Anthelmintic Research Articles

Glucose transporters and sodium glucose co-transporters cooperatively import glucose into energy-demanding organs in carcinogenic liver fluke Clonorchis sinensis.

The liver fluke Clonorchis sinensis imports large amounts of glucose to generate energy and metabolic intermediates through glycolysis. We hypothesized that C. sinensis absorbs glucose through glucose transporters and identified four subtypes of glucose transporter (CsGTP) and one sodium glucose co-transporter (CsSGLT) in C. sinensis. Expressed sequence tags encoding CsGTPs were retrieved from the C. sinensis transcriptome database, and their full-length cDNA sequences were obtained by rapid amplification of cDNA ends (RACE). The tissue distribution of glucose transporters in C. sinensis adults was determined using immunohistochemical staining. Developmental expression was measured using RT-qPCR. The transport and distribution of glucose into living C. sinensis were monitored using confocal microscopy. Membrane topology and key functional residues of CsGTPs were homologous to their counterparts in animals and humans. CsGTP1, 2, and 4 were transcribed 2.4-5.5 times higher in the adults than metacercariae, while CsGTP3 was transcribed 2.1 times higher in the metacercariae than adults. CsSGLT transcription was 163.6 times higher in adults than in metacercariae. In adults, CsSGLT was most abundant in the tegument; CsGTP3 and CsSGLT were localized in the vitelline gland, uterine wall, eggs, mesenchymal tissue, and testes; CsGTP4 was found in sperm and mesenchymal tissue; and CsGTP1 was mainly in the sperm and testes. In C. sinensis adults, exogenous glucose is imported in a short time and is present mainly in the middle and posterior body, in which the somatic and reproductive organs are located. Of the exogenous glucose, 53.6% was imported through CsSGLT and 46.4% through CsGTPs. Exogenous glucose import was effectively inhibited by cytochalasin B and phlorizin. We propose that CsSGLT cooperates with CsGTPs to import exogenous glucose from the environmental bile, transport glucose across mesenchymal tissue cells, and finally supply energy-demanding organs in C. sinensis adults. Studies on glucose transporters may pave the way for the development of new anthelmintic drugs.

Helminthic Infection and Anthelmintic Drugs: Synthetic and Herbal Approaches

Helminthic Infection and Anthelmintic Drugs: Synthetic and Herbal Approaches

The effect of anthelmintic treatment and efficacy on the faecal microbiota of healthy adult horses

Changes to the faecal microbiota of horses associated with administration of anthelmintic drugs is poorly defined. This study included horses with cyathostomin infection where susceptibility and resistance to oxfendazole and abamectin was known. This study assessed the changes to the faecal microbiota associated with administration of two different anthelmintics in this population. Twenty-four adult horses were included. Faecal egg counts were performed on all horses prior to random allocation into abamectin (n=8), oxfendazole (n=8) or Control groups (n=8) and at Day 14 post treatment. Faecal samples were collected for microbiota analysis prior to anthelmintic administration and on Day 3 and Day 14. From each faecal sample, DNA was extracted prior to PCR amplification, next generation sequencing and analysis using QIIME2. Anthelmintic treatment was associated with changes in alpha diversity (p <0.05), with increased evenness and diversity at Day 14 and increased richness at Day 3 within the abamectin group. Differences in relative abundance of bacteria at the phyla, family and genus taxonomic levels occurred after treatment; indicating that the microbiota was altered with anthelmintic administration. The results support that anthelmintic administration and removal of cyathostomins from the large intestine of horses is associated with changes in the faecal microbiota. The results suggest that removal of cyathostomins is associated with greater differences in microbiota, compared to anthelmintic drug administration that is ineffective in reducing cyathostomin infection. Cyathostomin removal was supported by adequate reduction of faecal egg counts, determined by faecal egg count reduction testing.

Antihistamines H1 as Potential Anthelmintic Agents against the Zoonotic Parasite Angiostrongylus cantonensis.

Infections caused by parasitic helminths pose significant health concerns for both humans and animals. The limited efficacy of existing drugs underscores the urgent need for novel anthelmintic agents. Given the reported potential of antihistamines against various parasites, including worms, this study conducted a screening of clinically available antihistamines against Angiostrongylus cantonensis-a nematode with widespread implications for vertebrate hosts, including humans. Twenty-one anti-H1 antihistamines were screened against first-stage larvae (L1) of A. cantonensis obtained from the feces of infected rats. Standard anthelmintic drugs ivermectin and albendazole were employed for comparative analysis. The findings revealed four active compounds (promethazine, cinnarizine, desloratadine, and rupatadine), with promethazine demonstrating the highest potency (EC50 = 31.6 μM). Additionally, morphological analysis showed that antihistamines induced significant changes in larvae. To understand the mechanism of action, antimuscarinic activities were reported based on average pK i values for human muscarinic receptor (mAChR) subtypes of the evaluated compounds. Furthermore, an analysis of the physicochemical and pharmacodynamic properties of antihistamines revealed that their anthelmintic activity does not correlate with their activity at H1 receptors. This study marks the first documentation of antihistamines' activity against A. cantonensis, offering a valuable contribution to the quest for novel agents effective against zoonotic helminths.

Veterinary Drug Use Pattern in livestock and Its Public Health Significance, Gondar Town Veterinary Clinic, Northern Ethiopia

A cross-sectional and retrospective study design and simple random sampling strategy was conducted in Gondar town veterinary clinic, northern Ethiopia with the aim of evaluating drug use pattern and show public health significance of irrational veterinary drug use. A total of 2117 drugs were prescribed to 1717 veterinary patients randomly selected from the case registry in this study. The result indicates average number of drugs prescribed per case was 1.23 and the maximum of three drugs was prescribed. The percentages of antimicrobial, anthelmintic, endectocide and other drugs prescribed were 1182 (55.7%), 481 (22.7%), 426 (20.1%), and 28 (1.3%), respectively. All patients were treated without correct laboratory support. As a result, antimicrobials were inappropriately prescribed in 1.4% of viral diseases, 0.2% of external parasitic diseases, 0.8% of internal parasitic diseases, 0.3% of metabolic diseases, 0.1% of fungal diseases, and anthelmintics in 0.1% of bacterial diseases and 0.5% of external parasitic diseases. 2013/2117 (95.1%) the route of administration of the prescribed drugs was not stated. Clinic professionals were 80% animal health assistants and 20% veterinarians. The results reveal problems with correct diagnosis, low level of education of prescribers, few essential drugs, absence of standard veterinary drug lists, and inappropriate drug use; these all principals to public health significance. Therefore, the availability of key essential drugs should improve and available drugs be prescribed with its appropriate doses, routes, and regimens. Veterinarians should be made aware of the irrational use of veterinary drugs and their public health implications.

Unravelling the Effectiveness of Anthelmintic Treatments on Equine Strongyles on Irish Farms.

Over the preceding decades, the widespread dependence on anthelmintic drugs for managing nematodes in grazing equids has given rise to resistance against commonly used anthelmintics in various countries. This study explores the prevalence of anthelmintic resistance across 44 horse farms in Ireland. Anthelmintic efficacy was evaluated through fecal egg count reduction (FECR) tests employing the mini-FLOTAC technique. Resistance to benzimidazoles was identified in 12 out of 14 farms (FECR range: 0.00% to 86.2%). Ivermectin resistance was observed on two farms, one with an FECR of 80.70% and another with an FECR of 96.10% (lower 95% high probability density interval (HPD) <90%, 11.70%). On the remaining six farms, the reduction with ivermectin still exceeded 95%. The reduced efficacy of moxidectin was noted on two farms (FECR = 86.90% and 93.50%) and on a third farm with an FECR of 99.50 and a lower HPD interval < 90% at 24.00%. In summary, these findings emphasize the urgent need for alternative strategies in equine strongyle control that reduce reliance on anthelmintics and prioritize effective management practices on Irish equine farms to hinder the impending development of drug-resistant parasite populations.

Anthelmintic resistance in the canine hookworm,Ancylostoma caninum

Hookworms are one of the most common soil-transmitted helminths and can infect a wide variety of mammals. There are three major anthelmintic drug classes currently available for treatment of hookworms in canines: the benzimidazoles, the tetrahydropyrimidines and the macrocylic lactones. In registration studies for the food and drug administration in the USA, febantel, moxidectin and milbemycin oxime all demonstrated efficacy of &gt;99%. Fenbendazole demonstrated efficacy of &gt;98% and pyrantel demonstrated a somewhat variable efficacy, with a mean across studies of approximately 94%, where more than half of those studies yielded &gt;99%. The only mechanism of resistance currently known for anthelmintics is for the benzimidazole drugs. Detection of drug-resistant Ancylostoma caninum in dogs is a complex issue that deserves greater recognition before more multiple anthelmintic drug-resistant A. caninum infections are reported in an even wider geographic area and dog population distribution. Beyond the concerns for canine health, multiple anthelmintic drug-resistance in canine hookworms could present serious public health concerns because A. caninum is zoonotic.

Investigation of hypoglycemic and antidiarrheal activity on mice model and in vitro anthelmintic study of Macropanax dispermus (Blume) Kuntze (Araliaceae): a promising ethnomedicinal plant

Introduction: Macropanax dispermus is an ethnomedicinal plant, which has been shown to have pharmacological effects against inflammation. Inflammation is also associated with diabetic-related complications. In addition, the indigenous people used this plant to cure numerous digestive system disorders. These ethnomedicinal uses have inspired the current research to focus on determining the biological potential of the crude methanol extract of its leaves (MDML) and stem barks (MDMS) against morbid diseases such as diarrhea, helminthiasis, and diabetes mellitus related to hyperglycemia. This study also identified the functional groups present in MDML which could correlate the presence of different phytochemicals responsible for biological activities. Methods: The hypoglycemic activity and antidiarrheal tests were done on Swiss albino mice through an oral glucose tolerance test (OGTT) and castor-oil-induced diarrhea, respectively. The anthelmintic test was done in vitro on Pheretima posthuma. The functional groups were identified by using Fourier-transform infrared (FTIR) spectroscopy. Results: Here, MDML at 400 mg/kg exhibited a significant (P&lt;0.05) hypoglycemic activity as well as decreased diarrheal drops significantly (P&lt;0.001) when compared to the control group. Additionally, MDMS showed a concentration-dependent significant effect as a prospective anthelmintic agent. The FTIR spectra demonstrated the presence of different functional groups, which might contribute to the presence of flavonoids, saponins, terpenoids, and other related compounds. Conclusion: The present research demonstrated that MDML and MDMS might be effective interventions for treating hyperglycemia, diarrhea, and helminthiasis.

Beyond parasitism: Exploring the microbial profile of Haemonchus contortus and its predilection site (abomasum) in Kashmir Merino sheep

Gastrointestinal helminth infection, particularly by Haemonchus contortus, poses significant challenges to sheep farming worldwide. While anthelmintic drugs have been traditional control measures, the emergence of resistance calls for alternative strategies. Understanding the interaction between parasites, host, and their microbiome is crucial for management of helminth infection. This study intricately explores the interactions between microbial communities in Kashmir Merino sheep infected with H. contortus, to understand the complex interplay between host, parasite, and their microbiome. Sheep abomasal contents and H. contortus were collected from infected and control groups, processed for DNA extraction, and subjected to metagenomic sequencing of the 16 S rRNA gene. Downstream analysis unveils distinct microbial patterns, where Proteobacteria were dominant in H. contortus, while Bacteroidota and Firmicutes prevailed in the sheep abomasum. The revelation of unique genera and shifts in diversity indices underscored helminth-induced disruptions in the host. Beta diversity analysis further showed significant variations in bacterial profiles, providing insights into the intricate host, parasite, and microbiome dynamics. Additionally, this study elucidated the presence of pathogenic bacteria within H. contortus, accentuating their potential role in exacerbating sheep health issues. This finding underscores the complexity of the host-parasite-microbiome interaction showing helminth-induced microbiome alterations of the host.

Bioanalytical Method Development and Validation of Antihelmintic Drug in Human Plasma

Bioanalytical Method Development and Validation of Antihelmintic Drug in Human Plasma

Levamisole Impairs Vascular Function by Blocking α-Adrenergic Receptors and Reducing NO Bioavailability in Rabbit Renal Artery

Levamisole is an anthelmintic drug restricted to veterinary use but is currently detected as the most widely used cocaine cutting agent in European countries. Levamisole-adulterated cocaine has been linked to acute kidney injury, marked by a decrease in glomerular filtration rate, which involves reduced renal blood flow, but data on the alteration of renovascular response produced by levamisole are scarce. Renal arteries were isolated from healthy rabbits and used for isometric tension recording in organ baths and protein analysis. We provide evidence that depending on its concentration, levamisole modulates renovascular tone by acting as a non-selective α-adrenergic receptor blocker and down-regulates α1-adrenoceptor expression. Furthermore, levamisole impairs the endothelium-dependent relaxation induced by acetylcholine without modifying endothelial nitric oxide synthase (eNOS) expression. However, exposure to superoxide dismutase (SOD) partially prevents the impairment of ACh-induced relaxation by levamisole. This response is consistent with a down-regulation of SOD1 and an up-regulation of NADPH oxidase 4 (Nox4), suggesting that endothelial NO loss is due to increased local oxidative stress. Our findings demonstrate that levamisole can interfere with renal blood flow and the coordinated response to a vasodilator stimulus, which could worsen the deleterious consequences of cocaine use.Graphical EFS electric field stimulation, NA noradrenaline, AR adrenergic receptor, IP3 inositol 1, 4, 5-trisphosphate, cAMP cyclic adenosine monophosphate, mAChR muscarinic acetylcholine receptor, eNOS endothelial nitric oxide synthase, sGC soluble guanylyl cyclase, SOD superoxide dismutase, NOX4 NAPH oxidase 4

Phytochemical Study of the Anthelminthic Potential of Guadeloupean Plant Biodiversity.

Gastrointestinal parasitism is a major health and welfare problem in ruminants. Synthetic chemical anthelmintic drugs have led to the emergence of resistance in gastrointestinal strongyles, inducing the search for alternatives to control the infections that affect ruminants. The objective of this work was to evaluate the anthelmintic potential of plant extracts against Haemonchus contortus Rudolphi. Three plants of the Guadeloupean biodiversity, Momordica charantia L., Carica papaya L. and Sargassum spp., were selected based on their high polyphenolic content and natural abundance. The phytochemistry of plants was explored, a biological assay against the parasite H. contortus was carried out, and several hypotheses about the way of action were proposed by an innovative electrochemical screening method.

GASP: A Pan-Specific Predictor of Family 1 Glycosyltransferase Acceptor Specificity Enabled by a Pipeline for Substrate Feature Generation and Large-Scale Experimental Screening.

Glycosylation represents a major chemical challenge; while it is one of the most common reactions in Nature, conventional chemistry struggles with stereochemistry, regioselectivity, and solubility issues. In contrast, family 1 glycosyltransferase (GT1) enzymes can glycosylate virtually any given nucleophilic group with perfect control over stereochemistry and regioselectivity. However, the appropriate catalyst for a given reaction needs to be identified among the tens of thousands of available sequences. Here, we present the glycosyltransferase acceptor specificity predictor (GASP) model, a data-driven approach to the identification of reactive GT1:acceptor pairs. We trained a random forest-based acceptor predictor on literature data and validated it on independent in-house generated data on 1001 GT1:acceptor pairs, obtaining an AUROC of 0.79 and a balanced accuracy of 72%. The performance was stable even in the case of completely new GT1s and acceptors not present in the training data set, highlighting the pan-specificity of GASP. Moreover, the model is capable of parsing all known GT1 sequences, as well as all chemicals, the latter through a pipeline for the generation of 153 chemical features for a given molecule taking the CID or SMILES as input (freely available at https://github.com/degnbol/GASP). To investigate the power of GASP, the model prediction probability scores were compared to GT1 substrate conversion yields from a newly published data set, with the top 50% of GASP predictions corresponding to reactions with >50% synthetic yields. The model was also tested in two comparative case studies: glycosylation of the antihelminth drug niclosamide and the plant defensive compound DIBOA. In the first study, the model achieved an 83% hit rate, outperforming a hit rate of 53% from a random selection assay. In the second case study, the hit rate of GASP was 50%, and while being lower than the hit rate of 83% using expert-selected enzymes, it provides a reasonable performance for the cases when an expert opinion is unavailable. The hierarchal importance of the generated chemical features was investigated by negative feature selection, revealing properties related to cyclization and atom hybridization status to be the most important characteristics for accurate prediction. Our study provides a GT1:acceptor predictor which can be trained on other data sets enabled by the automated feature generation pipelines. We also release the new in-house generated data set used for testing of GASP to facilitate the future development of GT1 activity predictors and their robust benchmarking.

Vanadium nitride /poly(0-methoxy aniline)- poly(3,4-ethylene dioxythiophene) interpenetrated into nanocomposite for efficient photocatalytic degradation of methylene blue and enhanced electrochemical sensing of mebendazole

Vanadium nitride-poly (0-methoxy aniline)- poly(3,4-ethylene dioxythiophene) (VN-POMA-PEDOT) hybrid was synthesized via ammonolysis and chemical oxidative polymerization technique using VN-POMA-PEDOT/GCE with electrocatalytic activity has two dimensional VN hierarchical porosity with POMA-PEDOT structure created VN-POMA-PEDOT modified GCE working electrode. Donor-acceptor behavior and double-layer growth enable enhanced electrochemical performance and catalytic activity of mebendazole (MBZ). This work investigated the electrochemical sensing conduct of a VN-POMA-PEDOT hybrid composite towards MBZ. The detection limit (DL) and quantification limit (QL) were determined to be 2.192 × 10−9 μM μA−1 and 5.245 × 10−9 M μA−1. Estimation of anti-interference ability, long-term stability, and reproducibility revealed that the prepared VN-POMA-PEDOT electrode is appropriate for the electrochemical sensing finding of MBZ in real analysis, such by way of anti-helminthic drug milk. The VN-POMA-PEDOT achieved 98.9% efficiency in the photocatalytic degradation of methylene blue (MB) within 50 min with degradation rate 8.3 × 10−3 min−1. The suppleness of this method was confirmed by the hybrid morphology VN-POMA-PEDOT, which shows an enormously superior and enhanced photocatalytic presentation of MB.

An automated, high-resolution phenotypic assay for adult Brugia malayi and microfilaria

Brugia malayi are thread-like parasitic worms and one of the etiological agents of Lymphatic filariasis (LF). Existing anthelmintic drugs to treat LF are effective in reducing the larval microfilaria (mf) counts in human bloodstream but are less effective on adult parasites. To test potential drug candidates, we report a multi-parameter phenotypic assay based on tracking the motility of adult B. malayi and mf in vitro. For adult B. malayi, motility is characterized by the centroid velocity, path curvature, angular velocity, eccentricity, extent, and Euler Number. These parameters are evaluated in experiments with three anthelmintic drugs. For B. malayi mf, motility is extracted from the evolving body skeleton to yield positional data and bending angles at 74 key point. We achieved high-fidelity tracking of complex worm postures (self-occlusions, omega turns, body bending, and reversals) while providing a visual representation of pose estimates and behavioral attributes in both space and time scales.

HPLC-ESI-QTOF-MS/MS Analysis of Artemisia cina Leaves Aqueous Extract and Flavonoid Isolation Using High-speed Countercurrent Chromatography

Artemisia cina Berg ex Poljakov, a shrubby plant endemic to the semi-arid areas of Kazakhstan, has as a long history of being used in traditional medicines as an antitumour, antifungal and anthelmintic agent. In a preliminary study carried out on different methodologies to produce aqueous extracts of A. cina it was seen that a simple aqueous extract of A. cina (as in an infusion of the plant) had the strongest anti-SARS-CoV-2 activity. The profiling of this aqueous extract was performed using Liquid Chromatography-Electrospray Ionization-Quadrupole-Time-of-Flight Mass Spectrometry (LC-ESI-Q-TOF-MS) while High-Speed Countercurrent Chromatography (HSCCC) and a Sephadex LH-20 column were utilised to isolate and purify the extract in ethyl acetate obtained from the aqueous extract. Nuclear Magnetic Resonance (NMR) was used for structural elucidation. The flavonoid spinacetin-7-O-glucoside was found to be one of the main compounds present in the A. cina aqueous extract. With the emergence of the COVID-19 pandemic and the frequency in which new COVID strains occur, the demand for drugs with anti-SARS-CoV-2 activity is at an all-time high. Flavonoids and artemisinin identified by LC-MS in this extract are hypothesised to be the key compounds behind the anti-SARS-CoV-2 activity of the aqueous extract, however this needs to be further tested and explored.

Behavioural response to gastrointestinal parasites of yearling dairy calves at pasture

ABSTRACT Aims To investigate the association between gastrointestinal parasites (GIP) and animal behaviour in dairy calves under New Zealand pastoral conditions, using animal-mounted, accelerometer-based sensors. Methods Thirty-six, 5–6-month-old, Friesian-Jersey, heifer calves fitted with animal activity sensors to track behaviour were randomly allocated to one of two treatment groups. Half the animals were challenged with an oral dose of 20,000 larvae of Ostertagia ostertagi and Cooperia oncophera once a week for 3 weeks and half were unchallenged. Five weeks after the last dose, seven infected and nine uninfected animals were treated with an oral anthelmintic (AHC) and data collected for a further week. Accelerometer data were classified into minutes per day eating, ruminating, in moderate–high activity or in low activity. Live weight and faecal egg counts (FEC) were recorded weekly over the study period. All animals co-grazed a newly sown pasture not previously grazed by ruminants and were moved every week to fresh grazing. Treatment status was blinded to those managing the animals which were otherwise treated identically. Results Complete behavioural records were available from 30/36 calves, (13 challenged and 17 unchallenged). Before treatment with AHC, FEC increased in infected and un-treated calves over the study, while uninfected animals maintained a near zero FEC. There was no difference in live weight gain between the two groups over the study period. Bayesian, multinomial regression predicted differences in animal behaviour between infected and uninfected animals that were not treated with AHC over the 7 weeks following initial infection. Parasitised calves not treated with AHC were less active and spent up to 6 (95% highest density interval (HDI) = 1–11) minutes/day less in low level activity and up to 15 (95% HDI = 7–20) minutes/day less in moderate to high level activity. They ruminated up to 9 (95% HDI = 2–15) minutes/day more and ate up to 10 (95% HDI = 2–19) minutes/day more than control calves that were not treated with AHC. The effect of AHC on time spent in each behaviour differed between infected and uninfected calves and increased the coefficient of dispersion of the behavioural data. Conclusions and clinical relevance Small differences in animal behaviour can be measured in calves with GIP. However, to use this to target treatment, further validation studies are required to confirm the accuracy of behavioural classification and understand the complex drivers of animal behaviour in a dynamic and variable pasture-parasite–host environment.

Use of Viscous medium to study anthelmintic drug action in Caenorhabditis elegans

Caenorhabditis elegans is an appealing tool for experimental evolution and for working with antiparasitic drugs, from understanding the molecular mechanisms of drug action and resistance to uncover new drug targets. We present a new methodology for studying the impact of antiparasitic drugs in C. elegans. Viscous medium was initially designed for C. elegans maintenance during long-term evolution experiments. Viscous medium provides a less structured environment than the standard nematode growth media agar, yet the bacteria food source remains suspended. Further, the Viscous medium offers the worm population enough support to move freely, mate, and reproduce at a rate comparable to standard agar cultures. Here, the Viscous medium was adapted for use in antiparasitic research. We observed a similar sensitivity of C. elegans to anthelmintic drugs as in standard liquid media and statistical difference to the standard agar media through a larval development assay. Using Viscous medium in C. elegans studies will considerably improve antiparasitic resistance research, and this medium could be used in studies aimed at understanding long-term multigenerational drug activity.

“Niclosamide: A potential antipruritic agent by modulating serotonin pathway through metabotropic glutamate receptors (mGluRs)”

Pruritus is an uncomfortable sensation induced by various pruritogens, including serotonin. Serotonin, acting as an inflammatory mediator, can activate a histamine-independent pathway. Consequently, many anti-pruritus medications, such as antihistamines, are not effective in adequately relieving patient symptoms. Niclosamide, an anthelmintic drug, has recently demonstrated an affinity for Metabotropic glutamate receptors (mGluRs). mGluRs are a group of receptors activated by glutamate, and they are involved in regulating neuronal excitability. In this study, we utilized mouse models of serotonergic itch and administered different doses of Niclosamide to examine the expression of mGluR1, mGluR5, and 5-HT2. The administration of 5 mg/kg Niclosamide successfully suppressed pruritus in the mice. Additionally, the levels of mGluR1, mGluR5, 5-HT2, and TRPV1 were significantly reduced. These findings suggest that Niclosamide holds promise as a potential antipruritic drug.

Anthelmintic activity and pathophysiological effect of anthelmintic drugs against carcinogenic liver fluke, Opisthorchis viverrini.

Human liver fluke, Opisthorchis viverrini poses a significant risk for development of cholangiocarcinoma (CCA) in Thailand, primarily attributed to consumption of undercooked cyprinoid fishes. The current use of anthelmintic drug treatment such as praziquantel (PZQ), as the main therapeutic agent against O. viverrini. There is a need to explore the efficacy of alternative anthelmintic drugs for O. viverrini treatment. This study aimed to assess the efficacy of anthelmintic drugs, which are commonly use in endemic areas of Southeast Asian countries; PZQ, albendazole (AL), niclosamide (NI), and mebendazole (ME) at concentrations of 600, 400, 500, and 500 mg/ml. The study included a negative and positive control group treated with roswell park memorial institute (RPMI) and PZQ. Reactive oxygen species (ROS) levels, indicative of oxidative stress, were quantified using 2',7'-dichlorofluorescein diacetate staining. Morphological changes were observed using scanning electron microscopy. Furthermore, motility assessments were conducted at various time points (0, 5, 30 minutes, 1, 3, 6, 12, and 24 hours), calculating relative motility (RM) and survival index (SI). The results revealed a significant increase of ROS levels with the intensity and corrected total worm fluorescence (CTWF) mostly observed in order of PZQ, followed by NI, ME, and AL, respectively. Morphological damage was presented the tegumental swelling, papillae changes, and disruption of microvilli (Mv), particularly in the group treated with the most effective anthelmintics PZQ, NI, ME, and AL, while negative control group did not exhibit such alterations. Also, the most efficacy for suppressing the motility of adult worms were displayed in PZQ treatment group, followed by NI, ME, and AL, respectively. Overall, first novel findings suggest that apart from NI, ME, and AL demonstrate potential as alternative therapeutic options for O. viverrini infection. Furthermore, animal model is needed to investigate the efficacy of NI, ME, and AL compare with standard treatment.