title: Antihistomonal effects of artemisinin and Artemisia annua extracts in vitro could not be confirmed by in vivo experiments in turkeys and chickens.
abstract: Five different Artemisia annua-derived materials (i.e. dry leaves, pure artemisinin, and hexane, dichloromethane or methanol extracts of leaves) were screened for their in vitro activities against six clonal cultures of Histomonas meleagridis. Except for the methanol extract, all tested materials displayed in vitro activity against all tested protozoal clones. Neither the dry plant material, extracts nor artemisinin showed any antibacterial activity against the xenic bacteria accompanying the six H. meleagridis clones at concentration levels identical to the antihistomonal setting. The dichloromethane extract of dry leaves (Ext-DCM) (minimal lethal concentration=1.0 mg/ml) and artemisinin (half-maximal inhibitory concentration=1.295 mg/ml) had the most promising antihistomonal properties and were therefore subsequently tested in a standardized experimental infection model in both turkeys and chickens infected with clonal H. meleagridis. There were no differences between treatment groups, where all infected turkeys showed severe clinical histomonosis and demonstrated severe typhlohepatitis typical for histomonosis. Consistent with the infection model used, the infected chickens did not show any adverse clinical signs but contracted severe lesions in their caeca 7 and 10 days post infection (d.p.i.), liver lesions were absent to mild after 7 d.p.i. and progressed to severe lesions at 10 d.p.i.; thus no differences between treatment groups were observed. In conclusion, neither artemisinin nor Ext-DCM was able to prevent experimental histomonosis in turkeys and chickens at the given concentrations, which is contrary to the antihistomonal effect noticed in vitro even though the same clonal culture was used. The results of this study therefore clearly demonstrate the importance of defined in vivo experimentation in order to assess and verify in vitro results.
authors.affiliation: Department of Veterinary Disease Biology, Faculty of Health and Medical Science, University of Copenhagen, Frederiksberg C, Denmark. icnt@sund.ku.dk
authors.forename: I C N
authors.initials: IC
authors.lastname: Thøfner
authors.forename: D
authors.initials: D
authors.lastname: Liebhart
authors.forename: M
authors.initials: M
authors.lastname: Hess
authors.forename: T W
authors.initials: TW
authors.lastname: Schou
authors.forename: C
authors.initials: C
authors.lastname: Hess
authors.forename: E
authors.initials: E
authors.lastname: Ivarsen
authors.forename: X C
authors.initials: XC
authors.lastname: Fretté
authors.forename: L P
authors.initials: LP
authors.lastname: Christensen
authors.forename: K
authors.initials: K
authors.lastname: Grevsen
authors.forename: R M
authors.initials: RM
authors.lastname: Engberg
authors.forename: J P
authors.initials: JP
authors.lastname: Christensen
mesh_terms: D000818:Animals; D000890:Anti-Infective Agents / Q000494:pharmacology* / Q000627:therapeutic use; D031172:Artemisia annua / Q000737:chemistry*; D037621:Artemisinins / Q000494:pharmacology / Q000627:therapeutic use; D002645:Chickens; D004305:Dose-Response Relationship, Drug; D005260:Female; D020128:Inhibitory Concentration 50; D008099:Liver / Q000469:parasitology / Q000473:pathology; D008297:Male; D021261:Parasitic Sensitivity Tests; D010936:Plant Extracts / Q000737:chemistry / Q000494:pharmacology* / Q000627:therapeutic use; D018515:Plant Leaves / Q000737:chemistry; D011201:Poultry Diseases / Q000188:drug therapy* / Q000469:parasitology; D011529:Protozoan Infections, Animal / Q000188:drug therapy* / Q000469:parasitology; D011897:Random Allocation; D016834:Trichomonadida / Q000187:drug effects*; D014422:Turkeys
chemical_list: D000890:Anti-Infective Agents; D037621:Artemisinins; D010936:Plant Extracts; C031327:artemisinin