Identification of species of Diplogonoporus balaenopterae and D. grandis (Cestoda, Diphyllobothriidae)

Comparative study of diplogonadal diphillobothriids from different species of whalebone whales and from man (Japan) and analysis of literary data has made it possible to establish their identity. Only the species Diplogonoporus balaenopterae (L?nnberg, 1892 capable of infecting whalebone whales, dogs and man can be regarded as really existing. The species D. grandis (Blanchard, 1894) Lühe, 1899 is its synonym.     

Complete mitochondrial genomes of Diplogonoporus balaenopterae and Diplogonoporus grandis (Cestoda: Diphyllobothriidae) and clarification of their taxonomic relationships

Although the diplogonadic human tapeworm, Diplogonoporus grandis, has long been considered to be a synonym of the whale tapeworm, Diplogonoporus balaenopterae, the identity of the both species at the complete mitochondrial genomes and nuclear DNA levels has been not sufficiently undertaken to date. In the present study, to clarify the taxonomic relationships between D. balaenopterae and D. grandis at the molecular level, the complete mitochondrial genomes of both species were sequenced and compared. In addition, the genetic variation in the mitochondrial cytochrome c oxidase subunit 1 gene (cox1) and the nuclear internal transcribed spacer-1 (ITS-1) region of the ribosomal RNA gene were examined. The complete mitochondrial genomes of D. balaenopterae and D. grandis consisted of 13,724 bp and 13,725 bp, respectively. These mitochondrial genomes contained 12 protein-coding, 22 transfer RNA and 2 ribosomal RNA genes and two longer non-coding regions. Except for Hymenolepis diminuta, the genomic organization in both species was essentially identical to that in other cestode genomes examined to date. However, differences were observed between Diplogonoporus and Diphyllobothrium species in abbreviated stop codons, sequences and the number of repeat units in the 2nd non-coding regions. The genetic differences observed in the mitochondrial genomes, cox1 and ITS-1 regions of both species were considered typical of intraspecific variation. In conclusion, D. balaenopterae is a taxonomically valid species and D. grandis is a junior synonym of D. balaenopterae based on the zoological nomenclature. Further, molecular-phylogenetic analysis confirmed that D. balaenopterae is more closely related to Diphyllobothrium stemmacephalum, the type-species of the genus Diphyllobothrium, and the taxonomical validity of the genera Diplogonoporus and Diphyllobothrium was also discussed.     

Diphyllobothrium stemmacephalum (Cestoda: Diphyllobothriidae) found from a harbor porpoise in northern Japan,with comments on a geographical gap with human infection cases in southern Japan

Even though the cetacean tapeworm Diphyllobothrium stemmacephalum occurs in both cold and warm waters, human infections and final host occurrences have been confined to temperate areas in and near Japan. We recently obtained a strobila of this cestode that was excreted from a harbor porpoise accidentally caught offshore of Hokkaido of northern Japan. Genetic analysis of 28S rDNA and cox1 genes confirmed that the cestode was D. stemmacephalum. Our finding sets the northernmost record of D. stemmacephalum in the western Pacific, suggesting that the risk of human infections by this parasite in northern Japan deserves further attention.     

Isolation and identification of non-chlorinated phenylbenzotriazole (non-ClPBTA)-type mutagens in the Ho River in Shizuoka Prefecture, Japan

We previously identified 2-[2-(acetylamino)-4-amino-5-methoxyphenyl]-5-amino-7-bromo-4-chloro-2H-benzotriazole (PBTA) congeners as major mutagens in water concentrates from several rivers that flow in three different areas, i.e. Kyoto, Aichi, and Fukui Prefectures, in Japan. In synthesis studies, these PBTAs were shown to be formed from corresponding dinitrophenylazo dyes via non-chlorinated derivatives (non-ClPBTAs). However, only non-ClPBTA-1, i.e. 2-[2-(acetylamino)-4-[bis(2-methoxyethyl)amino]-5-methoxyphenyl]-6-amino-4-bromo-2H-benzotriazole, had been detected as a minor contaminant in the Nishitakase River in Kyoto. In this study, analysis of mutagens in water concentrate from the Ho River, which flows through an area with a textile dyeing industry in Shizuoka Prefecture, Japan, allowed the isolation of four compounds (I, II, III, and IV). These four mutagens were identified as 2-[2-(acetylamino)-4-[N-(2-cyanoethyl)ethylamino]-5-methoxyphenyl]-6-amino-4-bromo-2H-benzotriazole (non-ClPBTA-2), 2-[2-(acetylamino)-4-[(2-hydroxyethyl)amino]-5-methoxyphenyl]-6-amino-4-bromo-2H-benzotriazole (non-ClPBTA-3), 2-(2-acetylamino-4-amino-5-methoxyphenyl)-6-amino-4-bromo-2H-benzotriazole (non-ClPBTA-4), and 2-[2-(acetylamino)-4-(diethylamino)-5-methoxyphenyl]-6-amino-4-bromo-2H-benzotriazole (non-ClPBTA-7) by spectral data and co-chromatography using synthesized standards. Non-ClPBTA-3 and -7 were highly mutagenic in Salmonella typhimurium YG1024, inducing 159,000 and 178,000 revertants/microg, respectively, in the presence of S9 mix. Like PBTAs, non-ClPBTAs might have been produced from azo dyes during industrial processes in dyeing factories and released into rivers.     

Characterization of the complete mitochondrial genome sequence of Spirometra erinaceieuropaei (Cestoda: Diphyllobothriidae) from China

Sparganosis, caused by the plerocercoid larvae of members of the genus Spirometra, can cause significant public health problem and considerable economic losses. In the present study, the complete mitochondrial DNA (mtDNA) sequence of Spirometra erinaceieuropaei from China was determined, characterized and compared with that of S. erinaceieuropaei from Japan. The gene arrangement in the mt genome sequences of S. erinaceieuropaei from China and Japan is identical. The identity of the mt genomes was 99.1% between S. erinaceieuropaei from China and Japan, and the complete mtDNA sequence of S. erinaceieuropaei from China is slightly shorter (2 bp) than that from Japan. Phylogenetic analysis of S. erinaceieuropaei with other representative cestodes using two different computational algorithms [Bayesian inference (BI) and maximum likelihood (ML)] based on concatenated amino acid sequences of 12 protein-coding genes, revealed that S. erinaceieuropaei is closely related to Diphyllobothrium spp., supporting classification based on morphological features. The present study determined the complete mtDNA sequences of S. erinaceieuropaei from China that provides novel genetic markers for studying the population genetics and molecular epidemiology of S. erinaceieuropaei in humans and animals.     

Mass Death of Predatory Carp,Chanodichthys erythropterus,Induced by Plerocercoid Larvae of Ligula intestinalis (Cestoda: Diphyllobothriidae)

We describe here the mass death of predatory carp, Chanodichthys erythropterus, in Korea induced by plerocercoid larvae of Ligula intestinalis as a result of host manipulation. The carcasses of fish with ligulid larvae were first found in the river-edge areas of Chilgok-bo in Nakdong-gang (River), Korea at early February 2016. This ecological phenomena also occurred in the adjacent areas of 3 dams of Nakdong-gang, i.e., Gangjeong-bo, Dalseong-bo, and Hapcheon-Changnyeong-bo. Total 1,173 fish carcasses were collected from the 4 regions. To examine the cause of death, we captured 10 wondering carp in the river-edge areas of Hapcheon-Changnyeong-bo with a landing net. They were 24.0-28.5 cm in length and 147-257 g in weight, and had 2-11 plerocercoid larvae in the abdominal cavity. Their digestive organs were slender and empty, and reproductive organs were not observed at all. The plerocercoid larvae occupied almost all spaces of the abdominal cavity under the air bladders. The proportion of larvae per fish was 14.6-32.1% of body weight. The larvae were ivory-white, 21.5-63.0 cm long, and 6.0-13.8 g in weight. We suggest that the preference for the river-edge in infected fish during winter is a modified behavioral response by host manipulation of the tapeworm larvae. The life cycle of this tapeworm seems to be successfully continued as the infected fish can be easily eaten by avian definitive hosts.     

Morphological observation and molecular phylogeny of Spirometra decipiens complex 1 (Cestoda: Diphyllobothriidae) found in cat from Chile

The systematics of tapeworms in the genus Spirometra has been progressing with the accumulation of molecular genetics data, but the taxonomic status of many nominal species remains under debate. We report morphological and molecular-phylogenetic data for a Spirometra species collected from a domestic cat (Felis silvestris catus) in Chiloé Island, Chile. The Spirometra species was shown to be genetically conspecific with Spirometra decipiens complex 1 found in a Pampas fox (Lycalopex gymnocercus) from Argentina, and was closely related to a Hoary fox (Lycalopex vetulus) and rattlesnake (Crotalus durissus) from Brazil. Therefore, the presence of S. decipiens complex 1 was molecularly confirmed for the first time in Chile. The findings of the present study add useful information for the systematics of poorly known Spirometra species in South America.     

Seroepidemiological study of norovirus infection in Aichi Prefecture, Japan

The serological prevalence of IgG antibody to seven NoV strains (GI.1, GI.4, GII.3, GII.4, GII.10, GII.12 and GII.15) among inhabitants aged 1–62 years of Aichi Prefecture, Japan was studied. Age-related seroprevalence was measured by ELISA using baculovirus-expressed recombinant VLP antigens. Seropositive rates for all seven VLP antigens gradually increased with age. Among the tested antigens, the highest seropositive rate was for the GII.4 strain. This result is consistent with the recent epidemic of NoV infection due to GII.4 strain in Japan.     

Rapid development of resistance to diamide insecticides in the smaller tea tortrix,Adoxophyes honmai (Lepidoptera: Tortricidae), in the tea fields of Shizuoka Prefecture,Japan

We investigated the susceptibility of the smaller tea tortrix, Adoxophyes honmai Yasuda, to diamide insecticides in the Shimada-Yui tea fields in Shizuoka Prefecture, Japan, from 2006 to 2011. By 2011, the insects had developed significant resistance even to concentrations far above the registration concentrations of two diamides, flubendiamide and chlorantraniliprole. The lethal concentration 50 (LC₅₀) values of flubendiamide showed a rapid annual increase from 16.2 ppm in 2007 to 161 ppm in August 2011, exceeding the registration concentration of 100 ppm in 2010 and 2011. The LC₅₀ values of chlorantraniliprole increased sharply from 25.3 ppm in 2010 to 98.8 ppm in August 2011, exceeding the registration concentration of 50 ppm. The LC₅₀ values for flubendiamide and chlorantraniliprole at 10 days after treatment in insects collected in August 2011 were 105-fold and 77.2-fold higher, respectively, than those in a susceptible strain.     

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 ({"type":"entrez-nucleotide","attrs":{"text":"MK955901","term_id":"1796114604","term_text":"MK955901"}}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.     

Seasonal occurrence of Laodelphax striatellus (Hemiptera: Delphacidae) in a rice-forage crops mixed cropping area in central Kyushu,Japan

The small brown planthopper, Laodelphax striatellus (Fallén) is an important pest of rice in East Asia because it is a vector of rice stripe virus. In this study, we examined the seasonal occurrence of L. striatellus in an agricultural area in central Kyushu in which cropping systems have been changing since the 1970s. The first generation of L. striatellus emerged in the spring, not only on barley, oats, and Italian ryegrass but also on spring crops of forage maize. The second generation of L. striatellus adults emerged in forage maize fields in July, after harvesting of winter crops of barley, oats, and Italian ryegrass. These results suggest that spring crops of forage maize are important hosts of L. striatellus from spring to early summer. The overwintering generation of L. striatellus was collected from rice ratoon, suggesting this is a favorable host for L. striatellus during winter. The appearance of these novel hosts may be a factor in the recent re-emergence of L. striatellus in central Kyushu.     

Geography and host specificity: two forces behind the genetic structure of the freshwater fish parasite Ligula intestinalis (Cestoda: Diphyllobothriidae)

Parasite species with global distributions and complex life cycles offer a rare opportunity to study alternative mechanisms of speciation and evolution in a single model. Here, genealogy and genetic structure, with respect to geography and fish host preference, have been analyzed for Ligula intestinalis, a tapeworm affecting freshwater fish. The data analyzed consisted of 109 tapeworms sampled from 13 fish host species in 18 different localities on a macrogeographic scale. Two mitochondrial genes, cytochrome oxidase subunit I and cytochrome B, and the nuclear sequence of intergenic transcribed spacer 2 (ITS2) were used for the genetic reconstruction. Different evolutionary patterns were found at the local and at the global geographic scales. On a local scale, the flat genetic structure was mainly attributed to contiguous range expansion. Migrating birds are the most likely cause of the homogenisation of the whole population, preventing the creation of significant genetic barriers. By contrast, on a global scale, genetically distant and well-separated clusters are present in different geographic areas. Reproductive isolation was found even between clades living in sympatry and infecting the same definitive host, suggesting the existence of efficient biologically determined genetic barriers, and thus possibly separate species. Although the ITS2 sequences were found to display considerable intragenomic variability, their relationships were generally in good agreement with the topology derived from mitochondrial genes.     

Two Taenia species found in Japan, with new distribution record of Taenia polyacantha Leuckart, 1856 (Cestoda: Taeniidae)

In an epidemiological survey for Echinococcus multilocularis in rodents and insectivores from the northernmost part of the central mainland of Japan (Honshu), two taeniid species, Taenia crassiceps and Taenia polyacantha, were found in Microtus montebelli and Apodemus argenteus, respectively. The latter is the first record of distribution in Japan, and the former is the second after its first recovery from the central part of Japan. Although we have found neither larval nor strobilar stage of E. multilocularis there, discovery of these taeniid species, having overlapping global distribution with E. multilocularis in red foxes, Vulpes vulpes as well as multiple occurrences of hydatid patients having no history of visits to the endemic areas shows the possibility that the life-cycle of E. multilocularis might be maintained at least in the northernmost part of Honshu.     

Genetic characterization of Dibothriocephalus latus and Dibothriocephalus dendriticus (Cestoda: Diphyllobothriidae) from Chile based on haplotype analysis using mitochondrial DNA markers

Dibothriocephalus latus and Dibothriocephalus dendriticus are found throughout the temperate and sub-arctic zones of the northern hemisphere, but they are also found in the southern core countries of South America, Chile and Argentina. Genetic characteristics of D. latus and D. dendriticus from South America have yet to be fully defined. The present study aimed to understand the genetic characteristics of D. latus and D. dendriticus from Chile by haplotype network analysis of mitochondrial cytochrome c oxidase subunit I gene (cox1) and cytochrome b gene (cob), as well as their origins. Dibothriocephalus latus and D. dendriticus plerocercoid larvae were obtained from feral and/or wild salmonids captured in Lake Llanquihue in Región de Los Lagos, and Lake Panguipulli in Región de Los Ríos, located south of central Chile. Haplotype analysis of D. latus revealed that H1 in cox1 and H2 in cob are the key haplotypes common to D. latus across the world, including Chile, and both genes exhibited limited genetic diversity in D. latus. It was assumed that D. latus was brought into South America by European and Russian immigrants in the 19th century as previously reported. In contrast, both the cox1 and cob of D. dendriticus display considerable genetic diversity, with no common haplotypes between D. dendriticus populations from Chile and the northern hemisphere. More intriguingly, two cob haplotypes (H24, H25) detected in Chilean D. dendriticus were closely linked to haplotypes (H30, H31) detected in North American D. dendriticus, strongly implying that D. dendriticus in Chile was brought by piscivorous migrating birds from North America. It has also been estimated that the D. dendriticus from Chile genetically diverged from the D. dendriticus from the northern hemisphere approximately 1.11 million years ago, long before humans migrated to the southern parts of South America.     

Prevalence of human immunodeficiency virus (HIV) infection among hemophiliacs in Fukuoka, Japan

To determine when the hemophiliacs in Fukuoka prefecture, Japan, first became positive for antibodies, we tested human immunodeficiency virus (HIV) antibodies on serum samples obtained from 1976-1987 stored at -30 C. Fifteen out of 64 hemophilia A patients (23.4%), five out of 11 hemophilia B patients (45.5%), but none of 17 patients with von Willebrand's disease (0%) were positive for HIV antibodies. In this series, two with hemophilia A became positive for HIV antibodies for the first time in 1983, and in 1984 another four with hemophilia A and one with hemophilia B became positive.