Antimicrobial resistance is a phenomenon that the present-day world is witnessing that poses a serious threat to global health. The decline in the development of novel therapeutics over the last couple of decades has exacerbated the situation further. In this scenario, the pursuit of new alternative therapeutics to commonly used antibiotics has gained predominance amongst researchers across the world. Antimicrobial peptides (AMPs) from natural sources have drawn significant interest in the recent years as promising pharmacological substitutes over the conventional antibiotics. The most notable advantage of AMPs is that microorganisms cannot develop resistance to them. Insects represent one of the potential sources of AMPs, which are synthesized as part of an innate immune defence against invading pathogens. AMPs from different insects have been extensively studied, and silkworm is one of them. Diverse classes of AMPs  were identified from silkworm that exhibit antimicrobial property against bacteria, fungi and viruses, indicating their potential therapeutic benefits. This review briefs about the immune responses of silkworm to invading pathogens, the isolation of AMPs from silkworms, AMPs reported in silkworms and their activity against various microorganisms. By the year 2050, infections resistant to antibiotics are anticipated to result in the deaths of up to ten million people annually and are likely to cost the world economy over $100 trillion. Considering the gravity of the situation, the World Health Organization listed antimicrobial resistance as one of the top ten health threats being faced by humanity worldwide [Antimicrobial resistance is considered to be one of the major threats to global public health. This menace has emerged in the recent decades as a result of antibiotic misuse in human and animal healthcare, particularly in most of the developing nations ,2,3. Antorldwide ,5.Furthermore, adding to the difficulty of this endeavour, the development of new classes of antimicrobial agents has declined over the past thirty-five years. This might be attributed to the long-standing conventional approaches used in the search for antibiotics or perhaps the fact that many of the natural structures that exhibit antimicrobial action have already been identified ,7,8. As Bacillus by Rene Dubos in 1939, a number of AMPs have been isolated and characterized [https://aps.unmc.edu/home: accessed on 27 February 2023). According to a research report dated in 2020, seven AMPs received approval from the U.S. Food and Drug Administration for usage and are commercially available [In recent years, there has been a tremendous increase in the screening of natural products for the development of novel therapeutics. Natural compounds exhibit vast chemical diversity, making them important and reliable sources of novel medications . AMPs arcterized . Approxicterized   were also reported to be susceptible to AMPs from insects [Insect-derived AMPs are reported to inhibit the growth of bacterial pathogens belonging to various genera. Prominent among them are multiple drug resistant (MDR) bacteria including  insects .AMPs are the most sought after therapeutic compounds due to their characteristics, which include low toxicity to humans and animals; high specificity and improved efficacy against target microbes when compared to conventional antibiotics; and, most importantly, the fact that majority of microbes cannot develop resistance to AMPs. Most of the AMPs disrupt the bacterial cells via nonspecific interactions with their membrane surface. AMPs attack numerous low affinity targets such as bacterial membranes, as opposed to traditional antibiotics, which act through a specific high-affinity antimicrobial target and can result in microbial resistance. The swift microbicidal property of AMPs is also considered as a factor that prevents evolution of resistance. These might be the most probable reasons for minimal/restricted emergence of bacterial resistance against most of the AMPs ,25. HoweAMPs, apart from being used in medicine and allied fields, also find their application in various sectors viz., food processing, animal husbandry, aquaculture and agriculture . BesidesBombyx mori has long been utilized to produce silk and is economically significant in many countries that practice sericulture. Silkworms are susceptible to infection by a variety of microbial pathogens including bacteria, fungi and viruses. Apart from microbial pathogens, silkworms are also vulnerable to pests such as Exorista bombycis that infects the larval stage and Dermestid aeter that attacks pupal or moth stages. The causal microorganisms as well as specific symptoms of different silkworm diseases and the images of infected silkworms are depicted in The silkworm B. mori has an efficient innate immune system to fight against various pathogens and pests. The first line of defence against pathogenic infection in silkworms is provided through physical barriers such as integument (a protective exoskeleton) and the peritrophic matrix, a semipermeable membrane layer that protects the midgut epithelium [BmPGRP2-1 (a transmembrane protein) and BmPGRP2-2 (an intracellular protein) are two important PGRPs in B. mori that activate the immunodeficiency (Imd) pathway and suppress the PTEN-phosphoinositide 3-kinase (PI3K)/Akt signalling, respectively, in response to B. mori nucleopolyhedrovirus (BmNPV) infections [Like any other insect, silkworms lack an adaptive immune mechanism and solely depend on their innate immune system to combat the invading pathogenic microorganisms/pests . The silithelium ,39. Uponithelium ,41,42. Tithelium ,44. The ithelium . BmPGRP2fections .Cellular immunity is mainly carried out by haemocytes (free circulating immunosurveillance cells), which are classified on the basis of morphology and size into five types: prohemocytes (Pr), granulocytes (Gr), plasmatocytes (Pl), spehrulocytes (Sp) and oenocytes (Oe) ,45,46,47B. mori upon bacterial, viral, fungal or parasitic infection. The melanisation reaction is initiated on the recognition of pathogens by host PRRs, which activates the serine protease cascade. The pro-PPAE (pro-prophenoloxidase activating enzyme) is converted to active PPAE via serine proteases. PPAE catalyse the conversion of PPO to active phenoloxidase (PO) enzyme. Further, PO activates the catalytic reaction of phenol-oxidation to quinones leading to the formation of insoluble melanin [B. mori. It is reported that specific serpins are induced significantly in silkworms challenged with bacterial pathogens. One such protein, serpin-6 (BmSP6) regulated the immune pathway in silkworms by inhibiting the activation of PPO and induction of an AMP, peverin-2 [Melanisation is an independent pathway induced in haemolymph, haemocytes and other immunocompetent tissues of  melanin . Serine everin-2 . Accordieverin-2 ,62.The humoral immune response is mainly activated in the fatbody and, in some instances, in other immunocompetent tissues such as integumental epithelium, midgut epithelium and haemocytes ,65,66,67B. mori, has five instars during which the larvae go through four moults. Silkworms are challenged with pathogens to isolate AMPs during their fifth instar as the duration is longer (6–8 days), which allows for enough time to develop infection. Furthermore, silkworm’s fat body content is at its peak during this instar, which is the primary source of AMPs [The larval stage of silkworm,  of AMPs ,76. Upon of AMPs . The pro of AMPs , Thermo  of AMPs , PEAKS [ of AMPs , Maxquan of AMPs ,82. The  of AMPs .AMPs from silkworm are low molecular weight proteins  among which a majority of them exhibit broad spectrum activity against different microorganisms. The different families of AMPs characterized and reported in silkworms, their characteristics, structure, mode of action and efficacy against bacterial species are presented in Hyalophora cecropia moth infected with bacteria [B. mori, five types of cecropins are found including cecropin A, cecropin B, cecropin C, cecropin D and cecropin E. A total of eleven Bmcec genes  encoding cecropins are reported in silkworms [B. mori [H. cecropia cecropins at lower concentrations interact with membranes to form channels or pores, affecting cellular electrolyte balance, thereby causing cell death [Cecropin, α-helical linear AMP (37 amino acids) lacking cysteine residues was first isolated from bacteria . In B. milkworms . Another[B. mori ,86. Cecr[B. mori ,88. Alth[B. mori . Howeverll death .B. mori cecropins, CecXJ-37C and CecXJ-37N with an amino acid addition on C-terminal, are also reported to be active against diverse bacterial strains [Aspergillus spp., Fusarium spp. and yeasts indicating antifungal properties of this AMP [B. mori cecropins did not exhibit any cytotoxic or haemolytic effects at concentrations up to 200 µM, but they inhibited growth of microbial pathogens at much lower concentrations [Cecropins at very low concentrations exhibit antibacterial activity against a wide range of Gram positive and Gram negative bacteria . Two mod strains . Cecropithis AMP . These pthis AMP . Reportstrations .B. mori inhibited growth of hepatocellular carcinoma (HCC) cell line Huh-7 cells and induced apoptosis in HCC cells [Apart from antimicrobial properties, cecropins are reported to selectively induce apoptosis in cancer cells. CecropinXJ, a newly isolated cationic AMP from CC cells . CecropiCC cells . CecropiCC cells .Sarcophaga peregrina, the flesh fly [BmDefensinA found in B. mori genome is a defensin ortholog of Spodoptericin. A group of researchers reported expression of BmDefensinB gene in B. mori after infection with E. coli, Bacillus subtilis and Beauveria bassiana [B. subtilis, B. thuringiensis, B. megaterium Micrococcus luteus, S. aureus and Aerococcus viridians [B. mori is also reported, which is identical to BmDefensinA and is close to galiomicin and spodoptericin [Defensins are cysteine containing peptides that were first reported from lesh fly . Defensilesh fly . BmDefenbassiana . Defensiiridians . A defenptericin .Defensins inhibit bacterial growth by membrane disruption and through the formation of voltage dependent anion-selective channels in cell membranes ,102,103.B. mori haemolymph. It was found to be active against Gram positive bacteria S. aureus [B. mori, a total of twelve genes encoding moricin have been reported and divided into two subfamilies on the basis of sequence similarity. Out of twelve moricin genes, four belong to subfamily BmmorA (A1 to A4) and eight belong to subfamily BmmorB (B1 to B8) [Moricin, a cationic, amphipathic α-helix AMP shows the presence of charged amino acid residues after every three to four amino acids, which is responsible for its antimicrobial properties against bacteria and few strains of yeasts. Moricin consists of 42 amino acid residues and was first isolated from the . aureus . In B. m1 to B8) .B. mori. Moricin contains N-terminal fragment (5–22 amino acids), which is amphipathic, α-helical and is the active site for antibacterial activity. The C-terminal region of moricin initially interacts with the surface of bacterial membrane and then permeability of the membrane is altered by N-terminal amphipathic a-helix. It is reported that the voltage-dependent pores could be formed through interaction between three or more amphipathic α-helices spanning a lipid membrane [A very limited amount of literature is available on the mechanism of pore formation in bacterial cell membrane by moricins from membrane ,105. MorH. gloveri pupae [E. coli strains possessing smooth LPS. Reports suggest that the binding of gloverins to the inner part and Lipid A region of LPS is required for its activity. A conformational change occurs in the gloverins when they penetrate the hydrophilic regions of LPS layer and interacts with negatively charged hydrophobic regions made of lipid A [Gloverins are glycine-rich AMPs of molecular weight 8–30 kDa and were first reported from haemolymph of giant silk moth ri pupae . Gloveri lipid A ,107. BmGB. mori, four genes encoding gloverins  were identified. All four gloverin genes were activated by E. coli, B. subtilis, and Salmonella abortusequi while the expression of gloverin genes was reduced when challenged with S. aureus [B. bassiana [E. coli JM109 and Pseudomonas putida) by enhancing the cell membrane permeability [In silkworm . aureus . The dif. aureus . BmGlovebassiana . It is aH. cecropia pupae inoculated with bacteria [E. coli and Trypanosoma brucei [E. coli and Citrobacter freundii) and also antifungal activity  [Attacins are low molecular weight (20–23 kDa) AMPs that were first isolated from the haemolymph of bacteria . On the a brucei , whereaslbicans) . Attacinlbicans) ,114.B. mori haemolymph challenged with E. coli. Lebocin family consists of four protein encoding genes . The expression of lebocin genes is induced by LPS in haemocytes and fatbody [Acinetobacter sp. and E. coli), Gram positive bacteria and fungi. Lebocin-B and Lebocin-C isolated from another lepidopteran insect, Manduca sexta, differ from B. mori Lebocin and are reported to possess antibacterial activity against Serratia marcescens, S. typhimurium (Gram negative); S. aureus, B. cereus (Gram positive) and Cryptococcus neoformans (fungi) [Lebocins (32 amino acids) are proline-rich AMPs with O-glycosylated residues that were isolated from  fatbody . Lebocin (fungi) .B. mori, AMPs have also been identified from the nonmulberry silkworms belonging to the family Saturniidae, namely Antheraea assamensis (muga), Antheraea mylitta , Antheraea pernyi (temperate oak tasar), Antheraea yamamai (Japanese oak tasar) and Samia cynthia ricini (eri).In addition to the AMPs from the domesticated mulberry silkworm, S. cynthia ricini. A cDNA clone of Scr-gallerimycin (AB366558) gene encodes 74 amino acids and the gallerimycin protein has 6.21 kDa of calculated molecular mass and 7.6 pKa [B. mori and Trichoplusia ni lebocin precursor proteins [S. cynthia ricini challenged with bacteria. Both the attacin genes were coded for 233 amino acids and shared 98% identity at protein level, whereas at nucleotide level, 92% identity was reported [A. pernyi whose expression level significantly increased in fatbody upon E. coli infection [C. albicans [A. mylitta, a glycine-rich antimicrobial peptide (GGGGGGHLVA) was reported to be active against MDR E. coli associated with urinary tract infections [2-Gln-Ala-Lys-COOH (QAK) was reported to be isolated and purified from haemolymph collected from immunized A. mylitta. Acetylated and non-acetylated QAK peptide exhibited antibacterial activity against E. coli and S. aureus [A. yamamai exhibited antimicrobial activity against Gram negative bacteria , Gram positive bacteria  and fungi , indicating its broad spectrum potential. The authors reported that MIC values against the tested Gram negative bacteria, Gram positive bacteria and fungal strain ranged between 1–2 µg/mL, 64–128 µg/mL and 64 µg/mL, respectively [A. mylitta and fractionated by HPLC. The fractions were assessed for their antibacterial activity against MDR bacteria including E. coli, P. aeruginosa and B. pumilus. It was found that fraction II exhibited antibacterial activity against E. coli (zone of inhibtion-9 ± 0.35 mm) and P. aeruginosa (6.5 ± 0.40 mm), whereas fraction III was active against only B. pumilus (7.5 ± 0.30 mm) [An antifungal peptide named gallerimycin is reported to be isolated from the fatbody of  7.6 pKa . A lebocproteins . The cDNreported . Anothernfection . A glovealbicans . In A. mfections . A tri-p. aureus . A cecroectively . In anotThe AMPs isolated from natural sources are generally unstable, and it is therefore imperative to determine their stability before going ahead with application in various fields. AMPs are affected by several factors such as metal ions, temperature, pH and proteases. Metal ions affect the self-assembly and activity of AMPs, while pH may have varied effects depending upon the charge of the peptides . The majIn order to overcome the influence of different factors mentioned above, the identified bioactive peptides could be synthesized chemically through solid-phase peptide and peptide synthesis in solution. Chemical synthesis of AMPs is advantageous over extraction of AMP from natural sources, as synthetic peptides are easy to modify as per the specific requirement ,133. Morhttp://dramp.cpu-bioinfor.org/; accessed on 27 April 2023) indicate 22,480 submissions, including natural and synthetic AMPs (6105 No’s), patent AMPs  and 77 AMPs in preclinical or clinical stages of drug development. Forty three percent of the 77 AMPs are in the preclinical stage and 33 peptides are in clinical trials. Six peptides failed during phase III studies, while one was denied permission [Streptomyces roseosporus and is employed in the treatment of complicated skin and skin structure infections. Daptomycin exhibited vast antibacterial activity against methicillin-susceptible and methicillin-resistant S. aureus, Vancomycin susceptible Enterococcus faecalis and different Streptococcus species [S. aureus, different strains belonging to the genus Streptococcus and E. faecalis [The most recent AMP data collected from the DRAMP  database (rmission . Althougrmission . In the  species . Oritavafaecalis .The antibiotic resistance crisis has led to exploration for novel therapeutics globally. AMPs have the potential to be an effective treatment method against drug-resistant bacteria. A variety of animals, including insects, that manufacture AMPs as a component of their innate immune system are being studied for novel AMPs, and silkworm is one of them. The silkworm has one of the most extensive repertoires of structurally and functionally distinct AMPs with antimicrobial activity against different microorganisms. This study draws attention to silkworm as a possible source of various antimicrobial peptides and opens the door to new avenues for intervention and the development of naturally occurring bioactive compounds to address antibiotic resistance.Like many of the insect AMPs, silkworm AMPs also have limitations in terms of low bioavailability, possible haemolysis, susceptibility to proteases, cytotoxicity, high production costs and low expression which continues to restrict their usage in clinical applications. Even though it is essential to continue exploring for novel silkworm AMPs, more research is also required to overcome the limitations preventing their clinical applications. Emphasis should also be placed on the development of transgenic or genome edited silkworms for over expression of AMPs. However, before using the AMPs for different applications, safety must be ensured. Strategies may also be employed to design better synthetic AMPs based on the sequences of AMPs from natural sources using in silico approaches.


Pyrus ssp.) belongs to Rosaceae and is an important fruit tree widely cultivated around the world. Currently, challenges to cope with the burgeoning sets of multiomics data are rapidly increasing. Here, we constructed the Pear Multiomics Database (PearMODB) by integrating genome, transcriptome, epigenome and population variation data, and aimed to provide a portal for accessing and analyzing pear multiomics data. A variety of online tools were built including gene search, BLAST, JBrowse, expression heatmap, synteny analysis and primer design. The information of DNA methylation sites and single-nucleotide polymorphisms can be retrieved through the custom JBrowse, providing an opportunity to explore the genetic polymorphisms linked to phenotype variation. Moreover, different gene families involving transcription factors, transcription regulators and disease resistance (nucleotide-binding site leucine-rich repeat) were identified and compiled for quick search. In particular, biosynthetic gene clusters (BGCs) were identified in pear genomes, and specialized webpages were set up to show detailed information of BGCs, laying a foundation for studying metabolic diversity among different pear varieties. Overall, PearMODB provides an important platform for pear genomics, genetics and breeding studies.Pear (Database URLhttp://pearomics.njau.edu.cn Pyrus, 2n = 2x = 34), belonging to the Rosaceae family and the Maloideae subfamily  have been domesticated to comprise the majority of modern cultivated varieties. As of 2021, the pear-cultivated area around the world has reached 1 399 484 ha, and the yield is ∼2 568 713.07 tons (http://www.fao.org/faostat/en/#home).Pear  was published with 512 Mb assembly size and 42 812 annotated genes, and genome evolution and important genes related to pear fruit quality  were resolved (http://peargenome.njau.edu.cn), but limited data and web services were provided. In the year 2014, the first genome representing European pear (P. communis cv. ‘Bartlett’) was published  identified, and 9.29 and 5.35Mb genome regions for Asian pears and European pears were found to be influenced by domestication, respectively . Analysihttps://www.ncbi.nlm.nih.gov/sra), including those from different pear tissues  of National Center for Biotechnology Information (NCBI) database  based on the integration of different types of omics data. A variety of online tools were developed for quickly accessing and analyzing pear multiomics data, including tools for gene search, sequence alignment, multifaceted genomic information browser, gene family search, transcriptional expression heatmap and synteny analysis. Particularly, biosynthetic gene clusters (BGCs) were identified at a genome-wide scale for seven pear varieties. The PearMODB will accelerate the integration of available pear multiomics data and facilitate future studies in pear genomics, genetics and breeding.Here, we constructed the Pear Multiomics Database (PearMODB) (http://peargenome.njau.edu.cn) (http://www.gigadb.org/) (https://www.rosaceae.org) (https://ngdc.cncb.ac.cn/gwh/) (https://figshare.com/) (https://www.ncbi.nlm.nih.gov/sra). Population variation data (SNPs) were obtained from a previous study (https://www.ncbi.nlm.nih.gov/bioproject/?term=PRJNA381668) (https://www.ncbi.nlm.nih.gov/bioproject/?term=PRJNA503323)  , 10. Gendb.org/) . ‘Nijisseae.org) , 9. ‘Cuicn/gwh/) , 8. ‘Zhore.com/) . RNA-seqA381668) . DNA metA503323) . The detTrimmomatic  was used−5. For collinearity analysis, first pairwise BLAST of whole-genome protein sequences between any two pear varieties and within each variety was performed using diamond software with an e-value cutoff of 1e–10 (Nelumbo nucifera) as an outgroup  in seven pear genomes using previously reported Arabidopsis NBS-encoding genes as query sequence (https://www.ncbi.nlm.nih.gov/cdd/) was used to identify the NBS structure domain. Finally, amino acid sequences of candidate NBS-encoding genes were submitted to NCBI-CDD to identify toll interleukin 1 receptor and leucine-rich repeat (LRR) domains. The COILS program was used to specifically detect the coiled-coil domains in candidate NBS-encoding genes at a threshold of 0.9  domain PF00931) sequence from the Pfam database. HMMER , which is a toolkit for constructing online genomic and genetic databases. The web pages of PearMODB are designed and established based on HTML+CSS+JavaScript and managed by Drupal (a website management system based on the PHP programming language). The data are loaded using a GMOD/Chado database schema . Flexible options were provided on this page to enable specific gene search for users  module was used to build an interface for the BLAST search  module was utilized to build a custom JBrowse for each of the seven pear varieties . ‘Expression Heatmap’ allows users to retrieve spatiotemporal expression profiles of any genes of interest  and allowed users to explore syntenic blocks within and between genomes of different pear varieties. The multiscale macro- and microsynteny visualization tools were available. Users could retrieve syntenic blocks for any gene ID of interest and browse gene pairs contained in the target syntenic block  was embedded into PearMODB (To provide web services for polymerase chain reaction (PCR) primer design, the primer3web interface  are available on the ‘Genome Data’ page. The protein sequences of TF, TR, PK and NBS-LRR family genes are available on ‘Family Genes Sequences’. We have also identified different modes of duplicated genes in genomes of seven pear varieties using DupGen_finder software , and thePyrus) multiomics data. PearMODB integrated different types of pear omics data and developed various online tools to cope with increasing requirements for diversified data analysis. Currently, PearMODB includes genome assembly, gene annotation, functional annotation, transcriptome, DNA methylation and population variation data. Available tools include gene search, BLAST, genome browser, expression heatmap, synteny browser and primer design. TF, TR, PK and NBS-LRR families are available for gene members search, providing a reference for gene function study and comparative genomics analysis. Different modes of duplicated gene pairs identified in seven pear genomes are also available on PearMODB. Specially, PearMODB provides an opportunity for investigating BGCs.PearMODB was developed to provide a comprehensive platform to access and analyze pear (https://www.rosaceae.org) (http://peargenome.njau.edu.cn) (http://www.peardb.org.cn/)  , Pear Ge.edu.cn)  and Pearorg.cn/) . The GDRorg.cn/)  and DNA org.cn/) . Third, org.cn/) . PearEXPorg.cn/) , which iThe future efforts would be made to exploit function modules involved in genome-wide association analysis between genetic polymorphisms and phenotypes, bridging gaps between genotype and phenotype. Large-scale population variation data such as SNP, copy-number variation and structural variation will be integrated to accelerate trait-related gene loci identification. Moreover, the construction of a gene co-expression network and a gene regulation network based on the integration of multiomics data will provide an important basis for the accurate prediction of gene function and the identification of molecular regulation pathways of important traits in pear.Driving by the rapid development of sequencing technology, large-scale omics data involved in pear pangenome, 3D-genome, proteome, metabolome, phenome and single-cell sequencing data will be available in the foreseeable future. PearMODB will be updated as more omics data become available and new tools for data analysis and visualization will be exploited. Collectively, PearMODB will be an important data hub for pear genomics, gene function and breeding studies.baad050_SuppClick here for additional data file.


Phyllostachys edulis. Overall, 81 TLP genes were identified in P. edulis; 166 TLPs from four plant species were divided into three groups and ten subclasses, with genetic covariance observed between these species. Subcellular localization in silico studies indicated that TLPs were primarily distributed in the extracellular. Analysis of the upstream sequences of TLPs demonstrated the presence of cis-acting elements related to disease defense, environmental stress, and hormonal responses. Multiple sequence alignment demonstrated that most TLPs possessed five conserved REDDD amino acid sequences with only a few amino acid residue differences. RNA-seq analysis of P. edulis responses to Aciculosporium take, the pathogenic fungus that causes witches’ broom disease, showed that P. edulis TLPs (PeTLPs) were expressed in different organs, with the highest expression in buds. PeTLPs responded to both abscisic acid and salicylic acid stress. These PeTLP expression patterns were consistent with their gene and protein structures. Collectively, our findings provide a basis for further comprehensive analyses of the genes related to witches’ broom in P. edulis.Thaumatin-like proteins (TLPs) are pathogenesis-related proteins with pivotal roles in plant defense mechanisms. In this study, various bioinformatics and RNA-seq methods were used to analyze the biotic and abiotic stress responses of the TLP family in  Phyllostachys edulis belongs to the Poaceae bamboo subfamily and is a crucial bamboo species with considerably high economic value [China has the highest distribution of bamboo and is rich in bamboo forest resources, which are vital assets for advancing the country’s forestry economy. ic value .Arabidopsis thaliana demonstrated that overexpression of BolTLP1 enhances tolerance to drought stress [Plants use complex mechanisms to protect themselves from pathogens. Thirteen classes of antifungal proteins have been identified based on their functional activities and structural characteristics . Pathoget stress . TLPs bet stress .Fusarium oxysporum [PnTLP2 gene isolated from Panax notoginseng is induced by Alternaria panax infection; the corresponding recombinant PnTLP2 protein has antifungal and defense effects [Populus tremula TLP (PtTLP) revealed that although PtTLP itself had no effect on pathogenic fungal strains, transgenic P. tremula with high PtTLP expression showed enhanced resistance to spot disease, and the leaf protein of the PtTLP overexpression model exhibited an apparent inhibitory effect on fungal growth in vitro [Medicago truncatula var. truncatula putative. All these proteins exhibited robust antifungal activities against Rhizoctonia solani, Alternaria alternata, Fusarium graminearum, Fusarium solani, Verticillium sp., and Phytophthora spp. [Studies on the biological function of TLPs in various plants have revealed their roles in defense against fungal pathogens. The ClTLP27 TLP was identified in watermelon that significantly inhibits the growth of various fungal pathogens, including xysporum . Alterna effects . A studyin vitro . Five TLora spp. . The mecora spp. .Aciculosporium take is highly prevalent in bamboo [A. take, apical bud growth is inhibited and early lateral bud germination into branchlets is stimulated, resulting in several lateral branches or axillary buds that form broom-like clusters that affect bamboo growth and even cause bamboo forest decline. Therefore, bamboo TLPs in the PR protein family that have antifungal activity must be systematically analyzed to effectively control witches’ broom in bamboo.Witches’ broom disease caused by n bamboo . In bambTLP genes have been identified and functionally analyzed in many plants, no such studies have been conducted in bamboo. As the whole genome of P. edulis was parsed, we used bioinformatics methods to identify TLP genes in P. edulis and analyzed gene structures, upstream cis-acting elements, chromosomal locations, conserved motifs, interspecific and intraspecific phylogenetic evolution, the three-dimensional (3D) structure of the encoded proteins, gene duplication, and transcriptional expression of these P. edulis TLPs (PeTLPs). This study aimed to comprehensively understand PeTLPs and explore their interactions with P. edulis and fungal invasion, thereby providing a basis for future research.Although TLP genes were identified from the P. edulis genome database. The expression levels of these genes are listed in PH02Gene08982) to 71,573.6 Da (PH02Gene13259). The isoelectric point values of these PeTLPs ranged from 3.89–8.68 and included 28 basic proteins and 53 acidic proteins. Subcellular prediction results showed that most PeTLPs were determined to be extracellular. PH02Gene35939, PH02Gene13257, and PH02Gene13259 were located in the plasma membrane, PH02Gene11792 in the nucleus, and PH02Gene11793 in the chloroplast.Eighty-one P. edulis showed high conservation among all 81 PeTLP sequences  contained no introns. Group 3 comprised the smallest PeTLP group, with 14 members. In group 3, only one gene (PH02Gene32525) possessed two untranslated regions (UTRs), whereas the remaining genes had no UTRs.To study the structural diversity of PeTLPs, their exon–intron structures and motif distributions were analyzed. A corresponding phylogenetic tree was constructed using previously identified PeTLP protein sequences, and its evolutionary relationships were evaluated. As shown in TLP gene sequences. The motifs differed between groups; however, members within the same group tended to exhibit similar motif patterns. For example, in group 1 (THN), motif 5 was identified as a characteristic motif. Motif 2 was a characteristic structure of group 2 (TLP). The group 3 (GH64-TLP-SF) superfamily possessed motif 7, except for PH02Gene50388. These results confirmed that the gene structure and motif patterns influenced phylogenetic diversity.The conserved regions of proteins form the basis of their function. To further reveal the structural diversity and functional characteristics of the PeTLPs, their motif patterns were investigated b. ThaumaP. edulis, A. thaliana, Oryza sativa, and P. tremula) and constructed phylogenetic trees  to more precisely demonstrate evolutionary and functional relationships of TLP genes between P. edulis and other species.To explore the evolutionary relationships of the TLP family among different species, we collected 166 TLPs from four plant species  and group 2 into three subclasses   .P. edulis, A. thaliana, O. sativa, and P. tremula TLPs. The largest TLP subclass was J, which contained 28 TLPs, and the smallest was I, which contained only seven members. In addition to subclass H, nine other subclasses comprised TLPs from both P. edulis and other plant species, indicating a close relationship between PeTLPs and TLPs from other species. Among the subclasses, there were homologous relationships between P. edulis, A. thaliana, O. sativa, and P. tremula in six subclasses . Overall, PeTLPs were primarily distributed in subclasses I, J, and F; however, subclasses I and J only exhibited homology between P. edulis and O. sativa, which were the two species that have been identified to have the highest homology.Multiple pairs of homologous genes were found among the PeTLPs were unevenly distributed on P. edulis scaffolds , one dicotyledonous herb , and one dicotyledonous woody plant (P. tremula), as shown in PeTLP genes exhibited syntenic relationships with those in O. sativa. The numbers of PeTLP genes with syntenic relationships with P. tremula and A. thaliana were 35 and 11, respectively. Scaffold 3 (PH03Gene31537), scaffold 6 (PH03Gene08982), scaffold 13 (PH02Gene11255), and scaffold 14 (PH02Gene32603) were homologous to all three species . Hormone-related elements were present in a large proportion of the PeTLPs (n = 79). Among the ABA-responsive elements, ABA response elements (ABRE) were present in many genes. Thirty-three genes contained three ABRE elements. PH02Gene05114, PH02Gene05115, PH02Gene08982, PH02Gene34215, PH02Gene40419, PH02Gene46317, PH02Gene48556, and PH02Gene50762 contained four hormone-related elements. Overall, 75 of these genes contained both hormonal and stress response elements. The findings indicate that PeTLPs may play an important role in forming broom-like branches and contribute to the corresponding resistance of P. edulis to witches’ broom.Cis-acting elements play crucial important roles in gene expression and reveal gene function. As shown in P. edulis in response to A. take infection were determined using ribonucleic acid sequencing (RNA-seq) technology. Overall, messenger ribonucleic acid expression in the apical buds, lateral buds, leaves, and stems of P. edulis was detected after infection with A. take. Healthy tissues were used as controls. A |Log2 fold-change (Log2FC)| > 1 and false discovery rate < 0.05 were the criteria to identify differentially expressed genes (DEGs).TLPs belong to the PR protein family, which primarily contribute to pathogen resistance. The response of bamboo to pathogens at the transcriptome level has not yet been reported. Thus, the transcript profiles of A. take predominantly infect bamboo buds, thereby inducing these buds to continuously differentiate to form witches’ brooms. However, this pathogen had minimal interaction with stems and leaves. The results of the present study are consistent with this phenomenon.As shown in PH02Gene34852, PH02Gene32892, PH02Gene35939, PH02Gene48556, PH02Gene28874, and PH02Gene28872 were more than four times higher in buds infected with A. take. PH02Gene48556 and PH02Gene48120 were notably upregulated in both buds and stems. Five genes  were notably downregulated (four times) in infected A. take. In particular, PH02Gene19402 was strongly downregulated in all four plant tissues. These findings indicated that these genes play crucial roles in the response of P. edulis to A. take infection.The relative expression levels of PeTLP expression levels in P. edulis buds after A. take infection, qRT-PCR was performed to verify the expression of 11 transcripts with notably different expression levels . Contrastingly, the expression levels of PH02Gene11255, PH02Gene28313, PH02Gene43141, PH02Gene19402, and PH02Gene31888 were notably lower in the diseased bamboo shoots. In particular, the expression level of PH02Gene31888 was 4.97× less than that in healthy shoots. Overall, the qRT-PCR results were consistent with the transcriptome data. The comparative statistical analysis results of the gene expression acquired by qRT-PCR and RNA-Seq in healthy and diseased buds were shown in To further investigate the differences in n levels . ComparePeTLPs expression profile of the transcriptome is shown as a heatmap , respectively. The leaf  heatmap .PeTLPs that mediated the corresponding stress response increased along with an increase in the function of these PeTLPs.Under NaCl treatment, ten and nine DEGs were upregulated, and five and fourteen DEGs were downregulated at 3 and 24 h, respectively. In the PEG treatment group, five and three DEGs were upregulated, and ten and nineteen DEGs were downregulated at 3 h and 24 h, respectively. As the duration of abiotic stress increased, the number of PeTLPs contained multiple hormone stress response sites. Therefore, we used the transcriptome data of different genes to verify whether their expression levels were induced by SA and ABA treatments , grapes (n = 33) [TLPs have been identified in various plants. These proteins play vital roles in the defense response of plants, as evidenced by the increasing number of studies. However, TLPs vary in quantity across different plants. In this study, 81 (n = 33) , and wes(n = 33) .PeTLPs to their corresponding scaffolds based on the location information of the P. edulis genome. The PeTLPs were unevenly distributed on the P. edulis scaffolds. Because of the relatively large size of the P. edulis genome, its assembly is not optimal. Many studies have analyzed the evolutionary relationships of TLP families among different species. The 118 TLPs from plants, animals, and fungi have been classified into nine major groups, with five groups containing most of the plant species [TLP genes from two dicotyledons and two monocotyledons into 10 subclasses  were found in subclass A, which contains members that reportedly participate in the ABA signaling pathway and abiotic stress responses [PeTLPs in subclass A, of which two genes were strongly differentially expressed following ABA treatment. Additionally, the functions of unknown PeTLP members can be predicted and verified based on the established functions of TLP gene family members from other species.Additionally, we mapped  species . Here, wbclasses . The disO. sativa and P. edulis had the most collinear genes, presumably because they both belonged to the Poaceae family. A previous study identified six tandem-replicated regions and 12 fragment-replicated gene pairs in melon [Previous studies on the phylogeny of TLPs have suggested that the family of plant TLPs is phylogenetically related and may have originated from the common ancestral genes of gymnosperms and monocots 130 million years ago . Collinein melon , which iCamellia sinensis TLPs are primarily localized in the cell membrane and notably improve drought resistance in Arabidopsis [Arabidopsis pathogenesis-related 1, a vital defense protein, has been detected in the extracellular space [The subcellular localization results demonstrated that most of the proteins were extracellular. Corresponding relevant studies have shown that most barley TLPs are located extracellularly, indicating that they may be involved in seed germination . Howeverbidopsis . Arabidoar space . ProteinPH02Gene40419 and PH02Gene31888, which were classified in group 1, indicating that they may have originated from recent gene duplications. These results suggest that the tandem duplication of genes was the primary reason for the expansion of PeTLPs. Similar results have been reported in other studies, and exon-intron patterns in the same phylogenetic classification showed considerable similarity [Further analysis indicated that PeTLPs exhibit certain characteristics in terms of intron/exon patterns, motif structures, and phylogenetic relationships. A total of 26 collinear gene pairs were identified. In addition, several genes with tandem duplications were classified into the same group, such as milarity . Thus, tSequence alignment revealed that PeTLPs contain characteristic cysteine residues and highly conserved REDDD amino acids related to their antifungal activity . SpecifiTwo acidic amino acid residues, Glu and Asp, are considered nucleophiles and proton donors for glycosidic bond cleavage. Glu/Asp-Glu/Asp is the active center of TLPs, with an antiparallel sheet spatial conformation . The 3D A. take. This is consistent with the results of the antifungal activity in a previous protein comparison experiment. A similar set of elements was found in the promoters of Gossypium barbadense TLPs, and different elements in the promoter region of GbTLPs have been suggested to play pivotal roles in abiotic stress and hormonal responses [PH02Gene28874 and PH02Gene28872 contained WRE3 cis-acting elements. Furthermore, PH02Gene34852 and PH02Gene48556 possessed a WUN motif, which was consistent with the transcriptional expression levels of witches’ broom disease in P. edulis. Simultaneously, these genes were significantly upregulated in disease response transcriptional data.The analysis of cis-acting elements revealed that most genes contained domains related to pathogen defense, such as the WRE3 and WUN motif. The cis-acting elements of TLPs demonstrate that these proteins respond to biotic and abiotic stresses . We specesponses . PH02GenPeTLPs were tissue-specific and upregulated . The P. edulis in this experimental base was partially inoculated with A. take two years ago and is now in the peak period of witches’ broom disease. The bamboo in which the witches’ broom symptoms appeared was used as a disease group. The apical buds of white fruiting bodies of A. take with obvious arbuscular symptoms, lateral buds without white fruiting bodies at the lower ends of the arbuscular branches, leaves on the arbuscular branches, and first and second stems at the end of the arbuscular branches were collected in mid-April. Bamboo tissue without arbuscular branches in whole plant was used as a control. All the treatments were conducted in triplicate, and all the samples were stored at −80 °C for RNA-seq and qRT-PCR analysis.In this study, a 5-year-old P. edulis were downloaded from the BambooGDB database (http://www.bamboogdb.org/ (accessed on 1 September 2022)) and saved in cds, genome, gff, and pep formats.The genome and protein sequences of TLP genes, the Hidden Markov Model (HMM) search method was used. HMMSEARCH was used with a threshold e-value < 0.001. Thereafter, we used a species-specific HMM to search for domains, proteins, and cds sequences.To identify http://pfam.xfam.org/ (accessed on 1 September 2022)). The TLP domain sequence was confirmed using the Conserved Domain Database ) and a reliable P. edulis TLP gene sequence was obtained.Sequences obtained from the HMM search were screened and identified according to the definition of the thaumatin domain (ID: PF00314) in the Pfam database (https://web.expasy.org/protparam/ (accessed on 10 September 2022)) [http://cello.life.nctu.edu.tw/ (accessed on 10 September 2022)) [ExPASy (r 2022))  was usedr 2022))  was usedhttp://meme-suite.org/ (accessed on 22 September 2022)) with the default parameter settings of maximum number of motifs = 50 and minimum number of motifs = 10. The corresponding results were shown using TBtools [https://www.ncbi.nlm.nih.gov/cdd (accessed on 22 September 2022)) was used to analyze motif function.PeTLP motifs were identified using the MEME tool ( TBtools . NCBI-CDhttp://gsds.gao-lab.org/ (accessed on 22 September 2022)) according to the P. edulis TLP transcript ID [Exon–intron structures were mapped using the Gene Structure Display Server software ). Finally, we submitted the gene and feature beds to the GSDS website (http://gsds.cbi.pku.edu.cn/ (accessed on 12 October 2022)) to map the cis-acting elements of the genes [We obtained the position information of the he genes .TLP genes in P. edulis using default parameters [PeTLP collinear gene files were uploaded to TBtools software to produce the corresponding collinear circle map.ClustalW was used for multiple sequence alignment to evaluate the evolutionary relationships of the rameters , and Molrameters . The amihttp://www.psc.edu/biomed/genedoc/ (accessed on 2 November 2022)).PeTLP sequences were aligned using MEGA7 software. The results were edited using GeneDoc software  were obtained from NCBI. ClustalW 2.0.10 software was used to compare multiple amino acid sequences between PeTLPs and TLP proteins in other plant species. A phylogenetic tree was constructed using the neighbor-joining method with MEGA 7.0 software. The phylogenetic tree was edited using EvolView (https://www.evolgenius.info (accessed on 2 November 2022)).The TLP amino acid sequences identified in three species ) to determine the gene expression profiles of PeTLPs under different stresses. In the present study, transcriptome data were obtained from the expression levels of leaf PeTLPs under SA, ABA, NaCl, and PEG stress at different time points. Data were normalized to fragments per kilobase of transcript per million mapped reads. To calculate the Log2FC value of genes, clustering [PeTLPs expression levels was generated using the Lianchuan Biological Cloud Platform (https://www.omicstudio.cn/index (accessed on 2 December 2022)).We obtained publicly available transcriptome data from NCBI | > 1 and false discovery rate < 0.05 were the criteria to identify differentially expressed genes (DEGs). A heatmap of transcriptional expression levels of differentially significant expression PeTLPs in P. edulis with witches’ broom disease was generated by Heatmapper and expressed as Log2FC values [Total RNA was extracted using TRIzol reagent  according to the manufacturer’s instructions, RNA samples with high quality were determined using an Agilent 2100 Bioanalyzer system . The Illumina platform was used for transcriptome analysis by employing a 150-bp paired-end library according to the manufacturer’s instructions . Each sample was paired-end sequenced with an Illumina HiSeq 4000 platform. Low-quality reads were filtered. The sequencing data were compared to the reference species C values .https://swissmodel.expasy.org/ (accessed on 12 December 2022)) [https://saves.mbi.ucla.edu/ (accessed on 12 December 2022)) before being used. The corresponding PDB files were downloaded and submitted to the evaluation software, which simultaneously provided six software evaluation results. Three of the results showed whether the model was available. Multiple software packages were used to determine whether the model was good or bad, and Swiss PDB Viewer was used to visualize the protein structure.Gene sequences were selected using the results of transcriptome analysis to construct a 3D protein model on the SWISS-MODEL website (r 2022)) . The genP. edulis samples were stored at −80 °C, and RNA was extracted from the bamboo buds using the commonly used TRIzol method. The presence of RNA was determined by 1% agarose gel electrophoresis. The bands and quantity and purity of the total RNA were analyzed using a Bioanalyzer 2100 and RNA 6000 Nano LabChip Kit . We selected RNA that met the requirements and used the EvoM-MLV Reverse Transcription Kit II  to synthesize complementary deoxyribose nucleic acid (cDNA) from RNA and observed the cDNA bands using agarose gel electrophoresis. The final high-quality cDNA was stored in a −20 °C freezer until quantitative real-time PCR (qRT-PCR) analysis.Glyceraldehyde 3-phosphate dehydrogenase was used as the reference gene [−∆∆CT method [Primer Premier 5 software  was usednce gene . All genT method .TLP genes in the P. edulis genome. PeTLP genes were distributed across 24 scaffolds that were potentially derived from segment duplications during evolution. These genes were classified into three groups and ten subclasses based on the constructed intraspecific and interspecific phylogenetic trees. PeTLP gene promoters were found to contain multiple cis-acting elements associated with hormone and stress responses, which were consistent with the expression pattern of PeTLPs in P. edulis with witches’ broom disease, along with hormone, salt, or drought stress. These PeTLP expression patterns were consistent with the corresponding gene and protein structures. Overall, our findings provide a basis for further functional characterization of PeTLP genes and may be helpful for breeding P. edulis species with disease resistance or enhanced resistance to various abiotic stressors.We identified 81


The incidence and risk of mosquito-borne disease outbreaks in Northwestern Europe has increased over the last few decades. Understanding the underlying environmental drivers of mosquito population dynamics helps to adequately assess mosquito-borne disease risk. While previous studies have focussed primarily on the effects of climatic conditions  and/or local environmental conditions individually, it remains unclear how climatic conditions interact with local environmental factors such as land use and soil type, and how these subsequently affect mosquito abundance.Culex pipiens/torrentium, highly abundant vectors of West Nile virus and Usutu virus. Mosquitoes were sampled at 14 sites throughout the Netherlands. At each site, weekly mosquito collections were carried out between early July and mid-October 2020 and 2021. To assess the effect of the aforementioned environmental factors, we performed a series of generalized linear mixed models and non-parametric statistical tests.Here, we set out to study the interactive effects of land use, soil type and climatic conditions on the abundance of Cx. pipiens/torrentium abundance and sandy rural areas having the lowest. Furthermore, we observed differences in precipitation-mediated effects on Cx. pipiens/torrentium abundance between (peri-)urban and other land uses and soil types. In contrast, effects of temperature on Cx. pipiens/torrentium abundance remain similar between different land use and soil types.Our results show that mosquito abundance and species richness consistently differ among land use- and soil types, with peri-urban areas with peat/clay soils having the highest Our study highlights the importance of both land use and soil type in conjunction with climatic conditions for understanding mosquito abundances. Particularly in relation to rainfall events, land use and soil type has a marked effect on mosquito abundance. These findings underscore the importance of local environmental parameters for studies focusing on predicting or mitigating disease risk.  Mosquitsquitoes . Howeversquitoes . Resultssquitoes . Land ussquitoes . Soil tysquitoes . Unlike squitoes ,33. Howesquitoes ,34. Herehabitats ,35.22.12-baited BG-Pro traps  without additional lures. CO2 was produced following a slightly modified molasses fermentation protocol [Culex pipiens s.l. and its sibling species Cx. torrentium are difficult to distinguish reliably based on morphological characteristics of adult females and were therefore grouped together as Cx. pipiens/torrentium [Mosquitoes were collected at 14 established bird ringing sites distributed across a range of soil types and land uses in the Netherlands . At eachprotocol . A mixtuprotocol . Culex prrentium .Fig. 1Ma2.2Cx. pipiens s.l., the area within a 1500 m buffer around each trapping site was extracted for each land use class  and soil type  using QGIS  [Cx. pipiens/torrentium [Based upon the dispersal-range of m, 2022) . For them, 2022) ,39  . We exclClimate data was taken from the KNMI Data Platform  for each location, trapping week and for twelve time lags, to capture the cumulative legacy effects . Each ti2.3Cx. pipiens/torrentium abundances differ between the six soil/land use classes, we performed a Kruskal-Wallis test followed by Dunn's multiple comparisons post hoc test on the weekly female count data [Mosquito community composition differences between soil types and land uses were analysed by calculating Bray-Curtis similarity distances based on the presence/absence of mosquito species over the entire two-year period ,39. To tunt data .Cx. pipiens/torrentium abundances with a Generalized Linear Mixed Model (GLMM) using a Poisson distribution and log link function [To capture the legacy effects and variation of weekly minimum temperature and precipitation (fig. S4) across all time lags we performed a principal component analysis (fig. S5) . Principfunction . This wafunction . The six3Cx. pipiens/torrentium, Coquillettida richiardii, Culiseta annulata, and Anopheles maculipennis s.l., were present in all soil/land use classes. Most of the collected mosquitoes were Cx. pipiens/torrentium (80%). Mosquito species richness was highest on the non-artificial soils: sandy (14 species) and peat/clay (10 species) compared to artificial soils (4 species). Peri-urban land-uses (10 species) were more diverse than urban land uses (4 species) and were similar to rural land uses: rural low (9 species) and rural high (12 species). There were no trends distinguishable in the Bray-Curtis similarity distance (table S4). The overall community similarity within and between rural land uses across sandy and peat/clay soils is substantial, except between peat/clay rural low and sandy rural high. For further analyses we focused on Cx. pipiens/torrentium mosquitoes, as there were insufficient data for the other species.In total, 9008 adult female mosquitoes from 17 species were trapped at the 14 sampling sites from 2020 to 2021 . The pairwise Dunn's test between soil/land use classes showed that all soil/land uses except peat/clay rural low differed significantly from sandy rural low (Differences in ow areas . There w3.2Cx. pipiens/torrentium abundance. The abundance of Cx. pipiens/torrentium showed a negative relationship to PC1 (i.e. negative to short-term temperature and positive to long-term temperature) for all soil/land use types, with the exception for peat/clay rural-low and sandy rural-low (table S6). The slopes for Cx. pipiens/torrentium abundance in response to minimum temperatures differed significantly between artificial-urban vs. peat/clay-peri urban and artificial-urban vs. sandy-rural high  had a significant interaction with most soil/land use classes in relation to 4Cx. pipiens/torrentium mosquitoes, currently the most abundant and widespread vector species in Northwestern Europe. Interestingly, in our study, the overall highest abundance of Cx. pipiens/torrentium was found in the peat/clay peri-urban class. The site representing this class (UEK) was also the only location where WNV was observed in the Netherlands in 2020 [The aim of this study was to investigate the independent and interactive effects of soil type, land use and climate parameters on the abundance of  in 2020 . HoweverCx. pipiens/torrentium abundances. While temperature effects were similar among land use and soil types, precipitation exhibited a markedly different impact on Cx. pipiens/torrentium abundances, especially in urban and peat/clay peri-urban soil/land use classes.Overall, our results show that most land use and soil types have a significant interaction between temperature, precipitation, and 4.1Cx. pipiens/torrentium abundances between soil/land use classes can partially be explained by Cx. pipiens/torrentium larval habitat preferences in relation to larval habitat availability. As stated earlier, Cx. pipiens s.l. and its sibling species can inhabit a wide range of larval habitats, including artificial habitats [Cx. pipiens/torrentium is low compared to sandy rural areas. This might be due to the lower permeability of peat and clay soils, with clay acting as a barrier on which water can remain and peat acting as a sponge, absorbing excess water via swelling and releasing water as it shrinks [The observed precipitation-mediated differences in habitats ,35. In ghabitats . Additiohabitats . Such tehabitats ,29. Thishabitats . Indeed,habitats . In conthabitats . In rura shrinks . Peat an shrinks ,48,49. S4.2Cx. pipiens/torrentium abundance, regardless of most soil/land use type. However, the relationship between minimum temperature and land use type was not statistically significant for the rural-low land use in both peat/clay and sandy soils. The sampling sites clustered within these soil/land use classes were situated near a large water body, which might play a role in the effects of temperature on Cx. pipiens/torrentium abundances. Overall, we observe that slope directions were similar irrespective of soil/land use type. It is possible that the observed lack of effect of temperature is a result of the sampling design, which involved placing mosquito traps at bird ringing stations located in areas with high woody vegetation. Vegetation can modify microclimatic conditions, including temperature, resulting in a similar environment for mosquitoes across different soil/land use types in terms of minimum temperature. Temperature plays an important role in the physiology, development and survival of mosquitoes and is a strong determining factor for the seasonality in mosquito abundances [We observed a strong relation between temperature and undances ,35. To aundances ,50. Subs5Cx. pipiens/torrentium abundances differently across land use- and soil types. Our findings indicate that the interaction between precipitation and Cx. pipiens/torrentium abundances is influenced by soil type but can be overridden by (peri-)urban land uses, which can alter the availability of artificial breeding habitats. Conversely, the relationship between minimum temperature and Cx. pipiens/torrentium abundances appears to be consistent across land use and soil types. However, our sampling design may have contributed to this result, as our mosquito traps were located in areas with high woody vegetation. This could suggest that patches of high woody vegetation may serve as suitable habitats for adult mosquitoes, irrespective of land use. These results highlight the value of incorporating soil type and land use in conjunction with climatic variables in models to accurately estimate mosquito abundances at higher spatial and temporal resolution than would be possible with climatic variables alone. Overall, our study provides a proof of principle for the importance of soil type and land use for mosquito abundances. As mosquito species differ profoundly in their ecological preferences, particularly in the larval stages, these results cannot be uncritically extrapolated to other mosquito species of medical- and veterinary importance, such as invasive Aedes-species [Cx. pipiens, like Cs. annulata, may react very similarly to differences in land use and soil type. These insights might help to inform management strategies and interventions to mitigate disease risk.Our results imply that precipitation affects -species ,35. HoweDutch Research Council (NWO).This publication is part of the project ‘Preparing for vector-borne virus outbreaks in a changing world: a One Health Approach’ (NWA.1160.1S.210) which is (partly) financed by the https://davidmathlogic.com/colorblind.All figures are made colour-blind inclusive, using the colour-blindness simulation tool of Dr. David Nichols, which can be accessed via Louie Krol: Conceptualization, Investigation, Data curation, Formal analysis, Visualization, Writing – original draft. Rody Blom: Conceptualization, Investigation, Writing – original draft. Martha Dellar: Formal analysis, Writing – review & editing. Jordy G. van der Beek: Validation, Writing – review & editing. Arjan C.J. Stroo: Resources, Writing – review & editing. Peter M. van Bodegom: Supervision, Writing – review & editing. Gertjan W. Geerling: Writing – review & editing. Constantianus J.M. Koenraadt: Conceptualization, Funding acquisition, Supervision, Writing - review & editing. Maarten Schrama: Conceptualization, Writing – review & editing, Supervision, Funding acquisition.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.


Ulcerative colitis (UC) is a chronic inflammatory bowel disease primarily affecting the colon and rectum. Endoscopy plays a crucial role in the diagnosis and management of UC. Recent advancements in endoscopic technology, including chromoendoscopy, confocal laser endomicroscopy, endocytoscopy and the use of artificial intelligence, have revolutionized the assessment and treatment of UC patients. These innovative techniques enable early detection of dysplasia and cancer, more precise characterization of disease extent and severity and more targeted biopsies, leading to improved diagnosis and disease monitoring. Furthermore, these advancements have significant implications for therapeutic decision making, empowering clinicians to carefully consider a range of treatment options, including pharmacological therapies, endoscopic interventions and surgical approaches. In this review, we provide an overview of the latest endoscopic technologies and their applications for diagnosing and monitoring UC. We also discuss their impact on treatment decision making, highlighting the potential benefits and limitations of each technique. Ulcerative colitis (UC) is a chronic inflammatory bowel disease (IBD) characterized by recurrent inflammation of the colon and the rectum . It is aThe chronic inflammatory stimulus in UC patients is associated with a higher risk of developing colorectal cancer (CRC) as compared to the general population . The repEndoscopy plays a crucial role throughout the entire course of the disease, starting from the diagnosis. It is indispensable for obtaining histological confirmation, determining the extent and severity of the disease, monitoring activity, performing dysplasia surveillance and providing endoscopic treatment when possible ,8.In recent years, new technologies have emerged, which could potentially overcome limitations associated with conventional techniques . These aIn this context, the emergence of artificial intelligence (AI) has revolutionized the field of automated image analysis, providing valuable support throughout the management of the disease . By exploring the latest advancements in endoscopic technologies, we will uncover the substantial benefits they offer in terms of accurate diagnosis, effective disease monitoring, risk stratification, prediction of disease outcomes and optimization of treatment strategies .While these advancements offer immense potential, their widespread adoption and integration into routine clinical practice require further validation and refinement . ThroughEndoscopic evaluation plays a critical role in assessing UC activity, by primarily targeting the colonic mucosa and identifying inflammatory changes . The sevTo evaluate disease activity during endoscopy, scoring systems such as ulcerative colitis endoscopic index of severity (UCEIS) and Mayo endoscopic score (MES), rely on these visual characteristics ,15. HoweIn this scenario, AI offers promising solutions by automating the assessment process with increased consistency and objectivity . In the field of AI, machine learning (ML) indicates the development of algorithms that enable computers to learn from data and make predictions or decisions without being explicitly programmed . Deep lep < 0.0001) [Multiple studies have demonstrated that the implementation of AI in UC management can achieve success rates comparable to human experts when using still images. In 2019, a computer-aided diagnosis device (CADx) was developed based on imaging data of endoscopic examinations from 841 patients, exhibiting high discrimination capability between active disease (MES 2–3) and endoscopic remission (MES 0–1), with an impressive area under the curve value (AUC) of 0.98  . Another study replicated comparable outcomes , using a CNN for MES evaluation (MES 0–1 vs. 2–3) and employing over 16,000 images from 3082 patients from the past ten years. Furthermore, the consistency of the MES grade determined by the CNN was found to be equivalent to paired expert reviewer  . This maIn another study conducted by Sutton et al. , the app2 = 77% compared to 82% (75.6–87), I2 = 89% for MES, p = 0.003. Similarly, the sensitivity and positive predictive value of MES were 93.6% (88.7–96.4), I2 = 68% compared to 83.6% (76.8–88.8), I2 = 77% for UCEIS p = 0.007 [In a recent meta-analysis including 12 studies investigating CNN algorithms to assess endoscopic severity in UC, AI trained with UCEIS performed better than MES. The sensitivity and positive predictive value of UCEIS were 93.6% (87.5–96.8), I = 0.007 .Despite advancements in image analysis, a comprehensive assessment of disease severity during endoscopic examinations requires real-time analysis. However, it presents new challenges, such as patient movements, bowel preparation, peristalsis and biopsy-related mucosal injury, which must be accurately recognized and accounted for by the system. In a recent multicenter trial, full-length high-definition (HD) videos were used to train and validate a CNN to predict MES. The results demonstrated agreement rates of 64% for low-definition videos, 78% for HD videos and 82% when compared to central readers . GottlieThe concept of endoscopic remission is expected to evolve towards more in-depth concepts of complete mucosal healing, which indicates the absence of visible inflammation signs and histological healing, which combines both endoscopic and histological remission (HR) ,44. SincAs histological activity is a known predictor of negative clinical outcomes in UC, the implementation of AI may enable the detection of specific endoscopic features that correlate with histology, potentially reducing or eliminating the need for biopsies, which incur additional expenses. Huang et al.  utilizedp < 0.0001), UCEIS  and Robarts histological index (RHI) . A new automated calculation system based on red density technology, which measures image red pixel metrics and vascularity, was developed by Bossuyt et al.  in 2020.Takenaka et al.  developep < 0.001) [In the same group’s subsequent prospective study, the CNN prediction of both endoscopic and HR correlated with significantly better clinical outcomes  . p < 0.001).In an open-label cohort study involving 135 patients with UC in clinical remission, real-time AI-assisted output was used to classify individuals into healing or active group, using the EndoBRAIN-UC  . The priIacucci et al. developed a new AI-based tool for endoscopic assessment called Paddington International Virtual ChromoendoScopy ScOre (PICaSSO); it was used in a prospective study in 2021 that involved more than 1000 videos from 283 patients . Two separate computer models were developed: one trained on WL and the other on virtual chromoendoscopy (VCE) video data. The VCE-AI system performed better than the WL counterpart to detect endoscopic remission (defined as PICaSSO ≤ 3), reaching a sensitivity of 79%, a specificity of 95% and an AUC of 0.94 (95% CI 0.91–0.97). The AI model demonstrated comparable accuracy in predicting HR in both WL and VCE, with accuracies between 80 and 85%. The agreement between the AI system and endoscopist in detecting disease activity was assessed using Cohen’s kappa coefficient. In VCE videos, k was 0.73, 95% CI 0.61–0.85, while in HD-WLE videos, k was 0.51, 95% CI 0.36–0.66. The agreement between the AI system and pathologist in detecting disease activity was assessed in VCE videos (0.45) and in HD-WLE videos (0.59) . In a subsequent study, the PICaSSO system was implemented to detect HR through the PICaSSO histological remission index (PHRI), which utilized an automated dichotomous approach to evaluate neutrophil presence/absence . An AUC PHRI demonstrated strong associations with MES, UCEIS and PICaSSO. Furthermore, a significant correlation was observed between PHRI and clinical outcomes, such as hospitalization, colectomy and therapy modifications triggered by flare-ups, effectively stratifying patients based on their risk within 12 months. Furthermore, a CADx trained with images of 58 patients from a single short wavelength monochromatic LED light illumination endoscope  has been developed by Bossuyt et al. . This inIn addition to artificial intelligence, other endoscopic techniques are gaining ground and may be applied in UC endoscopic assessment. p< 0.0001). In addition, eMHS < 1 may predict sustained clinical remission and a reduced need for hospitalization, steroid therapy and surgery.Confocal laser endomicroscopy (CLE) is an advanced endoscopic technique developed to capture highly detailed and magnified images of the mucosa in the gastrointestinal tract. It relies on the illumination of tissue using a low-power laser, followed by the detection of fluorescent light reflected from the tissue through a pinhole after the application of systemic or topical fluorescence agents . Two difp < 0.05) were registered in UC patients when compared to CD ones. The same parameters where increased in IBD individuals when compared with healthy controls [Another study evaluated 14 morphologic and functional parameters extracted from CLE recordings of 27 UC and 9 control patients to discriminate between IBD and healthy patients and disease activity. A significant increase in the mean intercrypt distance, wall thickness and fluorescein leakage through the mucosa with respective increments of 109%, 117% and 174% (controls . p = 0.02). Furthermore, recently, Vitali et al. highlighted the superior accuracy of endocytoscopic assessment in detecting microscopic disease activity in UC as compared to WLE. A new score, the ELECT (ErLangen Endocytoscopy in ColiTis) score, was developed and validated in 46 UC patients. It showed a strong positive correlation with histopathologic scoring systems (RHI r = 0.70 and NHI 0.73) and superiority over WLE when grading microscopic disease activity  [Another new technique that is taking place in UC evaluation is endocytoscopy (EC) . EC empl= 0.916) . Moreovep < 0.001) [Patients with long-standing UC, excluding those with limited proctitis, have an increased risk of CRC as a result of inflammation-induced carcinogenesis . Recentl< 0.001) . A recen< 0.001) . These r< 0.001) ,62. The p < 0.05) [Ideally, screening and surveillance exams should be conducted on patients in remission by experienced endoscopists skilled in detecting neoplasia associated with IBD. Dysplasia in IBD is often flat and challenging to detect. Initially, the recommended screening method was white light endoscopy (WLE) with random four-quadrant biopsies taken every 10 cm . Standar < 0.05) . Actuall < 0.05) .p = 0.032) in patients with long-standing IBD [p = 0.749) [p = 0.964), but VCE reduced the procedural time by an average of 7 min (p < 0.001) [p = 0.84) [p = 0.617) but less examination time in the Tb group vs. the Rb group  [Another available technology is chromoendoscopy (CE), virtual or dye; the latter involves the application of various dyes to the colon’s epithelium to enhance areas of mucosal irregularity and suspected lesion borders . Dye chrding IBD . However= 0.749) . Nonethe= 0.749) . Recent < 0.001) . An RCT  = 0.84) . Based o = 0.84) . Initial< 0.001) . Althoug< 0.001) .Autofluorescence imaging (AFI) utilizes the properties of light–tissue interactions to enhance neoplastic tissue. When exposed to ultraviolet or short-wavelength visible light, endogenous fluorophores  emit fluorescence . In FINDp = 0.005) [p = 0.015) [p < 0.001) and accuracy , compared to PIT alone [The latest endoscopic techniques, such as confocal laser endomicroscopy (CLE) and endocytoscopy (EC), enable the endoscopist to visualize structures at a subcellular level , obtaining histologic real time in vivo imaging . CLE emi= 0.005) . Moreove= 0.005) . Applica= 0.005) . This fi= 0.005) . EC is a= 0.005) . Kudo’s = 0.005) . In comp= 0.015) . More reIT alone . Althoug® has been successfully developed to distinguish adenoma from malignant lesions observed by EC in non-IBD patients [A CAD system called EndoBrainpatients . In UC ppatients . Nowadaypatients . An evenpatients . They empatients . More stThe emergence of new technologies in endoscopy has led to significant advancements in UC diagnosis and management. In this review, we have highlighted the potential benefits of CE, CLE, EC and AI in improving the assessment and treatment of UC patients. In particular, CE offers improved visualization of subtle changes in the colonic mucosa, allowing for the early detection of dysplasia and cancer . By enhaIn the current era of the treat-to-target approach, endoscopy plays an important role in the management of UC patients. The advanced endoscopic technologies displayed through this review represent a reliable and efficient aid in each step of the course of the disease. Undoubtedly, these tools offer substantial benefits in clinical practice by facilitating standardization during endoscopic exams. Implementation of these technologies can result in significant savings in time, costs, effort and human resources. Although substantial investment might be needed for their integration into daily practice, current data show positive large-scale cost-effectiveness. Moreover, modern endoscopic technologies allow a deeper characterization of the disease, bringing an easier and more tailored therapeutic choice. AI-models may predict UC evolution to avoid potential disease flare-ups in the short-medium term. Soon, AI-based prognostic models will probably encompass the integration of endoscopic, clinical, histological, laboratory and imaging data as well as transcriptional, proteomic and microbiome biomarkers. Despite their promising ability to analyze and integrate plenty of data, currently, these new techniques are not commonly used in clinical practice because of a lack of standardization. It is also relevant to mention that these advanced models offer a prediction rather than unequivocal conclusions, with ethical implications arising in their application. Recently, the first efforts of guidance for clinical trial reporting on AI-model development and clinical usefulness have been taken by the Consolidated Standards of Reporting Trials–Artificial Intelligence (CONSORT-AI) and Standard Protocol Items: Recommendations for Interventional Trials–Artificial Intelligence (SPIRIT-AI) and also by the European Society of Gastrointestinal Endoscopy (ESGE) ,93,94. P


A systematic study on the functional diversity of arsenite methyltransferase from various sources will be used in arsenic bioremediation.Arsenic, a ubiquitous toxic metalloid, is a threat to the survival of all living organisms. Bioaccumulation of arsenic interferes with the normal physiological pathway. To overcome arsenic toxicity, organisms have developed arsenite methyltransferase enzyme, which methylates inorganic arsenite to organic arsenic MMA (III) in the presence of S-adenosylmethionine (SAM). Bacteria-derived Several arsenite methyltransferase protein sequences of bacteria, fungi, fishes, birds, and mammals were retrieved from the UniProt database. In silico physicochemical studies confirmed the acidic, hydrophilic, and thermostable nature of these enzymes. Interkingdom relationships were revealed by performing phylogenetic analysis. Homology modeling was performed by SWISS-MODEL, and that was validated through SAVES-v.6.0. QMEAN values ranged from − 0.93 to − 1.30, ERRAT score (83–96), PROCHECK (88–92%), and other parameters suggested models are statistically significant. MOTIF and PrankWeb discovered several functional motifs and active pockets within the proteins respectively. The STRING database showed protein–protein interaction networks.All of our in silico studies confirmed the fact that arsenite methyltransferase is a cytosolic stable enzyme with conserved sequences over a wide range of organisms. Thus, because of its stable and ubiquitous nature, arsenite methyltransferase could be employed in arsenic bioremediation.The online version contains supplementary material available at 10.1186/s43141-023-00522-9. Arsenic, a ubiquitously found metalloid, has been ranked 20th, 14th, and 12th in the earth’s crust, seawater, and human body respectively based on its occurrence . Differe2−), and hydroxyl radicals (OH), consequently triggering tumor formation  and arsenate [As (V)], enter through aquaglyceroporins and phosphate channels of bacteria respectively and interfere with their metabolism. These elements produce reactive oxygen species (ROS) in the cell, which leads to DNA damage or mutation and impairs enzymatic function , 7. ATP Nowadays, arsenic-resistant bacteria and fungi are being deployed in bioremediation, due to their ability to perform biosorption, bioaccumulation, biotransformation , 9. ThouarsM system has been developed in ancient bacterial strains to detoxify the harmful effects of arsenic. In the presence of SAM (S-adenosylmethionine), arsenite methyltransferase (encoded by arsM gene) can methylate arsenite. Horizontal gene transfer (HGT) plays a key decisive role in the development of resistance across various domains of living organisms  , is common, which transforms monomethylarsonous acid to dimethylarsinic acid [Arsenic became one of the most threatening hazards since early origin of life [about 4 billion years ago (bya)] on earth, and its concentration drastically increased in the late Archean eon 3–2.5 bya). To fight against arsenic toxicity, early organisms developed ArsM enzyme which promoted defense mechanism for respective microorganisms. Gradually, after the Great Oxidation Event , it evolved as an arsenic-detoxifying enzyme . Chen etediation –35. Here.5 bya. Tlic ones . In thisifferent . In the ifferent , and alsifferent , etc. Amnic acid . BesidesArsenite methyltransferase is a thermostable, hydrophilic, evolutionary conserved enzyme involved in arsenite biotransformation . This in silico study sheds some light into the probable role of the amino acids in arsenite methylation. The conserved sequences and domains among different classes of arsenite methyltransferase promote its employability in bioremediation over different ecologies.Additional file 1. List of organisms with protein accession numbers.Additional file 2. Physiobiochemical analysis of enzymes: ExPASy.Additional file 3. Phylogenetic trees of different organisms.Additional file 4. ERRAT values of proteins.Additional file 5. Ramachandran plots of individual organisms.Additional file 6. Chi1-Chi2 scores of selected enzymes regarding individual amino acids.Additional file 7. Ramachandran plots of individual amino acids of selected enzymes.Additional file 8. Structural conservation among the sequences.Additional file 9. Identification of probable signal sequences within enzymes.Additional file 10. Studies on probable transmembrane helixes within enzymes.Additional file 11. Identification of probable motifs.Additional file 12. Results of STRING analysis.Additional file 13. Alignment of most suitable enzymes with the query sequence.


A common complication of anticoagulation therapy is bleeding, especially in patients receiving long-term vitamin K antagonists. Spontaneous intramural hematoma is a rare etiology among life-threatening major bleeds.3, a C-reactive protein of 112.6 mg/L, and a prothrombin time of 75.1 seconds with an international normalized ratio (INR) of 8.5. Abdominal contrast-enhanced computed tomography (CT) showed segmental parietal thickening, luminal narrowing, and partial small bowel obstruction secondary to an intramural jejunum hematoma responsible for a gallbladder occlusion with infiltration of the mesenteric fat in front. The patient recovered two days after conservative treatment.An 80-year-old female patient presented with diffuse abdominal pain. Her history included ischemic heart disease and chronic atrial fibrillation treated with 3 mg of acenocoumarol per day. Three days before her admission, she developed diffuse abdominal pain with fecaloid vomiting, bloating, and not passing gas. Palpation of the abdomen revealed asymmetrical distension and pain, with no signs of peritoneal irritation or bleeding. Investigations showed anemia with a hemoglobin level of 9.2 g/dL, a white blood cell count of 14200/mmIn this case, we report an unusual small bowel intramural hematoma of the jejunum secondary to anticoagulant therapy. Physicians should be aware of this unusual cause of abdominal pain. Early diagnosis may avoid unnecessary surgical exploration. Bleeding is the most common and serious complication of anticoagulation therapy. Approximately 10%-17% of patients receiving long-term vitamin K antagonists will bleed at least once a year .In this case, we report an unusual small bowel intramural hematoma of the jejunum secondary to anticoagulant therapy. Front-line physicians should be aware of this unusual cause of abdominal pain. Early diagnosis may avoid unnecessary surgical exploration. The treatment is conservative in the vast majority of cases .An 80-year-old female patient presented to the emergency unit with diffuse abdominal pain. Her medical history included ischemic heart disease and chronic atrial fibrillation, both treated with 3 mg of acenocoumarol per day. Three days before her admission, she developed diffuse abdominal pain with fecaloid vomiting, bloating, and not passing gas.On admission, her temperature was 37.5 °C, her pulse was 104 beats per minute, her blood pressure was 130/60 mmHg, her respiratory rate was 18 breaths per minute, her oxygen saturation was 96% in ambient air, and she had a Glasgow Coma Scale of 15/15.An abdominal physical examination revealed asymmetrical distension and pain upon palpation but no signs of peritoneal irritation. Similarly, a rectal examination showed no signs of bleeding and no hernia orifices. The rest of the physical examination was normal.The biological assessment showed anemia with a hemoglobin level of 9.2 g/dL, a white blood cell count of 14200/mm3 with 86% segmented neutrophils, a C-reactive protein of 112.6 mg/L, a pro-calcitonin level of 0.43 ng/ml, serum urea nitrogen of 1.38 g/L, serum creatinine of 29.5 mg/L, sodium of 135 mEq/L, potassium of 5.2 mEq/L, and a prothrombin time of 75.1 seconds with an international normalized ratio (INR) of 8.5  showed segmental parietal thickening, luminal narrowing, and partial small bowel obstruction secondary to an intramural jejunum hematoma responsible for a gallbladder occlusion with infiltration of the mesenteric fat in front , and a transfusion of one packed red blood cell. Equally important, we undertook parenteral hydration, nasogastric decompression, discontinuation of acenocoumarol, and digestive bowel resting.The patient recovered two days after conservative treatment. We noted a reduction in abdominal distension and the resumption of transit. On the third day, the patient resumed oral alimentation. Hence, anticoagulation therapy was restarted on the fifth day with an adapted dose. The patient preferred not to use non-vitamin K antagonist oral anticoagulant (NOAC) therapy because of its high price. She left the hospital after seven days.Described in 1838  and repoSpontaneous intramural hematomas of the gastrointestinal tract (SIHGT) are usually observed after abdominal trauma. Many non-traumatic causes can lead to SIHGT, such as anticoagulant therapy, hemophilia, Von Willebrand disease, idiopathic thrombocytopenic purpura, leukemia, lymphoma, vasculitis, collagenosis, chemotherapy, bone marrow transplantation, liver diseases, peptic ulcers, and protein C and S synthesis disorders . Among nSIHGT can present itself through a wide spectrum of symptoms. It can vary from mild, vague abdominal pain to acute abdomen presentation. Common presenting symptoms are abdominal pain, nausea and vomiting, gastrointestinal hemorrhage, and signs of peritoneal irritation or intestinal obstruction ,8.A CT scan is the most commonly performed imaging test for SIHGT diagnosis. A non-contrast CT should be performed before an oral or intravenous contrast CT . In factAn abdominal ultrasound can show a thickened intestinal wall, mainly involving the submucosal layer. This test alone isn’t specific, but when combined with a CT scan, it is efficient ,10.Vitamin K antagonists are widely used drugs around the world. They make up the main therapeutic arsenal for thromboembolic complications of atrial fibrillation with direct-acting oral anticoagulants. Acenocoumarol and warfarin are vitamin K antagonist drugs that prevent the action of vitamin K-dependent factors  by inhibiting gamma-carboxylation. These drugs have a tight therapeutic index and a labile interindividual response. Several foods and drugs can interact with vitamin K antagonists and alter their effects .Patients treated with vitamin K antagonists require frequent INR monitoring. Levels above the therapeutic range increase the risk of bleeding. However, bleeding may also occur in patients with a correct INR target ,13.Shredding of terminal arteries appears to be the most plausible physiopathological explanation of SIHGT. After the vascular rupture, blood leaves the mesentery and infiltrates the intestinal wall muscle. Thus, the hematoma dissects its way between the intestinal mucosa and the muscle layer. Therefore, the viability of the mucosa is preserved, unlike in mesenteric vascular occlusion ,14.All digestive tracts can be affected. Jejunum makes up the most commonly affected part in non-traumatic intramural hematomas of the gastrointestinal tract -16.Medical treatment is preferred in the management of SIHGT unless there are signs of peritonitis. The first step is to discontinue anticoagulant treatment and oral food intake (digestive rest) with nasogastric tube stomach decompression. Vitamin K administration and prothrombin complex concentrates (PCC) can enhance hemostasis . We can The most common side effects of vitamin K antagonists are hemorrhagic accidents. Close monitoring when using those drugs is required to prevent serious hemorrhagic accidents. Intestinal spontaneous intramural hematoma is a rare hemorrhagic accident caused by vitamin K antagonists. Physicians should be aware of this unusual presentation of intestinal obstruction in order to avoid unnecessary surgical exploration.


IntroductionHospital-acquired or nosocomial infections caused by the rapidly emerging bacteria vancomycin-resistant enterococci can be dangerous and even fatal. Therefore, this study aimed to investigate the presence of enterococci in various clinical specimens along with their vancomycin resistance status and biofilm-producing capabilities.MethodsEnterococcus species were isolated and further included in this study. Isolation and identification were done by the standard bacteriological procedure, antibiotic susceptibility testingwas done by clinical laboratory standard guidelines, and biofilm production test was done by microtiter plate methods.A total of 164 ResultsEnterococcus faecalis constituted 60.97% and Enterococcus faecium constituted 39.02%. Maximum isolates were from urine samples. The prevalence of vancomycin-resistant Enterococcus was 6.70%, and 18.29% of Enterococcus isolates were biofilm producers. The sensitivity among the biofilm producers was maximum for linezolid (87.33%), followed by teicoplanin (86.43%) and vancomycin (79.64%).Among the total of 164 isolates, Conclusion biofilm producers Enterococcus isolates were more antibiotic-resistant than non-biofilm producers.High prevalence of enterococci was found in urine samples and Enterococcus cause a wide range of infections in humans. These infections are skin and soft tissue infection, urinary tract infection (UTI), intra-abdominal infection, pelvic infection, bacteremia, meningitis, infective endocarditis, and neonatal sepsis. Enterococci are the third most common agents of nosocomial UTIs. Due to their antibiotic resistance, enterococci are also becoming more and more significant disease-causing organisms in humans, particularly in hospitals [Enterococcus species to commonly used antibiotics including aminoglycosides and beta-lactams, and in actuality, the therapeutic efficacy of clindamycin, trimethoprim/sulfamethoxazole, cephalosporins, and aminoglycosides in controlling Enterococcus is questionable. Significant concern has been raised about enterococci that are vancomycin-resistant and their effects in nosocomial infections. Among the concerns raised, the major one is the emergence of acquired vancomycin resistance in clinical enterococcal isolates [E. faecalis and E. faecium to produce biofilm increases their resistance to antibiotics and increases their capacity to cause disease [Enterococcus species isolated from distinct clinical specimens were assessed for their capacity to form biofilms and their patterns of antibiotic-susceptibility including vancomycin-resistant enterococci (VRE).Enterococci are microbiota of humans and animals. Enterococci live as normal flora of the gastrointestinal (GI) tract, biliary tract, urethra, and vagina. ospitals ,2. Becauospitals . There iisolates ,5. Addit disease . In hosp disease ,8. The c disease ,9. ThereEnterococcus species. Enterococcus species were identified by standard biochemical procedures such as aesculin hydrolysis, acid production from sugars, and PYRase and NH3 production from arginine [The current research was conducted in the Microbiology Department of K. D. Medical College, Mathura, Uttar Pradesh, India. All clinical samples received from January to June 2018 were included in the study, and the study was approved by Institutional Ethics Committee. The inclusion criterion for the current study was that all gram-positive catalase-negative cocci isolated from various clinical samples were considered, and gram-positive cocci  and all gram-negative bacteria were excluded. All clinical specimens  received in the laboratory were cultured for the isolation of arginine . Susceptarginine ,12. PeniDetection of high-level aminoglycoside resistanceEnterococcus was considered susceptible to aminoglycosides, whereas a zone diameter of 6 mm indicated resistance and 7-9 mm indicated that the results were inconclusive [The disc-diffusion method was used to detect high-level aminoglycoside resistance using discs containing high-level gentamicin 120 µg) and high-level streptomycin (300 µg). Plates were incubated, and results were read after 24 hours. When the zone diameter was greater than 10 mm, 0 µg and Detection of minimum inhibitory concentration for vancomycinEnterococcus isolates were lawn cultured, and vancomycin E-strip was placed on it. The minimum inhibitory concentration (MIC) was determined at the point where the ellipse crosses the MIC scale on the strip. As negative and positive controls, E. faecalis ATCC29212 and ATCC51299, respectively, were employed. Results were read into three categories. If MIC came 4 µg/mL or less, it was considered as sensitive, MIC of 8-16 µg/mL was considered as intermediate, and MIC of 32 µg/mL or more was considered as resistant [On Muller-Hinton agar plates, Biofilm formation detectionEnterococci were screened for the biofilm formation by the microtiter plate method. BHI (brain heart infusion) broth with 1% glucose was used as culture medium to detect biofilm. On the basis of optical density reading by an ELISA reader, enterococci were categorized into biofilm non-producers and biofilm producers  -15.Enterococcus species were isolated, of which E. faecalis was 60.97% and E. faecium was 39.02% from various clinical specimens, with p < 0.05 . The Enterococcus were maximum isolated from urine (28.05%), followed by pus (23.78%), and least from body fluids and stool (0.61%), as described in Table Enterococcus species were maximum sensitive for teicoplanin (94.88%), followed by vancomycin (93.38%), linezolid (92.88%), and least sensitive for penicillin (17.19%) followed by norfloxacin (18.75%). Overall, E. faecium were less sensitive to tested antibiotics than E. faecalis, as shown in Table Enterococcus isolated from pus samples were shown maximum (33.33) biofilm-producing capability followed by blood (16.67%) and urine (16.67%); on the other side, bronchoalveolar lavage fluid and pleural fluid sample isolates did not show any biofilm producer, as shown in Table Enterococcus isolates showed good sensitivity to linezolid (87.33%), followed by teicoplanin (86.43%) and vancomycin (79.64%), and least sensitivity to norfloxacin (6.79%). Overall, biofilm-producer isolates showed less sensitivity toward antibiotics compared to non-biofilm producers, as shown in Table Enterococcus showed that males were more infected (72.72%), and the mean age of the patients was 33 years. Enterococcusfaecalis showed more resistance (60.78%) toward vancomycin than E. faecium (39.21%), as shown in Table A total of 164 Enterococcus faecalis was the most common enterococci isolated in our study. These results are concordant with findings of Fernandes and Dhanashree, Jain et al., and Karmarkar et al. [E. faecium was identified as the dominating species by Telkar et al. [E. faecalis was the dominant Enterococcus species, which have been involved in causing a wide range of diseases [Enterococcus species were isolated from urine samples. Different studies from different areas too concluded that urine was the most common sample yielding Enterococci, such as the studies by Mathur et al. [E. faecalis and E. faecium for vancomycin and teicoplanin antibiotics. Additionally, it has been discovered in numerous investigations that although E. faecium makes up a far smaller proportion of clinical enterococcal isolates than E. faecalis, E. faecium was much more resistant to glycopeptides. Less than 2% of E. faecalis were found to be vancomycin-resistant in the research conducted by Deshpande et al. [r et al. -18. In cr et al. . In genediseases . In the diseases , which sdiseases  found thr et al. , Karmarkr et al. , who repe et al. , which we et al. . This ste et al.  revealede et al. . Howevere et al.  reportedEnterococcus prevalence and their drug-resistant status in a particular hospital or area would give a better choice of antibiotics for better disease management and to control drug resistance in enterococci. The prevalence of VRE was found to be 6.7%, and antibiotic resistance was more in biofilm producers compared to non-biofilm producers in the present study. Teicoplanin, linezolid, and chloramphenicol are the drugs that have shown good activity against biofilm-producing and non-producing enterococci; therefore, they can be used in empirical treatment of enterococcal infections.Enterococci are one of the common infectious agents that are capable of causing a wide range of clinical complications. Treating enterococci infection is not that simple compared to other bacterial infections, as it is intrinsically resistant to many commonly used antibiotics such as cephalosporins and aminoglycosides. Moreover, the tendency to acquire drug resistance and the capability to produce biofilm make enterococcal disease management even worse. Hence, the availability of