Group #4

Group Members: Date Performed: June 17, 2019

Rejuso, Alyssa Rae Date Submitted: June 21, 2019
Romero, Timothy
Ruiz, Kim
Sacro, Rico Rafael
Sanchez, Kristel
Santiago, Carl Vincent

Exercise No. 1
Morphology and Motility of Microorganisms

Abstract

Microorganisms are said to be the most diverse and most abundant life forms on Earth; being classified
into Bacteria, Algae, Fungi, Protozoans, or Viruses. Regardless of their size, they serve significant functions
in the biosphere--ranging from advantageous to unfavorable effects. Given this, it is important to develop
an understanding of the purpose and implications of these organisms. However, in doing so, the
fundamental biological properties must be first taken into account. Morphology and motility are two of
the most basic concepts to instigate the journey into comprehending the field of microbiology. For this
study, various specimens of Bacteria, Fungi, Algae, and Protozoans were observed. Subsequently, their
morphologies were distinctly identified and analyzed. Upon examining the samples under different
classifications, it was discerned that most of them exhibited the most common cell shapes (rod-, circular-
and spiral-shaped), however, there were still a few that displayed very distinct morphologies (e.g.
Rhizopus sp., and Giardia lamblia trophozoite). In terms of motility, the wet mount and hanging-drop
techniques were utilized. Based on the specimens, Escherichia coli and Staphylococcus aureus, utilized,
their type of motility were determined; having true motility for the former, and Brownian movement for
the latter. Grounded on the information gathered, it was concluded that these fundamental biological
properties are interrelated to the functions and adaptations these microorganisms possess.

Keywords: morphology, motility, bacterial morphology, Brownian movement, hanging-drop technique

I. Introduction
Generally, microbiology tackles the essence and nature of microorganisms⏤small biological life
entities that are difficult for the unaided human eye to see. These microbes are characterized by their
diversity in terms of form and function, and exist in all three domains of life (Bacteria, Archaea, and
Eukarya). Hence, being deemed as the most abundant life forms on Earth. Given this, these microscopic
organisms include bacteria, archaea, fungi, protozoa, parasitic worms (helminths), and viruses (Sattley
and Madigan 2015). Although distinguished as minute living bodies, these still differ in terms of various
biological properties (e.g. motility, morphology, physiology, genetics, etc).
Microbes are often identified based on their shape and arrangement⏤their morphology. Noting
that these are the immediate characteristics revealed about microorganisms when microscopically
observed, it is apparent that morphology plays an important role in distinguishing various microbial cells,
and determining their ecological significance. In addition, organisms display a certain morphology as a
result of adapting to external forces; tending to evolve in order to achieve maximum level of fitness to
survive unfavorable conditions in its own habitat. Several of these morphologies are commonly used in
the field of microbiology, and some of which are the following: coccus for circular or ovoid in shape, rod
or bacillus for cylindrical, spirilla for spiral (Madigan et al. 2019).
Although morphology has served as an important tool in order to initially and generally determine
a specific microorganism, it rarely provides a concrete description for other properties of the cell. For
instance, there are several bacillus-shaped organisms that are classified in different domains, however,
still appear very similar; therefore, creating dubiety on where to primarily categorize each specimen upon
initial observation. However, in this study, morphologies of some specimens (mentioned in Materials and
Methods section) were only observed due to availability constraints.
Another common property that some cells possess is their capability of self-propulsion or motility.
Organisms use this in order to respond to changes in environmental conditions via transposition or
movement. In addition, this can be further subdivided into two types: Brownian movement, and True
motility.
By definition, brownian movement is the random motion of microorganisms, present in fluids,
that is often due to the constant collisions or interactions with other particles present in its surrounding
environment. On the other hand, true motility is the ability of the organism to move by itself via their
possessed external structures.
Organisms, such as bacteria, generally use the flagellar movement to exhibit true motility. This
motion is facilitated by the structure called flagella, a thread-like organelle that extends from the surface
of the cell with varied positioning or number. Another example of this kind is the cork-screw motion. This
is often responsible for the movement of the spirochetes, a type of bacteria. Consequently, this group
does not possess a flagella, rather, they have axial filaments to aid in the so-called cork-screw motion. In
addition, gliding movement is also considered to be a mechanism under true motility. This type is often
done through: a.) slime extrusion, b.) twitching, or c.) ratcheting, and is the sole movement cyanobacteria
and myxobacteria perform.
In relation to motility, protists also have their own way of relocating themselves in response to
external stimuli. These microorganisms also possess external structures that aid not only in movement,
but also for different survivability purposes (e.g. Paramecium use cilia for feeding). Given this, one of the
primary locomotory structures these organisms have are cilia. These are tiny hair-like organelles that
come in large numbers causing vibrations to produce currents, thus resulting to movement. Another
mechanism is, similar to most organisms, the use of the flagella. Lastly, pseudopods are extensions of the
cytoplasm to exhibit the movement of some organisms like amoeba. Given these, the experiment will
mainly focus on the motility of the following specimens: Escherichia coli, Staphylococcus aureus, and
microorganisms in a hay infusion. This property of the said samples will be observed under a compound
light microscope, however, will be initially prepared by the students through the wet mount technique
and hanging-drop technique.
The wet mount technique is one of the simplest ways in preparing live specimens. This technique
is usually done in order to view and observe the motility of living organisms. For this study, microbes in a
hay infusion, a nutrient media for growing ciliates, was utilized in order to execute the wet mount method.
On the other hand, the hanging-drop technique is a well-established method in the field of microbiology
in order to view unstained living microorganisms. This essentially employs a depression slide where a drop
of fluid consisting the specimen, in this case, E.coli and S. aureus, hangs from the cover slip.
In order to further understand the fundamentals of microbiology, the basic properties of a
microorganism must be observed via different preparation techniques. Hence, the study has the following
objectives:
● To differentiate the morphology of microorganisms
● To perform wet mount and hanging drop technique
● To differentiate true bacterial motility from Brownian movement

II. Materials and Methods

A. Morphological Structures of Microorganisms.
Observing Bacterial Morphology. Prepared slides of various specimens of the microbes
were obtained from the slide box provided for the class. For bacteria, five (5) samples
were utilized: Bacillus subtilis, Mycobacterium tuberculosis, Shigella dysenteriae,
Alcaligenes faecalis, and Corynebacterium diphtheriae. The specimens’ morphologies
were then observed under a compound light microscope using different objective⏤low
power objective (LPO), high power objective (HPO), and Oil Immersion objective (OIO).
Each were cleaned using lens paper with xylene before being utilized for the activity.

Utilization of Oil Immersion Objective. In using OIO, the specimen was initially focused
using the objectives with lower linear magnification. Upon switching from HPO to OIO, a
drop of immersion oil was placed on the cover slip of the prepared slide before carefully
swinging the objective into place.

Observation of Fungal, Algal, and Protozoan Morphology. The procedure for the
bacteria specimens was repeated for fungi, algae, and protozoa. For fungi, prepared slides
of the following were used: Aspergillus, Penicillium, Candida albicans, Saccharomyces
cerevisiae, and Rhizopus. On the other hand, for protozoa, Blastocystis hominis,
Chilomastix mesnili, Giardia lamblia (trophozoite), Paramecium, and Plasmodium
falciparum, were observed in order to identify each microorganisms’ specific morphology.
Lastly, for algae, only three specimens were examined in order to identify their distinct
morphologies. The following were the samples used for observing algal morphology:
Euglena sp., Zooplankton, and Green algae.

B. Motility of Microorganisms
Wet Mount Technique. The wet mount technique was utilized by the researchers in order
to prepare the slides of the hay infusion. A small drop of the prepared hay infusion was
placed on the glass slide. Afterwards, a coverslip was carefully placed. This is to ensure
that the number of air bubbles would be minimized. The HPO was then used to observe
the slide under the microscope. The motility of the protozoans were then observed.
 Hanging-drop Technique. The hanging-drop technique was utilized by the researchers to
prepare the slides of S. aureus and E. coli. On the cover slip, a small amount of petroleum
jelly was placed on all sides of the cover slip. This is to ensure that the bacteria would not
spill. The culture tube was then opened, and with the help of the dropper, transferred the
bacteria in it to the cover slip with petroleum jelly. On top of the bacteria, the depressed
side was placed on top of the bacteria, then inverted. This then made the bacteria hang
from the cover slip. The two bacteria were then observed by the researchers using the
OIO, and afterwards the motility of the organism were noted.

III. Results and Discussion

A. Morphological Structures of Microorganisms

BACTERIA MORPHOLOGY

Observing Specimens for Bacterial Morphology. Five (5) specimens of bacteria were utilized in
order to observe different morphological features present in the domain: a.) Corynebacterium
diphtheriae, b.) Spirillum lanatumBacillus subtilis, c.) Mycobacterium tuberculosis, d.) Shigella
dysenteriae, and e.) Alcaligenes faecalis. Most of the samples were viewed under a compound
microscope, using different objectives. However, the oil immersion objective was commonly used
for viewing most of the slides since it is generally known to have the highest magnification to view
organisms in a compound microscope. In addition, it was kept in mind to occasionally use lens
paper with xylene to clean the objectives.

Principle of Using Immersion in Microscopy. Utilizing immersion oil is major part when viewing
organisms under the OIO. In order to obtain a clearer image of what is being observed, a drop of
immersion oil must be placed on the cover slide, as the oil, guides the light through the object
being observed. This is caused by the equality of the refractive index of the immersion oil and
the glass slide, which has a space filled with air. In the space between an objective lens and a glass
slide which is filled with air, there is what we call a refractive index where light is lost or scattered.
However, the higher the magnification, as is the case with 1000x magnification, the more likely
for the light to be scattered. In order to achieve a clearer image in such a high magnification, one
must apply immersion oil on the slide, which directs the light through the objective despite the
present refractive index.

Principle of Xylene in Microcopy. In order to remove the immersion oil from the lenses and the
glass and cover slides, xylene must be used. The usage of xylene in removing the oil is highly
important as it is soluble to it, unlike water and alcohol. However, due to its toxicity, it must be
used with care.
Figure 1. Cell shape and arrangement of Corynebacterium diphtheriae (1000X)

Figure 2. Cell shape and arrangement of Spirillum lanatum (1000X)

Figure 3. Cell shape and arrangement of Mycobacterium tuberculosis (1000X)

 Figure 4. Cell shape and arrangement of Shigella dysenteriae (1000X)

Figure 5. Cell shape and arrangement of Alcaligenes faecalis (1000X)

Bacterial Characteristics. Bacteria are unicellular organisms thank lacks membrane bound
organelles and does not exhibit a true nucleus (Molnar and Gair 2013). In this regard, bacteria are
considered to be chemoorganotrophic since they harness energy from the oxidation instigated by
reduced organic compounds (Amils 2014). Bacteria can also be classified as gram positive or gram
Negative. Gram positive bacteria exhibit thick peptidoglycan layer (90%), presence of teichoic
acid, minimal presence of lipids, absence of outer membrane, and absence of periplasmic space.
Gram negative bacteria, on the other hand, exhibit thin peptidoglycan layer (10%), absence of
teichoic acid, presence of lipopolysaccharide surface layer, presence of outer membrane, and
presence of periplasmic space. Bacteria may also have varying morphologies, specifically in their
cell size, shape, and arrangements. Bacterial shape examples include, but are not limited to,
coccus, bacillus, spirillum, spirochetes, filamentous, and budding/appendage, while bacterial
arrangement examples include, but are not limited to, staphylo, diplo, strepto, sarcina, and tetrad
(Madigan et al. 2019). Correspondingly, some bacteria are characterized to be photosynthetic and
that some of them exhibit true bacterial motility, which may be in the form of flagellar motion,
corkscrew motion, or slime layer gliding motion (Kaiser 2018). Consequently, some bacteria also
exhibit assisted motility through the Brownian motion which is the erratic and random movement
of microorganisms instigated by collisions with molecules in a liquid or aqueous environment
(Germain et al. 2016).

Bacterial Specimen Descriptions. Figure 1 displays Corynebacterium diphtheriae that were

observed under oil immersion objective (1000x). Corynebacterium diphtheriae are Gram positive
and exhibit a club-shaped rod. They can have single, paired, V form, palisade, or clustered cellular
arrangements (Ryan 2004). These bacteria are the etiological agent of diphtheria, a disease that
infects and thickens the back of the throat (Centers for Disease Control and Prevention 2018).
Figure 2 displays Spirillum lanatum that were observed under oil immersion objective (1000X).
Spirillum lanatum is microbiologically described as a corkscrew like, gram-negative, and motile
helical bacteria with tufts of whiplike flagella at each end (Sheffer 2019). Figure 3 displays
Mycobacterium tuberculosis that were observed under oil immersion objective (1000x). These
bacteria are slightly curved or straight rods which can branch from 0.2 to 0.6 micrometer by 1.0
to 10 micrometer. They are acid-fast, aerobic, non-spore forming, non-motile, and slow growing
bacteria (Pfyffer 2007). Figure 4 displays Shigella dysenteriae that were observed under oil
immersion objective (1000x). These bacteria are Gram negative, rod-shaped and pathogenic. They
are non-motile, non-encapsulated, virulent (produces potent cytotoxin), and facultative
anaerobes that do not ferment or slowly ferment lactose (Schroeder and Hilbi 2008). Figure 5
displays Alcaligenes faecalis that were observed under oil immersion objective (1000x). These
bacteria are Gram-negative and appears rod-shaped and motile under a microscope. They are
positive by the oxidase test and catalase test, but negative by the nitrate reductase test. A.
faeucalis are alpha-hemolytic and requires oxygen; they can be grown at 37 °C, and forms colonies
that lack pigmentation (Rehfuss and Urban 2005).

FUNGAL MORPHOLOGY

Viewing Molds. Molds are fungi that grow in multicellular filaments called hyphae, which results
in discoloration and a fuzzy appearance. By looking at the hyphae, it is possible to identify the
kind of mold it is. Since these molds have a size range of 3-40 microns, it is necessary to use the
oil immersion objective or OIO in viewing mold specimen under the microscope to vividly see its
microscopic parts, such as the sporangium.
 Figure 6. Cell shape and arrangement of Aspergillus (400X)

Figure 7. Cell shape and arrangement of Penicillium (400X)

Figure 8. Cell shape and arrangement of Candida albicans (1000X)

 Figure 9. Cell shape and arrangement of Saccharomyces cerevisiae (1000X)

Figure 10. Cell shape and arrangement of Rhizopus (100X)

Fungal Characteristics. Fungi are eukaryotic organisms that exhibit true nucleus and membrane
organelles. These organisms are usually multicellular, with the exception of single-celled yeast. In
line with this, fungi are intracellularly and structurally made up of hyphae which are individual
feathery filaments and collectively they are known as the mycelium (McDougal 2018). Mycelium
is the vegetative structure of fungi, whereas, it extends the area in which a fungi can locate
nutrients. It is worth noting that fungi are stationary or non-motile organisms but their mycelium
grow outwards to locate water and nutrients such as nitrogen, carbon, potassium, and
phosphorus, which the mycelium mobilizes the fruiting body so it can prevail to grow, develop,
and manufacture biomass. Hence, all fungi are heterotrophic since they absorb nutrients from
their environment (Munks 2018).

Fungal Specimen Descriptions. As observed in Figure 6, Aspergillus is a green mold. Aspergillus

spp. are ubiquitous, structured with conidiophores, with their conidium-bearing structures found
at the terminal end vesicle, which may be uniseriate, covered in a single-layer of conidia-bearing
phialides, or biseriate, covered in a single-layer of metulae and and a single-layer of phialides
(Verweij and Brandt 2007). Aspergillus spp. are present worldwide, found in most environments,
and is responsible for causing aspergillosis, a respiratory sickness, but most cases primarily occur
in the immunocompromised (Ryan 2004). Figure 7 shows a Penicillium sp. specimen viewed under
400x magnification. Penicillium genus consist of filamentous fungi that have green or white
coloration, made up of a dense felt of conidiophores, with phialides produced in a verity, from
singly to groups, having a brush-like appearance (Visagie et al. 2014). Penicillium are ubiquitous,
found worldwide, inhabiting substrates like soil, decaying vegetation, and are airborne, capable
of producing mycotoxins (Pitt et al. 2000), which can cause penicilliosis in humans, though most
infections are found in immunosuppressed hosts (Deshpande and Koppikar 1999). Figure 8. shows
Candida albicans under 1000X magnification. Of family Candidaceae, C. albicans appears as
encapsulated and diploid, and it is a polymorphic fungus that can occur commonly as yeast or
pseudohyphal forms, depending on environmental conditions (Ryan 2004). As this specimen is a
subculture, the colonies appear smooth while its original colonies would appear wrinkled and
rough (Ryan 2004). C. albicans is an opportunistic pathogen that causes life-threatening diseases
in immunodeficient and immunocompetent individuals (Schell 2006). One such disease is
candidiasis, manifested by ragged white patches forming on mucosal surfaces like the tongue or
palate (Ryan 2004). Figure 9 shows Saccharomyces cerevisiae under 1000X magnification. S.
cerevisiae has a simple ellipsoidal shape (Saito et al. 2004) and colonises the mucosal surfaces of
normal individuals (AU 2016). Though overall it is harmless, S. cerevisiae is often involved in
mucosal infections such as vaginitis and bloodstream related infections (AU 2016). Figure 10
shows Rhizopus sp. specimen under 100X magnification. Rhizopus genus is characterized by its
structural components: stolons, pigmented rhizoids, sporangiospore, apophysate, columellate,
and sporangia (AU 2016). Its sporangiospores are globose to ovoid shaped, unicellular, and are
hyaline to brown in color (AU 2016).

PROTOZOAN MORPHOLOGY

Figure 11. Cell shape and arrangement of Blastocystis (1000X)

 Figure 12. Cell shape and arrangement of Chilomastix mesnili (1000X)

Figure 13. Cell shape and arrangement of Giardia lamblia trophozoite (100X)

Figure 14. Cell shape and arrangement of Paramecium (400X)

 Figure 15. Cell shape and arrangement of Plasmodium falciparum (400X)

Protozoan Characteristics. Protozoans are unicellular, eukaryotic, and cell wall lacking organisms
that belong in the Kingdom Protista. These organisms exhibit trophozoite which is utilized for
vegetation, reproduction, and feeding. As such, protozoans demonstrate phagocytosis. In this
regard, protozoans, being animal-like protists, are characterized as heterotrophs.
Correspondingly, these organisms exhibit at least one nucleus and they also constitute
intracellular structures. Additionally, some protozoans are capable of motility. Moreover,
protozoans may thrive in water environments, soil environments, and animals (Kaiser 2014).

Protozoan Specimen Descriptions. Figure 11 shows a Blastocystis specie observed under oil
immersion objective (1000X). Blastocystis is a genus of parasitic protozoans with Blastocystis
hominis being the most common one. They could cause an infection called blastocystosis to
humans which has symptoms like diarrhea and abdominal pain. The specimen appears to be
circular though Blastocystis spp. can appear in other morphological forms (Centers for Disease
Control and Prevention 2014). Figure 12 shows Chilomastix mesnili observed under oil immersion
objective (1000X). This protozoan is a non-pathogenic flagellate that exhibits a commensalistic
behaviour when ingested through the human gastrointestinal tract. Its cyst form resembles that
of a lemon shape and measures 6–9 µm in length while its trophozoite form is pear-shaped with
tapered posterior end and around 6–24 µm in length (Centers for Disease Control and Prevention
2019). Figure 13 shows Giardia lamblia trophozoite observed under low power objective (100X).
This protozoan flagellate can cause the disease giardiasis, an infection in the small intestines,
through the ingestion of its cysts. Its trophozoite form is pear-shaped and around 10–20 µm in
length (Centers for Disease Control and Prevention 2017). Figure 14 shows Paramecium observed
under high power objective (400X). This specimen appears to have a slipper shape with a blunt
posterior and a pointer anterior end. The size of Paramecium may range from 170-350 µm in
length (Vidyasagar, 2016). The nucleus and other intracellular parts were evident from the
specimen as well as the cilia that could be vaguely seen. Figure 15 shows Plasmodium falciparum
observed under high power objective (400X). These are blood parasites that can cause the disease
popularly known as malaria. Plasmodium falciparum and other species of Plasmodium are
transmitted to humans via mosquitoes thus infecting them as immediate hosts. The specimen
appears to be circular in shape and possibly in its schizont form (Centers for Disease Control and
Prevention 2017).

ALGAL MORPHOLOGY

Figure 16. Cell shape and arrangement of Euglena

(400X)

Figure 17. Cell shape and arrangement of Zooplankton (1000X)

Figure 18. Cell shape and arrangement of Assorted Green Algae (400X)

Algae Characteristics. Algae are plant-like protists that can be unicellular or multicellular. In line
with this, algae exhibits cell nucleus and membrane bound organelles. Most importantly, algae
 are corroborated to be photosynthetic, with the exception of Prothoteca sp., due to its
chlorophyllous attribution, thus making them autotrophic. Algae can also be colonial in nature.
Additionally, algae exhibit simple thallus and lack cell differentiation. Their ubiquitous nature
justify that they are found in various habitats such as fresh waters, marine waters, and even
extreme environments for some. Algae are diverse in structure and habitat, hence, there is also a
lot of diversity in their reproduction and perennation mechanisms (Sahoo and Baheja 2015).

Algae Specimen Descriptions. Being the most studied member of the class Euglenoidea, the
Euglena is a genus of 1,000 species of single-celled flagellated microorganisms which possess both
animal-like and plant-like characteristics (Bailey, 2019). Under the microscope, the Euglena
appears greenish in color with dark spots inside, extending its body with a tail-like structure or a
flagella. (Microscopemaster.com, n.d.). The description fits the specimen shown in Figure 16,
possessing the same characteristics the website had mentioned.
Along with phytoplankton, zooplankton are organisms that are carried by water currents
due to their miniature size, which make up the majority of the planktonic food supply. They also
play a significant role in the cycling of elements in the ocean. (Keister et al., 2012). Based on Figure
17, the specimen appears to be long and of a red-like color may be due to the stain, with what
appears to be alternating sections of colors or stripes.
As a member of the division Chlorophyta, the green algae comprises of 9,000 to 12,000
species. They also serve as the sources of food and oxygen of the different aquatic organisms,
mostly being freshwater species as it is where they are habituated. (Vidyasagar, 2016). Under the
microscope, it is described to possess a greenish color and of a wide variety of shapes.
(Microscopemaster.com, n.d.). Since Figure 18 exhibits more than one type of green algae,
therefore, different shapes can also be observed, as some are circular, some are cluttered
altogether, and some are long.

B. Motility of Microorganisms
 Figure 19. Wet Mount of Hay Infusion(400x)

Figure 20.Hanging Drop Slide of Escherichia coli (400X)

 Figure 21. Hanging drop slide of Staphylococcus aureus (400X)

Observation of Hay Infusion. In order to create a hay infusion set up, either a small amount of
hay, dried grass, or moss must be added to water. This set-up allows the microbes and other
microorganisms to survive as nutrients from the sugars available from the dried grass are obtained
by them. The bacteria then serves as food for the protozoans. Since microbes are usually small, it
is required that they be observed under the microscope using the wet mount technique.
Microorganisms, including protozoans such as amoebae, diatoms, may be observed through hay
infusion.

Bacteria, algae, and other protozoans are consumed by the protozoan, a single-celled eukaryote.
An example of a protozoan is an amoeba, which has no specific shape and move by utilizing
pseudopods. Another is the Paracelium which is a unicellular ciliate that thrives in marine
environments. On the other hand, cells with one or more whip-like tails or flagella are called
flagellates. A mayor algae group, which includes microalgae, are Diatoms. Waterways and oceans
are the usual environment these diatoms thrive in. Specifically, microalgae exists as a unicellular
species, and may be found individually or in clusters.

Protozoans are single-celled eukaryotes, who feed on bacteria, algae, and even other protozoans.
Amoeba serves as an example of protozoans, that have no specific shape, and move with the use
of pseudopods. Paramecium is a unicellular ciliate, which usually thrives in marine environments.
Flagellates are cells with one or more whip like tails called flagella, which are made out of flagellin.
Diatoms is a mayor algae group, specifically microalgae. These diatoms mostly thrive in waterways
and oceans. Microalgae exists as a unicellular species, and may be found individually or in clusters.
Difference between Brownian Movement and True Motility. Motility is associated with
chemotaxis or the ability to orientate along certain chemical gradients. Eukaryotic cells can move
through different locomotor organelles, such as cilia, flagella, or pseudopods. Prokaryotes move
through propeller-like flagella unique to bacteria or through special fibrils that instigate a gliding
form of motility. Majority of spiral bacteria and about half of the bacilli are motile, whereas, none
of the cocci are motile. In this regard, the capacity for true motility or self-propulsion is
demonstrated in an extensive variety of cells, but not all of them. As such The bacteria that show
no motion are denoted as non-motile. In lieu with this, in an aqueous environment, these same
bacteria appear to be moving erratically. Such erratic movement is caused by Brownian
movement which occurs from the random motion of the water molecules bombarding the
bacteria and instigating them to move. Hence, majority of motile bacteria move through the use
of flagella or the thread like locomotor appendages protruding outward from the plasma
membrane and cell wall. Motile cells may exhibit one flagellum, but most have several. Each
flagellum has a very rigid, helical structure and actual motility occurs from the rotation of the
flagellum. Motility by means of flagella is particularly important for the identification of
microorganisms (Virtual Amrita Laboratories 2011).

Nonetheless, in reiteration, some bacteria exhibit true bacterial motility, which may be in the
form of flagellar motion, corkscrew motion, or slime layer gliding motion (Kaiser 2018).
Furthermore, some bacteria may also exhibit assisted motility through the Brownian motion
which is the erratic and random movement of microorganisms instigated by collisions with
molecules in a liquid or aqueous environment (Germain et al. 2016).

Observation of Motility of Bacterial Cultures. As discussed, there are two main categories for
describing the motility of an organism, either Brownian movement or True motility. Given the
specimens used for the experiment, it was found that Escherichia coli (see Figure 20) was the
microorganism that exhibited true motility. Given this, they are able to present this kind of
mechanism because they possess thread-like external structures called flagella (Mittal et al.
2003). On the other hand, Staphylococcus aureus is an example of an organism that displays
Brownian movement. Although they exhibit motion when observed, it is potentially due to the
collision of it to various particles to its surrounding environment (e.g. water particles) (Pollitt and
Diggle 2017).

Other Microorganisms. Microorganisms are deemed to be the most abundant and most primitive
life forms on Earth. This study has mainly focused on bacteria, fungi, algae, and protozoa.
However, it is known that there are other classifications or groups or domains present in various
habitats and ecosystem. Some of which are archaea, helminths, virus, and subviral agents. Given
these, it is important to understand and know some of their characteristics.

Archaea Characteristics. Unlike bacteria, archaeans, along with the eukaryotes, lack
peptidoglycan layer in their cell walls, whereas, they exhibit a pseudopeptidoglycan or
pseudomurein that constitutes N-acetyltalosaminuronic acid and N-acetylglucosamine binded by
 the β-1,3-glycosidic bonds, and L-form amino acids. Such chemical complex of the pseudomurein
or pseudopeptidoglycan resist lysozyme. Similarly, the archaeal membrane exhibit ether bonds
that link fatty acids and glycerol and repeating isoprene units while it does not exhibit true fatty
acids. Additionally, its lipid monolayers are exhibited in a 1 long chain; its diglycerol tetraethers
justify that the lipid monolayers are more resistant to heat, i.e. hyperthermophiles, while its
chermarcheol is demonstrated by the methanogens and halophiles (Madigan et al. 2019). As far
as methanogens are concerned, methanogenesis, which is the production of methane as a
metabolic by-product, occurs exclusively in domain archaea, particularly Euryarcheota
(Niederberger 2019).

Helminth Characteristics. Helminths, or parasitic worms, are eukaryotic parasites known for their
capability to feed and live on living hosts. Similarly, it can instigate the disruption of the hosts’
nutrient absorption by utilizing all nutrients that mobilizes through the intestinal tract. In this
regard, helminths are characterized by the presence of attachment organs which include dentary
plates, teeth, lips, hooks, and suckers (Lumen Boundless Microbiology 2012).

Virus Characteristics. Viruses are acellular and they exhibit living traits and replication capabilities
once it has infected a host. Henceforth, viruses depend on a host cell for energy, metabolic
intermediates, and protein synthesis. It may also possess own genetic information and
extracellular structures. These viruses may also act as bacteriophages wherein they can attach to
a bacterium surface and infect it. Viruses can be sub-classified into viroids and prions, the former
containing nucleic acids and not exhibiting protein coat while the latter not containing nucleic
acids and exhibiting protein coat. Correspondingly, viroids cause plant diseases while prions cause
mad cow disease. In further regard, viruses refer to the infected host cell while virions refer to
the vector or virus particle itself and its inability to generate energy and replicate (Madigan et al.
2019).

Subviral Agents Characteristics. Subviral agents are comprised of three kinds which are satellite
viruses, viroids, and prions. These transmissible agents are subviral since they are less than a virus
in some attributions. Firstly, satellite viruses depends on another virus, a host, or helper virus for
propagation. This type of subviral agent is morphologically indistinguishable from regular viruses
but it is worth noting that a satellite virus is often denoted as “a parasite of a parasite” since it
depends on another parasite or a virus. Secondly, viroids are subviral agents found in plants. These
are comprised of RNA only and lack protein component. It is a circular RNA molecule itself
approximately only 0.3 kb in length. Nevertheless, the viroid RNA is transmissible and instigates
pathogenic lesion in plants. Thirdly, prions are transmissible agents that are comprised of protein
only and lack nucleic acid components. These are also etiological agents of neurodegenerative
diseases (Ryu 2017).

IV. Conclusion and Recommendations

Eighteen prepared specimen slides that include bacteria, fungi, protozoans, and algae, a single
wet mount of a hay infusion, and two hanging-drop slides of S. Aureus and E. Coli were observed by the
researchers. For the bacterial specimens, each of their morphologies such as shape and arrangement were
determined as well as the Gram reaction of each bacterium. Their biology and the possible diseases and
conditions that they can cause were also discussed. For the fungal specimens, each of their morphologies
mainly appearance were explored. Their living conditions and infections that they could cause to humans
were also discussed. For the protozoan specimens, each of their morphologies like their cyst and
trophozoite stages were described along with possible illnesses that they could deliver to humans. For the
algal specimens, each of their morphologies including their animal-like and plant-like characteristics and
their environment niche were detailed.
The preparation of the hay infusion wet mount was done properly by the researchers. Microbes
were observed from the specimen such as bacteria, algae, and protozoans particularly a live Paramecium
organism. Slide specimens of S. aureus and E. coli were prepared by the researchers through the hanging-
drop technique. However, after attempting to find these bacteria under the microscope, the researchers
were unsuccessful in locating them. This may be the cause of a contaminated dropper. Instead of using
new and clean droppers in placing the bacteria on the cover slide, only one dropper dipped in 70% ethanol
was used by all of the groups. When the single dropper was used by the researchers, it was doused with
ethanol; thus resulting to some bacteria being killed. This led the motility of the bacteria in the hanging-
drop slide to be unobserved by the researchers. In the end, the motility of both bacteria were determined
with the E. coli exhibiting true motility while S. aureus moves through Brownian movement.
The different microorganisms took a considerable amount of time to be located by the
researchers due to the microscopes’ corresponding issues such as an unclear high power objective (HPO),
and a malfunctioning oil immersion objective (OIO). Therefore, it is recommended that the high power
and oil immersion objective lenses of some microscopes in the laboratory be replaced with clear and
functioning lenses to be able to efficiently and properly view microbial specimens.
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