A Quick Look At Anticonvulsant Study Biology Essay

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Epilepsy is the collective term used for a group of chronic seizure disorders which is having in common, sudden and transient episodes (seizure of loss or disturbance of consciousness), usually but always with a characteristic body movement (convulsion) and sometimes with autonomic hyper activity. The seizure nearly always correlates with an abnormal electrical discharge.

Drugs which antagonised convulsion induced by MES, Leptazol and Kindling Method are usually useful in Grand-Mal.

The experimental protocol (MES Method in Rats)94

In the present study anti-convulsant activity were screened by MES method. Albino rats of either sex were selected by random sampling technique was used for study. The animals were divided in three groups containing 4-5 animals in each group. One group was used as control, second as standard drug (phenytoin) and third was test group (synthesized compounds)

Different stages of the convulsions were noted as (a) tonic flexion, (b) tonic extensor phase, (c) clonic convulsions, (d) stupor, and (e) recovery or death. The animal was held properly and corneal electrodes were placed on the cornea and the prescribed current(150 mA for 0.2 seconds) was applied after half an hour administration of the test compounds. Phenytoin at the dose of 25 mg/kg i.p. was administered as standard drug for comparison. The test compounds at two dose levels were administered orally. The time spent by animal in each phase of convulsion was recorded. The reduction in time and abolition of tonic extensor phase was recorded. Animals in which extensor phase was abolished were taken as protected animals. The percentage protection is calculated as follows,

% protection = [(control- test)/control] - 100


6.2.1 In-vitro antibacterial study

The microbial world comprises of various micro-organisms which are microscopic in size. Bacteria, fungi (yeast and moulds) and microscopic algae are some of microorganisms. These are distinguished into two broad groups such as prokaryotes and eukaryotes. Eukaryotes contain nucleus and organelles (such as chloroplast, lysosome, endoplasmic reticulum, mitochondrion and golgi bodies) whereas, prokaryotes lacks the above features.

Bacteria are most abundant prokaryotic organism that is vital to life of living things. In nature bacteria can adapt to any kind of living conditions than any other group of organisms.

The following conditions must be accomplished for the determination of proper antimicrobial activity:

There should be proper control between the test organism and the substance to be evaluated.

The required conditions for the growth of the microorganism should be provided. Measurement of activity should be done correctly.

Aseptic should be maintained.

Study conditions should be maintained unchanged throughout the experiment.

Materials and methods

Various methods have been used to evaluate the antimicrobial activity of the drugs. The activity can be evaluated by the following techniques.

Agar streak dilution method.

Serial dilution method.

Agar diffusion method.

Cup plate method

Cylinder method

Paper disc method.

Turbidimetric method.


The standard strains were procured from the American type culture collection (ATCC), Rockville, USA, and the pathological strains were procured from the department of microbiology, CEEAL analytical lab, Chennai, India. The anti-microbial activity of the synthesized compounds was screened against the following bacteria.

Gram-positive organism:

Staphylococcus aureus (ATCC 6538P)

Bacillus substillis (ATCC 6633)

Gram negative organism:

Escherichia coli (ATCC 25922)

Pseudomonas aeruginosa (ATCC 25619)


Nutrient agar medium (hi-media laboratories, India) is used as the media for the study of anti-bacterial activity. The composition of the medium is as follows.



Peptic digest of animal tissue


Beef extract


Sodium chloride




Yiest extract




The anti-bacterial activity of the compounds Ia-Va and Ib-Vb were studied by the paper disc diffusion method. The test compounds were used in the concentration of 25µg/ml, 50µg/ml, 100µg/ml.

Ampicillin 50µg/ml was used as standard.

Disc diffusion method96

A suspension of Staphylococcus aureus was added to the sterile nutrient agar at 45°C in aseptic environment, the mixture was transferred to sterile petridishes and allowed to solidify. Sterile discs of Whatmann filter paper 6mm in diameter was dipped in solutions of compounds Ia-Va and Ib-Vb, standard were placed on the surface of the agar plates.

All the plates were allowed to stand at room temperature for 1 hour, (This was as a period of preincubation diffusion to minimize the effects of variation in time between the application of the different solutions) then plates were placed for incubation for 18 hours at 37 ± 1°C and observed for the antibacterial activity. In which plate zone of the inhibition observed diameter of that was measured.

Similar procedures was carried out for studying the antibacterial activity of the compounds Ia-Va and Ib-Vb against. The average area of the zone of inhibition was calculated and compared with the standard.

6.2.2 In-vitro Antifungal study

A fungus is a colorless plant which is lack of chlorophyll. Fungi that cause infections may be like yeast or hyphi (mould) and these are called fungal infections.

These are of two types

Superfacial mycotic infection

Systemic mycotic infection

Because of the large widespread prevalence and airborne and soilborne transmission of the fungal pathogens, sanitary methods are not sufficient to eradicate the disease caused. So the search for the new and improved agents continues.

Material and Method

By Disc Diffusion Method the antifungal activity of the synthesized compounds Ia-Va and Ib-Vb were studied. For this following organisms were used.

Aspergillus fumigates (ATCC 46645)

Candida albicans (ATCC 10231)

Compounds Ia-Va and Ib-Vb were used in the concentration of 25µg/ml, 50µg/ml, 100µg/ml using DMF as solvent.

Ketoconazole (50µg/ml) was used as standard.

The Disc Diffusion Method was employed for the screening of antifungal activity by using Sabouraud Dextrose Agar medium (30gms of sabouraud dextrose agar and 1000ml of water heating).

Disc Diffusion Method

A suspension of micro-organism (Aspergillus fumigates) was added to sterile Sabouraud dextrose agar medium at 45°C and the mixture was transferred to sterile petridishes and allowed to solidify. Sterile disc of Whatmann filter paper of 6mm in diameter dipped in solutions of synthesized compound Ia-Va, Ib-Vb and standard were placed on the surface of agar plates.

All the plates were allowed to stand at room temperature for 1hour. Then the plates were incubated at 37 ± 1°C for 18 hours and observed for antifungal activity. The diameter of zone of inhibition was measured. Similar procedure was carried out for studying antifungal activity of compounds against Candida albicans. The average area of zone of inhibition was calculated and compared with that standard.

Determination of Minimum Inhibitory Concentration

The MIC (minimum inhibitory concentration) of the synthesized compounds Ia-Va, Ib-Vb were determined by the agar streak dilution method. The MIC was determined against the bacteria and fungi.


Gram +ve

Staphylococcus aureus (ATCC 6538P)

Bacillus Substilis (ATCC 6633)

Gram -ve

Escherichia coli (ATCC 25922)

Pseudomonas aurogenousa (ATCC 25619)


Aspergillus fumigates (ATCC 46645)

Candida albicans (ATCC 10231 )

Media used

Media used for bacteria was nutrient agar and for fungi saboraud dextrose agar. All the culture media were sterilized by autoclaving at 15lbs for 20 min.

Agar Streak Dilution:97

The stock solutions (1mg/ml) of the synthesized compounds were made by using DMF as solvent. From this stock solution, required quantities of drug solution were mixed with the known quantities of the molten sterile agar media aseptically to provide the following concentration 1,2,3,4,5,6,7,8,9,10,15,20,25,30,35,40,45,50 and 100µg/ml. About 20ml of the media containing the drug was dispensed into sterile petridishes. Then the media were getting allowed to solidify.

Over the surface of the agar plates, 1µl of standardized micro-organism (1-105 CFU/ml) were poured aseptically. After inoculation, all the plates were incubated at 37 ± 10C for 24 hours. Then all the plates were observed for the growth of the organism. The lowest concentration of the synthesized compounds inhibiting the growth of the bacteria/ fungi were taken as the minimum inhibitory concentration of the test compounds against that bacteria/ fungi.