Using the model cestode Taenia crassiceps for the study of cysticercosis.

Taenia solium is the aetiological agent of taeniasis/cysticercosis, one of the most severe neglected tropical diseases (NTD) according to the World Health Organization (WHO). The life cycle of T. solium alternates between pigs (intermediate host) and humans (definitive host). In addition, humans can act as accidental intermediate hosts if they ingest infective eggs. In this case, the most severe condition of the disease occurs when parasites invade the central nervous system, causing neurocysticercosis (NCC). The complexity of the life cycle of T. solium imposes a barrier to study this pathogen thoroughly. Thus, related species, such as T. crassiceps are commonly used. Due to its capacity to multiply asexually, T. crassiceps can be maintained by serial passage in laboratory mice in standard biosecurity level facilities. In addition, an in vitro system to generate cysticerci in the presence of feeder cells has been recently developed. Despite model species display biological differences with their zoonotic counterparts, they have historically helped to understand the biology of the related pathogenic species and hence, generate improvements in NTD detection and control. In this chapter, we describe the procedures to carry out both in vivo and in vitro systems for T. crassiceps in the laboratory.

Cysticercosis & Heart: A Systematic Review.

Cysticercosis is a parasitic tissue infection caused by larval cysts of the tapeworm Taenia solium. These larval cysts infect brain, muscle, or other tissue, and are a major cause of adult-onset seizures in most low-income countries with tropical climate. Prevalence it's around 50 million people. Although cardiovascular system is not the most affected, this disease can also be associated with multiple and randomly distributed cysts in the subpericardium, subendocardium and myocardium in up to 25% of infected patients. Most cardiac cysticercosis' cases are asymptomatic, but it can manifest with ventricular arrhythmias and conduction disorders. Area Covered: The "Neglected Tropical Diseases and other Infectious Diseases affecting the Heart" (NET-Heart project) is an initiative by the Emerging Leaders group of the Interamerican Society of Cardiology to systematically review all these endemic conditions affecting the heart. A systematic review was conducted following preferred reporting items for systematic review and meta-analysis guidelines and including articles published in MEDLINE, ScienceDirect, PubMed and LILACS databases. A total of 41 papers were included in this review. Expert Opinion: In the areas of greatest prevalence, unhealthiness and poverty favor the development of this disease, which highlights the need to establish global health policies that reduce morbidity and mortality, economic losses of the affected population, and health costs related to hospitalizations for cardiovascular involvement. Authors provide an algorithm to evaluate the possibility of Cysticercosis' cardiovascular complications.

The many facets of disseminated parenchymal brain cysticercosis: A differential diagnosis with important therapeutic implications.

Neurocysticercosis (NCC), the infection of the nervous system by the cystic larvae of Taenia solium, is a highly pleomorphic disease because of differences in the number and anatomical location of lesions, the viability of parasites, and the severity of the host immune response. Most patients with parenchymal brain NCC present with few lesions and a relatively benign clinical course, but massive forms of parenchymal NCC can carry a poor prognosis if not well recognized and inappropriately managed. We present the main presentations of massive parenchymal NCC and their differential characteristics.

Validation of a spatial agent-based model for Taenia solium transmission ("CystiAgent") against a large prospective trial of control strategies in northern Peru.

BACKGROUND: The pork tapeworm (Taenia solium) is a parasitic helminth that imposes a major health and economic burden on poor rural populations around the world. As recognized by the World Health Organization, a key barrier for achieving control of T. solium is the lack of an accurate and validated simulation model with which to study transmission and evaluate available control and elimination strategies. CystiAgent is a spatially-explicit agent based model for T. solium that is unique among T. solium models in its ability to represent key spatial and environmental features of transmission and simulate spatially targeted interventions, such as ring strategy. METHODS/PRINCIPAL FINDINGS: We validated CystiAgent against results from the Ring Strategy Trial (RST)-a large cluster-randomized trial conducted in northern Peru that evaluated six unique interventions for T. solium control in 23 villages. For the validation, each intervention strategy was replicated in CystiAgent, and the simulated prevalences of human taeniasis, porcine cysticercosis, and porcine seroincidence were compared against prevalence estimates from the trial. Results showed that CystiAgent produced declines in transmission in response to each of the six intervention strategies, but overestimated the effect of interventions in the majority of villages; simulated prevalences for human taenasis and porcine cysticercosis at the end of the trial were a median of 0.53 and 5.0 percentages points less than prevalence observed at the end of the trial, respectively. CONCLUSIONS/SIGNIFICANCE: The validation of CystiAgent represented an important step towards developing an accurate and reliable T. solium transmission model that can be deployed to fill critical gaps in our understanding of T. solium transmission and control. To improve model accuracy, future versions would benefit from improved data on pig immunity and resistance, field effectiveness of anti-helminthic treatment, and factors driving spatial clustering of T. solium infections including dispersion and contact with T. solium eggs in the environment.

Taenia solium taeniasis/cysticercosis: From parasite biology and immunology to diagnosis and control.

Infection with the pork tapeworm (Taenia solium) is responsible for a substantial global burden of disease, not only restricted to its impact on human health, but also resulting in a considerable economic burden to smallholder pig farmers due to pig cysticercosis infection. The life-cycle, parasitology and immunology of T. solium are complex, involving pigs (the intermediate host, harbouring the larval metacestode stage), humans (the definitive host, harbouring the adult tapeworm, in addition to acting as accidental intermediate hosts) and the environment (the source of infection with eggs/proglottids). We review the parasitology, immunology, and epidemiology of the infection associated with each of the T. solium life-cycle stages, including the pre-adult/adult tapeworm responsible for human taeniasis; post-oncosphere and cysticercus associated with porcine and human cysticercosis, and the biological characteristics of eggs in the environment. We discuss the burden associated, in endemic settings, with neurocysticercosis (NCC) in humans, and the broader cross-sectoral economic impact associated both with NCC and porcine cysticercosis, the latter impacting food-value chains. Existing tools for diagnostics and control interventions that target different stages of the T. solium transmission cycle are reviewed and their limitations discussed. Currently, no national T. solium control programmes have been established in endemic areas, with further work required to identify optimal strategies according to epidemiological setting. There is increasing evidence suggesting that cross-sectoral interventions which target the parasite in both the human and pig host provide the most effective approaches for achieving control and ultimately elimination. We discuss future avenues for research on T. solium to support the attainment of the goals proposed in the revised World Health Organisation neglected tropical diseases roadmap for 2021-2030 adopted at the 73rd World Health Assembly in November 2020.

Inflammation in neurocysticercosis: clinical relevance and impact on treatment decisions.

INTRODUCTION: Neurocysticercosis is caused by the localization of Taenia solium larvae in the central nervous system. The disease remains endemic in most countries of Latin America, Asia and Africa. While major improvements have been made in its diagnosis and treatment, uncertainties persist regarding the clinical implications and treatment of the inflammatory reaction associated with the disease. AREAS COVERED: In this review, based on PubMed searches, the authors describe the characteristics of the immune-inflammatory response in patients with neurocysticercosis, its clinical implications and the treatment currently administered. The dual role of inflammation (participating in both, the death of the parasite, and the precipitation of serious complications) is discussed. New therapeutic strategies of potential interest are presented. EXPERT OPINION: Inflammatory reaction is the main pathogenic mechanism associated to neurocysticercosis. Its management is mainly based on corticosteroids administration. This strategy had improved prognostic of patients as it allows for the control of most of the inflammatory complications. On the other side, it might be involved in the persistence of parasites in some patients, despite cysticidal treatment, due to its immunosuppressive properties. New strategies are needed to improve therapeutical management, particularly in the severest presentations.

Can We Recommend Practical Interventions to Prevent Neurocysticercosis?

The cystiSim model was used to compare strategies for the control of Taenia solium. A three-monthly intervention in pigs for 3 years was substantially more effective than biannual treatment for taeniasis in the human population for 5 years. The intervention period could be shortened further by combining pig and human interventions.

Transmission of porcine cysticercosis in the Portuguesa state of Venezuela.

The aim of this study was to assess transmission of Taenia solium cysticercosis in Palmarito Arriba, a small village in the rural area of the Portuguesa state of Venezuela, through (1) an evaluation of T. solium transmission risk factors present in the community and (2) serological detection of the secreted metacestode HP10 antigen (HP10 Ag) and of anti-metacestode antibodies in sera from rural pigs. Risk factors associated with transmission of cysticercosis were the following: 100% (23/23) of the households lacked piped water, 87.0% (20/23) of households lacked latrines, 88.0% (100/114) of inhabitants routinely defecated in the open/air, 19.05% (12/63) of the interviewed population had observed proglottids in their stools. More significantly, 9/13 householders breeding pigs reported seeing proglottids in their stools. Of the 25 pigs available for bleeding and serological testing, 64% (16/25) were free roaming and 36% (9/25) were "backyard" animals; 28% (7/25) were seropositive for both the HP10 Ag and antibody, 20.0% (5/25) were seropositive for HP10 Ag alone, and 36.0% (9/25) were seropositive for antibody alone. Given this clear evidence of endemic porcine cysticercosis, further studies are needed to assess and control the level of porcine and human taeniasis and cysticercosis in this and neighboring communities.

Mathematical model of the life cycle of taenia-cysticercosis: transmission dynamics and chemotherapy (Part 1).

BACKGROUND: Taenia solium is the aetiological agent of human taeniasis, pig cysticercosis and human neurocysticercosis, which are serious public health problems, especially in developing countries. METHODS: A mathematical model of the transmission dynamics of taeniasis-cysticercosis is formulated. The model consists of a coupled system of differential equations, which are density-dependent equations for describing the flow of the parasite through the life cycle. The model is hybrid since it comprises deterministic equations with stochastic elements which describe changes in the mean parasite burden and incorporates the overall pattern of the parasites' distribution. RESULTS: Sensitivity and bifurcation analyses were carried out to determine the range of values of the model. The model can reproduce the observed epidemiological patterns of human taeniasis, pig and human cysticercosis. For example, for a wide range of parameter values, the mean intensity of adult worms tends to rapidly stabilize in one parasite per individual host. From this model, we also derived a Susceptible-Infected model to describe the prevalence of infection in humans and pigs. Chemotherapeutic interventions against pig cysticercosis or human taeniasis may reduce rapidly and effectively the mean intensity of human taeniasis, pig cysticercosis and human cysticercosis. This effect can be achieved even if the protective efficacy of the drug is of the order of 90% and the coverage rate is 90%. This means that health in humans infected either with adult worms or cysticerci may be achieved by the application of anthelmintic drugs against pig cysticercosis. However, treatment against human cysticercosis alone, does not influence neither human teniasis nor pig cysticercosis. This is because human cysticercosis infection does not influence the value of the basic reproductive number (Ro). CONCLUSIONS: Even coverage of 100% in the administration of anthelmintics did not eliminate the infection. Then elimination of the infection in all hosts does not seem a feasible goal to achieve by administering only chemotherapeutic interventions. Throughout the manuscript a discussion of our model in the context of other models of taeniasis-cysticercosis is presented.

Spontaneously Arrested Transmission of Cysticercosis in a Highly Endemic Village with a Very Low Migration Rate.

Taenia solium cysticercosis is difficult to eliminate without interventions or societal development. Atahualpa is a rural Ecuadorian village with documented low migration rate, where domestic pig raising is common and human cysticercosis is endemic. To assess neurocysticercosis (NCC) prevalence, 1,273 villagers aged ≥ 20 years underwent neuroimaging studies, which showed calcified lesions in 121 (9.5%) individuals, but no active disease. Likewise, positive reactions, apparently nonspecific, were found in only 3/200 subjects by the use of a monoclonal antibody-based enzyme-linked immunosorbent assay to detect T. solium antigens in urine. Only 2/418 pigs reacted to three antibody bands on serum western blot and none to more than three bands. This is the first time that spontaneously arrested T. solium transmission is documented in a known endemic village. Understanding why active transmission stopped could provide insights on potential targets for control interventions. Atahualpa could provide an optimal scenario for longitudinal studies on the consequences of calcified NCC.

Mapping occurrence of Taenia solium taeniosis/cysticercosis and areas at risk of porcine cysticercosis in Central America and the Caribbean basin.

BACKGROUND: This study aimed to map the occurrence of Taenia solium taeniosis/cysticercosis at national level within Central America and the Caribbean basin, and to map the distribution of porcine cysticercosis at first-level administrative subdivision level (department level) and the porcine population at risk. This zoonotic parasite is believed to be widely endemic across most of Latin America. However, there is little information readily available for Central America and the Caribbean basin. Taenia solium has been ranked the most important foodborne parasitic hazard globally and within endemic areas is a common cause of preventable epilepsy. METHODS: We conducted a structured literature search in PubMed, supplemented and crossed-referenced with relevant academic databases, grey literature, and active searches in identified literature, to identify all records of T. solium presence in Central America and the Caribbean basin between 1986 and April 2017. To retrieve grey literature, government entities, researchers and relevant institutions across the region were contacted in an attempt to cover all countries and territories. Identified records containing data on porcine cysticercosis were geo-referenced to identify department level distribution and compared to modelled distributions of pigs reared under extensive production systems. RESULTS: We identified 51 records of T. solium at the national level, covering 13 countries and an additional three countries were included based on World Organisation for Animal Health (OIE) reports, giving a total of 16 countries out of 41 with evidence of the parasite's presence. Screening records for porcine cysticercosis data at the departmental level confirmed porcine cysticercosis presence in 11 departments across six countries (Colombia, Guatemala, Honduras, Mexico, Nicaragua and Venezuela). CONCLUSIONS: When comparing these results to areas where pigs were kept in extensive production systems and areas where no information on porcine cysticercosis exists, it is apparent that porcine cysticercosis is likely to be underreported, and that a substantial part of the regional pig population could be at risk of contracting porcine cysticercosis. More detailed information on the distribution of T. solium and accurate burden estimations are urgently needed to grasp the true extent of this zoonotic parasite and the public health and agricultural problems it potentially poses.

Role of porcine serum haptoglobin in the host-parasite relationship of Taenia solium cysticercosis.

Human and porcine cysticercosis is a parasitic disease caused by the larval stage (cysts) of the tapeworm Taenia solium. Cysts may live in several host tissues such as skeletal muscle or brain. We have previously described the presence of host haptoglobin (Hp) and hemoglobin (Hb) in different protein extracts of the T. solium cysts. Here, we report the binding of host Hp and Hb to a number of cyst proteins, evaluated through measuring electrophoretic and light absorbance changes. In the sera obtained from 18 cysticercotic pigs, Hp-Hb complexes were abundant, whereas free Hp was undetectable. In contrast, in the sera from non 18 cysticercotic pigs, Hp-Hb and free Hp were found. In the soluble protein fraction of cysts tissue, free Hp was detected showing a considerable Hb-binding ability, whereas in the vesicular fluid, Hp is mainly bound to Hb. Interestingly, assays carried out with the insoluble fraction of T. solium cysts tissue, showed binding of Hp and Hp-Hb in a saturable way, suggesting the existence of specific interactions. Our results suggested that the parasite can take advantage of the uptaken host Hp and Hb, either free or in complexes, as a source of iron or as a way to modulate the inflammatory response surrounding the T. solium cysts.

The endocrine-immune network during taeniosis by Taenia solium: The role of the pituitary gland.

It is well known that sex hormones play an important role during Taenia solium infection; however, to our knowledge no studies exist concerning the immune response following complete or lobe-specific removal of the pituitary gland during T. solium infection. Thus, the aim of this work was to analyze in hamsters, the effects of lack of pituitary hormones on the duodenal immune response, and their impact on T. solium establishment and development. Thus, in order to achieve this goal, we perform anterior pituitary lobectomy (AL, n = 9), neurointermediate pituitary lobectomy (NIL, n = 9) and total hypophysectomy (HYPOX, n = 8), and related to the gut establishment and growth of T. solium, hematoxylin-eosin staining of duodenal tissue and immunofluorescence of duodenal cytokine expression and compared these results to the control intact (n = 8) and control infected group (n = 8). Our results indicate that 15 days post-infection, HYPOX reduces the number and size of intestinally recovered T. solium adults. Using semiquantitative immunofluorescent laser confocal microscopy, we observed that the mean intensity of duodenal IFN-γ and IL-12 Th1 cytokines was mildly expressed in the infected controls, in contrast with the high level of expression of these cytokines in the NIL infected hamsters. Likewise, the duodenum of HYPOX animals showed an increase in the expression of Th2 cytokines IL-5 and IL-6, when compared to control hamsters. Histological analysis of duodenal mucosa from HYPOX hamsters revealed an exacerbated inflammatory infiltrate located along the lamina propria and related to the presence of the parasite. We conclude that lobe-specific pituitary hormones affect differentially the T. solium development and the gut immune response.

Development of specific scFv antibodies to detect neurocysticercosis antigens and potential applications in immunodiagnosis.

We have shown previously that detection of circulating antibodies against mimotopes selected by phage display were useful in neurocysticercosis diagnosis. However, circulating antigens may also be useful in patients' clinical follow-up. Therefore, we aimed to select novel combinatorial antibodies, single-chain variable fragment (scFv), which can be used for specific antigens with pre-defined affinity and specificity without prior immunization. A phage scFv antibody library was selected against Taenia solium mimotopes displayed on phages coupled in beads and total saline extract of T. solium metacestodes (S) immobilized on microtiter plate wells. After two rounds of selection, 96 phage clones were evolved and validated against each target by enzyme linked immunosorbent assay (ELISA), and dot-blot, and three specific antibodies (B6, G10 and A4) were further characterized by sequencing and indirect immunofluorescence (IFI) assays. IFI revealed tegument staining for the B6, while the others showed a non-uniform staining in the whole parasite. The selected scFvs were used to capture their antigen targets that were elucidated through mass spectrometry, and used for antibody detection in NC patients' sera by ELISA, which achieved sensitivities greater than 97% and specificities above 95%. We have successfully developed scFv antibodies against important mimotopes used in NC diagnosis, and can be further explored to detect circulating antigens for clinical follow-up of patients with NC. Our strategy also highlighted the possibility of using this combinatorial approach to select, capture and characterize specific antigens to better understand this intriguing parasite infection and disease evolution.

The highly antigenic 53/25 kDa Taenia solium protein fraction with cathepsin-L like activity is present in the oncosphere/cysticercus and induces non-protective IgG antibodies in pigs.

Cathepsin L-like proteases are secreted by several parasites including Taenia solium. The mechanism used by T. solium oncospheres to degrade and penetrate the intestine and infect the host is incompletely understood. It is assumed that intestinal degradation is driven by the proteolytic activity of enzymes secreted by the oncosphere. Blocking the proteolytic activity by an antibody response would prevent the oncosphere penetration and further infection. Serine and cysteine proteases including chymotrypsin, trypsin, elastase, and cathepsin L, are secreted by T. solium and Taenia saginata oncospheres when cultured in vitro, being potential vaccine candidates. However, the purification of a sufficient quantity of proteases secreted by oncospheres to conduct a vaccine trial is costly and lengthy. A 53/25 kDa cathepsin L-like fraction partially purified from T. solium cyst fluid was described previously as an important antigen for immunodiagnostics. In this study we found that this antigen is present in the T. solium oncosphere and is also secreted by the cysticercus. This protein fraction was tested for its ability to protect pigs against an oral challenge with T. solium oncospheres in a vaccine trial. IgG antibodies against the 53/25 kDa cathepsin L-like protein fraction were elicited in the vaccinated animals but did not confer protection.

Medical management of neurocysticercosis.

INTRODUCTION: Neurocysticercosis (NCC) is considered to be the most common cause of acquired epilepsy worldwide. Formerly restricted to palliative measures, therapy for NCC has advanced with the advent of two drugs that are considered to be effective: praziquantel (PZQ) and albendazole (ALB). AREAS COVERED: All available articles regarding research related to the treatment of NCC were searched. Relevant articles were then reviewed and used as sources of information for this review. EXPERT OPINION: Anticysticercal therapy has been marked by intense controversy. Recent descriptions of spontaneous resolution of parenchymal cysticercosis with benign evolution, risks of complications and reports of no long-term benefits have reinforced the debate over the usefulness and safety of anticysticercal therapy. High interindividual variability and complex pharmacological interactions will require the close monitoring of plasma concentrations of ALB and PZQ metabolites in future trials. Given the relative scarcity of clinical trials, more comparative interventional studies - especially randomized controlled trials in long-term clinical evolution - are required to clarify the controversy over the validity of parasitic therapy in patients with NCC.

Clinical manifestations, diagnosis, and treatment of neurocysticercosis.

Neurocysticercosis (NCC) is the most frequent parasitic disease of the human brain. Modern imaging studies, CT and MRI, have defined the diagnosis and characterization of the disease. Through these studies the therapeutic approach for each case may be individualized with the aid of antihelmintics, steroids, symptomatic medicines, or surgery. The use of one or various therapeutic measures largely depends on the peculiar combination of number, location, and biological stage of lesions as well as the degree of inflammatory response to the parasites. Although there is not a typical clinical picture of NCC, epilepsy is the most frequent manifestation of parenchymal NCC, whereas hydrocephalus is the most frequent manifestation of meningeal NCC. Eradication of cysticercosis is an attainable goal by public education and sanitary improvement in endemic areas.

Hydrocephalus in neurocysticercosis.

INTRODUCTION: Cysticercosis (CC) is the most important of the parasitic diseases of the central nervous system due to its high incidence in the world. CC is the infection with the larval cysts of Taenia solium. It is the most common helminthic infection of the nervous system and is endemic in most underdeveloped countries as well as in industrialized nations. It is estimated that approximately 50,000 people die every year from neurocysticercosis (NCC) worldwide. DISCUSSION: Humans with CC are incidental intermediate hosts, which replace the pig in the life cycle of the T. solium. Children are more frequently affected by parenchyma infestation of cysticercus, of which the main clinical manifestation is epilepsy. Hydrocephalus is more common in adults and is caused by cerebrospinal fluid blockage by ventricular cysts and inflammatory reactions (ependimitis/arachnoiditis). Treatment should be individualized based on clinical presentation, degree of infestation, location and viability of cysticercus, and host response. Hydrocephalus can be controlled only by removal of obstructive intraventricular cysts or associated with either ventriculoperitoneal shunt or endoscopic third ventriculostomy. The degree of infestation and complications related to the shunt represents the most important prognostic factors in the outcome of NCC.

Chinchilla laniger can be used as an experimental model for Taenia solium taeniasis.

Chinchilla laniger has been reported as an experimental definitive host for Taenia solium; however no information about its suitability and yield of gravid tapeworm proglottids containing viable and infective eggs has been published. In total 55 outbred female chinchillas were infected with 4 cysticerci each; hosts were immunodeppressed with 6 or 8 mg of methyl-prednisolone acetate every 14 days starting the day of infection and their discomfort was followed. Kinetics of coproantigen ELISA or expelled proglottids was used to define the infection status. Efficiency of tapeworm establishment was 21% and of parasite gravidity was 8%; chinchillas showed some degree of suffering along the infection. Viability of eggs obtained from gravid proglottids was tested comparing methods previously published, our results showed 62% viability with propidium iodide, 54% with trypan blue, 34% with neutral red, 30% by oncosphere activation and 7% with bromide 3-(4,5-dimetil-tiazol-2-il)-2,5-difenil-tetrazolio (MTT) reduction; no statistical differences were obtained between most techniques, except activation. Four piglets were infected with 50,000 eggs each, necropsy was performed 3 months later and, after counting the number of cysticerci recovered, the percentage of infection was similar to data obtained with T. solium eggs recovered from humans. Our results demonstrate that the experimental model of T. solium taeniasis in C. laniger is a good alternative for providing eggs and adult tapeworms to be used in different types of experiments; optimization of the model probably depends on the use of inbred hosts and on the reduction of infected animals' suffering.