Made available in DSpace on 2014-06-11T19:33:25Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-06-21Bitstream added on 2014-06-13T20:05:15Z : No. of bitstreams: 1 domingues_lf_dr_jabo.pdf: 549039 bytes, checksum: d86406f020ba0aeb857e62d02337188a (MD5) / As medidas de controle de parasitas de ruminantes são baseadas quase que exclusivamente no uso de antiparasitários comerciais, provocando o rápido desenvolvimento da resistência. O objetivo deste estudo foi verificar o efeito carrapaticida, in vitro, sobre Rhipicephalus (Boophilus) microplus de bovinos, e o efeito anti-helmíntico, in vitro e in vivo, sobre Haemonchus contortus de ovinos. Nos testes realizados sobre R. (B.) microplus, o extrato aquoso de abacaxi e a bromelina foram avaliados em oito concentrações. Fêmeas ingurgitadas foram imersas nos tratamentos e acondicionadas em incubadora para observação da sobrevivência, postura e eclodibilidade das larvas. Para o teste com larvas, cerca de 100 larvas foram colocadas em papéis-filtro impregnados, incubados e as leituras realizadas após 24 horas. O ensaio in vitro foi baseado nos testes de eclodibilidade dos ovos (TEO) e no de desenvolvimento larvar (TDL). No teste in vivo, 36 ovinos infectados artificialmente com H. contortus foram divididos em seis grupos (n=6). No G1 foi administrado 2 g/kg PV do EACA durante três dias, no G2 - 2 g/kg PV do resíduo industrial do abacaxi durante 60 dias, no G3 – 180 mg/animal da bromelina em dose única, G4 – controle negativo I (dos grupos G1 e G2), G5 – controle positivo (10 mg/kg PV de fosfato de levamisol) e G6 – controle negativo II (do grupo G3). Foram realizadas contagens de ovos por grama de fezes (OPG) até o dia 28 após o tratamento. Os resultados do estudo do abacaxi sobre o carrapato demonstraram que, no teste de fêmeas ingurgitadas, as concentrações mais efetivas foram 125, 250 e 500 mg/mL: 33, 48 e 59% para o EACA e 27, 51 e 55%, para a bromelina. As CL50 e CL90 foram, respectivamente, as seguintes: 276 e 8691 mg/mL para o EACA, e 373 e 5172 mg/mL para a bromelina. Nenhuma das diluições estudadas apresentou eficácia contra... / The measures to control parasites in ruminants are almost exclusively based on use of commercial chemical compounds, causing rapid development of resistance. The objective of this study was to investigate the acaricidal effect, in vitro, on Rhipicephalus (Boophilus) microplus in cattle and the anthelmintic effect, in vitro and in vivo, on Haemonchus contortus in sheep. In the tests on R. (B.) microplus, an aqueous extract of pineapple and bromelain were evaluated at eight concentrations. Engorged females were dipped and placed in a incubator for observation of survival, oviposition and egg hatching. For the larvae, about 100 larvae were placed on filter papers impregnated, incubated and readings were taken after 24 hours. The in vitro experiment used the egg hatch test (EHT) and larval development test (LDT). In the in vivo test, 36 sheep artificially infected with H. contortus were divided into six groups (n=6). In the G1 – 2 g/kg BW of AEPS for three days; G2 – 2 g/kg BW of the industrial pineapple residue during 60 days.; G3 – 180 mg/animal of bromelain in a single dose, G4 – negative control I (of groups G1 and G2), G5 – positive control (10 mg/kg BW of levamisole phosphate); and G6 – negative control II (of group G3). The eggs per gram (EPG) of feces were counted until the day 28 after treatment. In the test with engorged cattle ticks, the greatest efficacies were at dilutions of 125, 250 and 500 mg/mL: 33, 48 and 59% for the AEPS and 27, 51 and 55% for bromelain. The LC50 and LC90 were respectively as follows: 276 and 8691 mg/mL for the AEPS, and 373 and 5172 mg/mL for the bromelain. None of the dilutions studied was effective against R. (B.) microplus larvae. In the experiment with the nematode H. contortus, the LC50 and LC90 in the EHT were respectively 31 and 81 mg/mL for the AEPS and 0.50 and 2 mg/mL for the bromelain... (Complete abstract click electronic access below)

Made available in DSpace on 2015-01-26T13:21:22Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-02-21Bitstream added on 2015-01-26T13:30:37Z : No. of bitstreams: 1 000797427.pdf: 235775 bytes, checksum: 6170518ccd58094fe5f34d579507751d (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Objetivo deste trabalho foi avaliar o efeito da dieta e da imunidade sobre o estabelecimento de Haemonchus contortus em cordeiros. O experimento foi delineado em fatorial 2 x 2, duas dietas (feno ou feno+concentrado) e dois estados imunológicos (suprimido ou normal). Os animais foram distribuídos aleatoriamente em quatro grupos experimentais (n=7): feno suprimido, feno normal, feno+concentrado suprimido e feno+concentrado normal. O feno utilizado foi de Cynodon dactylon (CV coast cross) a vontade e a ração ofertada em quantidade equivalente a 3 % do peso, com reajustes semanais. A imunossupressão foi realizada 4 horas antes da infecção artificial e repetida semanalmente, foi utilizado Metilprednisolona na dose de 1.33 mg/kg de peso vivo (intramuscular). O período de adaptação a dieta foi de 22 dias. Cada cordeiro recebeu 4000 larvas infectantes de H. contortus. Foram avaliados os padrões hematológicos, desempenho, resposta imunológica semanalmente e realizado contagens de ovos por grama de fezes (OPG) no 21º, 24º e 27º dias após infecção. Aos 28 dias pós infecção os animais foram eutanasiados e os parasitas quantificados e mensurado seu comprimento e quantidade de ovos em seu interior. As médias foram comparadas por análise de variância a 5% de significância. A carga parasitária foi semelhante em ambos os grupos, no entanto o OPG foi inferior em todas as coletas nos animais que recebiam feno+concentrado (P < 0,05). O imunossupressor foi eficiente em reduzir a contagem de IgG plasmático e mastócitos e eosinófilos na mucosa do abomaso, no entanto houve pouco favorecimento ao parasita frente a imunossupressão, que favoreceu apenas no tamanho dos parasitas machos (P < 0,05). Maior ganho em peso diário ocorreu nos animais que recebiam concentrado, no entanto os do grupo feno+concentrado normal apresentaram maiores ganhos que o feno+concentrado suprimido (P < 0,05). A alimentação com concentrado não exerce efeito ... / The aim of this study was evaluate the effect of diet and immunity on the establishment of Haemonchus contortus in lambs. The trial was in a 2 x 2 factorial design, using two diets (hay or hay+concentrate) and two immune states (normal or supressed). The animals were randomly in four experimental groups (n = 7): hay suppressed, hay normal, hay+concentrate suppressed and hay+concentrate normal. The hay was of Cynodon dactylon (CV Coast cross) and will concentrate offered equivalent to 3% by weight, with weekly adjustments amount. Immunosuppression was performed four hours before artificial infection and repeated weekly, Methylprednisolone was used at a dose 1.33 mg/kg bodyweight (intramuscular) The adaptation period to the diet was 22 days. Each lamb received 4,000 infective larvae (L3) of H. contortus. Haematological, performance, immune response weekly and fecal egg count (FEC) at 21, 24 and 27 days after infection were evaluate. The animals were slaughter 28 days after infection, and the parasites recovered were quantified and measured the length and number of eggs in females were counts. Means were compare by analysis of variance at 5 % significance (Tukey´s test). The worm burden was similar in both groups, however the FEC was lower in all samples on animals that received hay+concentrate (P < 0.05). The immunosuppressive was effective on reduce the IgG plasma levels and number of abomasal mucosal mast cell and eosinophils, however there was little favouritism to parasite against immunosuppression, which favoured only in the size of male parasites ( P < 0.05). The greater daily weight gain was observed in animals receiving concentrate, however the group hay+concentrate normal showed higher weight gains than hay+concentrate suppressed group (P < 0.05). The supply of concentrate does not exercise direct effect on establishment of H. contortus. Supplementation favours weight gain, lessens the pathogenic effects of the parasite and leads ...

Orientador: Gervásio Henrique Bechara / Coorientador: Gilson Pereira de Oliveira / Banca: Estevam Guilherme Lux Hoppe / Banca: Gislaine Cristina Roma / Banca: Patrícia Rosa de Oliveira / Banca: Ana Carolina de Souza Chagas / Resumo: As medidas de controle de parasitas de ruminantes são baseadas quase que exclusivamente no uso de antiparasitários comerciais, provocando o rápido desenvolvimento da resistência. O objetivo deste estudo foi verificar o efeito carrapaticida, in vitro, sobre Rhipicephalus (Boophilus) microplus de bovinos, e o efeito anti-helmíntico, in vitro e in vivo, sobre Haemonchus contortus de ovinos. Nos testes realizados sobre R. (B.) microplus, o extrato aquoso de abacaxi e a bromelina foram avaliados em oito concentrações. Fêmeas ingurgitadas foram imersas nos tratamentos e acondicionadas em incubadora para observação da sobrevivência, postura e eclodibilidade das larvas. Para o teste com larvas, cerca de 100 larvas foram colocadas em papéis-filtro impregnados, incubados e as leituras realizadas após 24 horas. O ensaio in vitro foi baseado nos testes de eclodibilidade dos ovos (TEO) e no de desenvolvimento larvar (TDL). No teste in vivo, 36 ovinos infectados artificialmente com H. contortus foram divididos em seis grupos (n=6). No G1 foi administrado 2 g/kg PV do EACA durante três dias, no G2 - 2 g/kg PV do resíduo industrial do abacaxi durante 60 dias, no G3 - 180 mg/animal da bromelina em dose única, G4 - controle negativo I (dos grupos G1 e G2), G5 - controle positivo (10 mg/kg PV de fosfato de levamisol) e G6 - controle negativo II (do grupo G3). Foram realizadas contagens de ovos por grama de fezes (OPG) até o dia 28 após o tratamento. Os resultados do estudo do abacaxi sobre o carrapato demonstraram que, no teste de fêmeas ingurgitadas, as concentrações mais efetivas foram 125, 250 e 500 mg/mL: 33, 48 e 59% para o EACA e 27, 51 e 55%, para a bromelina. As CL50 e CL90 foram, respectivamente, as seguintes: 276 e 8691 mg/mL para o EACA, e 373 e 5172 mg/mL para a bromelina. Nenhuma das diluições estudadas apresentou eficácia contra... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The measures to control parasites in ruminants are almost exclusively based on use of commercial chemical compounds, causing rapid development of resistance. The objective of this study was to investigate the acaricidal effect, in vitro, on Rhipicephalus (Boophilus) microplus in cattle and the anthelmintic effect, in vitro and in vivo, on Haemonchus contortus in sheep. In the tests on R. (B.) microplus, an aqueous extract of pineapple and bromelain were evaluated at eight concentrations. Engorged females were dipped and placed in a incubator for observation of survival, oviposition and egg hatching. For the larvae, about 100 larvae were placed on filter papers impregnated, incubated and readings were taken after 24 hours. The in vitro experiment used the egg hatch test (EHT) and larval development test (LDT). In the in vivo test, 36 sheep artificially infected with H. contortus were divided into six groups (n=6). In the G1 - 2 g/kg BW of AEPS for three days; G2 - 2 g/kg BW of the industrial pineapple residue during 60 days.; G3 - 180 mg/animal of bromelain in a single dose, G4 - negative control I (of groups G1 and G2), G5 - positive control (10 mg/kg BW of levamisole phosphate); and G6 - negative control II (of group G3). The eggs per gram (EPG) of feces were counted until the day 28 after treatment. In the test with engorged cattle ticks, the greatest efficacies were at dilutions of 125, 250 and 500 mg/mL: 33, 48 and 59% for the AEPS and 27, 51 and 55% for bromelain. The LC50 and LC90 were respectively as follows: 276 and 8691 mg/mL for the AEPS, and 373 and 5172 mg/mL for the bromelain. None of the dilutions studied was effective against R. (B.) microplus larvae. In the experiment with the nematode H. contortus, the LC50 and LC90 in the EHT were respectively 31 and 81 mg/mL for the AEPS and 0.50 and 2 mg/mL for the bromelain... (Complete abstract click electronic access below) / Doutor

Resumo: Avaliou-se o efeito do extrato de própolis na redução da verminose e sobre os parâmetros hematológicos e bioquímicos de 18 cordeiros Ile de France (nove machos não-castrados e nove fêmeas) dos 5 aos 15 kg de peso corporal, no desmame. Os animais foram distribuídos em três tratamentos, constituídos por dietas isoprotéicas e isoenergéticas, com adição ao concentrado de 0, 15 e 30 mg de extrato de própolis a 11%/kg de peso corporal . O delineamento experimental foi o inteiramente casualizado em esquema de parcela subdividida no tempo. Observou -se que a contagem de ovos por grama de fezes foi influenciada pelos diferentes tratament os. Nos cordeiros estudados, a ad ição de 30 mg de extrato de própolis foi mais efetiva em reduzir o número de ovos tipo Stron gylida por grama de fezes, do que a de 15 mg e a que não continha o extrato, indicando uma possível redução na ovoposição dos endoparasitos. As contagens globais de hemácias, neutrófilos bastonetes e monócitos, taxa de hemoglobina e o percentual de hematócrito apresentaram efeito para interação entre os tratamentos e as datas de observação. Não houve diferenças para as contagens de leucócitos, neutrófilos segmentados e linfócitos, já os eosinófilos variaram dentro do período de coleta. Em relação aos parâmetros bioquímicos analisados, o extrato de própolis não afetou as concentrações séricas de uréia, creatinina, albumina, bilirrubina direta e indireta, e plasmática de glicose. Já os níveis séricos d e colesterol, proteínas totais e bilirrubina total diferiram entre os tratamentos. Com base nos resultados, concluí-se que a adição de extrato de própolis às rações dos cordeiros não provocou alterações importantes nos parâmetros hematológicos e bioquímicos, que indicassem reações adversas à sua administração, além de ser uma possível alternativa no controle da verminose em cordeiros. / Abstract: The effect of the propolis extract was evaluated in the reduction of the nematodes and on the hematological and bioch emical parameters of 18 lambs Ile of France (nine no-castrated males and nine females) of the 5 to the 15 kg of corporal weight, in it weans them. The animals were distributed in three treatments, constituted by protein and energy concentration diets, with addition to the concentrate of 0, 15 and 30 mg of propolis extract to 11%/kg of corporal weight. The experimental delineation was entirely occurred in portion outline subdivided in the time. It was observed that the counting of eggs for gram of feces was influenced by the different treatments. In the studied lambs, the addition of 30 mg of extract of propolis was more effective in reducing the number of eggs type Strongylida for gram of feces, than the one of 15 mg and the one that didn't contain the extra ct, indicating a possible reduction in the eggs of the endoparasites. The global countings of the red cells, neutrophiles and monocytes, hemoglobin tax and the percentile of hematocrits presented effect for interaction among the treatments and the dates fo r observation. There were not differences for the leucocytes countings, segmented neutrophiles and lymphocytes, whereas the eosinophiles varied inside of the collection period. In relation to the analyzed biochemical parameters, the extract of propolis did n't affect the serum urea, creatinine, albumin, direct and indirect, and plasmatic bilirubin concentration of glucose. The serum levels of cholesterol, total proteins and total bilirubin differed among the treatments. On base in the results, I was ended th at the addition of extract of propolis to the rations of the lambs didn't provoke important alterations in the hematological and biochemical parameters, that they indicated adverse reactions to his/her administration, besides being a possible alternative in the control of the nematodes in lambs. / Orientador: Americo Garcia da Silva Sobrinho / Coorientador: Áureo Evangelista Santana / Banca: Adjair Antonio do Nascimento / Banca: Lucimar Pontara Peres de Moura / Mestre

Orientador: Alessandro Francisco Talamini do Amarante / Banca: Alexandre Secorun Borges / Banca: Helder Louvandini / Resumo: Objetivo deste trabalho foi avaliar o efeito da dieta e da imunidade sobre o estabelecimento de Haemonchus contortus em cordeiros. O experimento foi delineado em fatorial 2 x 2, duas dietas (feno ou feno+concentrado) e dois estados imunológicos (suprimido ou normal). Os animais foram distribuídos aleatoriamente em quatro grupos experimentais (n=7): feno suprimido, feno normal, feno+concentrado suprimido e feno+concentrado normal. O feno utilizado foi de Cynodon dactylon (CV coast cross) a vontade e a ração ofertada em quantidade equivalente a 3 % do peso, com reajustes semanais. A imunossupressão foi realizada 4 horas antes da infecção artificial e repetida semanalmente, foi utilizado Metilprednisolona na dose de 1.33 mg/kg de peso vivo (intramuscular). O período de adaptação a dieta foi de 22 dias. Cada cordeiro recebeu 4000 larvas infectantes de H. contortus. Foram avaliados os padrões hematológicos, desempenho, resposta imunológica semanalmente e realizado contagens de ovos por grama de fezes (OPG) no 21º, 24º e 27º dias após infecção. Aos 28 dias pós infecção os animais foram eutanasiados e os parasitas quantificados e mensurado seu comprimento e quantidade de ovos em seu interior. As médias foram comparadas por análise de variância a 5% de significância. A carga parasitária foi semelhante em ambos os grupos, no entanto o OPG foi inferior em todas as coletas nos animais que recebiam feno+concentrado (P < 0,05). O imunossupressor foi eficiente em reduzir a contagem de IgG plasmático e mastócitos e eosinófilos na mucosa do abomaso, no entanto houve pouco favorecimento ao parasita frente a imunossupressão, que favoreceu apenas no tamanho dos parasitas machos (P < 0,05). Maior ganho em peso diário ocorreu nos animais que recebiam concentrado, no entanto os do grupo feno+concentrado normal apresentaram maiores ganhos que o feno+concentrado suprimido (P < 0,05). A alimentação com concentrado não exerce efeito ... / Abstract: The aim of this study was evaluate the effect of diet and immunity on the establishment of Haemonchus contortus in lambs. The trial was in a 2 x 2 factorial design, using two diets (hay or hay+concentrate) and two immune states (normal or supressed). The animals were randomly in four experimental groups (n = 7): hay suppressed, hay normal, hay+concentrate suppressed and hay+concentrate normal. The hay was of Cynodon dactylon (CV Coast cross) and will concentrate offered equivalent to 3% by weight, with weekly adjustments amount. Immunosuppression was performed four hours before artificial infection and repeated weekly, Methylprednisolone was used at a dose 1.33 mg/kg bodyweight (intramuscular) The adaptation period to the diet was 22 days. Each lamb received 4,000 infective larvae (L3) of H. contortus. Haematological, performance, immune response weekly and fecal egg count (FEC) at 21, 24 and 27 days after infection were evaluate. The animals were slaughter 28 days after infection, and the parasites recovered were quantified and measured the length and number of eggs in females were counts. Means were compare by analysis of variance at 5 % significance (Tukey's test). The worm burden was similar in both groups, however the FEC was lower in all samples on animals that received hay+concentrate (P < 0.05). The immunosuppressive was effective on reduce the IgG plasma levels and number of abomasal mucosal mast cell and eosinophils, however there was little favouritism to parasite against immunosuppression, which favoured only in the size of male parasites ( P < 0.05). The greater daily weight gain was observed in animals receiving concentrate, however the group hay+concentrate normal showed higher weight gains than hay+concentrate suppressed group (P < 0.05). The supply of concentrate does not exercise direct effect on establishment of H. contortus. Supplementation favours weight gain, lessens the pathogenic effects of the parasite and leads ... / Mestre

Submitted by Mario BC (mario@bc.ufrpe.br) on 2018-02-27T12:53:13Z No. of bitstreams: 1 Rodolfo Luiz Godoy do Amaral.pdf: 2067398 bytes, checksum: e01b4173ff3a3945dcc3b8f34170148b (MD5) / Made available in DSpace on 2018-02-27T12:53:13Z (GMT). No. of bitstreams: 1 Rodolfo Luiz Godoy do Amaral.pdf: 2067398 bytes, checksum: e01b4173ff3a3945dcc3b8f34170148b (MD5) Previous issue date: 2016-02-29 / The sheep and goat farming suffer great economic losses due to parasitism by gastrointestinal nematodes, reaching derail this exploration in some regions. The control of such parasites is done primarily with the use of anthelmintic drugs. The inefficiency of this type of control are the first signs of anthelmintic resistance appearance (AR), which constitutes one of the main negative and limiting factors for animal production, since it prevents the effective control of helminths in small ruminants, with adverse consequences on production rates. The anthelmintic resistance in nematodes of small ruminants for the three groups most commonly used drugs: benzimidazole (BZs) imidazotiazóis (LEV) and macrocyclic lactones (MLs) has been reported and have grown rapidly in different regions of the world. The efficient control of these parasites and the early diagnosis of anthelmintic resistance, especially in Haemonchus contortus should be recommended in order to economically viable sheep and goat breeding. Knowledge of various genetic aspects of this phenomenon can increase the shelf life of drugs currently used, and therefore try to preserve the susceptibility of parasites. This work has the objective of diagnosing anthelmintic resistance in properties of sheep and goats and to analyze the relationship with the management of the properties and identify the presence of DNA responsible for anthelmintic resistance in helminth populations of small ruminants in the state of Pernambuco. The methodological procedures were approved by the Ethics Committee on Animal Use - CEUA - UFRPE. Samples from adult specimens of H. contortus were obtained from animals naturally infected with gastrointestinal helminths belonging to properties located in the cities of the state of Pernambuco: Sairé (Agreste), Recife (Metropolitan Region), Bonito (Agreste), Moreno (Metropolitan Region) Gameleira (South Forest), Camocim de São Félix (Agreste), Serra Talhada (Sertão). The properties of Recife and Bonito created only goats and the others only sheep. For PCR, samples from adult specimens of H. contortus were obtained from animals naturally infected with gastrointestinal helminths, which were stored in Eppendorf tubes with PBS at 4 ° C, identified and sexed in Parasitic Diseases Laboratory of Domestic Animals - UFRPE. It was performed a pool of 20 adult males of H. contortus from each property for the extraction of DNA. The extracted DNA samples were analyzed in Parasitic Diseases Laboratory at the Federal University of Parana. PCR in order to amplify gene fragments encoding 1 isotype of β-tubulin which held the sequence of codon 200 was performed;it was also performing a PCR to identify the species of helminths collected. Both amplification reactions were performed in a thermocycler Veriti Life gradient. A sample of 20 uL of each reaction was analyzed on a 0.8% agarose gel and visualized under UV light. PCR products were purified with the commercial PureLink® Quick Gel Extraction Kit (Invitrogen, USA) and sequenced by capillary electrophoresis (Sanger method) using the ABI3130 platform (Life Technologies, USA). Anthelmintic efficacy by "in vivo" method was also evaluated the by means of Fecal Egg Counting Reduction Test (FECRT) using the formula:% efficacy = [(mean OPGpre-treatment - average OPGpost-treatment) / average OPGpre-treatment)] x 100. It was detected DNA responsible for anthelmintic benzimidazole resistance (BZ) in all the evaluated properties. The percentage of anthelmintic efficacy for moxidectin presented as effective only seven days after treatment. Several management failures of control of gastrointestinal helminths were found. The results obtained confirm the anthelmintic benzimidazole resistance in H. contortus goats and sheep in the state of Pernambuco - Brazil, becoming the first record of molecular diagnosis of RA in ruminants in the State. also characterize the decline of the anthelmintic efficacy of moxidectin in herds. Both situations are favored by inadequate management practices and indiscriminate use of chemicals for the control of gastrointestinal helminths. / A caprinovinocultura sofre grande prejuízo econômico devido ao parasitismo por nematódeos gastrintestinais, chegando a inviabilizar esta exploração em algumas regiões. O controle desse parasitismo é feito basicamente com a utilização de drogas anti-helmínticas. A ineficiência neste tipo de controle são os primeiros sinais do aparecimento de resistência anti-helmíntica (RA), que se constitui em um dos principais fatores negativos e limitantes para a produção animal, uma vez que inviabiliza o controle efetivo da helmintose dos pequenos ruminantes, com reflexos desfavoráveis nos índices produtivos. A resistência anti-helmíntica em nematódeos de pequenos ruminantes para os três grupos de drogas mais comumente utilizados: benzimidazóis (BZs), imidazotiazóis (LEV) e lactonas macrocíclicas (LMs) vem sendo relatada e têm crescido rapidamente em diferentes regiões do mundo. O controle eficiente destes parasitos e o diagnóstico precoce da resistência anti-helmíntica, especialmente em Haemonchus contortus, devem ser preconizados a fim de viabilizar economicamente a criação de ovinos e caprinos. O conhecimento dos vários aspectos genéticos deste fenômeno poderá aumentar a vida útil dos fármacos atualmente utilizados, e consequentemente tentar preservar a susceptibilidade dos parasitos. Neste trabalho tem-se como objetivo diagnosticar a resistência anti-helmíntica em propriedades de criação de ovinos e caprinos e analisar a relação com o manejo instituído nas propriedades e identificar a presença do DNA responsável pela resistência anti-helmíntica em populações de helmintos de pequenos ruminantes no estado de Pernambuco. Os procedimentos metodológicos foram aprovados pela Comissão de Ética no Uso de Animais – CEUA – UFRPE. Amostras de espécimes adultos de H. contortus foram obtidas de animais naturalmente parasitados por helmintos gastrintestinais pertencentes a propriedades localizadas nas cidades do estado de Pernambuco: Sairé (Agreste), Recife (Região Metropolitana), Bonito (Agreste), Moreno (Região Metropolitana), Gameleira (Mata Sul), Camocim de São Félix (Agreste), Serra Talhada (Sertão). Nas propriedades de Recife e Bonito criavam-se apenas caprinos e nas demais apenas ovinos. Para realização da PCR, amostras de espécimes adultos de H. contortus foram obtidas de animais naturalmente parasitados por helmintos gastrintestinais, os quais foram estocados em tubos de eppendorf com PBS a 4ºC, identificados e sexados no Laboratório de Doenças Parasitárias dos Animais Domésticos – UFRPE. Formou-se um pool de 20 indivíduos machos adultos de H. contortus de cada propriedade, dos quais procedeu-se à extração de DNA. As amostras de DNA extraídas foram analisadas no Laboratório de Doenças Parasitárias da Universidade Federal do Paraná. Foi realizada a PCR com o objetivo amplificar fragmentos de genes que codificam o isotipo 1 da β- tubulina que possuíam as sequências de códon 200, realizando-se também uma PCR para identificação das espécies dos helmintos coletados. As duas reações de amplificação foram conduzidas em termociclador Veriti Life com gradiente. Uma amostra de 20 μl de cada reação foi analisada sobre um gel de agarose 0,8 % e visualizada sob luz UV. Os produtos de PCR foram purificados com o kit comercial PureLink® Quick Gel Extraction (Invitrogen, USA) e sequenciados por eletroforese capilar (Método de Sanger) utilizando a plataforma ABI3130 (Life Technologies, USA). Avaliou-se também a eficácia anti-helmíntica por método “in vivo” por meio do teste de redução do número de ovos por grama de fezes (TRCOF), usando a fórmula: % eficácia = [(média de OPGpré-tratamento - média de OPGpós-tratamento) / média de OPGprétratamento )] X 100. Detectou-se DNA responsável pela resistência anti-helmíntica aos benzimidazóis (BZ) em todas as propriedades analisadas. Os percentuais de eficácia anti-helmíntica para a moxidectina apresentaram-se como efetivos apenas aos sete dias pós-tratamento. Diversas falhas de manejo do controle das helmintoses gastrintestinais foram encontradas. Os resultados obtidos confirmam a resistência anti-helmíntica aos benzimidazóis em H. contortus de caprinos e ovinos no estado de Pernambuco, constituindo-se no primeiro registro de diagnóstico molecular da RA em ruminantes no Estado. Caracterizam também o declínio da efetividade anti-helmíntica da moxidectina nos rebanhos. Ambas as situações são favorecida por práticas de manejo inadequadas e uso indiscriminado de produtos químicos para o controle de helmintos gastrintestinais.

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

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

No description available.

Made available in DSpace on 2014-06-11T19:32:52Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-02-08Bitstream added on 2014-06-13T19:22:39Z : No. of bitstreams: 1 hoppe_egl_dr_jabo.pdf: 1540256 bytes, checksum: 5f7596806464f080af8ea17e52f8fafd (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / A adequada compreensão dos mecanismos fisiopatológicos das doenças que afetam os animais silvestres contribui positivamente para a melhoria das condições de manutenção desses animais em cativeiro e fornece subsídios para a elaboração de planos de conservação de animais de vida livre. Diversos relatos de Haemonchus contortus em cervídeos de cativeiro e vida livre, bem como a importância desse nematódeo para a ovinocultura, motivaram o presente estudo. Inicialmente, foram determinados valores de normalidade para os parâmetros hematológicos e bioquímicos, bem como testada possível influência do sexo sobre eles. Num segundo momento, os animais foram separados em grupos e infectados experimentalmente. O processo infeccioso foi monitorado por um período de 60 dias, ao final do qual os animais foram submetidos à eutanásia e necropsiados. O processo infeccioso estabelecido, em intensidade parasitária similar à observada em animais de vida livre, não foi capaz de suscitar alterações hematológicas, bioquímicas ou anátomo-patológicas relevantes, tampouco interferir na ingestão de alimentos, sugerindo que, apesar da possibilidade de parasitismo, a infecção por H. contortus, nos níveis deste experimento, não provocam doença clínica nos animais. À luz dos dados morfométricos e parasitológicos, sugere-se uma má adaptação do H. contortus aos veados-catingueiros, apesar da possibilidade de manutenção do parasita nestes animais. / The comprehension of the pathophysiological aspects concerning wild animals’ diseases contributes favourably to the improvement of captive rearing of these animals, and also to the elaboration of conservation action plans for free-ranging animals. The diverse reports on deer parasitism by Haemonchus contortus and the importance of this nematode for sheep production motivated this research. First, reference values for haematological and biochemical parameters were determined for this species and the gender influence on them was tested. Then, the animals were divided in groups and infected. The infectious process was monitored over a 60 days period, and then the animals were killed and necropsied. The established infection, similar to natural acquired infections, was insufficient to lead to haematological, biochemical or pathological changes or interfere with food uptake. Based on this, despite the possibility of infection, H. contortus did not cause clinical disease in gray-brocket deer. Moreover, the morphometric and parasitological data suggest a maladaptation of H. contortus in this host species.