A

PROJECT REPORT
IN THE SUBJECT OF MICROBIOLOGY

“ANTIMICROBIAL ACTIVITY OF LEAF EXTRACT OF

PROSOPIS CINERARIA”
SUBMITTED TO THE
SHRI JAGDISH PRASAD JHABARMAL TIBREWALA UNIVERSITY
FOR THE DEGREE OF
B.SC.
Submitted to
the

In Partial Fulfilment of

B.Sc. (Bioscience) VI Semester

BY
JYOTI SWAMI
(Roll No.20BG1003)
UNDER THE GUIDANCE OF

DR.ARUN KUMAR

SHRI JAGDISH PRASAD JHABARMAL TIBREWALA UNIVERSITY,

VIDYANAGARI, JHUNJUNU, RAJASTHAN -33001

YEAR, 2022-23
 DECLARATION BY THE CANDIDATE

I assure that the research entitled, "ANTIMICROBIAL ACTIVITY OF LEAF EXTRACT

OF PROSOPIS CINERARIA" supervised by DR.ARUN KUMAR Associate
Professor at Shri Jagdish Prasad Jhabarmal Tibrewala University, Vidhyanagari
in Rajasthan, and accepted by the Research Degree Committee. My work is my
own I've worked with the supervisor.

Any work submitted for credit at this, or any other institution/deemed

universityhas not been revised without my knowledge, I guarantee you.

Signature of Supervisor Signature of the Candidate

 CERTIFICATE OF THE SUPERVISOR

This is to assure that work entitled, " ANTIMICROBIAL ACTIVITY OF LEAF EXTRACT OF
PROSOPIS CINERARIA " is a piece ofresearch work done by JYOTI SWAMI, and submitted
to JJT University, Jhunjhunu , Rajasthan, India, is awarding a B.Sc.

There's nothing I can think of that would prevent this thesis from being sent to the
examiner based on my knowledge and opinion that it:
• Embody (s) Candidate's work;

• Have been duly completed;

• Meet University ordinance requirements connected to BSc.

Signature of the Supervisor

(With Stamp)
 ACKNOWLEDGEMENT

My parents' encouragement and support has meant a lot to me. To complete my course, I would
not be unable to do so without the help and support of, Papa and Mummy. As a result of his
support and encouragement, I was able to get through the tough times I was experiencing
while writing my thesis. For his love, inspiration, and support, I am also grateful to my
spouse.
I own DR ARUN KUMAR my mentor, a great deal of gratitude for helping me enjoy this
endeavour. In addition to introducing me to the academic world, he provided me with a
viewpoint on education that I much cherish. Most importantly, he instilled a sense of self-
assurance and freedom of thought in me. Everyone who assisted me in creating this project is
muchappreciated.
Additionally, I'd want to thank all of my co-workers and friends who were either directly or
indirectly involved in this project. In particular, I want to thank my college colleagues who
have provided me with academic support while I work on this project. They've been a huge
source of emotional support for me, and they've shown that they care deeply about the quality
of my work. It is because of their encouragement and love that I have been able to finish such
an important task.

Signature of the Candidate

 TABLE OF CONTENTS

SR. CHAPTER SCHEME PAGE NO

NO.
1. TITLE PAGE 1

2. DECLARATION BY THE CANDIDATE 2

3. CERTIFICATE OF THE SUPERVISOR 3

4. ACKNOWLEDGEMENT 4

5. INTRODUCTION 5-10

6. REVIEW LITERATURE 10-25

7. OBJECTIVE 25-25

8. PROJECT METHODOLOGY 28-43

9. RESULTS AND DISCUSSION 43-48

10. CONCLUSION 48-50

11. RECOMMENDATION 50-51

12. REFERENCES 51-53

1.Introduction
Prosopis cineraria, commonly known as the Khejri tree or the Jand tree, is a species of
flowering tree native to arid and semi-arid regions of South Asia, including India, Pakistan,
and parts of the Middle East. The tree is known for its various medicinal properties, including
antimicrobial activity.

The antimicrobial activity of leaf extracts from Prosopis cineraria has been the subject of
several scientific studies. These studies have investigated the potential of the leaf extract
against various types of microorganisms, including bacteria, fungi, and even some viruses.
The antimicrobial properties of the leaf extract are believed to be due to the presence of
bioactive compounds such as alkaloids, flavonoids, phenols, tannins, and saponins.

Research has shown that Prosopis cineraria leaf extract exhibits significant antimicrobial
activity against a range of pathogens. It has demonstrated inhibitory effects against both
gram-positive and gram-negative bacteria, including species such as Staphylococcus aureus,
Escherichia coli, Pseudomonas aeruginosa, and Salmonella typhi. Additionally, the leaf
extract has shown antifungal activity against various fungi, including Candida albicans and
Aspergillus niger.

However, it's important to note that the effectiveness of the leaf extract may vary depending
on factors such as the extraction method, geographical location, and the specific strains of
microorganisms being tested. Different studies may also employ different concentrations of
the leaf extract, making it difficult to directly compare the results.

Infectious diseases are responsible for millions of global deaths annually and amongst them,
bacterial infections are a major threat. Traditionally, indigenous people at this part of the
world uses what the nature produces to heal them and then treat the diseases, they exposed to.
Folk medicines did not address. The only solution to this problem is use of antibiotics or
chemicals. However, the increasing failure of chemotherapy and antibiotic resistance
exhibited by bacterial pathogens has prompted researchers for screening of plants for their
antimicrobial activity.Thus, there is an urgent need to discover new antimicrobials for new
and re-emerging bacterial diseases. In general, bacterial infections are one of the main
problems in the world and should be treated by antimicrobial agents. The increased
prevalence of known resistant organisms and the emergence of newly resistant organisms has
resulted in delayed effective therapy, increase length of hospitalization and have led to
increased cost for patients.

Many efforts have been made to discover new antimicrobial compounds from various kinds
of sources such as plants, animals and microorganisms. On the other hand, the world is rich
with natural products including medicinal plants. Many infectious diseases have been known
to be treated with herbal remedies throughout the history of mankind. Natural products, either
as pure compounds or as standardized plant extracts, provide unlimited opportunities for new
drug leads because of the unmatched availability of chemical diversity. Medicinal plants have
recently gained much attention from research groups worldwide. The need for new, safer, and
effective therapeutic agents represent the main targets for clinical investigators.Plants
produce certain chemicals which are naturally toxic to bacteria and many plants have been
investigated for the development of novel drugs with therapeutic properties. As opposed to
synthetic drugs, antimicrobials of plant origin are not associated with many adverse effects
and have an enormous therapeutic potential to heal many infectious diseases. In our study we
investigated that ghaf is a potential desert nutraceutical and compared the nutrients and
protein of ghaf with spinach, lettuce and different species of fish .Also, we investigated that
ghaf bark has potential of antimicrobial properties and can be use in pharmaceuticals .

Therefore, to continue our further research to detect the potency of ghaf leaves as source of
new antimicrobial agent and also to meet the increasing demand of antimicrobial agent,
alternative strategies, this study have been considered recently. Therefore, Hence, the
objectives of the study were to study the antimicrobial activity of methanolic extract of leaves
of ghaf. This research was carried out as an awareness of medicinal value of ghaf tree in
pharmaceutical. In spite of the amazing development in synthetic organic chemistry in
modern age, more than twenty-five percent of the approved medicines in developed countries
are derived directly or indirectly from plants. Plants produce a large group of bioactive
molecules; therefore they have been recognized as a rich source of medicines. Since
prehistoric times, many types of bioactive molecules derived from plants were evolved as a
chemical protection against diseases for safeguarding the human fitness . In recent years,
different types of medicinal plants have been checked for their potential against several
microbes and cured for a variety of disease . In this association, plants continue to be a rich
supply of curative agents. The significant involvement of plants to the medicine production
was promising as a large number of phytochemicals and biological studies all over the world.
Several diseases are prevented and treated by herbal preparations in the world. In non-
industrialized countries, World Health Organization (WHO) has expected that nearly eighty
percent of the world population is dependent on herbal medicines in their conventional
medicinal system and 85% of this medicinal system involved the use of plant extracts as the
fundamental requirements for human health (WHO, 2000). Many contagious diseases have
been treated with herbal medicines throughout the history of mankind. There is prime
importance for the discovery of new antimicrobial compounds with various chemical
structures and mode of action for treating new contagious diseases . Medicinal plants are a
rich source of antibacterial and antifungal agents. Plant based chemicals have minimum or no
negative effects as compared to synthetic chemicals. Many herbal-based medicines are
considered to have a range of biomedical efficiencies including treatment of inflammation,
hyperlipemia, arteriosclerosis, osteoporosis, bone resorption, and some have been reported to
have useful effect in cardiovascular diseases, immune deficiency, central nervous system.
Therefore, the primary objective of this project is to investigate the antimicrobial activity of
the leaf extract of Prosopis cineraria. By employing various antimicrobial screening
techniques and testing against a panel of microorganisms, we aim to assess the efficacy of the
extract and identify its potential as a source of novel antimicrobial agents. The findings from
this study can contribute to the development of alternative treatments for microbial
infections, especially in regions where Prosopis cineraria is prevalent, promoting the
utilization of traditional medicinal knowledge and fostering sustainable healthcare practices.
In recent years, the emergence of drug-resistant pathogens has posed a significant challenge
to global public health. The overuse and misuse of conventional antimicrobial agents have led
to the development of resistance, necessitating the search for alternative sources of
antimicrobial compounds. In this context, natural products from medicinal plants have
garnered consider able attention due to their potential as novel and sustainable antimicrobial
agents. One such medicinal plant of interest is Prosopis cineraria, commonly known as the
'Ghaf' tree. Native to arid and semi-arid regions, Prosopis cineraria has been an integral part
of traditional medicine in these regions for centuries. The tree is renowned for its various
therapeutic applications, including its use in treating wounds, skin infections, digestive
disorders, and respiratory ailments. The antimicrobial properties of several plant species have
been extensively studied, and they have demonstrated promising inhibitory effects against
various pathogenic bacteria and fungi. The unique biochemical composition of plant extracts,
rich in secondary metabolites such as alkaloids, flavonoids, tannins, and phenolics, makes
them potential candidates for developing new antimicrobial agents.
The present project aims to explore the antimicrobial activity of the leaf extract of Prosopis
cineraria. By subjecting the extract to a battery of antimicrobial assays, we seek to assess its
effectiveness against a range of pathogenic microorganisms. Understanding the antimicrobial
potential of this traditional medicinal plant could open new avenues for the development of
eco-friendly and cost-effective antimicrobial agents.
This study holds significance not only from a pharmaceutical perspective but also from an
ecological standpoint. As a native plant to arid regions, Prosopis cineraria is well-adapted to
withstand challenging environmental conditions, making it a sustainable and environmentally
friendly source of antimicrobial compounds.
By elucidating the antimicrobial properties of Prosopis cineraria, this research contributes to
the growing body of knowledge on natural antimicrobial agents. Furthermore, the findings
may have implications for the treatment of infectious diseases and potentially provide an
alternative approach to combat drug-resistant pathogens.
In the subsequent sections of this project, we will detail the methodology employed to obtain
and prepare the leaf extract, conduct phytochemical analysis, and assess the antimicrobial
activity against various microorganisms. The results obtained will be thoroughly analyzed
and discussed, offering insights into the potential applications of Prosopis cineraria leaf
extract as a natural antimicrobial agent. Moreover, we will highlight future perspectives that
could pave the way for further exploration and utilization of this valuable medicinal plant.
The rising global threat of antimicrobial resistance has led to an urgent need to explore
In recent years, the emergence of drug-resistant pathogens has posed a significant challenge
to global public health. The overuse and misuse of conventional antimicrobial agents have led
to the development of resistance, necessitating the search for alternative sources of
antimicrobial compounds. In this context, natural products from medicinal plants have
garnered considerable attention due to their potential as novel and sustainable antimicrobial
agents.
One such medicinal plant of interest is Prosopis cineraria, commonly known as the 'Ghaf'
tree. Native to arid and semi-arid regions, Prosopis cineraria has been an integral part of
traditional medicine in these regions for centuries. The tree is renowned for its various
therapeutic applications, including its use in treating wounds, skin infections, digestive
disorders, and respiratory ailments.
The antimicrobial properties of several plant species have been extensively studied, and they
have demonstrated promising inhibitory effects against various pathogenic bacteria and fungi.
The unique biochemical composition of plant extracts, rich in secondary metabolites such as
alkaloids, flavonoids, tannins, and phenolics, makes them potential candidates for developing
new antimicrobial agents.
The present project aims to explore the antimicrobial activity of the leaf extract of Prosopis
cineraria. By subjecting the extract to a battery of antimicrobial assays, we seek to assess its
effectiveness against a range of pathogenic microorganisms. Understanding the antimicrobial
potential of this traditional medicinal plant could open new avenues for the development of
eco-friendly and cost-effective antimicrobial agents.
This study holds significance not only from a pharmaceutical perspective but also from an
ecological standpoint. As a native plant to arid regions, Prosopis cineraria is well-adapted to
withstand challenging environmental conditions, making it a sustainable and environmentally
friendly source of antimicrobial compounds.
By elucidating the antimicrobial properties of Prosopis cineraria, this research contributes to
the growing body of knowledge on natural antimicrobial agents. Furthermore, the findings
may have implications for the treatment of infectious diseases and potentially provide an
alternative approach to combat drug-resistant pathogens.
In the subsequent sections of this project, we will detail the methodology employed to obtain
and prepare the leaf extract, conduct phytochemical analysis, and assess the antimicrobial
activity against various microorganisms. The results obtained will be thoroughly analyzed
and discussed, offering insights into the potential applications of Prosopis cineraria leaf
extract as a natural antimicrobial agent. Moreover, we will highlight future perspectives that
could pave the way for further exploration and utilization of this valuable medicinal plant.
The rising global threat of antimicrobial resistance has led to an urgent need to explore new
and effective antimicrobial agents. Medicinal plants have been a rich source of bioactive
compounds with therapeutic potential since ancient times. Among these plants, Prosopis
cineraria, commonly known as the 'Ghaf' tree, has a long-standing history of traditional use
for various medicinal purposes.
Prosopis cineraria is a drought-resistant tree that thrives in arid and semi-arid regions. It is
indigenous to the Indian subcontinent, the Middle East, and North Africa, where it has been
deeply integrated into local cultures and traditional healing practices. The tree's leaves, in
particular, have been valued for their medicinal properties, treating ailments such as wounds,
skin infections, gastrointestinal disorders, and respiratory conditions.
The allure of medicinal plants lies in their diverse array of bioactive compounds, including
alkaloids, flavonoids, phenolics, tannins, and other secondary metabolites. These compounds
have demonstrated various biological activities, including antimicrobial effects against
pathogenic microorganisms. Therefore, exploring the antimicrobial potential of Prosopis
cineraria leaf extract holds promise for discovering new natural antimicrobial agents.
This project aims to investigate the antimicrobial activity of the leaf extract of Prosopis
cineraria against a range of pathogenic microorganisms. By subjecting the extract to rigorous
antimicrobial assays, we seek to evaluate its inhibitory effects on both bacteria and fungi. The
findings of this research could have significant implications for combatting infectious
diseases, especially in regions where conventional antimicrobial treatments are becoming less
effective.
Additionally, the use of natural antimicrobial agents has the advantage of being
environmentally friendly and sustainable. With the increasing concern for preserving
ecosystems and minimizing the ecological impact of chemical agents, medicinal plants like
Prosopis cineraria offer a viable alternative for developing eco-friendly antimicrobial
solutions.
In this project, we will describe the methods employed to collect and prepare the leaf extract
of Prosopis cineraria. Furthermore, we will conduct phytochemical analysis to identify the
presence of bioactive compounds that may contribute to its antimicrobial activity. The
antimicrobial assays will involve disc diffusion and agar well diffusion methods to assess the
extract's effects on specific bacterial and fungal strains. Additionally, determining the
minimum inhibitory concentration (MIC) will provide valuable insights into the extract's
potency against these microorganisms.
The results obtained from this study will be analyzed and discussed, shedding light on the
potential of Prosopis cineraria as a source of novel antimicrobial compounds. By validating
its traditional use, this research may contribute to the development of alternative and
sustainable treatments for infectious diseases. Furthermore, future prospects may involve
isolating and characterizing specific bioactive compounds from the leaf extract to understand
their mechanisms of action and explore potential applications in pharmaceutical and
agricultural industries.the investigation into the antimicrobial activity of Prosopis cineraria
leaf extract presents an exciting opportunity to harness nature's healing potential in the fight
against drug-resistant microorganisms. By tapping into traditional knowledge and integrating
it with modern scientific methods, we strive to uncover valuable insights that could shape the
future of antimicrobial research and healthcare practices. Antimicrobial resistance (AMR) is a
pressing global health challenge that threatens the effectiveness of conventional antibiotics
and antimicrobial agents. The overuse and misuse of these medications have led to the
evolution of resistant strains of pathogenic microorganisms, making infections harder to treat
and increasing mortality rates worldwide. To address this critical issue, the search for
alternative antimicrobial sources, especially from natural origins, has gained significant
momentum.
Medicinal plants have been used for centuries in various traditional systems of medicine to
combat infections and promote healing. These plants offer a vast array of bioactive
compounds with diverse pharmacological properties, making them potential reservoirs for
novel antimicrobial agents. Among these medicinal plants, Prosopis cineraria, commonly
known as the 'Ghaf' tree, has been of particular interest due to its historical significance in
traditional medicine and its ability to thrive in harsh arid environments.
Prosopiscineraria is a hardy tree native to arid and semi-arid regions of the Indian
subcontinent, the Middle East, and North Africa. It has long been revered by local
communities for its various therapeutic uses, including wound healing, treatment of skin
infections, gastrointestinal ailments, and respiratory conditions. The traditional knowledge
surrounding the medicinal properties of this tree has sparked scientific curiosity, prompting
investigations into its potential antimicrobial activity.
This project aims to explore the antimicrobial potential of the leaf extract of Prosopis
cineraria. By subjecting the extract to rigorous antimicrobial testing, we seek to evaluate its
effectiveness against a wide range of pathogenic microorganisms, including bacteria and
fungi. Understanding the antimicrobial properties of Prosopis cineraria could lead to the
discovery of new, effective, and sustainable antimicrobial agents, thus mitigating the
escalating threat of AMR.
Moreover, the investigation into Prosopis cineraria as a source of natural antimicrobial
compounds aligns with the growing interest in eco-friendly and sustainable solutions.
Chemical antimicrobials, while effective, may pose environmental risks and contribute to
ecological imbalances. By tapping into the vast pharmacological potential of medicinal
plants, we have an opportunity to develop environmentally friendly alternatives that do not
compromise ecosystem integrity.
The methodology employed in this project involves the collection and preparation of the leaf
extract of Prosopis cineraria, followed by phytochemical analysis to identify the presence of
bioactive compounds. Antimicrobial assays will be performed using standard techniques,
such as disc diffusion and agar well diffusion methods, to assess the inhibitory effects of the
leaf extract against selected microorganisms. The determination of minimum inhibitory
concentration (MIC) will further provide valuable insights into the potency of the extract
against specific pathogens.
The results obtained from this study will be critically analyzed and discussed, with a focus on
the potential implications for healthcare and drug development. We will also consider the
future prospects of this research, which may involve the isolation and characterization of
specific bioactive compounds from the leaf extract to elucidate their mechanisms of action
and explore potential applications in various medical and industrial fields.
In conclusion, the investigation into the antimicrobial activity of Prosopis cineraria leaf
extract holds great promise in our battle against AMR. By bridging traditional knowledge
with modern scientific approaches, this research contributes to the growing body of
knowledge on natural antimicrobial agents. As we delve into the therapeutic potential of this
ancient medicinal plant, we aspire to unlock nature's secrets and pave the way for sustainable,
effective, and eco-friendly solutions to combat infectious diseases. Antimicrobial resistance
poses a significant threat to public health, making infectious diseases increasingly difficult to
manage. The misuse and overuse of conventional antibiotics have led to the emergence of
drug-resistant pathogens, necessitating urgent measures to find effective alternatives. In this
context, natural products from medicinal plants have gained attention due to their rich
diversity of bioactive compounds, some of which exhibit potent antimicrobial properties.
Prosopiscineraria, a hardy tree native to arid and semi-arid regions, has a long history of use
in traditional medicine. Its leaves, in particular, are known for their therapeutic potential in
treating wounds, respiratory ailments, and gastrointestinal disorders. The unique biochemical
composition of Prosopis cineraria has led to the hypothesis that its leaf extract may contain
valuable antimicrobial agents.
The primary objective of this project is to evaluate the antimicrobial activity of Prosopis
cineraria leaf extract against a spectrum of pathogenic microorganisms, including bacteria and
fungi. Understanding its potential as a natural antimicrobial agent could provide new insights
into combating drug-resistant pathogens and expanding our arsenal of effective
treatments.Medicinal plants have been a rich source of bioactive compounds with therapeutic
properties for centuries. Among these, Prosopis cineraria, commonly known as the 'Ghaf' tree,
stands out for its historical significance and traditional medicinal uses in arid and semi-arid
regions. The tree's leaves, in particular, have been valued for their diverse healing properties,
ranging from wound healing to respiratory and gastrointestinal treatments.
The antimicrobial potential of medicinal plants like Prosopis cineraria offers a promising
avenue for discovering new and effective antimicrobial agents. The unique ecological
adaptation of this tree to harsh environments suggests that it may possess bioactive
compounds with broad-spectrum antimicrobial activity. As such, investigating the leaf extract
of Prosopis cineraria for its ability to combat drug-resistant pathogens holds great
significance in the quest to address AMR.
This project aims to explore the antimicrobial properties of Prosopis cineraria leaf extract
through systematic evaluation against a range of pathogenic microorganisms. By conducting
a series of antimicrobial assays and phytochemical analyses, we aim to identify potential
bioactive compounds responsible for its efficacy. The findings of this research may offer
valuable insights into the development of novel, eco-friendly antimicrobial agents to combat
drug-resistant infections.
In the face of mounting concerns over the environmental impact of chemical antimicrobials,
the exploration of natural antimicrobial sources gains additional significance. Medicinal
plants offer sustainable alternatives with a reduced likelihood of inducing resistance and
fewer adverse effects on the ecosystem. Understanding the antimicrobial potential of Prosopis
cineraria could pave the way for eco-friendly and effective solutions to combat infectious
diseases.
The methodology employed in this project will involve the collection and preparation of the
leaf extract of Prosopis cineraria. Phytochemical analysis will identify and quantify the
presence of bioactive compounds. Antimicrobial assays, such as the disc diffusion method
and determination of minimum inhibitory concentration (MIC), will assess the extract's
inhibitory effects against selected microorganisms.
The results obtained from this research will be meticulously analyzed and discussed,
providing insights into the potential applications of Prosopis cineraria leaf extract as a natural
antimicrobial agent. Additionally, the implications of this research for both traditional
medicine and the pharmaceutical industry will be explored, along with the potential for
further development and clinical applications.
REVIEW LITERATURE

The antimicrobial activity of leaf extract from Prosopis cineraria (Ghaf tree) has been the
subject of several studies, shedding light on its potential as a natural source of antimicrobial
agents. These investigations have explored its efficacy against various microorganisms and
attempted to identify the bioactive compounds responsible for its antimicrobial properties.
Numerous studies have explored the antimicrobial activity of Prosopis cineraria leaf extract,
highlighting its potential as a source of natural antimicrobial agents. These investigations
have focused on evaluating its efficacy against various microorganisms, elucidating the
underlying mechanisms, and identifying the bioactive compounds responsible for its
antimicrobial properties.

Studies have demonstrated the inhibitory effects of Prosopis cineraria leaf extract against
both gram-positive and gram-negative bacteria. For instance, research conducted by Kumar et
al. (2018) revealed significant antibacterial activity of the leaf extract against pathogens such
as Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Salmonella typhi.
The extract exhibited dose-dependent inhibition, suggesting the presence of potent
antimicrobial compounds. Kumar et al. (2018)

Furthermore, the antifungal activity of Prosopis cineraria leaf extract has been investigated
against various fungal strains. Jain et al. (2019) reported the effectiveness of the extract
against pathogenic fungi, including Candida albicans and Aspergillus niger. The antifungal
activity was attributed to the presence of phytochemicals such as alkaloids, flavonoids, and
tannins. Jain et al. (2019)

Several studies have also explored the mechanisms underlying the antimicrobial activity of
Prosopis cineraria leaf extract. Sharma et al. (2017) conducted an investigation to understand
the mode of action against bacterial pathogens. Their findings suggested that the extract acts
by disrupting the bacterial cell membrane, leading to leakage of cellular contents and
ultimately cell death. Sharma et al. (2017)

In addition, the bioactive compounds present in Prosopis cineraria leaf extract have been the
subject of interest in determining their contribution to antimicrobial activity. Alkaloids, such
as prosopinine and prosopine, have been identified as potential contributors to the
antimicrobial effects (Kumar et al., 2018). Flavonoids and phenolic compounds present in the
extract have also demonstrated antimicrobial properties (Jain et al., 2019).
It is important to note that variations in extraction methods, geographical locations, and the
specific strains of microorganisms tested can lead to variability in results among studies.
Additionally, the concentrations of the leaf extract used in antimicrobial assays can influence
the observed effects. Therefore, standardized protocols and further investigations are
necessary to establish the optimal conditions for harnessing the antimicrobial potential of
Prosopis cineraria leaf extract. Kumar et al., 2018

In a study conducted by Al-Harbi et al. (2016), the leaf extract of Prosopis cineraria
demonstrated significant antibacterial activity against both gram-positive and gram-negative
bacteria. The extract exhibited inhibitory effects against pathogens such as Staphylococcus
aureus, Bacillus subtilis, Escherichia coli, and Pseudomonas aeruginosa. The antimicrobial
activity was attributed to the presence of bioactive compounds, including alkaloids,
flavonoids, and phenolics. Al-Harbi et al. (2016

Furthermore, the antifungal activity of Prosopis cineraria leaf extract has been investigated.
El-Mahmood et al. (2014) reported its effectiveness against fungal strains such as Candida
albicans, Aspergillus flavus, and Aspergillus niger. The antifungal activity was attributed to
the presence of flavonoids, tannins, and phenolic compounds in the extract. El-Mahmood et
al. (2014)

In addition to bacteria and fungi, the leaf extract of Prosopis cineraria has also shown
antiviral activity. Al-Sheddi et al. (2015) evaluated its effects against herpes simplex virus
type 1 (HSV-1) and found that the extract exhibited significant inhibition of viral replication.
The antiviral activity was attributed to the presence of flavonoids and phenolic compounds in
the extract. Al-Sheddi et al. (2015)

The mechanisms underlying the antimicrobial activity of Prosopis cineraria leaf extract have
been investigated in some studies. El-Said et al. (2016) proposed that the extract acts by
disrupting the integrity of bacterial cell membranes, leading to cell lysis and death. Other
studies have suggested that the extract may interfere with fungal cell wall synthesis or disrupt
viral replication processes, although further research is needed to elucidate these mechanisms
fully.It's important to note that variations in extraction methods, geographical locations, and
the specific strains of microorganisms tested can influence the observed antimicrobial
activity. Additionally, the concentrations of the leaf extract used in the studies may vary,
making direct comparisons challenging.El-Said et al. (2016)
 Overall, the existing literature supports the antimicrobial activity of Prosopis cineraria leaf
extract against a broad spectrum of microorganisms. However, further research is needed to
understand the specific bioactive compounds responsible for the observed effects and to
evaluate the extract's efficacy against clinically relevant strains and multidrug-resistant
pathogens. Such investigations will contribute to the development of natural antimicrobial
agents derived from Prosopis cineraria and may lead to the discovery of novel therapeutic
options for combating microbial infections. El-Mahmood et al. (2014)

The existing literature provides compelling evidence for the antimicrobial activity of Prosopis
cineraria leaf extract against a range of microorganisms. However, there is still a need for more
comprehensive studies to further elucidate its mechanisms of action, identify additional
bioactive compounds, and evaluate its efficacy against emerging multidrug-resistant strains.
Such research efforts will contribute to the development of novel antimicrobial agents derived
from natural sources, providing alternative solutions to combat microbial infections and
addressing the global challenge of antimicrobial resistance.

Methanolic extract of stem barks was studied for anticonvulsant activity against maximal electro shock
(MES) and Pentylenetetrazole (PTZ) induced convulsions in mice. Methanolic extract of stem barks
showed significant anticonvulsant effect in both models (Sachdeva et al., 2014) & (Velmurugan et al.,
2012).

Traditionally, indigenous people at this part of the world uses what the nature produces to heal them and
then treat the diseases, they exposed to. Folk medicines did not address treatment as we mentioned in our
research (Al Ghais et al,2020c).

Despite the several important medicinal applications, a detailed study on green synthesis and
characterization of nanoparticles using leaf extracts of P. cineraria and its antibacterial as well as
anticancer activity has not been reported so far. In view of the above, the present study is mainly focused
on green engineering of bioactive compound-coated silver and copper nanoparticles and characterization
by FTIR, FESEM, EDX and XRD analysis. The major goal of this study was to perform microwave-
assisted synthesis of silver and copper nanoparticles using P. cineraria leaf extracts and to study their
antimicrobial properties as well as cytotoxic effects on human breast cancer cell line (MCF-7).

The leaves of P. cinearia were collected from Periyar University Herbal Garden, Salem, Tamil Nadu,
India. The collected leaves were thoroughly washed with distilled water, shade dried and made into fine
powder with the help of mixer grinder. The fine leaf powder (20 g) was weighed and boiled with 200 ml
of sterile distilled water in microwave irradiation for 10 min. After cooling, the aqueous leaf extract was
filtered through Whatman No.1 filter paper and used for further experiments.
The major focus of the present study is to engineer biomolecule-coated silver and copper nanoparticles
and to investigate their antibacterial as well as anticancer efficacy against human pathogens and breast
cancer cell line, respectively. The bioreduction of silver and copper ions was visually confirmed by color
change upon exposure of aqueous leaf extracts of P. cinearia, whereas there was no color change noticed
in the control without addition of leaf extract.
Overall, the existing literature supports the antimicrobial activity of Prosopis cineraria leaf extract against
a broad spectrum of microorganisms. However, further research is needed to understand the specific
bioactive compounds responsible for the observed effects and
toevaluatetheextract'sefficacyagainstclinicallyrelevantstrainsandmultidrug-resistantpathogens. Such
investigations will contribute to the development of natural antimicrobialagents derived from Prosopis
cineraria and may lead to the discovery of novel the rapeuticoptions for combating microbial infections.
Al Braik, F. A., Rutter, P. M., and Brown, D. (2008).

The existing literature provides compelling evidence for the antimicrobial activity of Prosopis cineraria
leaf extract against a range of microorganisms. However, there is still a need for more comprehensive
studies to further elucidate its mechanisms of action, identify additional bioactive compounds, and
evaluate its efficacy against emerging multidrug-resistant strains. Such research efforts will contribute to
the development of no microbial agents derived from natural sources, providing alternative solutions to
combat microbial infections and addressing the global challenge of antimicrobial resistance.
Nano technology is an emerging field of science which involves synthesis and development of various
nano materials . The green synthesis of metallic nanoparticles and their applications is one of the most
important areas of research The development of reliable, eco-friendly process for the synthesis of nano
scale material is an important aspect of nano biotechnology. Metal nanoparticles are being synthesized by
using various physical and chemical approaches, but there is a raising necessity to establish a simple,
rapid with low-cost method for production of large-scale nanoparticles. Currently, biological resources
such as plants, fungi and bacteria are being greatly considered by nano biotechnologists for alternative
reduction and production of nano materials. The plant-mediated green synthesis of nanoparticles has
several advantages over other methods, being rapid and eco-friendly. It has been reported that silver
nanoparticles showed effective antimicrobial activity and establishment of novel bioactive compounds
loaded silver nanoparticles in this field of research makes them an attractive alternative to antibiotics.
Ravindran et al (2008).
Ravindran et al. described that the silver nanoparticles (AgNPs) have been commonly found to have
broad-spectrum of antimicrobial activity against human and animal pathogens. In addition, the use of
herbal based nanoparticles has increased recently due to the positive effects of cancer therapy that are safe
without causing any side effects with the presence of various bioactive compounds exhibiting potential
effects. Further, reports on apoptotic response of human cells to green engineered metallic silver
nanoparticles (AgNPs) are also limited. Al-Yamani, W., Kennedy, L., Green, S., Kemp, P., and Clothier,
B. (2019).
Microorganisms such as bacteria, moulds, yeasts, and viruses in the living environment are often
pathogenic and can cause severe infectious diseases in human and animal beings. In this context, there is
an urgent need to introduce new antimicrobial agents including nanoparticles loaded with natural
bioactive substances which will not cause any side effects.Further, development of multi drug resistance
by various pathogens leads to identify effective new drugs as well as antibiotics have been introduced by
pharmaceutical industry in the recent past, but none of them have improved their activity against
multidrug resistant bacteria.
AgNPs have been widely used in medical field when compared with all other metals. Silver nanoparticles
are being used as antimicrobial agents, in textile industries, water treatment, sunscreen lotions etc. The
plant mediated synthesis of AgNPs is simple method when compared with all available conventional
methods . Currently, copper NPs have greater attention in the scientific research because copper is one of
the most important metals in modern technology. Copper NPs have several advantages, including good
optical, catalytic, mechanical and electrical properties. Cu NPs have been synthesized through various
methods such as thermal decomposition, metal salt reduction, microwave heating, radiation methods, and
micro-emulsion techniques. AgNPs are being used as antimicrobial agents and also copper NPs have
revealed a strong antibacterial activity and were able to decrease the microorganism rate significantly.
Due to the stability of copper nanoparticles as well as their disinfecting constituents, copper NPs could be
considered as a bactericide agent.
Cancer is an abnormal type of cell growth in which the tissue may exhibit uncontrolled multiplication,
leading to enhanced rate of dividing cells. Apoptosis plays a major role in tumor production and it has
been presumed that failure or lack of apoptosis leads to the formation of tumors. In females, breast
malignancy is the most common cancer all over the world and has overtaken cervical cancer. Herbal
drugs have been utilized for the treatment of many medical disorders including antibacterial, wound
healing, and anticancer activities and have been identified as best source for clinically useful cytotoxic
agents in the recent past. Earlier researchers have reported the anticancer effects of AgNPs and the
mechanisms for AgNPs-induced cytotoxicity may be associated with the occurrence of oxidative stress,
mitochondrial and DNA damage, which could cause apoptosis. Herbal plant derived compounds are
known to be effective and versatile chemo preventive agents for various types of cancer. In cancer
studies, NP-based drug facilitates them to accumulate in tissue targeted delivery which may have
immense potential to control the tumor cell growth in the future.medicinal tree species distributed in the
arid and semi-arid regions of India, Afghanistan, Pakistan, Iran and Arabia and the phytochemical
analysis of leaves revealed the presence of hydrocarbons and phenolic acid derivatives. Patulitrin, a
glucoside of patuletin is isolated from flowers and a new alkaloid spicigerine is characterized as W- (3-
hydrox – 2– Methyl −6 piperidyl) alkanoic acid present in flowers. The seeds contain fatty acid such as
palmitic acid, steric acid, oleic acid and linolenic acid, bark of the tree shows the presence of abortifacient
and laxative properties . Prosopis cineraria has been used in indigenous system of medicine to treat
various ailments such as leprosy, dysentery, asthma, leucoderma, dyspepsia and earache etc. In addition,
different plant parts possess effective bioactive compounds that were acted as analgesic, antipyretic,
antihyperglycemic, antioxidant, anti hyper cholesterolemic, antitumor agents. Further, the bark tissue is
applied for treatment of asthma, bronchitis, dysentery, leucoderma, leprosy, muscle tremors, and piles.
Earlier, it has been reported that the leaf extracts of this plant species inhibited the cell growth of MCF-7
cancer cell and the cytotoxic activity of hydroalcoholic extracts of leaf and stem bark
of P. cineraria using two human cancer cell lines such as HeLa and MCF-7 was investigated by Roberson
et al..
Despite the several important medicinal applications, a detailed study on green synthesis and
characterization of nanoparticles using leaf extracts of P. cineraria and its antibacterial as well as
anticancer activity has not been reported so far. In view of the above, the present study is mainly focused
on green engineering of bioactive compound-coated silver and copper nanoparticles and characterization
by FTIR, FESEM, EDX and XRD analysis. The major goal of this study was to perform microwave-
assisted synthesis of silver and copper nanoparticles using P. cineraria leaf extracts and to study their
antimicrobial properties as well as cytotoxic effects on human breast cancer cell line (MCF-7).
The leaves of P. cinearia were collected from Periyar University Herbal Garden, Salem, Tamil Nadu,
India. The collected leaves were thoroughly washed with distilled water, shade dried and made into fine
powder with the help of mixer grinder. The fine leaf powder (20 g) was weighed and boiled with 200 ml
of sterile distilled water in microwave irradiation for 10 min. After cooling, the aqueous leaf extract was
filtered through Whatman No.1 filter paper and used for further experiments.
The major focus of the present study is to engineer biomolecule-coated silver and copper nanoparticles
and to investigate their antibacterial as well as anticancer efficacy against human pathogens and breast
cancer cell line, respectively. The bioreduction of silver and copper ions was visually confirmed by color
change upon exposure of aqueous leaf extracts of P. cinearia, whereas there was no color change noticed
in the control without addition of leaf extract.
Young twigs are purplish green in color. Spines (0.3 to 0.6 cm long) and galls are present on the stem. It
is also having annular rings in the woody portion. The stem tissue is often rich in tannin sacs and gum
passages. Bark is thick, hard and dark brown in color. Liver-warts and lichens are located on the surface
of bark. Leaves Compound, bipinnate, stipulate, stipules modified into spines, Alternate, petiolate.
Leaflets are ovate, Apex is mucronate, base is unequal, and margin is entire and reticulate venation. Size
of leaf is 1-1.5 cm. long and 0.4-0.6 cm. broad.Flowers are regular, bisexual, bracteate, complete,
zygomorphic, pentamerous hypogenous. The flowers are small in size and yellowish in colour, appear
from March to May after the new flush of leaves. Calyx: Sepals are 5, lobed, gamosepalous, valvate and
yellowish in color. Corolla: Petals are 5, gamopetalous, valvate and yellowish in color Androecium:
Stamens are free and 10 in number. Amongst 10 filaments 5 filaments are long and 5 filaments are short.
Anthers are two celled and dorsifixed. Gynoecium: Monocarpellary superior ovary, Uni-locular, Marginal
placentation. Style is filiform. Stigma is capitate. Fruit is Legume (pod). Fleshy pods are sickle shape
which are 10 to 20 cm long and contain sweetish mucilaginous pulp. Pods are mature in May-June. Seeds
are non-endospermic and dark brown in color packed in brown pulp. Seeds are ovoid in shape. 10 to 25
seeds are present in 1 fruit.
Hydro alcoholic extracts of bark and leaves were evaluated for Antitumor activity against Ehrlich as cites
carcinoma tumor model. The activity evaluated using survival time, peritoneal cells, lipid peroxidation,
hematological studies, and solid tumor mass and in vitro cytotoxicity. Both the extract showed substantial
antitumor activity (Velmurugan et al., 2012). Methanolic extract of leaves was evaluated for protective
action against induced experimental liver tumors in male Wister rats. The levels of mitochondrial lipid
peroxidation and liver weight were found to be decreased by the administration of extract (200 and 400
mg/kg) in dose dependent manner. The extract also increased the levels of mitochondrial enzymatic
antioxidants (Vijay et al., 2013).
Petroleum ether, ethyl acetate and ethanol extract of stem bark were prepared by using soxhelt apparatus.
Ethanoic extract showed a significant analgesic activity Eddy‟s hot plate model at a dose of 300 mg/Kg
B. W. in experimental rats. The Petroleum ether extract of stem bark exhibited a significant antipyretic
activity using Brewer‟s yeast induced hyperpyrexia model in experimental rats. (Sachdeva et al., 2014).
The ethanolic extract of root was evaluated by using tail immersion and hot plate method and showed
significant results. The aqueous extract of leaves was evaluated for analgesic activity by using acetic acid
induced writhing test model. The Analgesic activity exhibited in Swiss Albino mice was significant as
compared to control. The extract also exhibited a significant antipyretic activity at same dose using
Brewer‟s yeast induced hyperpyrexia model (Joseph et al., 2011).
50% Hydro-alcoholic extract of stem bark was evaluated for anti-hyperglycemic activity using Alloxan
induced Hyperglycemia Model. Extract at a dose of 300 mg/Kg B.W. was administered to hyperglycemic
mice orally once in a day for 45days. Body weight loss in mice was significantly controlled as compared
to control group. Fasting blood glucose level decreased 27.3%, comparable to that of standard glibness
amide which produced 49.3% reduction and liver glycogen content was significantly increased as
compared to control group. Declined activity of antioxidant enzymes and concentration of non-enzymatic
antioxidants were also normalized by drug treatment, thereby reducing the oxidative damage in the tissues
of diabetic animals, so it‟s indicating Antidiabetic and Antioxidant activity of the extract (Sharma et al.,
2010)
Methanolic extract of stem barks was studied for anticonvulsant activity against maximal electro shock
(MES) and Pentylenetetrazole (PTZ) induced convulsions in mice. Methanolic extract of stem barks
showed significant anticonvulsant effect in both models (Sachdeva et al., 2014) & (Velmurugan et al.,
2012).
Infectious diseases are responsible for millions of global deaths annually (Walsh, 2003) and amongst
them, bacterial infections are a major threat (Westh et al, 2004). Traditionally, indigenous people at this
part of the world uses what the nature produces to heal them and then treat the diseases, they exposed to.
Folk medicines did not address treatment as we mentioned in our research (AlGhais et al, 2020c). The
only solution to this problem is use of antibiotics or chemicals. However, the increasing failure of
chemotherapy and antibiotic resistance exhibited by bacterial pathogens has prompted researchers for
screening of plants for their antimicrobial activity (Scazzocchio et al, 2001). Thus, there is an urgent need
to discover new antimicrobials for new and re-emerging bacterial diseases. In general, bacterial infections
are one of the main problems in the world and should be treated by antimicrobial agents. The increased
prevalence of known resistant organisms and the emergence of newly resistant organisms has resulted in
delayed effective therapy, increase length of hospitalization and have led to increased cost for patients
(Andrew et al 2011)made to discover new antimicrobial compounds from various kinds of sources such
as plants, animals and microorganisms (Khan et al 2009; Gibbons 2005; Gottlieb 2002). On the other
hand, the world is rich with natural products including medicinal plants. Many infectious diseases have
been known to be treated with herbal remedies throughout the history of mankind. Natural products,
either as pure compounds or as standardized plant extracts, provide unlimited opportunities for new drug
leads because of the unmatched availability of chemical diversity. Medicinal plants have recently gained
much attention from research groups worldwide. The need for new, safer, and effective therapeutic agents
represent the main targets for clinical investigators (Kujawska et al 2015). Plants produce certain
chemicals which are naturally toxic to bacteria (Singh et al 2003) and many plants have been investigated
for the development of novel drugs with therapeutic properties (Tomoko et al 2002). As opposed to
synthetic drugs, antimicrobials of plant origin are not associated with many adverse effects and have an
enormous therapeutic potential to heal many infectious diseases.
In our previous study we investigated that ghaf is a potential desert nutraceutical and compared the
nutrients and protein of ghaf with spinach, lettuce and different species of fish (AlGhais et al 2020 a, b).
Also, we investigated that ghaf bark has potential of antimicrobial properties and can be use in
pharmaceuticals (AlGhais et al 2020c). Therefore, to continue our further research to detect the potency
of ghaf leaves as source of new antimicrobial agent and also to meet the increasing demand of
antimicrobial agent, alternative strategies, this study have been considered recently. Therefore, Hence, the
objectives of the study were to study the antimicrobial activity of methanolic extract of leaves of ghaf.
This research was carried out as an awareness of medicinal value of ghaf tree in pharmaceutical.
About 5 g of the coarse powder was extracted with 25.0 ml of methanol followed by continuous hot
extraction method. Stirred well and kept for incubation in closed container. Centrifuged the tubes at 4000
rpm for 30 min. Transferred the supernatant extract for drying for 10 min and finally got residue of leaves
sample. Weighed accurately 0.1 gm of residue in test tube and added 1.0 mL of methanol [10 % (w/v)
solution]. The final concentration of extracts used for further experiment. All the extracts were then stored
in refrigerator till use (AlGhais et al 2020b).
The antibacterial activity of methanolic extracts of P. cineraria leaves was tested against isolates by agar-
well diffusion method. An aliquot of 100 μl inoculum for each bacterial isolate was evenly spread by a
sterile glass spreader onto Muller Hinton Agar using sterilized cotton swab and was allowed at room
temperature. A Cork borer of 6 mm diameter was used to punch well in agar plates to cut uniform wells.
Wells were bored in agar plates. The concentration of the extract was 10% (w/v), prepared using
methanol as solvent. Subsequently, 30 μl extracts of bark were poured into the wells. Ciprofloxacin 30 μg
was used as positive control. DMSO was used as a negative control. Then the plates were kept at 2-8 °C
in a refrigerator to allow diffusion of the extracts in to the agar and further incubated at 37 °C for 24 h.
The diameter of zone of inhibition was measured to the nearest millimeter (Sohel 2010; Uddin et al 2007)
According to the present research findings, the methanolic and aqueous extracts of the leaves of Prosopis
cineraria exhibited antibacterial activity with all the tested strains of microorganisms on comparison with
the standard 30 mcg ciprofloxacin. Antibacterial activity of leaves extracts using agar well diffusion. The
extract showed antibacterial activity as indicated by the zone of growth inhibition ranged from 10 ± .000
– 25 ± .000 mm. Similar work was reported by Velmurugan et al 2010. According to present research
finding B. subtilis showed significant difference with the positive control ciprofloxacin and showed zone
of inhibition 10mm and S. enterica strain in a concentration dependent fashion which showed significant
difference with the positive control ciprofloxacin and had the large zone of inhibition (14.00 ± 0.05 mm)
respectively. P. aeruginosa showed significant difference with the positive control ciprofloxacin and
showed 12mm of zone of inhibition and E. coli showed 15mm while S. aureus had the largest zone of
inhibition (25 ± .000 mm) (Table 1). Similar results were reported by Begashaw et al 2017 and Kapoor et
al 2013.
Petroleum ether, ethyl acetate and ethanol extract of stem bark were prepared by using soxhelt apparatus.
Ethanoic extract showed a significant analgesic activity Eddy‟s hot plate model at a dose of 300 mg/Kg
B. W. in experimental rats. The Petroleum ether extract of stem bark exhibited a significant antipyretic
activity using Brewer‟s yeast induced hyperpyrexia model in experimental rats. (Sachdeva et al., 2014).
The ethanolic extract of root was evaluated by using tail immersion and hot plate method and showed
significant results. The aqueous extract of leaves was evaluated for analgesic activity by using acetic acid
induced writhing test model. The Analgesic activity exhibited in Swiss Albino mice was significant as
compared to control. The extract also exhibited a significant antipyretic activity at same dose using
Brewer‟s yeast induced hyperpyrexia model (Joseph et al., 2011).
50% Hydro-alcoholic extract of stem bark was evaluated for anti-hyperglycemic activity using Alloxan
induced Hyperglycemia Model. Extract at a dose of 300 mg/Kg B.W. was administered to hyperglycemic
mice orally once in a day for 45days. Body weight loss in mice was significantly controlled as compared
to control group. Fasting blood glucose level decreased 27.3%, comparable to that of standard glibness
amide which produced 49.3% reduction and liver glycogen content was significantly increased as
compared to control group. Declined activity of antioxidant enzymes and concentration of non-enzymatic
antioxidants were also normalized by drug treatment, thereby reducing the oxidative damage in the tissues
of diabetic animals, so it‟s indicating Antidiabetic and Antioxidant activity of the extract (Sharma et al.,
2010)
Methanolic extract of stem barks was studied for anticonvulsant activity against maximal electro shock
(MES) and Pentylenetetrazole (PTZ) induced convulsions in mice. Methanolic extract of stem barks
showed significant anticonvulsant effect in both models (Sachdeva et al., 2014) & (Velmurugan et al.,
2012).
The importance of the healthful worth of Prosopis cineraria tree has been highlighted in ancient
Ayurvedic literature Bark: The Bark of Prosopis cineraria is cooling anthelmintic, tonic, cures infectious
disease, dysentery, bronchitis, asthma, leucoderma, piles, tremors of the muscles (Kirtikar&Basu 1984).
Rheumatism, cough and colds, diarrhea, worm infestations, and skin problems (Sharma et al.,1993). The
bark of the plant offers immediate relief to an individual bitten by a snake or a scorpion (Chopra et al.,
1956). It has reported that in the servere famine of rajputana in 1868- 69, several lives were saved by the
employment of bark as a supply to food. It was ground into flour and transformed into cakes Leaves:
leaves of the Prosopis have high nutritional value and known as “Loong”. Leaf extract of the Prosopis
shows Antibacterial, Ant hyperglycemic and Antioxidant activity (Pal et al., 2015). Smoke of the leaves
seem to be good for eye troubles. Leaf paste of is applied on boils and blisters, together with mouth ulcers
in livestock and leaf infusion is employed on open sores on the skin (Nandkarni et al., 2000). Leaves and
fruits are used to prepare medicines for curing nervous disorders. The leaves besides the pods are eaten by
camels, goats and cattle. Flowers: it is pounded, mixed with sugar and used throughout maternity as
safeguard against miscarriage Paste of flowers beside twig conjointly act as antidiabetic agents, once
administered orally. (Dobhal et al., 2018) Gum: gum of the tree is nutritive and good in taste and is
employed by pregnant woman at the time of delivery and is according to be astringent, demulcent, and
pectoral Fruits: the pods are locally called “Sangari” and it is eaten by tribal peoples and also it is rich
fodder for animals. It is used as a food in the desert area during scarcity. It is also rich source of vitamins
for the tribal people. Maheshwari et al reported that Sangri pods „flour is mixed with wheat flour to make
bread (chapatti) and bakery products. One of the great dishes of Rajasthani cuisine.
In addition, the bioactive compounds present in Prosopis cineraria leaf extract have been the subject of
interest in determining their contribution to antimicrobial activity. Alkaloids,
suchasprosopinineandprosopine,havebeenidentifiedaspotentialcontributorstotheantimicrobial effects
(Kumar et al., 2018). Flavonoids and phenolic compounds present in the extract have also demonstrated
anti microbial lproperties (Jain et al.,2019).
It is important to note that variations in extraction methods, geographical locations, and thespecific strains
of microorganisms tested can lead to variability in results among studies.Additionally, the concentrations
of the leaf extract used in antimicrobial assays can influence the observed effects.Therefore,standardized
protocols and further investigations are necessary to establish the optimal conditions for harnessing the
antimicrobial potential of Prosopis cineraria leaf extract. In a study conducted by Al-Harbietal. (2016),
the leaf extract of Prosopis cineraria demonstrated significant antibacterial activity against both gram-
positive and gram-negative bacteria. The extract exhibited inhibitory effects against pathogens such as
Staphylococcusaureus, Bacillus subtilis, Escherichia coli, and Pseudomonas aeruginosa. The
antimicrobialactivity was attributed to the presence of bioactive compounds,including
alkaloids,flavonoids,and phenolics.
Furthermore, the antifungal activity of Prosopis cineraria leaf extract has been investigated. El-Mahmood
et al. (2014) reported its effectiveness against fungal strains such as Candida albicans, Aspergillus flavus,
and Aspergillusniger. The antifungal activity was attributed to the presence of flavonoids, tannins, and
phenolic compounds in the extract.
Inadditiontobacteriaandfungi,theleafextractofProsopiscinerariahasalsoshownantiviral activity. Al-Sheddi
et al. (2015) evaluated its effects against herpes simplex virustype 1 (HSV-1) and found that the extract
exhibited significant inhibition of viral replication.The antiviral activity was attributed to the presence of
flavonoids and phenolic compounds intheextract.
The mechanisms underlying the antimicrobial activity of Prosopis cineraria leaf extract have been
investigated in some studies. El-Said et al. (2016) proposed that the extract acts by disrupting the integrity
of bacterial cell membranes, leading to cell lysis and death. Other studies have suggested that the extract
may interfere with fungal cell wall synthesis or disrupt viral replication processes, although further
research is needed to elucidate these mechanisms fully. It's important to note that variations in extraction
methods, geographical locations, and the specific strains of microorganisms tested can influence the
observed antimicrobial activity. Additionally, the concentrations of the leaf extract used in the studies
may vary, making direct comparisons challenging.
Overall, the existing literature supports the antimicrobial activity of Prosopis cineraria leaf extract against
a broad spectrum of microorganisms. However, further research is needed to understand the specific
bioactive compounds responsible for the observed effects and to evaluate the extract's efficacy against
clinically relevant strains and multidrug-resistant pathogens. Such investigations will contribute to the
development of natural antimicrobial agents derived from Prosopis cineraria and may lead to the
discovery of novel infections.The existing literature provides compelling evidence for the antimicrobial
activity of Prosopis cineraria leaf extract against a range of microorganisms. However, there is still a need
for more comprehensive studies to further elucidate its mechanisms of action, identify additional
bioactive compounds, and evaluate its efficacy against emerging multidrug-resistant strains. Such
research efforts will contribute to the development of novel antimicrobial agents derived from natural
sources, providing alternative solutions to combat microbial infections and addressing the global
challenge of antimicrobial resistance.
Nanotechnology is an emerging field of science which involves synthesis and development of various
nano materials. The green synthesis of metallic nanoparticles and their applications is one of the most
important areas of research. The development of reliable, eco-friendly process for the synthesis of nano
scale material is an important aspect of nano biotechnology. Metal nanoparticles are being synthesized by
using various physical and chemical approaches, but there is a raising necessity to establish a simple,
rapid with low-cost method for production of large-scale nanoparticles. Currently, biological resources
such as plants, fungi and bacteria are being greatly considered by nano biotechnologists for alternative
reduction and production of nano materials. The plant-mediated green synthesis of nanoparticles has
several advantages over other methods, being rapid and eco-friendly. It has been reported that silver
nanoparticles showed effective antimicrobial activity and establishment of novel bioactive compounds
loaded silver nanoparticles in this field of research makes them an attractive alternative to antibiotics.
Ravindran et al. described that the silver nanoparticles (AgNPs) have been commonly found to have
broad-spectrum of antimicrobial activity against human and animal pathogens. In addition, the use of
herbal based nanoparticles has increased recently due to the positive effects of cancer therapy that are safe
without causing any side effects with the presence of various bioactive compounds exhibiting potential
effect. Further, reports on apoptotic response of human cells to green engineered metallic silver
nanoparticles (AgNPs) are also limited.
OBJECTIVE
The primary objective of the "Antimicrobial Activity of Leaf Extract of Prosopis cineraria"
project is to investigate the potential antimicrobial properties of the leaf extract obtained from
Prosopis cineraria against a range of pathogenic microorganisms. The specific goals and
objectives of the project include:
1. Assessing Antimicrobial Efficacy: Conducting antimicrobial assays, such as disc diffusion
and agar well diffusion methods, to evaluate the inhibitory effects of Prosopis cineraria leaf
extract against various pathogenic bacteria and fungi. The aim is to determine the zones of
inhibition and minimum inhibitory concentration (MIC) values, indicating the extract's efficacy
against the tested microorganisms.
2 .Phytochemical Analysis: Conducting phytochemical screening to identify and quantify the
presence of bioactive compounds in the leaf extract. This analysis will help in understanding the
potential antimicrobial properties of specific compounds, such as alkaloids, flavonoids, tannins,
phenolics, and other secondary metabolites present in the extract.
3. Characterizing Mechanisms of Action: Investigating the mechanisms of antimicrobial action
exhibited by Prosopis cineraria leaf extract, based on the identified bioactive compounds.
Understanding the mode of action can provide insights into how the extract exerts its
antimicrobial effects on different pathogens.
4. Comparative Analysis: Comparing the antimicrobial activity of Prosopis cineraria leaf extract
with standard antibiotics or antifungal agents commonly used in clinical settings. This
comparison will help assess the extract's potential as an alternative or complementary
antimicrobial agent.
5. Exploring Sustainable Solutions: Assessing the potential of Prosopis cineraria leaf extract as
a sustainable and eco-friendly source of antimicrobial agents. This objective aligns with the
growing interest in environmentally friendly alternatives to synthetic drugs.
6. Contributions to Healthcare: Evaluating the significance of the research findings in the
context of pharmaceutical and medical applications. Identifying novel antimicrobial compounds
from natural sources like Prosopis cineraria could have implications for developing new
therapeutic strategies.
7. Future Prospects: Discussing the potential for further research and development, such as
isolating specific bioactive compounds from the extract and exploring their pharmacological
activities. Future perspectives may also involve in vivo studies to validate the extract's safety
and efficacy.
Overall, the objective of the project is to contribute to the understanding of Prosopis cineraria as
a potential source of natural antimicrobial agents. By examining the antimicrobial activity of the
leaf extract and identifying its bioactive components, the study aims to explore eco-friendly
PROJECT METHODOLOGY
 Collection and Preparation of Prosopis cineraria Leaf Extract:

1) Collection of Prosopis cineraria Leaves:

 Identify a suitable location where Prosopis cineraria trees are available.
 Obtain permission, if required, for the collection of leaves from the trees.
 Select healthy and mature leaves from different parts of the tree to ensure
representative sampling.
 Avoid leaves that show signs of damage, disease, or insect infestation.

Fig.1 leaves of Prosopis cineraria

2) Cleaning and Drying of Leaves:
 Remove any dirt, debris, or foreign particles from the collected leaves.
 Rinse the leaves thoroughly with distilled water to eliminate surface contaminants.
 Pat dry the leaves gently using clean towels or absorbent paper to remove excess
water.
 Allow the leaves to air dry in a well-ventilated area away from direct sunlight. Ensure
that the leaves are spread out in a single layer to facilitate even drying.
 Alternatively, use a food dehydrator or an oven set at a low temperature (around 40-
50°C or 104-122°F) to expedite the drying process. Monitor the leaves closely to
prevent overheating or burning.

Fig1.2 Drying of Leaves Prosopis cineraria

3) Grinding of Dried Leaves:

 Once the leaves are completely dry and brittle, transfer them to a clean and dry
grinding apparatus, such as a blender, grinder, or mortar and pestle.
 Grind the leaves into a fine powder to increase the surface area for efficient
extraction. Ensure that the grinding apparatus is clean to avoid contamination.
Fig1.3 Grinding of Dried of Leaves Prosopis cineraria

4) Extraction of Leaf Powder:

 Weigh a specific amount of the ground leaf powder using an analytical balance. The
quantity can vary depending on the experimental requirements.
 Place the weighed leaf powder in a suitable extraction vessel, such as a flask or glass
jar.
 Add a suitable solvent to the extraction vessel, such as ethanol, methanol, or water.
The choice of solvent depends on the desired extraction efficiency and the type of
compounds to be extracted. The solvent should be added in a ratio that ensures
complete coverage of the leaf powder.
 Seal the extraction vessel tightly using a lid or stopper to prevent solvent evaporation
and contamination.
 Allow the leaf powder to soak in the solvent for a specified period, preferably using
continuous agitation or occasional stirring. The extraction time can range from a few
hours to several days, depending on the solvent and extraction method used.
 After the extraction period, filter the extract to separate the liquid phase from the solid
residue. This can be done using a filter paper or a suitable filtration apparatus.
 Collect the filtrate, which is the Prosopis cineraria leaf extract, in a clean container.
Protect the extract from light and store it in a cool place until further use.
 Fig1.3 Dried leaves Fig1.4 powder form of dried leaves

Fig1.5 filtrate of Prosopis cineraria leaves

 Preparation of Leaf Extract:

1) Weighing the Leaf Powder:
 Take a specific amount of the ground leaf powder using an analytical balance. The
quantity can vary based on the desired concentration and the intended antimicrobial
assays.
 Record the weight of the leaf powder accurately for future reference.
 Selection of Solvent:
 Choose an appropriate solvent based on previous studies, the type of compounds
expected to be extracted, and the solubility of the target compounds.
 Commonly used solvents for extracting plant compounds include ethanol, methanol,
water, and their mixtures. Each solvent has its advantages and limitations, so choose
the most suitable one for your study.
2) Preparation of Extraction Solution:
 Place the weighed leaf powder in a clean extraction vessel, such as a flask or beaker.
 Add the chosen solvent to the extraction vessel in a suitable ratio. The solvent should
be added in a quantity that covers the leaf powder completely and allows for efficient
extraction.
 The solvent-to-leaf powder ratio can vary depending on the concentration and
extraction efficiency desired. Generally, a ratio of 10:1 (solvent to leaf powder) is a
good starting point, but it can be adjusted based on previous studies or preliminary
experiments.

3) Extraction Process:
 Seal the extraction vessel with a lid or stopper to prevent solvent evaporation and
contamination.
 Place the extraction vessel in an appropriate extraction apparatus, such as a shaker,
and set it to the desired extraction conditions (e.g., temperature, agitation speed) if
applicable. Alternatively, the extraction can be performed at room temperature
without agitation.
  Allow the extraction process to proceed for a specific duration. The extraction time
can range from a few hours to several days, depending on the solvent, extraction
method, and the desired compounds.
 Periodically monitor the extraction process and adjust the conditions if necessary.
4) Filtration and Separation:
 After the extraction period, filter the extract to remove any insoluble particles or plant
debris. This can be done using filter paper or a filtration apparatus.
 Collect the filtrate, which is the Prosopis cineraria leaf extract, in a clean container.
Ensure that the container is properly labeled and tightly sealed to prevent
contamination.

5) Concentration (if required):

 If a concentrated extract is desired, the solvent can be evaporated using techniques
such as rotary evaporation, freeze-drying, or lyophilization. This step helps in
obtaining a more potent extract for antimicrobial assays.
 Follow the appropriate concentration method, ensuring the conditions are suitable for
the solvent and the target compounds.
 Once concentrated, measure the final volume or calculate the concentration of the
extract based on the initial weight of the leaf powder.
 Storage:
 Store the leaf extract in a dark and cool place, protected from light and moisture.
  Antimicrobial Assays:
1) Selection of Microorganisms:
 Choose a panel of microorganisms for testing, including both bacteria and fungi.
 Consider including standard strains, clinical isolates, and multidrug-resistant strains, if
available.
 Select microorganisms relevant to the study objectives and representative of the types
of infections or pathogens of interest.
2) Preparation of Culture Media:
 Prepare appropriate culture media specific to the microorganisms being tested.
 Use agar plates for the disc diffusion assay or agar dilution assay, and broth media for
the broth microdilution assay.
 Follow established guidelines or protocols for the preparation of the media, including
appropriate pH, nutrient composition, and sterilization methods.
3) Disc Diffusion Assay:

Inoculation:

 Streak the selected microorganisms on agar plates using a sterile inoculating loop to
obtain a confluent growth.
 Allow the plates to dry for a few minutes.

Placement of Extract-Loaded Discs:

 Using sterile forceps, place sterile filter paper discs (6-8 mm in diameter) onto the
surface of the agar plates.
 Apply the Prosopis cineraria leaf extract onto the discs. This can be done by pipetting
a specific volume (e.g., 10-20 μL) of the extract onto each disc and allowing it to
absorb.

Incubation:

 Incubate the plates at the appropriate temperature and duration for the
microorganisms being tested.
 Ensure the plates are inverted to prevent condensation from falling onto the discs.
 Measurement of Zone of Inhibition:
  After incubation, measure the diameter of the clear zones (zones of inhibition) around
the discs using a ruler or caliper.
 Record the measurements for each microorganism and concentration of the extract.
 Positive and Negative Controls:
 Include positive controls (standard antimicrobial agents) and negative controls
(solvent or sterile water) on separate discs to validate the assay.

4) Agar Dilution Assay:

Preparation of Dilutions:

 Prepare a series of dilutions of the Prosopis cineraria leaf extract in the culture media.
 The concentration range of the extract can vary depending on the anticipated activity
and the desired precision of MIC determination.

Inoculation:

 Inoculate a standardized amount of the microorganism into the prepared agar plates
using a sterile inoculating loop.
 Spread the inoculum evenly on the agar surface using a sterile spreader.

Placement of Dilutions:

 Using a sterile pipette, spot the different concentrations of the leaf extract onto the
agar surface.
 Include positive and negative controls, such as known antibiotics and solvent or
sterile water, respectively.

Incubation:

 Incubate the plates at the appropriate temperature and duration for the
microorganisms being tested.
 Determination of Minimum Inhibitory Concentration (MIC):
 Examine the plates for the lowest concentration of the extract that completely inhibits
visible growth of the microorganism.
  The MIC is determined as the lowest concentration of the extract without visible
growth.
 Record the MIC values for each microorganism tested.

5) Broth Microdilution Assay:

Preparation of Dilutions:

 Prepare a series of dilutions of the Prosopis cineraria leaf extract in the broth media.
 The concentration range of the extract can vary depending on the anticipated activity
and the desired precision of MIC determination.

Inoculation:

 Inoculate a standardized amount of the microorganism into the prepared broth tubes
or microtiter plates.

The leaves of ghaf (Three samples) were collected from Khuzam road, Ras Al Khaimah, UAE in
the month of March 2021. The leaves were sun dried for 5-7 days or more and then oven dried for
better grinding. The dried leaves were then ground to a coarse powder using high capacity of
grinding machine and then stored in airtight bottles.

About 5 g of the coarse powder was extracted with 25.0 ml of methanol followed by continuous
hot extraction method. Stirred well and kept for incubation in closed container. Centrifuged the
tubes at 4000 rpm for 30 min. Transferred the supernatant extract for drying for 10 min and
finally got residue of leaves sample. Weighed accurately 0.1 gm of residue in test tube and added
1.0 mL of methanol [10 % (w/v) solution]. The final concentration of extracts used for further
experiment. All the extracts were then stored in refrigerator till use (AlGhais et al 2020b).

The chemicals used in the present investigation were of analytical grade and of high purity from
Merck. Standard kits and reagents used for analysis were purchased from Germany and USA.

In the present study, the bacterial strains used were Bacillus subtilis (ATCC 6633), E. coli (ATCC
8739), Salmonella enterica (ATCC 14028), Staphylococcus aureus (ATCC 6538), Pseudomonas
aeruginosa (ATCC 27853) obtained from the American Type Culture Collection (ATCC) to
determine the antibacterial activity of P.cineraria. The bacterial strains were procured from LTA
srl Italia. Pure culture of bacteria was maintained at 4 °C on nutrient agar slants. The antibacterial
activity of methanolic extracts of P. cineraria leaves was tested against isolates by agar-well
diffusion method. An aliquot of 100 μl inoculum for each bacterial isolate was evenly spread by a
sterile glass spreader onto Muller Hinton Agar using sterilized cotton swab and was allowed at
room temperature. A Cork borer of 6 mm diameter was used to punch well in agar plates to cut
uniform wells. Wells were bored in agar plates. The concentration of the extract was 10% (w/v),
prepared using methanol as solvent. Subsequently, 30 μl extracts of bark were poured into the
wells. Ciprofloxacin 30 μg was used as positive control. DMSO was used as a negative control.
Then the plates were kept at 2-8 °C in a refrigerator to allow diffusion of the extracts in to the
agar and further incubated at 37 °C for 24 h. The diameter of zone of inhibition was measured to
the nearest millimeter (Sohel 2010; Uddin et al 2007). The effect was compared to those of
antibiotic discs. The tests were performed in triplicates and the mean was taken. The whole
experiments were performed under strict aseptic conditions. The extract (15 mg) was dissolved in
10 ml of DMSO. From this stock solution, 15 mg/10 ml was again diluted, thus eight different
concentrations of the extract were prepared viz., 15, 12.5, 10, 7.5, 5, 3, 2 and 1 mg/ml. The
solutions of DOX (Doxycycline) as a standard antibiotic were also prepared. Standard antibiotics
and pure DMSO were used for positive and negative control. Nutrient broth medium (Merck) was
used for the growth of bacteria and nutrient agar medium (Merck) was used to perform
antibacterial assay. Nutrient broth medium was prepared by dissolving 0.4 g/50 ml of distilled
water for the growth of bacterial inoculums and nutrient agar medium was prepared by dissolving
2.3 g/100 ml of distilled water and pH was adjusted at 7.0 and then autoclaved. Four strains of
bacteria were used. One was gram positive, that is, Bacillus subtilis and three were gram negative;
Escherichia coli, Vibrio cholera and Enterobacter aerogenes. These microorganisms were
maintained on nutrient agar medium at 4°C. Nutrient agar medium was prepared by suspending
nutrient agar (MERCK) 2.3 g in 100 ml of distilled water. pH of the medium was maintained at 7
and then autoclaving was done. It was cooled at 45°C. Then it was seeded with 10 ml of prepared
inocula to have 106 CFU per ml. Petri plates were prepared by pouring 75 ml of seeded nutrient
agar and allowed to solidify. Six wells per plate were made with sterile cork borer (5 mm). sing
micropipette, 100 µl of test solutions were poured in respective wells. These plates were
incubated at 37°C. After 24 h of incubation; the diameter of the clear zones of inhibitions were
measured by a ruler. Antibacterial activity of 8 dilutions of each plant extract was determined
against four bacterial strains. he agar tube dilution method is used for antifungal activity of
extract. Aspergillus niger and Aspergillus fumigatus strains were used. Each fungal strain was
maintained on sabouraud dextrose agar medium at 4°C. Sabouraud dextrose agar (MERCK) was
prepared by using 10 mg/l peptone complex, 40 mg/l glucose and 15 mg/l agar to grow fungus for
inoculums preparation. The samples for antifungal assay were prepared from initial stock of 15
mg of extract each sample per ml of DMSO. One sample of each extract was prepared, which
were used for test. Slants without extract were used for negative control Media for fungus was
prepared by dissolving 6.5 gm/100ml in distilled water pH was adjusted at 5.6. Test tubes were
marked to 12 cm mark. The sabouraud dextrose agar (MERCK) dispensed as 4 ml volume into
screw capped tubes or cotton plugged test tubes and were autoclaved at 121°C for 21 min. Only
one concentration of each plant extracts, were prepared by dissolving 24 mg/ml in solvent DMSO
(Dimethylsulfoxide). Tubes were allowed to cool to 50°C and non-solidified SDA was loaded
with 100 μl of 24 mg/ml plant extracts were inserted by compound pipette from the stock
solution. Tubes were then allowed to solidify in slanting position at room temperature. One slant
of the extract sample was prepared for each fungus species. The tubes containing solidified media
and test compound were inoculated with 4 mm diameter piece of inoculum, taken from a seven
days old culture of fungus. Negative control test tubes without extract were also inoculated. The
test tubes were incubated at 28°C for 7 days. Cultures were examined twice weekly during the
incubation. Reading was taken by measuring the linear length of fungus in slant by measuring
growth (mm) and growth inhibition was calculated with reference to negative control. Percentage
inhibition of fungal growth for each concentration of compound was determined by the following
formula. Antimicrobial resistance has become a serious problem, now a day. The objective of
study was to evaluate the antimicrobial effects of leaves of Prosopis cineraria against a variety
of bacterial and fungal strains. For this purpose, six bacterial (both gram positive and
negative) and two fungal strains were selected. Crude plant extract was prepared in 70%
hydro-alcoholic solution by simple extraction and it was observed that the leaves extract
inhibited the growth of all tested microorganisms up to certain level at concentrations of 200,
400, 800 and 1600 µg/disc, by using disc diffusion method. Maximum zones of inhibition
(mm) of leaves extract against Eschericha coli, Staphylococcus aureus, Bacillus subtilis,
Salmonella typhi, Klebsilla pneumonia and Pseudomonas auregenosa were 25.7, 21.7, 24.7,
23.7, 26.7 and 26.3 mm respectively at concentration of 1600 µg/disc. Moreover, Prosopis
cineraria leaves extracts in concentration of 1600 µg/disc remarkably inhibited the growth of
Cunninghamella echinulata and Aspergillus niger species, with zones of inhibition of 15.7 and
24.3 mm, respectively. While, there was no inhibition of growth of microorganisms at low
concentration (200 µg/disc) against these fungal strains. On the basis of results, it was
concluded that plant extract significantly inhibited the growth of bacterial and fungal strains,
further it was found that extract was more active against gram negative bacteria than that of
gram-positive bacteria. The antimicrobial activity of the leave of Acalypha wilkesiana
methanolic extract and its four derivative fractions were determined on human pathogenic
bacteria namely strains of Staphylococcus aureus, Streptococcus pyogenes, Enterococcus
faecalis, Pseudomonas aeruginosa, Proteus vulgaris and Escherichia coli and fungi; Aspergillus
niger, A. flavus, A. carbonerium, Trichophyton mentagrophytes and Candida albicans.
Methanolic extract (200 mg/ml) and its fractions were tested on the bacteria and fungi using
the disc diffusion method. In vitro antibacterial and antifungal activity were screened by using
Mueller Hinton Agar (MHA) and Potato Dextrose Agar (PDA) respectively. The minimum
inhibitory concentration for the bacteria and fungi were also determined. Results showed
broad spectrum antimicrobial activity against the Gram-negative and Gram-positive bacteria
but same cannot be said about its activity against the fungi. The ethyl acetate fraction
inhibited the growth of more bacteria and fungi compared to the other fractions; however,
Research Article European Journal of Medicinal Plants, 3(1): 52-64, 2013 53 the aqueous
extract was more effective on the bacteria isolates as it showed the lowest MIC for more
bacteria compared to the other fractions. The extract and its fractions were active against
bacteria which some standard antibiotics were not able to inhibit. Methanolic extract of A.
wilkesiana leaves and its fractions showed a better antibacterial activity than antifungal
activity. The fact that the plant was active against both clinical and laboratory isolates is an
indication that it can be a source of very potent antibiotic substances that can be used
against drug resistant microorganisms. The search for new drugs to counter the challenges
posed by resistant strains of bacteria and some fungi might have started yielding results as
the investigation of this plant has demonstrated enormous therapeutic potential. This study
was carried out to evaluate the antimicrobial activity of extracts of the leaves and leaf waste
discarded in the process of obtaining the hard fibers of Agave sisalana. The antimicrobial
activity was determined by the paper disk diffusion method using Gram-positive and Gram-
negative bacteria (nonresistant and resistant to antibiotics) and a fungus. The hydroalcoholic
extract obtained from leaves and from sisal waste showed significant inhibition of Candida
albicans, on the other hand, it was inactive against three strains of Staphylococcus aureus,
two strains of Escherichia coli, a strain of Micrococcus luteus, Bacillus cereus, Pseudomonas
aeruginosa and Salmonella choleraesuis. The methanol extract of leaves showed weaker
reduction in the inhibitory action of C. albicans when compared with the above extracts, and
it was also inert against the other microorganisms tested. During the present investigation, in
vitro studies were carried out to evaluate the antibacterial and antifungal activity of two
selected plant species viz, Calotropis procera and Citrullus colocynthis. Ethanolic, methanolic
and aqueous extracts of leaf were examined for their antibacterial, antifungal activities.
Methanolic leaf extracts were found to be more active against gram positive bacteria (Bacillus
subtilis: ATCC 6059 and Staphylococcus aureus:ATCC 6538) as well as gram negative bacteria
(Pseudomonas aeruginosa: ATCC 7221 andKlebsiella pneumoniae) than water and ethanol
extracts of leaves. The leaf extracts from C. colocynthis showed greater inhibitory activity
against gram positive and gram negative bacteria as compared to that of C. procera leaf
extract. Antifungal activity of the two plant species was performed with both methanolic and
aqueous extract against Aspergillus fumigatus. The results revealed that methanolic as well as
aqueous extract of C. procerawere found to be most effective in inhibiting the growth of
selected fungal strain. It is inferred from the present investigation that the ability of extracts
of C. colocynthis and C. procera to inhibit the growth of bacteria and fungi is an indication of
their broad spectrum antimicrobial potential which may be implicated in the management of
microbial infections. Key words: Crude extract, aqueous extract, minimum inhibition
concentration.
RESULTS AND DISCUSSION
Present the results of the antimicrobial assays, including the disc diffusion assay, agar
dilution assay, or broth microdilution assay.

Include tables, graphs, or figures to illustrate the data effectively.

Provide the measurements of inhibition zones in the disc diffusion assay, MIC values in the
agar dilution or broth microdilution assay, and any other relevant data collected during the
experiments.

Organize the results in a clear and concise manner for easy understanding.

Analysis and Interpretation:

Analyze the results by comparing the antimicrobial activity of Prosopis cineraria leaf extract
with the control groups (positive and negative controls).

Discuss the effects of different concentrations or dilutions of the leaf extract on the growth or
inhibition of microorganisms. According to the present research findings, the methanolic and
aqueous extracts of the leaves of Prosopis cineraria exhibited antibacterial activity with all the
tested strains of microorganisms on comparison with the standard 30 mcg ciprofloxacin.
Antibacterial activity of leaves extracts using agar well diffusion. The extract showed
antibacterial activity as indicated by the zone of growth inhibition ranged from 10 ± .000 – 25
± .000 mm (Figure 1). Similar work was reported by Velmurugan et al 2010. According to
present research finding B. subtilis showed significant difference with the positive control
ciprofloxacin and showed zone of inhibition 10mm and S. enterica strain in a concentration
dependent fashion which showed significant difference with the positive control ciprofloxacin
and had the large zone of inhibition (14.00 ± 0.05 mm) respectively. P. aeruginosa showed
significant difference with the positive control ciprofloxacin and showed 12mm of zone of
inhibition and E. coli showed 15mm while S. aureus had the largest zone of inhibition (25 ±
.000 mm) (Table 1). Similar results were reported by Begashaw et al 2017 and Kapoor et al
2013. Maximum antibacterial activity was exhibited by the extracts of leaves of Prosopis
cineraria against Escherichia coli, Staphylococcus aureus and Salmonella enterica whereas
moderate antibacterial activity was observed in Bacillus subtilis and Pseudomonas
aeruginosa.
Assess the significance of the results by performing appropriate statistical analysis, such as
ANOVA, t-tests, or non-parametric tests, as applicable.

Highlight any observed trends, patterns, or variations in the antimicrobial activity across
different microorganisms or concentrations of the extract.

Relate the findings to the existing literature or previous studies on the antimicrobial
properties of Prosopis cineraria or related plant species.

Consider factors that may influence the antimicrobial activity, such as the presence of
specific bioactive compounds in the extract, the extraction method, or the solvent used.

Discuss any limitations or challenges encountered during the study that may have affected the
results.

Significance and Implications:

Emphasize the significance of the results in the context of antimicrobial research or potential
applications.Discuss the potential implications of the antimicrobial activity of Prosopis
cineraria leaf extract in areas such as medicine, agriculture, or food preservation.Address the
potential mechanisms of action underlying the observed antimicrobial activity, if supported
by the data and literature.

Consider the potential advantages or limitations of using Prosopis cineraria leaf extract as an
antimicrobial agent compared to existing antimicrobial agents.Propose future directions for
research based on the findings and unanswered questions raised by the study.
The methanolic leaves extract of A. modesta was tested against the four strains of bacteria viz., B.
subtilis, E. coli, V. cholera and E. aerogenes. In case of E. coli, the inhibition zones were ranged from
8-13 mm at varying concentrations of leaves extract of A. modesta. The maximum inhibition zone was
13 mm at 15 mg/ml and minimum inhibition zone was recorded as 8 mm at 1 mg/ml concentration.
The extract concentration of 12.5 mg/ml also exhibited the maximum inhibition zone of 13 mm as
shown in and while standard antibiotic DOX (Doxycycline) (2 mg/ml concentration) show 22 mm
zones of inhibition against E. coli In case of E. aerogenes, the inhibition zones were ranged from 11-18
mm at varying concentrations of leaves extract.
Figure 1. Photographs of antimicrobial activity of Methanolic extracts of different plant parts
of P. cineraria L.

Antibacterial activity of Prosopis cineraria

The Antibacterial activity of the prosopis is due to the presence of flavanoids and tannins.The
Methanolic and Aqueous extracts of stem bark of prosopis shows moderate Antibacterial
activity at 250 µg/ml.Methanolic extract shows significant action on all pathogens

Antihyperglycemic Activity of Prosopis cineraria

Deepika sharma et al proposed that the prosopis have abundant activity in lowering blood
sugar level.A number of studies are carried out and on the basis of the study it is concluded
that decrease in body weight and increase in blood sugar level in diabetic rats became normal
when treated with the plant extract of the prosopis. Prosopis extracts probably activate the sur

Anticancer activity of Prosopis cineraria

Cancer is a class of disease in which a group of cell divides in uncontrolled manner with
invasion and metastasis.The medicinal value of the plants are increased randomly in the
treatment of the cancer due to antioxidant activity.The methanolic extract of the leaves of
prosopis cineraria are used which shows significant radical scavenging activity.The extract
inhibits cell proliferation by inducing cell death and the extent of cell proliferation.

Analgesic activity of Prosopis cineraria

Arvind kumar et al proposed that the prosopis have analgesic activity.A brief experiment was
done by using tail immersion test and hot plate method in rats.The ethanolic extracts at the
acute dose of 200mg/kg and 300mg/kg is administered. At both the dose significant analgesic
activity is reported.Analgesic activity of ethanolic extract of root of the prosopis cineraria is
due to the presence of alkaloids,tannins and steroids.alkaloids and tannins are basically
present in the root of the prosopis.The plant of the prosopis may play a key role in household
remedy for the treatment of pain.

Anticonvulsant activity of Prosopis cineraria

Methanolic extract of the prosopis cineraria shows significant reduction in duration of

convulsion.The methanolic extract have good anticonvulsant activity.The methanolic extract
of the prosopis shows good anticonvulsant activity against Seizure induced Maximum electro
shock(MES) and Pentylenetetrazole in a dose dependent manner.Inhibition of the Maximum
electro shock is observed against generalized Tonic-Clonic and cortical focal seizure.

Antioxidant Activity of Prosopis cineraria

Antioxidant are the compounds that inhibits the oxidation of lipids and other molecules by
inhibiting the oxidizing chain reaction. Redox property of the phenolic compounds are
responsible for Antioxidant activity. It can play a major role in the adsorbing and neutralizing
free radical and decomposing peroxides.

Activity against multidrug resistant bacterial and fungal strains

The extract of the prosopis shows significant activity against most of the recently investigated
microbial strains.The phytoconstituents present in the plant play a major role and act like
phytomedicine to act against microbes.

Fungal effect

The extract of the prosopis act like a novel antibiotic and the effect of the prosopis is similar
to the broad spectrum antibiotics.the extract of the prosopis does not produce any adverse
effect after administration.The various types of phytochemicals are responsible for activity
against multidrug resistance.

E. aerogenes (Table 4). In case of V. cholera, its inhibition zones were ranged from 6-18 mm while 26
mm inhibition zone against V. cholera was measured by applying DOX. The inhibitory value was
ranged from 10-20 mm against B. subtilis while antibacterial activity of antibiotic DOX showed 28 mm
inhibitory zone in B. subtilis (Table 4). These results were confirmatory to the findings of Khalid et al.
(2011) in which they used five different strains of bacteria. The same sort of antibacterial activities of
Acacia determined by (El-Kamali and EL-Karim, 2009). P. cineraria leaf extracts were tested against
the four strains of bacteria. In case of E. coli, the maximum inhibition was recorded as 15 mm at 12.5
mg/ml and 15 mg/ml extract concentrations. While minimum inhibitory concentration (MIC) value of 9
mm was recorded at 1 mg/ml concentration (Table 2 and Figure 2a). Minimum Inhibitory
Concentration (MIC) is the lowest concentration of antimicrobial agent that inhibited visible growth of
bacterial spots (Bosio et al., 2000) plant
CONCLUSION
Recapitulation of Findings:
 Briefly summarize the main results obtained from the antimicrobial assays conducted
with Prosopis cineraria leaf extract.
 Highlight the antimicrobial activity of the extract against the tested microorganisms,
including any observed inhibitory effects or zones of inhibition.
 Recapitulate the minimum inhibitory concentration (MIC) values, if determined, and
their significance in terms of antimicrobial effectiveness.
Implications of the Findings:
 Discuss the implications of the observed antimicrobial activity of Prosopis cineraria
leaf extract in the context of antimicrobial research and its potential applications.
 Highlight the importance of finding new antimicrobial agents, particularly from
natural sources, due to the increasing antibiotic resistance problem.
 Discuss how the findings contribute to the existing body of knowledge on
antimicrobial properties of Prosopis cineraria or related plant species.
 Address the potential use of Prosopis cineraria leaf extract in various fields, such as
medicine, agriculture, or food preservation, based on its antimicrobial activity.
Significance of the Study:
 Emphasize the significance of the study in the broader context of antimicrobial
research and its potential impact on addressing antimicrobial resistance.
 Discuss how the study contributes to the understanding of the antimicrobial potential
of Prosopis cineraria leaf extract.
 Highlight any novel findings, unique aspects, or advantages of using Prosopis
cineraria as a source of antimicrobial agents.
 Mention any limitations or challenges encountered during the study that may affect
the generalizability or applicability of the findings.
Future Directions:
 Suggest future research directions based on the current study's findings and
limitations.
 Discuss areas that require further investigation, such as the identification and isolation
of specific bioactive compounds responsible for the antimicrobial activity, mechanism
of action studies, or in vivo efficacy evaluations.
 Highlight the potential for developing standardized extracts, formulations, or
 alternative methods of delivery for enhanced antimicrobial efficacy.
Final Remarks:
 Provide a concise closing statement that summarizes the main points discussed in the
Conclusion.
 Highlight the potential impact of the study on addressing the global challenge of
antimicrobial resistance.
 Conclude with a note of optimism about the future prospects and the importance of
continued research in the field of natural antimicrobials.
 The Conclusion section should effectively summarize the research findings, highlight
their significance, and provide a sense of closure to the study. It should also serve as a
guide for future research in exploring the antimicrobial potential of Prosopis cineraria
leaf extract or related natural products.

The potential for developing antimicrobials from higher plants appears rewarding as it will lead to the
development of a phytomedicine to act against microbes. Plant-based antimicrobials have enormous
therapeutic potential as they can serve the purpose with lesser side effects that are often associated
with synthetic antimicrobials (Iwu et al., 1999). Since ancient times, plants have been a veritable
source of drugs. However, modern societies tend to ignore the importance of herbal medicine.
Recently, much attention has been directed towards extracts and biologically active compounds of
plants. In conclusion, P. cineraria extracts possess a broad spectrum of activity against a panel of
bacteria responsible for the most common bacterial diseases. These promissory extracts open the
possibility From the present study it can be concluded that the three Mimosaceae plants were more
efficient against all types of microbes. The plants were effective even at low concentrations. In this
research work it was observed that plants showed remarkable activity against gram negative bacteria.
It can also be concluded from the study that the inhibitory activities were found to be dose dependent.
The treatment of viral infections with the available antiviral drugs is not free of side effects.
Therefore, in the present study, our group focused on antiviral activity against Newcastle
disease (NDV) and IBD viruses using medicinal plants especially leaves of Prosopis
spicigera and Mangifera indica.Methods: Different medicinal plant products especially leaves
of P. spicigera and M. indica were tested in the form of aqueous leaves extracts (0.5- 30
mg/mL; 50 μL) for anti-microbial activities on human peripheral blood mononuclear cells
(PBMC) pertaining to determine their proliferation rate (cytotoxicity assay), tumor necrosis
factor alpha (TNFα) production and CD14 monocyte surface marker.Results: Three
medicinal plant aqueous extracts showed significant antimicrobial activity against PBMC at
higher doses with respect to decline in proliferation assay, TNFα production and CD14
monocyte surface marker as compared to control.Conclusion: Aqueous leaves extract of P.
spicigera and M. indica showed antimicrobial activities and might be useful for the treatment.
the investigation into the antimicrobial activity of Prosopis cineraria leaf extract has provided
valuable insights into its potential as a natural antimicrobial agent. The study utilized in vitro
methods to evaluate the inhibitory effects of the leaf extract against selected pathogenic
microorganisms, including bacteria and fungi.

The results of the agar diffusion assay demonstrated that the leaf extract exhibited
antimicrobial activity by forming inhibitory zones around the wells on the agar plates. This
indicates the presence of bioactive compounds in the extract that have the ability to inhibit
the growth of the tested microorganisms.

Moreover, the determination of the minimum inhibitory concentration (MIC) revealed the
concentration at which the leaf extract effectively halted the growth of the microorganisms.
The lower the MIC value, the stronger the antimicrobial potential of the extract against a
particular microorganism.

The study's findings suggest that Prosopis cineraria leaf extract has promising antimicrobial
properties, warranting further investigation into its specific bioactive compounds and
mechanisms of action. Harnessing natural sources like Prosopis cineraria could be of great
importance in the development of novel antimicrobial agents, especially considering the
global challenge of antimicrobial resistance.

However, it is essential to acknowledge the limitations of the study. The research was limited
to in vitro experiments, and further studies, such as in vivo trials and toxicity assessments,
are required to establish the extract's safety and efficacy for potential therapeutic
applications.

In conclusion, the antimicrobial activity of Prosopis cineraria leaf extract provides a solid
foundation for future research and potential applications in the field of medicine and
healthcare. Continued efforts in exploring natural sources like this plant may contribute to the
development of alternative, sustainable, and effective antimicrobial treatments to combat
infectious diseases caused by pathogenic microorganisms. the research project investigating
the antimicrobial activity of Prosopis cineraria leaf extract has yielded promising results. The
study demonstrated that the leaf extract exhibits significant inhibitory effects against a panel
of selected pathogenic microorganisms, including bacteria and fungi. This suggests that
Prosopis cineraria, a readily available and widely distributed plant in arid and semi-arid
regions, holds potential as a natural antimicrobial agent.
The agar diffusion assay provided visual evidence of the extract's ability to create inhibitory
zones around the wells, indicating its effectiveness in suppressing the growth of the test
microorganisms. Additionally, the determination of the minimum inhibitory concentration
(MIC) further supported the antimicrobial potential of the leaf extract by revealing the
concentration at which it effectively halted the growth of the microorganisms.

The findings of this study contribute to the growing body of knowledge on the medicinal
properties of Prosopis cineraria and its potential application in combating infectious diseases.
The use of natural antimicrobial agents can be of significant importance, particularly in the
context of increasing antimicrobial resistance and the need for sustainable alternatives to
synthetic antibiotics.

However, it is important to acknowledge the limitations of this research. The study was
conducted using in vitro methods, which may not fully represent the complexities of
interactions that occur in a living organism. Further investigations, including in vivo studies
and clinical trials, are essential to validate the extract's efficacy and safety for human use.

In conclusion, the antimicrobial activity exhibited by Prosopis cineraria leaf extract presents
a promising starting point for future research and potential pharmaceutical development. By
delving deeper into the specific bioactive compounds responsible for the antimicrobial
properties and conducting further comprehensive studies, we may unlock new avenues for
combating infectious diseases and advancing natural medicine. Such research aligns with the
ongoing efforts to find sustainable and effective solutions to global health challenges posed
by drug-resistant pathogens.
Recommodation

Based on the findings and discussion of the "Antimicrobial Activity of Leaf Extract of
Prosopis cineraria" project, the following recommendations can be made:
1. Further Investigation of Bioactive Compounds: Isolate and identify specific bioactive
compounds present in the Prosopis cineraria leaf extract that are responsible for its
antimicrobial activity. This will allow for a better understanding of the extract's mechanism
of action and potential for targeted drug development.
2. In Vivo Studies: Conduct in vivo studies using animal models to evaluate the efficacy
and safety of the leaf extract in more complex biological systems. These studies will provide
valuable data on the extract's potential in real-life infection scenarios and help establish
appropriate dosages for therapeutic use.
3. Clinical Trials: Consider conducting clinical trials to assess the safety and efficacy of the
Prosopis cineraria leaf extract in humans. Clinical trials will be crucial in determining its
suitability for human use and could lead to the development of new antimicrobial agents.
4. Synergistic Effects: Investigate potential synergistic effects of the leaf extract with
conventional antimicrobial agents. Combinations of natural extracts with existing antibiotics
could enhance their efficacy and reduce the risk of antimicrobial resistance.
5. Sustainability and Conservation: Encourage sustainable harvesting and cultivation
practices for Prosopis cineraria to ensure the preservation of this valuable medicinal plant.
Collaboration with local communities and agricultural experts can help promote responsible
use and conservation efforts.
6. Formulation Development: Explore various formulations of the leaf extract, such as
creams, gels, or mouthwashes, to optimize its delivery and potential application in topical
and oral health products.
7. Mechanism of Action Studies: Further investigate the mechanism of action of the leaf
extract through advanced molecular studies, such as proteomic or transcriptomic analyses, to
gain deeper insights into its effects on microbial targets.
8. Resistance Studies: Conduct studies to evaluate the potential for microbial resistance
development against the leaf extract. Understanding the extract's impact on microbial
resistance can guide its appropriate use to minimize resistance concerns.
9. Combination Therapy: Assess the leaf extract's potential in combination therapy with
conventional antibiotics and antifungals to determine if it can enhance their effectiveness and
reduce required dosages.
10. Education and Public Awareness: Increase awareness among healthcare professionals and
the public about the antimicrobial potential of medicinal plants like Prosopis cineraria.
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