An unknown species of Leucochloridium (Trematoda: Leucochloridiidae) from northern Honshu,Japan

Pulsating broodsacs of Leucochloridium sp. (Trematoda: Leucochloridiidae) were found from amber snails (Succinea lauta) in Iwate, the northern part of Honshu, Japan. A pattern with red-brown vertical stripes was characteristic of the broodsac. Very similar broodsacs were already detected from Okinawa Islands, the southern archipelago of Japan, and tentatively identified as Leucochloridium cf. passeri. A phylogenetic analysis based on DNA sequences of mitochondrial cytochrome c oxidase subunit 1 (cox1) showed that Leucochloridium sp. is different at species level from L. cf. passeri and that both species are related to Leucochloridium vogtianum from Europe. In this study the definitive identification of larval Leucochloridium sp. was impossible, but the resulting phylogeny confirmed that at least 4 species of Leucochloridium are distributed in Japan, depending on locality and climate. The DNA barcode generated in this study will be useful in detecting the adult stage of Leucochloridium sp. from birds.     

Use of ITS rDNA for discrimination of European green- and brown-banded sporocysts within the genus Leucochloridium Carus, 1835 (Digenea: Leucochloriidae)

Transmission of Leucochloridium species to their definitive avian hosts may be facilitated by the rhythmic movement of coloured sporocyst broodsacs in the ocular tentacles of infected snails. These broodsacs resemble caterpillars and by enticement increase the probability of predation by birds. Broodsac banding pattern and colour (green, yellow to red/brown) have traditionally formed part of the taxonomic criteria for the genus. In this study, sequence divergence of the 5.8S rDNA gene and associated internal transcribed spacers (ITS1 and ITS2) in two of the most frequently observed Leucochloridium taxa from Europe is related to broodsac type based on colour and banding pattern. The present green-banded broodsac (L. paradoxum Carus, 1835) and brown-banded broodsac forms (L. variae McIntosh, 1932) differ in ITS sequence by 6.8%, confirming their distinctness. No intraspecific differences were noted within each colour morph in specimens collected from Poland, Denmark or Norway, indicating that a single taxon of each type occurs in Europe. The significance of these findings to our understanding of metapopulation dynamics and evolutionary ecology of Leucochloridium is discussed.     

A revision of the family Leucochloridiidae Poche (Digenea) and studies on the morphology of Leucochloridium paradoxum Carus, 1835

Summary The sporocyst and metacercaria of Leucochloridium paradoxum Carus, recovered from Succinea putris L. in Norway, are described and figured. Adults recovered from experimental infection of Taeniopygia guttata (Vieillot) with L. paradoxum metacercariae are described, using results from scanning electron microscopy, and compared with earlier information. The family Leucochloridiidae Poche is revised. Four genera are recognized within the family (i.e. Leucochloridium Carus, 1835; Urogonimus Monticelli, 1888; Urotocus Looss, 1899; Michajlovia Pojmanska, 1973). The larval ecological and morphological characteristics are tabulated for the three genera with known larval stages. The status of the genus Neoleucochloridium Kagan, 1951, is changed, on the grounds of larval similarities with Leucochloridium Carus. This means that within the genus Leucochloridium Carus three subgenera are erected, based primarily on sporocyst characteristics. These are Leucochloridium, Neoleucochloridium and Papilloleucochloridium. The diagnostic larval and adult characters are tabulated. The known larval records of species in the genus Leucochloridium are discussed and revised within the framework of this new classification. ac]19790812     

First detection of the fruit fly Bactrocera synnephes (Diptera: Tephritidae) on Iriomote Island, south-western Japan

Bactrocera synnephes, a fruit fly species common in Taiwan, was first detected in Japan in 2006, when several male adults were trapped at Kanokawa, an uninhabited coastal area of Iriomote Island, Okinawa (200 km from Taiwan). To examine potential colonization by B. synnephes, we collected fruit of the wild gourd Diplocyclos palmatus (Cucurbitaceae), a host of the fruit fly, as well as other cucurbitaceous species, in Kanokawa and other areas of Iriomote Island. No B. synnephes infestation was found on any cucurbitaceous plants. Based on these results, we conclude that B. synnephes had very recently invaded Iriomote Island, possibly from Taiwan, but had failed to colonize. Bactrocera synnephes may have entered Iriomote Island by natural or human‐induced means, but a human‐induced invasion pathway is less likely in light of the non‐pest status of the species in Taiwan. The disappearance of B. synnephes from Iriomote Island may be related to the loss of host fruit as a result of tropical cyclones. Although B. synnephes is not regarded as a pest elsewhere, the pest status of the species is unclear in Okinawa, where no fruit fly pests currently attack commercial cucurbitaceous fruit. We must remain alert for possible re‐invasion by B. synnephes.     

Brachylaima succini sp. nov. (Trematoda: Brachylaimidae) from Succinea lauta,an amber snail in Hokkaido,Japan

Through a continuous survey of trematodes in land snails of Hokkaido, the northernmost island of Japan, we have discovered four species of the genus Brachylaima (Trematode: Brachylaimidae). Among them, Brachylaima ezohelicis, Brachylaima asakawai, and Brachylaima lignieuhadrae have already been described. Each of the three species is a strict specialist in selecting a particular species of land snail as the first intermediate host. In this report, we propose the fourth species, Brachylaima succini sp. nov., based on ecological, morphological, and phylogenetic considerations. Sporocysts and metacercariae of the new species were found exclusively from Succinea lauta, which is known as an amber snail indigenous to Hokkaido. Phylogenetic trees of nuclear 28S rDNA and mitochondrial cytochrome c oxidase subunit 1 (cox1) demonstrated it to be distinct from the other sympatric species. Although metacercariae of the new species possessed unique morphological characters, adult worms experimentally raised from the metacercariae were similar to those of B. ezohelicis and B. lignieuhadrae. Natural definitive hosts of the new species are unknown, but the existence of common cox1 haplotypes from far-distant localities suggests a possibility that birds are involved as the definitive hosts. Findings of amber snails coinfected with both sporocysts of the new species and Leucochloridium perturbatum also support the involvement of birds.     

Comparison of carbon and nitrogen content of infected and uninfected snails, Succinea ovalis, and the trematode Leucochloridium variae.

In June, 6.7% of adult Succinea ovalis collected near Urbana, Ohio, were infected with the trematode, Leucochloridium variae. The effects of parasitism were assessed as total organic carbon (equivalent to calorific values) and as total nitrogen. The parasite represents 23.8% of total (parasite + snail tissue) dry tissue weight, 21.4% of total carbon and 19.8% of total nitrogen of infected snails. The higher C : N ratio for parasite tissue indicates a higher proportion of nonproteinaceous compounds (e.g., fats and/or carbohydrates) as compared to host tissue. There is less snail tissue in parasited S. ovalis. The C : N ratios for parasitized and nonparasitized snail tissue suggest identical percentage compositions of proteins, fats, and carbohydrates.     

Use of enzyme electrophoresis for differentiating Schistosoma nasale and S. spindale infections of Indoplanorbis exustus in Sri Lanka

Indoplanorbis exustus is the intermediate snail host of both Schistosoma nasale and S. spindale in Sri Lanka. Due to the similar morphology of the cercariae difficulties exist in identifying the species of schistosome infecting snails at transmission sites. Isoelectric focusing in polyacrylamide gels of malate dehydrogenase, glucose phosphate isomerase, phosphoglucomutase and acid phosphatase has been carried out on extracts of cercariae and potentially useful diagnostic markers have been identified. Cellulose acetate membrane electrophoresis has been used to separate two glucose phosphate isomerase phenotypes each of which appears to be diagnostic for one of the species. This simple electrophoretic technique has been used to identify schistosome infections in Indoplanorbis exustus collected at two transmission sites in Sri Lanka. Out of a total of 1,240 snails collected, 13 snails were shown to be infected with S. nasale, 19 with S. spindale and two snails were found to have mixed infections.     

Fasciola hepatica in Snails Collected from Water-Dropwort Fields using PCR

Fasciola hepatica is a trematode that causes zoonosis mainly in cattle and sheep and occasionally in humans. Fascioliasis has been reported in Korea; however, determining F. hepatica infection in snails has not been done recently. Thus, using PCR, we evaluated the prevalence of F. hepatica infection in snails at 4 large water-dropwort fields. Among 349 examined snails, F. hepatica-specific internal transcribed space 1 (ITS-1) and/or ITS-2 markers were detected in 12 snails and confirmed using sequence analysis. Morphologically, 213 of 349 collected snails were dextral shelled, which is the same aperture as the lymnaeid snail, the vectorial host for F. hepatica. Among the 12 F. hepatica-infected snails, 6 were known first intermediate hosts in Korea (Lymnaea viridis and L. ollula) and the remaining 6 (Lymnaea sp.) were potentially a new first intermediate host in Korea. It has been shown that the overall prevalence of the snails contaminated with F. hepatica in water-dropwort fields was 3.4%; however, the prevalence varied among the fields. This is the first study to estimate the prevalence of F. hepatica infection using the vectorial capacity of the snails in Korea.     

Further new species of the plant bug genus Campylomma Reuter from the Oriental Region (Heteroptera: Miridae: Phylinae: Nasocorini)

The fauna of the nasocorine plant bug genus Campylomma Reuter from the Oriental Region, including subtropical Japanese Ryukyu Islands (Okinawa Island of Southwest Islands-group) and Taiwan is updated. Three new species are herein described, C. astica n. sp. (from Okinawa and Taoyuan, Taiwan), C. hibiscicola n. sp. (Bangkok, Thailand) and C. nanrenana n. sp. (Pingtung, Taiwan). Of these, C. astica and C. hibiscicola, were observed to have cryptic habitat-preference, inhabiting bracts, stipules or flower buds of the sea (or coastal) hibiscus, Hibiscus tiliaceus (L.) (Malvaceae), planted for landscaping at urbanized zones. An updated checklist of the 22 Oriental Campylomma species is provided.     

Brachylaima phaedusae n. sp. (Trematoda: Brachylaimidae) from door snails in Japan

The metacercarial infections of door snails (Gastropoda: Clausiliidae) with unknown species of the genus Brachylaima (Trematoda: Brachylaimidae) have recently been reported in eastern Honshu and Kyushu, Japan. A large scale snail survey was carried out to clarify their taxonomic status. From the period of 2015 to 2020, a total of 1239 land snails (768 door snails and 471 others) were collected from 32 localities in Honshu, Shikoku, and Kyushu. The resulting trematode isolates were identified as Brachylaima sp. by mitochondrial DNA barcoding. The sporocysts were found only a few from Megalophaedusa sublunellata (Clausiliidae), Tauphaedusa subaculus (Clausiliidae), and Aegista trochula (Camaenidae), while the metacercariae were frequently detected from 14 species of Clausiliidae and 2 species of other families. Although Brachylaima sp. showed a broad range of intermediate hosts, door snails seem to be very important to drive the life cycle. The gravid adults of Brachylaima sp. was experimentally raised from metacercariae using immunosuppressed mice. Morphological, phylogenetical, and ecological considerations prompted us to propose Brachylaima phaedusae n. sp. for this unknown species. The definitive hosts of the new species are completely unknown. The wide geographic distribution and high genetic diversity of the new species suggest a possibility that the definitive host is ground-foraging birds, which prefer door snails.     

Genome-Wide Scan and Test of Candidate Genes in the Snail Biomphalaria glabrata Reveal New Locus Influencing Resistance to Schistosoma mansoni

BackgroundNew strategies to combat the global scourge of schistosomiasis may be revealed by increased understanding of the mechanisms by which the obligate snail host can resist the schistosome parasite. However, few molecular markers linked to resistance have been identified and characterized in snails.Conclusions/SignificanceThe loci RADres and sod1 both have strong effects on resistance to S. mansoni. Future approaches to control schistosomiasis may benefit from further efforts to characterize and harness this natural genetic variation.     

Notes on some plant-inhabiting fungi collected from the Nansei Islands (2)

In this paper, four fungi collected in Nansei Islands are reported. One is Strasseria garciniae, which must be transferred to the genus Phyllosticta. This fungus was collected in Okinawa Main Island and Iriomote Island. The other three fungi are newly added to the Japanese fungal flora. Phyllosticta ghaesembillae on Codiaeum variegatum and Cercospora asplenii on Asplenium antiquum were collected in Yoron Island. The last one, Coniella australiensis on Eucalyptus robusta was collected in Okinawa Main Island. Their morphology and symptoms on the host plant are described, with some mycological notes.     

Wild host plants of four spider mite species (Acari: Tetranychidae) infesting fruit crops in Okinawa

To determine the wild host plants of four major spider mite species infesting fruit crops grown in the subtropical Asian region, we collected and identified mites from non-crop plants throughout the islands of Okinawa, southwestern Japan. Although the two mango pests, Oligonychus coffeae and O. biharensis, are polyphagous, they did not share any wild host species in the field, indicating that their source plants are completely separate in Okinawa. Several major wild hosts were determined for Eutetranychus africanus, a pest of papaya and citrus. Its host range partially overlapped with that of O. biharensis. The citrus red mite, Panonychus citri, was very rare on non-crop plants in Okinawa, suggesting that it maintains its population mainly on the citrus trees in the area. These results are of great significance when considering vegetation control as part of the integrated management of these pest mites. During our survey, two non-pest species, Panonychus caglei (new to Japan) and Oligonychus gotohi (new to Okinawa), were also found.     

Ecology of bacterial communities in the schistosomiasis vector snailBiomphalaria glabrata

The internal colony-forming bacterial flora of the schistosome intermediate host snailBiomphalaria glabrata (Say) has been characterized in ca. 500 individual snails from Puerto Rico, Guadeloupe, and St. Lucia, and from laboratory aquaria. Freshly captured wild snails harbor 2–40×10⁶ CFU·g^–1, and healthy aquarium snails harbor 4–16×10⁷ CFU·g^–1, whereas moribund individuals have 4–10 times as many bacteria as healthy individuals from the same habitats.Pseudomonas spp. are the most common predominant bacteria in normal snails, whereasAcinetobacter, Aeromonas, andMoraxella spp. predominate in moribund snails. External bacterial populations in water appear to have little effect on the composition and size of the flora in any snail. In addition to normal (healthy) and moribund snails, a third group of snails has been distinguished on the basis of internal bacterial density and predominating genera. These high-density snails may have undergone stresses and may harbor opportunistic pathogens. The microfloras of wild and laboratory-reared snails can be altered and stimulated to increase in density by crowding the snails or treating them with antibiotics.     

Fingerprint of Biomphalaria arabica, the intermediate host of Schistosoma mansoni in Saudi Arabia, using RAPD-PCR

In the time schistosomisis control programs are implemented in many countries, schistosomiasis continues to spread throughout the world. Among these control strategies is the vector control. Within this context, analysis of the genetic variability of the intermediate host snails is important because it allows identification of specific sequences of the genome of this mollusk related to determine their fingerprint. We investigated Biomphalaria arabica, which is found in Saudi Arabia, the intermediate host of Schistosoma mansoni infection. Genetic fingerprint was studied by RAPD-PCR using our own different random primers as well as published primers. The electrophoretic patterns resulting from amplification showed specific polymorphic markers of B. arabica. This information will be helpful in the identification of the snails and demonstrating that RAPD-PCR is an appropriate and efficient methodological approach for establishment of genetic barcode development.     

Shell mats associated with microsporida in Biomphalaria glabrata: genetic studies

Multilayered mats have been observed lining the shell of some Biomphalaria glabrata infected with Steinhausia brachynema. Occurrence of the mats is associated with extensive infection with the microsporidia in these snails, in contrast to most B. glabrata in which infection appears restricted to the intestinal wall. Observations suggest that this variation in pathology may be influenced by a genetic difference in the host snails.     

Detection of Ehrlichia risticii, the Agent of Potomac Horse Fever, in Freshwater Stream Snails (Pleuroceridae: Juga spp.) from Northern California

Ehrlichia DNA was identified by nested PCR in operculate snails (Pleuroceridae: Juga spp.) collected from stream water in a northern California pasture in which Potomac horse fever (PHF) is enzootic. Sequencing of PCR-amplified DNA from a suite of genes (the 16S rRNA, groESL heat shock operon, 51-kDa major antigen genes) indicated that the source organism closely resembled Ehrlichia risticii, the causative agent of PHF. The minimum percentage of Juga spp. harboring the organism in the population studied was 3.5% (2 of 57 snails). No ehrlichia DNA was found in tissues of 123 lymnaeid, physid, and planorbid snails collected at the same site. These data suggest that pleurocerid stream snails may play a role in the life cycle of E. risticii in northern California.     

Studies on the prolonged storage of Metarhizium anisopliae conidia: effect of temperature and relative humidity on conidial viability and virulence against mosquitoes

The shell lengths, dried shell weights, soft tissue wet weights, and soft tissue dry weights were ascertained for noninfected Ilyanassa obsoleta and for the same snail species naturally infected with the following digeneans: Himasthla quissetensis, Zoogonus lasius, Cercaria dipterocerca, Lepocreadium setiferoides, Microphalloides nassicola, Stephanostomum tenue, and Microbilharzia variglandis. Analyses of the data obtained indicate that infection with all of the trematodes listed above causes neither enhanced growth of soft tissues nor accelerated lengthening of the shell. However, snails infected with sporocysts of Z. lasius have significantly heavier shells than do noninfected snails. These data suggest that infection with Z. lasius may have caused enhanced growth in the form of increased calcium deposition in the shell of I. obsoleta, possibly as a result of parasitic castration. In addition, the mean dry weight of the soft tissues of snails infected with M. variglandis, when normalized for shell length, is significantly lower than that of noninfected snails, possibly as a result of parasite-induced pathology. Finally, comparisons among infected snails indicate a trend toward increased soft tissue dry weight in snails infected with L. setiferoides, although the mean dry weight of these snails does not differ significantly from the mean dry weight of noninfected snails.     

Detection of Ancestry Informative HLA Alleles Confirms the Admixed Origins of Japanese Population

The polymorphisms in the human leukocyte antigen (HLA) region are powerful tool for studying human evolutionary processes. We investigated genetic structure of Japanese by using five-locus HLA genotypes (HLA-A, -B, -C, -DRB1, and -DPB1) of 2,005 individuals from 10 regions of Japan. We found a significant level of population substructure in Japanese; particularly the differentiation between Okinawa Island and mainland Japanese. By using a plot of the principal component scores, we identified ancestry informative alleles associated with the underlying population substructure. We examined extent of linkage disequilibrium (LD) between pairs of HLA alleles on the haplotypes that were differentiated among regions. The LDs were strong and weak for pairs of HLA alleles characterized by low and high frequencies in Okinawa Island, respectively. The five-locus haplotypes whose alleles exhibit strong LD were unique to Japanese and South Korean, suggesting that these haplotypes had been recently derived from the Korean Peninsula. The alleles characterized by high frequency in Japanese compared to South Korean formed segmented three-locus haplotype that was commonly found in Aleuts, Eskimos, and North- and Meso-Americans but not observed in Korean and Chinese. The serologically equivalent haplotype was found in Orchid Island in Taiwan, Mongol, Siberia, and Arctic regions. It suggests that early Japanese who existed prior to the migration wave from the Korean Peninsula shared ancestry with northern Asian who moved to the New World via the Bering Strait land bridge. These results may support the admixture model for peopling of Japanese Archipelago.     

Keys to the blow flies of Taiwan,with a checklist of recorded species and the description of a new species of Paradichosia Senior-White (Diptera,Calliphoridae)

Blow flies (Diptera: Calliphoridae) show a great diversity in behavior and ecology, play important roles in ecosystems, and have medical and forensic importance to humans. Despite this, the taxonomy and classification of Taiwan''s Calliphoridae have rarely been studied. In this study, specimens of Taiwanese calliphorids were collected and carefully studied, and all 76 species recorded in Taiwan are listed following the identification keys. Dichotomous keys to all subfamilies, tribes, genera, and species of blow flies recorded in Taiwan are provided, including 16 species that are newly recorded from Taiwan. In addition, one new species of the genus Paradichosia Senior-White is described and illustrated. We also discuss the morphological differences between the specimens of Silbomyia hoeneana Enderlein collected from China and Taiwan, a species that has only been found previously in Southern China.