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Basics Concepts of

Manufacturing II
Prof.: Jacqueline Hernández
Estudiar ciertos aspectos de la
microbiología enfocados desde el
punto de vista industrial.
Se hará énfasis en la importancia y
utilidad de algunos microorganismos
en procesos industriales.

 The study of  Fungi - single &
microscopic multi cell forms -
organisms which yeast, 
include many life- filamentous
forms: molds,  complex
 Bacteria - simple, fungi
single cell 

 Protists - single cells,  Viruses - acellular, 
some multicellular  protein-based
algae, protozoans,  lifeforms,
slime molds typically
 intracellular

 Microbiology is not a new field of study.
 In 1674 Anton van Leeuwenhoek  used
simple, self-made  microscopes to examine
stagnant H2O, where he observed "wee
animalcules"  - microscopic organisms. 
 Animalcules“ - microscopic organisms. 

 Hisdetailed descriptions and drawings of
these microscopic forms of life were
published by the Royal Society of
 Many others also made significant
contributions including Louis Pasteur,
Edward Jenner and Robert Koch as a few

Areas of Microbiology

 Microbiology can be examined in a number of

ways, although focusing on the nature of
specific micro-organisms or on health-
related aspects are the most common.
 Organization by Organisms
 Bacteriology – study of bacteria
 Virology  - study of viruses
 Parasitology – study of parasites
 Mycology – study of fungi
 Phycology - study of algae
 Protozoology – study of protozoa

Organization by Health Science

 Epidemiology – study of disease patterns in

 Etiology – identification of agents which cause

 Immunology – study of immunological

responses to micro-organisms
 Infection control – control of spread of

infectious disease

Areas of Study and Research

 In addition to being causative agents of

infectious diseases there are many other
aspects of microbiology which need to be
 As normal flora in humans (or other living
systems) there are significant benefits with
respect to metabolic functions as well as
preventing invasion by pathogens (positive
antagonistic effects).

Areas of Study and Research
 In the environment micro-organisms serve
many beneficial roles in the food chain, as
decomposers, in sewer / wastewater
treatment, etc.
 In industry micro-organisms are essential
components of the food production, brewing
/ beer production, pharmaceutical
industries and many other areas.

Areas of Study and Research
 Micro-organisms survive in every possible
environment, including those that are
considerable inhospitable.
 The air we breathe, the water, soil, our skin
surface, our intestines - all harbor thousands
of microscopic organisms.

Areas of Study and Research
 There are a number of ways of classifying
 The (2) Fundamental Cell Types
 Eukaryotic  (“true nucleus”)  -  more complex
 Prokaryotic  (“pre-nucleus”)  -  simpler,

Una comparación de los sistemas de clasificación en reinos biológicos más notables:

Comparación en Reinos
Haeckel (1894) Whittaker (1969)&sup1 Woese (1977) Woese (1990)
Tres reinos Cinco reinos Seis reinos Tres dominios

Protista Monera Eubacteria Bacteria

Archaebacteria Archaea

Protista Protista Eukarya

Plantae Fungi Fungi

Plantae Plantae

Animalia Animalia Animalia

Eukaryotic Cells
Eukaryotic Cells
 1. Nucleus - eukaryotes have a "true nucleus”.
 Genetic / nuclear material surrounded by a
nuclear membrane.
 Genetic / nuclear material organized into paired
 Nuclear membrane (DNA) associated with
proteins called histones.
 Nucleus contains nucleolus - sites of ribosome
 2. Complexity - internal structure more complex –
 has "organelles"
 3. Cell walls
 Found only in plant cells, fungi
 Composed of cellulose, chitin.

 4. Replication / division occurs by mitosis, meiosis.

Prokaryotic Cells
Prokaryotic Cells

 1.Nucleus - no “true” nucleus.

 lack a nuclear membrane.
 lack histones.

 lack nucleolus.

  2.   Complexity - No organelles.
  3.   Cell walls
 All typical prokaryotic cells possess cell
 Cell walls made of a peptidoglycan layer.

 4.   Replication / division - binary fission.

Examples Micro-organisms of each cell
1. Eukaryotes - fungi, protozoa,
 algae
2. Prokaryotic - bacteria
3. Viruses - belongs in neither group
- (acellular, protein-RNA based

Growth of Micro-organisms

 1.Reproduction - Generation Times

 Doubling time - time required for one cell
to produce two new cells.
 Varies with type, age and health of 

organism and environmental conditions.

 Can range from 15 min. to as much as 24

 Population constantly doubles - increases


Growth of Micro-organisms
 2.   Measuring Bacterial Growth
 Plate count (standard plate counts)
 Direct microscopic count
 Turbidity / optical density

Growth of Micro-organisms
 3.  Phases of
 Micro-organisms
move through
four stages of
growth including
 lag, logarithmic,
stationary and
death phases

Growth of Micro-organisms
 Lag Phase:
 increased metabolic activity
 no increased  number of cells
 cells are preparing for reproduction and
accumulating nutrients and energy
 Exponential / Logarithmic Growth Phase
(Log Phase)
 microbial population is constantly doubling -
rapid growth / reproduction
 diagnostic testing and  antibiotic therapies most
effective in this phase

Growth of Micro-organisms
Stationary Phase
population numbers are static: rate of
reproduction = rate of death
occurs due to exhaustion of nutrients,
accumulation of wastes, etc..
Death Phase
rate of death higher than rate of
rate of death is species-variable

Growth of Micro-organisms
Nutrients - fundamental nutritional
requirements include Carbon, 
Nitrogen, Hydrogen, Oxygen as well as
trace elements and minerals. 
There may be a requirement for special
growth factors for more fastidious
 Theserequirements could include
vitamins, amino acids, carbohydrates,
blood factors

Parameters Influencing Bacterial Growth

 Microbial populations are both extraordinarily

adaptable and yet can be remarkably
sensitive to "rate limiting" environmental
conditions. Example factors which may
influence growth can include:

Parameters Influencing Bacterial
 Temperature -  all bacteria  have an
optimum temperature range.
 Based on temperature sensitivity there are
three categories of micro-organism: 
 Mesophiles (20°C - 50°C)
 Thermophiles (50° - 80°C) 
 Hyperthermophiles) - up to 110°C
 Psychrophiles  (0°C  - 20°C)

Parameters Influencing Bacterial
 Classification by Nutrient
Autotrophic - independent -
require light, inorganic
dependant - require organic

Parameters Influencing Bacterial
 Oxygen and other Gases - there are
groups of micro-organisms based on
oxygen needs:
 Aerobic - require gaseous molecular oxygen
 Anaerobic - do not require gaseous oxygen,
acquire chemically
 Facultative - aerobes or anaerobes as
environment dictates

Parameters Influencing Bacterial
Moisture - each organism has a
minimum requirement for water.
In addition to forming the cytosol,
water levels affect osmotic pressure
(the pressure exerted by
concentration of solutes in water). 
This osmotic pressure can have a
profound impact on most organisms.
Parameters Influencing Bacterial
 pH -  most micro-organisms have a range of
pH values over which they function well.
 With acidic or basic conditions outside of this
range there can be a significant drop in
survival as enzymatic systems and
metabolism shuts down.
 Acidophiles prefer a range of pH 0-5,
 Neutrophiles  ~ 7.0, and
 Alkalinophiles pH 8-12.

Parameters Influencing Bacterial
Salinity - closely related to
moisture requirements and
osmotic pressure.
Some micro-organisms can survive
much higher levels of salt.
Halophillic organisms require 3%
salt but may range as high as 15-
30% salt levels.

Nomenclature and Microbial Taxonomy

 One of the most cited taxonomic

references for microbial classification is 
Bergey’s Manual (of Systematic
Bacteriology) which describes
physiological characteristics and
chemical testing.   
 In general, the prokaryotes are classified
using the following taxonomic groups:
 Division -- Class -- Order -- Family --
Genera -- Species -- Strain

Techniques used to Classify
 Bacteria represent a significant source of
pathogenic infections in humans.
 There are many ways to classify bacteria.
Examples include:
 cell morphology - size and shape (rods,
spheres, spiral)
 colony shape (on petri plates)
 stain uptake (use of characterizing stains- i.e.
Gram stains)

Techniques used to Classify
 motility (mobile / immobile)
 energy sources (autotrophs, heterotroph, etc.)
 growth requirements (pH, salinity , oxygen,
 products (fermentation / growth products)
 antibiotic response and sensitivities
 Cell wall composition.
 Nucleic acid  profiling (sequence comparisons)
 Serotyping and immunological testing
 RFLP analysis and genetic hybridization
Enumeration of Cells
 Numbers of microorganisms can be estimated
based using a number of techniques which
include conducting representative cell
counts (on plates or other media).
 Direct microscopic counts are conducted by
examining a known volume of cell suspension
under a microscope.
 Direct microscopic counts enumerate living and
dead cells equally.
 The use of stains (i.e. Gram stain, etc.) can
increase visibility and allow for both
qualitative and quantitative assessments.

Streak PLate
Standard Plate Count
 The SPC (standard plate count) is still the
single most common method of
enumerating cells.
 A precise volume of liquid (media /
sample) containing the cells is placed on
the surface of an agar-containing petri

Standard Plate Count
 The plate is incubated (typically >24 hours)
and the plate is counted for colonies.
 Each colony is assumed to be derived from a
single bacterial cell - allowing the
microbiologist to relate the number of
colonies (CFU - colony forming unit) and
dilution factors to determine the original
number of organisms in the sample.
 This is usually expressed as CFU / ml (colony
forming units per milliliter).

Serial dilution
Serial Dilutions
 Alternatively, changes in turbidity can be
measured to assess growth of bacteria in
clear medias / broth. 
 This can be conducted using a
spectrophotometer or a tubidometer and
provides rapid data.
 Optical techniques however, do not
typically differentiate between live and
dead cells.

 Amor Fraterno:
 Si nos amamos
unos a otros , Dios
vive en nosotros ”.
 1Juan - 4 ; 12