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The effective remission of acute respiratory distress syndrome- (ARDS-) caused pulmonary fibrosis determines the recovery of lung function. Inositol can relieve lung injuries induced by ARDS. However, the mechanism of myo-inositol in the development of ARDS is unclear, which limits its use in the clinic. We explored the role and mechanism of myo-inositol in the development of ARDS by using an in vitro lipopolysaccharide- (LPS-) established alveolar epithelial cell inflammation model and an in vivo ARDS mouse model. Our results showed that inositol can alleviate the progression of pulmonary fibrosis. More significantly, we found that inositol can induce autophagy to inhibit the progression pulmonary fibrosis caused by ARDS. In order to explore the core regulators of ARDS affected by inositol, mRNA-seq sequencing was performed. Those results showed that transcription factor HIF-1α can regulate the expression of SLUG, which in turn can regulate the key gene E-Cadherin involved in cell epithelial-mesenchymal transition (EMT) as well as N-cadherin expression, and both were regulated by inositol. Our results suggest that inositol activates autophagy to inhibit EMT progression induced by the HIF-1α/SLUG signaling pathway in ARDS, and thereby alleviates pulmonary fibrosis.

There is considerable confusion concerning the relationships among species of Sarcocystis found in donkeys and horses. Here, we describe a Sarcocystis species in Chinese donkeys (Equus asinus). Sarcocysts were found in 12 of 32 (37.5%) adult donkeys. By light microscopy, they were divided into two types, thin-walled and thick-walled. The thin-walled were macroscopic (up to 320 μm wide) and had short club-like protrusions (up to 2.7 μm long); the thick-walled were microscopic (up to 135 μm wide) and had villar protrusions (up to 5.4 μm long). Ultrastructures of the two types exhibited similar morphological characteristics, including bundled microtubules in the core of the villar protrusions penetrating diagonally into the ground substance, similar to wall type 11c. Three genetic markers, 18S rDNA, 28S rDNA, and mitochondrial cox1, obtained from the two morphotypes were sequenced and analyzed. The sequences of the three loci in the two morphotypes presented high intraspecific similarities of 97.2-99.5%, 97.8-99.6% and 99.0 - 99.9%, respectively. The most similar sequences in GenBank to the newly obtained 18S rDNA, 28S rDNA and cox1 sequences were those of Sarcocystis spp. in horses, with similarities of 90.0 - 97.5%, 94.7 - 95.1%, and 82.6 - 84.5%, respectively. Phylogenetic analysis using the three genetic markers indicated that the Sarcocystis sp. in donkeys formed an individual clade most closely related to a clade encompassing Sarcocystis spp. in horses. Further studies are needed for taxonomic identification of sarcocysts in donkeys because the Sarcocystis species in donkeys and horses are not successfully cross transmissible despite morphological similarities.

Macrogametocyte stage antigens reportedly provide protective immunity against coccidiosis in poultry. This study was designed to evaluate the ability of a purified recombinant protein from Eimeria necatrix gametocytes (rEnGAM22) to stimulate immunity against experimental infection with sporulated E. necatrix oocysts. The immunogenicity of the recombinant protein was studied in chickens by subcutaneous injection of 25, 50, or 75 µg of the protein with Freund’s adjuvant. Vaccine efficacy was assessed after oral parasite challenge by fecal oocyst output, lesion scores, body weight gain, serum antibodies, and cytokine responses. Chickens vaccinated with 50 µg of rEnGAM22 and challenged with sporulated E. necatrix oocysts showed reduced fecal oocyst shedding and lesion scores compared with other immunized groups and the infected control group, with the exception of the live oocyst group. There was no difference in body weight between the immunized groups and the infected control group. Furthermore, rEnGAM22 also stimulated higher production of anti-rEnGAM22 serum antibodies 7 days after secondary immunization, especially with 50 µg of rEnGAM22. Serum levels of interleukin (IL)-2, IL-4, IL10, and interferon (IFN)-γ also showed the greatest immune response from 50 µg of rEnGAM22, and IL-2 and IL-4 responses were greater than those of IL-10 and IFN-γ. These results indicated that rEnGAM22 protected against E. necatrix infection to some degree and may potentially be used to develop a recombinant subunit vaccine against coccidiosis.

Background: Data on the genus Sarcocystis in waterfowls are very limited. To the best of our knowledge, the species of Sarcocystis have previously never been recorded in ducks in China. Methods: Leg muscles were obtained from 26 domestic ducks Anas platyrhynchos in 2021 in China. Sarcocysts were observed using light microscopy (LM) and transmission electron microscopy (TEM). Individual sarcocysts from different hosts were selected for DNA extraction, and three genetic markers, namely, 18S ribosomal DNA (18S rDNA), 28S ribosomal DNA (28S rDNA), and mitochondrial cytochrome oxidase subunit 1 (cox1), were amplified, sequenced, and analyzed. Results: Sarcocysts were found in 3 of the 28 (10.7%) domestic ducks. Under LM, the sarcocysts were microscopic and had brush-like villar protrusions (vps), measuring 3.8–4.3 μm-long. Ultrastructurally, the sarcocyst wall contained numerous lancet- or leaf-like vp. Each vps narrowed in the stalk and contained bundled microtubules extended into the ground substance. Comparison of the newly obtained sequences with those previously deposited in GenBank indicated that the most similar sequences were those of S. halieti ex Phalacrocorax carbo or Sturnus vulgaris,and S. calchasi ex Columba livia, with an identity of 99.1% at 18S rDNA, S. wenzeli ex Gallus gallus, with an identity of 95.9–96.0% identity at the 28S rDNA, and S. speeri ex opossum, with an identity of 98.2% at the mitochondrial cox1. Additionally, at the three loci, the newly obtained sequences unambiguously differed from those of Sarcocystis spp. obtained from Anseriformes birds. Phylogenetic analysis based on the three loci placed the parasite within a group containing Sarcocystisspp. that utilize birds or dogs as intermediate hosts. The parasite was proposed as a new species, named Sarcocystis platyrhynchosi. Conclusions:Sarcocysts were recorded in domestic ducks in China for the first time. Morphologically and molecularly, the sarcocysts remarkably differed from those of Sarcocystis spp. previously reported in different intermediate hosts, and S. platyrhynchosi was proposed for the new species. Depending on the limited date provided for Sarcocystisspp. in Anseriformes birds, there is confusion concerning the relationships of the morphologically similar species of Sarcocystisin Anseriformes birds. Therefore, more research on these closely related species of Sarcocystis needs to be undertaken in the future.

Shallow soil with low water availability, poor nutrition, and limited living space is the key limiting factor of plant growth and reproduction in vulnerable karst regions. As pioneers adapted to these areas, little is known about the reproductive responses of annual herbs to these limited soil resources. In this study, we, therefore, investigated the reproductive strategies (“fight or flight”) of two annual Asteraceae herbs (Xanthium sibiricum and Bidens pilosa) growing in the harsh karst soil environment, by comprehensively considering biomass allocation, reproductive phenology, and seed production. X. sibiricum and B. pilosa were grown under three soil depths and three water availability levels in a full two-way randomized block design, then biomass allocation, initial flowering time, fruiting stage, life span and seed number and mass were examined. In X. sibiricum, reproductive allocation decreased with decreasing water availability, soil depth, and water availability + soil depth. Meanwhile, the initial flowering time, fruiting stage and life span were all delayed with moderate decreases in resources. However, in B. pilosa, reproductive allocation increased with decreasing soil resources, while initial flowering time, fruiting stage and life span remained unchanged under moderate resource decline. Under severe resource decline, both species showed a reduction in fruiting stage and life span. These results support the hypothesis that X. sibiricum tends to adopt a “fight” reproductive strategy in response to declining resources via changes in biomass allocation, reproductive phenology and seed production. In contrast, B. pilosa tends to adopt a “flight” strategy in terms of biomass allocation, while maintaining a relatively stable reproductive phenology and seed size to cope with the harsh living conditions. However, neither species coped well under severe declines in water availability, nutrition and living space. These findings help our understanding of how herbaceous plants can adjust their reproductive strategies to support their regeneration in harsh karst soil environments.

Purpose Recent clinical trials with agents targeting immune checkpoint pathway have emerged as an important therapeutic approach for a broad range of cancer types. Resveratrol has been shown to possess cancer preventive and therapeutic effects and has potential to be chemotherapeutic agent/adjuvant. Here, we assessed the effect of resveratrol on immune checkpoint pathways. Methods The expression patterns of Wnt components and PD-L1 were examined by Western blot, Chromatin immunoprecipitation (ChIP) was used for analysis of DNA–protein interaction, the promoter activity was determined by luciferase reporter assay, apoptosis was analyzed by flow cytometry and the ability of the resveratrol to modulate T cell function was assessed in a co-culture system. Results Although the dose-, and cell-type dependent effects of resveratrol on PD-L1 expression have been reported, we show here that resveratrol dose-dependently upregulates PD-L1 expression at the range of pharmacologic-achievable concentrations in lung cancer cells and that is essential for suppression of T-cell-mediated immune response. We also found that Wnt pathway is critical for mediating resveratrol-induced PD-L1 upregulation. Mechanistically, resveratrol activates SirT1 deacetylase to deacetylate and stabilize transcriptional factor Snail. Snail in turn inhibits transcription of Axin2, which leads in disassembly of destruction complex and enhanced binding of β-catenin/TCF to PD-L1 promoter. Conclusion We conclude that resveratrol is capable to suppress anti-tumor immunity by controlling mainly PD-L1 expression. This finding will extend the understanding of resveratrol in regulation of tumor immunity and is relevant to the debate on resveratrol supplements for lung cancer patients.

Background Sepsis in children is one of the main causes of death in pediatric intensive care units (PICUs); however, the pathogenesis of sepsis is not fully clear. Previous studies revealed that many genetic variations were related to sepsis susceptibility. A long non-coding RNA SOX2 overlapping transcript (SOX2OT) may play a role in mitochondrial homeostasis and antioxidative activity, but the relationship between the lncRNA SOX2OT polymorphism and sepsis susceptibility has not been reported. Methods In this study, 474 pediatric sepsis patients and 678 healthy controls were recruited from southern China. After genotyping, the strength of the associations was evaluated through odds ratios (ORs) and 95% confidence intervals (CIs). Results The SOX2OT rs9839776 T allele was associated with decreased susceptibility to sepsis in southern Chinese children (TT/CT vs CC adjusted OR = 0.778, 95% CI = 0.610-0.992; P = 0.0431). Moreover, the difference in susceptibility was greater in children of age >60 months (adjusted OR = 0.458, 95% CI = 0.234-0.896; P = 0.0225), survivors (adjusted OR = 0.758, 95% CI = 0.585-0.972; P = 0.0358), males (adjusted OR = 0.655, 95% CI = 0.479-0.894; P = 0.0077) and the sepsis subgroup (adjusted OR = 0.548, 95% CI = 0.343-0.876; P = 0.0120). Conclusion The rs9839776 T allele may contribute to decreased sepsis risk in Chinese children. Future studies with a larger sample size are needed to verify these results.

There is considerable confusion concerning the relationships among species of Sarcocystis found in donkeys and horses. Here, sarcocysts obtained from donkeys (Equus asinus) were morphologically and molecularly characterized. Sarcocysts were diagnosed in 12 of 32 (37.5%) Chinese donkeys and could be divided into two types, thin-walled and thick-walled, with the aid of light microscopy (LM). The thin-walled sarcocysts were macroscopic (up to 4856 × 320 µm in size) and had short club-like protrusions of up to 2.7 µm; the thick-walled sarcocysts were microscopic (up to 3750 × 135 µm in size) and had villar protrusions of up to 5.4 µm. The ultrastructures of the two types exhibited highly similar morphological characteristics, including bundled microtubules in the core of the villar protrusions penetrating diagonally into the ground substance, similar to wall type 11c. Three genetic markers, 18S rDNA, 28S rDNA, and mitochondrial cox1, were sequenced and analyzed in the two types. The three genetic markers presented high intraspecific similarity between the two types, ranging from 97.2–99.5%, 97.8–99.6% and 99.0 − 99.9%, respectively. The comparison of these sequences with those of Sarcocystis spp. previously deposited in GenBank showed that the newly obtained sequences of 18S rDNA, 28S rDNA, and mitochondrial cox1 presented identities of 90.0 − 97.5%, 94.7 − 95.1% and 82.6 − 84.5%, respectively, with those of S. bertrami and S. fayeri obtained from horses. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) based on the mitochondrial cox1 sequences of donkey and horse sarcocysts revealed three fragments (196, 243 and 641 bp) and two fragments (416 and 644 bp), respectively, upon digestion with the restriction enzymes EcoRI and HinfI. Phylogenetic analysis based on 18S rDNA or mitochondrial cox1 sequences revealed that donkey sarcocysts formed an individual clade with Sarcocystis spp. obtained from horses. Based on the divergence of the three sarcocyst genetic markers, especially mitochondrial cox1, between donkeys and horses, the Sarcocystis sp. sarcocysts in donkeys were different with those obtained from horses. The cross-infection of Sarcocystis between donkeys and horses needs to be attempted for further confirmation in the future.

There is considerable confusion concerning the relationships among species of Sarcocystis found in donkeys and horses. Here, sarcocysts obtained from donkeys (Eqqus asinus) were morphologically and molecularly characterized. Sarcocysts were diagnosed in 12 of 32 (37.5%) Chinese donkeys and could be divided into two forms, thin-walled and thick-walled, with the aid of light microscopy (LM). The thin-walled sarcocysts were macroscopic (up to 4856 × 320 μm in size) and had short club-like protrusions of up to 2.7 μm; the thick-walled sarcocysts were microscopic (up to 3750 × 135 μm in size) and had hair-like protrusions of up to 5.4 μm. The ultrastructures of the two forms exhibited highly similar morphological characteristics, including bundled microtubules in the core of the villus protrusions penetrating diagonally into the ground substance, similar to wall type 11. Three genetic markers, 18S rDNA, 28S rDNA, and mitochondrial cox1, were sequenced and analyzed in the two forms. The three genetic markers presented high intraspecific similarity between the two forms, ranging from 97.2–99.5% (average 97.8%), 97.8-99.6% (average 98.4%) and 99.0−99.9% (average 99.4%), respectively. The comparison of these sequences with those of Sarcocystis spp. previously deposited in GenBank showed that the newly obtained sequences of 18S rDNA, 28S rDNA, and mitochondrial cox1 presented identities of 90.0−97.5% (average 94.7%), 94.7−95.1% (average 94.9%) and 82.6−84.5% (average 83.4%), respectively, with those of S. bertrami/fayeri obtained from horses. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) based on the mitochondrial cox1 sequences of donkey and horse sarcocysts revealed three fragments (196, 243 and 641 bp) and two fragments (416 and 644 bp), respectively, upon digestion with the restriction enzymes HinifI and EcorRI. Phylogenetic analysis based on 18S rDNA or mitochondrial cox1 sequences revealed that donkey sarcocysts formed an individual clade that clustered with Sarcocystis spp. obtained from horses. Based on the divergence of the three sarcocyst genetic markers, especially mitochondrial cox1, between donkeys and horses, Sarcocystis asinus should be resurrected and regarded as a valid species in donkeys. The cross-infection of Sarcocystis between donkeys and horses needs to be attempted for further confirmation in the future.

Background: Diarrhea is one of the important diseases of sucking goats and seriously affects the survival rate of lambs. Many studies have been reported on the etiology of goat diarrhea, but the available information on the intestinal flora disorder in diarrheic goats was limited. Methods: To understand the impact of the gut microbes on goat health and to determine the changes in the intestinal flora of sucking goats with diarrhea, 13 fecal samples from 9 diarrheic lambs and 4 healthy lambs were collected and submitted to metagenomics analysis. Results: The data show that the intestinal flora of diarrheic goats was not only significant changed in the bacterial species, but also displayed abundances decrease in probiotics, such as Butyricicoccus pullicaecorum, Ruminococcus gnavus, Bacteroides fragilis, etc. Many harmful bacteria harboring in the intestinal flora and pathogenic bacteria invading from in vitro should be important bacteria that cause intestinal flora disorder. The most dominant harmful bacteria include Staphylococcus aureus, Lactococcus garvieae, Helicobacter pylori, Escherichia coli, Firmicutes bacterium, Bacteroides plebeius, etc. Conclusion: The intestine flora of sucking goats with diarrhea changed significantly in both species and abundance. Disorder of intestinal flora should be one of the important causes of diarrhea in sucking goats.

BackgroundToxoplasma gondiiis an obligate intracellular protozoan parasite that can cause a geographically widespread zoonosis. Our previous splenocyte microRNA profile analyses of pig infected withT. gondiirevealed that the coordination of a large number of miRNAs regulates the host immune response during infection. However, the functions of other miRNAs involved in the immune regulation duringT. gondiiinfection are not yet known.MethodsClustering analysis was performed byK-means, self-organizing map (SOM), and hierarchical clustering to obtain miRNA groups with the similar expression patterns. Then, the target genes of the miRNA group in each subcluster were further analyzed for functional enrichment by Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Reactome pathway to recognize the key signaling molecules and the regulatory signatures of the innate and adaptive immune responses of the host duringT. gondiiinfection.ResultsA total of 252 miRNAs were successfully divided into 22 subclusters byK-means clustering (designated as K1–K22), 29 subclusters by SOM clustering (designated as SOM1–SOM29), and six subclusters by hierarchical clustering (designated as H1–H6) based on their dynamic expression levels in the different infection stages. A total of 634, 660, and 477 GO terms, 15, 26, and 14 KEGG pathways, and 16, 15, and 7 Reactome pathways were significantly enriched byK-means, SOM, and hierarchical clustering, respectively. Of note, up to 22 miRNAs mainly showing downregulated expression at 50 days post-infection (dpi) were grouped into one subcluster (namely subcluster H3-K17-SOM1) through the three algorithms. Functional analysis revealed that a large group of immunomodulatory signaling molecules were controlled by the different miRNA groups to regulate multiple immune processes, for instance, IL-1-mediated cellular response and Th1/Th2 cell differentiation partly depending on Notch signaling transduction for subclusters K1 and K2, innate immune response involved in neutrophil degranulation and TLR4 cascade signaling for subcluster K15, B cell activation for subclusters SOM17, SOM1, and SOM25, leukocyte migration, and chemokine activity for subcluster SOM9, cytokine–cytokine receptor interaction for subcluster H2, and interleukin production, chemotaxis of immune cells, chemokine signaling pathway, and C-type lectin receptor signaling pathway for subcluster H3-K17-SOM1.ConclusionsCluster analysis of splenocyte microRNAs in the pig revealed key regulatory properties of subcluster miRNA molecules and important features in the immune regulation induced by acute and chronicT. gondiiinfection. These results contribute new insight into the identification of physiological immune responses and maintenance of tolerance in pig spleen tissues duringT. gondiiinfection.Graphical Abstract

Background Eimeria necatrix is one of the most pathogenic parasites, causing high mortality in chickens. Although its genome sequence has been published, the sequences and complete structures of its mRNA transcripts remain unclear, limiting exploration of novel biomarkers, drug targets and genetic functions in E. necatrix. Methods Second-generation merozoites (MZ-2) of E. necatrix were collected using Percoll density gradients, and high-quality RNA was extracted from them. Single-molecule real-time (SMRT) sequencing and Illumina sequencing were combined to generate the transcripts of MZ-2. Combined with the SMRT sequencing data of sporozoites (SZ) collected in our previous study, the transcriptome and transcript structures of E. necatrix were studied. Results SMRT sequencing yielded 21,923 consensus isoforms in MZ-2. A total of 17,151 novel isoforms of known genes and 3918 isoforms of novel genes were successfully identified. We also identified 2752 (SZ) and 3255 (MZ-2) alternative splicing (AS) events, 1705 (SZ) and 1874 (MZ-2) genes with alternative polyadenylation (APA) sites, 4019 (SZ) and 2588 (MZ-2) fusion transcripts, 159 (SZ) and 84 (MZ-2) putative transcription factors (TFs) and 3581 (SZ) and 2039 (MZ-2) long non-coding RNAs (lncRNAs). To validate fusion transcripts, reverse transcription-PCR was performed on 16 candidates, with an accuracy reaching up to 87.5%. Sanger sequencing of the PCR products further confirmed the authenticity of chimeric transcripts. Comparative analysis of transcript structures revealed a total of 3710 consensus isoforms, 815 AS events, 1139 genes with APA sites, 20 putative TFs and 352 lncRNAs in both SZ and MZ-2. Conclusions We obtained many long-read isoforms in E. necatrix SZ and MZ-2, from which a series of lncRNAs, AS events, APA events and fusion transcripts were identified. Information on TFs will improve understanding of transcriptional regulation, and fusion event data will greatly improve draft versions of gene models in E. necatrix. This information offers insights into the mechanisms governing the development of E. necatrix and will aid in the development of novel strategies for coccidiosis control. Graphical Abstract

Background: There are limited data on Sarcocystis in insectivores. The Asian gray shrew, Crocidura attenuata, is one of the most common species of insectivores in the family Soricidae distributed in South Asia and Southeast Asia. To date, Sarcocystis has never been recorded in this host.Methods: Tissues from 42 Asian gray shrews were collected in China in 2017 and 2018. Sarcocysts were observed using light (LM) and transmission electron microscopy (TEM). To complete the parasite life cycle, muscle tissues of the host infected with sarcocysts were force-fed to two beauty rat snakes, Elaphe taeniura. Individual sarcocysts from different Asian gray shrews and oocysts/sporocysts isolated from the small intestines and feces of the experimental snakes were selected for DNA extraction, and seven genetic markers, including two nuclear loci (18S rDNA and ITS1), three mitochondrial genes (cox1, cox3 and cytb), and two apicoplastic genes (rpoB and clpC), were amplified, sequenced and analyzed.Results: Sarcocysts were found in 17 of 42 (40.5%) Asian gray shrews. Under LM, the microscopic sarcocysts were exhibited saw-tooth-like protrusions measuring 3.3–4.5 μm. Ultrastructurally, the sarcocyst wall contained numerous lancet- or leaf-like villous protrusions, similar to type 9h. The experimental beauty rat snakes shed oocysts/sporcysts measuring 11.9–16.7 × 9.2–10.6 μm with a prepatent period of 10 to 11 days. Comparing these sequences with those previously deposited in GenBank revealed that the 18S rDNA sequences and cox1 sequences shared the highest similarity with those of S. scandentiborneensis recorded in tree shrews, Tuaia minor and T. tana (i.e., 97.6–98.3% and 100% identity, respectively). Phylogenetic analysis based on 18S rDNA, ITS1 or cox1 sequences revealed that this parasite formed an independent clade with Sarcocystis spp. that utilize small animals as intermediate hosts and snakes as the known or presumed definitive host. On the basis of morphological and molecular characteristics and host specificity, the parasite was proposed as a new species, named S. attenuati.Conclusions: Sarcocysts were recorded in Asian gray shrews for the first time. The sarcocysts were characterized morphologically and molecularly. The 18S rDNA and cox1 sequences of S. attenuati, named in the present study, shared the highest identities with those of S. scandentiborneensis. However, the sarcocysts of the two species of Sarcocystis were quite different under LM and TEM. Based on experimental infection, beauty rat snakes have been proven to be a definitive host of S. attenuati. As more species of Sarcocystis from insectivores and other small mammals are properly morphologically and molecularly characterized, we may gain a better understanding of the biodiversity, host specificity and evolution of Sarcocystis in the future.

Aims The mechanism by which species diversity drives productivity in different ecosystems is controversial, possibly due to the confounding effects of key environmental variables. Karst ecosystems are fragile and are at great risk of species loss. In these ecosystems, soil depth is a key driver of community diversity and productivity. However, the influence of soil depth on the relationship between species diversity and productivity in karst ecosystems remains unclear.Methods We established artificial karst herbaceous communities with different soil depths and species richness levels and determined how two biodiversity effects—complementarity effect (effect of positive interactions among species) and selection effect (effect due to dominance of productive species)—contributed to productivity.Results Soil depth, species diversity, and different species combinations were significant predictors of productivity. Species diversity significantly positively affected productivity at all three soil depths, with the effect size of species diversity being the greatest in medium-depth soil. Net diversity effects were greater than 0 in all multi-species communities, indicating that complementarity and selection effects both positively influenced productivity. However, complementarity effect had a greater contribution to productivity than selection effect in all multi-species communities. Furthermore, the contribution of complementarity effect increased with increasing soil depth, while that of selection effect decreased.Conclusions Soil depth influences the relationship between species diversity and productivity in karst herbaceous communities. Furthermore, complementarity effect is the major mechanism by which diversity increases community productivity, particularly in shallow soil. Therefore, environmental variables should be seriously considered when studying the relationship between species diversity and function in other ecosystems.

Soil pH varies by several units among ecosystems. While soil pH is known to be a key driver of plant species composition, we still have a poor understanding of how it affects carbon cycling processes. For instance, soil pH, or its associated chemistry in terms of base cations and organic acids, may affect decomposition rates of dead matter directly, by controlling decomposer composition and activity and, indirectly, by controlling the traits of the plant species and thereby the afterlife effects of those traits on litter decomposition. Leaf and litter pH may play a role in this control. Based on the very limited empirical data available, we hypothesized that variation in species traits including leaf (litter) pH, within and between ecosystems contrasting in soil pH, would have stronger effects on leaf litter decomposition rates than variation in soil chemistry would. We tested this hypothesis by carrying out a ‘common garden’ litterbed experiment in subtropical SW China, in which leaf litters of the 30 predominant plant species from mid‐successional forest on acidic sandstone (soil pH around 4.0) and calcareous soil (pH around 7.5), respectively, were incubated and their decomposition rates measured over two harvests in 14 months, both in soil plus litter matrix from their ‘home’ forest and in those from the ‘away’ forest. We found that leaf (litter) trait variation among species and plant functional types, headed by species' dry matter content but also including tissue pH, was the strongest driver of variation in leaf litter decomposition rates. Surprisingly, however, while these effects of interspecific trait variation were very strong among species from the same site, there was no overall difference in litter decomposability between the species from the acidic versus calcareous site. Equally surprising was that this strong difference in pH of soil substrate plus litter matrix from an acidic sandstone site versus a calcareous karst site did not directly affect leaf litter decomposition rates across a given species set. This first attempt to disentangle the multiple potential direct and indirect ways in which soil and leaf (litter) acidity might be related to litter decomposition rates has important implications for our understanding of soil–plant feedbacks. Based on our forest‐based study, we predict that soil–plant feedbacks via acidity are unlikely to be strong in ecosystems with wide‐ranging species in terms of their leaf functional traits, including leaf pH. A free Plain Language Summary can be found within the Supporting Information of this article.

Seedling emergence and establishment are fragile processes that determine the direction and structure of forest succession and regeneration. However, seedling emergence and establishment are easily affected by biotic and abiotic (environmental) factors. A dense and expanding understory of dwarf bamboo is one such important factor that can seriously hinder the seedling regeneration. We conducted a field experiment to investigate the emergence and establishment of canopy tree seedlings under artificially controlled densities of dwarf bamboo. We found that understory dwarf bamboo obstructed seedling emergence but reduced the death of seedlings. Although understory dwarf bamboo reduced the median retention time of seedlings, dense bamboo increased the mean survival time of seedlings. Our results suggest that understory dwarf bamboo has multiple selectivities for tree seedling emergence and establishment: high‐density dwarf bamboo was beneficial to evergreen species but lower‐density of bamboo was conducive to the survival of deciduous species, it means the dwarf bamboo potentially alters successional trajectories of forest communities. Path analysis revealed that the most important factors affecting tree seedling emergence and death were the abundance of seeds in the seed bank and the density of emerged seedlings, and that the soil temperature promoted seedling emergence but increased seedling death, the thickness of litter limited seedling emergence, and the leaf area index of the bamboo canopy limited seedling death. The present study suggests that dwarf bamboo can directly alter the microenvironment, significantly reducing light levels and soil temperature but increasing the thickness of litter and soil humus, thereby indirectly impacting the regeneration of tree seedlings. Our results indicate that various factors affected seedling emergence, and there were complex indirect relationships among these factors. In general, biological factors had a stronger influence on tree seedling regeneration than environmental factors., Understory dwarf bamboo is expanding rapidly and resulting in irregular bamboo shrub layer formation in forests. This was not only can alter the ground surface environment (such as light, thickness of litter, thickness of soil humus, soil temperature, and soil humidity), but also slowed down the speed and density of seedling emergence, and accelerated the rate of seedling death; it has inhibited the establishment of local woody species, acting as an “ecological filter” in forest regeneration. And the multivariate path analysis showed that seed bank and soil temperature were the most important decision factors affecting seedling emergence, and the density of emerged seedling was the decision factor for seedling mortality, exerting a density dependence effect, while the litter thickness, leaf area index with dwarf bamboo canopy and dwarf bamboo density were the key limitation factors for seedling mortality, and soil temperature is the important promoting factor for seedling death.

Resource availability and heterogeneity are recognized as two essential environmental aspects to determine species diversity and community abundance. However, how soil resource availability and heterogeneity determine species diversity and community abundance in highly heterogeneous and most fragile karst landscapes is largely unknown. We examined the effects of soil resource availability and heterogeneity on plant community composition and quantified their relative contribution by variation partitioning. Then, a structural equation model (SEM) was used to further disentangle the multiple direct and indirect effects of soil resource availability on plant community composition. Species diversity was significantly influenced by the soil resource availability in shrubland and woodland but not by the heterogeneity in woodland. Abundance was significantly affected by both soil resource availability and heterogeneity, whereas variation partitioning results showed that soil resource availability explained the majority of the variance in abundance, and the contribution of soil resource heterogeneity was marginal. These results indicated that soil resource availability plays a more important role in determining karst plant community composition than soil resource heterogeneity. Our SEMs further found that the multiple direct and indirect processes of soil resource availability in determining karst species diversity and abundance were different in different vegetation types. Soil resource availability and heterogeneity both played a certain role in determining karst plant community composition, while the importance of soil resource availability far exceeded soil resource heterogeneity. We propose that steering community restoration and reconstruction should be highly dependent on soil resource availability, and multiple direct and indirect pathways of soil resource availability for structuring karst plant communities need to be taken into account., Both limited resource availability and significant resource heterogeneity are key features of karst landscapes. Our study first considered simultaneously resource availability and heterogeneity in karst regions and then quantified and compared the effects of resource availability and heterogeneity on community abundance and species diversity from the relevance‐theoretic perspective. More importantly, our study has important guiding significance to the vegetation restoration.

Background and Objectives: The landscape aesthetic service (LAS) is a crucial ecosystem service in subalpine forests of the mountain and ravine regions of the Tibetan Plateau, especially in autumn, it can provide important ecological and economic value, improving the livelihood of the local people. Canopy gap acts as a key role in the maintenance of species diversity and forest stability and ecosystem services. However, the mechanisms and interactions of forest stability maintained by gap formations and LAS via gap dynamics are not fully understood. The aim of this study was to explore the effects of canopy gaps on autumn color-leaved tree species diversity (ACTSD), color-leaved patch structure attributes (CPSA), LAS, and their interactions during the autumn viewing period, and to provide a fundamental basis for the management and protection of subalpine autumn color-leaved forests (SACF). Materials and Methods: We used very high-resolution images to quantify gap characteristics and examined the effects of canopy gaps on ACTSD, CPSA, and LAS in 21 SACF plots in southwestern China. We then used path analysis to determine the relationships between these factors. Results: The number of gaps and total gap area were the key gap characteristics affecting LAS, particularly, medium canopy gaps (51–100 m2) played a more important role. The path model showed that increasing the total gap area along with the number of medium canopy gaps had direct positive effects on ACTSD, color-leaved patch diversity, and total color-leaved patch area, thus improving the LAS. Conclusions: Canopy gaps indirectly improve LAS in autumn by significantly affecting ACTSD and CPSA. Our results suggest that forest managers may be able to manipulate the numbers and proportion of medium canopy gaps to protect and preserve ACTSD and color-leaved landscapes, promoting the LAS of SACF, and in turn, ensuring the coordinated development of economic, social, and ecological benefits for the underdeveloped rural montane areas of southwestern China.

BackgroundLandscape aesthetic service is a crucial ecosystem service in subalpine forests of the mountain and ravine regions of the Tibetan Plateau; it can provide important ecological and economic value, improving the livelihood of the local people. Canopy gap is recognized as the key mechanism in the maintenance of species diversity and forest stability and is known to further affect ecosystem service functioning. However, the mechanisms and interactions of forest stability maintained by gap formations and forest aesthetic service functioning via gap dynamics are not fully understood. We used very high-resolution images to quantify gap characteristics and examined the effects of canopy gaps on autumn colour-leaved tree species diversity, colour-leaved patch structure characteristics, and landscape aesthetic service in 21 subalpine autumn colour-leaved forest plots in southwestern China. We then used path analysis to determine the relationships between these factors.ResultsThe number of gaps and total gap area were the key gap characteristics affecting landscape aesthetic service; particularly, medium canopy gaps (51–100 m2) played a more important role. The path model showed that increasing the total gap area along with the number of medium canopy gaps had direct positive effects on autumn colour-leaved tree species diversity, colour-leaved patch diversity, and total colour-leaved patch area, thus improving the landscape aesthetic service.ConclusionCanopy gaps indirectly improve landscape aesthetic service by significantly affecting colour-leaved species diversity, colour-leaved patch diversity, and colour-leaved patch area. Our results suggest that forest managers may be able to manipulate the numbers and proportion of medium canopy gaps to protect and preserve autumn colour-leaved tree species diversity and colour-leaved landscapes, promoting the landscape aesthetic service of subalpine autumn colour-leaved forests and, in turn, ensuring the coordinated development of economic, social, and ecological benefits for the underdeveloped rural montane areas of southwestern China.

Background There has been considerable confusion concerning the number and classification of Sarcocystis spp. in chickens. Scarce nucleotide data of Sarcocystis spp. from chickens are provided in GenBank. The study aimed to investigate the morphological and molecular characteristics of Sarcocystis spp. found in chickens in China. Methods Tissues from 33 chickens were collected in 2019. Sarcocysts were observed using light (LM) and transmission electron microscopy (TEM). Individual sarcocysts from different chickens were selected for DNA extraction, and five loci, 18S rDNA, 28S rDNA, ITS1 region, the mitochondrial cox1 gene and the apicoplastic rpoB gene, were amplified from each sarcocyst, sequenced and analyzed. Results Only S. wenzeli was found in 14 of 33 (42.4%) chickens. Under LM, the sarcocysts were microscopic and exhibited palisade-like villar protrusions measuring 1.5–2.8 μm. Ultrastructurally, the sarcocyst wall contained numerous stubby hill-like villar protrusions. The protrusions included scattered microtubules, which extended from the tips of the protrusions into the ground substance. The five loci were successfully sequenced and the sequences deposited in GenBank. At 18S rDNA, ITS1 and cox1, the most similar sequences in GenBank were those of Sarcocystis sp. obtained from the brains of chickens, i.e. 99.9–100%, 98.1–98.5% and 99.3% identity, respectively. The five loci (18S rDNA, 28S rDNA, ITS1, cox1 and rpoB) showed different levels of interspecific sequence similarity with other closely related species of Sarcocystis (e.g. 99.8%, 99.0–99.2%, 89.3–89.7%, 98.5%, and 97.5%, respectively, with S. anasi). Phylogenetic analysis based on four of the loci (18S rDNA, cox1, rpoB and ITS1) revealed that S. wenzeli formed an independent clade with Sarcocystis spp. that utilize geese or ducks as intermediate hosts and canines as the known or presumed definitive host. Conclusions To our knowledge, the sequences of 28S rDNA and rpoB reported here constitute the first records of genetic markers of Sarcocystis spp. in chickens. Based on molecular analysis, S. wenzeli might be responsible for the neurological disease in chickens, and ITS1 and rpoB are more suitable for discriminating it from closely related Sarcocystis spp. Phylogenetic analysis revealed that S. wenzeli presents a close relationship with Sarcocystis spp. in geese or ducks.

Background: The effect of testosterone supplementation in patients with chronic heart failure (CHF) remains uncertain.Methods: A meta-analysis of randomized controlled trials (RCTs) was performed. RCTs that evaluate the chronic effect of testosterone supplementation on exercise capacity and cardiac function in CHF were identified via searching of PubMed, Embase, and the Cochrane's Library databases. Heterogeneity was evaluated by the Cochrane's Q test and I2 statistics. A fixed-effect model was used if the heterogeneity was not significant (I2 < 50%); otherwise, a random-effect model was applied.Results: Eight studies including 170 patients in the testosterone supplementation group and 162 in the control group were included. Overall, testosterone supplementation was not associated with an improved exercise capacity (walking test: standardized mean difference [SMD] = 0.36, p = 0.07). Sensitivity analyses limited to male patients showed similar results (SMD = 0.21, p = 0.15), and subgroup analyses also showed similar results in male HF patients with baseline total testosterone (TT) ≥ or < 10 nmol/L. However, patients with TT at endpoint ≥ 25 nmol/L was associated with improved exercise capacity (SMD = 1.12, p = 0.02), but not for those with TT at endpoint < 25 nmol/L (SMD = 0.24, p = 0.12). In addition, VO2max (weight mean difference [WMD] = 0.85, p = 0.43), the functional classification (the New York Heart Association classification: WMD = −0.08, p = 0.16) and quality of life (Minnesota Living with Heart Failure [MLHF] questionnaire: WMD = −6.03, p = 0.12) were not significantly affected. Moreover, testosterone supplementation did not significantly affect left ventricular ejection fraction (WMD: −1.52%, p = 0.37), serum B-type natriuretic peptide (SMD: −0.19, p = 0.23), or a composite outcome of death or HF hospitalization (risk ratio [RR]: 1.02, p = 0.96). Although testosterone supplementation increased systolic blood pressure (BP) in CHF patients (WMD: 5.68 mmHg, p < 0.001), diastolic BP or heart rate was not significantly changed as compared to control.Conclusions: Testosterone supplementation within a physiological range is not associated with significantly improved exercise capacity, cardiac function, quality of life, or clinical outcome in CHF patients.

We sought to explore factors associated with early pressure injury progression and build a model for predicting these outcomes using a machine learning approach.A retrospective cohort study.In this study, we recruited paediatric patients, with hospital-acquired stage I pressure injury or suspected deep tissue injury, who met the inclusion criteria between 1 January 2015-31 October 2018. We divided patients into two groups, namely healing or delayed healing, then followed them up for 7 days. We analysed patient pressure injury characteristics, demographics, treatment, clinical situation, vital signs, and blood test results, then build prediction models using the Random Forest and eXtreme Gradient Boosting approaches.The best prediction model, trained and tested using Random Forest with 10 variables, achieved an accuracy, sensitivity, specificity, and area under the curve of 0.82 (SD 0.06), 0.80 (SD 0.08), 0.84 (SD 0.08), and 0.89 (SD 0.06), respectively. The most contributing variables, in order of importance, included serum creatinine, red blood cell, and haematocrit.An awareness of specific conditions and areas that could lead to delayed healing pressure injury in paediatric patients is needed.This evidence-based prediction model, coupled with the aforementioned clinical indicators, is expected to enhance early prediction of outcomes in paediatric patients thereby improve the quality of care and the outcome of children with PIs.目的: 研究旨在探索与早期压力损伤进展相关的因素, 并利用机器学习方法建立结果预测模型。 设计: 回顾性列队研究。 方法: 本研究中, 成功招募2015年1月1日至2018年10月31日期间符合纳入标准的儿科患者, 此类患者经诊断为患有I期压疮或疑似患有深部组织损伤。我们将患者分为康复组和延迟康复组, 随访7天。研究过程中, 我们对患者的压力损伤特征、人口统计学特征、治疗、临床情况、生命体征和血液检测结果等进行分析,随后利用随机森林算法和极端梯度提升方法建立预测模型。 结果: 利用含10个变量的随机森林算法, 以便对最佳预测模型进行应用训练和检验, 其准确度、灵敏度、特异度和曲线下面积分别为0.82(SD 0.06) 、0.80(SD 0.08) 、0.84(SD 0.08) 和0.89(SD 0.06) 。根据重要性排序, 影响最大的变量包括血清肌酐、红细胞和红细胞比容。 结论: 应了解可致儿科患者压疮延迟愈合的具体情况和部位。 影响: 此预测模型以询证为基础,综合上述临床指标, 可增强儿科患者预后的早期预测, 提高治疗质量和压力损伤患儿的预后。.

Toxoplasma gondiiis an obligate intracellular parasite with worldwide distribution. Virulence ofT.gondiiis a multigenic trait. Genetic and virulence data forT.gondiiisolates from humans and animals in China have been reported. However, almost all biological materials used for genotyping ofT.gondiifrom humans and pigs were DNA samples prepared from tissues, andT. gondiistrains used for virulence analysis were isolated mainly from cats. In this study, one isolate from a dead human fetus was identified as type I (ToxoDB #10) while the two isolates from dead pigs were type Chinese I (ToxoDB #9) with PCR-restriction fragment length polymorphism using 10 markers (SAG1,SAG2,SAG3,BTUB,GRA6,c22–8,c29–2,L358,PK1and Apico). Three isolates were comfirmed as virulent strains in mice. By cloning and sequences analysis, all isolates contained aPvuII restriction site (572–577 bp) in the KHB fragment and five tandem repeats in the 5′ UTR region ofSAG1, which were associated withT.gondiivirulence. The type Chinese I isolates contained two deletions of 15 and 3 bp at positions 635 to 649 and 658 to 660 in theGRA6, which were correlated with genotype, but not with virulence. To our knowledge, this is the first report on the systematic analysis of murine virulence of type Chinese I strain from pigs, and the associations of sequences of the KHB fragment andSAG1with virulence of type Chinese I strain. The Chinese I genotype was more closely related to type II strains.

Toxoplasma gondii is an obligate intracellular parasitic protozoan with a worldwide distribution. The parasites in edible tissues of pigs and oocysts from cats are the major sources of T. gondii infection in humans. However, there are no data from sick pigs in veterinary clinics or from stray cats in Jiangsu Province, eastern China. In total, biological samples from 141 sick pigs and 64 stray cats were collected from this region. The rate of T. gondii infection in sick pigs was 46.81% using a polymerase chain reaction (PCR), and the overall prevalence of toxoplasmosis in stray cats was 34.38% by PCR and an enzyme-linked immunosorbent assay (ELISA). T. gondii was significantly more prevalent in lungs and heart than in liver and spleen (P 0.05). Age and geographic region were considered to be the main risk factors associated with T. gondii infection in these pigs. The DNA samples from 17 sick pigs and seven stray cats, were successfully genotyped by multilocus PCR-restriction fragment length polymorphism (PCR-RFLP) with 10 genetic markers [SAG1, SAG2 (5'-3'SAG2, alt. SAG2), SAG3, GRA6, PK1, c22-8, c29-2, BTUB, L358 and Apico]. Six distinct genotypes were found, which were designated ToxoDB PCR-RFLP genotypes #9 (Chinese I), #10 (Type I), #213, and #89, and New 1 and New 2. Chinese I is the most prevalent T. gondii genotype in this region. The two new genotypes (designated New 1 and New 2) are reported and the ToxoDB PCR-RFLP genotype #89 is found for the first time in China. Such information will be useful for the prevention, diagnosis and treatment of porcine toxoplasmosis in Jiangsu Province, eastern China.

Woody debris (WD) represents a globally significant carbon stock and its decomposition returns nutrients to the soil while providing habitat to microbes, plants and animals. Understanding what drives WD decomposition is therefore important. WD decomposition rates differ greatly among species. However, the role of bark in the process remains poorly known. We ask how, and how much, interspecific variation in bark functional traits related to growth and protection have afterlife effects on the decomposition of wood, partly mediated by animals. We examine the roles of bark cover and bark traits throughout the wood decomposition process. Synthesis. We find that: (1) bark effects on WD decomposition are species- and wood size-specific, (2) bark can enhance coarser WD decomposition but slows twig decomposition in some species, and (3) bark acts as an environmental filter to faunal assemblages in the early stage of wood decomposition. We highlight the need to account for bark effects on WD decomposition and offer an important complementary contribution to including woody species identity effects in biogeochemical and climate-change models via species bark traits.

Evasion strategies of intracellular parasites by hijacking cellular pathways, are necessary to ensure successful survival and replication. Eimeria tenella (E. tenella) has the ability to circumvent apoptosis of infected cells through increased expression of the transcriptional factor NF-κB and the anti-apoptotic factor Bcl-xL during the development of second generation schizonts. Artemisinin (ART) and its original plant, the dried leaves of Artemisia annua (LAA) have been shown to be effective against avian coccidiosis, however, the underlying mechanism remains unclear. We showed that E. tenella infection promoted the expression of anti-apoptotic protein Bcl-2 and inhibited the expression of pro-apoptotic proteins Bax and cleaved caspase-3 at 60 h post infection (PI), with a higher ratio of Bcl-2 to Bax. Nevertheless, the expression trends of Bcl-2, Bax and caspase-3 were reversed at 120 h and 192 h PI. ART treatment significantly abrogated Bcl-2 expression, whereas it promoted the expression levels of Bax and cleaved caspase-3 at the three time points above. Additionally, ART remarkably suppressed the increased mRNA expressions of NF-κB and interleukin-17A in ceca during infection by E. tenella. Compared with the ART treatment, LAA treatment exerted more improvements in clinical symptoms, promoting apoptosis and suppressing inflammatory response. These alterations caused by ART and LAA treatments were consistent with the reduced clinical diarrhea and pathological improvements in chicken ceca. Collectively, these results indicate that the inhibitory effects of ART or LAA on E. tenella infection may work through facilitating the apoptosis of infected host cells and inhibiting the inflammatory response.

Eimeria necatrix is one of the most pathogenic chicken coccidia and causes avian coccidiosis, an enteric disease of major economic importance worldwide. Eimeria parasites have complex developmental life cycles, with an exogenous phase in the environment and an endogenous phase in the chicken intestine. Oocysts excreted by chickens rapidly undergo meiosis and cell division to form eight haploid sporozoites (SZ). SZ liberated from sporocysts in the chicken intestine migrate to their preferred site of development to initiate cellular invasion. To date, almost nothing is known about the proteins that mediate parasite invasion in E. necatrix. In order to discover genes with functions involved in cellular invasion, the transcriptome profiles of E. necatrix unsporulated oocysts (UO) and SZ were analyzed using a combination of third-generation single-molecule real-time sequencing (TGS) and second-generation sequencing (SGS) followed by qRT-PCR validation. Correction of TGS long reads by SGS short reads resulted in 34,932 (UO) and 23,040 (SZ) consensus isoforms. After subsequent assembly, a total of 4949 and 4254 genes were identified from UO and SZ libraries, respectively. A total of 8376 genes were identified as differentially expressed genes (DEGs) between SZ and UO. Compared to UO, 4057 genes were upregulated and 4319 genes were downregulated in SZ. Approximately 1399 and 1758 genes were defined as stage-specific genes in SZ and UO, respectively. Gene Ontology (GO) classification and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that 2978 upregulated SZ genes were clustered into 29 GO terms, and 857 upregulated SZ genes were associated with 26 KEGG pathways. We also predicted a further 50 upregulated SZ genes and 73 upregulated UO genes encoding microneme proteins, apical membrane antigens, rhoptry neck proteins, rhoptry proteins, dense granule proteins, heat shock proteins, calcium-dependent protein kinases, cyclin-dependent kinases, cGMP-dependent protein kinase, and glycosylphosphatidylinositol-anchored surface antigens. Our data reveal new features of the E. necatrix transcriptional landscape and provide resources for the development of novel vaccine candidates against E. necatrix infection.

Properties of soil in karst regions are discontinuous and highly heterogeneous due to the adverse conditions of exposed rocks, tattered land form, steep slopes, and severe soil erosion. Uneven distribution of karst soil also leads to obvious spatial heterogeneity of moisture. Global precipitation changes might aggravate heterogeneity of soil moisture in soils with different thickness. Thus, exploring the responses of plants to water availability and soil heterogeneity in karst regions is necessary for the understanding of how precipitation changes might affect plant growth in soils with different thickness. Herbaceous plants especially grasses in karst regions are most easily to be affected by soil heterogeneity and water availability, considering they mainly utilize water and nutrients from the surface soil through their fibrous root system. Therefore, two graminaceous perennial grasses, Lolium perenne L. and Festuca arundinacea Schreb. were chosen for the present study. In addition, these two species are often chosen as pioneer plants for ecological restoration and reconstruction in karst regions because of their attributes of fast growth, strong adaptive ability, and high yield, which can effectively promote economic development and help to alleviate rural poverty in the harsh karst region. In our study, three water treatments (CK: 40 ml/day, D1: 20 ml/day and D2: 12 ml/day) were combined with three levels of soil thickness [shallow soil (SS; 5 cm), control (SCK; 15 cm) and deep soil (SD; 30 cm)] in a factorial randomized design and measurements were obtained of above- and below-ground growth, and biomass accumulation and allocation. The following results were obtained: (1) In CK water treatment, the total biomass, above-ground biomass, plant height and leaf area of both species were suppressed in SS as compared with those of SCK, and showed a decline to differing degrees, whereas these parameters were promoted in SD. The root biomass of L. perenne, and root length and surface area of both species in SS and SD were not significantly different to those of plants in SCK. The root biomass of F. arundinacea increased significantly in SS, but the observed values did not differ from those of the control (SCK). The specific root lengths of the two species decreased, and the ratio of root mass increased significantly in SS compared to SCK; but in SD, there was no difference compared to the control (SCK). (2) In D1 and D2 water treatments, there was a decrease in total biomass, above- and below-ground growth and biomass of both species in SS, and as water was reduced, the difference in plant height and leaf area between SS and SCK decreased in both species, and the difference in root length and root surface area between SS and SCK increased. In the SD treatment, apart from an increase in root length of F. arundinacea, there was no significant difference in other parameters of both species compared to SCK. The ratio of root mass of L. perenne in SS was still higher than SCK in D1 and D2 water treatments, but as water availability decreased from D1 to D2, the difference between SS and SCK decreased, and there was no significant difference between SD and SCK. There was no significant difference in the ratio root mass of F. arundinacea in both SS and SD compared to SCK under either D1 or D2 water treatments. The results of this study indicate that when water is sufficient, plant growth is restrained in shallow soil and promoted in deep soil, and as water decreases, plants in shallow and deep soil are both subjected to drought stress with resulting growth suppression, but the drought stress has a greater effect in shallow soil, and the drought conditions induced plant root depth increasing in deep soil. F. arundinacea, with a greater root depth, shows stronger adaptability to deep soil compared to L. perenne.

Due to the serious abrasion caused by the existence of solid phase in slurry, the slurry pumps usually adopt thick impeller blades to prolong its service life. In this paper, the influence of the blade thickness on the transient characteristics, the vibration, and the solid–liquid two-phase flow in a ceramic centrifugal pump with annular volute was investigated by the numerical method. And the experimental test was also conducted for computational fluid dynamics validation. The static pressure, the velocity, the force, and the solid fraction within the impeller passage and the volute casing have been discussed in detail. The results show that the influence of the blade thickness variation on the pressure fluctuation intensity is larger at the impeller inlet than that at the impeller exit and is larger at the pressure sides than that at the suction sides. The pressure fluctuation intensity decreases at the regions close to the volute tongue with the increase in the blade thickness. However, it increases at the regions, which are away from the volute tongue as the blade thickness increases. Moreover, the torque on the impeller, the radial force, and the axial force decreases with the increase in the blade thickness. Meanwhile, increasing the blade thickness will disperse the solid fraction at the impeller back shroud and make the regions with high solid fraction on pressure sides offset towards the impeller exit with a larger blade thickness, thus prolonging the service life of the impeller. However, the best efficiency point offsets to a low flow rate condition. The results are expected to pave the way for further optimization of impeller blade profile.

Background: Food safety incidents have aroused widespread public health concern, causing food price risk. However, the causal paths remain largely unexplored in previous literature. This paper sets out to identify the relations of local and spatial spillovers of food safety incidents and public health concerns to food price risk in consumer markets within a setting with heterogeneous food safety risk levels. Methods: (i) Theoretically, unlike prior work, this paper decomposes food safety risks into food safety incidents (objective incident component) and public health concern (subjective concern component). This article develops a theoretical framework of causality to capture the underlying causal pathways motivated by the theories of limited attention and two-step flow of communication. (ii) Empirically, using avian influenza shocks in China&rsquo, s poultry markets as natural experiments, this paper differentiates between low- and high-risk food and incidents. The article adopts dynamic spatial panel models to analyze potential nonlinearity, moderation, and mediation in the spillover of food safety risk to food price risk for a long panel of 30 provinces covering the November 2007 to November 2017 period. Results: (i) Food safety incident alone only triggers high-risk food price risk, not low-risk food price risk. (ii) Public health concern amplifies nonlinear food price risk triggered by food safety incident. (iii) High-risk incident intensifies negative pressure of public health concern on food price risk. (iv) Food safety incident indirectly affects high-risk food price risk through public health concern. Conclusions: Using a setting with heterogeneous risk levels, this paper documents that (i) food safety incident itself does not necessarily determine food price risk, whereas it is actually public health concern that directly causes nonlinear food price risk, (ii) public health concern spillover to food price risk is negatively moderated by high-risk incident, and (iii) food safety incident spillover to high-risk food price risk is mediated by public health concern. The findings complement current research by (i) elucidating the diverse impacts of food safety incident and public health concern on food price risk, which are obscure in previous literature, and (ii) highlighting that heterogeneous food and incident risk levels matter for determining food price risk spillover.

Gametocyte proteins of Eimeria spp. are essential components of the oocyst wall, and some of these proteins have been analysed to identify targets of transmission-blocking vaccines against avian coccidiosis. In the present study, a cDNA from E. necatrix gametocytes was cloned and sequenced. The cDNA is 1473 bp in length and encodes a 490-amino-acid protein containing a tyrosine-serine (Tyr/Ser)-rich domain and a proline-methionine (Pro/Met)-rich domain. A quantitative real-time PCR (qPCR) analysis showed that the cDNA is expressed only during gametogenesis. A fragment containing the Tyr/Ser-rich domain (rEnGAM59) was expressed in Escherichia coli BL21 (DE3) cells. Immunoblotting showed that rEnGAM59 was recognized by the serum of convalescent chickens after infection with E. necatrix, and that an anti-rEnGAM59 antibody recognized a ∼59 kDa protein and two other proteins (∼35 kDa and ∼33 kDa) in gametocyte extracts. An immunofluorescence assay showed that the anti-rEnGAM59 antibody recognized wall-forming bodies in the macrogametocytes and oocyst walls. An in vivo vaccination and challenge trial was conducted to test the potential utility of rEnGAM59 as a vaccine. Immunized chickens performed better than the unimmunized and challenged (positive control) chickens. The intestinal lesion scores were significantly lower in the immunized groups than in the positive control group (P0.05). In contrast, the body weight gains (BWG) were significantly higher in the immunized groups than in the positive control group (P0.05). There were no significant differences in the lesion scores and BWG between the groups immunized with rEnGAM59 protein or with live oocysts (P0.05). Chickens immunized with rEnGAM59 protein had a significantly higher antigen-specific serum IgY response (P0.05). rEnGAM59 protein can be used as candidate antigen to develop a recombinant coccidiosis vaccine.

Karst regions are characterized by heterogeneous soil habitats, with shallow wide soil (SW) on hilly slopes and deep narrow soil (DN) in rocky trenches. To make full use of limited water and nutrients, plants have therefore developed a number of root extension strategies. This study investigated the effect of soil moisture on horizontal root extension in SW and vertical root extension in DN by assessing root growth responses, biomass allocation, and root distribution. A full two-way factorial blocked design of soil dimensions by water availability was followed. The perennial grass Lolium perenne L. was grown in SW and DN under high (W100%), moderate (W50%), and low (W30%) water availability, respectively. The main results were as follows: (1) The total biomass of L. perenne was not influenced either by soil habitat or by water application. Root length, root surface area, root biomass and root to shoot ratio all decreased with decreasing water application in SW, but not in DN soil. (2) With decreasing water application, the cumulative percentage of root length, root surface area and root biomass in 4 rings from the center out to 12 cm of SW soil showed a trend of W50% > W30% > W100% in SW, however, the cumulative percentage of root biomass in 4 layers from the surface to a depth of 36 cm was not significantly different between different water treatments in DN. (3) Under all three water treatments, specific root length showed an increase but root length density showed a decreasing trend from the center outward in SW soil or from the surface to bottom in DN soil. Overall, these results suggest that in SW habitat, soil moisture determines horizontal expansion of the roots in L. perenne, although the overall expansion ability was limited in severe drought. However, due to the relatively strong water retention ability, soil moisture changes were less obvious in DN, resulting in no significant vertical extension of the root system. The root response of L. perenne helps our understanding of how herbaceous plants can adjust their belowground morphology to support their growth in harsh karst soil environments.

In this study, 36.8% (28/76) of tissue samples collected from domestic pigs (Sus scrofa) contained sarcocysts, as determined by light microscopy. The organisms were identified as Sarcocystis miescheriana and Sarcocystis suihominis based on their morphological and molecular characteristics. Four genetic markers, i.e., 18S rDNA, 28S rDNA, ITS-1 region (ITS-1), and the mitochondrial COX1 gene (COX1), of the two parasites were sequenced and analyzed, and the 28S rDNA and ITS-1 of S. suihominis obtained from pigs constituted the first records of these markers in GenBank. The sequences of the four loci (18S rDNA, 28S rDNA, ITS-1, and COX1) of S. miescheriana shared high identities with those of S. miescheriana obtained from domestic and/or wild pigs in GenBank, with similarities of 99.6%, 99.6%, 95.9%, and 95.4%, respectively. The 18S rDNA sequences of S. suihominis exhibited 99.4% identity with those of S. suihominis from domestic and wild pigs. The comparison of the newly obtained sequences of the four genetic markers between the two parasites revealed that the interspecific similarities of 18S rDNA, 28S rDNA, ITS-1, and COX1 were 97.7%, 96.6%, 80.3%, and 81.2%, respectively. Therefore, the two species could be better discriminated with ITS-1 and mitochondrial COX1 compared with 18S rDNA or 28S rDNA. The phylogenetic analysis using 28S rDNA indicated that the two Sarcocystis species in domestic pigs had a close relationship.

Animal disease is a major threat to the sustainability of the global livestock market. We explore the price risk spillover of avian influenza to the broiler market, from the perspective of public opinion. Unlike in previous work, where avian influenza is measured as a whole, we decompose an avian influenza epidemic into avian influenza outbreak and public opinion, measured by infection cases and Baidu and Google search volume. Theoretically, by introducing the theory of limited attention and two-step flow of communication, we develop an analytical framework to capture the causal mechanism of avian influenza outbreak, public opinion and broiler price risk spillover, arguing that it is actually public opinion, not avian influenza outbreak alone, that directly causes broiler price risk. Empirically, using a long panel from China spanning from November 2004-November 2017, we examine the causal mechanism and analyse the nonlinear spatial spillover of public opinion to broiler price risk. We find that: (i) neither poultry nor human infection with avian influenza outbreak has a significant spillover to broiler price, (ii) on average, public opinion has a negative spillover to broiler price, in general, spillover of public opinion to broiler price is inverse U-shaped, (iii) on average, public opinion has a negative direct effect on local broiler price and a three times larger negative spatial spillover effect on nearby broiler price, in general, direct and spatial spillover effects are inverse U-shaped. Our research highlights the importance of studying public opinion in amplifying price risk when analysing spillover of animal disease to the global livestock market.

Background Immunosuppression contributes to the mortality of sepsis. However, the underlying mechanism remains unclear. Methods In the present study, we investigated the role of inhibitory receptor immunoglobulin-like transcript 5 (ILT5) in sepsis. We first screened the expression of ILT family members, and we found that ILT5 was dramatically up-regulated in the peripheral blood mononuclear cells from sepsis patients versus healthy donors. Results Knockdown of ILT5 by small interfering ribonucleic acid increased bacterial killing and reactive oxygen species production in THP-1 and RAW264.7 cells. Moreover, ILT5-expressing monocytes/macrophages exhibited lower expression of antigen-presenting molecules including major histocompatibility complex-II and CD80. In the in vitro coculture system with monocytes/macrophages, blockage of ILT5 facilitated Th1 proliferation and differentiation of CD4+ T cells. Furthermore, in vivo experiments demonstrated that pretreatment with ILT5 blocking peptide improved the survival and pulmonary pathology of septic mice. Conclusions Together, our study identified ILT5 as an immunosuppressive regulator during sepsis, which may provide potential therapeutic strategy for sepsis.

Understory plant root proliferation into the litter layer occurs in many forest ecosystems and dramatically accelerates litter decomposition and nutrient release in the early stages. Mechanical fragmentation of leaf litter by fine roots could be one main reason, but its mechanism is poorly understood. To quantitatively assess the influence of fine roots on the mechanical fragmentation of leaf litter in the early decomposition process, we examined the effects of fine root mass of Lolium multiflorum on morphological and physical characteristics of decomposing litters of Symplocos setchuensis, a dominant species in a mid-subtropical evergreen broad-leaved forest of China, via a litter bag simulation experiment. The results showed that there were various fine root growth forms in/on decomposing litters, including the penetration and tearing of leaf epidermis, and the “walking or inserting” between the mesophyll tissue and the leaf veins. Fine root mass had a significant positive effect on the mass loss during litter decomposition (F = 8.543, P

As the critical component, the impellers of the slurry pumps usually have blades of a large thickness. The increasing excretion coefficient of the blades affects the flow in the impeller resulting in a relatively higher hydraulic loss, which is rarely reported. In order to investigate the influence of blade thickness on the transient flow characteristics of a centrifugal slurry pump with a semi-open impeller, transient numerical simulations were carried out on six impellers, of which the meridional blade thickness from the leading edge to trailing edge varied from 5-10 mm, 5-15 mm, 5-20 mm, 10-10 mm, 10-15 mm, and 10-20 mm, respectively. Then, two of the six impellers, namely cases 4 and 6, were manufactured and experimentally tested for hydraulic performance to verify the simulation results. Results of these tests agreed reasonably well with those of the numerical simulation. The results demonstrate that when blade thickness increases, pressure fluctuations at the outlet of the impeller become severe. Moreover, the standard deviation of the relative velocity in the middle portion of the suction sides of the blades decreases and that at the outlet of the impeller increases. Thus, the amplitude of the impeller head pulsation for each case increases. Meanwhile, the distribution of the time-averaged relative flow angle becomes less uniform and decreases at the outlet of the impeller. Hence, as the impeller blade thickness increases, the pump head drops rapidly and the maximum efficiency point is offset to a lower flow rate condition. As the thickness of blade trailing edge increases by 10 mm, the head of the pump drops by approximately 5 m, which is approximately 10 % of the original pump head. Futhermore, it is for the first time that the time-averaged relative flow angle is being considered for the analysis of transient flow in centrifugal pump. The presented work could be a useful guideline in engineering practice when designing a centrifugal slurry pump with thick impeller blades.

Twenty-one, 25-day-old, artificially reared, coccidia-free goslings (Anser cygnoides var. domestica) were inoculated orally with 0.5 × 104, 1 × 104, or 100 × 104 sporulated oocysts of Eimeria anseris and sacrificed at intervals from 24 to 216 h post-inoculation (HPI). Nine uninfected goslings served as negative controls. Parts of the visceral organs from goslings, including the intestines, kidneys, and liver, were fixed, sectioned, and observed microscopically. The results revealed that two generations of meronts occurred in the life cycle of E. anseris. The first generation of meronts developed at 24–96 HPI and the second generation at 90–128 HPI. Each meront contained 4–10 merozoites. Development of gamonts began at 128 HPI and mature oocysts appeared at 168 HPI. Developmental stages presented mainly in the epithelial cells of crypts and lamina propria in the posterior parts of the jejunum and ileum. Parasites localized mostly in the cytoplasm and occasionally in the nuclei of host cells. Histological lesions were pronounced in the jejunum and ileum. Desquamation and necrosis of the epithelium of intestine and crypts, infiltration of inflammatory cells, and hemorrhage and mucosal edema were associated with aggregates of endogenous stages. The infected goslings mainly showed severe diarrhea, depression, anorexia, and emaciation, suggesting that E. anseris is highly pathogenic in goslings.

Tree seed retention is thought to be an important factor in the process of forest community regeneration. Although dense understory dwarf bamboo has been considered to have serious negative effects on the regeneration of forest community species, little attention has been paid to the relationship between dwarf bamboo and seed retention. In a field experiment we manipulated the density of Fargesia decurvata, a common understory dwarf bamboo, to investigate the retention of seeds from five canopy tree species in an evergreen and deciduous broad-leaved mixed forest in Jinfoshan National Nature Reserve, SW China. We found that the median survival time and retention ratio of seeds increased with the increase in bamboo density. Fauna discriminately altered seed retention in bamboo groves of different densities. Arthropods reduced seed survival the most, and seeds removed decreased with increasing bamboo density. Birds removed or ate more seeds in groves of medium bamboo density and consumed fewer seeds in dense or sparse bamboo habitats. Rodents removed a greater number of large and highly profitable seeds in dense bamboo groves but more small and thin-husked seeds in sparse bamboo groves. Seed characteristics, including seed size, seed mass and seed profitability, were important factors affecting seed retention. The results suggested that dense understory dwarf bamboo not only increased seeds concealment and reduced the probability and speed of seed removal but also influenced the trade-off between predation and risk of animal predatory strategies, thereby impacting the quantity and composition of surviving seeds. Our results also indicated that dense understory dwarf bamboo and various seed characteristics can provide good opportunities for seed storage and seed germination and has a potential positive effect on canopy tree regeneration. (C) 2016 Elsevier Masson SAS. All rights reserved.

To quantitatively assess how fine root biomass and necromass of understorey species vary in the litter and upper mineral soil layers. The method combines in situ sampling of the litter layer with sequential coring during the root growth stage in three Chinese subtropical forests: coniferous forest, coniferous and broad-leaved mixed forest, and evergreen broad-leaved forest. Approximately 67–74 % of the total fine root mass occurred in the layer of decomposing litter (L2) and that of the top 7.5 cm of the soil (M1). Layer L2 was the preferential horizon for fine root growth, and the thickness of this horizon strongly affected the vertical fine root pattern. In addition, the fine root mass was positively correlated with total N concentration in layers L2 and M1. For all three forest types, most fine roots had diameters

Background: During zoonotic disease shocks (ZDSs), zoonotic disease outbreaks (ZDOs) can induce public health scares (PHSs), causing meat price risks (MPRs). Nevertheless, spatial spillovers of zoonotic disease shocks in meat markets remain unclear. We explore how zoonotic disease outbreaks and public health scares locally and spatially spill over to meat price risks, and whether spatial spillovers of public health scares decay with distance. Methods: (i) We construct a long panel covering 30 provinces and 121 months, using highly pathogenic avian influenza (HPAI) epidemics as exogenous shocks in Chinese meat sector. (ii) We decompose zoonotic disease shocks into zoonotic disease outbreaks (objective incident) and public health scares (subjective information) and examine their spillovers to meat price risks. (iii) We identify distance-decaying spatial spillovers of public health scares, by running our dynamic SAR models 147 times, from 80 km to 3000 km with 20 km as incremental value, in a setting with risk-level heterogeneity. Results: (i) Zoonotic disease outbreaks themselves only cause local and neighboring meat price risks for high-risk meat, not for low-risk or substitute meat. (ii) Public health scares exacerbate local and neighboring meat price risks for high-risk and low-risk meat, and local meat price risks for substitute meat. (iii) Spatial spillovers of public health scares are distance-decaying and U-shaped, with four spatial attenuation boundaries, and distance turning point is shorter for high-risk meat (500 km) than for low-risk meat (800 km). Conclusions: We complement the literature by arguing that health scares induced by disease outbreaks negatively spill over to meat prices, with U-shaped distance-decaying spatial effects. This suggests low interregional spatial market integration in meat products, due to distance decay of nonstandardized information and local government control effects, across provincial boundaries. To the best of our knowledge, we are the first to document nonmonotonic distance decay of health scare effects on food prices, previously not found by the literature.

Karst rocky outcrops divert water into the surrounding soil patches in a phenomenon known as the “funnel” effect. Soil depth is a key limiting factor in vegetation restoration in karst landscapes, yet few studies have examined the influence of the karst rock funnel effect on soil-limited plant community assemblages and productivity. We established artificial herbaceous communities with seeds from a karst species pool to evaluate how the species composition and productivity of karst herbaceous communities respond to soil depth and the funnel effect. Three months after species establishment, we found that all artificial communities were dominated by two grasses, Miscanthus sinensis and Lolium multiflorum. Plant abundance increased significantly with increasing soil depth, mainly owing to an increase in the individual numbers of the dominant species. The community coverage, photosynthetically active radiation (PAR) canopy capture rate, and shoot, root and total biomass also increased significantly with increasing soil depth. Funnel size and the soil depth × funnel size interaction significantly affected root biomass and root/shoot ratio; however, there was no effect of funnel size or the soil depth × funnel size interaction on species abundance, species richness and diversity, coverage, PAR canopy capture rate, shoot biomass, or total biomass. Therefore, this microcosm experiment provides evidence that the species composition and productivity of karst herbaceous communities are shaped by soil depth. The supply of water via the funnelling effect from karst rocky outcrops did not significantly affect herbaceous community assemblages or productivity, except in terms of root biomass. We suggest that the conservation and development of soil resources in environments with karst rocky outcrops may be more effective for the restoration of karst herbaceous communities than ensuring an adequate water supply.