MICROBIOLOGA

Blgo, Xavier lvarez Montero, Ph.D.

UNIDAD # 1
Teora Prctica
Generalidades Medios de cultivo
Dominio Bacteria Aislamiento de bacterias
Dominio Archaea Medicin del crecimiento
UNIDAD # 2
Dominio Eucaria Medios de cultivo
Anlisis de metabolitos
Fermentacin fngica
UNIDAD # 3
Microorganismos patgenos
Introduccin
Generalidades
MICROBIOTA HUMANA APARATO DIGESTIVO 1,3 kg
cerebro

Clulas microbianas 10 veces ms clulas humanas

MICROBIOMA codifica 8M genes

GENOMA codifica 20-25K genes

Escherichia coli
 Metabolizan los componentes de la dieta

Sintetizan nutrientes y vitaminas esenciales

Excretan metabolitos efectos antiinflamatorios y/o antioxidantes

Firmicutes, Bacteroidetes, Actinobacteria y Proteobacteria, Archea,

Virus y Hongos
Composicin del microbioma:
Propensin a la obesidad
Diabetes
Cncer colorrectal,
Colitis ulcerosa;
Esquizofrenia
Depresin,
 Ansiedad,

Autismo,
Prkinson,
Sndrome de Intestino Irritable (SII)

Enfermedades autoinmunes
Alergias

Enfermedad de Crohn
 Mediante la produccin de hormonas o molculas que generan al
alimentarse, modifican la qumica del cerebro
Interacta con el sistema inmunolgico
Diversidad es la clave: caf y yogur positivos; mayonesa, helados,
aceite de palma, snacks, ansiolticos, anticidos, antidepresivos, antibiticos
o comer demasiado negativos
Se considera Microbioma rgano
 El Trasplante Fecal de Microbiota (FMT) transferencia de materia fecal de un
individuo sano a uno enfermo para restablecer el equilibrio alterado a causa de
alguna enfermedad, infeccin o compuesto qumico. La nueva microbiota
procedentes de FMT es capaz de recolonizar el intestino del paciente, restablecer
el equilibrio del ecosistema intestinal

G.E. Hong siglo IV; E.E.U.U Clostridium difficile 96%

Patgenos prioritarios para la I+D de nuevos
antibioticos
Prioridad crtica:

1. Acinetobacter baumannii resistentes a los carbapenmicos (RC).

2. Pseudomonas aeruginosa RC.

3. Enterobacteriaceae RC; productoras de betalactamasas de espectro

extendido (ESBL).
 Prioridad alta:

1. Enterococcus faecium resistente a la vancomicina (RV).

2. Staphylococcus aureus resistente a la meticilina; RV.

3. Helicobacter pylori resistente a la claritromicina.

4. Campylobacter spp. resistente a las fluoroquinolonas (RQ).

5. Salmonella RQ.

6. Neisseria gonorrhoeae resistente a las cefalosporinas; RQ.

 Prioridad media:
1. Streptococcus pneumoniae sin sensibilidad a la penicilina.

2. Haemophilus influenzae resistente a la ampicilina.

3. Shigella spp. RQ.
4. 2050 10 millones + cncer.
 Vibrio cholera agua y condiciones higinicas deficientes periodo de
incubacin 2-120 h 47 pases 2,9M contagiados 95K muertes
2000M US$ ao-1 vacunas

Mycobacterium tuberculosis & M. bovis aire, animales estado latente 1 de

cada 3 personas (portadores) 10% desarrolla la enfermedad 1,8M
muertes OMS 2030 95% muertes (2182) 16000M ao-1

Multidrug and Extensively Drug-Resistant Tuberculosis (MEDRT)

Pandemias VIH/SIDA; Malaria

 >1% Bacterias
patgenas

<99% Bacterias
no patgenas

Medicina Medio ambiente Agricultura

Metabolitos Biorremediacin Ciclo y absorcin

bioactivos Bioenerga de nutrientes
 Virus
?

Resumen del desarrollo temporal de la vida en la

Tierra (Last Universal Common Ancestor)
 Microorganismos

Procariotas Eucariotas

Microalgas
Bacteria Archea Hongos
Protozoarios
 $)"15&3

t
MICROORGANISMS AND MICROBIOLOGY 7

DNA

Gene encoding 3. Sequence DNA. Aligned rRNA

UNIT 1
ribosomal RNA gene sequences
3

A G T CGC T A G 1
Cells A T T C CG T A G 2 1
1. Isolate DNA 2. Make copies 4. Analyze A GC CG T T A G 3 5. Generate
from each of rRNA gene sequence. phylogenetic
organism. by PCR. tree. 2
A G C T A A G
(a)

BACTERIA ARCHAEA EUKARYA

Macroorganisms
Animals
Entamoebae Slime
Green nonsulfur molds
bacteria Euryarchaeota
Fungi
Methanosarcina
Mitochondrion
Methano- Extreme Plants
Gram- Crenarchaeota bacterium
halophiles
Proteobacteria positive Thermoproteus
Ciliates
bacteria
Chloroplast Pyrodictium Thermoplasma
Cyanobacteria Thermococcus
Flagellates
Nitrosopumilus Pyrolobus
Green sulfur Methanopyrus
bacteria Trichomonads

Thermotoga

Thermodesulfobacterium Microsporidia

Aquifex Diplomonads

(b)

Figure 1.6 Evolutionary relationships and the

rbol filogentico
phylogenetic tree of life.(a) de los behind
The technology seres vivos deducido algorithm makes de pairwisela secuenciacin
the sequence aligned with sequences from other
organisms. A computer comparativa
versus organism 2, three differences; 1 versus 3,
two differences; 2 versus 3, four differences. Thus del
ribosomal RNA gene phylogenies. 1. DNA is extracted comparisons at each base and generates a phylogenetic organisms 1 and 3 are closer relatives than are 2 and
gen del ADN ribosomal
from cells. 2. Copies of the gene encoding rRNA are
made by the polymerase chain reaction (PCR;
tree, 5, that depicts evolutionary relationships. In the
example shown, the sequence differences are
3 or 1 and 2. (b) The phylogenetic tree of life. The tree
shows the three domains of organisms and a few
Dominio bacteria
ADN mlecula larga de dos cadenas (cromosoma) nucleoide cerrado
covalentemente y es circular

Una sola copia de cada gen haploides

ADN extra-cromosmico circular plsmidos propiedades especiales

(metablicas)

Escherichia coli 4,68Mpb 4300 genes 1900 tipos de protenas 2,4M

protenas totales
in microbiology: transmission and scanning.
Transmission Electron Microscopy
The transmission electron microscope (TEM) is used to examine
cells and cell structure at very high magnification and resolu-
tion. The resolving power of a TEM is much greater than that
which are then examined individually by TEM (Figure 2.10a)
To obtain sufficient contrast, the sections are treated with stains
such as osmic acid, or permanganate, uranium, lanthanum, or
lead salts. Because these substances are composed of atoms of
high atomic weight, they scatter electrons well and thus improve
contrast. If only the external features of an organism are to be

Cytoplasmic Septum Cell wall DNA

membrane (nucleoid)

Stanley C. Holt
(a)

Microfotografa de una clula bacteriana en divisin tomada con un

microscopio electrnico de transmisin. La clula mide 0,8 m de ancho
Bacterias fototrficas

Conservar energa lumnica Tierra anxica

Desarrollo de la fotosntesis bacterias fottrofos anoxignicos no O2

H2O donador de electrones H2; H2S; Fe+2

Proteobacteria; Chlorobi; Chloroflexi; Firmicutes; Acidobacteria Fotosntesis

Anoxignica (FA)

Fotosntesis oxignica (FO) cianobacterias

 Bacterias fototrficas pigmentos similares a la clorofila + pigmentos accesorios
para recolectar energa lumnica centro de reaccin membrana
transferencia de e- ATP

Bacterias fototrficas anoxignicas Fotosistema I (FI) o Fotosistema II (FII)

Fotosistema I Fotosistema I (FO); Fotosistema II Fotosistema II (FO)

Fotosistema I & II cianobacterias

Cianobacterias

Unicelulares & filamentosas

0,5-100 m dimetro

Hormogonias

Acinetos

Heterocistos

Gneros: Synechococcus, Cyanothece, Lyngbya, Arthrospira, Oscillatoria, Nostoc

 Reproduce by formation eter
tous to
of cells(Figure
forms as large14.2c);
Dermocarpa, as Xenococcus,
100(4)m in diameter.
Nostocales Cyanobacteria
are filamentous, can
divide fission in a single plane
Phylogeny
The and Classification
morphological diversity of theof Cyanobacteria
Cyanobacteriais impressive baeocytes produced
small spherical through
cells called
be divided
Chroococcidiopsis
Dermocarpella,
into five Myxosarcina,
morphological groups: (1) Chroococcales are
Group (The morphological
Figure Genera
diversity
14.2). Both unicellular of
andthe Cyanobacteria
filamentous formsisareimpressive
known,
multiple
baeocytes fission along a singleChroococcidiopsis
produced through
unicellular,
axis, and are capable of cellular differentiation (Fig-
dividing by binary fission (Figure 14.2a); (2) Pleuro- Group IV,
Group V, Stigonematales.
Nostocales. Cells
and
multiple
Group fission
III, Oscillatoriales. ure 14.2d);Lyngbyaand (5) (FigureStigonematales
14.2c), Spirulinaare morphologically similar divide to formcells
branches
(Figure 14.2).
there is Both unicellular
considerable and filamentous
variation within these forms are known,
morphological capsales are(Figure
unicellular, dividing by inmultiple Filamentous that produce
Group I, Chroococcales.
types. Cyanobacterial cellsGloeothece
range in within
size from (Figurem in14.2a),
0.5morphological
diam-
Undifferentiated
Group III, Oscillatoriales. to
filamentous Nostocales except
Lyngbya 14.5),
(Figure that cellsSpirulina
Arthrospira,
14.2c), divide multiplefission
planes(colonial)
forming heterocysts
and there is considerable variation these cells that divide by (Figure
binary 14.2b); (3)
OscillatoriaOscillatoriales
(Figure 14.6ab),are filamentous nonheterocys-
eter to Cyanobacterial
cells as large ascells
100 range
m inindiameter. Cyanobacteria can
Undifferentiated filamentous branching filaments
(Figure 14.5), (Figure 14.2e).
Arthrospira, Finally, the prochlorophytes
types. size from 0.5 m in diam-
Unicellular or cell aggregates
be
eterdivided
to cellsinto five morphological
as large
Gloeobacter,
as 100 m in groups:
diameter.
Synechococcus,
(1)Cyanobacteria
Chroococcales can are
fission in adivide
cells that singlebyplane tous
are a forms
binary lineage(Figure
fission in a single planealong a single
Microcoleus,
Oscillatoria
of unique
Trichodesmium
14.2c); (4) Nostocales
Pseudanabaena,
(Figure 14.6ab),
unicellular
(Figure 14.7b)
are filamentous,
Cyanobacteria divide
once thought to
be distinct butaxis,
Microcoleus,
now and are
classified capable
Pseudanabaena,
within of cellular
the differentiation
Chroococcales. (Fig- not (Figure 14.3). Species of Pl
by binary Cyanothece, Gloeocapsa, Table 14.1 Group V, Stigonematales. Cells
unicellular,
be divided into dividing
five morphological fission (Figure
groups: (1)14.2a); (2) Pleuro-
Chroococcales are Trichodesmium (Figure), 14.7b)

Susan Barns and Norman Pace

Group IV, Nostocales. ure 14.2d); Nodularia
and (5) (Figures 14.2d
Stigonematales Nostoc,
are morphologically similar
lists some major genera currently recognized in each group. divide
within theto cyanobacteria,
form branches in
capsales
unicellular,aredividing
unicellular, dividing
by binary by multiple
fission fission(2)
(Figure 14.2a); (colonial)
Pleuro- Filamentous cells that produce Calothrix (Figure 14.8ab),
(Figure
capsales14.2b); (3) Oscillatoriales
are unicellular,
Synechocystis,
dividingare by filamentous
Chamaesiphon,
nonheterocys-
multiple fission (colonial)
Group IV, Nostocales.
heterocysts
to
Filamentous cells that produce
Nostocales
Some of the
branching
Nodularia
except
Anabaena
Calothrix
filaments
(Figures
major(Figure that
(Figure
(Figure
14.2d
cells
morphological
),
14.6c),
14.8ab),
Nostoc,
divide
14.2e).
in multiple planes forming
classifications of Cyanobac-
the prochlorophytes
Finally, groups,
tiple fission arose only once
teria correspondCylindrospermum,
to coherent Scytonema,
phylogenetic but others do nobacteria (Figure 14.3). Lik
Merismopedia, Crocosphaera

Daniel H. Buckley
heterocysts Anabaena (Figure 14.6c),
tous
(Figureforms (Figure
14.2b); 14.2c); (4) Nostocales
(3) Oscillatoriales are filamentous,
are filamentous divide
nonheterocys- are a lineageRichelia
of unique (Figure unicellular
14.7c) Cyanobacteria once thought to
Cylindrospermum, Scytonema,
along a single
tous forms axis, and
(Figure are capable
14.2c); of cellular
(4) Nostocales are differentiation
filamentous, divide(Fig- beCells
distinctFischerella
but now(Figures
classified within
14.8c,the
d ), Chroococcales. Table 14.1
ure
along 14.2d);
a singleand (5)and
axis, (Figure
Stigonematales
are capable ofare 14.7a),
morphologically
cellular Prochlorococcus,
differentiation similar
(Fig-
Group V, Stigonematales.
form brancheslists
divideV,toStigonematales.
Richelia
some Fischerella
major genera
Stigonema,
(Figure 14.7c)
14.2e,
currently
Chlorogloeopsis, recognized in each group.
not (Figure 14.3). Species of Pl
within the cyanobacteria, in
Group Cells (Figures 14.2e, 14.8c, d ),
to
ureNostocales
14.2d); and except Prochloron
that cells divide
(5) Stigonematales in multiple
are planes forming
morphologically similar divide to form branches (a) Some of the Hapalosiphon
major morphological
Stigonema, Chlorogloeopsis, classifications (b) of Cyanobac- tiple fission arose only
(c) once
branching
to Nostocales filaments
except (Figure 14.2e).
that cells divide Finally, the prochlorophytes
in multiple planes forming teria correspond Hapalosiphon
to coherent phylogenetic groups, but others do nobacteria (Figure 14.3). Lik
Group II, Pleurocapsales.
are a lineage
branching
be
of unique
filaments Pleurocapsa
unicellular
(Figure 14.2e).Cyanobacteria (Figure
Finally, the prochlorophytes
once thought 14.2b),
to
aredistinct
a lineagebutof now
uniqueclassified within
unicellular the Chroococcales.
Cyanobacteria once thought to
Table 14.1 not (Figure 14.3). Species of Pleurocapsales form a coherent group
Reproduce by formation
lists some of
major genera Dermocarpa,
currently recognized in each
be distinct but now classified within the Chroococcales. Table 14.1 Xenococcus,
group. within the 14.3
not (Figure cyanobacteria,Figure
). Species 14.2 Cyanobacteria:
of indicating
Pleurocapsales that form the
a five
reproduction majorbygroup
coherent mul-

PaceBarns and Norman Pace

Some of the major morphological classifications of Cyanobac- tiple fission arose morphological
only once in types
the of cyanobacteria.
evolutionary history of cya-
small spherical cells
teria
called
lists some major genera currently recognized in each group.
correspond to coherent
Dermocarpella,
phylogenetic groups,
Some of the major morphological classifications of Cyanobac- but
Myxosarcina,
others do
within the cyanobacteria, indicating that reproduction by mul-
nobacteria (Figure
tiple fission arose only
(a) Unicellular, Gloeothece; a single cell measures
14.3). Likewise,
once in the species
evolutionary of the Nostocales
history of cya- and
note to COMP:
baeocytes produced through Chroococcidiopsis 56 m in diameter; (b) colonial, Pleurocapsa; these

Daniel H. Buckley
teria correspond to coherent phylogenetic groups, but others do nobacteria (Figure 14.3). Likewise,
structures speciesofofcells
contain hundreds theandNostocales
are 7 50 and
Please put figure
multiple fission m in diameter; (c) filamentous, Lyngbya; a single

Susan

Daniel H. Buckley
cell measures
legend aboutin10 space
m wide; (d)
in filamentous

Susan Barns and Norman

heterocystous,
(a) Nodularia; a single cell measures about (b) (c)
Group III, Oscillatoriales. Lyngbya (Figure 14.2c), Spirulina 10 m wide;
lower(e) filamentous branching, Fischerella;
left corner

Daniel H. Buckley
Undifferentiated filamentous (Figure 14.5), Arthrospira, a cell is about 10 m wide. See how morphological
diversity relates to phylogenetic diversity in Figure 14.3. (d) (e)
cells that divide by binary Oscillatoria (Figure 14.6ab),
Pace Pace
Figure 14.2 Cyanobacteria: the five major

and Norman
morphological types of cyanobacteria.
(a) (b) (c)
fission in a single plane Microcoleus, Pseudanabaena, and Norman (a) Unicellular, Gloeothece; a single cell measures

H. Buckley

H. Buckley
note to COMP:
Trichodesmium (Figure 14.7b) 56 m in diameter; (b) colonial, Pleurocapsa; these
BarnsBarns

H. Buckley

H. Buckley
structures contain hundreds of cells and are 7 50
Please put figure
SusanSusan

DanielDaniel

DanielDaniel
Figure 14.2 (c)
m in diameter; filamentous, Lyngbya;
Cyanobacteria: the five amajor
single
Group IV, Nostocales. Nodularia (Figures 14.2d ), Nostoc,

Daniel H. Buckley
cell measures
morphological
legend about
types 10ofspace
in m wide; (d) filamentous
cyanobacteria.
in
(a) (b) (c)
heterocystous, Nodularia; aasinglesinglecell
cellmeasures
measuresabout
Filamentous cells that
(a) produce (Figure 14.8ab),
Calothrix (b) (a)
(c)56
Unicellular, Gloeothece;
10 m note
mwide;
lower
in toleftCOMP:
(e) filamentous
diameter; (b)cornerbranching,
colonial, Fischerella;
Pleurocapsa; these
heterocysts Anabaena (Figure 14.6c), a cell is about
structures
diversity
m
Please
10 m
contain
relates to
in diameter;
wide. See
hundreds
put figure
(c)phylogenetic
how and
of cells
filamentous,diversity
morphological
are 7 50
Lyngbya;inaFigure
single14.3. (d) (e)
Figure 14.2 Cyanobacteria: the five major
Cylindrospermum, Scytonema,

Daniel H. Buckley
cell measures about 10
legend in space in m wide; (d) filamentous
morphological
Figure 14.2 types of cyanobacteria.
Cyanobacteria: the five major heterocystous, Nodularia; a single cell measures about
(a) Unicellular,
morphological
note types of cyanobacteria.Richelia (Figure 14.7c)
Gloeothece;
to COMP:
a single cell measures
10 m wide;
lower(e) filamentous
left corner branching, Fischerella;
56 m in diameter; (b) colonial, Pleurocapsa; these
(a) Unicellular, Gloeothece; a single cell measures a cell is about 10 m wide. See how morphological
structures note
containtohundreds
COMP: of cells and are 7 these
50
Group V, Stigonematales. Cells 56 m Please
m in
in diameter;
diameter; (c)
(b) colonial,
put figure
filamentous, Fischerella (Figures 14.2e, 14.8c, d ),
Pleurocapsa;
Lyngbya;
structures contain hundreds of cells and are 7 50 a single
diversity relates to phylogenetic diversity in Figure 14.3. (d) (e)
Please put mfigure

H. Buckley

H. Buckley
cell measures about 10 space wide;Lyngbya;
(d)
in filamentous
divide to form branches m in legend
diameter;
heterocystous,
in
(c) filamentous,
Nodularia; a single Stigonema, Chlorogloeopsis,
cell
a single
measures about

H. Buckley

H. Buckley
cell measures
legend about 10 m
in corner
spacewide; (d) filamentous
in Fischerella;
10 m wide;
lower(e)Nodularia;
filamentous
left Hapalosiphon
branching,
anielDaniel

anielDaniel
heterocystous, a single cell measures about
a10cell is about
m wide; 10 m wide.
(e) filamentous See how
branching,morphological
Fischerella;
lower left corner
diversity relates to phylogenetic diversity in Figure 14.3. (d) (e)
Dendrograma que representa relaciones filogenticas inferidas a partir del anlisis de familias de protenas
conservadas en genomas de cianobacterias. Los crculos de colores se utilizan para indicar los rasgos de
las especies segn lo indicado por la clave.
 3,5 Billones de aos

Bacterias Gram-negativas

Clorofila a b o d ; ficobiliprotenas; glucgeno; pared celular aminoazcares

y aa

ADN cianobacterias unicelulares 1,6 x 109 8,6 x 109 Dalton (1,0 x 109
3,6 x 109)
Plate 1. Diagram showing the major respiratory and photosynthetic electron transport components of cyan
ails, see Chapters 613.
Protoplasma perifrico tilacoides & ficobilisomas

Diagrama de los principales componentes de transporte de electrones respiratorios y forosintticos de

cianobacterias
 Ribosomas 70S

Cianoficina polmero proteico no ribosmico grnulos en el citoplasma

CYANOBACTERIA 39
(cantidades equimolares de arginina y cido asprtico)

Microfotografa electrnica de transmisin de una seccin de una clula de Plectonema

Fig. 2.11 Drawing of the fine-structural features of a
boryanum que muestras cuerpos de cianoficina (C).
cyanobacterial cell. (C) Cyanophycin body (structured
granule); (Car) carboxysome (polyhedral body); (D) DNA
fibrils; (G) gas vesicles; (P) plasmalemma; (PB) polyphosphate
 C-ficocianina [] 620 nm

Aloficocianina [max] 650 nm

C-ficoeritrina [max] 565 nm

Ficoeritrina [max] 568 nm

 Acinetos clulas ms grandes que las clulas vegetativas concentracin de
glucgeno & cianoficina

No son metablicamente inactivos; ni resistentes a las condiciones ambientales

extremas pierden el 90% de su capacidad fotosinttica & respiratoria

Solo en cianobacterias que forman heterocistos

Diferenciacin en acinetos deficiencia en fosfato; C; limitacin de C;

reduccin de la disponibilidad de energa lumnica
 Heterocistos clulas ms grandes que las clulas vegetativas
fotosintticamente inactivos

No fijan CO2; no producen O2; tasa respiratoria (consumo de O2); pared celular
gruesa y laminada limita ingreso de gases O2

Entorno interior virtualmente anxico

Nitrogenasa sensible al O2
 CYANOBACTERIA 41

Fig. 2.16 Light micrograph of Anabaena crassa showing

Microfotografa de and
vegetative cells, akinetes, Anabaena crassa
heterocysts. (From mostrando
Li et al., clulas vegetativas, acinetos y heterocistos
1997.) the gas vesicle is quite rigid, with the gas inside
it at a pressure of 1 atm. The membrane is per-
meable to gases, allowing the contained gas to
glucose molecules, that is similar to amylopectin equilibrate with gases in the surrounding solu-
 Algunas especies diazotrficas (capaces de fijar el N2)
CYANOBACTERIA 49

Qumica de la fijacin de nitrgeno y la subsecuente

incorporacin del nitrgeno fijado en glutamato y glutamina
Fig. 2.27 The chemistry of nitrogen fixation and the
subsequent incorporation of the fixed nitrogen into