Tesis sobre el tema "Trichostrongylus colubriformis"

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Com o objetivo de se investigar a resposta imunológica e fisiopatologia das infecções por Trichostrongylus colubriformis em cordeiros Santa Inês, trinta ovinos foram agrupados em 10 classes, de acordo com seu peso vivo, sendo cada uma destas constituída de três animais. Por sorteio, um animal de cada classe constituiu o grupo infectado artificialmente com 2500 larvas de T. colubriformis, três vezes por semana, ao longo de 13 semanas e alimentado ad libitum, outro animal constituiu o grupo não infectado, alimentado com a mesma quantidade consumida pelo animal infectado, de mesma classe, no dia anterior, e o último animal constituiu o grupo não infectado e alimentado ad libitum. Diariamente as sobras de alimento eram pesadas, para que se estimasse o consumo alimentar individual de cada cordeiro. Semanalmente os animais eram pesados e amostras de sangue e fezes eram colhidas para a realização de exames hematológicos, imunológicos e coproparasitológicos. Ao final deste estudo, os cordeiros foram eutanaziados para a determinação da carga parasitária e colheita de amostras de tecido e muco intestinal para realização de análises histopatológicas e imunológicas. Os resultados demonstraram que a resposta imunológica impediu parcialmente o estabelecimento das larvas infectantes nos cordeiros infectados, porém houve redução do desempenho produtivo nos animais, independentemente da carga parasitária. O grupo infectado apresentou a pior conversão alimentar (88,81 ± 21,19 g a cada 1000g de alimento consumido), sendo esta significativamente menor (P<0,01) aos outros dois grupos experimentais. Os efeitos deletérios nos cordeiros foram decorrentes tanto das alterações patológicas causadas pelos vermes adultos na mucosa intestinal, como das alterações imunopatológicas produzidas pelo contato com as larvas infectantes. A alta demanda de nutrientes destinados ao desenvolvimento...
To investigate the immune response and physiopathology of Trichostrongylus colubriformis infections in Santa Inês lambs, thirty sheep were grouped into 10 classes of three animals each according to their liveweight. From each class, one animal was randomly selected to constitute the group artificially infected with 2500 T. colubriformis larvae three times every week for 13 weeks and fed ad libitum; another animal included the non-infected group fed with the same quantity consumed by the infected animal of the same class on the previous day; and one third animal constituted the non-infected group fed ad libitum. Leftovers were daily weighed to assess food consumption for each lamb. Every week, the animals were weighed and blood and fecal samples were collected for hematological, immunological and coproparasitological tests. At the end of this study, lambs were euthanized to detect worm burden and collect intestinal mucus and tissue samples for histopathological and immunological analyses. The immune response partially prevented the establishment of infective larvae in infected lambs. However, there was a reduction in the productive performance of these animals independently of their worm burden. The Infected Group had the worst feed conversion (88.81 ± 21.19 g in body weight for each 1000 g food consumed), this it was significantly lower (P<0.01) than that of the other two experimental groups. Deleterious effect in the lambs were due to pathological changes caused by adult nematodes in the intestinal mucosa and to immunopathological changes resultant of the contact with infective larvae. The high nutrient demand for the development and maintenance of the immune response and the repair of damaged tissues during infections also contributes in the low performance presented infected animals.

Orientador: Alessandro Francisco Talamini do Amarante
Banca: Maria Cecilia Rui Luvizotto
Banca: Rogerio Martins Amorin
Resumo: Com o objetivo de se investigar a resposta imunológica e fisiopatologia das infecções por Trichostrongylus colubriformis em cordeiros Santa Inês, trinta ovinos foram agrupados em 10 classes, de acordo com seu peso vivo, sendo cada uma destas constituída de três animais. Por sorteio, um animal de cada classe constituiu o grupo infectado artificialmente com 2500 larvas de T. colubriformis, três vezes por semana, ao longo de 13 semanas e alimentado ad libitum, outro animal constituiu o grupo não infectado, alimentado com a mesma quantidade consumida pelo animal infectado, de mesma classe, no dia anterior, e o último animal constituiu o grupo não infectado e alimentado ad libitum. Diariamente as sobras de alimento eram pesadas, para que se estimasse o consumo alimentar individual de cada cordeiro. Semanalmente os animais eram pesados e amostras de sangue e fezes eram colhidas para a realização de exames hematológicos, imunológicos e coproparasitológicos. Ao final deste estudo, os cordeiros foram eutanaziados para a determinação da carga parasitária e colheita de amostras de tecido e muco intestinal para realização de análises histopatológicas e imunológicas. Os resultados demonstraram que a resposta imunológica impediu parcialmente o estabelecimento das larvas infectantes nos cordeiros infectados, porém houve redução do desempenho produtivo nos animais, independentemente da carga parasitária. O grupo infectado apresentou a pior conversão alimentar (88,81 ± 21,19 g a cada 1000g de alimento consumido), sendo esta significativamente menor (P<0,01) aos outros dois grupos experimentais. Os efeitos deletérios nos cordeiros foram decorrentes tanto das alterações patológicas causadas pelos vermes adultos na mucosa intestinal, como das alterações imunopatológicas produzidas pelo contato com as larvas infectantes. A alta demanda de nutrientes destinados ao desenvolvimento... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: To investigate the immune response and physiopathology of Trichostrongylus colubriformis infections in Santa Inês lambs, thirty sheep were grouped into 10 classes of three animals each according to their liveweight. From each class, one animal was randomly selected to constitute the group artificially infected with 2500 T. colubriformis larvae three times every week for 13 weeks and fed ad libitum; another animal included the non-infected group fed with the same quantity consumed by the infected animal of the same class on the previous day; and one third animal constituted the non-infected group fed ad libitum. Leftovers were daily weighed to assess food consumption for each lamb. Every week, the animals were weighed and blood and fecal samples were collected for hematological, immunological and coproparasitological tests. At the end of this study, lambs were euthanized to detect worm burden and collect intestinal mucus and tissue samples for histopathological and immunological analyses. The immune response partially prevented the establishment of infective larvae in infected lambs. However, there was a reduction in the productive performance of these animals independently of their worm burden. The Infected Group had the worst feed conversion (88.81 ± 21.19 g in body weight for each 1000 g food consumed), this it was significantly lower (P<0.01) than that of the other two experimental groups. Deleterious effect in the lambs were due to pathological changes caused by adult nematodes in the intestinal mucosa and to immunopathological changes resultant of the contact with infective larvae. The high nutrient demand for the development and maintenance of the immune response and the repair of damaged tissues during infections also contributes in the low performance presented infected animals.
Mestre

Objetivando-se avaliar variáveis hemato-bioquímicas, digestibilidade aparente dos nutrientes e desempenho animal, assim como, o metabolismo e a cinética do fósforo (P) em cordeiros infectados experimentalmente com Trichostrongylus colubriformis, foram realizados dois experimentos. No primeiro estudo, foram utilizados 18 cordeiros machos, da raça Santa Inês, distribuídos em dois tratamentos experimentais: grupo infectado (I, n=9) e grupo controle (C, n=9) sem infecção. Os cordeiros do grupo I receberam aproximadamente 5000 L3, três vezes por semana, durante três semanas, totalizando 45000 L3 de T. colubriformis. O controle do consumo do alimento foi diário, utilizando a metodologia do tipo pair-fed (para eliminar efeito de consumo). Foram avaliados quinzenalmente, durante 75 dias, os dados de peso vivo, escore de condição corporal e amostras de sangue e fezes foram colhidas para determinação do hemograma, bioquímica sérica e contagem de ovos por grama de fezes (OPG), respectivamente. Também foi avaliado a digestibilidade aparente dos nutrientes e os parâmetros ruminais dos cordeiros. Os cordeiros apresentaram infecção moderada (OPG = 620,3 ± 594,98) com redução de eritrócitos, hemoglobina, hematócrito, volume corpuscular médio, concentração de hemoglobina corpuscular média e proteínas totais (P < 0,1). Por outro lado, houve aumento do número de plaquetas, bem como a quantidade de eosinófilos circulantes quando comparado aos cordeiros do grupo C (P < 0,1). Não houve alteração no consumo voluntário dos cordeiros I (P > 0,05), porém houve menor digestibilidade da MS (P < 0,1). Diante disso, conclui-se que a infecção por T. colubriformis alterou parâmetros hematológicos, bioquímicos, não interferindo no consumo de matéria seca, mas, reduzindo digestibilidade da MS em cordeiros infectados. No segundo estudo, objetivou-se avaliar o metabolismo e a cinética do P em cordeiros infectados experimentalmente com T. colubriformis utilizando a técnica de diluição isotópica e modelagem. Para tanto, foram utilizados 15 cordeiros distribuídos nos tratamentos infectado (I, n=8) e controle (C, n=7) como descrito acima. Após 66 dias da última inoculação foi injetado 6,6 MBq de 32P/cordeiro para avaliar a cinética do mineral. Amostras de sangue, fezes e urina foram coletados nos sete dias seguintes, com abate dos cordeiros no último dia para colheita de amostras dos tecidos ósseo e moles (fígado, rim, coração e músculo). Para análises dos fluxos de P foi utilizado o modelo biomatemático descrito por Lopes et al. (2001). Foi verificado consumo semelhante de P (VI) entre os cordeiros dos dois tratamentos (P > 0,1), menor absorção das frações do P endógeno (Vaf) e dietético (Vaa) bem como maior excreção do P de origem dietético (VFD) nos cordeiros I (P < 0,1). Com a menor absorção (VaT) de P em cordeiros I, houve, consequentemente, menor distribuição e menor incorporação de P nos ossos (VO+D). Conclui-se que o metabolismo de P de cordeiros infectados por T. colubriformis foi alterado, com redução da sua absorção intestinal e biodisponibilidade, com aumento da perda fecal e redução do fluxo de P para ossos
Aiming to evaluate hemato-biochemical variables, apparent digestibility and performance as well as the metabolism and kinetic of phosphorus (P) in lambs experimentally infected with Trichostrongylus colubriformis, two experiments were conducted. At the first study, 18 male lambs were distributed in two experimental treatments: infected group (I, n=9) and control group (C, n=9) unaffected. Infected lambs received approximately 5000 T. colubriformis infective larvae (L3), three times per week, during three weeks, totalling 45 000 T. colubriformis L3. The control of daily feed intake was performed (pair-fed). Data of live weight, body condition score and samples of blood and faeces were collected fortnightly, during 75 days to analyze the hemogram, biochemical variables and faecal egg count (FEC), respectively. In addition, apparent digestibility and ruminal parameters of lambs were determined. Lambs presented moderate infection (FEC = 620.3 ± 594.98) with decreased erythrocytes, haemoglobin, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin concentration and total protein. On the other hand, increased platelets counts as well as the amount of circulating eosinophils when compared to the group C (P < 0.1). The voluntary intake of infected lambs was not altered, but there was lower dry matter digestibility. It was concluded that T. colubriformis infection altered haematological and biochemical parameters. It did not interfere in dry matter intake, but decreased digestibility in infected lambs. In the second study, the objective was to study the metabolism and kinetic of P in lambs experimentally infected with T. colubriformis using isotope dilution technique and modelling. For this, 15 lambs distributed in the infected treatment (I, n=8) e control (C, n=7) as described above were used. After 66 days of the last inoculation was injected 6.6 MBq of 32P/lamb to evaluate the kinetics of the mineral. Blood, faeces and urine samples were collected in the following seven days and the slaughter of lambs was carried out on the last day in order to collect bone and soft tissues (Liver, kidney, heart and muscle) samples. To analyze P flows the biomathematical model described by Lopes et al. (2001) was used. Similar P intake between treatments was verified (VI) (P > 0.1), lower absorption of endogenous (Vaf) and dietary P (Vaa) as well as higher excretion of dietary P (VFD) in infected lambs (P < 0.1). With the lower absorption (VaT) of P in infected lambs, there was, consequently, lower distribution and lower incorporation of P in the bones (VO+D). It was concluded that P metabolism of lambs infected with T. colubriformis was altered, with reduced intestinal absorption and bioavailability, increased faecal loss and reduced P flow to bone

L'étude des interactions entre Oestrus ovis (Diptera : Oestridae) et Trichostrongylus colubriformis (Nematoda : Trichostronglidae) chez le mouton a montré qu' O. Ovis avait un effet négatif sur l'installation et la reproduction de T. Colubriformis. Le nombre de vers installés, leur fécondité et le nombre d'oeufs éliminés dans les matières fécales sont significativement diminués. En revanche, une infestation préalable par T. Colubriformis n'a pas d'influence sur une infestation ultérieure par O. Ovis. Ces parasites provoquent l'arrivée de nombreux mastocytes, globules leucocytes et éosinophiles dans les muqueuses parasitées. L'observation capitale de ce tarvail est la colonisation à distance par ces cellules des muqueuses respiratoires ou digestives qui ne sont pas le siège du parasite. O. Ovis induit toujours une éosinophilie sanguine et tissulaire qui sont précoces après l'infestation, mais dès l'élimination des larves elles diminuent rapidement.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
O estudo teve por objetivo determinar o grau de eficácia da levamisol, albendazol, ivermectina, moxidectina, closantel e triclorfon em isolados de Haemonchus contortus e Trichostrongylus colubriformis. Quarenta e dois cordeiros da raça Santa Inês, com três meses de idade, foram infectados artificialmente com 4000 larvas infectantes (L3) de H. contortus e 4000 L3 de T. colubriformis. Os animais foram separados em sete grupos, com seis animais cada, os quais receberam os seguintes tratamentos: Grupo 1 – controle, sem tratamento; Grupo 2 - moxidectina injetável (0,2 mg/kg de peso vivo (PV), Cydectin®, Fort Dodge), Grupo 3 - closantel via oral (10 mg/kg de PV, Zuletel®, Laboratório Microsules) Grupo 4 – triclorfon via oral (100 mg/kg de PV, Neguvon®, Bayer); Grupo 5 – fosfato de levamisol injetável (4,7 mg/kg de PV, Ripercol®, Fort Dodge), Grupo 6 - albendazol via oral (5,0 mg/kg de PV, Valbazen®, Pfizer) e o Grupo 7- tratado com ivermectina injetável (0,2 mg/kg de PV, Ivomec® , Merial). A via de administração e a dosagem empregada foram realizadas de acordo com as instruções do fabricante. Amostras de fezes foram coletadas no dia do tratamento, três, sete, 10 e 14 dias após, para a realização de contagem de ovos por grama de fezes (OPG) e cultura de fezes para obtenção e posterior identificação das L3. Os animais foram sacrificados 14 dias após o tratamento para obtenção e quantificação dos vermes presentes no abomaso e no intestino delgado. A eficácia dos tratamentos foi calculada a partir da média aritmética do OPG ou número total de vermes encontrados nos grupos tratados em comparação com os valores do grupo controle. As reduções percentuais das cargas parasitárias de H. contortus foram de 17% para o albendazol, 10% para o levamisol, 45% para moxidectina, 20% para ivermectina, 23% para o closantel e 73% para o triclorfon...
The objective of this study was to determine the efficacy of levamisole, albendazole, ivermectin, moxidectin, closantel and trichlorfon, against Haemonchus contortus and Trichostrongylus colubriformis isolates. Forty-two lambs of the Santa Ines breed, at three months of age, were simultaneously artificially infected with 4000 infective H. contortus larvae (L3) and 4000 T. colubriformis L3. The animals were divided into seven groups with six animals each that received one of the following treatments: Group 1 - control, no treatment; Group 2 - moxidectin (0.2 mg/kg body weight (BW), Cydectin®, Fort Dodge); Group 3 - closantel (10 mg/kg BW, 10% Zuletel®, Microsules Laboratories); Group 4 - trichlorfon (100 mg/kg BW, Neguvon®, Bayer); Group 5 – levamisole phosphate (4.7 mg/kg BW, Ripercol®, Fort Dodge); Group 6 - albendazole (5.0 mg/kg BW, Valbazen®, Pfizer); and Group 7 - treated with ivermectin (0.2 mg/kg BW, Ivomec®, Merial). Fecal samples were collected 3, 7, 10 and 14 days after treatment and processed for nematode fecal egg counts (FEC) and fecal cultures for the production and subsequent identification of L3. The animals were sacrificed 14 days after treatment for collection and quantification of the worms in the abomasum and small intestine. The efficacy of the treatments was calculated from the arithmetic mean of the FEC or worm burden of the treated groups, compared with the values of the control group. The percentage reductions in H. contortus worm burdens were 17% for albendazole, 10% for levamisole, 45% for moxidectin, 20% for ivermectina, 23% for closantel and 73% for trichlorfon. Reductions for T. colubriformis were 19% for albendazole, 28% for ivermectin, 82% for moxidectin and 0% for levamisole, closantel and trichlorfon. The fecal examination was not effective for the detection of resistance to T. colubriformis, since T. colubriformis L3 were not detected... (Complete abstract click electronic access below)

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Universidade Estadual Paulista (UNESP)
O experimento teve como objetivo avaliar a sobrevivência de larvas infectantes (L3) de Trichostrongylus colubriformis em três espécies forrageiras. Para tal, foram utilizados módulos experimentais constituídos por seis canteiros de 32,4 m2 cada, estabelecidos com as seguintes gramíneas forrageiras: Brachiaria decumbens cv. Australiana, Cynodon dactilon cv. Coast-cross e Panicum maximum cv. Aruana, perfazendo dois canteiros por espécie. Cada canteiro foi dividido em 36 parcelas, de 30 x 30 cm, de forma a permitir seis repetições por espécie e por altura em cada semana de colheita de material. A sobrevivência larval foi avaliada nas quatro estações do ano, sob o efeito de duas alturas de poda das forragens (baixa, 5 cm e alta, 30 cm). A poda foi realizada imediatamente antes da deposição das fezes. Ocorreram quatro deposições de fezes, uma a cada estação do ano. A colheita das fezes e da forragem foi realizada uma, duas, quatro, oito, 12 e 16 semanas após cada deposição de fezes nos canteiros experimentais. A altura da forragem foi medida em cada uma das subdivisões imediatamente antes das colheitas. A forragem foi cortada com uma tesoura de poda, rente ao solo, de uma área delimitada com o auxílio de um círculo de 10 cm de raio. As fezes foram recolhidas manualmente dos canteiros. A recuperação de L3 das amostras fecais foi maior quando as fezes foram depositadas em meio ao capim alto (com 30 cm - P<0,05). Porém, a recuperação de L3 nas forragens foi similar em ambos os cortes. Em relação à concentração de L3, o corte baixo propiciou maior quantidade de L3 por quilo de matéria seca (P<0,05). As maiores recuperações de L3, nas fezes e nas forragens, ocorreram quando as fezes foram depositadas no outono e na primavera. Dentre as espécies forrageiras, o capim aruana foi o que no geral apresentou maiores concentrações de larvas infectantes de T. colubriformis.
The purpose of the experiment was to evaluate infective Trichostrongylus colubriformis larvae (L3) survival in three forage species. Experimental modules formed by six 32.4-m2 plot, established with the following forage grass species, were used in the study: Australian Brachiaria decumbens cv., Cynodon dactilon cv. Coast-cross, and Panicum maximum cv. Aruana, totaling two plots for each species. Each plot was divided into 36 30 x 30 cm parts in order to allow six replicates per species and per height in each week of material collection. Larval survival was evaluated in the four seasons of the year under the effect of two forage paring heights (low, 5 cm, and high, 30 cm). The paring was carried out immediately before the feces were deposited. Four feces deposits were made, one in each season of the year. Feces and forage collection was performed one, two, four, eight, 12 and 16 weeks after each feces deposition in the experimental plots. Forage grass height was measured in each subdivision immediately before the collections. The forage was cut using pairing scissors, close to the soil, from an area delimited with a circle with a 10-cm radius. The feces were collected manually from the plots. L3 recovery rates from fecal samples were bigger when the feces were deposited in high grass (measuring 30 cm - P<0,05). However, L3 recovery in the forages was similar for both cuts. So far as the L3 concentration is concerned, the low cut resulted in a bigger number of L3 per kilogram of dry matter (P<0,05). The higher L3 recovery in the feces and in the forage grass occurred when the feces were deposited in the autumn and in the spring. Among the forage species, the aruana grass was the one that, in general, harbored the biggest concentrations of infecting T. colubriformis larvae.

Orientador: Alessandro Francisco Talamini do Amarante
Banca: Maria Conceição Zocoller Seno
Banca: Raquel A. da Rocha de Oliveira
Resumo: O estudo teve por objetivo determinar o grau de eficácia da levamisol, albendazol, ivermectina, moxidectina, closantel e triclorfon em isolados de Haemonchus contortus e Trichostrongylus colubriformis. Quarenta e dois cordeiros da raça Santa Inês, com três meses de idade, foram infectados artificialmente com 4000 larvas infectantes (L3) de H. contortus e 4000 L3 de T. colubriformis. Os animais foram separados em sete grupos, com seis animais cada, os quais receberam os seguintes tratamentos: Grupo 1 - controle, sem tratamento; Grupo 2 - moxidectina injetável (0,2 mg/kg de peso vivo (PV), Cydectin®, Fort Dodge), Grupo 3 - closantel via oral (10 mg/kg de PV, Zuletel®, Laboratório Microsules) Grupo 4 - triclorfon via oral (100 mg/kg de PV, Neguvon®, Bayer); Grupo 5 - fosfato de levamisol injetável (4,7 mg/kg de PV, Ripercol®, Fort Dodge), Grupo 6 - albendazol via oral (5,0 mg/kg de PV, Valbazen®, Pfizer) e o Grupo 7- tratado com ivermectina injetável (0,2 mg/kg de PV, Ivomec® , Merial). A via de administração e a dosagem empregada foram realizadas de acordo com as instruções do fabricante. Amostras de fezes foram coletadas no dia do tratamento, três, sete, 10 e 14 dias após, para a realização de contagem de ovos por grama de fezes (OPG) e cultura de fezes para obtenção e posterior identificação das L3. Os animais foram sacrificados 14 dias após o tratamento para obtenção e quantificação dos vermes presentes no abomaso e no intestino delgado. A eficácia dos tratamentos foi calculada a partir da média aritmética do OPG ou número total de vermes encontrados nos grupos tratados em comparação com os valores do grupo controle. As reduções percentuais das cargas parasitárias de H. contortus foram de 17% para o albendazol, 10% para o levamisol, 45% para moxidectina, 20% para ivermectina, 23% para o closantel e 73% para o triclorfon... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: The objective of this study was to determine the efficacy of levamisole, albendazole, ivermectin, moxidectin, closantel and trichlorfon, against Haemonchus contortus and Trichostrongylus colubriformis isolates. Forty-two lambs of the Santa Ines breed, at three months of age, were simultaneously artificially infected with 4000 infective H. contortus larvae (L3) and 4000 T. colubriformis L3. The animals were divided into seven groups with six animals each that received one of the following treatments: Group 1 - control, no treatment; Group 2 - moxidectin (0.2 mg/kg body weight (BW), Cydectin®, Fort Dodge); Group 3 - closantel (10 mg/kg BW, 10% Zuletel®, Microsules Laboratories); Group 4 - trichlorfon (100 mg/kg BW, Neguvon®, Bayer); Group 5 - levamisole phosphate (4.7 mg/kg BW, Ripercol®, Fort Dodge); Group 6 - albendazole (5.0 mg/kg BW, Valbazen®, Pfizer); and Group 7 - treated with ivermectin (0.2 mg/kg BW, Ivomec®, Merial). Fecal samples were collected 3, 7, 10 and 14 days after treatment and processed for nematode fecal egg counts (FEC) and fecal cultures for the production and subsequent identification of L3. The animals were sacrificed 14 days after treatment for collection and quantification of the worms in the abomasum and small intestine. The efficacy of the treatments was calculated from the arithmetic mean of the FEC or worm burden of the treated groups, compared with the values of the control group. The percentage reductions in H. contortus worm burdens were 17% for albendazole, 10% for levamisole, 45% for moxidectin, 20% for ivermectina, 23% for closantel and 73% for trichlorfon. Reductions for T. colubriformis were 19% for albendazole, 28% for ivermectin, 82% for moxidectin and 0% for levamisole, closantel and trichlorfon. The fecal examination was not effective for the detection of resistance to T. colubriformis, since T. colubriformis L3 were not detected... (Complete abstract click electronic access below)
Mestre

Orientador: Alessandro Francisco Talamine do Amarante
Banca: Solange Maria Gennari
Banca: Cledson Augusto Garcia
Banca: Reinaldo José da Silva
Banca: Patrizia Ana Bricarello
Resumo: O experimento teve como objetivo avaliar a sobrevivência de larvas infectantes (L3) de Trichostrongylus colubriformis em três espécies forrageiras. Para tal, foram utilizados módulos experimentais constituídos por seis canteiros de 32,4 m2 cada, estabelecidos com as seguintes gramíneas forrageiras: Brachiaria decumbens cv. Australiana, Cynodon dactilon cv. Coast-cross e Panicum maximum cv. Aruana, perfazendo dois canteiros por espécie. Cada canteiro foi dividido em 36 parcelas, de 30 x 30 cm, de forma a permitir seis repetições por espécie e por altura em cada semana de colheita de material. A sobrevivência larval foi avaliada nas quatro estações do ano, sob o efeito de duas alturas de poda das forragens (baixa, 5 cm e alta, 30 cm). A poda foi realizada imediatamente antes da deposição das fezes. Ocorreram quatro deposições de fezes, uma a cada estação do ano. A colheita das fezes e da forragem foi realizada uma, duas, quatro, oito, 12 e 16 semanas após cada deposição de fezes nos canteiros experimentais. A altura da forragem foi medida em cada uma das subdivisões imediatamente antes das colheitas. A forragem foi cortada com uma tesoura de poda, rente ao solo, de uma área delimitada com o auxílio de um círculo de 10 cm de raio. As fezes foram recolhidas manualmente dos canteiros. A recuperação de L3 das amostras fecais foi maior quando as fezes foram depositadas em meio ao capim alto (com 30 cm - P<0,05). Porém, a recuperação de L3 nas forragens foi similar em ambos os cortes. Em relação à concentração de L3, o corte baixo propiciou maior quantidade de L3 por quilo de matéria seca (P<0,05). As maiores recuperações de L3, nas fezes e nas forragens, ocorreram quando as fezes foram depositadas no outono e na primavera. Dentre as espécies forrageiras, o capim aruana foi o que no geral apresentou maiores concentrações de larvas infectantes de T. colubriformis.
Abstract: The purpose of the experiment was to evaluate infective Trichostrongylus colubriformis larvae (L3) survival in three forage species. Experimental modules formed by six 32.4-m2 plot, established with the following forage grass species, were used in the study: Australian Brachiaria decumbens cv., Cynodon dactilon cv. Coast-cross, and Panicum maximum cv. Aruana, totaling two plots for each species. Each plot was divided into 36 30 x 30 cm parts in order to allow six replicates per species and per height in each week of material collection. Larval survival was evaluated in the four seasons of the year under the effect of two forage paring heights (low, 5 cm, and high, 30 cm). The paring was carried out immediately before the feces were deposited. Four feces deposits were made, one in each season of the year. Feces and forage collection was performed one, two, four, eight, 12 and 16 weeks after each feces deposition in the experimental plots. Forage grass height was measured in each subdivision immediately before the collections. The forage was cut using pairing scissors, close to the soil, from an area delimited with a circle with a 10-cm radius. The feces were collected manually from the plots. L3 recovery rates from fecal samples were bigger when the feces were deposited in high grass (measuring 30 cm - P<0,05). However, L3 recovery in the forages was similar for both cuts. So far as the L3 concentration is concerned, the low cut resulted in a bigger number of L3 per kilogram of dry matter (P<0,05). The higher L3 recovery in the feces and in the forage grass occurred when the feces were deposited in the autumn and in the spring. Among the forage species, the aruana grass was the one that, in general, harbored the biggest concentrations of infecting T. colubriformis larvae.
Doutor

Orientador: Alessandro Francisco Talamini do Amarante
Resumo: Na produção animal o planejamento nutricional inadequado desencadeia grandes prejuízos ao sistema produtivo, trazendo consequências adversas aos animais e menores lucros ao criador. Este trabalho teve por objetivo avaliar a influência da suplementação alimentar sobre a imunidade dos cordeiros, biologia e patogenia de Haemonchus contortus e Trichostrongylus colubriformis. Quarenta e quatro cordeiros machos da raça Dorper com aproximadamente quatro meses e 31,5±3,24 kg de peso corporal foram alocados aleatoriamente em quatro dietas à base de feno triturado, Cynodon dactylon (cv. Tyfton), acrescido de 0%, 25%, 50% ou 75% de concentrado. Em cada dieta sete animais foram infectados com 1.000 larvas infectantes (L3) de H. contortus e 1.000 L3 de T. colubriformis a cada três dias ao longo de 12 semanas. Em cada dieta, quatro animais permaneceram como grupo controle, Não-Infectados. Os grupos experimentais foram: 0%-Infectado, 0%-Não-Infectado, 25%-Infectado, 25%-Não-Infectado, 50%-Infectado, 50%-Não-Infectado, 75%-Infectado e 75%-Não-Infectado. Quanto maior a suplementação, menor a contagem de ovos por grama de fezes (OPG) de H. contortus (P < 0,0001), porém sem influência sobre contagem de OPG de T. colubriformis. O número de fêmeas de H. contortus foi maior no grupo 0% infectado que nas demais dietas (P = 0,0052). Em nenhuma fase parasitária de T. colubriformis houve efeito da Dieta (P > 0,05). Animais da dieta 0%-Infectado tiveram menor volume globular (VG) que seu grupo controle... (Resumo completo, clicar acesso eletrônico abaixo)
Doutor

Orientador: Luis Octavio Regasini
Resumo: Um dos maiores problemas sanitários encontrados pela pecuária são os nematoides gastrintestinais (NGIs), os quais causam perdas significativas aos pecuaristas de pequenos ruminantes. O controle de NGIs é realizado com base na administração repetida e imprópria dos anti-helmínticos (AHs) convencionais, havendo uma crescente presença de resíduos no meio ambiente ou em produtos de consumo. Acima de tudo, o desenvolvimento de resistência em populações de NGIs tornou-se um problema mundial. A fim de preservar a saúde dos hospedeiros, bem como retardar a resistência, torna-se necessário identificar métodos alternativos ao uso de AHs, tais como o uso de plantas, seus extratos e substâncias. Pterogyne nitens Tul. (Fabaceae), árvore nativa brasileira, apresenta indicações de uso popular como vermífugo, bem como de diversas substâncias bioativas, tais como: ácidos fenólicos, flavonoides, alcaloides guanidínicos, terpenos e esteróis. O presente estudo teve como objetivo investigar os efeitos de extratos e substâncias de P. nitens em diferentes estágios do ciclo de vida de três isolados susceptíveis originários do Brasil e da França de duas espécies de NGIs, Haemonchus contortus e Trichostrongylus colubriformis. Foram preparados três extratos etanólicos a partir das folhas (EEL), frutos (EEFR) e flores (EEFL); e oito substâncias fenólicas isoladas [duas flavonas (sorbifolina e pedalitina), dois flavonóis (quercetina e rutina), um flavan-3-ol (ourateacatequina) e três ácidos fenólicos (ác... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: One of the considerable health problems encountered by livestock is gastrointestinal nematodes (GINs), as they cause significant losses to small ruminant breeders. The control of GINs is based on the repeated and improper administration of synthetic anthelmintics (AHs), causing an increase of environmental residues and in food. Furthermore, the development of resistance in GIN populations has become a worldwide problem. In order to preserve the health of the hosts, as well as reduce resistance, it became necessary to identify AH alternative methods, such as the use of medicinal plants and their extracts and compounds. Pterogyne nitens Tul. (Fabaceae), brazilian native tree, presents popular use as a vermifuge and several bioactive compounds, such as phenolic acids, flavonoids, guanidine alkaloids, terpenes, and sterols. The aim of the current study was to investigate the effects of extracts and compounds from P. nitens against the different stages of the life cycle of three susceptible isolates from Brazil and France of two GIN species, Haemonchus contortus, and Trichostrongylus colubriformis. Three ethanolic extracts were prepared from leaves (EEL), fruits (EEFR), and flowers (EEFL). Eight phenolic compounds were isolated [two flavones (sorbifolin and pedalitin), two flavonols (quercetin and rutin), one flavan-3-ol (ouratecatechin) and three phenolic acids (caffeic acid, ferulic acid, and gallic acid)]. In addition, two flavonoids (chrysin and morin) were obtained commercial... (Complete abstract click electronic access below)
Doutor

Les Nématodes gastro-intestinaux sont des parasites majeurs en élevage ovin. Actuellement, leur contrôle repose sur l'utilisation de molécules anthelminthiques. L'extension de résistances à ces molécules dans les populations de strongles rend urgente la recherche de solutions alternatives comme la sélection d'animaux résistants. Toutefois, la méconnaissance des mécanismes de la réponse immunitaire des ovins contre ces strongles reste un obstacle à son développement. La première partie de nos travaux a démontré une orientation Th2 claire lors d'infestation par Haemonchus contortus et plus équivoque lors d'infestation par Trichostrongylus colubriformis. Dans un second temps, les réponses immunes adaptatives de moutons de race sensible (INRA 401) et résistante (Barbados Black Belly) lors d'infestation par H. contortus ont été comparées. Des différences d'expression des gènes de l'IL-4, IL-5 et de l'IL-13, et d'éosinophilie sanguine ont été enregistrées entre les deux races.

This study was stimulated by a publication of Mr Mike Bosch (Bosch 2007) that Cereus jamacaru DC (Cactaceae) used by him on his farm is effective for gastrointestinal nematode control in livestock. This plant, widely known as Queen of the night, is a serious invasive weed. We evaluated this claim in in vitro assays and in vivo experiments in sheep experimentally infected with Haemonchus contortus and Trichostrongylus colubriformis. This is the first report of such an anthelmintic trial in livestock involving C. jamacaru. The first study was to repeat the farmer’s work under strictly controlled conditions. In vivo studies were conducted to determine the possible direct anthelmintic effects of C. Jamacaru on ovine gastrointestinal nematodes. Eighteen sheep were allocated to three groups and were infected with 4000 H. Contortus and 6000 T. Colubriformis infective larvae given in 3 divided doses over a period of three days. From day 0 until day 49 of the experiment, sheep were drenched once a week with fresh blended C. jamacaru plants with the core removed at the same (32.33 g/sheep) or double the dose (64.66 g/sheep) used by Mr Bosch. No negative effects of the double dose were observed during the period of the experiment. Faeces were collected twice a week for faecal egg count. Based on the in vivo experiments, C. Jamacaru was not effective in reducing H. Contortus and T. Colubriformis in sheep to the 70% reduction levels of the control treatments considered to be a useful reduction in FEC. Nevertheless, its in vivo activity was substantial at the higher dose and reduced the FECs by 65%. To investigate the matter further some additional experiments were carried out. Specimens of C. Jamacaru were collected and dried in the shade. For the phytochemical analysis and in vitro experiments, the dried material was milled to a fine powder, it was extracted with acetone and five fractions (hexane, butanol, water, chloroform and 35% water in methanol) were obtained by solvent-solvent fractionation. The chemical composition of the fractions and crude extract was analysed by thin layer chromatography using three solvent systems of varying polarity and pH. To detect the separated compounds, vanillin-sulphuric acid-methanol was sprayed on the chromatograms and heated at 110°C for optimal colour development. The antioxidant activity in plant extracts may influence the immune systems of animals and have an indirect effect. The antioxidant activity was therefore determined. For qualitative analysis of antioxidant activity, the 2,2, diphenyl-1-picrylhydrazyl (DPPH) assay on TLC plates was used as a test for the radical scavenging ability of the compounds present in the different extracts. In the DPPH qualitative analysis of antioxidant activity there were a number of antioxidant compounds present in some of the extracts and fractions but the activity appeared to be low. This observation was confirmed in the TEAC quantitative analysis of antioxidant activity. Even the fraction with the highest activity was about 8 times less active than trolox or ascorbic acid. It therefore appears that stimulation of the immune system by antioxidant activity does not explain the results found on the farm. The antibacterial and antifungal activities were determined against 4 bacteria (Staphylococcus aureus, Enterococcus faecalis, Escherichia coli and Pseudomonas aeruginosa) and 3 fungi (Aspergillus fumigatus, Candida albicans and Cryptococcus neoformans). The minimum inhibitory concentration (MIC) values for plant extracts varied from 0.04 to 2.5 mg/ml. E. coli was the most sensitive and the fungi were in general resistant to the extract and fractions. In general the activity was not very good, especially if the dosage in an aqueous system on the farm is considered. Antimicrobial activity also does not explain the results obtained. The nematocidal activity was evaluated by an egg hatch assay and larval development viability assay (LDVA) in vitro. The acetone extract inhibited egg hatchability and killed infective larvae of H. Contortus nematodes of sheep in a concentration-dependent manner. The percentage inhibition of egg hatching of the acetone extract and the butanol, chloroform, 35% water in methanol, hexane and water solvent-solvent fractions were 100%, 100%, 94%, 91%, 9% and 16%, respectively. The percentage inhibition of larval development of the acetone extract and chloroform, 35% water in methanol, hexane and water fractions were 93%, 84%, 49%, 85%, 31%, respectively. The chloroform fraction was significantly more active than all the other fractions (p<0.05). The extracts had in vitro activity against the nematodes. The activity was however much lower than the positive control, albendazole. It is disappointing that our results do not provide an explanation for the success obtained by Mr Bosch on his farm. Even if we do not understand how it works it may be useful to prepare suitable dosages (using low level technologies adaptable to rural conditions) for use by resource-poor rural communities where C. jamacaru occurs as an invasive weed. Copyright
Dissertation (MSc)--University of Pretoria, 2010.
Paraclinical Sciences
unrestricted

Parmi les nématodes gastro-intestinaux ayant un impact majeur sur la santé des ruminants Haemonchus contortus est l’espèce la plus pathogène. Actuellement, le contrôle des populations parasitaires repose sur les traitements anthelminthiques dont l’efficacité est limitée par l’émergence de parasites résistants. L'établissement de nouvelles stratégies de lutte (thérapeutiques ou vaccinales) repose actuellement sur l'identification de nouvelles cibles moléculaires. Ainsi, les gènes spécifiquement régulés lors de la phase précoce d’interaction du parasite H. contortus avec son hôte sont des cibles de choix. Afin d’identifier ces gènes, 4 banques d’H. contortus enrichies en ADNc spécifiquement exprimés au stade L4 (5 jours après infestation) ont été obtenues par la technique d’Hybridation Suppressive et Soustractive (SSH). Sur 400 clones criblés (dot-blot, RT-PCR), 51 clones d’intérêts ont été regroupés en 10 contigs. Les candidats possédant un peptide signal de sécrétion ont fait l’objet d’une étude plus approfondie. Les homologues de ces candidats ont été recherchés chez les deux autres principales espèces de strongles gastro-intestinaux (T. circumcincta et T. colubriformis) dans le but d’évaluer leur polymorphisme. La production et la purification des protéines recombinantes correspondant à ces candidats a également été réalisée afin de tester leur éventuel pouvoir immunogène
Gastro-intestinal nematodes such as Haemonchus contortus have a major impact on health of small ruminants world-wide. The control of infections remains largely based on anthelminthic treatments, but spreading of resistance has reduced their efficiency. An attractive solution would be the development of anti-nematode vaccines. Genes expressed during the early parasitic stage of H. contortus constituted our main targets. We have developed an approach based on SSH (Suppressive Subtractive Hybridization) technique and generated 4 subtracted cDNA libraries of H. contortus enriched in cDNA specifically expressed during L4 stage (five days post infection). 400 clones were analyzed by dot-blotand 51 clones regrouped in 10 contigs. All contigs were validated by RT-PCR. Homologues of candidates possessing a signal peptide were searched in T. colubriformis and T. circumcincta to evaluate their polymorphism. Recombinant proteins of theses candidates were produced and purified in order to know if they have a good vaccine potential with cross protection against two major gastro-intestinal nematodes

This thesis describes a series of three experiments designed to estimate the nutritional cost of the immune response to the gastrointestinal nematodes Trichostrongylus colubriformis and Teladorsagia circumcincta in sheep. For each experiment, animals were allocated hierarchically by liveweight into one of four groups that were either infected (group IF), similarly infected and concurrently immuno-suppressed with weekly intramuscular injections of 1.3mg kg liveweight (LW)-1 of methylprednisolone acetate (group ISIF), immuno-suppressed only (group IS) or remained as controls (group C). Body composition of all animals was estimated using x-ray computer tomography prior to infection and at the conclusion of each study with bodyweight and faecal nematode egg counts (FEC; eggs gram-1 of fresh faeces (epg)) measured along with blood samples taken for the determination of levels of serum proteins, phosphate and antibodies. In the first trial (Chapter 3), the nutritional cost of both the acquisition and maintenance of immunity to gastro-intestinal nematodes was investigated using immunologically naive 5-month-old lambs and immunologically competent 17-month-old ewes during infection with 2,000 and 4,000 L3 infective T. colubriformis larvae d-1, respectively (80 L3 T. colubriformis larvae kgLW-1 d-1). Profiles of FEC and comparative worm burdens at slaughter indicated an effective immune response was maintained in IF ewes and developed in IF lambs while successfully suppressed in both ISIF lambs and ISIF ewes and was confirmed by serum antibody titres. The typical reduction in voluntary feed intake as a consequence of infection was observed in IF lambs (0.30, p<0.001) but not in IF ewes, ISIF lambs or ISIF ewes, and appeared to be associated with L3 IgA. Gross efficiency of use of metabolizable energy (ME) for net energy (NE) deposition was reduced by 0.20 in lambs during acquisition of immunity and by 0.16 in ewes maintaining an established immunity. Infection in immuno-suppressed animals reduced efficiency by 0.05 and 0.15 for lambs and ewes. These findings allowed the hypothesis that the reduction in feed intake and nutrient utilization in young parasitized sheep is caused by physiological signalling associated with the acquisition phase of the host immune response to infection, rather than simply the damage caused by the parasite per se. The second trial (Chapter 4) investigated the influence of metabolizable protein (MP) supply on the metabolic disturbances associated with the acquisition phase of the immune response during infection with 2,000 L3 T. colubriformis d-1. Groups of lambs were offered either a low protein (L; 62g MP kgDM-1) or high protein diet (H; 95g MP kgDM-1). Patterns of total daily egg excretion indicated that an effective immune response was developed in HIF, but not LIF, HISIF nor LISF and was confirmed by comparative worm burdens. The proportionate reduction in feed intake in immunologically normal animals was reduced through the provision of additional protein, being 0.12 in HIF and 0.23 in LIF. Regardless of diet, infection did not cause a reduction in feed intake in immuno-suppressed animals (p>0.05). Infection proportionately reduced the gross efficiency of ME utilization in immunologically normal animals by 0.23 in HIF (p=0.09) and by 0.51 in LIF (p=0.01), but not in immuno-suppressed animals. Immuno-suppression did not suppress serum L3 IgA levels in seven of the eight HISIF and four of the eight LISIF animals. Furthermore, only four out of the eight immunologically normal animals from both the HIF and LIF groups displayed an L3 IgA response. Consequently, regardless of immuno-suppression treatment, animals were termed as IgA responders (HR or LR) or non-responders (HN or LN). Feed intake was proportionately reduced from day 22 by 0.15 in HR (p=0.03) and by 0.32 in LR (p=0.01), but was not significantly reduced in HN or LN. Gross efficiency of ME utilization was significantly reduced for LN animals only, being proportionately 0.59 (p<0.01). These findings allowed the conclusion that additional MP reduced the consequence of immunological signalling that was displayed in reduced feed intake and in nutrient utilization, both of which appeared to be associated with an IgA response. It is hypothesized that the lessening of nutritional disturbance observed in high protein and immuno-suppressed animals could be a consequence of altered physiological signalling during the immunological cascade. The third trial (Chapter 5) utilized lambs infected with the abomasal parasite T. circumcincta to explore the possibility that the reduction in feed intake and nutrient utilization is a universal phenomenon of the acquisition phase of the immune response to nematode parasites inhabiting different organs along the gastrointestinal tract. In addition, immunological changes at the site of parasite infestation in the abomasal mucosa were measured from serial biopsy tissue samples taken from a further twelve animals that were surgically fitted with an abomasal cannula and either infected (CIF) or concurrently infected and immuno-suppressed as described previously (CISIF). The development of immunity in IF animals was accompanied by a 0.17 proportional decrease in feed intake between days 15 to 28 of infection (p<0.05) and a 0.20 proportional reduction in nutrient utilization (p=0.07), none of which were observed in ISIF animals. While FEC and worm burdens indicated successful immuno-suppression in ISIF animals, both serum IgA and total antibody production were not reduced. The development of immunity in CIF was reflected in an increase in both mast cells and globule leukocytes in serial abomasal tissue biopsies, both of which were reduced in CISIF (p<0.01 for both). In serial biopsy tissue, immuno-suppression did prevent a rise in tissue IgA that was apparent in CIF animals (p<0.01) although these changes were not reflected in serum IgA levels. It appears that the alleviation of the reduction in feed intake and nutrient utilization in young lambs through the use of corticosteroid induced immuno-suppression may be a universal phenomenon for both intestinal and abomasal parasites, but the association with and/or role of IgA during infection with T. circumcincta is unclear. In summary, the reduction in feed intake and nutrient utilization in sheep during infection with both the abomasal nematode T. circumcincta and the small intestine nematode T. colubriformis appears to be associated with a component(s) of the acquisition phase of the host immune response, rather than, as conventionally assumed, the direct mechanical damage of the parasite per se. It is hypothesised that the nutritional disturbance as a consequence of infection in young lambs may be the result of pro-inflammatory cytokines involved in immunological signalling that may also be associated with the production of IgA, the effects of which can be reduced through the provision of adequate MP. These studies provide evidence that the immune response to gastrointestinal parasites is nutritionally costly to the animal and have implications for application of manipulations that are intended to promote the development of a strong immune reaction in high producing animals.

The development of goat breeding is largely impaired by losses due to gastrointestinal nematodes infection and the main control method, the use of synthetic anthelmintics face problems due to the growing selection of resistant populations of nematodes, making it imperative to search for new methods of control. The secondary metabolites present in several plants with bioactive properties represent a promising alternative for the control of worms parasites of small ruminants and may act directly or indirectly on the parasites. The aim of this study was to assess whether food supplementation with the powdered bark of Acacia mangium, a plant rich in condensed tannin (TC), would stimulate a pattern of immune response in goats, and consequently affect the experimental infection with Haemonchus contortus and Trichostrongylus colubriformis. This experiment was carried out with 18 goats, initially free of helminth infection, that were distributed in three groups: I - control (n = 6), II - Acacia (n = 6) and III - Acacia + PEG (n = 6). The animals in groups II and III received together with the concentrated (88% corn meal and 12% soybean meal) supplementation with bark of A. mangium (CT 100mg / kg BW), and the group III also received additional treatment of A. mangium with 10 g of polyethylene glycol. Fifteen days after the begin the administration of the plant, all animals were infected orally with a pool of 12,000 L3 larvae of H. contortus and T. colubriformis. After 35 days of infection the animals were euthanized and parasite burden of abomasum and thin intestine was determined. Blood samples and feces were collected for determination of immunological parasitological and parameters throughout the experiment. Our results showed that supplementation with A. mangium promoted changes in the immune profile of goats, inducing the increase of the number of circulating CD4 + T lymphocytes producing IL-4 and CD8 + T producing IL-4 and IFN-. Moreover, the supplementation with A. mangium demonstrated to be also effective for reducing the burden of parasitic adult worms of T. colubriformis despite it not prevented the establishment of infection and H. contortus and T. colubriformis. These results suggest that A. mangium may present bioactive property that induce an immune response associated with the control of helminth infections.
O desenvolvimento da caprinocultura sofre grandes prejuízos em decorrência das infecções por nematóides gastrointestinais e o principal método de controle, o uso dos anti-helmínticos sintéticos, enfrentam problemas devido à crescente seleção de populações de nematóides resistentes, tornando imprescindível a busca por novos métodos de controle. Os metabólitos secundários presentes em várias plantas com propriedades bioativas representam uma alternativa promissora para o controle da verminose dos pequenos ruminantes, podendo atuar de forma direta ou indireta sobre os parasitos. O objetivo deste trabalho foi determinar se a suplementação com o pó da casca da Acacia mangium, planta rica em Taninos condensados (TC), estimularia um padrão de resposta imune em caprinos, e consequentemente afetaria a infecção por Haemonchus contortus e Trichostrongylus colubriformis. Este experimento foi realizado com 18 caprinos, inicialmente livres de infecção helmíntica, que foram distribuídos em três grupos: I - controle (n=6), II - Acácia (n=6) e III - Acácia+PEG (n=6). Os animais dos grupos II e III receberam junto com o concentrado (88% farelo de milho e 12% farelo de soja) suplementação com pó da casca de A. mangium (100mg TC /Kg PV), e o grupo III recebeu também tratamento adicional de A. mangium com 10 g de polietilenoglicol. Após 15 dias do início da administração da planta todos os animais foram infectados por via oral com um pool de 12.000 larvas L3 de H. contortus e T. colubriformis. Após 35 dias de infecção os animais foram sacrificados e a carga parasitária de abomaso e intestino delgado foi avaliada. Ao longo de experimento amostras de sangue e fezes foram coletadas para determinação de parâmetros imunológicos e parasitológicos. Os resultados revelaram que a suplementação alimentar com A. mangium promoveu mudanças no perfil imunológico dos caprinos, aumentando o número de linfócitos TCD4+ produtores de IL-4 e TCD8+ produtores de IL-4 e IFN- circulantes; além disso, a suplementação com A. mangium também foi eficiente em promover a redução da carga parasitária de vermes adultos de T. colubriformis, apesar de não impedir o estabelecimento da infecção por H. contortus e T. colubriformis. Estes resultados demonstram que a A. mangium possui propriedades bioativas que induzem uma resposta imune associada ao controle de infecções helmínticas.

Tese (doutorado)—Universidade de Brasília, Faculdade de Agronomia e Medicina Veterinária, Programa de Pós-Graduação em Ciências Animais, 2011. Brasília, 2011.
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O tanino condensado (TC) no valor nutritivo de forragens temperadas e na saúde dos ruminantes em concentrações moderadas pode ser utilizado para promover aumento da eficiência da digestão da proteína e melhoria das condições sanitárias do rebanho, influenciando positivamente a qualidade da carne. Objetivou-se avaliar a interferência do TC presente no Sansão do Campo utilizado na dieta de cordeiros sobre as características quantitativas e qualitativas da carcaça e sua ação anti-helmíntica sobre T. colubriformis. Foram utilizados 24 cordeiros machos inteiros da raça Santa Inês com peso vivo (PV) 26,89±2,16kg criados em sistema de confinamento total desde o nascimento e mantidos em baias individuais por 10 semanas. Foi coletado dado do peso médio inicial e final, ganho total (GT), ganho médio diário (GMD), peso metabólico (PM), consumo em relação ao PV, consumo de matéria seca (MS), consumo de MS por unidade de PM e conversão alimentar. O delineamento experimental foi inteiramente casualizado com arranjo fatorial (2X2) com quatro tratamentos, com dois tipos de dietas com e sem Polietilenoglicol (PEG) e animais sadios e infectados. Análises foram feitas através do teste de médias (Tukey 5%) do programa estatístico SAS. Após o jejum e dieta hídrica, os animais foram abatidos e as carcaças permaneceram em câmara fria por um período de 24h. Foram coletados os dados referentes às características quantitativas e qualitativas da carcaça, dos componentes corporais e cortes comerciais. Realizada a leitura do pH inicial e final e avaliações subjetivas da distribuição de gordura na carcaça. Retiradas as frações da região da 11 a 13a costelas da carcaça esquerda para análise de área de olho de lombo, composição tecidual e centesimal, avaliação dos parâmetros físico-químicos e composição de ácidos graxos (AG). Houve diferença significativa (p<0,05) com valor superior da contagem de ovos por grama de fezes (OPG) para o tratamento Sansão+Infecção+PEG e um menor grau de infecção para o tratamento Sansão+Infecção. Para GMD e GT foi verificado que o tratamento Sansão+Infecção (116,0g e 7,5kg) apresentou valores inferiores aos tratamentos Sansão+PEG (150,7g e 9,8kg) e Sansão+PEG+Infecção (152,9g e 9,9kg) demonstrando redução do desempenho desses animais, mas não havendo diferença significativa entre o tratamento Sansão (142,5g e 9,2kg). Foi observada diferença significativa (p<0,05) para perímetro e o peso dos testículos, sendo que as maiores médias foram no tratamento Sansão+PEG (24,62cm e 0,40kg) quando comparada com as médias obtidas nos tratamentos Sansão+Infecção+PEG (22,00cm e 0,28kg), Sansão (21,00cm e 0,25kg) e Sansão+Infecção (22,23cm e 0,28kg). Características quantitativas, cortes comerciais, composição tecidual e centesimal, avaliação dos parâmetros físico-químicos apresentaram similaridade nos tratamentos. A dieta que contem TC presente no Sansão do Campo é recomendada por apresentar um desempenho semelhante nos animais sadios e infectados por desencadear uma redução do OPG de T. colubriformis em ovinos infectados, além de influenciar o perfil de AG saturados, mono e poli-insaturados. O ácido graxo oleico foi que mais contribuiu para a composição total dos ácidos graxos monoinsaturados e o ácido graxo araquidônico nos ácidos graxos poli-insaturados. _______________________________________________________________________________________ ABSTRACT
The condensed tannin (CT) in temperate forage nutritive value and health of ruminants in moderate concentrations can be used to promote increased efficiency of protein digestion and health conditions of the herd, positively influencing the quality of meat. The objective was to evaluate the interference of CT present in the Sansão do Campo in the diet of lambs on the quantitative and qualitative carcass and its anthelmintic action on T. colubriformis. We used 24 male lambs entire Santa Ines weight (BW) 26.89 ± 2.16 kg raised in total confinement system from birth and kept in individual stalls for 10 weeks. We collected data of initial and final weight, total gain (TG), average daily gain (ADG), metabolic weight (MW), PV in relation to consumption, consumption of dry matter (DM), DM intake per unit of PM and feed conversion. The experimental design was completely randomized design with factorial arrangement (2x2) with four treatments, with two types of diets with and without polyethylene glycol (PEG) healthy and infected animals. Analyses were performed using the test of means (Tukey 5%) of the SAS statistical program. After fasting and water diet, the animals were slaughtered and the carcasses remained in cold storage for a period of 24 hours. We collected data on quantitative and qualitative characteristics of the carcass, body components and the commercial cuts. Performed the reading of the initial and final pH and subjective evaluations of the distribution of fat in the carcass. Withdrawals from the region of fractions 11 to 13th ribs of the carcass left for the analysis of loin eye area, and proximate tissue composition, evaluation of physical and chemical parameters and composition of fatty acids (FA). There was a significant difference (p<0.05) with higher value of egg counts per gram of feces (EPG) for treating infection Sanson+Infection + PEG and a lower degree of infection to treat Infection+Sanson for ADG and TG were found that treatment Sanson+Infection (116.0 g and 7.5 kg) were lower than the Sanson+PEG treatment (150.7 g and 9.8 kg) and Sanson+PEG+Infection (152.9 g and 9.9kg) demonstrating reduced performance of these animals, but no significant difference between treatment Sanson (142.5 g and 9.2 kg). It thas no significant difference (p<0.05) for perimeter and the weight of the testes, and the averages were higher in the treatment Sanson+PEG (24.62 cm and 0.40 kg) compared with the averages of the Sanson+Infection+PEG treatment (22.00 cm and 0.28 kg), Sanson (21.00 cm and 0.25 kg) and Sanson+Infection (22.23 cm and 0.28 kg). Quantitative traits, commercial cuts, and proximate tissue composition, evaluation of physical and chemical parameters had similar treatments. The diet containing Sanson CT present in the field is recommended to present a similar performance in healthy and infected animals by triggering a reduction of the parasite EPG T. colubriformis in sheep infected, and influence the profile of fatty acids, saturated, monounsaturated and polyunsaturated. The fatty acid was oleic contributor to the overall composition of fatty acids and monounsaturated arachidonic in fatty acids polyunsaturated.

Les nématodes gastro-intestinaux (NGI) représentent un problème sanitaire majeur dans le monde entier en systèmes de production à l’herbe des élevages de moutons et de chèvres. La maîtrise de ces parasites est désormais compliquée par l'apparition de résistances aux anthelminthiques (AH) chez les nématodes. L'utilisation de plantes riches en tanins (PRT) comme AH non conventionnels est une des alternatives possibles au contrôle chimique de ces maladies parasitaires. Cependant, le mécanisme d'action de ces plantes sur les vers adultes demeure inconnu. Cet objectif a constitué le premier but de cette étude en particulier en essayant de déterminer quelle part jouent les effets directs et indirects dans l’action des PRT sur les populations de Nématodes adultes dans le tractus digestif des petits ruminants. Un effet direct correspond à une action de type pharmacologique sur la biologie, la structure et l’ultrastructure des nématodes liés à des composés biochimiques présents dans les PRT. L'hypothèse d’un effet indirect correspondrait une modification de la réponse cellulaire dans les muqueuses gastro-intestinales de l'hôte, affectant la biologie des NGI. Cette thèse est divisée en deux parties: 1) Deux expériences in vivo réalisées respectivement, au Mexique et en France, visant à déterminer les effets directs ou indirects sur la biologie des NGI liés à la consommation de PRT tel le tzalam (Lysiloma latisiliquum), une Légumineuse arbustive du Yucatan, le sainfoin (Onobrychis viciifolia) une Légumineuse fourragère tempérée, et ainsi que le quebracho(Schinopsis spp.), un extrait riche en tannins condensés Les agneaux ont été infestés par Haemonchus contortus (au Mexique) ou H. contortus et Trichostrongylus colubriformis (en France). La consommation de tzalam a provoqué une perturbation de croissance des vers ou de fertilité des femelles d’H. contortus. Selon la durée de consommation, le quebracho a réduit la population de H. contortus et T. colubriformis, et affecte également la fertilité de T. colubriformis. Le sainfoin a affecté la fécondité de H. contortus sans affecter celle de T. colubriformis. Le comptage de cellules effectrices dans les muqueuses pour évaluer l’implication d’un effet indirect n'a pas montré de différences significatives entre les lots recevant ou non des PRT. La seconde partie s’est fondée sur un test in vitro et deux essais in vivo chez des chèvres infestées par H. contortus pour mieux comprendre l'effet de la consommation de PRT (tzalam, au Mexique ou sainfoin, en France) sur la structure et l’ultrastructure d’H. contortus. Les principales altérations ont été trouvées dans la cuticule et la région céphalique des vers. Des agrégats de matériel végétal autour de la vulve n’ont été trouvé qu’in vitro. L’examen de l’ultrastructure des H. contortus a montré des signes de vacuolisation, surtout visible dans les tissus intestinaux et musculaires. Ces lésions suggèrent que des composés secondaires présents dans le feuillage des PRT peuvent être impliqués dans les fonctions vitales d’ H. contortus, telles que la mobilité, la nutrition et éventuellement la reproduction
Gastrointestinal nematodes (GIN) represent a major health problem worldwide in grazing sheep and goat production systems. The control of these parasites has been complicated by the emergence of nematodes which are resistant to the commercially available anthelmintics (AHs). The use of tannin rich plants (TRP), as non-conventional Ahs, represents an alternative for the control of these parasites. However, the mechanism of action of such plants against adult populations of GIN has not been determined. The objective of the study was to determine the direct and indirect effect of TRP against adult populations of GIN in small ruminants. A direct effect is considered to be any action against the biology, structure and/or ultrastructure of the nematodes which is similar to a chemical AH. The indirect effect is a modification of the immune response of the host at the level of the gastrointestinal mucosa which affects the biology of the GIN. This study is divided into two stages: First stage) Two in vivo experiments conducted in Mexico and France respectively, determined the direct and indirect effects on the biology of GIN due to the consumption of the TRP forage of tzalam (Lysiloma latisiliquum), sainfoin (Onobrychis viciifolia) as well as a tannin-rich quebracho extract (Schinopsis spp). Lambs were artificially infected with Haemonchus contortus (Mexico) or H. contortus and Trichostrongylus colubriformis (France). The consumption of tzalam affected the length and fertility of H. contortus females. Meanwhile, the quebracho extract reduced the population of H. contortus and T. colubriformis also affecting the fecundity of T. colubriformis. The sainfoin affected the fecundity of H. contortus without affecting T. colubriformis. The indirect effect was not evident. Second stage) An in vitro assay and two in vivo experiments with goats infected with H. contortus allowed to identify the effect of the consumption of TRP (tzalam or sainfoin) on the structure and ultrastructure of H. contortus. Alterations in cuticle and cephalic region were found. The aggregates in the vulva were only found in vitro. The ultrastructure of H. contortus showed a vacuolization process in the intestinal and muscular tissues of these nematodes. The lesions suggest that the compounds contained in the foliage of TRP may be involved in vital functions of H. contortus such as mobility, nutrition and possibly reproduction. This study is a contribution towards the understanding the mechanisms of AH action of the TRP against GIN

A reduction in voluntary feed intake is a major factor in the lost productivity of grazing lambs infected by gastrointestinal parasites yet the mechanisms involved are poorly understood. Potential pathways involved in parasite-induced feed intake depression were investigated in lambs with minimal previous exposure to parasites and artificially infected by the small intestinal parasite Trichostrongylus colubriformis. Six in vivo experiments were conducted on lambs housed in individual pens or metabolism crates with similar feeding and experimental procedures. In Experiment 1 (Chapter 4) the effect of T. colubriformis infection on short term feed intake in lambs and of some pharmacological agents on feed intake depression were investigated. Prior to and for the duration of infection, lambs were fed once per day and feed intake recorded at regular intervals over the day (8 h). Following the onset of feed intake depression in the infected group (9 weeks after commencing dosing), all animals were treated with an analgesic (codeine phosphate per os), an anti-inflammatory agent (indomethacin per os), a CCK antagonist (L364-718 by subcutaneous injection) or saline (control) in a replicated Latin square design (n = 8). Although the pattern of feed consumption was similar in infected and non-infected lambs, average daily intake was reduced 32 % and short term intake (recorded at 10 minute intervals for the first hour of feeding, 15 minute intervals for the second hour and hourly for the next 6 hours of feeding) reduced 40 % by infection. This identified the key component by which intake was depressed and enabled the use of a short term intake model and short duration of action compounds to identify the pathways involved in intake depression in this sequence of experiments. None of the pharmacological treatments increased intake in the infected group. These results suggest a reduction in the rate of consumption due to reduced hunger signals, rather than change of meal eating patterns, is the major cause of feed intake depression. Specific conclusions about the pathways investigated using the pharmacological agents could not be obtained. Experiment 2 (Chapter 5) was designed to investigate the roles of pain and osmolality on feed intake depression. Digesta samples collected prior to and during parasite infection and before and after feeding had similar osmolalities (240-260 mosmol/l) which indicated that feeding or infection had no effect on osmolality of digesta. Following the onset of feed intake depression in infected animals, all animals were treated in a Latin square design (n = 4) with no treatment, saline, local anaesthetic (xylocaine) or analgesic (codeine phosphate) solution 15 minutes before feeding, by slow injection into the duodenum. There was no effect of these treatments on food intake. In the second part of the experiment, hyperosmotic solutions (mannitol and NaCI) markedly depressed short term intake in non-infected animals, suggesting a role for osmoreceptors in intake regulation. However these effects were not blocked by local anaesthetic so the depressed intake may have resulted from generalised malaise rather than from specific osmoreceptor effects. In Experiment 3 (Chapter 6) the role of peripheral CCK on intake depression was examined by a dose-response study utilising the CCK antagonist, loxiglumide. Intravenous injection of 5, 10 or 20 mg/kg LW of loxiglumide to infected lambs 10-15 minutes before feeding (n = 6) had no effect on feed intake at any of the dose levels. In experiment 4 (Chapter 7) loxiglumide was infused intravenously for 10 minutes (30 mg/kg/h) before feeding and for the first 2 h (10 mg/kg/h) after feed was offered to minimise any effect of the rate of clearance of loxiglumide on the lack of feed intake response. As well, the rate of marker disappearance from the abomasum was recorded in both infected and non-infected animals. Continuous infusion of loxiglumide did not attenuate parasite induced intake depression nor did it have any effect on abomasal emptying. Abomasal volume was reduced by infection (66.3 vs 162 ml) as was the fractional outflow rate (2.2 vs 2.8 ml/min) but these differences were accounted for by the lower level of feed intake in the infected animals. In Experiment 5 (Chapter 8) brotizolam, a benzodiazepine appetite stimulant, thought to act on the hypothalamus, was administered in a dose-response study to infected and non-infected animals (n = 4) immediately prior to feeding or following termination of the first meal (45 minutes after feeding) and the feed intake response recorded. Brotizolam elevated both the short term (0-0.75 h), daily (22 h) intake and all time intervals in the first 5 h after feeding in infected and non-infected animals when administered after the first meal but when administered prior to feeding elevated intake only over the first 6 h of feeding. In both cases the magnitude of the response was greater in infected animals than in non-infected animals. Brotizolam appeared to increase the rate of eating without having a major impact on meal eating patterns when administered before feeding. Where administration was after the first meal, the effect was due to an "extra" meal being consumed. These findings showed that infected animals can respond to central stimulators of intake although the mechanism of the response is not known. Opioids were implicated in intake depression as the rate of intake rather than meal patterns appeared to be the major parameter depressed under parasitism. This was examined in experiment 6 (Chapter 9) where animals (n = 6) were fasted for 26 h or not fasted, then treated with saline (control), brotizolam (intake stimulant) or naloxone (opioid antagonist) immediately prior to feeding. Fasting stimulated feed intake in the short term (100 % increase in 75 min) and over the day (12 % increase) in both infected and non-infected animals. Following fasting, infected animals ate a similar amount of feed to the non-infected, fasted animals and more than the non-infected, non-fasted animals. The signals resulting from a one day fast were sufficient in the short term to override parasite induced mechanisms causing feed intake depression. Naloxone suppressed the intake stimulatory effects of a 26 h fast in both infected and non-infected animals, which supports a role for endogenous opioids as hunger signals. Where animals were not fasted, naloxone reduced intake only in the non-infected animals which suggested endogenous opioid levels may be lower in infected animals than in non-infected animals. In the final experiment (Experiment 7, Chapter 10) the role of central hunger and satiety mechanism were investigated. Infected and non-infected animals (n = 6) were treated with naloxone or saline by intravenous injection, or saline and met-enkephalinamide (an opioid analogue) by intracerebral infusion, or naloxone and the opioid analogue simultaneously to investigate the role of central opioids in feed intake depression. To determine the role of CCK induced satiety signals on feed intake at a central level, loxiglumide and CCK were infused separately and in combination for 30 minutes prior to feeding and for the first 60 minutes of feed on offer, into a lateral cerebral ventricle of the brain of infected and control animals (n = 6). The opioid analogue tended to increase intake in infected animals but the effect was not significant probably because the dose used was too low to elicit a response in sheep. Naloxone depressed intake only in the infected animals, which conflicted with the results of Experiment 4. As a consequence these results were inconclusive because of the single low dose of opioid analogue used and the conflicting naloxone responses. CCK alone depressed intake by 39-52 % only in infected animals and this effect of the 90 minute infusion was evident over the 8 h short term recording period. Loxiglumide attenuated the feed intake depressive effects of CCK in the infected animals to the extent that intake was elevated above control levels. Loxiglumide alone was an intake stimulant in both infected and non-infected animals. Intake was increased over the entire 8 h but mostly in the second hour when intake was increased by 188 % in infected animals and by 16 % in the non-infected animals and resulted in almost continuous eating. These results showed loxiglumide will temporarily block the effect of parasite infection on feed intake in sheep when administered centrally and the fact that it blocked the effects of exogenous CCK on intake indicated that the effect is mediated via CCK receptors. In conclusion GIT parasite infection reduced both short term and daily feed intake apparently by a change in rate of intake rather than any alteration in meal patterns. It was further suggested that anyone of a number of potential peripheral pathways, including changes to osmolality, gut emptying, pain and inflammation of the gut, alone is not involved in anorexia in sofar as the compounds used could block these factors and the results support the idea that intake depression is mediated via a central mechanism. Intake in infected animals responded to a much greater extent when fasting, i.c.v. loxiglumide or brotizolam were employed. Feed intake thus appears to be regulated through the same mechanisms in infected and non-infected animals. The results from compounds affecting the central mechanism suggest central CCK receptors are important in parasite induced anorexia, possibly by changing the onset of satiety or by interacting with endogenous opioids to reduce the rate of feed intake. Secondly reduced endogenous opioids may be causing the reduction in the rate of feed consumption alone or as a result of other interactions. It was concluded that intake in parasitised animals could be increased to that of control animals by employing procedures and compounds thought to act on the hypothalamus.

Les nématodes gastrointestinaux (NGIs) représentent une contrainte majeure en élevage des ruminants. Jusqu’à présent, la maîtrise de ces parasitoses a reposé essentiellement sur l’emploi répété de traitements anthelminthiques (AH) de synthèse. Cependant, le développement constant de résistances aux AH dans les populations de nématodes chez les petits ruminants conduit à rechercher des méthodes alternatives ou complémentaires de lutte contre ces parasitoses. Au cours des deux dernières décennies, les données se sont accumulées montrant que certaines plantes étaient dotées de propriétés anthelminthiques, ce qui a été associé à la présence de tannins condensés (TCs) et de flavonoïdes. Ces ressources contenant des TCs, exploitées comme nutricaments, représentent donc une alternative attractive à la chimiothérapie, en combinant des effets bénéfique sur la santé et la nutrition des ruminants. Toutefois, une des difficultés à résoudre, pour un usage pertinent de ces nutricaments, est de comprendre l’origine de la variabilité des résultats observés dans l’activité AH. A côté de facteurs quantitatifs, la qualité (la structure) des TC semble également jouer un rôle dans les propriétés antiparasitaires mais ce facteur demeure peu exploré. En utilisant Haemonchus contortus comme modèle expérimental de nématode et le sainfoin (Onobrychis viciifolia) comme modèle de plantes contenant des tannins, l’objectif général de cette thèse a été de mieux comprendre les relations structure/activité des TCs. Les objectifs spécifiques des diverses études menées ont été 1) d’ examiner in vitro (par la méthode LEIA) l’activité AH de 36 fractions purifiées de TCs présentant une large diversité de caractéristiques structurelles [ex: la taille (mDP), les proportions prodelphinidols /procyanidols (PD/PC) et trans/cis ], et 2) le rôle possible d’interactions entre tannins et flavonoïdes, 3) d’ évaluer et comparer in vivo chez des agneaux, les conséquences sur les populations de vers et sur la résilience de l’hôte de la distribution de 2 ressources contenant des TCs de qualité contrastée pour les valeurs PD/PC [fort PD/PC, sainfoin (O. viciifolia); bas PD/PC, pellicules de noisettes (Corylus avellana)], 4) d’ examiner si les résultats varient en fonction de l’espèce et de la localisation des parasites (abomasum, H. contortus vs intestin grêle, Trichostrongylus colubriformis), 5) d’évaluer les concentrations, la qualité et la bio-disponibilité des TCs le long du tube digestif chez les ovins. Les résultats acquis ont montré: 1) des relations in vitro entre certaines caractéristiques des TCs et l’activité AH : un rapport élevé PD/PC pour les 2 espèces de nématodes auquel s’ajoute un rôle de la taille des tanins (mDP) pour H. contortus sont les facteurs identifiés. De manière générale, H. contortus s’est avéré plus sensible (valeurs EC50 plus faibles) aux effets des diverses ressources que T. colubriformis. Des synergies d’effets AH ont été observés entre tannins et flavonoides (quercétine et lutéoline), tout particulièrement dans le cas de TCs avec un faible rapport PD/PC. Dans l’étude in vivo, les agneaux consommant du sainfoin ont montré des effets AHs et une amélioration de la résilience de l’hôte alors que les effets de la noisette ont été beaucoup plus discrets. Les analyses menées sur les contenus des divers organes digestifs et les fèces, pour les deux types de ressources ont montré de fortes réductions de TCs détectés par rapport aux teneurs initiales dans la ration alors que les caractéristiques de structure étaient globalement préservées au long du tube digestif. En conclusion, le rôle de la qualité (structure) des TCs dans l’activité AH a été confirmé à la fois par les études in vitro et in vivo. Ces résultats suggèrent l’importance de prendre en compte ces facteurs dans la recherche de ressources pouvant être exploitées comme futurs nutricaments chez les ruminants.

Chez les petits ruminants, la maîtrise actuelle du parasitisme par des nématodes gastro-intestinaux repose sur l'emploi d'anthelminthiques de synthèse. En raison du développement de résistances à ces molécules, l'utilisation de plantes riches en tannins condensés est désormais explorée comme solution alternative aux anthelminthiques, mais le mode d'action en reste mal connu. L'objectif de cette thèse était de définir les mécanismes d'action des tannins condensés sur les larves infestantes des trichostrongles digestifs. Nous avons d'abord confirmé que la consommation d'une plante riche en tannins condensés affecte l'installation des larves chez l'hôte, puis montré que les perturbations concernent les deux étapes-clefs de l'installation des larves : le dégainement puis la pénétration dans les muqueuses. La mise en évidence de lésions ultra-structurales et des sites de fixation des tannins condensés chez les larves est venu corroborée les résultats des études fonctionnelles, suggérant un mécanisme d'action direct. L'analyse des principaux facteurs modulant les effets constatés a montré qu'ils sont non-spécifiques de l'espèce de nématode mais varie selon la forme des larves (engainées ou dégainées). Par ailleurs, la concentration et la nature des tannins condensés sont les deux principaux facteurs liés à la plante influençant les propriétés anthelminthiques contre les larves. Des hypothèses expliquant les interactions entre les tannins et les molécules des larves infestantes sont proposées
In small ruminants, the current control of gastrointestinal parasitism relies on the use of chemical anthelmintics. However, due to the increasing development of resistances to chemical anthelmintics, the potential use of condensed tannin-rich plants represents an alternative solution to the chemical treatments, although the mode of action remains unknown. The objective of the current work was therefore to define the mechanisms of actions of condensed tannins on the infective larvae of gastrointestinal nematodes. First, we have confirmed that the consumption of a condensed tannin-rich plant decreased the establishment of infective larvae in the host. In a second step, it has been shown that both the exsheathment and the penetration of the larvae in the host mucosae were disturbed after contact with condensed tannins. The observations of the ultra-structural modifications and the fixation of condensed tannins on the larvae have confirmed a direct mode of action. In addition, our results have shown that the effects of condensed tannins were non specific of the nematode species but that differences in susceptibility exist between the ensheathed and exsheathed larvae. The effects were also dependent of the concentration and the biochemical nature of the condensed tannins. Finally, hypotheses on the potential interactions at the molecular level between the condensed tannins and the infective larvae are suggested

This thesis describes a series of three experiments designed to estimate the nutritional cost of the immune response to the gastrointestinal nematodes Trichostrongylus colubriformis and Teladorsagia circumcincta in sheep. For each experiment, animals were allocated hierarchically by liveweight into one of four groups that were either infected (group IF), similarly infected and concurrently immuno-suppressed with weekly intramuscular injections of 1.3mg kg liveweight (LW)⁻¹ of methylprednisolone acetate (group ISIF), immunosuppressed only (group IS) or remained as controls (group C). Body composition of all animals was estimated using x-ray computer tomography prior to infection and at the conclusion of each study with bodyweight and faecal nematode egg counts (FEC; eggs gram⁻¹ of fresh faeces (epg)) measured along with blood samples taken for the determination of levels of serum proteins, phosphate and antibodies. In the first trial (Chapter 3), the nutritional cost of both the acquisition and maintenance of immunity to gastro-intestinal nematodes was investigated using immunologically naive 5-month-old lambs and immunologically competent 17-month-old ewes during infection with 2,000 and 4,000 L3 infective T. colubriformis larvae d⁻¹, respectively (80 L3 T. colubriformis larvae kgLW⁻¹ d⁻¹). Profiles of FEC and comparative worm burdens at slaughter indicated an effective immune response was maintained in IF ewes and developed in IF lambs while successfully suppressed in both ISIF lambs and ISIF ewes and was confirmed by serum antibody titres. The typical reduction in voluntary feed intake as a consequence of infection was observed in IF lambs (0.30, p<0.001) but not in IF ewes, ISIF lambs or ISIF ewes, and appeared to be associated with L3 IgA. Gross efficiency of use of metabolizable energy (ME) for net energy (NE) deposition was reduced by 0.20 in lambs during acquisition of immunity and by 0.16 in ewes maintaining an established immunity. Infection in immuno-suppressed animals reduced efficiency by 0.05 and 0.15 for lambs and ewes. These findings allowed the hypothesis that the reduction in feed intake and nutrient utilization in young parasitized sheep is caused by physiological signalling associated with the acquisition phase of the host immune response to infection, rather than simply the damage caused by the parasite per se. The second trial (Chapter 4) investigated the influence of metabolizable protein (MP) supply on the metabolic disturbances associated with the acquisition phase of the immune response during infection with 2,000 L3 T. colubriformis d⁻¹. Groups of lambs were offered either a low protein (L; 62g MP kgDM⁻¹) or high protein diet (H; 95g MP kgDM⁻¹). Patterns of total daily egg excretion indicated that an effective immune response was developed in HIF, but not LIF, HISIF nor LISF and was confirmed by comparative worm burdens. The proportionate reduction in feed intake in immunologically normal animals was reduced through the provision of additional protein, being 0.12 in HIF and 0.23 in LIF. Regardless of diet, infection did not cause a reduction in feed intake in immuno-suppressed animals (p>0.05). Infection proportionately reduced the gross efficiency of ME utilization in immunologically normal animals by 0.23 in HIF (p=0.09) and by 0.51 in LIF (p=0.01), but not in immuno-suppressed animals. Immuno-suppression did not suppress serum L3 IgA levels in seven of the eight HISIF and four of the eight LISIF animals. Furthermore, only four out of the eight immunologically normal animals from both the HIF and LIF groups displayed an L3 IgA response. Consequently, regardless of immunosuppression treatment, animals were termed as IgA responders (HR or LR) or non-responders (HN or LN). Feed intake was proportionately reduced from day 22 by 0.15 in HR (p=0.03) and by 0.32 in LR (p=0.01), but was not significantly reduced in HN or LN. Gross efficiency of ME utilization was significantly reduced for LN animals only, being proportionately 0.59 (p<0.01). These findings allowed the conclusion that additional MP reduced the consequence of immunological signalling that was displayed in reduced feed intake and in nutrient utilization, both of which appeared to be associated with an IgA response. It is hypothesized that the lessening of nutritional disturbance observed in high protein and immuno-suppressed animals could be a consequence of altered physiological signalling during the immunological cascade. The third trial (Chapter 5) utilized lambs infected with the abomasal parasite T. circumcincta to explore the possibility that the reduction in feed intake and nutrient utilization is a universal phenomenon of the acquisition phase of the immune response to nematode parasites inhabiting different organs along the gastrointestinal tract. In addition, immunological changes at the site of parasite infestation in the abomasal mucosa were measured from serial biopsy tissue samples taken from a further twelve animals that were surgically fitted with an abomasal cannula and either infected (CIF) or concurrently infected and immuno-suppressed as described previously (CISIF). The development of immunity in IF animals was accompanied by a 0.17 proportional decrease in feed intake between days 15 to 28 of infection (p<0.05) and a 0.20 proportional reduction in nutrient utilization (p=0.07), none of which were observed in ISIF animals. While FEC and worm burdens indicated successful immunosuppression in ISIF animals, both serum IgA and total antibody production were not reduced. The development of immunity in CIF was reflected in an increase in both mast cells and globule leukocytes in serial abomasal tissue biopsies, both of which were reduced in CISIF (p<0.01 for both). In serial biopsy tissue, immuno-suppression did prevent a rise in tissue IgA that was apparent in CIF animals (p<0.01) although these changes were not reflected in serum IgA levels. It appears that the alleviation of the reduction in feed intake and nutrient utilization in young lambs through the use of corticosteroid induced immuno-suppression may be a universal phenomenon for both intestinal and abomasal parasites, but the association with and/or role of IgA during infection with T. circumcincta is unclear. In summary, the reduction in feed intake and nutrient utilization in sheep during infection with both the abomasal nematode T. circumcincta and the small intestine nematode T. colubriformis appears to be associated with a component(s) of the acquisition phase of the host immune response, rather than, as conventionally assumed, the direct mechanical damage of the parasite per se. It is hypothesised that the nutritional disturbance as a consequence of infection in young lambs may be the result of pro-inflammatory cytokines involved in immunological signalling that may also be associated with the production of IgA, the effects of which can be reduced through the provision of adequate MP. These studies provide evidence that the immune response to gastrointestinal parasites is nutritionally costly to the animal and have implications for application of manipulations that are intended to promote the development of a strong immune reaction in high producing animals.

The series of experiments described in this thesis were designed to investigate the role of suckling or late weaning in the response of young lambs to nematode infection. All experiments were conducted outdoors with grazing animals and no supplementation but for suckled groups of lambs whose counterparts were weaned to ryegrass – white clover swards. The parasite of interest was mainly Teladorsagia circumcincta solely but with mixed infection of Trichostrongylus colubriformis in one instance. In Chapter 3 (first experiment), the hypothesis that milk per se may have a direct effect on nematode development, rather than an indirect effect through enhancement of host immunity by superior nutrient supply was tested. Sixty, twinborn lambs were used, allocated to one of eight groups formed by either dosing lambs from 42 days of age or not with the equivalent of 1000 or 250 L₃ T. circumcincta larvae d⁻¹ until five days before necropsy, while a twin was either weaned at 39 days of age, suckled as single or twin until necropsy on day 84. The possibility that weaning one of a twin set onto pasture in close proximity to the ewe would cause abnormal ewe and lamb behaviour was tested by replicating the work in twins maintained as twins but in which one twin received equivalent of 250 and the other 1000 L₃ T. circumcincta larvae d⁻¹. This showed no abnormal ewe nursing or lamb suckling behaviour as a result of weaning a twin in a set. Relatively low faecal egg counts (FEC) and a two to three fold lower worm burdens suggest suckling could reduce larval establishment. Inability to detect peripheral titres of immunoglobulins supports this conclusion. An intra worm-population regulation of T. circumcincta, indicated by a pattern of greater egg-laying by a numerically smaller but physiologically better developed nematode population in suckled lambs measured in eggs 'in utero' and worm length made interpretation of FEC difficult. Suckling significantly improved weight gain and carcass weights, but early weaning did not reduce resilience to infection. In Chapter 4 (second experiment), 40 pairs of twin lambs, average age of 39 days, were either infected with the equivalent of 1000 L₃ T. circumcincta larvae d⁻¹ or not, while one twin was weaned and the other allowed to continue suckling. Necropsy was carried out on groups of five and six lambs from each of the uninfected and infected treatments, respectively, at mean age of 84, 112, and on six lambs from each group at 140 days of age. This serial slaughter allowed further confirmation of the hypothesis in Chapter 3 but also investigated the long-term effect of suckling on resistance or resilience of lambs at the trial when immune responses were anticipated to be developing. An in vitro direct larval challenge (IVDC) study, to monitor larval establishment, was carried out on tissue explants from necropsied lambs. Suckled lambs consistently showed lower FEC (P < 0.05) and worm burdens (P < 0.05) at every phase of the trial. Within the infected groups, % in vitro larval rejection suggested earlier immune responses in the weaned lambs by day 84, which was not consistent with lower worm burdens in suckled lambs but appeared similar in the subsequent necropsies. Lambs continued to show better growth due to suckling while weaning did not reduce the resilience of lambs confirming observations in Chapter 3. The immunoglobulin profile suggested the commencement of immune responses in lambs from the period after the 84th day necropsy, with significantly greater (P < 0.01) IgA titre in the infected groups, and the suckled lambs towards the end of the trial on day 140. A vaccinating effect of early exposure to parasites was coincidentally revealed as a result of unintentional pasture larval contamination, seen in suckled non-infected lambs shedding fewer eggs and harbouring fewer worms during the later necropsies compared with their weaned non-infected counterparts. In Chapter 5 (third trial), 93 pairs of twin lambs, 47 pairs of which received a vaccinating mixed infection of T. circumcincta and T. colubriformis larvae (60 L₃ / kg W / d) at ratio 40:60, respectively during the period 36 – 103 days of age, were either weaned early on day 51 or later on day 108. All lambs were drenched on day 108 and groups received challenge infections from day 116, at same rate with the vaccinating infection, or not, which ceased five days before respective necropsies. Necropsies were carried out on selected lambs on days 108, 184 and 218. The direct effect of milk on larval establishment appeared to feature only in the T. circumcincta populations on slaughter day 108. The long-term benefit of late weaning for development of resistance was conditional on lambs receiving the vaccinating infection, and appeared to be more pronounced in the small intestine, reflected by a greater reduction of T. colubriformis populations in that organ than of T. circumcincta populations in the abomasum. A negative consequence of enhanced immune response was the suggestion of an increased metabolic cost in reduced performance of lambs. In conclusion, the work provides support to the hypotheses that: (a.) suckling may reduce the establishment of nematode larvae through the direct effect of milk, (b.) may enhance rapid development of host immunity to infection, and (c.) it further suggests that lack of larval experience during suckling may have long term negative implications for host resistance. Finally, it suggests that milk may play little role in the enhancement of host resilience to infection and, on the contrary, that additional metabolic cost may be associated with a more rapid development of immunity resulting from larval challenge while suckling.

Two studies were conducted to investigate anthelmintic resistance in goat parasites in New Zealand. In Study 1 parasites from goats on a farm with a long history of problems with anthelmintic efficacy were used to infect sheep for a controlled slaughter study. Nineteen lambs were acquired, effectively drenched and housed. Each was infected with a mixture of larvae comprising Haemonchus contortus, Teladorsagia circumcincta, Trichostrongylus colubriformis and Oesophagostomum venulosum. After 28 days lambs were restrictively randomised into 3 groups based on faecal egg counts. Group 1 was left untreated (n=6), Group 2 (n=6) was given a single dose of abamectin (0.2mg/kg) + levamisole HCL (8mg/kg) + oxfendazole (4.5mg/kg) (“Matrix Oral Drench for Sheep”®, Ancare, New Zealand) and Group 3 (n=7) was treated at twice the dose rate of Group 2. Fourteen days after treatment all animals were killed for total worm counts. The mean burdens of T. circumcincta in Group 1 was 337, in Group 2 was 68 (efficacy 80%) and in Group 3 was 10 (efficacy 97%). The mean burdens of T. colubriformis in Group 1 was 375, in Group 2 was 220 (efficacy 41%) and in Group 3 was 81 (efficacy 78%). Although the worm burdens in these lambs were low, all animals were infected with each of these two species except for T. circumcincta in Group 3 where only 3 lambs were infected. Efficacy against other species was 100%. These results clearly indicate that a single dose of a combination drench was ineffective against two species and even when a double dose was used the efficacy against T. colubriformis was only 78%. In Study 2 a survey of drench efficacy was conducted on 17 goat farms using the DrenchRite® larval development assay. Evidence of concurrent resistance to benzimidazoles, levamisole and ivermectin was detected in T. colubriformis and T. circumcincta on 11/17 and 3/14 respectively. Only 5 of 14 farms had previously undertaken some form of testing for drench resistance prior to this survey. Evidence from these two studies suggests that severe anthelmintic resistance is common on goat farms in New Zealand