LECTURE-6

BACTERIA
 Bacteria are procaryotes.
 They are small organisms generally enclosed by rigid cell walls.
 The outer surface of the cell wall in many bacterial species are
covered with a slimy, gummy coating known as a capsule or slim
layer.
 Bacterial species are unicellular.
 They may exist in three basic morphological forms, i.e. cocci
(spheres), bacilli (rods), spiralli (Sptiral) as reflected in the following
Figure-1:

Figure-1: Various types of bacteria.

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 Most cannot utilize light energy are capable of motion (motile), and
reproduce by division into two daughter cells (binary fission).
 There are many subdivisions of bacteria: some of the general groups
of bacteria are given in Table-1.
 Their typical properties are also given in the Table.
Table-1: Some Major Groups of Bacterial Species and their
Distinguishing Characteristics

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 The column levelled "Gram reaction" refers to the response of the
bacteria to the staining test.
 Cells are first stained with the dye crystal voilet, then treated with
an iodine solution and washed in alcohol as shown in Figure-2.

Figure-2: Gram staining technique.

Figure-3: Gram positive vs gram negative bacteria.

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  The solubility of oxygen in the medium has major implications in
bioprocess design.
 In anaerobic process such as production of alcohol and digestion of
organic wastes don't require oxygen.
 The bacteria may form endospores (dormant form) under adverse
condition during commercial application.
 Spores are the dormant form of the cells capable of resisting heat,
radiation and poisonous chemicals.
 When the spores are exposed to the suitable environment in terms of
nutrients, temperature, pH etc., they start giving normal functions.
 The normal biologically active cells are the vegetative form of cells
which distinguish it from the spore form.
 As shown in Table-1, there are two major groups of spore forming
bacteria.
 The aerobic bacillus species are widespread and adaptable.
 Many clostridium species which function normally under anaerobic
conditions die in the presence of oxygen in the vegetative state but
form spores unaffected by oxygen.
 Some bacteria whose vegetative forms are rapidly killed at 45 0C
can form spores which survive boiling in water for several hours.
 Therefore, when we are trying to kill bacteria by heat sterilization
the spore forming capability of bacteria need higher temperature to
be killed typically under high pressure of steam i.e. 15 psig in an
autoclave at temperature more than 1210C.

YEAST
 Yeasts are unicellular eukaryotes that belong to the Kingdom of Fungi
 They are one of the important subgroups of fungi.

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 Like bacteria, yeasts are widespread in nature.
 Yeasts are found in water, soil, and plant leaves.
 They are also found on animal skin or inside an animal symbiotically.
 They are usually found in regions of lower relative humidity than
bacteria.
 They cannot extract energy from sunlight and are usually free-living.
 Although most fungi have complex morphology, yeasts are
distinguished by their usual existence as single, small cells from 5 to
30 µm long and from 1 to 5 µm wide.
 With over 1,500 species identified so far.
 However, some yeast-like fungal parasites, such as Candida albicans,
could cause oral and genital infections in humans.
 Yeast is one of the earliest domesticated organisms.
 Saccharomyces cerevisiae is the most commercially significant species
that has been used by humans to produce bread, wine and beer for
hundreds, if not thousands of years.
 Some images of the yeast cells have been shown in the following
Figure-4:

Figure-4: Images of yeast cells.

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 YEAST GROWTH
 Yeasts are non-pathogenic and are very easy to work with.
 Yeasts can be grown in chemical defined media both in liquid
culture and on solid agar.
 Both Saccharomyces cerevisiae and Schizosaccharomyces pombe can
grow optimally at 30°C with an average doubling time of 90 – 120
minutes.
 However, they also have the ability to grow at temperature as low as
16°C and as high as 42°C.
 In between usage, yeast can be stored in 10% glycerol at -80°C
freezer for years.
 THE YEAST LIFE CYCLE (REPRODUCTION OF YEASTS)
 Various paths of reproduction of yeasts are asexual (budding and
fission) as shown in Figure-5 and sexual.
 In budding a small offspring cell begins to grow on the side of the
original cell.

Figure-5: Reproduction of yeast cells.

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 Physical separation of matured offspring from the parent may not
be immediate, and formation of clumps of yeast cells involving
several generations are possible.
 Reproduction by budding has been shown by Figure-6 and Figure-7.

Figure-6

Figure-7

 Fission occurs by division of the parent cell into two new cells as
shown in above in Figure-8.

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  Sexual reproduction occurs by conjugation as showb in Figure-5.
 It occurs by conjugation of two haploid cells (each having a single
set of chromosomes) with dissolution of the adjoining walls to form
diploidcell (two sets of chromosomes per cell) zygote.
 The nucleus in the diploid cell may undergo one or several divisions
and form ascopores
 Each of these eventually becomes an individual new haploid cell.
 Which may then go subsequent reproduction by budding, fission, or
sexual fusion again.

MOLDS
 Mold are higher fungi with a vegetative structure called a mycelium.
 As shown in Figure-10, the mycelium is a highly branched system of
tubes
 Within these enclosing tubes is a mobile mass of cytoplasm containing
many nuclei.
 The mycelium may consist of more than one cell of related types.
 The long thin filaments of cell within the mycelium is called hyphae.

Figure-10

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 In some cases the mycelium may be very dense.
 This property, may create mass-transfer resistance problem during
grwth and cultivation.
 Since the normal function is effected due to mass transfer resistance,
the common exponential growth law may not be followed.
 Molds, like yeasts, do not contain chlorophyll, and they are generally
non-motile.
 Reproduction may be sexual or asexual, accomplished means of
spores.
 Spores propertiesform an important comp-onent in fungi
classification.
 The commercially most important forms of molds are Spergillus and
Penicillium as demonstrated by the Figure-11.

Figure-11: Spergillus and Penicillium.

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 The major useful products of these organisms include antibiotic,
organic acids, and biological catalysts.
 The strain Aspergillus niger normally produces oxalic acid
(HO2CCO2H).
 Limitation of both phosphate nutrient and certain metals such as
copper, iron, and manganese results in a predominant yield of citric
acid.
 This limitation method is the basis for the commercial production of
citric acid by biotechnological method.
 A. niger is an interesting example differentiating approches to
biochemical reactor design and optimization from those of chemical
reactors.

ALGAE
 Algae are a diverse group of aquatic organisms that have the ability to
conduct photosynthesis.
 Certain algae are familiar to most people; for instance, seaweeds (such
as kelp or phytoplankton), pond scum or the algal blooms in lakes.
 However, there exists a vast and varied world of algae that are not
only helpful to us, but are critical to our existence.
 The term "algae" covers many different organisms capable of
producing oxygen through photosynthesis (the process of harvesting
light energy from the sun to generate carbohydrates).
 These organisms are not necessarily closely related.
 However, certain features unite them, while distinguishing them from
the other major group of photosynthetic organisms: the land plants.
 Primarily, algae are not highly differentiated in the way that plants
are.

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 That is to say, they lack true roots, stems and leaves, and a vascular
system to circulate water and nutrients throughout their bodies.
 Second, many algae are unicellular, according to a 2014 article
published in the journal Current Biology.
 They also occur in a variety of forms and sizes.
 They can exist as single, microscopic cells; they can be macroscopic
and multicellular; live in colonies; or take on a leafy appearance as in
the case of seaweeds such as giant kelp.
 Picoplankton are between 0.2 to 2 micrometers in diameter, while the
fronds of giant kelp are as large as 60 meters in length.
 Lastly, algae are found in a range of aquatic habitats, both freshwater
and saltwater.
 By virtue of these characteristics, the general term "algae" includes
prokaryotic organisms — cyanobacteria, also known as blue-green
algae — as well as eukaryotic organisms (all other algal species).
 "Since "algae" do not form a natural group that has descended from a
common ancestor, including cyanobacteria into the informal group
"algae" is common," said Linda Graham, a professor of botany at the
University of Wisconsin-Madison.
 "The term 'eukaryotic algae' excludes cyanobacteria."
 It is also interesting to note that chloroplasts, which are the site of
photosynthesis in land plants, are adapted forms of cyanobacteria.
 These early cyanobacteria were engulfed by primitive plants cells
sometime in the late Proterozoic, or in the early Cambrian period,
according to the University of California Museum of Paleontology.

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 Blue Green Algae
 Blue-green algae (Figure-12), also called cyanobacteria, any of a
large, heterogeneous group of prokaryotic, principally
photosynthetic organisms.
 Cyanobacteria resemble the eukaryotic algae in many ways,
including morphological characteristics and ecological niches, and
were at one time treated as algae, hence the common name of blue-
green algae.
 Algae have since been reclassified as protists, and the prokaryotic
nature of the blue-green algae has caused them to be classified
with bacteria in the prokaryotic kingdom Monera.

Figure-12: Blue-Green Algae

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 Significance of Algae
 Probably the most important contribution of algae to our
environment and well-being is the generation of oxygen through
photosynthesis.
 "Algae are indispensable because they produce about half the
oxygen in Earth's atmosphere," Graham told LiveScience.
 According to a 2010 review article published in the journal Biofuels,
petroleum is partially derived from ancient algae deposits.
 Algal biofuels are a promising replacement for fossil fuels.
 All algae have the ability to produce energy-rich oils and several
microalgal species naturally accumulate high levels of oil in their
dry mass.
 Moreover, algae are found in diverse habitats and can reproduce
quickly.
 They also efficiently use carbon dioxide.
 "Algae help to keep atmospheric carbon dioxide levels stable by
storing [the gas] in organic materials that include oil deposits and
inorganic carbonate rocks," Graham said.
 Green algae, diatoms and cyanobacteria are just some of the
microalgal species that are considered good candidates for the
production of biofuel (Biofuels, 2010).
 Algae may hold potential as both the food and the fuel of the future.
 The type of microalgae which may be used as food and fuel have
been shown in Figure-13.
 Single Cell Proteins (SCP) produced from microalgae have been
shown in figures-

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Figure-13: Microalgae which may be used as food and fuel.

Figure-14: SCP Processing Unit using algae

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 Figure-15: Spirullina Powder (SCP).

Figure-16: Second Generation (SCP) Product in the form of Tablet.

 ALGAL BLOOMS
 Algae, in the form of algal blooms, get a bad rap for creating toxic
conditions in oceans and lakes.
 "Algal blooms" refers to the rampant growth of certain microalgae,
which in turn leads to the production of toxins, disruption of the
natural aquatic ecosystems and increases the costs of water
treatments, according to the Environmental Protection Agency
(EPA), USA.
 The blooms take on the colors of the algae contained within them.

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  It is stated that the main toxin producers in oceans are certain
dinoflagellates and diatoms.
 In freshwaters, cyanobacteria are the main toxin producers, though
some eukaryotic algae also cause problems.
 Under natural conditions, algae use the toxins to protect themselves
from being eaten by small animals and only need a small amount to
protect themselves.
 The main cause of algal blooms is a phenomenon called nutrient
pollution.
 With nutrient pollution, there is an excess of nitrogen and
phosphorus, which can push algae toward unrestrained growth.
 The phenomenon is caused by a variety of human activities.
 The fertilizers we use in agriculture and animal manures are rich in
nitrogen, while improperly treated wastewater is high in both
nitrogen and phosphorus, according to the EPA.
 "It is a common societal perception that algae are noxious and
should be eliminated at every opportunity. But that perception is
wrong, because algae make oxygen, fish [they are a major source of
food for aquatic organisms], oil, and many other useful materials

PROTOZOA
 Protozoa are single celled organisms.
 The Protozoa are considered to be a subkingdom of the kingdom
Protista, although in the classical system they were placed in the
kingdom Animalia.
 More than 50,000 species have been described, most of which are free-
living organisms; protozoa are found in almost every possible habitat.

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  They come in many different shapes and sizes ranging from
an Amoeba which can change its shape to Paramecium with its fixed
shape and complex structure.
 They live in a wide variety of moist habitats including fresh water,
marine environments and the soil.
 Some are parasitic, which means they live in other plants and animals
including humans, where they cause disease.
 Plasmodium, for example, causes malaria.
 They are motile and can move by:
 Cilia - tiny hair like structures that cover the outside of the
microbe. They beat in a regular continuous pattern like flexible
oars.
 Flagella - long thread-like structures that extend from the cell
surface. The flagella move in a whip-like motion that produces
waves that propel the microbe around.
 Amoeboid movement - the organism moves by sending out
pseudopodia, temporary protrusions that fill with cytoplasm
that flows from the body of the cell.
Structure
Protozoa are microscopic unicellular eukaryotes that have a relatively
complex internal structure and carry out complex metabolic activities.
Some protozoa have structures for propulsion or other types of movement.

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Figure-18: Fine structure of a protozoan parasite, Typanosoma evansi, as
revealed by transmission electron microcopy of thin sections.

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Classification
On the basis of light and electron microscopic morphology, the protozoa
are currently classified into six phyla. Most species causing human disease
are members of the phyla Sacromastigophora and Apicomplexa.
Life Cycle Stages
The stages of parasitic protozoa that actively feed and multiply are
frequently called trophozoites; in some protozoa, other terms are used for
these stages. Cysts are stages with a protective membrane or thickened
wall. Protozoan cysts that must survive outside the host usually have more
resistant walls than cysts that form in tissues.
Reproduction
Binary fission, the most common form of reproduction, is asexual; multiple
asexual division occurs in some forms. Both sexual and asexual
reproduction occur in the Apicomplexa.
Nutrition
All parasitic protozoa require preformed organic substances—that is,
nutrition is holozoic as in higher animals.

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