The ATP bioluminescence assay: a new application and optimization for viability testing in the parasitic nematode Haemonchus contortus

Nguyen, Linh Thuy; Zajíčková, Markéta; Mašátová, Eva; Matoušková, Petra; Skálová, Lenka

doi:10.1186/s13567-021-00980-4

Cited by 11 publications

(18 citation statements)

References 28 publications

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“…The effect of SRT on H. contortus was ascertained by measuring ATP content in the adult worms [ 13 ]. Similarly, adults of H. contortus were incubated with the commonly used drugs MOP and LEV.…”

Section: Methodsmentioning

confidence: 99%

“…The samples were homogenized 30 s (6 m/s) in FastPrep-24 homogenizer (MP Biomedicals, Santa Ana, CA, USA) then centrifuged for 10 min (Thermo Scientific Biofuge Stratos, 13 200 rpm (16 978 × g )) at 4 °C. ATP level content was measured by the ATP Bioluminescence Assay Kit CLS II (Roche, Mannheim, Germany) according to the manufacturer’s protocol, with adjustments described in [ 13 ].…”

Section: Methodsmentioning

confidence: 99%

“…To eliminate variability in worm size in the adults, the ATP concentration was related to µg/mL of total proteins in a sample pellet. The protein was measured in technical duplicates per sample using bicinchoninic acid based on the manufacturer’s protocol (PierceTM BCA Protein Assay Kit, ThermoScientific), with adjustments described in [ 13 ].…”

Section: Methodsmentioning

confidence: 99%

“…Among these, drug repurposing is an interesting strategy based on testing drugs approved for the treatment of other diseases for anthelmintic activity. The major advantage of this strategy, also called “therapeutic switching”, is that a great deal of information about these drugs is already well known, and a new candidate may be made ready for clinical trials relatively quickly [ 13 ]. Over the past years, a variety of drug repurposing initiatives have been launched against many diseases, including helminthiases [ 13 ].…”

Section: Introductionmentioning

confidence: 99%

“…Two H. contortus strains with a different sensitivity to classical anthelmintic drugs were used in the study: ISE (Inbred-Susceptible-Edinburgh, MHco3) strain, which is sensitive to all classes of anthelmintics and IRE (Inbred-Resistant-Edinburgh; MHco5) strain with decreased sensitivity to ivermectin (IVM) and benzimidazole drugs [ 11 , 12 ]. Besides the classical egg hatch test, the effect of SRT in H. contortus adults was also monitored using an ATP bioluminescent assay [ 13 ]. The efficacy of SRT in H. contortus adults was compared with the commonly used anthelmintic drugs levamisole (LEV) and monepantel (MOP).…”

Section: Introductionmentioning

confidence: 99%

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Sertraline as a new potential anthelmintic against Haemonchus contortus: toxicity, efficacy, and biotransformation

Zajíčková

Prchal

Navrátilová

et al. 2021

Vet Res

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Haemonchus contortus is a parasitic nematode of ruminants which causes significant losses to many farmers worldwide. Since the drugs currently in use for the treatment of haemonchosis are losing their effectiveness due to the drug-resistance of this nematode, a new or repurposed drug is highly needed. As the antipsychotic drug sertraline (SRT) has been shown to be effective against the parasitic nematodes Trichuris muris, Ancylostoma caninum and Schistosoma mansoni, the aim of the present study was to evaluate the possible effect of SRT on H. contortus. The potential hepatotoxicity of SRT was tested in sheep, a common H. contortus host. In addition, the main metabolic pathways of SRT in H. contortus and the ovine liver were identified. While no effect of SRT on H. contortus egg hatching was observed, SRT was found to significantly decrease the viability of H. contortus adults in drug-sensitive and resistant strains, with its effect comparable to the commonly used anthelmintics levamisole and monepantel. Moreover, SRT in anthelmintically active concentrations showed no toxicity to the ovine liver. Biotransformation of SRT in H. contortus was weak, with most of the drug remaining unmetabolized. Production of the main metabolite hydroxy-SRT did not differ significantly between strains. Other minor metabolites such as SRT-O-glucoside, dihydroxy-SRT, and SRT-ketone were also identified in H. contorts adults. Compared to H. contortus, the ovine liver metabolized SRT more extensively, mainly via desmethylation and glucuronidation. In conclusion, the potency of SRT against H. contortus was proven, and it should be tested further toward possible repurposing.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Sertraline as a new potential anthelmintic against Haemonchus contortus: toxicity, efficacy, and biotransformation

Zajíčková

Prchal

Navrátilová

et al. 2021

Vet Res

Self Cite

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Special considerations for studies of extracellular vesicles from parasitic helminths: A community‐led roadmap to increase rigour and reproducibility

White

Sotillo

Ancarola

et al. 2023

J of Extracellular Vesicle

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Over the last decade, research interest in defining how extracellular vesicles (EVs) shape cross‐species communication has grown rapidly. Parasitic helminths, worm species found in the phyla Nematoda and Platyhelminthes, are well‐recognised manipulators of host immune function and physiology. Emerging evidence supports a role for helminth‐derived EVs in these processes and highlights EVs as an important participant in cross‐phylum communication. While the mammalian EV field is guided by a community‐agreed framework for studying EVs derived from model organisms or cell systems [e.g., Minimal Information for Studies of Extracellular Vesicles (MISEV)], the helminth community requires a supplementary set of principles due to the additional challenges that accompany working with such divergent organisms. These challenges include, but are not limited to, generating sufficient quantities of EVs for descriptive or functional studies, defining pan‐helminth EV markers, genetically modifying these organisms, and identifying rigorous methodologies for in vitro and in vivo studies. Here, we outline best practices for those investigating the biology of helminth‐derived EVs to complement the MISEV guidelines. We summarise community‐agreed standards for studying EVs derived from this broad set of non‐model organisms, raise awareness of issues associated with helminth EVs and provide future perspectives for how progress in the field will be achieved.

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Sensitivity of Haemonchus contortus to anthelmintics using different in vitro screening assays: a comparative study

et al. 2022

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Background Helminthiasis and resistance to commercial anthelmintic compounds are major causes of economic losses for livestock producers, resulting in an urgent need for new drugs and reliable in vitro screening tests capable of detecting potentially active products. Considering this, a series of novel benzimidazole derivatives (5-methylbenzimidazole 1,2-disubstituted, 5-carboxybenzimidazole, 5-methylbenzimidazole 2-one) was screened on exsheathed L3 (xL3) and on the adult stage of Haemonchus contortus (Kirby anthelmintic-susceptible McMaster isolate). Methods This work presents the set-up of an automated motility assay on the xL3 stage of H. contortus using an infrared tracking device (WMicrotracker One) together with a larval development test (xL3 to L4) and a motility assay on the adult stage of H. contortus. A comparative study of the sensitivity of these in vitro assays using commercial anthelmintics with different mechanisms of action was carried out, also evaluating anthelmintic activity of a series of novel benzimidazole derivatives. Results The automated xL3 assay had the great advantage of being able to analyze many compounds simultaneously, but it showed the limitation of having lower sensitivity, requiring higher concentrations of the commercial anthelmintics tested compared to those needed for the adult motility or development assays. Although none of the novel 1,2,5-tri-substituted benzimidazole derivatives could significantly decrease the motility of xL3s, one of them (1e) significantly affected the development of xL3s to L4, and five new compounds (1b, 1d, 1e, 2a and 2c) reduced the motility of H. contortus adult stage. Conclusions The analysis of the results strongly suggests that the in vitro xL3 to L4 development test, particularly for the L4 stage, could be closer to the pharmacological sensitivity of the adult stage of H. contortus (target of interest) for commercial anthelmintic selected, with different mechanisms of action, and for the series of benzimidazole derivatives assayed. Therefore, an automated motility assay on L4 using the infrared tracking device is being set up. Further studies will be conducted to evaluate the in vivo anthelmintic activity of the most active novel benzimidazole derivatives. Graphical Abstract

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The ATP bioluminescence assay: a new application and optimization for viability testing in the parasitic nematode Haemonchus contortus

Cited by 11 publications

References 28 publications

Sertraline as a new potential anthelmintic against Haemonchus contortus: toxicity, efficacy, and biotransformation

Sertraline as a new potential anthelmintic against Haemonchus contortus: toxicity, efficacy, and biotransformation

Special considerations for studies of extracellular vesicles from parasitic helminths: A community‐led roadmap to increase rigour and reproducibility

Sensitivity of Haemonchus contortus to anthelmintics using different in vitro screening assays: a comparative study

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