Cell entry of bovine respiratory syncytial virus through clathrin-mediated endocytosis is regulated by PI3K-Akt and Src-JNK pathways.

Bovine respiratory syncytial virus (BRSV) is an RNA virus with envelope that causes acute, febrile, and highly infectious respiratory diseases in cattle. However, the manner and mechanism of BRSV entry into cells remain unclear. In this study, we aimed to explore the entry manner of BRSV into MDBK cells and its regulatory mechanism. Our findings, based on virus titer, virus copies, western blot and IFA analysis, indicate that BRSV enters MDBK cells through endocytosis, relying on dynamin, specifically via clathrin-mediated endocytosis rather than caveolin-mediated endocytosis and micropinocytosis. We observed that the entered BRSV initially localizes in early endosomes and subsequently localizes in late endosomes. Additionally, our results of western blot, virus titer and virus copies demonstrate that BRSV entry through clathrin-mediated endocytosis is regulated by PI3K-Akt and Src-JNK signaling pathways. Overall, our study suggests that BRSV enters MDBK cells through clathrin-mediated endocytosis, entered BRSV is trafficked to late endosome via early endosome, BRSV entry through clathrin-mediated endocytosis is regulated by PI3K-Akt and Src-JNK signaling pathways.

Effect of pyrolytic temperatures on the 2,4-dichlorophenol adsorption performance of biochar derived from Populus nigra.

To investigate the correlation between the physicochemical properties of biochar and its adsorption performance for 2,4-dichlorophenol (2,4-DCP), Populus nigra was subjected to oxygen-limited pyrolysis at temperatures ranging from 300 to 600 ℃. The experimental results showed that as the pyrolysis temperature increased, the specific surface area and degree of graphitization of the resultant biochar increased, but the amount of oxygen-containing functional groups decreased. Populus nigra biochar produced at 450 ℃ exhibits the best adsorption performance for 2,4-DCP due to its excellent physicochemical properties and greater electron exchange capability. The removal of 2,4-DCP is a multi-step adsorption process dominated by chemisorption, which involved oxygen-containing functional groups-mediated hydrogen bonding, as well as π-π electron donor-acceptor (EDA) interaction between the aromatic rings and Cl atoms. The study highlights the potential of Populus nigra residues for producing biochar as an affordable and effective adsorbent for 2,4-DCP removal.

Insights into the enhanced effect of biochar on cadmium removal in vertical flow constructed wetlands.

Biochar has been increasingly applied in constructed wetlands (CWs) to remediate heavy metal (HM)-polluted water. Nevertheless, only few studies have elucidated the enhanced mechanism and potential synergies related to the HM removal from biochar-based CWs (BC-CWs) for HMs removal. This study used cadmium (Cd) as the target HM and added biochar into CWs to monitor physicochemical parameters, plant' physiological responses, substrate accumulation, and microbial metabolites and taxa. In comparison with the biochar-free CW (as CWC), a maximum Cd2+ removal of 99.7% was achieved in the BC-CWs, associated with stable physicochemical parameters. Biochar preferentially adsorbed the available Cd2+ and significantly accumulated Fe/Mn oxides-bond and the exchangeable Cd fraction. Moreover, biochar alleviated the lipid peroxidation (decreased by 36.4%) of plants, resulting in improved growth. In addition, extracellular polymeric substances were increased by 376.9-396.8 mg/L in BC-CWs than compared to CWC, and N and C cycling was enhanced through interspecific positive connectivity. In summary, this study explored comprehensively the performance and mechanism of BC-CWs in the treatment of Cd2+-polluted water, suggesting a promising approach to promote the plant-microbe-substrate synergies under HM toxicity.

Quantitative real-time PCR detection and analysis of a lumpy skin disease outbreak in Inner Mongolia Autonomous Region, China.

Lumpy skin disease (LSD) is a severe disease of bovine characterized by nodules on the skin, mucous membranes, and profuse nasal discharge which causes severe economic losses. In October 2020, an LSD outbreak case was found in Inner Mongolia Autonomous Region, China. A total of 1,206 cattle were sold from the same imported animal quarantine field to 36 farms after the quarantine period finished, and over 30 farmers reported symptoms such as skin scabs found in newly arrived cattle shortly after that. A large-scale LSD outbreak investigation was launched after laboratory diagnosis confirmed LSD. The clinical samples of 1,206 cattle from 36 farms, including 1,206 whole blood, 1,206 oral and nose swabs, and 355 scabs, were collected for the qRT-PCR test. The result showed that 51 whole blood samples (4.23%), 580 swab samples (48.09%), and 350 skin scabs (98.59%) were lumpy skin disease virus (LSDV) positive, 33 of 36 farms were affected. This study aims to provide a basis for LSD epidemiological traceability, movement control, and measures for prevention and control.

Enhanced removal of heavy metals and metalloids by constructed wetlands: A review of approaches and mechanisms.

Constructed wetlands (CWs) are increasingly employed to remediate heavy metal and metalloid (HMM)-polluted water. However, the disadvantages of HMM removal by conventional CWs (without enhancement), such as an unstable and unpredictable removal efficiency, hinder the reliability of this technology. The objective of this study was to review research on enhanced CWs for HMM removal. In particular, we performed a bibliometric analysis to evaluate research trends, critical literature, and keyword evolution in recent years. Subsequently, we reviewed various enhanced approaches for the application of CWs for the removal of HMMs, including the use of improved substrates, aquatic macrophytes, microorganisms, bioelectrochemical coupling systems, hybrid CW, external additives, and operation parameters. Furthermore, the main mechanisms underlying HMM removal by these approaches are summarized. Our review clearly reveals that research on the remediation of HMM-polluted water via CW technology is receiving increased attention, with no apparent trends in topics. The selection of appropriate enhanced approaches or operation parameters as well as methodological improvements should be based on the dominant environmental conditions of the CW column and removal mechanisms for the targeted HMMs. Based on the established literature, several suggestions are proposed to guide the optimization of the design and operation of efficient CWs for the treatment of HMM-polluted water.

Degradation of oxytetracycline by magnetic MOFs heterojunction photocatalyst with persulfate: high stability and wide range.

Photocatalysis with persulfate (PS) is an effective method for the degradation of degrading organic pollutants. In this study, Fe3O4/MIL-101(Fe), a magnetic heterojunction photocatalyst, was produced using a hydrothermal method. The material coupled with PS exhibited excellent removal efficiency for oxytetracycline (OTC) (87.1%, 1 h). And it has a wide range of applications, with good removal efficiency for OTC concentrations of 30 to 70 mg/L and pH values of 3 to 9. •SO4- and •OH played a major role in the OTC removal reaction and there was an Fe(III)/Fe(II) cycle during the reaction. With excellent stability and recoverability, the OTC removal efficiency decreased by only 4.29% after four cycles, and the Fe leaching did not exceed 0.035 mg/L per cycle. This study provides significant insights into the removal of organic pollutants from water bodies.

Reutilization of manganese enriched biochar derived from Phytolacca acinosa Roxb. residue after phytoremediation for lead and tetracycline removal.

This study aimed to evaluate the chemical form variation of Mn in Phytolacca acinosa Roxb. residue under different pyrolysis temperatures and its contribution to decontamination efficacy of lead (Pb(II)) and tetracycline (TC). The results illuminated that pyrolysis temperature is a crucial factor of fraction and bioavailability of Mn and other heavy metals in the resultant biochar and pyrolysis temperature under 450 °C may be most suitable for reutilization without potential risk. The Mn-enriched phytolaccaceae biochar (PSB450) exhibited more preferential sorption toward Pb(II) (279.33 mg/g) and TC (47.51 mg/g) than pristine phytolaccaceae biochar in the single system, mainly due to the formation of MnOx and Mn minerals via pyrolysis. Binary adsorption showed that Pb(II) would serve as a bridge between PSB450 and TC by complexation within a limited concentration range, thus facilitating their joint decontamination. This study provided an efficient alternative approach for reutilization of Mn-contaminated biomass.

Stimulation of pyrolytic carbon materials as electron shuttles on the anaerobic transformation of recalcitrant organic pollutants: A review.

Pyrolytic carbon materials (PCMs) with various surface functionalities are widely used as environmentally friendly and cost-efficient adsorbents for the removal of organic and inorganic pollutants. Recent studies have illustrated that PCMs as electron shuttles (ESs) could also show excellent performances in promoting the anaerobic transformation of recalcitrant organic pollutants (ROPs). Numerous studies have demonstrated the excellent electron-shuttle capability (ESC) of PCMs to stimulate the anaerobic reductive transformation of ROPs. However, there is a lack of consistent understanding of the mechanism of ESC formation in PCMs and the stimulation mechanism for ROPs anaerobic transformation. To gain a more comprehensive understanding of the latest developments in the study of PCMs as ESs for ROPs anaerobic transformation, this review summarizes the formation mechanism, influencing factors, and stimulation mechanisms of ESC. ESC benefits from redox functional groups (quinone and phenol groups), persistent free radicals (PFRs), redox-active metal ions, conductive graphene phase, and porous nature of their surface. The factors influencing ESC include the highest treatment temperature (HTT), feedstocks, modification methods, and environmental conditions, of which, the HTT is the key factor. PCMs promote the reductive transformation of ROPs under anaerobic conditions via abiotic and biotic pathways. Eventually, the prospects for the ROPs anaerobic transformation enhanced by PCMs are proposed.

Clinical Validation of Two Recombinase-Based Isothermal Amplification Assays (RPA/RAA) for the Rapid Detection of African Swine Fever Virus.

African swine fever (ASF), caused by African swine fever virus (ASFV), is a devastating infectious disease of domestic pigs and wild boars, and has tremendous negative socioeconomic impact on the swine industry and food security worldwide. It is characterized as a notifiable disease by World Organisation for Animal Health (OIE). No effective vaccine or treatment against ASF has so far been available. Early detection and rapid diagnosis are of potential significance to control the spread of ASF. Recombinase-based isothermal amplification assay, recombinase polymerase amplification (RPA) developed by TwistDx (Cambridge, United Kingdom) or recombinase-aided amplification (RAA) by Qitian (Wuxi, China), is becoming a molecular tool for the rapid, specific, and cost-effective identification of multiple pathogens. In this study, we aim to investigate if RPA/RAA can be a potential candidate for on-site, rapid and primary detection of ASFV. A panel of 152 clinical samples previously well-characterized by OIE-recommended qPCR was enrolled in this study, including 20 weak positive (Ct value ≥ 30) samples. This panel was consisted of different types, such as EDTA-blood, spleen, lung, lymph node, kidney, tonsil, liver, brain. We evaluated two recombinase-based isothermal amplification assays, RPA or RAA, by targeting the ASFV B646L gene (p72), and validated the clinical performance in comparison with OIE real-time PCR. Our result showed that the analytical sensitivity of RPA and RAA was as 93.4 and 53.6 copies per reaction, respectively at 95% probability in 16 min, at 39°C. They were universally specific for all 24 genotypes of ASFV and no cross reaction to other pathogens including Classical swine fever virus (CSV), Foot-and-mouth disease virus (FMDV), Pseudorabies virus, Porcine circovirus 2 (PCV2), Porcine Reproductive and respiratory syndrome virus (PPRSV). The results on detection of various kinds of clinical samples indicated an excellent diagnostic agreement between RPA, RAA and OIE real-time PCR method, with the kappa value of 0.960 and 0.973, respectively. Compared to real-time PCR, the specificity of both RPA and RAA was 100% (94.40% ∼ 100%, 95% CI), while the sensitivity was 96.59% (90.36% ∼ 99.29%, 95% CI) and 97.73% (92.03% ∼ 99.72%, 95% CI), respectively. Our data demonstrate that the developed recombinase-based amplification assay (RPA/RAA), promisingly equipped with field-deployable instruments, offers a sensitive and specific platform for the rapid and reliable detection of ASFV, especially in the resource-limited settings for the purpose of screening and surveillance of ASF.

Enhanced Cd2+ and Zn2+ removal from heavy metal wastewater in constructed wetlands with resistant microorganisms.

The bioaugmentation role of microbes is often impeded by heavy metal (HM) ions in constructed wetlands (CWs). To explore the interaction between microbes and HM ions, two identical CWs: an MCW (with resistant microorganisms) and a CCW (as control) were used in this study. Experiments analyzed static adsorption performance in a synthetic HM solution. The removal performance of Cd2+ and Zn2+ was further investigated in both CWs. The removal efficiencies (REs) of 81.92-99.56% and 74.05-98.79% were achieved for Cd2+ and Zn2+ in the adsorption study, respectively. Significantly higher REs of Cd2+ (99.60%), and Zn2+ (94.41%) were achieved in the MCW. The microbial community analysis revealed that the dominant genera were Serratia and Pseudomonas in the MCW. The subcellular analysis further demonstrated that the HMs bioaccumulated mainly in the cytomembrane and cell wall. These results indicate that CW with resistant microorganisms inoculated was an effective strategy for treating HMs wastewater.

Genome comparison of African swine fever virus China/2018/AnhuiXCGQ strain and related European p72 Genotype II strains.

African swine fever was introduced into China in August 2018 and led to high mortality in domestic pigs. We reported the genome characterization of the China/2018/AnhuiXCGQ strain mainly based on next-generation sequencing and comparison with related European p72 Genotype II strains. The genome was 189,393 bp long, encoding 181 open reading frames. Pair-wise genome sequence comparison revealed 54-107 variation sites between China/2018/AnhuiXCGQ and the other genotype II virulent strains contributing to the change of expression or alteration of amino acid residues in 10-38 genes. China/2018/AnhuiXCGQ strain shared the highest similarity with POL/2015/Podlaskie strain. Phylogenetic analysis based on a 125 kb long conserved central region revealed that the China/2018/AnhuiXCGQ strain and four European genotype II strains were grouped into three clusters. This study expanded our knowledge on the genetic diversity and evolution of ASFV and provided valuable information for diagnosis improvement and vaccine development.

Small interfering RNAs targeting peste des petits ruminants virus M mRNA increase virus-mediated fusogenicity and inhibit viral replication in vitro.

Peste des petits ruminants (PPR), caused by peste des petits ruminants virus (PPRV), is an acute or subacute, highly contagious and economically important disease of small ruminants. The PPRV is classified into the genus Morbillivirus in the family Paramyxoviridae. The PPRV matrix (M) protein possesses an intrinsic ability to bind to lipid membranes, and plays a crucial role in viral assembly and further budding. In this study, three different small interfering RNAs (siRNA) were designed on the basis of translated region for PPRV Nigeria 75/1M mRNA, and were subsequently synthesized for their transfection into Vero-SLAM cells, followed by infection with PPRVs. The results showed that two out of three siRNAs robustly induced cell-to-cell fusion as early as 36h post-infection with PPRVs, effectively suppressed expression of the M protein by interference for the M mRNA, and eventually inhibited viral replication in vitro. These findings led us to speculate that siRNA-mediated knockdown of the M protein would alter its interaction with viral glycoproteins, thus exacerbating intercellular fusion but hampering virus release.

Allelic variants of PRNP in 16 Chinese local sheep breeds.

Here, polymorphisms of the ovine prion protein gene were analyzed in 486 Chinese sheep from 16 main local breeds. Polymorphisms R or H at codons 154 and four polymorphisms at codon 171 encoding Q, R, H, or K were identified. The A/V polymorphism at codon 136 was not observed, and all sheep were homozygous for A at this position. In addition, ten polymorphisms at codons 21, 101, 112, 127, 138, 141, 143, 146, 153 and 189 were detected. The predominant Q allele occurred at codon 171, with a high frequency of 88.68 %, implying a risk of scrapie in China.