Wood mouse (Apodemus sylvaticus L.) as intermediate host for Mesocestoides canislagopodis (Rudolphi, 1810) (Krabbe 1865) in Iceland.

Mesocestoides canislagopodis is a common parasite of the arctic fox (Vulpes lagopus) in Iceland. In the past, household dogs (Canis familiaris) and cats (Felis catus) were also reported in Iceland to be infected. Recently, scolices of a non-maturing Mesocestoides sp. were detected in the intestines of the gyrfalcon (Falco rusticolus), and tetrathyridia were isolated in the body cavity of rock ptarmigan (Lagopus muta) and subsequently described. All stages were confirmed, using both morphological and molecular methods, to belong to the same species, M. canislagopodis. In the present study, post-mortem examination of wood mice (Apodemus sylvaticus), sampled in autumn 2014 on a farm in Northeast Iceland, revealed the presence of tetrathyridia in the peritoneal cavity and in the liver. Most tetrathyridia in the peritoneal cavity were free, but some were encapsulated in a thin connective tissue stroma and loosely attached to the inner organs. They appear as whitish, heart-shaped, flattened, unsegmented bodies with a slightly pointed posterior end. In the liver, tetrathyridia were seen as pale-tanned nodules embedded in the parenchyma. Comparative molecular analysis, both at the generic level (D1 domain LSU ribosomal DNA), and at the specific level (cytochrome c oxidase subunit I (cox1) and 12S mitochondrial DNA), revealed that the tetrathyridia belonged to M. canislagopodis. A. sylvaticus represents a new second intermediate host record in Iceland, and the first description of a rodent as intermediate host for this species, thus participating in the life cycle of the parasite.

Tegument Ultrastructure and Morphology of the Capsule Surrounding the Tetrathyridia of the Genus Mesocestoides Vaillant, 1863 in the Liver of the Root Vole.

The ultrastructure of the tegument of encapsulated tetrathyridia of the genus Mesocestoides Vaillant, 1863 (Cestoda, Cyclophyllidea, Mesocestoididae) from the liver of root voles Microtus oeconomus (Pallas, 1776) and the structure of the three-layered capsule surrounding them were studied for the first time. Several types of extracellular structures were noted on the surface of the tetrathyridia tegument: vesicles, fine granular material, and vacuoles. In addition, the phenomenon of shedding microtriches, which have expanded parts, was found. Host cells in contact with extracellular material show signs of destruction. A characteristic feature of the capsules surrounding the tetrathyridia is the reticular structure of the fibrous layer containing both native and degenerating inflammatory cells.

Cell repertoire and proliferation of germinative cells of the model cestode Mesocestoides corti.

The phylum Platyhelminthes shares a unique population of undifferentiated cells responsible for the proliferation capacity needed for cell renewal, growth, tissue repair and regeneration. These cells have been extensively studied in free-living flatworms, whereas in cestodes the presence of a set of undifferentiated cells, known as germinative cells, has been demonstrated in classical morphology studies, but poorly characterized with molecular biology approaches. Furthermore, several genes have been identified as neoblast markers in free-living flatworms that deserve study in cestode models. Here, different cell types of the model cestode Mesocestoides corti were characterized, identifying differentiated and germinative cells. Muscle cells, tegumental cells, calcareous corpuscle precursor cells and excretory system cells were identified, all of which are non-proliferative, differentiated cell types. Besides those, germinative cells were identified as a population of small cells with proliferative capacity in vivo. Primary cell culture experiments in Dulbecco's Modified Eagle Medium (DMEM), Echinococcus hydatid fluid and hepatocyte conditioned media in non-reductive or reductive conditions confirmed that the germinative cells were the only ones with proliferative capacity. Since several genes have been identified as markers of undifferentiated neoblast cells in free-living flatworms, the expression of pumilio and pL10 genes was analysed by qPCR and in situ hybridization, showing that the expression of these genes was stronger in germinative cells but not restricted to this cell type. This study provides the first tools to analyse and further characterise undifferentiated cells in a model cestode.

First description of peritoneal and pleural metacestodosis caused by Mesocestoides vogae in a European wild cat (Felis silvestris silvestris).

Tapeworms of the genus Mesocestoides (Cestoda: Cyclophyllidea: Mesocestoididae) are still enigmatic to scientists, due to their high morphological variability, low host specificity, and unknown details of their life cycle. They are found worldwide, with carnivorous mammals as the main definitive hosts, and the disease is potentially zoonotic. After ingestion by a definitive host, the tetrathyridium can occasionally migrate through the intestinal wall and reach the peritoneal cavity or abdominal organs causing peritoneal metacestodosis. Here, we report on a case of metacestodosis of a European wild cat (Felis silvestris silvestris) found dead in Croatia. At necropsy, a large number of white, rice-like structures were found free in the abdominal and thoracic cavities, as well as along the serous surfaces and in the lungs. DNA isolated from the nodules was genotyped and based on a 320-base pair long 12S fragment classified as Mesocestoides vogae. Although post-mortem changes were advanced, severe emaciation due to the severe parasitic infection and gastrointestinal bleeding was diagnosed as the likely cause of death. Intestinal cestodosis was previously reported in wild cats, but according to our knowledge, this is the first description of peritoneal and pleural metacestodosis caused by M. vogae tetrathyridia (metacestodes) in any wild carnivore species.

Clinical forms of peritoneal larval cestodiasis by Mesocestoides spp. in dogs: diagnosis, treatment and long term follow-up.

Canine peritoneal larval cestodiasis (CPLC) is a little-known parasitological infestation of the peritoneal cavity of wild and domestic carnivores with Mesocestoides spp. larvae. While adult Mesocestoides tapeworms reside within the small intestine, the larvae occasionally penetrate the host's intestinal wall, causing a potentially life-threatening peritonitis. Severity of infection as well as the host response influences the prognosis significantly, and early diagnosis and treatment are essential. However, due to the lack of specific symptoms, this condition is underdiagnosed and, furthermore, no clear effective treatment has yet been described. The aim of this study is therefore to report two clinical cases of CPLC in dogs and to illustrate their clinical presentation and follow-up to serve as a reference for clinicians and researchers alike. Both animals were presented with abdominal distention as their main complaint. They underwent clinical examination, abdominal ultrasonography, abdominocentesis, and laparotomy followed by biochemical, cytological, parasitological, and molecular examination of the collected samples. After surgical lavage, the dogs received anthelmintic treatment with either fenbendazole (FBZ) or praziquantel (PZQ). Overall, timely and prolonged administration of high doses of FBZ seems to be the most effective treatment method. Irrespective, to date, no treatment capable of complete eradication of the infection and prevention of recurrence of disease has been found. In conclusion, further investigation into appropriate treatment plans as well as diagnostic development is needed.

The forgotten exotic tapeworms: a review of uncommon zoonotic Cyclophyllidea.

As training in helminthology has declined in the medical microbiology curriculum, many rare species of zoonotic cestodes have fallen into obscurity. Even among specialist practitioners, knowledge of human intestinal cestode infections is often limited to three genera, Taenia, Hymenolepis and Dibothriocephalus. However, five genera of uncommonly encountered zoonotic Cyclophyllidea (Bertiella, Dipylidium, Raillietina, Inermicapsifer and Mesocestoides) may also cause patent intestinal infections in humans worldwide. Due to the limited availability of summarized and taxonomically accurate data, such cases may present a diagnostic dilemma to clinicians and laboratories alike. In this review, historical literature on these cestodes is synthesized and knowledge gaps are highlighted. Clinically relevant taxonomy, nomenclature, life cycles, morphology of human-infecting species are discussed and clarified, along with the clinical presentation, diagnostic features and molecular advances, where available. Due to the limited awareness of these agents and identifying features, it is difficult to assess the true incidence of these 'forgotten' cestodiases as clinical misidentifications are likely to occur. Also, the taxonomic status of many of the human-infecting species of these tapeworms is unclear, hampering accurate species identification. Further studies combining molecular data and morphological observations are necessary to resolve these long-standing taxonomic issues and to elucidate other unknown aspects of transmission and ecology.

Co-administration of silymarin elevates the therapeutic effect of praziquantel through modulation of specific antibody profiles, Th1/Th2/Tregs cytokines and down-regulation of fibrogenesis in mice with Mesocestoides vogae (Cestoda) infection.

Silymarin (SIL) represents a natural mixture of polyphenols showing an array of health benefits. The present study, carried out on a model cestode infection induced by Mesocestoides vogae tetrathyridia in the ICR strain of mice, was aimed at investigating the impact of SIL as adjunct therapy on the activity of praziquantel (PZQ) in relation to parasite burden, immunity and liver fibrosis within 20 days post-therapy. In comparison with PZQ alone, co-administration of SIL and PZQ stimulated production of total IgG antibodies to somatic and excretory-secretory antigens of metacestodes and modified the expression patterns of immunogenic molecules in both antigenic preparations. The combined therapy resulted in the elevation of IFN-γ and a decline of TNF-α and TGF-ß1 in serum as compared to untreated group; however, SIL attenuated significantly the effect of PZQ on IL-4 and stimulated PZQ-suppressed phagocytosis of peritoneal macrophages. In the liver, SIL boosted the effect of PZQ on gene expression of the same cytokines in a similar way as was found in serum, except for down-regulation of PZQ-stimulated TNF-α. Compared to PZQ therapy, the infiltration of mast cells into liver after SIL co-administration was nearly abolished and correlated with suppressed activities of genes for collagen I, collagen III and α-SMA. In conclusion, co-administration of SIL modified the effects of PZQ therapy on antigenic stimulation of the immune system and modulated Th1/Th2/Tregs cytokines. In liver this was accompanied by reduced fibrosis, which correlated with significantly higher reduction of total numbers of tetrathyridia after combined therapy as compared with PZQ treatment.

Comparison of different staining methods for determination of viability on Mesocestoides vogae tetrathyridia.

Mesocestoides vogae is widely employed as a model for studying the biology, differentiation, and experimental chemotherapy of cestodes. Currently, there are few techniques to measure the viability of M. vogae metacestodes during pharmacological experiments. The aim of the present work was to evaluate and compare different staining techniques to determine objectively the viability of M. vogae tetrathyridia. Eosin (0.05% w/v), methylene blue (0.01% w/v), propidium iodide (PI, 2 µg/ml), and fluorescein diacetate (FDA, 0.5 µg/ml) solutions were tested against live, heat-killed (cultivated at 65 °C for 2 h) and thymol-treated tetrathyridia (50 and 250 µg/ml). Parasites were counted under a dissecting microscope or a fluorescence compound microscope, as appropriate. Studies by scanning electron microscope were performed to compare the ultrastructural damage with the viability of parasites. After comparing the performance of different dyes, we chose the eosin staining technique because its simplicity, rapidity, sensitivity, low cost and fidelity.

Urinary incontinence associated with Mesocestoides vogae infection in a dog.

Peritoneal larval cestodiasis caused by Mesocestoides spp. is a rare infection in dogs. A 6-year-old female dog was presented for veterinary care with urinary incontinence which started 1 year earlier. After performing hematology, ultrasound, and computerized tomography, an exploratory laparotomy revealed canine peritoneal larval cestodiasis (CPLC) with the presence of Mesocestoides vogae (syn. Mesocestoides corti) tetrathyridia confirmed by morphological identification and PCR and DNA sequencing. Parasitic cysts were found around the urinary bladder and appeared to inhibit its normal function. An initial treatment with 5 mg/kg praziquantel subcutaneously every 2 weeks for four treatments failed to alleviate the clinical signs, and only treatment with fenbendazole at 100 mg/kg P.O. twice daily for 28 days was associated with the disappearance of ascites and regaining of urinary control. This is the first report of CPLC associated with urinary incontinence in dogs and the first description of this cyclophyllidean cestode in dogs in Israel.

Experimental murine model of neurocysticercosis: first report of cerebellum as a location for Mesocestoides corti tetrathyridia.

Neurocysticercosis is a parasitic disease caused by encysted larvae of Taenia solium in the human central nervous system. Cysts mainly affect the cerebral hemispheres, although they can also be found in ventricles, basal cisterns, and subarachnoid spaces, and rarely in the cerebellum. Given the impossibility of studying the disease in human patients, Cardona et al. (1999) developed a mouse model of neurocysticercosis, using Mesocestoides corti, a closely related cestode. This allows us to study the parasite-host relationship and the mechanisms involved in the disease, in order to improve the therapy. In this murine model of neurocysticercosis, the location of tetrathyridia in parenchyma, ventricles and meninges has already been reported. The aim of this work is to report the cerebellum as a new location for M. corti tetrathyridia in the murine model of neurocysticercosis. A murine model that reproduces the human pathology is essential to evaluate the symptomatology and response to drug treatment in experimentally infected mice.

Gastrointestinal helminths of gray wolves (Canis lupus lupus) from Sweden.

As the Scandinavian wolf population is limited in size, it is only rarely subject to systematic studies on its disease biology, especially gastrointestinal parasites. Therefore, this study aims to describe the prevalence of gastrointestinal helminths of gray wolves hunted on a limited license as a part of a wildlife management program. Helminths of 20 wolves were examined post mortem by macroscopy and coprology. Intestinal worms of five species were recovered from 18 wolves (90%): Uncinaria stenocephala (90%), Taenia spp. (45%), Alaria alata (25%), and Mesocestoides spp. (5%). Of the taeniid specimens typed by multiplex PCR and sequencing of the cox1 gene, 25% belonged to Taenia hydatigena and 25% to Taenia krabbei. The overall species diversity was low compared to findings from wolves of the northern hemisphere. Fecal eggs of Eucoleus boehmi were detected in 12 wolves (60%). Fecal metastrongylid larvae were found in seven individuals (39%), but PCR analyses specific for Angiostrongylus vasorum were negative. The wolves were in good body condition suggesting that the parasite infestation had no negative impact on the general health of the examined wolves. Although some of the recovered parasite species have zoonotic or veterinary impact, it is not likely that the spare wolf population pose substantial threat to human or veterinary health.

Differential Effects of the Flavonolignans Silybin, Silychristin and 2,3-Dehydrosilybin on Mesocestoides vogae Larvae (Cestoda) under Hypoxic and Aerobic In Vitro Conditions.

Mesocestoides vogae larvae represent a suitable model for evaluating the larvicidal potential of various compounds. In this study we investigated the in vitro effects of three natural flavonolignans-silybin (SB), 2,3-dehydrosilybin (DHSB) and silychristin (SCH)-on M. vogae larvae at concentrations of 5 and 50 µM under aerobic and hypoxic conditions for 72 h. With both kinds of treatment, the viability and motility of larvae remained unchanged, metabolic activity, neutral red uptake and concentrations of neutral lipids were reduced, in contrast with a significantly elevated glucose content. Incubation conditions modified the effects of individual FLs depending on their concentration. Under both sets of conditions, SB and SCH suppressed metabolic activity, the concentration of glucose, lipids and partially motility more at 50 µM, but neutral red uptake was elevated. DHSB exerted larvicidal activity and affected motility and neutral lipid concentrations differently depending on the cultivation conditions, whereas it decreased glucose concentration. DHSB at the 50 µM concentration caused irreversible morphological alterations along with damage to the microvillus surface of larvae, which was accompanied by unregulated neutral red uptake. In conclusion, SB and SCH suppressed mitochondrial functions and energy stores, inducing a physiological misbalance, whereas DHSB exhibited a direct larvicidal effect due to damage to the tegument and complete disruption of larval physiology and metabolism.

Spatio-temporal expression of Mesocestoides corti McVAL2 during strobilar development.

VAL proteins belong to a diverse superfamily containing the CAP domain, with members described for various eukaryotic organisms, including parasites. They are implicated in diverse biological activities and, as secreted proteins, may be related in host - parasite interactions. For this reason they have been proposed as vaccine candidates against nematode infections. However, little is known about their function in cestodes. In M. corti, four partial cDNA sequences coding for members of the CAP superfamily were previously isolated. In this work we characterize the expression of McVAL2 in the larvae and segmented worms of M. corti, describing mRNA and protein localization using fluorescent microscopy. We also optimized real time PCR analysis for quantification of mRNA expression through the different stages of strobilar development. We show that McVAL2 is differentially located, depending on the developmental stage, and can be used as a molecular marker for the neuroendocrine system in the larvae. The dynamic and stage-specific expression patterns of McVAL2, combined with the large number of VAL proteins found in the genomes of parasitic flatworms, suggest varied roles for the VAL protein family in the biology of these parasites.

Purification of native M. vogae and H. contortus tubulin by TOG affinity chromatography.

Microtubules are non-covalent cylindrical polymers formed by alpha- and beta-tubulin heterodimer units, crucial for cell division, intracellular transport, motility and differentiation. This makes them very attractive pharmacological targets exploited to develop different drugs such as anthelmintics, antifungals, and antineoplastics. In this work, in order to establish an in vitro target-based screen to integrate to the search for new anthelmintics, we explored the extraction of native assembly-competent tubulin from two helminth parasites: Mesocestoides vogae tetrathyridia (syn. corti, Cestoda: Cyclophyllidea), a useful cestode biological model, and Haemonchus contortus, a sheep gastrointestinal nematode of interest in livestock production. For this purpose, a novel tubulin affinity chromatography procedure was employed, based on the binding capacity of TOG (Tumor Overexpressed Gene) domain from MAPs (microtubule-associated proteins). The TOG domain of the protein Stu2 from Saccharomyces cerevisiae fused to GST (glutathione S- transferase) were produced in E. coli, and the immobilized recombinant proteins allowed for native tubulin extraction from parasites. The binding capacity of TOG1 affinity column (3.6%) was estimated using commercial porcine brain tubulin. A total amount of up to 126 µg of M. vogae tubulin was purified, whereas H. contortus tubulin co-eluted with glutamate dehydrogenase enzyme. The identity of tubulins was confirmed by western blotting and mass spectrometry. The abundance of tubulin estimated in M. vogae was 10% soluble extract, which probably could explain differences observed between tubulin purification results of both helminth parasites.

In vitro culture of Mesocestoides corti metacestodes and isolation of immunomodulatory excretory-secretory products.

Cestode-mediated diseases hold the interesting feature of persisting metacestode larvae dwelling within the host tissues, in the midst of the immune response. Excretory-secretory (ES) products of the metacestode larval stage modulate the host immune response and modify the outcome of the disease. Therefore, isolation and analysis of axenic metacestode ES products are crucial to study their properties. Here, we report the development of a system for long-term in vitro cultivation of the metacestode of the parasitic cestode Mesocestoides corti (syn. Mesocestoides vogae). Although feeder cells and host serum supported the early growth of the parasite, long-term survival was not dependent on host serum or host-derived factors enabling the collection of parasite released products in serum-free medium. Functionally, these axenic ES products recapitulated M. corti tetrathyridia's ability to inhibit LPS-driven IL-12p70 secretion by dendritic cells. Thus, our new axenic culture system will simplify the identification and characterization of M. corti-derived immunomodulatory factors that will indirectly enable the identification and characterization of corresponding factors in the metacestode larvae of medically relevant cestodes such as Echinococcus multilocularis that are not yet amenable to serum-free cultivation.

Morphological characteristics of Mesocestoides canislagopodis (Krabbe 1865) tetrathyridia found in rock ptarmigan (Lagopus muta) in Iceland.

Necropsies of 1010 rock ptarmigans (Lagopus muta) sampled in autumn 2006-2015 in northeast Iceland revealed Mesocestoides canislagopodis tetrathyridia infections in six birds (0.6 %), two juvenile birds (3 month old), and four adult birds (15 months or older). Four birds had tetrathyridia in the body cavity, one bird in the liver, and one bird both in the body cavity and the liver. There were more tetrathyridia in the body cavity of the two juveniles (c. 50 in each) than in three adults (10-40), possibly indicating a host-age-related tetrathyridia mortality. Approximately, half of tetrathyridia in the body cavity were free or loosely attached to the serosa, the other half were encapsulated in a thin, loose connective tissue stroma, frequently attached to the lungs and the liver. Tetrathyridia in the liver parenchyma incited variably intense inflammation. Tetrathyridia from the juvenile hosts were whitish, heart-shaped, and flattened, with unsegmented bodies with a slightly pointed posterior end. In the adult hosts, tetrathyridia were sometimes almost rectangular-shaped, slightly wider compared to those in the juveniles, but more than twice as long as the younger-aged tetrathyridia. Tetrathyridia infections are most likely acquired during the brief insectivorous feeding phase of ptarmigan chicks, and the tetrathyridia persist throughout the lifespan of the birds.

Occurrence of Mesocestoides canislagopodis (Rudolphi, 1810) (Krabbe, 1865) in mammals and birds in Iceland and its molecular discrimination within the Mesocestoides species complex.

The life cycle of Mesocestoides tapeworms (Cestoda: Cyclophyllidea: Mesocestoididae) requires three hosts. The first intermediate host is unknown but believed to be an arthropod. The second intermediate host is a vertebrate. The primary definitive host is a carnivore mammal, or a bird of prey, that eats the tetrathyridium-infected second intermediate host. One representative of the genus, Mesocestoides canislagopodis, has been reported from Iceland. It is common in the arctic fox (Vulpes lagopus) and has also been detected in domestic dogs (Canis familiaris) and cats (Felis domestica). Recently, scolices of a non-maturing Mesocestoides sp. have also been detected in gyrfalcon (Falco rusticolus) intestines, and tetrathyridia in the body cavity of rock ptarmigan (Lagopus muta). We examined the taxonomic relationship of Mesocestoides from arctic fox, gyrfalcon, and rock ptarmigan using molecular methods, both at the generic level (D1 domain LSU ribosomal DNA) and at the specific level (cytochrome c oxidase subunit I (COI) and 12S mitochondrial DNA). All stages belonged to Mesocestoides canislagopodis. Phylogenetic analysis of the combined 12S-COI at the specific level confirmed that M. canislagopodis forms a distinct clade, well separated from three other recognized representatives of the genus, M. litteratus, M. lineatus, and M. corti/vogae. This is the first molecular description of this species. The rock ptarmigan is a new second intermediate host record, and the gyrfalcon a new primary definitive host record. However, the adult stage seemed not to be able to mature in the gyrfalcon, and successful development is probably restricted to mammalian hosts.

Expression of the histone chaperone SET/TAF-Iß during the strobilation process of Mesocestoides corti (Platyhelminthes, Cestoda).

The histone chaperone SET/TAF-Iß is implicated in processes of chromatin remodelling and gene expression regulation. It has been associated with the control of developmental processes, but little is known about its function in helminth parasites. In Mesocestoides corti, a partial cDNA sequence related to SET/TAF-Iß was isolated in a screening for genes differentially expressed in larvae (tetrathyridia) and adult worms. Here, the full-length coding sequence of the M. corti SET/TAF-Iß gene was analysed and the encoded protein (McSET/TAF) was compared with orthologous sequences, showing that McSET/TAF can be regarded as a SET/TAF-Iß family member, with a typical nucleosome-assembly protein (NAP) domain and an acidic tail. The expression patterns of the McSET/TAF gene and protein were investigated during the strobilation process by RT-qPCR, using a set of five reference genes, and by immunoblot and immunofluorescence, using monospecific polyclonal antibodies. A gradual increase in McSET/TAF transcripts and McSET/TAF protein was observed upon development induction by trypsin, demonstrating McSET/TAF differential expression during strobilation. These results provided the first evidence for the involvement of a protein from the NAP family of epigenetic effectors in the regulation of cestode development.

Inhibition of Tapeworm Thioredoxin and Glutathione Pathways by an Oxadiazole N-Oxide Leads to Reduced Mesocestoides vogae Infection Burden in Mice.

Parasitic flatworms cause serious infectious diseases that affect humans and livestock in vast regions of the world, yet there are few effective drugs to treat them. Thioredoxin glutathione reductase (TGR) is an essential enzyme for redox homeostasis in flatworm parasites and a promising pharmacological target. We purified to homogeneity and characterized the TGR from the tapeworm Mesocestoides vogae (syn. M. corti). This purification revealed absence of conventional TR and GR. The glutathione reductase activity of the purified TGR exhibits a hysteretic behavior typical of flatworm TGRs. Consistently, M. vogae genome analysis revealed the presence of a selenocysteine-containing TGR and absence of conventional TR and GR. M. vogae thioredoxin and glutathione reductase activities were inhibited by 3,4-bis(phenylsulfonyl)-1,2,5-oxadiazole N2-oxide (VL16E), an oxadiazole N-oxide previously identified as an inhibitor of fluke and tapeworm TGRs. Finally, we show that mice experimentally infected with M. vogae tetrathyridia and treated with either praziquantel, the reference drug for flatworm infections, or VL16E exhibited a 28% reduction of intraperitoneal larvae numbers compared to vehicle treated mice. Our results show that oxadiazole N-oxide is a promising chemotype in vivo and highlights the convenience of M. vogae as a model for rapid assessment of tapeworm infections in vivo.

A new approach for the characterization of proliferative cells in cestodes.

Cestodes show a remarkable proliferative capability that sustains the constant growth and differentiation of proglottids essential for their lifestyle. It is believed that a separate population of undifferentiated stem cells (the so-called germinative cells) are the only cells capable of proliferation during growth and development. The study of this particular cell subpopulation is hampered by the current lack of methods to isolate it. In this work, we developed a reproducible flow cytometry and cell sorting method to quantify and isolate the proliferating cells in the tetrathyridia larvae of the model cestode Mesocestoides corti, based on the DNA content of the cells. The isolated cells display the typical germinative cell morphology, and can be used for RNA isolation with a yield in the ng to µg range. We expect that this approach may facilitate the characterization of the germinative cells in M. corti and other model tapeworms.