The ATP bioluminescence assay: a new application and optimization for viability testing in the parasitic nematode Haemonchus contortus
Abstract The parasitic gastrointestinal nematode Haemonchus contortus causes serious economic losses to agriculture due to infection and disease in small ruminant livestock. The development of new therapies requires appropriate viability testing, with methods nowadays relying on larval motility or d...
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doaj-fc14e386048f404089a1c535486f80da2021-10-03T11:07:28ZengBMCVeterinary Research1297-97162021-09-0152111010.1186/s13567-021-00980-4The ATP bioluminescence assay: a new application and optimization for viability testing in the parasitic nematode Haemonchus contortusLinh Thuy Nguyen0Markéta Zajíčková1Eva Mašátová2Petra Matoušková3Lenka Skálová4Department of Biochemical Sciences, Faculty of Pharmacy, Charles UniversityDepartment of Biochemical Sciences, Faculty of Pharmacy, Charles UniversityDepartment of Biochemical Sciences, Faculty of Pharmacy, Charles UniversityDepartment of Biochemical Sciences, Faculty of Pharmacy, Charles UniversityDepartment of Biochemical Sciences, Faculty of Pharmacy, Charles UniversityAbstract The parasitic gastrointestinal nematode Haemonchus contortus causes serious economic losses to agriculture due to infection and disease in small ruminant livestock. The development of new therapies requires appropriate viability testing, with methods nowadays relying on larval motility or development using procedures that involve microscopy. None of the existing biochemical methods, however, are performed in adults, the target stage of the anthelmintic compounds. Here we present a new test for the viability of H. contortus adults and exsheathed third-stage larvae which is based on a bioluminescent assay of ATP content normalized to total protein concentration measured using bicinchoninic acid. All the procedure steps were optimized to achieve maximal sensitivity and robustness. This novel method can be used as a complementary assay for the phenotypic screening of new compounds with potential antinematode activity in exsheathed third-stage larvae and in adult males. Additionally, it might be used for the detection of drug-resistant isolates.https://doi.org/10.1186/s13567-021-00980-4optimized protocollevamisoleanthelminticshelminthsadult wormsexsheathed third-stage larvae |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Linh Thuy Nguyen Markéta Zajíčková Eva Mašátová Petra Matoušková Lenka Skálová |
spellingShingle |
Linh Thuy Nguyen Markéta Zajíčková Eva Mašátová Petra Matoušková Lenka Skálová The ATP bioluminescence assay: a new application and optimization for viability testing in the parasitic nematode Haemonchus contortus Veterinary Research optimized protocol levamisole anthelmintics helminths adult worms exsheathed third-stage larvae |
author_facet |
Linh Thuy Nguyen Markéta Zajíčková Eva Mašátová Petra Matoušková Lenka Skálová |
author_sort |
Linh Thuy Nguyen |
title |
The ATP bioluminescence assay: a new application and optimization for viability testing in the parasitic nematode Haemonchus contortus |
title_short |
The ATP bioluminescence assay: a new application and optimization for viability testing in the parasitic nematode Haemonchus contortus |
title_full |
The ATP bioluminescence assay: a new application and optimization for viability testing in the parasitic nematode Haemonchus contortus |
title_fullStr |
The ATP bioluminescence assay: a new application and optimization for viability testing in the parasitic nematode Haemonchus contortus |
title_full_unstemmed |
The ATP bioluminescence assay: a new application and optimization for viability testing in the parasitic nematode Haemonchus contortus |
title_sort |
atp bioluminescence assay: a new application and optimization for viability testing in the parasitic nematode haemonchus contortus |
publisher |
BMC |
series |
Veterinary Research |
issn |
1297-9716 |
publishDate |
2021-09-01 |
description |
Abstract The parasitic gastrointestinal nematode Haemonchus contortus causes serious economic losses to agriculture due to infection and disease in small ruminant livestock. The development of new therapies requires appropriate viability testing, with methods nowadays relying on larval motility or development using procedures that involve microscopy. None of the existing biochemical methods, however, are performed in adults, the target stage of the anthelmintic compounds. Here we present a new test for the viability of H. contortus adults and exsheathed third-stage larvae which is based on a bioluminescent assay of ATP content normalized to total protein concentration measured using bicinchoninic acid. All the procedure steps were optimized to achieve maximal sensitivity and robustness. This novel method can be used as a complementary assay for the phenotypic screening of new compounds with potential antinematode activity in exsheathed third-stage larvae and in adult males. Additionally, it might be used for the detection of drug-resistant isolates. |
topic |
optimized protocol levamisole anthelmintics helminths adult worms exsheathed third-stage larvae |
url |
https://doi.org/10.1186/s13567-021-00980-4 |
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