Background: Permanent staining of helminths is important for different purposes including taxonomic
and prevalence studies as well as teaching. Carmine is one of the most commonly used dyes; however,
variable factors affect the quality of the staining outcomes. Therefore, it is essential to find an alternative
stain that is efficient, accurate, and less complex.
Objective: The aim of the present study is to evaluate the quality of a combined lactophenol methylene
blue-blue glycerol jelly (LMB-BGJ) technique as a potential alternative for staining of helminths compared
with carmine-based staining and glycerol jelly mounting techniques.
Material and Methods: S. mansoni and T. spiralis were obtained from experimentally infected animals. A
new staining technique (LMB-BGJ) was used for both parasites in comparison with carmine stain in case of
S. mansoni and glycerol jelly mounting in case of T. spiralis. Comparison was conducted using examination
under light microscope and a mean score was given for the structural brightness, and extent of staining of
tegument/cuticle, suckers, intestine, as well as reproductive systems.
Results: S. mansoni worms prepared with both carmine staining and LMB-BGJ showed moderate staining
quality of tegument, suckers, intestine and reproductive organs with a mean total score of 3.3/4 and
3.35/4, respectively. Adult T. spiralis mounted with glycerol jelly only showed a mean total score of 3.95/4
for the cuticle, esophagus, and intestine, while the LMB-BGJ showed a staining quality with a mean total
score of 3.85/4.
Conclusion: From a practical point of view, LMB-BGJ is a simple and rapid technique that needs less
experience for the preparation of semi-permanent slides with satisfactory staining and mounting quality
for both trematodes and small nematodes.