Emerging fatal gout disease in Chinese goslings linked to acute kidney injury induced by novel goose astrovirus infection.

A novel goose astrovirus (GAstV) has broken out across China in recent years, causing widespread damage to the poultry industry. In goslings infected with GAstV, the leading cause of death is visceral gout. However, our understanding of the mechanism of gout formation in GAstV infection is largely inadequate. The aim of this study was to examine the pathogenicity of a GAstV strain and explore the molecular mechanisms of visceral gout caused by viral infection in goslings. The virulent GAstV strain HR2105/1 was effectively isolated from the visceral tissue of goslings in gout-affected areas. The whole genome of the HR2105/1 strain was sequenced and analyzed. Subsequently, we established a gosling gout models with experimental GAstV infection. Finally, we conducted a study on the mechanism of GAstV induced acute kidney injury. Phylogenetic analysis of the complete genome sequence showed that it was closely related to the strain circulating in China since 2016, and it was grouped within the GAstV-1 cluster. The clinical signs were reproduced by experimental infection of healthy goslings with the isolated strain and were found to be similar to those reported in clinical cases. Moreover, the virus exhibits strong renal tropism. Infection with the GAstV strain HR2105/1 was found to cause acute kidney injury, as evidenced by increased levels of uric acid and creatinine as well as severe pathological damage. Mechanistic experiments with Masson and Picrosirius Red staining revealed fibrosis in renal tissues after GAstV infection. Furthermore, TUNEL staining revealed that GAstV infection triggered renal cell apoptosis. Additionally, RT-qPCR revealed that GAstV infection caused an excessive inflammatory response by upregulating the expression of IL-1ß, IL-6, IL-10, TGF-ß, and iNOS in renal tissues. Overall, our findings demonstrate that GAstV infection causes renal damage by inducing renal cell apoptosis, fibrosis, and excessive inflammatory response, which subsequently leads to hyperuricemia and lethal visceral gout formation. This is the first systematic study on the etiology of lethal gout in goslings caused by GAstV infection, and we believe that the findings can guide vaccine development and therapeutic targets for GAstV-associated renal diseases.

Establishment of a reverse genetics system for virulent systemic feline calicivirus using circular polymerase extension reaction.

Feline caliciviruses can cause oral and upper respiratory tract infections in cats. However, a virulent and systemic feline calicivirus (VS-FCV) variant implicated in multisystem lesions and death in cats has emerged recently. To date, the mechanism underlying virulence variations in VS-FCV remains unclear. The aim of the present study was to provide a tool for exploring genetic variation in VS-FCV, by constructing an infectious clone of VS-FCV SH/2014. First, a full-length cDNA molecular clone of VS-FCV SH/2014 strain, which contains an Xba I recognition site generated by mutating one base (A→T) as a genetic marker, was constructed using the circular polymerase extension reaction (CPER) method. Second, the full-length cDNA clone was introduced into Crandell-Rees feline kidney cells using liposomes to rescue recombinant VS-FCV SH/2014 (rVS-FCV SH/2014). Third, the rescued viruses were identified by real-time PCR, immunofluorescence assay, western blotting, and electron microscopy. The full-length cDNA molecular clone of the VS-FCV SH/2014 strain was successfully constructed and that rVS-FCV SH/2014 could be rescued efficiently. rVS-FCV SH/2014 had the expected genetic markers and morphology and growth characteristics similar to those of the parental virus. The reverse genetics system provides a research platform for future studies on VS-FCV genetic variation and pathogenesis.

A getah virus-like-particle vaccine provides complete protection from viremia and arthritis in wild-type mice.

Getah virus (GETV) is an emerging mosquito-borne virus with economic impact on the livestock industry in East Asia. In this study, we successfully produced GETV virus-like particles (VLPs) in insect cells using the baculovirus expression vector system. We show that the GETV envelope glycoproteins were successfully expressed at the surface of the insect cell and were glycosylated. VLPs were isolated from the culture fluid as enveloped particles of 60-80 nm in diameter. Two 1 µg vaccinations with this GETV VLP vaccine, without adjuvant, generated neutralizing antibody responses and protected wild-type C57/BL6 mice against GETV viremia and arthritic disease. The GETV VLP vaccine may find application as a horse and/or pig vaccine in the future.

Machine learning-aided unveiling the relationship between chemical pretreatment and methane production of lignocellulosic waste.

Chemical pretreatment is a common method to enhance the cumulative methane yield (CMY) of lignocellulosic waste (LW) but its effectiveness is subject to various factors, and accurate estimation of methane production of pretreated LW remains a challenge. Here, based on 254 LW samples, a machine learning (ML) model to predict the methane production performance of pretreated feedstock was constructed using two automated ML platforms (tree-based pipeline optimization tool and neural network intelligence). Furthermore, the interactive effects of pretreatment conditions, feedstock properties, and digestion conditions on methane production of pretreated LW were studied through model interpretability analysis. The optimal ML model performed well on the validation set, and the digestion time, pretreatment agent, and lignin content (LC) were found to be key factors affecting the methane production of pretreated LW. If the LC in the raw LW was lower than 15%, the maximum CMY might be achieved using the NaOH, KOH, and alkaline hydrogen peroxide (AHP) with concentrations of 3.8%, 4.4%, and 4.5%, respectively. On the other hand, if LC was higher than 15%, only high concentrations of AHP exceeding 4% could significantly increase methane production. This study provides valuable guidance for optimizing pretreatment process, comparing different chemical pretreatment approaches, and regulating the operation of large-scale biogas plants.

Advances in microbial community, mechanisms and stimulation effects of direct interspecies electron transfer in anaerobic digestion.

Anaerobic digestion (AD) has been proven to be an effective green technology for producing biomethane while reducing environmental pollution. The interspecies electron transfer (IET) processes in AD are critical for acetogenesis and methanogenesis, and these IET processes are carried out via mediated interspecies electron transfer (MIET) and direct interspecies electron transfer (DIET). The latter has recently become a topic of significant interest, considering its potential to allow diffusion-free electron transfer during the AD process steps. To date, different multi-heme c-type cytochromes, electrically conductive pili (e-pili), and other relevant accessories during DIET between microorganisms of different natures have been reported. Additionally, several studies have been carried out on metagenomics and metatranscriptomics for better detection of DIET, the role of DIET's stimulation in alleviating stressed conditions, such as high organic loading rates (OLR) and low pH, and the stimulation mechanisms of DIET in mixed cultures and co-cultures by various conductive materials. Keeping in view this significant research progress, this study provides in-depth insights into the DIET-active microbial community, DIET mechanisms of different species, utilization of various approaches for stimulating DIET, characterization approaches for effectively detecting DIET, and potential future research directions. This study can help accelerate the field's research progress, enable a better understanding of DIET in complex microbial communities, and allow its utilization to alleviate various inhibitions in complex AD processes.

Regulation on C2-C8 carboxylic acid biosynthesis from anaerobic CO2 fermentation.

Bioconversion of CO2 into liquid fuels or chemicals, preferred medium chain carboxylic acids (caproic and caprylic acid), is an attractive CO2 utilization technology. The present study aims to investigate the effects of different ratios of H2/CO2 on regulating the distribution of C2-C8 carboxylic acid products, while the headspace pressure of 1.5 bar was set to amplify the effect of different ratios. The H2/CO2 ratio of 4:1 was more suitable for preparing acetic acid, where the highest acetic acid yield was 17.5 g/L. And the H2/CO2 ratio of 2:1 showed excellent chain elongation ability with the highest n-caprylic yield of 2.4 g/L. Additionally, the actual H2/CO2 ratios of 4:1 reactors were higher than that in 2:1 may be course chain elongation often accompanied by H2 production. The 16S rRNA genes analysis shows that the genus Terrisporobacter and Coriobacteriales may be related to acetic acid production enriched in H2/CO2 ratio 4:1 reactors, and the genus Clostridium and Paenibacillaceae may associate with the chain elongation pathway were enriched in H2/CO2 ratio 2:1 reactors.

Activation of CTU2 expression by LXR promotes the development of hepatocellular carcinoma.

Cytosolic thiouridylase 2 (CTU2) is an enzyme modifying transfer RNAs post-transcriptionally, which has been implicated in breast cancer and melanoma development. And we found CTU2 participated in hepatocellular carcinoma (HCC) progression here. HepG2 cells as well as xenograft nude mice model were employed to investigate the role of CTU2 in HCC development in vitro and in vivo respectively. Further, we defined CTU2 as a Liver X receptor (LXR) targeted gene, with a typical LXR element in the CTU2 promoter. CTU2 expression was activated by LXR agonist and depressed by LXR knockout. Interestingly, we also found CTU2 took part in lipogenesis by directly enhancing the synthesis of lipogenic proteins, which provided a novel mechanism for LXR regulating lipid synthesis. Meanwhile, lipogenesis was active during cell proliferation, particularly in tumor cells. Reduction of CTU2 expression was related to reduced tumor burden and synergized anti-tumor effect of LXR ligands by inducing tumor cell apoptosis and inhibiting cell proliferation. Taken together, our study identified CTU2 as an LXR target gene. Inhibition of CTU2 expression could enhance the anti-tumor effect of LXR ligand in HCC, identifying CTU2 as a promising target for HCC treatment and providing a novel strategy for the application of LXR agonists in anti-tumor effect.

Enhanced direct gaseous CO2 fixation into higher bio-succinic acid production and selectivity.

Utilizing CO2 for bio-succinic acid production is an attractive approach to achieve carbon capture and recycling (CCR) with simultaneous production of a useful platform chemical. Actinobacillus succinogenes and Basfia succiniciproducens were selected and investigated as microbial catalysts. Firstly, the type and concentration of inorganic carbon concentration and glucose concentration were evaluated. 6 g C/L MgCO3 and 24 g C/L glucose were found to be the optimal basic operational conditions, with succinic acid production and carbon yield of over 30 g/L and over 40%, respectively. Then, for maximum gaseous CO2 fixation, carbonate was replaced with CO2 at different ratios. The "less carbonate more CO2" condition of the inorganic carbon source was set as carbonate: CO2 = 1:9 (based on the mass of carbon). This condition presented the highest availability of CO2 by well-balanced chemical reaction equilibrium and phase equilibrium, showing the best performance with regarding CO2 fixation (about 15 mg C/(L·hr)), with suppressed lactic acid accumulation. According to key enzymes analysis, the ratio of phosphoenolpyruvate carboxykinase to lactic dehydrogenase was enhanced at high ratios of gaseous CO2, which could promote glucose conversion through the succinic acid path. To further increase gaseous CO2 fixation and succinic acid production and selectivity, stepwise CO2 addition was evaluated. 50%-65% increase in inorganic carbon utilization was obtained coupled with 20%-30% increase in succinic acid selectivity. This was due to the promotion of the succinic acid branch of the glucose metabolism, while suppressing the pyruvate branch, along with the inhibition on the conversion from glucose to lactic acid.

An adenovirus-vectored vaccine based on the N protein of feline coronavirus elicit robust protective immune responses.

Feline coronavirus (FCoV) is an unsegmented, single-stranded RNA virus belonging to the Alphacoronavirus genus. It can cause fatal feline infectious peritonitis (FIP) in cats of any ages. Currently, there are no effective prevention and control measures to against FCoV. In this study, we developed a recombinant adenovirus vaccine, AD5-N, based on the nucleocapsid(N) protein of FCoV. The immunogenicity of AD5-N was evaluated through intramuscular immunization in 6-week-old Balb/c mice and 9-12 months old cats. Compared to the control group, AD5-N specifically induced a significant increase in IgG and SIgA levels in the vaccinated mice. Furthermore, AD5-N not only effectively promoted strong cellular immune responses in cats but also induced high levels of specific SIgA, effectively helping cats resist FCoV infection. Our findings suggest that adenovirus vector vaccines based on the N gene have the potential to become candidate vaccines for the prevention and control of FCoV infection.

Performance and mechanism of various microaerobic pretreatments on anaerobic digestion of tobacco straw.

Tobacco straw is an abundant biomass in China's agricultural ecosystems, and has high potential for methane production. However, the anaerobic digestion (AD) efficiency is limited by the recalcitrant lignocellulose structure of the tobacco straw. In this study, three microaerobic pretreatments were performed for the AD of tobacco straw to increase methane production. Among them, microbial pretreatment with biogas slurry at an oxygen concentration of 4 mL/g VS resulted in the highest methane production of 349.1 mL/g VS, increasing by 19.8 % than that of untreated. During this pretreatment, the relative abundances of Enterococcus and Clostridium sensu stricto 12, which are closely related to acetic acid production and cellulose degradation, were high, and these bacteria might have an important contribution to substrate hydrolysis and the methanogenesis efficiency of the AD process. This study advances the understanding of microaerobic pretreatment processes and provides technological guidance for the efficient utilization of tobacco straw.

Machine learning-based prediction of methane production from lignocellulosic wastes.

The biochemical methane potential test is a standard method to determine the biodegradability of lignocellulosic wastes (LWs) during anaerobic digestion (AD) with disadvantages of long experiment duration and high operating expense. This paper developed a machine learning model to predict the cumulative methane yield (CMY) using the data of 157 LWs regarding physicochemical characteristics, digestion condition and methane yield, with the coefficient of determination equal to 0.869. Model interpretability analyses underscored lignin content, organic loading, and nitrogen content as pivotal attributes for CMY prediction. For the feedstocks with a cellulose content exceeding about 50%, the CMY in the early AD stage would be relatively lower than those with low cellulose content, but prolonging digestion time could promote methane production. Besides, lignin content in feedstock surpassing 15% would significantly inhibit methane production. This work contributes to valuable guidance for feedstock selection and operation optimization for AD plants.

Enhancing acidification efficiency of vegetable wastes through heat shock pretreatment and initial pH regulation.

Anaerobic digestion (AD) technology is a practical approach to alleviate severe environmental issues caused by vegetable wastes (VWs). However, its primary product is methane-rich biogas converted from the precursors (mainly volatile fatty acids, VFAs) after long fermentation periods, making traditional AD projects of low economic profits. Intervening in the methanogenesis stage artificially to produce high value-added VFAs can shorten the reaction time of the AD process and significantly improve profits, posing a promising alternative for treating VWs. Given this, this study applied heat shock (HS) pretreatment to inoculum to prevent methane production during AD and systemically investigated the effects of HS pretreatment and initial pH regulation on VFA production from VWs. The results showed that appropriate HS pretreatment effectively inhibited methane generation but promoted VFA accumulation, and VFA production was further enhanced by adjusting the initial pH to 8.0 and 9.0. The highest total VFA concentration of 14,883 mg/L with a VFA yield of 496.1 mg/gVS, 26.98% higher than that of the untreated group, was achieved at an initial pH 8.0 with HS pretreatment of 80 °C for 1 h. Moreover, pH regulation influenced the metabolic pathway of VFA production from VWs during AD, as butyrate was the dominant product at an initial pH of 6.0, while the increased initial pH improved the acetate proportion.

Pijx confers broad-spectrum seedling and panicle blast resistance by promoting the degradation of ATP ß subunit and OsRbohC-mediated ROS burst in rice.

Rice blast, caused by the fungal pathogen Magnaporthe oryzae, is one of the most important diseases of rice. Utilization of blast-resistance genes is the most economical, effective, and environmentally friendly way to control the disease. However, genetic resources with broad-spectrum resistance (BSR) that is effective throughout the rice growth period are rare. In this work, using a genome-wide association study, we identify a new blast-resistance gene, Pijx, which encodes a typical CC-NBS-LRR protein. Pijx is derived from a wild rice species and confers BSR to M. oryzae at both the seedling and panicle stages. The functions of the resistant haplotypes of Pijx are confirmed by gene knockout and overexpression experiments. Mechanistically, the LRR domain in Pijx interacts with and promotes the degradation of the ATP synthase ß subunit (ATPb) via the 26S proteasome pathway. ATPb acts as a negative regulator of Pijx-mediated panicle blast resistance, and interacts with OsRbohC to promote its degradation. Consistently, loss of ATPb function causes an increase in NAPDH content and ROS burst. Remarkably, when Pijx is introgressed into two japonica rice varieties, the introgression lines show BSR and increased yields that are approximately 51.59% and 79.31% higher compared with those of their parents in a natural blast disease nursery. In addition, we generate PPLPijx Pigm and PPLPijx Piz-t pyramided lines and these lines also have higher BSR to panicle blast compared with Pigm- or Piz-t-containing rice plants. Collectively, this study demonstrates that Pijx not only confers BSR to M. oryzae but also maintains high and stable rice yield, providing new genetic resources and molecular targets for breeding rice varieties with broad-spectrum blast resistance.

Genetic characterization of duck hepatitis B viruses from Anhui Province, China.

Duck hepatitis B virus (DHBV) infection model was frequently used as the experimental model for human hepatitis B virus (HBV) research. In order to decipher the genetic characteristics of DHBVs from Anhui province of China, 120 duck liver tissue samples were collected and subjected to PCR screening, and 28 samples were detected as DHBV positive. Subsequently, five DHBV-positive samples were selected for genome-wide amplification and a comprehensive analysis. Comparative analysis of complete genome sequences using the MegAlign program showed that five strains of DHBVs shared 94.5-96.3% with each other and 93.2-98.7% with other reference strains in GenBank. The phylogenetic analysis showed that all five DHBV strains belonged to the evolutionary branch of "Chinese DHBV" isolates or DHBV-2. Importantly, three potential intra-genotypic recombination events, between strains AAU-6 and Guilin, strains AAU-1 and GD3, and strains AAU-6 and AAU-1, were respectively found using the RDP and SimPlot softwares and considered the first report in avihepadnaviruses. These results not only improve our understanding for molecular prevalence status of DHBV among ducks, but also provide a reference for recombination mechanism of HBV.

Multistage orbital hybridization induced by multisite exchange interactions in high-entropy perovskites for high oxygen evolution reaction.

The oxygen evolution reaction (OER) has garnered considerable attention because of its promising prospects in electrochemical energy conversion applications, but a significant challenge is faced by the insufficient understanding of sluggish OER kinetics. In fact, the intrinsic "acceptance-donation" process of electrons between active sites and reactants is responsible for improving OER activity. Herein, we suggest a multielement hybridization strategy to rematch spin electron occupation and energy splitting in high-entropy perovskites with multiple orbital coordination. In this concept, electronic hopping between t2g and eg orbitals among particular catalytic sites can be obviously enforced due to introducing more favorable energy levels from neighboring metal sites, which can demonstrate multistage orbital hybridization reaction activity. As a result, our proposed multistage-hybridized high-entropy perovskites display an impressive activity of 199.8 mA cm-2 as an overpotential of âˆ¼0.46 V, which is âˆ¼5.3 times that of pristine perovskite. Different from traditional catalyst designs, this study focuses on multistage orbital hybridization and electron exchange interactions through a multisite coordination mechanism to construct a fast reaction pathway. Our findings provide a new strategy for accelerating OER catalytic kinetics.

Study on the Protective Immunity Induced by Pseudotyped Baculovirus Expressing the E Protein of Tembusu Virus in Ducklings.

The Duck Tembusu virus (DTMUV), a pathogenic flavivirus, has been causing significant economic losses in the Chinese poultry industry since 2010. This virus can severely decrease egg production and inhibit the growth of laying ducks and ducklings. While many vaccines have been developed to prevent DTMUV infection, fresh outbreaks continue to occur, as few effective vaccines are available. The E glycoprotein of DTMUV is the primary target for inducing protective immunity in the natural host. Therefore, we conducted an investigation and successfully developed a recombinant baculovirus containing the DTMUV E gene. Ducklings were then vaccinated with the purified protein derived from this virus as a potential vaccine candidate. Our findings demonstrated that the E glycoprotein of DTMUV was highly expressed in Sf9 cells. The vaccination of ducklings with the recombinant baculovirus Bac-E resulted in the induction of strong humoral and cellular immune responses. Most significantly, we observed that the vaccine provided 100% protective immunity against lethal challenges with the DTMUV YY5 strain.

A comprehensive review of the production, adoption and sustained use of biomass pellets in Ghana.

Ghana's Renewable Energy Master Plan of 2019 includes the production and use of biomass pellets. However, pellets have neither been developed commercially nor included in Ghana's energy mix. This paper reviewed the prospect of production, adoption and sustained use of pellets in Ghana. Besides having abundant biomass resources, Ghana has high market demand and relevant policies for pellet development. The production of pellets can significantly replace traditional household biomass demand and improve environmental and health quality. However, the production and use of pellets are limited due to technical, financial, social and policy issues. Our estimates show that 3% of the annual national average household income will be spent on pellet demand for cooking, with the highest burden on rural households in Ghana. Practical measures are required since the cost of pellets and gasifier stoves may limit pellet adoption and use in Ghana. Based on study findings, it is recommended that the government of Ghana establishes a robust supply chain and provides infrastructure for pellet production and use. Existing renewable energy policies should be reviewed to remove ambiguities, attract investment, and build capacity in the renewable energy sector. Apart from raising public awareness of the benefits of pellets use, the government of Ghana should ensure that continuous and thorough impact assessments are undertaken to assess the implications of pellet production and use. This review will inform policymaking on achieving sustainable production, adoption and use of pellets and assess Ghana's contribution to achieving the United Nations' sustainable development goals.

Pathogenicity and immunogenicity of gI/gE/TK-gene-deleted Felid herpesvirus 1 variants in cats.

BACKGROUND: Felid herpesvirus 1 (FHV-1) is a major pathogenic agent of upper respiratory tract infections and eye damage in felines worldwide. Current FHV-1 vaccines offer limited protection of short duration, and therefore, do not reduce the development of clinical signs or the latency of FHV-1. METHODS: To address these shortcomings, we constructed FHV ∆gIgE-eGFP, FHV ∆TK mCherry, and FHV ∆gIgE/TK eGFP-mCherry deletion mutants (ΔgI/gE, ΔTK, and ΔgIgE/TK, respectively) using the clustered regularly interspaced palindromic repeats (CRISPR)/CRISP-associated protein 9 (Cas9) system (CRISPR/Cas9), which showed safety and immunogenicity in vitro. We evaluated the safety and efficacy of the deletion mutants administered with intranasal (IN) and IN + subcutaneous (SC) vaccination protocols. Cats in the vaccination group were vaccinated twice at a 4-week interval, and all cats were challenged with infection 3 weeks after the last vaccination. The cats were assessed for clinical signs, nasal shedding, and virus-neutralizing antibodies (VN), and with postmortem histological testing. RESULTS: Vaccination with the gI/gE-deleted and gI/gE/TK-deleted mutants was safe and resulted in significantly lower clinical disease scores, fewer pathological changes, and less nasal virus shedding after infection. All three mutants induced virus-neutralizing antibodies after immunization. CONCLUSIONS: In conclusion, this study demonstrates the advantages of FHV-1 deletion mutants in preventing FHV-1 infection in cats.