caa a22 4500 445343508 CHVBK 20180317142821.0 cr unu---uuuuu 170323e20110201xx s 000 0 eng 10.1007/s00436-010-2083-8 doi (NATIONALLICENCE)springer-10.1007/s00436-010-2083-8 Comparison of protein-free defined media, and effect of l -cysteine and ascorbic acid supplementation on viability of axenic Entamoeba histolytica [Elektronische Daten] [Weng Wong, Zi Tan, Boon Lim, Zeehaida Mohamed, Alfonso Olivos-Garcia, Rahmah Noordin] Entamoeba histolytica is the etiologic agent for amoebiasis. The excretory-secretory (ES) products of the trophozoites contain virulence factors and antigens useful for diagnostic applications. Contaminants from serum supplements and dead trophozoites impede analysis of ES. Therefore, a protein-free medium that can sustain maximum viability of E. histolytica trophozoites for the longest time duration will enable collection of contaminant-free and higher yield of ES products. In the present study, we compared the efficacy of four types of media in maintaining ≥95% trophozoite viability namely Roswell Memorial Park Institute (RPMI-1640), Dulbecco's Modified Eagle Medium (DMEM), phosphate-buffered saline for amoeba (PBS-A), and Hank's balanced salt solution (HBSS). Concurrently, the effect of adding l-cysteine and ascorbic acid (C&A) to each medium on the parasite viability was also compared. DMEM and RPMI 1640 showed higher viabilities as compared to PBS-A and HBSS. Only RPMI 1640 showed no statistical difference with the control medium for the first 4h, however the ≥95% viability was only maintained for the first 2h. The other protein-free media showed differences from the serum- and vitamin-free TYI-S-33 control media even after 1h of incubation. When supplemented with C&A, all media were found to sustain higher trophozoite viabilities than those without the supplements. HBSS-C&A, DMEM-C&A, and RPMI 1640-C&A demonstrated no difference (P > 0.05) in parasite viabilities when compared with the control medium throughout the 8-h incubation period. DMEM-C&A showed an eightfold increment in time duration of sustaining ≥95% parasite viability, i.e. 8h, as compared to DMEM alone. Both RPMI 1640-C&A and HBSS-C&A revealed fourfold and threefold increments (i.e., 8 and 6h, respectively), whereas PBS-A-C&A showed only onefold improvement (i.e., 2h) as compared to the respective media without C&A. Thus, C&A-supplemented DMEM or RPMI are recommended for collection of ES products. Springer-Verlag, 2010 Wong Weng Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, 16150, Kubang Kerian, Kelantan, Malaysia aut Tan Zi School of Health Sciences, Universiti Sains Malaysia, 16150, Kubang Kerian, Kelantan, Malaysia aut Lim Boon School of Health Sciences, Universiti Sains Malaysia, 16150, Kubang Kerian, Kelantan, Malaysia aut Mohamed Zeehaida Department of Medical Microbiology and Parasitology, School of Medical Sciences, Universiti Sains Malaysia, 16150, Kubang Kerian, Kelantan, Malaysia aut Olivos-Garcia Alfonso Departamento de Medicina Experimental, Facultad de Medicina, Universidad Nacional Autónoma de México, 04510, México, Mexico DF, Mexico aut Noordin Rahmah Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, 11800, Penang, Malaysia aut Parasitology Research Springer-Verlag 108/2(2011-02-01), 425-430 0932-0113 108:2<425 2011 108 436 https://doi.org/10.1007/s00436-010-2083-8 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00436-010-2083-8 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Wong Weng Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, 16150, Kubang Kerian, Kelantan, Malaysia aut NATIONALLICENCE 700 1- Tan Zi School of Health Sciences, Universiti Sains Malaysia, 16150, Kubang Kerian, Kelantan, Malaysia aut NATIONALLICENCE 700 1- Lim Boon School of Health Sciences, Universiti Sains Malaysia, 16150, Kubang Kerian, Kelantan, Malaysia aut NATIONALLICENCE 700 1- Mohamed Zeehaida Department of Medical Microbiology and Parasitology, School of Medical Sciences, Universiti Sains Malaysia, 16150, Kubang Kerian, Kelantan, Malaysia aut NATIONALLICENCE 700 1- Olivos-Garcia Alfonso Departamento de Medicina Experimental, Facultad de Medicina, Universidad Nacional Autónoma de México, 04510, México, Mexico DF, Mexico aut NATIONALLICENCE 700 1- Noordin Rahmah Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, 11800, Penang, Malaysia aut NATIONALLICENCE 773 0- Parasitology Research Springer-Verlag 108/2(2011-02-01), 425-430 0932-0113 108:2<425 2011 108 436 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer