BC-1606 Microbiologia

Aula 12 Diversidade Procariontes I

Prof . Antônio Sérgio Kimus Braz Santo André - Março de 2012

Importância da microbiologia na ...

Importância da microbiologia na ...

Dando uma mãozinha pra aves digerrem grãos Ou melhor dados umas enzimas Tolerantes a acidez , etc..

A biologia molecular se aproveita muito dos plasmídeos

Como isolar e identificar organismos ? Tarefa bastante complexa … Na maioria das vezes falhamos em cultivar organismos observados em um ambiente...

E para organismos especificamente organismos Patogênicos a situação é ainda mais complicada ...

Postulados de KOCH 1º O organismo patogênico deve estar presente Em todos casos da doença e ausente em animais sadios 2º O org patogênico deve ser cultivado em cultura pura. 3º células de uma cultura puro do organismo suspeito deve provocar a doença em animal sadio 4º o organismo deve ser isolado e caracterizado como mesmo encontrado originalmente

Abordagem atual .... Metagenômica...

Fluorescence in situ hybridization (CARD-FISH) of Eel River Basin sediment

Pernthaler A et al. PNAS 2008;105:7052-7057

©2008 by National Academy of Sciences

CARD-FISH of Eel River Basin sediment with probe ANME-2c_760 (green) and a general DNA stain (DAPI, blue), before (a) and after (b) capture with paramagnetic beads (dark red, collage of six images)

Pernthaler A et al. PNAS 2008;105:7052-7057

©2008 by National Academy of Sciences

Exemplo: Metagenômica de fluido vaginal Uso de sondas de RNA ribossomal 16s rRNA fluorescentes Com cores diferentes para cadas especie de bactéria Fluorescence image of vaginal fluid from a subject with BV. Bacteria are shown hybridizing with probes targeting BVAB1 (green) and Mobiluncus (red) and visualized by fluorescence in situ hybridization (FISH). Other bacteria (blue) are seen with 4′,6-diamidine-2-phenylindole, dihydrochloride, (DAPI), which stains DNA. The inset shows that Mobiluncus (green) is larger than BVAB1 (red) but has the same curved morphology. (With permission from D. N. Fredricks, T. L. Fiedler, and J. M. Marrazzo, “Molecular identification of bacteria associated with bacterial vaginosis,” New England Journal of Medicine, vol. 353, pp. 1899–1911, 2005.)

Quando surgiram ?

Arvore da vida … Diversidade dos seres vivos 3 Domínios ..

The tree of life stems from the last universal common ancestor (LUCA), which evolved into bacteria, archaea (microorganisms similar to bacteria) and eukaryotes (organisms with nucleus-containing cells). Extant bacteria are mostly mesophilic (they thrive at temperatures of about 25–40 °C), but some are thermophilic (preferring temperatures greater than about 60 °C). Many extant archaea are thermophilic, but no such eukaryotes exist. Gaucher et al.1 modelled and reconstructed proteins from ancestral bacteria, and from their heat stability conclude that the earliest bacteria lived in oceans at temperatures of 65–73 °C. Previous studies4, 5 based on models of ancient RNA suggest that LUCA was either mesophilic or thermophilic. The temperature at which the earliest archaea flourished has not been estimated.

Bacterias

Archaea

eucariontes

Bacterias

3 filos Hipertermofilas De ramificação precoce

Hipertermofilas de ramificação precoce

Thermotoga Bacilo hipertermofilo (cresce a 90ºC) Envoltorio tipo bainha => toga Anaeróbico Quimiorganotrofico fermentativo : Quebra açucares e polimeros Produzindo acetato, CO2 e H2 Ou respira anerobicamente H2 doando eletrons para Fe3+ Habitat: Isolados de fontes termais terrestres e fendas hidrotermais 20 % do seu genoma deve ter vindo de Archaea hipertermofilas

Hipertermofilas de ramificação precoce

Thermodesulfobacterium: Termofilica => Thermo Redutora de sulfato => desulfo bacterium

Temperatura ideal 70ºC Anaerobico estrito Quimiorganotrofico (respiração anaeróbica S)

Usa lactato, piravato, etanol Para reduzir SO4²⁻ a H2S Habitat: Isolados de fontes termais terrestres e fendas hidrotermais

Thermodesulfobacterium

Aspectos de adaptação de lipídeos similares a arqueas Mas não identico – ao fitanil

Aquifex , Thermocrinus (e relacionados) hipertermofilo (cresce até 95ºC – ideal 85ºC) Microareofilo Alguns como Hidrogenobacter são aerobicos obrigatorios Autotrofico Quimilitotrofico Usam H2 S⁰ e S2O3²⁻ como doadores de elétrons (oxidam) *Não crescem quimiorganotroficamente nem em ambientes ricos Genoma muito pequeno 1/3 da E coli 16% de genes oriundos de Archaea (transferencia horizontal) Habitat: Isolados de fontes termais terrestres e fendas hidrotermais

2 H2 + O2 → 2 H2O

83ºC pH 8

Thermocrinus Primerio termófilo a ser descoberto década de 60 Por Brock ( autor original do livro texto do curso...)

Bacterias verdes não sulfurosas Chloroflexus e Heliotrix Fototrófico anoxigênico Termofílico (metabolismo flexível!!!) Fotoautotrofico H2S +CO2 H2 +CO2 Mas cresce melhor como fotoheterotrofico (Fonte organica de carbono) E cresce bem também como quimiorganotrófico e fazendo respiração aeróbica

Bacterias verdes não sulforosas Thermomicrobium Quimiotrofico Oxida compostos organiscos e tb CO (monoxido de carbono) Estritamente aeróbico Termófilo (75ºC ideal) > parede celular sem peptideoglicano (somente proteínas) > membrana com Lipidio atípico (alcool)

Lipidido incomuns de Thermomicrobium

Adaptação a temperaturas elevadas

Flagellum-related genes all located on the megaplasmid

Deinococos Deinococcus

Quimiorganotrofico Necessita meio complexo para crescimento Encontrado em solo, fezes, carne, poeira, extrato de ostra , etc.. Resistência estrema a radiação

Ambiente natural desconhecido 2 cromossomos ! e 1 mega plasmídeo e vários pequenos plasmídeos

* Parede celular atípica , sem DAP , em vez disso tem ornitina no lugar

Parede celular complexa Coloração gram + So que estrutura semelhante a gram Varias camadas de parede celular *Não tem lipideo A na membrana externa

Muitos carotenoides Coloração alaranjada

Mais resistente a radiação que endosporos Resistem a 1500 x mais radiação do que um humano Altamente eficiente no reparo Mesmo contra agentes mutagénicos químicos

Published by AAAS

J. Lin et al., Science 285, 1558-1562 (1999)

Figure 3. DNA repair after irradiation, visualized by optical mapping. DNA samples were extracted at different time points from D. radiodurans R1 (wild type) cells. The irradiation and recovery conditions (time interval after irradiation) for each image were: (A) 0 Gy, 0 hours; (B) 17.5 Gy, 0 hours; (C) 17.5 Gy, 2 hours; (D) 17.5 Gy, 4 hours; (E) 17.5 Gy, 6 hours; (F) 17.5 Gy, 16 hours; (G) 17.5 Gy, 24 hours. Scale bar, 20 µm. Histogram analysis (34) showed consistent elongation of DNA molecules into full-length genomic molecules. The recA-deficient strain (rec 30) showed no evidence of repair, and the observable number of molecules decreased with time as a result of cell death (35)

Deinococcus geothermalis quimiotrofico Reduz Fe (III)-nitrilotriacetic acid, U (VI), Cr (VI). Hg(II) Resite a radiação e temperaturas elevadas 1 cromossomo apenas

Formam biofilmes Em metais Problemas Em industrias de Papel Deinococcus geothermali Meiothermus silvanus

Thermus Termofilo até 79 ºC (ideal 75ºC) Quimiorganotrofico Aerobico (incapaz de fazer fermentação)

Thermus aquaticus (Taq DNA polimerase)

Espiroquetas Gram Intensamente espiraladas (CreS) Flexíveis e delgadas, Muito moveis Amplamente disseminadas em > Ambientes aquáticos Vida livre > dentro de animais Algum são patogenicos outros simbiontes Podem ser aerobicos , microarefilos e anerobicos

Caracteristica propria endoflagelo

Borrelia Brachyspira Brevinema Clevelandina Cristispira Diplocalyx Hollandina Leptonema Leptospira Pillotina Serpulina Spirochaeta Treponema

Treponema

Flagelos em espiroquetas

(espiroqueta)

Salmonella

(flagelo normal)

Flagelo periplasmático

Nature, Murphy et al, 2006.

Flagelo

Membrana externa Cilindro citoplasmático

Flagelo periplasmático Membrana externa Flagelo

Espaço periplasmático

Peptídeoglicana Membrana citoplasmática

Proteínas embebidas na membrana externa

Flagelo periplasmático FlaB

Flagelo ancorado na membrana celular

FlaA

Spirochaeta e Cristispira Spirochaeta : vida livre, ambientge aquaticos, lodo, rios, lagos, lagoas oceanos.. Anaerobios e aerobicos podem fazer fermentação Um tipo incomum é simbionte de cupins O metabolismo de celulose produz H2 e CO2 A spirochaeta usa H2 e CO2 e converte em acetato Podem fixar N2 atmosferico Cristispira ajuda digestão de moluscos, fica estilete cristalino destes

Os “flagelos” desse Eucarionte Simbionte de cupin ( e que digere celulose) São na verdade spiroquetas

Karyomastigont model of origin of nucleated cells. The LECA evolved from eubacterial-archaebacterial syntrophies in which sulfideoxidizing spirochetes attached to sulfidogenic thermoplasmas to form a “Thiodendron”-like consortium. Archaeprotist (trichomonad Mixotricha paradoxa, lower right), LECA analogue in termite Mastotermes darwiniensis, swims via motility symbiosis with 200,000 Treponema sp. surface spirochetes. Four distinctive surface spirochetes are detected by morphological and molecular techniques (36). Chromatin appears first in the karyomastigont (“kymstgnt”), the precursor cytoskeletal organellar system from which the tethered nucleus (“n”) was released.

Borrelia burgdorferi => doença do lyme

Possui um cromosso pequeno linear

Vetor carrapatos

Leptospira leptospirose

amarelo

Treponema pallidum

Sífilis A sífilis também é conhecida como lues (palavra latina que significa praga), cancro duro, avariose, doença-do-mundo, mal-de-franga, mal-de-nápoles, lues, mal-de-santa-eufêmia e pudendagra, entre outros..

é uma doença infecciosa evolui lentamente em três estágios, e caracterizada por lesões da pele e mucosas. Pode ser transmitida por contato sexual, configurando-se assim como uma DST, e mais raramente por contaminação feto-placentária. O Treponema pallidum é uma bactéria com forma de espiral (em média dá 10 a 126 voltas) e tem cerca de 54 micrómetros de comprimento mas apenas 0,2 micrómetros

Primary Stage The primary stage of syphilis is usually marked by the appearance of a single sore (called a chancre), but there may be multiple sores. The time between infection with syphilis and the start of the first symptom can range from 10 to 90 days (average 21 days). The chancre is usually firm, round, small, and painless. It appears at the spot where syphilis entered the body. The chancre lasts 3 to 6 weeks, and it heals without treatment. However, if adequate treatment is not administered, the infection progresses to the secondary stage. Secondary Stage

Skin rash and mucous membrane lesions characterize the secondary stage. This stage typically starts with the development of a rash on one or more areas of the body. The rash usually does not cause itching. Rashes associated with secondary syphilis can appear as the chancre is healing or several weeks after the chancre has healed. The characteristic rash of secondary syphilis may appear as rough, red, or reddish brown spots both on the palms of the hands and the bottoms of the feet. However, rashes with a different appearance may occur on other parts of the body, sometimes resembling rashes caused by other diseases. Sometimes rashes associated with secondary syphilis are so faint that they are not noticed. In addition to rashes, symptoms of secondary syphilis may include fever, swollen lymph glands, sore throat, patchy hair loss, headaches, weight loss, muscle aches, and fatigue. The signs and symptoms of secondary syphilis will resolve with or without treatment, but without treatment, the infection will progress to the latent and possibly late stages of disease. Latent and Late Stages The latent (hidden) stage of syphilis begins when primary and secondary symptoms disappear. Without treatment, the infected person will continue to have syphilis even though there are no signs or symptoms; infection remains in the body. This latent stage can last for years. The late stages of syphilis can develop in about 15% of people who have not been treated for syphilis, and can appear 10-20 years after infection was first acquired. In the late stages of syphilis, the disease may damage the internal organs, including the brain, nerves, eyes, heart, blood vessels, liver, bones, and joints. Signs and symptoms of the late stage of syphilis include difficulty coordinating muscle movements, paralysis, numbness, gradual blindness, and dementia. This damage may be serious enough to cause death.

Portrait of Gerard de Lairesse by Rembrandt van Rijn, ca. 1665–67, oil on canvas. De Lairesse, himself a painter and art theorist, suffered from congenital syphilis that severely deformed his face and eventually blinded him

Não existia na europa até a descoberta da Américas.. Essa doeça foi levada para europa pro colombo … (por outro lado os europeus levaram um numero bem maior de patogenos para o novo mundo...)

Tuskegee and Guatemala studies See also: Tuskegee syphilis experiment and Syphilis experiments in Guatemala One of the most infamous United States cases of questionable medical ethics in the 20th century was the Tuskegee syphilis study.[44] The study took place in Tuskegee, Alabama, and was supported by the U.S. Public Health Service (PHS) in partnership with the Tuskegee Institute.[45] The study began in 1932, when syphilis was a widespread problem and there was no safe and effective treatment.[6] The study was designed to measure the progression of untreated syphilis. By 1947, penicillin had been validated as an effective cure for syphilis and was becoming widely used to treat the disease. Study directors, however, continued the study and did not offer the participants treatment with penicillin.[45] This is debated, and some have found that penicillin was given to many of the subjects.[6] The study did not end until 1972.[45] Syphilis experiments were also carried out in Guatemala from 1946 to 1948. They were United Statessponsored human experiments, conducted during the government of Juan José Arévalo with the cooperation of some Guatemalan health ministries and officials. Doctors infected soldiers, prisoners, and mental patients with syphilis and other sexually transmitted diseases, without the informed consent of the subjects, and then treated them with antibiotics. In October 2010, the U.S. formally apologized to Guatemala for conducting these experiments.[46]

Treponema pertenue => Bouba (também conhecida como frambesia, parangi, patek, piã) é uma doença tropical infecciosa da pele, ossos e cartilagens causada pela bactéria espiroqueta