2. Describe and differentiate between prokaryotic, archaea and eukaryotes cells.

Description:

Prokaryotic cells are those that do not have a differentiated nucleus, so their DNA is located in the
cytoplasm, but not enclosed in a membranous covering as is the case with eukaryotic cells. In
addition, they contain cell membrane, cell wall, cytoplasm, and ribosomes. Virtually all prokaryotic
cells are unicellular organisms, but like all exceptions, there are also multicellular organisms in this
type of cell.

Prokaryotic organisms are evolutionarily prior to eukaryotes. And although prokaryotic cells arose
in a very remote past, that does not mean that they have disappeared from Earth. In fact, the
simplest life forms are still prokaryotic organisms, like bacteria.

A eukaryotic cell is considered to be all those cells in whose cytoplasms a well-defined cell nucleus
can be found, which contains most of its genetic material (DNA), eukaryotic cells have organelles
or organelles, which are special subcellular structures that can They are identified within and are
delimited by membranes such as mitochondria and ribosomes.

The emergence of eukaryotic cells signals an important step in the evolution of life, as it laid the
foundations for much greater biological diversity, including the emergence of specified cells within
multicellular organizations, originating kingdoms: protists, fungi, plants, And animals.

Archaea are a group of unicellular organisms similar to bacteria, but so different from them that
they are classified in their own taxonomic kingdom. These were initially classified as bacteria
because both bacteria and archaea lack a cell nucleus, as well as the majority of organelles present
in eukaryotic cells. But the metabolic differences between the two groups are so great that they
ended up being classified into different domains. The most striking characteristic of archaea is that
many live in extreme environmental conditions, such as salty lakes or hot water fumaroles on the
ocean floor. The organisms that inhabit these conditions are known as Extremophiles.

Differences:

Eukaryotic cells in comparison with archaea and prokaryotic cells show a great difference as they
are mostly multicellular organisms, having a defined nucleus, being of a larger size and being more
complex cells with organelles performing specific functions within them. These also differ in
reproduction because eukaryotic cells in addition to asexual reproduction (Prokaryotes by binary
fission, eukaryotes by mitosis), as well as sexual reproduction by meiosis.

Among the differences between prokaryotes and archaea, apart from resistance to antibiotics, are
the structure of the cell membrane, the size of its ribosomes and its flagella. The cell membrane of
archaea is different from that of bacteria and eukaryotes, having a unique chemical composition,
made up of long chains (from 20 to 25 carbons) and branched isoprenoids, which are joined at
each end by ether bonds to a glycerol, which in this case has a phosphate group on the first
carbon. This type of phospholipid forms a lipid monolayer. Their flagella (organelles that
unicellular organisms use to move around in liquid media) are also different. Unlike bacteria, the
cell wall of archaea does not contain peptidoglycans and is made up of proteins, polysaccharides
or glycoproteins. Some archaea have a pseudopeptidoglycan with different sugars in the
polysaccharide.

4. What type of cells have a nucleus? Nucleoid? What is a cell’s genome and why is it important?

Cells with a nucleus defined as such are eukaryotic cells, which is made up of a nuclear membrane
that delimits with the rest of the cytoplasm and in which the majority of genetic material is found,
on the other hand, cells that have nucleoids are prokaryotes, which is not a defined or delimited
sector, but an area within the cytoplasm where the genetic material is concentrated, it should be
noted that it does not have a membrane.

A genome is an organism’s complete set of DNA, including all of its genes. Each genome contains
all of the information needed to build and maintain that organism. In humans, a copy of the entire
genome—more than 3 billion DNA base pairs—is contained in all cells that have a nucleus.

It is one of the most important advances in science, it is so important that thanks to it we have
been able to identify the functions of all our proteins. These are, ultimately, those that define
normality and alterations (pathologies) of our biological functions. Human medicine and
immunology have already begun to benefit from these advances.

6. Name and describe the three domains of life.

Archaea: Organisms in the Archaea domain are unicellular and do not have a nucleus or organelles
attached to the membrane. Despite their similarity to prokaryotes in the Bacteria domain, they
have a different biochemistry that distinguishes them. Between these they present so much
difference in their function and structure that it is considered that it did not make sense to be
grouped into one. Organisms that belong to this domain are Extremophiles, organisms that thrive
in high salinity, temperature, or acidity, and methanogens that produce methane.

Prokaryotic: This comprises the majority of microscopic unicellular living beings. They are
characterized mainly by not having a nucleus or organelles, they can also have a peptidoglycan cell
wall. They are considered the most abundant organisms on the planet and are found in every
corner of the globe, even with extreme conditions where other living beings would not be able to
survive.

Eukarya: Este reino está gobernado por las eucariotas y se pueden considerar como los
organismos más evolucionados según sus características. Se caracterizan por ser células con
núcleo y orgánulos más complejos como las mitocondrias o los cloroplastos, con complejos
mecanismos bioquímicos que puede otorgarles la posibilidad de abandonar la condición de
unicelulares y empezar a formar complejos sistemas pluricelulares.

8. What advantages do solid media offer for the isolation of microorganisms?

The advantage of solid culture media is that it allows detecting the different types of bacteria that
can be found in a single sample, then using one or the other culture medium depending on what
we are looking for, although it is necessary to isolate all the bacteria that there may be in a sample
the most recommended is or would be to use a solid culture medium.

They are used to obtain bacteria isolated by the formation of colonies on the surface of the culture
medium and for the study of the morphology of the colonies, which liquid media or broths do not
allow.

10. What is meant by the term “enrichment culture”?

Enrichment culture is an isolation technique that was designed to make growth conditions
favorable for an organism of interest and have an unfavorable environment for any competition. It
is also defined as the use of certain growth media to promote the growth of a particular
microorganism over others. This enrichment is generally done by introducing nutrients or
environmental conditions that only allow the growth of an organism of interest. Enrichment
culture techniques are used to increase a small amount of the desired organisms to detectable
levels.

12. What color will a gram-negative cell be after Gram staining by the conventional method?

After staining, the gram-negative cells will stain pink, and this is because the wall of the Gram-
negative bacteria owes its rigidity to a thinner layer of murein, through which the alcohol extracts
the crystal violet from these cells. Its cytoplasm is colorless and can be stained pink or red with the
contrast dye: safranin

14. What characteristic forms can bacteria take? Describe the ways that bacterial cells can group

You can see three different ways. These can be identified and classified by their shape, from this
classification 3 groups stand out, which are the bacilli, which are rod-shaped, the cocci, which are
sphere-shaped, and spirilla, which are spiral-shaped.

16. Why do bacterial cells need cell walls? Do all bacteria have cell walls?

The bacterial cell wall is necessary for bacteria because it performs different functions in addition
to providing general strength to the cell. It also helps maintain the shape of the cell, which is
important for knowing how to grow, reproduce, mean nutrients, and move. It also protects the
cell from osmotic lysis, as the cell moves from one environment to another or carries nutrients
from its environment. Since water can move freely through the cell membrane and cell wall, the
cell is at risk of an osmotic imbalance, which could put pressure on the relatively weak plasma
membrane.

No, not all bacteria have a cell wall. However, it is important to note that most bacteria (about
90%) have a cell wall and usually have one of two types: a gram positive cell wall or a gram
negative cell wall.

Bacteria that lack a cell wall completely makes them extremely susceptible to osmotic changes.
They often strengthen their cell membrane somewhat by adding sterols, a substance generally
associated with eukaryotic cell membranes. Many members of this phylum are pathogenic and
choose to hide within the protective environment of a host.
18. Why does alcohol readily decolorize gram-negative but not gram-positive bacteria?

When alcohol (ethanol, in most stains) is added to a mixture of Gram-positive and Gram-negative
cells, the Gram-positive cells are not affected because they cannot pass through the thick layer of
murein (they can if they are excessively prolonging contact with alcohol). Instead, the wall of Gram
negative bacteria owes its stiffness to a thinner murein layer, through which alcohol has an easier
time entering the cytoplasm and discoloring the bacteria.

20. How do fimbriae differ from pili, both structurally and functionally?

The fimbriae and pili are filamentous structures composed of proteins that extend from the
surface of a cell. Fimbriae are found in gram-negative and gram-positive bacteria, and pili only in
gram-negative bacteria.

Fimbriae are small bristle-shaped fibers that emerge from the surface of bacterial cells while pili
are microfiber-like hairs that have a thick tubular structure formed by pilin, in terms of general
characteristics, pili are larger, thicker and rigid, but they are found to a lesser extent than the
fimbriae. Pili are made up of plasmid genes and their main function is responsible for bacterial
conjugation. Whereas the fibers are governed by bacterial genes in the nucleoid region and their
function is to be responsible for the attachment of the cell to the surface, and to allow the cell to
adhere to the surfaces of other bacteria.

22. What is meant by the word sterile? Why is aseptic technique necessary for successful
cultivation of pure cultures in the laboratory?

Sterility can be defined as the absence of the presence of viable microorganisms. Sterile conditions
require the complete absence of microorganisms, including bacteria, fungi, and their spores.

The aseptic technique is necessary for the successful cultivation of pure cultures because they
avoid contamination of the culture by external agents, thus ensuring that the culture remains
pure. If the aseptic technique is not carried out, it cannot be assured or it cannot be certain that a
pure culture was successfully re-performed.

24. Which nitrogen bases are purines and which are pyrimidines?

In the case of DNA, the bases are two Purines and two Pyrimidines. The purines are A (Adenine)
and G (Guanine). The pyrimidines are T (Thymine) and C (Cytosine). In the case of RNA there are
also four bases, two purines and two pyrimidines. The purines are A and G and the pyrimidines are
C and U (Uracil).

26. Describe the Gram stain method and how it works.

Gram stain is a common technique used to differentiate groups of bacteria based on their different
components in the cell wall. The procedure works between Gram positive and Gram negative
groups by coloring these cells red or purple. Gram positive bacteria stain purple due to the
presence of a thick peptidoglycan layer on their cell walls, which retains the crystal violet with
which these cells are stained. Alternatively, Gram negative bacteria stain red, which is attributed
to a thinner peptidoglycan wall, which does not retain crystal violet during the decolorization
process.
Gram stain involves three processes: staining with a water-soluble dye called crystal violet,
discoloration, and counterstaining, usually with saphanin. Due to differences in the thickness of a
peptidoglycan layer in the cell membrane between Gram positive and Gram negative bacteria,
Gram positive bacteria (with a thicker peptidoglycan layer) retain the crystal violet staining during
the bleaching process, while Gram negative bacteria lose crystal violet staining and are instead
stained with safranin in the final staining process. The process consists of three steps:

• Cells are stained with crystal violet stain. Next, a solution of Gram's iodine (iodine and potassium
iodide) is added to form a complex between crystal violet and iodine. This complex is a larger
molecule than the original crystal violet stain and iodine and is insoluble in water.

• A decolorizer such as ethyl alcohol or acetone is added to the sample, which dehydrates the
peptidoglycan layer, shrinking and tightening it. The large crystal violet-iodine complex cannot
penetrate this tight peptidoglycan layer and is therefore trapped in the cell in Gram positive
bacteria. In contrast, the outer membrane of Gram negative bacteria degrades and the thinner
peptidoglycan layer of Gram negative cells cannot retain the crystal violet-iodine complex and
color is lost.

• A counterstain, such as weakly water-soluble safranin, is added to the sample and stained red.
Since safranin is lighter than crystal violet, it does not alter the purple coloration in Gram positive
cells. However, discolored Gram negative cells stain red.

28. Describe the following types of media and their uses: defined or synthetic, complex, general
purpose, enriched, selective and differential. Give an example of each

Complex media: They are prepared from animal tissues, and more rarely from plants. Its
composition is not exactly defined, and therefore it is not rigorously constant. This can have
certain drawbacks under experimental conditions, where reproducibility may not be exact. In
current practice these means give excellent results and are the most widely used. For example:
Yeast extract

General purpose: It is the medium where all kinds of microorganisms grow, except those that need
special conditions.

Synthetics: are means that have a qualitatively and quantitatively defined chemical composition.
They are generally used in research work. Example: Czapek Agar.

Enriched: They contain additional components (in addition to the basic ones) to allow the
development of demanding microorganisms, which would not grow in a general medium, for
example: Blood agar, an enriched medium in which blood supplements provide them with all the
basic nutrients . CLED agar.
Selective: They have a component that prevents the development of unwanted microorganisms.
This makes it easier for the microorganism to be cultivated. For example, MacConkey Agar
contains crystal violet, which inhibits the growth of gram-positive bacteria and fungi, facilitating
the growth of gram-negative bacteria.

Differentials: They contain substances that reveal some characteristic of the species or group of
microorganisms. For example, TSI Agar: Triple Sugar Ferrum (Ferrum Three Sugars)

30. How do DNA and RNA differ?

The main differences between deoxyribonucleic acid and ribonucleic acid are:

Location: DNA is found in the nucleus, with a small amount of DNA also present in the
mitochondria. Whereas RNA is formed in the nucleolus and then moves to specialized regions of
the cytoplasm based on the type of RNA formed.

Structure: DNA is a long ladder-shaped macromolecule that twists to form a double helix and RNA
is generally single-stranded.

Nucleotides: Each nucleotide of DNA contains one of four nitrogenous bases, A (adenine), G
(guanine), T (thymine), or C (cytosine), and RNA contains uracil instead of thymine.

Sugar: DNA contains deoxyribose sugar and RNA contains ribose sugar

Number: For a particular species, the number of DNA remains constant for each cell while the
amount of RNA can differ from cell to cell.

Limits: DNA cannot leave the nucleus, RNA leaves the nucleus (mRNA).

Function: DNA has the function of storing genetic information, directing protein synthesis,
determining the genetic coding, and RNA's function is to transfer genetic information from DNA to
proteins, transport it out of the nucleus, translate it into proteins.

Types: DNA in chromosomal DNA (nuclear DNA) and extrachromosomal (plasmid DNA, mt DNA, cll
DNA, etc.) and RNA in messenger RNA (mRNA) transfer RNA (tRNA) ribosomal RNA (rRNA) etc.