Appendix 1. Steps followed during preparation of permanent slides (Slides a – m)

Slide a. Collected parasites in petridish

Slide b. Relaxation of collected parasites in physiological saline for 1 hr

Slide c. Fixation of parasites in 10% formalin

Slide d . Washing of parasites in running water for 6 hrs

Slide e. Staining the parasites with Borax Carmine for overnight

Slide f. De-stain the parasites in acid alcohol until the stain removed

Slide g. Dehydration in 90% ethanol for 15 minutes

Slide h. Dehydration in 95% ethanol for 15 minutes

Slide i. Dehydration in absolute alcohol for 30 minutes

Slide j. Dehydration in acid alcohol for 30 minutes

Slide k. Cleared in Lectophenol for 15 minutes

Slide l. Mounted in Canada Balsam Slide m. Prepared permanent slides