MAHARASHTRA UNIVERSITY OF HEALTH SCIENCE

A SYNOPSIS FOR DISSERTATION FOR

M.D. (AYURVEDA) IN THE SUBJECT OF ROGNIDAN AVUM VIKRITIVIGYAN

TITLE:

In vitro evaluation of antibacterial effect of Dronapushpi patra swaras (Leucas cephalotes

spreng) on Pseudomonas aeruginosa compare with ciprofloxacin as a control drug by culture
and sensitivity method

NAME OF THE CANDIDATE

DR. KUNDAN BHARAT LADE
P.G. SCHOLAR 1ST YEAR

(DEPARTMENT OF ROGNIDAN AVUM VIKRITIVIGYAN)

GUIDE

DR. SATYENDRAKUMAR SINGH

ASSO. PROFESSOR
(ROGNIDAN AVUM VIKRITIVIGYAN)

H.O.D.
DR. RAGINI R. PATIL
PRINCIPAL, PROFESSOR & HOD
(DEPT. OF ROGNIDAN AVUM VIKRITIVIGYAN)

NAME OF THE INSTITUTE

PUNE DISTRICT EDUCATION ASSOCIATION'S
COLLEGE OF AYURVEDA AND RESEARCH CENTER NIGDI PUNE
YEAR 2021-2022
 MAHARASHTRA UNIVERSITY OF HEALTH SCIENCE, NASHIK.

(For submission of proposal of the university)

1. Academic 2021-2022
2. Subject Rognidan Avum Vikruti Vigyan
3. Name Of Student Dr. Kundan Bharat Lade
4. Name Of Guide and Designation Dr. Satyendrakumar Singh
Associate Professor
5. Eligibility Yes
6. Recognition of Guide MUHS MUHS/E-
Letter No. 3/PG/122105/31/01/2615/2021

7. Guide having approved 05 year Yes

Teaching
PL mention total approved Teaching
Experience

8. FEES:
DD NO.
Date
9. Appendix A (Format for submission Yes
Of topic by P.G. STUDENT)
10. Appendix B (Report of Ethics Committee) Yes

Seal of college Signature

Dean/Principal

2
 APPLICATION BY THE CANDIDATE FOR SUBMITTING THE SYNOPSIS

Dr.Kundan Bharat Lade

Pune District Education Association’s
College of Ayurveda and Research Centre,
Nigdi, Pune 411044

To.
The Registrar,
Maharashtra University of Health Science,
Nashik

Respected sir /Madam,

Submitting here with the synopsis for M.D. Ayurveda for your kind of observation
and approval.

Thanking you in anticipation

Dr. Kundan Bharat Lade

P.G. Scholar 1st year

Forwarded Through

GUIDE HOD, PRINCIPAL

Dr. Satyendrakumar Singh Dr.Ragini R. Patil
M.D.(Rognidan Avum Vikriti Vigyan) M.D.(Rognidan Avum Vikriti Vigyan)

3
 A’ FORMAT FOR SUBMISSION OF TOPIC BY P.G. STUDENT)

Name of P.G. College Pune District Education Association’s College of Ayurveda

and Research Centre, Nigdi, Pune 411044

Department Rognidan Avum Vikriti Vigyan

Name of Guide and College Dr. Satyendrakumar Singh

Associate Professor
M.D.(Rognidan Avum Vikriti vigyan)
Pune District Education Associations College of Ayurveda
and Research Centre, Nigdi, Pune 411044
Contact No. 9890741683

To,
The Registrar,
MUHS, Nashik-422004
Subject: Submission of Title of and synopsis of Dissertation
Respected Sir/Madam,
I, Kundan Bharat Lade registered for M.D. (Ayurveda) in Rognidan Avum Vikriti Vigyan 2021-
2022 batch under the guidance of Dr. Satyendrakumar Singh, Pune District Education
Association College of Ayurveda and Research Centre, Nigdi, Pune.
I am due to appear for Rognidan Avum Vikriti vigyan in 2022. I am submitting here with Title
and Synopsis of Dissertation as mentioned below and as suggested by my aforesaid guide,
“In vitro evaluation of antibacterial effect of Dronapushpi patra swaras (Leucas cephalotes
spreng) on Pseudomonas aeruginosa compare with ciprofloxacin as a control drug by culture
and sensitivity method”.

Dr. Satyendrakumar Singh Dr.Ragini R. Patil Dr. Kundan Bharat Lade

(GUIDE) (DEAN, HOD & PRINCIPAL) (CANDIDATE SIGN)

4
 Sr. ITEM Components
No
1. Title In vitro evaluation of antibacterial effect of Dronapushpi patra swaras
(Leucas cephalotes spreng) on Pseudomonas aeruginosa compare with
ciprofloxacin as a control drug by culture and sensitivity method
2. Introduction Ayurveda deals with good, bad, blissful and sorrowful life, and with what is
wholesome and unwholesome for it, longevity, and about what Ayu is in
itself. It implies the conjunction of mind and soul.There are so many reasons
to cause harmful effects on human body. It includes mental stress, change
in lifestyle, and lack of exercise, bad eating habits, trauma, and pathogenic
microorganism like bacteria, virus, fungi and protozoans1.
In ayurved samhitas description of krimi is found, krimi word is used as broad
sense for all worms and microorganisms. In modern medical science there is
separate branch for worms and microbes that is helminthology and
microbiology.

वििंशविविध ाः विमयाः पूिवमुविष्ट न न विधेन प्रविभ गेन न्यत्र सहजेभ्याः; िे पुनाः

प्रकृविवभविवभज्यम न श्चिुविवध भिन्ति; िद्यथ - पुरीषज ाः, श्लेष्मज ाः, शोवििज , मलज श्चेवि||
च. वि. ७/९

In Charak samhita twenty types of pathogenic parasites have been

mentioned. They, being divided as per their source of genesis and are of four
types, developing in feces, in mucous, in blood and developing from external
excreta2.
सुरस श्वेिसुरस फविज्झक जवकभूस्तृिसुगन्धकसुमुखक लम लकुठे रकक समर्व क्षिकखरपु
ष्प विडङ्गकट् फलसुरसीवनगुवण्डीकुल हलोन्र्ु रुकविवक फञ्जीप्र चीबलक कम च्यो विषमुवष्टक
श्चेवि|| सु. सू. ३८/ १८
सुरस वर्गविो ह्येष कफहृि् कृवमसूर्नाः | प्रविश्य य रुवचश्व सक सघ्नो व्रिशोधनाः|| सु.सू.३८/
१९
As described in Sushrut samhita, Dronapushpi (Leucas cephalotes spreng.) is
included in Surasadi gana and it has krimighna property. Leucas cephalotes
spreng (Dronapushpi) a medicinal plant belonging to family Lamiaceae,
found as a common weed throughout the India. This is found in rainy season

5
and whole plant is used for medicine. The leaves of Dronapushpi are used in
fevers and urinary tract infections. Similarly in modern medicine
fluoroquinolones are used in urinary tract infection. In fluoroquinolones
ciprofloxacin has good efficacy against gram negative bacteria3-6.
The Pseudomonas aeruginosa is a gram negative bacteria, motile, non-
capsulated that grows well on ordinary media producing large, opaque,
irregular colonies with distinctive musty earthy smell.It can be found in
water, soil, plants, insects and other animals including humans.
Pseudomonas aeruginosa can lead to wide range of disease states like
nosocomial infection, urinary tract infection, sepsis, diarrhea, soft tissue
infection. Pseudomonas aeruginosa is a frequent cause of urinary infection
following catheterization, the role of Pseudomonas aeruginosa in hospital infection
is due to its resistance to common antibiotics. It grows best between 350c to
370c. Pathogenicity means it’s an ability of microbe to cause disease.
Bacterial culture mean bacteria have to be grown on culture media and
obtained as pure cultures for study7-11.
Urinary tract infection is most common bacterialinfection. Especially women
are more likely to get infected. The prevalence of urinary tract infection in
women is 3% at the age of 20 year.The infection is mostly caused by gram
negative bacteria such as Pseudomonas aeruginosa, E. coli. Sometimes it
becomes impossible to eradicate UTI because of development of drug
resistant bacteria. Hence prior isolation of causative organisms and their
sensitivity to antimicrobial drugs should be done before any rational
treatmentis given to the patient12.
In Culture and Sensitivity, for culture midstream urine is collected
aseptically in sterilized container. Bacterial counts on fresh, voided urine
from infected patient show more than 100,000 (105) organism per millilitre
significant bacteriuria. However, bacterial counts of less than 105 per
milliliter may occur in patient on antibacterial therapy or in patients who
are excessively hydrated causing dilution of urine. Urine is an excellent
culture medium for bacteria. For isolation and identification of

6
 Pseudomonas aeruginosa cetrimide agar medium is used13-15.
Rationale of the study: -
Acharya Sushrut have mentioned Dronanpushpi which have krimighna
property. Pseudomonas aeruginosa is one of the causative organism for
urinary tract infection. So if Dronapushpi found effective against
Pseudomonas aeruginosa it will help in management of urinary tract
infection and it may be an alternate to the resistant antibiotics. Hence this
topic is selected.
3. Primary Whether the Dronapushpi patra swaras (Leucas cephalotes spreng) is as
Research effective as ciprofloxacin against Pseudomonas aeruginosa?
Question
4. Primary Null Hypothesis (H0) - Dronapushpi patra swaras is not as effective as
Hypothesis ciprofloxacin against Pseudomonas aeruginosa.
Alternate Hypothesis (H1) - Dronapushpi patra swaras is as effective as
ciprofloxacin against Pseudomonas aeruginosa.

5. Review of 1. The literary review regarding Krimi and Dronapushpi will be studied
literature from various ayurveda text and samhitas.
2. Modern review related to culture and sensitivity and Pseudomonas
aeruginosa will be taken from textbook of microbiology by Anantnarayan
and Paniker, The short textbook of medical microbiology by Satish gupte.
Similarly modern review related to ciprofloxacinwill be studied from K.D
Tripathi essentials of medical pharmacology.
3. Internet data and various research articles from Google Scholar,
Shodhganga, DHARA, and Pubmed related with Pseudomonas
aeruginosa, antibacterial effect of Dronapushpi (Leucas cephalotes
spreng) and ciprofloxacin and culture and sensitivity will be studied.
Previous workdone: -
1. Deogade MS, Kethamakka SRP. Krumi (Microorganisms) in
Ayurveda- A critical review. International Journal of Ayurvedic
Medicine[Internet].2020;10(4):297300.Availablefrom:http://dx.d

7
 oi.org/10.47552/ijam.v10i4.1326
2. Bekele T, Tesfaye A, Sewunet T, Waktola HD. Pseudomonas
aeruginosa isolates and their antimicrobial susceptibility pattern
among catheterized patients at Jimma University Teaching
Hospital, Jimma, Ethiopia. BMC Res Notes. 2015 Sep 28;8:488.
doi: 10.1186/s13104-015-1497-x. PMID: 26416559; PMCID:
PMC4587897.
3. Itoria Priyank and Jain Gaurav – To investigate Leucas cephalotes
for its antioxidant and hepatoprotective activity against cc14
induced hepatotoxicityin swiss albino mice, international journal
of pharma and bio sciences, vol 3 /issue 2 / 2012.
4. Rohini Shinde B*., Vaibhav Jagtap A., Umesh Joshi P., R.B. Patil and
R.Dwagh - on in vitro antihelminthic activity of Leucas cephalotes
leaf extract, international journal of drug discovery and herbal
research 2011.
5. Katura Antariksh, Pradhan Chandan Kumar, Tyagi Amit Kumar,
Singh Pradeep – Antibacterial activity of Leucas cephalotes Roth
spreng whole herb.2010.
Present Knowledge gap: -
In this previous works, there are many works done related to
antidiabetic property, hepatoprotective property, anti-inflammatory
property, analgesic property of dronapushpi using different parts but till
now no work has been done on In vitro evaluation of antibacterial effect
of Dronapushpi patra swaras (Leucas cephalotes spreng) on
Pseudomonas aeruginosa compare with ciprofloxacin as a control drug
by culture and sensitivity method

6. Primary To study in vitro evaluation of antibacterial effect of Dronapushpi patra

Objective swaras (Leucas cephalotes spreng) on Pseudomonas aeruginosa compare
with ciprofloxacin as a control drug by culture and sensitivity method.

8
7. Methodology I. Appropriate Study Design -
It is an experimental analytical study.
II. Mentioned Study setting-
The proposed study will be done in 3 levels.
Level 1 (Conceptual) - Ayurvedic literature regarding dronapushpi patra and
its antibacterial property similarly modern concept regarding Pseudomonas
aeruginosa, culture and sensitivity and ciprofloxacin will be studied.

Level 2 (Experimental) – Observation of the bacterial growth and sensitivity

by culture and sensitivity method will be done.

Level 3 (Analytical) – Antibacterial effect of dronapushpi patra swaras and

ciprofloxacin on pseudomonas aeruginosa in the form of zone of inhibition in
mm will be noted.

Study Duration- 18 months

Study Place -
Institutional Pathology Laboratory.

III. Mentioned Study Population-

The standard culture of microorganism Pseudomonas aeruginosa.

IV. Sample size- Not applicable.

V. Appropriate sampling technique- Not applicable.

VI. Method of Selection of study subject-

Appropriate Inclusion criteria –
Standard culture of microorganism Pseudomonas aeruginosa will be
taken.

9
 Appropriate Exclusion criteria – Not applicable
Appropriate subject withdrawal criteria - Not applicable
VII. Operational definitions provided- Not applicable
VIII. Appropriate methods of measurements-
Objective Criteria-
Table no.1 – Dronapushpi patra swaras and ciprofloxacin distribution for
Pseudomonas aeruginosa according to Sensitivity and resistance.

Dronapushpi Patra Ciprofloxacin

Swaras
Sensitive culture
Resistant culture
Total

Table no 2–Zone of inhibition and percentage wise distribution for

Pseudomonas aeruginosa

Zone of Dronapushpi % Ciprofloxacin %

inhibition patra swaras
in mm
0 to 4
4 to 12
> 12
Total

IX. Appropriate Study instrument / data collection tools-

Dronapushpi patra swaras disc, ciprofloxacin drug disc, standard culture of
Pseudomonas aeruginosa, petri dish, incubator, inoculation loop, gloves.

X. Methods of data collection relevant to objective-

Identification of Pseudomonas aeruginosa –
Colonies of Pseudomonas aeruginosa appears green in color, large, smooth,
flat and elevated edges. And shape of colonies is small round.
-Microscopic examination, gram staining and chemical test will be done for

10
 identification of Pseudomonas aeruginosa
-Indole, Methyl Red, Voges Proskauer (VP), H2S, Catalase, Oxidase and
arginine dihydrolase test will be done.
Dronapushpi Patra Swaras Preparation -
1) 5 gm of fresh leaves of dronapushpi of properly grown dronapushpi plant
will be taken.
2) It will be washed with water by making it sterile, the leaves will be
smashed and grind.
3) Smashed material will be squeezed with sterile cotton cloth and swaras
will be collected in sterile measuring cylinder.
4) Swaras will be autoclaved.

Preparation of dronapushpi patra swaras disc –

1) Paper disc will be prepared from whartman filter paper with the help of
paper punching machine.
2) 5 microlitre swaras will be taken in micropipette and added on paper
disc.
3) Disc will be placed in sterile petri dish and allow to dry.
4) Disc will be dried for 45 min.
Disc will be autoclaved.

During this research study the material and methodology will be asfollows:
Literary Review

Dronapushpi patra swaras will be prepared.

From swaras, disc will be prepared.

Standard culture plate of Pseudomonas aeruginosa will be taken.

11
Drug disc of Dronapushpi patra swaras and ciprofloxacin will be placed on
standard culture plate of Pseudomonas aeruginosa.

Standard culture plate will be placed in incubator at 370c for 24 hours

After 24-hour Sensitivity of Dronapushpi patra swaras and .ciprofloxacin will

be observed

For determination of Sensitivity, the zone of inhibition will be measured in

mm.

The mean of zone of inhibition of Dronapushpi patra swaras and

Ciprofloxacin will be calculated.

Result will be drawn.

XI. Appropriate Data Management and analysis procedure-

Observations will be noted in the form of chart, graphs and
tables.

XII. Appropriate data analysis plan and method-

Mean and percentage of zone of inhibition will be calculated and
result will be noted.

XIII. Additional points for research in ayush-

Not applicable.

XIV. Additional Points For RCT-

Not applicable

XV. Additional points for all experimental study-

Not applicable

12
8. Reference 1. SamhitaC.Deerghanjiviteeyaadhyaya[Internet].CharakSamhita.2022[
cited2022Aug4].Availablefrom:https://www.carakasamhitaonline.co
m/index.php?title=Deerghanjiviteeya_Adhyaya&oldid=41131
2. Samhita C. Janapadodhvansaniya vimana [Internet]. Charak Samhita.
2022[cited2022Aug4].Availablefrom:https://www.carakasamhitaonli
ne.com/index.php?title=Janapadodhvansaniya_Vimana&oldid=4112
3. E-Samhita - national institute of Indian medical heritage [Internet].
Nic.in.[cited2022Aug6].Availablefrom:https://niimh.nic.in/ebooks/es
ushruta/?mod=read
4. Chauhan M. Dronapushpi (Leucas cephalotes) - Properties, Benefits &
Dosage [Internet]. Planet Ayurveda. 2019 [cited 2022 Oct 4]. Available
from: https://www.planetayurveda.com/library/dronapushpi-leucas-
cephalotes/
5. Madhukiran B.L., Vijaya Lakshmi K, and Uma Maheswhari Devi P,on-
antibacterial properties of Leucas cephalotes (roth) spreng , Leaf,
ancient science of life , vol: xxi (4) 2002
6. Bekele T, Tesfaye A, Sewunet T, Waktola HD. Pseudomonas
aeruginosa isolates and their antimicrobial susceptibility pattern
among catheterized patients at Jimma University Teaching Hospital,
Jimma, Ethiopia. BMC Res Notes. 2015 Sep 28;8:488. doi:
10.1186/s13104-015-1497-x. PMID: 26416559; PMCID: PMC4587897
7. Ananthanarayan R, Jayaram Paniker CK. Ananthanarayan and
paniker’s textbook of microbiology. 7th ed. Universities Press; 2010.
Chapter No.30 Enterobacteriaceae 1: Coliforms-Proteus Page No. 280
8. Gupte S.The short textbook of medical microbiology (Including
Parasitology).New Delhi,India:Jaypee Brothers Medical;2010 Chapter
No. 31 Pseudomonas Page No. 217
9. Ananthanarayan R, Jayaram Paniker CK. Ananthanarayan and
paniker’s textbook of microbiology. 7th ed. Universities Press; 2010.
Chapter no. 30, Page No. 281

13
 10. Gupte S.The short textbook of medical microbiology (Including
Parasitology).New Delhi,India:Jaypee Brothers Medical;2010 chapter
no.9 infection page no. 58
11. Anantnarayan R, Jayaram Paniker CK.Anantnarayan and panikers
textbook of microbiology.8th ed. Universities Press; 2010. Chapter no
4, Page no.39
12. Penman ID, Ralston SH, Strachan MWJ, Hobson R, editors. Davidson’s
principles and practice of medicine. 23rd ed. London, England: Elsevier
Health Sciences; 2018 chapter no.15 nephrology and Urology Page
no.426
13. Gupte S. The short textbook of medical microbiology (Including
Parasitology). New Delhi, India: Jaypee Brothers Medical; 2010.
Chapter No.63 Diagnostic Microbiology - An Approach to Laboratory
Diagnosis Page no.418
14. Penman ID, Ralston SH, Strachan MWJ, Hobson R, editors. Davidson’s
principles and practice of medicine. 23rd ed. London, England: Elsevier
Health Sciences; 2018 chapter no.15 nephrology and Urology Page
no.427
15. Aryal S. Cetrimide Agar- Composition, Principle, Uses,Preparation
andColonyMorphology[Internet].MicrobiologyInfo.com.Microbiolog
yInfo;2015[cited2022Oct4].Availablefrom:https://microbiologyinfo.c
om/cetrimide-agar-composition-principle-uses-preparation-and-
colony-morphology/

9. Annexure Timeline/Gant chart

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 Annexure
Timeline / Gantt chart
Name –
Department –
Starting date –
Date of approval of synopsis-

Months 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18

Activities

Review of Y Y Y Y
literature

Visit to center Y Y Y Y Y Y

Data entry as Y Y Y Y
a master chart

Dissertation Y Y Y Y Y Y
writing

Dissertation Y
binding

Signature of Y
concern
authority

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