Differential ABC transporter gene expression in adult Dirofilaria immitis males and females following in vitro treatment with ivermectin, doxycycline or a combination of both
Abstract Background Combination doxycycline/macrocyclic lactone (ML) protocols have been shown to provide a more rapid adulticidal and microfilaricidal effect than either MLs or doxycycline alone, although female worms were reported to have a higher tolerance to treatments compared to male worms. Th...
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doaj-2315df5c475e49a28cfaab6919dfac402020-11-25T03:39:13ZengBMCParasites & Vectors1756-33052019-08-011211910.1186/s13071-019-3645-yDifferential ABC transporter gene expression in adult Dirofilaria immitis males and females following in vitro treatment with ivermectin, doxycycline or a combination of bothChiara Lucchetti0Marco Genchi1Luigi Venco2Alessandro Menozzi3Paolo Serventi4Simone Bertini5Chiara Bazzocchi6Laura Helen Kramer7Alice Vismarra8Dept. of Veterinary Sciences, University of Parma, Parasitology UnitDept. of Veterinary Sciences, University of Parma, Parasitology UnitClinica Veterinaria Lago MaggioreDept. of Veterinary Sciences, University of Parma, Pharmacology UnitDept. of Veterinary Sciences, University of Parma, Pharmacology UnitDept. of Veterinary Sciences, University of Parma, Pharmacology UnitDept. of Veterinary Science, University of MilanDept. of Veterinary Sciences, University of Parma, Parasitology UnitDept. of Veterinary Sciences, University of Parma, Parasitology UnitAbstract Background Combination doxycycline/macrocyclic lactone (ML) protocols have been shown to provide a more rapid adulticidal and microfilaricidal effect than either MLs or doxycycline alone, although female worms were reported to have a higher tolerance to treatments compared to male worms. The present study aimed to evaluate how ABC transporters may be involved in the synergic effect of the combination treatment. Adult worms of D. immitis were treated in vitro for 24 hours with doxycycline (DOXY), ivermectin (IVM) and a combination of both, and changes in the modulation of ABC transporter genes were measured. Levels of doxycycline inside different treatment media, post-treatment, were determined through HPLC analysis. Results Quantitative RT-PCR analysis showed the presence of changes in the modulation of ABC transporter genes evaluated in this study. In particular, in female worms, the combination treatment induced a substantial increase in gene expressions, especially of Dim-pgp-10 and Dim-haf-4; whereas in male worms, the greatest increase in gene expression was observed for Dim-pgp-10 and Dim-pgp-11 when treated with DMSO + IVM and DMSO + DOXY/IVM. HPLC analysis of the DOXY concentrations in the media after in vitro treatments of male worms showed a slight difference between the DMSO + DOXY samples and the combination (DMSO + DOXY + IVM), while no difference was observed among females. Conclusions Further studies are required to explain whether the modulation of cellular efflux plays a role, even partially, in the adulticide effect of doxycycline/macrocyclic lactone combinations in heartworm-infected dogs. To the authors’ knowledge, this is the first study to evaluate P-gp expression in adult D. immitis.http://link.springer.com/article/10.1186/s13071-019-3645-yDirofilaria immitisPgPGene expressionDoxycyclineIvermectinCombined treatment |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Chiara Lucchetti Marco Genchi Luigi Venco Alessandro Menozzi Paolo Serventi Simone Bertini Chiara Bazzocchi Laura Helen Kramer Alice Vismarra |
spellingShingle |
Chiara Lucchetti Marco Genchi Luigi Venco Alessandro Menozzi Paolo Serventi Simone Bertini Chiara Bazzocchi Laura Helen Kramer Alice Vismarra Differential ABC transporter gene expression in adult Dirofilaria immitis males and females following in vitro treatment with ivermectin, doxycycline or a combination of both Parasites & Vectors Dirofilaria immitis PgP Gene expression Doxycycline Ivermectin Combined treatment |
author_facet |
Chiara Lucchetti Marco Genchi Luigi Venco Alessandro Menozzi Paolo Serventi Simone Bertini Chiara Bazzocchi Laura Helen Kramer Alice Vismarra |
author_sort |
Chiara Lucchetti |
title |
Differential ABC transporter gene expression in adult Dirofilaria immitis males and females following in vitro treatment with ivermectin, doxycycline or a combination of both |
title_short |
Differential ABC transporter gene expression in adult Dirofilaria immitis males and females following in vitro treatment with ivermectin, doxycycline or a combination of both |
title_full |
Differential ABC transporter gene expression in adult Dirofilaria immitis males and females following in vitro treatment with ivermectin, doxycycline or a combination of both |
title_fullStr |
Differential ABC transporter gene expression in adult Dirofilaria immitis males and females following in vitro treatment with ivermectin, doxycycline or a combination of both |
title_full_unstemmed |
Differential ABC transporter gene expression in adult Dirofilaria immitis males and females following in vitro treatment with ivermectin, doxycycline or a combination of both |
title_sort |
differential abc transporter gene expression in adult dirofilaria immitis males and females following in vitro treatment with ivermectin, doxycycline or a combination of both |
publisher |
BMC |
series |
Parasites & Vectors |
issn |
1756-3305 |
publishDate |
2019-08-01 |
description |
Abstract Background Combination doxycycline/macrocyclic lactone (ML) protocols have been shown to provide a more rapid adulticidal and microfilaricidal effect than either MLs or doxycycline alone, although female worms were reported to have a higher tolerance to treatments compared to male worms. The present study aimed to evaluate how ABC transporters may be involved in the synergic effect of the combination treatment. Adult worms of D. immitis were treated in vitro for 24 hours with doxycycline (DOXY), ivermectin (IVM) and a combination of both, and changes in the modulation of ABC transporter genes were measured. Levels of doxycycline inside different treatment media, post-treatment, were determined through HPLC analysis. Results Quantitative RT-PCR analysis showed the presence of changes in the modulation of ABC transporter genes evaluated in this study. In particular, in female worms, the combination treatment induced a substantial increase in gene expressions, especially of Dim-pgp-10 and Dim-haf-4; whereas in male worms, the greatest increase in gene expression was observed for Dim-pgp-10 and Dim-pgp-11 when treated with DMSO + IVM and DMSO + DOXY/IVM. HPLC analysis of the DOXY concentrations in the media after in vitro treatments of male worms showed a slight difference between the DMSO + DOXY samples and the combination (DMSO + DOXY + IVM), while no difference was observed among females. Conclusions Further studies are required to explain whether the modulation of cellular efflux plays a role, even partially, in the adulticide effect of doxycycline/macrocyclic lactone combinations in heartworm-infected dogs. To the authors’ knowledge, this is the first study to evaluate P-gp expression in adult D. immitis. |
topic |
Dirofilaria immitis PgP Gene expression Doxycycline Ivermectin Combined treatment |
url |
http://link.springer.com/article/10.1186/s13071-019-3645-y |
work_keys_str_mv |
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