A cysteine protease from the latex of Ficus benjamina has in vitro anthelmintic activity against Haemonchus contortus

Abstract Haemonchus contortus is a gastrointestinal nematode that is responsible for high mortality rates in ruminant herds. The resistance of nematodes to synthetic anthelmintics is widespread and requires a continuous search for new bioactive molecules, such as proteins. The objective of this study was to evaluate the anthelmintic potential of a protease purified from the latex of Ficus benjamina against H. contortus . Fresh latex was collected from plants via small incisions in the green stems, the rubber was removed by centrifugation, and the latex protein extract (LPE) was obtained. After LPE fractionation with ammonium sulfate and chromatography of the fraction containing the highest proteolytic activity on CM-cellulose, a cysteine protease (FbP) was purified. FbP has a molecular mass of approximately 23.97 kDa, and its proteolytic activity was stable between pH 6.0 and pH 10 and over a broad temperature range, with optimum activity at 60 °C. FbP inhibited both the development and exsheathment of H. contortus larvae, with 50% effective concentrations of 0.26 and 0.79 mg/mL, respectively. We conclude that this cysteine protease from F. benjamina latex with anthelmintic activity against H. contortus could be a promising alternative for the development of products for use in parasite control programmes.

Download Full-text

Evaluation of anthelmintic activity of biologically synthesized silver nanoparticles against the gastrointestinal nematode, Haemonchus contortus

Journal of Helminthology ◽

10.1017/s0022149x16000444 ◽

2016 ◽

Vol 91 (4) ◽

pp. 454-461 ◽

Cited By ~ 9

Author(s):

R.S. Tomar ◽

S. Preet

Keyword(s):

Electron Microscopy ◽

Silver Nanoparticles ◽

Haemonchus Contortus ◽

Anthelmintic Activity ◽

Gastrointestinal Nematode ◽

Experimental Plant ◽

Egg Hatch ◽

Transmission Electron ◽

Motility Inhibition

AbstractThe present study focuses on the in vitro anthelmintic activity of silver nanoparticles (AgNPs) synthesized using the aqueous extract of Azadirachta indica against Haemonchus contortus. The synthesized AgNPs were characterized by ultraviolet–visible (UV-Vis) spectrophotometry, scanning electron microscopy (SEM), transmission electron microscopy (TEM) and X-ray diffraction (XRD) studies. The UV-Vis spectrum exhibited a sharp peak at 420 nm, which was validated by electron microscopy, indicating the preparation of spherical nanoparticles measuring 15–25 nm in size. The in vitro study was based on an egg hatch assay (EHA) and adult motility inhibition assays. Six concentrations of AgNPs were used for EHA, ranging from 0.00001 to 1.0 μg/ml, and a range of 1–25 μg/ml was used for adult worms. The highest concentration induced 85 ± 2.89% egg hatch inhibition. The IC50 value for EHA was 0.001 μg/ml, whereas in vitro adult H. contortus motility inhibition was produced at 7.89 μg/ml (LC50). The effectiveness of A. indica leaf extract (aqueous) was also evaluated, which showed an IC50 value for EHA of 115.67 μg/ml, while the LC50 against adult H. contortus was 588.54 μg/ml. The overall findings of the present study show that the experimental plant extract contains reducing properties for the synthesis of AgNPs which, in turn, showed potent anthelmintic properties. This is the first report where AgNPs have been tested for their anthelmintic properties in an in vitro model.

Download Full-text

In vitro studies of the anthelmintic activity of Picrolemma sprucei Hook. f. (Simaroubaceae)

Acta Amazonica ◽

10.1590/s0044-59672006000300006 ◽

2006 ◽

Vol 36 (3) ◽

pp. 327-330 ◽

Cited By ~ 13

Author(s):

Rita de Cássia Saraiva Nunomura ◽

Ellen Cristina Costa da Silva ◽

Denilson Ferreira Oliveira ◽

Adriana Mello Garcia ◽

Jankerle Neves Boeloni ◽

...

Keyword(s):

Haemonchus Contortus ◽

Anthelmintic Activity ◽

Gastrointestinal Nematode ◽

Mortality Rates ◽

In Vitro Studies ◽

Wild Ruminants ◽

Ethanol Extracts ◽

First Time ◽

Gastrointestinal Nematode Parasite

1300 ppm (1.3 g / L), water and ethanol extracts prepared from stems or roots of Picrolemma sprucei Hook. f. were lethal (85-90 % mortality) in vitro to Haemonchus contortus (Barber Pole Worm) larvae, a gastrointestinal nematode parasite found in domestic and wild ruminants. Neosergeolide and isobrucein B were isolated in 0.0083 and 0.0070 % yield from dry, ground P. sprucei stems (0.89 kg). Neosergeolide, isobrucein B and the anthelmintic drug standard levamisole all caused comparable mortality rates (68-77 %) in vitro to H. contortus at similar concentrations (81-86 ppm). The anthelmintic activity of P. sprucei infusions (teas), alcohol extracts, and neosergeolide and isobrucein B, has therefore been demonstrated for the first time.

Download Full-text

Chemical characterization and in vitro anthelmintic activity of Citrus bergamia Risso and Citrus X paradisii Macfad essential oil against Haemonchus contortus Kirby isolate

Acta Tropica ◽

10.1016/j.actatropica.2021.105869 ◽

2021 ◽

Vol 217 ◽

pp. 105869

Author(s):

Vivien Patricia Garbin ◽

Beatriz Munguía ◽

Jenny Carolina Saldaña ◽

Cícero Deschamps ◽

Roger Raupp Cipriano ◽

...

Keyword(s):

Essential Oil ◽

Haemonchus Contortus ◽

Chemical Characterization ◽

Anthelmintic Activity ◽

Citrus Bergamia

Download Full-text

In vitro anthelmintic activity of Dennettia tripetala G. Baker (Annonaceae) Fruits against Haemonchus contortus

Journal of Parasitic Diseases ◽

10.1007/s12639-021-01438-2 ◽

2021 ◽

Author(s):

Rachel A. Nwosu ◽

Mohammed M. Suleiman ◽

Hussaina J. Makun ◽

Matthew P. Ameh ◽

Manji A. Shetshak ◽

...

Keyword(s):

Haemonchus Contortus ◽

Anthelmintic Activity ◽

Dennettia Tripetala

Download Full-text

In Vitro Nematocidal Effect and Anthelmintic Activity of Artemisia cina Against Haemonchus contortus in Gerbils and Relative Expression of Hc29 Gene in Transitional Larvae (L3–L4)

Acta Parasitologica ◽

10.1007/s11686-021-00364-w ◽

2021 ◽

Author(s):

Rosa Isabel Higuera-Piedrahita ◽

Mariana Dolores-Hernández ◽

Luis Gerardo Jiménez-Pérez ◽

Brígida C. Camacho-Enríquez ◽

Alejandro Zamilpa ◽

...

Keyword(s):

Haemonchus Contortus ◽

Anthelmintic Activity ◽

Relative Expression

Download Full-text

Anthelmintic activity in vitro of Citrus sinensis and Melaleuca quinquenervia essential oil from Cuba on Haemonchus contortus

Industrial Crops and Products ◽

10.1016/j.indcrop.2015.07.056 ◽

2015 ◽

Vol 76 ◽

pp. 647-652 ◽

Cited By ~ 7

Author(s):

Yousmel Aleman Gaínza ◽

Luciana Ferreira Domingues ◽

Oriela Pino Perez ◽

Márcio Dias Rabelo ◽

Eugenio Roque López ◽

...

Keyword(s):

Essential Oil ◽

Haemonchus Contortus ◽

Citrus Sinensis ◽

Anthelmintic Activity ◽

Melaleuca Quinquenervia

Download Full-text

The effects of flubendazole and its metabolites on the larval development of Haemonchus contortus (Nematoda: Trichostrongylidae): an in vitro study

Helminthologia ◽

10.2478/s11687-010-0042-7 ◽

2010 ◽

Vol 47 (4) ◽

pp. 269-272 ◽

Cited By ~ 6

Author(s):

H. Bártíková ◽

L. Skálová ◽

J. Lamka ◽

B. Szotáková ◽

M. Várady

Keyword(s):

Larval Development ◽

Haemonchus Contortus ◽

Anthelmintic Activity ◽

In Vitro Study ◽

Lethal Concentration ◽

Resistant Isolate ◽

Susceptible Isolate ◽

Marked Resistance ◽

Larval Development Test

AbstractThe anthelmintic effects of flubendazole (FLU), its two main metabolites reduced flubendazole (FLU-R) and hydrolyzed flubendazole (FLU-H), and thiabendazole (TBZ) were compared using an in vitro larval development test in two isolates of Haemonchus contortus, a fully susceptible isolate (HCS) and a multi-resistant isolate (HCR). Results were quantified as 50 % lethal concentration (LC50), 99 % lethal concentration (LC99), efficacy factor (EF), and resistance factor (RF). For HCS, both LC50 and LC99 of FLU were lower than those of the reference TBZ. The anthelmintic activity of FLU-R in HCS and HCR was 13 and 6 times lower than the activity of FLU, respectively. The anthelmintic activity of FLU-H was negligible (approximately 363–853 times lower) compared to that of FLU. Although a marked resistance of the HCR isolate to TBZ was confirmed, only a low tolerance to FLU-R and slightly higher tolerance to FLU were found.

Download Full-text

Natural Compounds from the Marine Brown Alga Caulocystis cephalornithos with Potent In Vitro-Activity against the Parasitic Nematode Haemonchus contortus

Pathogens ◽

10.3390/pathogens9070550 ◽

2020 ◽

Vol 9 (7) ◽

pp. 550

Author(s):

Aya C. Taki ◽

Robert Brkljača ◽

Tao Wang ◽

Anson V. Koehler ◽

Guangxu Ma ◽

...

Keyword(s):

Haemonchus Contortus ◽

Brown Alga ◽

Parasitic Nematode ◽

Anthelmintic Activity ◽

Adult Males ◽

Terrestrial Plants ◽

Third Stage ◽

Future Work ◽

Marine Brown Alga

Eight secondary metabolites (1 to 8) were isolated from a marine sponge, a marine alga and three terrestrial plants collected in Australia and subsequently chemically characterised. Here, these natural product-derived compounds were screened for in vitro-anthelmintic activity against the larvae and adult stages of Haemonchus contortus (barber’s pole worm)—a highly pathogenic parasitic nematode of ruminants. Using an optimised, whole-organism screening system, compounds were tested on exsheathed third-stage larvae (xL3s) and fourth-stage larvae (L4s). Anthelmintic activity was initially evaluated on these stages based on the inhibition of motility, development and/or changes in morphology (phenotype). We identified two compounds, 6-undecylsalicylic acid (3) and 6-tridecylsalicylic acid (4) isolated from the marine brown alga, Caulocystis cephalornithos, with inhibitory effects on xL3 and L4 motility and larval development, and the induction of a “skinny-straight” phenotype. Subsequent testing showed that these two compounds had an acute nematocidal effect (within 1–12 h) on adult males and females of H. contortus. Ultrastructural analysis of adult worms treated with compound 4 revealed significant damage to subcuticular musculature and associated tissues and cellular organelles including mitochondria. In conclusion, the present study has discovered two algal compounds possessing acute anthelmintic effects and with potential for hit-to-lead progression. Future work should focus on undertaking a structure-activity relationship study and on elucidating the mode(s) of action of optimised compounds.

Download Full-text

In Vitro Anthelmintic Activity of Crude Extracts of Artemisia herba-alba and Punica granatum against Haemonchus contortus

Journal of Parasitology Research ◽

10.1155/2020/4950196 ◽

2020 ◽

Vol 2020 ◽

pp. 1-7 ◽

Cited By ~ 3

Author(s):

Aliyi Hassen Ahmed ◽

Mebrat Ejo ◽

Teka Feyera ◽

Dereje Regassa ◽

Bahar Mummed ◽

...

Keyword(s):

Haemonchus Contortus ◽

Anthelmintic Activity ◽

Punica Granatum ◽

Limiting Factor ◽

Egg Hatching ◽

Traditional Use ◽

Egg Hatch ◽

Inhibition Effect ◽

Crude Extracts

Gastrointestinal nematodes (GINs) are the major limiting factor for the successfulness of livestock production throughout the world. Emergence of resistance strains as well as scarcity and high cost of the currently available drugs has led to the evaluation of other alternative helminth control options, mainly from plants. The current study is aimed at investigating the in vitro anthelmintic efficacy of crude methanolic extracts of two traditionally important medicinal plants, Artemisia herba-alba and Punica granatum, against Haemonchus contortus using adult motility assay (AMA) and egg hatch inhibition assay (EHIA). Four graded concentrations of the extracts were tested for both the AMA (10, 5, 2.5, and 1.25 mg/mg) and EHIA (0.1, 0.25, 0.5, and 1 mg/mL) in replicates. Albendazole and phosphate-buffered saline (AMA) or distilled water (EHIA) were used as the positive and negative controls, respectively. The crude extracts of A. herba-alba and P. granatum exhibited a potential anthelmintic activity at all dose levels in a concentration- and time-dependent fashion. The highest concentration (10 mg/mL) of all the extracts caused a significantly (p<0.05) superior nematocidal activity compared to the negative control. Moreover, significant and concentration-dependent egg hatching inhibition effect was observed from both plant extracts. Maximal (98.67%) egg hatching inhibition effect was exhibited by the flower extract of A. herba-alba at 1 mg/mL concentration. The relative egg hatch inhibition efficacy indicated that both plants caused a significantly (p<0.05) greater egg hatch inhibition within 48 hr of exposure. The current study validated the traditional use of both plants as a natural anthelmintic against H. contortus justifying a need to undertake detail pharmacological and toxicological investigation on both plants.

Download Full-text