Fumaria indica (L), a Famous Medicinal Herb of Tribal Regions of Jabalpur, Madhya Pradesh: Broad Spectrum Antibacterial and Phytochemical Profilng Against Some Pathogenic Microorganisms

Fumaria indica (L.) belongs to family Fumariaceae, and genus Fumaria commonly called shahtera “Pit papra” in the tribal vernacular. It is an annual herb and it’s distributed all over Asia, Europe and Africa. It is a familiar weed found in the plains of India. F. indica have been reported various medicinal purposes to acquire pharmacological activities like antipyretic1 hepatoprotective2 hypoglycemic3 antidiarrheal4 antispasmodic5 antihelmintic6 antieczema7 antiperiodic compound8 liver complaints9 and scrofulous skin affections.10 Infectious diseases have threatened the continued existence of humans since very early civilizations.11,12 The folk remedies, are still as an important part of traditional medicine13 presently many human diseases and infections are cured by a diversity of plants14 or plant derivative products.15 The main cause for the continuing attention and broad research on plants for antibacterial properties is the manifestation of challenging strains of bacteria.16,17 These strains are competent to survive with the same pace as their genetic evolution requires continuous development of new drugs against them.18 Therefore, bacteria in fastidious are imposing require for new drugs.19Infectious diseases are persistent and are major explanation of premature death all over world.20,21The prevalence of severe infections in human beings has significantly increased all over the world and it has become the leading cause of mortality in developing countries.22 About 80% population of the world relies on plants as a natural source of medicine.23 They are used medicinally in different countries and are a source of many potent and powerful drugs.24This study was aimed on validating the traditional use of selected medicinal plants against common bacteria, causing several human infections including Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, andKlebsiella pneumonia25-27by evaluating their in vitro antibacterial activity. The plants investigated in this study commonly used to treat the infectious diseases and the associated symptoms are listed in (Table 1).


INTRODUCTION
Fumaria indica (L.) belongs to family Fumariaceae, and genus Fumaria commonly called shahtera "Pit papra" in the tribal vernacular. It is an annual herb and it's distributed all over Asia, Europe and Africa. It is a familiar weed found in the plains of India. F. indica have been reported various medicinal purposes to acquire pharmacological activities like antipyretic 1 hepatoprotective 2 hypoglycemic 3 antidiarrheal 4 antispasmodic 5 antihelmintic 6 antieczema 7 antiperiodic compound 8 liver complaints 9 and scrofulous skin affections. 10 Infectious diseases have threatened the continued existence of humans since very early civilizations. 11,12 The folk remedies, are still as an important part of traditional medicine 13 presently many human diseases and infections are cured by a diversity of plants 14 or plant derivative products. 15 The main cause for the continuing attention and broad research on plants for antibacterial properties is the manifestation of challenging strains of bacteria. 16,17 These strains are competent to survive with the same pace as their genetic evolution requires continuous development of new drugs against them. 18 Therefore, bacteria in fastidious are imposing require for new drugs. 19 Infectious diseases are persistent and are major explanation of premature death all over world. 20,21 The prevalence of severe infections in human beings has significantly increased all over the world and it has become the leading cause of mortality in developing countries. 22 About 80% population of the world relies on plants as a natural source of medicine. 23 They are used medicinally in different countries and are a source of many potent and powerful drugs. 24 This study was aimed on validating the traditional use of selected medicinal plants against common bacteria, causing several human infections including Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, andKlebsiella pneumonia [25][26][27] by evaluating their in vitro antibacterial activity. The plants investigated in this study commonly used to treat the infectious diseases and the associated symptoms are listed in (Table 1).

Plant materials and extraction
The plant was collected from Bagrachi village of Jabalpur district. The collection was done in March, 2017. To confirm and authenticate the identified plant taxonomically, the samples were examined at State Forest Research institute (SFRI) under voucher no. 586. The plant was selected based on reports of its widespread use among the tribal communities. The collected plant materials were air-dried and finely powdered using a blender. To prepare methanol and chloroform extracts of the plant materials, 20 g of each powdered plant material was extracted with 200 ml of methanol and chloroform for 48 h at room temperature. The extracted suspensions were filtered through Whatman No. 1 filter paper (Himedia) and the filtrates were concentrated to dryness using a rotary evaporator, to yield the crude extract (Table.2) Test for Saponins (Foam test) 5 ml of crude extract was mixed with 20 ml of distilled water and then agitated in a graduated cylinder for 15 minutes. The formation of bubbles indicates the presence of saponins. 34 Test for Flavonoids(Alkaline reagent test) 2 ml of crude extract was treated with a few drops of 1N sodium hydroxide solution and observed the formation of strong yellow color. This yellow color becomes colorless on the addition of dilute hydrochloric acid, indicating the presence of flavonoids. 35

Test for Phenolic Compounds (Ferric chloride test)
Few drops of the extract were treated with 5% aqueous ferric chloride. The formation of deepblue or black color indicates the presence of phenolic compounds. 35 Test for Alkaloids (Mayer's Test) 2 ml of crude extract was treated with 2 drops of Mayer's reagent. The presence of white creamy precipitate indicates positive test. 35

Test for glycosides (Fehling's test)
An equal volume of Fehling A and Fehling B reagents were mixed together and 2ml of this solution was added to the crude extract and gently boiled. A brick-red precipitate appeared at the bottom of the test tube indicated the presence of reducing sugars. 35

Data analysis
The experiments were performed in triplicate and data were expressed as mean ± standard deviation (SD) .The data obtained was analysed using statistical package for social sciences (SPSS-Version 20).

RESULTS
The preliminary qualitative phytochemical screening of different extract showed maximum number of phytoconstituents along with alkaloids, terpenoids, steroids, flavonoids, phenols ( Table 3). The zone of inhibition of methanol and chloroform extracts of Fumaria indica on gram positive and negative bacteria at different concentrations, by using agar well diffusion method, was determined to access their antibacterial effect. Both extracts of Fumaria indica exhibited sensible antibacterial activity against five tested bacterial strains as compared to the standard antibiotic ciprofloxacin ( Table 4). The highest zones of growth inhibition were exhibited by methanol and chloroform extract against all the microorganisms compared to aqueous extract are shown in Institute of Microbial Technology, The bacterial procured cultures were sub-cultured in nutrient agar medium composition of (beef extract -3 g, NaCl -5 g, peptone -5 g, agar -15 g, distilled water -1000 ml) andincubated at 37°C. All the selected test strains were maintained and were sub-cultured after 20 days. The stock cultures of bacteria were maintained on nutrient agar slant at 4°C respectively.

Determination of antibacterial activity
Antibacterial activity was determined using well diffusion method. 29 Petri plates were prepared with 20 ml of sterile Muller Hinton agar media (HiMedia). Wells (6 mm diameter) were punched in the Muller Hinton agar and filled with plant extracts. The test cultures were swabbed on the top of the solidified media and allowed to dry for 10 min for compound diffusion. The tests were conducted at three different concentrations 50mg/ml, 100mg/ml and 150mg/ml of the crude extracts. Ciprofloxacin (25µl) was used as positive control. The plates were incubated for 18-24 hours at 37±1 0 C. Zone of inhibition was recorded in millimeters using transparent (HiMedia) antibiotic zone scale and the experiment was repeated by triplicates.

Phytochemical screening
The Preliminary qualitative phytochemical screening of Funmaria indica extracts was carried out by the standard methods. 30,31 Test for Steriods (LibermannBurchard Test) 1 ml of the crude extract was taken and dissolved in 10 ml of chloroform and an equal volume of concentrated sulphuric acid was added by sides of the test tube. The upper layer turns red and the sulphuric acid layer showed yellow with green fluorescence. These indicate the presence of steroids. 32 Test for Terpenoids (Salkowski test) 2 ml of crude extract was taken and added to 2 ml of acetic anhydride and absorption of H 2 SO 4 . Formations of blue, green rings indicate the presence of terpenoids. 33 Dry weight of plant material Dry weight of extract Percentage Yield (%) =     of growth inhibition was observed on both extracts against Bacillus subtilis which gave a zone of inhibition measuring 1.33±2.30 and 0.00± 0.00 respectively at a dose of 50 mg/ml shown in (Table 4). The aqueous extract does not show any zone of inhibition.

DISCUSSION
Several studieshave been confirmed for the extracts of plant species possessed activity with regard to antimicrobial properties 36

CONCLUSION
The results of the present study reveal that the antibacterial activity of traditional medicinal herb fumaria indica extracts against all the selected pathogenic bacterial strains is remarkably fascinating. This herb thus justifies its use by tribal to treat various diseases like digestion, diarrohea, fever, inflammations, liver complaints, skin infection and vomiting. This success in the beneficial role of native tribal communities hence strongly stems the fact that this plant bears the capability of possessing most important bio-conjugated constituents that could serve as a source of novel drug design formulation.

ACKNOWLEDGEMENT
Author is grateful to the University Grant Commission, New Delhi for providing (RGNF) Rajeev Gandhi National Fellowship for this study.
We are thankful to the Head, Department of Post-Graduate Studies and Research in Biological Science, Rani Durgavati University, Jabalpur for providing laboratory facilities.

CONFLICTS OF INTEREST
No conflicts of interest.