Open Access

Ethnomedicines and anti-parasitic activities of Pakistani medicinal plants against Plasmodia and Leishmania parasites

  • Akash Tariq1, 2Email author,
  • Muhammad Adnan2,
  • Rahila Amber3,
  • Kaiwen Pan1,
  • Sakina Mussarat2 and
  • Zabta Khan Shinwari4
Annals of Clinical Microbiology and Antimicrobials201615:52

https://doi.org/10.1186/s12941-016-0170-0

Received: 29 June 2016

Accepted: 13 September 2016

Published: 20 September 2016

Abstract

Background

Leishmaniasis and malaria are the two most common parasitic diseases and responsible for large number of deaths per year particularly in developing countries like Pakistan. Majority of Pakistan population rely on medicinal plants due to their low socio-economic status. The present review was designed to gather utmost fragmented published data on traditionally used medicinal plants against leishmaniasis and malaria in Pakistan and their scientific validation.

Methods

Pub Med, Google Scholar, Web of Science, ISI Web of knowledge and Flora of Pakistan were searched for the collection of data on ethnomedicinal plants. Total 89 articles were reviewed for present study which was mostly published in English. We selected only those articles in which complete information was given regarding traditional uses of medicinal plants in Pakistan.

Results

Total of 56 plants (malaria 33, leishmaniasis 23) was found to be used traditionally against reported parasites. Leaves were the most focused plant part both in traditional use and in in vitro screening against both parasites. Most extensively used plant families against Leishmaniasis and Malaria were Lamiaceae and Asteraceae respectively. Out of 56 documented plants only 15 plants (Plasmodia 4, Leishmania 11) were assessed in vitro against these parasites. Mostly crude and ethanolic plant extracts were checked against Leishmania and Plasmodia respectively and showed good inhibition zone. Four pure compounds like artemisinin, physalins and sitosterol extracted from different plants proved their efficacy against these parasites.

Conclusions

Present review provides the efficacy and reliability of ethnomedicinal practices and also invites the attention of chemists, pharmacologist and pharmacist to scientifically validate unexplored plants that could lead toward the development of novel anti-malarial and anti-leishmanial drugs.

Keywords

Ethnomedicines Malaria Leishmaniasis Phytochemicals In vitro activities

Background

Leishmaniasis and malaria are two most common parasitic diseases and infects a large number of human populations worldwide. Leishmaniasis is endemic disease of almost 88 countries in which about 350 million peoples are at risk of infection [1]. Malaria is a major public health problem throughout the world and causes one million deaths per year particularly in developing countries [2].

Leishmaniasis is caused by an obligate intracellular protozoan parasite of genus Leishmania while transmitted to humans and other animals by many species of phlebotomus sand flies [3, 4]. The main causative agents for leishmaniasis are Leishmania tropica, Leishmania major, Leishmania aethiopica, Leishmania donovani and Leishmania infantum. The four main clinical types of leishmaniasis are cutaneous leishmaniasis, mucocutaneous leishmaniasis, diffuse cutaneous leishmaniasis and visceral leishmaniasis. Among these visceral leishmaniasis is very fatal if left untreated. About 90 % cases of visceral leishmaniasis are reported from many countries like Brazil, Bangladesh, Sudan, India and Nepal [5, 6]. Leishmaniasis is a complex group of diseases produces various symptoms in host depending upon parasite’s type [7]. Commonly used allopathic drugs against leishmaniasis are pentavalent antimonials like sodium stibogluconate and meglumine antimoniate etc. These allopathic drugs are mostly unaffordable to the local people and are also not safe due to their toxicity on living system. Many drugs need long term administration to recover from the disease and show side effects depending on the patient’s reaction to medicine [8].

Malaria is caused by an intra-erhytrocytic protozoan parasite of the genus Plasmodium and transmitted by female anopheles mosquito [9]. The four main species of Plasmodia which infects humans are Plasmodium falciparum, Plasmodium vivax, Plasmodium knowlesi, Plasmodium malariae, and Plasmodium ovale. Globally the most important species is P. falciparum causing severe and potentially fatal malaria [2, 10]. For the treatment of malaria many drugs like chloroquine, halofantrine, pyrimethamine, mefloquine, quinine and artemisinin are used [2, 4]. Many problems like resistance of the parasites to drugs, lack of effective vaccines, resistance of mosquito vectors to insecticides and socioeconomic problems rendering treatment of malaria through chemotherapy ineffective [11, 12].

Leishmaniasis and malaria has become a particular problem in the rural areas of Pakistan of all the provinces [13]. Approximately 66 % people of Pakistan live in rural areas [14] and majority of the rural population in Pakistan is poor and cannot afford such expensive drugs for the treatment of leishmaniasis and malaria [15, 16]. Mostly in rural areas peoples depend on medicinal plants for the treatment of various diseases particularly leishmaniasis and malaria [17]. Traditional medicines are extensively using in Pakistan due to easily affordability and efficacy against various diseases [18, 19].

The present review was designed to gather utmost fragmented published literature on anti-malarial and anti-leishmanial plants used by local people in Pakistan. This review will also provide information on in vitro screening and phytochemical investigation of documented plants against these parasites. Future outcomes of this review are to provide evidences regarding the efficacy and reliability of ethnomedicines against Leishmania and Plasmodia parasites, identify scientific gaps present in current knowledge and to recommend future research areas for the development of effective anti-malarial and anti-leishmanial drugs with fewer side effects.

Methods

This review paper was designed by collecting and consulting large number of mostly published literature on medicinal plants used to treat leishmaniasis and malaria in Pakistan. Pub Med, Google Scholar, Web of Science, ISI Web of Knowledge and Flora of Pakistan were searched for the collection of data on ethnomedicinal plants. Plant list and Tropicos were searched for the corrections of plant scientific names, publication authors, synonyms and families. Different search indicators like ethnomedicinal plants used against leishmaniasis and malaria, in vitro activity of different medicinal flora of Pakistan, epidemiology of leishmaniasis and malaria in world, prevalence of leishmaniasis and malaria in Pakistan, drug resistance potential of Leishmania and Plasmodia parasites were used for the collection of data from the database. Total 89 articles were reviewed for present study which was mostly published in English. We selected only those articles in which complete information was given regarding traditional use of medicinal plants in Pakistan. In-vitro activity of those plants has been mentioned which were checked against Leishmania and plasmodia parasites. On the bases of selected data from literature, three tables were formulated using Microsoft Excel 2007 and Microsoft Word 2007. Tables 1 and 2 were formulated on medicinal plants used to treat leishmaniasis and malaria in Pakistan. These tables contains plant name, family, local name, part used, study area and phytochemistry. Table 3 was formulated on anti-parasitic activity of medicinal plants against L. tropica, L. major and P. falciparum. Concentration of plant’s extract (µg/ml) and their inhibition (%) against parasites were also mentioned. Pure compounds isolated from ethnomedicinal plants and assessed against these parasites have also been mentioned in this review article. Chemical structures of compounds were drawn using ChemDraw software and shown in (Figs. 1, 2 and 3) (CambridgeSoft\ ChemOffice2004\ChemDraw).
Table 1

Medicinal plants used to treat leishmaniasis in Pakistan

S. no

Botanical name/family name

Common and or local/name (s)

Part used

Area

Phytochemistry

Citation

1

Aloe vera (L.) Burm.f./Xanthorrhoeaceae(= Aloe barbadense Mill./Liliaceae)

Kuwargandal, Aloe vera

Leaves

Dera Ismail Khan

Sitosterol

[61]

Kohat

[25, 57]

2

Asparagus gracilis L./Asparagaceae

Shagandal

Aerial parts

Islamabad

Glycosides, tannins, saponins

[22]

3

Asparagus asiaticus L./Berberidaceae (= Berberis baluchistanica Ahrendt.)

Zarch

Roots

Kalat

Alkaloids, flavonoids, saponins, diterpenes, phenols

[62]

4

Trachyspermum ammi (L.) Sprague/Apiaceae (= Carumcopticum L./Umbelliferae)

Ajwain

Whole plant

Quetta

NA

[48]

5

Citrullus colocynthis L. Schrad/Cucurbitaceae

Bitter apple, Kortuma

Fruits

Nushki

Ursolic acid, cucurbitacin E 2-0-β-D-glucopyranoside and cucurbitacin I 2-0-β-D-glucopyranoside, alkaloids, flavonoids, saponins, tannins, terpenoids, diterpenes, coumarins

[35]

6

Juniperus M.Bieb./Cupressaceae

Juniper

Fresh berries

Ziarat

Alkaloids, flavonoids, saponins, diterpenes, phenols

[37]

7

Jurinea dolomiaea Boiss./Compositae

Nazar zela

Roots

Kohistan

Alkaloids, flavonoids, saponins, terpenoids, phenols

[22]

8

Melia azedarach L./Meliaceae

Neem, Chinaberry tree, Persian lilac

Green fruits

Islamabad

Phenols

[21]

9

Nepeta praetervisa Rech.f./Lamiaceae

Simsok

Leaves

Kalat

Carbohydrate, tannins, phenols, alkaloids, flavonoids, diterpenes, quinones, cardiac glycosides, terpenoids, triterpenoids, coumarins

[63]

10

Onosma griffithii Vatke./Boraginaceae

Golden drop

Whole Plant

Malakand

NA

[64]

11

Perotis hordeiformisNees ex Hook. &Arn./Poaceae

Kikuyu grass

Leaves

Soorab

Alkaloids, flavonoids, saponins, diterpenes, phenols

[65]

12

Physalis minima L./Solanaceae

Pygmy Ground cherry, Gooseberry

Whole plant

Karachi

Physalins

[66]

13

Rhazya stricta Decne./Apocynaceae

Aizwarg

Leaves

Nushki

Alkaloids, flavonoids, saponins, diterpenes, phenols

[34]

14

Salvia bucharica Popov./Lamiaceae

Sage, Gul-e-Kakar

Leaves

Quetta

NA

[67]

15

Sarcococca wallichii Stapf/Buxaceae (= Sarcococca coriacea Mull. Arg.)

NA

Roots

Karachi

Steroidal alkaloids

[68]

16

Sarcococca hookeriana Baill./Buxaceae

Sweet box

Whole plant

Karachi

Steroidal alkaloids

[66]

17

Sida cordata L. (Burm.f.) Borss. Waalk./Malvaceae

Simak

Whole plant

Islamabad

Phenols, saponins, flavonoids

[22]

18

Stellaria media L. Vill./Caryophyllaceae

Gander

Whole plant

Islamabad

Glycosides, flavonoids, phenols, saponins, terpenoids

[22]

19

Swertia chirata Roxb ex./Gentianaceae

Chirata

Seeds

D. I. Khan

Amarogentin, amaroswerin, sweroside

[69]

20

Tamarix aphylla (L.) H.Karst./Tamaricaceae

Ghaz, Tamarisk, Salt cedar

Barks

Kohat

NA

[25]

21

Thuspeinanta brahuica (Boiss.) Briq./Lamiaceae

NA

Leaves

Kalat

Alkaloids, flavonoids, saponins, phenols, diterpenes

[70]

22

Tylophora hirsute Wight/Apocynaceae (= Asclepiadaceae)

Damvel

Aerial parts

Malakand

NA

[71]

NA indicates data not available

Table 2

Medicinal plants used to treat malaria in Pakistan

S. no

Botanical name/family name

Common and or local name (s)

Area

Part used

Phytochemistry

Citation

1

Acacia nilotica L. (Delile)/Leguminosae (= Fabaceae)

Kikar

Mardan

Leaves

Terpenoids

[72]

2

Ajuga integrifolia Buch.-Ham./Lamiaceae (= Ajugabracteosa Wall ex Benth./Labiatae)

Rati buti

Maradori valley

Leaves

NA

[73]

3

Allium cepa L./Amaryllidaceae (= Liliaceae)

Piaz

Bannu

Bulb

NA

[74]

4

Artemisia annua L./Compositae (= Asteraceae)

Afsantin jari, Sweet Wormwood

Northern areas

Whole plant

Artemisinin

[75]

Maradori valley

Root

[75]

5

Artemisia japonica Thunb./Compositae (= Asteraceae)

Barmar, Basna Tashang

Northern areas

Whole plant

Artemisinin

[75]

6

Artemisia maritime L./Compositae (= Asteraceae)

Tarkh, Zoon, Rooner

Northern areas

Whole plant

Artemisinin

[75]

7

Artemisia scoparia Waldst. and Kitam./Compositae (= Asteraceae)

Lungi booti

Bhimber

Flowering shoots

Artemisinin

[76]

8

Azadirachta indica A.Juss./Meliaceae

Neem

D. I. Khan

Seeds, Leaves

Limonoid (gedunin)

[73]

9

Bupleurum longicaule Wall.ex DC./Apiaceae

Proshi

Maradori valley

Root

NA

[73]

10

Calotropis procera (Aiton) Dryand./Apocynaceae (= Asclepiadaceae)

Sodom apple, Mudar, Milk weed

Cholistan desert

Root

Alkaloids, Flavonoids, Nitrogen, Crude protein, Crude fiber, Soluble phosphates

[77]

Karachi

[16]

11

Capparis spinosa L./Capparidaceae

Kaveer

Chitral

Flowers

NA

[78]

12

Trachyspermum ammi (L.) Sprague/Apiaceae (= CarumcopticumL./Umbelliferae)

Ajwain

Quetta

Whole plant

NA

[48]

13

Datura stramoniumL./Solanaceae

Jimson weed

Faisalabad

Leaves

Alkaloids, flavonoids, saponins, glycosides, tannic acid, vitamin C, steroids

[79]

14

Dodonaea viscosa (L.) Jacq./Sapindaceae

Ghwarasky

Allai valley

Seeds

NA

[80]

15

Enicostemma hyssopifolium Willd./

Gentianaceae

Chhota Chirayata, Nagajihva

Karachi

Whole plant

NA

[16]

16

Eucalyptus camaldulensis Dehnh./Myrtaceae

River red gum

Karachi

Leaves, stem

NA

[16]

17

Fagonia creticaL./Zygophyllaceae

Azghakey

Mardan

Leaves

Terpenoids

[72]

18

Helianthus annuus L./Compositae (= Asteraceae)

Maera stargay gul, Sunflower

Bannu

Leaves

NA

[74]

19

Melia azedarach L./Meliaceae

Neem, Chinaberry

Tree

Islamabad

Green fruits

Phenols

[19]

20

Moringa oleifera Lam./Moringaceae

Sajna, Marango, Moonja

Faisalabad

Whole plant

NA

[81]

21

Nerium oleander L./Apocynaceae

Adelfa, Rose bay

Faisalabad

Leaves

Alkaloids

[79]

22

Origanum majorana L./Lamiaceae

Sweet marjoram

Faisalabad

Aerial parts

NA

[36]

23

Origanum vulgare L./Lamiaceae

Satar, Pot marjoram

Faisalabad

Aerial parts

NA

[36]

24

Peganum harmala L./Nitrariaceae (= Zygophyllaceae)

Harmal

Northern areas

Seeds

β-carboline alkaloid (isoharmine), harmaline, harmine

[82]

25

Polygonatum verticillatum (L.) All./Asparagaceae

Worlds Solomon’s seal

Swat

Aerial parts

α-Bulnesene, Linalyl, acetate, eicosadienoic, docosane, pentacosane, piperitone

[83]

26

Psidium guajava L./Myrtaceae

Amrood

Mardan

Leaves

Terpenoids

[72]

27

Swertia chirata Roxb ex./Gentianaceae

Chirata

D. I. Khan

Seeds

Glycosides:

Amarogentin, amaroswerin, sweroside

[69]

28

Swertia paniculataWall./Gentianaceae

Momera

Allaivalley

Whole plant

NA

[80]

29

Tagete sminuta L./Compositae(= Asteraceae)

Marigold

Northern areas, Abbotabad

Seeds

Terpenoids, saponins, tannins, flavonoids, alkaloids

[45, 84]

30

Viburnum nervosumD. Don/Caprifoliaceae (= Adoxaceae)

NA

Azad Jammu Kashmir

Whole plant

Butilinol, oleanolic acid, butilinic acid, urosolic acid,α-amyrin, β-sitosterol

[85]

31

Vincetoxicum stocksii Ali &Khatoon/Apocynaceae (= Asclepiadaceae)

NA

Quetta

Whole plant

NA

[48]

32

Viola odorata L./Violaceae

Banafsha

Maradori valley

Whole plant

NA

[73]

33

Xanthium strumarium L./Compositae (= Asteraceae)

Desi Arinad

Allai valley

Leaves

NA

[80]

NA indicates data not available

Table 3

In-vitro screening of traditionally used anti-leishmanial and anti-malarial plants against Leishmania and Plasmodia parasites

Plant name

Part used

Parasite type

Extract

Concentration (µg/ml)

Inhibition (%)

Citation

Aloe vera

Leaves

Leishmania tropica

Crude methanol

25

15

[25]

50

27

75

43

100

66

Artemisia annua

Leaves

Plasmodium falciparum

Aqueous

0.095

50

[86]

Azadiracha indica

Leaves

Plasmodium falciparum

Ethanol

2.4

50

[36]

2.5

50

Asparagus asiaticus

Roots

Leishmania major

Crude methanol

25

22

[63]

50

34

250

42

500

51

Amphotericin B

(Reference drug as a control)

25

50

50

75

250

88

500

100

Citrullus colocynthis

Fruits

Leishmania major

Crude methanol

25

67

[35]

50

71

250

88

500

100

Juniperus excels

Fresh berries

Leishmania major

Crude methanol

25

49

[37]

50

58

250

88

500

97

Melia azedarach

Fruit

Leishmania tropica

Aqueous

1500

55.9

[86]

2500

67.4

5000

80.4

Moringa oleifera

Leaves

Plasmodium falciparum

Acetone

400

59.8

[31]

Nepeta praetervisa

Leaves

Leishmania major

Methanol

25

39

[62]

50

54

250

68

500

78

Peganum harmala

Seeds

Plasmodium falciparum

Ethanol

12.5

91

[58]

25

97.4

50

98.5

100

99.8

Chloroquine

NA

99.6

Perotis hordeiformis

Leaves

Leishmania major

Methanol

25

47

[65]

50

58

250

70

500

80

Amphotericin B

(Reference drug as a control)

25

50

50

75

250

88

500

100

Rhazya stricta

Leaves

Leishmania major

Crude methanol

25

65

[34]

50

70

250

92

500

100

Salvia bucharica

Leaves

Leishmania major

Crude methanol

25

44

[67]

50

40

250

59

500

75

Tamarix aphylla

Barks

Leishmania tropica

Crude methanol

25

20

[25]

50

28

75

54

100

84

Thuspeinanta brahuica

Leaves

Leishmania major

Crude methanol

25

40

[87]

50

58

75

70

100

82

Amphotericin B (Reference drug as a control)

25

50

50

75

75

88

100

100

Fig. 1

Artemisinin [47]

Fig. 2

a Physalin B [88], b physalin D [88], c physalin G [88], d physalin F [88]

Fig. 3

Sitosterol [89]

Medicinal plants used to treat leishmaniasis in Pakistan

Leishmaniasis is a neglected tropical disease. Visceral leishmaniasis and cutaneous leishmaniasis are the two main clinical types of leishmaniasis widespread in Pakistan. The incidence of visceral leishmaniasis has been reported from Dera Ismail Khan, Quetta, Tank, Hazara Division, Northern areas and Azad Jammu Kashmir [20]. About 90 % cases of cutaneous leishmaniasis have been reported from all the provinces of Pakistan [21]. The reported endemic areas of cutaneous leishmaniasis are Dir, Chitral, Swat, Mansehra, Dera Ghazi khan, Gilgat, Skardu, Abbotabad, Azad Kashmir, Chilas, Rawalpindi, Khuzdar, Jacobabad, Lasbela, Derabughti, Rajanpur, Quetta, Qila Saifullah, Qila Abdullah, Pishan, Dera Ismail Khan, Larkana and Dadu [22]. These areas are foot hills of mountainous range and situated in North, South and South-Western Pakistan covering about all the provinces including Azad Kashmir. Growth and development of vector sandfly is promoted by the environmental conditions of these endemic areas [23].

Most of the above mentioned regions of Pakistan are rural in nature lacking modern health and education facilities and inhabitants of these regions have low economic status due to least income sources. Moreover, rural people rely on their rich traditional knowledge for their primary health care due to high cost of allopathic drugs [19]. Present review showed that traditional people use 23 medicinal plants belonging to 19 families for the treatment of leishmaniasis (Table 1). Other areas of Pakistan are also known for containing variety of medicinal plants and classic traditional healing practices but scientific documentation has not been yet done. The most widely used plant families for the treatment of leishmaniasis in Pakistan are Lamiaceae (four plants), Liliaceae (two plants) and Asclepiadaceae (two plants). The family Lamiaceae and Liliaceae usually ranks high in ethnomedicinal studies not only in Pakistan but throughout the world [2426]. Perez et al. [27] also reported high number of plants belonging to Lamiaceae family having anti-parasitic activity including leishmaniasis. Present findings indicate that Lamiaceae family contains variety of anti-parasitic secondary metabolites and should be given focus in future studies. Other reasons of its wide use might be due to higher abundance of these plants in different regions and strong traditional beliefs [2830]. Almost all plant parts are found to have anti-leishmanial activity but most preferred parts in Pakistan are leaves, fruits, roots and aerial parts. Leaves are also the most focused part of plant in in vitro screening against leishmaniasis not only in Pakistan but other countries of world [31, 32]. Most of the metabolic processes take place in leaves result in production of different secondary metabolites; therefore, it might be attributed with its wider utilization for in vitro screening and traditional medicines [33]. In some areas like Quetta, Islamabad and Malakand whole plant is used to treat leishmaniasis which is major threat to the conservation status of these medicinal plants. People should be educated regarding proper harvesting of these valuable anti-leishmanial plants for sustainable utilization.

In-vitro activities of anti-leishmanial plants

Majority of the modern allopathic drugs of the world have developed on the basis of traditional knowledge of the people regarding medicinal plants. Among 23 medicinal plants used against leishmaniasis in Pakistan only 11 plants have been studied worldwide for their in vitro activity against L. major and L. tropica parasites and documented in the present review. Anti-leishmanial activity of medicinal plants has shown excellent activity against Leishmania parasite (Table 3). Different plant parts have been used for extract formation experimentally among which leaves, fruits and roots are most widely used parts. This result gives an indication about the reliability of traditional ethnomedicinal knowledge and efficacy of these practices. Different plant extracts like crude methanol and methanol have been used at different concentrations (µg/ml) for their efficacy against L. major and L. tropica but crude methanol extract is most commonly used [34]. Crude methanol extraction of plant parts is also practiced in other parts of the world. Crude methanolic extract of different plants have shown strong inhibition zone ranges from 65 to 100 % at different concentrations ranging from 25 to 500 µg/ml against L. major parasite [34, 35], while methanolic extract of different plant parts having concentration of about 25–500 µg/ml shown optimum inhibition zone ranging from 39 to 80 %. Aqueous extract of a plant have shown inhibition zone ranging from 55.9 to 80.4 % at concentration of about 25–5000 µg/ml [35, 36]. Plant extracts have been proved more effective against leishmaniasis as compared to allopathic drugs due to less toxicity [36]. Therefore it is imperative to investigate and explore medicinal plants scientifically for the development of novel anti-leishmanial drugs of strong efficacy. Experimental investigation of different plants have shown presence of phytochemical constituents such as alkaloids, flavonoids, carbohydrates, diterpenes, saponins, phenols and tannins that might be responsible for their inhibitory activities against Leishmania parasites [36, 37]. Very few studies conducted on the purification of pure compounds from above mentioned plants and should be given focus in future.

Medicinal plants used to treat malaria in Pakistan

In modern medical terms, malaria can be defined as infection caused by red blood cells parasite belonging to genus Plasmodium. Malaria is a major serious public health problem caused by P. falciparum and P. vivax, the two most prevalent Plasmodium species throughout the world. Approximately 64 % cases of malaria caused by P. vivax and about 36 % cases by P. falciparum in Pakistan [38]. According to WHO report about 1.6 million cases of malaria were reported in endemic areas per year [39]. The cases of malaria infection are reported from Sindh, Punjab, Khyber Pakhtunkhwa, Baluchistan and FATA areas. In these regions malaria often occurs in poor people because majority of population in these regions are rural with very low socioeconomic status. The environment of these areas promotes optimum growth of female Anopheles vector [40]. Reason behind high prevalence of malaria in poor people of Pakistan might be due to that malaria strike in the season when economic conditions are more difficult for the people. In Brazil 99 % malarial cases are reported and transmitted in Amazon region, where population consists of tribal people and immigrants from other regions [40]. History has proven traditional medicines to be the best source of effective anti-malarial e.g. Cinchona spp. and Artemisia annua L. [41].

Chloroquine is the most commonly used antibiotic for the treatment of malaria not only in Pakistan but throughout the world [38]. Low income status of poor people and emergence of antibiotic resistance of parasite encourage the use of traditional medicines for the treatment of malaria. Ethnomedicinal used of plants are common throughout the world including Pakistan [42]. Present review documented 33 anti-malarial plants traditional being use in Pakistan (Table 2). The most widely used plant families for the treatment of malaria in Pakistan are Asteraceae (9 plants), Gentianaceae, Lamiaceae and Asclepiadaceae (3 plants each). The medicinal plants belonging to these families are extremely used for medicinal purposes including anti-malarial purposes not only in Pakistan but throughout the world [4345] that might be due to greater availability or high traditional values of these plants in different regions. Traditional healers mostly used leaves for the preparation of ethnomedicinal recipes against malaria and these findings are not surprising because leaves are the most focused plant part throughout the world [44, 46]. In different regions of Pakistan mostly whole plant is used for the treatment of malaria due to the presence of important compounds. It is considered to be one of the major causes of extinction of highly valuable medicinal plants in many areas of Pakistan.

In-vitro activities of anti-malarial plants

In present review, among 33 medicinal plants used to treat malaria in Pakistan, only 4 plants have been investigated experimentally throughout the world for their in vitro activity against P. falciparum (Table 3). Only two plant parts, seeds and leaves have been used for extract preparation. Different plant extracts like acetone, aqueous and ethanol have been used scientifically at different concentrations (µg/ml) for their efficacy against P. falciparum [32, 37, 47]. Ethanol extraction of plants is also followed throughout the world due to its polar nature [42]. Ethanolic extracts of two different plants like Azadiracha indica and Peganum harmala have shown strong inhibition zone ranging from 50 to 99.8 % at concentration of about 2.4–100 µg/ml against P. falciparum. Acetone extract of Artemisia annua show 50 % inhibition zone at concentration of about 0.095 µg/ml and aqueous extracts of Moringa oleifera show optimum inhibition zone 59.8 % at concentration of 400 µg/ml. These results show the strong efficacy of plants extracts against P. falciparum in comparison with standard drug. Phytochemical screening of different plant extracts have not been studied in detail but experimentally studied plant parts mostly contain alkaloids, flavonoids, saponins, tannins and terpenoids that might be responsible for anti-parasitic activities of these plants. Other plants needs in vitro exploration and phytochemical screening that could lead toward extraction of some novel compounds/drugs against Plasmodium parasite.

Medicinal plants with both anti-leishmanial and anti-malarial potential

Three plants Melia azedarach L. (Meliaceae), Vincetoxicum stocksii L. (Asclepiadaceae) and Carum copticum L. (Umbelliferae) have been used for the treatment of both leishmaniasis and malaria which show their high potential for anti-parasitic activity [19, 48]. But in vitro activity of only one plant M. azedarach has been investigated experimentally against Leishmania parasite. Aqueous extract of fruit of M. azedarach showed strong inhibition zone of about 55.9, 67.4 and 80.4 % against L. tropica at three different concentrations 1500, 2500 and 5000 µg/ml, respectively (Table 3). Present finding is very interesting because it gives an indication about strong efficacy of these candidate medicinal plants for future research against malaria.

Active phyto-compounds against Leishmania and Plasmodia parasites

Only three compounds isolated from traditionally used anti-leishmanial and anti-malarial plants of Pakistan were investigated for their anti-parasitic activity.

Artemisinin

In present review four plant species of Artemisia used in Pakistan to treat malaria (Table 2). The genus Artemisia has great importance in pharmaceutics as it is used in traditional medicines to treat various diseases especially malaria not only in Pakistan but throughout the world [4951]. In-vitro study of Artemisia plants shows that they contain an important chemical compound Artemisinin, Sesquiterpenoid lactone. Artemisinin (Fig. 1) is extracted from the leaves of Artemisia and known to have best antimalarial activity. WHO recommended the Artemisinin combination therapy for the treatment of malaria caused by P. falciparum [2, 52]. Artemisia annua, a good source of Artemisinin is endemic plant of China and used as folk medicine to treat malaria for about 2000 years [52]. Artemisinin have also been reported for good anti-viral, anti-cancer and anti-leishmanial activity [53].

Physalins

Several physalins (Steroidal lactone) were isolated from various species of genus Physalis belonging to the family Solanaceae. Physalins (Fig. 2) have both anti-leishmanial and anti-malarial potential [54, 55]. Four types of physalins B, D, G and F (Fig. 2a–d) were isolated from Physalis angulata. The in vitro and in vivo activity of physalins B and F showed potent anti-leishmanial activity against various Leishmania parasites like L. brazillenesis, L. amazonensis, L. major and L. chagasi [55, 56]. Physalins B, D, G and F have also been reported for their anti-malarial activity against P. falciparum [54].

Sitosterol

Aloe barbadense is an important medicinal plant having bioactive compounds reported for their anti-leishmanial activity. Sitosterol (Fig. 3) is an important compound extracted from the leaves of Aloe vera. It inhibits the growth of promastigotes of L. donovani, a causative agent for life threatening visceral leishmaniasis disease. The active components of Aloe vera target the CDC42 protein in comparison with a natural inhibitor Sacramine B [57].

Antibiotic resistance of Leishmania and Plasmodia parasites

Literature review showed that both Leishmania and Plasmodium parasites have shown resistance to various antibiotics that are being used for the treatment of leishmaniasis and malaria. Various antibiotics have been used for the treatment of malaria worldwide like chloroquine, halofantrine, pyrimethamine, mefloquine, quinine and artemisinin [2, 4]. The in vitro investigation of chloroquine showed resistance range from 69.8 to 99.6 % against P. falciparum [58, 59].

The most commonly used drugs for the treatment of leishmaniasis are pentavalent antimonials like sodium stibogluconate and meglumine antimoniate. Beside these many other drugs like amphotericin, ambisome (lipid formulation of amphotericin), miltefosine (impavido), pentamidine and paromomycin were discovered to treat leishmaniasis [60]. All types of Leishmania parasites show resistance against one drug or other [19, 60]. Due to emerging potential of drug resistance of parasites, high cost of allopathic drugs and their side effects encourages the use of traditional medicines among local population worldwide.

Conclusions and future recommendations

Pakistan has tremendous potential regarding the use of ethnomedicines for the treatment of multiple diseases including malaria and leishmaniasis. This review provides a scientific rationale for the traditional uses of medicinal plants against these diseases. Traditional healers of different regions have strong knowledge to utilize medicinal plants. In-vitro screening of traditionally used anti-parasitic plants have proven the efficacy of such plants. Crude plant extracts, methanolic and other extracts were effective in antimalarial and anti-leishmanial activities. Mostly, leaves of documented plants are traditionally used and also for in vitro screening. Different classes of compounds exist in the documented plants including alkaloids, flavonoids and terpenoids. Very few compounds have so far been isolated from the documented plants and tested in vitro against studied leishmanial and malarial parasites. In pure compounds, ursolic acid and cucurbitacin in C. colocynthis while glycosides and alkaloids isolated from R. stricta possess anti-leishmanial activities. On the other side, compounds such as limonoids (gedunin) from A. indica while β-carboline alkaloids, Harmaline and Harmine isolated from P. harmala have proven in vitro and in vivo anti-plasmodial activities. Hence, these plant species must be explored for the identification of more such compounds to be used against Leishmania and Plasmodium parasites. Moreover in the present era, parasites are showing resistance to common allopathic drugs, while on the other side medicinal plants have proven their effectiveness as anti-parasitic drugs. It is therefore imperative to conduct future studies on the unexplored documented plants both in vitro and in vivo for the development of novel drugs. On the basis of findings in this review, following recommendations are suggested:
  • Ethnomedicinal studies provide baseline information for future scientific research, therefore it is recommended to expedite exploration of anti-parasitic plants not only in Pakistan but throughout the world.

  • Traditional healers mostly use Lamiaceae and Asteraceae families for the treatment of leishmaniasis and malaria, respectively. It invites the attention of worldwide researchers to explore species belonging to these families both phytochemically and pharmacologically.

  • Among all plant parts, leaves have taken more focus both in traditional medicines as well as in in vitro studies. Other plant parts should also be brought under the spotlight for the potential discovery of different compounds.

  • Different extracts of documented plants have been used worldwide against leishmaniasis and Plasmodia. Crude extracts have been given more preference against leishmaniasis while only very few plants have been tested in vitro against Plasmodia. Extracts like methanolic, ethanolic, n-hexane should also be valued against both parasites that could be helpful in extraction of some novel compounds.

  • More attention should be given towards the isolation of pure compounds from these plants, and their in vitro investigations against leishmanial and malarial parasites.

  • In-vivo studies should also be brought under the focus in order to pharmacologically validate these traditional plants.

  • Action mechanism of different extracts and pure compounds on the studied parasites should also be studied in future research.

  • Toxicity of these plants should also be tested on living system that would be helpful in proving the reliability of traditional medicines.

Abbreviations

WHO: 

World health organization

FATA: 

Federally Administered Tribal Areas

Declarations

Authors' contributions

All authors contributed to this work. All authors have read and approved the final manuscript.

Acknowledgements

The authors are indebted to all those who worked and are currently working on various aspects related to the prevention of Malaria and Leishmaniasis.

Competing interests

The authors declare that they have no competing interests.

Availability of data and materials

We declare that the data supporting the conclusions of this article are fully described within the article.

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

Authors’ Affiliations

(1)
Key Laboratory of Mountain Ecological Restoration and Bioresource Utilization & Ecological Restoration Biodiversity Conservation Key Laboratory of Sichuan Province, Chengdu Institute of Biology, Chinese Academy of Sciences
(2)
Department of Botany, Kohat University of Science and Technology
(3)
Department of Zoology, Kohat University of Science and Technology
(4)
Department of Biotechnology, Quaid-i-Azam University Islamabad

References

  1. Ngurea PK, Kimutaib A, Zipporah W, Rukungad G, Tonui WK. A review of Leishmaniasis in Eastern Africa. J Nanjing Med U. 2009;23:79–86.View ArticleGoogle Scholar
  2. Shahinas D, Folefoc A, Pillai DR. Targeting Plasmodium falciparum Hsp90: towards reversing antimalarial resistance. Pathogens. 2013;2:33–54.View ArticlePubMedPubMed CentralGoogle Scholar
  3. Pradeep D, Steven S, Philippe D, Atul M, Roshan T, Niyamat AS, Dipika S, Arvind P, Rhonda S. Annual incidence of visceral leishmaniasis in an endemic area of Bihar, India. Trop Med Int Health. 2010;10:1365.Google Scholar
  4. Nahrevanian H, Milan BS, Kazemi M, Hajhosseini R, Mashhadi SS, Nahrevanian S. Antimalarial Effects of Iranian Flora Artemisia sieberi on Plasmodium berghei In vivo in mice and phytochemistry analysis of its herbal extracts. Malar Res Treat. 2012. doi:https://doi.org/10.1155/2012/727032.Google Scholar
  5. Uzun S, Uslular C, Yucel A, Acar MA, Ozpoyraz M, Memisoglu HR. Cutaneous leishmaniasis evaluation of 3074 cases in Cukurova region of Turkey. Br J Dermatol. 1999;140:347–50.View ArticlePubMedGoogle Scholar
  6. Alavi-Naini R, Fazaeli A, Dempsey OT. Topical treatment modalities for old world cutaneous leishmaniasis: a review. Prague Med Rep. 2012;113:105–18.View ArticlePubMedGoogle Scholar
  7. Zand M, Narasu ML. Vaccination against leishmaniasis. Ann Biol Res. 2013;4:170–4.Google Scholar
  8. Croft SL, Coombs GH. Leishmaniasis-current chemotherapy and recent advances in the search for novel drugs. Trends Parasitol. 2003;19:502–8.View ArticlePubMedGoogle Scholar
  9. Hardman JG, Limbird LE. Drugs used in the chemotherapy of malaria in The Goodman and Gilman’s Pharmacologicalbases of Therapeutics. 10th ed. New York: McGraw-Hill; 2001.Google Scholar
  10. Leslie T, Mikhail A, Mayan I, Cundill B, Anwar M, Bakhtash SH, Mohammed N, Rahman H, Zekria R, Christopher JMW. Mark Rowland Rapid diagnostic tests to improve treatment of malaria and other febrile illnesses: patient randomised effectiveness trial in primary care clinics in Afghanistan. Br Med J. 2014;348:1–13.View ArticleGoogle Scholar
  11. Fidock DA, Rosenthal PJ, Croft SL, Brun R, Nwaka S. Antimalarial drug discovery: efficacy models for compound screening. Nat Rev Drug Discov. 2004;3:509–20.View ArticlePubMedGoogle Scholar
  12. Douradinha B, Doolan DL. Harnessing immune responses against Plasmodium for rational vaccine design. Trends Parasitol. 2011;27:274–83.View ArticlePubMedGoogle Scholar
  13. Bhutto AM, Soomro RA, Nonaka S, Hashiguchi Y. Detection of new endemic areas of cutaneous leishmaniasis in Pakistan: a 6-year study. Int J Dermatol. 2003;42:543–8.View ArticlePubMedGoogle Scholar
  14. Ikram AU, Zahra NB, Shinwari ZK, Qaiser M. Ethnomedicinal review of folklore medicinal plants belonging to family Apiaceae of Pakistan. Pak J Bot. 2015;47:1007–14.Google Scholar
  15. Tiuman TS, Santos AO, Nakamura TU, Filho BPD, Nakamura CV. Recent advances in Leishmaniasis treatment. Int J of Infect Dis. 2011;15:525–32.View ArticleGoogle Scholar
  16. Qasim M, Abideen Z, Adnan MY, Ansari R, Gul B, Khan MA. Traditional ethno-botanical uses of medicinal plants from coastal areas of Pakistan. J Coastal Life Med. 2014;2:22–30.Google Scholar
  17. Jeruto P, Lukhoba C, Ouma G, Otieno D, Mutai C. An ethnobotanical study of medicinal plants used by the Nandi people in Kenya. J Ethnopharmacol. 2008;116:370–6.View ArticlePubMedGoogle Scholar
  18. Shah SMH, Shah SMM, Nisar M, Khan FA, Ali M. KhanI. Antimicrobial activities of medicinal plants used in folk remedies in Pakistan. J Pharm Res. 2012;5:2057–60.Google Scholar
  19. Khan I, Yasinzai MM, Mehmood Z, Ilahi I, Khan J, Khalil AT, Saqib MS, Rahman WU. Comparative study of green fruit extract of Melia azedarach Linn. with its ripe fruit extract for antileishmanial, larvicidal, antioxidant and cytotoxic activity. Am J Phytomed. Clin Ther. 2014;2:442–54.Google Scholar
  20. Kakarsulemankhel JK. Leishmaniasis in Pak-Afghan Region. Int J Agric Biol. 2011;13:611–20.Google Scholar
  21. Ali N, Afrin F. Protection of mice against Visceral Leishmaniasis by immunization with promastigotes antigen incorporated in liposomes. J Parasitol. 1997;83:70–5.View ArticlePubMedGoogle Scholar
  22. Shah NA, Khan MR, Nadhman A. Antileishmanial, toxicity, and phytochemical evaluation of medicinal plants collected from Pakistan. BioMed Res Int. 2014. doi:https://doi.org/10.1155/2014/384204.Google Scholar
  23. Durrani AZ, Durrani HZ, Kamal N. Prevalence of Leishmania in sandfly in Pakistan. Pak J Zool. 2012;44:61–5.Google Scholar
  24. Nasir E, Ali SI. Flora of Pakistan, vol. 175. Karachi: Fakhri Printing Press; 1986. p. 200.Google Scholar
  25. Iqbal H, Khattak B, Ayaz S, Rehman A, Ishfaq M. Comparative efficacy of Aloe vera and Tamarix aphylla against cutaneous leishmaniasis. Int J Basic Med Sci Pharm. 2012;2:42–5.Google Scholar
  26. Serakta M, Djerrou Z, Djaalab HM, Riachi FK, Hamimed S, Trifa W, Belkhiri A, Edikra N, Pacha YH. Antileishmanial activity of some plants growing in Algeria Juglans Regia, Lawsonia Inermis and Salvia Officinalis. Afr J Trad Complem Altern Med. 2013;10:427–30.Google Scholar
  27. Perez SG, Ramos-lopez MA, Sanchez-miranda E, Fresan-Orozoco MC, Perez-Ramos J. Antiprotozoa activity of some essential oils. J Med Plants Res. 2012;6:2901–8.Google Scholar
  28. Kirtikar KR, Basu BD. Indian medicinal plants. Allahad: Indian Press; 1981. p. 1031.Google Scholar
  29. Weiner M, Weiner JA. Herbs that heal. Mill Valley. 1994;75:270–2.Google Scholar
  30. Ali MI, Shalaby NM, Elgamal MH, Mousa AS. Antifungal effects of different plant extracts and their major components of selected Aloe species. Phytother Res. 1999;13:401–7.View ArticlePubMedGoogle Scholar
  31. Patel JP, Gami B, Patel K. Evaluation of in vitro Schizonticidal properties of acetone extract of Some Indian medicinal plants. Adv Biol Res. 2010;4:253–8.Google Scholar
  32. Adebayoa JO, Krettli AU. Potential antimalarials from Nigerian plants: a review. J Ethnopharmacol. 2011;133:289–302.View ArticleGoogle Scholar
  33. Murad W, Azizullah A, Adnan M, Tariq A, Khan KU, Waheed S, Ahmed A. Ethnobotanical assessment of plant resources of Banda Daud Shah, District Karak. Pakistan. J Ethnobiol Ethnomed. 2013;9:1–10.View ArticleGoogle Scholar
  34. Khan MJ, Baloch NU, Nabi S, Ahmed N, Bazai Z, Yasinzai M, Yasser MSA, Kahraman A. Antileishmanial, cytotoxic, antioxidant activities and phytochemical analysis of Rhazya stricta Decne leaves extracts and its fractions. Asian J Plant Sci Res. 2012;2:593–8.Google Scholar
  35. Baloch N, Nabi S, Kakar AM, Wajid Z, Alkharaman YMSA. In-vitro antileishmanial, antitumor, cytotoxic activities and phytochemical analysis of Citrullus colocynthis fruit extract. Int J Med Aroma Plants. 2013;3:78–84.Google Scholar
  36. Abdullah IH, Farooq A, Shazia R, Poonam SN, Omar J, Satyajit DS. Composition, antioxidant and chemotherapeutic properties of the essentialoils from two Origanum species growing in Pakistan. Braz J Pharmacog. 2011;21:943–52.Google Scholar
  37. Nabi S, Ahmed S, Khan MJ, Bazai Z, Yasinzai M, Yasser MSA, Kahraman A. In vitro antileishmanial, antitumor activities and phytochemical studies of methanolic extract and its fractions of Juniperus Excelsa Berries. World Appl Sci J. 2012;19:1495–500.Google Scholar
  38. Khattak AA, Venkatesan M, Khatoon L, Ouattara A, Kenefic LJ, Nadeem MF, Nighat F, Malik SA, Christopher VP. Prevalence and distribution of human Plasmodium infection in Pakistan. Malaria J. 2013;12:297.View ArticleGoogle Scholar
  39. WHO. World malaria report 2012. Geneva: World Health Organization; 2013.Google Scholar
  40. Willcox ML, Bodeker G. Traditional herbal medicines for Malaria. Br Med J. 2004;329:1156–9.View ArticleGoogle Scholar
  41. Willcox ML, Bodeker G, Rasoanaivo P. Traditional medicinal plants and Malaria. Boca Raton: CRC Press; 2004.Google Scholar
  42. Mojarrab M, Shiravand A, Delazar A, Afshar, F.H. Evaluation of in vitro antimalarial activity of different extracts of Artemisia aucheri Boiss and A. armeniaca Lam and fractions of the most potent extracts. Sci World J. 2014. doi: https://doi.org/10.1155/2014/825370.
  43. Azas N, Laurencin N, Delmas F, Di GC, Gasquet M, Laget M, Timon-David P. Synergistic in vitro antimalarial activity of plant extracts used as traditional herbal remedies in Mali. Parasitol Res. 2002;88:165–71.View ArticlePubMedGoogle Scholar
  44. Titanji VPK, Zofou DS, Ngemenya MN. The antimalarial potential of medicinal plants used for the treatment of malaria in Cameroonian Folk Medicine. Afr J Trad Complem Altern Med. 2008;5:302–21.Google Scholar
  45. Sadia S, Khalid S, Qureshi R, Tagetes Bajwa AA, Minuta L. A useful underutilized plant of family Asteraceae: a Review. Pak J Weed Sci Res. 2013;19:179–89.Google Scholar
  46. Luize PS, Tiuman TS, Morello LG, Maza PK, Nakamura TU, Filho PBD, Cortez DAG, Mello JCP, Nakamura CV. Effects of medicinal plant extracts on growth of Leishmania (L.) amazonensis sand Trypanosoma cruzi. Braz J Pharma Sci. 2005;41:85–94.Google Scholar
  47. Ploypradith P. Development of artemisinin and its structurally simplified trioxane derivatives as antimalarial drugs. Acta Trop. 2004;89:329–42.View ArticlePubMedGoogle Scholar
  48. Mansoor A, Ibrahim MA, Zaidi MA, Ahmed M. Antiprotozoal activities of Vincetoxicum stocksii and Carum copticum. Bangladesh J Pharmacol. 2011;6:51–4.View ArticleGoogle Scholar
  49. Arab HA, Rahbari S, Rassouli A, Moslemi MH, Khosravirad F. Determination of artemisinin in Artemisia sieberiand anticoccidial effects of the plant extract in broiler chickens. Trop Anim Health Prod. 2006;38:497–503.View ArticlePubMedGoogle Scholar
  50. Romero MR, Serrano MA, Vallejo M, Efferth T, Alvarez M, Marin JJ. Antiviral effect of artemisinin from Artemisia annua against a model member of the Flaviviridae family, the bovine viral diarrhea virus (BVDV). Planta Med. 2006;72:1169–74.View ArticlePubMedGoogle Scholar
  51. Willoughby JA, Sundar SN, Cheung M, Tin MA, Modiano J, Firestone GL. Artemisinin blocks prostate cancer growth and cell cycle progression by disrupting Sp1 interactions with the cyclin-dependent kinase-4 (CDK4) promoter and inhibiting CDK4 gene expression. J Biol Chem. 2009;284:2203–13.View ArticlePubMedPubMed CentralGoogle Scholar
  52. Mannan A, Ahmed I, Arshad W, Asim MF, Qureshi RA, Hussain I, Mirza B. Survey of artemisinin production by diverse Artemisia species in northern Pakistan. Malar J. 2010;9:310.View ArticlePubMedPubMed CentralGoogle Scholar
  53. Sen R, Bandyopadhyay S, Dutta A, Mandal G, Ganguly S, Saha P, Chatterjee M. Artemisinin triggers induction of cell-cycle arrest and apoptosis in Leishmania donovani promastigotes. J Med Microbiol. 2007;56:1213–8.View ArticlePubMedGoogle Scholar
  54. Guimarães ET, Lima MS, Santos LA, Ribeiro IM, Tomassini TBC, Santos RR, Milena BPS. Effects of seco-steroids purified from Physalis angulata L., Solanaceae, on the viability of Leishmania sp. Braz J Pharmacog. 2010;20:945–9.Google Scholar
  55. Elisalva T. Guimara, Milena S, Lima, Luana A, Santos, Ivone M, Ribeiro, Therezinha BC, Tomassini, Santos RR, Washington LC, Santos, Milena BPS. Activity of physalins purified from Physalis angulata in in vitro and in vivo models of cutaneous leishmaniasis. J Antimicrob Chemother. 2009;64:84–7.View ArticleGoogle Scholar
  56. Matheus SS, Menezes MND, Krettli AU, Ribeiro IM, Tomassini TC, Santos RR, Azevedo WF, Soares MB. Antimalarial activity of physalins B, D, F, and G. J Nat Prod. 2011;74:2269–72.View ArticleGoogle Scholar
  57. Priyanka BM, Vennila J, Ganesh S. An in-silicoapproach to identify drug targetable protein in visceral leishmaniasis using GC MS extracted active components from Aloe vera. Adv Appl Sci Res. 2011;2:426–31.Google Scholar
  58. Nateghpour M, Sharbatkhori M, Edrissian GH, Souri E, Mohebali M, Akbarzadeh K, Haghi AM, Satvat M, Rahimi A. Assessment of in vitro activity of Peganum harmala extract on Plasmodium falciparum growth compared with chloroquine. Pak J Biol Sci. 2006;9:214–6.View ArticleGoogle Scholar
  59. Olasehinde GI, Ojurongbe O, Adeyeba AO. In-vitro studies on the sensitivity pattern of Plasmodium falciparum to anti-malarial drugs and local herbal extract. Malar J. 2014;13:63.View ArticlePubMedPubMed CentralGoogle Scholar
  60. Monzote L. Current treatment of leishmaniasis. Open Antimicrob Agents J. 2009;1:9–19.Google Scholar
  61. Marwat SK, Rehman FU, Khan MA, Ahmad M, Zafar M, Ghulam S. Medicinal folk recipes used as traditional phytotherapies in District Dera Ismail Khan. KPK Pakistan. Pak J Bot. 2011;43:1453–62.Google Scholar
  62. Baloch N, Nabi S, Yasser MSA, Kahraman A. In-vitro antileishmanial, cytotoxic, antioxidant activities and phytochemical analysis of Berberis baluchistanica roots extracts and its fractions. J Phytopharmacog. 2013;4:282–7.Google Scholar
  63. Baloch N, Nabi S, Bashir S, Yasser MSA, Kahraman A. In-vitro antileishmanial, cytotoxic activity and phytochemical analysis of Nepeta praetervisa leaves extract and its fractions. Int J Pharm Pharma Sci. 2013;5:475–8.Google Scholar
  64. Ahmad B, Ali N, Bashir S, Choudhary MI, Azam S, Khan I. Parasiticidal, antifungal and antibacterial activities of Onosma griffithii Vatke. Afr J Biotechnol. 2009;8:5084–7.Google Scholar
  65. Baloch N, Nabi S, Yasser MSA, Kahraman A. In-vitro antileishmanial, cytotoxic, antioxidant activities and their phytochemical analysis on methanolic extract and it is fractions of Perotis hordeiformis leaves. Int J Pharma Sci Rev Res. 2013;22:191–5.Google Scholar
  66. Rahman AU. Samreen, Wahab A, Choudhary MI. Discovery of leishmanicidal agents from medicinal plants. Pure Appl Chem. 2008;80:1783–90.Google Scholar
  67. Khan A, Khan MJ. In vitro antileishmanial, cytotoxic and antioxidant activities of Salvia bucharica leaves extract and its fractions. Int J Basic Appl Sci. 2013;13:74–8.Google Scholar
  68. Choudhary MI, Adhikri A, Rehman AU. Antileishmanial steroidal alkaloids from roots of Sarcococca coriacea. J Chem Soc Pak. 2010;32:799–802.Google Scholar
  69. Sher A. Antimicrobial activity of natural products from medicinal plants. Gomal J Med Sci. 2009;7:1–4.Google Scholar
  70. Kakar AM, Khan AA, Nabi S, Kakar MA, Yasinzai M, Ymsa AK. In vitro antileishmanial, cytotoxic activity and phytochemical analysis of Thuspeinanta Brahuica leaves extract and its fractions. Int J Biol Pharm Appl Sci. 2013;2:520–8.Google Scholar
  71. Bashir A, Ali N, Bashir S, Choudhary MI. Biological activities of aerial parts of Tylophora hirsuta Wall. Afr J Biotechnol. 2009;8:4627–31.Google Scholar
  72. Wadood A, Ghufran M, Jamal SB, Naeem M, Khan A, Ghaffar R. Asnad. Phytochemical analysis of medicinal plants occurring in local area of Mardan. Biochem. Anal Biochem. 2013;2:1–4.Google Scholar
  73. Ishtiaq M, Ahmed F, Maqbool M, Hussain T. Ethnomedicinal inventory of flora of Maradori Valley, district Forward Khahuta, Azad Kashmir. Pakistan. Am J Res Commun. 2013;1:239–61.Google Scholar
  74. Khan RU, Mehmood S, Khan SU, Jaffar F. Ethnobotanical study of food value flora of district Bannu Khyber Pakhtunkhwa. Pakistan. J Med Plant Stud. 2013;1:93–106.Google Scholar
  75. Hayat MQ, Khan MA, Ashraf M, et al. Ethnobotany of the Genus Artemisia L. (Asteraceae) in Pakistan. Ethnobot Res Appl. 2009;7:147–62.View ArticleGoogle Scholar
  76. Mahmood A, Mahmood A, Hussain I, Jabeen S. Indigenous medicinal knowledge of medicinal plants of Barnala area district Bhimber. Pakistan. Int J Med Arom Plants. 2011;1:294–301.Google Scholar
  77. Azhar MF, Siddique MT, Ishaque M, Tanveer A. Study of ethnobotany and indigenous use of Calotropis Procera (Ait.) in Cholistan desert, Punjab. Pak J Agric Res. 2014;52:117–26.Google Scholar
  78. Hadi F, Razzaq A, Rahman AU, Rashid A. Ethnobotanical notes on woody plants of Rech Valley, Torkhow, district Chitral, Hindu-Kush range, Pakistan. Scholarly J Agric Sci. 2013;3:468–72.Google Scholar
  79. Zafar F, Jahan N, Rahman KU, Zafar WUI, Aslam S. Comparative evaluation of phytochemical, mineral and vitamin contents of Gemmo modified extracts and leaves of two indigenous medicinal plants. Int J Agric Biol. 2014;16:911–6.Google Scholar
  80. Haq F. The ethno botanical uses of medicinal plants of Allai Valley, Western Himalaya Pakistan. Int J Plant Res. 2012;2:21–34.View ArticleGoogle Scholar
  81. Fatima T, Sajid MS, Hassan MJU, Siddique RM, Iqbal Z. Phytomedicinal value of Moringa oleifera with special reference to antiparasitics. Pak J Agric Sci. 2014;51:251–62.Google Scholar
  82. Afzal S, Afzal N, Awan MR, Khan TS, Gilani A, Khanum R, Tariq S. Ethno-botanical studies from Northern Pakistan. J Ayub Med Coll Abbottabad. 2009;21:52–7.PubMedGoogle Scholar
  83. Khan H, Saeed M, Muhammad N, Tariq SA, Ghaffar R, Gul F. Antimalarial and free radical scavenging activities of aerial parts of Polygonatum Verticillatum (L.) All and identification of chemical constituents by Gc-Ms. Pak J Bot. 2013;45:497–500.Google Scholar
  84. Shahzadi I, Hassan A, Ummara W, Khan UW, Shah MM. Evaluating biological activities of the seed extracts from Tagetes minuta L. found in Northern Pakistan. J Med Plants Res. 2010;4:2108–12.Google Scholar
  85. Awan ZI, Rehman HU, Minhas FA, Awan AA. Antiplasmodial activity of compounds isolated from Viburnum nervosum. Int J Pharma Sci Invent. 2013;2:19–24.Google Scholar
  86. Gueye PEO, Diallo M, Deme AB. Tea Artemisia annua inhibited by Plasmodium falciparum isolates collected in Pikine. Senegal. Afr J Biochem Res. 2013;7:107–13.Google Scholar
  87. Kakar AM, Khan AA, Nabi S, Kakar MA, Yasinzai M, Al-Kahraman YMSA. In-vitro Antileishmanial, Cytotoxic Activity and Phytochemical Analysis of Thuspeinanta brahuica leaves extract and its fractions. IJPBAS. 2013;2:520–8.Google Scholar
  88. Soares MBP, Brustolim D, Santos LA, Bellintani MC, Paiva FP, Ribeiro YM, Tomassini TCB, Santos RRD. Physalins B, F and G, seco-steroids purified from Physalis angulata L., inhibit lymphocyte function and allogeneic transplant rejection. Int J Immunopharmacol. 2006;6:408–14.View ArticleGoogle Scholar
  89. Chaturvedula VSP, Prakash I. Isolation of Stigmasterol and β-Sitosterol from the dichloromethane extract of Rubus suavissimus. Int Curr Pharm J. 2012;1(9):239–42.View ArticleGoogle Scholar

Copyright

© The Author(s) 2016