Meloidogyne spp., are globally emerging harmful animals for many agricultural crops.The root-knot nematodes (RKN), especially Luffa cylindrica) infected and non-infected by M. incognita nematodes, were investigated using culture-dependent and -independent methods.To explore microbial agents for biological control of these nematodes, the microbial communities of the rhizosphere soils and roots of sponge gourd  were shared, with Arthrobacter sp. and Bacillus sp. being dominant. Pseudomonas nitroreducens was exclusively present in the infested soils, while Mammaliicoccus sciuri, Microbacterium azadirachtae, and Priestia sp., together with Mucor irregularis, Penicillium sp., P. commune, and Sordariomycetes sp. were found only in the uninfected soils. Cupriavidus metallidurans, Gordonia sp., Streptomyces viridobrunneus, and Terribacillus sp. were only in the uninfected roots while Aspergillus sp. only in infected roots. After M. incognita infestation, 319 bacterial OTUs (such as Chryseobacterium) and 171 fungal OTUs (such as Spizellomyces) were increased in rhizosphere soils, while 181 bacterial OTUs (such as Pasteuria) and 166 fungal OTUs  rose their abundance in plant roots. Meanwhile, much more decreased bacterial or fungal OTUs were identified from rhizosphere soils rather than from plant roots, exhibiting the protective effects of host plant on endophytes. Among the detected bacterial isolates, Streptomyces sp. TR27 was discovered to exhibit nematocidal activity, and B. amyloliquefaciens, Bacillus sp. P35, and M. azadirachtae to show repellent potentials for the second stage M. incognita juveniles, which can be used to develop RKN bio-control agents.Thirty-two culturable bacterial and eight fungal species, along with 10,561 bacterial and 2,427 fungal operational taxonomic units (OTUs), were identified. Nine culturable bacterial species, 955 bacterial and 701 fungal OTUs were shared in both four groups. More culturable bacterial and fungal isolates were detected from the uninfected soils and roots than from the infected soils and roots (except no fungi detected from the uninfected roots), and among all samples, nine bacterial species (These findings provided insights into the interactions among root-knot nematodes, host plants, and microorganisms, which will inspire explorations of novel nematicides. RKN infDifferent methods have been employed to control these vital RKNs in integrated pest management programs, such as cultural practices, resistant crop cultivars, chemical nematicides, and biocontrol . HoweverBacillus firmus YBf-10 efficiently reduces damage of M. incognita to tomato plants with reduced galls, egg masses on roots, and nematode counts in soil  and ITS (for fungi) sequences. The microbiomes of infested and non-infested rhizosphere and soils and roots were used to examine their biological potentials for the pre-J2s of M. incognita.It is known that the soil conditions or plant species influence the bacterial and fungal community . Both in2.2.1.L. cylindrica were removed from the soil to observe the severity of root symptoms. Many galls were present in M. incognita-infested plant roots, while non-infested roots had no galls . Whole plants of no galls . The roono galls . Five reMeloidogyne incognita was collected from the sponge gourd roots in Baiyun District, Guangzhou, China  for the bioassay below and identified by SCAR marker (R marker . The pla2.2.−1 g/mL). Meanwhile, 5 g of the sterilized plant roots from each sample were ground with a sterile mortar, mixed with 45 mL sterile PBS, and also labeled as a start concentration (10−1 g/mL). The sample suspension was used to isolate bacteria and fungi on the plates of Trypticase soy agar , Potato dextrose agar (PDA), and Gause’s synthetic agar . The resulting plates were sealed with parafilm  and cultured in the dark at 26°C. Nine plates were set up for each sample and each dilution level . After 3 days, the bacterial and fungal colonies appeared on the plates were individually collected and inoculated into new plates. Microbe isolates were first grouped according to their colony and morphological characteristics. The resulting colonies were purified on the new plates and cultured in liquid LB  or PDA (for fungi) and kept at −80°C with 15% sterile glycerol. The DNA from each isolate was extracted using HiPure Bacterial DNA Kit and Fungal DNA Kit . The prokaryotic V4 region of bacterial 16S rRNA was amplified with primers 27F/1492R, and the ITS2 region of fungal ITS gene was amplified with ITS1F/ITS4R  according to the manufacturer’s instructions. After quantification by the NanoDrop ND-3300 spectrophotometer , PCR was carried out to generate amplicons in quintuplicate for rhizosphere soil samples and quadruplicate for plant root samples. The V5–V6 region of the bacterial 16S rRNA gene was amplified using primers 799F/1193R, and the ITS1 region of the fungal ITS gene was amplified using the primers BD-ITS1F/ITS2-2043R . All thehttps://github.com/vegandevs/vegan/) and Mothur software . A linear discriminant analysis (LDA) effect size (LEfSe) algorithm (The OTUs were grouped with a threshold of 97% pairwise identity by Usearch software (v. 10.0.240). The most frequently occurring sequence was extracted as the representative sequence for each OTU and used for taxonomic annotation. The taxonomic classification was performed by SILVA v. 119;  and UNITlgorithm  was empl2.4.Thirty-three representative bacterial isolates from the culture-dependent communities were selected from the RKN-infested and uninfested groups for nematode dispersal assay . The bac2.5.The nematicidal activity of microbial isolates from culture-dependent microbial communities , was det2.6.t-test with a threshold of p < 0.05.Statistical analysis was carried out by GraphPad Prism 5.0 software program . Significant differences in nematicidal activity and nematode dispersal assays were determined by one-way ANOVA or Student’s 3.3.1.Arthrobacter sp., Bacillus sp., B. ambifaria, Enterobacteriaceae sp., F. barbaricus, Microbacterium sp., Micrococcaceae sp., Rhizobiaceae sp., and Serratia sp.) were shared, and Arthrobacter sp. and Bacillus sp. were dominant species, among all samples. On the contrary, no fungal species overlapped among the samples.Overall, 32 bacterial species belonging to 23 genera from five classes, and eight fungal species belonging to five genera from five classes were identified from the soils and roots . At the M. incognita infested samples, more bacterial colonies were detected from the uninfested samples, including root and soil samples  were detected only from soils (US and IS). In comparison, seven root-intrinsic (UR and IR) bacterial species were identified, including Gordonia sp., C. metallidurans, Paenibacillus sp., P. glycanilyticus, Sporosarcina koreensis, S. viridobrunneus, and Terribacillus sp. Compared with the uninfested soils, more colonies of Chryseobacterium sp., E. adhaerens, Flavobacterium sp., Mesorhizobium sp., and S. puteale, and litter colonies of Comamonas sediminis, F. barbaricus, and Serratia sp. were detected from the infested soils  and 1,540,655 fungal (ITS) clean reads were obtained from 18 samples representing rhizosphere soils  and plant roots . Each sample with over 96% of reads met the demand of Q30 . A totalRalstonia sp., Bacillus sp., Sphingomonas sp. from bacteria, and Thermomyces sp., Mortierella alpina, Penicillium simplicissimum, and Orbilia aurantiorubra from fungi. The unique bacterial and fungal OTUs were detected in all groups but showed very low abundances. Five hundred and five bacterial and 154 fungal OTUs were shared between US, IS, and IR groups, suggesting these inherent soil OTUs may spread to the roots during M. incognita infestation. In contrast, the M. incognita infestation may also mediate the inherent root OTUs translocated to the soils, including 19 bacterial and 37 fungal OTUs shared in UR, IR, and IS groups. Furthermore, 57 bacterial and 35 fungal OTUs were specifically present in IS and IR. These results suggested that M. incognita may mediate the microbial exchange between rhizosphere soils and plant roots. On the other hand, 29 bacterial and 53 fungal OTUs vanished during the infestation, demonstrating the negative effect of M. incognita on microorganism communities  was much more than in plant roots , and similar results were found in fungi  and IS , while there was no significant difference in fungal infested groups  and Simpson  index of fungi in US were significantly higher than in UR, together with the rank-abundance results, showing more microbes species but less evenness in the uninfested soils than that in the uninfected roots. In addition, a similar Chao1 index  but a significantly higher Simpson index  in the fungal groups were detected in IS, compared with the US, suggesting that the infestation of M. incognita may decrease the fungal diversity in the soils.Compared with UR and IR samples for the alpha-diversity analysis, significantly higher bacterial and fungal abundance were detected in US and IS, as indicated by the Chao1 index . The Sim2R = 0.631, p = 0.001, 2R = 0.361, p = 0.006) and two root samples , but no significant difference was observed between UR and IR samples , OTU_76 (Flavobacterium sp.), and OTU_4 (Pseudomonas sp.) was significantly increased, while OTU_8 (Chujaibacter soil) and OTU_35 (Bacillus sp.) were decreased. In plant root samples, infestation with M. incognita significantly increased OTU_6 (Burkholderiaceae sp.). The fungal beta-diversities also clustered the samples into the same two parts as bacterial ones. The top 20 most abundant rhizosphere fungal OTUs accounted for more than 73% of total fungal abundance. However, the top 20 most abundant plant root endophytic fungal OTUs represented less than 15%, indicating the unevenness of the fungal communities in rhizosphere soils. Moreover, the infestation of M. incognita could significantly increase the abundance of OTU_2 (Fusarium sp.) and OTU_22  in the soil samples.In addition, UPGMA clustering analysis was also done to evaluate the beta-diversity changes in four groups . The bac3.3.M. incognita invasion were listed in M. incognita infestation. However, the decreased bacterial OTUs were only a quarter of the increased OTUs  in plant root samples. On the other hand, the number of decreased fungal OTUs was higher than that of increased in rhizosphere soils , while it was reversed in plant roots . This implied that the endophytic microbes in plant roots were not adversely influenced by nematode invasion.The bacterial and fungal OTUs that increased or decreased after Ralstonia sp.), was the dominant OTU shared among four groups. Three dominant bacterial OTUs were shared by US and IS, while UR and IR shared eight dominant bacterial OTUs. Moreover, for the rhizosphere soil groups, the abundance of OTU_31 (Myroides sp.), OTU_1957 (Flavobacterium sp.), OTU_76 (Flavobacterium sp.), OTU_18 (Chryseobacterium sp.), and OTU_10 (Chryseobacterium sp.) in IS was at least 15 times higher than that in the US. On the contrary, OTU_8 (Chujaibacter soli), OTU_23 (Bacillus sp.), and OTU_27 (Gammaproteobacteria sp.) in the IS were suppressed to less than a quarter after M. incognita infestation (Pasteuria sp.), and OTU_111 (uncultured bacterium) in IR significantly increased by at least 39 times compared with UR, while two Chitinophagaceae bacteria (OTU_150 and OTU_153) were dropped to less than a tenth. Specifically, the dominant OTU in UR, OTU_220 (Xanthobacter flavus), vanished after the M. incognita infestation  were identified from US, UR, IS, and IR groups, respectively . One bacestation . For plaestation .Fusarium sp.) was shared among four groups, especially with high relative abundance in soil samples . Moreover, 10 dominant OTUs were shared in rhizosphere soils, and 13 dominant OTUs were shared in plant roots. For soil groups, seven OTUs significantly increased their abundance when M. incognita infestation. OTU_22  was 151 times more abundant in IS than in US groups. On the contrary, OTU_23 (Trichoderma sp.) decreased to a fifth in IS compared to US groups. For root groups, the abundance of OTU_50 (Ceratobasidiaceae sp.), OTU_281 (Fungi sp.), and OTU_321 (O. aurantiorubra sp.) was significantly increased after infestation, while OTU_49 (Rhizoctonia sp.), OTU_3063 (Rhizoctonia sp.) and OTU_126 (Branch06) was suppressed to less than one fifth  and P. simplicissimum  shared between rhizosphere soil groups. Onygenales sp.  and Spizellomyces sp.  increased abundance in IS by more than 100 times compared with US, while Agaricomycetes sp.  was significantly suppressed to a twentieth in IS. On the other hand, Helotiaceae sp. , Pleosporales sp. , Alternaria sp. , and Malassezia arunalokei  existed in both root samples. After M. incognita infection, Exophiala sp.  increased 180 times higher in IR compared to UR, while Branch06 sp.  was significantly decreased  and plant roots (such as Pasteuria and Thiobacillus), respectively. Meanwhile, 56 and 15 genera declined their abundance in soils (such as Proteiniclasticum and Granulicella) and roots (such as Tistrella and Aquamicrobium). Particularly, the abundance of four genera was altered differently in rhizosphere soils and plant roots. Fluviicola and Peredibacter increased their abundance in rhizosphere soils but decreased in plant roots, whereas Deinococcus and Saccharopolyspora were reversed. For fungi, 13 (37) and 21 (13) genera were significantly increasing (decreasing) their abundance in rhizosphere soils and plant roots, respectively. Dactylella was specifically present in the infested rhizosphere soils, while the entomopathogenic fungi Beauveria bassiana was significantly increased in IS. Remarkably, except for Fusarium, 36 out of 37 decreased genera in rhizospheric soils were not changed in roots, such as Mortierella, Ascobolus, and Thielavia. Detailed information on other taxonomic levels is shown in In addition to the major taxa above, the other taxa from different taxonomic levels that significantly varied after so shown . For bac3.4.M. incognita infestation. A total of 52 bacterial taxa were identified as biomarkers (Logarithmic LDA > 4), including 22 from US, 10 from UR, 9 from IS, and 11 from IR. Exhibiting more discriminative taxa in uninfested rhizosphere soils than in infested soils. Genus Bacillus  was the most significant taxa in US, followed by Chujaibacter , while the genus Flavobacterium  had the largest effect sizes in IS. The most significant genus taxa in UR was Methylobacterium , and three genera with the largest effect sizes in IR were Geobacillus , Ralstonia , and Pasteuria  and Mortierella  from US, which are species P. simplicissimum  and M. alpina  belonged; Thermomyces  from IS; Leveillula  and Alternaria  from UR , while no bacterial isolates attracted nematodes  from the infested roots (not present in the uninfected roots) showed the strongest toxicity to M. incognita pre-J2s at 48 h post incubation  at 12, 36, and 48 h  and S. spiritivorum (2.79 times). Sphingobacterium psychroaquaticum has the antagonistic ability on fungi pathogen, which could elicit the induced systemic resistance of the host plant  and Pseudomonas bacteria in rhizosphere soils were 5.73 and 3.7 times higher after M. incognita infestation. It seemed that Flavobacterium and Pseudomonas bacteria in rhizosphere soils play special roles during M. incognita infestation, whether they have anti-nematode activity or plant growth ability need further research. On the contrary, genera Arthrobacter, Bacillus, and Chujaibacter decreased their abundance. Bacillus bacteria shows significant anti-nematode activity by producing plantazolicin (such as B. amyloliquefaciens FZB42) or inducing systemic resistance in plants (such as B. atrophaeus GBSC56) (Bacillus colonization was negatively correlated with pathogen abundance and disease incidence (Bacillus bacteria (US vs. IS: 3.82% vs. 1.43%) in rhizosphere soils significantly decreased after M. incognita infestation. Interestingly, two Bacillus species (B. amyloliquefaciens and Bacillus sp.) isolated by culture-dependent method repelled M. incognita pre-J2s. Why M. incognita presence decreased the abundance of Bacillus species which can repel the nematodes needs further investigation. In addition, M. azadirachtae, a plant growth-promoting actinobacterium (nt crops . Most of effects . In our  GBSC56) . Moreovencidence . Consistacterium , which rBacillus, Flavobacterium, and Pseudomonas resulted from the interaction among nematodes, fungi, and plants in rhizosphere soils. However, all of these bacterial genera were potential nematicides. Chujaibacter soli is essential in polyfluoroalkyl substances and lindane degradation, while Arthrobacter sp. can degrade unusual and polymeric compounds  is a filamentous fungus that can secrete a variety of extracellular enzymes and it is a well-known biological insecticide, including nematicidal activity  were suppressed in rhizosphere soils after M. incognita infestation. Species of Catenaria are known primarily as pathogens of nematodes  that plays a protective role from plant pathogenic infection. Some species of genera Geobacillus and Pasteuria in rhizosphere soils or plant roots show insecticidal or nematicidal activity, such as G. thermoglucosidasius and P. penetrans , but suppress the fungi with antibiotic activity , as well as increase the bacteria associate with reducing soils’ ability to recover from pollutions (such as Chujaibacter and Arthrobacter). Meanwhile, in response to the M. incognita invasion, the root exudates may provide a better environment to promote the growth of nematode eaters or plant growth promotors . On the other hand, in the infected plant roots, the host plant trended to protect the beneficial bacteria that were suppressed in the infested soil , and increased the abundance of insecticidal or nematicidal  and plant growth-promoting (Streptomyces and Methylobacterium) bacteria. And may be due to the enhancement of the immune system, toxin-secreting fungi  were eliminated in the infected plant roots. More candidate microorganisms are listed in In summary, in the case of 5.M. incognita, 32 bacterial and 8 fungal isolates were detected by culturable method. Ten thousand five hundred and sixty-one unculturable bacterial and 2,427 fungal OTUs were also obtained. Supernatant of Streptomyces sp. from the uninfected soils exhibited approximately 67.59% nematocidal activity against M. incognita pre-J2s. B. amyloliquefaciens, Bacillus sp., and M. azadirachtae repelled the nematodes. M. incognita invasion significantly influenced the microbiota composition in rhizosphere soils and plant roots. These results provided insights into exploring novel microbiological agents for safe control of these harmful plant nematodes.In this study, from the surrounding soils and roots of sponge gourd with and without The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found in the article/Meloidogyne incognita. LQ conducted the research and wrote the manuscript. LQ and ZR analyzed the data. RH and ZR revised the manuscript. All authors contributed to the article and approved the submitted version.RH designed and coordinated the research. LQ, LC, and JW collected the soil and root samples. LQ and KD collected the eggs and pre-J2s of The work was supported by the GDAS Special Project of Science and Technology Development (2022GDASZH-2022010101) and the Guangzhou Science and Technology Project (202206010120).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Aspergillus fumigatus and its potential translational application including noninvasive diagnosis of fungal infections via urine samples, imaging of fungal infections via labeling of siderophores with radionuclides such as Gallium-68 for detection with positron emission tomography, conjugation of siderophores with fluorescent probes, and development of novel antifungal strategies.Iron is an essential trace element that is limiting in most habitats including hosts for fungal pathogens. Siderophores are iron-chelators synthesized by most fungal species for high-affinity uptake and intracellular handling of iron. Moreover, virtually all fungal species including those lacking siderophore biosynthesis appear to be able to utilize siderophores produced by other species. Siderophore biosynthesis has been shown to be crucial for virulence of several fungal pathogens infecting animals and plants revealing induction of this iron acquisition system during virulence, which offers translational potential of this fungal-specific system. The present article summarizes the current knowledge on the fungal siderophore system with a focus on  Aspergillus fumigatus [3+)-specific chelators employed by bacteria and fungi for iron acquisition and by fungi also for intracellular iron handling [A. fumigatus, as SIA has been studied in most detail in this mold. A. fumigatus has been shown to employ RIA and SIA but appears to lack efficient use of heme as iron source [Saccharomyces cerevisiae and A. fumigatus; the transporter is termed CccA in A. fumigatus, whereby it is unknown if vacuolar stored iron can be reused in A. fumigatus [A. fumigatus are summarized in Fungi colonize an immense range of habitats involving mutualistic, competitive, and pathogenic relationships with other organisms . Due to umigatus . Siderophandling ,9. The pn source . Controlumigatus . The strAscomycota and Basidiomycota species produce hydroxamate-type siderophores displaying a remarkable species-specific structural variety [Saccharomycotina clade, including Saccharomyces cerevisiae and Candida albicans, as well as the basidiomycete genus Cryptococcus spp. [Mucoromycota lack hydroxmate-type siderophores but produce a carboxylate-type siderophore, termed rhizoferrin, originally isolated from Rhizopus microsporus [Rhizopus delemar [ferrichrome) and in some cases to the metal-free form . Siderophores are synthesized in the metal-free form but taken up only as metal complex. Detailed chemistry of the structural diversity of fungal siderophores has been reviewed previously [Most  variety . Howevercus spp. . Mucoromrosporus ,13. Rhizrosporus . Rhizofe delemar . Apart feviously ,15. In aeviously ,16. Planeviously .A. fumigatus  possess SITs [Uptake of hydroxamate-type siderophore-iron chelates in fungi is mediated by members of the siderophore iron transporter (SIT) family, which is a subfamily of the major facilitator superfamily. SITs are exclusively found in the fungal kingdom. Most species from all fungal phyla , Arn1p for ferrichromes, Taf1p/Arn2p for TAFC, Enb1p/Arn4p for the bacterial catecholate-type siderophore enterobactin [C. albicans contains only a single broad-range substrate-specific SIT, CaArn1p/CaSit1p [Saccharomycotina SITs are more similar to each other than they are to SITs from other fungal species [non-Saccharomycotina SITs, such as those of mold species, cannot be predicted on the basis of sequence similarity to S. cerevisiae SITs.Functional characterization of SITs started in robactin . In cont/CaSit1p . Phyloge species ,30, whicA. fumigatus can utilize a variety of hydroxamate-type siderophores, including several ferrichromes, endogenously secreted FsC and TAFC, several ferrioxamines, and coprogens, although the latter are utilized only poorly [A. fumigatus was found to be unable to utilize the hydroxamate-type siderophores basidiochrome and rhodotorulic acid, the bacterial catecholate-type siderophore enterobactin, rhizoferrin, and the mixed-type siderophores ornibactin and schizokinen [A. fumigatus [1 and R2 and acetyl as acyl-group in R4–R6; (ii) Sit2 but not Sit1 accepts anhydromevalonyl as acyl-group in positions R4–R6; (iii) Sit2 does not distinguish between cis- and trans-anhydromevalonyl as acyl-group in positions R4–R6; (iv) Sit1, and to a lesser extent Sit2, accept methylglutaconyl as acyl-group in positions R4–R6; (v) methylglutaconyl as acyl-group in R4–R6 significantly decreases uptake efficacy in comparison with anhydromevalonyl; and (vi) Sit1 accepts asparagine, leucine, and D-phenylalanine in positions R1, R2, and R3, while at least one of these amino acid residues disturbs recognition by Sit2 and therefore uptake of the ferrichrome-type antifungal VL-2397 depends exclusively on Sit1 [1–R3 as well as the acyl-groups in positions R4–R6 impact recognition of ferrichrome-type siderophores. Similar to ferrichrome A, the utilization efficacy of coprogen-type siderophores was low despite the fact that these siderophores were accepted by both Sit1 and Sit2 [A. fumigatus, a corresponding major transporter has been identified. Sit1, Sit2, and MirD were found to be dispensable for virulence in murine models of pulmonary aspergillosis [Aspergillus nidulans SIT MirA was indicated to transport bacterial catecholate-type siderophore enterobactin [y poorly . In contizokinen . Recent umigatus ,32,33. Sumigatus , which i on Sit1 ,34. Thesand Sit2 . Furtherand Sit2 , and thaand Sit2 . Acquisigillosis ,32,33. Irobactin . The insA. fumigatus SITs belong to different subclades [Saccharomycotina species S. cerevisiae, C. albicans, and C. glabrata are closely related building a sister clade with the ‘Sit1 clade,’ which indicates a common origin. Within the A. fumigatus Sit1 subclade, Fusarium graminearum Sit1 has been shown to transport ferrichrome and ferrioxamine B and Cryptococcus neoformans Sit1 was found to transport ferrioxamine B [Fusarium oxysporum, Aspergillus lentulus and A. nidulans. Importantly, the A. nidulans MirB homolog was shown to transport TAFC when heterologously expressed in S. cerevisiae [A. fumigatus, MirC, which might play a function in FC biosynthesis [S. cerevisiae Gex2 and Schizosaccharomyces pombe Str3 have been reported to transport glutathione and heme, respectively [S. pombe Str1 was found to transport ferrichrome [Saccharomycotina subclade.Phylogenetic analysis of 38 SITs from 13 fungal species shown in ubclades . As mentxamine B ,38, whicxamine B . Among trevisiae . Anotherynthesis . Notablyectively ,41. In crichrome . Taken tA. fumigatus EstB for TAFC, A. fumigatus SidJ for FsC, and A. nidulans EstA for enterobactin [Due to the high affinity of siderophores to iron, intracellular mechanisms to release the chelated iron from siderophores are required. Three esterases have been identified, which intracellularly hydrolyze siderophores with narrow substrate specificity: robactin . In bactrobactin . Such a A. fumigatus employs two iron-sensing transcription factors, termed SreA and HapX [2Cys2 GATA-type zinc fingers, which recognize the consensus DNA sequence ATCWGATAA, separated by a cysteine-rich region (CRR), which most likely mediates iron sensing. HapX comprises several phylogenetically conserved domains: (i) a bZIP-type DNA-binding domain, (ii) a Hap4-like domain (Hap4L) for physical interaction with the CCAAT-binding complex , and (iii) four CRR, whereby two CRR (CRR-A and CRR-B) have been implicated in iron sensing. During iron sufficiency, SreA transcriptionally represses high-affinity iron uptake, including RIA and SIA. During iron starvation, HapX represses iron-consuming pathways such as respiration, heme biosynthesis, TCA cycle, and vacuolar iron deposition to spare iron. Furthermore, HapX transcriptionally activates SIA [Ascomycota and Basidiomycota species [Saccharomycetaceae, including S. cerevisae but not Candida albicans, lost SreA, conserved only the iron detoxification function of HapX leading to Yap5 proteins, and evolved novel iron regulators termed Aft1/2 [A. fumigatus was shown to depend on mitochondrial but not cytosolic iron sulfur cluster biosynthesis [Similar to other high-affinity iron acquisition mechanisms, SIA is induced by iron limitation and repressed by iron to avoid excessive iron uptake. and HapX ,47. SreAates SIA . Remarkaates SIA ,49. Both species ,47. Inteynthesis . Most liynthesis .2Cys6-type transcription factor LeuB for feedback activation of leucine biosynthesis [A. fumigatus indicating integration of metabolic signals in control of iron homeostasis [Recent studies indicated that iron shortage is additionally sensed via iron-dependent metabolic pathways. Biosynthesis of the branched-chain amino acids leucine, isoleucine, and valine includes enzymes with iron–sulfur clusters as cofactors and consequently iron limitation leads to accumulation of the pathway intermediate α-isopropylmalate, which post-translationally activates the Znynthesis . Recentleostasis ,53. Moreeostasis ,55. Preveostasis ,57. Conseostasis ,58.in vitro growth of A. fumigatus in serum from hematopoietic stem cell-transplanted patients [Mucoromycota species in dialysis patients and the use of desferrioxamine against aluminum overload and/or iron excess, a link that was confirmed in animal models in 1989 [Ustilago maydis in 1993 largely diminished the interest in fungal SIA [A. fumigatus avirulent in murine models for invasive aspergillosis in 2004, while blocking RIA had no effect [F. graminearum, C. heterostrophus, and Alternaria brassicicola [U. maydis is a biotrophic pathogen that establishes a long-term feeding relationship with its hosts without causing immediate damage. Hemibiotrophs switch after a short initial biotrophic phase to necrotrophy. In the hemibiotrophic plant pathogen Colletotrichum graminicola, SIA was found to be repressed during biotrophic- and activated during necrotrophic growth [C. graminicola but only RIA for virulence of U. maydis [Mucoromycota species Mucor lusitanicus [Iron is an essential trace element that is limiting in most habitats including plant and animal hosts of fungal pathogens. Moreover, mammalian innate immunity further increases restriction of iron availability to fight infections within ‘nutritional immunity,’ leading to anemia of inflammation ,60. Conspatients . The fir in 1989 . Later o in 1989 . Howeverngal SIA . This vio effect . Subsequo effect ,33. Sidesicicola ,67–69. Tc growth . In line. maydis . The pre. maydis . Recentlitanicus .A. fumigatus, metabolic pathways that are required in a nonexclusive manner for SIA were found to be crucial for virulence of A. fumigatus including mitochondrial production of the siderophore precursor ornithine [E. festucae and its perennial ryegrass host [In line with the importance of SIA for virulence of rnithine , leucinernithine , biosyntrnithine , and biornithine . Lack ofass host ,78, whicA. nidulans [Escherichia coli or Salmonella spp. [Saccharomycotina or C. neoformans might be explained by the potential antagonistic activity of siderophores or alternatively to save energy as siderophore-chelated iron represents a soluble energized iron-form. The interplay between A. fumigatus and the bacterium Pseudomonas aeruginosa, which frequently coinfects the airways of cystic fibrosis patients, visualizes the antagonistic activity of siderophores: P. aeruginosa can iron-starve A. fumigatus via iron sequestration by its major siderophore pyoverdine, which cannot be utilized by A. fumigatus [A. fumigatus plays an important role in protection against P. aeruginosa [Pilobolus spp., which requires coprogen or ferrichrome as growth factor [Debaryomyces mycophilus, which lives as endosymbiont in the guts of woodlice, and the basidiomycete Tritirachium egenum, which is a mycosymbiont lacking independent high-affinity iron acquisition and depending on xenosiderophore supply during growth in association with Penicillium rugulosum [The battle for iron significantly also impacts the interaction of microorganisms leading to beneficial and antagonistic relationships. What is the reason for the stunning structural diversity of siderophores produced by fungi and bacteria? Most likely, the rational is that siderophores produced by a species can either promote or repress the growth of another species in the same habitat depending on its siderophore utilization capacity: growth promotion by improving iron uptake in case of uptake of the siderophore and growth repression by chelation of iron by a siderophore that is not recognized. The optimal competitive strategy would therefore be the production of siderophores that are not recognized by competitors combined with the capacity to utilize as many siderophore structures as possible. This is the most likely explanation for the utilization of xenosiderophores also by siderophore-producing species, e.g., utilization of enterobactin via MirA by nidulans  or the ulla spp. . Similarumigatus . In thisruginosa . In thisruginosa . The exih factor , the ascugulosum ,87.89Zr) for Positron Emission Tomography (PET), so-called immuno-PET imaging [in vivo release of 89Zr from the desferrioxamine B chelator has stimulated research to develop more stable chelators for PET imaging applications [68Ga) and 89Zr labeling [68Ga-labeled probes for hybrid imaging [89Zr revealed advantages over desferrioxamine B [The use of siderophores distinguishes microbial from plant and mammalian cells, which might enable translational applications. The most prominent translational application of siderophores is the bacterial siderophore desferrioxamine B , which has been used in clinics for metal chelation therapy in iron or aluminum overload pathologies . However imaging . The repications . Among tlabeling , as well imaging  and pret imaging  applicatxamine B .A. fumigatus is the most common human mold pathogen [A. fumigatus infection enabling noninvasive diagnosis in human urine and bronchoalveolar lavage [A. fumigatus rat infection model not only TAFC but also FC was detected in urine and FC also in serum [pathogen . Invasivpathogen . The diar lavage ,100. Detin serum , which ain serum . Howeverin serum ,104.68Ga allowed in vivo imaging of A. fumigatus infection by PET due to the specific uptake and accumulation of the siderophore in fungal cells [68Ga–desferrioxamine B for imaging pulmonary aspergillosis in a rat model [68Ga-labeling and PET [Acremonium persicinum and containing aluminum instead of iron, was recently shown to require the SIT Sit1 for uptake and activity against A. fumigatus, while its mode of action remains elusive [Furthermore, replacing iron in siderophores such as TAFC, ferrioxamine E, and ferrioxamine B by the radionuclide al cells ,105,106,al cells . Figure at model . Moreoveat model . Remarkaat model . These rat model ,111, in at model . Synthes and PET . The nat elusive . Notably elusive .Due to its crucial role in virulence, the siderophore biosynthetic pathway represents a potential target for selective therapeutic intervention. A natural quinone methide, celastrol, was identified as a noncompetitive inhibitor of SidA . MoreoveSiderophores play a central role in maintenance of iron homeostasis in most fungal species.Siderophores display a stunning species-specific structural variety.Siderophores play a crucial role mutualistic and antagonistic biotic interactions including fungal virulence in animal and plant hosts.Siderophores and siderophore uptake show great potential as biomarkers and for imaging of fungal infections.The siderophore system might allow fungal-specific drug delivery by a Trojan horse approach in which toxic compounds are conjugated to siderophores for selective import by fungal cells.


Vibrio vulnificus is an opportunistic, global pathogen that naturally inhabits sea water and is responsible for most vibriosis-related deaths. We investigated the genetic characteristics of V. vulnificus isolated from the clinical blood culture specimen of a patient with hepatitis B virus cirrhosis in 2018 (named as V. vulnificus VV2018) by whole genome sequencing (WGS). VV2018 belonged to a novel sequencing type 620 (ST620) and comprised two circular chromosomes, containing 4,389 potential coding sequences (CDSs) and 152 RNA genes. The phylogenetic tree of single nucleotide polymorphisms (SNPs) using 26 representative genomes revealed that VV2108 grouped with two other V. vulnificus strains isolated from humans. The pan-genome of V. vulnificus was constructed using 26 representative genomes to elucidate their genetic diversity, evolutionary characteristics, and virulence and antibiotic resistance profiles. The pan-genome analysis revealed that VV2018 shared a total of 3,016 core genes (≥99% presence), including 115 core virulence factors (VFs) and 5 core antibiotic resistance-related genes, and 309 soft core genes (≥95 and <99% presence) with 25 other V. vulnificus strains. The varG gene might account for the cefazolin resistance, and comparative analysis of the genetic context of varG revealed that two genes upstream and downstream of varG were conserved. The glycosylation (pgl) like genes were found in VV2018 compared with Pgl-related proteins in Neisseria that might affect the adherence of the strain in hosts. The comparative analysis of VV2018 would contribute to a better understanding of the virulence and antibiotic resistance profiles of V. vulnificus. Meanwhile much work remains to be done to better understand the function of pgl-like genes in V. vulnificus. Vibrio vulnificus is a gram-negative, rod-shaped bacterium that is widely distributed throughout marine and brackish environments , in Nantong, Jiangsu Province, China . Drug susceptibility was determined according to the Clinical and Laboratory Standards Institute (CLSI) drug susceptibility test standard from 2018.The antibiotic resistance profiles were assessed through minimal inhibitory concentration (MIC) assays . BrieflyV. vulnificus strains we used were further evaluated by checkM with default settings  and assembly were conducted at Azenta Life Sciences . Sequences of VV2018 were obtained using PacBio Sequel platform  and Illumina HiSeq X Ten platform . The PacBio reads were assembled by Hifiasm v0.13-r308 and Canu v2.2 , and thesettings . Prokka settings . The funsettings  against settings . The rRNsettings  and tRNAsettings , respectsettings . Genomicsettings . Multiloson Tool .V. vulnificus complete genome sequences (n = 25) were downloaded from the NCBI database with checkM values  and OmpU have been shown to play important roles in the infection and pathogenesis, respectively, of V. vulnificus  and Chr II . These genes were mainly associated with motility (polar flagellar proteins), immune evasion (capsular polysaccharide and iron uptake), secretion system (type II secretion system proteins), adherence  and toxin (RTX toxin) . Iron uplnificus . RTX toxlnificus . The vvhbacteria . In thisbacteria .dfrA3 (encoding a dihydrofolate reductase), qnrVC1 (encoding a pentapeptide repeat protein), catB9 (encoding a type B-5 chloramphenicol O-acetyltransferase), tet (crp (encoding a cAMP-receptor protein) and varG . CRP is a global regulator that not only regulates the expression of the multidrug efflux pump but also impacts the expression of multiple VFs , tet   and two tyrosine recombinases (xerC and xerD) were encoded. Two genes were predicted to encode type I restriction enzyme proteins. The length of GIs on Chr II ranged from 4 kbp to 86 kbp, containing one tyrosine recombinase-encoding genes (xerC). Meanwhile, sulfate permease genes (cysTWA) which allowed the bacteria survive in selenite environment by decreasing the expression, were found on Chr II_GI2 (Large parts of the genome designated as genomic islands (GIs) and phages were transferred from one bacterium to another . Twenty r II_GI2 . Only onr II_GI2 . Thus, fCas genes on Chr I  (V. vulnificus FORC_017 (98.45% identity) isolated from human in South Korea. The heatmap showed that all 26 V. vulnificus strains were divided into two clusters, most strains including VV2018 in cluster 1 were isolated from human, and most strains in cluster 2 were isolated from seafood or unknown places. Meanwhile, the phylogenetic tree of all 26 V. vulnificus strains constructed on the basis of the core SNPs showed that VV2018 was grouped with two other V. vulnificus strains isolated from human (V. vulnificus FORC_009 and V. vulnificus FORC_016) . A previous study reported that the value of 70% dDDH could not be used as absolute boundary, but still a gap between 60 and 70% similarity seemed to embrace clear-cut clusters of organisms, given the large extent of diversity among prokaryotes  . The mosORC_016) . The dDDkaryotes .V. vulnificus strains according to the pan-genome analysis. A total of 138 strain-specific genes, accounting for 3.14%, were identified in VV2018. The functions of the majority of VV2018 specific genes (78.26%) were unknown, the other specific genes were involved in functional categories of replication/recombination/repair (7.97%) and cell wall/membrane/envelope biogenesis (4.36%)  .V. vulnificus strains and 115 VFs were included in core genes. The heatmap based on the presence and absence of all VFs clearly showed that the distribution of virulence genes differed between V. vulnificus strains  were upstream and downstream of varG. The genetic context of varG was conserved and the genes upstream and down of varG were ompV and nodD that were present in 96.15% of the sequences. These observations indicated that sequence rearrangement rarely occurs in the varG-encoding region, and genetic commonalities of ompV-varG-nodD across sources strongly suggested the structure of ompV-varG-nodD was conserved in V. vulnificus strains.The genetic context of the resistance-related genes of VV2018 was almost the same compared with other 25 in vitro , howevery of 80% . The lary of 80% . The resNeisseria including N. elongata subsp. glycolytica, with 37% identity. Meanwhile, four genes downstream of LNNJENCE_00237 also shared a high identity compared with Pgl-related proteins in Neisseria such as N. elongata subsp. glycolytica ATCC29315 (pgl) systems have been defined in Neisseria, such as N. gonorrhoeae, N. meningitidis, and N. elongata subspecies glycolytica (pglA and pglBCD may be involved in pilin glycosylation (Two (LNNJENCE_00235 and LNNJENCE_00237) of six genes on Chr I_GI4 were glycosyl transferases, belonging to the GT4 family . LNNJENCTCC29315 . Broad-scolytica . The pglsylation .Neisseria for further analysis and the result of the multiple sequence alignment showed that the sequences of PglC and PglD were highly conserved  effectors by modulation of secretion efficacy (pgl-like genes (pglABCD) may play important roles in Vibrio species.We selected six sequences of onserved . The phyTCC29315 . MeanwhiFORC_017 . The res present . In V. cefficacy . TherefoNeisseria. The domains of PglA, PglBa, PglC and PglD were similar, except for PglBb  can be found in this article/The studies involving humans were approved by the Ethics Committee of Affiliated Nantong Hospital 3 of Nantong University. The studies were conducted in accordance with the local legislation and institutional requirements. The participants provided their written informed consent to participate in this study.TZ, QW, XuL, and XiL collected the strains and performed the experiments. MZ and TZ analyzed the experimental results. FW performed the bioinformatic analysis. FW, YZ, and RL co-led the writing of the manuscript. YZ and RL designed the work. All authors read and approved the final manuscript.


Akkermansia muciniphila, is considered the “next-generation probiotic” for alleviating metabolic disorders and the inflammatory response. Although extensive research has been conducted on A. muciniphila, none has summarized its regulation in T2D. Hence, this review provides an overview of the effects and multifaceted mechanisms of A. muciniphila on T2D and related diseases, including improving metabolism, alleviating inflammation, enhancing intestinal barrier function, and maintaining microbiota homeostasis. Furthermore, this review summarizes dietary strategies for increasing intestinal A. muciniphila abundance and effective gastrointestinal delivery.The prevalence of type 2 diabetes (T2D) is increasing worldwide, with many patients developing long-term complications that affect their cardiovascular, urinary, alimentary, and other systems. A growing body of literature has reported the crucial role of gut microbiota in metabolic diseases, one of which,  Diabetes mellitus (DM) refers to a group of metabolic disorders characterized by hyperglycemia, insulin resistance, obesity, and low-grade inflammation . The preRuminococcus, Fusobacterium, and Blautia) were enriched in T2D patients, while the genera Bifidobacterium, Bacteroides, Faecalibacterium, and A. muciniphila were negatively associated with T2D . A. muciniphila MucT (=ATCC BAA-835T = CIP 107961T), belonging to AmIa. Differences among these four phylogroups include intestinal abundance, physicochemical properties, metabolic characteristics, and immune activation capacity. In a study of 1617 human fecal samples  and glycosyltransferases (GTs), but the copy numbers of the GH and GT families differed from one another (12) is an essential cofactor for short-chain fatty acid metabolism. 12 synthesis genes were absent in AmI. This finding suggests that the vitamin B12 biosynthesis genes were originally present in all strains and were lost by some isolates in the human gut  were absent in AmII and AmIV. This resulted in the absence of hydrogen sulfide, a critical substrate for cysteine and methionine synthesis, and led to a low growth rate in the mucin medium  pathway analysis, genes in the tivities . Metabol another . The gen another . Notably another , and gal another . Similar another . Furtheruman gut . Consideubstrate , these gn medium . Addition medium . These rlogroups , and maylogroups .A. muciniphila and colitis is controversial. Several lines of evidence have shown that A. muciniphila administration can ameliorate inflammatory bowel diseases (IBD) or delay colitis-associated tumorigenesis in mice -10-deficient mice. Notably, A. muciniphila levels in MS patients. The investigators hypothesized that this difference might be due to strain specificity , and further investigation should be conducted to gain deeper insights into the interactions between the host and A. muciniphila.The relationship between  in mice , whereasiniphila . While sates IBD , Ring etcificity , and Liucificity . A. mucill lines . In summll lines , and A. A. muciniphila abundance in T2D patients as early as 10 years ago  is a pleiotropic hormone that has a broad role in metabolism regulation, including the stimulation of insulin secretion and appetite suppression , and indole derivatives. Propionate modulates the expression of genes involved in fatty acid uptake and oxidation, such as Fiaf, Gpr43, peroxisome proliferator-activated receptor gamma (PPARγ), and histone deacetylases (HDACs) (A. muciniphila administration reduces white adipose tissue (WAT) volume and enhances thermogenesis by upregulating the uncoupling protein 1 (Ucp1)-AMPK pathway and increases lipid oxidation, thereby ameliorating liver steatosis  . In mous 1 (Ucp1), and dow 1 (Ucp1). Increasd-change . Additioteatosis .A. muciniphila and quercetin, plasma primary bile acids to secondary bile acids ratio was significantly increased in mice  are steroid molecules that are synthesized from cholesterol in the liver and act as metabolic regulators via the nuclear farnesoid X receptor (FXR) and takeda G protein-coupled receptor 5 (TGR5). After t in mice . Besides in mice , which piniphila . The incth NAFLD ; therefoA. muciniphila promoted insulin secretion by limiting the availability of 3β-chenodeoxycholic acid (βCDCA) in mice. CDCA was shown to be a high-affinity FXR agonist synthesized in the liver  from the intestine into circulation. Notably, LPS infiltration exacerbated insulin resistance and diabetes and led to various diabetic complications . Improve studies . The intted mice , this mated mice . Both infunction .A. muciniphila, A. muciniphila increase the expression of tight junction proteins in mice, including zonula occludens-1 (ZO-1), occludin, claudin-3, junctional adhesion molecules-3 (jam-3)  proteins, with a scattered distribution of functionally specialized cells  . After t (jam-3) . These p (jam-3) . Howevernhibited . Notablyeability . ClaudinA. muciniphila supplementation induced variations in immune cell composition  is a transcriptional regulator widely expressed in the intestinal epithelium and mediates antimicrobial immunity. Furthermore, impaired AhR agonist activity and lower AhR ligand concentrations are associated with metabolic disorders and intestinal barrier dysfunction , and IL-22 levels that consequently alleviate metabolic syndrome and inflammation.The last line of defense consists of numerous immunocytes and immunoreactive substances. Several animal experiments have reported that position , and enhposition . The knotivation , or maintivation . Some antivation , α-tocoptivation , positivd leptin , displayfunction . AhR ligfunction . Gu et aA. muciniphila consolidates the intestinal barrier and regulates the balance of pro−/anti-inflammatory factors, thereby protecting the host from endotoxemia and metabolic disorders  and decreased levels of SCFA-producing bacteria   . The lowpatients . Althougnd Chao1 . These rFirmicutes and Bacteroidetes are the two most abundant phyla in the human intestine  play a significant role in improving the intestinal barrier  mediate signal transmission between the gut microbiota and the host . A. muci barrier , and reg barrier . Thus, A barrier . In summA. muciniphila on T2D and related metabolic diseases have been described previously, which remind us that A. muciniphila is a promising probiotic and that oral administration of A. muciniphila may alleviate metabolic disorders. The method of cultivation and administration of A. muciniphila can greatly affect its function. Owing to the potential of A. muciniphila in treating metabolic diseases, many researchers have explored the most suitable growing environment and method for A. muciniphila in the intestine. Suitable culture conditions will allow A. muciniphila to survive better so that they can be administered by gavage to observe their effect on metabolic diseases. When A. muciniphila was first discovered by A. muciniphila could grow under a low oxygen concentration , the Jilin Provincial Science and Technology Foundation (Nos. 20210509003RQ and 20210402002GH), and the Changchun Science and Technology Bureau Development Plan project (No. 21ZY29).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Hypsizygus marmoreus) is a highly nutritious, edible medicinal mushroom native to East Asia. The present research investigated the impact of different substrates on the metabolite compositions of H. marmoreus mycelia cultivated in vitro. The substrates tested included malt extract agar, malt extract agar enriched with barley malt, and malt extract agar enriched with grape pomace. The study also assessed antimicrobial and antiradical activities of the extracts against gram-positive bacteria (Bacillus subtilis and Staphylococcus aureus), gram-negative bacteria , yeasts , and dermatophytes . The results revealed that the H. marmoreus mycelia extracts demonstrated antibacterial and antifungal activities against the tested microorganisms. Extracts obtained from the cultivation in substrates enriched with either barley malt or grape pomace exhibited the highest antibacterial activity among all the tested bacterial strains except for P. aeruginosa. The same extracts showed the highest inhibitory effect against C. albicans and C. parapsilosis. Noteworthy, the extract from the mushroom cultivated in the substrate enriched with grape pomace also exhibited remarkable efficacy against T. mentagrophytes and T. tonsurans. Terpenoid and carbapenem compounds could be related to the antimicrobial properties of the extracts from mushrooms cultivated in substrates enriched with grape pomace. In comparison, the higher antiradical properties could be related to the content of indole compounds. In conclusion, growth substrate selection affects the nutritional and medicinal properties of H. marmoreus, making it a valuable contribution to the understanding of the cultivation of this mushroom.The beech mushroom ( The nutritional and health-promoting properties of mushrooms have been known for thousands of years. Ancient civilizations in China, Japan, and other eastern countries have long recognized the various biological activities of mushroom extracts ,2. EdiblHypsizygus marmoreus (Peck) H.E. Bigelow is well known for its nutritional and medicinal properties, among which are antitumor, antibacterial, antifungal, anti-inflammatory, antioxidant, antihypertensive, and antiallergic properties [H. marmoreus belongs to the Hypsizygus genus (Lyophyllaceae family) and includes four other species: H. elongatipes (Peck), H.E. Bigelow, H. ulmarius (Bull.) Redhead, H. tessulatus (Bull.) Singer, and H. ligustri Raithelh [operties ,8,9,10. Raithelh .H. marmoreus is considered a rare species.It can be found in different countries in North America, Asia, and Europe  . It thriH. marmoreus against eight phytopathogenic fungi, reporting significant inhibitory effects on Alternaria brassicicola. Shiono et al. [H. marmoreus, finding that one exhibited antifungal activity against the pathogenic fungus Raffaelea quercivora. Wong et al. [H. marmoreus, observing their inhibition effects on Candida albicans. The medicinal properties of H. marmoreus are attributed to several bioactive molecules, including terpenoid compounds like hypsiziprenol A9, which have been shown to exert cytotoxic effects in human liver cancer HepG2 cells [H. marmoreus are associated with various active components, mainly primary metabolites [H. marmoreus, known as a white-rot fungus [In addition to their well-known antioxidant activity, phenolic compounds found in mushrooms can exhibit significant antimicrobial effects against a wide range of organisms . For inso et al.  isolatedg et al.  evaluateG2 cells . The dieabolites . As a ret fungus , possesst fungus . Edible t fungus . Previout fungus . Solid ct fungus . Moreovet fungus . The intH. marmoreus mycelium based on preliminary experiments, (b) investigating and comparing the chemical profiles of H. marmoreus mycelium grown on these two different culture media, and (c) exploring the influence of medium ingredients on the antimicrobial and antiradical activities. To achieve these goals, we employed mass spectrometry (MS)-based metabolomics, which enables quantitative analyses with high selectivity and sensitivity, along with the potential to identify metabolites [H. marmoreus mycelia.In this study, our main objectives were as follows: (a) selecting two different solid media for cultivating abolites . CombiniH. marmoreus (fruiting body) was collected in Perugia (via Roma) in December 2018. The voucher specimen (PeruMyc2422) was deposited in the herbarium of the Department of Chemistry, Biology, and Biotechnology . The isolation of the mycelia was carried out as previously described [escribed .Genomic DNA was extracted from 15-day mycelia grown on malt extract agar (MEA) using the ZR Fungal/Bacterial DNA kit . The amplification of the internal transcribed spacer (ITS) region was conducted with the fungal-specific forward primer, ITS1F 5′-CTTGGTCATTTAGAAGTAA-3′ in combination with the reverse primer, ITS4 5′-TCCTCCGCTTATTGATATGC-3′ . The conH. marmoreus, Hypsizygus tessulatus (Bull.) Singer, and Hypsizygus ulmarius (Bull.) Redhead were retrieved from GenBank. Additionally, Ossicaulis lignatilis (Pers.) Redhead and Ginns was chosen as an outgroup for the ITS region analysis. Within the MEGA X package, the MUSCLE algorithm was utilized for pairwise and multiple sequence alignments, and the resulting alignments were further refined manually. To support the branches in the maximum likelihood (ML) analysis, a bootstrap (BS) value of 1000 pseudoreplicates was applied.The chromatogram was examined and edited using Chromas 2.6.6, while MEGA X software was employed for the subsequent sequence analysis. For the phylogenetic inference, the DNA sequences of H. marmoreus was carried out using the following solid media: (1) malt extract agar 1%, as the Hypsizygus control (HC); (2) malt extract agar 1% enriched with 0.5% barley malt (HC1); (3) malt extract agar 1% enriched with 0.5% grape pomace (HC2); (4) malt extract agar 1% enriched with 2% barley malt (HC3); (5) malt extract agar 1% enriched with 2% grape pomace (HC4). The details about the preparation of the samples are fully listed in our previous study [The in vitro cultivation of t (HC1);  malt extThe total phenolic contents of the tested extracts were evaluated via the colorimetric method using the method described for phenolics by Acquaviva et al. . The FolTo assess the antioxidant potential of the extracts, 2,2-Azino-bis-(3-ethyl-benzthiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) assays, were used to examine the antioxidants’ ability to neutralize free radicals . Each ofThe untargeted analysis was performed using a 1260 Infinity II LC System coupled with an Agilent 6530 Q-TOF spectrometer . The details are reported in our previous study [The antimicrobial properties of the extracts were investigated as previously reported ,28,29. FH. marmoreus and H. tessulatus . The acquired information included mass, retention time, and area of the five samples analyzed ,Regarding the metabolomics of The data produced in this experiment were explored using the statistical technique of unsupervised principal component analysis after applying autoscaling. The score plot graph  shows thThe dendrogram  shows thH. marmoreus were also tested for the amounts of phenolic compounds, calculated as gallic acid equivalents  exhibited the highest sensitivity to the H4 extract, with an MIC range of 1.56–3.125 μg mL−1 . In contrast, Bacillus cereus (PeruMycA 4) and Pseudomonas aeruginosa (ATCC 15442) showed the least sensitivity to the mycelia extracts. Generally, gram-negative bacteria  displayed lower sensitivity to the extracts compared to gram-positive strains, similar to observations for F. torulosa and Pleurotus spp. [−1) against Candida albicans (YEPGA 6379) and C. parapsilosis (YEPGA 6551), respectively (Trichophyton mantagrophytes (CCF 4823), T. tonsurans (CCF 4834), and T. rubrum (CCF 4933) being the most susceptible fungal species to all mycelia extracts, with MIC ranging from 6.26 to 200 μg mL−1. MIC values below 100 μg mL−1 were considered indicative of high antimicrobial activity , and medicinal plants , also in a context of sustainability through the use of by-products from a chain to feed the other one.In the present study, the influence of different substrates was investigated on the metabolomics profile and biological properties of extracts from the medicinal mushroom


Pseudomonas aeruginosa. Although incidence of CAP due to P. aeruginosa is low, it has been linked to severe illness and poor clinical outcomes, highlighting the challenge in treating this infection. Identifying local risk factors for pneumonia due to drug-resistant pathogens, specifically P. aeruginosa, can potentially reduce the overuse of unnecessary empiric broad-spectrum antipseudomonal agents and increase appropriateness of empiric antibiotic treatment regimens.Community-acquired pneumonia (CAP) is a leading infectious cause of hospitalization, morbidity, and mortality worldwide. As the etiology of CAP continues to evolve, antibiotic-resistant bacteria are becoming more prevalent in the community setting. Among these pathogens is P. aeruginosa group was composed of P. aeruginosa-positive respiratory specimens whereas the non-P. aeruginosa group comprised all other pathogens and specimen-negative results. The primary endpoint was to identify risk factors for P. aeruginosa in patients with CAP at our institution.A single-center, retrospective cohort study was conducted in patients who were treated for CAP at a large academic medical center from August 1, 2019 through July 31, 2022. Specimen data were reviewed and separated into two groups. The P. aeruginosa encounters and 40 non-P. aeruginosa encounters). Baseline characteristics were well-distributed between groups. A univariate analysis determined 13 variables were significant at P < 0.2. This P-value was used for entry into the logistic regression model. Of the 13 variables, four were determined to be correlated most closely with CAP due to P. aeruginosa: chronic pulmonary disease , prior intravenous antibiotics within the last 12 months , sputum production , and tube feeds at time of admission .A total of 206 patients were screened and 79 of them were included in the analysis (39 P. aeruginosa at our institution.Although the study sample size was small, chronic pulmonary disease, prior intravenous antibiotics within the last 12 months, sputum production, and tube feeds at time of admission correlated most closely with CAP due to All Authors: No reported disclosures


Acinetobacter baumannii (2009) in the United States, and no statewide prevalence survey has been performed for Candida auris. We aimed to determine the prevalence of A. baumannii and C. auris among mechanically ventilated patients in Maryland.To date, only one statewide prevalence survey has been performed for A. baumannii and antibiotic susceptibility testing was performed. Axilla/groin cultures were tested by PCR for C. auris.The Maryland MDRO Prevention Collaborative performed a statewide point prevalence survey of mechanically ventilated patients admitted to acute care hospitals (ACH) and long-term care (LTC) facilities between March 7, 2023, and June 8, 2023. Surveillance cultures were obtained from all mechanically ventilated patients. Sputum, peri-anal, and arm/leg cultures were cultured for A. baumannii was cultured from at least one body site in 148/482 (31%) patients, CRAB was identified in 88/482 (18%) patients and C. auris in 31/470 (7%) patients. LTC patients were more likely to be colonized with A. baumannii , with CRAB  and C. auris  compared to acute care hospital patients . Nine (29%) C. auris-positive patients were previously unknown to the Maryland Department of Health. Susceptibilities for A. baumannii are in Table 1.One hundred percent of all eligible healthcare facilities participated. 482 ventilated patients were included in the analysis. A. baumannii, CRAB and C. auris colonization are common among mechanically ventilated patients in both ACH and LTC facilities. Both pathogens were significantly more common in long-term care facilities than acute care facilities. Ventilated long-term care patients are an extremely high-risk population for emerging pathogens and surveillance and infection prevention efforts should be targeted to these facilities.Anthony Harris, MD, MPH, Merck: Grant/Research Support|UpToDate: Infection Control Editor


The innate system constitutes a first-line defence mechanism against pathogens. 80% of the blood supply entering the human liver arrives from the splanchnic circulation through the portal vein, so it is constantly exposed to immunologically active substances and pathogens from the gastrointestinal tract. Rapid neutralization of pathogens and toxins is an essential function of the liver, but so too is avoidance of harmful and unnecessary immune reactions. This delicate balance of reactivity and tolerance is orchestrated by a diverse repertoire of hepatic immune cells. In particular, the human liver is enriched in many innate immune cell subsets, including Kupffer cells (KCs), innate lymphoid cells (ILCs) like Natural Killer (NK) cells and ILC-like unconventional T cells – namely Natural Killer T cells (NKT), γδ T cells and Mucosal-associated Invariant T cells (MAIT). These cells reside in the liver in a memory-effector state, so they respond quickly to trigger appropriate responses. The contribution of aberrant innate immunity to inflammatory liver diseases is now being better understood. In particular, we are beginning to understand how specific innate immune subsets trigger chronic liver inflammation, which ultimately results in hepatic fibrosis. In this review, we consider the roles of specific innate immune cell subsets in early inflammation in human liver disease. Understanding the liver´s architecture and the niches formed by the different hepatic immune cells is equally important to deciphering their immune roles. The liver is subdivided into hepatic lobules, which consist of a portal triad , hepatocytes arranged in linear cords between a capillary network (sinusoids) and a central vein Figure 1+ monocytes participates in liver fibrosis in mouse models , DAMPs or cytokines alone. Upon activation, cells can execute cytotoxic as well as effector functions. Moreover, γδ T cells also play a role in tissue homeostasis  or microbial phosphoantigens , human immunodeficiency virus (HIV) and influenza  possess mechanisms to escape from the hosts´ antiviral immunity. When the viruses replicate, often the innate immunity detects viral components, hence triggering an acute inflammatory response resulting in the killing of infected hepatocytes. Since the infection is not properly resolved, viruses remain in a latent state and replicate opportunistically. This progressively leads to chronic liver inflammation . In partin vitro expression of TRAIL in hepatocytes , characterized by an excessive accumulation of fat in hepatocytes, is the most common indication for liver transplant in Western countries and the leading cause of liver transplantation in women , 132. Itatocytes . Immunohatocytes . Interesatocytes . Similaratocytes . Likewisatocytes  and impaatocytes , 145. Anatocytes . Data fratocytes –149. Adoia IL-10 . Howeveria IL-10 . This waia IL-10 . In the ironment . This coironment . Howeverironment . The hetironment  but incr4+ cells –158. NAS4+ cells . Taken t4+ cells . MAIT ce4+ cells . The cur4+ cells . γδ T ce4+ cells , a key c4+ cells , 163. Inelevated . Additioelevated , 161. Incontrols . Overallcontrols . Accumulcontrols . In linecontrols . KCs devcontrols .bright NK cells was higher in untreated AIH, while the frequency of circulating CD56dim NK cells was reported to be reduced in active AIH patients or while in remission , primary biliary cirrhosis (PBC) and primary sclerosing cholangitis (PSC). AIH affects portal tracts and liver lobules by lymphoplasmacytic infiltrates while PSC and PBC mainly affect bile ducts. The etiologies of these diseases are yet unknown, but several studies suggest a common immune-mediated liver injury. The dysregulation of immune regulatory networks causes the activation and expansion of autoreactive T cells and B cells , 170. Themission , 172. Ouemission . Activatecretion , 175. Inof IL-17 . MAIT ceof IL-17 . Chronicof IL-17 . The freof IL-17 . Vδ1+ T with AIH . Anotherwith AIH . TOX defwith AIH . In genewith AIH , 181.Liver transplantation represents a major hepatic injury. One of the unavoidable injuries is caused by oxygen deprivation. After liver resection, blood flow is restricted for a period of time and the organ becomes hypoxic. This leads to different forms of cell death like apoptosis, ferroptosis, pyroptosis and necrosis . After rin vitro  therapy, have opened new clinical perspectives for cancer patients and is fast becoming one of the main pillars of cancer treatment. ICI therapy uses monoclonal antibodies blocking T cell receptors that are used by cancer cells to evade the immune system. Immune-related adverse events (irAEs) are the result of immune activation derived from ICI therapy. The incidence of ICI-derived hepatitis is approximately 1-3% for programmed cell death 1 (PD1) inhibitors and 3-9% in cytotoxic T-lymphocyte-associated protein 4 (CTLA4) inhibitors . The comin vitro . The frein vitro . Overallin vitro . α-PD-1 in vitro .via three steps consisting of early inflammation, amplification of the inflammatory signal and resolution. Liver fibrosis is a consequence of inflammation and inefficient resolution. Liver biopsy is the gold standard for diagnosing cirrhotic liver disease, yet it is estimated to miss 10-30% of cases (The innate immune system is also involved in immune homeostasis and healthy tissue turnover. This is accomplished of cases . Additioof cases . An optiof cases . The curof cases . The immJYZ performed literature search and wrote the manuscript. The author confirms being the sole contributor of this work and has approved it for publication.


Smoking is one of the predictors of decreased cardiopulmonary endurance. Gait disturbance may be due to many reasons, including cardiovascular endurance. This study aimed to determine differences in gait parameters between non-smoker and smoker participants. A cross-sectional design was employed, involving thirty non-smokers and thirty-seven smokers as participants. Detailed interviews were conducted to gather information on smoking habits, status, and history. Gait parameters were measured using a high-quality 3D accelerometer, 3D gyroscope, and barometric pressure sensors (Physilog4 from GaitUp). Anthropometric characteristics were described, and mean values with standard deviations (SD) were calculated. An independent two-tailed t-test was conducted to compare gait parameters between non-smokers and smokers, with statistical significance set at p<0.05. The analysis revealed significant differences in various gait parameters between non-smokers and smokers. Specifically, significant differences were found in cadence , stride length , stride velocity , stance , swing , foot flat , pushing , and double support . However, no significant difference was found between non-smokers and smokers in the loading phase . There were significant differences in general and temporal gait parameters between smokers and non-smokers. Gait parameters provide valuable insights for evaluating functional performance and providing objective and quantitative data to assess gait disorders. Future studies should include longitudinal studies with large sample sizes to explore the effects of potential confounders on gait parameters. Smoking is one of the leading causes of mortality and morbidity. According to the Saudi Ministry of Health, smoking kills 71 men and 21 women each week, equating to 5,000 deaths per year due to smoking-related ailments. Smoking has been linked to various serious health conditions, including cancer, heart disease, stroke, chronic obstructive pulmonary disease (COPD), diabetes, and even erectile dysfunction in males ,4. VitalThe most prevalent human movement is gait, a voluntary and cyclical form of vertical mobility characterized by the alternating and progressive action of the lower limbs . Gait isCigarette smoking is a well-known and significant risk factor for various cardiovascular diseases, encompassing a range of disorders affecting the heart and blood vessels . SmokingAs a multidimensional phenomenon, gait can be thoroughly studied using various techniques. Gait analysis encompasses the exploration of multiple interacting characteristics with the aim of better understanding potential disruptions or limitations. The objective of this study was to compare the general and temporal gait parameters between non-smokers and smokers. The relationship between smoking and particular spatiotemporal metrics may indicate the processes underlying their relationships with gait. This knowledge can lead to the identification of improved intervention options to prevent future gait disturbances in smokers and non-smokers. Such information may be valuable when considering rehabilitation strategies or making complex therapeutic decisions.This cross-sectional study was conducted between May 2022 and October 2022 at the Department of Physical Therapy, Imam Abdulrahman Bin Faisal University, located in Dammam, Kingdom of Saudi Arabia.The sample size was calculated using G*Power (3.1.9.4) software. The parameters used for calculation were as follows: test family - t-test, statistical test - the difference between two independent means (two groups), a priori power analysis - tails: two, effect size: 0.8, α error probability: 0.05, power (1-β error probability): 0.85. The calculated sample size was 60 (30 participants in each group). To account for potential dropouts and reduce sampling error, the sample size was increased by 10%.Detailed interviews were conducted to screen potential participants. Individuals with a history of drug use, musculoskeletal ailments, cardiovascular diseases, or neurological problems that could impact normal gait patterns were excluded from the study.A convenience sample of 67 participants, comprising 30 non-smokers and 37 smokers, was recruited for the study. The sample included 41 males and 26 females. Participants' age ranged from 22 to 32 years, height ranged from 162 to 181 cm, weight ranged from 65 to 95 kg, and body mass index ranged from 22 to 25 kg/m². Non-smokers had never smoked, while current smokers had a smoking history of two or more years.A portable electronic self-calibrated stadiometer cum weighing scale  was used to the measured height, weight, and body mass index of each participant. Physilog 4 from GaitUp  was utilized to determine participants' gait parameters. The Physilog from GaitUp is a valid and reliable instrument for measuring spatiotemporal gait characteristics ,18.The high-quality 3D accelerometer, 3D gyroscope, and barometric pressure sensor (Physilog4 from GaitUp) were securely attached to the participants' upper feet using straps to ensure stability during measurements. It provided raw 3D acceleration data at a sample rate of 128 Hz. To measure synchronized Physilog data, the left and right leg sensors were turned on and connected to the network using the main button within a short time interval. Each sensor began recording data upon pressing the start button and synchronized with the master sensor. The green light on each sensor started blinking to indicate synchronization with the master sensor. The measurement was initiated by pressing the main button on the sensors and stopped by pressing the button until the LED turned orange.The gait recording procedure was standardized for each participant in the study. The examination technique began with an interview of each participant, followed by the completion of self-administered questionnaires at their own convenience. A 20-meter straight, flat, and hard surface was chosen for the walk. Physilog inertial sensors were attached to the foot of each participant. Participants were instructed to walk at their own pace on the straight path. The Physilog application for gait analysis was installed on a tablet used by the researcher, and a Bluetooth connection was established with the sensors. The previously described experimental setup was utilized to record three trials for each participant. The average of three trials was calculated and used for further analysis. The variables of interest in this study included general parameters of gait  and temporal parameters of gait .The data analysis was performed using the Statistical Package for the Social Sciences (SPSS), version 26.0, developed by SPSS Inc., located in Chicago, IL, United States. Data were carefully examined before the analysis to identify outliers and missing values. The normality of the data distribution was assessed using the Shapiro-Wilk test, with a significance level at p>0.05. The anthropometric characteristics and gait parameters were analyzed using the independent t-test (two-tails). A p-value of ≤0.05 was considered statistically significant.When comparing genders separately, there were significant differences in cadence (p=0.001), stride length (p=0.001), stride velocity (p=0.001), swing (p=0.001), flat foot (p=0.001), pushing (p=0.030), and double support (p=0.001) between non-smoker males and smoker males . HoweverIn the female subgroup, significant differences were observed in cadence (p=0.001), stride length (p=0.001), stride velocity (p=0.001), stance (p=0.005), foot flat (p=0.001), pushing (p=0.008), and double support (p=0.001) . In contWhen comparing genders, significant differences were observed across all gait parameters , except for the swing phase . Male paThe study aimed to determine differences in gait parameters between non-smokers and smokers. The results revealed significant differences between these two groups in gait parameters. Non-smokers had a higher cadence, longer stride length, longer stance, larger swing phase, and larger pushing than smokers. Smokers had higher stride velocity, higher loading phase, greater foot flat, and bigger double support than non-smokers.Furthermore, when comparing genders, significant differences were observed in all gait parameters  except for the swing phase. Male participants had greater mean values of cadence, stride length, stance, swing, and pushing than females. Female participants had greater mean values of loading, foot flat, and double support than male participants.et al. proposed that race may influence gender differences in gait characteristics, even though there were no significant differences in spatiotemporal parameters between genders [et al. revealed no differences in spatiotemporal parameters between females and males. Step and stride lengths were statistically higher in females than in males [These findings are consistent with previous studies investigating gender-specific differences in gait parameters. Their findings are aligned with our findings as young, healthy females have shorter stride lengths and slower gait speed while walking at their own pace than healthy young males, largely due to their shorter height . A study genders . Smith ein males . We beliet al. [Our study findings are consistent with the research conducted by Verlinder et al. , who invet al. –25. Bettet al. .One investigation found that greater pack-years of smoking were associated with poorer gait phases and pace, resulting in slower and smaller steps with longer double support . SimilarA study found significant gait speed impairment among women associated with smoking status . CurrentFollowing a physical exercise intervention for smokers and non-smokers, improvements in the walking economy were observed, with a 9% increase in smokers and an 11% increase in non-smokers. Significant differences were found between smokers and non-smokers for the total walking distance during the six-minute walk test, pain-free walking distance, and daily physical activities . AdditioSmoking impacts physical performance by affecting vascular function and metabolism . In addiThis study had some limitations. First, its cross-sectional design precludes establishing a causal relationship between anticipated factors and functional performance outcomes. Second, the sample size was smaller than in other epidemiological and prevalence studies, which may account for the statistically insignificant results. The authors did not examine smoking history, which may have affected the outcome variable favorably. Despite these limitations, the study has notable strengths. The gait characteristics were measured using valid and reliable tools, ensuring the accuracy of the data collected. In clinical settings, these gait parameters can be considered vital signs.Our results showed significant differences in the overall and temporal gait parameters between non-smokers and smokers. Non-smokers had a higher cadence, longer stride length, longer stance, larger swing phase, and larger pushing than smokers. Smokers had higher stride velocity, higher loading phase, greater foot flat, and bigger double support than non-smokers. Functional performance evaluations and quantitative data on gait disorders benefit greatly from gait characteristics. Longitudinal studies with high sample numbers are needed in future research to investigate the impact of various confounders on gait parameters.


Mycobacterium tuberculosis (M. tb) infection has focused on T cells and macrophages, as their role in granuloma formation has been robustly characterized. In contrast, the role of B cells in the pathophysiology of M. tb infection has been relatively overlooked. While T cells are well-known as an essential for granuloma formation and maintenance, B cells play a less understood role in the host response. Over the past decade, scarce research on the topic has attempted to elucidate the varying roles of B cells during mycobacterial infection, which appears to be primarily time dependent. From acute to chronic infection, the role of B cells changes with time as evidenced by cytokine release, immunological regulation, and histological morphology of tuberculous granulomas. The goal of this review is to carefully analyze the role of humoral immunity in M. tb infection to find the discriminatory nature of humoral immunity in tuberculosis (TB). We argue that there is a need for more research on the B-cell response against TB, as a better understanding of the role of B cells in defense against TB could lead to effective vaccines and therapies. By focusing on the B-cell response, we can develop new strategies to enhance immunity against TB and reduce the burden of disease.Historically, research on the immunologic response to  M. tb [M. tb primarily affects the lungs but can cause disease in any part of the body [M. tb can lead to the bacteria being isolated within a granuloma, also known as latent TB [M. tb but experiencing no clinical symptoms of active TB (ATB) [TB is caused by an airborne pathogen of the organism M. tb . M. tb pthe body . The lunthe body . Infectiatent TB . Latent TB (ATB) . Only ATTB (ATB) . In 2018TB (ATB) . It is eTB (ATB) . RecurreTB (ATB) ,6.M. tb is an intracellular pathogen, meaning that it can survive and replicate inside host cells. Therefore, humoral immunity provided by B-cell-mediated Abs production is thought to be inefficient in accessing and harming intracellular bacteria [Despite the significant burden of disease, the only effective TB vaccine available is bacillus Calmette-Guérin (BCG) . The BCGbacteria . HoweverM. tb invades, the innate immune cells recognize M. tb by pathogen-associated molecular patterns (PAMP) via an immune cell pattern recognition receptor (PRR). Within minutes of M. tb exposure, the innate immune cells create an inflammatory response [M. tb, are especially pathogenic due to their ability to evade host immune defenses. As an intracellular pathogen, M. tb must gain access to dividing immune cells, such as macrophages and DC, to divide [Anatomical and physiological barriers provide the first line of defense against invading pathogens. The subsequent line of protection is the innate immune response, which includes macrophages, neutrophils, mast cells, natural killer cells, dendritic cells (DC), and eosinophils ,11. Onceresponse . Bacterio divide .M. tb, the pathogen reaches the alveoli , which leads to the loss of NAD+ in the M. tb-containing macrophages, which inhibits necrotic cell death of the macrophage  are the sites for Abs diversification, maturation, and antigen-dependent clonal expansion. Thus, they are essential to humoral immunity . GC are trocytes . The LZ-trocytes . Early ttrocytes . Howevertrocytes . A studytrocytes . A follotrocytes . OverallWhile GC have been characterized extensively in secondary lymphoid tissue, their presence in TB granulomas is a relatively new finding . GC are M. tb infects the host airway immune cells, evades phagocytosis, and persists and replicates intracellularly leading to host cell necrosis and intercellular dissemination of the pathogen. To prevent further dissemination of M. tb, the host immune system responds with granuloma formation, the hallmark of M. tb infection. Granulomas are defined as aggregates of immune cells that produce various cytokines to limit bacterial growth. Granuloma formation begins when infected macrophages produce cytokines leading to the activation of CD4+ T lymphocytes, which then produce proinflammatory cytokines, such as IFNγ and TNFα [M. tb and granuloma formation.During the acute phase of TB, and TNFα . These cand TNFα . AlthougM. tb and demonstrated that lung granulomas of infected NHPs contained significantly more activated B cells compared to noninfected NHPs [Phuah et al. (2012) used several biomarkers to characterize the composition of the granulomas found in nonhuman primates (NHP) infected with ted NHPs . A studyted NHPs . ClassicSeveral studies suggest the granuloma structure formed during TB infection closely resembles that of GC found in secondary lymphoid tissues. Various studies have highlighted structural and functional similarities between the GC in secondary lymph nodes and granulomas in TB infection. Structurally, granulomas contain a necrotic core surrounded by a layer of immune cells that form a physical barrier to contain the infection. Phuah et al. used several biomarkers to study the structure of granulomas in NHP and found CD20+ B cells clustered toward the periphery of granulomas similar to that of GC . The B cM. tb-infected mice served various complex functions in eliciting an immune response against TB. B cells within granulomas were found to produce the chemokine CXCL13, which recruits CXCR5+ T cells to the infection site, and promotes formation of follicle-like structures within granulomas [M. tb and found to have an increased bacterial burden, enhanced inflammation of lung tissue, and a decreased number of CXCR5+ T cells within granulomas. The B cells found in M. tb-infected lungs were also shown to upregulate the expression of Sftpc and Ccr7. Sftpc is an important gene that codes for a surfactant protein usually found in lung epithelial cells. B-cell expression of Sftpc confirms their localization to lung parenchyma during an ATB infection. Ccr7 is a vital chemokine that promotes B-cell follicular development within TB granulomas [M. tb [Given the structural similarities between B cells found in GC and TB granulomas, further studies were conducted to better understand the role of B cells within granulomas . Choreñoanulomas . To confanulomas . Similars [M. tb .M. tb infection [M. tb, has been thought to exclusively elicit a cellular immune response, B cells and the humoral immune response help complement the cellular immune response to contain and overcome intracellular infections.The acute phase of TB is primarily understood to be a T-cell- and macrophage-driven process . Howevernfection . AlthougM. tb-directed Abs can regulate effector functions, such as neutralizing antigens, opsonization, and ADCC [M. tb is a non-toxin-producing, intracellular pathogen, neutralizing Abs are thought to bind to epitopes on M. tb and play an essential role in the Abs-mediated clearance of M. tb [M. tb led to increased phagocytosis by macrophages and enhanced complement binding of C3 and C4 [M. tb is primarily an intracellular pathogen, M. tb re-enters the extracellular space when infecting other host cells. During this phase, IgG ADCC limits growth of M. tb and plays an important role in host defense [M. tb through various mechanisms while supporting the cellular immune response [In addition to acting as antigen presenting cells, B cells function to produce Abs directed against pathogens. Once activated, and ADCC . Althougof M. tb . Abs proof M. tb . A recen3 and C4 . Althoug defense . Purifie defense . Abs havresponse .M. tb growth and inflammation of the lungs [M. tb-specific Abs within granulomas. Furthermore, infected tissues contained higher levels of TB-specific IgG. These findings show that B cells are not only present but actively generating M. tb-specific Abs at the site of infection [M. tb antigens have no role in clinical therapy or prophylaxis and a minor role in TB vaccines [In the clinical setting, passive administration of human IgG has been shown to control he lungs ,65. Phuanfection ,47. In anfection . Researcnfection . Despitenfection . In addinfection . Currentvaccines . As suchM. tb mice. To investigate the impact of IL-6 produced by B cells, a mouse strain with specific IL-6 deficiency in B cells was established on the B6 genetic background [flox/flox mice [cre/cre) [Studies have shown that IL-6 inhibition leads to an increased risk of TB; it is often considered one of the essential cytokines during infection ,69. IL-6ckground . Mice wilox mice  with CD1cre/cre) . The stucre/cre) . This sucre/cre) . Finallycre/cre) . Howevercre/cre) .M. tb. Then, they investigated B-cell depletion and its impact on granuloma T-cell cytokine secretions, such as IL-17, TNF-α, IFN-γ, IL-10, and other immunoregulatory cytokines [M. tb-infected macaques. By contrast, B cells significantly affected cytokines and inflammation levels [M. tb infection. However, the study is limited regarding inclusion criteria as all patients were in the early phase of the M. Tb disease. Finally, a study by Cantini et al. aimed to evaluate TB reactivation in patients who have rheumatoid arthritis receiving rituximab and other non-anti-tumor necrosis factor agents [Rituximab is used clinically to treat systemic lupus erythematosus and multiple sclerosis ,52. Studytokines . Resultsytokines . Howeverytokines . The stun levels . Lastly,n levels . This dar agents . Resultsr agents . These cr agents .The role B cells play in the acute phase response to a TB infection has been corroborated by multiple sources; however, there is conflicting research on their role in the later phase of the disease. Whether their role in later infection is limited or even detrimental has yet to be solidified by research. An experiment by Suraj Parihar showed that IL-4-responsive B cells increased mice’s mycobacterial burden and lungs . Most otM. tb burden but enhanced cachexia and shortened the animals’ life span compared to controls  explored the role of B cells during late TB infection, after 16 weeks. Results showed that TB susceptible mice had a significant decrease in B cells and B-cell follicular formation at week 16 post infection, which led to an increase in inflammatory cytokines genes encoding IL-1, IL-11, IL-17, and TNF-α when compared to TB-resistant mice . Moreovecontrols C  vaccine has been administered in other parts of the world to develop protection against occurred .M. tb would be the production of anti-TB vaccines [M. tb. Studies show that the passive transfer of monoclonal Abs against M. tb antigens can aim to enhance a host’s immune response should it be exposed to the pathogen [M. tb: (1) Passive transfer of monoclonal Abs against the M. tb antigen alters the disease course such that there is an overall benefit to the host; (2) the existence of those specific Abs leads to decreased susceptibility to the particular disease (TB); and (3) there is increased susceptibility to the particular disease (TB) in those who are deficient in the Abs against M. tb. [While a targeted antibiotic regimen has been developed and used in many of those afflicted with TB, clinicians and researchers still maintain that a more cost-efficient and effective way to combat vaccines . This bepathogen . In addit M. tb. .M. tb VAC HK, in mice and NHP. They found that the vaccine was able to recruit mucosal Abs and aid with M. tb opsonization and improve protection against TB [M. tb Ag85b or B. anthracis protective antigen [M. tb-specific AM. Mice immunized with the vaccines were found to have a lower bacterial burden and a longer life span. Dijkman et al. (2019) further studied the role of the humoral immune response against M. tb specifically in the pulmonary mucosa of NHP [M. tb [M. tb disease process. The concluded interpretations of these independent studies therefore point to the postulation that anti-TB vaccines that induce Abs-mediated immunity can result in enhanced protection against the life-threatening disease.The goal of a vaccine is to induce long-term immunological memory . Despiteainst TB . Additio antigen . Their ra of NHP . PulmonaP [M. tb . With thM. tb infection. Despite the potential role of B cells in combating M. tb, studies on B cells and M. tb infection are limited. In contrast to the previous understanding of B cells and the humoral immune response against intracellular pathogens, we show that emerging research supports that B cells may attenuate the life span of many intracellular organisms, such as chlamydia trachomatis and salmonella enterica. Regarding TB, recent data have demonstrated divergent humoral signatures, unique Ab Fc function, and distinct Abs glycosylation patterns when comparing individuals with ATB and LTB infection. Furthermore, it has been shown that animals infected with TB produce M. tb-specific Abs in the lungs.This review aimed to determine the significance of the humoral immune response against Moreover, new evidence suggests that the humoral immune response may partially drive granuloma formation, formerly thought to be exclusively a macrophages and T-cells-mediated process. Experiments in NHP demonstrated that granulomas with fewer B cells had worse outcomes and a higher disease burden. Furthermore, studies reveal striking similarities when comparing the structure of granulomas and GC, such as B cells found in the periphery of granulomas. However, unlike GC, these B cells were not shown to be actively dividing. In addition, B cells within granulomas produced CXCL13, which recruits CXCR5+ T cells to create follicle-like structures within the granuloma. We can conclude that granuloma formation and maintenance may require B cells. However, additional analysis and testing are needed.The number of experimental models showing the importance of T cells and macrophages is exhaustive. Our review shows that B cells are actively involved in this phase of the disease by creating IgG-dominated opsonization leading to increased phagocytosis. In cytokine production experiments, B-IL-6KO strains had a significant decrease in total IL-6 production, leading to increased TB susceptibility. Finally, B-cell depletion in the chronic stage of infection resulted in neutrophil inflammation and contributed to significant lung fibrosis. Overall, this suggests that B cells help fight TB in the acute phase of infection by creating proinflammatory cytokines and Abs. However, in the later stage of the disease, B cells have an anti-inflammatory effect by decreasing lung fibrosis and neutrophil inflammation.M. tb challenge. By contrast, more recent studies show that inactivated M. tb vaccines in mice and NHP were able to recruit antibodies that aid in the protection against TB. These conflicting results question the validity of mice and other animal studies of the B-cell response in a TB infection due to the divergent structure of immune organs compared to humans. Despite these controversial findings, a growing body of the literature shows the importance of B cells in combating a M. tb infection. Further research is needed to address these issues.Despite these findings, several studies resulted in somewhat controversial conclusions, such as the rituximab study, which showed that TB risk was negligible for patients taking rituximab. Moreover, vaccine studies are even more conflicting, with some studies concluding that mouse monoclonal Abs showed no protection against a M. tb and protect the host. However, more research is vital to better understand the contribution of B cells to the immune response against TB. Therefore, the role of B cells in TB immunity may lead to the development of novel therapies to help combat the global TB threat.The role of B cells in TB is poorly understood, and research is often contradictory. Current research seems to suggest a phase-dependent role of B cells in fighting a TB infection. Most research is in agreement that B cells are critical for the initial phase of infection, including cytokine production that activates neutrophils and macrophages and the subsequent granuloma formation. Research is less detailed about how B cells behave after the first 13 weeks of infection. In conclusion, developing research suggests that B cells play a critical role in killing intracellular organisms by creating cytokines and Abs to help eliminate