Adult Worms Research Articles

Gene-guided identifications of a structure-chimeric cyclotide viphi I from Viola philippica: Potential functions against cadmium and nematodes

The cyclic peptides, cyclotides, are identified mostly with 29–31-aa (amino acid residues) but rarely with ≥ 34-aa in plants. Viola philippica is a well-known medicinal plant but a rare metallophyte with cyclotides. A hypothesis was hence raised that the potential novel 34-aa cyclotide of Viola philippica would clearly broaden the structural and functional diversities of plant cyclotides. After homology-cloning the cyclotide precursor gene of VpCP5, a 34-aa cyclotide (viphi I) was identified to be larger than 22 other known cyclotides in V. philippica. It had a chimeric primary structure, due to its unusual loop structures (8 residues in loop 2 and 6 residues in loop 5) and aa composition (3 E and 5 R), by using phylogenetic analyses and an in-house cyclotide analysis tool, CyExcel_V1. A plasmid pCYC-viphi_I and a lab-used recombinant process were specially constructed for preparing viphi I. Typically, 0.12 or 0.25 mg ml−1 co-exposed viphi I could significantly remain cell activities with elevating Cd2+-exposed doses from 10−8 to 10−6 mol l−1 in MCF7 cells. In the model nematode Caenorhabditis elegans, IC50 values of viphi I to inhibit adult ratios and to induce death ratios, were 184.7 and 585.9 µg ml−1, respectively; the median lifespan of adult worms decreased from 14 to 2 d at viphi I doses ranging from 0.05 to 2 mg ml−1. Taken together, the newly identified viphi I exhibits functional potentials against cadmium and nematodes, providing new insights into structural and functional diversity of chimeric cyclotides in plants.

Diagnosis of Experimental Trichinosis by Nano-Based ELISA and Nano-Based Latex Agglutination Test Compared with Traditional Sandwich ELISA

Abstract Human trichinellosis is a serious foodborne parasitic zoonosis. The main route for contracting the infection is eating raw or undercooked meat, especially pork containing a sufficient load of infective viable larvae. Early diagnosis is crucial before the larval encystation in skeletal muscles, so detection of circulating antigens can be helpful. The objective of the present study was to evaluate the novel Nanomagnetic beads based-ELISA (NMB-ELISA), and Nanomagnetic beads based latex agglutination test (NMB-LAT) for the diagnosis of experimental trichinellosis compared to sandwich ELISA through the detection of Trichinella spiralis adult worm crude extract antigen AW-CEA in serum. The study included thirty-eight (38) mice classified into 3 groups; T. spiralis infection group (GI) which was equally subdivided into 5 subgroups according to the time of sacrifice (6, 8, 10, 12, and 14 dpi), other parasitic infections group (GII) and healthy control group (GIII). T. spiralis AW-CEA was prepared from the adult worms through homogenization and ultra-sonication and then used to produce anti- T. spiralis IgG-pAbs in rabbits which were subsequently utilized to detect AW-CEA in serum samples by sandwich ELISA, NMB-ELISA, and NMB-LAT. Using NMB-ELISA, T. spiralis AW-CEA was detected in sera collected at 6 and 8 dpi, with a sensitivity of 50%, and 75%, respectively, and a specificity of 100%. Meanwhile, sandwich ELISA and NMB-LAT couldn’t detect the antigen at the same time intervals. Both ELISA formats were able to detect the antigen in samples collected at 10, 12, and 14 dpi with a sensitivity of 100% for NMB-ELISA, and 25%, 75%, and 100% respectively, for sandwich-ELISA. On the other hand, NMB-LAT couldn't detect T. spiralis AW-CEA until 12 dpi with a sensitivity of 50% and specificity of 75%. In conclusion, the NMB-ELISA is a promising sensitive technique for early and specific diagnosis of acute trichinellosis in an animal model. Thus, it can be used as a potential alternative to the standard ELISA to obtain confident results. In addition, the use of NMB-LAT for AW-CEA detection could be a valuable screening diagnostic technique in field surveys.

The Secretome of Adult Murine Hookworms Is Shaped by Host Expression of STAT6.

Co-evolutionary adaptation of hookworms with their mammalian hosts has been selected for immunoregulatory excretory/secretory (E/S) products. However, it is not known whether, or if so, how host immunological status impacts the secreted profile of hematophagous adult worms. This study interrogated the impact of host Signal transducer and activator of transcription 6 (STAT6) expression during the experimental evolution of hookworms through the sequential passage of the life cycle in either STAT6 deficient or WT C57BL/6 mice. Proteomic analysis of E/S products by LC-MS showed increased abundance of 15 proteins, including myosin-3, related to muscle function, and aconitate hydratase, related to iron homeostasis. However, most E/S proteins (174 of 337 unique identities) were decreased, including those in the Ancylostoma-secreted protein (ASP) category, and metallopeptidases. Several identified proteins are established immune-modulators such as fatty acid-binding protein homologue, cystatin, and acetylcholinesterase. Enrichment analysis of InterPro functional categories showed down-regulation of Cysteine-rich secretory proteins, Antigen 5, and Pathogenesis-related 1 proteins (CAP), Astacin-like metallopeptidase, Glycoside hydrolase, and Transthyretin-like protein groups in STAT6 KO-adapted worms. Taken together, these data indicate that in an environment lacking Type 2 immunity, hookworms alter their secretome by reducing immune evasion proteins- and increasing locomotor- and feeding-associated proteins.

Mass Spectrometry Identifies Taenia solium Proteins in Sera of Patients With and Without Parenchymal Neurocysticercosis.

Neurocysticercosis (NCC), a major cause of global acquired epilepsy, results from Taenia solium larval brain infection. T. solium adult worms release large numbers of infective eggs into the environment contributing to high levels of exposure in endemic areas. This study identifies T. solium proteins in the sera of individuals with and without NCC using mass spectrometry to examine exposure in endemic regions. Forty-seven patients (18-51 years), 24 parenchymal NCC (pNCC), 8 epilepsy of unknown aetiology, 7 glioma, 8 brain tuberculoma, and 7 healthy volunteers were studied. Trypsin digested sera were subject to liquid chromatography-tandem mass spectrometry and spectra of 375-1700 m/z matched against T. solium WormBase ParaSite database with MaxQuant software to identify T. solium proteins. Three hundred and nineteen T. solium proteins were identified in 87.5% of pNCC and 56.6% of non-NCC subjects. Three hundred and four proteins were exclusive to pNCC sera, seven to non-NCC sera and eight in both. Ten percent, exhibiting immune-modulatory properties, originated from the oncosphere and cyst vesicular fluid. In conclusion, in endemic regions, T. solium proteins are detected in sera of individuals with and without pNCC. The immunomodulatory nature of these proteins may influence susceptibility and course of infection.

First Molecular Identification and Clinical Presentation of Crenosomosis in a Dog from Slovakia

PurposeCrenosoma vulpis (Dujardin,1845) is a lungworm which has spread worldwide in canines and is associated with upper respiratory infections. In a majority of cases, the infections are accompanied with chronic cough. Diagnosis of lungworms is often underdiagnosed and can be misinterpreted as other respiratory diseases.MethodsThe Small Animal Clinic of the University Veterinary Hospital admitted an 11-month-old dog presented with persistent cough associated with difficulty in breathing and even asphyxia. Based on clinical symptoms, the patient underwent radiological and bronchoscopic examination. Bronchoscopy revealed the presence of lungworms obturating the branches of the tracheobronchial tree. Larvae were collected by bronchoscopic lavage and subjected to parasitological and molecular examination.ResultsMicroscopic detection and morphological identification of the worms removed during the bronchoscopy confirmed the presence of female adult worms. The subsequent molecular characterisation of the mitochondrial (cytochrome c oxidase subunit I gene (cox1) and 12S ribosomal DNA (rDNA)), nuclear (18S rDNA) genes, as well as the analysis of the second internal transcribed spacer (ITS-2) region of the ribosomal DNA, confirmed the Crenosoma vulpis species. Faecal samples were processed using the Baermann method, which confirmed the presence of the larval stage 1 of C. vulpis. The therapy with fenbendazole at a dose of 50 mg/kg of live weight once daily for the period of 7 days was initiated for the patient.ConclusionThis paper presents the first molecularly confirmed clinical case of a Crenosoma vulpis infection in an 11-month-old female dog of the Miniature Schnauzer breed in Slovakia.

Efficacy and safety of single-dose artesunate plus sulfalene/pyrimethamine combined with praziquantel for the treatment of children with Schistosoma mansoni or Schistosoma haematobium in western Kenya: a randomised, open-label controlled trial

BackgroundReliance on praziquantel for the treatment and control of schistosomiasis is likely to facilitate the emergence of drug resistance. Combination therapy targeting adult and juvenile schistosome worms is urgently needed to improve praziquantel efficacy and delay the potential development of drug resistance. We assessed the efficacy and safety of single-dose praziquantel combined with single-dose artesunate plus sulfalene-pyrimethamine in the treatment of Kenyan children with schistosomiasis.MethodsThis was an open-label, randomised clinical trial involving 426 school-aged children (7–15 years old) diagnosed with Schistosoma mansoni (by Kato-Katz) or S. haematobium (by urine filtration). They were randomly assigned (1:1:1) to receive a single dose of praziquantel (40 mg/kg), a single dose of artesunate plus sulfalene-pyrimethamine (12 mg/kg artesunate) or combination therapy using a single dose of praziquantel (40 mg/kg) combined with a single dose of artesunate plus sulfalene-pyrimethamine (12 mg/kg artesunate). The primary outcome was cure and egg reduction rates at 6 weeks post-treatment in the available case population. Adverse events were assessed within 3 h after treatment.ResultsOf the 426 children enrolled, 135 received praziquantel, 150 received artesunate plus sulfalene-pyrimethamine, and 141 received combination therapy. Outcome data were available for 348 (81.7%) children. For S. mansoni-infected children (n = 335), the cure rates were 75.6%, 60.7%, and 77.8%, and the egg reduction rates were 80.1%, 85.0%, and 88.4% for praziquantel, artesunate plus sulfalene-pyrimethamine, and combination therapy, respectively. For S. haematobium-infected children (n = 145), the corresponding cure rates were 81.4%, 71.1%, and 82.2%, and the egg reduction rates were 95.6%, 97.1%, and 97.7%, respectively. Seventy-one (16.7%) children reported mild-intensity adverse events. The drugs were well tolerated and no serious adverse events were reported.ConclusionsA single oral dose of praziquantel combined with artesunate plus sulfalene-pyrimethamine cured a high proportion of children with S. haematobium but did not significantly improve the treatment efficacy for either urinary or intestinal schistosomiasis. Sequential administration of praziquantel and artesunate plus sulfalene-pyrimethamine may enhance the efficacy and safety outcomes.Graphical abstract

Protein Concentration, Anthelmintic Activity, and Microbial Contamination of the Laboratory-Produced Chitosan-Encapsulated Bromelain Batches

Bromelain has been shown to have potential as an anthelmintic for controlling livestock nematodes, such as Haemonchus (H.) contortus. The present study aimed to evaluate the in vitro quality of the laboratory-produced nanoencapsulated bromelain (NEB) and its activity against H. contortus. The acid-base extraction method was employed to extract four different batches of bromelain from the peels of fully ripened pineapples. It was encapsulated in chitosan to form the nano-encapsulated bromelain complex. Standard biochemical methods were employed to determine the bromelain concentration, protein concentration, in vitro anthelmintic activity against various stages of H. contortus (egg, larva, adult), and bacteria contamination for the four NEB batches. The mean concentration of extracted bromelain was 4.3 mg/ml in all four batches. There were no variations in the protein concentrations between the batches of NEB, which ranged from 1,090 mg/ml to 1.205 mg/ml. Although there were no significant differences in different batches, a variation in NEB inhibitory concentration (IC50) was observed according to the different parasitic stages. The highest activity was for adult worms (LC50 =0.2454 ± 0.05 mg/ml), followed by the eggs (IC50 = 0.3 ± 0.07 mg/ml), and the larval stage (IC50 =0.9 ± 0.45 mg/ml). Despite the identification of certain bacterial species in the raw pineapple extract, the final product of all four batches of NEB remained free from any bacterial contamination. The current study indicated that NEB's concentration, protein concentrations, and anthelmintic activity did not vary significantly across the different batches of NEB. Additionally, the encapsulation process ensured that the final product was free of bacterial contamination and thus safe for use in animals.

A review of moxidectin vs. other macrocyclic lactones for prevention of heartworm disease in dogs with an appraisal of two commercial formulations.

Macrocyclic lactones (MLs) are the only drug class currently licensed for heartworm disease prophylaxis. Macrocyclic lactones kill third- and fourth-stage larvae of Dirofilaria immitis, thus preventing the development of adult worms in dogs, which are responsible for heartworm disease, a potentially life-threatening condition. Despite considerable overlap in terms of endectocide spectrum, several important differences distinguish moxidectin from other MLs. Moxidectin has beneficial pharmacokinetic characteristics, such as a longer half-life and greater tissue distribution compared to ivermectin. Additionally, moxidectin has a greater margin of safety compared to ivermectin in dogs with ABCB1 (previously MDR1) gene-defect, which is commonly recognized in collies and other breeds. Multiple laboratory studies have shown that moxidectin is more effective than other commonly used heartworm preventives against resistant strains of D. immitis. This improved efficacy benefits individual dogs and helps reduce the risk of spreading resistant strains within the community. Despite the presence of proven resistant strains in the United States, non-compliance with preventive measures remains a major factor contributing to the diagnosis of heartworm disease in dogs. In retrospective analyses, the oral moxidectin combination product Simparica Trio® (sarolaner, moxidectin, and pyrantel) was associated with increased compliance, resulting in more time of protection compared to dogs receiving flea/tick and heartworm preventive products separately. Compliance with the extended-release moxidectin injectables ProHeart® 6 and ProHeart® 12 was higher than with monthly heartworm preventives, as they provide 6 months or a full year of protection with one single injection, respectively, and revenues remain in the veterinary clinics as injectable moxidectin cannot be sourced through online retailers.

Evaluation of anthelmintic activity of the essential oils from leaves and rind of Aegle marmelos (Linn.) Correa ver. Bel of family Rutaceae of Pakistan

In this study ‘anthelmintic activity of essential oils (EOs), of leaves and rind of Aegle marmelos (Linn.) Correa ver. Bel was evaluated. ‘Adult motility assay’ was employed using Haemoncus contortus adult worms. EO was applied at three levels viz. 10, 35 and 50µL/10 ml in phosphate buffer solution (PBS) plus 10µL of tween 20 (as carrier/emulsifier). Levamisole was used as a positive control at 0.55 mg/ml concentration. Each concentration of essential oils obtained from different plant parts exhibited varied anthelmintic activity. The best dose dependant effect on adult mortality was by EOs from Aegel rind (R2 values 0.981) followed by EOs from leaves (R2 values 0.86.). LC50 were 34.92 and 26.65 for rind and leaves EOs respectively. Change in the color of the dead worms was an indicator parameter of the fact that the EOs might have damaged the skin of the worms or after transcutaneous penetration into the body has disrupted the circulatory system by causing constriction of blood vessels. It is concluded that EOs extracted from A. marmelos have anthelmintic properties.

Evaluation of diagnostic techniques for early detection of heartworm in experimentally infected dogs: identification of Dirofilaria immitis-derived microRNA in the initial 28 weeks post-inoculation

BackgroundDirofilaria immitis, commonly known as heartworm (HW), is a parasitic nematode transmitted by various mosquito species, leading to heartworm disease (HWD) in dogs. Diagnosis of HW typically involves antigen or microfilariae detection, or visualization of adult worms through imaging or post mortem examination. Polymerase chain reaction (PCR) and micro RNA (miRNA) detection have been explored for HW diagnosis.MethodsThree dogs, previously experimentally infected with HW, underwent blood sampling every 4 weeks for 7 months. Samples were assessed for antigen presence after heat treatment, PCR amplification, and microfilaria examination using Giemsa-stained thick smears. Additionally, whole blood aliquots underwent miRNA deep sequencing and bioinformatic analysis.ResultsHeartworm antigen was detectable after heat treatment at 20 weeks post-inoculation and via PCR at 24 weeks, with microfilariae observed in peripheral blood smears at 28 weeks. However, deep miRNA sequencing revealed that the miRNA candidate sequences are not consistently expressed before 28 weeks of infection.ConclusionsWhile ancillary molecular methods such as PCR and miRNA sequencing may be less effective than antigen detection for detecting immature larval stages in an early stage of infection, our experimental findings demonstrate that circulating miRNAs can still be detected in 28 weeks post-infection.Graphical

Distinguishing recrudescence from reinfection in lymphatic filariasis

The Global Program to Eliminate Lymphatic Filariasis (GPELF) is the largest public health program based on mass drug administration (MDA). Despite decades of MDA, ongoing transmission in some countries remains a challenge. To optimise interventions, it is critical to differentiate between recrudescence and new infections. Since adult filariae are inaccessible in humans, deriving a method that relies on the offspring microfilariae (mf) is necessary. We developed a genome amplification and kinship analysis-based approach using Brugia malayi samples from gerbils, and applied it to analyse Wuchereria bancrofti mf from humans in Côte d'Ivoire. We examined the pre-treatment genetic diversity in 269 mf collected from 18 participants, and further analysed 1-year post-treatment samples of 74 mf from 4 participants. Hemizygosity of the male X-chromosome allowed for direct inference of haplotypes, facilitating robust maternal parentage inference. To enrich parasite DNA from samples contaminated with host DNA, a whole-exome capture panel was created for W.bancrofti. By reconstructing and temporally tracking sibling relationships across pre- and post-treatment samples, we differentiated between new and established maternal families, suggesting reinfection in one participant and recrudescence in three participants. The estimated number of reproductively active adult females ranged between 3 and 11 in the studied participants. Population structure analysis revealed genetically distinct parasites in Côte d'Ivoire compared to samples from other countries. Exome capture identified protein-coding variants with ∼95% genotype concordance rate. We have generated resources to facilitate the development of molecular genetic tools that can estimate adult worm burdens and monitor parasite populations, thus providing essential information for the successful implementation of GPELF. This work was financially supported by the Bill and Melinda Gates Foundation (https://www.gatesfoundation.org) under grant OPP1201530 (Co-PIs PUF & Gary J. Weil). B.malayi parasite material was generated with support of the Foundation for Barnes Jewish Hospital (PUF). In addition, the development of computational methods was supported by the National Institutes of Health under grants AI144161 (MM) and AI146353 (MM). The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Peptide-Alkoxyamine Drugs: An Innovative Approach to Fight Schistosomiasis: "Digging Their Graves with Their Forks".

The expansion of drug resistant parasites sheds a serious concern on several neglected parasitic diseases. Our recent results on cancer led us to envision the use of peptide-alkoxyamines as a highly selective and efficient new drug against schistosome adult worms, the etiological agents of schistosomiasis. Indeed, the peptide tag of the hybrid compounds can be hydrolyzed by worm's digestive enzymes to afford a highly labile alkoxyamine which homolyzes spontaneously and instantaneously into radicals-which are then used as a drug against Schistosome adult parasites. This approach is nicely summarized as digging their graves with their forks. Several hybrid peptide-alkoxyamines were prepared and clearly showed an activity: two of the tested compounds kill 50% of the parasites in two hours at a concentration of 100 µg/mL. Importantly, the peptide and alkoxyamine fragments that are unable to generate alkyl radicals display no activity. This strong evidence validates the proposed mechanism: a specific activation of the prodrugs by the parasite proteases leading to parasite death through in situ alkyl radical generation.

Molecular insights and antibody response to Dr20/22 in dogs naturally infected with Dirofilaria repens

Subcutaneous dirofilariasis, caused by the parasitic nematode Dirofilaria repens, is a growing concern in Europe, affecting both dogs and humans. This study focused on D. repens Dr20/22, a protein encoded by an alt (abundant larval transcript) gene family. While well-documented in L3 larvae of other filariae species, this gene family had not been explored in dirofilariasis. The research involved cloning Dr20/22 cDNA, molecular characterization, and evaluating its potential application in the diagnosis of dirofilariasis. Although Real-Time analysis revealed mRNA expression in both adult worms and microfilariae, the native protein remained undetected in lysates from both developmental stages. This suggests the protein’s specificity for L3 larvae and may be related to a process called SLTS (spliced leader trans-splicing), contributing to stage-specific gene expression. The specificity of the antigen for invasive larvae positions it as a promising early marker for dirofilariasis. However, ELISA tests using sera from infected and uninfected dogs indicated limited diagnostic utility. While further research is required, our findings contribute to a deeper understanding of the molecular and immunological aspects of host-parasite interactions and could offer insights into the parasite's strategies for evading the immune system.

Ocular filariasis in the form of a conjunctival granuloma – A rare case report from non-endemic part of India

Filariasis, a predominantly lymphatic disease can rarely have ocular involvement in endemic as well as a few non-endemic zones of the Indian subcontinent. It has also been reported from countries of South East Asia. In India, Wuchereria bancrofti and Brugia malayi are the nematodes causing filariais. Ocular involvement can happen in a wide spectrum, involving lacrimal glands, conjunctiva and fornices, cornea, anterior chamber, and vitreous cavity. We report a case of adult filarial worm (B. malayi) inside a conjunctival granuloma in a lady hailing from a rural, non-endemic part of the country. The lady presented with a short history of ocular irritation and redness. Clinical examination revealed a small granuloma-like lesion with surrounding inflammation over the conjunctiva. The rest of the anterior segment as well as the posterior segment was fairly within normal limits. The history as well as clinical examination was not supportive of a parasitic infestation, leading the doctors to treat the lesion as an allergic disease. However, the lesion remained unresolved, and surgical excision was undertaken. On surgical intervention, a live worm was spotted inside the lesion. Parasitological evaluation of the worm was confirmatory of adult filaria (B. malayi). The patient responded to a combination therapy of oral diethylcarbamazine and albendazole.

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.

Description and circadian rhythms of Chandlerella sinensis Li, 1933 (Nematoda; Onchocercidae), with remarks of microfilariae effects on the host health.

During investigation of common linnet (Linaria cannabina) blood using the buffy coat method one bird with microfilariae in the blood was found. The morphometric description of adult worms corresponded to the Chandlerella sinensis. This species was found for the first time in common linnets. DNA sequences of cox1 and 28S gene fragments of adult worm recovered during necropsy was identical to that from the microfilariae in the bird blood. Phylogenetic analysis of the cox1 gene fragment clustered this parasite with Chandlerella quiscali. Histological examination revealed the presence of microfilariae in the lumen of small capillaries and other blood vessels in different organs, but no inflammations were notice. The greatest number of microfilariae was in the lungs. Even if there was no inflammation, but vessels associated with the lungs were markedly distended with blood, parabronchial walls were thickened and, in some cases, almost completely obstructing the lumen. The large number of microfilariae in lungs indicates possible disturbance of gas exchange in the lungs adversely affected the ability of the bird to exercise and made breathing difficult at rest. The investigation of circadian rhythm of the microfilariae showed that C. sinensis microfilariae in blood of common linnet were more numerous at night and morning and less numerous at midday. The survival rate of mosquitoes infected with C. sinensis microfilariae was significantly lower than that of uninfected mosquitoes.

Evaluating the Toxocara cati extract as a therapeutic agent for allergic airway inflammation.

The hygiene hypothesis suggests that early life exposure to helminth infections can reduce hypersensitivity in the immune system. The present study aims to evaluate the effects of Toxocara cati(T. cati) somatic products on allergic airway inflammation. Between 2018 and 2020, T. cati adult worms were collected from stray cats in Mashhad, Iran (31 out of 186 cats), and their somatic extract was collected. Thirty BALB/cmice were equally divided into three groups, including the OVA group (sensitized and challenged with ovalbumin), the somatic administered group (received somatic extract along with ovalbumin sensitization), and the PBS group (sensitized and challenged with phosphate buffer saline). Bronchoalveolar lavage(BAL)fluid was collected to assess the number of cells, and lung homogenates were prepared for cytokine analysis. Histopathological analysis of the lungs was performed, and inflammatory cells and mucus were detected. Cytokine levels (IL-4, IL-5, IL-10) were measured using enzyme-linked immunosorbent assay (ELISA), and ovalbumin-specific immunoglobulin E (IgE) levels were determined using a capture ELISA. The somatic group significantly decreased regarding the lung pathological changes, including peribronchiolitis, perivasculitis, and eosinophil influx, compared to the group treated with ovalbumin alone. These changes were accompanied by a decrease in proinflammatory cytokines IL-4 and IL-5 and an increase in the anti-inflammatory cytokine IL-10, indicating a shift towarda more balanced immune response. The number of inflammatory cells in the BAL fluid was also significantly reduced in the somatic group, indicating a decrease in inflammation. These preclinical findings suggest that in experimental models, T. cati somatic extract exhibits promising potential as a therapeutic agent for mitigating allergic airway inflammation. Its observed effects on immune response modulation and reduction of inflammatory cell infiltration warrant further investigation in clinical studies to assess its efficacy and safety in human patients.

Anthelmintic Activity of Red Macroalgae Acrocystis sp. and Acanthophora sp. Etanolic Extract Against Haemonchus contortus in Sheep In Vitro

Abstract This study aimed to evaluate the anthelmintic activity of ethanolic extracts from red macroalgae Acrocystis sp. and Acanthophora sp. against the nematode Haemonchus contortus found in sheep, in vitro. The adult worm motility test was conducted in a factorial completely randomized design with two factors: the type of macroalgae as the first factor and the time of observation as the second factor. A total of 100 adult female H. contortus worms were randomly divided into four treatments with five replicates each. Each replicate consisted of 5 worms placed in a petri dish containing 5 mL of 0.9% sodium chloride solution (C: negative control), 1 mg/mL ethanolic extract of Acrocystis sp. (AR), 1 mg/mL ethanolic extract of Acanthophora sp. (AC), and 0.5 mg/mL albendazole (ALB: positive control). Sodium chloride 0.9% was used as the solvent for AR, AC, and ALB. The results demonstrated that, after 24 hour, AC exhibited anthelmintic activity by inhibiting the motility of H. contortus (P<0.05), to a remaining 4.00%, compared to AR (44.00%), although it could not compete with albendazole (P<0.05). Accordingly, the ethanolic extract of red macroalgae Acanthophora sp. holds potential for further investigation as an anthelmintic agent for ruminant livestock.

Oesophagostomum stephanostomum causing parasitic granulomas in wild chimpanzees (Pan troglodytes verus) of Taï National Park, Côte d'Ivoire.

Nematodes belonging to the genus Oesophagostomum frequently infect wild chimpanzees (Pan troglodytes) across widely separated field sites. Nodular lesions (granulomas) containing Oesophagostomum are commonly seen in the abdomen of infected chimpanzees post-mortem. At Taï National Park, Côte d'Ivoire, previous studies have identified larvae of a variety of Oesophagostomum spp. in wild chimpanzee stool, based on sequencing of larval DNA, and nodular lesions associated with Oesophagostomum, identified morphologically to the genus level but not sequenced. Here we present three recent cases of parasitic granulomas found post-mortem in chimpanzees at Taï. We complement descriptions of gross pathology, histopathology and parasitology with PCR and sequencing of DNA isolated from the parasitic nodules and from adult worms found inside the nodules. In all three cases, we identify Oesophagostomum stephanostomum as the causative agent. The sequences from this study were identical to the only other published sequences from nodules in nonhuman primates-those from the wild chimpanzees of Gombe, Tanzania.

New Insights on Tools for Detecting β-Tubulin Polymorphisms in Trichuris trichiura Using rhAmpTM SNP Genotyping.

Soil-transmitted helminth (STH) infections, commonly treated with benzimidazoles, are linked to resistance through single nucleotide polymorphisms (SNPs) at position 167, 198, or 200 in the β-tubulin isotype 1 gene. The aim of this study was to establish a novel genotyping assay characterized by its rapidity and specificity. This assay was designed to detect the presence of SNPs within the partial β-tubulin gene of Trichuris trichiura. This was achieved through the biallelic discrimination at codons 167, 198, and 200 by employing the competitive binding of two allele-specific forward primers. The specificity and reliability of this assay were subsequently confirmed using Trichuris samples isolated from captive primates. Furthermore, a molecular study was conducted to substantiate the utility of the β-tubulin gene as a molecular marker. The assays showed high sensitivity and specificity when applied to field samples. Nevertheless, none of the SNPs within the β-tubulin gene were detected in any of the adult worms or eggs from the analyzed populations. All specimens consistently displayed an SS genotype. The examination of the β-tubulin gene further validated the established close relationships between the T. trichiura clade and Trichuris suis clade. This reaffirms its utility as a marker for phylogenetic analysis.