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A rare case of human taeniasis caused by Taenia saginata with species undetermined cysticercosis

Published online by Cambridge University Press:  19 December 2022

Jie Hou
Affiliation:
Department of Laboratory Medicine, West China Hospital, Sichuan University, Chengdu, Sichuan, China
Weilin Chen
Affiliation:
Department of Laboratory Medicine, West China Hospital, Sichuan University, Chengdu, Sichuan, China
Rong Chen
Affiliation:
Department of Clinical Laboratory, The First People's Hospital of Shuangliu District, Chengdu/West China (Airport) Hospital, Sichuan University, Chengdu, Sichuan, China
Chunlei He
Affiliation:
West China School of Medicine, Sichuan University, Chengdu, Sichuan, China
Ying Ma*
Affiliation:
Department of Laboratory Medicine, West China Hospital, Sichuan University, Chengdu, Sichuan, China
Junyan Qu*
Affiliation:
Center of Infectious Disease, West China Hospital, Sichuan University, Chengdu, China
*
Authors for correspondence: Ying Ma, E-mail: [email protected]; Junyan Qu, E-mail: [email protected]
Authors for correspondence: Ying Ma, E-mail: [email protected]; Junyan Qu, E-mail: [email protected]

Abstract

Taeniasis and cysticercosis, which are caused by Taenia saginata, Taenia solium and Taenia asiatica, are zoonotic parasitic infections with a significant disease burden worldwide. There is consensus amongst experts that T. saginata is a common tapeworm that causes taeniasis in humans as opposed to cysticercosis. This case study of a middle-aged Tibetan man conducted in 2021 challenges the prevailing notion that T. saginata exclusively causes taeniasis and not cysticercosis by documenting symptoms and laboratory studies related to both taeniasis and multiple cysticercosis. The patient's medical record with the symptoms of taeniasis and cysticercosis was reviewed, and the tapeworm's proglottids and cyst were identified from the patient by morphological evaluation, DNA amplification and sequencing. The patient frequently experienced severe headaches and vomiting. Both routine blood screenings and testing for antibodies against the most common parasites were normal. After anthelmintic treatment, an adult tapeworm was found in feces, and medical imaging examinations suggested multiple focal nodules in the brain and muscles of the patient. The morphological and molecular diagnosis of the proglottids revealed the Cestoda was T. saginata. Despite the challenges presented by the cyst's morphology, the molecular analysis suggested that it was most likely T. saginata. This case study suggests that T. saginata infection in humans has the potential to cause human cysticercosis. However, such a conclusion needs to be vetted by accurate genome-wide analysis in patients with T. saginata taeniasis associated with cysts. Such studies shall provide new insights into the pathogenicity of T. saginata.

Type
Research Article
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Copyright
Copyright © The Author(s), 2022. Published by Cambridge University Press

Introduction

The zoonotic infection taeniasis is most commonly caused by Taenia saginata, Taenia solium and Taenia asiatica, which are associated with ingesting raw or undercooked, infected beef meat, pork meat or pork viscera, respectively (Ito et al., Reference Ito, Nakao and Wandra2003; Eichenberger et al., Reference Eichenberger, Thomas, Gabriel, Bobic, Devleesschauwer, Robertson, Saratsis, Torgerson, Braae, Dermauw and Dorny2020). The life cycle of pathogenic Taenia species involves humans as definitive hosts, who harbour the adult stage in their small intestines. Cattle (T. saginata) or pigs (T. solium and T. asiatica) can act as intermediate hosts, who hold the larval stages as cysts that dwell themselves under the skin and in the muscles, nervous system or visceral organs such as the liver of the hosts (Qian et al., Reference Qian, Xiao, Li, Abela-Ridder, Carabin, Fahrion, Engels and Zhou2020). Tapeworm eggs or gravid proglottids are passed in feces, and the eggs can survive for days to months in the environment (Jansen et al., Reference Jansen, Dorny, Gabriël, Dermauw, Johansen and Trevisan2021). Humans can also serve as accidental intermediate hosts when exposed to embryonated eggs (this role is normally fulfilled by T. solium) released by themselves (autoinfestation) or by another tapeworm carrier living in close contact with the subject or involved in contaminated food and water, causing infection in various organs of the human body with the larvae also known as cysticerci (human cysticercosis) (Garcia et al., Reference Garcia, Gonzalez, Evans, Gilman and Cysticercosis Working Group in Peru2003; Garcia, Reference Garcia2018; Alroy and Gilman, Reference Alroy, Gilman, Ryan, Hill, Solomon, Aronson and Endy2020). While T. saginata is responsible for a significant number of cases of taeniasis in humans, it is not thought to be the aetiological agent of cysticercosis in humans because they do not appear to be an accidental intermediate host for this parasite (Garcia et al., Reference Garcia, Gonzalez and Gilman2020). It is not known whether T. asiatica causes cysticercosis in humans (Eom et al., Reference Eom, Rim and Jeon2020).

Twelve cases of T. saginata cysticercosis in humans had been described in the literature before 1972, and these diagnoses were based on autopsy and morphological examination by hookless scoleces (De Rivas, Reference De Rivas1937; Tanasescu and Repciuc, Reference Tanasescu and Repciuc1939; Asenjo and Bustamente, Reference Asenjo and Bustamente1950; Niiio, Reference Niiio1950; Bacigalupo, Reference Bacigalupo JADAB1956; Abuladze, Reference Abuladze1964; Goldsmid, Reference Goldsmid1966; Pawlowski and Schultz, Reference Pawlowski and Schultz1972). In addition, T. saginata infection in adults can occur simultaneously with human cysticercosis (Pawlowski and Schultz, Reference Pawlowski and Schultz1972). The infection of humans with the larvae (cysticerci) of T. saginata appears to be an exceptional situation (Pawlowski and Schultz, Reference Pawlowski and Schultz1972). Medical imaging testing, clinical/exposure criteria, serologic testing, histological pathology and molecular identification all help in the diagnosis of human cysticercosis (Del Brutto et al., Reference Del Brutto, Nash, White, Rajshekhar, Wilkins, Singh, Vasquez, Salgado, Gilman and Garcia2017). Due to the lack of molecular methods readily available in most hospital settings, proper species identification in all reported cases of human cysticercosis is impossible.

Numerous molecular procedures have been developed to identify Taenia spp. responsible for human taeniasis, using mitochondrial and ribosomal DNA, repetitive DNA sequences and/or genes encoding relevant antigens as specimens (Flores et al., Reference Flores, Gonzalez, Hurtado, Motta, Dominguez-Hidalgo, Merino, Perteguer and Garate2018). The cytochrome c oxidase I (COX1) gene is a widely used DNA barcoding marker because the rapid rate of evolution allows for the differentiation of not just closely related species, such as the Taenia genus, but also phylogeographic groups within a single species (Hebert et al., Reference Hebert, Cywinska, Ball and deWaard2003; Galimberti et al., Reference Galimberti, Romano, Genchi, Paoloni, Vercillo, Bizzarri, Sassera, Bandi, Genchi, Ragni and Casiraghi2012; Zhang et al., Reference Zhang, Chen, Yang, Liu, Jiang, Gu, Wang and Wang2014). Evidence suggests the HDP2 segment is a ribosomal DNA sequence useful for species-specific molecular diagnosis of human intestinal taeniasis (Gonzalez et al., Reference Gonzalez, Montero, Harrison, Parkhouse and Garate2000; Flores et al., Reference Flores, Gonzalez, Hurtado, Motta, Dominguez-Hidalgo, Merino, Perteguer and Garate2018). In this case study, we identified the case of a middle-aged Tibetan man with taeniasis owing to T. saginata and cysticercosis that had impacted his brain and muscles diagnosed by morphological and molecular methods. We also used COX1 to create a phylogenetic tree of Taenia.

Materials and methods

Clinical examinations

The electronic medical records of the study subject were retrospectively reviewed, and the following demographic and clinical information was collected: age, gender, past medical history, physical examinations, diagnosis, underlying conditions, drug use, iatrogenic procedures, laboratory tests, computed tomography (CT), enhanced magnetic resonance imaging (MRI), ultrasound scan and clinical outcomes.

Morphological evaluation

Two obvious cysts were detected in the muscles under the right anterior thoracic wall and right midaxillary line through palpation and ultrasound scan, respectively. The cyst biopsy of the right midaxillary line and the proglottids from the patient after deworming treatment was collected and observed morphologically. The gravid proglottids and cystic lesions were fixed in a 10% formalin solution, stained with the hydrochloric acid-carmine solutions and observed under a microscope for pathological diagnosis.

Amplification and sequencing

A portion of the cyst isolated from the right midaxillary line and proglottids was stored in 70% ethanol at −20°C until subsequent molecular diagnosis. Total genomic DNA was extracted by using the Trelief™ Animal Genomic DNA Kit (Tsingke Biotechnology, Beijing, China) in accordance with the manufacturer's instructions. Mitochondrial DNA was analysed by polymerase chain reaction (PCR) targeting COX1 using species-specific forward primers Tsag_COX1/F for T. saginata (positions 322–348) and a common reverse primer Tae_COX1/R for Taenia (positions 1148–1129) (Yamasaki et al., Reference Yamasaki, Allan, Sato, Nakao, Sako, Nakaya, Qiu, Mamuti, Craig and Ito2004), cestode forward primers JB3 (positions 1–24) and reverse primers JB4.5 (positions 444–421) (Bowles et al., Reference Bowles, Blair and McManus1992; Bowles and McManus, Reference Bowles and McManus1994; Tappe et al., Reference Tappe, Berkholz, Mahlke, Lobeck, Nagel, Haeupler, Muntau, Racz and Poppert2016). The HDP2 fragment was amplified with the forward primer PTs7S35F1 and the reverse primer PTs7S35R1 (Gonzalez et al., Reference Gonzalez, Montero, Harrison, Parkhouse and Garate2000). Details of the PCR primers and the reaction conditions are provided in Table 1. PCRs were run in a 30-μL reaction mixture composed of 15-μL I-5™ 2× High-Fidelity Master Mix (Tsingke Biotechnology), 1-μL DNA template, 10 pmol of each primer and 12-μL double-distilled water. The thermocycler parameters of the PCR amplification were as follows: 98°C for 3 min; 39 cycles composed of denaturation at 98°C for 10 s, annealing according to the primers and elongation at 72°C for 15 s. After 39 cycles, the reaction was terminated with an elongation step at 72°C for 5 min, followed by a final hold at 4°C. The amplicons were observed using a 1.5% agarose gel under ultraviolet light and then purified, after which Sanger sequencing methods were carried out at Tsingke Biotechnology or Sangon Biotech (Shanghai, China). The sequenced amplicons were compared to the GenBank database entries using NCBI BLAST (Johnson et al., Reference Johnson, Zaretskaya, Raytselis, Merezhuk, McGinnis and Madden2008), wherein the organism was limited to Taenia (taxid: 6202) and the query coverage was 100%.

Table 1. Primer pairs used for PCRs

a The per cent identity was obtained from the NCBI BLAST webpage and the organism was limited to the Taenia (taxid: 6202) genus and the query coverage was 100%.

b Only proglottids were successfully amplified.

c Both proglottids and cysts were successfully amplified.

Phylogenetic analysis

Multiple sequence alignment was constructed with Clustal W, Clustal Omega and SnapGene software. Phylogenetic analysis of the sequences of COX1 identified in this study and the reference sequences of Taenia species available in the GenBank database (Table 2) were constructed using the maximum-likelihood method by MAGA X. The bootstrap values were calculated with 2000 replications, and the nucleotide substitution of the Tamura–Nei model was adapted.

Table 2. Reference COX1 sequences of Taenia species in GenBank

Results

Case presentation

A middle-aged Tibetan man from China was admitted to our hospital with a 7-year history of recurring severe headaches and vomiting. The headache had started with vomiting, nausea, dizziness and chest tightness, especially on the right side, without any apparent predisposing cause. Weekly, he had 2 or 3 episodes of non-projectile vomiting without bile or coffee-like particles, and the headaches lasted for nearly a day each time. There were no other symptoms, such as fever, chills, convulsions, seizures, increased intracranial pressure or other discomforts. He had not been treated for these signs and symptoms before admitting to our hospital in December 2020. After being fully diagnosed in our hospital, his comorbidities included hypertension, fatty liver, liver insufficiency, paranasal sinusitis and intestinal bacterial infection. The immunosuppression was not found. Additionally, he spent a significant amount of time in a pastoral area that is an epidemic province for taeniasis, although no tapeworm infections were diagnosed before his admission to our hospital. He used to ingest raw beef, which might have led to the ingestion of the cysticerci of T. saginata. Moreover, as far as pork consumption is concerned, his family's eating habit, like most Han Chinese, were eating it cooked, ruling out the possibility of related parasites.

The routine laboratory blood tests, including eosinophil and lymphocyte count, were completely normal apart from the elevated inflammation markers (C-reactive protein and neutrophil count). Schistosomes, Clonorchis sinensis, Echinococcus granulosus and Toxoplasma gondii antibodies were negative in his serum, as were Cryptococcus capsular antigen and next-generation sequencing results from cerebrospinal fluid. Multiple intracranial nodules affecting the supratentorial and infratentorial cerebral parenchyma were shown in detail on CT and MRI of the head, indicating possible intracranial parasitic infection (Fig. 1). Ultrasound scan confirmed the presence of 2 palpable and soft masses located in muscles, which were approximately 19 × 8 × 15 mm3 under the right chest wall and 26 × 10 × 19 mm3 under the right midaxillary line (Fig. 2). A subsequent biopsy of the mass (Fig. 3) showed larval-like tissue, peripheral fibrous tissue hyperplasia, lymphocytic infiltration and hyaline degeneration. The patient was probably diagnosed with taeniasis and cysticercosis and treated with oral albendazole (400 mg, twice daily) over 2 weeks. Hydrocortisone 50 mg was provided 2 days after the first albendazole treatment to counteract any potential negative effects on the central nervous system. After only 2 days of this antiparasitic treatment, the adult tapeworm was eliminated through the patient's feces (Fig. 3). When compared with the first MRI (half a month before antiparasitic treatment), the second MRI (half a month after antiparasitic treatment) demonstrated a slightly smaller focus (Fig. 1). The headache and vomiting resolved, and the patient remained symptom free over a 3-month follow-up period.

Fig. 1. MRI shows that the largest cystic nodule was located next to the left ventricular triangle: (A–C) half a month before antiparasitic treatment and (D–F) half a month after antiparasitic treatment.

Fig. 2. Ultrasound scan shows hypoechoic mass containing cysticercus with calcification in the muscles of (A, B) right anterior thoracic wall (19 × 8 × 15 mm3) and (C, D) right midaxillary line (26 × 10 × 19 mm3).

Fig. 3. Tapeworm materials from the patient: (A) subcutaneous mass; (B) cyst; (C) adult tapeworm; (D) unstained proglottids; (E) a stained proglottid and (F) cystic lesion with hydrochloric acid-carmine staining showing presumed deciduous hooks (black arrows).

Morphological observation

The adult tapeworm was milky white, flat and long like a belt, thin and translucent, and the main segments are shown in Fig. 3. The uterine branches in the gravid proglottids were regular, and each side had approximately 15–30 branches (Fig. 3). The slide of the cyst showed no obvious typical features; however, some presumed deciduous hooks were found in the same slide, leading to indistinguishable morphology (Fig. 3).

Molecular identification

The proglottid from the patient was successfully amplified and sequenced using 2 pairs of the primers of the target COX1. The cyst was amplified with JB3 and JB4.5 primers, and the nuclear DNA HDP2 target fragment was also effectively amplified and sequenced. The 827-bp amplified products of proglottid using the T. saginata species-specific forward primer Tsag_COX1/F and Taenia common reverse primer Tae_COX1/R were 99.27–100% identical to T. saginata and 96.25–96.37% equivalent to T. asiatica without other species (Fig. 4). BLAST results indicated that the per cent identity of the 396-bp PCR products of the proglottid and cyst samples, which had trimmed JB3 and JB4.5 primer sequences, was 99.24–100% identical with T. saginata (Fig. 4). However, the 599 bp of the HDP2 target sequence revealed 99.67–99.83% identity with T. saginata, 95.66–99.67% with T. asiatica and 92.99–97.33% with T. solium under the same BLAST conditions (Fig. 4). Additional information regarding sequence alignment is provided in the Supplementary material. Based on the COX1 phylogenetic analysis (Fig. 5), the case is most likely T. saginata, next T. asiatica and finally T. solium.

Fig. 4. Sequence alignment analysis of (A) COX1 based on Tsag_COX1/F and Tae_COX1/R primers, (B) COX1 based on JB3 and JB4.5 primers and (C) HDP2 sequence.

Fig. 5. Phylogenetic tree was constructed based on the COX1 sequences of Taenia spp. using the maximum-likelihood method.

Discussion

Taeniasis coexisting with cysticercosis in humans is uncommon, especially for T. saginata infection. New insights into the pathogenicity and life cycle of T. saginata may be drawn from the case of the Tibetan patient with Taenia intestinal infection and cysticercosis (brain, muscles) reported in this paper. The life cycle of Taenia, involving the pathogenic mechanism of Taenia infection, is significant for ascertaining the disease classification (taeniasis or cysticercosis), epidemiological context, disease control and prevention (Garcia et al., Reference Garcia, Gonzalez and Gilman2020). Current medical evidence confirmed the proven diagnosis of human taeniasis caused by T. saginata and suggested the diagnosis of muscular cysticercosis and neurocysticercosis caused by the larval stage of uncertain Taenia species, which may belong to T. saginata, T. asiatica or hybrids of the 2 sibling species (Okamoto et al., Reference Okamoto, Nakao, Blair, Anantaphruti, Waikagul and Ito2010; Yamane et al., Reference Yamane, Suzuki, Tachi, Li, Chen, Nakao, Nkouawa, Yanagida, Sako, Ito, Sato and Okamoto2012). Studies have also reported that rare Taenia species (Taenia crassiceps, Taenia serialis and Taenia martis) also cause human cysticercosis, confirmed by molecular diagnostic tests (Tappe et al., Reference Tappe, Berkholz, Mahlke, Lobeck, Nagel, Haeupler, Muntau, Racz and Poppert2016; Mueller et al., Reference Mueller, Förch, Zustin, Muntau, Schuldt and Tappe2020). The potential pathogenetic mechanism of the larval stage of these species might be similar to T. solium, wherein embryos hatch in the small intestine and then may invade and spread haematogenously to the brain or muscle and other tissues (Garcia et al., Reference Garcia, Gonzalez and Gilman2020).

Three common Taenia species can be differentiated by examination of their morphological characteristics, such as the scolex, mature and gravid proglottids in the adult stage, and the scolex in the larval stage (Eom et al., Reference Eom, Rim and Jeon2020). However, the differential diagnosis between them is challenging when their morphology is not so typically visible. As far as morphological characteristics were concerned, the gravid proglottids (Fig. 3) from this patient were the same as T. saginata or T. asiatica, not T. solium, while the morphological evaluation of cysticercosis, that showed the presence of detached hooks (Fig. 3), referred to T. asiatica and T. solium rather than T. saginata. However, the traditional morphological taxonomy has some inherent limitations leading to possible false-species identification and can neglect cryptic or pseudocryptic species. That is the reason why integrated taxonomy approaches, wherein molecular studies combine detailed morphological information, are important in helping characterize pairing at the individual level resulting in the perfect characterization of cryptic biodiversity (Hebert et al., Reference Hebert, Cywinska, Ball and deWaard2003; Laakmann et al., Reference Laakmann, Blanco-Bercial and Cornils2020).

The most common method for molecular identification of Taenia tapeworms has been PCR coupled with the sequencing of the amplified PCR product (Ale et al., Reference Ale, Victor, Praet, Gabriël, Speybroeck, Dorny and Devleesschauwer2014). Other assays for the differential diagnosis of Taenia include multiplex PCR (Nunes et al., Reference Nunes, Lima, Manoel, Pereira, Nakano and Garcia2003; Yamasaki et al., Reference Yamasaki, Allan, Sato, Nakao, Sako, Nakaya, Qiu, Mamuti, Craig and Ito2004; Jeon et al., Reference Jeon, Chai, Kong, Waikagul, Insisiengmay, Rim and Eom2009; Ng-Nguyen et al., Reference Ng-Nguyen, Stevenson, Dorny, Gabriel, Vo, Nguyen, Phan, Hii and Traub2017), PCR coupled with restriction fragment length polymorphism analysis (Gonzalez et al., Reference Gonzalez, Montero, Morakote, Puente, Diaz De Tuesta, Serra, Lopez-Velez, McManus, Harrison, Parkhouse and Garate2004), random-amplified polymorphic DNA analysis (Jeon et al., Reference Jeon, Chai, Kong, Waikagul, Insisiengmay, Rim and Eom2009), loop-mediated isothermal amplification (Nkouawa et al., Reference Nkouawa, Sako, Nakao, Nakaya and Ito2009) and matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (Wendel et al., Reference Wendel, Feucherolles, Rehner, Poppert, Utzinger, Becker and Sy2021). Different target sequences of Taenia genomic DNA, including mitochondrial DNA (e.g. COX1, CYTb, valine tRNA, NADH, 12S rDNA gene) (Yamasaki et al., Reference Yamasaki, Nakao, Sako, Nakaya, Sato, Mamuti, Okamoto and Ito2002; Okamoto et al., Reference Okamoto, Nakao, Blair, Anantaphruti, Waikagul and Ito2010; Roelfsema et al., Reference Roelfsema, Nozari, Pinelli and Kortbeek2016), ribosomal DNA (i.e. 28S rRNA, 5.8S rRNA and ITS rRNA gene) (von Nickisch-Rosenegk et al., Reference von Nickisch-Rosenegk, Silva-Gonzalez and Lucius1999; Praet et al., Reference Praet, Verweij, Mwape, Phiri, Muma, Zulu, van Lieshout, Rodriguez-Hidalgo, Benitez-Ortiz, Dorny and Gabriel2013) and nuclear DNA (HDP1, HDP2, cathepsin L-like cysteine peptidase, Ag2 gene) (Gonzalez et al., Reference Gonzalez, Montero, Harrison, Parkhouse and Garate2000; Gonzalez et al., Reference Gonzalez, Montero, Morakote, Puente, Diaz De Tuesta, Serra, Lopez-Velez, McManus, Harrison, Parkhouse and Garate2004; Flores et al., Reference Flores, Gonzalez, Hurtado, Motta, Dominguez-Hidalgo, Merino, Perteguer and Garate2018), have been used as markers in molecular diagnosis. In particular, the mitochondrial COX1 gene that has been used in the study is a universally accepted DNA barcoding marker for assessing the genetic variation and evolutionary biology of Metazoa, including helminth parasites (Hebert et al., Reference Hebert, Cywinska, Ball and deWaard2003; Galimberti et al., Reference Galimberti, Romano, Genchi, Paoloni, Vercillo, Bizzarri, Sassera, Bandi, Genchi, Ragni and Casiraghi2012; Zhang et al., Reference Zhang, Chen, Yang, Liu, Jiang, Gu, Wang and Wang2014; Rostami et al., Reference Rostami, Salavati, Beech, Babaei, Sharbatkhori and Harandi2015; Mioduchowska et al., Reference Mioduchowska, Czyz, Goldyn, Kur and Sell2018; Eberle et al., Reference Eberle, Ahrens, Mayer, Niehuis and Misof2020). Molecular characterization in our study based on COX1 and HDP2 indicated that the cyst seemed to be more inclined to T. saginata or T. asiatica. The analysis of HDP2 sequences in cysticercus samples ruled out the possibility of T. solium, but failed to reliably differentiate T. saginata from T. asiatica or their hybrids (Okamoto et al., Reference Okamoto, Nakao, Blair, Anantaphruti, Waikagul and Ito2010). High homologies were found between T. saginata and T. asiatica on mitochondrial COX1 gene, as shown by the phylogenetic dendrogram (Fig. 5) which is in line with a prior study showing a 4.6% difference between the whole mitochondrial genome sequence of T. saginata and T. asiatica (Jeon et al., Reference Jeon, Kim and Eom2007). However, the BLAST results of the cyst were inconsistent with its morphology, which encompassed dropping hooks in the slide (Fig. 3). These disagreements in morphology and molecular identification suggested the possibility of hybrids of T. saginata and T. asiatica.

For studies involving DNA sequences, whole-genome sequencing has become the standard method for collecting relevant data (Eberle et al., Reference Eberle, Ahrens, Mayer, Niehuis and Misof2020). Unfortunately, due to the scarcity of high-quality cyst samples, we have not been able to sequence the entire cyst genome. However, our study suggested that patients with taeniasis accompanied by cysticercosis may need whole-genome analysis to further ascertain whether T. saginata is also responsible for causing human cysticercosis. Despite the widespread availability of molecular identification methods, often tested on proglottids' samples, the sensitivity of detection in cyst samples may be lowered due to DNA degradation within cysts (Figueiredo et al., Reference Figueiredo, RA, Sato, da Silva, Pereira-Junior, Chigusa, Kawai and Sato2019). Many researchers have reported various mitochondrial/nuclear gene discordances in specimens, indicating potential hybrids between T. asiatica and T. saginata (Nkouawa et al., Reference Nkouawa, Sako, Nakao, Nakaya and Ito2009; Okamoto et al., Reference Okamoto, Nakao, Blair, Anantaphruti, Waikagul and Ito2010; Yamane et al., Reference Yamane, Suzuki, Tachi, Li, Chen, Nakao, Nkouawa, Yanagida, Sako, Ito, Sato and Okamoto2012; Ale et al., Reference Ale, Victor, Praet, Gabriël, Speybroeck, Dorny and Devleesschauwer2014). Therefore, a molecular sequence-based species characterization that relies on information encoded by a single gene of the mitochondrial genome can be deceptive. Hence, mitochondrial, ribosomal or nuclear marker-based multilocus sequence analysis of genetic heterogeneity within the Taenia spp. could be the more effective routine molecular identification in clinical laboratories (Pajuelo et al., Reference Pajuelo, Eguiluz, Dahlstrom, Requena, Guzman, Ramirez, Sheen, Frace, Sammons, Cama, Anzick, Bruno, Mahanty, Wilkins, Nash, Gonzalez, Garcia, Gilman, Porcella, Zimic and Cysticercosis Working Group in Peru2015). Therefore, it would be interesting to develop an easy, simple-to-handle and highly sensitive, multilocus sequences-based molecular diagnostic method for specimen identification of Taenia for validation in clinical settings.

In conclusion, the cause of human cysticercosis by non-solium Taenia species, such as T. saginata or T. asiatica, remains unclear. Correct diagnosis can be aided by utilizing whole-genome analysis and molecular identification approaches that focus on multilocus sequences. In order to better understand the comprehensive epidemiology and total life cycle of Taenia, the limitations of the existing molecular diagnostic applications require more research for the correct diagnosis and prevention of the disease.

Supplementary material

The supplementary material for this article can be found at https://doi.org/10.1017/S003118202200169X

Data availability

The data and materials information used and analyzed in the current study are available from the first author (Jie Hou) or corresponding author (Ying Ma) upon reasonable request.

Acknowledgements

We thank the patient and his family.

Author's contributions

Conceptualization: Jie Hou, Ying Ma and Junyan Qu; data curation: Jie Hou and Weilin Chen; formal analysis: Jie Hou, Weilin Chen and Rong Chen; investigation: Jie Hou, Weilin Chen and Chunlei He; methodology: Jie Hou; project administration: Ying Ma and Jie Hou; resources: Ying Ma and Junyan Qu; software: Jie Hou; supervision: Ying Ma and Junyan Qu; validation: Ying Ma and Junyan Qu; visualization: Jie Hou; writing – original draft: Jie Hou; writing – review and editing: Ying Ma and Junyan Qu.

Financial support

This research received no specific grant from any funding agency, commercial or not-for-profit sectors.

Conflict of interest

The authors declare there are no conflicts of interest.

Ethical standards

The patient's oral informed consent was obtained by phone, and written informed consent was waived with the approval of the West China Hospital, Sichuan University Biomedical Ethics Committee (approval number: 2022-472) because we met the criteria for an informed consent waiver.

References

Abuladze, KI (1964) Principles of Cestology, Vol. IV. Moscow: Nauka, p. 530.Google Scholar
Ale, A, Victor, B, Praet, N, Gabriël, S, Speybroeck, N, Dorny, P and Devleesschauwer, B (2014) Epidemiology and genetic diversity of Taenia asiatica: a systematic review. Parasites & Vectors 7, 45.CrossRefGoogle ScholarPubMed
Alroy, KA and Gilman, RH (2020) Tapeworm infections. In Ryan, Edward T., Hill, David R., Solomon, Tom, Aronson, Naomi E. and Endy, Timothy P. (eds), Hunter's Tropical Medicine and Emerging Infectious Diseases. London: Elsevier, pp. 932940.CrossRefGoogle Scholar
Asenjo, A and Bustamente, E (1950) Die neurochirurgische behandlung der cysticerkose. DMW-Deutsche Medizinische Wochenschrift 75, 11801183.CrossRefGoogle Scholar
Bacigalupo JADAB, A, (1956) La Taenia suginuta puede producir cysticercus bovis en el hombre? Prensa Medica Argentina 43, 10521054.Google Scholar
Bowles, J and McManus, DP (1994) Genetic characterization of the Asian Taenia, a newly described taeniid cestode of humans. American Journal of Tropical Medicine and Hygiene 50, 3344.CrossRefGoogle ScholarPubMed
Bowles, J, Blair, D and McManus, DP (1992) Genetic variants within the genus Echinococcus identified by mitochondrial DNA sequencing. Molecular and Biochemical Parasitology 54, 165173.CrossRefGoogle ScholarPubMed
Del Brutto, OH, Nash, TE, White, AC Jr, Rajshekhar, V, Wilkins, PP, Singh, G, Vasquez, CM, Salgado, P, Gilman, RH and Garcia, HH (2017) Revised diagnostic criteria for neurocysticercosis. Journal of the Neurological Sciences 372, 202210.CrossRefGoogle ScholarPubMed
De Rivas, D (1937) Cysticercus bovis in man. In Papers on helminthology published in communication of the 30 year jubileum of K. J. Skriabin.Google Scholar
Eberle, J, Ahrens, D, Mayer, C, Niehuis, O and Misof, B (2020) A plea for standardized nuclear markers in metazoan DNA taxonomy. Trends in Ecology & Evolution 35, 336345.CrossRefGoogle ScholarPubMed
Eichenberger, RM, Thomas, LF, Gabriel, S, Bobic, B, Devleesschauwer, B, Robertson, LJ, Saratsis, A, Torgerson, PR, Braae, UC, Dermauw, V and Dorny, P (2020) Epidemiology of Taenia saginata taeniosis/cysticercosis: a systematic review of the distribution in east, southeast and south Asia. Parasites & Vectors 13, 234.CrossRefGoogle ScholarPubMed
Eom, KS, Rim, HJ and Jeon, HK (2020) Taenia asiatica: historical overview of taeniasis and cysticercosis with molecular characterization. Advances in Parasitology 108, 133173.CrossRefGoogle ScholarPubMed
Figueiredo, BNS, RA, LI, Sato, M, da Silva, CF, Pereira-Junior, RA, Chigusa, Y, Kawai, S and Sato, MO (2019) Occurrence of bovine cysticercosis in two regions of the state of Tocantins-Brazil and the importance of pathogen identification. Pathogens (Basel, Switzerland) 8(2), 66. doi: 10.3390/pathogens8020066Google ScholarPubMed
Flores, MD, Gonzalez, LM, Hurtado, C, Motta, YM, Dominguez-Hidalgo, C, Merino, FJ, Perteguer, MJ and Garate, T (2018) HDP2: a ribosomal DNA (NTS-ETS) sequence as a target for species-specific molecular diagnosis of intestinal taeniasis in humans. Parasites & Vectors 11, 117.CrossRefGoogle ScholarPubMed
Galimberti, A, Romano, DF, Genchi, M, Paoloni, D, Vercillo, F, Bizzarri, L, Sassera, D, Bandi, C, Genchi, C, Ragni, B and Casiraghi, M (2012) Integrative taxonomy at work: DNA barcoding of taeniids harboured by wild and domestic cats. Molecular Ecology Resources 12, 403413.CrossRefGoogle ScholarPubMed
Garcia, HH (2018) Neurocysticercosis. Neurologic Clinics 36, 851864.CrossRefGoogle ScholarPubMed
Garcia, HH, Gonzalez, AE, Evans, CA, Gilman, RH and Cysticercosis Working Group in Peru, (2003) Taenia solium cysticercosis. Lancet (London, England) 362, 547556.CrossRefGoogle ScholarPubMed
Garcia, HH, Gonzalez, AE and Gilman, RH (2020) Taenia solium cysticercosis and its impact in neurological disease. Clinical Microbiology Reviews 33(3), e0008519. doi: 10.1128/CMR.00085-19CrossRefGoogle ScholarPubMed
Goldsmid, J (1966) Two unusual cases of cysticercosis in man in Rhodesia. Journal of Helminthology 40, 331336.CrossRefGoogle ScholarPubMed
Gonzalez, LM, Montero, E, Harrison, LJ, Parkhouse, RM and Garate, T (2000) Differential diagnosis of Taenia saginata and Taenia solium infection by PCR. Journal of Clinical Microbiology 38, 737744.CrossRefGoogle ScholarPubMed
Gonzalez, LM, Montero, E, Morakote, N, Puente, S, Diaz De Tuesta, JL, Serra, T, Lopez-Velez, R, McManus, DP, Harrison, LJ, Parkhouse, RM and Garate, T (2004) Differential diagnosis of Taenia saginata and Taenia saginata asiatica taeniasis through PCR. Diagnostic Microbiology and Infectious Disease 49, 183188.CrossRefGoogle ScholarPubMed
Hebert, PD, Cywinska, A, Ball, SL and deWaard, JR (2003) Biological identifications through DNA barcodes. Proceedings. Biological Sciences 270, 313321.CrossRefGoogle ScholarPubMed
Ito, A, Nakao, M and Wandra, T (2003) Human taeniasis and cysticercosis in Asia. Lancet (London, England) 362, 19181920.CrossRefGoogle ScholarPubMed
Jansen, F, Dorny, P, Gabriël, S, Dermauw, V, Johansen, MV and Trevisan, C (2021) The survival and dispersal of Taenia eggs in the environment: what are the implications for transmission? A systematic review. Parasites & Vectors 14, 88.CrossRefGoogle ScholarPubMed
Jeon, HK, Kim, KH and Eom, KS (2007) Complete sequence of the mitochondrial genome of Taenia saginata: comparison with T. solium and T. asiatica. Parasitology International 56, 243246.CrossRefGoogle Scholar
Jeon, HK, Chai, JY, Kong, Y, Waikagul, J, Insisiengmay, B, Rim, HJ and Eom, KS (2009) Differential diagnosis of Taenia asiatica using multiplex PCR. Experimental Parasitology 121, 151156.CrossRefGoogle ScholarPubMed
Johnson, M, Zaretskaya, I, Raytselis, Y, Merezhuk, Y, McGinnis, S and Madden, TL (2008) NCBI BLAST: a better web interface. Nucleic Acids Research 36, W5W9.CrossRefGoogle Scholar
Laakmann, S, Blanco-Bercial, L and Cornils, A (2020) The crossover from microscopy to genes in marine diversity: from species to assemblages in marine pelagic copepods. Philosophical Transactions of the Royal Society of London B: Biological Sciences 375, 20190446.CrossRefGoogle ScholarPubMed
Mioduchowska, M, Czyz, MJ, Goldyn, B, Kur, J and Sell, J (2018) Instances of erroneous DNA barcoding of metazoan invertebrates: are universal cox1 gene primers too ‘universal’? PLoS ONE 13, e0199609.CrossRefGoogle ScholarPubMed
Mueller, A, Förch, G, Zustin, J, Muntau, B, Schuldt, G and Tappe, D (2020) Case report: molecular identification of larval Taenia martis infection in the pouch of Douglas. American Journal of Tropical Medicine and Hygiene 103, 23152317.CrossRefGoogle ScholarPubMed
Ng-Nguyen, D, Stevenson, MA, Dorny, P, Gabriel, S, Vo, TV, Nguyen, VT, Phan, TV, Hii, SF and Traub, RJ (2017) Comparison of a new multiplex real-time PCR with the Kato Katz thick smear and copro-antigen ELISA for the detection and differentiation of Taenia spp. in human stools. PLoS Neglected Tropical Diseases 11, e0005743.CrossRefGoogle ScholarPubMed
Niiio, FL (1950) Cisticercosis humana en la Republica Argentina. Estudio de ma nueva observacibn. Prensa Medica Argentina 37, 30403044.Google Scholar
Nkouawa, A, Sako, Y, Nakao, M, Nakaya, K and Ito, A (2009) Loop-mediated isothermal amplification method for differentiation and rapid detection of Taenia species. Journal of Clinical Microbiology 47, 168174.CrossRefGoogle ScholarPubMed
Nunes, CM, Lima, LG, Manoel, CS, Pereira, RN, Nakano, MM and Garcia, JF (2003) Taenia saginata: polymerase chain reaction for taeniasis diagnosis in human fecal samples. Experimental Parasitology 104, 6769.CrossRefGoogle ScholarPubMed
Okamoto, M, Nakao, M, Blair, D, Anantaphruti, MT, Waikagul, J and Ito, A (2010) Evidence of hybridization between Taenia saginata and Taenia asiatica. Parasitology International 59, 7074.CrossRefGoogle ScholarPubMed
Pajuelo, MJ, Eguiluz, M, Dahlstrom, E, Requena, D, Guzman, F, Ramirez, M, Sheen, P, Frace, M, Sammons, S, Cama, V, Anzick, S, Bruno, D, Mahanty, S, Wilkins, P, Nash, T, Gonzalez, A, Garcia, HH, Gilman, RH, Porcella, S, Zimic, M and Cysticercosis Working Group in Peru, (2015) Identification and characterization of microsatellite markers derived from the whole genome analysis of Taenia solium. PLoS Neglected Tropical Diseases 9, e0004316.CrossRefGoogle ScholarPubMed
Pawlowski, Z and Schultz, MG (1972) Taeniasis and cysticercosis (Taenia saginata). Advances in Parasitology 10, 269343.CrossRefGoogle ScholarPubMed
Praet, N, Verweij, JJ, Mwape, KE, Phiri, IK, Muma, JB, Zulu, G, van Lieshout, L, Rodriguez-Hidalgo, R, Benitez-Ortiz, W, Dorny, P and Gabriel, S (2013) Bayesian modelling to estimate the test characteristics of coprology, coproantigen ELISA and a novel real-time PCR for the diagnosis of taeniasis. Tropical Medicine & International Health 18, 608614.CrossRefGoogle Scholar
Qian, MB, Xiao, N, Li, SZ, Abela-Ridder, B, Carabin, H, Fahrion, AS, Engels, D and Zhou, XN (2020) Control of taeniasis and cysticercosis in China. Advances in Parasitology 110, 289317.CrossRefGoogle ScholarPubMed
Roelfsema, JH, Nozari, N, Pinelli, E and Kortbeek, LM (2016) Novel PCRs for differential diagnosis of cestodes. Experimental Parasitology 161, 2026.CrossRefGoogle ScholarPubMed
Rostami, S, Salavati, R, Beech, RN, Babaei, Z, Sharbatkhori, M and Harandi, MF (2015) Genetic variability of Taenia saginata inferred from mitochondrial DNA sequences. Parasitology Research 114, 13651376.CrossRefGoogle ScholarPubMed
Tanasescu, I and Repciuc, E (1939) Ein fall von cysticercus bovis im unterhautgewebe des Menschen. Virchows Archiv für Pathologische Anatomie und Physiologie und für klinische Medizin 304, 555558.Google Scholar
Tappe, D, Berkholz, J, Mahlke, U, Lobeck, H, Nagel, T, Haeupler, A, Muntau, B, Racz, P and Poppert, S (2016) Molecular identification of zoonotic tissue-invasive tapeworm larvae other than Taenia solium in suspected human cysticercosis cases. Journal of Clinical Microbiology 54, 172174.CrossRefGoogle ScholarPubMed
von Nickisch-Rosenegk, M, Silva-Gonzalez, R and Lucius, R (1999) Modification of universal 12S rDNA primers for specific amplification of contaminated Taenia spp. (Cestoda) gDNA enabling phylogenetic studies. Parasitology Research 85, 819825.CrossRefGoogle ScholarPubMed
Wendel, TP, Feucherolles, M, Rehner, J, Poppert, S, Utzinger, J, Becker, SL and Sy, I (2021) Evaluating different storage media for identification of Taenia saginata proglottids using MALDI-TOF mass spectrometry. Microorganisms 9(1), 82. doi: 10.3390/microorganisms9102006CrossRefGoogle ScholarPubMed
Yamane, K, Suzuki, Y, Tachi, E, Li, T, Chen, X, Nakao, M, Nkouawa, A, Yanagida, T, Sako, Y, Ito, A, Sato, H and Okamoto, M (2012) Recent hybridization between Taenia asiatica and Taenia saginata. Parasitology International 61, 351355.CrossRefGoogle ScholarPubMed
Yamasaki, H, Nakao, M, Sako, Y, Nakaya, K, Sato, MO, Mamuti, W, Okamoto, M and Ito, A (2002) DNA differential diagnosis of human taeniid cestodes by base excision sequence scanning thymine-base reader analysis with mitochondrial genes. Journal of Clinical Microbiology 40, 38183821.CrossRefGoogle ScholarPubMed
Yamasaki, H, Allan, JC, Sato, MO, Nakao, M, Sako, Y, Nakaya, K, Qiu, D, Mamuti, W, Craig, PS and Ito, A (2004) DNA differential diagnosis of taeniasis and cysticercosis by multiplex PCR. Journal of Clinical Microbiology 42, 548553.CrossRefGoogle ScholarPubMed
Zhang, G, Chen, J, Yang, Y, Liu, N, Jiang, W, Gu, S, Wang, X and Wang, Z (2014) Utility of DNA barcoding in distinguishing species of the family Taeniidae. Journal of Parasitology 100, 542546.CrossRefGoogle ScholarPubMed
Figure 0

Table 1. Primer pairs used for PCRs

Figure 1

Table 2. Reference COX1 sequences of Taenia species in GenBank

Figure 2

Fig. 1. MRI shows that the largest cystic nodule was located next to the left ventricular triangle: (A–C) half a month before antiparasitic treatment and (D–F) half a month after antiparasitic treatment.

Figure 3

Fig. 2. Ultrasound scan shows hypoechoic mass containing cysticercus with calcification in the muscles of (A, B) right anterior thoracic wall (19 × 8 × 15 mm3) and (C, D) right midaxillary line (26 × 10 × 19 mm3).

Figure 4

Fig. 3. Tapeworm materials from the patient: (A) subcutaneous mass; (B) cyst; (C) adult tapeworm; (D) unstained proglottids; (E) a stained proglottid and (F) cystic lesion with hydrochloric acid-carmine staining showing presumed deciduous hooks (black arrows).

Figure 5

Fig. 4. Sequence alignment analysis of (A) COX1 based on Tsag_COX1/F and Tae_COX1/R primers, (B) COX1 based on JB3 and JB4.5 primers and (C) HDP2 sequence.

Figure 6

Fig. 5. Phylogenetic tree was constructed based on the COX1 sequences of Taenia spp. using the maximum-likelihood method.

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