International Journal of Science, Technology, Engineering and Mathematics

Volume 3 Issue 2 June 2023

DOI: https://doi.org/10.53378/352979

Rauvolfia Serpentina and Peperomia

Pellucida as Antiparasitic Spray Against
Rhipicephalus Sanguineus Latreille
1
Mahmooda Aziza Bhatti, 2Mutahir Saeed, 3Marites Hugo &
4
Jerico Guevarra

Abstract

Rauwolfia serpentina (serpentina) and peperomia pellucida (pansit-pansitan) were used as the raw materials for
creating different mixtures to determine which concentration is faster in exterminating Rhipicephalus
sanguineus latrille (common ticks). The process used was decoction, the harvested extract was used for the
creation of the spray mixtures. The process used three different preparations of mixture concentration with
variants concentration ratio of serpentina/pansit-pansitan as: mixture 1 with 50%/50%; mixture 2 with
75%/25%, and mixture 3 with 25%/75%. The study recorded the testing times in exterminating the ticks as:
mixture 1 had the results in 1.5 hours (sprayed once), 1.33 hours (sprayed twice), and 1.25 hours (sprayed
thrice); mixture 2 had the results in 3.5 hours (sprayed once), 3.42 hours (sprayed twice), and 3.33 hours
(sprayed thrice); and mixture 3 had the results in 1.5 hours (sprayed once), 1.42 hours (sprayed twice), and 1.33
hours (sprayed thrice). The experiment showed that mixture 1 and mixture 3 almost had the same results, if not
for a few second differences, mixture 1 was always faster than mixture 3, while mixture 2 was left behind for a
few hours. Therefore, mixture 1 is the best concentration to exterminate the ticks in a controlled environment.
Keywords: Serpentina, Pansit-Pansitan, Ticks, Antiparasitic

Article History:
Received: February 21, 2023 Revised: March 24, 2023
Accepted: March 28, 2023 Published online: April 7, 2023

Suggested Citation:
Bhatti, M., Saeed, M., Hugo, M. & Guevarra, J. (2023). Rauvolfia Serpentina and Peperomia Pellucida as
Antiparasitic Spray Against Rhipicephalus Sanguineus Latreille. International Journal of Science, Technology,
Engineering and Mathematics, 3 (2), 1 - 19. https://doi.org/10.53378/352979
About the authors:
1Corresponding author. An alumna of City of San Jose del Monte National Science High School and currently taking up
Bachelor of secondary Education Major in English Minor in Mandarin at Bulacan State University. Corresponding email:
azizahbhatti@gmail.com
2Graduate of Biology, Naturopathy Health Science, and Alternative Medicine. Worked as an Associate Researcher under the

Department of Health on Herbal and Natural Medicine.

3Teacher 3 at City of San Jose del Monte National Science High School and Junior High School department coordinator.

Currently pursuing her Masters degree in Educational Managaement at Polytechinic University of the Philippines.
4Special Science Teacher I at City of San Jose del Monte National Science School and currently the Senior High School research

coordinator. Pursuing Masters Degree in Education Major in Chemistry at Bulacan State University.
* This paper is a finalist in the International Research Competition 2022 Category 1 – High School.

© The author (s). Published by Institute of Industry and Academic Research Incorporated.
This is an open-access article published under the Creative Commons Attribution (CC BY 4.0)
license, which grants anyone to reproduce, redistribute and transform, commercially or non-
commercially, with proper attribution. Read full license details here:
https://creativecommons.org/licenses/by/4.0/.
2 | International Journal of Science, Technology, Engineering and Mathematics, Volume 3 Issue 2

1. Introduction

Rauwolfia serpentina has been used since the pre-vedic period to treat many
infections and diseases. It is a large glabrous herb or shrub belonging to the family
Apocynaceae and found in Assam, Pegu, the Himalayas, Java, Tenasserim, Deccan,
Peninsula, Bihar, and the Malay Peninsula. It is a source of many phytoconstituents including
alkaloids, carbohydrates, flavonoids, glycosides, phlorotannins, phenols, resins, saponins
sterols, tannins, and terpenes (Chauhan et al., 2017). In a study by members of the
Department of Chemistry at HNB Garhwal University, they used the roots of Rauwolfia
Serpentina against Salmonella Typhimurium, Escherichia Coli (E-Coli), Citrobacter freundii,
Proteus Vulgaris, Enterococcus faecalis, and Staphylococcus Aureus. The researchers have
commented that “the research supports folklore” (Negi et al., 2014). On the other hand,
pansit-pansitan, scientifically known as peperomia pellucida, is a medicinal plant with anti-
gout properties. Its anti-gout properties help lower the uric acid amount in the blood. It was
one of the ten medicinal plants which underwent clinical testing as per orders of the
Department of Health (DOH).

A study by Akinnibosun et al. (2021) on the antibacterial activity of pansit-pansitan

against three-gram negative bacterial isolates showed that the plant indeed has antibacterial
properties. The plant undergoes a decoction process, and its extract was added to solvents,
namely water, and ethanol. The results varied with the two solvents, wherein peperomia
pellucida with ethanol is much more effective than peperomia pellucida with water. While
the study focused only on bacteria and viruses using pansit-pansitan and serpentina
separately, this study investigated the possibility of mixing the two (2) herbal medicines and
finding out if they have powerful and effective effects in exterminating ticks specifically the
brown ticks. Brown ticks, scientifically referred to as rhipicephalus sanguineus latrille, is a
common tick found almost in any dog. This type of tick is usually reddish-brown in their
adulthood, with no specific marks, unlike other species. Brown dog ticks often travel into
houses on canines, their preferred hosts, or cats. Because they are found deep within the hair
of animals, homeowners may not immediately see them. Adult ticks typically embed
themselves to a dog’s ears and between its toes, while larvae and nymphs typically attach to
the back of the dog. If brown dog ticks do not have a host to feed upon, they will readily seek
 ISSN 2799-1601 (Print) 2799-161X (Online) | 3

out humans for them to survive. The pests attach themselves to an animal's skin to feed on its
blood and lay eggs in its fur. After entering homes, they breed and can spread onto residents
and other pets (Orkin, 2021).

This study aims to concoct a substance made from serpentina and pansit pansitan that
exterminates the brown ticks among the animals. This uses variations of solutions and
durations of tick exposure to the mixture. Although these plants have antibacterial properties
and ticks are not bacteria, the study used these plants to remove ticks found on domestic cats
and dogs, to provide alternative herbal antiparasitic spray without harmful chemicals thus
lessening the environmental hazard and health hazard for both animals and human.

2. Literature review
2.1. Serpentina

The existence of enormous therapeutic properties makes Rauvolfia serpentina an

essential medicinal plant in the pharmaceutical world (Khurshid et al., 2022). Because of the
inclusion of alkaloids, carbohydrates, flavonoids, glycosides, phlobatannins, phenols, resins,
saponins, sterols, tannins, and terpenes, the plant is used to treat a variety of ailments
(Malviya & Sason, 2016). High blood pressure (Lobay, 2015), emotional agitation, epilepsy,
traumas, anxiety, excitement, hysteria, sedative insomnia, and insanity (Ali et al., 2022) have
all been treated with plant bits, seeds, and rhizomes for centuries in Ayurvedic medicine
(Khurshid et al., 2022).

Rauwolfia Serpentina is noted to have reserpine, a substance that is used to cure

hypertension (Lobay, 2015). In a study by the Department of Chemistry at HNB Garhwal
University, they used the roots of Rauwolfia Serpentina against Salmonella Typhimurium,
Escherichia Coli (E-Coli), Citrobacter freundii, Proteus vulgaris, Enterococcus faecalis, and
Staphylococcus Aureus to quantify the reserpine content of the plant and test the
antimicrobial effectiveness of its menthol extract. The menthol extract of Serpentina
inhibited the growth of several bacterial species and concluded that the study supports the
folklore claims of the plant species (Negi et al., 2014). Rauwolfia serpentina has been used
since the pre-vedic period for the treatment of a lot of infect more than 50 distinct alkaloids
in the plant. Ajmaline, ajmalicine, ajmalimine, deserpidine, indobine, indobinine, reserpine,
reserpiline, rescinnamine, serpentine, and yohimbine are the most important alkaloids.
4 | International Journal of Science, Technology, Engineering and Mathematics, Volume 3 Issue 2

Antimicrobial, antifungal, anti-inflammatory, antiproliferative, antidiuretic, and

anticholinergic properties are also known for serpentina. Because of its societal acceptability,
greater compliance with the human body, and fewer side effects, herbal medicine is now the
source of primary health care for 75–80 percent of the world's population (Ekor, 2014; Welz
et al., 2018). As a result, naturally occurring remedies are now alternatives to save millions
of patients around the world. The current study attempts to examine the different
pharmacological, phytochemical, and medicinal effects of serpentina as a result of both of
these properties (Kumari et al., 2013).

In a study, Alshahrani et al. (2021) explored the antibacterial activity of R. serpentina.

Ethanolic extract of root was evaluated using the well-diffusion method. Two Gram-positive
(Bacillus subtilis and Staphylococcus) and three gram-negative bacteria (Klebsiella
pneumoniae, Pseudomonas aeruginosa, and Salmonella typhimurium) were used for the
activity of which only three bacteria Klebsiella pneumonia, Staphylococcus, and B. subtilis
bacteria are found susceptible. Similarly, Negi et al. (2014) studied the antibacterial activity
of methanolic extract of roots (MREt) of R. serpentina. Antibacterial activity was evaluated
using the agar well diffusion method against gram-positive and gram-negative bacteria for
the determination of minimum inhibitory concentration (MIC) and the diameter of the zone
of inhibition (ZOI). The study revealed that Staphylococcus aureus shows the highest ZOI
(13 mm) with the lowest MIC (625 µg) and Escherichia coli possess the highest MIC (10
mg), whereas Proteus vulgaris was observed resistant to tested extracts up to 10 mg. Hence,
R. serpentina exhibited strong antibacterial activity. Nigussie et al. (2021) used methanolic
and chloroform extracts of the leaf and root of R. serpentina for antibacterial activity. The
activity was assessed against S. aureus, E. coli, P. aeruginosa, B. subtilis, and K. pneumonia
by disk diffusion method. 50 μl/ml concentrations of leaf and root chloroform extracts
showed no ZOI against S. aureus and B. subtilis. Maximum zone inhibition was observed at
15.0 mm and 15.5 mm against E. coli for leaf and root extract, respectively. 100 μl/ml
concentration showed maximum zone inhibition against all test organisms for both leaf and
root extract. All the bacteria were more susceptible to methanolic extract than chloroform (
(Chauhan et al., 2017).

Azmi and Qureshi (2012) determined the phytochemistry and effect of Rauwolfia
serpentina methanolic root extract (MREt) on diabetic alloxan-induced male mice. Mice were
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categorized into diabetic (distilled water at 1 mL/kg), negative (0.05 percent dimethyl
sulfoxide at 1 mL/kg), positive (glibenclamide at 5 mg/kg) controls, and three test classes
(MREt at 10, 30, and 60 mg/kg). For 14 days, all medications were given orally. MREt
included alkaloids, carbohydrates, flavonoids, glycosides, cardiac glycosides, phlobatannins,
resins, saponins, hormones, tannins, and triterpenoids qualitatively, while the extract
contained complete phenols quantitatively. Root powder was also tested for flavonoids,
saponins, and alkaloids. When opposed to diabetic treatment, MREt was shown to be
successful in increasing body weights, glucose and insulin levels, insulin/glucose ratio,
glycosylated and total hemoglobin in research groups. Total cholesterol, triglycerides, low-
density lipoprotein (LDL-c), and relatively low-density lipoprotein (VLDL-c) cholesterol
levels were all found to be significantly lower in the test groups. Both research groups' liver
tissues showed significant lipolysis and increased glycogenesis. All of the groups had
standard ALT levels. In alloxan-induced diabetic mice, MREt increases glycemic,
antiatherogenic, coronary risk, and cardioprotective indices.

In another study, Santhosh et al. (2016) analyzed antibacterial activity and

preliminary phytochemical screening of Endophytic Fugal Extract of Rauvolfia serpentine.
Sarpgandha (Apocynaceae) is a common medicinal plant that is best known for its numerous
phytochemicals. The key goal of this research was to see whether Rauvolfia serpentina L.
had some antifungal action against Alternaria alternata, Aspergillus flavus, and Mucor rouxii,
which are all phytopathogenic fungi. The antifungal function of aqueous extracts of the entire
plant, stem, and roots of Rauvolfia serpentina L. was investigated using an agar well
diffusion assay. Rauvolfia serpentina L. aqueous root extract had significantly greater
antifungal efficacy than the other extracts tested against Alternaria alternata and Aspergillus
flavus. Their research explicitly demonstrates Rauvolfia serpentina L.'s antifungal properties,
implying that it may be used to combat pest control in a variety of plants and animals.
Endophytic fungi isolated from Rauvolfia serpentina, a well-known Indian medicinal herb,
are used in Ayurveda for the treatment of many diseases (Santhosh et al., 2016). The
antibacterial behavior of isolated endophytes against pathogenic bacteria was tested. Twenty
fungal isolates were recovered from various sections of the host plant and characterized for
their morphological features using Scanning Electron Microscopy (SEM). They were
classified into eight genera based on observations: Fusarium sp., Phomopsis sp.,
6 | International Journal of Science, Technology, Engineering and Mathematics, Volume 3 Issue 2

Colletotrichum sp., Cladosporium sp., Aspergillus sp., Xylaria sp., Alterneria sp., and
Gleomastix sp. The examination of the extract against the target bacteria exposed the secret
of the medicinal plant's fungal endophytes. Colletotrichum sp. (Rs-R5), Fusarium sp. (Rs-
R1), (Rs-R7), and Cladosporium sp. (Rs-S4) extracts were shown to be selective against
human pathogenic bacterial strains E. coli (ATCC 25922), Gram-negative bacteria, and S.
aureus (ATCC 25323), Gram-positive bacteria. The most effective sample was an ethyl
acetate extract of an active fungal isolate (Colletotrichum sp; Rs-R 5) against E. coli and S.
aureus, with maximal inhibition zones of 16 mm and 14 mm and minimum MICs of 25 g/ml
and 36.5 g/ml, respectively. They found eight endophytic fungal genera in R. serpentina,
according to Santhosh et al., (2016): Fusarium sp., Alternaria sp., Phomopsis sp., Xylaria sp.,
Gleomastix sp., Aspergillus sp., Cladosporium sp., and Colletotrichum sp. Out of 20 fungal
isolates tested, four showed antibacterial activity: Fusarium sp. (Rs-R1, Rs-S7),
Cladosporium sp. (Rs-R5), and Colletotrichum sp. (Rs-R5). Using ethyl acetate extract, the
inhibition zone and MIC were detected. Against E. coli, the maximal inhibition region (16
mm) and minimum MIC (25 g/ml) were observed. Secondary metabolites such as alkaloids,
polyphenols, flavonoids, hormones, and saponins were present in abundance in the ethyl
acetate sample.

Plant products are gaining popularity as bactericides and fungicides (Sharanabasappa

et al., 2015) due to their systemic efficacy and low phototoxicity. Sharanabasappa et al.
(2015) attempted the antibacterial and pharmacological effects of various Rauvolfia
serpentina extracts since a significant number of plants are recognized for their antibacterial
and antifungal function. They tested the antibacterial efficacy of various Rauvolfia serpentina
extracts against E. coli, Klebsiella, Pseudomonas, and S. Aureus. The components used are
as follows: Nutrient agar medium, sterile Petri dishes, 0.1–0.2ml pipettes, cultures, nutrient
broth, and sterile test tubes containing a proven concentration of the extract of the solutions.
The cup plate method was used in the experiments, with a dosage of 1 mg/ml. Water and
chloroform extracts were the most effective against bacterial strains E. coli and S. aureus. E.
coli and P. aeruginosa Klebsiella and the rest of the Pet ether and ethanol extracts were also
effective against P. aeruginosa and S. aureus. Various Rauvolfia serpentina extracts were
examined for antifungal activity among various extracts of the Pet. Ether and ethanol extracts
were found to be effective against A. flavus and A. niger. The operation of niger and the
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remaining extracts was low to moderate. The antifungal efficacy of plant extracts was
compared to that of normal antifungal drugs fluconazole by cup plate process, with A.
Flavus and A. niger as the fungi chosen for this. Rauvolfia serpentina extracts were tested for
antibacterial and antifungal function. Rauvolfia serpentina extracts were tested for
antibacterial and antifungal function. The water and chloroform extracts were the most
effective against the bacterial strains E.coli and P. klebsiella, as well as the remaining Pet.
ether and ethanol extracts were also effective against P. aeruginosa and S. aureus. The Pet.
Ether and Ethanol extracts had excellent antifungal activity against A. flavus and A. niger,
while the other extracts had low to moderate antifungal activity (Sharanabasappa et al.,
2015).

2.2. Pansit-Pansitan

Pansit-pansitan, scientifically known as peperomia pellucida, is a medicinal plant

valued for its anti-gout properties which help lower uric acid in the blood. It was one of the
ten clinically tested medicinal plants endorsed by the DOH. It was reported that pansit-
pansitan contained high amounts of toxic metals like lead (Pb) and cadmium (Cd) surpassing
the limits allowed by the World Health Organization (WHO). To regulate the uptake of
nutrient elements in pansit-pansitan, the use of hydroponic culture through non-aerated
Hoagland's solution was studied (De Guzman, 2000).

Peperomia pellucida (Linn.) is a piperaceae bush with a glossy or silvery appearance.

This plant's ethnomedicinal applications include curing stomach pain, abscesses,
inflammation, boils, colic, and exhaustion (Gomes et al., 2022). Abdulrazaq (2018) extracted
from the air-dried leaves of P. pellucida the essential oils using a Clevenger apparatus and a
hydro distillation process. The essential oil obtained was light yellow in color, had an
unpleasant odor, and yielded 0.30 v/w. Elemol (9.32 percent), Neointermedeol (8.35
percent), 1H-3a,7-Methanoazulene- (5.60 percent), and Bicyclo [2.2.1] heptanes,2,2,3-
trimethyl are the main constituents (5.08 percent). A large amount of the leaf oil was made
up of sesquiterpenes and oxygenated sesquiterpenes (52.71 percent). The antimicrobial
effects revealed that it has a Minimum Inhibition Concentration (MIC) on Pseudomonas
aeruginosa and Bacillus subtilis at 0.01 percent oil concentration, with inhibition zones of 7.0
mm and 9.3 mm, respectively; however, MIC against Bacillus cereus was obtained at 0.1
8 | International Journal of Science, Technology, Engineering and Mathematics, Volume 3 Issue 2

percent oil concentration. Antifungal tests on Lasiodiplodia theobromae, Fusarium

oxysporum, and Aspergillus tamari at percent levels that revealed the plant's essential oils
had potent antifungal effects on all three fungi species. These findings suggested that the
plant's essential oil could be used as an antimicrobial agent (Abdulrazaq, 2018).

The anticancer, antimicrobial, antioxidant, and chemical compositions of Peperomia

pellucida leaf extract were also studied. In the study of Wei et al. (2011), the anticancer
activity of P. pellucida leaf extract was determined using a colorimetric MTT (tetrazolium)
assay against the human breast adenocarcinoma (MCF-7) cell line, and the plant extract's
antimicrobial property was discovered using a two-fold broth microdilution system against
10 bacterial isolates. The plant extract's antioxidant activity was then determined using the
DPPH radical scavenging process, and the chemical compositions were screened and
classified using gas chromatography-mass spectrometry (GC-MS). The findings of this
analysis revealed that P. pellucida leaf extract had anticancer properties, with an IC50 of
10.4±0.06 g/ml. The plant extract was found to inhibit the growth of Edwardsiella tarda,
Escherichia coli, Flavobacterium sp., Pseudomonas aeruginosa, and Vibrio cholerae at 31.25
mg/l; Klebsiella sp., Aeromonas hydrophila, and Vibrio alginolyticus at 62.5 mg/l; and
Salmonella sp. and Vibrio parahaemolyticus at 125 mg/l. The plant extract was observed to
inhibit 30% of DPPH, a free radical, at a concentration of 0.625 ppt. The main compound in
the plant extract was phytol (37.88%), followed by 2-Naphthalenol, decahydro- (26.20%),
Hexadecanoic acid, methyl ester (18.31%), and 9,12 Octadecadienoic acid (Z, Z)-, methyl
ester (9.12%). (17.61 percent). The results of this study showed that a methanol extract of P.
pellucida leaf had a lot of potential as a medicinal drug, particularly in the treatment of breast
cancer (Wei et al., 2011).

In the study of Bojo et al. (1995), fresh and air-dried Peperomia pellucida plants were
subjected to a differential extraction method using three solvents: methanol-water (14:1),
ethyl acetate, and hexane. The ethyl acetate extract yielded a strong antibacterial extract. A
major fraction of the ethyl acetate extract was shown to have strong antibacterial efficacy
against Staphylococcus aureus, Bacillus subtilis, and Pseudomonas aeruginosa that was more
active than the penicillin norm, indicating its ability as a wide spectrum antibiotic.
Meanwhile, the study of Apatas et al. (2020) aimed to see whether Pansit-pansitan
(Peperomia pellucida Linn) aqueous leaf extract has anti-inflammatory properties in vitro.
 ISSN 2799-1601 (Print) 2799-161X (Online) | 9

Fresh leaves were gathered, air dried, and aqueous extracted before being prepared in various
doses (200, 400, 600, 800, and 1000ug/mL). In-vitro anti-inflammatory activity was assessed
using inhibitors of HRBC lysis and protein denaturation. The existence of secondary
metabolites, which are believed to have anti-inflammatory properties, may be due to the
aqueous extract's anti-inflammatory effect on the plant. However, these metabolites are not
sufficient enough when used in low dosages.

2.3.Ticks

Rhipicephalus sanguineus is usually reddish brown in their adulthood, with no

specific marks, unlike other species. Brown dog ticks often travel into houses on canines,
their preferred hosts, or cats. Because they are found deep within the hair of animals,
homeowners may not immediately see them. Adult ticks typically embed themselves in a
dog’s ears and between its toes, while larvae and nymphs typically attach to the dog's back.
If brown dog ticks do not have a preferred host to feed upon, they will readily seek out
humans for their needed blood meals. The pests attach themselves to an animal's skin to feed
on its blood and lay eggs in its fur. After entering homes, they breed and can spread to
residents and other pets. Unlike other species, brown dog ticks can survive and complete
their entire life cycle indoors. Warm temperatures help these pests develop and reproduce,
causing infestations to spread quickly. Brown dog ticks may transmit canine-related diseases,
such as canine ehrlichiosis and babesiosis. They are known transmitters of Rocky Mountain
spotted fever to humans, but are not known to transmit Lyme disease. However, they are
often mistaken for deer ticks, which are known carriers of Lyme disease (Orkin, 2021).

According to John et al. (2017), ticks are tiny crawling bugs in the spider family that
feed by sucking blood from animals. They are second only to mosquitoes as vectors of
human disease, both infectious and toxic. Infected ticks spread over a hundred diseases, some
of which are fatal if undetected. They spread the spirochete (which multiplies in the insect's
gut) with a subsequent bite to the next host. Among vector-borne diseases, the most common,
Lyme disease, also known as the great mimicker, can present with rheumatoid arthritis,
fibromyalgia, depression, attention deficit hyperactivity disorder, multiple sclerosis, chronic
fatigue syndrome, cardiac manifestations, encephalitis, mental illness, name some of the
many associations.
10 | International Journal of Science, Technology, Engineering and Mathematics, Volume 3 Issue 2

3. Methodology

This experimental study strictly followed scientific procedure. The series of steps
were followed to generate accurate and substantial data collection.

1. Prepare and gather all materials that are required in the whole procedure of
making the antibacterial spray.
2. Wash all the leaves with tap water.
3. Crush 20 leaves with a mechanical blender to get their extract.
4. Boil each 20 crushed leaves in separate pots with 400 ml water.
5. Stir the materials evenly for 30 seconds up to 1 minute.
6. Prepare 3 glass containers and label them from mixtures 1 to 3. Use the funnel
and measuring cups to follow the percentages needed for different concentrations
of the mixtures.

Mixture 1: 200 ml of Serpentina and 200 ml of Pansit-pansitan Mixture

Mixture 2: 300 ml of Serpentina and 100 ml of Pansit-pansitan Mixture.

Mixture 3: 100 ml of Serpentina and 300 ml of Pansit-pansitan.

7. Stir the materials evenly for 30 seconds up to 1 minute.

8. Cool the mixture and transfer it to a spray bottle.
9. Prepare 9 plastic cups and fill them with 3 ticks each with the following label:

Label 1 - Mixture 1 (spray once)

Label 2 - Mixture 1 (spray twice)

Label 3 - Mixture 1 (spray thrice)

Label 4 - Mixture 2 (spray once)

Label 5 - Mixture 2 (spray twice)

Label 6 - Mixture 2 (spray thrice)

Label 7 - Mixture 3 (spray once)

Label 8 - Mixture 3 (spray twice)

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Label 9 - Mixture 3 (spray thrice)

10. Every solution is sprayed in the labeled plastic cups of different concentration
11. After an hour from the first spray, record how many ticks were found dead.
12. This manner of checking is done until all the ticks are dead.

The duration of the experiment is as follows:

Week 1. The last days of week 1 will be fully devoted to the creation of the mixture.
To be safe, the study used two (2) days for the preparation of the mixture.

Week 2. This is the period of experimentation and observation. Every hour, the
researcher sprays the ticks in the petri dish with the mixture assigned to them. The
experiment and observation are enclosed in a week.

The method of observation is descriptive, wherein the ticks were placed in a

controlled environment and then exposed to the mixture of serpentina and pansit-pansitan in
different amount of concentrations. It was observed for an hour and the observer took notes
of all the changes that took place.
4. Findings and Discussion

This study experiments on Rauvolfia Serpentina (Serpentina) and Peperomia

pellucida (Pansit-Pansitan) as an anti-parasitic spray against Rhipicephalus Sanguineus
Latreille (Common Ticks). The first scientific query is the time required for the anti-
parasitic spray treatment to suppress the ticks as shown in table 1.

Table 1

Testing results of the mixtures

Mixture 1 Mixture 2 Mixture 3

Once 1hr 30mins (1.5 hrs) 3hrs 30mins (3.5 hrs) 1hr 30mins (1.5 hrs)

Twice 1hr 20mins (1.33 hrs) 3hrs 25mins (3.42 hrs) 1hr 25mins (1.42 hrs)

Thrice 1hr 15mins (1.25 hrs) 3hrs 20mins (3.33 hrs) 1hr 20mins (1.33 hrs)
12 | International Journal of Science, Technology, Engineering and Mathematics, Volume 3 Issue 2

The table shows the number of times that the mixture has been sprayed on the ticks in
a controlled environment. It also shows the length of time that the ticks exterminate after the
mixture has been sprayed with Mixture 1 with the shortest period in the three instances of
spraying. The longest period of extermination is Mixture 2 with minimum of 3 hours 20
minutes in three sprays.

Table 2

The number of ticks that exterminate

Mixture Mixture 1 Mixture 2 Mixture 3

Number of times that the mixture has been sprayed 3 3 3
Several ticks died 3 2 2

Table 2 shows the number of times that the mixture has been sprayed on the ticks. In
a span of 1 (one) hour and 30 (thirty) minutes, mixture 1 killed 3 ticks and mixture 2 killed 2
ticks.

The experiment used three solutions or three variants of concentration. The three
mixtures used are: mixture 1 has 50%/50% of Serpentina and Pansit-Pansitan diluted with
water; mixture 2 has 75%/25% of Serpentina and Pansit-Pansitan diluted with water; and
mixture 3 has 25%/75% of Serpentina and Pansit-Pansitan diluted with water. The study
experimented on the appropriate mixture for extermination with results shown in table 3.

Table 3

Summary of testing results

Mixture 1 Mixture 2 Mixture 3

Number Number Number Number

Times Number Times Times it Number
of ticks of of ticks of ticks
sprayed of Hours sprayed sprayed of Hours
died Hours died died

1 1.5 3 1 3.5 2 1 1.5 2

2 1.33 3 2 3.42 2 2 1.42 2

3 1.25 3 3 3.33 2 3 1.33 2

Average 1.36 3 2 3.42 2 2 1.42 2

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Table 3 shows the summary of the testing of the three (3) mixtures of serpentina and
pansit-pansitan in different concentrations spraying against the brown ticks in a controlled
environment. For mixture 1, the average time in hours that the brown ticks have been
exterminated is 1.36 hours in relation to the number of times the mixture is sprayed. The
brown ticks that have been killed are 3 from one (1) to three (3) sprays of the mixture. In
mixture 2, the average time in hours that the brown ticks have been exterminated is 3.42
hours and it only kills 2 ticks with the different number of times that the mixture is sprayed.
Lastly, mixture 3 has an average of 1.42 hours in relation to the number of times that the
mixture was sprayed on the brown ticks. It exterminated 2 ticks in each number of times that
the mixture was sprayed.

Table 4

Summary Result of the Three (3) Mixtures

Groups Count Sum Average Variance

Mix1 3 6.66 2.22 0.04120

Mix2 3 1.75 0.58 0.00023

Mix3 3 4.24 1.41 0.00723

Table 4 shows the number of times that the mixtures are sprayed on the brown ticks
and the average time, in an hour, that the mixture exterminated the ticks. It also shows the
computed variance of each mixture.

Table 5

Result of One-Way ANOVA

Source of Variation SS df MS F P-value F crit

Between Groups 4.018289 2 2.009144 123.8514 0.0000132 5.143253

Within Groups 0.097333 6 0.016222

Total 4.115622 8

* Significant difference among groups (P-value < 0.05)

14 | International Journal of Science, Technology, Engineering and Mathematics, Volume 3 Issue 2

Table 5 shows the result of the one-way analysis of variance (ANOVA). It shows the
degree of freedom (df) which are 2 for between groups and 6 for within the groups, then the
calculated F value which is 123.8514 along with the P-value of 0.0000132 while the critical
F value is 5.143253. For the p-value, it is 0.0000132 in comparison to the level of significant
value which is 0.05 implying significant difference in the mixtures.

Table 6

t-test Results between Mixtures

Between Mixtures 1 & 2 Between Mixtures 1 & 3

Mix1 Mix2 Mix1 Mix3
Mean 2.22 0.583333 2.22 1.413333
Variance 0.0412 0.000233 0.0412 0.007233
Observations 3 3 3 3
Hypothesized Mean Difference 0 0
Df 2 3
t Stat 13.92663 6.348667
P(T<=t) one-tail 0.002558 0.003953
t Critical one-tail 2.919986 2.353363
P(T<=t) two-tail 0.005116 0.007906
t Critical two-tail 4.302653 3.182446

Table 6 shows the t-test between mixture 1 and mixture 2. The mean of mixture 1 is
2.22 while mixture 2 is 0.58333 the calculated t-value is 13.92663 against the critical t-value
is 2.919986 for one-tail and 4.302653 for two-tail and the degree of freedom (df) value is 2.
Meanwhile, the result of t-test between mixture 1 and mixture 3 shows the mean for mixture
1 is 2.22 and for mixture 3 it is 1.413333, the degree of freedom value is 3. The calculated t-
value is 6.348667 while the critical t-value for one-tail is 2.353363 and for two-tail is
3.182446.

Table 6

The Average Number of Ticks that Died per Hour

Mix1 Mix2 Mix3

2.00 0.57 1.33

2.26 0.58 1.41

2.40 0.60 1.50

 ISSN 2799-1601 (Print) 2799-161X (Online) | 15

Table 6 shows the average number of ticks died per hour after the mixture have been
sprayed on the brown ticks. Mixture 1 had exterminated an average of two (2) ticks for an
hour followed by mixture 2 with less than 1 tick exterminated and mixture 3 with an average
1 tick per hour.

After all the testing and calculations, the most effective variation in concentration out
of the 3 mixtures used against Rhipicephalus Sanguineus Latreille is mixture 1, which
consists of 50%/50% of Serpentina and Pansit-Pansitan diluted with water. It showed the
fastest effect in suppressing Rhipicephalus Sanguineus Latreille with the shortest amount of
time needed. As shown in the result in table 6, mixture 1 has exterminated an average of 2
brown ticks compared to the other two mixtures which exterminated less than 1 tick for
mixture 1 and 1.5 ticks for mixture 3.

The results have also been confirmed using the one-way ANOVA. In comparison to
the level of significance value of 0.05, the calculated p-value is 0.0000132 which is less than
0.05 and means that there is a significant difference among the groups of mixture that have
been used. This only signifies that the most effective concentration of the mixture is 50%
pansit-pansitan and 50% serpentina.

Further confirmation of the result, the study used the t-test between mixture 1 and 2
which shows that mixture 1 is still the most effective concentration having the t-value of
13.92663 which is way up higher than the critical t-value of 2.919986 for one-tail and
4.302653 for two-tail. Thus, the result shows that there is a significant difference between
mixture 1 and mixture 2. In comparing mixture 1 to mixture 3, another t-test have been done
to prove that mixture 1 is the most effective concentration. The calculated t-value is
6.348667 which is also higher than the critical t-value having 2.353363 for one-tail and
3.182446 for two-tail. The p-value also confirms the result, having the following value:
0.003953 for one-tail and 0.007906 in comparison to a 0.05 level of significance.

The results further showed that the most effective application out of the 3 durations
provided against Rhipicephalus Sanguineus Latreille is thrice per hour. It showed the
shortest amount of time in suppressing the tick in all three mixtures used. The result of the
experimentation showed that Rauvolfia Serpentina (Serpentina) and Peperomia Pellucida
(Pansit-Pansitan) as an antiparasitic spray against Rhipicephalus Sanguineus Latreille
16 | International Journal of Science, Technology, Engineering and Mathematics, Volume 3 Issue 2

(Common Ticks) is effective.

5. Conclusion

This study proved that the mixture of pansit-pansitan and serpentina can really
exterminate the brown ticks as a result of physical observation and statistical analysis.
Statistically, the mixture that has the most effective concentration in exterminating the
brown ticks is mixture 1 for it shows that it can exterminate a minimum of 2 brown ticks in
an hour compared to the other concentration can only exterminate less than 1 tick and 1.5
ticks for mixture 2 and 3 respectively in an hour. Even though all mixtures are effective in
exterminating ticks, mixture 2, which has a 75/25 ratio of Serpentina and Pansit-pansitan, is
the slowest followed by the mixture 3. Therefore, mixture 1 is the fastest option in terms of
effectiveness and rapidity in the extermination of ticks.

6. Acknowledgment

This paper is a school research requirement of the corresponding author. Hence,

sincere gratitude is extended to Ms. Marites Hugo, research adviser, and Mutahir Saeed
Bhatti, father.

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