Molecular identification of Spirometra infections in companion animals and wildlife in Japan.

Spirometra infections in companion animals and wildlife in Japan have been diagnosed based on the morphology of the adult worms and eggs, and the etiological agent has been mainly ascribed to Spirometra erinaceieuropaei. However, recent studies have revealed that two other species, Spirometra mansoni and Spirometra asiana, coexist in Japan. Spirometra asiana is a new species recently discovered in Japan. Although morphological discrimination between these two species is difficult, molecular identification is useful. Therefore, to understand which species commonly parasitizes companion animals and wildlife in Japan, a preliminary study was performed based on mitochondrial DNA analysis. Eleven adult worms examined were identified as S. mansoni, suggesting that S. mansoni infects companion animals and wildlife commonly than S. asiana in Japan.

Morphological and Molecular Identification of Spirometra Tapeworms (Cestoda: Diphyllobothriidae) from Carnivorous Mammals in the Serengeti and Selous Ecosystems of Tanzania.

Spirometra tapeworms (Cestoda: Diphyllobothriidae) collected from carnivorous mammals in Tanzania were identified by the DNA sequence analysis of the mitochondrial cytochrome c oxidase subunit 1 (cox1) and internal transcribed spacer 1 (ITS1), and by morphological characteristics. A total of 15 adult worms were collected from stool samples and carcasses of Panthera leo, Panthera pardus, and Crocuta crocuta in the Serengeti and Selous ecosystems of Tanzania. Three Spirometra species: S. theileri, S. ranarum and S. erinaceieuropaei were identified based on morphological features. Partial cox1 sequences (400 bp) of 10 specimens were revealed. Eight specimens showed 99.5% similarity with Spirometra theileri (MK955901), 1 specimen showed 99.5% similarity with the Korean S. erinaceieuropaei and 1 specimen had 99.5% similarity with Myanmar S. ranarum. Sequence homology estimates for the ITS1 region of S. theileri were 89.8% with S. erinaceieuropaei, 82.5% with S. decipiens, and 78.3% with S. ranarum; and 94.4% homology was observed between S. decipiens and S. ranarum. Phylogenetic analyses were performed with 4 species of Spirometra and 2 species of Dibothriocephalus (=Diphyllobothrium). By both ML and BI methods, cox1 and ITS1 gave well supported, congruent trees topology of S. erinaceieuropaei and S. theileri with S. decipiens and S. ranarum forming a clade. The Dibothriocephalus species were sisters of each other and collectively forming successive outgroups. Our findings confirmed that 3 Spirometra species (S. theileri, S. ranarum, and S. erinaceieuropaei) are distributed in the Serengeti and Selous ecosystems of Tanzania.

Molecular and Morphologic Identification of Spirometra ranarum Found in the Stool of African Lion, Panthera leo in the Serengeti Plain of Tanzania.

The present study was performed with morphological and molecular analysis (cox1 and nad1 mitochondrial genes) to identify the proglottids of spirometrid tapeworm found in the stool of an African lion, Panthera leo, in the Serengeti plain of Tanzania. A strand of tapeworm strobila, about 75 cm in length, was obtained in the stool of a male African lion in the Serengeti National Park (34˚ 50' E, 02˚ 30' S), Tanzania, in February 2012. The morphological features of the adult worm examined exhibited 3 uterine coils with a bow tie appearance and adopted a diagonal direction in the second turn. The posterior uterine coils are larger than terminal uterine ball and the feature of uteri are swirling rather than spirally coiling. The sequence difference between the Spirometra species (Tanzania origin) and S. erinaceieuropaei (GenBank no. KJ599680) was 9.4% while those of S. decipiens (GenBank no. KJ599679) differed by 2.1% in the cox1 and nad1 genes. Phylogenetic tree topologies generated using the 2 analytic methods were identical and presented high level of confidence values for the 3 major branches of the 3 Spirometra species in the cox1 gene. The morphological and molecular findings obtained in this study were nearly coincided with those of S. ranarum. Therefore, we can know for the first time that the African lion, Panthera leo, is to the definitive host of this tapeworm.

Genetic and Morphologic Identification of Spirometra ranarum in Myanmar.

In the present study, we identified a Spirometra species of Myanmar origin (plerocercoid) by molecular analysis using mitochondrial cox1 and nad1 genes, as well as by morphological observations of an adult tapeworm. Spargana specimens were collected from a paddy-field in Taik Kyi Township Tarkwa Village, Yangon, Myanmar in December 2017. A total of 5 spargana were obtained from 20 frogs Hoplobatrachus rugulosus; syn: Rana rugulosa (Wiegmann, 1834) or R. tigrina (Steindachner, 1867). The plerocercoids were used for experimental infection of a dog. After 4 weeks of infection, an adult tapeworm was recovered from the intestine of the dog. Morphologically, the distinct features of Spirometra sp. (Myanmar origin) relative to S. erinaceieuropaei and S. decipiens include a uterine morphology comprising posterior uterine coils that larger than the terminal uterine ball and coiling of the uteri diagonally (swirling) rather than spirally. The cox1 sequences (1,566 bp) of the Myanmar-origin Spirometra species showed 97.9% similarity to a reference sequence of S. decipiens (GenBank no. KJ599679) and 90.5% similarity to a reference sequence of S. erinaceieuropaei (GenBank no. KJ599680). Phylogenetic tree topologies were identical and presented high confidence level of values for the 3 major branches of the 3 Spirometra species in cox1 and nad1 genes. These results indicated that Myanmar-origin Spirometra species coincided with those of S. ranarum and may be considered as a valid species.

Spirometra decipiens (Cestoda: Diphyllobothriidae) Collected in A Heavily Infected Stray Cat from the Republic of Korea.

Morphological and molecular characteristics of spirometrid tapeworms, Spirometra decipiens, were studied, which were recovered from a heavily infected stray cat road-killed in Eumseong-gun, Chungcheongbuk-do (Province), the Republic of Korea (=Korea). A total of 134 scolices and many broken immature and mature proglottids of Spirometra tapeworms were collected from the small intestine of the cat. Morphological observations were based on 116 specimens. The scolex was 22.8-32.6 mm (27.4 mm in average) in length and small spoon-shape with 2 distinct bothria. The uterus was coiled 3-4 times, the end of the uterus was ball-shaped, and the vaginal aperture shaped as a crescent moon was closer to the cirrus aperture than to the uterine aperture. PCR amplification and direct sequencing of the cox1 target fragment (377 bp in length and corresponding to positions 769-1,146 bp of the cox1 gene) were performed using total genomic DNA extracted from 134 specimens. The cox1 sequences (377 bp) of the specimens showed 99.0% similarity to the reference sequence of S. decipiens and 89.3% similarity to the reference sequence of S. erinaceieuropaei. In the present study, we report a stray cat heavily infected with S. decipiens identified by mitochondrial cox1 sequence analysis and morphological examinations of the adult worms.

Molecular Detection of Spirometra decipiens in the United States.

The genus Spirometra belongs to the family Diphyllobothriidae and order Pseudophyllidea, and includes intestinal parasites of cats and dogs. In this study, a plerocercoid labeled as Spirometra mansonoides from the USA was examined for species identification and phylogenetic analysis using 2 complete mitochondrial genes, cytochrome c oxidase I (cox1) and NADH dehydrogenase subunit 3 (nad3). The cox1 sequences (1,566 bp) of the plerocercoid specimen (USA) showed 99.2% similarity to the reference sequences of the plerocercoid of Korean Spirometra decipiens (GenBank no. KJ599679), and 99.1% similarity in regard to nad3 (346 bp). Phylogenetic tree topologies generated using 4 analytical methods were identical and showed high confidence levels with bootstrap values of 1.00, 100%, 100%, and 100% for Bayesian inference (BI), maximum-likelihood (ML), neighbor-joining (NJ), and maximum parsimony (MP) methods, respectively. Representatives of Diphyllobothrium and Spirometra species formed a monophyletic group, and the sister-genera status between these species was well supported. Trapezoic proglottids in the posterior 1/5 region of an adult worm obtained from an experimentally infected cat were morphologically examined. The outer uterine loop of the uterus coiling characteristically consisted of 2 complete turns. The results clearly indicated that the examined Spirometra specimen from the USA matched to S. decipiens very well, and indicated possible presence of the life cycle of this species in this region.

[Stereotactic aspiration of Spirometra mansonides larvae]. / Aspiración estereotáctica de una larva de Spirometra spp.

Brain sparganosis is a non-common parasite infection by Diphyllobothrium or Spirometra mansonoides larvae. This last one is responsible for most of the infestations in humans. We report a 19 years male patient bearer of a brain sparganosis. The patient presented with headache and left hemiparesis. CT diagnosis of right thalamic lesions was made and aspiration biopsy was performed using stereotactic system, obtaining a whole and death larvae. Histopathology confirms a CNS parasitism and it was treated initially with albendazol. ELISA test confirmed Spirometra spp. infestation. The patient developed asymptomatic with total remission of the lesions. It constitutes the second report in Cuba of brain sparganosis.

Aspiración estereotáctica de una larva de Spirometra spp. / Stereotactic aspiration of Spirometra mansonides larvae

Brain sparganosis is a non-common parasite infection by Diphyllobothrium or Spirometra mansonoides larvae. This last one is responsible for most of the infestations in humans. We report a 19 years male patient bearer of a brain sparganosis. The patient presented with headache and left hemiparesis. CT diagnosis of right thalamic lesions was made and aspiration biopsy was performed using stereotactic system, obtaining a whole and death larvae. Histopathology confirms a CNS parasitism and it was treated initially with albendazol. ELISA test confirmed Spirometra spp. infestation. The patient developed asymptomatic with total remission of the lesions. It constitutes the second report in Cuba of brain sparganosis.

Se presenta el caso clínico de un varón con 19 años de edad y el diagnóstico de una esparganosis cerebral. Consultó por cefalea y una hemiparesia izquierda. En una tomografía computarizada cerebral con contraste se observaron lesiones talámicas derechas. Se realizó una biopsia cerebral guiada por estereotaxia con aspiración completa de un verme. En el estudio histopatológico se planteó un probable parasitismo de SNC y fue tratado inicialmente con albendazol. Se confirmó la infección por Spirometra spp. por test de ELISA. Evolucionó con regresión de síntomas y remisión imagenológica de las lesiones. Este caso constituye el segundo reporte en Cuba de una infestación cerebral por este parásito y aspiración estereotáctica de la larva de Spirometra spp.

Human Infections with Spirometra decipiens Plerocercoids Identified by Morphologic and Genetic Analyses in Korea.

Tapeworms of the genus Spirometra are pseudophyllidean cestodes endemic in Korea. At present, it is unclear which Spirometra species are responsible for causing human infections, and little information is available on the epidemiological profiles of Spirometra species infecting humans in Korea. Between 1979 and 2009, a total of 50 spargana from human patients and 2 adult specimens obtained from experimentally infected carnivorous animals were analyzed according to genetic and taxonomic criteria and classified as Spirometra erinaceieuropaei or Spirometra decipiens depending on the morphology. Morphologically, S. erinaceieuropaei and S. decipiens are different in that the spirally coiled uterus in S. erinaceieuropaei has 5-7 complete coils, while in S. decipiens it has only 4.5 coils. In addition, there is a 9.3% (146/1,566) sequence different between S. erinaceieuropaei and S. decipiens in the cox1 gene. Partial cox1 sequences (390 bp) from 35 Korean isolates showed 99.4% (388/390) similarity with the reference sequence of S. erinaceieuropaei from Korea (G1724; GenBank KJ599680) and an additional 15 Korean isolates revealed 99.2% (387/390) similarity with the reference sequences of S. decipiens from Korea (G1657; GenBank KJ599679). Based on morphologic and molecular databases, the estimated population ratio of S. erinaceieuropaei to S. decipiens was 35: 15. Our results indicate that both S. erinaceieuropaei and S. decipiens found in Korea infect humans, with S. erinaceieuropaei being 2 times more prevalent than S. decipiens. This study is the first to report human sparganosis caused by S. decipiens in humans in Korea.

Human Infections with Spirometra decipiens Plerocercoids Identified by Morphologic and Genetic Analyses in Korea

Tapeworms of the genus Spirometra are pseudophyllidean cestodes endemic in Korea. At present, it is unclear which Spirometra species are responsible for causing human infections, and little information is available on the epidemiological profiles of Spirometra species infecting humans in Korea. Between 1979 and 2009, a total of 50 spargana from human patients and 2 adult specimens obtained from experimentally infected carnivorous animals were analyzed according to genetic and taxonomic criteria and classified as Spirometra erinaceieuropaei or Spirometra decipiens depending on the morphology. Morphologically, S. erinaceieuropaei and S. decipiens are different in that the spirally coiled uterus in S. erinaceieuropaei has 5-7 complete coils, while in S. decipiens it has only 4.5 coils. In addition, there is a 9.3% (146/1,566) sequence different between S. erinaceieuropaei and S. decipiens in the cox1 gene. Partial cox1 sequences (390 bp) from 35 Korean isolates showed 99.4% (388/390) similarity with the reference sequence of S. erinaceieuropaei from Korea (G1724; GenBank KJ599680) and an additional 15 Korean isolates revealed 99.2% (387/390) similarity with the reference sequences of S. decipiens from Korea (G1657; GenBank KJ599679). Based on morphologic and molecular databases, the estimated population ratio of S. erinaceieuropaei to S. decipiens was 35: 15. Our results indicate that both S. erinaceieuropaei and S. decipiens found in Korea infect humans, with S. erinaceieuropaei being 2 times more prevalent than S. decipiens. This study is the first to report human sparganosis caused by S. decipiens in humans in Korea.

The life-cycle of Spirometra species from Peninsular Malaysia.

The life-cycle of Malaysian Spirometra spp. was studied under experimental conditions in the laboratory. The Cyclops were reared as the first intermediate host, the hamster as the experimental second intermediate host and cat as the definitive host. Maturation and hatching of eggs took 6 to 12 days by incubation at temperature 30 ºC. The hatched coracidium measured 46 x 34 µm. The Cyclops used were susceptible to the coracidial infection. The procercoid older than 5 days in the Cyclop body cavity had minute spines at the anterior end, calcium corpuscles in the body parenchyma and the cercomer at the posterior end. Procercoids 10 to 14 days old were infective to hamster. The plerocercoids from the hamster after 30 days were long and slender and were infective to cats. The plerocercoids experimentally inoculated to cats developed to adult worms and began to produce eggs between 10 to 60 days. Based on the results that have been obtained, a complete life-cycle was successfully elucidated in the laboratory and hamster was identified to be a good laboratory model for a second intermediate host of Spirometra sp.

Production of polyclonal antibodies against the tegument of sparganum (plerocercoid of Spirometra mansoni) and its immunolocalization.

In a previous study, the author developed a method for separation of the tegument of spargana (plerocercoids of Spirometra mansoni) from the parenchyme using urea. The present study, as a next step, was performed to evaluate which molecules are present in the outer tegument. Two major proteins, 180 and 200 kDa, are present in the tegument and we could make polyclonal antibodies against these molecules. Their immunolocalization was processed and the outermost layer of the spargana showed strong positive staining. Conclusively, we could confirm that the 180 and 200 kDa molecules might be tightly bound membrane proteins in the tegument of spargana.

The Andean hog-nosed skunk Conepatus chinga Molina, 1782 as a new definitive host for Spirometra erinacei Faust, Campbell & Kellog, 1929.

This report describes the finding of Spirometra erinacei Faust, Campbell & Kellog, 1929 (Cestoda, Diphyllobothridae) infecting the small intestine of two Andean hog-nosed skunks (Conepatus chinga Molina, 1782), collected from the locality "Abra La Raya", at Cusco, Peru. Four cestodes were studied and identified as S. erinacei. This is the first report showing that the Andean hog-nosed skunk is one of the natural hosts for this parasite.

Primer hallazgo de Spirometra mansoni en Felis domesticus de Costa Rica / First report of Spirometra mansoni in Felis domesticus from Costa Rica

Se describe un céstodo encontrado en el intestino delgado de gatos provenientes de dos zonas diferentes de Costa Rica (Liberia-Guanacaste y Atenas-Alajuela). De acuerdo con un análisis comparativo morfológico se concluye que los ejemplares hallados corresponden a parásitos del género Spirometra ya que presentan el útero espiralado y huevecillos con los extremos puntiagudos, a diferencia del útero en roseta y huevecillos con extremos redondeados que se encuentran en el género Diphyllobothrium. Dado que presentan un número variable de vueltas en la parte anterior del útero y la vagina es ondulada se clasifican como Spirometra mansoni, siendo éste el primer reporte de este parásito en el país. Se hace una corta revisión de la importancia de este parasito para el ser humano.

[Animal and human cestoda infection (sparganosis)]. / Spirometroz (sparganoz) zhivotnykh i cheloveka.

Spirometrosis (sparganosis) in animals and man is a serious ecological problem in Russia, which requires the constant attention of veterinary and medical services. The paper presents data on the biology of the parasite Spirometra erinacei europaei Muller, 1937, an account and diagram of a developmental cycle, and original photos of parasitic eggs, plerocercoids, strobiles, and cestode joint from animals.

[Strobila periodicity of Spirometra (Cestoda: Diphyllobothriidae)]. / Zur Strobilaperiodizität von Spirometra (Cestoda: Diphyllobothriidae).

The non-induced, endogeneous periodic destrobilation in Spirometra is conceived as an extreme of pseudapolysis. The patent time of S. erinaceieuropaei and S. sp. with single or with simultaneous infestations is arranged in separate patent periods marked off from each other by negative phases. This phenomenon is due to a periodicity of the strobila. In dissections during the negative phase only the scolex and neck alone or provided with a very short strobila could be found in the intestine. There were no striking differences between single and simultaneous infestations. The periodicity altogether, however, showed a great variability. A synchronization of the periodicity of egg shedding and destrobilation is concluded from one simultaneous infestation with demonstrably at least 2 tapeworms. After successive superinvasion, however, the periodicity evidently does not become synchronous, the periodicity of egg shedding is masked.