Toxoplasma gondii(T.gondii)is an obligate intracellular parasite with a wide range of hosts,including humans and many warm-blooded animals.The parasite exists in two interconvertible forms,namely tachyzoites and brady...Toxoplasma gondii(T.gondii)is an obligate intracellular parasite with a wide range of hosts,including humans and many warm-blooded animals.The parasite exists in two interconvertible forms,namely tachyzoites and bradyzoites in intermediate hosts that are responsible for acute and chronic infections respectively.Mature bradyzoites accumulate large amounts of amylopectin granules but their roles have not been fully characterized.In this study,the predicted key enzymes involved in amylopectin synthesis(UDP-sugar pyrophospharylase,USP)and degradation(alpha-glucan water dikinase,GWD)of ME49 strain were individually knocked out,and then bradyzoite-related phenotyping experiments in vitro and in vivo were performed to dissect their roles during parasite growth and development.Deletion of the usp or gwd gene in the type II strain ME49 reduced the replication rates of tachyzoites in vitro and parasite virulence in vivo,suggesting that amylopectin metabolism is important for optimal tachyzoite growth.Interestingly,the Δusp mutant grew slightly faster than the parental strain under stress conditions that induced bradyzoite transition,which was likely due to the decreased efficiency of bradyzoite formation of theΔusp mutant.Although theΔgwd mutant could convert to bradyzoite robustly in vitro,it was significantly impaired in establishing chronic infection in vivo.Both the Δusp and Δgwd mutants showed a dramatic reduction in the reactivation of chronic infection in an in vitro model.Together,these results suggest that USP and GWD,which are involved in amylopectin synthesis and degradation have important roles in tachyzoite growth,as well as in the formation and reactivation of bradyzoites in T.gondii.展开更多
Advances in microbiology rely on innovations in technology. Droplet microfluidics, as a versatile and powerful technique that allows high-throughput generation and manipulation of subnanoliter volume droplets, has bec...Advances in microbiology rely on innovations in technology. Droplet microfluidics, as a versatile and powerful technique that allows high-throughput generation and manipulation of subnanoliter volume droplets, has become an indispensable tool shifting experimental paradigms in microbiology. Droplet microfluidics has opened new avenues to various microbiological research, from resolving single-cell heterogeneity to investigating spatiotemporal dynamics of microbial communities, from precise quantitation of microbiota to systematic decipherment of microbial interactions, and from isolating rare and uncultured microbes to improving genetic engineered strains. In this review, we present recent advances of droplet microfluidics in various fields of microbiology: i) microbial cultivation, ii) microorganism detection and characterization, iii) antibiotic susceptibility testing, iv) microbial interactions, v) microbial biotechnology.We also provide our perspectives on the challenges and future directions for droplet microfluidic-based microbiology research.展开更多
The development of alternative therapies to treat chicken coccidiosis has become a hot topic because of the widespread use of conventional medicines.This study aimed to investigate the effectiveness of eugenol in trea...The development of alternative therapies to treat chicken coccidiosis has become a hot topic because of the widespread use of conventional medicines.This study aimed to investigate the effectiveness of eugenol in treating Eimeria tenella infection in broilers.Broiers,at the age of 14 d,were orally infected with sporulated Eimeria tenella oocysts,and then,eugenol essential oil was added to chicken feed at three different dosages(0.1,0.2 or 0.4 g/kg).The anticoccidial effects of eugenol essential oil were assessed using the anticoccidial index(ACI).As a result,eugenol exhibited a moderate anticoccidial effect,with an ACI of 167.37 at 0.2 g/kg.After eugenol treatment,the expression of occludin in the epithelial cells of the chicken cecum was significantly greater(P<0.05)than that in the epithelial cells of the nontreated control(IC)group.The proportion of intestinal Lactobacillus_agilli increased.Eugenol therapy dramatically increased the activity of superoxide dismutase.After high-dose treatment,the expression of the proinflammatory factors IL-1βand IL-6 significantly decreased,while the expression of the cytokines IL-4 and IFN-γsignificantly increased.The safety of eugenol essential oil was evaluated at the 1,3 or 6 recommended doses.Overall,no significant differences were detected in the blood tests or serum biochemistry of the chickens between the treatment groups and the control group.As a result,eugenol essential oil can cure chicken coccidiosis by improving the intestinal microbial structure in the chicken cecum and decreasing the cecum's inflammatory reactions,thus strengthening immune function and eventually demonstrating anticoccidial properties.展开更多
Glycosylphosphatidylinositol(GPI)anchoring represents a fundamental post-translational modification in eukaryotic cells.In fungi,this modification facilitates diverse biological functions through protein targeting to ...Glycosylphosphatidylinositol(GPI)anchoring represents a fundamental post-translational modification in eukaryotic cells.In fungi,this modification facilitates diverse biological functions through protein targeting to the cell wall,yet research on its roles in plant pathogenic fungi remains limited.This study elucidates the function of GPI anchoring in the maize fungal pathogen Cochlibolus heterostrophus.The research demonstrates widespread accumulation of GPI-anchored proteins in hyphae,appressorium and infection hyphae of C.heterostrophus.Deletion of ChGPI7,encoding a crucial enzyme in GPI anchor biosynthesis,substantially reduced vegetative growth,conidiation,and virulence through impaired appressorium formation and invasive growth.The ΔChgpi7 mutants exhibited marked deficiencies in cell wall integrity,leading to decreased stress resistance.Both ChGPI7 deletion and hydro fluoric acid(HF)pyridine treatment eliminated cell wall GPI-anchored proteins and exposed chitin,indicating that GPI-anchored proteins shield chitin from host immune recognition.Analysis identified 124 predicted GPI-anchored proteins in C.heterostrophus,including the putative cell wall glycoprotein ChFEM1.The deletion of ChFEM1 similarly reduced virulence and compromised infection structures and cell wall integrity.Additionally,ChGPI7 influenced both the cell wall localization and protein abundance of ChFEM1.These findings demonstrate that GPI anchoring mediates cell wall integrity and immune evasion during C.heterostrophus infection.展开更多
Sacbrood virus(SBV)is one of the most pathogenic honeybee viruses with host specificity and regional variation.The SBV strain infecting the Chinese honeybee(Apis cerana)is known as Chinese sacbrood virus(CSBV).The ext...Sacbrood virus(SBV)is one of the most pathogenic honeybee viruses with host specificity and regional variation.The SBV strain infecting the Chinese honeybee(Apis cerana)is known as Chinese sacbrood virus(CSBV).The extensively used CSBV detection methods require professionals and expensive equipment;thus,they are unsuitable for rapid onsite CSBV detection.To achieve early and rapid detection of CSBV,we developed a lateral flow detection(LFD)strip method for CSBV detection via clustered regularly interspaced short palindromic repeats(CRISPR)and the Cas13a technique.On the basis of the conserved CSBV VP2 gene nucleotide region,we designed 3 recombinant enzyme-assisted amplification(RAA)primer pairs and prepared 3 corresponding crRNAs.We investigated key performance metrics,including the sensitivity,specificity,and accuracy of LFD strips.The results demonstrated that the LFD strip based on the optimal combination(primer 2+crRNA 2)presented the lowest detection limit(2.80×101 copies/μL),and this strip could complete CSBV detection within 1 h.Furthermore,this strip exhibited excellent detection specificity,with no cross-reactivity with four other honeybee viruses.A test of 100 clinical samples indicated the feasibility of the LFD method for CSBV detection.A comparison of various CSBV detection methods revealed that the CRISPR-Cas13a-based LFD method was more accurate,efficient,and sensitive than the other methods were,indicating great application prospects in onsite CSBV detection.Our developed method is highly important for preventing and controlling CSBV infection as well as maintaining honeybee health.展开更多
The gastrointestinal tract of felines is inhabited by an active and intricate population of microorganisms whose alteration creates disturbances in the immune response and can affect health and disease states.Studies ...The gastrointestinal tract of felines is inhabited by an active and intricate population of microorganisms whose alteration creates disturbances in the immune response and can affect health and disease states.Studies using vari‑ous analytical methods have identified peculiar trends in various illnesses,with Firmicutes being the most prevalent phylum,followed by Bacteroidetes,Proteobacteria,and Actinobacteria.However,more Firmicutes and fewer Bac‑teroidetes have been observed in cats infected with Feline coronavirus.Alterations in the composition of these gut microbiota can be solved by microbiota modification through dietary fiber,probiotics,and fecal microbiota transplan‑tation.Therefore,it is critical to understand the composition of the gut microbiota,the changes in and roles of the gut environment,and the importance of these concepts for overall health while considering the exchange of microbes between humans and domestic animals.This review provides comprehensive information on feline gut microbiota composition,modulation,and analytic methods used for characterizing the gut microbiota.展开更多
Background AFB_(1)-8,9-exo-epoxide(AFBO)is the highly toxic product of Aflatoxin B_(1)(AFB_(1)).Glutathione S-transferases(GSTs)play pivotal roles in detoxifying AFB_(1) by catalyzing the conjugation of AFBO with glut...Background AFB_(1)-8,9-exo-epoxide(AFBO)is the highly toxic product of Aflatoxin B_(1)(AFB_(1)).Glutathione S-transferases(GSTs)play pivotal roles in detoxifying AFB_(1) by catalyzing the conjugation of AFBO with glutathione(GSH).Although there are over 20 GST isozymes that have been identified in chicken,GST isozymes involved in the detoxification process of AFB_(1) have not been identified yet.The objective of this study was to determine which GST isozymes played key role in detoxification of AFB_(1).Results A total of 17 pcDNA3.1(+)-GST isozyme plasmids were constructed and the GST isozyme genes were overexpressed by 80–2,500,000 folds in the chicken Leghorn male hepatoma(LMH)cells.Compared to the AFB_(1) treatment,overexpression of GSTA2X,GSTA3,GSTT1L,GSTZ1-1,and GSTZ1-2 increased the cell viability by 6.5%–17.0%in LMH cells.Moreover,overexpression of five GST isozymes reduced the release of lactate dehydrogenase and reactive oxygen species by 8.8%–64.4%,and 57.2%–77.6%,respectively,as well as enhanced the production AFBO-GSH by 15.8%–19.6%,thus mitigating DNA damage induced by AFB_(1).After comprehensive evaluation of various indicators,GSTA2X displayed the best detoxification effects against AFB_(1).GSTA2X was expressed in Pichia pastoris X-33 and its enzymatic properties for catalyzing the conjugation of AFBO with GSH showed that the optimum temperature and pH were 20–25℃ and 7.6–8.6 as well as the enzymatic kinetic parameter V_(max) was 0.23 nmol/min/mg and the Michaelis constant was 86.05μmol/L with the AFB_(1) as substrate.Conclusions In conclusion,GSTA2X,GSTA3,GSTT1L,GSTZ1-1,and GSTZ1-2 played key roles in AFB_(1) detoxification,which will provide new remediation strategies to prevent aflatoxicosis in chickens.展开更多
Background This study was carried out to investigate the individual and combined contamination of aflatoxin B_(1)(AFB_(1)),deoxynivalenol(DON),and zearalenone(ZEN)in feeds in China between 2021 and 2024.A total of 23,...Background This study was carried out to investigate the individual and combined contamination of aflatoxin B_(1)(AFB_(1)),deoxynivalenol(DON),and zearalenone(ZEN)in feeds in China between 2021 and 2024.A total of 23,003 feed samples,including 17,489 feedstuff samples and 5,514 complete feed samples,were collected from different provinces of China for mycotoxin analysis.Results The analyzed mycotoxins displayed considerably high contamination in the feed samples,with the individual contamination of AFB_(1),DON,and ZEN were 20.0%–100%,33.3%–100%,and 85.0%–100%,respectively.The average concentrations of AFB_(1),DON,and ZEN were 1.2–728.7μg/kg,106–8,634.8μg/kg,and 18.1–3,341.6μg/kg,respectively.Notably,the rates over China’s safety standards for AFB_(1),DON,and ZEN in raw ingredients were 9.7%,2.7%,and 15.7%,respectively.Meanwhile,3.5%,1.1%,and 8.7%of analyzed complete feeds exceeded China’s safety standards for AFB_(1),DON,and ZEN,respectively.Moreover,the co-contamination rates of AFB_(1),DON,and ZEN in more than 70%of raw ingredients and 87.5%of complete feed products were 60.0%–100%and 61.5%–100%,respectively.Conclusion This study reveals that the feeds in China have commonly been contaminated with AFB_(1),DON,and ZEN alone and their combination during the past four years.These findings highlight the significance of monitoring mycotoxin contaminant levels in domestic animal feed and the importance of carrying out feed administration and remediation strategies for mycotoxin control.展开更多
The growth performance of pakchoi （Brassica chinensis L.） in relation to soil cadmium （Cd） fractionations was investigated to evaluate the remediating effect of poultry manure compost on Cd-contaminated soil. A ye...The growth performance of pakchoi （Brassica chinensis L.） in relation to soil cadmium （Cd） fractionations was investigated to evaluate the remediating effect of poultry manure compost on Cd-contaminated soil. A yellow-brown soil （Alfisol） treated with various levels of Cd （0 50 mg Cd kg^-1 soil） was amended with increasing amounts of compost from 0 to 120 g kg^-1. Compost application transformed 47.8%-69.8% of soluble/exchangeable Cd to the organic-bound fraction, and consequently decreased Cd uptake of pakchoi by 56.2%-62.5% as compared with unamended soil. Alleviation of Cd bioavailability by compost was attributed primarily to the increase of soil pH and complexation of Cd by organic matter including dissolved organic matter. In general, the improvement of pakchoi performance was more pronounced in higher Cd-contaminated soil. Addition of large amount of compost also favored the anti-oxidative capability of pakchoi against Cd toxicity. This low cost remediation method seems to be very effective in the restoration of Cd-eontaminated soils.展开更多
β-cryptoxanthin （CRY）, a major carotenoid of potential interest for health, is obtained naturally from orange vegetables and fruits. A few research studies have reported that CRY could decrease oxidative stress and...β-cryptoxanthin （CRY）, a major carotenoid of potential interest for health, is obtained naturally from orange vegetables and fruits. A few research studies have reported that CRY could decrease oxidative stress and germ cell apoptosis. The purpose of this study was to examine the effects of CRY on acute cadmium chloride （CdCl2）-induced oxidative damage in rat testes. For this study, 24 rats were divided into four groups, one of which serves as a control group that received intraperitoneal （i.p.） injections of corn oil and physiological saline. The other rats were i.p. injected with CRY （10 μg kg^-1） every 8 h, beginning 8 h before CdCI2 （2.0 mg kg^-1） treatment. The pathological and TUNEL findings revealed that CRY ameliorated the Cd-induced testicular histological changes and germ cell apoptosis in the rats, Furthermore, the Cd-induced decrease in the testicular testosterone （T） level was attenuated after CRY administration （P 〈 0.05）. The administration of CRY significantly reversed the Cd-induced increases in the lipid peroxide （LPO） and malondialdehyde （MDA） levels （P 〈 0.01）. The testicular antioxidants superoxide dismutase （SOD）, catalase （CAT） and glutathione （GSH） were decreased by treatment with Cd alone but were restored by CRY co-treatment. These results demonstrated that the application of CRY can enhance the tolerance of rats to Cd-induced oxidative damage and suggest that it has promised as a pharmacological agent to protect against Cd-induced testicular toxicity.展开更多
Porcine epidemic diarrhea virus(PEDV),an enteropathogenic coronavirus,has catastrophic impacts on the global pig industry.However,there remain no effective drugs against PEDV infection.In this study,we utilized a reco...Porcine epidemic diarrhea virus(PEDV),an enteropathogenic coronavirus,has catastrophic impacts on the global pig industry.However,there remain no effective drugs against PEDV infection.In this study,we utilized a recombinant PEDV expressing renilla luciferase(PEDV-Rluc)to screen potential anti-PEDV agents from an FDAapproved drug library in Vero cells.Four compounds were identified that significantly decreased luciferase activity of PEDV-Rluc.Among them,niclosamide was further characterized because it exhibited the most potent antiviral activity with the highest selectivity index.It can efficiently inhibit viral RNA synthesis,protein expression and viral progeny production of classical and variant PEDV strains in a dose-dependent manner.Time of addition assay showed that niclosamide exhibited potent anti-PEDV activity when added simultaneously with or after virus infection.Furthermore,niclosamide significantly inhibited the entry stage of PEDV infection by affecting viral internalization rather than viral attachment to cells.In addition,a combination with other small molecule inhibitors of endosomal acidification enhanced the anti-PEDV effect of niclosamide in vitro.Taken together,these findings suggested that niclosamide is a novel antiviral agent that might provide a basis for the development of novel drug therapies against PEDV and other related pathogenic coronavirus infections.展开更多
Porcine epidemic diarrhea virus(PEDV)is the main cause of diarrhea,vomiting,and mortality in pigs,which results in devastating economic loss to the pig industry around the globe.In recent years,the advent of RNAsequen...Porcine epidemic diarrhea virus(PEDV)is the main cause of diarrhea,vomiting,and mortality in pigs,which results in devastating economic loss to the pig industry around the globe.In recent years,the advent of RNAsequencing technologies has led to delineate host responses at late stages of PEDV infection;however,the comparative analysis of host responses to early-stage infection of virulent and avirulent PEDV strains is currently unknown.Here,using the BGI DNBSEQ RNA-sequencing,we performed global gene expression profiles of pig intestinal epithelial cells infected with virulent(GDS01)or avirulent(HX)PEDV strains for 3,6,and 12 h.It was observed that over half of all significantly dysregulated genes in both infection groups exhibited a down-regulated expression pattern.Functional enrichment analyses indicated that the differentially expressed genes(DEGs)in the GDS01 group were predominantly related to autophagy and apoptosis,whereas the genes showing the differential expression in the HX group were strongly enriched in immune responses/inflammation.Among the DEGs,the functional association of TLR3 and IFIT2 genes with the HX and GDS01 strains replication was experimentally validated by TLR3 inhibition and IFIT2 overexpression systems in cultured cells.TLR3 expression was found to inhibit HX strain,but not GDS01 strain,replication by enhancing the IFIT2 expression in infected cells.In conclusion,our study highlights similarities and differences in gene expression patterns and cellular processes/pathways altered at the early-stage infection of PEDV virulent and avirulent strains.These findings may provide a foundation for establishing novel therapies to control PEDV infection.展开更多
Rabies remains a public health threat that kills approximately 59,000 people worldwide each year,most of which are from the developing countries of Africa and Asia where dog rabies are endemic.Therefore, developing an...Rabies remains a public health threat that kills approximately 59,000 people worldwide each year,most of which are from the developing countries of Africa and Asia where dog rabies are endemic.Therefore, developing an affordable and efficacious vaccine is crucial for rabies control in these countries. Interleukin(IL)-15, an immunoregulatory cytokine, is a pluripotent molecule with therapeutic potential, which targets many cell types and links the innate and adaptive immune system. In this study, IL-15 gene was cloned and inserted into the genome of a recombinant rabies virus(RABV) strain LBNSE(designated as LBNSE-IL15), and the effect of over-expression of IL-15 on the immunogenicity of RABV was investigated. It was found that mice vaccinated with LBNSEIL15 could induce significantly higher level of virus-neutralizing antibody(VNA) than those immunized with LBNSE, resulting in the higher protection after challenge. Further investigation was performed to find out the possible role of IL-15 plays in the process of antibody induction, and it was found that LBNSE-IL15 could enhance the maturation of dendritic cells(DCs) in immunized mice. Furthermore, the mice immunized with LBNSE-IL15 could promote the T_(FH) cells differentiation and the generation of germinal center B cells and plasma cells. Together, these data indicated that IL-15 could be a potential adjuvant in enhancing the immunogenicity of RABV, contributing to the development of more-efficacious rabies vaccines.展开更多
The emergence of highly virulent porcine epidemic diarrhea virus(PEDV) variants in China caused huge economic losses in 2010. Since then, large-scale sporadic outbreaks of PED caused by PEDV variants have occasionally...The emergence of highly virulent porcine epidemic diarrhea virus(PEDV) variants in China caused huge economic losses in 2010. Since then, large-scale sporadic outbreaks of PED caused by PEDV variants have occasionally occurred in China. However, the molecular diversity and epidemiology of PEDV in different provinces has not been completely understood. To determine the molecular diversity of PEDV in the Hubei Province of China, we collected 172 PED samples from 34 farms across the province in 2016 and performed reverse transcription polymerase chain reaction(RTPCR)by targeting the nucleocapsid(N) gene. Seventy-four samples were found to be PEDVpositive.We further characterized the complete spike(S) glycoprotein genes from the positive samples and found 21 different S genes with amino acid mutations. The PEDV isolates here presented most of the genotypes which were found previously in field isolates in East and SouthEast Asia, North America, and Europe. Besides the typical Genotypes Ⅰ and Ⅱ, the INDEX groups were also found. Importantly, 58 new amino acids mutant sites in the S genes, including 44 sites in S1 and 14 sites in S2, were first described. Our results revealed that the S genes of PEDV showed variation and that diverse genotypes of PEDV coexisted and were responsible for the PED outbreaks in Hubei in 2016. This work highlighted the complexity of the epidemiology of PEDV and emphasized the need for reassessing the efficacy of classic PEDV vaccines against emerging variant strains and developing new vaccines to facilitate the prevention and control of PEDV in fields.展开更多
African swine fever(ASF)is an infectious disease caused by African swine fever virus(ASFV)with clinical symptoms of high fever,hemorrhages and high mortality rate,posing a threat to the global swine industry and food ...African swine fever(ASF)is an infectious disease caused by African swine fever virus(ASFV)with clinical symptoms of high fever,hemorrhages and high mortality rate,posing a threat to the global swine industry and food security.Quarantine and control of ASFV is crucial for preventing swine industry from ASFV infection.In this study,a recombinase polymerase amplification(RPA)-CRISPR-based nucleic acid detection method was developed for diagnosing ASF.As a highly sensitive method,RPA-CRISPR can detect even a single copy of ASFV plasmid and genomic DNA by determining fluorescence signal induced by collateral cleavage of CRISPR-lw Cas13 a(previously known as C2c2)through quantitative real-time PCR(q PCR)and has the same or even higher sensitivity than the traditional q PCR method.A lateral flow strip was developed and used in combination with RPA-CRISPR for ASFV detection with the same level of sensitivity of Taq Man q PCR.Likewise,RPA-CRISPR is capable of distinguishing ASFV genomic DNA from viral DNA/RNA of other porcine viruses without any cross-reactivity.This diagnostic method is also available for diagnosing ASFV clinical DNA samples with coincidence rate of 100%for both ASFV positive and negative samples.RPA-CRISPR has great potential for clinical quarantine of ASFV in swine industry and food security.展开更多
Rabies virus(RABV)is an infectious and neurotropic pathogen that causes rabies and infects humans and almost all warm-blooded animals,posing a great threat to people and public safety.It is well known that innate immu...Rabies virus(RABV)is an infectious and neurotropic pathogen that causes rabies and infects humans and almost all warm-blooded animals,posing a great threat to people and public safety.It is well known that innate immunity is the critical first line of host defense against viral infection.It monitors the invading pathogens by recognizing the pathogen-associated molecular patterns and danger-associated molecular patterns through pattern-recognition receptors,leading to the production of type I interferons(IFNα/β),inflammatory cytokines,and chemokines,or the activation of autophagy or apoptosis to inhibit virus replication.In the case of RABV,the innate immune response is usually triggered when the skin or muscle is bitten or scratched.However,RABV has evolved many ways to escape or even hijack innate immune response to complete its own replication and eventually invades the central nervous system(CNS).Once RABV reaches the CNS,it cannot be wiped out by the immune system or any drugs.Therefore,a better understanding of the interplay between RABV and innate immu-nity is necessary to develop effective strategies to combat its infection.Here,we review the innate immune responses induced by RABV and illustrate the antagonism mechanisms of RABV to provide new insights for the control of rabies.展开更多
Background:The hyper-prolificacy Meishan gilts achieved a superior endometrial gland development(EGD)than white crossbred gilts during the ovary-independent period(before 60 d of age).Then,the EGD continues under the ...Background:The hyper-prolificacy Meishan gilts achieved a superior endometrial gland development(EGD)than white crossbred gilts during the ovary-independent period(before 60 d of age).Then,the EGD continues under the management of ovary-derived steroid hormones that regulated by gut microbiota(after 60 d of age).However,whether Meishan gilts’superiority in EGD lasting to the ovary-dependent period(after 60 d of age)and the role of gut microbiota in this period both remain unclear.Methods:Meishan gilts and Landrace x Yorkshire(LxY)gilts were raised under the same housing and feeding conditions until sexual maturity and then we compared their EGD and gut microbiota.Meanwhile,we transplanted fecal microbiota from Meishan gilts to L×Y gilts to explore the role of gut microbiota in EGD.We sampled plasma every 3 weeks and collected the uterus,ovary,liver,and rectal feces after the sacrifice.We then determined the hormone concentrations and expressions of the EGD-related genes.We also profiled the gut microbiota using 16S rDNA sequencing and metabolites of plasma and liver tissue using untargeted metabolomics.Finally,the correlation analysis and significant test was conducted between FMT-shifted gut microbes and EGD-related indices.Results:Meishan gilts have larger endometrial gland area(P<0.001),longer uterine horn length(P<0.01)but lighter uterine horn weight(P<0.05),a distinctive gut microbiota compared with L×Y gilts.Fecal microbiota transplantation(FMT)increased endometrial gland area(P<0.01).FMT markedly shifted the metabolite profiles of both liver and plasma,and these differential metabolites enriched in steroid hormone biosynthesis pathway.FMT increased estradiol and insulin-like growth factor 1 but decreased progesterone dynamically.FMT also increased the expression of the EGD-related genes estrogen receptor 1 gene,epithelial cadherin,and forkhead box protein A2.There is a significant correlation between FMT-shifted gut microbes and EGD-related indices.Conclusion:Sexually matured Meishan gilts achieved a superior EGD than LxY gilts.Meanwhile,gut microbiota contribute to the EGD potentially via regulating of steroid hormones during the ovary-dependent period.展开更多
The adsorption and desorption of the toxin from Bacillus thuringiensis strain WG-001 on rectorite were studied at different toxin and/or rectorite concentrations, pH values and temperatures. The insecticidal activity ...The adsorption and desorption of the toxin from Bacillus thuringiensis strain WG-001 on rectorite were studied at different toxin and/or rectorite concentrations, pH values and temperatures. The insecticidal activity of the adsorbed toxin was evaluated by determining the lethal concentration to kill 50% of the larvae of Heliothis armigera （LC50）. The adsorption of the toxin on rectorite in sodium carbonate buffer （pH 9） reached equilibrium within 0.5-1.0 h and the adsorption isotherm of the toxin followed the Langmuir equation （R^2 〉 0.99）. In the pH range from 9 to 11 （carbonate buffer）, the adsorbed toxin decreased with increasing pH. The adsorption amounts decreased with increasing rectorite：toxin ratio. The adsorption was not significantly affected by the temperature between 10 and 50 ℃. The X- ray diffraction analysis indicated occurrence of the intercalation of the rectorite by the toxin. The infrared absorption spectrum showed that the binding of the toxin did not alter its structure. The LC50 wlues of the adsorbed toxin were smaller than those of the free toxin. The rectorite protected the toxin from ultraviolet irradiation damage. The desorption of the adsorbed toxin in water ranged from 37.5% to 56.4% and from 27.4% to 41.8% in a carbonate buffer. The desorption percentage also decreased with increasing rectorite：toxin ratio.展开更多
Suppressive regulatory T cells(Treg cells)play a vital role in preventing autoimmunity and restraining excessive immune response to both self-and non-self-antigens.Studies on humans and mice show that the Forkhead box...Suppressive regulatory T cells(Treg cells)play a vital role in preventing autoimmunity and restraining excessive immune response to both self-and non-self-antigens.Studies on humans and mice show that the Forkhead box p3(Foxp3)is a key regulatory gene for the development and function of Treg cells.In zebrafish,Treg cells have been identified by using foxp3a as a reliable marker.However,little is known about the function of foxp3a and Treg cells in gonadal development and sex differentiation.Here,we show that foxp3a is essential for maintaining immune homeostasis in zebrafish testis development.We found that foxp3a was specifically expressed in a subset of T cells in zebrafish testis,while knockout of foxp3a led to deficiency of foxp3a-positive Treg cells in the testis.More than 80%of foxp3a^(-/-)mutants developed as subfertile males,and the rest of the mutants developed as fertile females with decreased ovulation.Further study revealed that foxp3a^(-/-)mutants had a delayed juvenile ovary-to-testis transition in definite males and sex reversal in about half of the definite females,which led to a dominance of later male development.Owing to the absence of foxp3a-positive Treg cells in the differentiating testis of foxp3a^(-/-)mutants,abundant T cells and macrophages expand to disrupt an immunosuppressive milieu,resulting in defective development of germ cells and gonadal somatic cells and leading to development of infertile males.Therefore,our study reveals that foxp3a-positive Treg cells play an essential role in the orchestration of gonadal development and sex differentiation in zebrafish.展开更多
Root-knot nematodes(RKNs) cause huge yield losses to agricultural crops worldwide. Meanwhile, livestock manure is often improperly managed by farmers, which leads to serious environmental pollution. To resolve these t...Root-knot nematodes(RKNs) cause huge yield losses to agricultural crops worldwide. Meanwhile, livestock manure is often improperly managed by farmers, which leads to serious environmental pollution. To resolve these two problems, this study developed a procedure for the conversion of chicken manure to organic fertilizer by larvae of Hermetia illucens L. and Bacillus subtilis BSF-CL. Chicken manure organic fertilizer was then mixed thoroughly with Paenibacillus polymyxa KM2501-1 to a final concentration of 1.5×10^(8)CFU g^(-1). The efficacy of KM2501-1 microbial organic fertilizer in controlling root-knot nematodes was evaluated in pot and field experiments. In pot experiments, applying KM2501-1 microbial organic fertilizer either as a base fertilizer or as a fumigant at the dose of 40 g/pot suppressed root-knot disease by 61.76 and 69.05% compared to the corresponding control treatments, respectively. When applied as a fumigant at the dose of 1 kg m;in field experiments, KM2501-1 microbial organic fertilizer enhanced the growth of tomato plants, suppressed root-knot disease by 49.97%, and reduced second stage juveniles of RKN in soil by 88.68%. KM2501-1 microbial organic fertilizer controlled RKNs better than commercial bio-organic fertilizer in both pot and field experiments. These results demonstrate that this co-conversion process efficiently transforms chicken manure into high value-added larvae biomass and KM2501-1 microbial organic fertilizer with potential application as a novel nematode control agent.展开更多
基金supported by the Fundamental Research Funds for the Central Universities in China(Grant no.2662022DKPY003)the HZAU-AGIS Cooperation Fund(Grant no.SZYJY2022015).
文摘Toxoplasma gondii(T.gondii)is an obligate intracellular parasite with a wide range of hosts,including humans and many warm-blooded animals.The parasite exists in two interconvertible forms,namely tachyzoites and bradyzoites in intermediate hosts that are responsible for acute and chronic infections respectively.Mature bradyzoites accumulate large amounts of amylopectin granules but their roles have not been fully characterized.In this study,the predicted key enzymes involved in amylopectin synthesis(UDP-sugar pyrophospharylase,USP)and degradation(alpha-glucan water dikinase,GWD)of ME49 strain were individually knocked out,and then bradyzoite-related phenotyping experiments in vitro and in vivo were performed to dissect their roles during parasite growth and development.Deletion of the usp or gwd gene in the type II strain ME49 reduced the replication rates of tachyzoites in vitro and parasite virulence in vivo,suggesting that amylopectin metabolism is important for optimal tachyzoite growth.Interestingly,the Δusp mutant grew slightly faster than the parental strain under stress conditions that induced bradyzoite transition,which was likely due to the decreased efficiency of bradyzoite formation of theΔusp mutant.Although theΔgwd mutant could convert to bradyzoite robustly in vitro,it was significantly impaired in establishing chronic infection in vivo.Both the Δusp and Δgwd mutants showed a dramatic reduction in the reactivation of chronic infection in an in vitro model.Together,these results suggest that USP and GWD,which are involved in amylopectin synthesis and degradation have important roles in tachyzoite growth,as well as in the formation and reactivation of bradyzoites in T.gondii.
基金supported by National Natural Science Foundation of China (Nos. 82173774, 31925037, 22104041)the Fundamental Research Funds for the Central Universities (Nos 2662021DKQD001, 2662021JC001)the Cheeloo Scholar Program of Shandong University (to W. Liu)。
文摘Advances in microbiology rely on innovations in technology. Droplet microfluidics, as a versatile and powerful technique that allows high-throughput generation and manipulation of subnanoliter volume droplets, has become an indispensable tool shifting experimental paradigms in microbiology. Droplet microfluidics has opened new avenues to various microbiological research, from resolving single-cell heterogeneity to investigating spatiotemporal dynamics of microbial communities, from precise quantitation of microbiota to systematic decipherment of microbial interactions, and from isolating rare and uncultured microbes to improving genetic engineered strains. In this review, we present recent advances of droplet microfluidics in various fields of microbiology: i) microbial cultivation, ii) microorganism detection and characterization, iii) antibiotic susceptibility testing, iv) microbial interactions, v) microbial biotechnology.We also provide our perspectives on the challenges and future directions for droplet microfluidic-based microbiology research.
基金supported by the National Key Research and Development Program(2016YFD0501303).
文摘The development of alternative therapies to treat chicken coccidiosis has become a hot topic because of the widespread use of conventional medicines.This study aimed to investigate the effectiveness of eugenol in treating Eimeria tenella infection in broilers.Broiers,at the age of 14 d,were orally infected with sporulated Eimeria tenella oocysts,and then,eugenol essential oil was added to chicken feed at three different dosages(0.1,0.2 or 0.4 g/kg).The anticoccidial effects of eugenol essential oil were assessed using the anticoccidial index(ACI).As a result,eugenol exhibited a moderate anticoccidial effect,with an ACI of 167.37 at 0.2 g/kg.After eugenol treatment,the expression of occludin in the epithelial cells of the chicken cecum was significantly greater(P<0.05)than that in the epithelial cells of the nontreated control(IC)group.The proportion of intestinal Lactobacillus_agilli increased.Eugenol therapy dramatically increased the activity of superoxide dismutase.After high-dose treatment,the expression of the proinflammatory factors IL-1βand IL-6 significantly decreased,while the expression of the cytokines IL-4 and IFN-γsignificantly increased.The safety of eugenol essential oil was evaluated at the 1,3 or 6 recommended doses.Overall,no significant differences were detected in the blood tests or serum biochemistry of the chickens between the treatment groups and the control group.As a result,eugenol essential oil can cure chicken coccidiosis by improving the intestinal microbial structure in the chicken cecum and decreasing the cecum's inflammatory reactions,thus strengthening immune function and eventually demonstrating anticoccidial properties.
基金supported by the Fundamental Research Funds for the Central Universities,China(2021ZKPY007).
文摘Glycosylphosphatidylinositol(GPI)anchoring represents a fundamental post-translational modification in eukaryotic cells.In fungi,this modification facilitates diverse biological functions through protein targeting to the cell wall,yet research on its roles in plant pathogenic fungi remains limited.This study elucidates the function of GPI anchoring in the maize fungal pathogen Cochlibolus heterostrophus.The research demonstrates widespread accumulation of GPI-anchored proteins in hyphae,appressorium and infection hyphae of C.heterostrophus.Deletion of ChGPI7,encoding a crucial enzyme in GPI anchor biosynthesis,substantially reduced vegetative growth,conidiation,and virulence through impaired appressorium formation and invasive growth.The ΔChgpi7 mutants exhibited marked deficiencies in cell wall integrity,leading to decreased stress resistance.Both ChGPI7 deletion and hydro fluoric acid(HF)pyridine treatment eliminated cell wall GPI-anchored proteins and exposed chitin,indicating that GPI-anchored proteins shield chitin from host immune recognition.Analysis identified 124 predicted GPI-anchored proteins in C.heterostrophus,including the putative cell wall glycoprotein ChFEM1.The deletion of ChFEM1 similarly reduced virulence and compromised infection structures and cell wall integrity.Additionally,ChGPI7 influenced both the cell wall localization and protein abundance of ChFEM1.These findings demonstrate that GPI anchoring mediates cell wall integrity and immune evasion during C.heterostrophus infection.
基金supported by the National Key Research and Development Program of China(NO.2022YFD1600202)the Lush Mountain Public Welfare Protection Action of the China Environmental Protection Foundation(CEPFQS202169-14).
文摘Sacbrood virus(SBV)is one of the most pathogenic honeybee viruses with host specificity and regional variation.The SBV strain infecting the Chinese honeybee(Apis cerana)is known as Chinese sacbrood virus(CSBV).The extensively used CSBV detection methods require professionals and expensive equipment;thus,they are unsuitable for rapid onsite CSBV detection.To achieve early and rapid detection of CSBV,we developed a lateral flow detection(LFD)strip method for CSBV detection via clustered regularly interspaced short palindromic repeats(CRISPR)and the Cas13a technique.On the basis of the conserved CSBV VP2 gene nucleotide region,we designed 3 recombinant enzyme-assisted amplification(RAA)primer pairs and prepared 3 corresponding crRNAs.We investigated key performance metrics,including the sensitivity,specificity,and accuracy of LFD strips.The results demonstrated that the LFD strip based on the optimal combination(primer 2+crRNA 2)presented the lowest detection limit(2.80×101 copies/μL),and this strip could complete CSBV detection within 1 h.Furthermore,this strip exhibited excellent detection specificity,with no cross-reactivity with four other honeybee viruses.A test of 100 clinical samples indicated the feasibility of the LFD method for CSBV detection.A comparison of various CSBV detection methods revealed that the CRISPR-Cas13a-based LFD method was more accurate,efficient,and sensitive than the other methods were,indicating great application prospects in onsite CSBV detection.Our developed method is highly important for preventing and controlling CSBV infection as well as maintaining honeybee health.
基金funded this manuscript for the Central Universities(Grant Nos.2662023DKPY004 and 2662021DKQD005).
文摘The gastrointestinal tract of felines is inhabited by an active and intricate population of microorganisms whose alteration creates disturbances in the immune response and can affect health and disease states.Studies using vari‑ous analytical methods have identified peculiar trends in various illnesses,with Firmicutes being the most prevalent phylum,followed by Bacteroidetes,Proteobacteria,and Actinobacteria.However,more Firmicutes and fewer Bac‑teroidetes have been observed in cats infected with Feline coronavirus.Alterations in the composition of these gut microbiota can be solved by microbiota modification through dietary fiber,probiotics,and fecal microbiota transplan‑tation.Therefore,it is critical to understand the composition of the gut microbiota,the changes in and roles of the gut environment,and the importance of these concepts for overall health while considering the exchange of microbes between humans and domestic animals.This review provides comprehensive information on feline gut microbiota composition,modulation,and analytic methods used for characterizing the gut microbiota.
基金supported by the Chinese Natural Science Foundation Projects 32072775,32272915 and 32472949the National Key Research and Development Programs of China(2023YFD1301003 and 2023YFD1301005)the Fundamental Research Funds for the Central Universities(2662023DKPY002)。
文摘Background AFB_(1)-8,9-exo-epoxide(AFBO)is the highly toxic product of Aflatoxin B_(1)(AFB_(1)).Glutathione S-transferases(GSTs)play pivotal roles in detoxifying AFB_(1) by catalyzing the conjugation of AFBO with glutathione(GSH).Although there are over 20 GST isozymes that have been identified in chicken,GST isozymes involved in the detoxification process of AFB_(1) have not been identified yet.The objective of this study was to determine which GST isozymes played key role in detoxification of AFB_(1).Results A total of 17 pcDNA3.1(+)-GST isozyme plasmids were constructed and the GST isozyme genes were overexpressed by 80–2,500,000 folds in the chicken Leghorn male hepatoma(LMH)cells.Compared to the AFB_(1) treatment,overexpression of GSTA2X,GSTA3,GSTT1L,GSTZ1-1,and GSTZ1-2 increased the cell viability by 6.5%–17.0%in LMH cells.Moreover,overexpression of five GST isozymes reduced the release of lactate dehydrogenase and reactive oxygen species by 8.8%–64.4%,and 57.2%–77.6%,respectively,as well as enhanced the production AFBO-GSH by 15.8%–19.6%,thus mitigating DNA damage induced by AFB_(1).After comprehensive evaluation of various indicators,GSTA2X displayed the best detoxification effects against AFB_(1).GSTA2X was expressed in Pichia pastoris X-33 and its enzymatic properties for catalyzing the conjugation of AFBO with GSH showed that the optimum temperature and pH were 20–25℃ and 7.6–8.6 as well as the enzymatic kinetic parameter V_(max) was 0.23 nmol/min/mg and the Michaelis constant was 86.05μmol/L with the AFB_(1) as substrate.Conclusions In conclusion,GSTA2X,GSTA3,GSTT1L,GSTZ1-1,and GSTZ1-2 played key roles in AFB_(1) detoxification,which will provide new remediation strategies to prevent aflatoxicosis in chickens.
基金supported by the Chinese Natural Science Foundation projects(32272915 and 32472949)National Key Research and Development Programs of China(2023YFD1301003)+1 种基金the Fundamental Research Funds for the Central Universities(2662023DKPY002)Hebei Panshuo Biotechnolog Co.,Ltd.
文摘Background This study was carried out to investigate the individual and combined contamination of aflatoxin B_(1)(AFB_(1)),deoxynivalenol(DON),and zearalenone(ZEN)in feeds in China between 2021 and 2024.A total of 23,003 feed samples,including 17,489 feedstuff samples and 5,514 complete feed samples,were collected from different provinces of China for mycotoxin analysis.Results The analyzed mycotoxins displayed considerably high contamination in the feed samples,with the individual contamination of AFB_(1),DON,and ZEN were 20.0%–100%,33.3%–100%,and 85.0%–100%,respectively.The average concentrations of AFB_(1),DON,and ZEN were 1.2–728.7μg/kg,106–8,634.8μg/kg,and 18.1–3,341.6μg/kg,respectively.Notably,the rates over China’s safety standards for AFB_(1),DON,and ZEN in raw ingredients were 9.7%,2.7%,and 15.7%,respectively.Meanwhile,3.5%,1.1%,and 8.7%of analyzed complete feeds exceeded China’s safety standards for AFB_(1),DON,and ZEN,respectively.Moreover,the co-contamination rates of AFB_(1),DON,and ZEN in more than 70%of raw ingredients and 87.5%of complete feed products were 60.0%–100%and 61.5%–100%,respectively.Conclusion This study reveals that the feeds in China have commonly been contaminated with AFB_(1),DON,and ZEN alone and their combination during the past four years.These findings highlight the significance of monitoring mycotoxin contaminant levels in domestic animal feed and the importance of carrying out feed administration and remediation strategies for mycotoxin control.
基金Project supported by the National High Technology Research and Development Program (863 Program) of China(No. 2006AA06Z350)
文摘The growth performance of pakchoi （Brassica chinensis L.） in relation to soil cadmium （Cd） fractionations was investigated to evaluate the remediating effect of poultry manure compost on Cd-contaminated soil. A yellow-brown soil （Alfisol） treated with various levels of Cd （0 50 mg Cd kg^-1 soil） was amended with increasing amounts of compost from 0 to 120 g kg^-1. Compost application transformed 47.8%-69.8% of soluble/exchangeable Cd to the organic-bound fraction, and consequently decreased Cd uptake of pakchoi by 56.2%-62.5% as compared with unamended soil. Alleviation of Cd bioavailability by compost was attributed primarily to the increase of soil pH and complexation of Cd by organic matter including dissolved organic matter. In general, the improvement of pakchoi performance was more pronounced in higher Cd-contaminated soil. Addition of large amount of compost also favored the anti-oxidative capability of pakchoi against Cd toxicity. This low cost remediation method seems to be very effective in the restoration of Cd-eontaminated soils.
文摘β-cryptoxanthin （CRY）, a major carotenoid of potential interest for health, is obtained naturally from orange vegetables and fruits. A few research studies have reported that CRY could decrease oxidative stress and germ cell apoptosis. The purpose of this study was to examine the effects of CRY on acute cadmium chloride （CdCl2）-induced oxidative damage in rat testes. For this study, 24 rats were divided into four groups, one of which serves as a control group that received intraperitoneal （i.p.） injections of corn oil and physiological saline. The other rats were i.p. injected with CRY （10 μg kg^-1） every 8 h, beginning 8 h before CdCI2 （2.0 mg kg^-1） treatment. The pathological and TUNEL findings revealed that CRY ameliorated the Cd-induced testicular histological changes and germ cell apoptosis in the rats, Furthermore, the Cd-induced decrease in the testicular testosterone （T） level was attenuated after CRY administration （P 〈 0.05）. The administration of CRY significantly reversed the Cd-induced increases in the lipid peroxide （LPO） and malondialdehyde （MDA） levels （P 〈 0.01）. The testicular antioxidants superoxide dismutase （SOD）, catalase （CAT） and glutathione （GSH） were decreased by treatment with Cd alone but were restored by CRY co-treatment. These results demonstrated that the application of CRY can enhance the tolerance of rats to Cd-induced oxidative damage and suggest that it has promised as a pharmacological agent to protect against Cd-induced testicular toxicity.
基金supported by the National Natural Science Foundation of China(Grant No.31602033 and 32172839)the China Scholarship Council under grant 201908410129.
文摘Porcine epidemic diarrhea virus(PEDV),an enteropathogenic coronavirus,has catastrophic impacts on the global pig industry.However,there remain no effective drugs against PEDV infection.In this study,we utilized a recombinant PEDV expressing renilla luciferase(PEDV-Rluc)to screen potential anti-PEDV agents from an FDAapproved drug library in Vero cells.Four compounds were identified that significantly decreased luciferase activity of PEDV-Rluc.Among them,niclosamide was further characterized because it exhibited the most potent antiviral activity with the highest selectivity index.It can efficiently inhibit viral RNA synthesis,protein expression and viral progeny production of classical and variant PEDV strains in a dose-dependent manner.Time of addition assay showed that niclosamide exhibited potent anti-PEDV activity when added simultaneously with or after virus infection.Furthermore,niclosamide significantly inhibited the entry stage of PEDV infection by affecting viral internalization rather than viral attachment to cells.In addition,a combination with other small molecule inhibitors of endosomal acidification enhanced the anti-PEDV effect of niclosamide in vitro.Taken together,these findings suggested that niclosamide is a novel antiviral agent that might provide a basis for the development of novel drug therapies against PEDV and other related pathogenic coronavirus infections.
基金funded by National Key Research and Development Program of China(No.2016YFD0500101)China Postdoctoral Science Foundation(No.2020M672974)
文摘Porcine epidemic diarrhea virus(PEDV)is the main cause of diarrhea,vomiting,and mortality in pigs,which results in devastating economic loss to the pig industry around the globe.In recent years,the advent of RNAsequencing technologies has led to delineate host responses at late stages of PEDV infection;however,the comparative analysis of host responses to early-stage infection of virulent and avirulent PEDV strains is currently unknown.Here,using the BGI DNBSEQ RNA-sequencing,we performed global gene expression profiles of pig intestinal epithelial cells infected with virulent(GDS01)or avirulent(HX)PEDV strains for 3,6,and 12 h.It was observed that over half of all significantly dysregulated genes in both infection groups exhibited a down-regulated expression pattern.Functional enrichment analyses indicated that the differentially expressed genes(DEGs)in the GDS01 group were predominantly related to autophagy and apoptosis,whereas the genes showing the differential expression in the HX group were strongly enriched in immune responses/inflammation.Among the DEGs,the functional association of TLR3 and IFIT2 genes with the HX and GDS01 strains replication was experimentally validated by TLR3 inhibition and IFIT2 overexpression systems in cultured cells.TLR3 expression was found to inhibit HX strain,but not GDS01 strain,replication by enhancing the IFIT2 expression in infected cells.In conclusion,our study highlights similarities and differences in gene expression patterns and cellular processes/pathways altered at the early-stage infection of PEDV virulent and avirulent strains.These findings may provide a foundation for establishing novel therapies to control PEDV infection.
基金partially supported by the National Natural Science Foundation of China(31402176,31372419,31522057)the National Program on Key Research Project of China(2016YFD0500400)+2 种基金the Fundamental Research Funds for the Central Universities(2662016 QD036,to MZ)the Ministry of Science and Technology of China(863 program,number 2011AA10A212)the Ministry of Agriculture of China(special fund for Agro-scientific research in the Public Interest,2013 03042,to ZFF)
文摘Rabies remains a public health threat that kills approximately 59,000 people worldwide each year,most of which are from the developing countries of Africa and Asia where dog rabies are endemic.Therefore, developing an affordable and efficacious vaccine is crucial for rabies control in these countries. Interleukin(IL)-15, an immunoregulatory cytokine, is a pluripotent molecule with therapeutic potential, which targets many cell types and links the innate and adaptive immune system. In this study, IL-15 gene was cloned and inserted into the genome of a recombinant rabies virus(RABV) strain LBNSE(designated as LBNSE-IL15), and the effect of over-expression of IL-15 on the immunogenicity of RABV was investigated. It was found that mice vaccinated with LBNSEIL15 could induce significantly higher level of virus-neutralizing antibody(VNA) than those immunized with LBNSE, resulting in the higher protection after challenge. Further investigation was performed to find out the possible role of IL-15 plays in the process of antibody induction, and it was found that LBNSE-IL15 could enhance the maturation of dendritic cells(DCs) in immunized mice. Furthermore, the mice immunized with LBNSE-IL15 could promote the T_(FH) cells differentiation and the generation of germinal center B cells and plasma cells. Together, these data indicated that IL-15 could be a potential adjuvant in enhancing the immunogenicity of RABV, contributing to the development of more-efficacious rabies vaccines.
基金funded by the National Natural Science Foundation of China(31470260 and 81401672)the"Fundamental Research Funds for the Central Universities"(531107040975)
文摘The emergence of highly virulent porcine epidemic diarrhea virus(PEDV) variants in China caused huge economic losses in 2010. Since then, large-scale sporadic outbreaks of PED caused by PEDV variants have occasionally occurred in China. However, the molecular diversity and epidemiology of PEDV in different provinces has not been completely understood. To determine the molecular diversity of PEDV in the Hubei Province of China, we collected 172 PED samples from 34 farms across the province in 2016 and performed reverse transcription polymerase chain reaction(RTPCR)by targeting the nucleocapsid(N) gene. Seventy-four samples were found to be PEDVpositive.We further characterized the complete spike(S) glycoprotein genes from the positive samples and found 21 different S genes with amino acid mutations. The PEDV isolates here presented most of the genotypes which were found previously in field isolates in East and SouthEast Asia, North America, and Europe. Besides the typical Genotypes Ⅰ and Ⅱ, the INDEX groups were also found. Importantly, 58 new amino acids mutant sites in the S genes, including 44 sites in S1 and 14 sites in S2, were first described. Our results revealed that the S genes of PEDV showed variation and that diverse genotypes of PEDV coexisted and were responsible for the PED outbreaks in Hubei in 2016. This work highlighted the complexity of the epidemiology of PEDV and emphasized the need for reassessing the efficacy of classic PEDV vaccines against emerging variant strains and developing new vaccines to facilitate the prevention and control of PEDV in fields.
基金supported by the National Natural Science Foundation of China(31522057 and 31872451 to LZ,31720103917 and 31872452 to ZF)。
文摘African swine fever(ASF)is an infectious disease caused by African swine fever virus(ASFV)with clinical symptoms of high fever,hemorrhages and high mortality rate,posing a threat to the global swine industry and food security.Quarantine and control of ASFV is crucial for preventing swine industry from ASFV infection.In this study,a recombinase polymerase amplification(RPA)-CRISPR-based nucleic acid detection method was developed for diagnosing ASF.As a highly sensitive method,RPA-CRISPR can detect even a single copy of ASFV plasmid and genomic DNA by determining fluorescence signal induced by collateral cleavage of CRISPR-lw Cas13 a(previously known as C2c2)through quantitative real-time PCR(q PCR)and has the same or even higher sensitivity than the traditional q PCR method.A lateral flow strip was developed and used in combination with RPA-CRISPR for ASFV detection with the same level of sensitivity of Taq Man q PCR.Likewise,RPA-CRISPR is capable of distinguishing ASFV genomic DNA from viral DNA/RNA of other porcine viruses without any cross-reactivity.This diagnostic method is also available for diagnosing ASFV clinical DNA samples with coincidence rate of 100%for both ASFV positive and negative samples.RPA-CRISPR has great potential for clinical quarantine of ASFV in swine industry and food security.
基金National Natural Science Foundation of China,Grant/Award Number:31872487。
文摘Rabies virus(RABV)is an infectious and neurotropic pathogen that causes rabies and infects humans and almost all warm-blooded animals,posing a great threat to people and public safety.It is well known that innate immunity is the critical first line of host defense against viral infection.It monitors the invading pathogens by recognizing the pathogen-associated molecular patterns and danger-associated molecular patterns through pattern-recognition receptors,leading to the production of type I interferons(IFNα/β),inflammatory cytokines,and chemokines,or the activation of autophagy or apoptosis to inhibit virus replication.In the case of RABV,the innate immune response is usually triggered when the skin or muscle is bitten or scratched.However,RABV has evolved many ways to escape or even hijack innate immune response to complete its own replication and eventually invades the central nervous system(CNS).Once RABV reaches the CNS,it cannot be wiped out by the immune system or any drugs.Therefore,a better understanding of the interplay between RABV and innate immu-nity is necessary to develop effective strategies to combat its infection.Here,we review the innate immune responses induced by RABV and illustrate the antagonism mechanisms of RABV to provide new insights for the control of rabies.
基金supported by the Natural Science Foundation of China(31730090 and 31925037)Hubei Provincial Natural Science Foundation of China(2018CFA020).
文摘Background:The hyper-prolificacy Meishan gilts achieved a superior endometrial gland development(EGD)than white crossbred gilts during the ovary-independent period(before 60 d of age).Then,the EGD continues under the management of ovary-derived steroid hormones that regulated by gut microbiota(after 60 d of age).However,whether Meishan gilts’superiority in EGD lasting to the ovary-dependent period(after 60 d of age)and the role of gut microbiota in this period both remain unclear.Methods:Meishan gilts and Landrace x Yorkshire(LxY)gilts were raised under the same housing and feeding conditions until sexual maturity and then we compared their EGD and gut microbiota.Meanwhile,we transplanted fecal microbiota from Meishan gilts to L×Y gilts to explore the role of gut microbiota in EGD.We sampled plasma every 3 weeks and collected the uterus,ovary,liver,and rectal feces after the sacrifice.We then determined the hormone concentrations and expressions of the EGD-related genes.We also profiled the gut microbiota using 16S rDNA sequencing and metabolites of plasma and liver tissue using untargeted metabolomics.Finally,the correlation analysis and significant test was conducted between FMT-shifted gut microbes and EGD-related indices.Results:Meishan gilts have larger endometrial gland area(P<0.001),longer uterine horn length(P<0.01)but lighter uterine horn weight(P<0.05),a distinctive gut microbiota compared with L×Y gilts.Fecal microbiota transplantation(FMT)increased endometrial gland area(P<0.01).FMT markedly shifted the metabolite profiles of both liver and plasma,and these differential metabolites enriched in steroid hormone biosynthesis pathway.FMT increased estradiol and insulin-like growth factor 1 but decreased progesterone dynamically.FMT also increased the expression of the EGD-related genes estrogen receptor 1 gene,epithelial cadherin,and forkhead box protein A2.There is a significant correlation between FMT-shifted gut microbes and EGD-related indices.Conclusion:Sexually matured Meishan gilts achieved a superior EGD than LxY gilts.Meanwhile,gut microbiota contribute to the EGD potentially via regulating of steroid hormones during the ovary-dependent period.
基金the National Natural Science Foundation of China (Nos.40271064 and 30170032)the Natural Science Foundation of Tianjin,China (No.05YFJMJC05100)+1 种基金the Scientific Research Project of the Education Administration of Hubei Province,China (No.Q200611007)the Key Research Program of Wuhan University of Science and Technology,China (No.2006XZ4).
文摘The adsorption and desorption of the toxin from Bacillus thuringiensis strain WG-001 on rectorite were studied at different toxin and/or rectorite concentrations, pH values and temperatures. The insecticidal activity of the adsorbed toxin was evaluated by determining the lethal concentration to kill 50% of the larvae of Heliothis armigera （LC50）. The adsorption of the toxin on rectorite in sodium carbonate buffer （pH 9） reached equilibrium within 0.5-1.0 h and the adsorption isotherm of the toxin followed the Langmuir equation （R^2 〉 0.99）. In the pH range from 9 to 11 （carbonate buffer）, the adsorbed toxin decreased with increasing pH. The adsorption amounts decreased with increasing rectorite：toxin ratio. The adsorption was not significantly affected by the temperature between 10 and 50 ℃. The X- ray diffraction analysis indicated occurrence of the intercalation of the rectorite by the toxin. The infrared absorption spectrum showed that the binding of the toxin did not alter its structure. The LC50 wlues of the adsorbed toxin were smaller than those of the free toxin. The rectorite protected the toxin from ultraviolet irradiation damage. The desorption of the adsorbed toxin in water ranged from 37.5% to 56.4% and from 27.4% to 41.8% in a carbonate buffer. The desorption percentage also decreased with increasing rectorite：toxin ratio.
基金supported by the National Key Research and Development Program of China(2018YFD0900505 to Y.-A.Z.and 2018YFA0801000 to Y.S.)the National Natural Science Foundation of China(32025037 and 31721005 to Y.S.)。
文摘Suppressive regulatory T cells(Treg cells)play a vital role in preventing autoimmunity and restraining excessive immune response to both self-and non-self-antigens.Studies on humans and mice show that the Forkhead box p3(Foxp3)is a key regulatory gene for the development and function of Treg cells.In zebrafish,Treg cells have been identified by using foxp3a as a reliable marker.However,little is known about the function of foxp3a and Treg cells in gonadal development and sex differentiation.Here,we show that foxp3a is essential for maintaining immune homeostasis in zebrafish testis development.We found that foxp3a was specifically expressed in a subset of T cells in zebrafish testis,while knockout of foxp3a led to deficiency of foxp3a-positive Treg cells in the testis.More than 80%of foxp3a^(-/-)mutants developed as subfertile males,and the rest of the mutants developed as fertile females with decreased ovulation.Further study revealed that foxp3a^(-/-)mutants had a delayed juvenile ovary-to-testis transition in definite males and sex reversal in about half of the definite females,which led to a dominance of later male development.Owing to the absence of foxp3a-positive Treg cells in the differentiating testis of foxp3a^(-/-)mutants,abundant T cells and macrophages expand to disrupt an immunosuppressive milieu,resulting in defective development of germ cells and gonadal somatic cells and leading to development of infertile males.Therefore,our study reveals that foxp3a-positive Treg cells play an essential role in the orchestration of gonadal development and sex differentiation in zebrafish.
基金supported by the project from China Ocean Mineral Resources R&D Association(COMRADY135-B2-17)+1 种基金the National Key Technologies R&D Program of China(2018YFD0500203 and 2018YFF0213503)the Major Projects of Technological Innovation in Hubei Province,China(2017ABA072)。
文摘Root-knot nematodes(RKNs) cause huge yield losses to agricultural crops worldwide. Meanwhile, livestock manure is often improperly managed by farmers, which leads to serious environmental pollution. To resolve these two problems, this study developed a procedure for the conversion of chicken manure to organic fertilizer by larvae of Hermetia illucens L. and Bacillus subtilis BSF-CL. Chicken manure organic fertilizer was then mixed thoroughly with Paenibacillus polymyxa KM2501-1 to a final concentration of 1.5×10^(8)CFU g^(-1). The efficacy of KM2501-1 microbial organic fertilizer in controlling root-knot nematodes was evaluated in pot and field experiments. In pot experiments, applying KM2501-1 microbial organic fertilizer either as a base fertilizer or as a fumigant at the dose of 40 g/pot suppressed root-knot disease by 61.76 and 69.05% compared to the corresponding control treatments, respectively. When applied as a fumigant at the dose of 1 kg m;in field experiments, KM2501-1 microbial organic fertilizer enhanced the growth of tomato plants, suppressed root-knot disease by 49.97%, and reduced second stage juveniles of RKN in soil by 88.68%. KM2501-1 microbial organic fertilizer controlled RKNs better than commercial bio-organic fertilizer in both pot and field experiments. These results demonstrate that this co-conversion process efficiently transforms chicken manure into high value-added larvae biomass and KM2501-1 microbial organic fertilizer with potential application as a novel nematode control agent.
