Standard Operating Procedures

The Standard Operating Procedures listed below are used in the Schistosomiasis Resource Center to maintain the life cycles of Schistosoma spp. The SOPs are divided into four categories.

Procedures common to maintaining intermediate snail hosts of Schistosoma spp

Artificial Pond Water for the cultivation of Biomphalaria, Bulinus and Oncomelania species
Aquaria maintenance
Growth of immature B. glabrata
Snail egg clutch collection
Growth conditions until patency
Control of rotifers and other commensals living in snail aquaria
Gel snail food preparation
Preparation of mud base medium for growth of Nostoc (cyanobacteria)
Shedding B.glabrata patent snails for cercariae

Procedures common to maintaining Schistosoma spp life cycles

Percutaneous exposure via the tail using Schistosoma spp cercariae
Perfusion of adult worms from infected mice
Behavior of Schistosoma mansoni cercariae
Preparation of in vitro schistosomules
Collection of lung Schistosoma mansoni schistosomules
Culturing schistosomula
Isolating Schistosoma spp. eggs from murine liver and gut
Microscopic evaluation of sporocyst development in live Biomphalaria glabrata
Shedding cercariae from patent snails
Exposure of snails to miracidia

Maintenance of S.japonicum in the Oncomelania hupensis spp. snail host

Artificial Pond Water for the cultivation of Biomphalaria, Bulinus and Oncomelania species
Care of breeding pairs of Oncomelania hupensis
Collecting, Isolating, and Hatching Oncomelania hupensis ssp. eggs
Shedding Oncomelania for S.japonicum cercariae

Maintenance of S.haematobium in the Bulinus truncatus spp. snail host

Artificial Pond Water for the cultivation of Biomphalaria, Bulinus and Oncomelania species
Maintenance of Bulinus truncatus truncatus snails
Exposure of hamsters to Schistosoma haematobium cercariae
Perfusion of S. haematobium from hamsters
Collection of S. haematobium eggs from hamsters

Molecular and cell culture products

Biomphalaria glabrata embryonic cell (Bge) cell culture
Genomic DNA extraction from schistosome intermediate hosts and parasite stages
Non-invasive DNA transfection to B. glabrata using polyethylenimine (PEI)
Preparation of soluble egg antigen (SEA) S.japonicum eggs
RNA extraction from schistosome intermediate hosts and parasite stages using RNAzol®

Other

Exposing Oncomelania snails to Schistosoma japonicum miracidia
Schistosoma japonicum cercarial collection and exposure of mice
Collecting Schistosoma japonicum eggs from mouse tissues
Growth of immature Biomphalaria glabrata
Preparation of water to house snails
Fixing Biomphalaria glabrata snails for dissection
Microscopic quantification of cercariae
A histochemical method for identifying sex of Schistosoma mansoni cercariae (under construction)
Percutaneous exposure (via the tail) of mice with Schistosoma cercariae
Collection of parasite eggs from tissues
Preparation of water to house Oncomelania hupensis
Molecular technique for sexing cercariae