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Antisalmonellal Activity and Phytochemical Screening of the Various Parts of Cassia petersiana Bolle (Caesalpiniaceae)



Donatien Gatsing and Godwin I. Adoga
 
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ABSTRACT

Aqueous extracts of the various parts (i.e., flowers, leaves, barks and roots) of Cassia petersiana (Caesalpiniaceae) were tested for antibacterial activity against Salmonella typhi, Salmonella paratyphi A and Salmonella paratyphi B using both agar diffusion and broth dilution techniques. Phytochemical screening was also performed using standard methods. All the parts tested (at 30 mg mL-1) showed antibacterial activity against all the three bacteria strains, with the diameters of zones of inhibition ranging form 7 to 19 mm. The leaves showed the highest activity, followed by the flowers, the roots and then the barks. The MIC (Minimum Inhibitory Concentration) and MBC (Minimum Bacterial Concentration) values of the leaf extract (i.e., the most potent extract) were 1.0 and 8.0 mg mL-1, respectively, against all the three bacterial strains. Phytochemical screening gave evidence of the presence of compounds of biological interest (i.e., alkaloids, flavonoids, cardiac glycosides, antraquinones, anthocyanins, polyphenols, triterpenes, steroids, saponins, tannins and phlobatannins) in the various part of Cassia pertersiana. Anthraquinones and phlobatannins were present only in the leaf extract. These results suggest that the leaf extract may be much more recommended to the users of C. petersiana parts in the treatment of typhoid and paratyphoid fevers.

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  How to cite this article:

Donatien Gatsing and Godwin I. Adoga, 2007. Antisalmonellal Activity and Phytochemical Screening of the Various Parts of Cassia petersiana Bolle (Caesalpiniaceae). Research Journal of Microbiology, 2: 876-880.

DOI: 10.17311/jm.2007.876.880

URL: https://scialert.net/abstract/?doi=jm.2007.876.880

INTRODUCTION

Cassia petersiana Bolle (Caesalpiniaceae) is a tree commonly found growing on sandy soils up to 12 m high (Djemgou et al., 2006). It has pinnate green leaves and yellow flowers and is usually grown for ornamental purposes. The roots of the plant are used as a treatment for coughs, colds, syphilis and stomachache. It is also used as an anthelmentic (Gatsing et al., 2007). Biologically active compounds have been detected within species of the family Caesalpiniaceae; extracts of C. petersiana have been found to exhibit in vitro antimalarial activity against the multi-drug-resistant strain of Plasmodium falciparum (Connelly et al., 1996). In Southern Africa, the leaf is used as a febribuge and as a cure for skin diseases (Djemgou et al., 2006). Two chomones and other constituents from C. petersiana have been reported to exhibit antitumour and immunostimulatory activities (Djemgou et al., 2006). In northern Nigeria, various parts (flowers, leaves, barks, roots) of C. petersiana are boiled in water and used as remedies for typhoid fever.

Typhoid, paratyphoid A and paratyphoid B fevers are caused by Salmonella typhi, Salmonella paratyphi A and Salmonella paratyphi B, respectively (Gatsing et al., 2006). Typhoid fever continues to be a marked public health problem in developing countries in general and in sub-Saharan Africa in particular, where it is endemic (Gatsing et al., 2006). Conventional antisalmonellal drugs are becoming more and more unavailable to the common man in Africa due to increased cost (Gatsing et al., 2003). Moreover, the typhoid causative organism, Salmonella typhi, has rapidly gained resistance to all three first line antimicrobials (ampicillin, chloramphenicol and cotrimoxazole) and also to the previously efficacious drugs like ciprofloxacin (Madhulika et al., 2004).

In a continuation of our search for therapeutic agents from natural sources with potential for the treatment of typhoid and paratyphoid fevers (Aliyu et al., 2002; Teponno et al., 2006; Gatsing et al., 2003, 2006, 2007), various parts (i.e., flowers, leaves, barks and roots) of C. petersiana were selected and tested against Salmonella typhi, Salmonella paratyphi A and Salmonella paratyphi B. this was with a view to ascertaining and assessing their traditionally claimed antityphoid properties. The phytochemical compounds contained in the various parts of this plant were also investigated.

MATERIALS AND METHODS

Plant Sample
The flowers, leaves, barks and roots of Cassia petersiana were collected from the garden of the University of Jos main campus, Jos, Nigeria, in February 2003. The plant sample was identified by Mr. Kareem I.A., a plant taxonomist at the Federal College of Forestry, Jos, Nigeria and was further authenticated by Dr. Onana at the Cameroon National Herbarium, Yaoundé, where a voucher specimen (N° 6494/SFR/Cam) is deposited.

Test Bacteria and Culture Media
The bacteria strains, including Salmonella typhi, Salmonella paratyphi A and Salmonella paratyphi B, were obtained from the Medical Bacteriology Laboratory of the Pasteur Centre, Yaoundé, Cameroon. The culture media used namely Salmonella-Shigella agar (SS agar) and Selenite broth, were obtained from International Diagnostics Group PLC, UK.

Preparation of Extracts
The flowers, leaves, barks and roots of C. Petersiana were air-dried and pulverized. Each powder sample (500 g) was soaked in 2.5 L of water, boiled for 15 min and filtered. The filtrate was concentrated in a drying oven at 45°C.

Antibacterial Assay
The antibacterial activity was determined using both agar diffusion and broth dilution techniques as previously described by Gatsing et al. (2006).

The diffusion sensitivity test was performed using well method and used to screen the various parts of Cassia petersiana for their antisalmonellal activity. Each extract was tested at the concentration of 30 mg mL-1. The extract (i.e., the part of the plant) showing the highest diameter of inhibition was further studied and the MIC (Minimum Inhibitory Concentration) and MBC (Minimum Bactericidal Concentration) were determined using dilution sensitivity test. The extract was 2-fold serially diluted and the concentration ranges were 0.125-16 mg mL-1. Chloramphenicol was used as the standard antimicrobial and the concentration ranges were 0.125-16 μg mL-1.

Phytochemical Screening
The phytochemical screening was done using standard methods described by Harborne (1973) and Odebiyi and Sofowora (1978). The plant sample was screened for the following classes of compounds; alkaloids, flavonoids, cardiac glycosides, anthraquinones, anthocyanins, polyphenols, triterpenes, steroids, saponins, tannins and phlobatannins.

RESULTS AND DISCUSSION

Four different parts (leaf, flower, bark and root) of Cassia petersiana were extracted in water and screened for antisalmonellal activity using both agar diffusion and broth dilution methods. The extracts of the various parts, tested at 30 mg mL-1, showed antibacterial activity against all the three bacteria species with the diameters of zones of inhibition ranging from 7 to 19 mm (Table 1). The leaf extract was the most active, with diameters of inhibition of 13-16 mm against S. typhi, 16-17 mm against S. paratyphi A and 17-19 mm against S. paratyphi B. For the flower extract, the diameters of inhibition were 10-11 mm against S. typhi and S. paratyphi B and 12-14 mm against S. paratyphi A. For the root extract, the diameters of inhibition were 9-11 mm against S. typhi, 10-11 mm against S. paratyphi A and 11-13 mm against S. paratyphi B. The bark extract showed the lowest activity and its diameters of inhibition were 7-8 mm against S. typhi, 7-9 mm against S. paratyphi A and 9-10 mm against S. paratyphi B. Chloramphenicol (0.1 mg mL-1), used as the standard, showed the diameters of 27-28 mm, 25-27 mm and 26-28 mm against S. typhi, S. paratyphi A and S. paratyphi B, respectively.

The antisalmonellal activities showed by the various parts of C. petersiana are in line with the findings of Connelly et al. (1996) which showed in vitro antimalarial activity of extracts of C. petersiana against the multi-drug-resistant strain of Plasmodium falciparum.

The leaf extract, which showed the highest antibacterial activity against all the three bacteria strains used, was further studied using broth dilution technique and the following result were obtained; the MIC and MBC values were 1.0 and 8.0 mg mL-1, respectively, against all the three bacteria tested. For chloramphenicol the MIC and MBC values were 2.0 and 16.0 μg mL-1, respectively, against the same bacteria species (Table 2).

Antibacterial substances are considered as bactericidal agents when the ratio MBC/MIC≤4 and bacteriostatic agents when the ratio MBC/MIC>4 (Carbonnelle et al., 1987; Gatsing et al., 2006, 2007). For the aqueous leaf extract, the ratio MBC/MIC>4, suggesting that it may be classified as bacteriostatic agent.

From the result of the diffusion sensitivity test, it is observed that the leaf showed the highest activity, followed by the flower, the root and then the bark. These data confirm their traditionally claimed antityphoid properties. Moreover, these results indicate that the antisalmonellal principles in C. petersiana are more concentrated in the leaf than in any other part of the plant, suggesting that the leaf extract may be used in the treatment of typhoid and paratyphoid fevers with more favourable outcome.

The various parts of Cassia petersiana were screened for some classes of phytochemical compounds (Table 3). Polyphenols, saponins and tannins were found in all the parts of the plant. Alkaloids were present only in the bark, whereas flavonoids were found in all but the leaf. Cardiac glycosides were present in the bark and the root, while anthraquinones and phlobatannins were found only in the leaf. Anthocyanins were present in the flowers and the leaf, whereas triterpenes were found in the bark and the root. Steroids were found in all but the flower.

Table 1: Diameters of inhibition of the bacteria by the aqueous extracts of the various parts of Cassia petersiana
Image for - Antisalmonellal Activity and Phytochemical Screening of the Various Parts of Cassia petersiana Bolle (Caesalpiniaceae)

Table 2: MIC and MBC values of the aqueous extract of the leaves of Cassia petersiana
Image for - Antisalmonellal Activity and Phytochemical Screening of the Various Parts of Cassia petersiana Bolle (Caesalpiniaceae)
Key: MIC = Minimum Inhibitory Concentration; MBC = Minimum Bactericidal Concentration

Table 3: Some classes of phytochemical compounds present in Cassia petersiana
Image for - Antisalmonellal Activity and Phytochemical Screening of the Various Parts of Cassia petersiana Bolle (Caesalpiniaceae)
+: Present; –: Not present

The results of the phytochemical screening gave evidence of the presence of compounds of biological interest in the various parts of C. petersiana.

In effect, some flavonoids have shown several pharmacological activities among which antibacterial and antifungal activities (Leung, 1980). Some triterpenes have presented a protective function by participating in the fight against microbial attacks and as insectifuge (Harborne, 1973). Components belonging to the anthraquinone class (e.g., nordamnacanthal, damnacanthal and norindone) have exhibited high antimicrobial activities (Ali et al., 2000). Some alkaloids have shown antimicrobial activities against Gram positive and Gram negative bacteria (Faizi et al., 2003; Gonzaga et al., 2003). The tannins prevent the bacterial growth by precipitating their proteins (Fluck, 1976). Some saponins have presented antifungal activities (Etsuji et al., 1984).

The antibacterial activity of C. petersiana extracts may thus be due to the presence of alkaloids, flavonoids, triterpenes, tannins or saponins. Besides, the activity of the leaf extract may be enhanced by the presence of anthraquinones and/or phlobatannins.

The antisalmonellal activity of the aqueous leaf extract obtained in this work is greater than that of the CH2Cl2/MeOH (1:1) leaf extract obtained in a previous work (Gatsing et al., 2007), suggesting that the active principle(s) may be highly polar.

In the light of the foregoing, the leaf extract, which showed the highest antisalmonellal activity, may be much more recommended to the users of C. petersiana parts in the treatment of typhoid and paratyphoid fevers. However, further studies should be done to determine the possible side effects of this extract and to identify its active principle (s).

ACKNOWLEDGMENT

We wish to express our gratitude to Dr. (Mrs.) Fonkoua Marie-Christine, Medical Bacteriology Laboratory, Pasteur Centre, Yaoundé, Cameroon, for her co-operation.

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