MICROALGAE SPECIES IDENTIFICATION BY THEIR WAY

OF LIFE

By:
Name : Rahmi Mutia Mawardi
Student ID : B1B015041
Group :2
Entourage :I
Assistant : Lukman Adi Nugroho

PRACTICAL REPORT OF PHYCOLOGY

MINISTRY OF RESEARCH, TECHNOLOGY, AND HIGHER EDUCATION

JENDERAL SOEDIRMAN UNIVERSITY
FACULTY OF BIOLOGY
PURWOKERTO
2017
 I. INTRODUCTION

A. Background

Microalgae is a microbial plant that plays an important role in the

environment as a primary producer, in addition to bacteria and fungia around us.
Most microalgae are photosynthetic, have chlorophyll to capture solar energy and
carbon dioxide into organic carbon that is useful as a source of energy for consumer
life such as copepods, molluscs larvae, shrimp and others. In addition to its role as a
primary producer, the byproduct of the microalgae photosynthesis of oxygen also
contributes to the respiration of surrounding biota. Knowledge of physiology has
grown rapidly after various types of algae with their respective characteristics
successfully cultured. Various institutions in the world have kept a collection of
microalgae cultures that can potentially be utilized for various applications
(Panggabean, 2007).
Microalgae are vegetable microorganisms that live hovering in water,
relatively have no motion so that its existence is influenced by the movement of
water and able to photosynthesize. Microalgae are generally single-celled or yarn-
shaped, as plants and are known as phytoplanktons. Phytoplankton have a green leaf
substance (chlorophyll) that plays a role in photosynthesis to produce organic matter
and oxygen in water. As the basis of the chain in the food cycle in the sea,
phytoplankton become a natural food for zooplankton both small and mature. It can
also be used as an indicator of the fertility of a water. However, certain
phytoplankton have a role to degrade the quality of marine waters if the amount is
excessive. The example Dinoflgellata class of the body has a chromatophore that
produces toxin (poison), in a state of blooming can kill fish (Prasetyo, 1967).
Microalgae are a group of small-sized plants that belong to the algae class.
The diameter is between 3-30 nanometers, either single cells or colonies that live in
all freshwater and marine areas commonly called phytoplankton. In the microbial
world, microalgae include eukaryotic, generally photosynthetic with green
photosynthetic pigments (chlorophyll), chocolate (phycoxantin), and red
(phycoerythrin) (Graham, 2000). The microalgae morphology is unicellular or
multicellular but there is no clear division of tasks on the component cells. That's
what distinguishes microalgae from high-level plants.
 B. Purpose

The purpose of this experiment was to knowing the diversity of microalgae in

terms of different way of live in the wildlife.

C. Literature

According to that identification is the task of searching for and recognizing

the traits of individual taxonomy and incorporating it into a taxon. Identification
procedure based on deductive thinking. The definition of identification is very
different from the notion of classification. Often these two meanings are mixed up,
whereas the classification procedure is inductive. Identification relates to the small
number of taxonomic traits (ideally one feature), will carry the specimen into a
sequence of identification keys, while the classification relates to evaluating a large
number of traits (Labeda, 1990).
Microalgae are vegetable microorganisms that live hovering in water,
relatively have no motion so that its existence is influenced by the movement of
water and capable of photosynthesis. Microalgae are generally single-celled or yarn-
shaped, as plants and are known as phytoplanktons. Phytoplankton have a green leaf
substance (chlorophyll) that plays a role in photosynthesis to produce organic matter
and oxygen in water. As the basis of the chain in the food cycle in the sea,
phytoplankton become a natural food for zooplankton both small and mature. It can
also be used as an indicator of the fertility of a water. However, certain
phytoplankton have a role to degrade the quality of marine waters if the amount is
excessive. The example Dinoflgellata class of the body has a chromatophore that
produces toxin (poison), in a state of blooming can kill fish. Algae are primary
producers in aquatic ecosystems and are unicellular organisms, filaments and
asexually proliferating. The way of life can be attached or drift as phytoplankton.
Algae by size can be divided into microalgae and macroalgae. Microalgae are algae
that are so small that they can not be seen by the naked eye. Microalgae are also
scattered in marine waters. Plant-like protists are known as Algae. Microalgae are a
group of small-sized plants that have thallus and chlorophyll with habitat spread
throughout the freshwater, brackish, marine and terrestrial waters. Microalgae
contain chlorophyll which can alter inorganic compounds into organic compounds
using light energy through the process of photosynthesis to meet the nutritional needs
(Jati, 2007).
 Identification of plankton, identification is done by using binocular
microscope with 400 times magnification. As much as 0.03 ml of the homogenized
sample, dripped on the surface of a concave glass object, then covered with a glass
cover, and observed by shifting the glass object horizontally and vertically, so that all
glass cover surfaces are observed. Each observed plankton was drawn, then
morphologic identification at the genus level using plankton identification book
Illustration of The Marine Plankton of Japan (Yamaji, 1976) and Planktonology
(Sachlan, 1972). Changes in diatom abundance are also observed (Astuti et al.,
2012).
The three most important classes of micro-algae in terms of abundance are
the diatoms (Bacillariophyceae), the green algae (Chlorophyceae), and the golden
algae (Chrysophyceae). All these micro-algae are eukaryotes distinguished by the
presence of a nucleus and separate organelles for photosynthesis (chloroplasts) and
respiration (mitochondria). In this report, cyanobacteria (Cyanophyceae or blue-
green algae) are also referred to as micro-algae, e.g. the species Spirulina
(Arthrospira platensis and A. maxima). The cyanobacteria are part of the eubacteria
and are prokaryotes lacking a membrane-bounded nucleus. There are thousands of
different species of micro-algae and cyanobacteria (Sander & Ganti, 2010).
 II. MATERIAL AND METHODS

A. Material

The materials that used in this practice are microalgae sample from water,
soil, lichen (simbionts), and sterile aquadest.
The tools that used in this practice are microscope, object glass, cover glass,
pipette drop, microtom/razor blade, petridish, and pinset.

B. Methods

1. Microalgae samples from water taken with plankton net the put into the bottle.
Samples of from soil diluted with sterile distilled water then put into the bottle.
For observation, microalgae sample in the bottle is homogenized first.
2. Using a pipette microalgae samples is taken then dripped on object glass and
covered with a glass cover subsequently observed under a microscope.
3. Smaples of symbionts in a way lichen across incision is made and then observed
under a microscope.
4. Identifying and classifying types of microalgae are achieved using following
identification book.
 III. RESULT AND DISCUSSION

A. Result

Table 3.1 Results of Microalgae Identification Entourage I

No. Group Microalgae

1. 1 Coelastrum microporum, Crucigenia sp.
2. 2 Pediastrum gracillimum, Haematococcus sp., Chlorella
pyrenoidosa
3. 3 Chlorella sp.
4. 4 Chlorella sp.
5. 5 Pediastrum duplex var. subgranulatum
6. 6 Not found microalgae

Figure 3.1 Pediastrum gracillimum Figure 3. 2 Haematococcus sp.

Figure 3.3 Chlorella pyrenoidosa

 B. Discussion

The results of microalgae identification obtained by group 2 of entourage I

were Pediastrum gracillimum, Haematococcus sp., Chlorella pyrenoidosa. Other
groups get different species. Some get microalgae with the same genus but different
species. This can happen because of the very abundant microalgae diversity in pond
water. In same case can also find the same microalgae species but differ in
identification process.
The classification of Pediastrum gracillimum according to Wilcox & Floyd
(1988) there are:
Kingdom : Plantae
Phylum : Chlorophyta
Class : Chlorophyceae
Order : Sphaeropleales
Family : Hydrodictyaceae
Genus : Pediastrum
Species : Pediastrum gracillimum
Pediastrum gracillimum is a type species of fresh water green algae in the
genus pediastrum. They form nonmotile coenobia (colonies) with a fixed number of
cells. These coenobia are flat and have a circular shape. The cell bodies are
polygonal, are granulated and have horn-like projections. They reproduce these
autocolonies asexually. The colonies usually contain 8 to 32 cells, with examples of
4,64 or 128 occurring rarely (McManus & Lewis, 2005).
The classification of Haematococcus sp. according to Lorentz & Cysewski
(2000) there are:
Kingdom : Plantae
Phylum : Chlorophyta
Class : Chlorophyceae
Order : Chlamydomonadales
Family : Haematococcaceae
Genus : Haematococcus
Species : Haematococcus sp.
Haematococcus sp. is a freshwater species of Chlorophyta from the family
Haematococcaceae. This species is well known for its high content of the strong
antioxidant astaxanthin, which is important in aquaculture, and cosmetics. The high
amount of astaxanthin is present in the resting cells, which are produced and rapidly
accumulated when the environmental conditions become unfavorable for normal cell
growth. Examples of such conditions include bright light, high salinity, and low
availability of nutrients (Dore & Cysewski, 2003).
The classification of Chlorella pyrenoidosa according to Merchant & Andre
(2001) there are:
Kingdom : Plantae
Phylum : Chlorophyta
Class : Trebouxiophyceae
Order : Chlorellales
Family : Chlorellaceae
Genus : Chlorella
Species : Chlorella pyrenoidosa
Chlorella is a green algae that grows in fresh water, and Chlorella
pyrenoidosa is a species of this algae. Algae supplements might contain a
combination of green algae, blue-green algae and spirulina, so if you want this
particular species, buy a supplement labeled Chlorella pyrenoidosa from a reputable
manufacturer. Consult with a qualified health care provider before taking any herbal
supplements (Nakano et al., 2007).
Microalgae is a microbial plant that plays an important role in the
environment as a primary producer, in addition to bacteria and fungia are all around
us. Most microalgae are photosynthetic, have chlorophyll to capture solar energy and
carbon dioxide into organic carbon that is useful as a source of energy for consumer
life such as copepods, molluscs larvae, shrimp and others. In addition to its role as a
primary producer, the byproduct of the microalgae photosynthesis of oxygen also
contributes to the respiration of the surrounding biota. Knowledge of physiology has
grown rapidly after various types of algae with their respective characteristics
successfully cultured. Various institutions in the world have kept a collection of
microalgae cultures that can potentially be utilized for various applications
(Panggabean, 2007).
Microalgae is one of the organisms that can be considered ideal and potential
for serve as raw material for biofuel production. The content of lipid in biomass of
dry microalgae of certain species can reach above 50% with growth very fast. The
microalgae breeding process only takes 10 days to be ready Harvested so that the
mathematical productivity reached (120,000 kg biodiesel/Ha year). More than 20
times the productivity of palm oil (5,800 kg biodiesel/Ha year) and 80 times Fold
compared to castor oil (1,500 kg/biodiesel/Ha year). Microalgae have the potential to
produce very high amounts of biofuel biodiesel and bioethanol are potentially
renewable fuels caught the world's attention. Biodiesel and bioethanol produced by
agricultural crops using existing methods and its existence can not replace that fossil
oil fueled. The high potential of the material from microalgae that microalgae oil
contains lipids suitable for esterification or transesterification (Maharsyah et al.,
2013).
According to Isnansetyo & Kurniastuty (1995), the way of life microalgae
can be classified into:
1. Phytoplankton
Free living floating or floating in the water. How to move freely with the
flow of water (passive). There is an active called neuston.
2. Fitobentos
Life is attached to the substrate or something at the bottom of the waters.
Based on the size distinguished into bentos macroalgae and bentos microalgae.
Depending on the type of substrate, grass or aquatic plants and water currents.
Substrate type: stable eg rock and unstable eg sand.
3. Symbiotic algae
Live together and associate with other organisms. The advantage of the
symbionts is that the host gets food while the alga gets the protection/permanent
environment and the food substances. Losses of nutrient absorption/light for the
host is reduced or narrow.
4. Aerial algae
There are 2 types, growing in moist and sunny soil surface for
photosynthesis, Example: green algae in acid soil, Cyanobacteria in neutral soil;
Growing in the stone surface, between the rock and the many (endolitic), coccoid
form.
Some variations of cell shape can be found in unicellular algae may be
elongate or square-shaped elongated ball in addition to some unicellular having arms
or spines that are an extension of the cell wall. Many microalgae form cell filaments
that connect to each other. Microalgae form cell colonies that have a special pattern
and are determined by cell numbers cultural conditions will determine the
morphology of an organism and its variations (Borowitzka & Borowitzka, 1988).
The newest research about microalgae who researched by Rodrguez-Morales
et al. (2017), that are a comparison of the bioavailability of zinc (II) using metal
internalization by two microalgae species, Scenedesmus acutus and
Pseudokirchneriella subcapitata, and a hollow fiber supported liquid membrane
(HF-SLM) device as a chemical surrogated is performed. As metal speciation is
greatly influenced by different environmental variables, and metal ion bioavailability
is expected to be dependent on environmental variables, a multivariate approach is
used to account for multiple effects and the interactions on zinc internalization
behavior. To the best of our knowledge, this study is the first comparison of a
chemical system and two biological references that measures zinc (II) bioavailability
in complex aqueous media employing a multivariate approach.
IV. CONCLUSION AND RECOMMENDATION

A. Conclussion

Based on the result can be concluded that:

1. Microalgae that we obtained are Pediastrum gracillimum, Haematococus sp.,
Chlorella pyrenoidosa.

B. Recommendation

Suggestions for this lab is to practice more thoroughly in terms of the type of
result microalgae by using the software in order to avoid mistakes in determining the
type of gill microalgae species..
 REFERENCES

Astuti, R.P., P.T. Imanto, & Gede S.S. 2012. Kelimpahan Beberapa Jenis Mikroalga
Diatom di Perairan Pulau Gumilamo-Magaliho, Halmahera Utara. Jurnal Ilmu
dan Teknologi Kelautan Tropis. 4(1), pp. 97-106.
Borowitzka, M.A. & Borowitzka, L.J. 1988. Micro-algae Biotechnology. New York:
Cambridge University press.
Dore, J.R. & Cysewski, G.R. 2003. Haematococcus algae meal as a source of
natural astaxanthin for aquaculture feeds. Hawaii: Cyanotech Corporation.
Graham, L.W. 2000. Algae. New Jersey: Prentice Hall, Inc.
Isnansetyo, A. & Kurniastuty. 1995. Teknik Kultur Fitoplankton dan Zooplankton,
Pakan alami Untuk Pembenihan Organisme Laut. Yogyakarta: Penerbit
Kanisius.
Jati, W. 2007. Aktif Biologi. Jakarta: Ganeca Exact.
Labeda, D.P. 1990. Isolation of Biotechnological Organism from Nature. New York:
McGraw-Hill Publishing Company.
Lorentz, R. T., & G. R. Cysewski. 2000. Commercial potential for Haematococcus
microalgae as a natural source of astaxanthin. Trends Biotechnology, 18,
pp.160-167.

Maharsyah, T., Mustofa, L. & Wahyunoto, A.G. 2013. Efektivitas Penambahan Plant
Growth Promoting Bacteria (Azospirillum sp.) dalam Meningkatkan
Pertumbuhan Mikroalga (Chlorella sp.) pada Media Limbah Cair Tahu
Setelah Proses Anaerob. Jurnal Keteknikan Pertanian Tropis dan Biosistem,
3, pp.258-264.

McManus, H.A. & Lewis, L.A. 2005. Molecular phylogenetics, morphological

variation and colony-form evolution in the family Hydrodictyaceae
(Sphaeropleales, Chlorophyta). Phycologia, 44, pp.582-595.
Merchant, R.E. & Andre, C.A. 2001. A review of recent clinical trials of the
nutritional supplement Chlorella pyrenoidosa in the treatment of
fibromyalgia, hypertension, and ulcerative colitis. Alternative therapies in
health and medicine, 7(3), 7991.
Nakano, S., Takekoshi, H. & Nakano, M. 2007. Chlorella (Chlorella pyrenoidosa)
supplementation decreases dioxin and increases immunoglobulin a
concentrations in breast milk. Journal of Medicinal Food, 10(1), pp.13442.
Panggabean, L. M. G. 2007. Koleksi Kultur Mikroalgae. Oseana, 23(2), pp.11-20.
Prasetyo, I. 1967. Mikroalga. Jakarta: Erlangga.
Rodrguez-Morales, E.A., Miguel, E.R.S. & Gyves, J. 2017. Evaluation of a hollow
fiber supported liquid membrane device as a chemical surrogate for the
measurements of zinc (II) bioavailability using two microalgae strains as
biological references. Chemosphere, 171, pp.435-445.
Sachlan, M. 1972. Planktonologi. Jakarta: Direktorat Jendral Perikanan.
Sander, K. & Ganti, S. M. 2010. Life Cycle Analysis of Algae. Journal
Microbiology, 2(15), pp.704-714.
Wilcox, L.W. & Floyd, G.L. 1988. Ultrastructure of the gamete of Pediastrum
duplex (Chlorophyceae). Journal of Phycology, 24, pp.140-146.
Yamaji, I. 1976. Illustrations of the Marine Plankton of Japan. Japan: Hoikusha
Publishing Co., LTD.