Skip to main content
NCBI home page
Journal of Parasitic Diseases: Official Organ of the Indian Society for Parasitology logoLink to Journal of Parasitic Diseases: Official Organ of the Indian Society for Parasitology
. 2014 Feb 16;39(4):663–672. doi: 10.1007/s12639-014-0436-4

Herbal extract targets in Leishmania tropica

Bassim I Mohammad 1, Maani N Al Shammary 3, Roaa H Abdul Mageed 4, Nasser Ghaly Yousif 1,2,✉
PMCID: PMC4675586  PMID: 26688631

Abstract

The present study aims to investigate the effect of some herbal extract such as phenolic compounds on the viability of Leishmania tropica promastigotes in vitro. Four tested chemical agents (caffeic acid (CA), ferulic acid (FA), syringic acid (SA) and 4-hydroxybenzoic acid (4-HBA)) were used in this study. The viability of Leishmania tropica promastigotes was investigated under five different concentrations (10, 15, 20, 25 and 30 mg/ml) of each agent after (72 h). CA was the most active agent on the promastigotes viability after 72 h exposure to 30 mg/ml concentration so that the parasiticidal effect reach (53 × 104) promastigote/ml. FA is the second agent in parasiticidal effect that parasiticidal effect reach to (50 × 104 promastigote/ml) at a concentration (30 mg/ml), 4-HBA is the third agent in parasiticidal effect that reach to (48 × 104 promastigote/ml) at a concentration (30 mg/ml), SA is the weakest agent in parasiticidal activity that reach to (44 × 104 promastigote/ml) at a concentration (30 mg/ml). It can be concluded that (CA, FA, SA and 4-HBA) possess acidal effect on the Leishmania tropica promastigotes in vitro.

Keywords: Leishmania tropica promastigotes, Phenolic compounds, Parasiticidal effect, Sodium stibogluconate

Introduction

Leishmaniasis is a disease caused by obligate intracellular, kinetoplastid protozoa of the genus Leishmania (Trypanosomatidae) (Roberts 2006). Leishmania has two stages in its life cycle amastigote stage in mammalian host and promastigote stage in vector (Osman et al. 2000). Natural transmission may be zoonotic or anthroponotic, and it is usually by the bite of a phlebotomine sand fly species (Killick-Kendrick 1990; Ready 2008). The disease manifest itself in variety of clinical forms, ranging from self-healing cutaneous lesion to the more serious, potentially fatal visceralizing form, and include the metastasize mucocutaneous form and the post kala-azar dermal Leishmaniasis (Thakur 2000). Leishmaniasis is one of the leading causes of morbidity and mortality and it has been found to be a major global health problem (WHO 2007). It is endemic in 88 countries, with an estimated yearly incidence of 1–1.5 million cases of cutaneous leishmaniasis and 500,000 cases of visceral leishmaniasis, three hundred and fifty million people are estimated to be at risk, and there is an overall prevalence of 12 million cases (Desjeux 1996). Cutaneous and mucocutaneous leishmaniasis are more prevalent in Afghanistan, Iraq, Saudi Arabia and some Latin American countries (WHO 2007).

Proven therapies against human leishmaniasis include pentavalent antimonials (sodium stibogluconate and meglumine antimonite), amphotericin B, pentamidine, miltefosine and paromomycin (Berman 1996, 1997). These drugs are unsatisfactory because of their limited efficacy, frequent side effects and increasing drug resistance. Development of resistance by the parasites has been reported (Ephros et al. 1997; Lira et al. 1999; Boelaert et al. 2002). A search for a new active compound with potential Leishmanicidal property remains essential for the development of a new antileishmanial therapy. Extracts from medicinal plants are being widely tested for Leishmanicidal activity (Takahashi et al. 2004).

Phenolic compounds are a group of chemical compounds that are widely distributed in nature. They are a chemical family whose members have one or more hydroxyl groups attached directly to an aromatic ring. Fruits, vegetables, spices and aromatic herbs are natural sources of phenolic compounds. There is significant evidence that phenolic compounds have positive effect on the human health with many recently published studies reporting anti-inflammatory, cardio protective activities, anti-allergic, anti-carcinogenic and UV filtering properties, antioxidant as well as anti-microbial properties (Waterman and Mole 1994). Therefor in the present study aims to investigate the effect of some herbal extract such as phenolic compounds on the viability of Leishmania tropica promastigotes in vitro.

Materials and methods

Leishmania tropica isolate was obtained from a research unit in the College of Medicine, AL-Nahrain University, Baghdad, Iraq. It was isolated from infected humans. The strain was diagnosed by isoenzyme assay and cultivated on biphasic culture medium at 25 °C. It’s composed of two phases (solid phase and liquid phase). This media is used for cultivation and continuation of promastigotes stage of Leishmania (Kagan and Norman 1970; Dawson et al. 1969).

RPMI 1640 (Roswell park memorial institute medium)

This media was prepared by adding 10.4 g of powder media in 900 ml of distill water and 100 ml of preheated fetal calf serum (55 °C for 50 min) then added 1 ml of previously prepared antibiotic solution and the pH was justified to 7.2. Sterilization has been done by Nalgen filter of 0.22 micrometer then distributed 5 ml of media in sterile tube of 10 ml size and incubated in 37 °C for 24 h to avoid the contamination tubes and the sterile tubes put in 4 C till use (Ranga and Julia 1997).

Growth curve

Promastigotes were cultured in RPMI-1640 with 10 % fetal calf serum medium and incubated in 26 °C. Counting the promastigote each 24 h was performed to determine logarithmic growth phase. The promastigote increased in number in the first, second, third days and reach the peak in the fourth day, the fourth day represent the logarithmic phase (Fig. 1).

Fig. 1.

Fig. 1

Open in a new tab

Line graph showing the mean count of parasites in culture within 5 days after initial inoculation

Preparation of samples

Assays on L. tropica promastigote were performed as follows

Promastigote were cultured in RPMI-1640 medium with 10 % fetal calf serum and incubated in 26 °C, promastigotes in the logarithmic growth phase were then added to each well of a 96-microtiter plate (100 μL). Different concentrations of agent were added on the parasite in each well (50 μL). Further two controls samples were used included: growth medium without agent and growth medium containing studied the anti-Leishmanial drug (Sodium stibogluconate) (20 μg/ml) (Darrell et al. 2002). The plate was incubated in 26 °C for 72 h. Growth was measured in each well through counting the promastigotes after (72 h) by the conventional slide chamber method (Carrio et al. 2001). The parasiticidal effect, the rate of parasitic killing after adding a certain dose of a specific experimental drug, was calculated as the number of dead parasites/total number of parasites (dead + alife) multiplied by 100. This index ranges between a minimum of zero to a maximum of 100. The higher the index the stronger is the effect (Najim et al. 1998).

Preparation of the tested chemical agents

Sodium stibogluconate

A 30 ml vial was purchased from (Kolkata, India), patch no: (9p12004). It is containing 100 mg/ml antimony as active ingredient diluted down to 20 μg/ml. This drug was used as a control (Darrell et al. 2002).

The solvent (acetone)

A 1,000 ml bottle of acetone solution (100 %) was purchased from (Medex Company, patch no: (200-662-2). The solution of acetone (50 %) prepared by adding acetone: water (50:50 v/v). This solution was used as a solvent to the tested chemical agent (Chong et al. 2009).

Phenolic compounds

  1. Caffeic acid A 10 g crude powder of caffeic acid (CA) was purchased from Sigma Aldrich Company (Batch no. 21909058). Using acetone 50 % (as a solvent) and stirrer to prepare five different concentration of CA (10, 15, 20, 25, 30) mg/ml.

  2. Ferulic acid A 5 g crude powder of ferulic acid (FA) was purchased from Sigma Aldrich Company (Batch no. 76755-369). Using acetone 50 % (as a solvent) and stirrer to prepare five different concentration of FA (10, 15, 20, 25, 30) mg/ml.

  3. Syringic acid A 5 g crude powder of syringic acid (SA) is purchased from Sigma Aldrich Company (Batch no. 100989620). Using acetone 50 % (as a solvent) and stirrer to prepare five different concentration of SA (10, 15, 20, 25, 30) mg/ml.

  4. 4-Hydroxybenzoic acid A 50 g powder of 4-hydroxybenzoic acid (4-HBA) is purchased from Sigma Aldrich Company (Batch no. 1001034199). Using acetone 50 % (as a solvent) and stirrer to prepare five different concentration of 4-HBA (10, 15, 20, 25, 30) mg/ml.

Result

The effect of increasing concentration of

Phenolic compounds

After adding 10 mg of CA, FA, SA and 4-HBA the parasiticidal effect is significantly increased by a mean (24 × 104, 16 × 104, 16 × 104, 13 × 104 promastigote/ml) respectively compared to baseline controls. After each successive increase in concentration the parasiticidal effect of CA, FA, SA and 4-HBA is significantly increased to reach a maximum increase by a mean (53 × 104, 50 × 104, 44 × 104, 48 × 104 promastigote/ml) respectively compared to baseline control (5–6 × 104 promastigote/ml). The Cohen’s d effect size of CA, FA, SA, and 4-HBA is increased from (4.4, 5.7, 6.7, 3.2) respectively after 10 mg concentration to reach a maximum of CA (10.8), FA (9.6), SA (10.5), 4-HBA (5.2) after the highest concentration of 30 mg. The concentration of added CA, FA, SA, and 4-HBA showed a statistically significant (P < 0.001) positive strong linear correlation (r = 0.93, r = 0.96, r = 0.96, r = 0.89) respectively with parasiticidal effect. The regression model explaining the effect of CA, FA, SA, and 4-HBA concentration was statistically significant (P < 0.001) and able to explain (87, 93, 93, 78 %) respectively of variation in the dependent variable (parasiticidal effect). For each one mg increase in CA, FA, SA, and 4-HBA concentration the parasiticidal effect is expected to significantly increase by a mean of (1.69, 1.65, 1.47, 1.66) respectively.

Sodium stibogluconate

After adding 20 μg/ml of Sodium stibogluconate the parasiticidal effect is significantly increased by a mean of 79 × 104 promastigote/ml compared to baseline control (8 × 104 promastigote/ml). The Cohen’s d effect size reaches 26.3 after 20 μg/ml dose (Fig. 2).

Fig. 2.

Fig. 2

Open in a new tab

Line graph showing the linear trend of mean parasiticidal effect (rate of parasitic killing) with dosage of 5 additives

Comparing the effect of 5 additives

There was no obvious or statistically significant difference in mean parasiticidal effect at baseline control for the first 4 additives: CA, FA, SA and 4-HBA. The mean parasiticidal effect for 5th experiment baseline control (for sodium stibogluconate) was slightly higher (8 × 104 promastigote/ml) than the remaining 4 experiments (5–6 × 104 promastigote/ml). This difference would not affect the comparison of “changes” in parasiticidal effect after each specific concentration between different additives compared to baseline controls (Fig. 3).

Fig. 3.

Fig. 3

Open in a new tab

Dot diagram with error bars showing the mean (with its 95 % confidence interval) parasiticidal effect (rate of parasitic killing) at baseline (controls) between 5 study groups

After 10 mg concentration

There was a statistically significant (P < 0.001) difference in mean parasiticidal effect after 10 mg concentration between the 4 types of tested additives. The lowest parasiticidal effect was observed with 4-HBA (19 × 104 promastigote/ml). It will therefore be used as the reference (comparison) group. The strongest effect was observed with CA (29 × 104 promastigote/ml), resulting in a Cohen’s d effect size of 1.6, which is significantly higher compared to the reference group. Both ferulic and SA showed a slightly higher parasiticidal effect (21 × 104 promastigote/ml), which was not significantly higher than that of the reference group (4-HBA). The Cohen’s d effect size for both groups (ferulic and SA) was of small magnitude (0.5) (Fig. 4).

Fig. 4.

Fig. 4

Open in a new tab

Dot diagram with error bars showing the mean (with its 95 % confidence interval) parasiticidal effect (rate of parasitic killing) after a dosage of 10 mg of additive between 4 study groups

After 15 mg concentration

There was a statistically significant (P < 0.001) difference in mean parasiticidal effect after 15 mg concentration between the 4 types of tested additives. The lowest parasiticidal effect was observed with 4-HBA (27 × 104 promastigote/ml) (Fig. 5). It will therefore be used as the reference (comparison) group. The strongest effect was observed with CA (35 × 104 promastigote/ml), resulting in a Cohen’s d effect size of (1.5), which is significantly higher compared to the reference group. FA showed a slightly higher parasiticidal effect (28 × 104 promastigote/ml), which was not significantly higher than that of the reference group (4-HBA), SA have the same paraticidal effect of 4-HBA (27 × 104 promastigote/ml). The Cohen’s d effect size for (FA and SA) was of small magnitude (0.2, 0) respectively (Fig. 6).

Fig. 5.

Fig. 5

Open in a new tab

Dot diagram with error bars showing the mean change (with its 95 % confidence interval) in parasiticidal effect (rate of parasitic killing) after a dosage of 10 mg of additive compared to baseline (control) between 4 study groups

Fig. 6.

Fig. 6

Open in a new tab

Dot diagram with error bars showing the mean (with its 95 % confidence interval) parasiticidal effect (rate of parasitic killing) after a dosage of 15 mg of additive between 4 study groups

After 20 mg concentration

There was a statistically significant (P < 0.001) difference in mean parasiticidal effect after 20 mg concentration between the 4 types of tested additives. The lowest parasiticidal effect was observed with SA (35 × 104 promastigote/ml). It will therefore be used as the reference (comparison) group (Fig. 7). The strongest effect was observed with CA (43 × 104 promastigote/ml), resulting in a Cohen’s d effect size of (1.3), which is significantly higher compared to the reference group. Both Ferulic and 4-HBA showed a slightly higher parasiticidal effect (36 × 104 promastigote/ml), which was not significantly higher than that of the reference group (SA). The Cohen’s d effect size for both FA (0.3) and for 4-HBA (0.1) (Fig. 8).

Fig. 7.

Fig. 7

Open in a new tab

Dot diagram with error bars showing the mean change (with its 95 % confidence interval) in parasiticidal effect (rate of parasitic killing) after a dosage of 15 mg of additive compared to baseline (control) between 4 study groups

Fig. 8.

Fig. 8

Open in a new tab

Dot diagram with error bars showing the mean (with its 95 % confidence interval) parasiticidal effect (rate of parasitic killing) after a dosage of 20 mg of additive between 4 study groups

After 25 mg concentration

There was not a statistically significant (P < 0.08) difference in mean parasiticidal effect after 25 mg dose between the 4 types of tested additives. The lowest parasiticidal effect was observed with SA (43 × 104 promastigote/ml). It will therefore be used as the reference (comparison) group (Fig. 9). The strongest effect was observed with CA (49 × 104 promastigote/ml), resulting in a Cohen’s d effect size of (1), which is significantly higher compared to the reference group. Both Ferulic and 4-HBA showed a slightly higher parasiticidal effect (46 × 104 promastigote/ml), which was not significantly higher than that of the reference group (SA). The Cohen’s d effect size for ferulic and 4-HBA was of small magnitude (0.5, 0.3) respectively (Fig. 10).

Fig. 9.

Fig. 9

Open in a new tab

Dot diagram with error bars showing the mean change (with its 95 % confidence interval) in parasiticidal effect (rate of parasitic killing) after a dosage of 20 mg of additive compared to baseline (control) between 4 study groups

Fig. 10.

Fig. 10

Open in a new tab

Dot diagram with error bars showing the mean (with its 95 % confidence interval) parasiticidal effect (rate of parasitic killing) after a dosage of 25 mg of additive between 4 study groups

After 30 mg concentration

There was a statistically significant (P < 0.003) difference in mean parasiticidal effect after 30 mg dose between the 4 types of tested additives. The lowest parasiticidal effect was observed with SA (49 × 104 promastigote/ml) (Fig. 11). It will therefore be used as the reference (comparison) group. The strongest effect was observed with CA (58 × 104 promastigote/ml), resulting in a Cohen’s d effect size of (1.5), which is significantly higher compared to the reference group. Both Ferulic and 4-HBA showed a slightly higher parasiticidal effect (54 × 104 promastigote/ml), which was not significantly higher than that of the reference group (SA). The Cohen’s d effect size for ferulic and 4-HBA was of small magnitude (0.8, 0.5) respectively (Fig. 12).

Fig. 11.

Fig. 11

Open in a new tab

Dot diagram with error bars showing the mean change (with its 95 % confidence interval) in parasiticidal effect (rate of parasitic killing) after a dosage of 25 mg of additive compared to baseline (control) between 4 study groups

Fig. 12.

Fig. 12

Open in a new tab

Dot diagram with error bars showing the mean (with its 95 % confidence interval) parasiticidal effect (rate of parasitic killing) after a dosage of 30 mg of additive between 4 study groups

Discussion

Leishmaniases are a group of disease caused by Leishmania species, the disease is considered as a major public health problem in 82 countries in the world causing morbidity and mortality (WHO 1990).

There is a general lack of effective and in expensive chemotherapeutic agents for the treatment of leishmaniasis. Although trivalent antimonial like potassium antimonyl tartrate and pentavalent antimonial drugs are the first-line treatment for this disease, with amphotericin B and pentamidine being used as alternative drugs, all of these have serious side effects and resistance has become a challeng problem. Therefore, new drugs are urgently required, natural products have potential in the search for new and selective agents for the treatment of important tropical diseases caused by protozoans (Wright and Phillipson 1990). In the vast areas of the world, modern drugs are simply not available or if they are available they often prove to be too expensive. The majority of drugs active against infectious agents are in fact derived from natural products (Geoffrey 1996). Different plants of medicinal value are used traditionally worldwide for the treatment of leishmaniasis (Khalid et al. 2005). In the literature there are several reports on the activity of a variety of crude natural extracts, especially from plants collected in tropical zones against leishmania species (Fournet and Barrios 1994; Fournet et al. 1996; Akendengue et al. 1999; Weniger et al. 2001; Fournet and Munoz 2002). The parasite cultured in RPMI 1640 medium and counted the number of promastigote daily to determine the logarithmic growth phase, the fourth day (96 h) represent the logarithmic growth phase and the numbers reach to (4 × 106 promastigote/ml) (Fig. 13).

Fig. 13.

Fig. 13

Open in a new tab

Dot diagram with error bars showing the mean change (with its 95 % confidence interval) in parasiticidal effect (rate of parasitic killing) after a dosage of 30 mg of additive compared to baseline (control) between 4 study groups

The study of Mobarak (2008) shows the number of CL growing in RPMI 1640 medium reach to (2.8 × 106 promastigote/ml) in logarithmic growth phase. The study of Ibrahim et al. demonstrate the number of CL and VL growing in RPMI 1,640 medium reach to (5 × 107 promastigote/ml) in logarithmic growth phase (Ibrahim et al. 1994). Another study by Steiger and Steiger, demonstrate the number of Leishmania donovani and Leishmania braziliensis reach to (4 × 107 promastigote/ml) in logarithmic growth phase (Steiger and Steiger 1977).

The effect of phenolic compounds on Leishmania parasite viability is not yet known, and, to date, no other out comes studies have been announced.

It is observed that the four tested agents (CA, FA, SA and 4-HBA) have different parasiticidal effect depending on the difference in concentration after duration 72 h. The parasiticidal effect of each agent is tested according to five different concentrations (10, 15, 20, 25, 30 mg/ml).

This study demonstrates that CA has a strong parasiticidal activity depending on the variation in concentration after duration of 72 h. This study showed the mean of parasiticidal effect of CA at 10 mg/ml is (24 × 104 promastigote/ml) as compared to baseline control. After each successive increase in concentration the parasiticidal effect is increased to reach a mean (30 × 104 promastigote/ml) in a concentration 15 mg/ml. The parasiticidal effect more increased when adding (20, 25, 30 mg/ml) to reach a mean (38 × 104, 44 × 104, 53 × 104 promastigote/ml) respectively. These observations are in consistence with the study of Chong et al., reported that CA inhibits the radial growth of Ganoderma Boninense (Chong et al. 2009). Another study by Ismaiel and Pierson investigated the effect of CA against Clostridium botulinum, they state that CA reduces the viability of Clostridium spores (Ismaiel and Pierson 1990). The antimicrobial effect of CA may be attributed to the interaction with metal ions such as iron which is essential for microbial growth (Kontoghiorghes et al. 1986). Furthermore CA could cause change in the structure of cytoplasmic proteins that inhibit cell division (Cowan 1999). In another report, CA was also found inhibiting the growth of sweet potato pathogenic fungi, inhibitory activity reported and suggests high periderm CA levels contribute to the storage root defense chemistry of some sweet potato genotypes (Harrison et al. 2003).

In this study the effect of FA on the viability of Leishmania tropica promastigote are investigated and showed a significant impact on their number, an increase in FA concentration has a major effect on L. tropica promastigotes viability. This study demonstrated that the parasiticidal effect of FA at (10 mg/ml) is (16 × 104 promastigote/ml) compared to baseline control and the parasiticidal rate increased after increasing in concentration (15, 20, 25, 30 mg/ml) to reach a mean (23 × 104, 31 × 104, 42 × 104, 50 × 104 promastigote/ml) respectively. In this study FA is considered the best parasiticidal agent after CA in comparison to the other tested agents. This study is consistency with the study of Felaih et al. (2011), demonstrated in vitro study that phenolic compounds (CA and FA) have a fatal effect on the viability of protoscolices under different concentration and different durations (Felaih et al. 2011). The study of Aziz, which states that FA is able to inhibit growth of seed-born fungi, the possible explanation of the antimicrobial effect of FA is the ability of FA to interrupt the activity of the enzyme that responsible for DNA replication such as topoisomerase (Aziz et al. 1998). Another study by Franke et al., states that FA inhibits the respiratory syncycial virus reproduction (Natalia 2013). The study of Gardjeva et al. (2007), reported that FA has bactericidal activity against streptococcus beta-haemolyticus.

In this study showed the parasiticidal effect of SA at (10 mg/ml) is (16 × 104 promastigote/ml) compared to baseline control. The parasiticidal effect increased after increase in concentration to reach in a mean (22 × 104 promastigote/ml) at a concentration (15 mg/ml) and in a concentration (20, 25, 30 mg/ml) the parasiticidal effect reach to a mean (30 × 104, 38 × 104, 44 × 104 promastigote/ml) respectively. This study demonstrates that SA has a weak parasiticidal activity in comparison to the other tested agents.

This study is consistence with the experiment conducted by Chong et al. (2009), SA was found to be very fungi toxic to Ganoderma boninense, when the concentration was increased the pathogen was fully inhibited. Many works on plants have demonstrated the fungi toxicity effect of SA, in resistance raspberry to fungus Didymella, SA was found accumulated in the bordering zone of lesion forming a barrier to the fungus (Kozlowska and Krzywanski 1994). In sugar cane plant, which is highly resistant to smut disease showed a major accumulation pattern of SA when interact with the pathogen (De Armas et al. 2007).

In this study the effect of 4-HBA on the viability of Leishmania tropica promastigotes were investigated an increase in 4-HBA concentration has a major effect on Leishmania tropica promastigotes viability. This study demonstrated the mean at (10 mg/ml) is (13 × 104 promastigote/ml) as compared to baseline control. The parasiticidal effect increased as the concentration is varying (15, 20, 25, 30 mg/ml) to reach a mean (22 × 104, 31 × 104, 41 × 104, 48 × 104 promastigote/ml) respectively. In this study 4-HBA has a best paraciticidal activity after CA and FA in a comparison to other tested chemical agents. Cho et al. (1998), showed the role of 4-HBA in rice hull that have antimicrobial effect against various microorganisms, most of the gram-positive and some gram-negative bacteria. A study by Chong et al., reported that the synergy effect of SA, CA and 4-HBA against Ganoderma boninense, either syringic with 4-HBA or caffeic with 4-HBA or syringic with CA or the combination of all the three phenolics totally inhibited the growth of Ganoderma Boninense, the pathogen failed to grow in a combination of a high concentration of all the three phenolics but the combination of lower concentration of the phenolics which poorly inhibited the growth of the pathogen, but a better performance were shown in a combination of CA and 4-HBA in a high concentration (Chong et al. 2009). Maddox et al. (2010), observed that phenolic compounds that have antibacterial activity against Xylella fastidiosa.

Winkelhausen et al. (2005), demonstrated that the phenolic compounds derived from olive pomace inhibited the growth of fungi (Alternaria solani, Botrytis cinerea and Fusarium culmorum). Different researchers have also reported that phenolic compounds from different plant sources could inhibit various pathogens (Nychas 1995; Smid and Gorris 1999; Prashanth et al. 2001; Kim et al. 2005). The antimicrobial activities of phenolic compounds may involve multiple modes of action. For example, essential oils degrade the cell wall, interact with the composition and disrupt cytoplasmic membrane (Sikkema et al. 1994; Helander et al. 1998; Ultee et al. 1999; Lambert et al. 2001),damage membrane protein, interfere with membrane integrated enzymes (Raccach 1984), cause leakage of cellular components, coagulate cytoplasm, deplete the proton motive force, change fatty acid and phospholipid constituents, impair enzymatic mechanisms for energy production and metabolism, alter nutrient uptake and electron transport (Taniguchi et al. 1988), influence the synthesis of DNA and RNA and destroy protein translocation and the function of the mitochondrion in eukaryotes (Nychas 1995; Raccach 1984). All of these mechanisms are not separate targets; some are affected as a consequence of another mechanism being targeted. The mode of action of antimicrobial agents also depends on the type of microorganisms and is mainly related to their cell wall structure and the outer membrane arrangement (Kalemba and Kunicka 2003; Burt 2004).

References

  1. Akendengue B, Ngou-Milama E, Laurens A, Hocquemiller R. Recent advances in the fight against leishmaniasis with natural products. Parasite. 1999;6(3–8):20. doi: 10.1051/parasite/1999061003. [DOI] [PubMed] [Google Scholar]
  2. Aziz NH, Farag SE, Mousa LAA, Abu-Zaid MA. Comparative antibacterial and antifungal effects of some phenolic compounds. Microbios. 1998;93:43–54. [PubMed] [Google Scholar]
  3. Berman JD. Treatment of New World cutaneous and mucosal leishmaniases. Clin Dermatol. 1996;14:519–522. doi: 10.1016/0738-081X(96)00048-X. [DOI] [PubMed] [Google Scholar]
  4. Berman JD. Human leishmaniasis: clinical, diagnostic, and chemotherapeutic developments in the last 10 years. Clin Infect Dis. 1997;24:684–703. doi: 10.1093/clind/24.4.684. [DOI] [PubMed] [Google Scholar]
  5. Boelaert M, Le-ray D, Van-Der SP. How better drugs could change kala-azar control. Lessons from a cost-effectiveness analysis. Trop Med Int Health. 2002;7:955–959. doi: 10.1046/j.1365-3156.2002.00959.x. [DOI] [PubMed] [Google Scholar]
  6. Burt S. Essential oils: their antibacterial properties and potential applications in foods—a review. Int J Food Microbiol. 2004;94:223–253. doi: 10.1016/j.ijfoodmicro.2004.03.022. [DOI] [PubMed] [Google Scholar]
  7. Carrio J, Riera C, Gallego M, Ribera E, Portus M. In vitro susceptibility of Leishmania infantum to meglumine antimoniate in isolates from repeated leishmaniasis episodes in HIV-coinfected patients. J Antimicrob Chemother. 2001;47:120–121. doi: 10.1093/jac/47.1.120. [DOI] [PubMed] [Google Scholar]
  8. Cho J, Moon J, Seong K, Park K. Antimicrobial activity of 4-hydroxybenzoic acid and trans 4-hydroxycinnamic acid isolated and identified from Rice Hull. Biosci Biotechnol Biochem. 1998;62(11):2273–2276. doi: 10.1271/bbb.62.2273. [DOI] [PubMed] [Google Scholar]
  9. Chong KP, Stephen R, Markus A. In vitro antimicrobial activity and fungitoxicity of syringic acid, caffeic acid and 4-hydroxybenzoic acid against Ganoderma Boninense. JAS. 2009;1(2):8. [Google Scholar]
  10. Cowan MM. Plant products as antimicrobial agent. Clin Microbiol Rev. 1999;12:564. doi: 10.1128/cmr.12.4.564. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Darrell R, Gary J, Robert L, Dawn M, Robert A, Keith R et al (2002) Mosby’s drug consult, section III, drug information. Copywrite © 2002, Mosby Inc, p 674
  12. Dawson RMC, Elliot DC, Elliot WH, Jons KM. Data for biochemical research. 2. Oxford: Clareden Press; 1969. p. 508. [Google Scholar]
  13. De Armas R, Santiago R, Legaz ME, Vicente C. Levels of phenolic compounds and enzyme activity can be used to screen for resistance of sugarcane to smut (Ustilago scitaminea) Australas Plant Pathol. 2007;36(1):32–38. doi: 10.1071/AP06077. [DOI] [Google Scholar]
  14. Desjeux P. Leishmaniasis: public health aspects and control. Clin Dermatol. 1996;14:417–423. doi: 10.1016/0738-081X(96)00057-0. [DOI] [PubMed] [Google Scholar]
  15. Ephros M, Waldman E, Zilberstein D. Pentostam induces resistance to antimony and the preservative chlorocresol in Leishmania donovani promastigotes and axenically grown amastigotes. Antimicrob Agents Chemother. 1997;41:1064–1068. doi: 10.1128/aac.41.5.1064. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Felaih HI, Alshammary MN, Mohammad BI (2011) Effect of phenolic compounds and terpenoid on the viability of Echinococcus granulosus protoscolices: a comparative in vitro study. Master thesis. Al-Qadissiyah University, College of Medicine
  17. Fournet A, Barrios AA. Leishmanicidal and trypanocidal activities of Bolivian medicinal plants. J Ethnopharmacol. 1994;41:19–37. doi: 10.1016/0378-8741(94)90054-X. [DOI] [PubMed] [Google Scholar]
  18. Fournet A, Munoz V. Natural products as trypanocidal, antileishmanial and antimalarial drugs. Curr Top Med Chem. 2002;2:1215–1237. doi: 10.2174/1568026023393011. [DOI] [PubMed] [Google Scholar]
  19. Fournet A, Ferreira ME, Rojas-De-Arias A, Torres-De-Ortiz S, Fuentes S, Nakayama H, Schinini A, Hocquemiller R. In vivo efficacy of oral and intralesional administration of 2-substituted quinolines in experimental treatment of world cutaneous leishmaniasis caused by Leishmania amazonensis. Antimicrob Agents Chemother. 1996;40:2447–2451. doi: 10.1128/aac.40.11.2447. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Gardjeva PA, Dimitrova SZ, Kostadinov ID, Murdjeva MD, Peyehe LP. A study of chemical composition and antimicrobial activity of Bulgarian propolis. Folia Med. 2007;49:63–69. [PubMed] [Google Scholar]
  21. Geoffrey CK. Medicinal plants and the control of protozoal disease, with particular reference malaria. Trans R Soc Trop Med Hyg. 1996;90:605–609. doi: 10.1016/S0035-9203(96)90404-6. [DOI] [PubMed] [Google Scholar]
  22. Harrison HF, Peterson JK, Snook ME, Bohac JR, Jackson DM. Quantity and potential biological activity of caffeic acid in sweet potato storage root periderm. J Agric Food Chem. 2003;51(10):2943–2948. doi: 10.1021/jf0211229. [DOI] [PubMed] [Google Scholar]
  23. Helander IM, Alakomi H-L, Latva-Kala K, Mattila-Sandholm T, Pol I, Smid EJ, Gorris LGM, Von Wright A. Characterization of the action of selected essential oil components on Gram-negative bacteria. J Agric Food Chem. 1998;46:3590–3595. doi: 10.1021/jf980154m. [DOI] [Google Scholar]
  24. Ibrahim ME, Hag-Ali M, El-Hassan AM, Theander TG, Kharazmi A. Leishmania resistant to sodium stibogluconate: drug-associated macrophage-dependent killing. Parasitol Res. 1994;80:569–574. doi: 10.1007/BF00933004. [DOI] [PubMed] [Google Scholar]
  25. Ismaiel AA, Pierson MD. Inhibition of germination, out growth and vegetative growth of clostridium botulinum by spice oils. J Food Prot. 1990;53:755–758. doi: 10.4315/0362-028X-53.9.755. [DOI] [PubMed] [Google Scholar]
  26. Kagan IG, Norman L. Manual of clinical microbiology. Washington: American Society for Microbiology; 1970. pp. 453–486. [Google Scholar]
  27. Kalemba D, Kunicka A. Antibacterial and antifungal properties of essential oils. Curr Med Chem. 2003;10:813–829. doi: 10.2174/0929867033457719. [DOI] [PubMed] [Google Scholar]
  28. Khalid FA, Abdalla NM, Mohomed HO, Toum AM, Magzoub MMA, Ali MS. In vitro assessment of anti-cutaneous leishmaniasis activity of some sudanese plants. Türkiye Parazitol Derg. 2005;29(1):3–6. [PubMed] [Google Scholar]
  29. Killick-Kendrick R. Phlebotomine vectors of the leishmaniases. Med Vet Entomol. 1990;4(1):1–24. doi: 10.1111/j.1365-2915.1990.tb00255.x. [DOI] [PubMed] [Google Scholar]
  30. Kim YS, Hwang CS, Shin DH. Volatile constituents from the leaves of Polygonum cuspidatum S. et Z. and their anti-bacterial activities. Food Microbiol. 2005;22:139–144. doi: 10.1016/j.fm.2004.01.016. [DOI] [Google Scholar]
  31. Kontoghiorghes GJ, Jackson MJ, Lunec J. In vitro screening of iron chelators using models of free radical damage. Free Radic Res Commun. 1986;2:115–124. doi: 10.3109/10715768609088062. [DOI] [PubMed] [Google Scholar]
  32. Kozlowska M, Krzywanski Z. The possible role of phenolic compounds in red raspberry resistance to Didymella applanata (Niessl) Sacc. Acta Hortic. 1994;381:671–674. doi: 10.17660/ActaHortic.1994.381.95. [DOI] [Google Scholar]
  33. Lambert RJW, Skandamis PN, Coote P, Nychas GJE. A study of the minimum inhibitory concentration and mode of action of oregano essential oil, thymol and carvacrol. J Appl Microbiol. 2001;91:453–462. doi: 10.1046/j.1365-2672.2001.01428.x. [DOI] [PubMed] [Google Scholar]
  34. Lira R, Sundar S, Makharia A, Kenney R, Gam A, Sacks D. Evidence that the high incidence of treatment failures in Indian kala-azar is due to the emergence of antimony-resistant strains of Leishmania donovani. J Infect Dis. 1999;180:564–567. doi: 10.1086/314896. [DOI] [PubMed] [Google Scholar]
  35. Maddox CE, Laur LM, Tian Li. Antibacterial activity of phenolic compounds against the pathogen of Xylella fastidiosa. Curr Microb. 2010;60(1):53–58. doi: 10.1007/s00284-009-9501-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Mobarak HA (2008). Isolation and cultivation of cutaneous leishmania parasite by using different cultures media. Master thesis. University of Kufa, College of Medicine, p 22
  37. Najim RA, Sharquie KE, Farjo IB. Zinc sulfate in the treatment of cutaneous leishmaniasis an in vitro and animal study. Men Inst Oswaldo Ciuz. 1998;93:831–837. doi: 10.1590/S0074-02761998000600025. [DOI] [PubMed] [Google Scholar]
  38. Natalia G (2013) Planktonics or biofilms infections? AJBM 1(1):6–8
  39. Nychas GJE. Natural antimicrobials from plants. In: Gould GW, editor. New methods of food preservation. London: Blackie Academic; 1995. pp. 58–89. [Google Scholar]
  40. Osman OF, Kager PA, Oskam L. Leishmaniasis in the Sudan: a literature review with emphasis on clinical aspects. Trop Med Int Health. 2000;5:553–571. doi: 10.1046/j.1365-3156.2000.00598.x. [DOI] [PubMed] [Google Scholar]
  41. Prashanth D, Asha MK, Amit A. Antibacterial activity of Punica granatum. Fitoterapia. 2001;72:171–173. doi: 10.1016/S0367-326X(00)00270-7. [DOI] [PubMed] [Google Scholar]
  42. Raccach M. The antimicrobial activity of phenolic antioxidants in food: a review. J Food Saf. 1984;6:141–170. doi: 10.1111/j.1745-4565.1984.tb00479.x. [DOI] [Google Scholar]
  43. Ranga VS, Julia LH (1997) Immunology method manual, chapter 29.2, vol. 4. In: HIV growth, measurement and neutralization, pp 1963–1964
  44. Ready PD. Leishmaniasis emergence and climate change. In: de la Roque S, editor. Climate change: the impact on the epidemiology and control of animal diseases. Paris: Revue Scientifique et Technique (International Office Epizootics); 2008. pp. 399–412. [Google Scholar]
  45. Roberts MTM. Current understandings on the immunology of leishmaniasis and recent developments in prevention and treatment. Br Med Bull. 2006;75(76):115. doi: 10.1093/bmb/ldl003. [DOI] [PubMed] [Google Scholar]
  46. Sikkema J, De Bont JAM, Poolman B. Interactions of cyclic hydrocarbons with biological membranes. J Biol Chem. 1994;269:8022–8028. [PubMed] [Google Scholar]
  47. Smid EJ, Gorris LGM. Natural antimicrobials for food preservation. In: Rahman MS, editor. Handbook of food preservation. New York: Marcel Dekker; 1999. pp. 285–308. [Google Scholar]
  48. Steiger RF, Steiger E. Cultivation of Leishmania donovani and Leishmania braziliensis in defined media: nutritional requirements. J. Protozool. 1977;24(3):441–443. doi: 10.1111/j.1550-7408.1977.tb04771.x. [DOI] [PubMed] [Google Scholar]
  49. Takahashi M, Fuchino H, Sekita S, Satake M. In vitro leishmanicidal activity of some scarce natural products. Phytother Res. 2004;18:573–578. doi: 10.1002/ptr.1502. [DOI] [PubMed] [Google Scholar]
  50. Taniguchi M, Yano Y, Tada E, Ikenishi K, Ol S, Haraguchi H, Hashimoto K, Kubo I. Mode of action of polygodial: an antifungal sesquiterpene dialdehyde. Agric Biol Chem. 1988;52:1409–1414. doi: 10.1271/bbb1961.52.1409. [DOI] [Google Scholar]
  51. Thakur CP. Socio-economics of visceral leishmaniasis in Bihar (India) Trans R Soc Trop Med Hyg. 2000;94:156–157. doi: 10.1016/S0035-9203(00)90255-4. [DOI] [PubMed] [Google Scholar]
  52. Ultee A, Kets EPW, Smid EJ. Mechanisms of action of carvacrol on the food-borne pathogen Bacillus cereus. Appl Environ Microbiol. 1999;65:4606–4610. doi: 10.1128/aem.65.10.4606-4610.1999. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Waterman PG, Mole S. Analysis of phenolic plant metabolites. Oxford: Blackwell Scientific Publications; 1994. [Google Scholar]
  54. Weniger B, Robledo S, Arango GJ, Deharo E, Aragon R, Munoz V, Callapa J, Lobstein A, Anton R. Antiprotozoal activities of Colombian plants. J Ethnopharmacol. 2001;78:193–200. doi: 10.1016/S0378-8741(01)00346-4. [DOI] [PubMed] [Google Scholar]
  55. WHO (1990) Control of the leishmaniases. Report of WHO expert committee, technical series, no 793 [PubMed]
  56. WHO (2007) Sixtieth world health assembly. Provisional agenda item. 12, 3, pp 1–5
  57. Winkelhausen ER, Pospiech R, Laufenberg G. Antifungal activity of phenolic compounds extracted from dried olive pomace. Bull Chem Technol Maced. 2005;24(1):41–46. [Google Scholar]
  58. Wright CW, Phillipson JD. Natural products and the development of selective antiprotozoal drugs. Phytother Res. 1990;4:127–139. doi: 10.1002/ptr.2650040402. [DOI] [Google Scholar]

Articles from Journal of Parasitic Diseases: Official Organ of the Indian Society for Parasitology are provided here courtesy of Springer

RESOURCES