Challenges Involve in Metacercariae Extraction, Isolation and Preservation for Experimental Research.
Dauda Iliyasu, Rwuaan J Sankay, Chiezey N Paulin, Ate I Utem, Okubanjo O Olayinka, Ahamed Ishiyaku Musa, Ajani J Ayinla.
Abstract The study was designed to evaluate the challenges involve in extraction, isolation and preservation of metacercariae for research purposes. Cercariae is one of the active larval stage in the life cycle of Fasciola. Metacercariae is the encysted cercariae which served as intermediate host, commonly found in snails Lymnea natalensis and Lymnea auricularia in the tropics. One hundred (100) naturally infested snails of Lymnea natalensis and Lymnea auricularia was collected from river barks and streams for extraction of metacercariae. The snails were crushed in a petri dish using forceps and a dissection pin. The shells were removed while the tissues were washed with water and sieved. The filtrate was left uncovered in a Petridish for 72 hours, and the supernatant discarded. The petri dishes were emptied of water and kept at 4 0C in a refrigerator. The attached metacercariae on the petri dish were sprinkle with water every 48 hours to sustain their viability. The attached metacercariae on the petri dish were scraped into 40 ml of distilled water in a cylinder and mixed, 0.1 ml of the mixture at 3 slide reading revealed the percentage of viable and dead metacercariae 33.33 % 38.14 % and 28. 52 % and 35.59 %, 25.42 % and 38.98 % respectively. It was concluded that moisture and refrigerator temperature 4 0C are key for isolation and preservation of viable metacercariae.
Key words: Lymnea natalensis, Lymnea auricularia, Cercariae, Extraction of metacercariae, Isolation, Preservation and viability.