Chapter 1

Eleusine Indica Extracts As Antimicrobial

INTRODUCTION

As of today Microbes collectively, include bacteria, viruses, fungi, and parasites are
ramphant in our society that’s why for the past 70 years, antimicrobial drugs, such as
antibiotics, have been successfully used to treat patients with bacterial and infectious
diseases.However, mankind has been dependent on nature for their fundamental needs
since time immemorial and now one of the herbal plants used as antimicrobial is the
Paragis plant or Eleusine Indica, its scientific name . Today, there’s negligible studies
showing about the effect of Elusine Indica, however, it i s n o t ye t f u l l y p r o ve n .
P a r a g i s p l a n t ca n b e se e n a n ywh e r e b e c a u s e o f i t s h i g h adaptable to
different environment. Currently it is listed as invasive in several countries in Europe,
Asia, Central and South America and other countries. Elusine Indica is not yet
included on the approved list of herbal medicines of Department of Health, (Marvin
et.al, 2017). The researchers shed a light about the occurring presence and potential
of Paragis plant and to give answer to the questions and doubts about the rumors
on itspositive effect and also on global problem such as worldwide cases of
foodborne diseases.In most developing countries, the indigenous modes of herbal
treatment are a part of the culture and the dominant method of healing therapy. These
remedies are socially accepted because of their efficacy, safety, cost-effectiveness and
easy availability (Stuart et.al, 2017).

Medicinal plants have been used in traditional health care systems since prehistoric
times and are still the most important health care source for the most of the world’s
population (Bertrand Sagnia Sagnia (2014). Paragis went viral on social media sometime
in 2017. The weed once perceived an eyesore in anyone’s garden is claimed to have
curative powers. Despite its lack of official recognition from health authorities, peoples
from different cultures have been using paragis as a home remedy for certain ailments
according to Nila Eslit (2014). In Africa, Paragis is used as a natural way of treating
different kinds of illness. In Porac, Pampanga, the tribe of Aeta used it as an insect
repellent. The Aetas of Zambales are known to use Paragis to lower fever, against
inflammation and a natural antihistamine or anti-allergy treatment. In Surigao del Sur,
decoction of leaves drunk three time daily as diuretic. Paragis is declared to have protein
and its leaves have silicon monoxide, calcium oxide, and chlorine compounds. It is also
known to have numerous properties such as anti-inflammatory, antioxidant, antidiabetic,
antihistamine, natural diuretic, and cytotoxic properties. Its stem, roots, and leaves are
used as herbal medicine by boiling it. Paragis is an annual, erect, tufted, glabrous grass
usually eaten by Carabao & other farm animals in our country but it has actually 31
amazing health benefits in our body and one of it is the extract from leaves by decoction
that serves as antimicrobial.Before several studies was conducted, wire grass has been
used as folklore medicines by our ancestors even in other Asian Countries ( Den 2017).
 Plants produce a higher number of naturally occurring secondary metabolites, many
of them with unique pharmacologic activities. These metabolites include the flavonoids,
phenols and phenolic glycosides, saponins, cyanogenic glycosides, unsaturated lactones
and glucosinolates. In the past, herbs often represented the original sources of most
drugs and herbal remedies, but nowadays, alternative medicines are used widely in all
over the world. Herbal-derived remedies need a powerful and deep assessment of their
pharmacological qualities. With increasing recognition of herbal medicine as an
alternative form of health care, screening of medicinal plants for biologically active
compounds has become an important source of antibiotic prototypes and cancer-related
drugs. Hence, for selecting crude plant extracts with potential useful properties, in
vitro screening methods have been used for further in-depth chemical elucidation and
pharmacological investigations. To date, few studies of EI have been reported; specially,
its phytochemical content of sterol glucosides forms and C-glycosylflavone having anti-
inflammatory activity. To our knowledge, no other scientific investigation has been
reported in evaluating EI's therapeutic potential. Therefore, this study was conducted to
evaluate the antioxidant, antibacterial and anti-cancer activities of the hexane,
dichloromethane (DCM), ethyl acetate (EA) and methanol extracts (MeTH) of EI using
total phenolic content (TPC), DPPH, disc diffusion and MTT cytotoxicity assay methods
of analysis. Eleucine indica (EI) is traditionally used in ailments associated with liver and
kidneys. The therapeutic benefit of the medicinal plants is often attributed to their
antioxidant properties (Adel S. Al-Zubairi et.al, 2011).

This study can provide a wider knowledge about the Paragis plant and its health
benefits since people are dependent on nature by way of testing at the laboratory for
experimentation. Most of us usually preferred antimicrobial medicinal plants than other
antimicrobial drugs.
Conceptual Framework

The conceptual framework is developed after review of existing literature to investigate

the research questions at hand. This study will use Independent Variable and Dependent
Variable (IVDV) to show the cause-effect relationship of two variables as well as the
moderating variable. The Paragis leaf extract which is the independent variable is use to
determine the activity of the plant as antimicrobial which is the dependent variable. This
framework centers the inhibitory effect of Paragis leaf extract against different
microorganisms .The purpose of this is to identify and discover the health benefits of the
said plant.

Input Process Output

The inhibitory
Antimicrobial Activity effect of Paragis
 Paragis (Eleusine
of plant against
Indica) leaf
microorganisms
extract Paragis Plant

Figure 1. Schematic Diagram

 THE PROBLEM STATEMENT

Paragis plant is a common grass in our country and also to other countries outside the
Philippines. However, people did not make use further and proven its health benefits
specifically destroying microorganisms in our body. Medicinal plant cultivation could be a
prime occupation of people to upgrade their socio-economic status. The purpose of this
study is to determine the benefit or the effect of Eleusine Indica as antimicrobial and to
what extent it cures a certain illness.
Specifically, it seeks to asnwer the following questions:

1. What is the profile of the plant in terms of:

1.1 Color
1.2 Scientific Name
1.3 Structure

2. What are the characteristics of the plants?

3. How to cultivate and preserve Paragis Plant?

4. What are the substances found in Paragis leaf Extracts?

Significance of the Study

This study identifies the effectiveness of Paragis plant against

d i f f e r e n t microorganisms. The findings of th is stud y wo uld b e a great he lp
t o e ve ryone an d a re no te -wo rth y. Th is research would especially benefit the
following:

Medical practitioners. This study can be a ground for medical practitioners

especiallythe herbal users for using the Paragis plant for curing certain
diseases in terms of herbal medication or treatment.

Ethnobotanists. The study will help to extend the knowledge about the Paragis
plant favoring on the contribution of information of herbal plants.

Community. The utilization of Paragis plant for medication and treatment will be a
great benefit to the consumers especially those who prefer to use herbal medicines.
This study can be an orientation to the community in evidences and confirmations
about Paragis plant.
Scope and Delimitations

This study will focus on determining the inhibitory effect of Eleusine Indica extract
against pathogens related to foodborne illnesses. T h e researchers will give emphasis
on the procedures and methods bounded only with the study. The Paragis plant
and the isolation of different test microorganisms will be collected as part of
preparation of materials. Subsequently, the plant will be subjected toextraction through
decoction. The extract will be tested for its inhibitory effect against differenttest
microorganisms through agar-well diffusion method. This paper will only present
and discuss information about the antimicrobial screening of Paragis plant
against different microorganisms through textual and tabular presentation.

Definition and Terms

Antimicrobial – This refers destroying or inhibiting the growth of microorganisms and

especially pathogenic microorganisms.

Characteristics – This refers to a feature or quality belonging typically to a person, place,

or thing and serving to identify it.

Cultivation – It refers to the act or process of preparing the soil for the raising of crop.

Effect – a change that results when something is done or happens.

Eleusine indica – Scientific name of goosegrass or paragis plant.

Extract- it is a substance with necessary properties that is detached from the

plant typically by treating with a solvent and used in specific purpose such as testing.

Microorganism- it is a microscopic organism that can’t be seen by the naked eye.

Mostly, it causes different illnesses and weakens one’s body.

Paragis- a weed that usually grow tufted annual grass, prostrate and erect in v-shaped.

Socio-economic – This refers to relating to or concerned with the interaction of social

and economic factors.

S t r u c t u r e - T h i s re f e r s t o p a r t s t h a t co n st i t u t e s a s ys t e m .
 CHAPTER 2

REVIEW OF RELATED LITERATURE AND STUDIES

Ethnobotany

According to United States Department of Agriculture (USDA) 2016, ethnobotanyis the study
of a region's plants and their applied uses through the old-style knowledgeof a local
philosophy and individuals. In the beginning, ethnobotanical specimens andstudies were not
very reliable and sometimes not helpful. This is because the botanistsand the anthropologists did
not always collaborate in their work.

As detailed by Kochar (2016), ethnobotany means investigating plants in differentparts of the

world by original sources. In simply finding the middle grounds of definitionof ethnobotany, it
concludes about the critical study of plants and its application throughextensive aspects.

Pieroni (2016) stated that the botanists focused on identifying species and how theplants were
used instead of concentrating upon how plants fit into people's lives. On theother hand,
anthropologists were interested in the cultural role of plants and treatedo t h e r s c i e n t i f i c
a s p e c t s s u p e r f i c i a l l y. I n t h e e a r l y 2 0 t h c e n t u r y, b o t a n i s t s a n d anthropologists
better collaborated and the collection of reliable, detailed cross-disciplinary data
began.

Responses of a dinitroaniline-herbicide resistant (R) and a susceptible (S) goosegrass (Eleusine

indica) biotype to competition were determined in greenhouse studies by planting both biotypes
at various monoculture and mixture densities and proportions. In monoculture, reproductive
output per plant decreased more with increasing density in the R-biotype than in the S-biotype,
although no differences in above-ground biomass per plant were observed. Thus, the two
biotypes differed in their partitioning of resources to seed production. In mixture, the R-biotype
was more detrimentally affected by competition with the S-biotype than with itself. On the other
hand, the S-biotype was more affected by intra-biotype competition than by competition with
the R-biotype. Inter-biotype competition resulted in reduced tiller and inflorescence numbers,
above-ground biomass, and, especially, reproductive output in the R-biotype. Results indicated
that the R-biotype is less fit than the S-biotype.
MATERIALS AND METHODS
A resistant (Robeson) and a susceptible (Wake) goosegrass biotype were selected to study intra- and
inter-biotype competition. Plants were grown in the greenhouse as before (Chapter 2) in 15.2-cm
diameter pots. Experimental design was that of an addition series (23, 17) in randomized complete
blocks. Three experiments were conducted with 6, 7, and 5 replications, respectively. Densities used
were 0, 1, 4, 8, and 16 plants per pot (0, 55, 220, 440, and 880 plants/m2, respectively) of each biotype
including all possible combinations (maximum density 32 plants/pot or 1760 plants/m2). Treatment
arrangement provided a matrix of densities and proportions as depicted in Figure 2.1. Orientation of
the treatment matrix was randomized for each block. Since biotypes could not be differentiated
morphologically, resistant plants were identified with a plastic ring placed at the base of the stem.
Plants of each biotype were harvested and counted about 90 days after transplanting, when
inflorescences were mature but seed had not shattered. The following measurements were taken at
harvest: height, tiller number, leaf area, and inflorescence number. Harvested plants were oven dried
and dry vegetative, reproductive, and above-ground (vegetative plus reproductive) weights per
biotype were recorded. Data were analyzed by regression analysis, using appropriate
transformations, according to the methods suggested by Spitters (23,24). RESULTS AND DISCUSSION
Intra-biotype competition (monocultures). Plant size decreased with increasing density for both
biotypes. This result was expected because of the competition for limited resources among individual
plants. Reciprocal plant32 weight increased with plant density, i.e. plants became smaller as intra-
biotype competition became more severe. The R- and S-biotypes did not differ in reciprocal above-
ground weight per plant at any density. However, R- and S-biotypes allocated resources differentially
to vegetative and reproductive output under intra-biotype competitive conditions. The R- biotype
produced more vegetative weight than the S-biotype, differences being more marked as plant density
increased. On the other hand, the S-biotype outyielded the R-biotype in reproductive biomass with
increased intra-biotype competition. Since individuals of both biotypes were similar in maximum
vegetative, reproductive, and above-ground biomass, i.e. intercepts in regression model were
nonsignificant, this differential carbon allocation in favor of reproductive biomass in the S- biotype
suggests that the R-biotype is less fit than the S- biotype. In this regard, the dinitroaniline-resistant
biotype resembles triazine-resistant ones, which, with few exceptions (12, 21) are less competitive
than susceptible biotypes. The implications of such differential fitness have been pointed out by
Gressel and Segel (8). The selected biotype can continue to exist only where the selection pressure
(herbicide) is strong enough to keep the wild (S) type suppressed. Thus, reduced fitness is an
important factor in delaying the prevalence of herbicide resistance, and allowing the reestablishment
of S-populations of the weed species after herbicide use ceases. Reduced fitness in R-biotypes is most
important with less persistent herbicides. The change in leaf area with increasing density was best
described by a second-degree exponential polynomial . Both biotypes were similar in both the size of
the photosynthetic organs (intercept non significant) and in the rate of change with increasing
competition. Biotypes also were similar in height and tiller (total and fertile) number . The behavior
of the33 biotypes in monoculture indicated that both Robeson (R) and Wake (S) are similar in
vegetative characteristics but differ in carbon allocation patterns as density (competition) increases.
Inter-biotype competition (mixtures). Robeson (R) produced more vegetative weight than
Wake (S) in mixture (Figures 2.6-2.8, Appendix Table 2.2), but maximum attainable biomass was
similar for both biotypes (intercepts were not significant). The intra-biotype competition component
was more important for the S-biotype than for the resistant one. In mixture, the S-biotype had a more
detrimental effect on the R-biotype than vice versa. The S-biotype outyielded the R-biotype in
reproductive output. Intra-biotype competition resulted in greater reproductive weight decreases for
Wake (S) than for Robeson (R) . For the S-biotype, intra-biotype competition was more detrimental
than competition from the R-biotype. The opposite occurred with the R-biotype, with intra-biotype
competition having a lesser detrimental effect than competition with the S-biotype. The S-biotype
produced as much or more above-ground biomass than the R-biotype when growing in mixture. In
all experiments, intra-biotype competition effects on above-ground biomass were similar for both
biotypes. Similar to vegetative weight, inter-biotype competition was more detrimental for Robeson
(R) than for Wake (S). Similar trends were observed when leaf area per plant was evaluated. Robeson
(R) produced slightly more leaf area than Wake (S). The S-biotype also was more competitive than
the R-biotype with respect to both leaf area and tiller (total and fertile) production (Appendix Table
2.2). Our results emphasize the importance of dividing plant biomass into its main components when
harvesting competition experiments. If only above-34 ground biomass had been measured, we would
have concluded that no differential fitness between the biotypes existed in this study. The reduced
fitness of triazine-resistant biotypes is normally attributed to a less-efficient photosystem II, resulting
from the site of action modification responsible for triazine resistance. In the case of dinitroaniline
resistance, it is unlikely that a less-efficient photosynthetic apparatus is responsible for decreased
competitive ability, since the mechanism of action of these herbicides is not directly related to
photosynthesis. Dinitroanilines affect cell division, another physiological process of paramount
importance. If a modified tubulin molecule is responsible for dinitroaniline resistance as recent
research suggests, and if this modification impairs the ability of tubulin to polymerize into
microtubules or results in microtubules with a less-efficient physiological activity, it is not surprising
that R-goosegrass is less fit than S-goosegrass. Vaughn has indicated that a resistant biotype exhibits
malformed cell walls, which could result in impaired growth. However, we found that cell-wall
malformations are not common to all dinitroaniline-resistant biotypes, and, specifically, not present
in the R-biotype used in this study (Chapter 4).

LITERATURE CITED
1. APPLEBY, A. P., and B. E. VALVERDE. 1989. Behavior of dinitroaniline herbicides in plants. Weed
Tech. 3:(In Press).
2. APPLEBY, A. P., and C. STANGER. 1989. Italian ryegrass (Lolium multiflorum) accessions tolerant to
diclofop. Weed Sci. 37:(In press).
3. CONARD, S. G., and S. R. RADOSEVICH. 1979. Ecological fitness of Senecio vulgaris and Amaranthus
retroflexus biotypes susceptible or resistant to atrazine. J. Appl. Ecol. 16:171-177.35
4. CONNOLLY, J. 1986. On difficulties with replacement-series methodology in mixture experiments.
J. Appi. Ecol. 23:125-137.
5. DU PONT DE NEMOURS, E. I. and CO. 1988. Du Pont weed resistance workshop, Denver, CO.