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1.
PLoS Negl Trop Dis ; 14(2): e0008019, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-32101542

RESUMO

BACKGROUND: In China, frogs play an understudied role in the spread of human sparganosis (caused by the larval form of Spirometra). However, our knowledge about the prevalence of sparganum infection in frogs remains fragmented, and the taxonomic identification of the parasite is still controversial. METHODOLOGY/PRINCIPAL FINDINGS: The prevalence of sparganum infection in wild frogs was surveyed at 145 geographical locations from 28 of the 34 provinces/autonomous regions/municipalities in China for six years. The collected sparganum isolates from the different locations were subjected to molecular identification by a multiplex PCR assay and then were analysed with clustering analysis. In the survey, sparganum infection was found in 8 out of 13 of the collected frog species, and the most frequently infected species was Pelophylax nigromaculatus (the infection rate was up to 14.07%). Infected frogs were found in 80 of the 145 surveyed locations. The sparganum infection rates in the wild frogs in several regions of China were still high (above 10%), especially in South and Southwest China. A total of 72 spargana were selected for molecular identification, and the clustering analysis showed that sequences from the Chinese isolates were very similar to those identified as from Spirometra erinaceieuropaei. However, the taxonomy of the genus remains confused and further analysis is required. CONCLUSIONS: Eating wild frogs is associated with considerable health risks in China. Several traditional Chinese folk remedies may increase the risk of infection. The sparganum isolates in China are most likely from S. erinaceieuropaei, but new studies, especially comprehensive morphological analyses, are needed in the future.


Assuntos
Infecções por Cestoides/veterinária , Ranidae/parasitologia , Spirometra/classificação , Animais , Infecções por Cestoides/epidemiologia , Infecções por Cestoides/parasitologia , China/epidemiologia , Interações Hospedeiro-Parasita
2.
Korean J Parasitol ; 57(5): 481-487, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31715688

RESUMO

Mitochondrial DNA sequence variability of Spirometra erinaceieuropaei in GenBank was observed by reinvestigation of mitochondrial cox1 and cytb sequences. The DNA sequences were analyzed in this study, comprising complete DNA sequences of cox1 (n=239) and cytb (n=213) genes. The 10 complete mitochondrial DNA sequences of Spirometra species were compared with those of Korea, China and Japan. The sequences were analyzed for nucleotide composition, conserved sites, variable sites, singleton sites and parsimony-informative sites. Phylogenetic analyses was done using neighbor joining, maximum parsimony, Bayesian inference and maximum-likelihood on cox1 and cytb sequences of Spirometra species. These polymorphic sites identified 148 (cox1) and 83 (cytb) haplotypes within 239 and 213 isolates from 3 Asian countries. Phylogenetic tree topologies were presented high-level confidence values for the 2 major branches of 2 Spirometra species containing S. erinaceieuropaei and S. decipiens, and S. decipiens sub-clades including all sequences registered as S. erinaceieuropaei in cox1 and cytb genes. These results indicated that mitochondrial haplotypes of S. erinaceieuropaei and S. decipiens were found in the 3 Asian countries.


Assuntos
Infecções por Cestoides/parasitologia , DNA de Helmintos/genética , DNA Mitocondrial/genética , Spirometra/genética , Animais , China , Variação Genética , Proteínas de Helminto/genética , Humanos , Japão , Mitocôndrias/genética , Filogenia , Polimorfismo Genético , República da Coreia , Spirometra/classificação , Spirometra/isolamento & purificação
3.
Korean J Parasitol ; 57(5): 513-516, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31715692

RESUMO

Human sparganosis is a food-borne zoonosis mainly caused by the plerocercoid belonging to the genus Spirometra. The most common clinical sign of sparganosis is a subcutaneous mass in the trunk including abdominal or chest wall. The mass may be mistaken for a malignant tumor, thereby causing difficulty in terms of diagnosis and treatment. A 66-year-old woman visited our clinic for the removal of a lipoma-like mass. It was movable, hard, and painless. We identified 2 white mass, measuring 0.2×4 cm and 0.2×1 cm. Pathologic findings indicated the white mass was a sparganum. She recalled having eaten a raw frog approximately 60 years before. A 35-year-old who lived North Korea was also presented to our clinic with an asymptomatic nodule on her abdomen. Intraoperatively, we found sparganum approximately 24 cm size. Subcutaneous masses are associated with clinical signs of inflammation or they may mimic a soft tissue neoplasm. While the incidence rate of sparganosis has decreased with economic development and advancements in sanitation, surgeons still encounter patients with sparganosis in the clinical setting. Therefore, a careful history is required in order to diagnose sparganosis.


Assuntos
Abdome/cirurgia , Lipoma/parasitologia , Esparganose/parasitologia , Spirometra/isolamento & purificação , Adulto , Idoso , Animais , Feminino , Humanos , Lipoma/cirurgia , Esparganose/cirurgia , Spirometra/classificação , Spirometra/genética
4.
Korean J Parasitol ; 57(3): 309-312, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31284356

RESUMO

Spargana were collected from human and frogs in Liaoning and Hubei Provinces, China. PCR amplification and direct sequencing of A cox1 fragment was PCR-amplified from genomic DNA extracted from 7 specimens (5 from humans and 2 from frogs). The cox1 fragment (390 bp) showed 97-100% similarity to the reference sequence of S. erinaceieuropaei and 88-89% to the reference sequence of S. decipiens. There were 1-12 bases different between these worms, but no obvious genetic variation (0-3.3%) to the references. There was little difference of cox1 gene between sparganum samples of humans and frogs (1-3%). This study is the first report on S. erinaceieuropaei spargana from humans in Liaoning and Hubei Provinces.


Assuntos
Anuros/parasitologia , Infecções por Cestoides/parasitologia , Spirometra/genética , Spirometra/isolamento & purificação , Animais , China , Ciclo-Oxigenase 1/genética , Proteínas de Helminto/genética , Humanos , Filogenia , Reação em Cadeia da Polimerase , Spirometra/classificação
5.
Vet Parasitol Reg Stud Reports ; 16: 100270, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-31027603

RESUMO

The goal of this study was to assess the prevalence of gastrointestinal and respiratory parasites of shelter cats from northeast Georgia, thus promoting a more targeted approach in parasite diagnosis and treatment. Fecal samples of cats kept in a shelter located in Lavonia, northeastern Georgia, USA, were processed for the presence of parasites using double centrifugation sugar flotation (n = 103) and Baermann techniques (n = 98). Flotation revealed eggs of Toxocara cati (17.5%), Ancylostoma sp. (11.7%), Taeniidae (3.9%), Spirometra mansonoides (2.9%), Mesocestoides sp. (1%), Dipylidium caninum (1%), and Eucoleus aerophilus (1%), and oocysts of Cystoisospora felis (16.5%), and Cystoisospora rivolta (8.7%). Baermann diagnosed Aelurostrongylus abstrusus larvae in 5 cats (5.1%), while fecal flotation alone identified only 2 of these infections. Taeniidae eggs were identified to species-level by PCR and sequencing targeting the cytochrome oxidase c subunit 1 (cox1) of the mitochondrial DNA. All isolates belong to Hydatigera taeniaeformis sensu stricto, which is the first unequivocal report of the species in North America. Overall, 45.6% of the cats were infected with at least one parasite. This prevalence of infection is much higher than what is generally reported in client owned animals, highlighting the importance of using appropriate fecal diagnostic techniques to detect gastrointestinal and respiratory parasites on newly adopted cats. Correct diagnosis may direct appropriate treatment and control strategies, which would mitigate the risk of infection of other animals in household, and human exposure to zoonotic parasites.


Assuntos
Doenças do Gato/epidemiologia , Gastroenteropatias/veterinária , Doenças Parasitárias em Animais/epidemiologia , Doenças Respiratórias/veterinária , Distribuição por Idade , Ancylostoma/classificação , Ancylostoma/isolamento & purificação , Animais , Doenças do Gato/parasitologia , Gatos , Cestoides/classificação , Cestoides/isolamento & purificação , Fezes/parasitologia , Feminino , Gastroenteropatias/epidemiologia , Gastroenteropatias/parasitologia , Georgia/epidemiologia , Isospora/classificação , Isospora/isolamento & purificação , Funções Verossimilhança , Pneumopatias Parasitárias/epidemiologia , Pneumopatias Parasitárias/parasitologia , Pneumopatias Parasitárias/veterinária , Masculino , Mesocestoides/classificação , Mesocestoides/isolamento & purificação , Doenças Parasitárias em Animais/parasitologia , Filogenia , Prevalência , Doenças Respiratórias/epidemiologia , Doenças Respiratórias/parasitologia , Distribuição por Sexo , Spirometra/classificação , Spirometra/isolamento & purificação , Toxocara/classificação , Toxocara/isolamento & purificação
6.
Korean J Parasitol ; 57(6): 639-645, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31914517

RESUMO

In the present study, a Spirometra species of Tanzania origin obtained from an African leopard (Panthera pardus) and spotted hyena (Crocuta crocuta) was identified based on molecular analysis of cytochrome c oxidase I (cox1) and NADH dehydrogenase subunit I (nad1) as well as by morphological observations of an adult tapeworm. One strobila and several segments of a Spirometra species were obtained from the intestine of an African male leopard (Panthera pardus) and spotted hyena (Crocuta Crocuta) in the Maswa Game Reserve of Tanzania. The morphological characteristics of S. theileri observed comprised 3 uterine loops on one side and 4 on the other side of the mid-line, a uterine pore situated posterior to the vagina and alternating irregularly either to the right or left of the latter, and vesicular seminis that were much smaller than other Spirometra species. Sequence differences in the cox1 and nad1 genes between S. theileri (Tanzania origin) and S. erinaceieuropaei were 10.1% (cox1) and 12.0% (nad1), while those of S. decipiens and S. ranarum were 9.6%, 9.8% (cox1) and 13.0%, 12.6% (nad1), respectively. The morphological features of the Tanzania-origin Spirometra specimens coincided with those of S. theileri, and the molecular data was also consistent with that of S. theileri, thereby demonstrating the distribution of S. theileri in Tanzania. This places the leopard (Panthera pardus) and spotted hyena (Crocuta Crocuta) as new definitive hosts of this spirometrid tapeworm.


Assuntos
Infecções por Cestoides/veterinária , Hyaenidae/parasitologia , Panthera/parasitologia , Spirometra/isolamento & purificação , Animais , Infecções por Cestoides/parasitologia , DNA Mitocondrial/genética , Masculino , Filogenia , Spirometra/classificação , Spirometra/genética , Tanzânia
7.
Infect Genet Evol ; 66: 256-261, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30315893

RESUMO

The incidence of sparganosis, a parasitic disease caused by plerocercoid larvae of the genus Spirometra, has gradually risen worldwide (especially in remote areas) in recent years. Pulmonary and pleural sparganosis, as well as other sites of infestation, including the subcutaneous tissues, the abdominal viscera, brain and eyes, has been reported. In clinical practice, due to the atypical signs and symptoms as well as limited laboratory approaches for the specific detection of sparganum, sparganosis is often misdiagnosed. In the present study, an 11-year-old girl visited the Department of Infectious Diseases in Shanghai Children's Medical Center for recurrent shoulder and chest pain and shortness of breath. Imaging tests demonstrated bilateral pleural and pericardial effusion, enlarged lymph nodes in front of the tracheal carina, and infection of the left lower lobe. Sparganum were not observed in the dissected soft tissue at the root of the right thigh with naked-eye and light microscopy examination. Histologic examination revealed granulomatous inflammation and tunnel-like necrosis with eosinophilic, neutrophilic and lymphocytic infiltration. Although the patient's serum was positive for sparganum antibodies, the diagnosis of sparganosis was not confirmed for more than three months. Ultimately, genomic DNA of Spirometra erinaceieuropaei was detected in the mass at the root of the right thigh using next-generation sequencing (NGS), confirming the diagnosis of sparganosis. The patient was treated with praziquantel (150 mg/kg/day) without recurrence after an eight-month follow-up. We present, for the first time, a study of human sparganosis diagnosed using NGS, which provided a clinically actionable diagnosis of a specific infectious disease from an uncommon pathogen.


Assuntos
Esparganose/diagnóstico , Esparganose/parasitologia , Spirometra/classificação , Spirometra/genética , Animais , Criança , Ecocardiografia , Feminino , Genes de Helmintos , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Derrame Pericárdico , Radiografia Torácica , Análise de Sequência de DNA
8.
Korean J Parasitol ; 56(4): 359-364, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30196668

RESUMO

The taxonomy of Spirometra species has been controversial despite the medical and veterinary importance. Currently, only a few Spirometra species are considered valid species in the genus Spirometra. In the present study, the distribution of Spirometra species obtained from animals in Korea were identified by molecular analysis of the mitochondrial cytochrome c oxidase I (cox1) gene. A total of 28 Spirometra species specimens were analyzed. These were all collected between 1973 and 2008 in the Republic of Korea. Mitochondrial cox1 sequences were examined for a total of 28 specimens comprising 14 S. decipiens and 14 S. ranarum. The difference in partial cox1 sequences (316 bp) between S. erinaceieuropaei (KJ599680) and S. ranarum (this study) was 9.3%, while that between S. decipiens (KJ599679) and S. ranarum (this study) was 2.2%. Genetic analyses identified 2 Spirometra species in animals such as cat, leopard cat, dog, duck and snake in Korea as S. decipiens and S. ranarum. S. decipiens and S. ranarum were present in Gyeongnam Province (P), Jeonnam P, Gangwon P, Chungbuk P, and Seoul. S. decipiens was found in tadpoles, snakes, ducks, cats, leopard cats and dogs, while S. ranarum was found in cats and dogs. The ratio of S. decipiens:S. ranarum calculated from the molecular data was 14:14 (or 1:1). These results indicate that S. decipiens and S. ranarum are sympatrically distributed in Korea.


Assuntos
Infecções por Cestoides/veterinária , Complexo IV da Cadeia de Transporte de Elétrons/genética , Spirometra/classificação , Spirometra/genética , Animais , Sequência de Bases/genética , Gatos , Código de Barras de DNA Taxonômico , DNA Mitocondrial/genética , Cães , Patos/parasitologia , Larva/parasitologia , Mitocôndrias/enzimologia , República da Coreia , Serpentes/parasitologia , Spirometra/isolamento & purificação , Fatores de Tempo
9.
Mol Phylogenet Evol ; 117: 75-82, 2017 12.
Artigo em Inglês | MEDLINE | ID: mdl-28606444

RESUMO

The larva of Spirometra erinaceieuropaei can parasitize humans, causing a serious parasitic zoonosis known as sparganosis. Although it is medically important, our knowledge about the phylogenetic position of S. erinaceieuropaei and its evolutionary history is fragmentary. In this study, complete mitochondrial (mt) genomes of 4 geographically distinct isolates of S. erinaceieuropaei spargana collected from 4 frog hosts (Hylarana guentheri, Rana nigromaculata, R. rugulosa, R. temporaria) were characterized using an Illumina sequencing platform. In addition, all available mt genomes of Cestoda in GenBank were included to reconstruct the phylogeny and to explore the evolutionary history of these tapeworms. The genome features of S. erinaceieuropaei contained 12 protein-coding genes (PCGs), 22 transfer RNA genes, 2 ribosomal RNA genes and 2 non-coding regions. Nucleotide sequences of mtDNA from different frog hosts were similar. Three genes, cox1, cytb and nad4, had high levels of nucleotide diversity. Phylogenetic analyses supported the sibling relationship between Bothriocephalidae and Diphyllobothriidae. Molecular dating analysis indicated that the divergence between Diphyllobothrium and Diplogonoporus started in the late Miocene. The mt genomes of S. erinaceieuropaei will serve as a useful dataset for studying the genetics and systematics of the species of Spirometra genus in particular and diphyllobothriid tapeworms in general.


Assuntos
Genoma Mitocondrial/genética , Genômica , Filogenia , Spirometra/classificação , Spirometra/genética , Animais , DNA Mitocondrial/genética , Genética Populacional , Humanos , Anotação de Sequência Molecular , Ranidae/parasitologia
10.
Emerg Infect Dis ; 23(4): 665-668, 2017 04.
Artigo em Inglês | MEDLINE | ID: mdl-28322697

RESUMO

Human sparganosis is a foodborne zoonosis endemic in Asia. We report a series of 9 histologically confirmed human sparganosis cases in Hong Kong, China. All parasites were retrospectively identified as Spirometra erinaceieuropaei. Skin and soft tissue swelling was the most common symptom, followed by central nervous system lesions.


Assuntos
Esparganose/epidemiologia , Esparganose/parasitologia , Spirometra/isolamento & purificação , Adulto , Idoso , Animais , Feminino , Parasitologia de Alimentos , Hong Kong/epidemiologia , Humanos , Masculino , Pessoa de Meia-Idade , Spirometra/classificação , Spirometra/genética , Zoonoses
11.
Exp Parasitol ; 175: 1-7, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28185895

RESUMO

Although medically important, the systematics of Spirometra and the taxonomic position of S. erinaceieuropaei remain unclear. In this study, the 18S rDNA gene of S. erinaceieuropaei sparganum from naturally infected frogs caught in 14 geographical locations of China was sequenced. In addition, all available 18S sequences of the family Diphyllobothriidae in the Genbank database were included to reconstruct the phylogeny of diphyllobothriid tapeworms. The secondary structure model of the 18S rDNA was also predicated to further explore the sequence variation. Phylogenetic analyses were performed using maximum parsimony (MP), maximum likelihood (ML) and Bayesian inference (BI) methods. The intraspecific divergences of 18S rDNA in Chinese sparganum isolates ranged from 0.0 to 0.4%. Regions of V2, V4 and V7 were the most variable regions in the secondary structure of 18S rDNA. With the exception of genera Duthiersia and Probothriocephalus, other genera (i.e., Adenocephalus, Diphyllobothrium, Diplogonoporus, Duthiersia, Schistocephalus and Spirometra) selected in the Diphyllobothriidae shared similar topologies of V2, V4 and V7 structures. The topology of generated phylogenetic trees revealed close relationships among Adenocephalus, Digramma, Diphyllobothrium, Diplogonoporus, Ligula, Sparganum and Spirometra. The exact phylogenetic position of Spirometra species should be further analyzed with more sampling and more useful molecular markers.


Assuntos
DNA Ribossômico/química , Filogenia , RNA Ribossômico 18S/genética , Spirometra/classificação , Animais , Teorema de Bayes , Cestoides/classificação , Cestoides/genética , China , Funções Verossimilhança , Conformação de Ácido Nucleico , Ranidae , Alinhamento de Sequência , Análise de Sequência de DNA , Plerocercoide/classificação , Plerocercoide/genética , Spirometra/genética
12.
Vet Parasitol Reg Stud Reports ; 10: 58-61, 2017 12.
Artigo em Inglês | MEDLINE | ID: mdl-31014600

RESUMO

The zoonotic helminth, Spirometra, has several species with almost global distribution. Herein, we describe the first detailed molecular detection of Spirometra erinaceieuropaei in a road-killed wildcat (Felis silvestris) in Iran and its identification at the species level using CO1 gene. Genomic DNA was extracted using CTAB extraction method. The DNA then was applied for PCR amplification of cytochrome c oxidase subunit I (CO1) gene. Afterwards, PCR product was sequenced and obtained data were analyzed and multiple aligned using BLAST program, ClustalX and Bioedit software. Microscopy findings and diagnostic clues revealed that the parasite is a Spirometra sp. cestode. Consequently, molecular analysis on the basis of cytochrome c oxidase subunit 1 (CO1) gene demonstrated that the species is Spirometra erinaceieuropaei. Regarding optimum climate conditions and previous reports of animal infection in this region, the likelihood of human involvement should be potentially considered.


Assuntos
Infecções por Cestoides/veterinária , Felis/parasitologia , Spirometra/isolamento & purificação , Animais , Infecções por Cestoides/epidemiologia , Infecções por Cestoides/parasitologia , Irã (Geográfico)/epidemiologia , Spirometra/classificação
13.
Korean J Parasitol ; 54(4): 503-7, 2016 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-27658603

RESUMO

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.


Assuntos
Complexo IV da Cadeia de Transporte de Elétrons/genética , NADH Desidrogenase/genética , Filogenia , Spirometra/classificação , Spirometra/isolamento & purificação , Estruturas Animais/anatomia & histologia , Animais , Análise por Conglomerados , Microscopia , Análise de Sequência de DNA , Homologia de Sequência , Spirometra/anatomia & histologia , Spirometra/enzimologia , Estados Unidos
14.
Acta Trop ; 164: 150-164, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27613585

RESUMO

Spargana are plerocercoid larvae of cestode tapeworms of the genus Spirometra, Family Diphyllobothriidae, parasitic to frogs, reptiles, birds and mammals. This parasitic disease in humans can be transmitted through the use and consumption of amphibians and reptiles. The available knowledge about Spirometra in South America is scarce, and there are only a few reports on the occurrence of sparganum in amphibians and reptiles, many of them published in old papers not easily available to researchers. In this work we present a review on this topic, provide new records in two species of amphibians and 7 species of reptiles from Brazil and Uruguay respectively. We also summarize current knowledge of Spirometra in the continent, along with an updated of host taxonomy. We could gather from the literature a total of 15 studies about amphibian and reptile hosts, published between 1850 and 2016, corresponding to 43 case reports, mostly from Brazil (29) and Uruguay (8), Argentina (3), Peru (2), and Venezuela (1); the majority of them related to reptiles (five lizards and 26 snake species), and 14 corresponded to amphibians (9 anurans). Plerocercoid larvae were located in different organs of the hosts, such as subcutaneous tissue, coelomic cavity, peritoneum, and musculature. The importance of amphibians and reptiles in the transmission of the disease to humans in South America is discussed. Relevant issues to be studied in the near future are the taxonomic characterization of Spirometra in the region and the biological risk of reptile meat for aboriginal and other rural communities.


Assuntos
Anfíbios/parasitologia , Infecções por Cestoides/epidemiologia , Répteis/parasitologia , Spirometra/isolamento & purificação , Animais , Brasil , Infecções por Cestoides/transmissão , Vetores de Doenças , Humanos , América do Sul/epidemiologia , Spirometra/classificação , Uruguai
15.
Korean J Parasitol ; 54(2): 181-5, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-27180576

RESUMO

Human sparganosis is a zoonotic disease caused by infection with larval forms (procercoid/plerocercoid) of Spirometra spp. The purpose of this study was to identify Spirometra spp. of infected snakes using a multiplex PCR assay and phylogenetic analysis of mitochondrial DNA sequence data from the spargana of terrestrial snakes obtained from Korea and China. A total of 283 snakes were obtained that included 4 species of Colubridae comprising Rhabdophis tigrinus tigrinus (n=150), Dinodon rufozonatum rufozonatum (n=64), Elaphe davidi (n=2), and Elaphe schrenkii (n=7), and 1 species of Viperidae, Agkistrodon saxatilis (n=60). The snakes were collected from the provinces of Chungbuk, Chungnam, and Gyeongbuk in Korea (n=161), and from China (n=122). The overall infection rate with spargana was 83% (235/283). The highest was recorded for D. rufozonatum rufozonatum (100%), followed by A. saxatilis (85%) and R. tigrinus tigrinus (80%), with a negative result for E. davidi (0%) and E. schrenkii (0%). The sequence identities between the spargana from snakes (n=50) and Spirometra erinaceieuropaei (KJ599680) or S. decipiens (KJ599679) control specimens were 90.8% and 99.2%, respectively. Pairwise genetic distances between spargana (n=50) and S. decipiens ranged from 0.0080 to 0.0107, while those between spargana and S. erinaceieuropaei ranged from 0.1070 to 0.1096. In this study, all of the 904 spargana analyzed were identified as S. decipiens either by a multiplex PCR assay (n=854) or mitochondrial cox1 sequence analysis (n=50).


Assuntos
Infecções por Cestoides/veterinária , Colubridae/parasitologia , Ciclo-Oxigenase 1/genética , DNA Mitocondrial/genética , Spirometra/classificação , Spirometra/genética , Viperidae/parasitologia , Animais , Sequência de Bases , Infecções por Cestoides/parasitologia , China , DNA de Helmintos/genética , Reação em Cadeia da Polimerase Multiplex , Filogenia , República da Coreia , Análise de Sequência de DNA , Spirometra/isolamento & purificação
16.
Parasitol Int ; 65(5 Pt A): 428-31, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-27235572

RESUMO

Species of the genus Spirometra are diphyllobothriid tapeworms with complex life cycles and are involved in human sparganosis, a neglected disease that affects individuals worldwide. Although some species were reported in wild felids and human cases of sparganosis were described in Brazil, the biology and taxonomy of these parasites are poorly understood. In the present study, samples of diphyllobothriids (eggs and/or proglottids) obtained from the stools of wild carnivores (Leopardus pardalis and Lycalopex vetulus) and plerocercoid larvae found in a snake (Crotalus durissus) from Brazil were analysed by amplifying a fragment of the gene cytochrome c oxidase subunit 1 (cox 1). The DNA sequences obtained here for the first time from the Spirometra spp. from Brazil were used to evaluate the phylogenetic relationships with other species. Molecular data identified two species in the Brazilian samples (evolutionary divergence of 17.8-19.2%). The species were identified as Spirometra sp. 1, found in Le. pardalis, and Spirometra sp. 2 found in Ly. vetulus and C. durissus, and they differed from Asian isolates of Spirometra erinaceieuropaei (17.5-20.2% and 12.2-15.6%, respectively), a species previously considered to be distributed worldwide. Moreover, Spirometra sp. 1 is genetically distinct from Sparganum proliferum from Venezuela (19.6-20.4%), while Spirometra sp. 2 is more closely related with the Venezuelan species (6.1-7.0%). Sequences of Spirometra sp. 2 revealed that it is conspecific with the Argentinean isolate of Spirometra found in Lycalopex gymnocercus (1.9-2.2%). Taxonomic and phylogenetic aspects related to New World species of Spirometra are briefly discussed.


Assuntos
Infecções por Cestoides/veterinária , Crotalus/parasitologia , Felidae/parasitologia , Spirometra/classificação , Spirometra/genética , Animais , Animais Selvagens/parasitologia , Sequência de Bases , Brasil , Infecções por Cestoides/parasitologia , DNA de Helmintos/genética , Complexo IV da Cadeia de Transporte de Elétrons/genética , Tipagem Molecular , Filogenia , Análise de Sequência de DNA
17.
Korean J Parasitol ; 53(4): 455-63, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-26323844

RESUMO

The present study was performed to compare the mitochondrial genomes between 2 Spirometra tapeworms, Spirometra erinaceieuropaei and Spirometra decipiens (Cestoidea: Diphyllobothriidae), which larval stages are important etiological agents of sparganosis in humans. For each species, the full mitochondrial genome was amplified in 8 overlapping fragments using total genomic DNA purified from a single worm as the template. The mitochondrial genomes were 13,643 bp (S. erinaceieuropaei) and 13,641 bp (S. decipiens) in length and contained 36 genes; 12 protein-coding genes, 2 ribosomal RNA (rRNA, small and large subunits), and 22 transfer RNAs (tRNAs). The 12 protein-coding genes constituted 10,083 bp (S. erinaceieuropaei) and 10,086 bp (S. decipiens) of their respective mitochondrial genomes. The tRNA genes, ranging in length from 56 to 70 bp, were identified based on putative secondary structures such as the typical cloverleaf shape. A total of 23 intergenic sequences, varying from 1 to 204 bp in size, were interspersed in S. erinaceieuropaei (total, 504 bp) and S. decipiens (total, 496 bp) mtDNA. The 12 protein-coding genes of S. erinaceieuropaei and S. decipiens differed by 12.4%, whereas the overall difference in mtDNA sequence between S. erinaceieuropaei and S. decipiens was 12.9%. Thus, from the standpoint of the mitochondrial genome, S. decipiens represents a valid species that can be distinguished from S. erinaceieuropaei.


Assuntos
Genoma Helmíntico , Genoma Mitocondrial , Spirometra/genética , Animais , Sequência de Bases , Infecções por Cestoides/parasitologia , DNA Mitocondrial/química , DNA Mitocondrial/genética , Humanos , Dados de Sequência Molecular , Conformação de Ácido Nucleico , Fases de Leitura Aberta , Filogenia , Spirometra/química , Spirometra/classificação
18.
Infect Genet Evol ; 35: 1-8, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26205687

RESUMO

Spirometra erinaceieuropaei is an important diphyllobothriid cestode whose larval plerocercoid (sparganum) can parasitise humans, causing sparganosis. In China, sparganosis is an important foodborne parasitic zoonosis and poses a serious threat to human health. However, our knowledge of the taxonomic classification of S. erinaceieuropaei and its relationship with other diphyllobothriid cestodes, such as Diphyllobothrium, remains fragmentary. In this study, we determined the complete sequences of cytochrome b (cytb) and cytochrome c oxidase subunit I (cox1) genes from 61 isolates of S. erinaceieuropaei collected from 11 geographical locations in eastern China and compared them to the sequences of Diphyllobothrium species from the GenBank database. Then, we evaluated the genetic variation of S. erinaceieuropaei and the phylogenetic relationship between S. erinaceieuropaei and Diphyllobothrium. Both negative Tajima's D and Fu's FS values of neutrality tests and unimodal curve of mismatch distribution analyses supported demographic population expansion. Bayesian skyline plot (BSP) analyses indicated that a population expansion occurred in the Pleistocene. Phylogenetic analysis based on cytb, cox1, and cytb+cox1 under all three methods tested (neighbour-joining, maximum parsimony, and maximum likelihood) suggested that the S. erinaceieuropaei isolates from eastern China and the genus Diphyllobothrium are separate monophyletic groups. Furthermore, the uncorrelated log-normal relaxed molecular-clock model suggested that divergence between Spirometra and Diphyllobothrium began in the late Miocene.


Assuntos
Ciclo-Oxigenase 1/genética , Citocromos b/genética , Diphyllobothrium/classificação , Spirometra/classificação , Animais , Anuros/parasitologia , Teorema de Bayes , China , Diphyllobothrium/genética , Diphyllobothrium/isolamento & purificação , Variação Genética , Proteínas de Helminto/genética , Humanos , Filogenia , Spirometra/genética , Spirometra/isolamento & purificação
19.
Korean J Parasitol ; 53(3): 299-305, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-26174823

RESUMO

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.


Assuntos
Esparganose/parasitologia , Spirometra/genética , Spirometra/isolamento & purificação , Adulto , Idoso , Animais , Doenças do Gato/parasitologia , Gatos , Doenças do Cão/parasitologia , Cães , Complexo IV da Cadeia de Transporte de Elétrons/genética , Feminino , Proteínas de Helminto/genética , Humanos , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Filogenia , República da Coreia/epidemiologia , Esparganose/diagnóstico , Spirometra/anatomia & histologia , Spirometra/classificação , Adulto Jovem
20.
PLoS One ; 10(3): e0119295, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25793277

RESUMO

BACKGROUND: Sparganosis caused by invasion of the plerocercoid larvae (spargana) of Spirometra erinaceieuropaei have increased in recent years in China. However, the population genetic structure regarding this parasite is still unclear. In this study, we used the sequences of two mitochondrial genes cytochrome b (cytb) and cytochrome c oxidase subunit I (cox1) to analyze genetic variation and phylogeographic structure of the S. erinaceieuropaei populations. METHODOLOGY/PRINCIPAL FINDINGS: A total of 88 S. erinaceieuropaei isolates were collected from naturally infected frogs in 14 geographical locations of China. The complete cytb and cox1 genes of each sample was amplified and sequenced. Total 61 haplotypes were found in these 88 concatenated sequences. Each sampled population and the total population have high haplotype diversity (Hd), accompanied by very low nucleotide diversity (Pi). Phylogenetic analyses of haplotypes revealed two distinct clades (HeN+HuN+GZ-AS clade and GX+HN+GZ-GY clade) corresponding two sub-networks yielded by the median-joining network. Pairwise FST values supported great genetic differentiation between S. erinaceieuropaei populations. Both negative Fu's FS value of neutrality tests and unimodal curve of mismatch distribution analyses supported demographic population expansion in the HeN+HuN+GZ-AS clade. The BEAST analysis showed that the divergence time between the two clades took place in the early Pleistocene (1.16 Myr), and by Bayesian skyline plot (BSP) an expansion occurred after about 0.3 Myr ago. CONCLUSIONS: S. erinaceieuropaei from central and southern China has significant phylogeographic structure, and climatic oscillations during glacial periods in the Quaternary may affect the demography and diversification of this species.


Assuntos
Spirometra/classificação , Spirometra/genética , Animais , China , DNA Mitocondrial/genética , Variação Genética , Geografia , Filogenia , Filogeografia
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