Efficacy of Antimicrobial properties of Acalypha indica against Clinical isolates of human Pathogen

 

Desh Deepak Singh

Amity Institute of Biotechnology, Amity University Rajasthan, India-303002.

 

ABSTRACT:

Study was to investigates antimicrobial properties against clinical isolates of human pathogen, physiochemical and Phytochemical analysis of Acalypha indica. The aqueous, methanol, acetone and chloroform fractions of Acalypha indica were investigated for antimicrobial properties using pathogenic species of 18 bacteria and 8 fungi.  Efficacy of antimicrobial properties were shown by all fraction. Significant antimicrobial properties were observed by disc diffusion assay and Microbroth dilution assay against tested pathogen such as Staphylococcus epidermidis, Staphylococcus epidermidis, Streptococcus pyrogens, Salmonella typhi, Bacillus cereus, Escherichia coli, P. aeurigonosa, S. aureus, A. fumigatus, A. flavus, A. niger, C. albicans and C. tropicals.  The MICs by microbroth dilution assay was found to be ranged from 0.625 to 10.0 mg/ml in most of the fractions. Chloroform extract was found to be least active against the fungal pathogens. All the fractions of Acalypha indica were exhibited moderate to high activity against human pathogenic microbes; so that, identification of various extracts may be useful in novel antimicrobial compound having less toxicity with high sensitivity and specificity. 

 

 

 

INTRODUCTION:

Traditional natural products have been an important source of biologically active compounds for treating various disease conditions¹. The natural products are moderately cheap source of biological material having a vast variety of metabolites, primary or secondary, available in them for selecting the molecule of desired biological activity. Subsequently the beginning of development humans has been using herbs in contradiction of various pathogen². In the past few years, several antimicrobial agents have developed due to increase in the new spectrum of microbial infections and also the resistance against available drugs^3,4.

 

Extracts from the leaves of Acalypha indica exhibited various biological properties as itches, swellings, bilious fever, catarrh, eczema, tetanus, rheumatism and nematocidal activity has been reported^5,6. The leave of this plant is also used as emetic purgative and in joint pain⁷.

 

We have investigated the antimicrobial properties against pathogen by using water, methanol and chloroform extracts of Acalypha indica.

 

MATERIAL AND METHODS:

Plant collection:

Ariel parts of Acalypha indica were collected and fresh plant material was crushed by using a homogenizer. Plant powder was extracted in water, methanol, acetone and chloroform. All the extracts were named accordingly and the solvent was evaporated using rotavapour. The dried material was stored at 4⁰C till further use.

 

 

Microrganisms:

Clinical isolates along with standard bacterial strains of  bacteria (Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, Streptococcus pyrogens, Alcaligene fecalis, Salmonella typhi, Bacillu cereus, Bacillus subtilis, Micrococcus luteus, Micrococcus roseus, Streptococcus faecalis, Klebsiella pneumonia, Proteus vulgaris, Proteus mirabilis, Escherichia coli) and  fungi (Aspegillus  fumigatus, A. niger, A. flavus, Candida albicans and C. tropicals, Microsporum canis, Epidermophyton floccossum, Trichophyton rubrum) were used to find out the antibacterial activity of the extracts. For antifungal activity assays, standard strains of Aspergillus fumigatus, A. niger, A. flavus, Candida albicans and C. tropicals, were included in each test as recommended by National Committee for clinical laboratories standards (NCCLS). 

 

Micro-broth Dilution Assay (MDA):

The evaluations of susceptibility of all the MDA was carried out according to CLSI and EUCAST guideline^8-14. 

 

Disc Diffusion Assay (DDA):

DDA was performed according The evaluations of susceptibility of all the MDA was carried out according to CLSI and EUCAST guideline.^8-14

 

Phytochemicals screening:

The pre-liminary phytochemical screening was cried outby  saxena and saxena et al., 2012¹⁵ .

 

Qualitative analysis of inorganic elements:

Plant extract ash (500 mg) was analysed by Khandelwal, 2006¹⁶.

 

Quantitative analysis of phytochemicals:

Quantitative analysis was performed by using Senguttuvan et al., 2014¹⁷.

Histochemical analysis:

Histochemical analysis was performed by Gupta et al. 2006¹⁶.

 

RESULTS AND DISCUSSION:

Efficacy of antimicrobial properties of Acalypha indica was observed agaist all testeted human pathogen such as S. typhi, S. aeurigonosa, S. aureus, S. pyogenes, B. cereus, B. subtilis, M. luteus, S. faecalis, P. mirabilis and E. coli, A. fumigatus, A. flavus, A. niger, C. albicans and C. tropicals. 

 

The MICs by microbroth dilution assay was found to be ranged from 0.625 to 10.0 mg/ml in most of the fractions (Figure 1). Chloroform extract was found to be least active against the fungal pathogens. All the fractions of Acalypha indica were exhibited moderate to high activity as shown in figure 2 by DDA.  Proximate analysis of Acalypha indica were observed as shown in figure 3. Phytochemical analysis was also performed as shown in table 1in all extraction of Acalypha indica such as Steroids, Terpenoids, Flavonoids, Saponins, Tannins, Anthraquinone, Phlobatanins Polyphenol Protein Alkaloids Histochemical studies of Acalypha indica leaf powder (table 1). Qualitative analysis of elements in Acalypha indica was also performed as shown in table 2. The current antimicrobial agent challenging and resistant day by day against microbial infections, its needs to develop new antimicrobial agent for emerging and resistant pathogen. In this connections natural and bioactive product must use to serve world level requirements. Natural have advantages in drug discovery because of its significant antimicrobial activity and less toxicity. Therefore, author aimed to evaluate the antimicrobial activity against clinical isolates of human pathogen, physiochemical and Phytochemical analysis of Acalypha indica .

 

 

Figure:1 Antiicrobial activity of Acalypha indica (mg/ml) by MDA

 

 

Figure: 2 MIC of Acalypha indica L extracts against the microorganisms by disc diffusion assay.

 

 

Figure: 3 Proximate analysis

 

 

Table 1: Qualitative phytochemical analysis in Acalypha indica extract

S N	Extract	Carbohydrate	C. Glycosides	Steroids	Terpenoids	Flavonoids
1	Water	-	-	+	+	-
2	Methanol	+	+	+	-	+
3	Acetone	-	-	+	+	-
4	Chloroform	+	+	+	+	+
	Histochemical studies of Acalypha indica leaf powder	Cream in colour	Cream in colour	Violet to blue (or) Green	Orange	Yellow

 

Contiune Table 1

S N	Saponins	Tannins	Anthraquinone	Phlobatanins	Polyphenol	Protein	Alkaloids
1	+	+	+	+	-		-
2	+	-	-		-		+
3	+	+	+	+	+	-	+
4	+	+	+	+	+	+	+
	Yellow	Dark Blue to Black	Dark Blue green	Gary	Blue green/Red	Cream in colour	Reddish Brown

 

 

 

Table 2: Qualitative analysis of elements in Acalypha indica

S. No	Elements	Result
1.	Iron	-
2.	Aluminium	+
3.	Barium	+
4.	Calcium	+
5.	Chromium	+
6.	Copper	+
7.	Potassium	+
8.	Lithium	+
9.	Magnesium	+
10.	Zinc	+
11	Sodium	+
12	Nitrate	+
13	Phosphate	+
14	Sulphate	+
15	Chloride	+
Qualitative analysis of vitamins in Lantana indica L
1	Vitamins
2	A	-
3	C	+
4	D	-
4	E	+

 

CONCLUSION:

Most of the antimicrobial agent are resistant day by day against bacterial and fungal infection, therefore, its needs to develop new lead molecules for emerging and resistant infectious agent, Active molecule can be further fractionated from Acalypha indica to develop sensitive specific and less toxic antimicrobial agent. 

 

ACKNOWLEDGEMENTS:

The author gratefully acknowledges Dr. Vinod Kumar Tiwari, Department of Chemistry, Banaras Hindu university, Varanasi India; Prof. Amita Jain, Department of Microbiology King George’s Medical University, Lucknow, India; Dr. Rambir Singh, Institute of Biomedical sciences, Bundelkhand University Jhansi India; Dr. Rajesh Dabur , Department of Biochemistry  MDU, Rohtak, Amity Institute of Biotechnology, Amity University Rajasthan, India and finally  to all technical staff members for their wonderful lab support without any of them this work was not possible for me.

 

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Accepted on 18.05.2019          © RJPT All right reserved

Research J. Pharm. and Tech 2019; 12(9):4231-4234.