Biomarcadores sanguíneos de exposição à zearalenona e aplicabilidade na avaliação da eficiência de aditivos antimicotoxinas em suínos e bovinos
Autor(a) principal: | |
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Data de Publicação: | 2020 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | Manancial - Repositório Digital da UFSM |
Texto Completo: | http://repositorio.ufsm.br/handle/1/22220 |
Resumo: | The inclusion of antimycotoxins additives (AMA) in the diet of production animals has been used to avoid mycotoxins exposure. To confirm the efficacy of such products in vivo, measurement of mycotoxins and/or their metabolites in biological fluids is preconized. This work consists of two studies and aimed to: (a) improve an analytical methodology based on High Performance Liquid Chromatography Coupled to Tandem Mass Spectrometry (HPLC-MS/MS) to detect and quantify zearalenone (ZEA) and its metabolites, α-zearalenol (α-ZOL), β-zearalenol, α-zearalanol, β-zearalanol (β-ZAL) and zearalanone, in blood serum; and (b) to conduct tests in species which are susceptible to ZEA to evaluate exposure to mycotoxicosis and to use serological biomarkers to assess the efficacy of AMA. In the first study, 24 beef heifers were randomly assigned to receive one of these treatments (n=6/treatament): T1) basal diet (control); T2) basal diet + 5 mg/kg ZEA; T3) basal diet + 5 mg/kg ZEA + 2.5 kg/t AMA; and T4) basal diet + 5 mg/kg ZEA + 5.0 kg/t AMA. The trial lasted 37 days. Blood was collected on different days after the diet was given, and the samples were centrifuged to obtain the blood serum and then analysed by HPLC-MS/MS. Among the analyzed metabolliates, β-ZAL was detected above the limit of quantification both in the unconjugated (>0.60 μg/kg) and conjugated (>1.70 μg/kg) forms. The remaining metabolites presented concentrations under the limit of detection. In the efficacy evaluation of the AMA, there was no significant difference (P>0.05) between the treatments with and without AMA at the tested levels of inclusion. In the second study, 70 pre-pubertal gilts were distributed in seven feeding groups (n=10/treatment) receiving diets with two levels of three clay-based AMAs (0.25% and 0.50%) and three levels of ZEA (0, 0.75 and 1 mg/kg) for 42 days. Initial and final body weight (BW), mean daily weight gain, mean daily feed intake, feed conversion and vulva volume were assessed at the time of blood sampling to determine ZEA and its metabolites in the serum. Blood samples were spun to obtain the serum, which was analyzed via HPLC-MS/MS. When the experimental period ceased, the animals were slaughtered and eviscerated in order to evaluate weight and length of the reproductive tract. The zootechnical performance was not affected by the presence of ZEA in the diet, and inclusion of AMA did not determine differences (P>0.05). ZEA caused a significant increase (P<0.05) in BW, length of the reproductive tract and vulva measurement (width, length and area), with no difference (P>0.05) between treatments with the addition of AMA to the diet (P>0.05). With respect to the evaluated metabolites, α-ZOL was detected above the limit of quantification in both unconjugated (>0.15 μg/kg) and unconjugated (>1.58 μg/kg) forms. The concentration of the remaining metabolites was below the limit of detection, thus confirming the results of the efficacy of AMA (P<0.05). The findings in this study contribute to the search of quality of the marketed AMA as well as to the assessment of analytical methods and biomarkers; these substances may represent an alternative to confirm the efficacy of such products in heifers and gilts. |
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2021-09-15T10:49:56Z2021-09-15T10:49:56Z2020-02-14http://repositorio.ufsm.br/handle/1/22220The inclusion of antimycotoxins additives (AMA) in the diet of production animals has been used to avoid mycotoxins exposure. To confirm the efficacy of such products in vivo, measurement of mycotoxins and/or their metabolites in biological fluids is preconized. This work consists of two studies and aimed to: (a) improve an analytical methodology based on High Performance Liquid Chromatography Coupled to Tandem Mass Spectrometry (HPLC-MS/MS) to detect and quantify zearalenone (ZEA) and its metabolites, α-zearalenol (α-ZOL), β-zearalenol, α-zearalanol, β-zearalanol (β-ZAL) and zearalanone, in blood serum; and (b) to conduct tests in species which are susceptible to ZEA to evaluate exposure to mycotoxicosis and to use serological biomarkers to assess the efficacy of AMA. In the first study, 24 beef heifers were randomly assigned to receive one of these treatments (n=6/treatament): T1) basal diet (control); T2) basal diet + 5 mg/kg ZEA; T3) basal diet + 5 mg/kg ZEA + 2.5 kg/t AMA; and T4) basal diet + 5 mg/kg ZEA + 5.0 kg/t AMA. The trial lasted 37 days. Blood was collected on different days after the diet was given, and the samples were centrifuged to obtain the blood serum and then analysed by HPLC-MS/MS. Among the analyzed metabolliates, β-ZAL was detected above the limit of quantification both in the unconjugated (>0.60 μg/kg) and conjugated (>1.70 μg/kg) forms. The remaining metabolites presented concentrations under the limit of detection. In the efficacy evaluation of the AMA, there was no significant difference (P>0.05) between the treatments with and without AMA at the tested levels of inclusion. In the second study, 70 pre-pubertal gilts were distributed in seven feeding groups (n=10/treatment) receiving diets with two levels of three clay-based AMAs (0.25% and 0.50%) and three levels of ZEA (0, 0.75 and 1 mg/kg) for 42 days. Initial and final body weight (BW), mean daily weight gain, mean daily feed intake, feed conversion and vulva volume were assessed at the time of blood sampling to determine ZEA and its metabolites in the serum. Blood samples were spun to obtain the serum, which was analyzed via HPLC-MS/MS. When the experimental period ceased, the animals were slaughtered and eviscerated in order to evaluate weight and length of the reproductive tract. The zootechnical performance was not affected by the presence of ZEA in the diet, and inclusion of AMA did not determine differences (P>0.05). ZEA caused a significant increase (P<0.05) in BW, length of the reproductive tract and vulva measurement (width, length and area), with no difference (P>0.05) between treatments with the addition of AMA to the diet (P>0.05). With respect to the evaluated metabolites, α-ZOL was detected above the limit of quantification in both unconjugated (>0.15 μg/kg) and unconjugated (>1.58 μg/kg) forms. The concentration of the remaining metabolites was below the limit of detection, thus confirming the results of the efficacy of AMA (P<0.05). The findings in this study contribute to the search of quality of the marketed AMA as well as to the assessment of analytical methods and biomarkers; these substances may represent an alternative to confirm the efficacy of such products in heifers and gilts.A inclusão de aditivos antimicotoxinas (AAM) na dieta de animais de produção tem sido utilizada para evitar a exposição às micotoxinas. Preconiza-se a medição das micotoxinas e/ou seus metabólitos em fluidos biológicos como marcadores para confirmar a eficácia desses produtos. Este trabalho consiste em dois estudos e objetivou: (a) aprimorar uma metodologia analítica baseada em cromatografia líquida de alta eficiência acoplada à espectrometria de massas (HPLC-MS/MS) para a detecção e quantificação de zearalenona (ZEA) e seus metabólitos, α-zearalenol (α-ZEL), β-zearalenol, α-zearalanol, β-zearalanol (β-ZAL) e zearalanona, em soro sanguíneo; e (b) executar testes em espécies susceptíveis à ZEA para avaliar a exposição à micotoxicose e utilizar biomarcadores sanguíneos para verificar a eficácia de AAM. No primeiro estudo, 24 novilhas de corte foram distribuídas aleatoriamente para receber um destes tratamentos (n=6/tratamento): T1) dieta basal (controle); T2) dieta basal + 5 mg/kg ZEA; T3) dieta basal + 5 mg/kg ZEA + 2,5 kg/t AAM; e T4) dieta basal + 5 mg/kg ZEA + 5 kg/t AAM. A investigação durou 37 dias. Após o fornecimento da dieta, amostras de sangue foram colhidas em diferentes dias, centrifugadas para a obtenção do soro sanguíneo e analisadas por HPLC-MS/MS. Entre os metabólitos analisados, β-ZAL foi detectado acima do limite de quantificação nas formas não conjugadas (>0,60 μg/kg) e conjugadas (>1,70 μg/kg). Os demais metabólitos apresentaram concentrações abaixo do limite de detecção. Na avaliação da eficácia do AAM, não houve diferença significativa (P>0,05) entre os tratamentos com e sem aditivo nos níveis de inclusão avaliados. No segundo estudo, 70 leitoas pré-púberes foram distribuídas em sete grupos (n=10), alimentadas com dietas apresentando dois níveis de inclusão de AAM à base de argila (0,25% e 0,50%) e três níveis de inclusão de ZEA (0, 0,75 e 1 mg/kg) por 42 dias. Foram realizadas avaliações de peso inicial e final, ganho médio diário, consumo médio diário, conversão alimentar e vulvometria junto às colheitas de sangue para avaliação de ZEA e metabólitos em soro. As amostras de sangue foram centrifugadas para a separação do soro e analisadas por HPLC-MS/MS. Ao fim do período experimental, os animais foram abatidos e eviscerados para avaliar o peso e o comprimento do trato reprodutivo. O desempenho zootécnico não foi afetado pela adição de ZEA à dieta, não havendo diferença na inclusão de AAM (P>0,05). A ZEA aumentou significativamente (P<0,05) o peso, o comprimento do trato reprodutivo e o volume vulvar (largura, comprimento e área); não houve diferença entre os tratamentos com a adição de AAM à dieta (P>0,05). Quanto aos metabólitos pesquisados, α-ZEL foi detectado acima do limite de quantificação nas formas não conjugadas (>0,15 μg/kg) e conjugadas (>1,58 μg/kg). A concentração dos demais metabólitos ficou abaixo do limite de detecção, confirmando os resultados da eficácia do AAM (P<0,05). Os resultados obtidos com este trabalho contribuem para a busca da qualidade de AAMs comercializados, bem como para a utilização de métodos analíticos e biomarcadores; estes podem representar uma alternativa para a confirmação da eficácia desses produtos em novilhas e leitoas.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESporUniversidade Federal de Santa MariaCentro de Ciências RuraisPrograma de Pós-Graduação em Medicina VeterináriaUFSMBrasilMedicina VeterináriaAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessAditivo antimicotoxinaBiomarcadorAdsorçãoArgilasHPLC-MS/MSAntimycotoxins additiveBiomarkerAdsorptionClaysCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIABiomarcadores sanguíneos de exposição à zearalenona e aplicabilidade na avaliação da eficiência de aditivos antimicotoxinas em suínos e bovinosSerological biomarkers of zearalenone exposure and their aplicability to assess the efficacy of antimycotoxins additives in swine and bovineinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisMallmann, Carlos Augustohttp://lattes.cnpq.br/5193771213666058Antoniazzi, Alfredo QuitesAlmeida, Carlos Alberto Araújo deRossi, Carlos Augusto RigonKrabbe, Everton LuísSá, Luciano Moraeshttp://lattes.cnpq.br/8281811290390141Tonini, Camila500500000007600600600600600600600f8b7bb02-50a2-468a-91b2-0a75e5374e2f4eae5a85-6864-4f4e-a05c-bfa481d5a83307e69e9e-c8e4-460c-91dd-f3b5aa4e560a82c4cdb9-e9b7-4703-83e4-9a8221ec4f737315aaa1-0836-43a9-9e07-e7f1d0da3bcf5f95e913-f613-49d1-879e-91a7ee4322ede1a1fb2a-9f30-4cd6-865a-48f68c3a6cfcreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSMORIGINALTES_PPGMV_2020_TONINI_CAMILA.pdfTES_PPGMV_2020_TONINI_CAMILA.pdfTese de Doutoradoapplication/pdf1659983http://repositorio.ufsm.br/bitstream/1/22220/1/TES_PPGMV_2020_TONINI_CAMILA.pdf483d49fd15660fe8127e60b040c79c34MD51CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; 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dc.title.por.fl_str_mv |
Biomarcadores sanguíneos de exposição à zearalenona e aplicabilidade na avaliação da eficiência de aditivos antimicotoxinas em suínos e bovinos |
dc.title.alternative.eng.fl_str_mv |
Serological biomarkers of zearalenone exposure and their aplicability to assess the efficacy of antimycotoxins additives in swine and bovine |
title |
Biomarcadores sanguíneos de exposição à zearalenona e aplicabilidade na avaliação da eficiência de aditivos antimicotoxinas em suínos e bovinos |
spellingShingle |
Biomarcadores sanguíneos de exposição à zearalenona e aplicabilidade na avaliação da eficiência de aditivos antimicotoxinas em suínos e bovinos Tonini, Camila Aditivo antimicotoxina Biomarcador Adsorção Argilas HPLC-MS/MS Antimycotoxins additive Biomarker Adsorption Clays CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
title_short |
Biomarcadores sanguíneos de exposição à zearalenona e aplicabilidade na avaliação da eficiência de aditivos antimicotoxinas em suínos e bovinos |
title_full |
Biomarcadores sanguíneos de exposição à zearalenona e aplicabilidade na avaliação da eficiência de aditivos antimicotoxinas em suínos e bovinos |
title_fullStr |
Biomarcadores sanguíneos de exposição à zearalenona e aplicabilidade na avaliação da eficiência de aditivos antimicotoxinas em suínos e bovinos |
title_full_unstemmed |
Biomarcadores sanguíneos de exposição à zearalenona e aplicabilidade na avaliação da eficiência de aditivos antimicotoxinas em suínos e bovinos |
title_sort |
Biomarcadores sanguíneos de exposição à zearalenona e aplicabilidade na avaliação da eficiência de aditivos antimicotoxinas em suínos e bovinos |
author |
Tonini, Camila |
author_facet |
Tonini, Camila |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Mallmann, Carlos Augusto |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/5193771213666058 |
dc.contributor.referee1.fl_str_mv |
Antoniazzi, Alfredo Quites |
dc.contributor.referee2.fl_str_mv |
Almeida, Carlos Alberto Araújo de |
dc.contributor.referee3.fl_str_mv |
Rossi, Carlos Augusto Rigon |
dc.contributor.referee4.fl_str_mv |
Krabbe, Everton Luís |
dc.contributor.referee5.fl_str_mv |
Sá, Luciano Moraes |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/8281811290390141 |
dc.contributor.author.fl_str_mv |
Tonini, Camila |
contributor_str_mv |
Mallmann, Carlos Augusto Antoniazzi, Alfredo Quites Almeida, Carlos Alberto Araújo de Rossi, Carlos Augusto Rigon Krabbe, Everton Luís Sá, Luciano Moraes |
dc.subject.por.fl_str_mv |
Aditivo antimicotoxina Biomarcador Adsorção Argilas HPLC-MS/MS |
topic |
Aditivo antimicotoxina Biomarcador Adsorção Argilas HPLC-MS/MS Antimycotoxins additive Biomarker Adsorption Clays CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
dc.subject.eng.fl_str_mv |
Antimycotoxins additive Biomarker Adsorption Clays |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
description |
The inclusion of antimycotoxins additives (AMA) in the diet of production animals has been used to avoid mycotoxins exposure. To confirm the efficacy of such products in vivo, measurement of mycotoxins and/or their metabolites in biological fluids is preconized. This work consists of two studies and aimed to: (a) improve an analytical methodology based on High Performance Liquid Chromatography Coupled to Tandem Mass Spectrometry (HPLC-MS/MS) to detect and quantify zearalenone (ZEA) and its metabolites, α-zearalenol (α-ZOL), β-zearalenol, α-zearalanol, β-zearalanol (β-ZAL) and zearalanone, in blood serum; and (b) to conduct tests in species which are susceptible to ZEA to evaluate exposure to mycotoxicosis and to use serological biomarkers to assess the efficacy of AMA. In the first study, 24 beef heifers were randomly assigned to receive one of these treatments (n=6/treatament): T1) basal diet (control); T2) basal diet + 5 mg/kg ZEA; T3) basal diet + 5 mg/kg ZEA + 2.5 kg/t AMA; and T4) basal diet + 5 mg/kg ZEA + 5.0 kg/t AMA. The trial lasted 37 days. Blood was collected on different days after the diet was given, and the samples were centrifuged to obtain the blood serum and then analysed by HPLC-MS/MS. Among the analyzed metabolliates, β-ZAL was detected above the limit of quantification both in the unconjugated (>0.60 μg/kg) and conjugated (>1.70 μg/kg) forms. The remaining metabolites presented concentrations under the limit of detection. In the efficacy evaluation of the AMA, there was no significant difference (P>0.05) between the treatments with and without AMA at the tested levels of inclusion. In the second study, 70 pre-pubertal gilts were distributed in seven feeding groups (n=10/treatment) receiving diets with two levels of three clay-based AMAs (0.25% and 0.50%) and three levels of ZEA (0, 0.75 and 1 mg/kg) for 42 days. Initial and final body weight (BW), mean daily weight gain, mean daily feed intake, feed conversion and vulva volume were assessed at the time of blood sampling to determine ZEA and its metabolites in the serum. Blood samples were spun to obtain the serum, which was analyzed via HPLC-MS/MS. When the experimental period ceased, the animals were slaughtered and eviscerated in order to evaluate weight and length of the reproductive tract. The zootechnical performance was not affected by the presence of ZEA in the diet, and inclusion of AMA did not determine differences (P>0.05). ZEA caused a significant increase (P<0.05) in BW, length of the reproductive tract and vulva measurement (width, length and area), with no difference (P>0.05) between treatments with the addition of AMA to the diet (P>0.05). With respect to the evaluated metabolites, α-ZOL was detected above the limit of quantification in both unconjugated (>0.15 μg/kg) and unconjugated (>1.58 μg/kg) forms. The concentration of the remaining metabolites was below the limit of detection, thus confirming the results of the efficacy of AMA (P<0.05). The findings in this study contribute to the search of quality of the marketed AMA as well as to the assessment of analytical methods and biomarkers; these substances may represent an alternative to confirm the efficacy of such products in heifers and gilts. |
publishDate |
2020 |
dc.date.issued.fl_str_mv |
2020-02-14 |
dc.date.accessioned.fl_str_mv |
2021-09-15T10:49:56Z |
dc.date.available.fl_str_mv |
2021-09-15T10:49:56Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/doctoralThesis |
format |
doctoralThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://repositorio.ufsm.br/handle/1/22220 |
url |
http://repositorio.ufsm.br/handle/1/22220 |
dc.language.iso.fl_str_mv |
por |
language |
por |
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500500000007 |
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dc.rights.driver.fl_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ |
eu_rights_str_mv |
openAccess |
dc.publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Centro de Ciências Rurais |
dc.publisher.program.fl_str_mv |
Programa de Pós-Graduação em Medicina Veterinária |
dc.publisher.initials.fl_str_mv |
UFSM |
dc.publisher.country.fl_str_mv |
Brasil |
dc.publisher.department.fl_str_mv |
Medicina Veterinária |
publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Centro de Ciências Rurais |
dc.source.none.fl_str_mv |
reponame:Manancial - Repositório Digital da UFSM instname:Universidade Federal de Santa Maria (UFSM) instacron:UFSM |
instname_str |
Universidade Federal de Santa Maria (UFSM) |
instacron_str |
UFSM |
institution |
UFSM |
reponame_str |
Manancial - Repositório Digital da UFSM |
collection |
Manancial - Repositório Digital da UFSM |
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repository.mail.fl_str_mv |
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