Dear Ashish,

Herein some of the anthelmintic activity screening methods:

1- Evaluation of Anthelmintic Activity with Housefly Worms

In each of the 10 Petri dishes containing 20 mL of the prepared, solidified, and sterilized housefly medium, a well having 1 cm diameter was made with the help of a borer at the centre. The freshly collected matured female houseflies were placed in each of the Petri dishes and covered. After 24 hours, eggs hatched out from the worms, which were white in color, and died houseflies were removed. The Petri dishes were cultured further for a period of 72 hours, resulting in the formation of worms with an average count of 32 worms in each Petri dish. From these only 25 worms from each Petri dish were taken for experiments and the remaining worms were removed from all Petri dishes.

The cultured Petri dishes were named for standard drug (SD) and test drugs (TD1 and TD2) with control. The concentrations of the drugs were noted on the respective Petri dishes. Drug solution (0.2 mL) was added to each and every well in each Petri dish, and the time was noted for standard and test drugs with control. Albendazole solution was used as reference standard drug and distilled water as control. All the Petri dishes were incubated at 37°C. The worms were observed for motility. This was done after tapping the edges of the Petri dishes and allowing the worms to move freely towards the well, the worms that were alive would be seen moving. After paralysis, the arrest in the movement of the worms was clearly observed, while applying little pressure with tapped pen edges. The unparalysed motile worms were trapped towards the well, and the incubation process was carried out again. In the untreated control Petri dishes worms were viable for at least 10 days. The number, weight, age of worms, motility, paralytic activity, and death time were noted.

Collection of Earthworms

Earthworms (Pheretima posthuma) were collected from the swampy water along the Muthukur road, near Jagan’s College of Pharmacy, Nellore, and Andhra Pradesh, India. The average size of worms was 5–8 cm. Earthworms were identified and authenticated by Dr. Humanan Reddy, Professor of Entomology, Department of Zoology, Vikrama Simhapuri University, Nellore. The anthelmintic activity was carried out as per the method described elsewhere [9]. The assay was performed in vitro using adult earthworms owing to their anatomical and physiological resemblance with the intestinal round worms, parasites of human beings for preliminary evaluation of anthelmintic activity. A concentration of standard drug (albendazole) and synthetic test drugs (APMT and APTT) was prepared as described earlier.

Evaluation of Anthelmintic Activity Using Earthworms

Earthworms, each of average length of 6 cm, were placed in Petri dishes containing 2 mL of various drug concentrations, 12.5 mg/mL, 25 mg/mL, 50 mg/mL, 100 mg/mL, and 200 mg/mL, of solutions. Albendazole solution was used as reference standard drug and distilled water as control. The worms were observed for the motility after incubating at 37°C. This was done after pouring the Petri dishes content in the wash basin and allowing the worms to move freely. By tapping the end of each worm with the index finger and applying a bit of pressure, the worms that were alive showed motility and those dead were nonmotile. The motile worms were returned to the respective Petri dishes containing drug solutions, and the incubation process was carried out again. In the control, the worms were viable for at least twelve days, which is similar to the findings reported earlier [10–12]. The time taken for paralysis, motility activity of any sort, and death time of worms were observed and recorded after ascertaining that the worms did not move neither when shaken vigorously nor when dipped in warm water (50°C).

http://www.hindawi.com/journals/isrn/2012/709860/

2-The anthelmintic activity was performed according to the method Ghosh et al. on adult Indian earth worm Pheretima posthuma as it has anatomical and physiological resemblance with the intestinal roundworm parasites of human beings. Pheretima posthuma was placed in petridish containing four different concentrations (25, 50, 75 and 100mg) each of A. sessilis (methanol, acetone and water) extract solutions. Each petridish was placed with 3 worms and observed for paralysis and death. The mean time for paralysis was noted when no movement

of any sort could be observed, except when the worm as shaken vigorously; the time death of worm (min) was recorded after ascertaining that worms neither moved when shaken nor when given external stimuli. In the same manner albendazole was included as reference compound. The test results were compared with reference compound albendazole (15mg/ml) treated samples. 

Ghosh T, Maity TK, Bos A, Dash GK. Anthelmintic activity of Bacopa monierri. Ind

Journal of Natural Products, 2005; 21(2): 16-19.

3- Evaluation of anthelmintic activity: The anthelminthic experiment was carried as per the method described by Mali et al. (2005) with minor modifications using adult earthworm (Pheretima posthuma) owing to its anatomical and physiological resemblance with the intestinal roundworm parasites of human beings for preliminary evaluation anthelmintic activity. Test sample of the extract of different concentrations (15, 30 and 45 mg mL–1) were prepared in ethanol and 7-8 worms of Pheretima posthuma of 8-10 cm were placed petri dish containing 25 mL of above test solution of extract (Kosalge and Fursule, 2009). Albendazole (15 mg mL–1) was used as reference standard and normal saline (0.9% NaCl) as control and 25 mL of ethanol was also taken for consideration as all the extracts were prepared in same solvent and the effect of it should be analyzed. All the test solution and standard drug solution were prepared freshly before starting the experiment. Observation were made for the time taken for paralysis noted when no movement or loss of movement. Time for death of worms were recorded after controlling that worms neither moved when shaken vigorously nor when dipped in warms water and fading away the color of warms. All the result were shown in table and expressed as a mean of selected worms in each group.

Statistical analysis: The calculation of the experimental data is done by using One-Way analysis of variance (ANOVA). Later on the calculation is done by various non parametric test. The mean and standard deviation is obtained. The results were presented as Mean±SEM. Differences between means of treatment and control groups were accepted significant at p

rafik karaman sir, i wanted to ask: why did u suggest the housefly worms to be used for anthelmintic activity? how does it resembles true worms that infect the body of a human being or an animal? i'm interested to use the method but i don't know how to correlate it with the true worms. aren't these housefly worms commonly known as the "maggots"?