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It is not uncommon for pigeons kept for entertainment or competition to be infected with a variety of intestinal parasites, including nematodes and cestodes. Infection by these worms is a risk not only to birds but also to native wildlife and humans. The objective of this study is to assess the prevalence of helminth infections in pigeons in Zabol County, Iran, and to develop strategies for the prevention and treatment of these infections. A total of 220 pigeons were examined for the presence of parasites in their gastrointestinal tracts between 2021 and 2022. The pigeons were categorized according to gender, breed, age, medication, and feed type. The digestive tract contents were scraped and washed with warm water in order to collect intestinal helminths. A variety of techniques employed for the isolation, observation, and identification of nematodes and cestodes. Additionally,the parasites around the esophageal tissue were also examined in this study. A total of 96 positive cases of helminth infection (43.63%) were identified in the 220 samples of pigeon gastrointestinal tracts examined. In addition, 52 pigeons exhibited exclusive cestode infections, 14 had nematode infections, and 31 had co-infections. Heterakis gallinarum (H. gallinarum), Eulimdana clava (E. clava), Hadjelia truncate (H. truncate), and Ascaridia columbae (A. columbae) were among the nematodes found. Cestode species identified include Raillietina tetragona (R. tetragona), Raillietina echinobothrida (R. echinobothrida), Raillietina cesticillus (R. cesticillus), Raillietina magninumida (R. magninumida), and Cotugnia digonopora (C. digonopora). A statistical analysis of data revealed a significant correlation between helminth infection and drug consumption (p=0.001). Infection with nematodes and cestodes is also associated with pigeon age (p=0.00001). T Notably, despite the hot and dry conditions, there was a high prevalence of cestode and nematode infestations in pigeons in Zabol, Iran. It is imperative that a comprehensive control program be implemented to prevent parasite transmission to pigeons and the local avian ecosystem. [ABSTRACT FROM AUTHOR]

Fisheries play a crucial role in aquatic ecosystems, a resource with ecological and economic importance. Measuring parasite prevalence is necessary to assess these fish species' health and well-being. Within the Sistan region, this study sought to estimate the prevalence and intensity of infection with plerocercoid Ligula intestinalis (L. intestinalis) in seven fish species and identify factors such as fish weight, seasonality, and fishing location that might influence infection rates. 2800 fish from seven species were obtained in the Sistan region throughout four seasons. Fish samples from three weight categories (<200gr, 200–400gr, 400gr<) were examined parasitologically. Ligula intestinalis was detected in 96/2800 (3.4%) of fishs. There was a variation in prevalence among fish species. Alburnus charusini (A. charusini), Hypophthalmichthys molitrix (H. molitrix), Hypophthalmichthys nobilis (H. nobilis), and Schizocypris altidorsalis (S. altidorsalis) showed no infection. Schizothorax zarudnyi (S. zarudnyi) showed a prevalence of 1.5% (6/400), Cyprinus carpio (C. carpio) 15% (60/400), and Ctenopharyngodon idella (C. idella) 7.5% (30/400). It was found that infection prevalence was significantly influenced by fish weight in C. carpio and C. idella. A higher prevalence was found in fish with lower weight. Infection rates significantly fluctuated with season across species, but fishing locations had no significant difference. The present study highlights the significant prevalence of L. intestinalis infections in fish populations within Iran's Sistan region, underscoring the need for ongoing monitoring and research to inform effective fishery management strategies. Future studies should focus on long-term monitoring efforts and consider the broader global context of fish parasitology to understand parasite-host interactions better and inform conservation measures worldwide. [ABSTRACT FROM AUTHOR]

Background and Aim: Schima wallichii Korth., commonly known as the needlewood tree (family Theaceae) has therapeutic uses in traditional Mizo medicine for human helminthiasis and serves as a balm against ectoparasites in animals. Although the medicinal properties have been studied experimentally, its use as a traditional anthelmintic remains unexplored. This study aimed to analyze the chemical components and antiparasitic activity of S. wallichii. Materials and Methods: The chemical analysis of S. wallichi bark extracts was conducted focusing on the secondary metabolites using petroleum ether, chloroform, and methanol. Gas chromatography-mass spectrometry (GC-MS) was used to identify the specific compounds. An anthelmintic susceptibility test was carried out against Raillietina tetragona, intestinal cestode parasite of fowl. Results: The methanol extract yielded the highest concentrations of alkaloids, carbohydrates, glycosides, sterols, saponins, and tannins among all the extracts. Sterols were the most abundant compounds in all extracts, with flavonoids being absent. Secondary metabolites were largely absent in the petroleum ether and chloroform extracts. The GC-MS data identified cholest-22-ene-21-ol as the major steroid component. The cestode parasite was inhibited in a concentration-dependent manner by the plant extract. The plant extract’s anthelmintic activity was evident through observable damage to the parasite’s outer structure. Conclusion: Phytosterols in S. wallichii bark are responsible for its anthelmintic properties. The mechanism and pharmaceutical properties of the anthelmintic molecule require further exploration.

Background and Aim: Schima wallichii Korth., commonly known as the needlewood tree (family Theaceae) has therapeutic uses in traditional Mizo medicine for human helminthiasis and serves as a balm against ectoparasites in animals. Although the medicinal properties have been studied experimentally, its use as a traditional anthelmintic remains unexplored. This study aimed to analyze the chemical components and antiparasitic activity of S. wallichii. Materials and Methods: The chemical analysis of S. wallichi bark extracts was conducted focusing on the secondary metabolites using petroleum ether, chloroform, and methanol. Gas chromatography-mass spectrometry (GC-MS) was used to identify the specific compounds. An anthelmintic susceptibility test was carried out against Raillietina tetragona, intestinal cestode parasite of fowl. Results: The methanol extract yielded the highest concentrations of alkaloids, carbohydrates, glycosides, sterols, saponins, and tannins among all the extracts. Sterols were the most abundant compounds in all extracts, with flavonoids being absent. Secondary metabolites were largely absent in the petroleum ether and chloroform extracts. The GC-MS data identified cholest-22-ene-21-ol as the major steroid component. The cestode parasite was inhibited in a concentration-dependent manner by the plant extract. The plant extract's anthelmintic activity was evident through observable damage to the parasite's outer structure. Conclusion: Phytosterols in S. wallichii bark are responsible for its anthelmintic properties. The mechanism and pharmaceutical properties of the anthelmintic molecule require further exploration. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Anatolian Environmental & Animal Science is the property of Bulent Verep and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The tapeworms of Moniezia spp. are heteroxenous parasites and their adult forms occur in ruminants' alimentary tract. They steal a significant portion of hosts' nourishment initiating monieziasis, thereby inflicting economic losses in animal rearing. Despite their high economic importance, the molecular characterization and taxonomic status of these parasites have remained poorly understood. In the present study, cestodes were isolated from the sheep and goats' intestines and were stained with Gower's carmine. Upon careful evaluation of morphological characters, 2 species Moniezia denticulata and Moniezia expansa were identified. The genomic DNA was extracted and polymerase chain reaction (PCR) amplified targeting regions of mitochondrial cytochrome c oxidase subunit 1 (cox1), small subunit ribosomal RNA (SSU rRNA) and internal transcribed spacer 1–5.8S rRNA (ITS1–5.8S rRNA) genes followed by sequencing. The partial sequences of cox1, SSU rRNA and ITS1–5.8S rRNA genes of M. denticulata generated in the present study revealed that even though they share high similarities with M. benedeni (93.2% cox1; 92.6% SSU rRNA; 84.70% ITS1–5.8S rRNA) and M. expansa (88.85% cox1; 92.27% SSU rRNA; 81.70% ITS1–5.8S rRNA), they are not identical to them. In the maximum likelihood phylogenetic trees, M. denticulata and M. expansa consistently appeared as distinct species from each other. The high values of pairwise divergence between these 2 species collected in the present study confirmed their separate identity. The present study reports the first molecular characterization of M. denticulata with reference to M. expansa infecting sheep and goats in India.

Parasites with complex life cycles often manipulate the phenotype of their intermediate hosts to increase the probability of transmission to their definitive hosts. Infection with Anomotaenia brevis, a cestode that uses Temnothorax nylanderi ants as intermediate hosts, leads to a multiple‐fold extension of host lifespan and to changes in behaviour, morphology and colouration. The mechanisms behind these changes are unknown, as is whether the increased longevity is achieved through parasite manipulation. Here, we demonstrate that the parasite releases proteins into its host with functions that might explain the observed changes. These parasitic proteins make up a substantial portion of the proteome of the hosts' haemolymph, and thioredoxin peroxidase and superoxide dismutase, two antioxidants, exhibited the highest abundances among them. The largest part of the secreted proteins could not be annotated, indicating they are either novel or severely altered during recent coevolution to function in host manipulation. We also detected shifts in the hosts' proteome with infection, in particular an overabundance of vitellogenin‐like A in infected ants, a protein that regulates division of labour in Temnothorax ants, which could explain the observed behavioural changes. Our results thus suggest two different strategies that might be employed by this parasite to manipulate its host: secreting proteins with immediate influence on the host's phenotype and altering the host's translational activity. Our findings highlight the intricate molecular interplay required to influence the phenotype of a host and point to potential signalling pathways and genes involved in parasite–host communication. [ABSTRACT FROM AUTHOR]

Taxonomic evaluations are needed to accurately determine the host selection of fish parasites. The present study is a multidisciplinary research in the field of basic and fish diseases sciences. The description of the hybrid species of Squalius orientalis and Alburnus derjugini and infection of Ligula intestinalis in these hybrid fish were reported for the first time from the Kürtün Dam Lake in northeast Turkey. A total of 450 fish were sampled in March, August, and October in 2020 using gillnets. Detailed morphological characteristics (n = 24) were compared to determine the difference among ancestors and hybrid species. The prevalence of L. intestinalis between the sampling periods and the size groups of fish (0 – 10, 11 – 15, and ≥16 cm in length) were examined. Moreover, the highest prevalence of the parasite was observed in October (78.94 %), with a size range of 0 – 10 cm in length (77.8 %). In addition, the total prevalence of the parasite was 48.44 %. The results revealed that most of the diagnostic metric and meristic features of hybrid fish were ranging between the data of S. orientalis and A. derjugini. According to previous reports, when hybrid individuals were compared with their ancestors in terms of prevalence, hybrid individuals were more susceptible to L. intestinalis infections. This study was unique as it provided the first record of L. intestinalis in a hybrid fish population.

Copyright of Journal of Hydrobiology is the property of Editorial Department of Journal of Hydrobiology and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

This first comprehensive report from Punjab province of India relates to patho-physiological alterations alongwith morpho-molecular characterisation and risk assessment of natural infections of Hymenolepis diminuta and Hymenolepis nana in 291commensal rodents including house rat, Rattus rattus (n=201) and lesser bandicoot rat, Bandicota bangalensis (n=90). Small intestine of 53.61 and 64.95 % rats was found infected with H. diminuta and H. nana, respectively with a concurrent infection rate of 50.86 %. There was no association between male and female rats and H. diminuta and H. nana infections (ᵡ2 = 0.016 and 0.08, respectively, d.f.= 1, P>0.05), while the host age had significant effect on prevalence of H. diminuta and H. nana (ᵡ2 = 28.12 and 7.18, respectively, d.f.= 1, P≤0.05) infection. Examination of faecal samples and intestinal contents revealed globular shaped eggs of H. diminuta without polar filaments (76.50 ± 3.01μm x 67.62 ± 2.42 μm), while smaller sized oval eggs of H. nana were with 4 – 8 polar filaments (47.87 ± 1.95 μm x 36.12 ± 3.05 μm). Cestode infection caused enteritis, sloughing of intestinal mucosa, necrosis of villi and inflammatory reaction with infiltration of mononuclear cells in the mucosa and submucosa. Morphometric identification of the adult cestodes recovered from the intestinal lumen was confirmed by molecular characterisation based on nuclear ITS-2 loci which showed a single band of 269 bp and 242 bp for H. diminuta and H. nana, respectively. Pairwise alignment of the ITS-2 regions showed 99.46 % similarity with sequences of H. diminuta from USA and 100 % similarity with sequences of H. nana from Slovakia, Kosice.

A full account of the world's species is important for understanding how natural systems are structured, how they function and how they will respond to natural and anthropogenic pressures in the future. Despite the well-known importance of parasites in natural systems, they are too often underrepresented from studies on biodiversity and ecological functioning. In this review, we provide a quantitative synthesis of the current state of knowledge of helminth biodiversity within New Zealand's marine environment. We report that records of parasitic helminths within New Zealand marine animals are few and uneven across host taxa. A large proportion of parasite species are taxonomically unresolved, with some differences across parasite taxa. Few parasites have their whole life cycles resolved, and the majority are only reported from one host species at one life stage. Most endemic host species are yet to be investigated for helminths and there is likely an abundance of endemic parasite diversity yet to be discovered. Host species of greater conservation concern are being investigated for helminths less frequently than species of low conservation concern. We conclude that little is known about parasitic helminths in New Zealand's marine environment and make recommendations on how to improve this situation. [ABSTRACT FROM AUTHOR]

The first cestode genomes were obtained by an international consortium led by the Wellcome Sanger Institute that included representative institutions from countries where the sequenced parasites have been studied for decades, in part because they are etiological agents of endemic diseases (Argentina, Uruguay, Mexico, Canada, UK, Germany, Switzerland, Ireland, USA, Japan, and China). After this, several complete genomes were obtained reaching 16 species to date. Cestode genomes have smaller relative size compared to other animals including free-living flatworms. Moreover, the features genome size and repeat content seem to differ in the two analyzed orders. Cyclophyllidean species have smaller genomes and with fewer repetitive content than Diphyllobothriidean species. On average, cestode genomes have 13,753 genes with 6 exons per gene and 41% GC content. More than 5,000 shared cestode proteins were accurately annotated by the integration of gene predictions and transcriptome evidence being more than 40% of these proteins of unknown function. Several gene losses and reduction of gene families were found and could be related to the extreme parasitic lifestyle of these species. The application of cutting-edge sequencing technology allowed the characterization of the terminal sequences of chromosomes that possess unique characteristics. Here, we review the current status of knowledge of complete cestode genomes and place it within a comparative genomics perspective. Multidisciplinary work together with the implementation of new technologies will provide valuable information that can certainly improve our chances to finally eradicate or at least control diseases caused by cestodes. [ABSTRACT FROM AUTHOR]

A growing literature demonstrates the impact of helminths on their host gut microbiome. We investigated whether the stickleback host microbiome depends on ecoevolutionary variables by testing the impact of exposure to the cestode parasite Schistocephalus solidus with respect to infection success, host genotype, parasite genotype, and parasite microbiome composition. We observed constitutive differences in the microbiome of sticklebacks of different origin, and those differences increased when sticklebacks exposed to the parasite resisted infection. In contrast, the microbiome of successfully infected sticklebacks varied with parasite genotype. More specifically, we revealed that the association between microbiome and immune gene expression increased in infected individuals and varied with parasite genotype. In addition, we showed that S. solidus hosts a complex endomicrobiome and that bacterial abundance in the parasite correlates with expression of host immune genes. Within this comprehensive analysis we demonstrated that (i) parasites contribute to modulating the host microbiome through both successful and unsuccessful infection, (ii) when infection is successful, the host microbiome varies with parasite genotype due to genotype‐dependent variation in parasite immunomodulation, and (iii) the parasite‐associated microbiome is distinct from its host and impacts the host immune response to infection. [ABSTRACT FROM AUTHOR]

Alveolar echinococcosis, the disease caused by infection with the intermediate stage of the Echinococcus multilocularis tapeworm, is typically fatal in humans and dogs when left untreated. Since 2012, alveolar echinococcosis has been diagnosed in 5 dogs, 3 lemurs, and 1 chipmunk in southern Ontario, Canada, a region previously considered free of these tapeworms. Because of human and animal health concerns, we estimated prevalence of infection in wild canids across southern Ontario. During 2015–2017, we collected fecal samples from 460 wild canids (416 coyotes, 44 foxes) during postmortem examination and analyzed them by using a semiautomated magnetic capture probe DNA extraction and real-time PCR method for E. multilocularis DNA. Surprisingly, 23% (95% CI 20%–27%) of samples tested positive. By using a spatial scan test, we identified an infection cluster (relative risk 2.26; p = 0.002) in the western-central region of the province. The cluster encompasses areas of dense human population, suggesting zoonotic transmission.

Gastrointestinal helminth parasites are a concern for the poultry industry worldwide as they can affect the health, welfare, and production performance. A systematic review of the prevalence over time in different countries may improve our understanding of gastrointestinal helminthiasis in chickens and subsequently lead to improved poultry health. The aim of this systematic review and meta-analysis was to provide an overview of the published information regarding the epidemiology and the diagnostic approaches of chicken helminth infection. Six databases were searched for studies, and a total of 2,985 articles published between 1942 and 2019 were identified and subsequently screened for eligibility using title or abstract and full text assessment, resulting in 191 publications to be used in the study. Postmortem diagnostics (73.8%) and the flotation technique (28.8%) were commonly used to detect helminth infections with a pooled prevalence of 79.4% ranging from 4 to 100%. More than 30 helminth species in chicken populations were identified including Ascaridia galli (35.9%), Heterakis gallinarum (28.5%), Capillaria spp. (5.90%), and Raillietina spp. (19.0%) being the most prevalent. The reported prevalence of helminth infection decreased over time in developing countries while it increased in the developed world. Chicken kept in backyard and free-range systems had a markedly higher pooled prevalence of helminth infection (82.6 and 84.8%, respectively) than those housed in cage production systems (63.6%). This may indicate the need for more rigorous control and prevention measures in free-range and backyard production systems using regular deworming coupled with access to early and accurate diagnosis allowing for early intervention.

A patient in Pennsylvania, USA, with common variable immunodeficiency sought care for fever, cough, and abdominal pain. Imaging revealed lesions involving multiple organs. Liver resection demonstrated necrotizing granulomas, recognizable tegument, and calcareous corpuscles indicative of an invasive cestode infection. Sequencing revealed 98% identity to a Versteria species of cestode found in mink.

This paper provides the first report of the invasive Asian fish tapeworm, Schyzocotyle (Bothriocephalus) acheilognathi (Yamaguti, 1934), from Norway and Scandinavia. The parasite was found during routine post mortem disease examination of an imported koi carp, Cyprinus carpio, from an outdoor pond in the North of Norway. The intestine contained live tapeworms and these tapeworms were identified as S. acheilognathi by DNA sequencing of partial 18S and 28S ribosomal DNA and comparison of the obtained sequences with sequences in GenBank. The infected fish specimen along with the other fish in the pond were exterminated. The risk of infection from the pond to other fish outside of the pond was thus negligible. However, the finding of S. acheilognathi in imported aquarium fish shows that such import poses a risk of introducing pathogens to new areas. Had this fish pond been situated in the south of Norway where the temperatures are higher and where susceptible hosts are readily available, an escape of fish or release of eggs from the pond could potentially have resulted in infection of local fish populations. [ABSTRACT FROM AUTHOR]

Cephalopods are intermediate/paratenic hosts in the life cycle of elasmobranch tapeworms, nevertheless most records of infection in this group of mollusks are outdated and fragmentary. The present work aimed to investigate the cestode fauna of the common octopus Octopus vulgaris from the Tyrrhenian Sea (Central Mediterranean). The parasitic stages were characterized by light and Scanning Electron Microscopy (SEM) and sequencing of 28S rDNA. Three cestode taxa were identified to the genus level: the onchoproteocephalidean Acanthobothrium sp. (prevalence 28%), the “tetraphyllidean” Anthobothrium sp. (prevalence 13%) and the trypanorhynch Nybelinia sp. (prevalence 3%). The remarkable prevalence observed for gastrointestinal cestodes highlight a possible important role of O. vulgaris in the transmission of elasmobranch tapeworms, particularly Onchoproteocephalideans. Furthermore, the present work provides, for the first time, detailed morphological (SEM) and molecular support to confirm the occurrence of Anthobothrium sp. in cephalopod hosts. In order to gain higher taxonomic resolution for the identified taxa, we stress the need to collect further morphological and molecular data of adult cestodes infecting their elasmobranch definitive hosts.

Abstract Background microRNAs (miRNAs), a class of small non-coding RNAs, are key regulators of gene expression at post-transcriptional level and play essential roles in fundamental biological processes such as development and metabolism. The particular developmental and metabolic characteristics of cestode parasites highlight the importance of studying miRNA gene regulation in these organisms. Here, we perform a comprehensive analysis of miRNAs in the parasitic cestode Echinococcus canadensis G7, one of the causative agents of the neglected zoonotic disease cystic echinococcosis. Methods Small RNA libraries from protoscoleces and cyst walls of E. canadensis G7 and protoscoleces of E. granulosus sensu stricto G1 were sequenced using Illumina technology. For miRNA prediction, miRDeep2 core algorithm was used. The output list of candidate precursors was manually curated to generate a high confidence set of miRNAs. Differential expression analysis of miRNAs between stages or species was estimated with DESeq. Expression levels of selected miRNAs were validated using poly-A RT-qPCR. Results In this study we used a high-throughput approach and found transcriptional evidence of 37 miRNAs thus expanding the miRNA repertoire of E. canadensis G7. Differential expression analysis showed highly regulated miRNAs between life cycle stages, suggesting a role in maintaining the features of each developmental stage or in the regulation of developmental timing. In this work we characterize conserved and novel Echinococcus miRNAs which represent 30 unique miRNA families. Here we confirmed the remarkable loss of conserved miRNA families in E. canadensis, reflecting their low morphological complexity and high adaptation to parasitism. Conclusions We performed the first in-depth study profiling of small RNAs in the zoonotic parasite E. canadensis G7. We found that miRNAs are the preponderant small RNA silencing molecules, suggesting that these small RNAs could be an essential mechanism of gene regulation in this species. We also identified both parasite specific and divergent miRNAs which are potential biomarkers of infection. This study will provide valuable information for better understanding of the complex biology of this parasite and could help to find new potential targets for therapy and/or diagnosis.

Mesocestoides corti is a widely used model for the study of cestode biology, and its transition from the larval tetrathyridium (TT) stage to the strobilated, adult worm (ST) stage can be induced and followed in vitro . Here, a proteomic approach was used to describe and compare M. corti TT and ST protein repertories. Overall, 571 proteins were identified, 238 proteins in TT samples and 333 proteins in ST samples. Among the identified proteins, 207 proteins were shared by TTs and STs, while 157 were stage-specific, being 31 exclusive from TTs, and 126 from STs. Functional annotation revealed fundamental metabolic differences between the TT and the ST stages. TTs perform functions related mainly to basic metabolism, responsible for growth and vegetative development by asexual reproduction. STs, in contrast, perform a wider range of functions, including macromolecule biosynthetic processes, gene expression and control pathways, which may be associated to its proglottization/segmentation, sexual differentiation and more complex physiology. Furthermore, the generated results provided an extensive list of cestode proteins of interest for functional studies in M. corti . Many of these proteins are novel candidate diagnostic antigens, and/or potential targets for the development of new and more effective antihelminthic drugs. Biological significance Cestodiases are parasitic diseases with serious impact on human and animal health. Efforts to develop more effective strategies for diagnosis, treatment or control of cestodiases are impaired by the still limited knowledge on many aspects of cestode biology, including the complex developmental processes that occur in the life cycles of these parasites. Mesocestoides corti is a good experimental model to study the transition from the larval to the adult stage, called strobilation, which occur in typical cestode life-cycles. The performed proteomics approach provided large-scale identification and quantification of M. corti proteins. Many stage-specific or differentially expressed proteins were detected in the larval tetrathyridium (TT) stage and in the strobilated, adult worm (ST) stage. Functional comparative analyses of the described protein repertoires shed light on function and processes associated to specific features of both stages, such as less differentiation and asexual reproduction in TTs, and proglottization/segmentation and sexual differentiation in ST. Moreover, many of the identified stage-specific proteins are useful as cestode developmental markers, and are potential targets for development of novel diagnostic methods and therapeutic drugs for cestodiases. [ABSTRACT FROM AUTHOR]

Two digging decapod crustaceans, the sand crab species Lepidopa benedicti and the mole crab species Emerita benedicti, both live in the swash zone of fine sand beaches. They were examined for two parasites that infect decapod crustaceans in the region, an unidentified nematode previously shown to infect L. benedicti, and cestode tapeworm larvae, Polypocephalus sp., previously shown to infect shrimp (Litopenaeus setiferus). Lepidopa benedicti were almost always infected with both parasite species, while E. benedicti were rarely infected with either parasite species. This difference in infection pattern suggests that tapeworms are ingested during sediment feeding in L. benedicti, which E. benedicti avoid by filter feeding. Larger L. benedicti had more Polypocephalus sp. larvae. The thoracic ganglia, which make up the largest proportion of neural tissue, contained the largest numbers of Polypocephalus sp. larvae. Intensity of Polypocephalus sp. infection was not correlated with how long L. benedicti remained above sand in behavioural tests, suggesting that Polypocephalus sp. do not manipulate the sand crabs in a way that facilitates trophic transmission of the parasite. Litopenaeus setiferus may be a primary host for Polypocephalus sp., and L. benedict may be a secondary, auxiliary host.

Echinococcus equinus causes cystic echinococcosis in equines. The cestode appears to be endemic in Spain, Italy, Great Britain and Ireland. The present case shows the emergence of the parasite to our previously unaffected latitude. A 17-year old Connemara horse, born and raised in Ireland, was brought to Germany at the age of 10 years. Here, it developed an orosinuidal fistula and chronic colitis and was euthanized with poor prognosis. Necropsy revealed multiple well-demarcated, thick-walled, grey-white hydatid cysts of 1 to 5cm in diameter in the liver and lung. PCR product sequencing revealed complete identity with E. equinus. The present case indicates the spreading of an infectious agent outside its original endemic region. In order to prevent further spreading of Echinococcus equinus special attention should be given to the import of dogs and horses from endemic regions as well as to dogs being fed with raw horseorgans or contaminated meat in terms of the Biologically Appropriate Raw Foods (BARF) –diet which has gained increasing popularity. These dogs may potentially become reservoirs for Echinococcus equinus, a risk which should be considered. [ABSTRACT FROM AUTHOR]

Parasites with complex life cycles have developed numerous phenotypic strategies, closely associated with developmental events, to enable the exploitation of different ecological niches and facilitate transmission between hosts. How these environmental shifts are regulated from a metabolic and physiological standpoint, however, still remain to be fully elucidated. We examined the transcriptomic response of Schistocephalus solidus, a trophically transmitted parasite with a complex life cycle, over the course of its development in an intermediate host, the threespine stickleback, and the final avian host. Results from our differential gene expression analysis show major reprogramming events among developmental stages. The final host stage is characterized by a strong activation of reproductive pathways and redox homoeostasis. The attainment of infectivity in the fish intermediate host-which precedes sexual maturation in the final host and is associated with host behaviour changes-is marked by transcription of genes involved in neural pathways and sensory perception. Our results suggest that un-annotated and S. solidus-specific genes could play a determinant role in host-parasite molecular interactions required to complete the parasite's life cycle. Our results permit future comparative analyses to help disentangle species-specific patterns of infection from conserved mechanisms, ultimately leading to a better understanding of the molecular control and evolution of complex life cycles. [ABSTRACT FROM AUTHOR]

A patient in Pennsylvania, USA, with common variable immunodeficiency sought care for fever, cough, and abdominal pain. Imaging revealed lesions involving multiple organs. Liver resection demonstrated necrotizing granulomas, recognizable tegument, and calcareous corpuscles indicative of an invasive cestode infection. Sequencing revealed 98% identity to a Versteria species of cestode found in mink. [ABSTRACT FROM AUTHOR]

A patient in Pennsylvania, USA, with common variable immunodeficiency sought care for fever, cough, and abdominal pain. Imaging revealed lesions involving multiple organs. Liver resection demonstrated necrotizing granulomas, recognizable tegument, and calcareous corpuscles indicative of an invasive cestode infection. Sequencing revealed 98% identity to a Versteria species of cestode found in mink. [ABSTRACT FROM AUTHOR]

Parasitic helminths display highly complex life-cycles in which the establishment of adults or larvae within host target organs as well as the transition of one developmental stage to the following is influenced by host-derived factors. Due to its approachability concerning in vitro cultivation, the larval stage of the fox-tapeworm Echinococcus multilocularis has recently emerged as a model system to study the molecular nature of such host-derived stimuli and their influence on developmental control in the parasite. Data obtained so far indicate that cytokines which are used by the host for cell-cell communication can also be exploited by the parasite as clues to find suitable target organs. This involves direct interactions of evolutionary conserved signalling systems of the receptor tyrosine – and the receptor serine/threonine-kinase pathways of the parasite with corresponding host cytokines of the insulin –, the epidermal growth factor-, and the transforming growth factor-β-families. In the present article, we will briefly review in vitro cultivation approaches undertaken so far for E. multilocularis larvae as well as our current knowledge on the parasite’s signalling systems and their interaction with host cytokines.

Background: Schistocephalus solidus is a well-established model organism for studying the complex life cycle of cestodes and the mechanisms underlying host-parasite interactions. However, very few large-scale genetic resources for this species are available. We have sequenced and de novo-assembled the transcriptome of S. solidus using tissues from whole worms at three key developmental states - non-infective plerocercoid, infective plerocercoid and adult plerocercoid - to provide a resource for studying the evolution of complex life cycles and, more specifically, how parasites modulate their interactions with their hosts during development. Findings: The de novo transcriptome assembly reconstructed the coding sequence of 10,285 high-confidence unigenes from which 24,765 non-redundant transcripts were derived. 7,920 (77 %) of these unigenes were annotated with a protein name and 7,323 (71 %) were assigned at least one Gene Ontology term. Our raw transcriptome assembly (unfiltered transcripts) covers 92 % of the predicted transcriptome derived from the S. solidus draft genome assembly currently available on WormBase. It also provides new ecological information and orthology relationships to further annotate the current WormBase transcriptome and genome. Conclusion: This large-scale transcriptomic dataset provides a foundation for studies on how parasitic species with complex life cycles modulate their response to changes in biotic and abiotic conditions experienced inside their various hosts, which is a fundamental objective of parasitology. Furthermore, this resource will help in the validation of the S solidus gene features that have been predicted based on genomic sequence. [ABSTRACT FROM AUTHOR]

Background: Schistocephalus solidus is a well-established model organism for studying the complex life cycle of cestodes and the mechanisms underlying host-parasite interactions. However, very few large-scale genetic resources for this species are available. We have sequenced and de novo-assembled the transcriptome of S. solidus using tissues from whole worms at three key developmental states - non-infective plerocercoid, infective plerocercoid and adult plerocercoid - to provide a resource for studying the evolution of complex life cycles and, more specifically, how parasites modulate their interactions with their hosts during development. Findings: The de novo transcriptome assembly reconstructed the coding sequence of 10,285 high-confidence unigenes from which 24,765 non-redundant transcripts were derived. 7,920 (77%) of these unigenes were annotated with a protein name and 7,323 (71%) were assigned at least one Gene Ontology term. Our raw transcriptome assembly (unfiltered transcripts) covers 92% of the predicted transcriptome derived from the S. solidus draft genome assembly currently available on WormBase. It also provides new ecological information and orthology relationships to further annotate the current WormBase transcriptome and genome. Conclusion: This large-scale transcriptomic dataset provides a foundation for studies on how parasitic species with complex life cycles modulate their response to changes in biotic and abiotic conditions experienced inside their various hosts, which is a fundamental objective of parasitology. Furthermore, this resource will help in the validation of the S solidus gene features that have been predicted based on genomic sequence. [ABSTRACT FROM AUTHOR]

Fish constitute a major component of diet for the people of Northeast India and they are extensively used as a protein-rich food for human consumption. The present studies incorporate the spectrum, composition and diversity of the parasitic species in freshwater fishes in Meghalaya, Northeast India, with a view to identifying the species recovered by morphological criteria based on light microscopy. The collection sites included sixteen foci from eleven districts of Meghalaya for parasites occurring in the common food fishes (Cypriniformes, Channiformes, Silurformes, Symbranchiformes and Anguilliformes). The helminth parasite spectrum recovered from the various piscine host species in the study area comprised of a total of 19 taxa: 2 monogenean, 8 trematode (4 adult and 4 metacercarial stages), 12 cestode (11 adult and a metacestode stage), 6 nematode (3 adult and 3 larval stages) and a single acanthocephalan species. A checklist of the parasite species with short remarks for each is provided herein. [ABSTRACT FROM AUTHOR]

Background: microRNAs (miRNAs), a class of small non-coding RNAs, are key regulators of gene expression at post-transcriptional level and play essential roles in fundamental biological processes such as development and metabolism. The particular developmental and metabolic characteristics of cestode parasites highlight the importance of studying miRNA gene regulation in these organisms. Here, we perform a comprehensive analysis of miRNAs in the parasitic cestode Echinococcus canadensis G7, one of the causative agents of the neglected zoonotic disease cystic echinococcosis. Methods: Small RNA libraries from protoscoleces and cyst walls of E. canadensis G7 and protoscoleces of E. granulosus sensu stricto G1 were sequenced using Illumina technology. For miRNA prediction, miRDeep2 core algorithm was used. The output list of candidate precursors was manually curated to generate a high confidence set of miRNAs. Differential expression analysis of miRNAs between stages or species was estimated with DESeq. Expression levels of selected miRNAs were validated using poly-A RT-qPCR. Results: In this study we used a high-throughput approach and found transcriptional evidence of 37 miRNAs thus expanding the miRNA repertoire of E. canadensis G7. Differential expression analysis showed highly regulated miRNAs between life cycle stages, suggesting a role in maintaining the features of each developmental stage or in the regulation of developmental timing. In this work we characterize conserved and novel Echinococcus miRNAs which represent 30 unique miRNA families. Here we confirmed the remarkable loss of conserved miRNA families in E. canadensis, reflecting their low morphological complexity and high adaptation to parasitism. Conclusions: We performed the first in-depth study profiling of small RNAs in the zoonotic parasite E. canadensis G7. We found that miRNAs are the preponderant small RNA silencing molecules, suggesting that these small RNAs could be an essential mechanism of gene regulation in this species. We also identified both parasite specific and divergent miRNAs which are potential biomarkers of infection. This study will provide valuable information for better understanding of the complex biology of this parasite and could help to find new potential targets for therapy and/or diagnosis. [ABSTRACT FROM AUTHOR]

Highlights: [•] Helminth germinal cells constitute a highly relevant tool to study the developmental processes. [•] The establishment of a cell line from the germinal layer was achieved. [•] Monolayer cells survived cryopreservation and were able to proliferate after thawing. [•] Attached cells showed a high degree of heteroploidy. [•] The cell line could be useful for practical purposes including drug screening. [ABSTRACT FROM AUTHOR]

Abstract: In this study we demonstrate RNA interference mediated knock-down of target gene expression in Echinococcus multilocularis primary cells on both the transcriptional and translational level. In addition, we report on an improved method for generating E. multilocularis primary cell mini-aggregates from in vitro cultivated metacestode vesicles, and on the cultivation of small numbers of small interfering RNA-transfected cells in vitro over an extended period of time. This allows assessments on the effects of RNA interference performed on Echinococcus primary cells with regard to growth, proliferation, differentiation of the parasite and the formation of novel metacestode vesicles in vitro. [ABSTRACT FROM AUTHOR]

Abstract: Cell cultures of parasitic helminths are an invaluable tool for investigations of basic biological processes, as well as for development of improved chemotherapeutic agents and molecular interactions between host and parasite. We carried out a simple and efficient methodology to isolate Echinococcus granulosus germinal cells which were maintained for at least 4months while cultivated in the presence of reducing agents and hormones. Microscopic analysis of the primary cell culture revealed the presence of cells with similar Echinococcus germinal cell morphology and behaviour. Population doubling time was estimated at 48h, showing a rapid division rate. To discard possible host contamination, the specificity of the primary culture was tested by nested PCR, analyzing mdh gene expression and obtaining only one product with the expected size. We also studied the expression of specific E. granulosus proteins in primary cell culture. The novel and systematized method described here constitutes a powerful tool for investigations in cystic echinococcosis on biochemical and biological aspects related to the life cycle of the parasite and mechanisms of host–parasite interactions. This method also constitutes a powerful tool for the design of more efficient therapeutic alternatives. [Copyright &y& Elsevier]

Abstract: Mitogen-activated protein kinase kinases (MAPKKs) are essential components of evolutionary conserved signalling modules that regulate a variety of fundamental cellular processes in response to environmental stimuli. To date, no MAPKK ortholog has been characterised in free-living or parasitic flatworm species. Here, we report the identification and molecular characterisation of two such molecules in the human parasitic cestode Echinococcus multilocularis, the causative agent of alveolar echinococcosis. Using degenerative PCR approaches as well as 3′- and 5′-rapid amplification of cDNA ends (RACE), the cDNAs encoding two different E. multilocularis MAPKKs, EmMKK1 and EmMKK2, have been identified and fully cloned. Structurally, EmMKK1 and EmMKK2 closely resemble members of the MKK3/6- and the MEK1/2-MAPKK sub-families, respectively, from a variety of vertebrate and invertebrate organisms, and contain all catalytically important residues of MAPKKs at the corresponding positions. By reverse transcriptase-PCR analyses, expression of the EmMKK2-encoding gene, emmkk2, was observed in the larval stages, metacestode and protoscolex while emmkk1 displayed a protoscolex-specific expression pattern. In yeast two-hybrid analyses, EmMKK1 strongly interacted with the previously identified Echinococcus MAPKK kinase EmRaf but not with the Erk-like MAP kinase EmMPK1 or the p38-like MAP kinase EmMPK2. EmMKK2, on the other hand, not only interacted with EmRaf and a member of the parasite’s 14-3-3 protein family, but also with EmMPK1, which was confirmed by co-immunoprecipitation assays. Incubation of in vitro cultivated metacestode vesicles with small-molecule inhibitors of Raf- and MEK-kinases resulted in a marked de-phosphorylation of EmMPK1 and negatively affected parasite growth, but was ineffective in vesicle killing. Taken together, our results define EmRaf, EmMKK2 and EmMPK1 as the three components of the Erk-like E. multilocularis MAPK cascade module and provide a solid basis for further investigations into the role of Erk-like MAPK signalling in parasite development and stem cell function. [Copyright &y& Elsevier]

Ultrastructure studies of internuncial processes (IP) in the tegument for different body zones in a cyclophyllid davaineid adult cestode, Raillietina (Fuhrmannetta) echinobothrida infecting the small intestine of country fowl, Gallus domesticus in Naihati area of West Bengal, India have been investigated. Transmission electron microscopic observation reveals that the type, distribution, diameter, and overall characteristics of the IP along the zones of the strobila differ greatly. Contrary to the existing findings the IPs in this cestode have been recorded as thin and broad type. Thin type IP is common in all the body zones except the mature proglottides. Broad type IP is not found in the gravid proglottides. This type of IP is elaborately established in the mature proglottides. Development of broad type IP has been studied for the first time. In the gravid proglottides, IPs are either in receding nature or altogether absent. The genesis of these two types of IPs has been put forward and their functions are assigned. [ABSTRACT FROM AUTHOR]

Abstract: The tegumental membrane of Taenia crassiceps cysticerci contains an ATP-diphosphohydrolase (EC 3.6.1.5) which hydrolyzes purine and pyrimidine nucleoside 5′-di- and 5′-triphosphates at an optimum pH of 8.5. It is Mg2+-dependent and insensitive to classical ATPase and phosphatase inhibitors. In solubilized tegumental membrane the Km values varied from 220 to 480μM and the V max from 370 to 748nmol of Pi release/mg/min for nucleoside triphosphates (ATP, GTP, CTP, UTP, and TTP); for nucleoside diphosphates (ADP, GDP, CDP, and UDP) the Km values were from 260 to 450μM and the V max from 628 to 1134nmol of Pi release/mg/min. An antibody specific to CD39 shows cross-reactivity with T. crassiceps ATP-diphosphohydrolase, revealing a single protein of ∼80kDa. Incubation of ATP-diphosphohydrolase with FSBA inhibited ATPase and ADPase activities by 85–90%. Immunoblot analyses, the competition plot, similar inhibition by free nucleotides, the lack of effect of Mg2+ at high concentrations, and the inactivation by FSBA of ATPase and ADPase activity strongly suggest that a single enzyme catalyzes the hydrolysis of all these nucleotides. The mechanism of ATP hydrolysis shows that ATP-diphosphohydrolase releases ADP during the catalytic cycle. Incubation of intact cysticerci with FSBA caused 70–80% inhibition of ATPase and ADPase activities, indicating that the active site of the ATP-diphosphohydrolase is oriented to the external surface of the tegument of T. crassiceps. The importance of this enzyme in the parasite–host relationship is discussed. [Copyright &y& Elsevier]

Abstract: Alveolar echinococcosis (AE), caused by the metacestode larval stage of the fox-tapeworm Echinococcus multilocularis, is a life-threatening disease with very limited treatment options. In search for novel drug targets, we concentrate on factors of the cellular signaling machinery and report herein the characterization of a novel gene, Emmpk2, which is expressed in the parasite’s larval stage and which codes for a member of the mitogen-activated protein kinase (MAPK) family. On the amino acid sequence level, the encoded protein, EmMPK2, shares considerable homologies with p38 MAPKs from a wide variety of animal organisms but also displays several distinct differences, particularly in amino acid residues known to be involved in the regulation of enzyme activity. Upon heterologous expression in Escherichia coli, purified EmMPK2 showed prominent autophosphorylation activity and strongly elevated basal activity towards a MAPK substrate, when compared to the closest human orthologue, p38-α. EmMPK2 activity could be effectively inhibited in the presence of ML3403 and SB202190, two ATP-competitive pyridinyl imidazole inhibitors of p38 MAPKs, in a concentration-dependent manner. When added to in vitro cultivated metacestode vesicles, SB202190 and particularly ML3403 led to dephosphorylation of EmMPK2 in the parasite and effectively killed parasite vesicles at concentrations that did not affect cultivated mammalian cells. Taken together, these results identify pyridinyl imidazoles as a novel class of anti-Echinococcus compounds and EmMPK2 as a promising target for the development of drugs against alveolar echinococcosis. [Copyright &y& Elsevier]

Abstract: A major limitation in studying molecular interactions between parasitic helminths and their hosts is the lack of suitable in vitro cultivation systems for helminth cells and larvae. Here we present a method for long-term in vitro cultivation of larval cells of the tapeworm Echinococcus multilocularis, the causative agent of alveolar echinococcosis. Primary cells isolated from cultivated metacestode vesicles in vitro showed a morphology typical of Echinococcus germinal cells, displayed an Echinococcus-specific gene expression profile and a cestode-like DNA content of ∼300Mbp. When kept under reducing conditions in the presence of Echinococcus vesicle fluid, the primary cells could be maintained in vitro for several months and proliferated. Most interestingly, upon co-cultivation with host hepatocytes in a trans-well system, mitotically active Echinococcus cells formed cell aggregates that subsequently developed central cavities, surrounded by germinal cells. After 4 weeks, the cell aggregates gave rise to young metacestode vesicles lacking an outer laminated layer. This layer was formed after 6 weeks of cultivation indicating the complete in vitro regeneration of metacestode larvae. As an initial step toward the creation of a fully transgenic strain, we carried out transient transfection of Echinococcus primary cells using plasmids and obtained heterologous expression of a reporter gene. Furthermore, we successfully carried out targeted infection of Echinococcus cells with the facultatively intracellular bacterium Listeria monocytogenes, a DNA delivery system for genetic manipulation of mammalian cells. Taken together, the methods presented herein constitute important new tools for molecular investigations on host–parasite interactions in alveolar echinococcosis and on the roles of totipotent germinal cells in parasite regeneration and metastasis formation. Moreover, they enable the development of fully transgenic techniques in this group of helminth parasites for the first time. [Copyright &y& Elsevier]

Abstract: Members of the transforming growth factor-β/bone morphogenetic protein (TGF-β/BMP) family of cytokines play crucial roles in animal development and are candidate molecules for host–parasite cross-communication in helminth diseases. TGF-β/BMP-signalling involves binding of the cytokines to receptor kinases which subsequently activate intracellular transcription factors of the Smad family. We have previously characterized two members of the receptor-regulated Smad (R-Smad) family, EmSmadA and EmSmadB, from the human parasitic cestode Echinococcus multilocularis and now present evidence for two additional Smads that are expressed by the larval stages of the parasite. The full-length cDNAs coding for a third R-Smad, EmSmadC, and a common mediator Smad (Co-Smad), EmSmadD, were characterized. While EmSmadD displayed a typical Co-Smad structure, EmSmadC lacked the N-terminal MH1 domain which is typically found in Smads. In yeast two-hybrid analyses, EmSmadC and EmSmadD were capable of homo- and heterodimer formation with other Echinococcus Smads. Furthermore, EmSmadC displayed autonomous transcription activation activity and interacted with EmSkip, a member of the SNW/SKIP family of transcriptional regulators. In a heterologous expression system, EmSmadC was specifically phosphorylated by mammalian TGF-β receptors, indicating that it is a member of the AR-Smad sub-family. Finally, in activity assays, the parasite’s Erk-like kinase EmMPK1 phosphorylated EmSmadD, indicating cross-regulation between mitogen-activated protein kinase cascade- and TGF-β/BMP-signalling in Echinococcus. The data presented herein significantly broaden our knowledge of Smad-signalling factors in E. multilocularis and will facilitate studies on TGF-β/BMP-regulated genes in the parasite as well as TGF-β/BMP mediated host–parasite cross-interaction during alveolar echinococcosis. [Copyright &y& Elsevier]

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Abstract: Mitogen-activated protein (MAP) kinases are key regulators of cellular signalling systems that mediate responses to a wide variety of extracellular stimuli and should also play a central role in developmental mechanisms of parasitic helminths. Until now, however, no MAP kinase orthologue has been characterised in a member of this parasite group. Here, we report the identification and characterisation of such a molecule, EmMPK1, from the human parasitic cestode Echinococcus multilocularis. Using a degenerative PCR approach, we isolated and completely sequenced the 1.2kb cDNA for EmMPK1 which displays significant homologies to known MAP kinases of different phylogenetic origin. EmMPK1 contains all amino acid residues which are characteristic for MAP kinases, including a conserved TEY motif which identifies the protein as a member of the ERK subfamily of MAP kinases. The corresponding gene, emmpk1 (6.9kb), was characterised and contained 10 introns. Southern blot hybridisation studies showed that emmpk1 is present as single copy locus in E. multilocularis. Using RT-PCR analyses we demonstrated that emmpk1 is expressed in form of three different transcripts which derive from alternative splice acceptor site utilisation at intron 9. Using EmMPK1-specific antibodies in Western blot studies and immunohistochemistry, we detected the Echinococcus protein and its phosphorylated form in the larval stages metacestode and protoscolex during in vitro cultivation and during an infection of the intermediate host. EmMPK1, immunoprecipitated from Echinococcus lysate, was able to phosphorylate myelin basic protein in activity assays, indicating that it is a functionally active MAP kinase. Finally, we also show that phosphorylation of EmMPK1 is specifically induced in vitro-cultivated E. multilocularis metacestode vesicles in response to exogenous host serum and upon addition of human epidermal growth factor. These data indicate that the E. multilocularis metacestode is able to sense epidermal growth factor from the host which results in an activation of the parasite’s MAP kinase cascade. [Copyright &y& Elsevier]

Abstract: Recombinant antigens that have been cloned from Taenia solium and Taenia ovis have been shown to be highly effective when used as vaccines against cysticercosis in the intermediate hosts. This study investigated the presence of mRNA encoding the TSOL18 and TSOL16 antigens in different life-cycle stages of T. solium, and their related homologues in T. ovis. Reverse transcription-PCR and Southern blotting demonstrated that the antigens are stage-specifically expressed in the oncosphere. The apparent absence of expression of TSOL18 in the metacestode life-cycle stage suggests that the vaccine based on this antigen targets exclusively the early stages in the development of the parasite. [Copyright &y& Elsevier]

Abstract: The alcoholic crude root-peel extract of Flemingia vestita and its major isoflavone, genistein, have been shown to have a vermifugal/vermicidal effect by causing a flaccid paralysis accompanied by alterations in the structural architecture of the tegumental interface and metabolic activity in Raillietina echinobothrida, the cestode of domestic fowl. In the present study, the crude root-peel extract and pure genistein were tested in vitro with respect to Ca2+ homeostasis and the occurrence of some metal ions was detected in the parasite. Live cestodes were incubated in pre-defined concentrations of the crude root-peel extract, genistein and praziquantel (as reference drug), till the paralysis time with simultaneous maintenance of respective controls. In the parasite tissue, a significant amount of Ca2+ (∼400 μg/g dry tissue wt) was found to be present besides magnesium, iron, zinc, lead and chromium, whilst manganese, cadmium and nickel were below the level of detection. The Ca2+ concentration was decreased significantly by 39%–49%, in the parasite tissue exposed to the test materials in comparison to the respective controls. There was also an increase in Ca2+ efflux by 91%–160% into the culture medium under similar treatments. The changes in Ca2+ homeostasis may be related to the rapid muscular contraction and consequent paralysis in the parasite due to the anthelmintic stress caused by the phytochemicals of F. vestita. [Copyright &y& Elsevier]