Four Human Cases of Diphyllobothrium nihonkaiense (Eucestoda: Diphyllobothriidae) in China with a Brief Review of Chinese Cases.

We described 4 human infection cases of zoonotic fish-tapeworm, Diphyllobothrium nihonkaiense, identified with morphological and molecular characters and briefly reviewed Chinese cases in consideration of it as an emerging parasitic disease in China. The scolex and mature and gravid proglottids of some cases were seen, a rosette-shaped uterus was observed in the middle of the mature and gravid proglottids, and the diphyllobothriid eggs were yellowish-brown in color and displayed a small knob or abopercular protuberance on the opposite end of a lid-like opening. The average size of the eggs was recorded as 62-67×42-45 µm. The parasitic materials gathered from 4 human cases were morphologically identified as belonging to the genera Diphyllobothrium and Adenocephalus. The phylogenetic analysis based on the nucleotide sequences of cytochrome c oxidase subunit 1 gene of the etiologic agents confirmed that the 4 cases were D. nihonkaiense infection. The finding of 4 additional D. nihonkaiense cases suggests that D. nihonkaiense might be a major causative species of human diphyllobothriasis in China. A combined morphological and molecular analysis is the main method to confirm D. nihonkaiense infection.

Molecular Identification of Diphyllobothrium latum from a Pediatric Case in Taiwan.

Human diphyllobothriasis is a parasitic disease caused by ingestion of larvae (plerocercoids) in raw or undercooked fish and commonly found in temperate areas. Rare cases were reported in tropical or subtropical areas especially in children. The first documented case of pediatric diphyllobothriasis in Taiwan had been reported 11 years ago. Here, we report another 8-year-old girl case who presented with a live noodle-like worm hanging down from her anus, with no other detectable symptoms. We pulled the worm out and found the strobila being 260 cm in length. Examination of gravid proglottids showed that they were wider than their lengths, containing an ovoid cirrus sac in the anterior side and the rosette-shaped uterus. Eggs extracted from the uterus were ovoid and operculated. Diphyllobothrium latum was confirmed by molecular analysis of the mitochondrial DNA cytochrome c oxidase subunit 1 (cox1) gene. The girl was treated with a single oral dose of praziquantel, and no eggs or proglottids were observed from her stool in the subsequent 3 months. The reemergence of human diphyllobothriasis in non-endemic countries is probably due to prevalent habit of eating imported raw fish from endemic areas. This pediatric case raised our concern that human diphyllobothriasis is likely underestimated because of unremarkable symptoms.

Four human cases of Diphyllobothrium latum infection.

Diphyllobothrium latum infections in 4 young Korean men detected from 2008 to 2012 are presented. Three were diagnosed based on spontaneously discharged strobila of the adult worm in their feces, and 1 case was diagnosed by finding the worm at colonoscopy examination in a local clinic. The morphologic characteristics of the gravid proglottid and eggs were consistent with D. latum. All patients were treated with praziquantel 15 mg/kg, and follow-up stool examinations were done at 2 months after the medication. The main clinical complaints were intermittent gastrointestinal troubles such as indigestion, abdominal distension, and spontaneous discharge of tapeworm's segments in their feces. The most probable source of infection was the flesh of salmon or trout according to a patient's past history. These are the 45th to 48th recorded cases diagnosed by the adult worm in the Republic of Korea since 1971.

Molecular and morphological identification of Adenocephalus pacificus (Cestoda) isolated from South American sea lion Otaria byronia stranded on the northern Peruvian coasts.

The most frequent etiologic agent of diphyllobothriosis in South America and the only one confirmed by molecular data in human cases in Peru is Adenocephalus pacificus (syn. Diphyllobothrium pacificum). This cestode is transmitted by ingestion of the plerocercoids found in marine fish, causing a parasitic zoonosis. The objective of the present study was to identify two cestodes isolated from two specimens of the South American sea lion (Otaria byronia) stranded on the beaches of Huacho and Barranca cities, located on the northern Peruvian coasts, in the department of Lima. Tapeworms were confirmed by morphological characteristics due to the presence of transverse papilla-like tegumental protuberances in proglottids and small sized eggs, as well as by sequencing of the partial cytochrome c oxidase subunit 1 (mtDNA-COI) gene that are congruent with additional available A. pacificus sequences. Even though sea lions in Peru are distributed along the coast and in areas of difficult access, generally located in protected natural areas, the fortuitous finding represented an opportunity to confirm the presence of A. pacificus in South American sea lions. This report of tapeworm A. pacificus could allow future monitoring of the occurrence and geographical distribution of this causative agent in epidemiological studies, since it is one of the main species of zoonotic importance in Peru.

Diphyllobothrium sprakeri n. sp. (Cestoda: Diphyllobothriidae): a hidden broad tapeworm from sea lions off North and South America.

BACKGROUND: The systematic of several marine diphyllobothriid tapeworms of pinnipeds has been revised in recent years. However, 20 species of Diphyllobothrium from phocids and otariids are still recognized as incertae sedis. We describe a new species of Diphyllobothrium from the intestine of California sea lions Zalophus californianus (Lesson) (type-host) and South American sea lions Otaria flavescens (Shaw). METHODS: Zalophus californianus from the Pacific coast of the USA and O. flavescens from Peru and Argentina were screened for parasites. Partial fragments of the large ribosomal subunit gene (lsrDNA) and the cytochrome c oxidase subunit 1 (cox1) mitochondrial gene were amplified for 22 isolates. Properly fixed material from California sea lions was examined using light and scanning electron microscopy. RESULTS: A total of four lsrDNA and 21 cox1 sequences were generated and aligned with published sequences of other diphyllobothriid taxa. Based on cox1 sequences, four diphyllobothriid tapeworms from O. flavescens in Peru were found to be conspecific with Adenocephalus pacificus Nybelin, 1931. The other newly generated sequences fall into a well-supported clade with sequences of a putative new species previously identified as Diphyllobothrium sp. 1. from Z. californianus and O. flavescens. A new species, Diphyllobothrium sprakeri n. sp., is proposed for tapeworms of this clade. CONCLUSIONS: Diphyllobothrium sprakeri n. sp. is the first diphyllobothriid species described from Z. californianus from the Pacific coast of North America, but O. flavescens from Argentina, Chile and Peru was confirmed as an additional host. The present study molecularly confirmed the first coinfection of two diphyllobothriid species in sea lions from the Southern Hemisphere.

Molecular identification of the Diphyllobothrium species causing diphyllobothriasis in Chilean patients.

Diphyllobothriasis caused by the infection of adult Diphyllobothrium tapeworms sporadically occurs in Chile. The occurrence of the disease is closely linked to the consumption of raw or undercooked freshwater and marine fishes. Diagnosis of diphyllobothriasis has been based on laboratory examinations of the morphological characteristics of proglottids and eggs passed in the feces. Although determination of the parasite to the species level is possible through histologic examination of proglottid specimens, the parasites of patients who only discharge eggs cannot be diagnosed to the species level. Determining the species responsible for the infection of humans and other animals in affected areas is an important component of understanding the epidemiologic and enzootic characteristics of any infectious disease. We therefore compared the classification results obtained using a molecular approach with those obtained from morphological and histopathological examination of proglottids or eggs from five Chilean individuals with diphyllobothriasis. DNA analysis confirmed that the causative Diphyllobothrium species in Chile were first identified as Diphyllobothrium latum and Diphyllobothrium pacificum at least. Furthermore, mitochondrial cytochrome c oxidase subunit 1 gene analysis also supported the hypothesis that D. latum from Chile originated from Europe.

Morphologic and genetic identification of Diphyllobothrium nihonkaiense in Korea.

Diphyllobothrium nihonkaiense was first described by Yamane in 1986 but the taxonomical features have been obscure due to lack of critical morphologic criteria in its larval and adult stages. In Korea, this tapeworm had long been known as Diphyllobothrium latum. In this study, we observed 62 specimens collected from Korean residents and analyzed them by morphological features and nucleotide sequences of mitochondrial cox1 gene as well as the ITS1 region. Adult tapeworms were examined after carmine or trichrome stain. Longitudinal sections of the gravid proglottids showed an obtuse angle of about 150 degree between the cirrus sac and seminal vesicle. This angle is known as a major differential point compared with that of D. latum. Nucleotide sequence differences between D. latum and the specimens from Koreans represented 17.3% in mitochondrial DNA cox1 gene. Sequence divergence of ITS1 among 4 Korean isolates was 0.3% and similarity was 99.7% with D. nihonkaiense and D. klebanovskii. All of the Korean specimens analyzed in this study were identified as being D. nihonkaiense (n = 62). We propose its Korean name as "Dong-hae-gin-chon-chung" which means 'long tapeworm of the East Sea' for this newly analyzed diphyllobothriid tapeworm in Korea.

Complex insight on microanatomy of larval "human broad tapeworm" Dibothriocephalus latus (Cestoda: Diphyllobothriidea).

BACKGROUND: In Europe, the tapeworm Dibothriocephalus latus (syn. Diphyllobothrium latum) is a well-known etiological agent of human diphyllobothriosis, which spreads by the consumption of raw fish flesh infected by plerocercoids (tapeworm's larval stage). However, the process of parasite establishment in both intermediate and definitive hosts is poorly understood. This study was targeted mainly on the scolex (anterior part) of the plerocercoid of this species, which facilitates penetration of the parasite in intermediate paratenic fish hosts, and subsequently its attachment to the intestine of the definitive host. METHODS: Plerocercoids were isolated from the musculature of European perch (Perca fluviatilis) caught in Italian alpine lakes. Parasites were examined using confocal microscopy, scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Immunofluorescence tagging was held on whole mount larvae. RESULTS: The organisation of the central and peripheral nervous system was captured in D. latus plerocercoids, including the ultrastructure of the nerve cells possessing large dense neurosecretory granules. Two types of nerve fibres run from the body surface toward the nerve plexus located in the parenchyma on each side of bothria. One type of these fibres was found to be serotoninergic and possessed large subtegumental nerve cell bodies. A well-developed gland apparatus, found throughout the plerocercoid parenchyma, produced heterogeneous granules with lucent core packed in a dense layer. Three different types of microtriches occurred on the scolex and body surface of plerocercoids of D. latus: (i) uncinate spinitriches; (ii) coniform spinitriches; and (iii) capilliform filitriches. Non-ciliated sensory receptors were observed between the distal cytoplasm of the tegument and the underlying musculature. CONCLUSIONS: Confocal laser scanning microscopy and electron microscopy (SEM and TEM) showed the detailed microanatomy of the nervous system in the scolex of plerocercoids, and also several differences in the larval stages compared with adult D. latus. These features, i.e. well-developed glandular system and massive hook-shaped uncinate spinitriches, are thus probably required for plerocercoids inhabiting fish hosts and also for their post-infection attachment in the human intestine.

A case of human diphyllobothriasis in northern Taiwan after eating raw fish fillets.

The consumption of raw fish fillets is increasing in Taiwan. A male Taiwanese aged 30 years presented after passing a flat, white noodle-like worm. Strobila examination showed that most proglottids were wider than they were long, with the genital pore located at the posterior edge of the cirrus. Histological and coprological findings confirmed the diagnosis of Diphyllobothrium latum; ova were ellipsoidal with operculate characteristics, and had a small knob in the anti-operculum side. Hematological data, including vitamin B12 levels, were normal, except for a low folate level. The patient was treated with a single dose of praziquantel 600 mg and 196 cm of proglottids were expelled during the 3 days following treatment. Further follow-up was declined. Consumption of raw and undercooked fish (especially salmon) poses a risk of D. latum infection.

Diphyllobothriasis in Saudi Arabia.

We described a Saudi patient infected with Diphyllobothrium latum D. latum. A 38-year-old male presented, complaining of passing worms. He had a history of recent travel to Europe and South East Asia. Stools examination revealed typical D. latum eggs. He was treated with praziquantel followed by saline purge, after which he discharged an intact tapeworm. Macroscopic and microscopic examinations of the worm confirmed the diagnosis of D. latum. This is the first case of diphyllobothriasis to be reported in Saudi Arabia. The epidemiology and methods of prevention of diphyllobothriasis are discussed.

Diphyllobothriasis latum: the first child case report in Taiwan.

Diphyllobothriasis latum is an intestinal parasitosis caused by the ingestion of mostly raw fresh-water fish containing plerocercoids of Diphyllobothrium latum. We report an 8-year-old boy who came to our hospital with the complaint of a tapeworm hanging from the anus after defecation. The other symptom was mild abdominal cramping for a period of 1 year. The laboratory examination did not reveal anemia or vitamin B12 deficiency. Examination of gravid proglottids with rosette-like central uterus and typically operculated eggs of D. latum confirmed the diagnosis. The morphologic characters of proglottids and eggs size are compatible with D. latum. The patient had a history of eating uncooked fish for 1 year. Salmonids may be the infection source. He was treated with two doses of praziquantel and passed about 183 cm in length of all proglottids. There is a high prevalence of diphyllobothriasis latum in the northern temperate areas, but it is very rare in children. This patient is the first child case reported in Taiwan.

Eggs as a Suitable Tool for Species Diagnosis of Causative Agents of Human Diphyllobothriosis (Cestoda).

BACKGROUND: Tapeworms of the order Diphyllobothriidea are parasites of tetrapods and several species may infect man and cause neglected human disease called diphyllobothriosis. Identification of human-infecting diphyllobothriid cestodes is difficult because of their morphological uniformity, which concerns also their eggs in stool samples. METHODS: In the present study, we analysed by far the largest dataset of more than 2,000 eggs of 8 species of diphyllobothriid cestodes that may infect humans, including the most frequent human parasites Diphyllobothrium latum, D. nihonkaiense and Adenocephalus pacificus (syn. Diphyllobothrium pacificum). Size (length, width and length/width ratio) and the surface of the egg shell from naturally and experimentally infected hosts were studied using light and scanning electron microscopy. RESULTS: A high degree of intraspecific and host-related size variability has been detected, but combination of morphometrical and ultrastructural data made it possible to distinguish all of the studied species, including otherwise quite similar eggs of the 3 most common species infecting man, i.e. D. latum, D. nihonkaiense and D. dendriticum. The surface of all marine species is covered by numerous deep pits with species-specific density, whereas the surface of freshwater species is smooth or with isolated shallow hollows or wrinkles.

High morphological plasticity and global geographical distribution of the Pacific broad tapeworm Adenocephalus pacificus (syn. Diphyllobothrium pacificum): molecular and morphological survey.

The most important causative agent of human diphyllobothriosis in South America, Diphyllobothrium pacificum, is transferred to the original genus Adenocephalus Nybelin, 1931; revised and redescribed on the basis of the evaluation of an extensive material collected mainly from northern fur seal, Callorhinus ursinus, from St. Paul Island, Alaska. Detailed analysis of morphological and morphometrical data shows a high variability in most of the characteristics traditionally used in diagnosis of diphyllobothriid tapeworms. Phylogenetic analyses based on newly characterised sequences of mitochondrial cytochrome c oxidase subunit 1 and nuclear large subunit ribosomal RNA genes consistently reveal Adenocephalus pacificus as a sister lineage to the clade formed of the remaining Diphyllobothrium species and other genera (Digramma, Diplogonoporus, Ligula). Despite the generally similar morphology, A. pacificus can be differentiated from the closely related taxa in the presence of transverse papilla-like tegumental protuberances distributed anteriorly, separated by narrow semicircular grooves on the ventral surface of proglottids between their anterior margin and the anterior edge of the male gonopore, and relatively small eggs. A. pacificus displays a relatively low host specificity (found in 9 of 16 otariids, and in accidental hosts such as man, dog and jackal, the latter representing a new host) and a uniquely wide geographical distribution on both hemispheres. In addition, suitability of morphological criteria used in diagnostics of diphyllobothriid cestodes is discussed.

Five cases of Diphyllobothrium nihonkaiense infection with discovery of plerocercoids from an infective source, Oncorhynchus masou ishikawae.

Five persons from 2 families residing at Miyama Town, Mie Prefecture, Japan, ingested fresh raw fish Oncorhynchus sp. on 9 May 1999 that was caught at Owase district in Mie. They all expelled diphyllobothriid cestodes 11-37 days after ingesting the fish. The parasites were morphologically identical to Diphyllobothrium nihonkaiense Yamane et al., 1986. Five plerocercoids were detected from a portion of the fish. Nucleotide sequence of a region of the cytochrome c oxidase subunit I gene of mitochondrial DNA from an adult worm was identical with that from the plerocercoid. The fish was identified as Oncorhynchus masou ishikawae according to the nucleotide sequence of the nuclear ribosomal second internal transcribed spacer region II gene. This is the first record of D. nihonkaiense plerocercoids from O. m. ishikawae.

Two rare cases of Diphyllobothrium latum parvum type infection in Korea.

Two rare cases of human infection with parvum (dwarf) type of Diphyllobothrium latum [syn. D. parvum (Stephens, 1908)], were discovered in Korea. The first case was a 46-year old housewife, from whom a kind of pseudophyllidean tapeworm eggs was detected in the feces. She was treated with praziquantel and purged, and a complete strobila with scolex, 120 cm in total length, was recovered. She recalled that she had eaten raw trouts at a raw-fish restaurant near the Chungju Lake. Another patient was a 22-year old medical student (male), who used to eat raw sea-foods. He discovered a chain of tapeworm proglottids, 15 cm in length, discharged spontaneously in his stool and brought it for identification. The worms from the two cases were compatible with D. parvum (Stephens, 1908) of which the taxonomic significance has long been questioned. After a detailed morphological study and review of literature, we designated the worms as D. latum parvum type. This is the first report on the occurrence of this rare type of D. latum infection in Korea.

New cases of human diphyllobothriasis in Patagonia, Argentina.

The purpose of the paper is to describe 4 new cases of human diphyllobothriasis in Patagonia, Argentine. Adult parasites recovered were submitted to morphological and histological analyses for taxonomic identification. The etiological agent found was always Diphyllobothrium latum and all the cases were autochthonous. These data combined with previous information make the number of autochthonous human cases of diphyllobothriosis registered in Argentina increase to 18.

[Anatomy of the neural apparatus of cestodes of the families Amphicotylidae and Diphyllobothriidae (Pseudophyllidea)]. / Anatomiia nervnogo apparata tsestod semeistv Amphicotylidae i Diphyllobothriidae (Pseudophyllidea).

Studies were carried out of the nervous apparatus of Eubothrium salvelini, E. rugosum, E. crassum, E. vittevitellatus, Diphyllobothrium latum and D. dendriticum (Cestoda, Pseudophyllidea) at different stages of their life cycle. Scolexes of adult cestodes are innervated with 5 pairs of longitudinal nerve trunks, the number of which in the strobile gradually increases up to 17 pairs in species of Eubothrium and up to 60 pairs (in the wildest parts of the strobile) in species of Diphyllobothrium. There were noted differences in of longitudinal trunks in different species and in the specimens of the same species but from different hosts, the width of the strobile being the same. The main lateral trunks are situated in the medullar parenchima. In species of Eubothrium they are wide apart while in Diphyllobothrium they are drawn closer to one another. The description of the rough and inner nerve plexuses is given. Judging by the larval developmental phases the well developed orthogon with 3 pairs of longitudinal nerve trunks can be taken for the initial state of the nervous apparatus in Pseudophyllidea. This means that the initial forms belonged to advanced turbellarians rather than to primitive ones.

[Diphyllobothrium lobodoni sp. n. (Cestoda: Diphyllobothriidae)--a parasite of the crabeater seal]. / Diphyllobothrium lobodoni sp. n. (Cestoda: Diphyllobothriidae)--parazit tiulenia-kraboeda.

The new cestode species is described from the intestine of the crab eater seal Lobodon carcinophagus living in Pacific region of Antarctic. It differs from other diphyllobothriids by clear wavy lateral margins of the body. It is close to D. scoticum by the presence of uterine sack, but it has larger size of the body, lesser craspedotity of segments, lesser size of scolex, longer neck, thinner tegument, 3 times thicker layers of longitudinal and transversal muscles, 2 times greater number of testicles (per transversal section of the segments), another form and position of the bursa of cirrus and of the testicular vesicle, larger uterine sack consisting about one third of the total uterine volume.

[Causative agent of human diphyllobothriasis--Diphyllobothrium klebanovskii sp. n]. / Vozbuditel' difillobotrioza cheloveka--Diphyllobothrium klebanovskii sp. n.

A new cestode species, Diphyllobothrium klebanovskii sp. n., is described on the basis of experimental examination and morphological study of 51 specimens expelled from men in lower part of the Amur river and 18 specimens expelled from golden hamsters. The larval stage of this cestode is plerocercoid of type F, which is localized in muscles of Oncorhynchus keta and O. gorbuscha. D. klebanovskii sp. n. is a marine species, it differs from D. latum and other species of the genus Diphyllobothrium.