Individual anti- typhoid activity of Ervatamia divaricata and

Imperata cylindrica as aqueous herbal syrups

Ida, A., Laureta, V.V., Aglugub, R., Bagalay, R., Dimaya, D.

Abstract

Typhoid fever is a bacterial disease caused by Salmonella typhi. Antibiotics are commonly used to treat
this disease, but resistance to these drugs are widespread. The aim of this study is to test the dose-response
relationships and efficacy of aqueous herbal syrups derived from individual plant extracts of Ervatamia
divaricata and Imperata cylindrica, administered orally in the treatment of typhoid fever in wistar rats that were
induced with S. typhi.

1. Introduction

Typhoid fever is an infection caused by Salmonella enterica, serovartyphibacteria, commonly referred to

as Salmonella typhi. Salmonella paratyphi causes a less severe febrile illness, and other species of Salmonella
bacteria cause other, usually less severe infections. The annual incidence of typhoid is estimated to be about 17
million cases worldwide, and is highest in those between the ages of 5 and 12 years. In Southeast Asia, the
incidence of typhoid fever varies widely between sentinel sites (annual incidence: 24/100 000 person years in
Vietnam, 180/100 000 person years in Indonesia, 494/100 000 person years in India).

Approximately 420 000 deaths occur annually in Asia due to typhoid fever. Without treatment, case-
fatality rates of infection are 10%. With appropriate antibiotic therapy, case-fatality rates can be reduced to
below 1%. Between 1 January and 13 November 2013, 28 224 cases of suspected or clinically diagnosed typhoid
fever were recorded in the Philippines. Two of these cases resulted in death, yielding a case-fatality rate of
0.27%

During the same time period in Regions 6, 7, and 8 and the National Capital Region, there were 5 637
suspected or clinically diagnosed cases and 60 laboratory-confirmed cases Although the infection is treatable
with antibiotics, treatment is complicated by growing resistance to widely available oral antibiotics in several
areas of the world, including Southeast Asia.

People can transmit the disease as long as the bacteria remain in their system; most people are
infectious prior to and during the first week of convalescence. About 10% of untreated patients will discharge
bacteria for up to three months; 2 to 5% of untreated patients will become permanent carriers.

Salmonella typhi is passed in the feces or urine of infected people. People become infected by ingesting
food or drink that has either been handled by an infected person or contaminated by sewage containing the
bacteria. In less industrialized countries, transmission commonly occurs where there is inadequate sewage
disposal and flooding, or otherwise unsafe drinking water. Where water quality is high, transmission is more
likely to occur via food contaminated by healthy carriers of Salmonella typhi.

Typhoid fever typically presents as a febrile illness that affects infants, young children and adults. The
onset of symptoms takes place 1-3 weeks after exposure. Patients should be suspected of typhoid fever if they
present with persistent high fever (>7 days), abdominal manifestations (abdominal pain, vomiting, bloating,
constipation, soft stools) and any of the following: weakness, poor appetite, enlarged liver and spleen, rose
spots on the chest/abdomen, and a relatively slow heart rate. Older children and adults may experience
constipation, while younger children may have diarrhea.

Severe forms of disease may involve delirium, shock, and intestinal perforation or hemorrhage.
Immunocompromised people are at increased risk for severe disease.

Tabernaemontana pandacaqui Poir., (synonym: Ervatamia pandacaqui (Poir.) Pichon) (Apocynaceae) is

known in Thailand as ‘Phut’. The root of plant is boiled with water and used in folklore medicine for the
treatment of pain and inflammation (Phu-pattanapong, 1979). Tabernaemontana plants also are the source of
several indole alkaloids (van Beek et al., 1984).

Antimicrobial drugs either kill microbes (microbicidal) or prevent the growth of microbes (microbistatic).
The plant named Imperata cylindrica is commonly known as lalang and is traditionally used by the Malay
community in folk remedies for cancer, colds, diarrhea, dysentery, gonorrhea, myalgia, night sweats, piles,
rheumatism, and tumors. Cogongrass or Imperata cylindrica is a perrenial, rhizomatous grass that is somewhat
variable in appearance. Young leaves are light green while older leaves are orange- brown to brown in color.
The dead leaves remain standing and resist decay. Each individual flower spikelet has two stamens and two
feathery stigmas and is attached to a fuzzy plume that later assists the wind-dispersed seed in drifting through
the air. In temperate areas Imperata cylindrica usually flowers from late winter through May or in the fall after
the first frost. It may flower year-round in more tropical areas. Stands of this species are sometimes burned or
cut so that the tender new growth can be used for short term supplemental or emergency pasture, but it
generally produces poor quality forage and animals avoid chewing the sharp- edged mature leaves. Imperata
cylindrica is often planted for soil stabilization and sometimes used for roofing thatch. So, this little known plant
had not screened scientifically.

T. pandacaqui is proven to possess analgesic and antipyretic effects (Taesotikul et al., 1989). While I.
cylindrical is proven to be antibacterial.

In this study, individual aqueous extracts are derived from these two plants. An individual aqueous
extract will be derived from the dried stems of T. pandacaqui and same individual extract will be derived from
the aerial parts of I. cylindrica. This aqueous extracts will be individually assayed for purity and content. Then
after, these two extracts will be formulated individually to a standard aqueous herbal syrup. The aqueous herbal
syrups will be assayed for content and purity using pharmacopeia-derived standardization methods.

Healthy rats will be induced with S. typhi strain and will be orally given a dose of these formulations. To
test the efficacy of these two formulations in treating typhoid fever, dose- response relationships method will be
used.

2. Materials and methods

2.1 Bacterial strain

The bacterial strain is a pure culture of S. typhi. The bacteria is previously cultured and isolated in a
microbiology laboratory.
2.2 Animals

Healthy Wistar rats will be used. Ethics for the use of the animals will be secured beforehand before
conducting the study. The animals 6-9 weeks of age, will be housed individually in macrolon cages. The animals
will be acclimatized for at least 1 week before beginning the experiment. The breeding colony of the animals
will be pre- screened and monitored for endogenous pathogenic viruses and bacteria. The rats will be induced
with typhoid fever by injecting a significant dose from an isolated pure culture of S. typhi.

2.3 Plant and material extraction.

Stems of T. pandacaqui and aerial parts of I. cylindrica will be collected from the locality. The two plants
will be individually made into a standard aqueous extract and assayed for purity and content. After
which, they will be individually formulated to a standard aqueous herbal syrup and assayed again for
conformity to standards.

2.4 Anti-typhoid activity testing

After injecting the animals with the bacterial strain, the animals will be left for 6 days. Widal test will be
used to test the presence of typhoid fever disease in the animals. Widal Test is an agglutination test which
detects the presence of serum agglutinins (H and O) in patients serum with typhoid and paratyphoid fever.
Then right after confirmation of the disease, the animals will be given therapeutic doses of the two
formulations. Data will be recorded.

2.5 Dose- response relationship method

Data will be gathered and statistical methods will be employed to check the dose- response relationships
present in this study.

3. Results

As soon as the data are available, results can be delivered.

4. Discussion

5. Conclusion
Review of related literatures:

Immunization, Vaccines and Biologicals

Typhoid
Typhoid fever is a systemic infection caused by Salmonella Typhi, usually through ingestion of contaminated
food or water. The acute illness is characterized by prolonged fever, headache, nausea, loss of appetite, and
constipation or sometimes diarrhoea. Symptoms are often non-specific and clinically non-distinguishable from
other febrile illnesses. However, clinical severity varies and severe cases may lead to serious complications or
even death. It occurs predominantly in association with poor sanitation and lack of clean drinking water.
According to the most recent estimates (published in 2014), approximately 21 million cases and 222 000
typhoid-related deaths occur annually worldwide. A similar but often less severe disease, paratyphoid fever, is
caused by SalmonellaParatyphi A, B or C.

Two typhoid vaccines are currently recommended for use by:

 an injectable polysaccharide vaccine based on the purified Vi antigen (known as Vi-PS vaccine)
for persons aged two years and above;
 and a live attenuated oral Ty21a vaccine in capsule formulation for those over five years of age.
WHO recommends the use of the Vi-PS and Ty21a vaccines to control endemic disease and for outbreak control.
WHO further recommends that all typhoid fever vaccination programmes should be implemented in the context
of other efforts to control the disease, including health education, water quality and sanitation improvements,
and training of health professionals in diagnosis and treatment.

Several Vi polysaccharide–protein conjugate vaccine candidates are under development (or are nationally
licensed but not on the international market) and anticipated to be available in the future for infant
immunization.

http://www.who.int/immunization/diseases/typhoid/en
 Anti-inflammatory, antipyretic and antinociceptive activities of
Tabernaemontana pandacaqui Poir
T. Taesotikul a, A. Panthong a,*, D. Kanjanapothi a, R. Verpoorte b, J.J.C. Scheffer b
a
Department of Pharmacology, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand
b
Division of Pharmacognosy, Leiden/Amsterdam Center for Drug Research, Leiden University, 2300 RA Leiden, The Netherlands

Received 4 January 2002; received in revised form 3 September 2002; accepted 3 September 2002

Abstract

Studies on carrageenin-induced rat paw edema, yeast-induced hyperthermia in rat and writhing response induced by acetic
acid in mice showed that the alcoholic extract of stems of Tabernaemontana pandacaqui (T. pandacaqui ) has significant anti-
inflammatory, antipyretic and antinociceptive activities. These activities are due to alkaloidal components since they were a lso
observed when the crude alkaloidal (CA) fraction separated from alcoholic extract was tested in the same models.

# 2002 Elsevier Science Ireland Ltd. All rights reserved.

Keywords: Tabernaemontana pandacaqui ; Alcoholic extract; Crude alkaloidal fraction; Anti-inflammatory; Antipyretic; Antinociceptive

https://www.researchgate.net/publication/10976342

Antibacterial Activity of Aerial Parts of Imperata cylindrica (L) Beauv.

V. Parkavi, M. Vignesh*, K. Selvakumar, J. Muthu Mohamed, J. Joysa Ruby

Faculty of Pharmacy, Masterskill University College of Health Sciences, Batu 9, 43200 Cheras, Selangor, Malaysia

ABSTRACT
Antimicrobial is an agent that kills or inhibits the growth of microorganisms such as bacteria, fungi, or protozoan,
as well as destroying viruses. Antibacterial drugs either kill bacteria (bactericidal) or prevent the growth of bacteria
(bacteriostatic). The scope of this study was to extract the active constituents present in the plant Imperata
cylindrica by cold maceration method and to find the folklore claim of antibacterial action. The antibacterial activity
was performed under sterile condition by using cup and plate method. Three extracts were prepared for this study
such as ether extract, ethanolic extract and aqueous extract. The extractions were diluted as 200 mg/2 ml ratio.
Each 200 mg of aqueous extract and ethanol extract was diluted with 2 ml of distilled water respectively; whereas
the ether extract was suspended with tween 80, a suspending agent. The antibacterial test was tested against
Staphylococcus aureus and E. coli. The test was done in triplicate with sterile Petri plates (10×10 cm) in Muller
Hinton agar media. 0.5 ml of diluted culture was poured on each Petri plates and a well of 8 mm diameter
approximately was cut with sterile metallic borer in the inoculated agar plate. The wells were filled with previously
diluted three different extracts separately. The plates were labeled and incubated for 24 hours at 37°C. At the end
of incubation period, the zone of inhibition (diameter) was measured and results showed that aqueous extract had
very potent antibacterial activity comparatively with other extracts.

Keywords: Antibacterial, Imperata cylindrica, E. coli, Staphylococcus aureus.

www.ijpsdr.com
References:

World Health Organization. (n.d.). Immunization, Vaccines and Biologicals. World Health Organization. Retrieved
March 16, 2018, from http://www.who.int/immunization/diseases/typhoid/en

T. Taesotikul a, A. Panthong a,*, D. Kanjanapothi a, R. Verpoorte b, J.J.C. Scheffer b. (2002). Anti-inflammatory,

antipyretic and antinociceptive activities of Tabernaemontana pandacaqui Poir. 2002 Elsevier Science Ireland All
rights reserved. Retrieved March 16, 2018, from https://www.researchgate.net/publication/10976342.

V. Parkavi, M. Vignesh*, K. Selvakumar, J. Muthu Mohamed, J. Joysa Ruby. (2012). Antibacterial Activity of Aerial
Parts of Imperata cylindrica (L) Beauv. International Journal of Pharmaceutical Sciences and Drug Research.
Retrieved March 16, 2018, from www.ijpsdr.com.
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