H5N1 high pathogenicity avian influenza virus in migratory birds exhibiting low pathogenicity in mallards increases its risk of transmission and spread in poultry.

In 2020, an H5N1 avian influenza virus of clade 2.3.4.4b was detected in Europe for the first time and was spread throughout the world by wild migratory birds, resulting in the culling of an unprecedented number of wild birds and poultry due to the epidemic. In February 2023, we isolated and identified a strain of H5N1 high pathogenicity avian influenza virus from a swab sample from a grey crane in Ningxia, China. Phylogenetic analysis of the Hemagglutinin (HA) gene showed that the virus belonged to clade 2.3.4.4b, and several gene segments were closely related to H5N1 viruses infecting humans in China. Analysis of key amino acid sites revealed that the virus contained multiple amino acid substitutions that facilitate enhanced viral replication and mammalian pathogenicity. The results of animal challenge experiments showed that the virus is highly pathogenic to chickens, moderately pathogenic to BALB/c mice, and highly infectious but not lethal to mallards. Moreover, the virus exhibited minor antigenic drift compared with the H5-Re14 vaccine strain. To this end, we need to pay more attention to the monitoring of wild birds to prevent further spread of viruses to poultry and mammals, including humans.

Building-Envelope-Inspired, Thermomechanically Robust All-Fiber Ceramic Meta-Aerogel for Temperature-Controlled Dominant Infrared Camouflage.

The unsatisfactory properties of ceramic aerogels when subjected to thermal shock, such as strength degradation and structural collapse, render them unsuitable for use at large thermal gradients or prolonged exposure to extreme temperatures. Here, a building-envelope-inspired design for fabricating a thermomechanically robust all-fiber ceramic meta-aerogel with interlocked fibrous interfaces and an interwoven cellular structure in the orthogonal directions is presented, which is achieved through a two-stage physical and chemical process. Inspired by the reinforced concrete building envelope, a solid foundation composed of fibrous frames is constructed and enhanced through supramolecular in situ self-assembly to achieve high compressibility, retaining over 90% of maximum stress under a considerable compressive strain of 50% for 10 000 cycles, and showing temperature-invariance when compressed at 60% strain within the range of -100 to 500 °C. As a result of its distinct response to oscillation tolerance coupled with elastic recovery, the all-fiber ceramic meta-aerogel exhibits exceptional suitability for thermal shock resistance and infrared camouflage performance in cold (-196 °C) and hot (1300 °C) fields. This study provides an opportunity for developing ceramic aerogels for effective thermal management under extreme conditions.

Epidemiological investigation and pathogenicity analysis of waterfowl astroviruses in some areas of China.

Waterfowl astroviruses are mainly duck astroviruses and goose astroviruses, of which duck astroviruses (DAstV-3, -4), goose astroviruses (GoAstV-1, -2) are the four new waterfowl 21 astroviruses in recent years, which can lead to enteritis, viral hepatitis, gout and reduce the growth performance of waterfowl, affecting the healthy development of the waterfowl farming industry. Since no targeted drugs or vaccines on the market, studies on the epidemiology of the virus are necessary for vaccine development. In this study, we collected 1546 waterfowl samples from 13 provinces in China for epidemiological investigation. The results showed that 260 samples (16.8%) were positive. Four species of astrovirus were detected in 13 provinces except Fujian province. Among the four sites tested, the highest positive rates were found in farms and slaughterhouses. Cross-host and mixed infection were observed in four species of waterfowl astroviruses. The whole genome of 17 isolates was sequenced and compared with published sequences. Genetic evolution and homology analysis showed that the isolated strains had high similarity to their reference sequences. To assess the pathogenicity of GoAstV, 7-day-old goslings were inoculated with GoAstV-1 and GoAstV-2 by the intramuscular route, and infected geese showed similar clinical signs, such as anorexia, depression, and weight loss. Organ damage was seen after infection, with histopathological changes in the heart, liver, spleen, kidney, and intestine, and higher viral loads in throat and anal swabs. These findings increase our understanding of the pathogenicity of GoAstV-1 and GoAstV-2 in goslings and provide more references for future research.

Multiscale Synergetic Bandgap/Structure Engineering in Semiconductor Nanofibrous Aerogels for Enhanced Solar Evaporation.

Solar-driven interface evaporation has been identified as a sustainable seawater desalination and water purification technology. Nonetheless, the evaporation performance is still restricted by salt deposition and heat loss owing to weak solar spectrum absorption, tortuous channels, and limited plane area of conventional photothermal material. Herein, the semiconductor nanofibrous aerogels with a narrow bandgap, vertically aligned channels, and a conical architecture are constructed by the multiscale synergetic engineering strategy, encompassing bandgap engineering at the atomic scale and structure engineering at the nano-micro scale. As a proof-of-concept demonstration, a Co-doped MoS2 nanofibrous aerogel is synthesized, which exhibits the entire solar absorption, superhydrophilic, and excellent thermal insulation, achieving a net evaporation rate of 1.62 kg m-2 h-1 under 1 sun irradiation, as well as a synergistically efficient dye ion adsorption function. This work opens up new possibilities for the development of solar evaporators for practical applications in clean water production.

Rapid detection of H5 subtype avian influenza virus using CRISPR Cas13a based-lateral flow dipstick.

Due to its high mortality rate, highly pathogenic avian influenza (HPAI), a notifiable animal illness designated by the World Organisation for Animal Health (WOAH), has caused enormous financial losses to the poultry sector. The H5 subtype of avian influenza virus (H5-AIV) is regarded as the most common highly pathogenic avian influenza virus (HPAIV) that threatens public health and safety. Virus isolation and reverse transcription quantitative PCR (RT-qPCR) are usually used to detect H5-AIV and are important for the timely diagnosis and control of H5-AIV. However, these methods are time-consuming and require a significant amount of effort. In this study, we established a recombinase-aided amplification (RAA) combined with CRISPR-Cas13a and lateral flow dipstick (LFD) assay for the detection of H5-AIV. The results showed that the process can be completed within 40 min at 37°C. The method had a detection limit of 0.1 copy/µL, which was comparable to the RT-qPCR. There was no cross-reactivity with H3-AIV, H7-AIV, H9-AIV, H10-AIV, IBV, NDV, RVA and DAstV. The kappa value of RT-RAA-Cas13a-LFD and RT-qPCR in 380 clinical samples was 0.89 (κ>0.75). In conclusion, we established a convenient, efficient and accurate method to detect H5-AIV, and the results can be visualized and interpreted using LFD, which can be adapted to the needs of grassroots laboratories and field-deployable assays. This approach provides a new perspective for clinical H5-AIV diagnosis and has great potential for application in clinical quarantine of the poultry farming.

Development of a recombinase-aided amplification combined with a lateral flow dipstick assay for rapid detection of H7 subtype avian influenza virus.

Avian influenza viruses (AIV) pose a significant persistent threat to the public health and safety. It is estimated that there have been over 100 outbreaks caused by various H7 subtypes of avian influenza viruses (AIV-H7) worldwide, resulting in over 33 million deaths of poultry. In this study, we developed a recombinase-aided amplification combined with a lateral flow dipstick assay for the detection of hemagglutinin (HA) genes to provide technical support for rapid clinical detection of AIV-H7. The results showed that the assay can complete the reaction within 30 min at a temperature of 39°C. Specificity tests demonstrated that there was no cross-reactivity with other common poultry pathogens, including Newcastle disease virus (NDV) and infections bronchitis virus (IBV). The detection limit of this assay was 1 × 101 copies/µL, while RT-qPCR method was 1 × 101 copies/µL, and RT-PCR was 1 × 102 copies/µL. The κ value of the RT-RAA-LFD and RT-PCR assay in 132 avian clinical samples was 0.9169 (p < 0.001). These results indicated that the developed RT-RAA-LFD assay had good specificity, sensitivity, stability and repeatability and may be used for rapid detection of AIV-H7 in clinical diagnosis.

Correction: Establishment and application of a quadruple real-time RT-PCR for detecting avian metapneumovirus.

[This corrects the article DOI: 10.1371/journal.pone.0270708.].

Establishment of two assays based on reverse transcription recombinase-aided amplification technology for rapid detection of H5 subtype avian influenza virus.

IMPORTANCE: Avian influenza virus (AIV) subtype H5 is a highly contagious zoonotic disease and a serious threat to the farming industry and public health. Traditional detection methods, including virus isolation and real-time PCR, require tertiary biological laboratories and are time-consuming and complex to perform, making it difficult to rapidly diagnose H5 subtype avian influenza viruses. In this study, we successfully developed two methods, namely, RF-RT-RAA and RT-RAA-LFD, for rapid detection of H5-AIV. The assays are characterized by their high specificity, sensitivity, and user-friendliness. Moreover, the results of the reaction can be visually assessed, which are suitable for both laboratory testing and grassroots farm screening for H5-AIV.

Scalable Synthesis of Flexible Single-Atom Monolithic Catalysts for High-Efficiency, Durable CO Oxidation at Low Temperature.

The creation of single-atom catalysts in a large-size, high-yield, and stable form represents an important direction for high-efficiency industrial catalysis in the future. Herein, we report a strategy to synthesize flexible single-atom monolithic catalysts (SAMCs) based on the hierarchical 3D assembly of single-atom-loaded oxide ceramic nanofibers. The nanofibers, which can be produced in a continuous and scalable manner, serve as an ideal support for single atoms spontaneously and almost completely exposed at the surface through the Kirkendall effect-enabled in situ ion migration during the spinning process, resulting in both high yield and large loading quantity. Moreover, the hierarchical 3D assembly of these nanofibers into a porous, flexible structure endows the SAMCs with the advantages of sufficient infiltration and oscillation tolerance when faced with high-throughput gaseous media, leading to both high catalytic efficiency and excellent durability. As a proof-of-concept demonstration, a Pt SAMC is synthesized, which exhibits 100% CO oxidation at low temperature (∼170 °C), excellent invariance toward high-frequency (10 Hz) oscillation, and high structural stability from 25 to 300 °C. This work is beneficial for the large-scale production of SAMCs in broad industrial applications.

Inhibited Grain Growth Through Phase Transition Modulation Enables Excellent Mechanical Properties in Oxide Ceramic Nanofibers up to 1700 °C.

Oxide ceramics are widely used as thermal protection materials due to their excellent structural properties and earth abundance. However, in extremely high-temperature environments (above 1500 °C), the explosive growth of grain size causes irreversible damage to the microstructure of oxide ceramics, thus exhibiting poor thermomechanical stability. This problem, which may lead to catastrophic accidents, remains a great challenge for oxide ceramic materials. Here, a novel strategy of phase transition modulation is proposed to control the grain growth at high temperatures in oxide ceramic nanofibers, realizing effective regulation of the crystalline forms as well as the size uniformity of primary grains, and thus suppressing the malignant growth of the grains. The resulting oxide ceramic nanofibers have excellent mechanical strength and flexibility, delivering an average tensile strength as high as 1.02 GPa after being exposed to 1700 °C for 30 min, and can withstand thousands of flexural cycles without obvious damage. This work may provide new insight into the development of advanced oxide ceramic materials that can serve in extremely high-temperature environments with long-term durability.

The genetic and biological characterization of the first avian paramyxovirus serotype 14 isolated from chicken in China.

In October 2020, an avian paramyxovirus serotype 14 (APMV-14)-designated chicken/Fujian/2160/2020 (FJ2160) was isolated from tracheal and cloacal swab sample of chicken collected from live bird market in Fujian province in China during the active surveillance program. The complete genome of FJ2160 comprised 15,444 nucleotides (nt) complying with the paramyxovirus "rule of six" and encoded six non-overlapping structural proteins in the order of 3'-NP-P-M-F-HN-L-'5. The complete genome sequence analysis showed that FJ2160 had the highest identity (90.0%) with the APMV-14 isolated from Japan, while the nucleotide sequence identities of FJ2160 and other APMVs ranged from 42.4 to 51.1%. The F protein cleavage site was TREGR↓L, which resembled a lentogenic strain of APMV-1. Phylogenetic analysis revealed that the FJ2160 closest relative was APMV-14. The intracerebral pathogenicity index (ICPI) tests indicated that the virus was lentogenic. This is the first report of APMV-14 in China. These results provide evidence that APMV-14 could infect chickens and reveal the genetic characteristics and biological properties of the virus, which can help to better understand this new emerging APMV-14.

Hemagglutinin expressed by yeast reshapes immune microenvironment and gut microbiota to trigger diverse anti-infection response in infected birds.

Introduction: The H5N8 influenza virus is a highly pathogenic pathogen for poultry and human. Vaccination is the most effective method to control the spread of the virus right now. The traditional inactivated vaccine, though well developed and used widely, is laborious during application and more interests are stimulated in developing alternative approaches. Methods: In this study, we developed three hemagglutinin (HA) gene-based yeast vaccine. In order to explore the protective efficacy of the vaccines, the gene expression level in the bursa of Fabricius and the structure of intestinal microflora in immunized animals were analyzed by RNA seq and 16SrRNA sequencing, and the regulatory mechanism of yeast vaccine was evaluated. Results: All of these vaccines elicited the humoral immunity, inhibited viral load in the chicken tissues, and provided partial protective efficacy due to the high dose of the H5N8 virus. Molecular mechanism studies suggested that, compared to the traditional inactivated vaccine, our engineered yeast vaccine reshaped the immune cell microenvironment in bursa of Fabricius to promote the defense and immune responses. Analysis of gut microbiota further suggested that oral administration of engineered ST1814G/H5HA yeast vaccine increased the diversity of gut microbiota and the increasement of Reuteri and Muciniphila might benefit the recovery from influenza virus infection. These results provide strong evidence for further clinical use of these engineered yeast vaccine in poultry.

Genome sequence analysis of H7N3 subtype avian influenza virus originated from wild birds and its potential infectivity in mice.

In 2021, an H7N3 avian influenza virus (AIV) was isolated from a mallard in Tianhewan Yellow River National Wetland Park, Ningxia Hui Autonomous Region, China. Sequences analysis showed that this strain received its genes from H7, H6, H5, H3, and H1 AIVs of domestic poultry and wild birds in Asia and Europe. It was mild pathogenicity in mice. These results suggest the importance of continued surveillance of the H7N3 virus to better understand the ecology and evolution of the AIVs in poultry and wild birds and the potential threat to humans.

Reverse transcription recombinase-aided amplification assay for H5 subtype avian influenza virus.

BACKGROUND: The H5 subtype avian influenza virus (AIV) has caused huge economic losses to the poultry industry and is a threat to human health. A rapid and simple test is needed to confirm infection in suspected cases during disease outbreaks. METHODS: In this study, we developed a reverse transcription recombinase-aided amplification (RT-RAA) assay for the detection of H5 subtype AIV. Assays were performed at a single temperature (39 °C), and the results were obtained within 20 min. RESULTS: The assay showed no cross-detection with Newcastle disease virus or infectious bronchitis virus. The analytical sensitivity was 103 RNA copies/µL at a 95% confidence interval according to probit regression analysis, with 100% specificity. Compared with published reverse transcription quantitative real-time polymerase chain reaction assays, the κ value of the RT-RAA assay in 420 avian clinical samples was 0.983 (p < 0.001). The sensitivity for avian clinical sample detection was 97.26% (95% CI, 89.56-99.52%), and the specificity was 100% (95% CI, 98.64-100%). CONCLUSIONS: These results indicated that our RT-RAA assay may be a valuable tool for detecting H5 subtype AIV.

A molecular, epidemiological and pathogenicity analysis of pigeon paramyxovirus type 1 viruses isolated from live bird markets in China in 2014-2021.

To expand our understanding of the epidemiology of pigeon paramyxovirus type 1 (PPMV-1) in China, risk-based active surveillance was undertaken with pigeon swabs collected from live bird markets in 2014-2021. Seventy-six PPMV-1 strains were isolated from 12 provinces (60%) of the 20 provinces surveyed, and the positive rates of PPMV-1 varied from 0.50% to 3.19% annually. The complete genomic sequences of 18 representative viruses were analyzed, revealing a genome of 15,192 nucleotides, with the gene order 3'-NP-P-M-F-HN-L-5'. All isolates contained the 112RRQKRF117 cleavage site in the fusion (F) protein, a characteristic generally associated with virulent Newcastle disease viruses (NDVs), and the intracerebral pathogenicity index values (1.05-1.41) of four isolates indicated their virulence. A challenge experiment also demonstrated that all four isolates are pathogenic to pigeons, with morbidity rates of 60-100% and mortality rates of 0-30%. A further analysis of the functional domains of the F and HN proteins revealed several mutations in the fusion peptide, signal peptide, neutralizing epitopes, heptad repeat region, and transmembrane domains, and the substitution of cysteine residue 25 (C25Y) and substitutions in the HRb region (V287I) of the F protein and the transmembrane domain (V45A) of the HN protein may play important roles in PPMV-1 virulence. In a phylogenetic analysis based on the complete sequences of the F gene, all eighteen isolates all clustered into sub-genotype VI.2.1.1.2.2 (VIb) in class II, and shared high nucleotide sequence identity, indicating that the PPMV-1 strains in sub-genotype VI.2.1.1.2.2 are the predominant PPMV-1 viruses in pigeons in China and that the variations in these viruses have been relatively stable over the past 8 years. This study identifies the genetic and pathogenicity characteristics of the PPMV-1 strains prevalent in China and extends our understanding of the prevalence of this virus in China.

Establishment and application of a quadruple real-time RT-PCR for detecting avian metapneumovirus.

In order to develop an appropriate method for high-throughput detection of avian metapneumovirus, a quadruple real-time reverse-transcription polymerase chain reaction assay was established with four pairs of specific primers and four specific probes based on the G or M gene of aMPV-A, aMPV-B, aMPV-C and aMPV-D. Its specificity and sensitivity were evaluated, and clinical samples were tested by the method. The results showed that all the four subgroups of avian metapneumovirus can be detected in the quadruple real-time RT-PCR assay simultaneously, with a detection limit of 100-1000 cRNA copies/reaction. The other common poultry viruses were negative. In the avian clinical sample detection, 39 out of 1920 clinical samples collected from 8 provinces were positive. Compared with published RT-PCR assays, the κ value of the quadruple real-time RT-PCR assay in 1920 avian clinical samples was 1.000 (P < 0.001). The established method could be used for the rapid detection of the four subgroups of avian metapneumovirus with high specificity and high sensitivity.

Comparative Antigenicity and Pathogenicity of Two Distinct Genotypes of Highly Pathogenic Avian Influenza Viruses (H5N8) From Wild Birds in China, 2020-2021.

To date, there have been three epidemic waves of H5N8 avian influenza worldwide. The current third epidemic wave began in October 2020 and has expanded to at least 46 countries. Active and passive surveillance were conducted to monitor H5N8 viruses from wild birds in China. Genetic analysis of 10 H5N8 viruses isolated from wild birds identified two different genotypes. Animal challenge experiments indicated that the H5N8 isolates are highly pathogenic in chickens, mildly pathogenic in ducks, while pathogenicity varied in BALB/c mice. Moreover, there were significant differences in antigenicity as compared to Re-11 vaccine strain and vaccinated chickens were not completely protected against challenge with the high dose of H5N8 virus. With the use of the new matched vaccine and increased poultry immune density, surveillance should be intensified to monitor the emergence of mutant strains and potential worldwide spread via wild birds.

Emerging Novel Reassortant Influenza A(H5N6) Viruses in Poultry and Humans, China, 2021.

A novel highly pathogenic avian influenza A(H5N6) clade 2.3.4.4b virus was isolated from a poultry market in China that a person with a confirmed case had visited. Most genes of the avian and human H5N6 isolates were closely related. The virus also exhibited distinct antigenicity to the Re-11 vaccine strain.