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COLETÂNEA DE PROCEDIMENTOS TÉCNICOS E METODOLOGIAS EMPREGADAS PARA O ESTUDO DE Bacillus E GÊNEROS ESPORULADOS AERÓBIOS CORRELATOS

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COLETÂNEA DE PROCEDIMENTOS TÉCNICOS E METODOLOGIAS EMPREGADAS PARA O ESTUDO DE Bacillus E GÊNEROS ESPORULADOS AERÓBIOS CORRELATOS Instituto Oswaldo Cruz Laboratório de Fisiologia Bacteriana COLETÂNEA DE PROCEDIMENTOS TÉCNICOS E METODOLOGIAS EMPREGADAS PARA O ESTUDO DE Bacillus E GÊNEROS ESPORULADOS AERÓBIOS CORRELATOS AUTORES Leon Rabinovitch Edmar Justo de Oliveira 2015 Laboratório de Fisiologia Bacteriana COLETÂNEA DE PROCEDIMENTOS TÉCNICOS E METODOLOGIAS EMPREGADAS PARA O ESTUDO DE Bacillus E GÊNEROS ESPORULADOS AERÓBIOS CORRELATOS Laboratório de Fisiologia Bacteriana – LFB (Coleção de Culturas do Gênero Bacillus e Gêneros Correlatos – CCGB e Laboratório de Referência Nacional para Carbúnculo – LARENAC) Laboratório de Fisiologia Bacteriana - LFB Coleção de Culturas do Gênero Bacillus e Gêneros Correlatos - CCGB Laboratório de Referência Nacional para Carbúnculo - LARENAC Tels: 2562-1640/2562-1637/2562-1639 Pavilhão Rocha Lima, 3º andar, salas 300-308-310-312 Av. Brasil, 4365, Manguinhos Rio de Janeiro – RJ CEP: 21040-900 e-mail: [email protected] Dados Internacionais de Catalogação-na-Publicação (CIP) Col683 Coletânea de procedimentos técnicos e metodologias empregadas para o estudo de Bacillus e gêneros esporulados aeróbios correlatos / Autores: Leon Rabinovitch e Edmar Justo de Oliveira. – 1. Ed. – Rio de Janeiro : Montenegro Comunicação, 2015. 160p. ; 21x28cm. Inclui bibliografia. ISBN 978-85-67506-04-3 (broch.) 1. Bacteriologia. I. Rabinovitch, Leon. II. Oliveira, Edmar Justo. CDD 610.8 Laboratório de Fisiologia Bacteriana COLETÂNEA DE PROCEDIMENTOS TÉCNICOS E METODOLOGIAS EMPREGADAS PARA O ESTUDO DE Bacillus E GÊNEROS ESPORULADOS AERÓBIOS CORRELATOS O Laboratório de Fisiologia Bacteriana, LFB, do Instituto Oswaldo Cruz, da Fundação Oswaldo Cruz é integrado por seto- res de pesquisa, ensino e desenvolvimento tecnológico, tendo na sua estrutura o Laboratório de Referência Nacional para Carbúnculo, LARENAC e Coleção de Culturas do Gênero Bacillus e Gêneros Correlatos, CCGB, filiada à Federação Mundial de Coleções de Culturas, WFCC. AUTOR-ORGANIZADOR: Leon Rabinovitch, Farmacêutico-Químico, PhD AUTOR ASSOCIADO Edmar Justo de Oliveira, Químico, MSc COLABORADORES: Adriana Marcos Vivoni, Microbiologista, PhD Claude André Solari, Médico, PhD Claudio Simões de Mattos, Biomédico, MSc Jeane Quintanilha Chaves, Farmacêutica, PhD Josiane Teixeira de Brito, Bióloga Sônia Ermelinda Alves da Silva, Bióloga, MSc Vera Cristina Pessoa de Lima, Bióloga Primeira Edição - 2015 Imagens de capa: Bacillus, célula vegetativa e esporângios com corpo paraesporal. SUMÁRIO COLETÂNEA DE PROCEDIMENTOS TÉCNICOS E METODOLOGIAS EMPREGADAS PARA O ESTUDO DE Bacillus E GÊNEROS ESPORULADOS AERÓBIOS CORRELATOS PREFÁCIO .................................................................................................................................................... 11 INTRODUÇÃO ............................................................................................................................................. 12 NOÇÕES E CONCEITOS BÁSICOS ........................................................................................................... 13 CONCEITOS DE ESPÉCIE EM CÉLULAS PROCARIÓTICAS COMO AS BACTÉRIAS ...................... 14 BACTÉRIAS COMO SISTEMAS VIVOS ................................................................................................... 15 BASTONETES E COCOS PRODUTORES DE ENDÓSPOROS ................................................................ 16 TABELA 1: ALGUNS CARACTERES DE BACTÉRIAS FORMADORAS DE ENDÓSPOROS E GÊNEROS CORRELATOS ............................................................................................ 18 MORFOLOGIA CELULAR .......................................................................................................................... 19 PAREDE CELULAR DE Bacillus E GÊNEROS CORRELATOS ............................................................... 21 TABELA 2: ALGUNS TIPOS DE MUREÍNA ENCONTRADOS EM Bacillus E EX-Bacillus HOJE PERTENCENTES A OUTROS GÊNEROS ............................................................... 22 CÁPSULAS ................................................................................................................................................... 24 DIFERENCIAÇÃO CELULAR EM Bacillus E GÊNEROS CORRELACIONADOS1 ............................... 25 TABELA 3: CITOMORFOLOGIA DE Bacillus E GÊNEROS CORRELATOS. POSIÇÃO DE ESPOROS NO ESPORÂNGIO E DIMENSÕES ................................................................. 26 TABELA 3A: CITOMORFOLOGIA DE Bacillus E GÊNEROS CORRELATOS. POSIÇÃO DE ESPOROS NO ESPORÂNGIO E DIMENSÕES ................................................................. 27 TABELA 3B: CITOMORFOLOGIA DE Bacillus E GÊNEROS CORRELATOS. POSIÇÃO DE ESPOROS NO ESPORÂNGIO E DIMENSÕES ................................................................. 28 TABELA 4: EXEMPLO DE ALGUMAS CARACTERÍSTICAS DIFERENCIAIS PARA A SEPARAÇÃO DOS GÊNEROS Bacillus, Geobacillus, Paenibacillus e Virgibacillus ................. 30 PROCEDIMENTOS MÍNIMOS PADRONIZADOS PARA A DESCRIÇÃO DE NOVA ESPÉCIE DO GÊNERO Bacillus E BACTÉRIAS RELACIONADAS ..................................... 32 1. EXIGÊNCIA GERAL - PROCEDIMENTOS ..................................................................................... 32 2. CARACTERES MORFOLÓGICOS E TINTORIAIS ........................................................................ 33 3. CARACTERES COLONIAIS ............................................................................................................. 33 Laboratório de Fisiologia Bacteriana 5 4. CARACTERISTICAS FISIOLÓGICAS ............................................................................................ 33 5. CARACTERÍSTICAS BIOQUÍMICAS AVALIADAS ATRAVÉS DAS SEGUINTES PROVAS ... 34 6. CARACTERES QUIMIOTAXONÔMICAS ...................................................................................... 35 7. ESTUDOS DOS ÁCIDOS NUCLÉICOS ........................................................................................... 35 CAPÍTULO 1: MEIOS PARA CULTIVO E ISOLAMENTO ..................................................................... 36 1.1. CALDO NUTRIENTE – CN ........................................................................................................... 36 1.2. ÁGAR NUTRIENTE – AN .............................................................................................................. 36 1.3. ÁGAR NUTRIENTE – ANCTC ...................................................................................................... 36 1.4. CALDO-J ......................................................................................................................................... 37 1.5. ÁGAR-J ........................................................................................................................................... 37 1.6. ÁGAR-J DIFÁSICO ........................................................................................................................ 37 1.7. ÁGAR EXTRATO DE SOLO – AES .............................................................................................. 38 1.8 EXTRATO DE SOLO ....................................................................................................................... 38 1.9. MEIO SABOURAUD DEXTROSE. VERIFICAÇÃO DO CRESCIMETO EM PH 5,7 ............... 39 1.10. MEIO CALDO GLICOSADO – VP E PARA O TESTE DE VOGES-PROSKAUER ................. 40 1.11. MEIO NYSM ................................................................................................................................. 42 1.12. MEIO NYSM – SÓLIDO ............................................................................................................... 42 1.13. MEIO DE CIANETO DE POTÁSSIO .......................................................................................... 42 1.14. MEIO DE HIPURATO .................................................................................................................. 43 1.15. VERIFICAÇÃO DO CRESCIMENTO EM PRESENÇA DE CALDO AZIDA-SÓDICA .......... 44 1.16. MEIO VRM PARA CONTAGEM DE Bacillus cereus ................................................................. 45 1.17. CALDO MBS ................................................................................................................................ 46 1.18. ÁGAR MBS ................................................................................................................................... 46 1.19. MEIO ANM LÍQUIDO .................................................................................................................. 47 1.20. MEIO PARA Bacillus cereus BCM ............................................................................................... 47 1.21. ÁGAR ANAERÓBICO ................................................................................................................. 48 1.22. ÁGAR MUELLER HINTON ........................................................................................................ 49 1.23. ÁGAR PLATE COUNT (PCA) .................................................................................................... 49 CAPÍTULO 2: PROVAS BIOQUÍMICAS .................................................................................................. 50 2.1. MEIO DE NITRATO ....................................................................................................................... 50 6 Laboratório de Fisiologia Bacteriana 2.2. MEIO DE FENILALANINA ........................................................................................................... 50 2.3. MEIO DE GELATINA
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