RESEARCH GRADE 10 QUARTER 1

MODULE # 1
Writing of a revised Research
Proposal
Directions: Given the checklist of criteria below, study and evaluate the
given Research Plan below by checking the box corresponding to your
answer.
Submission Date: Nov. 6 & 7, 2023. This is an individual output and submission.

RESEARCH PLAN
In vitro Antihelmintic Activity of Lantana camara
Extracts on Eudrilus euginiae

Life Science

A. Rationale

Soil-transmitted helminths (STHs) comprise a number of intestinal nematodes

i.e. hookworms (Ancylostoma duodenale, Necator americanus), roundworm (Ascaris
lumbricoides), whipworm (Trichuris trichiura), and Strongyloides stercoralis
according to the World Health Organization (WHO). In the Philippines, these
diseases are key contributing factors to the prevalence of malnutrition, anemia,
eosinophilia and pneumonia .
The emergence of anthelminthic resistance against virtually all available
commercial drugs has posed serious impediments to the effective control of
helminth infestations (Sondhi et al., 1994; Waller and Thamsborg, 2004), hence
the increasing demand towards natural anthelmintics and tendency towards
medicinal plants as alternative anthelmintics.
Lantana camara is an erect shrub widely cultivated in tropical Asia including
the Philippines. Various parts are widely used in folk medicine (Dugasani et al.,
2010; Rangasamy et al., 2010; Aderogba et al., 2008; Devaki et al., 2011; Kannan
et al., 2010).
According to various studies, in vitro antihelminthic activities may be evaluated
on Eudrilus euginiae, as model organism due to its anatomical and physiological
resemblance with intestinal roundworm parasites of human beings (Deore et al.,
2009; Pueblos et al., 2015; James and Ojonugwa, 2014).

B. Research Question, Hypothesis, Scientific Goals, Expected Outcome

Research Question
1. Will Lantana camara leaves, young stem, and flower aqueous extract exhibit in
vitro anthelmintic activity against Eudrilus euginiae?
2. Is there a significant difference on the in vitro anthelminthic activity of varying
concentrations of Baho-baho (Lantana camara) aqueous extract using the
leaves, young stem, and flower in terms of the worms’ paralysis time?
3. Is there a significant difference on the in vitro anthelminthic activity of varying
concentrations of Baho-baho (Lantana camara) aqueous extract using the
leaves, young stem, and flower in terms of the worms’ lethal time?

Null Hypotheses:
1. Baho-baho (Lantana camara) aqueous extract using the leaves, young stem, and
flower will not exhibit in vitro anthelmintic activity against Eudrilus euginiae.
2. There is no significant difference on the in vitro anthelminthic activity of
varying concentrations of Baho-baho (Lantana camara) aqueous extract using
the leaves, young stem, and flower in terms of the worms’ paralysis time..
3. There is no significant difference on the in vitro anthelminthic activity of
varying concentrations of Baho-baho (Lantana camara) aqueous extract using
the leaves, young stem, and flower in terms of the worms’ lethal time.
C. Procedure
The following Regulated Research institutions will be the venue of the
different tests to be conducted in the study:
Hicalde Farms in Sigma Capiz, it will be the site where we will collect the Eudrilus
euginiae and Roxas Memorial Provincial Hospital, it will be the site where the
experimentation will take place.
Plant Material and Extraction. Young stem, leaves and flower (~4-6 months
old) of Lantana camara will be obtained from Brgy. Cagay, Roxas City. Collected
samples will be air-dried for 4 days to retain <10% moisture and properly stored
in sealable plastic bags. A whole plant specimen will be subjected to
identification at the Department of Agriculture.
Air-dried Lantana camara will then be coarsely ground. Lantana camara will
be extracted using aqueous solution according to the recommendations of WHO
for medicinal plants based on the Philippine Pharmacopeia Annex A of A.O. No.
172, s.2004. This will ensure that antioxidant properties will be preserved as
they are more aqueous in nature. About 300g of each ground plant material will
be boiled in 1L distilled water in a beaker for 30 minutes at 100 oC. Then it will
then be cooled to room temperature and filtered through a filter paper.
Collection of earthworms. Seventy-two Eudrilus euginiae, nearly equal in
sizes (~6-8 cm) and weight (~1.5-2.5g), will be collected from the Hicalde Farms
in Sigma Capiz. A sample organism will be submitted for proper identification
at the Department of Agriculture.
Accumulation of earthworms. The collected Eudrilus euginiae will be
cleaned in natural saline solution to remove the soil and to prepare them for
the experimentation.
Preparation of the Different Treatments. Plant extract collected from the
stems will be diluted with normal saline to obtain 50% and 100%
concentrations. Albendazole will be used as the standard drug and will be
diluted with normal saline to obtain 1mg/ml concentration and poured into
Petri dishes. Normal saline (0.9% NaCl) alone will serve as the negative control.
Applying the Different Treatments. Eight groups of earthworms (n = 3/ petri
dish) will be utilized for the study. All treatment dilutions will be poured into
the Petri dishes accordingly containing 20 ml of each concentration.
Earthworms, nearly equal sizes (about 6-8 cm), will then be released in the
petri dishes containing each treatment at room temperature.
In vitro Anthelmintic Activity. The in vitro Anthelmintic Activity will be
performed according to the method of Ghosh et al. (2005) and Goswami et al.
(2011) with minor modifications.
A. Determining the time of paralysis:
Time for paralysis will be noted when no movement was observed with a
slight pin prick method by pricking the worms with a pin to check whether they
are paralyzed or dead. The time of paralysis will be recorded in terms of
minutes.
B. Determining the lethal time:
The lethal time will be recorded in terms of minutes after ascertaining that
the worms neither moved when shaken vigorously nor when dipped in warm
water (50°C).
Statistical Analyses. Data obtained will be further analyzed using Two-way
Analysis of Variance (ANOVA) to compare significant differences between
treatments and control. Statistical tool for Agricultural Research (STAR) 2.0.1
was employed. All inferential tests will be set at 5% level of significance.
References:

Ferreira, L. E., Castro, P. M. N., Chagas, A. C. S., França, S. C., & Beleboni, R.
O. (2013). In vitro anthelmintic activity of aqueous leaf extract of Annona
muricata L. (Annonaceae) against Haemonchus contortus from sheep.
Experimental Parasitology, 134(3), 327–332.
https://doi.org/10.1016/j.exppara.2013.03.032
Belizario, V. Y., Totañes, F. I. G., de Leon, W. U., Lumampao, Y. F., & Ciro, R.
N. T. (2011). Soil-transmitted helminth and other intestinal parasitic
infections among school children in indigenous people communities in
 Davao del Norte, Philippines. Acta Tropica, 120(SUPPL. 1).
https://doi.org/10.1016/j.actatropica.2011.02.010
Swathi, H., & Shekshavali, T. (2016). In Vitro Evaluation of Anthelmintic
activity of Rhus mysorensis leaves . Research Journal of Pharmacology
and Pharmacodynamics, 8(3), 115. https://doi.org/10.5958/2321-
5836.2016.00021.5
K, V. K., S, D. S., & Sukumar, E. (n.d.). Bio Chemistry Preliminary
Phytochemical Studies of Lantana Aculeata Roots, 365–368.
Kumar, K. V., Sharief, S. D., & Sukumar, E. (2011). Preliminary phytochemical
studies of Lantana aculeata roots. International Journal of Pharma and Bio
Sciences, 2(2), 365–368.
 Criteria in Evaluating A Research Plan YES NO
Title Is the title clear and unambiguous? ✓
Does it accurately reflect the content? ✓
Is it concise? ✓
Background, Have you demonstrated your awareness of relevant ✓
context literature, policy, theory or practice?
or rationale
Is the project relevant, important and timely? ✓
Aim and
objectives
Do the aim and objectives support the research question? ✓
Does the aim comprise a high-level statement about what
the researcher wishes to explore?
✓
Are the objectives specific and focused? ✓
If possible, are your objectives SMART? ✓
Hypothesis Is/are the hypothesis/es stated in the null form? ✓
Is it feasible to conduct the necessary statistical analysis
to test hypothesis the /es?
✓
Literature review Does it include a range of relevant literature (e.g.
scholarly, policy, in-house)?
✓
Does it present an appropriate selection of literature? ✓
Is the literature review well-structured and clearly written
(e.g. starting with a broad overview and then narrowing
✓
down to your topic)?
Research design
and methods
Are the research design and methods consistent with the
aim and objectives of your research?
✓
Is the methodology well described and used the
appropriate choice of method?
✓
Are the scope and boundaries of the project well defined? ✓
Are the number of subjects to be used in the study being
stated as well as how they will be selected?
✓
Ethical issues Are there ethical issues in this project? ✓
Risk assessment Are the risks identified in this project? ✓
Were the risk management given? ✓
Resources Are the materials readily available in the
environment/locality?
✓
Stated the facilities where the experiment will be
conducted?
✓

Julianne Pierrieta Lamazon

MODULE 1
 Included special facilities for working with biohazards or
other potentially dangerous substances.
✓
References Are all documents, websites and other resources fully and
accurately cited and referenced?
✓
Followed the preferred referencing style? ✓
Other Criteria Is the writing style clear, concise and to the point? ✓
Does the research plan have a logical flow? ✓
Used appropriate headings and other signposts to help the
reader?
✓
Absence of acronyms and jargon? ✓
Where there typographical and spelling errors? ✓
Are figures and illustrations used appropriately and
referred to in the text?
✓
Absence of non-essential text? ✓
A. From the title you selected in activity 1, make your research plan.
B. Evaluate the research plan you had made based on the criteria given above and
make the necessary corrections.
C. Submit your output to your teacher.

I Reflection
In this lesson, I learned that there are criteria for making a research plan and you
should follow these criteria.

I realized that it is important to check every detail of your research plan. It must be
accurate, concise, formal, etc.

I will apply the learnings I’ve got in the future and make our research plan better.