Aim:An environment compatible technique to stain Platyhelminthes, Fasciola gigantica, Gastrothylax crumenifer, Taenia solium, and Moniezia expansa using aqueous and alcoholic extract of sugar beet (Beta vulgaris), China rose (Hibiscus rosa-sinensis), and red rose (Rosa hybrida) were described to minimized the deleterious effects of the synthetic dyes.Materials and Methods:Aqueous/ethanolic extracts of roses were extracted from the flowers while red beet was extracted from the roots.Results:Stained helminthes acquired a comparable level of pigmentation with the distinction of their internal structure in these natural dyes. The flukes (liver and rumen) internal structure, oral and ventral/posterior sucker, cirrus sac, gravid uterus, testes, ovary, and vitallaria were appeared pink color in aqueous and alcoholic extract of either China or red rose and yellow to brown color in sugar beet stain. The interior of the proglottid of T. solium and M. expansa took yellow to brown color with good contrast in sugar beet stain and of pink to pink-red in China and red rose stain.Conclusion:The extract of roses (red rose followed by China rose) followed by red beet possess the potential to replace the conventional stains in the taxonomic study of Platyhelminthes parasites.
Aim:Preservation of macroparasites by infiltrating the polymer in the tissues can defy the inherited shortcoming of classical wet preservation method.Materials and Methods:Preservation was done by infiltrating the melamine alone or with xylene (MX)/chloroform (MC)/turpentine oil (MT) in 1:1 and hardener (MH) in 9:1 ratio in the tissues of the gross specimen of the animal parasites.Results:The plastinated models withstand the process of microbial decomposition, and remain intact in the environmental conditions. The polymer mixture resists the entry of the water molecule, and model dried just after taking out it from the water tank. Overall, the plastinated parasites were dry, non-sticky, glossy, odorless, chemical free, and harmless, to some extent flexible, with detectable morphological structure, and retain their natural form but lost their natural color. Full marks were assigned to the degree of dryness, non-stickiness, and odorlessness to the model plastinated in different solutions on a five-point scale. For flexibility, the score was 1.2, 2.2, and 2.4 for the plastinated model in melamine/MH, MX/MC, and MT solutions, respectively. The average score of glossiness was 4.6 and 5 for the specimen plastinated in melamine/MH and MX/MC/MT solutions, respectively. The degree of dryness, glossiness, stickiness, and flexibility varies non-significantly, with the polymer mixtures used.Conclusion:The prepared model can be used to educate the students/general mass population.
The objective of this study was to characterize Haemonchus contortus antigens, and to standardize and evaluate indirect plate and dot-ELISA using homologous antigens in the small ruminants. Electrophoretic separation of somatic antigen in reducing condition on 15% polyacrylamide gel resolved into 16 proteins of molecular weight ranging from 14-100 kDa. Two step ethanolic extraction of the supernatant of in-vitro culture of H. contortus yielded excretory-secretory (E-S) antigen/ cathepsin L cysteine proteinase of molecular weight 28 kDa. The animals (Goats=103; Sheep=91) were broadly kept into post-mortem (PM) and faecal examined groups and further sub-grouped based on mono or multiply helminths infection. At many occasion, the somatic antigen found to cross reacts with other helminths parasites thus minimizing the specificity of the selected tests and antigens. There was no any direct correlation between the parasites load and ELISA reactivity pattern. The prevalence rate of haemonchosis was 55.7 (34/61) in goats/ 47.6 (40/84) % in sheep as per PM examination while it was 45.63 (47/103) in goats/ 41.76% (38/91) in sheep and 36.89 (38/103) in goats/ 35.16% (32/91) in sheep using E-S antigen based plate and dot-ELISA, respectively. With E-S antigen, the overall % sensitivity, specificity, positive and negative predictive values of plate-ELISA was 89.74 (goats)/ 80.95 (sheep), 81.25 (goats)/ 91.84 (sheep), 74.47 (goats)/ 89.47 (sheep), 92.86 (goats)/ 84.91 (sheep), respectively while for dot-ELISA it was 66.67 (goats)/ 61.9 (sheep), 81.25 (goats)/ 87.76 (sheep), 68.42 (goats)/ 81.25 (sheep), 80 (goats)/ 72.88 (sheep), respectively, so the tests and E-S antigen can be recommended for the detection haemonchosis in the small ruminants.
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