Pub Date : 2025-04-16DOI: 10.1016/j.ijpddr.2025.100595
Lúcia Mamede , Fanta Fall , Madeline Vast , Kristelle Hughes , Giorgia Martelli , Francesco Caligiore , Bernadette Govaerts , Paul A.M. Michels , Michel Frédérich , Joëlle Quetin-Leclercq
Trypanosomiasis is a parasitic disease for which new treatments are needed due to the frequent occurrence of adverse side effects of current available drugs. Natural compounds found in traditionally used plants offer opportunities to discover innovative compounds that could prove pivotal to antitrypanosomal drug development. 3-O-p-(Z/E)-coumaroyltormentic acids (CTA) were isolated first from the West Africa-native tree Vitellaria paradoxa and have demonstrated quite selective in vitro and in vivo antitrypanosomal activity, despite the unknown mode of action. In this study, a metabolomics analysis using the data from both LC-HR-MS and 1H-NMR described CTA's effects on Trypanosoma brucei after 3 h exposure under 5 or 10 x EC50. Our study shows CTA's activity impacted tryptophan metabolism and reveals potential targets in different branches of this metabolism. Our results demonstrate a likely presence of enzymes dedicated to tryptophan, like a tryptophan aminotransferase, tryptophan 2,3-dioxygenase and/or indoleamine 2,3-dioxygenase, and other enzymes of the kynurenine pathway, despite the absence of their description thus far in this species. These data further implicate that CTA's toxic effect on the tryptophan metabolism may be attributed to the decrease of the intracellular level of essential aspartate, resulting from inhibition of its aminotransferase. In resume, our study shines light on the likelihood of the tryptophan metabolism pathway presenting innovative targets toward the development of antitrypanosomal drugs. These require confirmation through functional and enzymatic studies.
{"title":"Metabolomics study of 3-O-p-(Z/E)-coumaroyltormentic acid-treated Trypanosoma brucei brucei","authors":"Lúcia Mamede , Fanta Fall , Madeline Vast , Kristelle Hughes , Giorgia Martelli , Francesco Caligiore , Bernadette Govaerts , Paul A.M. Michels , Michel Frédérich , Joëlle Quetin-Leclercq","doi":"10.1016/j.ijpddr.2025.100595","DOIUrl":"10.1016/j.ijpddr.2025.100595","url":null,"abstract":"<div><div>Trypanosomiasis is a parasitic disease for which new treatments are needed due to the frequent occurrence of adverse side effects of current available drugs. Natural compounds found in traditionally used plants offer opportunities to discover innovative compounds that could prove pivotal to antitrypanosomal drug development. 3-O-<em>p</em>-(<em>Z/E</em>)-coumaroyltormentic acids (CTA) were isolated first from the West Africa-native tree <em>Vitellaria paradoxa</em> and have demonstrated quite selective <em>in vitro</em> and <em>in vivo</em> antitrypanosomal activity, despite the unknown mode of action. In this study, a metabolomics analysis using the data from both LC-HR-MS and <sup>1</sup>H-NMR described CTA's effects on <em>Trypanosoma brucei</em> after 3 h exposure under 5 or 10 x EC<sub>50</sub>. Our study shows CTA's activity impacted tryptophan metabolism and reveals potential targets in different branches of this metabolism. Our results demonstrate a likely presence of enzymes dedicated to tryptophan, like a tryptophan aminotransferase, tryptophan 2,3-dioxygenase and/or indoleamine 2,3-dioxygenase, and other enzymes of the kynurenine pathway, despite the absence of their description thus far in this species. These data further implicate that CTA's toxic effect on the tryptophan metabolism may be attributed to the decrease of the intracellular level of essential aspartate, resulting from inhibition of its aminotransferase. In resume, our study shines light on the likelihood of the tryptophan metabolism pathway presenting innovative targets toward the development of antitrypanosomal drugs. These require confirmation through functional and enzymatic studies.</div></div>","PeriodicalId":13775,"journal":{"name":"International Journal for Parasitology: Drugs and Drug Resistance","volume":"28 ","pages":"Article 100595"},"PeriodicalIF":4.1,"publicationDate":"2025-04-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144070138","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-04-15DOI: 10.1016/j.ijpddr.2025.100592
Anne-Charlotte Lenière , Amit Upadhyay , Jérôme Follet , Timothy P. O'Sullivan
The protozoan parasite Cryptosporidium is the etiological agent of cryptosporidiosis, a ubiquitous diarrheic disease affecting humans and animals. Treatment options are limited, highlighting an urgent need for novel therapeutics. Despite decades of research and a wide diversity of strategies to tackle parasite metabolic pathways, no completely effective drug has been identified to date. Within targeted parasite enzymatic and metabolic pathways, the synthesis of nucleotide mediated by the inosine 5′-monophosphate dehydrogenase (IMPDH) enzyme is the focus of significant research efforts. Based on our prior studies of bacterial IMPDH inhibitors, we report herein the development and characterisation of novel inhibitors targeting Cryptosporidium parvum IMPDH (CpIMPDH). Specifically, we synthesised heteroaryl-containing urea and squaramide analogues to evaluate their potential in vitro anti-Cryptosporidium activity. Initial screening identified nine active compounds with the most potent candidates achieving IC50 values as low as 2.2 μM. Subsequent time-course experiments revealed that the molecules effectively inhibit parasite invasion and early intracellular development but failed to tackle C. parvum growth when introduced at 30 h post infection. The present work introduces a new family of squaramide-derived IMPDH inhibitors and also interrogates the need to standardise commonly accepted protocols used for assessing anti-cryptosporidial drug activity.
{"title":"Effect of urea and squaramide IMPDH inhibitors on C. parvum: in vitro trial design impacts the assessment of drug efficacy","authors":"Anne-Charlotte Lenière , Amit Upadhyay , Jérôme Follet , Timothy P. O'Sullivan","doi":"10.1016/j.ijpddr.2025.100592","DOIUrl":"10.1016/j.ijpddr.2025.100592","url":null,"abstract":"<div><div>The protozoan parasite <em>Cryptosporidium</em> is the etiological agent of cryptosporidiosis, a ubiquitous diarrheic disease affecting humans and animals. Treatment options are limited, highlighting an urgent need for novel therapeutics. Despite decades of research and a wide diversity of strategies to tackle parasite metabolic pathways, no completely effective drug has been identified to date. Within targeted parasite enzymatic and metabolic pathways, the synthesis of nucleotide mediated by the inosine 5′-monophosphate dehydrogenase (IMPDH) enzyme is the focus of significant research efforts. Based on our prior studies of bacterial IMPDH inhibitors, we report herein the development and characterisation of novel inhibitors targeting <em>Cryptosporidium parvum</em> IMPDH (<em>Cp</em>IMPDH). Specifically, we synthesised heteroaryl-containing urea and squaramide analogues to evaluate their potential <em>in vitro</em> anti-<em>Cryptosporidium</em> activity. Initial screening identified nine active compounds with the most potent candidates achieving IC<sub>50</sub> values as low as 2.2 μM. Subsequent time-course experiments revealed that the molecules effectively inhibit parasite invasion and early intracellular development but failed to tackle <em>C. parvum</em> growth when introduced at 30 h post infection. The present work introduces a new family of squaramide-derived IMPDH inhibitors and also interrogates the need to standardise commonly accepted protocols used for assessing anti-cryptosporidial drug activity.</div></div>","PeriodicalId":13775,"journal":{"name":"International Journal for Parasitology: Drugs and Drug Resistance","volume":"28 ","pages":"Article 100592"},"PeriodicalIF":4.1,"publicationDate":"2025-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143898678","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
This study explores the anti-proliferative and anti-parasitic properties of ursolic acid (UA) in Theileria annulata-infected bovine (TA) cells. Dose-response analyses determined an IC50 value of approximately 5 μg/mL for UA, demonstrating selective cytotoxicity toward infected cells with minimal impact on healthy cells. UA treatment induced pronounced morphological alterations and apoptosis in TA cells, as evidenced by light microscopy and a time-dependent increase in cell mortality. Notably, UA exhibited consistent efficacy against both buparvaquone (BPQ)-sensitive and BPQ-resistant TA cell lines, highlighting its broad-spectrum anti-parasitic potential. Mechanistic investigations revealed that UA triggers DNA damage, elevates reactive oxygen species (ROS) levels, disrupts mitochondrial function, and induces sub-G1 phase arrest, culminating in apoptosis primarily via the intrinsic pathway. Mass spectrometry-based proteomic profiling identified significant perturbations in host cell pathways, including DNA repair mechanisms, cell cycle regulation, and signaling networks, alongside direct interference with parasite metabolic processes. Western blot analysis further confirmed UA-mediated modulation of host cell signaling pathways and chromatin organization. Given the rising incidence of drug-resistant T. annulata strains, the development of novel therapeutic strategies is imperative. These findings highlight UA's multifaceted mechanism of action, targeting both parasitic and host cellular processes, and position it as a promising candidate for the treatment of bovine theileriosis.
{"title":"Ursolic acid induces apoptosis and disrupts host-parasite interactions in Theileria annulata-infected cells","authors":"Sakshi Singh , Madhusmita Subudhi , Vengatachala Moorthy A , Akash Suresh , Paresh Sharma","doi":"10.1016/j.ijpddr.2025.100593","DOIUrl":"10.1016/j.ijpddr.2025.100593","url":null,"abstract":"<div><div>This study explores the anti-proliferative and anti-parasitic properties of ursolic acid (UA) in <em>Theileria annulata</em>-infected bovine (TA) cells. Dose-response analyses determined an IC<sub>50</sub> value of approximately 5 μg/mL for UA, demonstrating selective cytotoxicity toward infected cells with minimal impact on healthy cells. UA treatment induced pronounced morphological alterations and apoptosis in TA cells, as evidenced by light microscopy and a time-dependent increase in cell mortality. Notably, UA exhibited consistent efficacy against both buparvaquone (BPQ)-sensitive and BPQ-resistant TA cell lines, highlighting its broad-spectrum anti-parasitic potential. Mechanistic investigations revealed that UA triggers DNA damage, elevates reactive oxygen species (ROS) levels, disrupts mitochondrial function, and induces sub-G1 phase arrest, culminating in apoptosis primarily via the intrinsic pathway. Mass spectrometry-based proteomic profiling identified significant perturbations in host cell pathways, including DNA repair mechanisms, cell cycle regulation, and signaling networks, alongside direct interference with parasite metabolic processes. Western blot analysis further confirmed UA-mediated modulation of host cell signaling pathways and chromatin organization. Given the rising incidence of drug-resistant <em>T. annulata</em> strains, the development of novel therapeutic strategies is imperative. These findings highlight UA's multifaceted mechanism of action, targeting both parasitic and host cellular processes, and position it as a promising candidate for the treatment of bovine theileriosis.</div></div>","PeriodicalId":13775,"journal":{"name":"International Journal for Parasitology: Drugs and Drug Resistance","volume":"28 ","pages":"Article 100593"},"PeriodicalIF":4.1,"publicationDate":"2025-04-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143860023","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-04-14DOI: 10.1016/j.ijpddr.2025.100594
Kaylee R. Kipp , Elizabeth M. Redman , Joe L. Luksovsky , Dani Claussen , John S. Gilleard , Guilherme G. Verocai
Bison production is a growing sector of the United States agriculture, with more consumers choosing bison products. Commercial bison are kept on smaller pastures and treated with anthelmintics for gastrointestinal nematodes (GIN) to maintain production. However, there is a lack of information regarding the GIN parasite communities in ranched bison or the extent of their resistance to anthelmintics. Our objectives were: i) to determine the GIN species present and the extent of resistance to the benzimidazole drug class in commercial bison herds in the southern US and ii) to assess age class differences in GIN species composition and BZ resistance. Composite coprocultures from bison in Texas (n = 14) and Oklahoma (n = 2), and individual bison of different age classes from a single ranch in central Texas (n = 43) were analyzed using ITS2 rDNA nemabiome metabarcoding to determine the trichostrongylid species composition. For both the composite and individual samples, the most common trichostrongylid species found were Haemonchus contortus, Haemonchus placei, and Ostertagia ostertagi. Among the known canonical isotype-1 β-tubulin BZ resistance polymorphisms (at codons 200, 198, 167), the 200Y (TTC > TAC) substitution was the most widespread across the two southern states, with a prevalence of 81.3 %. Other polymorphisms, such as 167Y (TTC > TAC) and 198L (GAA > TTA), were also detected, and both had prevalences of 62.5 %. Ostertagia ostertagi was found to have very high frequencies (overall mean frequency = 62.6 %; range = 28.3–100 %) of the 200Y (TTC > TAC) polymorphism in all age classes sampled. Overall, benzimidazole resistance polymorphisms were found at moderate to high frequency in the three major economically important GIN species in ranched bison in Texas and Oklahoma, suggesting a potential widespread distribution of benzimidazole resistance polymorphisms in the southern United States. This work has important implications for all other grazing livestock and illustrates the importance of early detection of anthelmintic resistance and the need for mitigation strategies.
{"title":"High frequency of benzimidazole resistance polymorphisms and age-class differences in trichostrongyle nematodes of ranched bison from the south-central United States","authors":"Kaylee R. Kipp , Elizabeth M. Redman , Joe L. Luksovsky , Dani Claussen , John S. Gilleard , Guilherme G. Verocai","doi":"10.1016/j.ijpddr.2025.100594","DOIUrl":"10.1016/j.ijpddr.2025.100594","url":null,"abstract":"<div><div>Bison production is a growing sector of the United States agriculture, with more consumers choosing bison products. Commercial bison are kept on smaller pastures and treated with anthelmintics for gastrointestinal nematodes (GIN) to maintain production. However, there is a lack of information regarding the GIN parasite communities in ranched bison or the extent of their resistance to anthelmintics. Our objectives were: i) to determine the GIN species present and the extent of resistance to the benzimidazole drug class in commercial bison herds in the southern US and ii) to assess age class differences in GIN species composition and BZ resistance. Composite coprocultures from bison in Texas (<em>n</em> = 14) and Oklahoma (<em>n</em> = 2), and individual bison of different age classes from a single ranch in central Texas (<em>n</em> = 43) were analyzed using ITS2 rDNA nemabiome metabarcoding to determine the trichostrongylid species composition. For both the composite and individual samples, the most common trichostrongylid species found were <em>Haemonchus contortus, Haemonchus placei</em>, and <em>Ostertagia ostertagi</em>. Among the known canonical isotype-1 β-tubulin BZ resistance polymorphisms (at codons 200, 198, 167), the 200Y (TTC > T<u>A</u>C) substitution was the most widespread across the two southern states, with a prevalence of 81.3 %. Other polymorphisms, such as 167Y (TTC > T<u>A</u>C) and 198L (GAA > <u>TT</u>A), were also detected, and both had prevalences of 62.5 %. <em>Ostertagia ostertagi</em> was found to have very high frequencies (overall mean frequency = 62.6 %; range = 28.3–100 %) of the 200Y (TTC > T<u>A</u>C) polymorphism in all age classes sampled. Overall, benzimidazole resistance polymorphisms were found at moderate to high frequency in the three major economically important GIN species in ranched bison in Texas and Oklahoma, suggesting a potential widespread distribution of benzimidazole resistance polymorphisms in the southern United States. This work has important implications for all other grazing livestock and illustrates the importance of early detection of anthelmintic resistance and the need for mitigation strategies.</div></div>","PeriodicalId":13775,"journal":{"name":"International Journal for Parasitology: Drugs and Drug Resistance","volume":"28 ","pages":"Article 100594"},"PeriodicalIF":4.1,"publicationDate":"2025-04-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143838341","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-04-14DOI: 10.1016/j.ijpddr.2025.100591
Fernanda G. Fumuso , Jason A. Clement , Matthew J. Todd , Roberta M. O'Connor
Toxoplasmosis is a worldwide parasitic disease caused by the apicomplexan Toxoplasma gondii. Severe neurological illness occurs in immunosuppressed patients, and congenital disorders can follow transplacental primo infection during pregnancy. New effective antiparasitic drugs are needed since chronic cystic stages are resistant to current available treatments, and some of the congenital infections are unresponsive to available therapeutics. Tartrolon E (trtE) is a marine secondary metabolite that has high selectivity against human and animal apicomplexan parasites including T. gondii, Cryptosporidium parvum and Plasmodium falciparum. We evaluated the effect of the compound on extracellular tachyzoite viability, morphology, membrane permeability and its ability to block host cell attachment and/or invasion. While 80 % of T. gondii infective capacity is blocked after only 30 min of compound treatment, parasite viability, morphology, membrane integrity and host cell attachment were unaffected until after 4 h of treatment. These effects were irreversible when parasites were allowed to infect host cells after trtE treatment. Drug exposure for more than 4 h significantly affected tachyzoite survival and altered parasite morphology. The mechanism of action of trtE is still unknown but includes blocking parasite invasion processes. Further research is needed to determine the molecular target of trtE to further progress the compound as an antiparasitic candidate.
弓形虫病是由弓形虫顶复体引起的一种世界性寄生虫病。严重的神经系统疾病发生在免疫抑制的患者中，先天性疾病可在怀孕期间发生经胎盘原发性感染。需要新的有效的抗寄生虫药物，因为慢性囊性病变对现有的治疗方法有抗药性，而且一些先天性感染对现有的治疗方法没有反应。Tartrolon E （trtE）是一种海洋次生代谢物，对人类和动物的顶复体寄生虫，包括弓形虫、小隐孢子虫和恶性疟原虫有很高的选择性。我们评估了该化合物对细胞外速殖子活力、形态、膜通透性及其阻止宿主细胞附着和/或入侵的能力的影响。虽然80%的弓形虫感染能力在复合处理30分钟后被阻断，但寄生虫的活力、形态、膜完整性和宿主细胞附着直到处理4小时后才受到影响。当寄生虫在trtE治疗后感染宿主细胞时，这些效应是不可逆的。药物暴露超过4小时显著影响速殖子存活和改变寄生虫形态。trtE的作用机制尚不清楚，但包括阻断寄生虫入侵过程。需要进一步研究确定trtE的分子靶点，以进一步开发该化合物作为抗寄生虫候选药物。
{"title":"Tartrolon E rapidly blocks Toxoplasma gondii capacity to invade host cells","authors":"Fernanda G. Fumuso , Jason A. Clement , Matthew J. Todd , Roberta M. O'Connor","doi":"10.1016/j.ijpddr.2025.100591","DOIUrl":"10.1016/j.ijpddr.2025.100591","url":null,"abstract":"<div><div>Toxoplasmosis is a worldwide parasitic disease caused by the apicomplexan <em>Toxoplasma gondii</em>. Severe neurological illness occurs in immunosuppressed patients, and congenital disorders can follow transplacental primo infection during pregnancy. New effective antiparasitic drugs are needed since chronic cystic stages are resistant to current available treatments, and some of the congenital infections are unresponsive to available therapeutics. Tartrolon E (trtE) is a marine secondary metabolite that has high selectivity against human and animal apicomplexan parasites including <em>T. gondii</em>, <em>Cryptosporidium parvum</em> and <em>Plasmodium falciparum</em>. We evaluated the effect of the compound on extracellular tachyzoite viability, morphology, membrane permeability and its ability to block host cell attachment and/or invasion. While 80 % of <em>T. gondii</em> infective capacity is blocked after only 30 min of compound treatment, parasite viability, morphology, membrane integrity and host cell attachment were unaffected until after 4 h of treatment. These effects were irreversible when parasites were allowed to infect host cells after trtE treatment. Drug exposure for more than 4 h significantly affected tachyzoite survival and altered parasite morphology. The mechanism of action of trtE is still unknown but includes blocking parasite invasion processes. Further research is needed to determine the molecular target of trtE to further progress the compound as an antiparasitic candidate.</div></div>","PeriodicalId":13775,"journal":{"name":"International Journal for Parasitology: Drugs and Drug Resistance","volume":"28 ","pages":"Article 100591"},"PeriodicalIF":4.1,"publicationDate":"2025-04-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143830062","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-03-26DOI: 10.1016/j.ijpddr.2025.100589
Dave Leathwick, Peter Green, Charlotte Bouchet, Alex Chambers, Tania Waghorn, Christian Sauermann
In the faecal egg count reduction test, visual identification of larvae cultured from faeces enables the egg counts to be apportioned to species/genera, resulting in a more accurate test. However, morphology cannot reliably differentiate some species meaning that, in some cases, efficacy can only be estimated at the genus or species-complex level. We investigated the benefits of identifying larvae to species using DNA to determine how often this would alter the diagnosis of resistance and whether increasing the number of larvae identified would alter the repeatability of an efficacy estimate.
Data on faecal nematode egg counts and the corresponding larval species mixes were acquired from tests conducted on commercial sheep farms. The proportion of each species present in faecal culture was determined using DNA. Efficacy was then compared for individual species and for those genera/species complexes which cannot reliably be differentiated visually. The proportion of each species present was subsequently resampled 10,000 times (repeated random sampling) and efficacy recalculated to produce the median efficacy, along with the 5 % and 95 % simulation percentiles. Subsequently, the number of larvae sampled to determine the species mix in each sample was varied from 50 to 6400 and the process repeated.
Of 152 comparisons of efficacy, 25 % of cases where genus-level identification resulted in a finding of ‘susceptible’ for that category, species-level identification returned at least one diagnosis of ‘resistant’ i.e., genus-level identification resulted in a 25 % false negative diagnosis.
When the number of larvae sampled for species identification was low (<400) variation in efficacy estimates was high, however, as sample size increased the confidence interval around the efficacy estimate decreased.
The results indicate that identifying large numbers of larvae to species using DNA has the potential to increase the accuracy and confidence in efficacy estimates achieved using the faecal egg count reduction test.
{"title":"The faecal egg count reduction test: Will identification of larvae to species improve its utility?","authors":"Dave Leathwick, Peter Green, Charlotte Bouchet, Alex Chambers, Tania Waghorn, Christian Sauermann","doi":"10.1016/j.ijpddr.2025.100589","DOIUrl":"10.1016/j.ijpddr.2025.100589","url":null,"abstract":"<div><div>In the faecal egg count reduction test, visual identification of larvae cultured from faeces enables the egg counts to be apportioned to species/genera, resulting in a more accurate test. However, morphology cannot reliably differentiate some species meaning that, in some cases, efficacy can only be estimated at the genus or species-complex level. We investigated the benefits of identifying larvae to species using DNA to determine how often this would alter the diagnosis of resistance and whether increasing the number of larvae identified would alter the repeatability of an efficacy estimate.</div><div>Data on faecal nematode egg counts and the corresponding larval species mixes were acquired from tests conducted on commercial sheep farms. The proportion of each species present in faecal culture was determined using DNA. Efficacy was then compared for individual species and for those genera/species complexes which cannot reliably be differentiated visually. The proportion of each species present was subsequently resampled 10,000 times (repeated random sampling) and efficacy recalculated to produce the median efficacy, along with the 5 % and 95 % simulation percentiles. Subsequently, the number of larvae sampled to determine the species mix in each sample was varied from 50 to 6400 and the process repeated.</div><div>Of 152 comparisons of efficacy, 25 % of cases where genus-level identification resulted in a finding of ‘susceptible’ for that category, species-level identification returned at least one diagnosis of ‘resistant’ i.e., genus-level identification resulted in a 25 % false negative diagnosis.</div><div>When the number of larvae sampled for species identification was low (<400) variation in efficacy estimates was high, however, as sample size increased the confidence interval around the efficacy estimate decreased.</div><div>The results indicate that identifying large numbers of larvae to species using DNA has the potential to increase the accuracy and confidence in efficacy estimates achieved using the faecal egg count reduction test.</div></div>","PeriodicalId":13775,"journal":{"name":"International Journal for Parasitology: Drugs and Drug Resistance","volume":"27 ","pages":"Article 100589"},"PeriodicalIF":4.1,"publicationDate":"2025-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143726167","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-03-20DOI: 10.1016/j.ijpddr.2025.100588
Zachary Caterer , Rachel V. Horejsi , Carly Weber , Blake Mathisen , Chase N. Nelson , Maggie Bagatta , Ireland Coughlin , Megan Wettstein , Ankit Kulshrestha , Hui Siang Benjamin Lee , Leonardo R. Nunn , Mostafa Zamanian , Nicolas J. Wheeler
Image-based phenotypic screening is a fundamental technique used to better understand the basic biology of helminths and advance discovery of new anthelmintics. Miniaturization of screening platforms and automated microscopy have led to a surge in imaging data and necessitated software to organize and analyze these data. Traditionally, these analyses are performed remotely on high-performance computers, often requiring an understanding of a command line interface (CLI) and the ability to write scripts to control the software or job scheduler. Requiring access to specialized computing equipment and advanced computational skills raises the barrier to entry for these sorts of studies. The development of efficient, performant computer and graphical processing units for personal computers and cheaper imaging solutions has made the requirement of remote servers superfluous for many small to medium-scale screens, but most analytical software still require interaction with a CLI. To democratize the analysis of image-based phenotypic screens, we have developed a graphical user interface (GUI) for wrmXpress, a tool that integrates many popular computational pipelines for analyzing imaging data of parasitic and free-living worms. The GUI operates on any personal computer using the operating system's native web browser, allowing users to configure and run analyses using a point-and-click approach. Containerization of the application eliminates the need to install specialized programming libraries and dependencies, further increasing the ease of use. GUI development required a substantial reorganization of the wrmXpress backend codebase, which allowed for the addition of a new pipeline for high-resolution tracking of worm behavior, and we demonstrate its functionality by showing that praziquantel modulates the behavior of Schistosoma mansoni miracidia. These advances make cutting-edge analyses of image-based phenotyping of worms more equitable and accessible.
{"title":"A graphical user interface for wrmXpress 2.0 streamlines helminth phenotypic screening","authors":"Zachary Caterer , Rachel V. Horejsi , Carly Weber , Blake Mathisen , Chase N. Nelson , Maggie Bagatta , Ireland Coughlin , Megan Wettstein , Ankit Kulshrestha , Hui Siang Benjamin Lee , Leonardo R. Nunn , Mostafa Zamanian , Nicolas J. Wheeler","doi":"10.1016/j.ijpddr.2025.100588","DOIUrl":"10.1016/j.ijpddr.2025.100588","url":null,"abstract":"<div><div>Image-based phenotypic screening is a fundamental technique used to better understand the basic biology of helminths and advance discovery of new anthelmintics. Miniaturization of screening platforms and automated microscopy have led to a surge in imaging data and necessitated software to organize and analyze these data. Traditionally, these analyses are performed remotely on high-performance computers, often requiring an understanding of a command line interface (CLI) and the ability to write scripts to control the software or job scheduler. Requiring access to specialized computing equipment and advanced computational skills raises the barrier to entry for these sorts of studies. The development of efficient, performant computer and graphical processing units for personal computers and cheaper imaging solutions has made the requirement of remote servers superfluous for many small to medium-scale screens, but most analytical software still require interaction with a CLI. To democratize the analysis of image-based phenotypic screens, we have developed a graphical user interface (GUI) for wrmXpress, a tool that integrates many popular computational pipelines for analyzing imaging data of parasitic and free-living worms. The GUI operates on any personal computer using the operating system's native web browser, allowing users to configure and run analyses using a point-and-click approach. Containerization of the application eliminates the need to install specialized programming libraries and dependencies, further increasing the ease of use. GUI development required a substantial reorganization of the wrmXpress backend codebase, which allowed for the addition of a new pipeline for high-resolution tracking of worm behavior, and we demonstrate its functionality by showing that praziquantel modulates the behavior of <em>Schistosoma mansoni</em> miracidia. These advances make cutting-edge analyses of image-based phenotyping of worms more equitable and accessible.</div></div>","PeriodicalId":13775,"journal":{"name":"International Journal for Parasitology: Drugs and Drug Resistance","volume":"27 ","pages":"Article 100588"},"PeriodicalIF":4.1,"publicationDate":"2025-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143678955","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-26DOI: 10.1016/j.ijpddr.2025.100587
Autumn Collins , Sierra Varley , Tobias Clark , Nathan Chubb , Sean Forrester , Jean-Paul Desaulniers
The acetylcholine-gated chloride channel (ACC) family in parasitic nematodes represents a promising target for anthelmintic drug development. Levamisole, a widely known and utilized cholinergic agonist, has been used for decades to address many types of parasitic infections by targeting nematode nicotinic acetylcholine receptors (nAChRs) in nematodes. In this study, we report the synthesis and pharmacological evaluation of eight levamisole derivatives, five of which are novel, on the H. contortus ACC-2 receptor. This includes a CF3-derivatized compound we have identified as compound 6 whose structure contains levamisole as a backbone with the addition of a 2-trifluoromethyl benzyl group. Electrophysiological assays revealed that compound 6 exhibited a five-fold increase in sensitivity (EC50 20 μM) compared to levamisole (EC50 100 μM), our parent compound, with an EC50 comparable to that of acetylcholine (20 μM). Investigation of the in silico docking of compound 6 with H. contortus ACC-2 suggest that it interacts uniquely within the H. contortus ACC-2 binding pocket, which may contribute to its increased receptor sensitivity. These findings highlight the potential of structural modifications containing an electron-withdrawing group at the 2-position which can significantly enhance activity at the H. contortus ACC-2 receptor. This opens many avenues for the development of more effective treatments against parasitic nematodes, in an environment with increasing resistance.
{"title":"Increased activity of CF3-derivatized levamisole at the ACC-2 receptor from the parasitic nematode Haemonchus contortus","authors":"Autumn Collins , Sierra Varley , Tobias Clark , Nathan Chubb , Sean Forrester , Jean-Paul Desaulniers","doi":"10.1016/j.ijpddr.2025.100587","DOIUrl":"10.1016/j.ijpddr.2025.100587","url":null,"abstract":"<div><div>The acetylcholine-gated chloride channel (ACC) family in parasitic nematodes represents a promising target for anthelmintic drug development. Levamisole, a widely known and utilized cholinergic agonist, has been used for decades to address many types of parasitic infections by targeting nematode nicotinic acetylcholine receptors (nAChRs) in nematodes. In this study, we report the synthesis and pharmacological evaluation of eight levamisole derivatives, five of which are novel, on the <em>H. contortus</em> ACC-2 receptor. This includes a CF<sub>3</sub>-derivatized compound we have identified as compound <strong>6</strong> whose structure contains levamisole as a backbone with the addition of a 2-trifluoromethyl benzyl group. Electrophysiological assays revealed that compound <strong>6</strong> exhibited a five-fold increase in sensitivity (EC<sub>50</sub> 20 μM) compared to levamisole (EC<sub>50</sub> 100 μM), our parent compound, with an EC<sub>50</sub> comparable to that of acetylcholine (20 μM). Investigation of the <em>in silico</em> docking of compound <strong>6</strong> with <em>H. contortus</em> ACC-2 suggest that it interacts uniquely within the <em>H. contortus</em> ACC-2 binding pocket, which may contribute to its increased receptor sensitivity. These findings highlight the potential of structural modifications containing an electron-withdrawing group at the 2-position which can significantly enhance activity at the <em>H. contortus</em> ACC-2 receptor. This opens many avenues for the development of more effective treatments against parasitic nematodes, in an environment with increasing resistance.</div></div>","PeriodicalId":13775,"journal":{"name":"International Journal for Parasitology: Drugs and Drug Resistance","volume":"27 ","pages":"Article 100587"},"PeriodicalIF":4.1,"publicationDate":"2025-02-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143562769","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-21DOI: 10.1016/j.ijpddr.2025.100586
Chun-Hsien Chen , Jian-Ming Huang , Yu-Jen Wang , Chih-Ming Tsai , Wei-Chen Lin
This review examines the advancements in antimicrobial drug discovery with in vitro assays for Acanthamoeba, highlighting the efficacy of current topical antimicrobial agents. In recent decades, the treatment and diagnosis of Acanthamoeba keratitis (AK) have presented clinical challenges. Clinicians often rely on clinical judgment, risk factors, and patient travel history to guide initial treatment decisions. The clinical presentation of AK frequently coincides with bacterial and fungal keratitis, leading to delays in diagnostic confirmation. This review compiles a list of commonly used antimicrobial agents that may be useful in controlling and preventing Acanthamoeba and other microbial infections during the diagnostic waiting period. Due to their unique life cycle, consisting of both trophozoite and cyst stages, amoebae exhibit resistance to various clinical drugs. Current research efforts are focused on identifying alternative and effective treatment options. Despite the ongoing characterization of various cytocidal agents from natural and synthetic sources, chlorhexidine gluconate (CHG) and polyhexamethylene biguanide (PHMB) have emerged as the most effective therapies for AK. Drawing from previous studies, we catalog several commonly used antimicrobial agents that may enhance the efficacy of PHMB and CHG while also preventing other microbial infections. These alternative agents present promising options for treating AK cases. This review evaluates progress in anti-amoebic drug discovery, focusing on antibiotics and cataloging their activity at different stages of Acanthamoeba.
{"title":"Recent in vitro advances in the ocular antimicrobial agents against Acanthamoeba","authors":"Chun-Hsien Chen , Jian-Ming Huang , Yu-Jen Wang , Chih-Ming Tsai , Wei-Chen Lin","doi":"10.1016/j.ijpddr.2025.100586","DOIUrl":"10.1016/j.ijpddr.2025.100586","url":null,"abstract":"<div><div>This review examines the advancements in antimicrobial drug discovery with in vitro assays for <em>Acanthamoeba</em>, highlighting the efficacy of current topical antimicrobial agents. In recent decades, the treatment and diagnosis of Acanthamoeba keratitis (AK) have presented clinical challenges. Clinicians often rely on clinical judgment, risk factors, and patient travel history to guide initial treatment decisions. The clinical presentation of AK frequently coincides with bacterial and fungal keratitis, leading to delays in diagnostic confirmation. This review compiles a list of commonly used antimicrobial agents that may be useful in controlling and preventing <em>Acanthamoeba</em> and other microbial infections during the diagnostic waiting period. Due to their unique life cycle, consisting of both trophozoite and cyst stages, amoebae exhibit resistance to various clinical drugs. Current research efforts are focused on identifying alternative and effective treatment options. Despite the ongoing characterization of various cytocidal agents from natural and synthetic sources, chlorhexidine gluconate (CHG) and polyhexamethylene biguanide (PHMB) have emerged as the most effective therapies for AK. Drawing from previous studies, we catalog several commonly used antimicrobial agents that may enhance the efficacy of PHMB and CHG while also preventing other microbial infections. These alternative agents present promising options for treating AK cases. This review evaluates progress in anti-amoebic drug discovery, focusing on antibiotics and cataloging their activity at different stages of <em>Acanthamoeba</em>.</div></div>","PeriodicalId":13775,"journal":{"name":"International Journal for Parasitology: Drugs and Drug Resistance","volume":"27 ","pages":"Article 100586"},"PeriodicalIF":4.1,"publicationDate":"2025-02-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143562765","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-01DOI: 10.1016/j.ijpddr.2025.100585
Kateryna Starynets , Ana Paunkov , Anja Wagner , Klaus Kratochwill , Christian Klotz , David Leitsch
The microaerophilic/anaerobic protist Giardia lamblia is a world-wide occurring parasite of the human small intestine. It causes giardiasis which manifests as diarrhoea accompanied by other sequelae. Giardiasis is most commonly treated with either the 5-nitroimidazole metronidazole or the benzimidazole albendazole. Unfortunately, the number of refractory cases is increasing, which is probably caused, at least in part, by drug resistance. However, most attempts to isolate metronidazole-resistant G. lamblia strains from patients have failed so far because the parasites were not resistant when tested in vitro.
We hypothesized that this failure might be caused by drug assay conditions which are standardly anaerobic, and performed metronidazole susceptibility testing with two well studied strains, i.e. WB C6 and BRIS/87/HEPU/713 (strain 713) under microaerophilic conditions. Indeed, 713 proved to be less susceptible to metronidazole under microaerophilic conditions as compared to anaerobic conditions, and residual growth was even noted at concentrations of metronidazole similar to those in the serum of treated patients (i.e. about 100 μM). Further experiments showed that 713 also grows much faster under microaerobic conditions than WB C6. Reduced susceptibility to metronidazole under microaerobic conditions was also observed in a clinical isolate from a refractory giardiasis case.
Two-dimensional gel electrophoresis showed that microaerobic growth was accompanied by the upregulation of superoxide reductase, a pyridoxamine 5′-phosphate oxidase putative domain-containing protein, and a TlpA-like protein in 713 but not in WB C6. All three proteins are known, or can be predicted to have antioxidant functions. Indeed, overexpression of pyridoxamine 5′-phosphate oxidase in WB C6 from a plasmid carrying the respective gene behind the arginine deiminase promoter significantly improved growth of the transfected cell line under microaerobic conditions. Moreover, similarly overexpressed superoxide reductase conferred significant protection against metronidazole.
Our results suggest that oxygen concentrations can affect the outcomes of metronidazole treatment against G. lamblia.
{"title":"Culturing of Giardia lamblia under microaerobic conditions can impact metronidazole susceptibility by inducing increased expression of antioxidant enzymes","authors":"Kateryna Starynets , Ana Paunkov , Anja Wagner , Klaus Kratochwill , Christian Klotz , David Leitsch","doi":"10.1016/j.ijpddr.2025.100585","DOIUrl":"10.1016/j.ijpddr.2025.100585","url":null,"abstract":"<div><div>The microaerophilic/anaerobic protist <em>Giardia lamblia</em> is a world-wide occurring parasite of the human small intestine. It causes giardiasis which manifests as diarrhoea accompanied by other sequelae. Giardiasis is most commonly treated with either the 5-nitroimidazole metronidazole or the benzimidazole albendazole. Unfortunately, the number of refractory cases is increasing, which is probably caused, at least in part, by drug resistance. However, most attempts to isolate metronidazole-resistant <em>G</em>. <em>lamblia</em> strains from patients have failed so far because the parasites were not resistant when tested <em>in vitro</em>.</div><div>We hypothesized that this failure might be caused by drug assay conditions which are standardly anaerobic, and performed metronidazole susceptibility testing with two well studied strains, i.e. WB C6 and BRIS/87/HEPU/713 (strain 713) under microaerophilic conditions. Indeed, 713 proved to be less susceptible to metronidazole under microaerophilic conditions as compared to anaerobic conditions, and residual growth was even noted at concentrations of metronidazole similar to those in the serum of treated patients (i.e. about 100 μM). Further experiments showed that 713 also grows much faster under microaerobic conditions than WB C6. Reduced susceptibility to metronidazole under microaerobic conditions was also observed in a clinical isolate from a refractory giardiasis case.</div><div>Two-dimensional gel electrophoresis showed that microaerobic growth was accompanied by the upregulation of superoxide reductase, a pyridoxamine 5′-phosphate oxidase putative domain-containing protein, and a TlpA-like protein in 713 but not in WB C6. All three proteins are known, or can be predicted to have antioxidant functions. Indeed, overexpression of pyridoxamine 5′-phosphate oxidase in WB C6 from a plasmid carrying the respective gene behind the arginine deiminase promoter significantly improved growth of the transfected cell line under microaerobic conditions. Moreover, similarly overexpressed superoxide reductase conferred significant protection against metronidazole.</div><div>Our results suggest that oxygen concentrations can affect the outcomes of metronidazole treatment against <em>G</em>. <em>lamblia</em>.</div></div>","PeriodicalId":13775,"journal":{"name":"International Journal for Parasitology: Drugs and Drug Resistance","volume":"27 ","pages":"Article 100585"},"PeriodicalIF":4.1,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143179624","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
扫码关注我们
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号