Media composition to revive leptospires stored at −80 °C
Leptospires are preserved by frequent sub-culturing in semisolid media due to the challenge of low recovery by freezing or liquid nitrogen methods. The present study evaluated three liquid EMJH medium compositions (Medium A: Leptospira medium base EMJH, Leptospira enrichment EMJH, 5-fluorouracil (3%...
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2022
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my.upm.eprints.1021622023-07-12T01:18:20Z http://psasir.upm.edu.my/id/eprint/102162/ Media composition to revive leptospires stored at −80 °C Philip, Noraini Neela, Vasantha Kumari Leptospires are preserved by frequent sub-culturing in semisolid media due to the challenge of low recovery by freezing or liquid nitrogen methods. The present study evaluated three liquid EMJH medium compositions (Medium A: Leptospira medium base EMJH, Leptospira enrichment EMJH, 5-fluorouracil (3%), rabbit serum (1%) and calf serum (1%); Medium B: same as Medium A but without 5-fluorouracil; Medium C: same as Medium B but with the addition of sodium pyruvate) for the revival of leptospires after storage at −80 °C. A total of 18 Leptospira serovars cultured in Medium A was aliquoted into cryogenic vials and directly stored at −80 °C. A hundred microlitre from each serovar culture stored at −80 °C was sub-cultured on a selected time over a period of 30 months into Media A, B and C. Regrowth on Media B and C showed a better and faster recovery (89–100%) (p-value <0.05) compared to Medium A (67–100%). Leptospires can be stored longer at −80 °C and a good recovery could be obtained when sub-cultured on EMJH medium without 5-fluorouracil. Academic Press 2022 Article PeerReviewed Philip, Noraini and Neela, Vasantha Kumari (2022) Media composition to revive leptospires stored at −80 °C. Cryobiology, 109. 89 - 93. ISSN 0011-2240; ESSN: 1090-2392 https://www.sciencedirect.com/science/article/pii/S0011224022000992?via%3Dihub 10.1016/j.cryobiol.2022.09.007 |
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Leptospires are preserved by frequent sub-culturing in semisolid media due to the challenge of low recovery by freezing or liquid nitrogen methods. The present study evaluated three liquid EMJH medium compositions (Medium A: Leptospira medium base EMJH, Leptospira enrichment EMJH, 5-fluorouracil (3%), rabbit serum (1%) and calf serum (1%); Medium B: same as Medium A but without 5-fluorouracil; Medium C: same as Medium B but with the addition of sodium pyruvate) for the revival of leptospires after storage at −80 °C. A total of 18 Leptospira serovars cultured in Medium A was aliquoted into cryogenic vials and directly stored at −80 °C. A hundred microlitre from each serovar culture stored at −80 °C was sub-cultured on a selected time over a period of 30 months into Media A, B and C. Regrowth on Media B and C showed a better and faster recovery (89–100%) (p-value <0.05) compared to Medium A (67–100%). Leptospires can be stored longer at −80 °C and a good recovery could be obtained when sub-cultured on EMJH medium without 5-fluorouracil. |
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Article |
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Philip, Noraini Neela, Vasantha Kumari |
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Philip, Noraini Neela, Vasantha Kumari Media composition to revive leptospires stored at −80 °C |
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Philip, Noraini Neela, Vasantha Kumari |
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Philip, Noraini |
title |
Media composition to revive leptospires stored at −80 °C |
title_short |
Media composition to revive leptospires stored at −80 °C |
title_full |
Media composition to revive leptospires stored at −80 °C |
title_fullStr |
Media composition to revive leptospires stored at −80 °C |
title_full_unstemmed |
Media composition to revive leptospires stored at −80 °C |
title_sort |
media composition to revive leptospires stored at −80 °c |
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Academic Press |
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2022 |
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http://psasir.upm.edu.my/id/eprint/102162/ https://www.sciencedirect.com/science/article/pii/S0011224022000992?via%3Dihub |
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