Anthelmintic screening of five palmatilobae species used in Traditional Medicine in Katanga Province (DR Congo)

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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,

mavungug@unilu.ac.cd

;

Gael.NZUZIMAVUNGU@student.umons.ac.be

Background: Resistance to known anthelmintics has become a growing concern

1 _{. 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 madiensis subsp.

Vitex mombassae Vatke

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

In hi bi tio n ra te (% ) DMSO O. da ctylop hylla V. do niana V. fis cheri V. ma diens is 3-5 V. ma diens is 5 V. mo mbas sae 3 -5 V. mo mbas sae 5 Leva miso le 0 50 100 150

*

A

DCM leaves extracts In hi bi tio n ra te (% ) DMSO O. dacty lophy lla V. do niana V. fis cheri V. ma diens is 3-5 V. ma diens is 5 V. mo mbas sae 3 -5 V. mo mbas sae 5 Levami sole 0 50 100 150

B

**

*

EtOAc leaves extracts

In hi bi tio n ra te (% ) DMSO O. da ctylop hylla V. do niana V. fis cheri V. ma diens is 3-5 V. ma diens is 5 V. mo mbas sae 3 -5 V. mo mbas sae 5 Leva misole 0 50 100 150

*

C

MeOH leaves exctracts

*

D

Introduction

Conclusion

and

perspectives

References

Acknowledgements

The 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

(WMicrotracker

TM

_{ONE, Phylumtech). This system measured}

_{C. elegans movement in microtiter}

plates. This allowed for identifying molecules that affect worm movement

2

_{. 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

In hi bi tio n ra te (% ) DMSO O. dacty lophy lla V. do niana V. fis cheri V. ma diens is 3-5 V. ma diens is 5 V. mo mbas sae 3 -5 V. mo mbas sae 5 Levami sole 0 50 100 150

E

DCM stem barks extracts

In hi bi tio n ra te (% ) DMSO O. dacty lophy lla V. do niana V. fis cheri V. ma diens is 3-5 V. ma diens is 5 V. mo mbas sae 3 -5 V. mo mbas sae 5 Leva misole 0 50 100 150 F

EtOAc stem barks extracts

In hi bi tio n ra te (% ) DMSO O. da ctylop hylla V. do niana V. fis cheri V. ma diens is 3-5 V. ma diens is 5 V. mo mbas sae 3 -5 V. mo mbas sae 5 Levami sole 0 50 100 150

G

MeOH stem barks exctracts

In hi bi tio n ra te (% ) DMSO O. dacty lophy lla V. do niana V. fisch eri V. madie nsis 3-5 V. ma diens is 5 V. mo mbas sae 3 -5 V. momb assa e 5 Levami sole -50 0 50 100 150 H *

n-heptane root barks exctracts

In hi bi tio n ra te (% ) DMSO O. dacty lophy lla V. do niana V. fis cheri V. ma diens is 3-5 V. ma diens is 5 V. mo mbas sae 3-5 V. mo mbas sae 5 Levami sole 0 50 100 150

I

DCM root barks extracts

In hi bi tio n ra te (% ) DMSO O. da ctylop hylla V. do niana V. fis cheri V. ma diens is 3-5 V. ma diens is 5 V. mo mbas sae 3-5 V. mo mbas sae 5 Levami sole -50 0 50 100 150 J

EtOAc root barks extracts

In hi bi tio n ra te (% ) DMSO O. dacty lophy lla V. do niana V. fis cheri V. ma diens is 3-5 V. ma diens is 5 V. mo mbas sae 3 -5 V. mo mbas sae 5 Leva miso le 0 50 100 150

K

MeOH root barks exctracts