alignment was more expressive on day 21. More specific studies may be necessary in order to determine if blood factors can contribute to the occurrence of digital dermatitis. Barbatiman 5% footbath solution for confined cattle helps qualitatively in modulation of interdigital cutaneous incisional wound repair, but does not avoid the extension of inflammatory phase of healing. / Objetivou-se nesta pesquisa estudar um fator de risco para a dermatite digital bovina e avaliar a eficácia do extrato de barbatimão em solução aquosa a 5% como auxiliar no reparo de feridas cutâneas induzidas no espaço interdigital de bovinos. Para pesquisar o fator de risco utilizaram-se 100 fêmeas bovinas, sendo 50 Holandesas e 50 Girolando, de duas propriedades rurais. Dividiram-se as unidades experimentais em quatro grupos: grupo de animais saudáveis da raça Holandesa (GHS) e grupo com lesões de dermatite digital (GHL); grupo de animais saudáveis da raça Girolando (GGS) e grupo com lesões de dermatite digital (GGL). De todos os animais colheram-se 10 ml de sangue, em tubo a vácuo, contendo anticoagulante. As amostras foram refrigeradas e submetidas a testes hemolíticos, para oito sistemas sanguíneos de bovinos: A, B, C, F, J, L, S e Z. A frequência dos fatores sanguíneos foi determinada e comparada, dentro de cada sistema, em cada grupo de animais. Os dados foram analisados através dos testes não paramétricos de Qui-quadrado e Exato de Fisher. Pelos resultados obtidos concluiu-se que os fatores sanguíneos podem contribuir para a ocorrência de lesões de dermatite digital, contudo são necessários estudos mais específicos para uma conclusão mais precisa. Para avaliar a eficácia da solução de barbatimão a 5%, utilizaram-se 30 animais com dez meses de idade, mantidos em sistema de confinamento. Em cada espaço interdigital dos animais, com auxílio de um punch de um centímetro de diâmetro, produziu-se uma lesão cutânea até a derme, totalizando 120 feridas. As feridas foram denominadas unidades experimentais e distribuídas em dois grupos com 60 unidades cada: grupo GB tratado com solução de extrato de barbatimão a 5% e grupo GC tratado com água. A evolução do processo de reparação tecidual das feridas foi acompanhada por meio de avaliações macroscópicas e histológicas nos dias três, sete, 14 e 21 de tratamento. Os achados macroscópicos foram analisados descritivamente e os histológicos submetidos ao teste de Mann Whitney, com p<0,05. Para as feridas tratadas, observou-se que o grupo GB apresentou menor intensidade de infiltrado de células polimorfonucleares nos primeiros 14 dias de tratamento. Nesse grupo observou-se ainda que a deposição de colágeno mostrou-se mais acentuada aos 14 dias e aos 21 dias as fibras colágenas apresentavam-se mais alinhadas. O uso do extrato de barbatimão a 5% em pedilúvio, em bovinos confinados, auxiliou qualitativamente no processo de modulação da reparação de feridas cutâneas interdigitais incisionais, mas não impediu o prolongamento da fase inflamatória do processo cicatricial.

Bovine Dermatitis digitalis (DD) ist eine weltweit verbreitete Infektionskrankheit bei Rindern, die primär die plantare Haut über dem Kronenrand nahe des Zwischenzehenspalts der Hinterklauen betrifft. Schmerzhafte ulzero-proliferative Läsionen mit akuten und chronischen Erscheinungsformen führen zu Verhaltensänderungen und Lahmheit der Tiere. DD hat damit einen erheblichen Einfluss auf deren Wohl und ihre Leistungen. Zahlreiche Untersuchungen zur Ätiologie der Krankheit ergaben, dass es sich um das Zusammenspiel verschiedener Ursachen handelt. Einer synergistischen multifaktoriellen Infektion mit starker Beteiligung von Bakterien der Gattung Treponema kommt dabei besondere Bedeutung zu. Aspekte wie Tierhaltung, Hygienestandards und genetische Prädispositionen wurden ebenfalls intensiv untersucht. Nichtsdestotrotz bleiben Infektionsherde, Transmissionsrouten und Pathomechanismen weitgehend unklar. Zum besseren Verständnis der Ereignisse, die zu DD-Läsionen führen, sollte im Zuge dieser Arbeit ein organotypisches Zellkulturmodell der bovinen Haut erstellt werden, welches in späteren Versuchen mit dem Krankheitserreger zum Einsatz kommen soll. Verlässliche und reproduzierbare Techniken zur Isolation und Kultur von bovinen primären Keratinozyten und Fibroblasten wurden etabliert; geeignete Zellkulturmedien für die Langzeitkultivierung und –aufbewahrung der Hautzellen wurden identifiziert. Zur Erstellung des Hautmodells wurden zwei verschiedene Ansätze miteinander verglichen. Der zweite Ansatz, bei dem Keratinozyten direkt auf ein dermales Äquivalent, d.h. ein Pad aus bovinem Kollagen I mit eingesäten post-mitotischen Fibroblasten, gesät wurden, brachte ein vielversprechendes Hautmodell hervor. Die inkorporierten post-mitotischen Fibroblasten wiesen eine charakteristische Zellmorphologie mit intakten Nuklei auf. Die terminale Differenzierung der Keratinozyten auf dem dermalen Äquivalent wurde mittels Immunfluoreszenzfärbungen mit Antikörpern gegen die Markerproteine Keratin 14 und Desmoglein 1 gezeigt. Die Ergebnisse erster Experimente mit Treponema spp. verdeutlichen, dass das Hautäquivalent ein geeignetes Modell zur Untersuchung der Pathogenese der DD darstellt. / Bovine digital dermatitis (DD) is a worldwide occurring, infectious disease in cattle primarily affecting the plantar skin above the coronary band near the interdigital cleft on hind feet. Painful ulceroproliferative lesions with acute and chronic appearances lead to behavioral changes and lameness. Hence, DD has a major impact on animal welfare and performance. Substantial efforts in investigating the etiology of the disease revealed a synergistic origin with evidence for a multibacterial infection and the strong involvement of bacteria from the genus Treponema. As the interaction between host, pathogen and environment is not negligible, surrounding circumstances such as housing, general hygiene and genetic predispositions have been investigated intensively. Nevertheless, infection reservoirs, transmission routes and pathomechanisms remain widely unclear. To better understand the cellular and molecular events during Treponema-infection of bovine skin, it was the specific aim of this study to establish an organotypic in vitro skin model, which could be challenged with the causative agent of the disease. A technique to reliably and reproducibly isolate primary keratinocytes and fibroblasts from the site of infection was established. Appropriate cell culture media for the long-term cultivation and storage of bovine skin cells were identified. Two different methods to develop the skin model were compared. The second strategy in which keratinocytes were directly seeded on top of a dermal equivalent, i.e. a bovine collagen type I pad with embedded post-mitotic fibroblasts, gave rise to a promising organotypic skin equivalent. The incorporated post-mitotic fibroblasts showed a characteristic cell morphology with intact nuclei. The terminal differentiation of the keratinocytes on top of the dermal equivalent was shown with anti-K14 and anti-Dsg1 immunofluorescence stainings. The results of initial Treponema-experiments proved that the skin equivalent is a suitable model to investigate the underlying mechanisms during Treponema-infection of bovine skin and hence, the pathogenesis of DD.