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Anthelmintic screening of five palmatilobae species used in traditional medicine in Katanga Province (DR Congo)
Nzuzi M.G. 1,2,3, Muyumba N.W. 2,4, Gauthier M-L. 2, Panda S.K. 5, Luyten W. 5, Nachtergael A.2, Vandenput S.3, Bakari S.A.6, Okombe E.V.1 , Duez P.2
1 University of Lubumbashi (UNILU), Unit of Pharmacology and Therapeutic, Faculty of Veterinary Medicine, 1825, Lubumbashi, DR Congo
2 University of Mons (UMONS), Unit of Therapeutic Chemistry and Pharmacognosy, 7000 Mons, Belgium 3 University of Liège (Uliège), ULiège Library | Secteur des Sciences de la Santé, 4000 Liège, Belgium
4 Department of Chemistry-physics, High college training teachers of Lubumbashi, Lubumbashi, 1825, DR Congo 5 Department of Biology, Animal Physiology and Neurobiology Section, KU Leuven, Naamsestraat 59, box 2465, 3000 Leuven, Belgium
6 University of Lubumbashi (UNILU), Laboratory of Pharmacognosy, Faculty of Pharmacy, 1825, Lubumbashi, DR Congo
Corresponding autor : Nzuzi Mavungu Gaël, [email protected]; [email protected]
Background: Resistance to known anthelmintics has become a growing concern1. Based on 2017–2019 ethnomedical and ethnoveterinary surveys carried out in Katanga Province, several species of shrub locally identified as Vitex species (Lamiaceae) are used to treat animal helminthiases. Aim of the study: In this study, 4 Vitex species (including different morphotypes) and Oldfieldia dactylophylla (Welw. ex Oliv.) (Picrodendraceae; locally misidentified as Vitex congolensis De Wild and T. Durand) were selected for in vitro screening in order to search for new anthelmintic lead compounds.
Oldfieldia dactylophylla (Welw. ex Oliv.) Vitex doniana Sweet Vitex fischeri Gürke Vitex mombassae Vatke Vitex madiensis subsp. Milanjiensis (Britten) F.White
Methanol Ethyl acetate Dichloromethane n-heptane
R1 R2 B3 L4
We provide scientific support for the traditional use of 4 Vitex species and Oldfieldia dactylophylla in traditional medicine in Katanga province for treating intestinal parasites in veterinary practice. Anthelmintic screening study showed that 9/84 tested extracts are significantly active on Caenorhabditis elegans. The motility inhibitions measured on the N2 wild-type C. elegans model call for further studies to develop compounds or standardized extracts as possible treatments for parasitic worm infections.
1. Jackson, F., Varady, M., Bartley, D.J. 2012, Managing anthelmintic resistance in goats-Can we learn lessons from sheep. – Small Ruminant Research, 103, 3–9. 2. Liu, M., Veryser, C., Lu, J-G., Wenseleers, T., De Borggraeve, W.M., Jiang, Z-H., Luyten, W. 2018, Bioassay-guided isolation of active substances from Semen Torreyae identifies two new anthelmintic compounds with novel mechanism of action. – Journal of Ethnopharmacology, 224, 421–428.
n-heptane leaves extracts
Inhi
bitio
n ra
te (%
)
DMSO
O. dactylo
phylla
V. doniana
V. fischeri
V. madien
sis 3-5
V. madien
sis 5
V. mombass
ae 3-5
V. mombass
ae 5
Levamiso
le0
50
100
150
*
A
DCM leaves extracts
Inhi
bitio
n ra
te (%
)
DMSO
O. dactylo
phylla
V. doniana
V. fischeri
V. madien
sis 3-5
V. madien
sis 5
V. mombass
ae 3-5
V. mombass
ae 5
Levamiso
le0
50
100
150B
***
*
*
EtOAc leaves extracts
Inhi
bitio
n ra
te (%
)
DMSO
O. dactylo
phylla
V. doniana
V. fischeri
V. madien
sis 3-5
V. madien
sis 5
V. mombass
ae 3-5
V. mombass
ae 5
Levamiso
le0
50
100
150
*
C
MeOH leaves exctracts
Inhi
bitio
n ra
te (%
)
DMSO
O. dactylo
phylla
V. doniana
V. fischeri
V. madien
sis 3-5
V. madien
sis 5
V. mombass
ae 3-5
V. mombass
ae 5
Levamiso
le0
50
100
150
*
D
Introduction
Conclusion and perspectives
References
AcknowledgementsThe ARES is gratefully acknowledged for financial support through the Phytokat research project. We thank Claudio Palmieri and Kobe Lavaerts for their assistance in our laboratory work.
Caenorhabditis elegans motility screen The motility-based screening was performed using an infrared detection system (WMicrotrackerTM ONE, Phylumtech). This system measured C. elegans movement in microtiter plates. This allowed for identifying molecules that affect worm movement2. Levamisole was used as positive control.
NGM plate + E. coli OP50
Synchronised young adult C. elegans
Wash 50 worms / well
96-wells plate
- 189 µL of E.coli OP50 - 10 µl of worm solution - 1 µL of extract/Levamisole/DMSO
Life cycle of C. elegans
Rotary evaporator Extracs
Drying and milling of plants organs : leaves, stem barks and root barks
2 g of each powder percolated (maceration) into 20 mL of solvents with increasing polaritiesy for 24 h.
obtained macerates
n-heptane stem barks extracts
Inhi
bitio
n ra
te (%
)
DMSO
O. dactylo
phylla
V. doniana
V. fischeri
V. madien
sis 3-5
V. madien
sis 5
V. mombass
ae 3-5
V. mombass
ae 5
Levamiso
le0
50
100
150E
DCM stem barks extracts
Inhi
bitio
n ra
te (%
)
DMSO
O. dactylo
phylla
V. doniana
V. fischeri
V. madien
sis 3-5
V. madien
sis 5
V. mombass
ae 3-5
V. mombass
ae 5
Levamiso
le0
50
100
150F
EtOAc stem barks extracts
Inhi
bitio
n ra
te (%
)
DMSO
O. dactylo
phylla
V. doniana
V. fischeri
V. madien
sis 3-5
V. madien
sis 5
V. mombass
ae 3-5
V. mombass
ae 5
Levamiso
le0
50
100
150G
MeOH stem barks exctracts
Inhi
bitio
n ra
te (%
)
DMSO
O. dactylo
phylla
V. doniana
V. fischeri
V. madien
sis 3-5
V. madien
sis 5
V. mombass
ae 3-5
V. mombass
ae 5
Levamiso
le-50
0
50
100
150H
*
n-heptane root barks exctracts
Inhi
bitio
n ra
te (%
)
DMSO
O. dactylo
phylla
V. doniana
V. fischeri
V. madien
sis 3-5
V. madien
sis 5
V. mombass
ae 3-5
V. mombass
ae 5
Levamiso
le0
50
100
150I
DCM root barks extracts
Inhi
bitio
n ra
te (%
)
DMSO
O. dactylo
phylla
V. doniana
V. fischeri
V. madien
sis 3-5
V. madien
sis 5
V. mombass
ae 3-5
V. mombass
ae 5
Levamiso
le-50
0
50
100
150J
EtOAc root barks extracts
Inhi
bitio
n ra
te (%
)
DMSO
O. dactylo
phylla
V. doniana
V. fischeri
V. madien
sis 3-5
V. madien
sis 5
V. mombass
ae 3-5
V. mombass
ae 5
Levamiso
le0
50
100
150K
MeOH root barks exctracts
Inhi
bitio
n ra
te (%
)
DMSO
O. dactylo
phylla
V. doniana
V. fischeri
V. madien
sis 3-5
V. madien
sis 5
V. mombass
ae 3-5
V. mombass
ae 5
Levamiso
le0
50
100
150
*
L
