Investigating Immunotoxicity in Black Carp (Mylopharyngodon piceus) Fingerlings Exposed to Niclosamide.

Niclosamide (NIC) is a potent salicylanilide molluscicide/helminthicide commonly utilized for parasite and mollusc control in aquatic environments. Due to its persistent presence in water bodies, there is growing concern regarding its impact on aquatic organisms, yet this remains inadequately elucidated. Consequently, this study aims to assess the hepatotoxic effects and detoxification capacity of black carp (Mylopharyngodon piceus) in a semi-static system, employing various parameters for analysis. NIC was applied to juvenile black carp at three different concentrations (0, 10 and 50 µg/L) for 28 days in an environmentally realistic manner. Exposure to 50 µg/L NIC resulted in an increase in hepatic lysozyme (LYZ), alkaline phosphatase (ALP), and complement 4 (C4) levels while simultaneously causing a decrease in peroxidase (POD) activity. Additionally, NIC exposure exhibited a dose-dependent effect on elevating serum levels of LYZ, ALP, complement 3 (C3), C4, and immunoglobulin T (IgT). Notably, the mRNA levels of immune-related genes tnfα, il8, and il6, as well as nramp and leap2, were upregulated in fish exposed to NIC. RNA-Seq analysis identified 219 differentially expressed genes (DEGs) in M. piceus after NIC exposure, with 94 upregulated and 125 downregulated genes. KEGG and GO analyses showed enrichment in drug metabolism pathways and activities related to oxidoreductase, lip oprotein particles, and cholesterol transport at 50 µg/L NIC. Additionally, numerous genes associated with lipid metabolism, oxidative stress, and innate immunity were upregulated in NIC-exposed M. piceus. Taken together, these findings indicate that NIC has the potential to cause hepatotoxicity and immunotoxicity in M. piceus. This research offers important insights for further understanding the impact of molluscicide/helminthicide aquatic toxicity in ecosystems.

Niclosamide subacute exposure alters the immune response and microbiota of the gill and gut in black carp larvae, Mylopharyngodon piceus.

Niclosamide (NIC) is a commonly used insecticide and molluscicide in the prevention and treatment of parasitic diseases in fish. The utilization of NIC has the potential to disrupt the microbial community present on the mucosal tissue of fish, leading to localized inflammatory responses. The objective of this study was to evaluate the impact of NIC on the immune system and bacterial populations within the gill and gut of Mylopharyngodon piceus. Fish were subjected to varying concentrations of NIC, including a control group (0 µg/L), a low NIC group (15% 96 h LC50, LNG, 9.8 µg/L), and a high NIC group (80% 96 h LC50, HNG, 52.5 µg/L). Gill and gut samples were collected 28 days post-exposure for analysis. The findings revealed that the 96-h LC50 for NIC was determined to be 65.7 µg/L, and histopathological examination demonstrated that exposure to NIC resulted in gill filament subepithelial edema, exfoliation, degeneration, and a decrease in gill filament length. Furthermore, the gut exhibited apical enterocyte degeneration and leucocyte infiltration following NIC exposure. Additionally, NIC exposure led to a significant elevation in the levels of immunoglobulin M (IgM), complement component 3 (C3), and complement component 4 (C4) in both gill and gut tissues. Moreover, the activity of lysozyme (LYZ) was enhanced in the gill, while the activities of peroxidase (POD) and immunoglobulin T (IgT) were increased in gut tissue. The exposure to NIC resulted in enhanced mRNA expression of c3, c9, tnfα, il6, il8, and il11 in the gill tissue, while decreasing c3 and il8 expression in the gut tissue. Furthermore, the natural resistance-associated macrophage protein (nramp) mRNA increased, and liver-expressed antimicrobial peptide 2 (leap2) mRNA decreased in gill and gut tissues. And hepcidin (hepc) mRNA levels rose in gill but fell in gut tissue. NIC exposure also led to a decrease in gill bacterial richness and diversity, which significantly differed from the control group, although this separation was not significant in the gut tissue. In conclusion, the administration of NIC resulted in alterations in both the immune response and mucosal microbiota of fish. Furthermore, it was noted that gills displayed a heightened vulnerability to sublethal effects of NIC in comparison to gut tissues.

The Effect of Short-Term Artificial Feed Domestication on the Expression of Oxidative-Stress-Related Genes and Antioxidant Capacity in the Liver and Gill Tissues of Mandarin Fish (Siniperca chuatsi).

To investigate whether Mandarin fish developed oxidative stress after being domesticated with artificial feed, we conducted a series of experiments. Oxidative stress is an important factor leading to diseases and aging in the body. The liver integrates functions such as digestion, metabolism, detoxification, coagulation, and immune regulation, while the gills are important respiratory organs that are sensitive to changes in the water environment. Therefore, we used the liver and gills of Mandarin fish as research materials. The aim of this study was to investigate the effects of short-term artificial feed domestication on the expression of oxidative stress genes and the changes in oxidative-stress-related enzyme activity in the liver and gills of Mandarin fish. We divided the Mandarin fish into two groups for treatment. The control group was fed with live bait continuously for 14 days, while the experimental group was fed with half artificial feed and half live bait from 0 to 7 days (T-7 d), followed by solely artificial feed from 7 to 14 days (T-14 d). The experimental results showed that there was no difference in the body weight, length, and standard growth rate of the Mandarin fish between the two groups of treatments; after two treatments, there were differences in the expression of genes related to oxidative stress in the gills (keap1, kappa, gsta, gstt1, gstk1, SOD, and CAT) and in the liver (GPx, keap1, kappa, gsta, gstt1, gr, and SOD). In the liver, GPx activity and the content of MDA were significantly upregulated after 7 days of domestication, while in the gills, SOD activity was significantly upregulated after 7 days of domestication and GPx activity was significantly downregulated after 14 days of domestication. These results suggest that artificial feed domestication is associated with oxidative stress. Moreover, these results provide experimental basic data for increasing the production of aquaculture feed for Mandarin fish.

Effect of chronic deltamethrin exposure on brain transcriptome and metabolome of juvenile crucian carp.

Deltamethrin (Del), a widely administered pyrethroid insecticide, has been established as a common contaminant of the freshwater environment and detected in many freshwater ecosystems. In this study, we investigated the changes in brain transcriptome and metabolome of crucian carp after exposure to 0.6 µg/L Del for 28 days. Elevated MDA levels and inhibition of SOD activity indicate damage to the antioxidant system. Moreover, a total of 70 differential metabolites (DMs) were identified using the liquid chromatography-mass spectrometry, including 32 upregulated and 38 downregulated DMs in the Del-exposed group. The DMs associated with chronic Del exposure were enriched in steroid hormone biosynthesis, fatty acid metabolism, and glycerophospholipid metabolism for prostaglandin G2, 5-oxoeicosatetraenoic acid, progesterone, androsterone, etiocholanolone, and hydrocortisone. Transcriptomics analysis revealed that chronic Del exposure caused lipid metabolism disorder, endocrine disruption, and proinflammatory immune response by upregulating the pla2g4, cox2, log5, ptgis, lcn, and cbr expression. Importantly, the integrative analysis of transcriptomics and metabolomics indicated that the arachidonic acid metabolism pathway and steroid hormone biosynthesis were decisive processes in the brain tissue of crucian carp after Del exposure. Furthermore, Del exposure perturbed the tight junction, HIF-1 signaling pathway, and thyroid hormone signaling pathway. Overall, transcriptome and metabolome data of our study offer a new insight to assess the risk of chronic Del exposure in fish brains.

The Impact of Niclosamide Exposure on the Activity of Antioxidant Enzymes and the Expression of Glucose and Lipid Metabolism Genes in Black Carp (Mylopharyngodon piceus).

Niclosamide (NIC, 2',5-dichloro-4'-nitrosalicylanilide) is a salicylanilide molluscicide, and the extensive utilization and environmental pollution associated with NIC engender a potential hazard to both human health and the wellbeing of aquatic organisms. However, the mechanism of the chronic toxicity of NIC at environmentally relevant concentrations in terms of oxidative stress, metabolic disorder, and barrier functions in black carp (Mylopharyngodon piceus) is unknown. Therefore, healthy juvenile black carp (M. piceus) (average weight: 38.2 ± 2.5 g) were exposed to NIC at an environmentally realistic concentration (0, 10, and 50 µg/L) for 28 days. The findings of this study indicate that exposure to NIC resulted in reductions in weight gain, decreased activity of antioxidant enzymes, and increased expression of the Nrf2 gene. Furthermore, the liver demonstrated a greater accumulation of NIC than that in the gut and gills, as determined with a chemical analysis. Additionally, NIC exposure led to a significant reduction in ATP content and the activity of Na+/K+-ATPase and Ca2+/Mg2+-ATPase in the gut. Meanwhile, exposure to NIC resulted in a decrease in the liver glucose (Glu) level, gut cholesterol (CHO), and glycogen (Gln) and triglyceride (TG) content in all examined tissues. Conversely, it led to an increase in tissue lactic acid (LA) and acetyl-CoA levels, as well as LDH activity. Furthermore, NIC exposure at environmentally relevant concentrations demonstrated an upregulation in the expression of genes associated with glycolysis, such as PK and GK, while concurrently downregulating the gluconeogenesis gene G6Pase. Additionally, NIC exhibited an upregulation in the expression of genes related to ß-oxidation, such as CPT1 and ACOX, while downregulating genes involved in triglyceride synthesis, including SREBP1, GPAT, FAS, and ACC1. Moreover, NIC facilitated fatty acid transportation through the overexpression of FATP and Fat/cd36. These results suggest that chronic exposure to NIC is associated with oxidative stress, compromised barrier function, and metabolic disorder. Moreover, these results underscore the significance of assessing the potential consequences of NIC for black carp and aquatic environments for aquaculture.

Conventional Anthelmintic Concentration of Deltamethrin Immersion Disorder in the Gill Immune Responses of Crucian Carp.

Current treatment strategies for parasitic infectious diseases in crucian carp primarily rely on chemotherapy. As a commonly used antiparasitic agent, deltamethrin (DEL) may have the potential adverse effects on external mucosa of fish such as gills. In this study, 180 healthy juvenile crucian carp (Carassius auratus) (average weight: 8.8 ± 1.0 g) were randomly divided into three groups for 28 days, which were immersed in 0 µg/L, 0.3 µg/L, and 0.6 µg/L of DEL, respectively. The results of histological analysis revealed that severe hyperplasia in the secondary lamellae of gills was observed, and the number of goblet (mucus-secreting) cells increased significantly after DEL immersion. TUNEL staining indicated that the number of apoptotic cells increased in crucian carp gill. At the molecular level, the mRNA expression analysis revealed significant upregulation of apoptosis (caspase 3, caspase 8, and bax), autophagy (atg5 and beclin-1), and immune response (lzm, muc5, il-6, il-8, il-10, tnfα, ifnγ, tgfß, tlr4, myd88, and nf-kb), whereas tight junction-related genes (occludin and claudin12) were downregulated after DEL immersion, suggesting that DEL immersion altered innate immunity responses and promoted mucus secretion. Moreover, tandem mass tag (TMT)-based proteomics revealed that a total of 428 differentially expressed proteins (DEPs) contained 341 upregulated DEPs and 87 downregulated DEPs with function annotation were identified between the control and DEL groups. Functional analyses revealed that the DEPs were enriched in apoptotic process, phagosome, and lysosome pathways. Additionally, DEL immersion also drove gill microbiota to dysbiosis and an increase in potentially harmful bacteria such as Flavobacterium. Overall, this study showed that DEL elicited shifts in the immune response and changes in the surface microbiota of fish. These results provide new perspectives on the conventional anthelmintic concentration of DEL immersion disorder of the gill immune microenvironment in crucian carp and theoretical support for future optimization of their practical application.

The mitochondrial genome of Heterosentis pseudobagri (Wang & Zhang, 1987) Pichelin & Cribb, 1999 reveals novel aspects of tRNA genes evolution in Acanthocephala.

BACKGROUND: Acanthocephala is a clade of obligate endoparasites whose mitochondrial genomes (mitogenomes) and evolution remain relatively poorly understood. Previous studies reported that atp8 is lacking from acanthocephalan mitogenomes, and that tRNA genes often have nonstandard structures. Heterosentis pseudobagri (Arhythmacanthidae) is an acanthocephalan fish endoparasite for which no molecular data are currently available, and biological information is unavailable in the English language. Furthermore, there are currently no mitogenomes available for Arhythmacanthidae. METHODS: We sequenced its mitogenome and transcriptome, and conducted comparative mitogenomic analyses with almost all available acanthocephalan mitogenomes. RESULTS: The mitogenome had all genes encoded on the same strand and unique gene order in the dataset. Among the 12 protein-coding genes, several genes were highly divergent and annotated with difficulty. Moreover, several tRNA genes could not be identified automatically, so we had to identify them manually via a detailed comparison with orthologues. As common in acanthocephalans, some tRNAs lacked either the TWC arm or the DHU arm, but in several cases, we annotated tRNA genes only on the basis of the conserved narrow central segment comprising the anticodon, while the flanking 5' and 3' ends did not exhibit any resemblance to orthologues and they could not be folded into a tRNA secondary structure. We corroborated that these are not sequencing artefacts by assembling the mitogenome from transcriptomic data. Although this phenomenon was not observed in previous studies, our comparative analyses revealed the existence of highly divergent tRNAs in multiple acanthocephalan lineages. CONCLUSIONS: These findings indicate either that multiple tRNA genes are non-functional or that (some) tRNA genes in (some) acanthocephalans might undergo extensive posttranscriptional tRNA processing which restores them to more conventional structures. It is necessary to sequence mitogenomes from yet unrepresented lineages and further explore the unusual patterns of tRNA evolution in Acanthocephala.

Acute deltamethrin exposure induces oxidative stress, triggers endoplasmic reticulum stress, and impairs hypoxic resistance of crucian carp.

Deltamethrin (Del) has been widely used for effectively controlling ectoparasites of crucian carp and was also strictly prohibited in a hypoxic environment. A previous study indicated that Del exposure causes gill injury in Carassius auratus, which is associated with oxidative stress and endoplasmic reticulum stress (ER stress), but the precise mechanism is not well understood. Here, crucian carp were exposed to Del (0.61, 1.22, 2.44, 4.88 µg/L) for 24 h and then subjected to acute hypoxia challenge (1.0 mg/L) for 24 h. The results revealed that acute exposure to Del notably increased MDA content but markedly decreased CAT activities. Moreover, the T-AOC and SOD activities first increased and then decreased in the 4.88 µg/L Del group. Likewise, the mRNA levels of Nrf2 signaling and its target genes (ho-1, mt, sod, cat, and gpx1) were significantly downregulated in the high concentration exposure groups, while the mRNA levels of keap1 showed the opposite change trend. Meanwhile, Del exposure evoked the PERK-ATF4-CHOP and IRE1 signaling pathways and triggered ER stress in a dose-dependent manner in crucian carp. Importantly, we found that Del exposure significantly decreased the survival rate of crucian carp after hypoxia challenge by reducing oxygen uptake, modifying energy metabolism, and promoting lactate accumulation. Additionally, Del exposure aggravated gill damage and apoptosis under hypoxic stress, which was confirmed by histological assays. Collectively, we inferred that acute exposure to deltamethrin induces oxidative stress and ER stress and impairs hypoxic resistance of crucian carp.

Feature Analysis of Scanning Point Cloud of Structure and Research on Hole Repair Technology Considering Space-Ground Multi-Source 3D Data Acquisition.

As one of the best means of obtaining the geometry information of special shaped structures, point cloud data acquisition can be achieved by laser scanning or photogrammetry. However, there are some differences in the quantity, quality, and information type of point clouds obtained by different methods when collecting point clouds of the same structure, due to differences in sensor mechanisms and collection paths. Thus, this study aimed to combine the complementary advantages of multi-source point cloud data and provide the high-quality basic data required for structure measurement and modeling. Specifically, low-altitude photogrammetry technologies such as hand-held laser scanners (HLS), terrestrial laser scanners (TLS), and unmanned aerial systems (UAS) were adopted to collect point cloud data of the same special-shaped structure in different paths. The advantages and disadvantages of different point cloud acquisition methods of special-shaped structures were analyzed from the perspective of the point cloud acquisition mechanism of different sensors, point cloud data integrity, and single-point geometric characteristics of the point cloud. Additionally, a point cloud void repair technology based on the TLS point cloud was proposed according to the analysis results. Under the premise of unifying the spatial position relationship of the three point clouds, the M3C2 distance algorithm was performed to extract the point clouds with significant spatial position differences in the same area of the structure from the three point clouds. Meanwhile, the single-point geometric feature differences of the multi-source point cloud in the area with the same neighborhood radius was calculated. With the kernel density distribution of the feature difference, the feature points filtered from the HLS point cloud and the TLS point cloud were fused to enrich the number of feature points in the TLS point cloud. In addition, the TLS point cloud voids were located by raster projection, and the point clouds within the void range were extracted, or the closest points were retrieved from the other two heterologous point clouds, to repair the top surface and façade voids of the TLS point cloud. Finally, high-quality basic point cloud data of the special-shaped structure were generated.

Mitochondrial phylogenomics of Acanthocephala: nucleotide alignments produce long-branch attraction artefacts.

BACKGROUND: Classification of the Acanthocephala, a clade of obligate endoparasites, remains unresolved because of insufficiently strong resolution of morphological characters and scarcity of molecular data with a sufficient resolution. Mitochondrial genomes may be a suitable candidate, but they are available for a small number of species and their suitability for the task has not been tested thoroughly. METHODS: Herein, we sequenced the first mitogenome for the large family Rhadinorhynchidae: Micracanthorhynchina dakusuiensis. These are also the first molecular data generated for this entire genus. We conducted a series of phylogenetic analyses using concatenated nucleotides (NUC) and amino acids (AAs) of all 12 protein-coding genes, three different algorithms, and the entire available acanthocephalan mitogenomic dataset. RESULTS: We found evidence for strong compositional heterogeneity in the dataset, and Micracanthorhynchina dakusuiensis exhibited a disproportionately long branch in all analyses. This caused a long-branch attraction artefact (LBA) of M. dakusuiensis resolved at the base of the Echinorhynchida clade when the NUC dataset was used in combination with standard phylogenetic algorithms, maximum likelihood (ML) and Bayesian inference (BI). Both the use of the AA dataset (BI-AAs and ML-AAs) and the CAT-GTR model designed for suppression of LBA (CAT-GTR-AAs and CAT-GTR-NUC) at least partially attenuated this LBA artefact. The results support Illiosentidae as the basal radiation of Echinorhynchida and Rhadinorhynchidae forming a clade with Echinorhynchidae and Pomporhynchidae. The questions of the monophyly of Rhadinorhynchidae and its sister lineage remain unresolved. The order Echinorhynchida was paraphyletic in all of our analyses. CONCLUSIONS: Future studies should take care to attenuate compositional heterogeneity-driven LBA artefacts when applying mitogenomic data to resolve the phylogeny of Acanthocephala.

Test and Analysis of Vegetation Coverage in Open-Pit Phosphate Mining Area around Dianchi Lake Using UAV-VDVI.

This work aimed to detect the vegetation coverage and evaluate the benefits of afforestation and ecological protection. Unmanned aerial vehicle (UAV) aerial survey was adopted to obtain the images of tailings area at Ma'anshan near the Dianchi Lake estuary, so as to construct a high-resolution Digital Orthophoto Map (DOM) and high-density Dense Image Matching (DIM) point cloud. Firstly, the optimal scale was selected for segmentation by considering the terrain. Secondly, the visible-band difference vegetation index (VDVI) of the classified vegetation information of the tail mining area was determined from the index gray histogram, ground class error analysis, and the qualitative and quantitative analysis of the bimodal index. Then, the vegetation information was extracted by combining the random forest (RF) classification algorithm. Finally, the extracted two-dimensional (2D) vegetation information was mapped to the three-dimensional (3D) point cloud, and the redundant data was eliminated. Fractional vegetation cover (FVC) was counted in the way of surface to point and human-machine combination. The experimental results showed that the vegetation information extracted from the 2D image was mapped to the 3D point cloud in the form of surface to point, and the redundant bare ground information was eliminated. The statistical FVC was 36.06%. The field survey suggested that the vegetation information in the turf dam area adjacent to the open phosphate deposit accumulation area research area was sparse. Relevant measures should be taken in the subsequent mining to avoid ecological damage caused by expanded phosphate mining. In general, applying UAV measurement technology and related 2D and 3D products to detect the vegetation coverage in an open phosphate mine area was of practical significance and unique technical advantages.

Landslide Susceptibility Mapping with Integrated SBAS-InSAR Technique: A Case Study of Dongchuan District, Yunnan (China).

Landslide susceptibility maps (LSM) are often used by government departments to carry out land use management and planning, which supports decision makers in urban and infrastructure planning. The accuracy of conventional landslide susceptibility maps is often affected by classification errors. Consequently, they become less reliable, which makes it difficult to meet the needs of decision-makers. Therefore, it is proposed in this paper to reduce classification errors and improve LSM reliability by integrating the Small Baseline Subsets-Interferometric Synthetic Aperture Radar (SBAS-InSAR) technique and LSM. By using the logistic regression model (LR) and the support vector machine model (SVM), experiments were conducted to generate LSM in the Dongchuan district. It was classified into five classes: very high susceptibility, high susceptibility, medium susceptibility, low susceptibility, and very low susceptibility. Then, the surface deformation rate of the Dongchuan area was obtained through the ascending and descending orbit sentinel-1A data from January 2018 to January 2021. To correct the classification errors, the SBAS-InSAR technique was integrated into LSM under the optimal model by constructing the contingency matrix. Finally, the LSMs obtained before and after correction were compared. Moreover, the correction results were validated and analyzed by combining remote sensing images, InSAR deformation results, and field surveys. According to the research results, the susceptibility class of 66,094 classification error cells (59.48 km2) was significantly improved in the LSM after the integration of the SBAS-InSAR correction. The enhanced susceptibility classes and the spectral characteristics of remote sensing images are highly consistent with the trends of InSAR cumulative deformation and the results of field investigation. It is suggested that integrating SBAS-InSAR and LSM is effective in correcting classification errors and further improving the reliability of LSM for landslide prediction. The LSM obtained by using this method plays an important role in guiding local government departments on disaster prevention and mitigation, which is conducive to eliminating the risk of landslides.

Chronic exposure to deltamethrin disrupts intestinal health and intestinal microbiota in juvenile crucian carp.

The indiscriminate use of deltamethrin in agriculture and aquaculture can lead to residues increased in many regions, which poses negative impacts on intestinal health of aquatic organisms. Although the potential toxicity of deltamethrin have recently attracted attention, the comprehensive studies on intestinal injuries after chronic deltamethrin exposure remain poorly understood. Herein, in a 28-day chronic toxicity test, crucian carp expose to different concentrations of deltamethrin (0, 0.3, and 0.6 µg/L) were used as the research object. We found that the morphology changes and increased goblet cells in intestinal tissue, and the extent of tissue injury increased along with the increasing exposure dose of deltamethrin. Additionally, the genes expression of antioxidant activity (Cu/Zn superoxide dismutase (Cu-Zn SOD), glutathione peroxidase 1 (GPX1), and catalase (CAT)), inflammatory response (tumor necrosis factor alpha (TNFα), interferon gamma (IFNγ), and interleukin 1 beta (IL-1ß)), and tight junctions (Claudin 12 (CLDN12), and tight junction protein 1 (ZO-1)) dramatically increased. Meanwhile, the apoptosis and autophagy process were triggered through caspase-9 cascade and autophagy related 5 (ATG5)- autophagy related 12 (ATG12) conjugate. Besides, chronic deltamethrin exposure increased the amount of Proteobacteria and Verrucomicrobiota, while decreased Fusobacteriota abundance, resulting in intestinal microbiota function disorders. In summary, our results highlight that chronic exposure to deltamethrin cause serious intestinal toxicity and results in physiological changes and intestinal flora disturbances.

Two new species of Bacillidium (Microsporidia) from coelomocytes of Branchiura sowerbyi (Oligochaeta: Naididae) in China.

Bacillidium spp. exclusively infect oligochaetes and these microsporidian pathogens are typically characterized by their rod-shaped spores. Seven Bacillidium spp. are presently reported from different organs of oligochaetes. Here, we describe two new Bacillidium species, Bacillidium sinensis n. sp. and Bacillidium branchilis n. sp., from coelomocytes of Branchiura sowerbyi. This is the first report of Bacillidium spp. in oligochaetes from China. Both species of Bacillidium elicit the formations of opaque xenoma-like lesions in coelomocytes of the host. A diplokaryotic nucleus occurs in all life stages of these two new Bacillidium species. Mature spores of B. sinensis are 15.9 ± 0.6 (14.7-17.1) µm long (average ± standard error, range, n = 50) and 2.5 ± 0.1 (2.3-2.7) µm wide in fresh preparations. A new type of exospore (sixteen-layered exospore) is discovered from B. sinensis n. sp. which is distinctly different from B. branchilis n. sp., and other Bacillidium spp. (double-layered exospore) reported previously. These two Bacillidium species are morphologically distinguished from each other and all Bacillidium spp. described previously in terms of hosts, infection sites, spore size, spore wall or polar filament thickness. BLASTn searches indicated that these two new microsporidian parasites are surprisingly most similar to Janacekia tainanus (94.76% for B. sinensis and 90. 2% for B. branchilis) isolated from the fat body of midge larva (Kiefferulus tainanus). Phylogenetic analysis demonstrates that the two novel taxons cluster with J. debaisieuxi, J. tainanus, and Bacillidium sp. within the Jirovecia-Bacillidium-Janacekia clade. Other available 18S rRNA gene sequences for microsporidia that infect oligochaetes include J. sinensis, B. vesiculoformis, Neoflabelliforma aurantiae, and Bacillidium sp., but these do not form a single cluster with B. sinensis and B. branchilis, but are instead dispersed through the clade. Based on the ultrastructural features and molecular characteristics, two new species within the genus Bacillidium, B. sinensis n. sp. and B. branchilis n. sp., are designated.

Naidispora caidianensis n. gen. n. sp. infecting coelomocytes of oligochaete Branchiura sowerbyi (Oligochaeta: Naididae) in China.

A new microsporidian parasite, Naidispora caidianensis n. gen. n. sp. was found in coelomocytes of oligochaete Branchiura sowerbyi Beddard, 1892 from Wuhan city, Hubei Province, China. Opaque, hypertrophied coelomocytes (0.17-0.24 mm in diameter) depicted clinical signs of infection. Electron microscopy revealed a microsporidian with monokaryotic life stages. Rounded uninucleate meronts subsequently transformed into multinucleate merogonial plasmodia with masses of electron-dense projections on their plasma membrane. Electron-dense sporogonial plasmodia separated into uninucleate sporonts through rosette-like budding, and further developed into sporoblasts, enclosed by a sporophorous vesicle. Uninucleate mature spores were pyriform, measured 4.1 ± 0.1 (3.9-4.3) µm × 2.1 ± 0.07 (1.9-2.2) µm (average ± SE, range, n = 50), and contained a mushroom-like anchoring disk, bipartite polaroplast, electron-lucent posterior vacuole, trilaminar spore wall and 12-14 turns of an isofilar polar filament arranged in 2-3 ranks. The 1542 bp sequence from B. soweryi showed the highest similarity (below 80 %) with Nematocenator marisprofundi (accession number: JX463178). SSU rRNA gene-based phylogenetic analysis demonstrated that the novel taxon formed an independent clade from known microsporidian parasites. Based on the ultrastructural features and SSU rRNA gene sequence, we propose the establishment of a novel genus (Naidispora n. gen.) and species (Naidispora caidianensis n. sp.) to contain this parasite.

Histopathology and transcriptome analysis reveals the gills injury and immunotoxicity in gibel carp following acute deltamethrin exposure.

More and more evidences proved that deltamethrin (Del) exposure induced adverse effects and damaged immune function to the aquatic animals in the parasite killing process with increasing insecticide application. However, little is currently known of the negative effect on mucosal immunity, especially in gills tissue. Therefore, this study was aimed to reveal the tissue injury and immunotoxicity in the gill of gibel carp following acute deltamethrin exposure. The LC50 of deltamethrin on gibel carp at 96 h was determined to be 6.194 µg/L, and then juvenile gibel carp (Carassius auratus gibelio) (8.8 ± 1.0 g) were exposed to four Del exposure groups (0.61, 1.22, 2.44, and 4.88 µg/L) for 12 h and 24 h. We measured the lysozyme (LYZ) contents and myeloperoxidase (MPO) activities and found that with increased concentration of Del exposure, the LYZ contents were found to increase in the 1.22 µg/L Del group initially significantly and then gradually significantly decrease in the 4.88 µg/L Del group. And the activities of MPO were significantly lifted in a dose-dependent manner. The histological analysis showed that Del exposure caused serious desquamation and necrosis in the surface of epithelial cells, accompanied by interlamellar cellular mass degenerative. In addition, the mucous cells were significantly decreased in the high Del concentration group (2.44 µg/L and 4.88 µg/L Del group) by AB-PAS staining. Additionally, totally 2857 DEGs (including 1624 up-regulated and 1233 down-regulated genes) were identified between the control group and 4.88 µg/L Del exposure group using transcriptional analysis. Among these, some genes involved in innate immune molecules, complement activation, apoptosis-related molecules, cytokine, and adaptive immune molecules, were also down-regulated. Importantly, we found immune system process and tumor necrosis factor receptor (superfamily) binding pathways were downregulated based on the GO and KEGG enrichment analysis. Meanwhile, we detected the expression of pro-inflammatory cytokines (TNF-α, IFN-γ, IL-1ß, and IL-8), anti-inflammatory cytokines (TGF-ß), LYZ, IgM, and Hsp70 in the gills tissue at 12 h and 24 h after Del exposure, which were consistent with our sequencing results. Collectively, these results demonstrated that the gills injury and immunotoxicity were induced by Del exposure and provided novel insight for explaining to some extent why Del-exposure fish are more susceptible to concurrent or secondary viral or bacterial infections.

Studies on Three-Dimensional (3D) Accuracy Optimization and Repeatability of UAV in Complex Pit-Rim Landforms As Assisted by Oblique Imaging and RTK Positioning.

Unmanned Aerial Vehicles (UAVs) are a novel technology for landform investigations, monitoring, as well as evolution analyses of long-term repeated observation. However, impacted by the sophisticated topographic environment, fluctuating terrain and incomplete field observations, significant differences have been found between 3D measurement accuracy and the Digital Surface Model (DSM). In this study, the DJI Phantom 4 RTK UAV was adopted to capture images of complex pit-rim landforms with significant elevation undulations. A repeated observation data acquisition scheme was proposed for a small amount of oblique-view imaging, while an ortho-view observation was conducted. Subsequently, the 3D scenes and DSMs were formed by employing Structure from Motion (SfM) and Multi-View Stereo (MVS) algorithms. Moreover, a comparison and 3D measurement accuracy analysis were conducted based on the internal and external precision by exploiting checkpoint and DSM of Difference (DoD) error analysis methods. As indicated by the results, the 3D scene plane for two imaging types could reach an accuracy of centimeters, whereas the elevation accuracy of the orthophoto dataset alone could only reach the decimeters (0.3049 m). However, only 6.30% of the total image number of oblique images was required to improve the elevation accuracy by one order of magnitude (0.0942 m). (2) An insignificant variation in internal accuracy was reported in oblique imaging-assisted datasets. In particular, SfM-MVS technology exhibited high reproducibility for repeated observations. By changing the number and position of oblique images, the external precision was able to increase effectively, the elevation error distribution was improved to become more concentrated and stable. Accordingly, a repeated observation method only including a few oblique images has been proposed and demonstrated in this study, which could optimize the elevation and improve the accuracy. The research results could provide practical and effective technology reference strategies for geomorphological surveys and repeated observation analyses in sophisticated mountain environments.

Characterization and comparative analysis of the complete mitochondrial genome of Azygia hwangtsiyui Tsin, 1933 (Digenea), the first for a member of the family Azygiidae.

Azygia hwangtsiyui (Trematoda, Azygiidae), a neglected parasite of predatory fishes, is little-known in terms of its molecular epidemiology, population ecology and phylogenetic study. In the present study, the complete mitochondrial genome of A. hwangtsiyui was sequenced and characterized: it is a 13,973 bp circular DNA molecule and encodes 36 genes (12 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes) as well as two non-coding regions. The A+T content of the A. hwangtsiyui mitogenome is 59.6% and displays a remarkable bias in nucleotide composition with a negative AT skew (-0.437) and a positive GC skew (0.408). Phylogenetic analysis based on concatenated amino acid sequences of twelve protein-coding genes reveals that A. hwangtsiyui is placed in a separate clade, suggesting that it has no close relationship with any other trematode family. This is the first characterization of the A. hwangtsiyui mitogenome, and the first reported mitogenome of the family Azygiidae. These novel datasets of the A. hwangtsiyui mt genome represent a meaningful resource for the development of mitochondrial markers for the identification, diagnostics, taxonomy, homology and phylogenetic relationships of trematodes.

Complete mitochodrial genome of Vanmanenia pingchowensis (Cypriniformes, Cyprinidae).

The complete mitochondrial genome of Vanmanenia pingchowensis was determined in this study. The gene composition, arrangement and transcriptional orientation in V. pingchowensis mitogenome were identical to most vertebrates. The complete mitogenome of V. pingchowensis was 15,560 bp in size with 13 protein coding genes, 22 tRNA genes, and a control region. Two start codon patterns and three stop codon patterns were found in protein-coding genes. Only the tRNA-Ser2 could not fold into a typical clover-leaf secondary structure for lacking the dihydrouridine arm. Sequences alignment results suggest that the complete mitogenome of V. pingchowensis is an efficient tool to study molecular phylogenetics, biogeography and adaptive evolution of this lineage.

Complete mitochodrial genome of Elopichthys bambusa (Cypriniformes, Cyprinidae).

The complete mitochondrial genome of Elopichthys bambusa was determined in this study. The gene composition, arrangement and transcriptional orientation in E. bambusa mitogenome were identical to most vertebrates. Two start codon patterns (ATG and GTG) and three stop codon patterns (TAG, TAA and T) were found in protein-coding genes. Only the tRNA-Ser2 could not fold into a typical clover-leaf secondary structure for lacking the dihydrouridine arm. Sequences alignment results suggest that the complete mitogenome of E. bambusa is an efficient tool to check species identification by comparing different gene sequences.