Global epidemiology and molecular biology of Taenia multiceps: a comparative meta-analysis and in silico analysis study.

In the present study, all published data on the epidemiology and molecular characters of Taenia multiceps were systematically collected from relevant databases (e.g. PubMed, Scopus, National Center for Biotechnology Information), and combined in various statistical and genetic analyses as a contribution to a better understanding of the epidemiology of this ubiquitous taeniid worldwide. While 5.8% of the key hosts (dogs) from various countries had T. multiceps, grey wolves displayed the highest prevalence (21.6%) among the definitive hosts. Small ruminants are the main intermediate hosts and carry the coenuri in various locations, but most commonly in the central nervous system (CNS). Cerebral coenuri were confirmed in 53% of sheep exhibiting neurological symptoms, and infected animals often had only a single coenurus in the brain. Sheep had a higher prevalence (8.8%) of CNS coenuri than goats (5.8%); however, extra-CNS coenuri were detected more frequently in goats than in sheep. In either case, the difference between sheep and goats was statistically insignificant. Analysis of 233 partial cytochrome oxidase subunit I nucleotide sections for T. multiceps revealed high haplotype and low nucleotide diversities. Fifty-one haplotypes were detected circulating in 6 geographic populations. China, Iran and Turkey had 2 major haplotypes, whereas Italy and Egypt shared 3. Haplotypes from Greece circulate worldwide, and displayed similar gene flow values when compared with the other populations. There were no distinct patterns for haplotype distribution in relation to the infected hosts or coenuri locations. The existence of genetic variants in T. multiceps was highlighted, but needs further studies.

Taenia multiceps coenurosis: a review.

Taenia multiceps is a taeniid cestode that inhabits the small intestines of both wild and domestic carnivores. The larval stage, Coenurus cerebralis, is typically found in the central nervous system (CNS) of a wide range of livestock and, to a lesser extent, in the extra-cerebral tissues of sheep and goats. This review covers all aspects of the life cycle of T. multiceps and its epidemiology, molecular characterization, pathogenesis, diagnosis, therapy, control and zoonotic potential. Coenurosis caused by the larval stage of T. multiceps has a worldwide distribution and is often fatal in intermediate hosts, which can result in substantial economic losses in livestock farming. Molecular characterization using the mitochondrial genes cytochrome c oxidase subunit 1 and nicotinamide adenine dinucleotide dehydrogenase subunit 1 of different T. multiceps populations has revealed significant genetic variation and the presence of three major haplotypes. The disease mostly affects young sheep and is referred to as either acute or chronic coenurosis. Acute coenurosis occurs as a result of oncospheres migrating through the CNS, while chronic coenurosis occurs as a consequence of the coenurus maturing, which causes displacement and pressure atrophy of brain tissue. Non-cerebral coenurosis has been most commonly reported in goats. The best diagnostic method for cerebral coenurosis involves the interpretation of clinical signs with accurate localization of the cyst using diagnostic imaging techniques. A vaccine based on recombinant oncosphere antigens has proved to be an effective tool against T. multiceps infection in sheep. Additionally, use of anthelmintics during the parasite's migration stages reduces the development of cysts in the sheep brain. Surgery is considered the most effective method for the treatment of cerebral coenurosis in small ruminants, but is often not carried out because of the limited finances of many sheep and goat breeders. However, coenurosis can also be controlled effectively through preventative measures, such as anthelmintic treatment of dogs and the proper disposal of intermediate host carcasses. The parasite is also zoonotic, and cases of coenurosis have been reported in humans with coenuri located in the brain, spinal cord and eyes.

Distribution and prevalence of Taenia hydatigena, Taenia multiceps, and Mesocestoides spp. in Mongolian sheepdogs.

Gastrointestinal parasite infections in livestock and companion animals in Mongolia have not been investigated sufficiently. This study aimed to determine the prevalence of cyclophyllid cestodes (Taeniidae and Mesocestoididae) in Mongolian sheepdogs using copro-DNA analysis. Sheepdog fecal samples (n = 1242) were collected from five ecological zones (mountain taiga, forest-steppe, steppe, desert-steppe, and desert) and four geographical regions (Western, Khangai, Central, and Eastern) within 20 of the country's 21 provinces. Among the 1242 samples, 201 (16.2%) tested positive for mitochondrial cytochrome c oxidase subunit 1 (cox1) and ribosomal 12S rRNA encoding genes of cyclophyllid cestodes. Prevalence in the mountain taiga, forest-steppe, steppe, desert-steppe, and desert zones was 29.2%, 15.4%, 15.1%, 20.1%, and 9.1%, respectively. Prevalence in the Western, Khangai, Central, and Eastern regions was 19.9%, 18.6%, 12.1%, and 12.8%, respectively. Taenia hydatigena, Taenia multiceps, and two Mesocestoides species (Mesocestoides sp.1 and Mesocestoides sp.2) were identified. T. hydatigena was found in the samples from all 20 provinces (all five zones and four regions), while T. multiceps was detected in the samples from 19 provinces (all five zones and four regions). Mesocestoides sp.1 infection was detected in the samples from all zones (except desert) and regions; it was detected in 14 provinces. Mesocestoides sp.2 infection was detected in the samples from all zones (except mountain taiga) and regions and found in seven provinces. Cyclophyllidea infection in sheepdogs is highly prevalent across Mongolia, representing a zoonotic risk. Implementation of a surveillance program for sheepdogs and their owners, and the wild animals in all ecological zones should be considered. In addition, control measures, including public awareness campaigns, especially for sheepdog owners, and periodic deworming of sheepdogs are warranted.

Proteomic characterization of Echinococcus granulosus sensu stricto, Taenia hydatigena and Taenia multiceps metacestode cyst fluids.

Cystic echinococcosis (CE) diagnosis by means of serological assays is hampered by the presence of parasites closely related to Echinococcus granulosus sensu lato (s.l.), responsible of the zoonotic disease and with which share cross-reacting antigens. Thus, improvements on the characterization of Echinococcus specific antigens expressed in the larval stage are required, in order to provide useful information for the development of immunological assays for the serodiagnosis of CE in sheep. Here, the proteome of the hydatid cyst fluids (HFs) of Echinococcus granulosus (hydatid fluid, EgHF) and other ovine parasites cyst fluids (CFs), Taenia hydatigena (ThCF) and Taenia multiceps (TmCF) were analyzed by a shotgun proteomic approach. Parasite and host protein profiles in the three types of cyst fluids were characterized and compared. Among the identified proteins, differential parasitic markers with serodiagnostic potential, due to their well-known immunoreactivity in human, included Ag5, AgB proteins, 8-kDa glycoproteins, hydatid disease diagnostic antigen P29 and major egg antigen P40. In particular, seven proteoforms of AgB and 8-kDa glycoprotein resulted to be the most promising diagnostic biomarkers, as they might predict CE in ovine and discriminate between different types of parasites.

Molecular characterization and phylogenetic studies of Echinococcus granulosus and Taenia multiceps coenurus cysts in slaughtered sheep in Saudi Arabia.

Taeniids, consisting of two genera Echinococcus and Taenia, are obligatory tapeworms of mammals, and their pathogenicity was due to infection with larval stages. Hydatid (the larval stage of Echinococcus granulosus) and coenurus (the larval stage of Taenia multiceps) cysts are prevalent in domestic, wild ruminants, livestock, swine, and dogs, and accidentally they could also be found in humans. They lead to different clinical manifestations that cause economic loss in livestock and human morbidity. In Saudi Arabia, few studies were performed on hydatid and coenurus cyst genetic variations. The main goal of the present study was to identify E. granulosus and T. multiceps cyst isolates collected from slaughtered Harri sheep in Saudi Arabia by partial sequencing with PCR amplification of the cytochrome C oxidase 1 (COX1) gene. Molecular and phylogenetic evaluation based on COX1 sequences indicated that cyst isolates belong to E. granulosus and T. multiceps, respectively, successfully submitted in NCBI Genbank. Molecular characterization showed a low nucleotide diversity with two submitted isolates of coenurus with related isolates of Genbank. Conversely, E. granulosus isolates showed higher nucleotide diversity. The reported data could serve as a foundation for future molecular epidemiological and biological studies.

Comparative Transcriptome Analyses of the Developmental Stages of Taenia multiceps.

Cerebral coenurosis, caused by the larvae of Taenia multiceps (Coenurus cerebralis), is a fatal central nervous system disease in sheep and other herbivores and occasionally humans. Comparative transcriptomic profiles of the developmental stages of the parasite remain unknown. In this study, RNA sequencing was used to determine the transcriptome profiles of different stages of the life cycle of T. multiceps, including Oncosphere, Coenurus cerebralis (Pro with Cyst), and Adult (Adu), as well as scolex-neck proglottids (Snp), immature-mature proglottids (Imp), and gravid proglottids (Grp) of the adult stage. A total of 42.6 Gb (average 6.1 Gb) Illumina pair-end reads with a 125-bp read length were generated for seven samples. The total number of differentially expressed genes (DEGs) in the various life stages ranged from 2,577 to 3,879; however, for the tissues of the adult worm, the range was from 1,229 to 1,939. Kyoto Encyclopedia of Genes and Genomes analysis showed that the DEGs mainly participated in cellular and metabolic processes, binding and catalytic activity, genetic information processing, and environmental information processing. In addition, a large number of genes related to development and parasite-host interaction were identified. Quantitative reverse transcription-polymerase chain reaction confirmed that the levels of 28 selected DEGs were consistent with those determined using RNA sequencing. The present study provides insights into the mechanisms of the development and parasitic life of T. multiceps.

Cloning, expression and serodiagnostic potential of HSP70 of Taenia multiceps in sheep.

The aim of this study was to determine the efficacy of the recombinant protein expressed by the T. multiceps HSP70 gene in the immunodiagnosis of sheep coenurosis. Specific primers were designed to amplify the gene encoding HSP70 from the whole genome sequence of T. multiceps, mRNA was isolated from C. cerebralis cysts obtained from an infected sheep's brain, and then, cDNA was isolated. The gene region related to the designed primers was amplified by PCR, and the obtained product was cloned and expressed. The specificity of the recombinant protein purified afterwards was demonstrated by Western Blot using known positive and negative sheep sera. Additionally, the efficiency of the recombinant protein in ELISA was determined with sera collected from 1207 sheep. Finally, the sensitivity and specificity values ​of the recombinant TmHSP70 antigen in the Western Blot were 100 %, while the sensitivity and specificity rates of the ELISA were 66.6 % and 52.6 %, respectively. At the same time, 567 (46.97 %) of the 1207 analyzed serum samples were found to be positive by ELISA. In conclusion, the Western Blot technique had a higher quality than ELISA in detecting the TmHSP70 antigen for the serodiagnosis of sheep coenurosis.

First molecular confirmation of Coenurus cerebralis in sheep and goats with neurological behaviors in Iraq.

BACKGROUND AND AIM: Coenurus cerebralis is the intermediate stage of the canine cestode, which infect sheep and goats, resulting mainly in neurological signs and causing direct and indirect economic losses. This study aimed to demonstrate the existence of C. cerebralis and to elucidate the role of this parasite in inducing neurological behaviors in sheep and goats. MATERIALS AND METHODS: On the basis of historical data on neurological signs, we subjected 76 animals (49 sheep and 27 goats) of different ages, sexes, and geographical areas for molecular examination of their blood samples using the polymerase chain reaction assay. RESULTS: Of the 76 animals, 23.68% tested positive for C. cerebralis infection. We found significant increases in infection (p<0.05) in sheep (26.53%) more than in goats (18.52%). Circling movement was prevalent significantly among both C. cerebralis-positive sheep and goats. The Nineveh region had a significant (p<0.05) increase in positive sheep and goats, and the sheep of all study regions were infected significantly (p<0.05) more than were the goats. We found no significant (p>0.05) variation between sheep ages ≥1-3 years and ≥3 years; however, both groups had a significantly (p≤0.043) higher positivity rate than did sheep ages <1 year. The findings of sheep ages <1 year and ≥1-3 years were significantly higher than those of the goats, but not for goats ages ≥3 years. Female sheep and goats showed a significant increase in positivity versus that for the males. CONCLUSION: To the best of our knowledge, this study is the first report in Iraq targeting detection of C. cerebralis in sheep and goats with neurological behaviors; therefore, additional studies involving different animals in other regions using molecular techniques are needed.

Occurrence of Echinococcusgranulosussensulato and Other Taeniids in Bhutan.

The present research shows the results of a national study documenting the occurrence and genetic diversity of Echinococcus and Taenia species across Bhutan. Environmental dog faecal samples (n = 953) were collected from 2016 to 2018 in all 20 Bhutanese districts, mainly in urbanised areas. Cystic echinococcosis cysts were isolated from 13 humans and one mithun (Bos frontalis). Isolation of taeniid eggs from faeces was performed by sieving/flotation technique, followed by DNA isolation, PCR and sequence analyses for species identification (gene target: small subunit of ribosomal RNA). Genetic diversity of E. granulosuss.s. was based on the sequence (1609 bp) of the cox1 gene. A total of 67 out of 953 (7%) dog faecal samples were positive for at least one taeniid species. From the 670 free-roaming dog faecal samples, 40 (5.9%) were positive for taeniid DNA, 22 (3.2%) of them were identified as E. granulosuss.s. and four (0.5%) as E. ortleppi (G5). From the 283 faecal samples originating from yak-grazing areas, 27 (9.5%) were taeniid positive, including eight (2.8%) infected with E. granulosuss.s. and four (1.4%) with E. ortleppi. E. granulosuss.s. was identified in all isolates from human and the cyst from mithun. A haplotype network (cox1 gene) from E. granulosuss.s, including isolates from 12 dogs, two human and one mithun, revealed eight different haplotypes. The most common cox1 haplotype was the globally distributed Eg01, followed by Eg40 and Eg37 (previously described in China). Five new cox1 haplotypes (EgBhu1-5) originated from human, dogs, and a mithun were identified. The study indicated the contamination of urban areas and pastures with Echinococcus eggs in seven districts in Bhutan. The molecular characterisation of E. granulosuss.l. revealed different E. granulosuss.s. haplotypes as well as E. ortleppi. The transmission of T. multiceps was documented only in the western part of the country. Considering the zoonotic feature of E. granulosus s.s. and E. ortleppi and the economic impact of coenurosis caused by T. multiceps (also known as gid) in Bhutan, the findings of this study represent a significant contribution towards an epidemiological baseline for the establishment of a national control programme.

Are current practices of yak herdsmen adequate to combat Coenurosis in Laya Bhutan?

BACKGROUND: Coenurosis, known commonly as gid, is caused by a tapeworm Taenia multiceps. It is a disease of small ruminants globally but also occurs in large ruminants, especially in yak in the Himalaya. Gid is a pathological condition in young yaks, mostly below 3 years of age. The infected animal displays a circling movement with the head tilting towards the location of the cyst of a tapeworm on the cerebral surface of the brain. OBJECTIVE: We conducted a study with the primary aim to gain an insight into yak herdsmen's practices to manage Coenurosis in the Laya administrative block of Bhutan. METHODS: All seven villages of Laya were included for sampling. Seventy-five out of 182 households owned yaks, and 54 yak-owning households were selected randomly. The government livestock officials of nine yak-rearing highland districts were also included in the study. A cross-sectional study was conducted based on a questionnaire survey and focussed mainly on herdsmen's practices to manage gid. Two sets of questionnaires were used for yak herders and livestock officials. Each predesigned questionnaire was semi-structured and consisted of both open- and closed-ended questions. RESULTS: The study revealed gid as a major cause of yak mortality. Gid occurred more in winter during migration and most herders lost one to three yaks annually. Herdsmen kept an average of two watchdogs and dewormed them once annually. Similarly, calves were also dewormed once annually. The carcasses of dead yaks were fed to dogs. Livestock officials were optimistic about controlling the disease in the future, despite the yak areas being difficult to access. Most herders had not attended the gid awareness programme. The animal health worker visited herds once annually. In absence of animal health workers, most herdsmen resorted to different practices to treat affected yaks -the most common practice being surgery. Gid was mentioned to harm herdsmen's economy. CONCLUSIONS: The study concluded that for effective management of gid in Laya, livestock agencies must create more awareness on gid, increase the frequency of visits by animal health workers to yak herds, and increase the frequency of deworming of watchdogs and calves.

Cerebral Coenurosis of a Long-Tailed Goral, Naemorhedus caudatus, in Korea.

We intended to describe a case of cerebral coenurosis in a long-tailed goral, Naemorhedus caudatus, from Hwacheon-gun, Gangwon-do (Province), in the Korea. The goral, a 10-year-old male, was suffering from neurological symptoms, such as turning the circle to one side without lifting the head straight, and died at 30 days after admission to the wildlife medical rescue center in Chuncheon-si, Gangwon-do. A fluid-filled cyst was detected in the left cerebral hemisphere by computed tomography and magnetic resonance imaging. The cyst removed from the deceased goral was transparent, about 3×3 cm in size, contained a clear fluid and approximately 320 protoscolices invaginating from the internal germinal layer. The protoscolex had 4 suckers and a rostellum with 28 hooklets arranged in 2 rows. By the present study, a case of cerebral coenurosis was first confirmed in a long-tailed goral, N. caudatus, from Gangwon-do, in Korea. The residents frequently exposed in the sylvatic environment should be careful the accidental infections of zoonotic metacestode of Taenia multiceps, Coenurus cerebralis, in Korea.

First Report on Molecular Characterization of Taenia multiceps Isolates From Sheep and Goats in Faisalabad, Pakistan.

Coenurus cerebralis is the larval stage of Taenia multiceps commonly found in the brain (cerebral form), intramuscular and subcutaneous tissues (non-cerebral form) of ungulates. Globally, few reports exist on the molecular characterization and genetic diversity of C. cerebralis with none available for Pakistan. The current study molecularly characterized 12 C. cerebralis isolates surgically recovered from sheep (n = 4) and goats (n = 8) from a total of 3,040 small ruminants using a portion of the cytochrome c oxidase subunit 1 (cox1) mitochondrial (mt) gene. NCBI BLAST search confirmed the identity of each isolate. A high haplotype and a low nucleotide diversity with three haplotypes from the 12 isolates were observed. The findings suggest the existence of unique haplotypes of C. cerebralis in Pakistan. The negative value of Tajima's D and the positive value of Fu's Fs were inconsistent with population expansion, however, the sample size was small. Bayesian phylogeny revealed that all Pakistani isolates alongside the Chinese sequences (obtained from GenBank) constituted a cluster while sequences from other regions constituted another cluster. This is the first molecular study to determine the genetic diversity of C. cerebralis in Pakistan and serves as a foundation for prospective studies on the prevalence and population structure of C. cerebralis in the country. Furthermore, in this study, we amplified only a partial segment of the cox1 gene from a limited sample size. This could have implications on the interpretation of the actual population structure in reality. Thus, we recommend future studies to consider a larger sample size in a massive epidemiological survey for further insights.

In Silico Analysis of the Biodiversity and Conservation Status of Mitochondrial Cytochrome C Oxidase Subunit 1 (CO1) Gene of Taenia multiceps.

PURPOSE: Taenia multiceps resides in the small intestine of carnivores such as dogs, foxes, woles, jackals, while Coenurus cerebralis which is the larval form usually settle in the central nervous system and spinal cord of intermediate hosts like sheep, cattle and goats. The aim of this study was to analyze the haplotype diversity, genetic variation and population structure of the mt-CO1 gene sequences of sheep isolates of T. multiceps had been submitted to GenBank from different countries. METHODS: A total of 102 sequences from the mt-CO1 gene fragment belong to T. multiceps sheep isolates were used for bioinformatic analyses. Haplotype analysis, phylogenetic analysis and diversity, neutrality, fixation and gene flow analyses were applied to the sequences. RESULTS: As a result, 20 haplotypes together with different multiple nucleotide changes were determined after the sequence analysis. Trimmed fragment length was 337 bp hereby 19 polymorphic areas, 12 of which were parsimony informative, were identified, and any insertion-deletion was found. The number of mutations between major haplotypes and the others range from one to nine. The highest (0.72) genetic differentiation (Fst value) was observed between Turkey and Egypt populations while the lowest (- 0.22) was reported from Greece. These findings are important in terms of showing the diversity of nucleotide variation in T. multiceps sheep isolates. CONCLUSIONS: This study serves as the basis for future large-scale studies on T. multiceps worldwide epidemiology, bioecology, geographic distribution and population structure.

Coenurosis of Yak, Bos grunniens, caused by Taenia multiceps: A Case Report with Molecular Identification in Qinghai Tibetan Plateau Area, China.

Coenurosis is an important zoonotic helminthic disease caused by the larval stage of the tapeworm Taenia multiceps. This parasite typically infects the brain of the intermediate hosts, including sheep, goat, cattle and even humans. We report a case of T. multiceps infection in a yak confirmed by clinical symptoms, morphological characteristics, and molecular and phylogenetic analyses. The coenurus was thin-walled, whitish, and spherical in shape with a diameter of 10 cm. The parasite species was identified as T. multiceps by PCR amplification and sequencing of the 18S rRNA, cox1 and nad1 genes. Three gene sequences all showed high homology (all above 97%) with the reference sequences from different hosts. Moreover, phylogenetic reconstructions with the 3 published Taenia gene sequences confirmed that the Qinghai yak isolate was closely related to T. multiceps. Although there are advanced diagnosis and treatment methods for coenurosis, early infection is difficult to diagnose. Importantly, the findings of yak infection case should not be ignored due to its zoonotic potential.

Expression and serodiagnostic potential of antigen B and thioredoxin peroxidase from Taenia multiceps.

Coenurosis is a serious parasitic disease of herbivorous animals caused by the metacestode of Taenia multiceps (Coenurus cerebralis). Accordingly, a significant amount of research is currently dedicated to the development of appropriate antigens for use in rapid and accurate coenurosis diagnosis kits. In the present study, antigen B (AgB) and thioredoxin peroxidase (TPx) from T. multiceps were cloned and expressed using a prokaryotic system, molecular characterization of Tm-AgB was determined by bioinformatical analyses. The serological diagnostic potentials of rTm-AgB and rTm-TPx were evaluated by indirect ELISA and compared with those of previously reported rTm-AnxB2, rTm-HSP70, and rTm-GST. The results showed that Tm-AgB is a specific lipoprotein of cestodes with good thermal stability. The ELISA assay showed that rTm-AgB exhibited a sensitivity of 95.8% and a specificity of 87.5%, indicating its strong potential for serological diagnosis of T. multiceps.

A report on Coenuruses cerebralis infection in a wild goat (Capra aegagrus).

Coenurus cerebralis is the larval stage of Taenia multiceps inhabiting the small intestine of dogs and wild carnivores as the definitive hosts. A two-year-old wild female goat (Capra aegagrus) was referred with signs of lateral recumbency and seizure for four days and loss of appetite. In clinical examination, paddling, convulsion, and unconsciousness were observed indicating central nervous system disorder. Biochemical analyses showed increased levels of hematocrit, creatinine phosphorous, total bilirubin, direct bilirubin, blood urea nitrogen and calcium. No bacteria has been grown on culture medium taken from cerebrospinal fluid (CSF). The amount of total protein of the CSF was 1.10 g dL-1 (normal range = 20 - 40 mg dL-1). Hematological changes represented a left shift degenerative leukocytosis. At necropsy, two cysts sized over the 3 × 3 cm were detected, one on occipital lobe of the right hemisphere and the other on superior colliculi. The cysts contained a translucent fluid with a number of clusters of scolice growing from the inner layer of the cysts. To the best of our knowledge, this is the first report of coenurosis occurrence in Capra aegagrus.

Histopathological and Molecular Evaluation of the Experimentally Infected Goats by the Larval Forms of Taenia multiceps.

BACKGROUND: Introduction of Taenia multiceps and T. gaigeri as two separate species have been recognized mainly on morphological grounds. This experimental study was conducted to test whether cerebral and non-cerebral forms of Coenurus cerebralis belong to one origin or they are originated from two different tape worms. METHODS: Two groups of dogs were infected with the cerebral and muscular sources of the coenuri cysts. About two months later the eggs were collected from the fecal samples to be used to experimentally infect other healthy goats. Histopathological and molecular evaluation was conducted in two groups of goats that were challenged with T. multiceps eggs obtained from the infected dogs by brain and muscular sources of coenuri cysts in School of Veterinary Medicine of Shiraz University, Shiraz, Iran in 2015. All aberrant sites of predilection of the metacestode in goats were muscles, heart, diaphragm and lungs. The brain and spinal cord were carefully dissected and examined but the cysts were not found in these locations. In addition, the molecular genetic markers of mitochondrial DNA (CO1 and ND1) were applied to resolve the questionable relationship between T. multiceps and T. gaigeri. RESULTS: The larval stages of T. multiceps in brain and in other aberrant sites, which showed similar morphological criteria, were monophyletic species. CONCLUSION: Therefore, T. gaigeri must be considered taxonomically invalid.

Molecular characterisation and expression analysis of two heat-shock proteins in Taenia multiceps.

BACKGROUND: Taenia multiceps is a harmful tapeworm and its larval form (coenurus cerebralis) is the causative agent of coenurosis, a disease affecting the health of herbivores, resulting in great economic loss to animal husbandry. Heat-shock proteins (HSPs), expressed in all prokaryotes and eukaryotes, act as molecular chaperones and can affect pathogenicity. METHODS: Herein, cDNAs of T. multiceps genes Tm-HSP60 and Tm-p36 were cloned and molecularly characterised by bioinformatics analyses. The immunogenicity and immunoreactivity of recombinant rTm-HSP60 and rTm-p36 proteins were investigated by immunoblotting and indirect ELISA was established to evaluate their serodiagnostic potential. Tissue localisation and transcriptional level at different life stages of T. multiceps were determined by immunohistochemical and quantitative real-time PCR analyses. RESULT: The 533 residue rTm-HSP60 and the 314 residue rTm-p36 proteins share typical highly conserved features of HSPs. Tm-p36 shares structural characteristics with metazoan small HSPs, with two N-terminal α-crystallin domains. Compared with Tm-p36, Tm-HSP60 displayed stronger immunogenicity, and the indirect ELISA based on rTm-HSP60 exhibited a sensitivity of 83.3% and a specificity of 87.5%, while rTm-p36 was not suitable to develop indirect ELISA. Tm-HSP60 was widely distributed in all stages of T. multiceps, albeit at relatively low levels, while Tm-p36 was specifically distributed in the protoscolex and oncosphere. CONCLUSIONS: The sequence, structural and functional analyses of these two HSPs indicates that they may play important roles in the life-cycle of T. multiceps as molecular chaperones. Tm-HSP60 displayed stronger immunogenicity compare to Tm-p36, and has the potential for antibody detection. Tm-p36 was strongly associated with the activation of oncospheres and has potential interest for vaccination.

Coenurosis of Yak, Bos grunniens, caused by Taenia multiceps: A Case Report with Molecular Identification in Qinghai Tibetan Plateau Area, China

Coenurosis is an important zoonotic helminthic disease caused by the larval stage of the tapeworm Taenia multiceps. This parasite typically infects the brain of the intermediate hosts, including sheep, goat, cattle and even humans. We report a case of T. multiceps infection in a yak confirmed by clinical symptoms, morphological characteristics, and molecular and phylogenetic analyses. The coenurus was thin-walled, whitish, and spherical in shape with a diameter of 10 cm. The parasite species was identified as T. multiceps by PCR amplification and sequencing of the 18S rRNA, cox1 and nad1 genes. Three gene sequences all showed high homology (all above 97%) with the reference sequences from different hosts. Moreover, phylogenetic reconstructions with the 3 published Taenia gene sequences confirmed that the Qinghai yak isolate was closely related to T. multiceps. Although there are advanced diagnosis and treatment methods for coenurosis, early infection is difficult to diagnose. Importantly, the findings of yak infection case should not be ignored due to its zoonotic potential.

Molecular Characterization of Annexin B2, B3 and B12 in Taenia multiceps.

Coenurus cerebralis, the metacestode of Taenia multiceps, causes coenurosis, a disease severely affecting goat, sheep, cattle and yak farming and resulting in huge economic losses annually. Annexins bind calcium ions and play an important role in flatworm parasite development. To explore potential functions of annexins in T. multiceps, three homologous genes, namely, TmAnxB2, TmAnxB3 and TmAnxB12, were screened from the transcriptome dataset, amplified from C. cerebralis cDNA and subjected to bioinformatics analysis. Then, polyclonal antibodies recognizing the recombinant TmAnxB2 (rTmAnxB2) and rTmAnxB3 were prepared for localization of TmAnxB2 and TmAnxB3 in different tissues and developmental stages by immunofluorescence. The transcription of all three genes was also measured by relative fluorescent quantitative PCR. The sizes of rTmAnxB2, rTmAnxB3 and rTmAnxB12 were 58.00, 53.06 and 53.51 kDa, respectively, and rTmAnxB12 was unstable. Both rTmAnxB2 and rTmAnxB3 were recognized by goat-positive T. multiceps sera in Western blots. Immunofluorescence revealed that TmAnxB2 and TmAnxB3 were localized in the protoscolex and cyst wall and TmAnxB3 was also detected in adult cortex. TmAnxB2 and TmAnxB12 mRNA levels were determined to be highest in oncospheres and protoscolex, whereas transcription of TmAnxB3 was highest in scolex and immature segments. Taken together, these findings indicate that TmAnxB2 and TmAnxB12 may play critical roles in T. multiceps larvae, while TmAnxB3 may have important functions in adults. These results will lay the foundation for functional research of annexins in T. multiceps.