Application of stoichiometric CT number calibration method for dose calculation of tissue heterogeneous volumes in boron neutron capture therapy.

BACKGROUND: Monte Carlo simulation code is commonly used for the dose calculation of boron neutron capture therapy. In the past, dose calculation was performed assuming a homogeneous mass density and elemental composition inside the tissue, regardless of the patient's age or sex. Studies have shown that the mass density varies with patient to patient, particularly for those that have undergone surgery or radiotherapy. A method to convert computed tomography numbers into mass density and elemental weights of tissues has been developed and applied in the dose calculation process using Monte Carlo codes. A recent study has shown the variation in the computed tomography number between different scanners for low- and high-density materials. PURPOSE: The aim of this study is to investigate the effect of the elemental composition inside each calculation voxel on the dose calculation and the application of the stoichiometric CT number calibration method for boron neutron capture therapy planning. METHODS: Monte Carlo simulation package Particle and Heavy Ion Transport code System was used for the dose calculation. Firstly, a homogeneous cubic phantom with the material set to ICRU soft tissue (four component), muscle, fat, and brain was modelled and the NeuCure BNCT system accelerator-based neutron source was used. The central axis depth dose distribution was simulated and compared between the four materials. Secondly, a treatment plan of the brain and the head and neck region was simulated using a dummy patient dataset. Three models were generated; (1) a model where only the fundamental materials were considered (simple model), a model where each voxel was assigned a mass density and elemental weight using (2) the Nakao20 model, and (3) the Schneider00 model. The irradiation conditions were kept the same between the different models (irradiation time and irradiation field size) and the near maximum (D1%) and mean dose to the organs at risk were calculated and compared. RESULTS: A maximum percentage difference of approximately 5% was observed between the different materials for the homogeneous phantom. With the dummy patient plan, a large dose difference in the bone (greater than 12%) and region near the low-density material (mucosal membrane, 7%-11%) was found between the different models. CONCLUSIONS: A stoichiometric CT number calibration method using the newly developed Nakao20 model was applied to BNCT dose calculation. The results indicate the importance of calibrating the CT number to elemental composition for each individual CT scanner for the purpose of BNCT dose calculation along with the consideration of heterogeneity of the material composition inside the defined region of interest.

CT number calibration audit in photon radiation therapy.

BACKGROUND: Inadequate computed tomography (CT) number calibration curves affect dose calculation accuracy. Although CT number calibration curves registered in treatment planning systems (TPSs) should be consistent with human tissues, it is unclear whether adequate CT number calibration is performed because CT number calibration curves have not been assessed for various types of CT number calibration phantoms and TPSs. PURPOSE: The purpose of this study was to investigate CT number calibration curves for mass density (ρ) and relative electron density (ρe ). METHODS: A CT number calibration audit phantom was sent to 24 Japanese photon therapy institutes from the evaluating institute and scanned using their individual clinical CT scan protocols. The CT images of the audit phantom and institute-specific CT number calibration curves were submitted to the evaluating institute for analyzing the calibration curves registered in the TPSs at the participating institutes. The institute-specific CT number calibration curves were created using commercial phantom (Gammex, Gammex Inc., Middleton, WI, USA) or CIRS phantom (Computerized Imaging Reference Systems, Inc., Norfolk, VA, USA)). At the evaluating institute, theoretical CT number calibration curves were created using a stoichiometric CT number calibration method based on the CT image, and the institute-specific CT number calibration curves were compared with the theoretical calibration curve. Differences in ρ and ρe over the multiple points on the curve (Δρm and Δρe,m , respectively) were calculated for each CT number, categorized for each phantom vendor and TPS, and evaluated for three tissue types: lung, soft tissues, and bones. In particular, the CT-ρ calibration curves for Tomotherapy TPSs (ACCURAY, Sunnyvale, CA, USA) were categorized separately from the Gammex CT-ρ calibration curves because the available tissue-equivalent materials (TEMs) were limited by the manufacturer recommendations. In addition, the differences in ρ and ρe for the specific TEMs (ΔρTEM and Δρe,TEM , respectively) were calculated by subtracting the ρ or ρe of the TEMs from the theoretical CT-ρ or CT-ρe calibration curve. RESULTS: The mean ± standard deviation (SD) of Δρm and Δρe,m for the Gammex phantom were -1.1 ± 1.2 g/cm3 and -0.2 ± 1.1, -0.3 ± 0.9 g/cm3 and 0.8 ± 1.3, and -0.9 ± 1.3 g/cm3 and 1.0 ± 1.5 for lung, soft tissues, and bones, respectively. The mean ± SD of Δρm and Δρe,m for the CIRS phantom were 0.3 ± 0.8 g/cm3 and 0.9 ± 0.9, 0.6 ± 0.6 g/cm3 and 1.4 ± 0.8, and 0.2 ± 0.5 g/cm3 and 1.6 ± 0.5 for lung, soft tissues, and bones, respectively. The mean ± SD of Δρm for Tomotherapy TPSs was 2.1 ± 1.4 g/cm3 for soft tissues, which is larger than those for other TPSs. The mean ± SD of Δρe,TEM for the Gammex brain phantom (BRN-SR2) was -1.8 ± 0.4, implying that the tissue equivalency of the BRN-SR2 plug was slightly inferior to that of other plugs. CONCLUSIONS: Latent deviations between human tissues and TEMs were found by comparing the CT number calibration curves of the various institutes.

[Development of a Phantom and Analysis Program for Assessment of Positional Accuracy of Image-guided Radiation Therapy].

PURPOSE: We created a phantom and analysis program for the assessment of IGRT positional accuracy. We verified the accuracy of analysis and the practicality of this evaluation method at several facilities. METHOD: End-to-end test was performed using an in-house phantom, and EPID images were acquired after displacement by an arbitrary amount using a micrometer, with after image registration as the reference. The difference between the center of the target and the irradiated field was calculated using our in-house analysis program and commercial software. The end-to-end test was conducted at three facilities, and the IGRT positional accuracy evaluation was verified. RESULT: The maximum difference between the displacement of the target determined from the EPID image and the arbitrary amount of micrometer displacement was 0.24 mm for the in-house analysis program and 0.30 mm for the commercial software. The maximum difference between the center of the target and the irradiation field on EPID images acquired at the three facilities was 0.97 mm. CONCLUSION: The proposed evaluation method using our in-house phantom and analysis program can be used for the assessment of IGRT positional accuracy.

The presence of Ixodes pavlovskyi and I. pavlovskyi-borne microorganisms in Rishiri Island: an ecological survey.

IMPORTANCE: Understanding the ecology of ticks and tick-borne microorganisms is important to assess the risk of emerging tick-borne diseases. Despite the fact that the Ixodes pavlovskyi tick bites humans, we lack information including population genetics and the reason for the inadequate distribution in Japan. A 5-year survey revealed that Rishiri Island, the main stopover in the East Asian Flyway of wild birds in the northern Sea of Japan, was a refuge of I. pavlovskyi. The I. pavlovskyi included two haplogroups, which were supposed to diverge a long time before the island separated from the continent and Hokkaido mainland. The detection of microorganisms from wildlife revealed that wild birds and rodents play a role in diffusion and settlement, respectively, of not only I. pavlovskyi but also I. pavlovskyi-borne microorganisms including Candidatus Ehrlichia khabarensis and Babesia microti US lineage. Various island-specific factors control I. pavlovskyi dominance and tick-borne pathogen maintenance. The results may enable us to explain how tick-borne infectious microorganisms are transported.

Effect of Target Changes on Target Coverage and Dose to the Normal Brain in Fractionated Stereotactic Radiation Therapy for Metastatic Brain Tumors.

Purpose: We evaluated the dosimetric effect of tumor changes in patients with fractionated brain stereotactic radiation therapy (SRT) on the tumor and normal brain using repeat verification magnetic resonance imaging (MRI) in the middle of the treatment period. Methods and Materials: Fifteen large intracranial metastatic lesions with fractionated SRT were scanned employing standardized planning MRI (MRI-1). Repeat verification MRI (MRI-2) were performed during the middle of the irradiation period. Gross tumor volume (GTV) was defined as the volume of the contrast-enhancing lesion on T1-weighted MRI with gadolinium contrast agent. The doses to the tumor and normal brain were evaluated on the MRI-1 scan. Beam configuration and intensity on the initial volumetric modulated arc therapy plan were used to evaluate the dose to the tumor and the normal brain on MRI-2. We evaluated the effect of D98% (percent dose irradiating 98% of the volume) on the GTV using the plans on the MRI-1 and MRI-2 scans. For the normal brain, the V90%, V80%, and V50% (volume of the normal brain receiving >90%, 80%, and 50% of the prescribed dose, respectively) were investigated. Results: Three (20% of the total) and 4 (26% of the total) tumors exhibited volume shrinkage or enlargement changes of >10%. Five (33% of the total) tumors exhibited volume shrinkage and enlargement changes of <10%. Three tumors (20% of the total) showed no volume changes. D98% of the GTV increased in patients with tumor shrinkage because of dose inhomogeneity and decreased in patients with tumor enlargement, with a coefficient of determination of 0.28. The V90%, V80%, and V50% increase with decreasing tumor volumes and were linearly related to the tumor volume difference with a coefficient of determination values of 0.97, 0.98, and 0.97, respectively. Conclusions: Repeat verification MRI for brain fractionated SRT during the treatment period should be considered to reduce the magnitude of target underdosing or normal brain overdosing.

Interfractional Liver Positional Motion Under Exhaled Breath Holding Based on Cone Beam Computed Tomography.

BACKGROUND/AIM: We measured interfractional liver positional motion in liver stereotactic body radiotherapy (SBRT) with exhaled breath holding (BH) based on kilovoltage (kV) cone-beam computed tomography (CBCT) images. PATIENTS AND METHODS: We collected 528 pre-treatment kV-CBCT images from 132 patients who underwent liver SBRT under exhaled BH using the Abches system, a non-electronic contact-based respiratory monitoring device, and analyzed them to investigate interfractional liver positional motion. Planning computed tomography (CT) scans were obtained using the Abches system when the patients were under exhaled BH. Translational 3-degree-of-freedom (DOF) soft-tissue-based image registration was performed using the kV-CBCT images under exhaled BH after 6-DOF vertebral bone image registration. Interfractional liver positional motions in the left-right (LR), anteroposterior (AP), and craniocaudal (CC) directions were defined based on the differences in the position of the liver relative to the vertebral bones. RESULTS: For all fractions, the absolute mean±standard deviation for the interfractional liver positional motion in the LR, AP, and CC directions was 0.7±1.0 mm, 1.0±1.5 mm, and 2.8±3.1 mm, respectively. The liver interfractional systematic/random positional motions in the LR, AP, and CC directions were 0.9/1.2 mm, 1.4/1.8 mm, and 2.9/3.9 mm, respectively. For all fractions, 100.0%, 98.3%, and 86.9% of the interfractional liver positional motions in the LR, AP, and CC directions, respectively, were less than 5 mm. CONCLUSION: CBCT-guided online correction should be used to correct interfractional liver positions errors present in liver SBRT with exhaled BH.

Candidatus Lariskella arthopodarum endosymbiont is the main factor differentiating the microbiome communities of female and male Borrelia-positive Ixodes persulcatus ticks.

Ixodes persulcatus, a hard-bodied tick species primarily found in Asia and Eastern Europe, is a vector of pathogens to human and livestock hosts. Little research has been done on the microbiome of this species, especially using individual non-pooled samples and comparing different geographical locations. Here, we use 16S rRNA amplicon sequencing to determine the individual microbial composition of 85 Borrelia-positive I. persulcatus from the Japanese islands of Hokkaido and Honshu. The resulting data (164 unique OTUs) were further analyzed to compare the makeup and diversity of the microbiome by sex and location, as well as to determine the presence of human pathogens. We found that, while location had little influence, the diversity of I. persulcatus microbiome was predominantly dependent on sex. Males were seen to have higher microbiome diversity than females, likely due to the high presence of endosymbiotic Candidatus Lariskella arthropodarum within the female microbial communities. Furthermore, high read counts for five genera containing potentially human pathogenic species were detected among both male and female microbiomes: Ehrlichia, Borrelia, Rickettsia, Candidatus Neoehrlichia and Burkholderia and co-infections between different pathogens were frequent. We conclude that the microbiome of I. persulcatus depends mainly on sex and not geographical location and that the major difference between sexes is due to the high abundance of Ca. L. arthropodarum in females. We also stress the importance of this tick species as a vector of potential human pathogens frequently found in co-infections.

Influence of different air CT numbers for IVDT on the dose distribution in TomoTherapy MVCT.

This study aims to evaluate the effect of different air computed tomography (CT) numbers of the image value density table (IVDT) on the retrospective dose calculation of head-and-neck (HN) radiotherapy using TomoTherapy megavoltage CT (MVCT) images. The CT numbers of the inside and outside air and each tissue-equivalent plug of the "Cheese" phantom were obtained from TomoTherapy MVCT. Two IVDTs with different air CT numbers were created and applied to MVCT images of the HN anthropomorphic phantom and recalculated by Planned Adaptive to verify dose distribution. We defined the recalculation dose with MVCT images using both inside and outside air of the IVDT as IVDT MVCT inair ${\mathrm{IVDT}}_{\mathrm{MVCT}}^{\mathrm{inair}}$ and IVDT MVCT outair ${\mathrm{IVDT}}_{\mathrm{MVCT}}^{\mathrm{outair}}$ , respectively. Treatment planning doses calculated on kVCT images were compared with those calculated on MVCT images using two different IVDT tables, namely, IVDT MVCT inair ${\mathrm{IVDT}}_{\mathrm{MVCT}}^{\mathrm{inair}}$ and IVDT MVCT outair ${\mathrm{IVDT}}_{\mathrm{MVCT}}^{\mathrm{outair}}$ . The difference between average MVCT numbers ±1 standard deviation on inside and outside air of the calibration phantom was 65 ± 36 HU. This difference in MVCT number of air exceeded the recommendation lung tolerance for dose calculation error of 2%. The dose differences between the planning target volume (PTV): D98% , D50% , D2% and the organ at risk (OAR): Dmax , Dmean recalculated by IVDT MVCT inair ${\mathrm{IVDT}}_{\mathrm{MVCT}}^{\mathrm{inair}}$ and IVDT MVCT outair ${\mathrm{IVDT}}_{\mathrm{MVCT}}^{\mathrm{outair}}$ using MVCT images were a maximum of 0.7% and 1.2%, respectively. Recalculated doses to the PTV and OAR with MVCT showed that IVDT MVCT outair ${\mathrm{IVDT}}_{\mathrm{MVCT}}^{\mathrm{outair}}$ was 0.5%-0.7% closer to the kVCT treatment planning dose than IVDT MVCT inair ${\mathrm{IVDT}}_{\mathrm{MVCT}}^{\mathrm{inair}}$ . This study showed that IVDT MVCT outair ${\mathrm{IVDT}}_{\mathrm{MVCT}}^{\mathrm{outair}}$ was more accurate than IVDT MVCT inair ${\mathrm{IVDT}}_{\mathrm{MVCT}}^{\mathrm{inair}}$ in recalculating the dose HN cases of MVCT using TomoTherapy.

Out of Asia? Expansion of Eurasian Lyme borreliosis causing genospecies display unique evolutionary trajectories.

Vector-borne pathogens exist in obligate transmission cycles between vector and reservoir host species. Host and vector shifts can lead to geographic expansion of infectious agents and the emergence of new diseases in susceptible individuals. Three bacterial genospecies (Borrelia afzelii, Borrelia bavariensis, and Borrelia garinii) predominantly utilize two distinct tick species as vectors in Asia (Ixodes persulcatus) and Europe (Ixodes ricinus). Through these vectors, the bacteria can infect various vertebrate groups (e.g., rodents, birds) including humans where they cause Lyme borreliosis, the most common vector-borne disease in the Northern hemisphere. Yet, how and in which order the three Borrelia genospecies colonized each continent remains unclear including the evolutionary consequences of this geographic expansion. Here, by reconstructing the evolutionary history of 142 Eurasian isolates, we found evidence that the ancestors of each of the three genospecies probably have an Asian origin. Even so, each genospecies studied displayed a unique substructuring and evolutionary response to the colonization of Europe. The pattern of allele sharing between continents is consistent with the dispersal rate of the respective vertebrate hosts, supporting the concept that adaptation of Borrelia genospecies to the host is important for pathogen dispersal. Our results highlight that Eurasian Lyme borreliosis agents are all capable of geographic expansion with host association influencing their dispersal; further displaying the importance of host and vector association to the geographic expansion of vector-borne pathogens and potentially conditioning their capacity as emergent pathogens.

Invasion of fish parasite Prosorhynchoides ozakii (Trematoda: Bucephalidae) into Lake Kasumigaura and surrounding rivers of eastern Japan.

In 2019 to 2021, the golden mussel Limnoperna fortunei and several freshwater fishes were sampled from 22 sites of the Tone River system including Lake Kasumigaura, Honshu, Japan, to examine the invasion of bucephalid trematodes. The parasite species identification was performed by morphological observation and DNA barcoding based on the sequences of nuclear 28S rDNA and mitochondrial cytochrome c oxidase subunit 1 (cox1). A total of 1719 mussels were collected from 10 sites, and trematode-infected mussels were detected from 8 sites with prevalences between 0.3 and 42.9%. The sporocysts and cercariae were identified as Prosorhynchoides ozakii, a newly introduced species in the river system. A total of 700 fish individuals belonging to 24 species were collected from 15 sites. Two species of catfishes (Silurus asotus and Ictalurus punctatus) harbored mature or immature adults of Pr. ozakii in the intestine with prevalences between 8.3 and 20% including both host species. The metacercariae of Pr. ozakii were found from the fins and epidermis of 13 fish species from 10 sites (prevalence 4.8-100%). Fishes were heavily infected with metacercariae in fins, which were surrounded by the infiltration of hemocytes and rodlet cells. A population genetic analysis of Pr. ozakii did not show an obvious bottleneck, suggesting the possibility that the parasite was intentionally and repeatedly introduced into the river system.

Improved treatment robustness of postoperative breast cancer radiotherapy including supraclavicular nodes.

A combination of a three-dimensional conformal radiation therapy (3D-CRT) plan with a dose gradient of the chest wall area and a volumetric modulated arc therapy (VMAT) plan of the supraclavicular area might improve the dose distribution robustness in the junction. To investigate the impact of patient motion on the dose distribution, hybrid 3D-CRT and VMAT plans were recalculated by shifting the isocenter of the VMAT plan. Compared to the nominal plan, the target D98% for high- vs low-dose gradients decreased by 24% vs 12%. Hybrid VMAT with a low-dose gradient 3D-CRT plan was found to be robust towards patient motion.

Phylogeography of Rhabdias spp. (Nematoda: Rhabdiasidae) collected from Bufo species in Honshu, Shikoku, and Kyushu, Japan including possible cryptic species.

The genus Rhabdias Stiles & Hassall, 1905 comprises lung parasites of amphibians and reptiles worldwide. In Japan, 9 species have been recorded, including Rhabdias incerta Wilkie, 1930 which has been reported only in Bufo species. In this study, to assess the diversity of R. incerta, we performed molecular analyses of Rhabdias species sampled from three species/subspecies of Japanese toads namely Bufo japonicus, B. japonicus formosus, and B. torrenticola, collected in various regions of Honshu, Shikoku, and Kyushu, Japan. DNA sequence divergence was compared using mtDNA (COI) and nuclear DNA (28S) to identify possible cryptic species. Morphological analysis was performed through light microscopy and scanning electron microscopy (SEM). The results revealed that Bufo spp. serve as hosts for at least three Rhabdias species. Morphologically, most samples were identified as R. incerta but with a longer body and larger buccal cavity than originally described. Rhabdias incerta appears to be specific to the genus Bufo and is further subdivided into two or three phylogroups based on subspecies divisions and biogeography of their host. Some Rhabdias specimens collected in this study resemble R. tokyoensis Wilkie, 1930, parasitic in hosts from the order Caudata, which suggests host switching. Both molecular and morphological analyses suggested the presence of undescribed and cryptic Rhabdias species within toads collected in Japan. This study was the first to molecularly characterize Rhabdias species in Japan, including novel sequences of R. incerta and two undescribed species.

Resolution of cryptic species complexes within the genus Metagonimus (Trematoda: Heterophyidae) in Japan, with descriptions of four new species.

A nationwide fish survey was conducted in Japan to detect metacercariae of the genus Metagonimus (Trematoda: Heterophyidae). The metacercariae were subjected to DNA barcoding for molecular species identification. A phylogeny inferred from the sequences of mitochondrial cytochrome c oxidase subunit 1 (cox1) prompted us to recognize three cryptic species complexes (i.e., the M. miyatai complex, the M. takahashii complex, and the M. katsuradai complex). Each complex included one or two undescribed species. For morphological description, adult flukes of each species were raised through the experimental infections of immunosuppressed mice. We propose M. saitoi n. sp., M. kogai n. sp., M. shimazui n. sp., and M. kinoi n. sp., based on their phylogeny, morphology, biogeography, and ecology (host-parasite relationships). The originally described species, M. miyatai, was split into M. miyatai sensu stricto and M. saitoi n. sp. The former is distributed mainly in eastern Japan and uses the sweetfish (Plecoglossus altivelis) and daces (Pseudaspius hakonensis and Ps. sachalinensis) as principal second intermediate hosts, while the latter is in western Japan and its principal fish hosts are the dark chub (Nipponocypris temminckii) and the pale chub (Opsariichthys platypus). The present survey resolves a long-standing controversy on the microtaxonomy of Metagonimus in Japan since the first discovery of Metagonimus yokogawai in 1912, and shows that 10 species of Metagonimus are still distributed in Japan, although human metagonimiasis is almost eradicated.

Stoichiometric CT number calibration using three-parameter fit model for ion therapy.

PURPOSE: Treatment planning for ion therapy involves the conversion of computed tomography number (CTN) into a stopping-power ratio (SPR) relative to water. The purpose of this study was to create a CTN-to-SPR calibration table using a stoichiometric CTN calibration model with a three-parameter fit model for ion therapy, and to demonstrate its effectiveness by comparing it with a conventional stoichiometric CTN calibration model. METHODS: We inserted eight tissue-equivalent materials into a CTN calibration phantom and used six CT scanners at five radiotherapy institutes to scan the phantom. We compared the theoretical CTN-to-SPR calibration tables created using the three-parameter fit and conventional models to the measured CTN-to-SPR calibration table in three tissue types: lung, adipose/muscle, and cartilage/spongy bone. We validated the estimated SPR differences in all cases and in a worst-case scenario, which revealed the largest estimated SPR difference in lung tissue. RESULTS: For all cases, the means ± standard deviations of the estimated SPR difference for the three-parameter fit method model were -0.1 ± 1.0%, 0.3 ± 0.7%, and 2.4 ± 0.6% for the lung, adipose/muscle, and cartilage/spongy bone, respectively. For the worst-case scenario, the estimated SPR differences of the conventional and the three-parameter fit models were 2.9% and -1.4% for the lung tissue, respectively. CONCLUSIONS: The CTN-to-SPR calibration table of the three-parameter fit model was consistent with that of the measurement and decreased the calibration error for low-density tissues, even for the worst-case scenario.

REDISCOVERY OF MICHAJLOVIA TURDI (DIGENEA: BRACHYLAIMOIDEA) FROM JAPAN.

Michajlovia turdi (Yamaguti, 1939) (Digenea: Brachylaimoidea) has been found from the Japanese thrush, Turdus cardis Temminck, 1831, in Hokkaido, the northernmost island of Japan. This is a rediscovery of M. turdi in Japan after approximately 80 years from the original description of the species as Leucochloridium turdiYamaguti, 1939. Here we redescribe the morphology of M. turdi and generate DNA barcodes for the species by sequencing nuclear ribosomal RNA (18S and 28S) and mitochondrial cytochrome c oxidase subunit I (COI) genes. Our molecular phylogenetic analysis was not effective at determining the taxonomic rank of Michajlovia in Brachylaimoidea, and consequently the genus remains as incertae sedis.

PHILOPHTHALMUS HECHINGERI N. SP. (DIGENEA: PHILOPHTHALMIDAE), A HUMAN-INFECTING EYE FLUKE FROM THE ASIAN MUD SNAIL, BATILLARIA ATTRAMENTARIA.

Two cases of human philophthalmosis have been reported in Japan. Gravid flukes removed from the eyes of the patients were broken, but their morphological characteristics suggest that an unknown species of the genus Philophthalmus is involved as a pathogen for humans. The mitochondrial DNA barcode of the human eye fluke enabled us to discover its larval stage from the Japanese mud snail, Batillaria attramentaria. The discovered cercaria had previously been temporarily described as "Philophthalmid sp. I." In this study, we examined the infection status of B. attramentaria with Philophthalmid sp. I found on a muddy seashore of the Seto Inland Sea, Japan, and the resulting metacercariae were experimentally administered to Japanese quails to develop them into the gravid adult stage. The complete specimens of the adult and larval stages allowed us to describe a new species. Based on morphological and molecular analyses, Philophthalmus hechingeri n. sp. is proposed for the human-infecting eye fluke in Japan. The natural definitive hosts of the new species are unknown. However, the habitat of B. attramentaria suggests that shorebirds (seagulls, sandpipers, and plovers) might be the possible candidates.

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.

Treatment planning comparison between dynamic wave arc and volumetric modulated arc therapies for prostate-cancer treatment.

The aim of this study was to compare the quality of dynamic wave arc (DWA) and coplanar volumetric modulated arc therapy (co-VMAT) plans for the treatment of localized prostate cancer. The planning target volume (PTV)-rectum, a section of the PTV comprising the PTV minus that of the rectum, received 78 Gy in 39 fractions as the mean dose to the PTV-rectum. The DWA and co-VMAT plans were generated for each patient using the RayStation treatment planning system for the Vero4DRT system. The PTV-rectum dose (D95%: the percent dose irradiating 95% of the volume), homogeneity index (HI), conformity index (CI), as well as doses to the bladder wall, rectum wall (V10-70 Gy: the percent volume receiving 10-70 Gy), and bilateral femoral heads of the DWA and co-VMAT plans were compared. The output monitor unit (MU) and delivery time obtained for each set of plans were also investigated. In terms of target coverage, the DWA plans provided an average D95% of 75.5 Gy, which was comparable to the co-VMAT-plan D95% of 75.2 Gy (p < 0.05). The HI was significantly better with the DWA. As for the DWA plans, the bladder-wall volume receiving 10, 20, 30, and 40 Gy (V10-40 Gy) was significantly smaller than that of the co-VMAT plans, and the volume of the rectal wall receiving 10 Gy (V10Gy) was significantly larger than that of the co-VMAT plans. The DWA plans yielded a reduced dose to the bilateral femoral heads compared with the co-VMAT plans (p < 0.05). The values of the CI and MU, and the delivery time exhibited no significant differences between the DWA and co-VMAT plans. The DWA plan is a feasible treatment option for prostate cancer radiotherapy.

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.

The phylogeographic puzzle of Pseudoacanthocephalus toshimai, an amphibian acanthocephalan in northern Japan.

The amphibian acanthocephalan, Pseudoacanthocephalus toshimai, was considered to be an island-endemic species in Hokkaido, Japan. However, the parasite was found from Rana ornativentris, Rana tagoi, Zhangixalus arboreus, and Bufo japonicus formosus in northern Honshu (Aomori and Iwate Prefectures), which is separated from Hokkaido by the Tsugaru Strait. The mitochondrial DNA-based phylogenetic and population genetic analyses of P. toshimai showed that the northern Honshu isolates are far distantly related to the Hokkaido isolates, and that a demographic population expansion occurred in Hokkaido during the recent geological past. The rich genetic diversity of P. toshimai in northern Honshu suggests a scenario that anuran hosts invaded Hokkaido together with P. toshimai via the land bridge of the Tsugaru Strait. However, the evolutionary history of Rana pirica, a main definitive host for P. toshimai in Hokkaido, is contradictory to the introduction scenario inferred from the parasite. The finding of several geographically mismatched isolates of P. toshimai from both northern Honshu and Hokkaido suggests a possibility that the migration of the parasite infrequently occurred between the two areas even after the land bridge disappeared. More detailed information on the evolutionary history of anurans is needed to resolve the biogeographical enigma of P. toshimai.