Assessment of the viability of Schistosoma mansoni schistosomula by comparative uptake of various vital dyes

Gold, Daniel

doi:10.1007/s004360050227

Cited by 27 publications

(19 citation statements)

References 14 publications

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“…In addition, according to the literature, using the propidium iodide staining methods to differentiate dead and viable eggs has been evidenced by such limitations in the viability tests for the other parasites such as Trichuris muris eggs (failure to identify nonviable eggs, Kopper and Mansfield 2010) and Cryptosporidium andersoni oocysts (dependence to the storage medium, Kváč et al 2007). Also, Gold (1997) compared the uptake of two vital dyes, toluidine blue and methylene blue, for the assessment of the viability of Schistosoma mansoni schistosomula. That study proved that the main disadvantage of toluidine blue was rapid, nonspecific staining of live schistosomula, whereas methylene blue did not always stain flattened, dead schistosomula or it stained them an uncontrasting pale blue.…”

Section: Discussionmentioning

confidence: 99%

In vitro viability test for the eggs of Echinococcus granulosus: a rapid method

Moazeni

Rakhshandehroo

2011

Parasitol Res

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In this study an attempt was made to develop an efficient, rapid, simple, and reproducible method for the in vitro viability test of Echinococcus granulosus eggs. The eggs were obtained from an experimentally infected dog and kept at 4°C until use. To prepare the dead or damaged eggs, the eggs were heated in hot water (69-72°C for 10 min), preserved in 70% ethyl alcohol (16 days) or exposed to direct sunlight (18 h). Sodium hypochlorite (0.5-0.7%) was used for the hatching process, and the hatched oncospheres were stained with 0.1% eosin for the viability test. With 0.5% sodium hypochlorite, the hatching rates for viable eggs and eggs killed or damaged by heat (69°C), 70% ethyl alcohol, and direct sunlight were 96%, 97.5%, 91.5%, and 94.6% respectively and there was no significant difference between the hatching rate for viable and dead or damaged eggs (p> 0.05). After staining with 0.1% eosin, the rates of the viable oncospheres hatched from viable eggs and the eggs killed or damaged by heat (69°C), 70% ethyl alcohol, and direct sunlight were 97.5% 3.6%, 7%, and 10.5%, respectively. The difference between the rates of viable oncospheres hatched from viable and dead or damaged eggs was extremely significant (P < 0.0001). With 0.7% sodium hypochlorite, the hatching rates for viable and dead eggs (killed by 72°C for 10 min) were 99.1% and 99.9%, respectively. In this condition, the rate of viable oncospheres was an average of 98.5% for viable eggs and 0.0% for dead ones. The results of this study showed that hatching of eggs by 0.7% sodium hypochlorite and staining of hatched oncospheres by 0.1% eosin are practical methods for the differentiation of viable and nonviable (dead) eggs of Echinococcus granulosus.

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Section: Discussionmentioning

confidence: 99%

In vitro viability test for the eggs of Echinococcus granulosus: a rapid method

Moazeni

Rakhshandehroo

2011

Parasitol Res

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“…As shown in Table 1, many of the knocked down genes presented the general phenotype of reduced motility, while the activity of enzymes, demonstrated by biochemical assays, were also reduced. Further characterization of the tegument integrity could have involved the exclusion of vital dyes [43] or ultrastructural analysis, which would have provided information on a complex and important region of the parasite [44][45][46]. This latter approach has been used to demonstrate the involvement of tetraspanins in maintaining tegument fidelity [47].…”

Section: Rnai As a Tool To Investigate Schistosome Biologymentioning

confidence: 98%

Gaining biological perspectives from schistosome genomes

Gobert

You

McManus

2014

Molecular and Biochemical Parasitology

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“…Over the past many years, the conventional Trypan Blue has been used for assessing cell viability of various parasites and cells as well (Gold 1997;Wiehart et al 2006;Jin et al 2009;Tan et al 2010). MF has been used widely on malaria (Field 1940;Fenton and Innes 1945;Reilly et al 1997), filaria (Sivanandam and Mak 1975), Leishmania (Chunge et al 1989), Acanthamoeba spp.…”

Section: Discussionmentioning

confidence: 99%

Modified Field stain - rapid viability test for Trichomonas vaginalis

Afzan

Sivanandam

2011

Journal of Applied Microbiology

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Aims: We previously reported that Modified Field Stain (MF) can be used as a rapid stain for diagnosis. In the present study we extend the observation to include the stain as an alternative method to assess viability of the cells. Methods and Results: Six isolates of Trichomonas vaginalis were used to assess the utility of the Modified Field stain as a rapid viability test for T. vaginalis and to compare with 0AE4% Trypan Blue dye exclusion test in three conditions; normal in vitro culture growth using Hollander medium, lysed in distilled water and treated with metronidazole. MF stain showed similar growth profile pattern as Trypan Blue dye exclusion for identifying viable cells of T. vaginalis. Although, Trypan Blue dye exclusion test is ready made, rapid and widely used in laboratory as reliable viability assay, however, the limitation using Trypan Blue is the dye was unable to show internal morphological changes during the parasite's transition from being viable to non-viable. On day 3 where cultures peaked the correlation factor of both assays done to assess the viability of parasites harvested from the controls, metronidazole and distilled water treated parasites were more than 0AE9 respectively. Conclusions: This confirms that MF staining does not only record permanently the morphological changes and retain internal structural details but also provides a reliable and rapid viability assay for the parasites. Significance and Impact of the Study: Therefore, in our study, Modified Field's stain may offer the researchers and laboratory technologists the opportunity to get the result on the same day and the most important thing is the ability to differentiate between viable and non-viable of T. vaginalis under three different conditions (normal culture, drug and distilled water condition). Modified Field's staining method enhanced the morphological identification of T. vaginalis compared to Trypan Blue dye exclusion.

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Assessment of the viability of Schistosoma mansoni schistosomula by comparative uptake of various vital dyes

Cited by 27 publications

References 14 publications

In vitro viability test for the eggs of Echinococcus granulosus: a rapid method

In vitro viability test for the eggs of Echinococcus granulosus: a rapid method

Gaining biological perspectives from schistosome genomes

Modified Field stain - rapid viability test for Trichomonas vaginalis

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