Analysis of human clinical and environmental Leptospira to elucidate the eco-epidemiology of leptospirosis in Yaeyama, subtropical Japan.

BACKGROUND: Leptospirosis, a zoonosis caused by species in the spirochete genus Leptospira, is endemic to the Yaeyama region in Okinawa, subtropical Japan. Species of the P1 subclade "virulent" group, within the genus Leptospira, are the main etiological agents of leptospirosis in Okinawa. However, their environmental persistence is poorly understood. This study used a combination of bacterial isolation and environmental DNA (eDNA) metabarcoding methods to understand the eco-epidemiology of leptospirosis in this endemic region. FINDINGS: Polymerase chain reaction (PCR) characterized twelve human clinical L. interrogans isolates belonging to the P1 subclade "virulent" subgroup and 11 environmental soil isolates of the P1subclade "low virulent" subgroup (genetically related to L. kmetyi, n = 1; L. alstonii, n = 4; L. barantonii, n = 6) from the Yaeyama region targeting four virulence-related genes (lipL32, ligA, ligB and lpxD1). Clinical isolates were PCR positive for at least three targeted genes, while all environmental isolates were positive only for lipL32. Analysis of infected renal epithelial cells with selected clinical and environmental strains, revealed the disassembly of cell-cell junctions for the Hebdomadis clinical strain serogroup. Comparison of leptospiral eDNA during winter and summer identified operational taxonomic units corresponding to the species isolated from soil samples (L. kmetyi and L. barantonii) and additional P2 subclade species (L. licerasiae, L. wolffii-related, among others) that were not detected by soil cultivation. Total Leptospira read counts were higher in summer than in winter and the analysis of leptospiral/animal eDNA relationship suggested Rattus spp. as a potential reservoir animal. CONCLUSION: Our study demonstrated high environmental Leptospira diversity in the Yaeyama region, particularly during summer, when most of the leptospirosis cases are reported. In addition, several Leptospira species with pathogenic potential were identified that have not yet been reported in Yaeyama; however, the environmental persistence of P1 subclade species previously isolated from human clinical cases in this region was absent, suggesting the need of further methodology development and surveillance.

Insect wing 3D printing.

Insects have acquired various types of wings over their course of evolution and have become the most successful terrestrial animals. Consequently, the essence of their excellent environmental adaptability and locomotive ability should be clarified; a simple and versatile method to artificially reproduce the complex structure and various functions of these innumerable types of wings is necessary. This study presents a simple integral forming method for an insect-wing-type composite structure by 3D printing wing frames directly onto thin films. The artificial venation generation algorithm based on the centroidal Voronoi diagram, which can be observed in the wings of dragonflies, was used to design the complex mechanical properties of artificial wings. Furthermore, we implemented two representative functions found in actual insect wings: folding and coupling. The proposed crease pattern design software developed based on a beetle hindwing enables the 3D printing of foldable wings of any shape. In coupling-type wings, the forewing and hindwing are connected to form a single large wing during flight; these wings can be stored compactly by disconnecting and stacking them like cicada wings.

Topology Optimization-Based Damage Identification Using Visualized Ultrasonic Wave Propagation.

This study proposes a new damage identification method based on topology optimization, combined with visualized ultrasonic wave propagation. Although a moving diagram of traveling waves aids in damage detection, it is difficult to acquire quantitative information about the damage, for which topology optimization is suitable. In this approach, a damage parameter, varying Young's modulus, represents the state of the damage in a finite element model. The feature of ultrasonic wave propagation (e.g., the maximum amplitude map in this study) is inversely reproduced in the model by optimizing the distribution of the damage parameters. The actual state of the damage was successfully estimated with high accuracy in numerical examples. The sensitivity of the objective function, as well as the appropriate penalization exponent for Young's modulus, was discussed. Moreover, the proposed method was applied to experimentally measured wave propagation in an aluminum plate with an artificial crack, and the estimated damage state and the sensitivity of the objective function had the same tendency as the numerical example. These results demonstrate the feasibility of the proposed method.

Freshness estimation of intact frozen fish using fluorescence spectroscopy and chemometrics of excitation-emission matrix.

The current study attempted to provide a convenient, non-invasive and time-saving method to estimate the freshness of intact horse mackerel (Trachurus japonicus) fish in a frozen state using autofluorescence spectroscopy in tandem with multivariate analysis of fluorescence excitation-emission matrices (EEM). The extracted fluorescence data from different freshness conditions were pretreated, masked and reorganized to resolve fish fluorescence spectra from overlapping signals and scattering profiles for detecting and characterizing freshness changes. The real freshness values of the examined fish samples were then traditionally determined by the hard chemical analysis using the high performance liquid chromatography (HPLC) method and expressed as K-values. The fluorescence EEM data and the real freshness values were modeled using partial least square (PLS) regression and a novel algorithm was proposed to identify the ideal combinations of excitation and emission wavelengths being used as perfect predictors. The results revealed that freshness of frozen fish could be accurately predicted with R(2) of 0.89 and root mean square error estimated by cross validation (RMSECV) of 9.66%. This work substantially demonstrated that the autofluorescence spectroscopy associated with the proposed technical approaches has a high potential in non-destructive sensing of fish freshness in the frozen state.