Evaluation of Anthelmintic Activity and Composition of Pumpkin (Cucurbita pepo L.) Seed Extracts-In Vitro and in Vivo Studies.
A significant number of studies report growing resistance in nematodes thriving in both humans and livestock. This study was conducted to evaluate the in vitro and in vivo anthelmintic efficiency of Curcubita pepo (C. pepo) L. hot water extract (HWE), cold water extract (CWE) or ethanol extract (ETE) on two model nematodes: Caenorhabditis elegans (C. elegans) and Heligmosoides bakeri (H. bakeri).


METHODS
Raman, IR and LC-MS spectroscopy analyses were performed on the studied plant material to deliver qualitative and quantitative data on the composition of the obtained extracts: ETE, HWE and CWE. The in vitro activity evaluation showed an impact of C. pepo extracts on C. elegans and different developmental stages of H. bakeri. The following in vivo experiments on mice infected with H. bakeri confirmed inhibitory properties of the most active pumpkin extract selected by the in vitro study. All of the extracts were found to contain cucurbitine, aminoacids, fatty acids, and-for the first time-berberine and palmatine were identified. All C. pepo seed extracts exhibited a nematidicidal potential in vitro, affecting the survival of L1 and L2 H. bakeri larvae. The ETE was the strongest and demonstrated a positive effect on H. bakeri eggs hatching and marked inhibitory properties against worm motility, compared to a PBS control. No significant effects of pumpkin seed extracts on C. elegans integrity or motility were found. The EtOH extract in the in vivo studies showed anthelmintic properties against both H. bakeri fecal egg counts and adult worm burdens. The highest egg counts reduction was observed for the 8 g/kg dose (IC50 against H. bakeri = 2.43; 95% Cl = 2.01-2.94). A decrease in faecal egg counts (FEC) was accompanied by a significant reduction in worm burden of the treated mice compared to the control group.


CONCLUSIONS
Pumpkin seed extracts may be used to control of Gastrointestinal (G.I.) nematode infections. This relatively inexpensive alternative to the currently available chemotherapeutic should be considered as a novel drug candidate in the nearest future.
Department of Parasitology and Invasive Diseases, Faculty of Veterinary Medicine, University of Life Sciences in Lublin, 12 Akademicka Street, 20-950 Lublin, Poland. grzybek.genetics@gmail.com.|Department of Molecular Biology, Institute of Genetics and Animal Breeding, Polish Academy of Science, Jastrzebiec, 05-552 Magdalenka, Poland. grzybek.genetics@gmail.com.
Maciej
M
Grzybek
Chair and Department of Pharmacognosy with Medicinal Plants Unit, Medical University of Lublin, 20-084 Lublin, Poland. virginia.kukula@gmail.com.
Wirginia
W
Kukula-Koch
Faculty of Biology and Animal Breeding, Department of Biological Basis of Animal Production, University of Life Sciences in Lublin, 20-950 Lublin, Poland. aneta.strachecka@up.lublin.pl.
Aneta
A
Strachecka
Faculty of Chemistry, Jagiellonian University, 30-060 Krakow, Poland. jaworska.aleksandra.maria@gmail.com.|Jagiellonian Centre for Experimental Therapeutics (JCET), Jagiellonian University, 30-348 Krakow, Poland. jaworska.aleksandra.maria@gmail.com.
Aleksandra
A
Jaworska
School of Life Sciences, University of Nottingham, Nottingham NG7 2RD, UK. andrew.phiri@unza.zm.|Department of Clinical Studies, School of Veterinary Medicine, University of Zambia, P.O. Box 32379 Lusaka, Zambia. andrew.phiri@unza.zm.
Andrew M
AM
Phiri
Department of Zoology, Animal Ecology & Wildlife Management, Faculty of Biology and Animal Breeding, University of Life Sciences in Lublin, 13 Akademicka Street, 20-950 Lublin, Poland. jerzy.paleolog@up.lublin.pl.
Jerzy
J
Paleolog
Department of Parasitology and Invasive Diseases, Faculty of Veterinary Medicine, University of Life Sciences in Lublin, 12 Akademicka Street, 20-950 Lublin, Poland. krzysztof.tomczuk@up.lublin.pl.
Krzysztof
K
Tomczuk
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000969:Antinematodal Agents / Q000737:chemistry / Q000494:pharmacology*; D001599:Berberine / Q000032:analysis; D001600:Berberine Alkaloids / Q000032:analysis; D028464:Cucurbita / Q000737:chemistry*; D005227:Fatty Acids / Q000032:analysis; D051379:Mice; D010936:Plant Extracts / Q000737:chemistry / Q000494:pharmacology*; D010940:Plant Proteins / Q000032:analysis; D017168:Rhabditida / Q000187:drug effects; D012639:Seeds / Q000737:chemistry*
D000596:Amino Acids; D000969:Antinematodal Agents; D001600:Berberine Alkaloids; D005227:Fatty Acids; D010936:Plant Extracts; D010940:Plant Proteins; C023494:cucurbitin protein, pumpkin; D001599:Berberine; C005413:palmatine
Caenorhabditis elegans; Cucurbitaceae; Heligmosoides bakeri; LC-ESI-MS; anthelmintics; berberine; in vivo studies; plant extracts; pumpkin; spectroscopy