Anthelmintic Action Research Articles

On the issue of experimental modeling of invasions and prospects for the use of immunostimulants in nematodes

Currently, the search for ways to overcome drug resistance to anthelmintics is relevant. This problem is solved by limiting the duration of use of drugs with the same mechanism of action and combination of anthelmintics. The studies presented in the article have shown that the use of immunotropic drugs is also very promising. Forty white mice and 20 white rats were used for experimental modeling of Trichinella spiralis and Trichinella nativa invasions. Changes in lymphoid tissue associated in the intestine, in the spleen, as well as in the peripheral blood of animals and birds during inoculation of the above species were studied. Further, the effect of drugs with immunotropic activity was studied on proven models of Trichinella spiralis invasions, for which 30 white mice (20.5±2.5 g) were used, which were randomized and divided into groups. The evaluation of the protective properties of immunotropic drugs in trichinosis was carried out on laboratory models to determine the survival rate in studies after euthanasia of all mice and counting the number of Trichinella larvae. Both experiments were conducted in accordance with the principles of the Helsinki Declaration. After removing laboratory animals from the experiment (in accordance with the principles of the above declaration), macro- and microanatomic studies and sampling of histological material (sets of intestines and spleens) were carried out for qualitative and quantitative assessment. The above is confirmed by the fact that parasitological studies have established the high effectiveness of polyoxidonium in relation to the survival of Trichinella larvae, which was accompanied by an increase in the area of lymphoid tissue associated with the intestine and the white pulp of the spleen, in which lymphoid follicles grew. Thus, polyoxidonium is an effective drug that can be used in the complex treatment of nematodes as an immunotropic agent.

Influence of fecal fermentation on the anthelmintic activity of proanthocyanidins and ellagitannins against human intestinal nematodes and Caenorhabditis elegans.

Some tannin-rich plants such as Combretum mucronatum and Phyllanthus urinaria are widely used in Africa for the control of parasitic nematodes in both humans and livestock. Tannins have been recognized as an alternative source of anthelmintic therapies, and hence, recent studies have focused on both the hydrolyzable and condensed tannins. These groups of compounds, however, have poor oral bioavailability and are metabolized by gut microbiota into lower molecular weight compounds. The role of these metabolites in the anthelmintic activities of tannins has not been explored yet. This study investigated the effects of fecal metabolism on the anthelmintic potential of procyanidin C1 (PC1) and geraniin and the tannin-enriched extracts of C. mucronatum (CML) and P. urinaria (PUH), which contain these compounds, respectively. Metabolites were formed by anaerobic fermentation of the test compounds and extracts in a fresh human fecal suspension for 0 h, 4h, and 24h. Lyophilized samples were tested in vitro against hookworm larvae and whipworm (Trichuris trichiura) larvae obtained from naturally infected human populations in Pru West District, Bono East Region, Ghana, and against the wildtype strain of Caenorhabditis elegans (L4). Both extracts and compounds in the undegraded state exhibited concentration-dependent inhibition of the three nematodes. Their activity, however, significantly decreased upon fecal metabolism. Without fermentation, the proanthocyanidin-rich CML extract was lethal against hookworm L3 (LC50 = 343.5μg/mL, 95% confidence interval (CI) = 267.5-445.4), T. trichiura L1 (LC50 = 230.1μg/mL, CI = 198.9-271.2), and C. elegans (LC50 = 1468.1μg/mL, CI = 990.3-1946.5). PUH, from which the ellagitannin geraniin was isolated, exhibited anthelmintic effects in the unfermented form with LC50 of 300.8μg/mL (CI = 245.1-374.8) against hookworm L3 and LC50 of 331.6μg/mL (CI = 290.3-382.5) against T. trichiura L1, but it showed no significant activity against C. elegans L4 larvae at the tested concentrations. Similarly, both compounds, procyanidin C1 and geraniin, lost their activity when metabolized in fecal matter. The activity of geraniin at a concentration of 170μg/mL against C. elegans significantly declined from 30.4% ± 1.8% to 14.5% ± 1.5% when metabolized for 4h, whereas that of PC1 decreased from 32.4% ± 2.3% to 8.9% ± 0.9% with similar treatment. There was no significant difference between the anthelmintic actions of metabolites from the structurally different tannin groups. The outcome of this study revealed that the intact bulky structure of tannins (hydrolyzable or condensed) may be required for their anthelmintic action. The fermented products from the gut may not directly contribute toward the inhibition of the larvae of soil-transmitted helminths.

A Systematic Review of the Impact of Benzimidazole-based Anthelmintics on Lung Cancer in Animal Models.

Emerging studies have reported the potential anticancer activity of FDA-approved benzimidazole-based anthelmintics against lung cancer. Therefore, the current systematic review aimed to explore the anticancer activity of benzimidazole-based anthelmintics in lung cancer animal models. The databases including Pubmed, ScienceDirect, and Google Scholar were searched till April 2024 for the animal studies evaluating the anticancer activity of benzimidazole-based anthelmintics against lung cancer. The relevant data was extracted in the prepared format in Microsoft Excel. Systematic Review Center for Laboratory Animal Experimentation (SYRCLE) risk of bias (RoB) was used to assess the quality of included studies. The protocol for this study has been registered in PROSPERO (Registration number: CRD42022352141). Initially, we obtained 4150 articles, and finally eight articles were included in the current study. The information in the included studies was a bit diversified including different benzimidazole-based anthelmintics, dosage, route of administration, and duration of experiments. However, all studies reported that exposure to benzimidazole-based anthelmintics decreased tumor size and tumor volume in animal models of lung cancer. In conclusion, benzimidazole-based anthelmintics have the potential to treat lung cancer. However, more controlled and thorough preclinical studies are required to evaluate its efficacy, safety, and mechanism of anticancer activities.

Antioxidant, Anti-inflammatory and Anthelminthic Activities of Traditionally used Ocimum basilicum L., and Ocimum tenuiflorum L. Extract Loaded Emulsion

The demand for cosmeceuticals is rapidly expanding. Various phytoconstituents are identified from Ocimum basilicum and O. tenuiflorum and have several pharmacological properties. The present study aimed to formulate a novel alternative herbal formulation for topical administration with natural antioxidant properties for the treatment of inflammatory conditions with the destructive capacity of parasitic worms. The egg albumin denaturation technique was used to evaluate the test formulations’ and the standard’s in-vitro anti-inflammatory properties, and the hydrogen peroxide assay was used to measure the extracts’ antioxidant properties. Eudrilus eugeniae was the target of the investigation into in-vitro anthelmintic action. The pH of the cream was 6.8 (average), it was easily spreadable and washable, no gritty particles were found and the dye test confirmed that it was an o/w type of cream. The egg albumin denaturation technique was utilized to evaluate the anti-inflammatory properties. The results cleared those herbal formulations (F1 and F2) and protein denaturation was decreased by diclofenac sodium (Standard) in a dose-dependent manner. Test samples exhibited higher anti-inflammatory activities than the standard with their percentage inhibitions being 41.5 ± 0.5 (F2), 39.1 ± 0.4 mg/mL (F1), and 37.3 ± 0.3 mg/mL (F-Standard), respectively. Antioxidants are compounds that either completely stop or significantly reduce the harm that free unstable radicals. F2 showed a higher concentration-dependent anthelmintic activity than F1 and standard albendazole. Increasing the concentration decreases the paralysis and death time of earthworms. Finally concluded that F2 indicates higher anti-inflammatory and anthelmintic activities than others with high natural antioxidant power.

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.

Addressing Toxicity Concerns: State-of-the-Art Synthesis Methods and Emerging Multifaceted Applications of Silver Nanoparticles

Abstract: Silver nanoparticles (AgNPs), typically ranging in size from 10 to 1000 nm, are synthesize through physical, chemical, or biological methods. The biological approach, which often uses plant extracts, microbes, or fungi, is recommended for its eco-friendliness and lower dangers, while physical and chemical procedures can be costly and potentially dangerous. AgNPs are widely used in treatments and diagnostics in the medical field. Their exceptional antibacterial, antifungal, antiviral, anticancer, antiangiogenic, leishmanicidal, anti-inflammatory, antidiabetic, antioxidant, and anthelmintic action are just a few of their special qualities. However, AgNPs do have certain disadvantages due to worries about their nanotoxicity. This thorough study covers a wide range of AgNP topics, including their synthesis, properties, factors affecting particle size, and modes of action. Additionally, it covers their uses in the medical field, any potential toxicity issues, and the difficulties associated with their application.

Phytochemical Study, Evaluation of Antihelminthic Potential and In silico Screening for Breast Cancer Using Mimosa pudica Linn.

Breast Cancer is a major health concern in India, where it is highly lethal for women because of unchecked cell proliferation and metastasis, which are frequently caused by interactions between the Estrogen Receptor Alpha and other receptors. Even though they work well, synthetic Anthelmintics come with dangers to the health of both humans and animals. Therefore, creating effective and safe Anthelmintics from plant sources is our main goal. We extracted Mimosa pudica leaves with ethanol using a Soxhlet system, and after concentration, we examined the extract using GC-MS, FTIR, and UV techniques. Estrogen Receptor structure and flavonoid compound databases from PubChem and Protein Data Bank were used for in silico testing against Breast Cancer and anthelmintic action, respectively. Using molecular docking and drug similarity investigations, the effectiveness of natural compounds against Breast Cancer was evaluated.

Hybrids of imidazole with indoline derivatives: Microwave assisted synthesis, molecular docking studies, possible biological activities

A new hybrids of imidazole with Indole derivative 1-methyl-3-((1-(2-methyl-1H-Imidazole.-1yl)-methyl)-1H-indol-5-yl)imino)indolin-2-one(3a-o) was prepared by microwave-assisted technique via alkylation and Schiff's and Mannich base mechanism. The chemical structures of the synthesized products were confirmed by IR, 1HNMR and Mass spectra. The in vitro anticancer activity was evaluated via MTT assay technique, as well as their anthelmintic activity by utilising Indian earth warms. Most of the compounds 1-methyl-3-((1-(2-methyl-1H-Imidazole-1yl)-methyl)-1H-indol-5-yl)imino)indolin-2-one demonstrated greater anthelmintic action than the other compounds. The compounds 1,5-dimethyl-3-((1-(2-methyl-1H-Imidazole-1-yl)-methyl)-1H-indol-5-yl)-imino)-indole-2-one and 1-benzyl-5--chloro-3-((1-((2-methyl-1H--imidazole-1-yl) methyl)-1H-indol-5-yl) imino)indolin-2-one exhibit good anticancer action. Lastly, whenever the interaction pattern and selectivity towards the EGFR activity site have been examined, the molecular docking experiments of novel indole derivatives demonstrated good alignment only with biological findings.

Anti-Opisthorchis felineus effects of artemisinin derivatives: An in vitro study

BackgroundThe drug of choice for the treatment of opisthorchiasis caused by trematodes Opisthorchis viverrini and O. felineus is praziquantel (PZQ), but there is a constant search for new anthelmintics, including those of plant origin. Positive results on the use of artemisinin derivatives against O. viverrini opisthorchiasis have been shown previously, but the effect of these compounds on O. felineus has not been studied. Therefore, here, a comparative analysis of anthelmintic properties of artemisinin derivatives (artesunate [AS], artemether [AM], and dihydroartemisinin [DHA]) was carried out in vitro in relation to PZQ. Experiments were performed on newly excysted metacercariae (NEMs) and adult flukes of O. felineus. ResultsDose- and time-dependent effects of artemisinin derivatives and of PZQ were assessed in terms of motility and mortality of both NEMs and adult flukes. The most pronounced anthelmintic action was exerted by DHA, whose half-maximal inhibitory concentrations (IC50) of 1.9 (NEMs) and 2.02 µg/mL (adult flukes) were lower than those of PZQ (0.56 and 0.25 µg/mL, respectively). In contrast to PZQ, the effects of DHA and AS were similar when we compared the two developmental stages of O. felineus (NEMs and adult flukes). In addition, AM, AS, and especially DHA at doses of 100 µg/mL disrupted tegument integrity in adult flukes, which was not observed with PZQ. ConclusionsArtemisinin derivatives (AS, AM, and DHA) have good anthelmintic efficacy against the trematode O. felineus, and the action of these substances is comparable to (and sometimes better than) the effects of PZQ.

Evaluation of Anthelmintic Activity of Caesalpinia Pulcherrima Leaf Extract by In-Silico and In-Vitro Studies

The Greek term for "helminthes" means "worm." Human-infecting parasites fall into one of two categories: keepsakes or heirlooms. Heirlooms are parasites that have been passed down from African ancestors, while souvenirs are parasites that humans have picked up from animals through migration, evolution, and agricultural activities. These helminthic infections are the most prevalent human infectious agents in poor nations. The helminthic parasite affects around 2 billion people worldwide, or more than 25% of the total population. It is a big problem in developing nations, particularly for young people. Therefore, the goal of the current study was to extract the plant's active ingredients and test the crude extract's anti-helminthic properties. In this study, we worked on the ceasalpeniapulcherima plant with a focus on natural remedies for the treatment of helmintiasis. The chief scientist of Sri Konda Laxman Telangana State Horticulture University, Floriculture Research Station, Rajndranagar, authenticated the true nature of the plant. Water and ethanol were utilized in the extraction procedure. To find out what chemicals are in the alcoholic extract, a screening test is performed on it. The flavonoid test on the extract came out positive. Using Albendazole as the prescribed medication, the anthelmintic action was carried out on Indian earthworms. In Indian earth worms, we found that alcoholic extract exhibits good antihelmintic action. By Insilco studies we found the interaction of flavanoids with antihelmentic proteins. Key Word: Helminthes, ceasalpeniapulcherima, flavonoids, extractioin, in silico study.

Pumpkin seeds (Cucurbita pepo subsp. ovifera) decoction promotes Trichinella spiralis expulsion during intestinal phase via “Weep and Sweep” mechanism

Trichinosis is a zoonotic disease of communal health concern as it instigated human outbreaks in several countries. Besides, the development of resistance, traditional therapy has numerous antagonistic effects. Thereby, finding efficient natural alternatives is required. In comparison to albendazole, this study evaluated the impact of pumpkin decoction on Trichinella spiralis in experimentally infected mice. The anthelmintic action of pumpkin decoction (500 mg/kg) was determined using T. spiralis infected mice in enteric phase for 5 days. Pumpkin decoction anthelmintic activity fortified by mixing with honey (1:1). Pumpkin decoction and Pumpkin decoction-honey mixture were evaluated by comprising with reference drug, albendazole (50 mg/kg). The T. spiralis adult count was significantly lower in all treated groups, with the pumpkin decoction-honey mixture showing the largest reduction (83.2%) when compared to the infected group (P ≤ 0.001). The intestinal histological changes and the level of COX-2 expression in the intestinal tissue were both significantly reduced in the same group. The pumpkin decoction improved the immune response, as evidenced by a significant decrease in nitric oxide (NO) and tumor necrosis factor (TNF-α) and a significant increase in the expression of the transforming growth factor (TGF-1β) and interleukin-17 (IL-17). The pumpkin decoction's anthelmintic action was facilitated by the TGF-1β and IL-17-driven Weep and Sweep mechanism. Both administration of pumpkin decoction beside honey showed the best treatment group that resulted in high infection reduction besides amelioration of biochemical markers and restoration of histological to normal state. In conclusion, pumpkin decoction is highly effective against T. spiralis which could be a promising alternative herbal drug and the pumpkin decoction effect was higher in the case of combination with honey.

Potential of plant extracts to inhibit egg hatching and migration of Haemonchus contortus larvae

Gastrointestinal parasitosis constitutes important health problems, aggravating throughout the years, due to inadequate use of drugs for its control, causing anthelmintic resistance. The objective of the present study was to evaluate the potential of Azadirachta indica (AZA) and Moringa oleifera (MOR) extracts in inhibiting egg hatching and in vitro migration of Haemonchus contortus larvae. A completely randomized design of experiment was carried out, where 15 treatments and a dimethyl sulfoxide negative control were evaluated: MOR-75, MOR-50, MOR-25, MOR-12.5, and MOR-6.25 mg/mL; AZA-75, AZA-50, AZA-25, AZA-12.5, and AZA-6.25 mg/mL; and thiabendazole (TBZ) TBZ-200, TBZ-100, TBZ-40, TBZ-20, TBZ-10 µg/mL. The extracts did not inhibit egg hatching capacity; however, we observed a motility decrease of up to 100% in L1 larvae. The extracts affected larval migration (P < 0.0020) compared with controls (dimethyl sulfoxide and TBZ), with migration rate inhibition above 65 %. Probit analysis showed that the median effective concentrations were 60.41 and 65.69 mg/mL for M. oleifera and A. indica, respectively. The in vitro results suggest that the water extract of both plants has an anthelmintic action against gastrointestinal nematode larvae.

Network pharmacology and molecular docking study-based approach to explore mechanism of benzimidazole-based anthelmintics for the treatment of lung cancer

Emerging studies have reported the potential anticancer activity of benzimidazole-based anthelmintics (BBA) against lung cancer (LC). However, mechanism underlying the anticancer activity of BBA is unclear. Therefore, in the current study, network pharmacology and molecular docking-based approach were used to explore the potential molecular mechanism for the treatment of LC. The potential targets for BBA were obtained from multiple databases including SwissTargetPrediction, Drug Bank, Therapeutic Target Database, and Comparative Toxicogenomics Database while LC targets were collected from DisGeNet gene discovery platform, Integrated Genomic Database of NSCLC, Catalogue of Somatic Mutations in Cancer and Online Mendelian Inheritance in Man database. Protein-protein interaction (PPI) diagram of common targets was constructed using STRING online platform. Topological analysis was performed using Cytoscape and gene enrichment analysis was conducted using FunRich software. Highest degree targets were then confirmed using molecular docking and molecular dynamics simulations. The BBA were prioritized according to their S scores, with ricobendazole ranking highest followed by flubendazole, fenbendazole, mebendazole, triclabendazole, albendazole, oxibendazole, parbendazole, thiabendazole and oxfendazole. The potential targets of BBA identified using topological analysis and molecular docking were found to be CCND1 (cyclin D1), EGFR (Epidermal Growth Factor Receptor), ERBB2 (Erb-B2 Receptor Tyrosine Kinase 2/CD340), PTGS2 (Prostaglandin-endoperoxide synthase 2), and SRC (Proto-oncogene tyrosine-protein kinase). Furthermore, molecular dynamics confirmed that CCND1 and EGFR are the potential targets of ricobendazole for the treatment of LC. BBA can be further explored as a therapeutic strategy for the treatment of lung cancer under in vitro and in vivo studies. Communicated by Ramaswamy H. Sarma

Potent anthelmintic activity of a colloidal nano-silver formulation (Silversol®) against the model worm Caenorhabditis elegans

ObjectiveIn the background of a very small number of effective anthelmintics available today with a narrow activity spectrum, and a rise in resistance against them among parasitic helminths, there is an urgent need for discovery of novel broad-spectrum anthelmintics displaying no or minimal toxicity towards the host. Silver being used since centuries for therapeutic purposes and considered safe for human consumption, we investigated anthelmintic activity of a colloidal nanosilver formulation Silversol®. Anthelmintic efficacy of the test formulation was assayed employing the nematode Caenorhabditis elegans as a model worm through a live-dead count.ResultsSilversol® exerted anthelmintic action superior to one of the positive controls (Benzimidazole), and almost at par to another positive control (Ivermectin). At concentrations ≥ 2 ppm, it could kill all the worms present in the experimental well. Lower concentrations of silver were found to have a cuticle-damaging action on worms. Further investigation is warranted to assess whether Silversol® can exert similar potent activity against different species of parasitic helminths, and elucidate the underlying molecular mechanisms of action.

Optimal schemes for the use of anthelmintics at gastrointestinal strongylatosis of young cattle

The purpose of the research is to study the rational terms for the use of anthelmintics in case of gastrointestinal strongylatoses of young cattle in Nonchernozem zone of the Russian Federation.Materials and methods. The timing of the onset of re-isolation of nematode eggs with feces of cattle after the use of some anthelmintics was determined on 50 black-motley calves weighing 117–130 kg, spontaneously infected with gastrointestinal strongylates. The optimal schemes for the use of anthelmintics in case of gastrointestinal strongylatosis of young cattle were studied on 109 calves of the first year of grazing in the pasture period of 2010. After numbering, the animals were weighed and divided into experimental and control groups. Each group was divided into two subgroups of 15–17 heads. Animals of all groups from May 3 to October 10, 2018 were grazed together in one herd on a pasture where cattle infected with strongylates had previously grazed. Levamisole from the group of imidothiazoles, Alben from the group of benzimidazoles and Aversect-2 from the group of macrocyclic lactones were tested. The drugs were used in therapeutic doses: levamisole at a dose of 7.5 mg/kg by AS intramuscularly, Alben at a dose of 7.5 mg/kg with concentrated feed, and Aversect-2 at a dose of 0.2 mg/kg subcutaneously once. Levamisole was administered to the animals of the first group on the 6th, 10th and 14th weeks of grazing, and to the animals of the second group - on the 6th and 10th weeks of grazing. Alben was used on young cattle of the third group on the 6th, 11th and 16th weeks, and on the animals of the fourth group - on the 6th and 11th weeks of grazing. Aversect-2 was administered to cattle of the fifth group at the 6th, 13th and 20th weeks of grazing, and to the animals of the sixth group – at the 6th and 13th weeks. Animals of the 7th group did not receive the drug and served as control. Fecal samples were taken from animals of all groups on a monthly basis and investigated by a quantitative method in order to detect eggs of gastrointestinal strongylate. The results obtained were statistically processed using the computer program Microsoft Excel.Results and discussion. Tests of drugs for gastrointestinal strongylatoses of young cattle showed different persistence of anthelmintic action. After deworming the animals, single eggs of strongylate in their faeces were again detected 4 weeks after the administration of Levamisole, 5 weeks after the administration of Alben, and 7 weeks after the administration of Aversect-2. The most rational scheme for deworming young cattle with gastrointestinal strongylatoses is the use of Aversect-2 on the 6th and 19th weeks of grazing.

In vitro Anthelmintic Activity of Vitis vinifera Leaf Extract on Dicrocoelium dendriticum

Background: Helminth parasites of ruminants are a major factor limiting animal production in many parts of the world. Dicrocoelium dendriticum, causes liver fluke disease in ruminants and is of zoonotic and economic importance. D. dendriticum lives in the adult stage, in the bile ducts and gallbladders of cattle, sheep, goats and pigs. Dicrocoeliasis causes severe pathological changes in the liver and bile system such as abscesses, granulomas and fibrosis. The objective of this study was to identify the anthelmintic activity of Vitis vinifera leaf extract against D. dendriticum and comparison with some drugs used against internal parasites. Methods: A study was performed to evaluate the anthelmintic activity of the methanolic extract of V. vinifera leaf against D. dendriticum using an adult worm motility test. Six graduated concentrations of extract (25, 50, 75, 100, a125 and 150 mg/ml), were tested at different periods and changes over time in the viability of worms were registered for 2, 4 and 6 hr. Normal RPMI-1640 medium and Albendazole were used as negative and positive control, respectively. Result: A significant elevation in mean inhibition of adult worm motility beginning 2 h post-exposure with 150 mg/ml of V. vinifera leaf extract and Albendazole. After 4 h of the exposure time, Albendazole and concentrations of 125 and 150 mg/ml of the extract resulted in significantly higher inhibited motility compared to the remaining concentrations below 125 mg/ml. The lower concentration (25 mg/ml) was significantly more lethal than the negative control (RPMI-1640 medium) at 6 h of exposure. Within 6 h after exposure, concentrations of 100, 125 and 150 mg/ml of V. vinifera leaf extract were more effective against adult worms than the remaining concentrations below 100 mg/ml. As the concentration and duration of exposure increased, the mortality of adult worms and also increased death for all parasites (P less than 0.05). Each concentration damages the tegument and muscles of D. dendriticum. The increase in concentration is proportional to the increase in damage to the integument. The results of this investigation demonstrated the anthelmintic action of V. vinifera leaf extract.

Endofungal bacteria boost anthelminthic host protection with the biosurfactant symbiosin.

Effective protection of soil fungi from predators is crucial for their survival in the niche. Thus, fungi have developed efficient defence strategies. We discovered that soil beneficial Mortierella fungi employ a potent cytotoxin (necroxime) against fungivorous nematodes. Interestingly, this anthelminthic agent is produced by bacterial endosymbionts (Candidatus Mycoavidus necroximicus) residing within the fungus. Analysis of the symbiont's genome indicated a rich biosynthetic potential, yet nothing has been known about additional metabolites and their potential synergistic functions. Here we report that two distinct Mortierella endosymbionts produce a novel cyclic lipodepsipeptide (symbiosin), that is clearly of bacterial origin, but has striking similarities to various fungal specialized metabolites. The structure and absolute configuration of symbiosin were fully elucidated. By comparative genomics of symbiosin-positive strains and in silico analyses of the deduced non-ribosomal synthetases, we assigned the (sym) biosynthetic gene cluster and proposed an assembly line model. Bioassays revealed that symbiosin is not only an antibiotic, in particular against mycobacteria, but also exhibits marked synergistic effects with necroxime in anti-nematode tests. By functional analyses and substitution experiments we found that symbiosin is a potent biosurfactant and that this particular property confers a boost in the anthelmintic action, similar to formulations of therapeutics in human medicine. Our findings illustrate that "combination therapies" against parasites already exist in ecological contexts, which may inspire the development of biocontrol agents and therapeutics.

Atividade anti-helmíntica in vitro de nitazoxanida em comparação com praziquantel contra Eurytrema coelomaticum

ABSTRACT: Eurytrema coelomaticum are trematodes that parasitize cattle, buffaloes, goats, sheep, and camelids, and, accidentally, humans and cats. The affected animal usually has a subclinical pancreatic disease, but the damage caused to health and animal production is underestimated. Currently, praziquantel is the unique drug proven effective against this parasite, but no formulations containing this drug are available for ruminants in Brazilian market. The objective of the present study was to separately evaluate in vitro anthelmintic activity of praziquantel (PZQ) and nitazoxanide (NTZ) against E. coelomaticum by assessing motility after 3, 12, and, 15 hours of incubation and subsequent histopathological examination of the parasites. E. coelomaticum specimens were obtained from the pancreas of naturally infected cattle, collected from animals slaughtered in the city of Concordia-SC, Brazil. The specimens were incubated in plates with a culture medium (n = 60 per group), with 80µg/ml of PZQ (positive control group), or 61,5µg/ml of NTZ (treated group), or UNT (untreated control). After 12 hours of incubation all parasites of the NTZ and PZQ groups were motionless or dead, while in the negative control group, 91.67% (55/60) presented normal motility after 15 hours of incubation (P < 0.001). Histopathological examination showed severe damage in vitellogenic gland, intestine, parenchyma, integument, and testicle in both treated and positive control groups. It was concluded that PZQ and NTZ showed in vitro anthelmintic action against the parasite Eurytrema coelomaticum, as they caused significant lesions in the evaluated organs and reduced the parasite’s motility.

Pharmacognostical, Phytochemical and In-Vitro Anthelmintic Activity of Cassia roxburghii Seed and Boerhaavia diffusa Root against Pheritima posthuma Model

Plants have served human beings as a natural source for treatments and therapies from ancient times, amongst them medicinal herbs have gain attention because of its wide use and less side effects. In the recent years plant research has increased throughout the world and a huge amount of evidences have been collected to show immense potential of medicinal plants used in various traditional systems. The objective of this study was to investigate pharmacognostical, phytochemical features and anthelmintic action of Cassia roxburghii (seeds) and Boerhaavia diffusa (roots). The different pharmacognostical parameters were evaluated as per standard protocols with some modifications. Qualitative analysis of various phytochemical constituents was determined by the well-known test protocol available in the literature. Phytochemical analysis revealed the presence of alkaloids, glycosides, flavonoids, steroids, saponins, triterpenoits and carbohydrate. Three different doses (10, 25, & 50 mg/ml) of each extracts were studied, which involves the determination of time of paralysis and time of death of worm. All the extracts showed a dose dependent increase in the anthelmintic action. Out of all the extracts, the methanol extracts showed highest activity in both plants followed by ethyl and aqueous extracts. At the concentration of 50 mg/ml the methanol showed a remarkable anthelmintic activity which was even greater than the standard drug (Albendazole) at the same concentration.
 Keywords: Cassia roxburghii, Boerhaavia diffusa, Pheretima posthuma, Anthelmintic action, Paralysis, Albendazole.

Elucidation of the nematicidal mode of action of grammicin on Caenorhabditis elegans

Grammicin (Gra) is derived from the endophytic fungus Xylaria grammica EL000614 and shows nematicidal activity against the devastating root-knot nematode Meloidogyne incognita in-vitro, in planta, and in-field experiments. However, the mechanism of the nematicidal action of Gra remains unclear. In this study, Gra exposure to the model genetic organism Caenorhabditis elegans affected its L1, L2/3, L4, and young adult stages. In addition, Gra treatment increased the intracellular reactive oxygen species (ROS) levels of C. elegans and M. incognita. Molecular docking interaction analysis indicated that Gra could bind and interact with GCS-1, GST-4, and DAF-16a in order of low binding energy, followed by SOD-3, SKN-1, and DAF-16b. This implies that the anthelmintic action of Gra is related to the oxidative stress response. To validate this mechanism, we examined the expression of the genes involved in the oxidative stress responses following treatment with Gra using transgenic C. elegans strains such as the TJ356 strain zIs356 [daf-16p::daf-16a/b::GFP + rol-6 (su1006)], LD1 ldIs7 [skn-1p::skn-1b/c::GFP + rol-6 (su1006)], LD1171 ldIs3 [gcs-1p::GFP + rol-6 (su1006)], CL2166 dvIs19 [(pAF15) gst-4p::GFP::NLS], and CF1553 strain muIs84 [(pAD76) sod-3p::GFP + rol-6 (su1006)]. Gra treatment caused nuclear translocation of DAF-16/FoxO and enhanced gst-4::GFP expression, but it had no change in sod-3::GFP expression. These results indicate that Gra induces oxidative stress response via phase II detoxification without reduced cellular redox machinery. Gra treatment also inhibited the nuclear localization of SKN-1::GFP in the intestine, which may lead to a condition in which oxidative stress tolerance is insufficient to protect C. elegans by the inactivation of SKN-1, thus inducing nematode lethality. Furthermore, Gra caused the mortality of two mutant strains of C. elegans, CB113 and DA1316, which are resistant to aldicarb and ivermectin, respectively. This indicates that the mode of action of Gra is different from the traditional nematicides currently in use, suggesting that it could help develop novel approaches to control plant-parasitic nematodes.