TILAPIA (OREOCHROMIS NILOTICUS) MUCUS AS DESSICATING AGENT TO

ABSORB MOISTURE

A study presented to the

Faculty of Lemery Senior High School

Lemery, Batangas

In Partial Fulfillment

of the requirements for Practical Research 2

Grade 12 Senior High School

By:

Deomampo, Jan Dane Riyell F.

Maneja, Micaella Mae B.

Palicpic, Karen E.

Samson, Ysabellyreiss S.
 CHAPTER I

THE PROBLEM AND ITS BACKGROUND

This chapter presents the problem and the background of the study to give the initial idea
about this research study.

1.1. Background of the Study

Many people run across the issue of moisture. It has been proven that the presence in the
atmosphere of moisture or water vapor can alter the material's composition. It can cause molds,
rust, and bacteria that result to damage to the material. Due to this fact, every material needs a
specific type of desiccant.

Desiccants are drying agents that absorb moisture from materials to prevent it from being
harmed. A large amount of their moisture weight will be preserved by these desiccants and the
majority of the moisture inside the package will be adsorbed. Owing to their chemical properties,
they are either soluble or insoluble substances that adsorb water. Dry desiccants can absorb
moisture from air either by physical adsorption or by chemical reaction, and thus reduce the
humidity in the headspace of sealed containers (Chen, 2017).

Desiccants come in various forms and have found widespread use in food,
pharmaceuticals, packing, electronics and many manufacturing industries (Daou & Wang, 2005).
They can be incorporated into sachets, canisters, cards, or even into bottle closures. The overall
moisture sorption behavior of the packaged material (drug product and desiccant) can be
calculated from the weighted sum of the moisture sorption isotherms of the active
pharmaceutical ingredient (API), excipients and desiccant (Pellett, 2018).

The researchers came up with the idea of using fish skin mucus to develop a desiccating
agent to absorb moisture.

In a fish, a slimy, slippery substance called mucus coats the epidermal surfaces. A stable
physical or chemical barrier against invading pathogens is provided by fish skin mucus. It can
also be defined as a viscous colloid containing mucin-like antibacterial enzymes, proteins, water,
and more. The mucus has a wide range of functions including disease resistance, protection, as
well as respiration, ionic and osmotic regulation, reproduction, excretion, communication,
feeding and nest building (Ingram, 1980 ). The fish mucus sample can be obtained by gently
scraping the external fish surfaces avoiding ventral areas in order to avoid intestinal or sperm
contaminations.
 This study was conducted to find out the effectiveness of tilapia (oreochromis niloticus)
mucus as a desiccating agent to absorb moisture. The absorption rate will be tested out and be
compared to the commercially existing ones like silica gel, molecular sieves, and clay mineral.

1.2. Statement of the Problem

This study entitled “ Tilapia (oreochromis niloticus) Mucus as Dessicating Agent to

Absorb Moisture”was conducted to analyze the potential of fish mucus as a desiccating agent to
absorb moisture.

Specifically this study will seek answers to the following:

1. What are the properties of fish mucus in terms of viscosity?

2. What is the absorption rate of fish mucus?

3. Is there a significant difference between the absorption rate of fish mucus and the
commercially available ones?

4. How much moisture can the fish mucus hold?

5. Is there a significant difference between the desiccating efficiency of the fish mucus and the
existing ones in the market?

1.3. Scope, Limitation and Delimitation

This study focused on the efficiency of Tilapia mucus from the areas of Subic, Agoncillo
as dessicating agent to absorb moisture.

The material used in this study was acquired from the mucus of Tilapia fish by using
spatula to collect them. Samples of Tilapia mucus were collected from Tilapia fish by gently
scraping with a sterile spatula from the dorsal surface of the body; the ventral side of the body
was not considered in order to avoid intestinal and sperm contamination and mucus was
collected at regular intervals (10 attempts in 1 day). The mucus samples were frozen in ice at 0
⁰C to avoid bacterial growth to absorb the moisture well. Materials were used to investigate the
effectiveness of tilapia mucus. Set A will be tested using the alternative dessicating agent which
is mostly made up of tilapia mucus while set B will be tested using a commercialized dessicating
agent.

This study does not cover any other aspects other than the effects of Tilapia mucus as
dessicating agent to absorb moisture. In addition, any type of fish in Subic Ilaya, Agoncillo
except Tilapia are not included.
1.4. Significance of the Study

This study was conducted to determine the effectiveness of Tilapia (oreochromis

niloticus)Mucus as dessicating agent to Absorb Moisture.

Specially, this study may be beneficial to the following:

Community;

This study will be beneficial to the community for it will help have the opportunity to
make their own desiccants out of fish mucus which can be pretty available to anyone.

Businessmen;

Creating desiccating agent out of fish mucus have a potential of helping manufacturing
comoanies to make products at a very low cost. Thus, providing a more convinient source of
materials to use in their products.

Scientists;

This study will be helpful for scientists because the information gained out of this study
would introduce new and deeper knowledge comparwd to what they are already aware of about
fish mucus. This would help them discover other uses of fish mucus in the field of science.

Future researchers;

This study will provide knowledge and new information for future researchers that can
help them get ideas for creating their own research. It can also serve as one of their references
and related study for their own research study.

1.5. Hypotheses

H o 1There is no significant difference in the absorption rate between tilapia mucus and the
commercially one.

H o 2There is no significant difference in the dessicating efficiency between the tilapia mucus and
the commercially one.
 CHAPTER II

REVIEW OF RELATED LITERATURE AND STUDIES

This chapter presents the review of related literature and studies as well as the synthesis
to thoroughly discuss the related concepts in this research study to make the readers better
understand the context of this study.

2.1 Related Literature

Dessicants

Desiccants are drying agents that extract water from a wide range of materials. They are
either soluble or insoluble substances that adsorb water due to their chemical properties.
Examples include silica gel, bauxite, calcium sulfate and montmorillonite clay. Soluble agents
include calcium chloride and glycerol. Molecular sieves, alumina and activated charcoal also
make exceptional desiccants.

Desiccants are used to prevent humidity from damaging the contents of sealed packages.
This is the reason that so many products include small packages of silica gel and similar
materials. These desiccants can hold a large percentage of their weight in moisture, and will
adsorb the great majority of humidity within the package. Desiccants also find use in industrial
applications, such as humidity detection, waste removal and cleaning, and regenerative drying.

Regenerative dryers send air to a molecular sieve (or other) desiccant bed, where the
moisture is adsorbed. Later, this moisture is purged by sending heated air through the system;
this lowers the moisture retention capabilities of the desiccant and forces it to release stored
moisture into the heated air. Eventually, this moisture laden air is expelled and the regenerative
dryer cycle is complete.

Desiccants come in a wide range of shapes, from perforated canisters containing silica
gel to silica gel beads encased in specialized bags. Various substances, such as paper, cotton and
Tyvek, are used as desiccant packaging material. Typically commercial desiccants of this nature
will change color as they become full with water, thereby alerting the user that a new desiccant is
required.

(https://www.thomasnet.com/articles/chemicals/about-desiccants/)

Drying agents (also called desiccants) come in various forms and have found widespread
use in the foods, pharmaceuticals, packing, electronics and many manufacturing industries. A
desiccant is a hygroscopic substance that induces or sustains a state of dryness in its vicinity.
Ideally, it is chemically stable and chemically inert (i.e., silica). Unfortunately, this is not always
the case in the chemistry lab because the drying agent comes into direct contact with the solvent
and the chemical.

Many organic solvents are immiscible with aqueous solutions, but they are able to
dissolve significant amounts of water because of their polarity i.e., diethyl ether dissolves 7 % of
its weight in water while tetrahydrofuran is completely miscible with water. Unfortunately, water
is a compound that is very difficult to remove from many compounds, because they are either
holding on to it well (i.e., alcohols) or the compound itself is steam volatile. Bottom-line is that
the more polar the solvent is, the more hygroscopic it will usually be because it dissolves the
water better. Thus, removing water and other impurities from a solution can become an arduous
task but is necessary if the reagents are also sensitive towards water i.e., Grignard reagents or in
cases where water has a detrimental effect on the yield or rate of the reaction. In those cases,
drying agents like calcium hydride (CaH2), sodium metal (in combination with benzophenone)
or lithium aluminum hydride (LiAlH4) are used to chemically destroy the water in the solvent.
Those compounds are relatively reactive and difficult to handle and usually not used in lower
division undergraduate laboratories.

(https://www.chem.ucla.edu/~bacher/Specialtopics/Drying%20Agents.html)

The ability of a desiccant to attract water vapour depends on the difference in vapour
pressure between its surface and the process air. All desiccants attract moisture until they touch
equilibrium point with the surrounding air. The absorbed water vapour is usually removed from
the desiccant by regeneration in which desiccant is exposed to a regeneration airstream having
temperatures between 50 and 260°C. Generally, the capacity of solid desiccant is less than the
capacity of liquid desiccant but the use of composite desiccant materials may improve the
moisture adsorption capacity. In case solid desiccant for air dehumidification with low grade
waste heat as driving energy silica gel always performs better than other solid desiccant. Where
as in case of liquid desiccant the most suitable is LiCl because of its lower vapour pressure but
the major problem with LiCl is its high cost, this issue can be solve by combining it with
cheapest desiccant CaCl2. At the same time the composite desiccant of silica gel and LiCl has
adsorption capacity 2-3 times higher than that of pure silica gel. The limitation faced by
desiccant system is availability of regenerating heat to regenerate desiccant material. But the use
of solar energy and waste heat for regeneration of desiccant material will make the system more
economical. The use of desiccant system can solve lot of environmental problems well, as it can
also minimise the high demand of electrical energy for conventional air-conditioning system and
poor indoor air quality (R P Singh, V K Mishra, R K Das).Water and MucusMucus secretions
come from mucus-producing cells in the foregut, intestine, liver, and pancreas. Mucus contains
water and mucins. Mucins are highly glycated proteins with great water-holding capacity and are
produced by specialized epithelial cells located in the mucosal lining all along the alimentary
tract. Fish appear to lack salivary glands and instead have salivary cells dispersed among the
epithelial cells. The number of mucin-secreting cells varies between the esophageal, gastric, and
intestinal sections and so does the flow of mucus, from low in the mouth to high in the intestinal
regions. The quality of gut mucosubstances can vary with factors such as feeding habits, diet
composition, fish species, and developmental stage. The mucus of the gastrointestinal (GI) tract
may also contain immunoglobulins and antimicrobial components such as enzymes and peptides,
as well as organic compounds with antioxidative characteristics. Dead cells sloughed off from
the surface also add proteins and enzymes to the mucus. The secretion of mucus may be helpful
in removing harmful bacterial products by a washout effect. Mucus is kept close to the surface of
the intestine by the glycocalyx, which consists of glycoprotein filaments extending from the
interior of absorptive cells to the exterior forming a buffering network above the cell surfaces.
( Å. Krogdahl and A.M. Bakke, 2011). Depending on the species, skin mucus varies
considerably in viscosity, thickness, and glycoprotein (mucin) content which also represents the
major components of mucus (Dash et al., 2018). Mucins are glycoproteins with high molecular
weight, which give the mucus viscoelastic and rheological properties. Other components found
in fish mucus are lysozyme, glycosaminoglycans, immunoglobulins, complements, carbonic
anhydrase, lectins, and calmodulin (Shephard, 1994). However, the composition of the fish skin
mucus is very variable between species and within species, sex, developmental stages, and
environmental conditions (Blackstock and Pickering, 1982; Reverter et al., 2018). In recent
years, interest in antimicrobial properties of fish skin mucus has increased (Kumari et al., 2011;
Reverter et al., 2018), playing an important role against microbes that can alter the mucosal
microbiome of fish, making the fish more vulnerable to several diseases (Llewellyn et al., 2017;
Reid et al., 2017). The antimicrobial effect of mucus appears to be a good candidate for the
development not only of new therapeutic agents to treat infections in humans (Fuochi et al.,
2017) but also of new agents for commercial use (Pethkar and Lokhande, 2017).The fish skin
mucus reported to contain few metabolites having antibacterial properties such as azelaic acid,
N-acetylneuraminic acid and N-acetylglucosamine, and hydroxyisocaproic acid (Ekman et al.,
2015 ▶). Mucus composition varies amongst different fish species. The mucus cells and the
composition of the mucus are influenced by various endogenous (e.g. sex, developmental stage)
and exogenous factors (e.g. stress, hyperosmolarity, pH and infections) (Ellis, 2001 ▶). Mucus is
continuously secreted, then shed and discarded, or digested and recycled. Its lifetime is short,
often measured in minutes to hours, and both the sensory and effector functions mediated in
mucus are rapid; mucus secretion and shedding rates can change within seconds and can increase
dramatically to wash away toxic substances. ( Å. Krogdahl and A.M. Bakke, 2011). Several
studies have shown the potential use of antimicrobial properties of fish skin mucus in the
treatment of infections in humans, and most of them concern freshwater species ( et al., 2009;
Kumari et al., 2011; Nwabueze, 2014; Pethkar and Lokhande, 2017). Mucus acts as a dynamic
physical and biochemical barrier, displaying numerous biological and ecological roles such as
osmoregulation, protection against abrasion, protection against environmental toxins and heavy
metal toxicity, parental feeding, protection against pathogens, and chemical communication. Fish
mucus also contains numerous immune molecules, such as lysozymes, immunoglobulins,
complements, lectins, and antimicrobial peptides (AMPs), and other molecules like
mycosporine-like amino acids (MAAs), toxins, and kairomones—uncharacterized
semiochemicals that mediate interspecific interactions by providing information that benefits
individuals of another species and harms the emitter. Composition of fish mucus and its
rheological properties are vital for the maintenance of mucus functions. Mucus surfaces are
dynamic matrices and their composition varies among fish species and with endogenous (sex and
developmental stage) and exogenous factors (stress, water temperature, pH and infections).
Stress conditions (e.g., handling stress, confinement, food deprivation, exposure to toxic
substances) can change the mucous production and composition (e.g., level of proteins and
immune molecules), compromising fish health and increasing the fish susceptibility tobacterial
pathogens. Mucus viscoelasticity determines its ability to block many types of motilebacteria,
and several studies showed that fish tend to increase their mucus secretion and changetheir
composition when exposed to pathogens, which may contribute to the defence againstthese
pathogens. Furthermore, pathogen infections (e.g., virus, bacteria) can also alter the
mucosalmicrobiome of fish, facilitating the increase of pathogenic bacteria. External fish mucus
is most often sampled by gently scraping (e.g., with cell scraper or spatula) the externalsurfaces
of a fish (body, fins, gills), avoiding ventral areas in order to avoid intestinal or
spermcontaminations. However, a recurrent issue with this method is the possible contamination
ofthe mucus from other tissues such as blood or epithelial cells during fish mucus collection. The
useof adsorbing materials like filter paper or cotton swabs has also been used in fish
mucuscollection as a strategy to avoid epithelial contamination. (Raj et al.) showed that the use
of cottonswabs removed the upper most layers of epidermis, whilst the use of filter paper
allowed mucusremoval without apparent damage of the epithelial cells. A recent study showed
that the metabolomeof mucus samples obtained through absorption displayed the highest
repeatability, whilst those fromscrapped mucus displayed the most variability. Other strategies to
collect fish mucus includerinsing the fish surfaces with different solutions, aspirating the fish
mucus using a vacuumcleaner, and the use of plastic bags filled with different solutions. The
mucus gel matrix is primarily comprised of O-glycosylated proteins (GPs) called mucins,but it
also contains a diverse array of other molecules such as (1) proteins (structural proteins,immune-
related proteins, and antimicrobial peptides and proteins)(reviewed by Brinchmann 2016),(2)
lipids, and (3) smaller molecules such as crinotoxins—fish epidermal toxins not associated with
any venom apparatusand MAAs that display a wide array of biological roles.Fish mucosal
surfaces also harbor a diverse community of organisms (bacteria, fungi, and viruses) that play a
major part in maintaining host health and homeostasis (reviewed by Gomez et al., Llewellyn et
al., and Kelly and Salinas). Since fish mucus is the main barrier against infections,alll its
components (molecules and microbiota) might coordinate to block pathogen entrance by
deploying different antimicrobial activities and participating in immune responses.

Fish mucus layers are the main surface of exchange between fish and the environment,
and they possess important biological and ecological functions. Fish mucus research is
increasing rapidly, along with the development of high-throughput techniques, which allow the
simultaneous study of numerous genes and molecules, enabling a deeper understanding of the
fish mucus composition and its functions. Fish mucus plays a major role against fish infections,
and research has mostly focused on the study of fish mucus bioactive molecules (e.g.,
antimicrobial peptides and immune-related molecules) and associated microbiota due to their
potential in aquaculture and human medicine. However, external fish mucus surfaces also play
important roles in social relationships between conspecifics (fish shoaling, spawning
synchronisation, suitable habitat finding, or alarm signals) and in interspecific interactions such
as prey-predator relationships, parasite–host interactions, and symbiosis. This article reviews the
biological and ecological roles of external (gills and skin) fish mucus, discussing its importance
in fish protection against pathogens and in intra and interspecific interactions. We also discuss
the advances that “omics” sciences are bringing into the fish mucus research and their
importance in studying the fish mucus composition and functions.

(https://www.mdpi.com/348510)

The present study was undertaken to characterize the biochemical composition and
antibacterial activity of skin mucus of fish Hypophthalmichthys nobilis against different human
and fish pathogenic bacterial strains viz. Klebsiella pneumonia, Pseudomonas aeruginosa,
Escherichia coli, Staphylococcus epidermidis, Staphylococcus aureus, Bacillus cereus and
Aeromonas hydrophilla. Methods: Skin mucus of fish H. nobilis was collected by skin scarping
method. Antibacterial activity of mucus extract was carried out by agar well diffusion method
and measured in terms of zone of inhibition(ZOI) in mm. Antibacterial activity of mucus extract
was then compared with two antibiotic amikacin and chloramphenicol. Minimum inhibitory
concentration (MIC) of skin mucus extract was also determined. Results: The biochemical
characterization of epidermal mucus extract revealed the presence of proteins as a major
component (265±2.64 μg/ml) followed by carbohydrate content (63.66±0.88 μg/ml) and lipid
content (0.0077±0.66 g/ml) respectively. Remarkable antimicrobial activity against all the
selected microbial strains was observed. Zone of inhibition (ZOI) shown by crude mucus extract
against all the bacterial strains was found to be significantly higher than higher than
Chloramphenicol. Conclusion: The present study opined that skin mucus of this fish can be used
as potential antimicrobial components.

(https://www.researchgate.net/publication/
304886876_Biochemical_characterization_and_antibacterial_properties_of_fish_skin_muc
us_of_fresh_water_fish_Hypophthalmichthys_nobilis)

2.2 Related Studies

Drier is a kind of deicer that absorbs moisture from atmosphere, and its drying principles
is exactly hydrone to be adsorbed in self the structure or by chemical mode by physics mode to
absorb hydrone and change its chemical constitution, becomes another material.Because it is
inexpensive, moisture sorption effect is good, is widely used in the damp-prrof packing and the
container control wet stock of various material products at present. Furthermore, desiccants can
help to remove moisture from the air if water is an undesirable ingredient in the reaction mixture.
Lithium aluminum hydride and metals such as sodium, as mentioned before, react violently with
water. Water can also affect the weight of an object, for example, a crucible containing a
chemical that must be weighed; a desiccant can help dry out an object to ensure there is no water
left. Desiccants often contain indicator crystals, salts that change color as they absorb water.

According to the study conducted by the inventors of Korea, conventional absorbents are
mainly composed of porous materials such as silica gel and chlorides such as calcium chloride or
magnesium chloride, and silica gel is harmless to the human body, and has the advantage of
regenerating the absorbent by applying heat to the absorbent even after use, in order to solve the
deliquescent properties of the chloride-based, absorbent is mixed/mixed with chloride and porous
silica in order to solve the deliquescent problem of the chloride-based moisture absorbent. In
addition, the shape of the moisture absorbent prepared in accordance with their invention
exhibits a spherical or granular particle size of 0.2 ~ 8mm with an opaque white, and when the
moisture is absorbed, the color of the moisture absorbent is transparent while maintaining the
form of the moisture absorbent. The result of this invention relates to a moisture absorbent which
is easy to be applied to long-term sealed packaging of small expensive products such as
pharmaceuticals or electronic parts, and a manufacturing method thereof. Hygroscopic agents
commonly known as desiccants are currently widely used for the purpose of extending the shelf
life of products by preventing moisture damage by continuously removing moisture while
maintaining freshness and cleanliness in the storage of pharmaceuticals or electronic parts.

According to the study conducted by the researchers of Muntinlupa NHS, they used
organic desiccant to absorb moisture and it was derived from the extraction of slime obtained
from the body secretions of the snail. They used 5.6 grams wet slime and 1.6 grams wet filter
paper; 5.6 grams dry slime and 1.6 grams wet filter paper to observe the difference between
them. One-way Analysis of Variance (ANOVA) for independent groups showed that there was
an overall significant difference among the group means, at P <.0001. The results of post-hoc
Tukey’s HSD tests showed that all points of comparison were significant, P<.01. Their study
proved that the organic desiccant obtained from Snail (Helix aspersa) slime could be effective
moisture absorbent. Moreover, Lauri H Nelson et al. Int J Audio (2017) found that all desiccants
and the white rice were effective in removing moisture from hearing aids, with Hal Hen Super
Dri Aid showing the largest mean reduction in relative humidity. Based on analysis of
covariance results, white rice was statistically similar to several of the commercial desiccants.

According to the study conducted by the doctors of chemical sciences, desiccation or

deep dehydration of liquid organic media is of great importance for modern chemical, gas and
oil-refining industry. The moisture content in organic liquids decreases the operating ability and
reactivity of the product. Furthermore, moisture is released on the walls of pipelines and vessels
when organic liquids are supercooled that causes their corrosion. In general, moisture content in
organic liquids adversely affects the qualitative characteristics of the raw materials and the
products. Limit moisture content in the liquid medium is restrained by solubility, which
decreases with decreasing temperature.

Another study was conducted by Do-Hyung Kim, Seong-NyumJeong and Jae-Hong Lee
by testing the adjunctive use of gel-type desiccating agent for regenerative surgical treatment of
peri-implantitis in patients with inaccessible implant surface. In this study, gel type desiccating
agent was used to disinfect the inaccessible surface of dental implants. Both cases in this report
showed successful clinical and radiographic outcome up to 6 months of follow up.

2.3 Synthesis

According to the study made by Muntinlupa National High School, they extracted slime from
the body of a snail to be used as a desiccant to absorb moisture. They used 5.6 grams wet slime
and 1.6 grams wet filter paper then 5.6 dry slime and 1.6 wet filter paper to observe the
difference. Independent groups showed that there was an overall significant difference among
the group, and all points of comparison were significant. Their study proved that the organic
desiccant from snail(Helix aspersa) slime could be effective absorbent of moisture. According to
the doctors of chemical sciences, desiccation or deep dehydration of liquid organic media is of
great importance for modern chemical, gas and oil- refining industry, much like the study of the
investors from Korea states that porous materials such as silica gel and chlorides such as calcium
chloride or magnesium chloride is harmless to the human body. In addition, the shape of the
moisture absorbent prepared in accordance with their invention, and when the moisture is
absorbed, the color of the moisture absorbent is transparent while maintaining the form of the
moisture absorbent. In general, moisture content in organic liquids adversely affects the
qualitative characteristics of the raw material. Limit moisture content in the liquid medium is
restrained by solubility, which decreases with decreasing temperature.

Drier is a kind of deicer that absorbs moisture from atmosphere, and its drying principles
is exactly hydrone to be adsorbed in self the structure or by chemical mode by physics mode to
absorb hydrone and change its chemical constitution, becomes another material.Because it is
inexpensive,moisture sorption effect is good, is widely used in the damp-proof packing and the
container control wet stock of various material products at present.Furthermore, desiccants can
help to remove moisture from the air if water is an undesirable ingredient in the reaction
mixture.Water can also affect the weight of an object, for example, a crucible containing a
chemical that must be weighed; a desiccant can help dry out an object to ensure there is no water
left. Desiccants often contain indicator crystals, salts that change color as they absorb water.

The present literatures and studies that have been used play a vital role in conducting this
research in the sense of having a guide, different knowledge about the topic, and preserves all the
information needed.
2.4Conceptual Framework

Independent Variable Dependent Variable

Tilapia
Mucus Rate of Absorption of
the Dessicating Agent

Commercialized
Dessicating
Agent

Figure 1: Schematic Diagram: Diagram showing the relationships between the variables used in the study.
The independent variables are both Tilapia mucus and the commercialized dessicating agent in which it
influences or affects the dependent variable which is the rate of absorption of the dessicating agent. The
absorption rate is a great opportunity to showcase or prove the effectiveness of alternative dessicating
agent when tilapia mucus is the main constituent used.