Host range differences between two species of freshwater horsehair worm (Nematomorpha: Chordiidae) Chordodes japonensis and C. formosanus in Japan.

Horsehair worms (Nematomorpha: Gordioidea) are endoparasites of terrestrial arthropods, of which two species, Chordodes japonensis and C. formosanus, may have been historically confused due to their morphological similarity. In this study, we conducted field surveys and laboratory parasite inoculation experiments to clarify the characteristics and differences in host range and specificity between the two species. The field surveys revealed that the host ranges of the two species are markedly different. C. formosanus parasitized diverse mantids, while C. japonensis infected only species in the genus Tenodera, as predicted in previous studies. However, the two species had one mantid host, T. angustipennis in common. Parasite inoculation experiments using three species of mantids, revealed parasitism in each of the host-parasite combinations same as in the field study. These results suggest that the observed differences in host patterns are due to physiological factors between the host and parasite. Furthermore, cross-testing conducted on the shared host T. angustipennis showed that the two species coexisted within the same host in some cases, suggesting that the competitive exclusivity of the two species is low. This study also presents a tentative list of host ranges for the two species, comparing and integrating information from this and previous studies. However, the possibility remains that the host ranges of both species may be more diverse.

Host selection and potential predation in the host-parasite interaction between the isopod Tachaea chinensis and freshwater host species.

Tachaea chinensis is an ectoparasite commonly found on diverse ecologically and commercially valuable freshwater shrimps and prawns. Previous studies on this parasite have focused on its distribution and taxonomical identification, while its host preference and/or the potential predation in this host-parasite interaction remained poorly understood. In this study, we investigate the host preference and potential predation of the isopod T. chinensis using manipulative choice and predation experiments under laboratory settings. The preference toward a broad range of host decapods in single-host treatments, indicates a low host specificity, which ultimately aids in the survival of this parasite in the natural environment. Tachaea chinensis responded well to the shrimp Palaemon paucidens when presented with uncommon host species in all three treatments. In host-parasite predation treatments, all the tested P. paucidens shrimp, the prawn Macrobrachium nipponense, and the crayfish Procambarus clarkii were able to consume the isopod-especially the invasive crayfish P. clarkii, which consumed a greater percentage in a considerably shorter time frame (Fisher's exact test, P < 0.01). This study demonstrated for the first time the ability of larger freshwater decapods to prey upon T. chinensis. Despite the large difference in the maximum attainable size of those freshwater species, a high predation pressure by the invasive crayfish on the isopod is anticipated, if they are present in the same environment.

Host Size Selection in the Ectoparasite Tachaea chinensis (Isopoda: Corallanidae) under Laboratory Conditions.

Tachaea chinensis is a temporary ectoparasite infesting freshwater shrimps and prawns in eastern Asia. This study investigated the host size selection by T. chinensis across common freshwater shrimps under laboratory conditions. A total of 70 isopods were allowed to select between host shrimps of different size and different species in pairwise selection experiments. In treatments involving different sizes of the same host species, T. chinensis tended to select the medium host option in all of the four treatments. Similarly, T. chinensis demonstrated greater preference towards medium host (90%) when provided with mixed host options (medium Palaemon paucidens vs small Neocaridina spp.). However, despite the increase in the infestation proportion on medium Neocaridina spp., the isopod significantly selected the small P. paucidens when provided with a choice between medium Neocaridina spp. and small P. paucidens. In manca stage (1 day after hatch) T. chinensis treatment, the isopods showed no specific preference between large and medium Neocaridina spp. These results suggest that T. chinensis is likely to show size specificity according to the developmental stage, a size specificity that ultimately ensures adequate space for isopods' growth while maintaining a minimum risk of predation.

Size Relationship of the Ectoparasite Tachaea chinensis in the Host Shrimp Palaemon paucidens From Shiga Prefecture, Japan.

Monthly sampling of freshwater shrimps infested by the corallanid ectoparasite Tachaea chinensis was conducted at an agricultural canal of the Etsura River system, Shiga Prefecture, Japan, from April 2018 through December 2019. A total of 944 Palaemon paucidens and 279 Neocaridina spp. were collected, and 203 immature T. chinensis were found attached to the carapace of these hosts. Mean prevalence was 23.1% for P. paucidens and 6.4% for Neocaridina spp. Newly hatched T. chinensis ectoparasitizing P. paucidens were detected in August 2018 (minimum body length of parasite 3.2 mm) and appeared continually until June 2019 (maximum body length of 10.3 mm) and were not observed in July. In contrast, T. chinensis on Neocaridina spp. were first detected in August of each year (with minimum body length of 2.0 mm) and were observed until September in 2018 and to November in 2019 (maximum body length of 5.8 mm). Ratios of the body length of T. chinensis to the carapace length of the host shrimp were almost constant (0.8-0.9) with the host species and parasite growth. Although infestation by early-life-stage individuals of T. chinensis was observed on both shrimp species, P. paucidens appeared to be the principal host for individuals larger than 6 mm in body length.

Molecular phylogeny of Japanese marine Tanytarsini chironomids (Chironomidae: Chironominae).

Tanytarsini is a large tribe of Chironomidae with at least 11 recorded marine species grouped in three genera. In this study, we performed a phylogenic analysis using molecular data from 13 Tanytarsini genera, including all Japanese marine species in a large tribe of Chironominae, to estimate their phylogenetic positions and evolutionary history. The phylogenetic reconstruction grouped the marine species in two clades. One clade was composed of five marine Tanytarsus and two marine Pontomyia species within a larger clade of Tanytarsus. Pontomyia is considered to have morphologically specialized and adapted to marine environments by rapid evolution, although it formed a clade with Tanytarsus. The only one species of Tanytarsus, T. pelagicus, clustered as a member of the second clade, which was mainly composed by species of the genus Paratanytarsus. Thus, we redescribe T. pelagicus as Paratanytarsus pelagicus.

16S rRNA Gene Sequence Identification of Cultivable-Bacterioplankton Between Ambient Water and Gastrointestinal Tract (GIT) of Resident Teleost.

This study was conducted to evaluate the ambient water bacterial dynamics and its influence on the gut of resident teleosts; Oreochromis niloticus. The bacterial communities in the ambient water and the gastrointestinal tract (GIT) of O. niloticus were profiled using a culture-dependent method and followed by the 16S rRNA gene sequencing. The results indicated bacterial phyla of Proteobacteria, Actinobacteria, Bacteroidetes, and Firmicutes respectively between the two microbial consortia. However, the relative abundance among the bacterial phyla notably differed between the two consortia. The relative abundance of Proteobacteria (≤ 67%) was dominant in both consortia, but contrarily, Actinobacteria and Bacteroidetes were higher in the ambient water relative to the GIT which indicated Proteobacteria and Bacilli as the highest diversity. Nevertheless, the relative abundance of γ-proteobacteria and Bacilli remarkably dominated both consortia at the class level, with Bacillus and Pseudomonas being the most abundant operational taxonomic units (OTUs).

Antibacterial activity of Mallotus japonicus (L.F.) Müller Argoviensis on growth of Aeromonas hydrophila, A. salmonicida, Edwardsiella tarda and Vibrio anguillarum.

AIMS: The present study evaluated the antimicrobial activities of the medicinal plant Mallotus japonicus against the fish pathogenic bacteria, Aeromonas hydrophila, Aeromonas salmonicida, Edwardisella tarda and Vibrio anguillarum, and also describes the antimicrobial activities of the major and minor active compounds present within the plant extract. The synergistic effects by way of combination of these compounds were also evaluated and described. Chemical constituents of the plant extracts were analysed using the liquid chromatography-mass spectrometry (LC-MS) and described. METHODS AND RESULTS: The diethyl ether-extract of the plant elicited the strongest antibacterial activity against the challenged bacterial species, followed by ethanol- and methanol-extracts. The major active compound of the extracts, bergenin, demonstrated no antibacterial activity, but other compounds in the extracts did. CONCLUSION: Mallotus japonicus could be used as a prophylaxis to treat bacterial disease infections of fishes and its diethyl ether-extract has the potential of an alternative to antibiotic treatment in aquaculture. SIGNIFICANCE AND IMPACT OF THE STUDY: Mallotus japonicus diethyl ether-extract has the potential of an alternative to antibiotic treatment in aquaculture.

Transcriptome analysis of Chironomus sulfurosus larvae living in acidic environments: Insights into molecular mechanisms for acid tolerance.

Larvae of chironomid Chironomus sulfurosus mainly live in acidic rivers near hot springs, suggesting that they naturally select acidic environments as preferred habitats. Here we showed that C. sulfurosus larvae moved toward acidic areas and stayed alive on agar gels with a pH gradient of H2SO4, and the body fluid pH of the homogenized larvae was near neutral even acclimated under the acidic conditions, indicating mechanisms for acid tolerance. In order to gain insights into this mechanism at the molecular level, de novo RNA-seq analysis was performed on C. sulfurosus larvae. As a result, 1,208 genes were found to be significantly up-regulated in larvae acclimated at pH 2.0 compared to controls at pH 7.0. Among the up-regulated genes, ones encoding cuticle proteins, peritrophic matrix proteins, mucus-forming proteins, F-type ATPase subunits, glutathione S transferases, ß-1,3-D-glucan synthetase, hemoglobin, and cytochrome P450 were identified. This transcriptome analysis in conjunction with behavioral and biochemical assays expands our knowledge of gene expression in C. sulfurosus larvae living in acidic environments, which will provide a basis for further studies to elucidate the molecular mechanisms for acid tolerance employed by organisms in nature.

Population Dynamics of Lancelet Branchiostoma japonicum in the Seto Inland Sea, Japan.

The population dynamics of lancelet Branchiostoma japonicum are reported for six sampling sites in the Seto Inland Sea, Japan, for November 2007-2016. Lancelet growth and life span varied spatially, being faster and shorter, respectively, at Stn 3 (off Marugame, western Bisan-seto) than at other sites; average body length at Stn 3 was 36.1 mm for 2-year-old lancelets, and 38.9-42.1 mm for 4-year-old lancelets at sites 1, 2, 4 and 6. Stepwise multiple regression analysis revealed adult growth rate to be significantly positively related to chlorophyll a concentration, and negatively correlated to lancelet density. Density of newly settled and adult lancelets varied spatially. Chlorophyll a concentration best predicted the density of newly settled lancelets, and sediment particle size best predicted that of adults, indicating that factors affecting lancelet density differ during their life history.

A new species of Tanytarsus van der Wulp, 1874 (Diptera: Chironomidae) and the first Japanese record of Tanytarsus ovatus Johannsen, 1932 with DNA barcodes.

A new species of the genus Tanytarsus van der Wulp, 1874 was described and illustrated based on an adult male. Tanytarsus trichovalis sp. nov. belongs to the eminulus species group, and is closely related to T. tamaundecimus Sasa, 1980, T. okuboi Sasa, 1986, and T. tonebeceus Sasa Tanaka, 2000. However, the species is distinguished from these species by the morphology of the hypopygium. We also present the first record of T. ovatus Johannsen, 1932 in Japan. In addition, DNA barcoding of 5 species including T. trichovalis sp. nov., T. ovatus, and other morphologically related species was undertaken. A new key of the eleven Japanese eminulus species group was compiled.

Determination of the serum metallothionein (MT)1/2 concentration in patients with Wilson's disease and Menkes disease.

We have developed an easy and specific enzyme-linked immunoassay (ELISA) for the simultaneous determination of serum metallothinein-1 (MT-1) and 2 (MT-2) in both humans and experimental animals. A competitive ELISA was established using a specific polyclonal antibody against rat MT-2. The antibody used for this ELISA had exhibited the same cross-reactivity with MT in humans and experimental animals. The NH2 terminal peptide of MT containing acetylated methionine was shown to be the epitope of this antibody. The reactivity of this ELISA system with the liver, kidney and brain in MT1/2 knock-out mice was significantly low, but was normal in an MT-3 knock-out mouse. The lowest detection limit of this ELISA was 0.6ng/ml and the spiked MT-1was fully recovered from the plasma. We investigated the normal range of MT1/2 (25-75%tile) in 200 healthy human serum and found it to be 27-48ng/ml, and this was compared with the serum levels in various liver diseases. The serum MT1/2 levels in chronic hepatitis C (HCV) patients were significantly lower than healthy controls and also other liver diseases. In the chronic hepatitis cases, the MT1/I2 levels increased gradually, followed by the progression of the disease to liver cirrhosis and hepatocellular carcinoma. In particular, we found significantly elevated MT1/2 plasma levels in Wilson's disease patients, levels which were very similar to those in the Long-Evans Cinnamon (LEC) rat (model animal of Wilson's disease). Furthermore, a significantly elevated MT1/2 level was found in patients with Menkes disease, an inborn error of copper metabolism such as Wilson's disease.

Determination of metallothionein-3 by a competitive enzyme-linked immunosorbent assay in experimental animals.

An easy and specific enzyme-linked immunoassay (ELISA) for the determination of metallothinein-3 (MT-3) in experimental animals for the research of heavy metal and chemical toxicity has not been reported yet. Therefore, we have developed a competitive ELISA, using a specific monoclonal antibody raised against human recombinant MT-3 (rMT-3). The epitope mapping of the antibody was conducted using mouse, rat, and human MT-3s and peptide fragments of human MT-3. MT-1/2, MT-3 knock-out (KO) mice and human brain and liver were used for the evaluation of the ELISA. A pretreatment method of the tissue homogenates was also examined. The antibody used for the ELISA had the same cross-reactivity with MT-3 in humans and experimental animals. The human MT- 3 NH(2) terminal peptide (Fr. 1-17) was the demonstrated epitope of this antibody. The reactivity of this ELISA in brain homogenate of MT-3 KO mouse was significantly low compared with the wild type and MT-1/2 KO mice. The lowest detection limit of the ELISA was 10 ng/ml and over 80% of the spiked rMT-3 was recovered in the brain homogenate. The assay linearity was intact with a 5-fold dilution in the brain homogenate. The inter- and intra-assay CV was 6.5%, respectively. An effective pretreatment procedure of the tissue homogenate was also established for this MT-3 ELISA. In conclusion, this competitive ELISA is an easy and specific method for measuring the brain MT-3 level in experimental animals.

[Correlation between required quality assurance equipment and radiation therapy institutional level].

Radiotherapy institutions carry out various types of treatments such as conventional radiation treatment, stereotactic radiotherapy (SRT), intensity-modulated radiation therapy (IMRT) and image-guided radiotherapy (IGRT). According to the radiation treatment type, various kinds of quality assurance (QA) equipment are used to maintain the accurate dose delivery to a patient with accurate positioning. The aim of this report was to propose the QA equipment required at various institutional levels. The institution levels were categorized according to the radiation treatment cases. Institutions in the basic category used conventional radiotherapy techniques employing field sizes larger than 4 cm x 4 cm, or they had no full-time radiation oncologist. In addition to the basic level, five other categories were defined which were based on being able to carry out total body irradiation, three-dimensional conformal radiation therapy/SRT, IMRT, and IGRT. Minimum QA equipment requirements were proposed for each level. This proposal will facilitate the purchase of QA equipment for the installation or refurbishing of a linear accelerator.

Temporal and spatial dynamics of a lower-intertidal lancelet population in the Seto Inland Sea, Japan.

The population dynamics of Branchiostoma japonicum, formerly known as B. belcheri, were Investigated from September 2003 to August 2005 in the intertidal zone at Takehara (Hiroshima Prefecture), Seto Inland Sea, Japan. The intertidal population appeared from spring to autumn and disappeared during winter. A laboratory experiment showed that exposure to temperatures below 1 degree C for 2 hours resulted in severe mortality. This low temperature corresponds to the minimum temperature in sediments in the study area. This result suggested either that the intertidal population collapses in winter because of low temperature, or that the lancelets escape from the intertidal to the subtidal zone. Throughout the research period, no lancelets smaller than 10 mm in body length were found, indicating that no larvae settled in the intertidal zone. The intertidal population is probably maintained by the influx of individuals from the neighboring subtidal population. The mean annual density of the lancelets was greatest (10.6 individuals/m(2)) at station 1 nearest the low water mark, and lowest (0.3 Individuals/m(2)) at station 3 furthest from the low water mark. In summer, the water content of the sediments was remarkably lower at station 3 (20.2%) than at station 1 (25.8%). Another laboratory experiment showed that higher mortality occurred from exposure to sediments with a water content less than 25% for 2 hours, comparable to the water content at station 3, suggesting that the spatial distribution of the lancelets upward in the intertidal is restricted by sediment dryness.

Industrial production of difructose anhydride III (DFA III) from crude inulin extracted from chicory roots using Arthrobacter sp. H65-7 fructosyltransferase.

A practical, economical, and industrial process for the enzymatic production of difructose anhydride III (DFA III) was investigated for crude inulin prepared from chicory roots using Arthrobacter sp. H65-7 fructosyltransferase. A comparable level of DFA III production to that from commercial inulin was obtained using crude inulin, suggesting the feasibility of this production process.

Aggressive carcinoid tumors arising in the duodenum, ileum, and rectum with liver metastasis: a report of three cases.

Three aggressive carcinoid tumors with liver metastasis arising in the duodenum (Case 1, 72-year-old man), ileum (Case 2, 72-year-old man), and rectum (Case 3, 67-year-old woman) were studied. Case 1 initially suffered from hemorrhage from an exophytic large tumor and developed a liver metastasis after operation. Cases 2 and 3 first presented as multiple liver metastases. Case 3 had a small submucosal tumor and showed a relatively high cell proliferation activity. Neoplastic cells expressed gastrin in Case 1, serotonin in Case 2, and did not express several peptides in Case 3.

Osteosarcoma arising in the breast.

We report on a 49-year-old woman with osteosarcoma arising in the breast. She had undergone two consecutive excision biopsies for right breast tumors at ages 40 and 42 years. The tumors were diagnosed as a fibroadenoma and a benign phyllodes tumor, respectively. At age 46 years, she noticed a gradually enlarging mass in the same breast. After 3 years, at age 49 years, total mastectomy was performed. The tumor occupied the entire breast and measured 12x9x8.5 cm. The tumor cells were spindle-shaped and pleomorphic, with large, irregular nuclei and distinct nucleoli. Many tumor cells had characteristics of osteoblastic and chondroblastic elements producing osteoid, osseous, and cartilaginous intracellular substances. Pathologic mitoses and apoptotic cells were frequent. Neoplastic cells had infiltrated the skin. Blood and lymph vessel invasion was present. Tumor cells expressed vimentin, osteopontin, vascular endothelial growth factor, CD10, and alkaline phosphatase, but did not express keratin. Chemotherapy was not effective. The patient died of multiple pulmonary metastases 9 months after mastectomy.

Renewal of genetic composition of a lancelet, Branchiostoma belcheri, in the Seto Inland Sea, Japan.

We studied renewal of genetic composition of a lancelet, Branchiostoma belcheri, at a station in the Seto Inland Sea, Japan, using random amplified polymorphic DNA (RAPD) technique. We detected a total of 49 DNA bands from 27 individuals of 5 seasonal samples. Among these, some were specific to individual or seasonal sample. There were no bands common to all individuals. We calculated the Band Sharing Index (BSI) values for each combination of individuals. BSI was in a wide range of 0.00-0.97 and the average BSI between the individuals of a sample was remarkably different from sample to sample (0.37-0.83). In cluster analysis on the basis of BSI, largely 3 clusters were constructed, which finally connected to a single cluster at as low as about 0.2 of BSI. Besides, many clusters were constructed exclusively by a seasonal sample. These results suggest a large gene pool of lancelets in this area, the remarkable differences in gene pool size among the samples and a seasonal renewal of genetic composition at a habitat.

A case report of plasmapheresis in paraneoplastic cerebellar ataxia associated with anti-Tr antibody.

A 53-year-old-male patient was admitted to the Hiroshima University Hospital in August 2001, with a history of progressive cerebellar ataxia, notable by standing and gait disturbances. Brain computed tomography (CT) and magnetic resonance imaging (MRI) showed no signs of ischemic damage. Two weeks later, left axillary lymphadenopathy developed. The pathological finding was Hodgkin's disease (HD; a classical malignant lymphoma in an early stage). Serum antibodies against Purkinje's cells (anti-Tr antibody) were detected by immunohistochemical study. Thus, we diagnosed this as paraneoplastic cerebellar degeneration (PCD) associated HD. We immediately carried out chemotherapy (ABVD therapy), radiotherapy with 20-30 Gy, steroid pulse and intravenous immunoglobulin (IVIg) without neurologic improvement. The patient was subsequently treated by double filtration plasmapheresis (DFPP), leading to a marked improvement of standing and gait disturbances and to a reduction of autoantibodies, which became undetectable. We conclude that DFPP might be useful in treating patients with cerebellar ataxia, and should be considered as a therapeutic choice for it.

Variations in body size of the lancelet Branchiostoma belcheri at different depths in the Seto Inland Sea: effect of food supply on the growth rate.

Variations in body size of the suspension-feeding lancelet Branchiostoma belcheri were examined from April 2001 to December 2001 at different water depths of 10-80 m. The lancelets were abundantly collected (220 indiv./dredge) at water depth of 10 m. The maximum size, ranging from 36.8 to 50.4 mm BL, decreased with increasing water depth. However, the minimum size of 7.4-7.8 mm BL, which corresponds to settling size, did not different with water depth. From seasonal changes in the length-frequency histograms, five cohorts were observed at every water depth. The lancelets grew to 44 mm BL at water depths of 10-20 m and to 34 mm BL at water depth of 80 m with four years of life span. Developmental stages of the gonads at an age of two years showed that no individuals had undeveloped gonads at water depth of 10 m, but 82% of them did at water depth of 80 m. The analysis of the stepwise multiple regressions of monthly growth rate on environmental variables showed that chlorophyll a was the best explanatory variable and showed a significant positive correlation with the growth of 1-2 years age groups. These results suggested that variations in the body length were mainly fluctuated by phytoplankton supply.