Proteobacteria was the predominant phylum in all species. Wolbachia was the predominant genus in Haemagogus leucocelaenus. Aedes scapularis, Aedes serratus, Psorophora ferox, and Haemagogus capricornii were the species that showed a greater number of bacterial interactions. Bacterial positive interactions were found in all mosquito species, whereas negative correlations were observed in Hg. leucocelaenus, Ae. scapularis, Ae. serratus, Ps. ferox, and Hg. capricornii. All bacterial interactions with Asaia and Wolbachia were negative in Aedes mosquitoes.Mosquitoes (Diptera: Culicidae) are found widely throughout the world. Several species can transmit pathogens to humans and other vertebrates. Mosquitoes harbor great amounts of bacteria, fungi, and viruses. The bacterial composition of the microbiota of these invertebrates is associated with several factors, such as larval habitat, environment, and species. Yet little is known about bacterial interaction networks in mosquitoes. This study investigates the bacterial communities of eight species of Culicidae collected in Vale do Ribeira (Southeastern São Paulo State) and verifies the bacterial interaction network in these species. Sequences of the 16S rRNA region from 111 mosquito samples were analyzed. Bacterial interaction networks were generated from Spearman correlation values.  The Culicidae family includes 3608 mosquito species (Diptera), classified into 113 genera ,3. MosquAnopheles gambiae and Aedes albopictus larvae reared in the same insectary room and fed on the same food source showed intraspecies and interspecies variations in the composition of the midgut microbiota, concluding that bacterial composition can be modulated by environmental variations, species factors, and complex microbial interactions.Recently, based on the results of an experimental investigation, Saab et al.  found thWolbachia in Aedes aegypti [Asaia in the native microbiome, which inhibits the vertical transmission of Wolbachia [Serratia is present in Anopheles mosquitoes [Aedes mosquitoes, the prevalence of this bacterium is variable. For example, Serratia was more abundant in Ae. albopictus than in Ae. aegypti collected in Houston, TX, United States [Serratia marcescens was found in Ae. albopictus collected in Sing Bur Province of Thailand, and absent in mosquitoes of this species collected in Chumphon and Yala Provinces [Serratia in Ae. aegypti can be low because of the competitive exclusion with Cedecea [Plasmodium also changes the bacterial community [Culex pipiens infected with the West Nile virus showed an increased proportion of Serratia [Spiroplasma sp. PL03 and Weissela cf. viridescens depends on microsporidia infection in the mosquito gut [An. gambiae and Anopheles funestus that were positive and negative groups for Plasmodium falciparum [Asaia borgorensis, Burkholderia fungorum, Burkholderia cepacia and Enterobacter cloacae were present only in females that were negative for P. falciparum. Balaji et al. [Wolbachia can influence the colonization of certain bacterial taxa by competitive interactions, such as the abundance of Serratia sp. in Wolbachia-carrying mosquitoes. The 16S rRNA data have been used to verify microbial co-exclusion/co-occurrence in different organisms [Despite several studies focused on microbial interactions in mosquitoes, it is still unclear how these interactions may contribute to the formation of bacterial communities in mosquitoes. There are few records of natural infection of  aegypti  and anop aegypti ,13. The olbachia ,15. Serrsquitoes ,17, inflsquitoes . In field States ; Serratirovinces . The occ Cedecea . Interacommunity ,23,24,25Serratia . In addiuito gut , and thelciparum . In addii et al.  demonstrrganisms ,28,29. RThe number of studies focusing on bacteria and mosquitoes is increasing. Among these studies are: understanding the acquisition and composition of the microbiota ,20, its This study aims to (1) investigate the microbiota present in eight species of Culicidae collected in Vale do Ribeira, and (2) verify the bacterial communities’ composition and interactions (such as co-exclusion and co-occurrence) between these communities.4H8O2) and immediately preserved in silica gel. Specimens were transported at room temperature to the Laboratório de Entomologia em Saúde Pública–Sistemática Molecular and kept in these conditions until identification and processing. Specimens were morphologically identified using the identification key of Forattini [Adult mosquitoes were collected in a forest preserved area neighboring the town  in Vale do Ribeira, Pariquera-Açu municipality, São Paulo state, Brazil. The collections were performed using an entomological net and conducted daily from 8:00 to 14:30 h from 14 December to 17 December 2021. Mosquitoes were killed with ethyl acetate , 0.3 µm of each primer  [v/v) and purified with Agencourt AMPure XP magnetic beads  according to the manufacturer’s recommendations. After indexing with Nextera XT Index kit (Illumina), the products were purified with magnetic beads and quantified by real-time PCR (qPCR) with the KAPA-KK4824 kit  following the manufacturer’s instructions. All samples were normalised to 4 nM, and an equimolar pool of DNA was prepared. Next-generation sequencing was performed on the Illumina MiSeq sequencer  using the MiSeq Reagent Micro v2 kit (300 cycles: 2 × 150 bp).The mosquito’s surface was rinsed in 70% ethanol and ultrapure water. The genomic DNA of each specimen was extracted separately using the Quick-DNA Fungal/Bacterial Miniprep kit (ZymoReasearch), following the manufacturer’s instructions. The V4 region of the 16S rRNA gene was amplified using isolated DNA from each specimen. Each reaction was carried out in a final volume of 20 µL containing 1 X GoTaqTAAT 3′) , 8 µL ofFLASH v. 1.2.11  was usedqiime tools import command and Casava 1.8 single-end demultiplexed format were used to import the joined sequences to QIIME2. Quality control and denoising were performed with the commands: qiime quality-filter q-score and qiime deblur denoise-16S, respectively. To taxonomic attribution was used SILVA 138 [qiime feature-classifier classify-sklearn.The ILVA 138 ,36 and qqiime diversity α-rarefaction command to construct this curve.The rarefaction curve was generated to obtain the expected number of ASVs (Amplicon Sequence Variants) in each sample for a given number of sequences, and to allow for the comparison between the richness of the samples. We used a depth value of 10,000 and qiime diversity core-metrics-phylogenetic, and a phylogenetic tree was constructed with qiime phylogeny align-to-tree-mafft-fasttree.Diversity metrics (α and β) were generated with Shannon-Weaver indices (α diversity) were subject to Kruskal-Wallis followed by Dunn’s test, adjusted with Bonferroni method in RStudio v.1.4.1106 to verify whether samples from one species have greater α diversity than from other species.Analysis of variance (PERMANOVA) statistical test was performed with β diversity data generated by Unifrac weighted and unweighted distances in QIIME 2. This statistical test makes it possible to verify whether bacterial diversity differs significantly between mosquitoes of different species.qiime taxa collapse, qiime composition add-pseudocount, and qiime composition ancom. This study makes it possible to verify whether any ASV is more abundant in a particular mosquito species than in another.Microbiome composition analysis (ANCOM) was performed in QIIME 2 using ASV table, SILVA taxonomy and the commands: Data of the weighted and unweighted Unifrac phylogenetic distance matrices generated in QIIME2 were used to perform Principal Coordinate Analysis (PCoA) in RStudio v.1.4.1106. PCoA images were generated with tidyverse and qiime2R packages available in RStudio. These images allow for visualizing the distance between the bacterial communities of each sample. Heatmaps were obtained in RStudio v.1.4.1106 employing data of ASV table and taxonomy. The images generated allow for verifying the abundance of each taxon per sample.The ASV table was changed as described in Hegde et al.  and was p ≥ 0.05. The non-discarded data were used to infer a bacterial interaction network, with the blue lines corresponding to a positive correlation, whereas red lines corresponded to a negative.After data normalization, a Spearman correlation matrix was generated. This matrix and analysis were performed for each species separately and carried out with igraph, Hmisc and Matrix packages available in RStudio. Spearman correlations with values of r ≤ 0.75 were discarded, as well as with Ae. scapularis (08), Ae. serratus (06), Hg. capricornii (07), Hg. leucocelaenus (14), Ke. cruzii (15), Ps. ferox (12), Sa. conditus (30) and Wy. confusa (19)  .A total of 9,647,379 (R1 or R2) raw reads were generated in the NGS. These reads varied between 58,143 and 123,070 in the samples . After jAe. scapularis showed 530 ASVs; Hg. leucocelaenus, 310; Hg. capricornii, 334; Ps. ferox, 874; Ae. serratus, 504; Sa. conditus, 1014; Wy. confusa, 623; and Ke. cruzii, 546 , and then the Kruskal–Wallis test  followed by Dunn’s test adjusted with the Bonferroni method was performed to know which species differed in α diversity. The following groups showed significant differences in the Shannon indices: Hg. leucocelaenus–Ke. cruzii (p = 0.002); Hg. leucocelaenus–Sa. conditus (p = 0.002) and Hg. leucocelaenus–Wy. confusa (p = 0.026).Shannon-Weaver indices ranged from 0.607 to 6.355 between samples . These iAe. scapularis and Hg. leucocelaenus, Ke. cruzii, Sa. conditus, and Wy. confusa; (2) Ae. serratus and Hg. capricornii, Hg. leucocelaenus, Ke. cruzii, Sa. conditus, and Wy. confusa; (3) Hg. capricornii and Ke. cruzii, Sa. conditus and Wy. confusa; (4) Hg. leucocelaenus and Ke. cruzii, Sa. conditus, Wy. confusa, and Ps. ferox; (5) Ke. cruzii and Sa. conditus and Ps. ferox; (6) Ps. ferox and Sa. conditus and Wy. confusa; and (7) Sa. conditus and Wy. confusa , Venezuelan Equine encephalitis, and Wuchereria bancrofti [Ae. serratus is considered a secondary vector of both the Ilheus virus [Hg. leucocelaenus is the primary vector of sylvatic YFV in the New World and was involved in Brazil’s greatest yellow fever outbreak between 2016 and 2018 [Ke. cruzii is the primary vector of Plasmodium sp. in Atlantic Forest  [Ps. ferox has already been found to be naturally infected with the Rocio virus in Vale do Ribeira region [Wy. confusa from Capivari-Monos EPA  was infected with the Zika virus [Because the mosquito species analysed in this study have great public health importance as potential vectors of arboviruses, the knowledge of bacterial communities present in these insects is essential for further investigating the bacteria-pathogen interactions and their influence on the ability of these species to be infected and transmit parasites. ancrofti ,43,44. Aus virus  and the us virus ,38. Hg. malaria) ,47, and malaria) . Ps. fera region , with YFa region . Wy. conka virus .Many factors can modulate mosquitoes’ bacterial composition, including the mosquito species  and compHg. leucocelaenus and Ke. cruzii and Sa. conditus and Wy. confusa. Permanova analyses showed that most mosquito species studied differed in their bacterial communities. These results corroborate other studies in the literature [Ke. cruzii linked to bromeliad phytotelmata; Wy. confusa, Sabethes, and Haemagogus with phytotelmata provided by bamboo and tree holes; and other species are relations with temporary ground pools, such as Ae. scapularis, Ae. serratus and Ps. ferox. Although all specimens have been collected in the same macroregion, the larval habitats (microregions) are ecologically distinct. As bacterial communities present in immature habitats can contribute to bacterial composition in adult mosquitoes [Statistical tests showed a significant impact of the species on the α diversity of the mosquito, with an effect verified between terature ,51,52, asquitoes , the bacAe. serratus showed the highest interactions between bacterial communities, followed by Hg. capricornii, Ae. scapularis, and Ps. ferox.There are several interactions between microorganisms, such as mutualism, competition, and commensalism . IncreasAfipia was found in all mosquito species, being found predominantly in Wy. confusa, Ke. cruzii, and Sa. conditus. Despite being in lower abundance in Ae. scapularis, Ae. serratus, Hg. capricornii, and Ps. ferox, in these species, Afipia interacted with at least six other bacterial genera. Afipia is a Gram-negative rod bacterium of the phylum Proteobacteria. The genus was described in 1991 [Afipia felix) appears to be related to cat scratch disease [Afipia in mosquitoes. in 1991 , and one disease . To dateAcinetobacter was found in all mosquito species studied. Species of this genus may be involved in blood digestion and parasite–vector interactions in Ae. albopictus [Delftia were found in females of Nyssorhynchus darlingi not infected with Plasmodium [Delftia and Asaia in Aedes mosquitoes. The negative interaction between these bacterial groups needs further investigation to verify if the co-exclusion hypothesis of these bacterial genera can occur in other Aedes mosquitoes.bopictus . Bacteriasmodium  in otherasmodium . The bacAsaia is frequently found in Aedes and Anopheles mosquitoes [Plasmodium in female mosquitoes [Asaia can diminish the longevity of infected males of Anopheles stephensi [Asaia and Wolbachia showed only negative interactions with other bacterial genera in the Aedes samples analyzed in this study. Our findings corroborate the results of an investigation to verify the occurrence of reciprocal negative interference between these bacterial groups to colonise the gonads [Aedes species from Vale do Ribeira. Few Wolbachia sequence readings were found in Ae. scapularis and Ae. serratus, while Asaia readings were more abundant in both species. Although the Wolbachia-Asaia mutual exclusion hypothesis has been found in Hg. leucocelaenus specimens from Vale do Ribeira [squitoes ,61, and squitoes . In additephensi . It is ne gonads . Althoug Ribeira , the samWolbachia infection in Ae. aegypti causes upregulation in the transcription of genes related to reduction–oxidation reactions and immunity. This leads to an increase of reactive oxygen species and induction of oxidative stress in the host and showing that to favor infection, the bacterium can manipulate the host’s defense system [e system .Wolbachia strains induce cytoplasmic incompatibility (CI), resulting in lethality of embryos generated from females not infected with Wolbachia or infected with a different strain present in the male [Ae. albopictus [Ae. aegypti [Hg. leucocelaenus and this species of mosquito is considered the primary vector of sylvatic yellow fever virus in the south-eastern region of Brazil [Wolbachia strains present in these mosquitoes, and to know if the existing strains can harm the spread of the yellow fever virus.Many of the the male . It manithe male . In addibopictus  and to i aegypti . Thus, cf Brazil , furtherSerratia bacteria can modulate the vector competence of populations of Ae. aegypti for the Zika virus by interfering with mosquito salivation [Serratia 16S readings were found in one female of each species, Sa. conditus and Ps. ferox. The competitive exclusion between Serratia and Cedecea was verified in Ae. aegypti co-infected with these bacteria [Cedecea was recovered from the specimens of the species of the Aedini tribe from Vale do Ribeira. Consequently, the absence of Serratia in Aedes and Anopheles mosquitoes cannot be explained by the exclusion effect between Serratia and Cedecea symbionts. It is likely that other bacteria can be responsible for the competitive exclusion with Serratia.livation  and intelivation . In thisbacteria . No readOur findings uncovered that the analyzed species exhibited clear differences in their microbiota composition based on the α and β diversity indices. Furthermore, some bacterial interactions in this study corroborated previous findings in the literature, while other interactions will need further investigation. In addition, it will be important to verify if the interactions are observed in different spatial and temporal scales in subpopulations of a mosquito species.Wolbachia and Asaia are predominant in species of the Aedini tribe, such as Hg. leucocelaenus and Aedes mosquitoes. The bacterial interaction network was verified for each mosquito species showing that Shingobium, Unclassified Pseudonocardiaceae, Afipia, Delftia, and Shingomonas had more bacterial interactions in the Aedes specimens. Bacterial interactions with Asaia and with Wolbachia were negative in Aedes mosquitoes.Here we uncover variations in microbial composition among different Culicidae species.


Genome-scale metabolic reconstruction tools have been developed in the last decades. They have helped to reconstruct eukaryotic and prokaryotic metabolic models, which have contributed to fields, e.g., genetic engineering, drug discovery, prediction of phenotypes, and other model-driven discoveries. However, the use of these programs requires a high level of bioinformatic skills. Moreover, the functionalities required to build models are scattered throughout multiple tools, requiring knowledge and experience for utilizing several tools.Here we present ChiMera, which combines tools used for model reconstruction, prediction, and visualization. ChiMera uses CarveMe in the reconstruction module, generating a gap-filled draft reconstruction able to produce growth predictions using flux balance analysis for gram-positive and gram-negative bacteria. ChiMera also contains two modules for metabolic network visualization. The first module generates maps for the most important pathways, e.g., glycolysis, nucleotides and amino acids biosynthesis, fatty acid oxidation and biosynthesis and core-metabolism. The second module produces a genome-wide metabolic map, which can be used to retrieve KEGG pathway information for each compound in the model. A module to investigate gene essentiality and knockout is also present.Overall, ChiMera uses automation algorithms to combine a variety of tools to automatically perform model creation, gap-filling, flux balance analysis (FBA), and metabolic network visualization. ChiMera models readily provide metabolic insights that can aid genetic engineering projects, prediction of phenotypes, and model-driven discoveries.The online version contains supplementary material available at 10.1186/s12859-022-05056-4. Genome-scale metabolic reconstructions (GSMRs) are essential tools in system biology . Over thRecently, several tools such as AureMe , PathwayHere we present a novel tool named ChiMera, which compiles widely used tools for genome-scale metabolic modeling in a single pipeline. ChiMera uses a protein sequence as input (*.faa file) and creates a model based on a highly curated universal model . The resChiMera uses automation algorithms to combine three main steps in GSMR, i.e. model creation and gap-filling, FBA, and pathway visualization. The tool also includes a submodule that enables users to perform in silico gene and reaction essentiality screening based on FBA. All these functions are modular and compatible with further expansions of ChiMera. Fig. .Fig. 2FlWe use CarveMe (v1.5.1) in the reconstruction module of ChiMera. The initial draft model is created based on the protein sequence file (*.faa file) provided by the user. During the reconstruction process, ChiMera uses the CarveMe gap-filling algorithm to add missing reactions based on a given growth media. This process uses genomic evidence to ensure that the model will be able to predict growth in the condition. CarveMe uses a top-down approach in a pre-built reference and manually curated universal model . It applCarveMe utilizes five predefined media: LB, anaerobic LB, M9, anaerobic M9, and M9 using glycerol as a carbon source . A tab-sWe used COBRApy (v0.22.1) to convert the initial draft into an S-matrix and perform FBA analysis . Growth,ChiMera converts the initial SBML model to 2 different model formats: JSON, and YAML. These model formats are compatible with the majority of currently available tools.in-house algorithms to automate the generation of metabolic maps based on Escher (v1.7.3) [We performed transformations in the JSON model to enable compatibility between Escher maps and the user model. We developed (v1.7.3) . Ten pre(v1.7.3) , 15. TheFurthermore, we developed a second visualization module that creates files compatible with graphical visualization tools. ChiMera automated the use of PSAMM (v1.1.2), converting the model to a graphical representation . The graPseudomonas putida KT2440 and Escherichia coli) and one Gram-positive bacterium (Bacillus subtilis). Protein sequence files were downloaded from NCBI under the accession numbers NC_002947.4, NZ_CP020543.1, and AL009126.3. These genomes were used to generate the models, visualizations and knockouts. Further, these ChiMera models were compared with manually curated models from the BiGG database, iJN1463 (P. putida), iEC1344_C (E. coli), and iYO844 (B. subtilis).For demonstrating functionality of ChiMera, we selected two well-studied Gram-negative bacteria  and reactions (edges).Next, the organism-specific model is automatically converted to a S-matrix, using COBRApy. The biomass-producing reaction, which contains the precursors like carbohydrate, protein, lipids, and energy molecules balanced for producing one gram of biomass, is set as the biological objective function for performing FBA Fig. C. The flThe output of PSAMM can be directly loaded into Cytoscape , produciChiMera also produces a broader view of the target metabolism Fig. E. The coTo allow ChiMera's flexibility and modularity, users can also provide a pre-built model with the protein sequence file, which should hold the same prefix, directly performing FBA analysis and construction of the pathway maps. Documentation is provided to ensure that the annotations of the model or the presence of extra compartments are compatible with PSAMM, to generate the Cytoscape compatible file. We provide tutorials on how to use ChiMera output files to build custom maps for any organism .The knockout module of ChiMera is dependent on COBRApy . Here, wP. putida) and iJN1463 models shared 68% of their metabolites and 60% of their reactions. Similar values were observed for iChiMera1657 (E. coli) and iEC1344_C models. iChiMera1182 (B. subtilis) shared 50% of its metabolites and 44% of its reactions with iYO844 : Mac Os, Linux.Programming language: Python.Other requirements: The tool has dependence on other thirst party software. All dependencies are handled during the installation and creation on an virtual environment.License: GNU GLP 3.Any restrictions to use by non-academic: ChiMera has no restriction, however CarveMe rely on IBM ILOG CPLEX Optimization Studio, that demands an academic license.Additional file 1. Figs. S1–S7 ChiMera output examples and supplementary analysis.Additional file 2. Tables S1–S7 ChiMera supplementary tables and metadata.


Background: The increase in extended-spectrum β-lactamase (ESBL) producing microbes in recent years represents a major challenge.Escherichia coli in patients requiring hospitalization for treatment.Aim: To study the risk factors for urinary tract infections (UTIs) caused by ESBL-producing E. coli strain cultured on urine culture between January 1 and December 31, 2018. The electronic health care records of these patients were searched to identify those patients with previous UTIs due to an ESBL-producing E. coli grown on urine culture. Patients with cancer or those taking prophylactic antibiotics or immunosuppression were excluded.Materials and method: Electronic health records were used to identify 616 inpatients over the age of 18 who had UTI symptoms and/or signs and an ESBL-producing E. coli included male sex (P = 0.0032), age over 66 years (P < 0.0001), renal stones (P = 0.0021), urology intervention within six months of presentation (P = 0.0360), pressure sores (P = 0.0002), feeding tubes (P = 0.0076), and urinary catheter (P = 0.0023). Comorbidities .Result: Risk factors for the acquisition of ESBL-producing E. coli. However, the recognition of risk factors for UTI caused by ESBL-producing E. coli may facilitate the early detection of high-risk cases and guide treatment decisions. This can improve patient outcomes while decreasing the length of the hospital stay.Conclusion: Antimicrobial stewardship programs may have reduced the development of antimicrobial resistance in  Escherichia coli is the most frequently identified microorganism.Urinary tract infection (UTI) is the second most common cause of community-acquired infection and healthcare-associated infection in clinical practice worldwide ,2. The pBacteria can evolve highly specialized mechanisms of resistance to commonly used antibiotics . One resExpression of an ESBL confers resistance to many of the first-line antibiotics commonly used to treat UTIs. These include penicillin, third-generation cephalosporins , and monobactams  ,4. TreatE. coli was conducted at King Khalid University Hospital, Riyadh, Saudi Arabia, from June 2009 to June 2011  with recurrent UTI due to ESBL-producing E. coli. The demographic profile, comorbidities, antibiotic use, and imaging of the study population are summarized in Table At KAMC in 2018, symptomatic UTI caused by ESBL-producing Demographic dataE. coli UTI (61.90%) than those 66 years old or less (P < 0.0001). Male patients (39.88%) were also at higher risk of recurrent ESBL-E. coli UTIs (P = 0.0032).Approximately half of the study population  were over 66 years old at presentation to the hospital. This subgroup was at greater risk of recurrent ESBL E. coli UTI Comorbid disease, indwelling urinary catheters and risk of recurrent ESBL E. coli was not significantly affected by diabetes mellitus (P = 0.4680), hypertension (P = 0.3633), chronic kidney disease (P = 0.5818), stroke (P = 0.1244), or prostate disease (P = 0.1303). However, the risk of recurrent UTI with ESBL-producing E. coli was significantly increased by renal stones (P = 0.0021), pressure sores (P = 0.0002), tube feeding (P = 0.0076), and urology intervention within the six months prior to presentation (P = 0.0360).The risk of recurrent UTI with ESBL-producing E. coli UTU was increased in patients with an indwelling urinary catheter (P = 0.0023). The risk of recurrence was greater in those patients who had required urinary catheterization for more than one month (P = 0.0014). However, there was no difference between intermittent catheterization and the use of a permanent catheter (P = 0.4668).The risk of recurrent ESBL-E. coli (P = 0.0001). However, the findings of renal tract ultrasound  were not statistically significant (P = 0.0752).Renal ultrasound was more likely to have been performed in those patients who had recurrent UTI caused by ESBL-producing E. coli UTIAntibiotic therapy and risk of recurrent ESBL E. coli was not affected by antibiotic use (P = 0.3826) or the duration of antibiotic use . The specific choice of antibiotic  did not affect recurrence (P = 0.0689).The risk of recurrent UTI due to ESBL-producing E. coli was higher in patients receiving intravenous antibiotics than that in patients treated with an oral antibiotic (P = 0.0265).However, the risk of recurrence was influenced by the frequency of antibiotics (P = 0.0004). The risk was higher in those who received antibiotic therapy twice a day (BID) or three times a day (TID). The risk of recurrence was less in patients who received antibiotics daily or four times a day (QID). The risk of recurrent UTI caused by ESBL-producing E. coli. It is therefore important to determine the risk factors for these multidrug-resistant organisms.The current paradigm for antibiotic stewardship in the treatment of UTI prioritizes the use of first-line therapies such as trimethoprim and nitrofurantoin . HoweverE. coli UTIRisk factors for recurrent ESBL E. coli UTIs were identified in the present cohort. These were men over the age of 66 who had had urology intervention within the previous six months, such as renal stones, pressure sores, feeding tubes, or a urinary catheter. These findings are consistent with previous studies.Seven risk factors for recurrent ESBL Age and Male Sex E. coli [Age and male sex are well-recognized risk factors for UTI due to ESBL-producing E. coli -8. IncreE. coli . FurtherUrinary Tract Catheterization and Other Urological InterventionsE. coli UTI [E. coli UTI than if the catheter had been present for less than one month. Urinary catheters may become coated with a biofilm that acts as a reservoir for microorganisms [Catheterization of the urinary tract increases the risk of ESBL coli UTI -12. The rganisms .E. coli in the present cohort. Further studies are required to define the selection of peri-procedural antibiotic prophylaxis in patients at risk of ESBL-producing E. coli who undergo urological intervention.Similar to catheterization, other interventions in the urinary tract also compromise the action of antibiotics and host defense mechanisms . Thus, cRenal Ultrasound, Renal Disease and CalculiE. coli. While the findings of the renal ultrasound did not correlate with risk, the risk of recurrent UTI due to ESBL-producing E. coli was increased in patients with renal stones but not chronic kidney disease. In contrast, Al-Otaibi et al. [E. coli were more common in patients with renal disease (P = 0.017) but less common in patients with renal stones [Patients in the present cohort who had a renal ultrasound prior to presentation were more likely to have a recurrent UTI with an ESBL-producing i et al.  reportede P = 0.07 but lesE. coli infections [Renal calculi increase the risk of infection due to ESBL-producing GNB . Urinaryfections . Ureterifections . Stasis fections . Urinaryfections  may reflPressure Sores and Tube FeedingE. coli. This finding is consistent with previous reports [Tube feeding and the presence of pressure sores were also associated with a significantly increased risk of recurrent UTIs with ESBL-producing  reports ,14. The E. coli UTIFactors that did not increase risk of recurrent ESBL E. coli.When considering empirical antibiotic therapy for UTI prior to the availability of culture results, it is important to determine the risk of ESBL-producing GNB. It is therefore important to determine the factors that do not increase the risk of ESBL-producing Diabetes MellitusE. coli was significantly higher in patients with diabetes (P=0.001). However, in the present cohort, the risk of recurrent UTI due to ESBL-producing E. coli was not affected by diabetes. Previous studies have also reported similar results [Aswan et al. reported that the prevalence of ESBL-producing es P=0.00. However results ,17. ThisIt is important to note that none of the patients in the present cohort were treated with sodium glucose-like transporter 2 (SGLT-2) inhibitors. Some studies suggest that the risk of UTI is increased by these agents . FurtherHypertension E. coli. While previous studies have reported similar findings [E. coli [E. coli, hypertension may have been a confounder in previous studies.In the present cohort, hypertension was not associated with an increased risk of UTI due to ESBL-producing findings , the dat[E. coli . HoweverProstate DiseaseE. coli. While previous studies have reported similar findings [E. coli [In the present cohort, prostate disease was not associated with UTI due to ESBL-producing findings , data on[E. coli .As described above, urethral catheterization and urological procedures increase the risk of UTI -14. The Prior Antibiotic Use E. coli [E. coli. Studies published in 2014 and 2016 reported similar findings [E. coli [Prior antibiotic use is the most frequently identified risk factor for UTI due to ESBL-producing E. coli ,22. Howefindings ,22. Antifindings . This ap[E. coli .LimitationsE. coli. This may be because only inpatients were included in the present cohort. Thus, it is not possible to determine whether a hospital stay or antibiotic treatment affected the risk of ESBL-producing E. coli. In addition, the data for our study were collected from a single-center study; therefore, this study’s findings cannot be generalized to the regional or national level.The limitations of the present study include being a retrospective study and the lack of a control group. Thus, potential confounding factors may have influenced statistical analyses. Furthermore, the present study did not detect any association between specific antibiotic usage and ESBL-producing E. coli in a tertiary care center in Riyadh, Saudi Arabia. Male sex, age over 66 years, renal stones, urology intervention within six months of presentation, pressure sores, feeding tubes, and a urinary catheter were associated with an increased risk of UTI due to ESBL-producing E. coli, while comorbidities  and the duration of antibiotic therapy were not associated with an increased risk of recurrence of an ESBL-producing E. coli UTI. Antimicrobial stewardship programs may have reduced the incidence of antibiotic resistance in E. coli. However, the identification of patients at high risk of UTI due to ESBL-producing E. coli is required to guide the choice of empirical therapy.The prevalence of ESBL infection in patients and healthcare professionals is influenced by several risk factors. Most studies agree that factors such as hospital admission and previous antibiotic use are important risk factors. However, the relevance of other factors  is less clear. The present study identified risk factors for UTI due to ESBL-producing


Aotus), or “night monkeys” are platyrrhine primates in the Aotidae family. Early taxonomy only recognized one species, Aotus trivirgatus, until 1983, when Hershkovitz proposed nine unique species designations, classified into red-necked and gray-necked species groups based predominately on pelage coloration. Recent studies questioned this conventional separation of the genus and proposed designations based on the geographical location of wild populations. Alu retrotransposons are a class of mobile element insertion (MEI) widely used to study primate phylogenetics. A scaffold-level genome assembly for one Aotus species, Aotus nancymaae [Anan_2.0], facilitated large-scale ascertainment of nearly 2000 young lineage-specific Alu insertions. This study provides candidate oligonucleotides for locus-specific PCR assays for over 1350 of these elements. For 314 Alu elements across four taxa with multiple specimens, PCR analyses identified 159 insertion polymorphisms, including 21 grouping A. nancymaae and Aotus azarae (red-necked species) as sister taxa, with Aotus vociferans and A. trivirgatus (gray-necked) being more basal. DNA sequencing identified five novel Alu elements from three different taxa. The Alu datasets reported in this study will assist in species identification and provide a valuable resource for Aotus phylogenetics, population genetics and conservation strategies when applied to wild populations.Owl monkeys (genus  Aotus spp.) are small, neotropical platyrrhine primates (monkeys found in the Americas) within the Aotidae family; they are often referred to as night monkeys and are widespread across South America and Panama . The purpose of this study was to computationally ascertain a dataset of the youngest Alu insertions specific to the owl monkey lineage from the [Anan_2.0] genome and to perform locus-specific PCR on a DNA panel of 23 captive Aotus specimens to identify Alu insertion polymorphisms. The Alu datasets reported in this study provide a valuable resource to assist in species identification and to facilitate future studies of Aotus phylogeny, population genetics and conservation strategies.The first platyrrhine-specific elements . This ple family . Other pmarmoset , squirremarmoset , capuchimarmoset ,44 and omarmoset ,45. Somehylogeny ,29,46,47Alu insertions from the A. nancymaae genome [Anan_2.0] was performed as described previously  genome; or (3) polymorphic pattern for insertion presence/absence (N = 149). These results correspond to a ~50% polymorphism rate within the candidate dataset. These 149 insertion polymorphisms were further divided into four general categories. (1) The target Alu insertion was restricted to the reference genome [Anan_2.0] species A. nancymaae and was homozygous present (h.p.) in all A. nancymaae samples (PCR code: A.n. (h.p.)) N = 31. An example is shown in A. nancymaae species identification. (2) The target Alu insertion was restricted to A. nancymaae and was polymorphic for insertion presence/absence among the nine A. nancymaae samples on our DNA panel (PCR code: A.n. (poly)) N = 83 (Alu insertion polymorphisms could be applied to the study of population genetics among A. nancymaae wild populations. (3) The target Alu insertion was present in both A. nancymaae and A. azarae samples while being absent in A. trivirgatus and A. vociferans (PCR code: A.n. and A.a.) N = 21 (Aotus phylogeny. (4) The target Alu was homozygous present in A. nancymaae, A. azarae and A. trivirgatus while being absent from A. vociferans, N = 3   with >2%er 2022) . Each lofamilies , summari) N = 83 B. These ) N = 21 C. These s, N = 3 D, suggesAotus samples, with the exception of two A. vociferans individuals who lacked the insertion (Alu insertion polymorphisms (Aotus_127 and Aotus_1602) supported an alternative topology in which the target Alu was present in A. nancymaae, A. azarae and A. vociferans while being absent from A. trivirgatus . Future investigations with expanded species sampling and sample sizes are required to determine the utility of these novel elements. Sequencing also identified traces of non-repeat DNA with the potential for species identification.All potential parsimony informative insertion polymorphisms, as well as any ambiguous PCR fragments or evidence of incomplete lineage sorting (ILS), were subjected to DNA sequencing. These data are shown as Alu insertion polymorphisms shared by A. nancymaae and A. azarae to the exclusion of A. trivirgatus and A. vociferans. DNA sequencing confirmed 19 of these (Aotus_1124). DNA sequencing of the remaining two candidates revealed the existence of “near parallel insertions”, a different Alu insertion from the intended target, but still located between the oligonucleotide primers producing a PCR amplicon of the predicted “Alu-present” size. The PCR-based gel image for Locus Aotus_404 is shown in A. nancymaae and A. azarae, while the sequencing alignment  but also revealed that A. vociferans 86218 has a different Alu in the opposite orientation about 115 bp upstream of the target  was designed that could be used in conjunction with the existing reverse primer to potentially genotype this newly discovered A. trivirgatus derived Alu element separately from the [Anan_2.0] ascertained target insertion  are unrelated males._2LS_078  and Locutus_1518 . In addiividuals . DNA seqividuals , placingAotus_1288V2 .For Locus s_1288V2 A, the taenotypes . This loAlu element, in addition to the target insertion, that was shared by A. trivirgatus and A. vociferans to the exclusion of A. nancymaae and A. azarae .Locus Owl_2LS_042 was originally suspected of having a second . azarae ; if confciferans . The tarA. vociferans as the most basal Aotus species, we also recovered three other Alu insertions  in addition to the intended target insertion  and 314 characters to perform a maximum parsimony heuristic search. Of the 314 Alu insertion loci, 165 were constant, and another 114 were restricted to A. nancymaae, leaving 35 potentially parsimony informative among Aotus species   and are, therefore, not subject to the same level of directional ascertainment bias. Until more genomes for other Aotus species become available, the five novel Alu elements reported here provide alternate evolutionary perspectives to [Anan_2.0].The number of r humans  and somer humans ,44, a feAlu insertion polymorphisms were identified from 37 different platyrrhine Alu subfamilies, all of which were AluTa15 derived , squirrel monkey, [SaiBol1] and capuchin, [Cebus imitator_1.0]. This strategic approach facilitates the discovery of the youngest Alu subfamilies with the ongoing proliferation of progeny elements. derived . The higAotus lineage-specific Alu elements and demonstrates their utility as phylogenetically diagnostic insertion polymorphisms. It shows that multiple Alu subfamilies have evidence of recent mobilization within owl monkey genomes. Alu elements restricted to A. nancymaae, as well as the discovery of novel Alu elements from three additional owl monkey taxa, will assist in species identification. Application of this dataset of Alu insertion polymorphisms to a large sample size of wild populations could potentially resolve some of the complex issues influencing the current taxonomy. The Alu insertions reported in this study will provide researchers with an additional resource for the study of Aotus phylogenetic and population genetic relationships, bringing more insight into the elusive “night monkeys” to further facilitate conservation strategies.This study provides a comprehensive dataset of


Alternaria alternata was the predominant fungal aeroallergen among the studied patients (72.2%), which was followed by Aspergillus fumigatus (53.45%), Penicillium notatum (52.6%), Candida albicans (34.5%), and Aspergillus niger (25%). Conclusion: Mixed mold sensitization ranked fourth among the most frequent aeroallergens in airway-allergic patients, and Alternaria alternata was the most frequently encountered fungal aeroallergen in the Zagazig locality.Background: Airway allergies such as asthma and allergic rhinitis, as well as their comorbidities, are increasing worldwide, causing significant socioeconomic health burdens to societies. It is estimated that between 3% and 10% of the population is allergic to fungi. The type of fungal sensitization varies from one geographical region to another. The present study aimed to identify the common fungal aeroallergen sensitization patterns among airway-allergic patients residing in the Zagazig locality, Egypt, in order to obtain a better understanding of fungal allergy, in addition to improving the awareness and management strategies for those patients. Methods: The present cross-sectional study included 200 allergic rhinitis and asthma patients. Sensitization to fungal aeroallergens was evaluated by skin prick testing and in vitro measurement of total and specific immunoglobulin E. Results. As determined by a skin prick test, 58% of the patients studied were allergic to mixed molds.  Atopic diseases (immunoglobulin E (IgE)-mediated allergic diseases), such as allergic rhinitis, asthma, and atopic dermatitis, have been increasing in recent years. Collectively, they affect approximately 20% of the population worldwide . AllergeAspergillus, Alternaria, Cladosporium, Penicillium, Ulocladium, Fusarium, and yeast spores were the most frequently air-suspended fungal spores [2 limit proposed by the World Health Organization (WHO) [Cladosporium and Alternaria [According to past reports from Egypt and Saudi Arabia, l spores ,4,5. Morl spores . In Egypon (WHO) ,8. Tempeternaria . Penicillium, Aspergillus, Cladosporium, and Alternaria [Cladosporium and Alternaria have seasonal maxima [Although the precise prevalence of sensitization to fungi is unknown, it is estimated to range between 3% and 10% among the general population (depending extensively on the weather conditions of the area under study) . Accorditernaria . The spol maxima .Fungal spores are extensively air-suspended indoors and outdoors. Among the over 100,000 reported fungal species, a few hundred have been identified as opportunistic pathogens ,15. ExpoPoor asthma control is correlated with fungal sensitization, resulting in recurrent asthma attacks , an incrAspergillus niger (A. niger), Aspergillus fumigatus (A. fumigatus), Alternaria alternate , Penicillium notatum (P. notatum) and Candida albicans .Since the concentration of fungal spores in the atmosphere surpasses 1000 times pollen counts  and fungThe present cross-sectional study included airway allergy patients from attendants to the AIU, Faculty of Medicine, Zagazig University, Zagazig (30°33′59.99″ N and 31°29′59.99″ E), which is located 100 kilometers northeast of Cairo (30°03′17.40″ N and 31°13′26.40″ E) , the capAll patients were subjected to complete clinical history, including sociodemographic data. The diagnosis of allergic asthma and allergic rhinitis adhered to the Global Initiative of Asthma (GINA) and Allergic Rhinitis and its Impact on Asthma (ARIA) guidelines, respectively ,32. ExclAlternaria, Aspergillus, Penicillium, and Cladosporium species), mixed mites (Dermatophagoides pteronyssinus and Dermatophagoides farinae), date palm pollen, smoke, hay dust, wool, cotton, mixed feather  cat hair, and dog hair. Coca’s extracted aeroallergens were prepared and standardized under complete aseptic conditions at the AIU laboratory, as previously described ,34. PrepAccording to the SPT practical guide of the European Academy of Allergy and Clinical Immunology (EAACI), the SPT was performed, read, and interpreted . HistamiFor immunoassays, 3 mL of whole peripheral blood was collected. After centrifugation, the serum was stored at −20 °C until processing. Total IgE was measured using an indirect enzyme-linked immunosorbent assay according to the manufacturer’s instructions . Total IgE serum levels up to 150 IU/mL were considered normal.Dermatophagoides farinae, Dermatophagoides pteronyssinus, A. alternata, A. niger, A. fumigatus, P notatum, C. albicans, mixed grasses, birch, cat epithelium, dog epithelium, feather mixture, and cockroach) was analyzed by immunoblot assay according to the manufacturer company protocol . Serum-specific IgE was analyzed by a rapid scanner . The test was considered valid if the process control on each strip’s first position was colored. Specific IgE level ≥ 0.35 IU/mL was deemed positive.The serum aeroallergen-specific IgE for 13 common aeroallergens . Quantitative variables were displayed as mean ± standard deviation (SD) and median (min–max). Nominal and ordinal variables were displayed as frequency and percentage. Pearson Chi-Square (χ2) and Fisher’s exact tests were used when appropriate to analyze the association between dependent variables  and independent variables . The Mann–Whitney U test was used to compare the median difference between total IgE serum levels (dependent variable) in the dichotomous variable (independent variable). Spearman’s correlation coefficient (r) was used to determine the degree and direction of association between total IgE serum level (dependent variable) and age (independent variable). p = 0.038). The sociodemographic and clinical characteristics of the studied patients are illustrated in Two hundred patients with allergic airway diseases and attending AIU for skin testing of aeroallergen sensitization were selected by systematic random sampling to participate in the present study. More than two-thirds of the participants (73%) were adults, with males outnumbering females by 60.5%. Approximately half of the participants (58.0%) were rural population. Over half of the participants (60.5%) were diagnosed with allergic rhinitis, while 27% were diagnosed with combined allergic rhinitis and asthma, all significantly correlated with allergic rhinitis (p = 0.001), and 5.2% of mixed mold SPT-positive patients were polysensitized to more than five aeroallergens other than mixed molds  . It was ed molds .Alternaria alternata was the most prevalent fungal aeroallergen in mixed molds sensitized-allergic patients (72.2%), which was followed by A. fumigatus (53.45%), P. notatum (52.6%), C. albicans (34.5%), and A. niger (25%). The results of fungal aeroallergen sensitizations by immunoblot assay were significantly correlated with mixed mold sensitization by SPT (p < 0.001) (< 0.001) . Additio< 0.001) . The monC. albicans sensitization was observed in adults (p = 0.04), allergic rhinitis patients (p = 0.02), and patients with associated allergic conjunctivitis (p = 0.05), with statistical significance. A. alternata sensitization was significantly associated with sensitization to A. fumigatus (p < 0.001), P. notatum (p < 0.001), A. niger (p < 0.001), and C. albicans (p = 0.035)  .p = 0.7), as tested by SPT   . In addiThe pathogenesis of atopic diseases depends on aeroallergens, which have recently gained popularity in clinical research worldwide . AlthougConsistent with previous studies ,38, SPT In line with previous reports ,37,39, oExcept for one patient, all patients sensitized to mixed molds allergen extract in the study (99.1%) displayed sensitization to other allergens, indicating that monosensitization to fungi is unusual . The incAspergillus spp. and Penicillium spp. are among the predominant indoor molds [Alternaria spp. and Cladosporium spp. are typically outdoors. In Egypt, Alternaria, Aspergillus, Cladosporium, Penicillium, and yeasts are the most abundant airborne fungi indoors and outdoors in both urban and rural areas [Alternaria spp. is air-suspended at lower concentrations than other fungal aeroallergen spores, it demonstrated the highest sensitization rate in atopic patients in the current and previous studies [A. alternata sensitization ranges from 3.6 to 39.4%, our results revealed that the sensitization rate to A. alternata (43%) was the highest among the studied population, which was followed by indoor fungal spores, A. fumigatus (32%) and P. notatum (30.5%), and C. albicans (23%). Both outdoor and indoor fungi demonstrated nearly the same trend of sensitization rates when correlating to the sociodemographic and clinical characteristics of the studied patients. Since sensitization to outdoor fungal spores is more prevalent than indoor fungal spores [A. alternata sensitization was evident in 72.4% of the SPT fungal-sensitized patients, which was comparable to Mari et al. (66.1%) [Alternaria spp. air-suspended spores were recorded in June, July, and August [Airborne fungi are present in indoor and outdoor environments, sometimes occupying 70% of the total microflora of air . Aspergior molds , while A studies ,45. Comp studies , who estl spores , A. alte (66.1%) . Furtherd August , coincidSPT is the most reliable and sensitive tool to diagnose IgE-mediated allergic diseases and is considered the gold standard for allergy testing. It is a low-cost, minimally invasive procedure with immediate results that is reproducible when performed by professional allergists . ConsideA. alternata, A. fumigatus, and C. albicans were the most prevalent monosensitizers .Grouping the fungal allergen-sensitized patient by the pattern of specific IgE immunoblot reactivity elucidated demonstrated a prominent trend to combined sensitization to different fungal allergens in 45.5%. In contrast, monosensitization to only one fungal allergen was observed in 14.5% of the patients. Alternaria spp. are probably sensitized to one or more other allergenic fungal species, our result revealed that sensitization to A. alternata was rampant in 86.2% of A. niger, 73.4% of A. fumigatus, 77% of P. notatum, and 56.5% of C. albicans sensitized patients. Although cross-reactivity among fungi is not fully understood, multiple fungal sensitizations are due to the sensitivity to multiple antigens, whether cross-reactive or not [In agreement with Amado et al. , who repe or not . Moreovee or not .Cladosporium allergen in the mixed mold extract used in SPT and its absence in the immunoblot assay panel. Conversely, six patients displayed specific IgE immunoreactivity on immunoblots despite their negative SPT results. Further analysis could identify these six patients as C. albicans monosensitizers. However, this finding can be explained in a binary manner: the studied patients are either sensitized to C. albicans, which is not supported by the limitation of relevant SPT or sensitized to a fungal species cross-reactive with C. albicans [Only two patients lacked specific IgE immunoreactivity to any fungal allergens on an immunoblot when mixed mold-sensitized patients were categorized according to their SPT reactivity patterns. This finding may be attributed to the inclusion of albicans .C. albicans sensitization in adults, despite the inability to detect a correlation between sociodemographic and clinical characteristics and fungal sensitization patterns. However, the increased prevalence of C. albicans sensitization was previously reported by Ezeamuzie et al. [C. albicans can be found in many niches in the body, including the digestive tract and vagina, as a commensal organism and less commonly as an opportunistic pathogen, albeit its role in airway allergy is debated [C. albicans sensitization and allergic rhinitis alone or in combination with asthma. Improper administration of antibiotics may result in commensal Candida overgrowth above the tolerance threshold, and hence, sensitization proceeds [C. albicans sensitization to allergic conjunctivitis. Therefore, these data should encourage further investigations on the relationship between exposure to Candida infections and the development of airway-allergic disease [A significant finding of the current study was the increased prevalence of e et al. , Fernánde et al. , Asero ae et al. , and Mare et al. . C. albi debated ,53. The proceeds ,54. In c disease .Although approximately two-thirds of patients sensitized to molds were residents in rural areas, there was no association between the residence of the studied patients and fungal sensitization patterns, which aligns with the findings of previous studies conducted in India  and the As previously reported, up to 40% of allergic rhinitis patients have combined allergic rhinitis and asthma , albeit Our results revealed that the total IgE serum level did not correlate with sociodemographic or clinical characteristics of the studied patients nor differed between fungal sensitization patterns. Despite early studies ,63 consiIn order to gain a better understanding of the mechanisms and pathways by which fungi trigger allergic airway diseases and exacerbate asthma, additional research is recommended. It was reported that frequent exposures to fungal allergens induce T helper 1, 2, and 17 immune responses and chronic inflammation in airways . ConservEventually, identifying fungal aeroallergen sensitization patterns in airway-allergic patients is essential to avoid environmental trigger exposure and reduce allergy symptoms. It paves the way for patient education and tailoring personalized treatment with specific allergen immunotherapy . A. alternata has the highest prevalence rate among allergenic fungi in airway-allergic patients. Patients sensitized to one fungal species tend to be sensitized to one or more other species suggesting cross-reactivity between different allergenic fungi. Fungal sensitization patterns can be screened for SPT and confirmed by fungal-specific IgE immunoassay. Identifying fungal aeroallergen sensitization patterns in the study locality will improve airway allergic patients’ management. In light of the environmental data of airborne fungal spores, it is essential for individualized allergen avoidance and patient education, tailoring a better immunotherapeutic approach and encouraging government control efforts.Sensitization to fungi is the fourth prevalent aeroallergen implicated in airway-allergic diseases in the Zagazig locality. Monosensitization to fungal aeroallergen is remarkably infrequent, whereas sensitization to


Noteriades Cockerell, 1931 is recorded from Vietnam for the first time. A new species, Noteriadeshangkia Tran, Engel & Nguyen sp. nov. is described and figured based on a series of females collected from the provinces of the northern and central highlands of Vietnam. The genus is briefly discussed and a new subtribe is established, Noteriadina Engel, Tran & Nguyen subtrib. nov. of Megachilini. Lastly, an identification key and distribution map are provided for those species occurring in Southeast Asia.The little-known megachiline genus  Noteriades Cockerell is a seemingly relict genus, with comparatively few species occurring in both temperate and tropical regions of sub-Saharan Africa and southern Asia (Osmiini and among the Heriades-group of genera owing to its distinctly hoplitiform body habitus and size (Griswold (1985)Noteriades has recently been recovered as the sister group to Megachilini ; and N.spinosus Griswold & Gonzalez, 2011 which have been found in India, Thailand, and Myanmar. A further four species, all described from northern India (Osmiini (perhaps even as synonyms of other species), as evidenced by the tridentate mandibles and seeming absence of a mediolongitudinal carina on the clypeus , Hanoi, Vietnam and the Division of Entomology (SEMC), University of Kansas Natural History Museum, Lawrence, Kansas, USA (SEMC). Adult morphological and color characters were examined with a Nikon SMZ745 stereomicroscope, while images were photographed with a Nikon SMZ800N digital stereomicroscope, and with an ILCE-5000L/WAP2 digital camera attached to the stereomicroscope. Stacked focus images were prepared using with Helicon Focus 7. Finally, all files were processed with Adobe Photoshop CS6. The morphological terminology used in the description follows body length: measured from the base of the antennal torulus to metasomal apex , head length: measured from the medioapical margin of the clypeus to the upper margin of the vertex , head width: measured at the widest point of the head across the compound eyes , eye width: the greatest width of the compound eye (in profile), genal width: the greatest width of the gena (in profile), mesosomal width: measured between the outer rims of the tegulae .Specimens examined in the present study are deposited in the hymenopteran collections of the Institute of Ecology and Biological Resources  refer to numbered flagellomeres, metasomal sterna, and metasomal terga, respectively. The classification of Taxon classificationAnimaliaHymenopteraMegachilidaeEngel, Tran & Nguyensubtrib. nov.55D3BAE4-09B7-539F-81EA-0AB46D953077http://zoobank.org/AA0BBF9D-5715-4ADF-895C-0776BF3C9F24Noteriades Cockerell, 1931.N.pulchripes); propodeum wholly vertical, without basal subhorizontal zone; outer surfaces of pro- and mesotibiae apically with an acute angle and distinct notch anteriorly, therefore appearing bispinose in apical view; arolia present on all legs in both sexes (absent in Megachilina except Matangapis Baker & Engel and Heriadopsis Cockerell). Metasomal tergum I carinate dorsally at angle between anterior- and dorsal-facing surfaces; tergum VI of female nearly vertical except for apical flange-like hyaline margin, without preapical carina, tergum VI of male without preapical carina (present in Megachilina); terga V and VI of male strongly curved ventrally , covering tergum VII and sterna III–VI (no so in Megachilina); sternum I of male produced over its apical margin subapically, forming double carina (not so in Megachilina); volsella distinct, with well-developed digitus and cuspis, with heavily sclerotized denticles resembling those of short-tongued bee families and Pararhophites Friese. Refer to Megachilini.Small to modest-sized (4.5–10.2 mm), non-metallic, hoplitiform bees lacking integumental maculation; mandible of female quadridentate, without differentiated cutting edges, mandible of male bidentate; malar space linear; clypeus slightly projecting over clypeal-labral articulation; clypeus and often supraclypeal area with mediolongitudinal carina; paraocular area with dense appressed pubescence; preoccipital carina complete. Pronotum not enlarged nor surrounding mesoscutum anteriorly; pronotal lobe and omaulus carinate, with defined omaular surface; mesoscutellum flat, carinate posteriorly, overhanging metanotum Megachiletricarinata Bingham, 1903, by original designation. Cockerell, 1931: 332. Type species: vide supra).As for the subtribe ; the rim of the antennal torulus mesodorsally extended into a short lamellate tubercle ; mesosoma approximately as long as broad (mesosoma longer than broad in N.jenniferae). In addition, the new species differs from both N.jenniferae and N.spinosus by the generally shiny face and mesoscutum, which is matte in the latter two species.The female of this species is most similar to that of ♀: Body length 8.0–8.5 mm (holotype = 8.5 mm), forewing length 5.5–6.0 mm (holotype = 6.0 mm).Structure. Head slightly broader than long, approximately 1.1× as broad as long (Fig. ong Fig. . Compounong Fig. . Supraclong Fig. , scape aong Fig. ; mesoscuong Fig. . Forewinong Fig. , margin ong Fig.  and dorsSculpturing and texture. Integument of head and mesosoma generally shiny. Mandible and labrum irregularly punctate, punctures slightly coarser on mandible basally, outer ridges smooth and shiny. Clypeus with contiguous punctures of unequal sizes, puncture sizes laterally and along base larger than on remainder of surface (Fig. ace Fig. . Supraclace Fig. . Puncturace Fig. . Mesepisace Fig. . AnterioColor. Body black except antenna beneath, tegula, tarsi, and metasomal sterna apical margins dark reddish brown. Wings light brown with faint green mixed coppery highlights in ventral view, membrane of marginal cell and apex darker brown than remainder of remigium and lighter in radial and first cubital cells; veins brown to dark brown, prestigma and pterostigma dark brown.Pubescence. Paraocular area from epistomal sulcus to slightly above antennal toruli with long, dense, plumose, appressed, white setae, some setae tinged yellowish (Fig. ish Fig. . Apical ish Fig. , otherwiish Fig. .Latet.♂: The specific epithet is a toponym for the locality at which the holotype was collected, the Hang Kia commune in Hoa Binh Province. The name is treated as a noun in apposition.N.hangkia in the northern and central highlands of Vietnam extends the distribution of the genus Noteriades in Southeast Asia (Fig. The discovery of sia Fig. . In factCharacters for the key were extracted from the original descriptions of the species .


Pseudomonas aeruginosa are type II TA systems. Type II TA systems have various physiological functions, such as virulence and biofilm formation, protection host against antibiotics, persistence, plasmid maintenance, and prophage production. Here, we review the type II TA systems of P. aeruginosa, focusing on their biological functions and regulatory mechanisms, providing potential applications for the novel drug design.Toxin–antitoxin (TA) systems are typically composed of a stable toxin and a labile antitoxin; the latter counteracts the toxicity of the former under suitable conditions. TA systems are classified into eight types based on the nature and molecular modes of action of the antitoxin component so far. The 10 pairs of TA systems discovered and experimentally characterised in  Toxin–antitoxin (TA) systems were first discovered on a conjugative plasmid in 1983, functioning as plasmid maintenance systems by post-segregational killing (PSK) . In typeThe toxin inhibits an essential cellular process (kills cells or inhibits their growth), while the antitoxin counteracts the toxicity of its cognate toxin ,13. TA sPseudomonas aeruginosa is one of the ESKAPE pathogens  and is a critical priority pathogen [P. aeruginosa from the NCBI Genome database, most of which were in chromosome sequences (94%) and belonged to type II TA systems  [P. aeruginosa, 8 of which are located on the chromosome [P. aeruginosa.pathogen . Dai et 08 hits) . To dateromosome ,24,25,26romosome ,28. ThesP. aeruginosa  overlapping the −35 region of the pfiTA promoter, thereby repressing the expression of the pfiTA operon [relBE operon is regulated by both the antitoxin RelB and the TA complex RelBE; antitoxin RelB is a weak repressor, whereas RelE is an efficient co-repressor to further increase transcriptional regulation [pumAB, pacTA, parDE, and hicAB TA systems in P. aeruginosa are unclear. The stoichiometries of TA complexes depend on the toxin/antitoxin ratio and affect the affinity for the operon. In the presence of excess antitoxin relative to a toxin, the operon is weakly repressed. However, excess toxin leads to the formation of saturated TA complexes, which cannot bind operons [Ten pairs of type II TA systems have been experimentally verified in ruginosa , as follruginosa . In the al level . Antitoxal level . The antal level . The higA operon . Interesgulation ,31. Howe operons ,32,33.P. aeruginosa, most type II TA systems are encoded on the chromosome [In romosome ,24,25,26romosome ,28. Thesromosome ,35,36, promosome , persistromosome , plasmidromosome , and proromosome   genes [mvfR, exsA, and amrZ by directly binding to their promoter regions, and it controls pyocyanin synthesis, T3SS, and type VI secretion system (T6SS) expression [mvfR is a key virulence-related regulator of P. aeruginosa that activates the expression of the phnAB and pqsA-E operons [exsA is the master regulator that activates the expression of all the T3SS genes [amrZ is a global T6SS transcriptional regulator that activates H1-T6SS and H3-T6SS but represses H2-T6SS expression by binding to the promoter regions of T6SS genes [HigBA is prevalent in  species ,22. The  species   genes . HigB moS) genes . The antpression ,44. mvfR operons . exsA isSS genes . amrZ isSS genes .Escherichia coli K-12 [relBE genes were detected in 100% of P. aeruginosa [relBE expression is increased in isolates that are sensitive, compared with isolates that are resistant to aztreonam [relBE promotes persister cell formation in biofilms in the presence of ciprofloxacin and colistin in P. aeruginosa [The RelBE TA system was discovered in the chromosome of oli K-12 ,49, and ruginosa . RelBE Truginosa . relBE eztreonam . Zadeh eruginosa . RelBE Truginosa .P. aeruginosa isolate 39016, 3504 bp upstream of prophage att sites [prrA significantly increased biofilm formation and reduced motility [amrZ transcript level was significantly decreased in a ΔprrA mutant, a global regulator of genes associated with a virulence that indirectly modulates the c-di-GMP level by repressing diguanylate cyclase genes or activating phosphodiesterase genes [prrTA system is involved in prophage regulation and production. Most prophage genes are significantly downregulated in the ΔprrA mutant, and phage production and infectivity are significantly lower than those in the wild type [The PrrTA TA system is encoded by the chromosome of the clinical tt sites . Deletiomotility . The amrse genes ,54. Morese genes . In addiild type .P. aeruginosa PA14. The acetyltransferase toxin PacT inhibits the DNA-binding activity of Fur (ferric uptake regulator) to maintain iron homeostasis by directly binding to the HTH domain of Fur [P. aeruginosa. Compared with the wild-type, pyocyanin production and biofilm formation were markedly decreased in the ΔpacT and ΔpacTA mutants [Yersinia pestis [Stenotrophomonas maltophilia [P. aeruginosa; compared with the wild type, the ΔpacT and ΔpacTA mutants caused lower mortality.The novel type II TA system PacTA has been characterised by n of Fur . Fur is n of Fur . Moreove mutants . Fur repa pestis  and Stentophilia ; PacTA mE. coli and Salmonella species [P. aeruginosa, the toxin ParE promoted survival in the presence of quinolone antibiotics, e.g., ciprofloxacin, levofloxacin, and novobiocin [The ParDE TA system was characterised in plasmid RK2 as a plasmid stabilisation element . This TA species . ParDE c species . In P. avobiocin . Howevervobiocin . ParDE ivobiocin . This isvobiocin . This phP. aeruginosa strain PA14. The toxin ResA belongs to the RES family, and the antitoxin XreA is a xenobiotic response element (Xre) [+ level, a key component of the respiratory machinery. Therefore, a reduced intracellular NAD+ level could increase antibiotic resistance. Overproduction of NAD+ counteracts the effect of overexpression of ResA on persister formation, suggesting that ResA/XreA contributes to persister formation by reducing the intracellular NAD+ level [resA did not affect persister formation due to additional TA systems or other determinants involved in persister formation.Persister cells are slow-growing or growth-arrested cells that can resume growth after the stress disappears . Persistnt (Xre) . ResA/Xrnt (Xre) . OverexpD+ level . HoweverccdAB and hok-sok, stabilise plasmids by PSK [P. aeruginosa are unclear. The PumAB TA system is encoded from the conjugative plasmid pUM505 of P. aeruginosa strain PUM503 [merRTPFADE), chromate resistance operon (chrBAC), and ciprofloxacin resistance gene (crpP) [pumAB-carrying plasmids decreased in number after 216 generations and were undetected after 432 generations; by contrast, pumAB-carrying plasmids were not significantly affected [pumAB can confer post-segregational plasmid stability via PSK [P. aeruginosa. Additionally, the PumA toxin confers P. aeruginosa virulence and increased Caenorhabditis elegans and mouse mortality rates [Plasmid maintenance is a function of plasmid-encoded type II TA systems . The twos by PSK ,64. PSK s by PSK . The funn PUM503 . pUM505 e (crpP) . Non-pumaffected . Therefo via PSK . No mechty rates .E. coli [Shewanella oneidensis [P. aeruginosa PAO1 strain [pfiT increased Pf4 phage production by approximately 100,000-fold compared with the wild type. In the ∆pfiT mutant, the phage replication initiation gene PA0727 was induced but not the phage repressor gene pf4r. Therefore, PfiT regulates Pf4 phage production by inducing the expression of the replication initiation gene PA0727 [PfiT is involved in phage immunity and coordinates the phage repressor Pf4r in conferring immunity to Pf4 [P. aeruginosa WK172 is involved in fighting against phage infection. In excess, the prophage Cro-like antitoxin CrlA inhibited infection by lytic Pseudomonas phages  because CrlA inhibited phage replication by binding to crlTA palindrome-like sequences in the phage genome [Prophages and satellite prophages are widely distributed among bacteria, in which they confer various phenotypic traits to their hosts, including pathogenicity , antibioE. coli  and Sheweidensis . TA syst1 strain . The Pf41 strain ,72. Delee PA0727 . In addiy to Pf4 . The chre genome . The CrlBacterial TA systems have considerable biotechnological potential ,73,74. FTA systems can be used as positive selectable markers for ensuring the stability of mobile genetic materials . An examP. aeruginosa, the biological functions of TA systems have been focused on recently. Ten pairs of type II TA systems have been identified in P. aeruginosa. In P. aeruginosa, type II TA systems enhance fitness and have multiple biological functions. For instance, the HigBA TA system is associated with virulence and biofilm formation, persister formation, T3SS, and T6SS. Interestingly, the ParDE TA system confers protective or toxic effects depending on the toxin concentration, but the molecular basis remains unknown for the switch from protective effects at low concentrations to toxic effects at higher concentrations. The HicAB TA system has known functions in other bacteria, e.g., virulence, persister cell formation, and stress responses [P. aeruginosa [Although TA systems have been widely distributed and well studied in pathogenic bacteria for decades, in esponses ,81; howeruginosa .P. aeruginosa, the antitoxin HigA regulates the transcriptional regulatory genes such as mvfR [exsA [amrZ [The antitoxin can autoregulate TA operon transcription and also regulate other key regulatory genes. The antitoxin is a negative regulator and participates in repressing the expression of other key regulatory genes ,83,84. I as mvfR , exsA [3fR [exsA , and amrsA [amrZ , which a


Certain patients at low risk of clinically significant esophageal varices by Baveno criteria can safely avoid screening esophagogastroduodenoscopy (EGD) , 2. HoweThe COVID-19 pandemic has raised challenges for procedure units resulting in EGD delays and case backlogs, which has resulted in a consensus statement urging providers to consider earlier initiation of pharmacologic variceal prophylaxis instead of EGD in appropriate populations , 6. WhilWe generated a survey cohort using a data repository at our tertiary center. Our inclusion criteria included adult patients (≥18 years) recently seen in our hepatology clinic for cirrhosis care with a platelet count of <150,000 and a valid email address for recruitment. Manual chart review of respondents was performed to confirm inclusion criteria and to collect demographic data.We utilized Research Electronic Data Capture (RedCap) for survey creation and email distribution. A total of 944 surveys were sent, with a response rate of 21.5% (203 patients). The survey began with a summary of our current screening paradigm and standard of care for esophageal varices, and of ongoing discussions that the pre-primary use of BB (in lieu of EGD first for refinement of use) may be appropriate in select patients. Survey questions utilized Likert-scale scoring to investigate patient preferences surrounding BB, EGD, and concerns related to COVID-19  and 2.Our primary outcome was patient preference to trial BB first over EGD screening, which was asked as “I would like to try beta blocker first and then decide.” Multinomial logistic regression was also performed to assess which factors were associated with willingness to trial BB first vs EGD first based on patient-reported preferences. Data are presented as odds ratios, utilizing neutral responses as the reference group. Covariates examined included sex, Model of End-stage Liver Disease-Sodium (MELD-Na) score, exposure histories , as well as general perceptions surrounding EGD, BB, variceal bleeding, and COVID-19.Our surveyed cohort demographics  includedSurvey results showed that 20.2% of patients preferred BB first, 45.8% were neutral, and 34.0% favored EGD first. While only 12.5% felt that avoiding EGD was important , 50.2% felt that avoiding another pill was important , 44.8% were concerned about BB side effects , and 37.9% stated that they preferred avoiding the hospital due to COVID-19 . In We analysed the impact of our covariates and Likert-scale questions surveyed on willingness to trial pre-primary BB by multinomial logistic regression . A prefePre-primary BB use for select patients at low risk of varices requiring treatment is attractive in the context of emerging data suggesting the ability to prevent not only variceal bleeding but also overall hepatic decompensation , 4. The Our survey indicates that the majority of our cohort is either neutral or agreeable to pre-primary BB. Factors decreasing BB acceptance include the preference of avoiding medications and concern for BB side effects. Notably, COVID-19 did not have a statistically significant impact on preferences, suggesting that shared decision-making may be less effective if centered on COVID concerns. Above all, the desire to receive personalized physician recommendations, even if this differs from current guidelines, persuades many patients. This emphasizes how critical counseling and shared decision-making are in the context of significant neutrality.Our data identify key patient characteristics that influence preference for BB use but must be interpreted within the context of the study design. The response rate was limited by certain survey factors. We note that some participants did not ultimately complete the survey, which may have been due to the moderately lengthy educational component, as well as the number of questions. The lower response rate may affect the overall generalizability and reliability of statistical significance. Many patients also had prior exposure to EGD and it is thus unclear how these findings generalize to the rare persons without EGD experience. Future directions include survey recruitment of patients with a new or recent diagnosis of cirrhosis without prior exposures to BB or EGD to remove those influences, including a possible prospective study design.Most patients we surveyed are neutral to pre-primary BB for variceal bleed prophylaxis, which is attractive during the current pandemic and beyond. Our data indicate that patients who are highly concerned about pill burden and BB side effects may be less suitable for a BB-first strategy, though all patients would benefit from a direct explanation of the rationale by their trusted provider.Gastroenterology Report online.Z.C. and E.B.T. conceptualized and designed the study. Z.C. performed study implementation. Z.C. and I.K. participated in data acquisition. Z.C., J.L., and E.B.T. performed data analysis and interpretation. Z.C. drafted the manuscript. All authors read and approved the final manuscript.E.B.T. is supported by NIDDK K23 DK117055.E.B.T. has served on advisory boards for Mallinckrodt, Kaleido, Rebiotix, Novo Nordisk, and Bausch Health; consulted for Allergan, Novartis; and has received unrestricted research grants from Valeant, Gilead. All other authors have none declared.goac015_Supplementary_DataClick here for additional data file.


Lactiplantibacillus  plantarum. Antimicrobial resistance (AMR) genes were bioinformatically analyzed using all Lp. plantarum genomes available in GenBank, which showed that AMR genes are present differently depending on Lp. plantarum strains. Bioinformatic analysis demonstrated that some mobile genetic elements such as prophages and insertion sequences were identified in the genome of strain Q180, but because they did not contain harmful genes such as AMR genes and virulence factor (VF)- and toxin-related genes, it was suggested that there is no transferability of harmful genes. The minimum inhibition concentrations of seven tested antibiotics suggested by the European Food Safety Authority guidelines were slightly lower than or equal to the microbiological cut-off values for Lp. plantarum. Strain Q180 did not show hemolytic and gelatinase activities and biogenic amine-producing ability. Taken together, this study demonstrated the safety of strain Q180 in terms of absence of AMR genes and VF- and toxin-related genes as a probiotic strain.The safety of the probiotic strain Q180, which exerts postprandial lipid-lowering effects, was bioinformatically and phenotypically evaluated. The genome of strain Q180 was completely sequenced, and single circular chromosome of 3,197,263 bp without any plasmid was generated. Phylogenetic and related analyses using16S rRNA gene and whole-genome sequences revealed that strain Q180 is a member of  Probiotics are defined as “live microorganisms which when administered in adequate amounts confer a health benefit on the host” FAO/WHO 2002). While probiotics have several known benefits, there have been safety issues related to their use in humans and animals. In 2002, the Food and Agriculture Organization/World Health Organization (FAO/WHO) reported that probiotics may cause side-effect such as systemic infection, deleterious metabolism, and excessive immune response in susceptible subjects or exhibit harmful gene transfer (FAO/WHO 2002). Furthermore, the European Food Safety Authority (EFSA) announced safety assessment guidance for probiotics by focusing on antimicrobial resistance (AMR)  and made002. WhilLactobacillus plantarum, one of the lactic acid bacteria (LAB) widely used worldwide as a probiotic [Lactobacillus species on the basis of whole genome sequencing data and newly denominated as Lactiplantibacillus (Lp.) plantarum [Lp. plantarum using molecular biological approaches, microbiological tools, and bioinformatics analyses [Lp. plantarum strains cannot be guaranteed without verification, as some Lactobacillus strains have been reported to exhibit AMR and may not meet the EFSA guideline criteria. The most representative case was the intrinsic resistance of Lp. plantarum to vancomycin [Lp. plantarum strains [robiotic -7, was dlantarum . Many reanalyses -13. Howencomycin -16, causncomycin . Intrins strains -20. The In our previous studies, strain Q180 isolated from the feces of a healthy Korean adult was found to exhibit blood triglyceride lowering effects in vivo and in clinical trials -23. We iLp. plantarum ATCC 14917T, Enterococcus faecium ATCC 19434T, and Staphylococcus aureus ATCC 6538 were purchased from their culture collection centers and used as reference strains for the comparison of phenotypic properties. Strains Q180, ATCC 14917T, and ATCC 19434T were cultured in deMan-Rogosa-Sharpe  broth, and strain ATCC 6538 was cultured in tryptic soy broth (BD) at 37°C. The bacterial cultures were mixed with 20% skim milk (BD) at a 1:1 ratio and preserved at −80°C. All test experiments were performed using freshly cultured cells from frozen stocks and not from consecutively subcultured cells.Strain Q180 was provided by the Korea Food Research Institute  , and Lp.de novo assembled using the Hierarchical Genome Assembly Process software (ver. 3.0) and the PBcR pipeline of Celera Assembler [The genomic DNA of strain Q180 was extracted and completely sequenced using the combination of PacBio RS II and Illumina HiSeq 2500 platforms at Macrogen (Korea), as previously described . In briessembler .http://www.ezbiocloud.net/). For the genome-based phylogenetic analysis, 92 housekeeping core genes from the genomes of strain Q180 and closely related type strains were extracted using the UBCG pipeline (www.ezbiocloud.net/tools/ubcg) [Taxonomic identification of strain Q180 was conducted through phylogenetic analyses based on the 16S rRNA gene and whole-genome sequences. For the 16S rRNA gene sequence-based phylogenetic analysis, the 16S rRNA gene sequences of strain Q180 and closely related type strains were aligned using the fast secondary-structure-aware infernal aligner available in the ribosomal database project . A maximls/ubcg) , and an www.ezbiocloud.net/sw/oat) [http://ggdc.dsmz.de/distcalc2.php) [The genome relatedness between strain Q180 and closely related type strains was evaluated through average nucleotide identity (ANI) and digital DNA-DNA hybridization (DDH) analyses using the Orthologous Average Nucleotide Identity Tool software (/sw/oat)  and the lc2.php) , respectLp. plantarum DSM 20174T were functionally classified into clusters of orthologous groups (COG) categories using eggNOG-mapper (ver. 5.0)[The circular map of the assembled genome of strain Q180 was visualized using a web-based CGview program . The comver. 5.0).Lp. plantarum strains, including strain Q180 (CP073753) and Lp. plantarum DSM 20174T (CP039121), available in GenBank were analyzed. All genomes of Lp. plantarum were downloaded from the GenBank database, and AMR genes in each genome were searched through BlastX analysis using Diamond software (ver. 0.9.26.127) [AMR genes present in the genomes of all .26.127)  based on.26.127) , Antibio.26.127) , and ResQ180 CP0753 and LpLp. plantarum DSM 20174T, E. faecium ATCC 19434T (UFYJ00000000), and S. aureus ATCC 6538 (CP020020–21) based on the virulence factor database (VFDB) using BlastX available in the Diamond software [VF- and toxin-related genes, including those associated with enterotoxin, leucotoxin, cytolysin, cytotoxin K, hemolysis, BA production, hyaluronidase, aggregation, enterococcal surface protein, endocarditis antigen, collagen adhesion, cereulide, sex pheromone, and serine protease were searched in the genomes of strain Q180, To assess the transferability of AMR/VF genes in strain Q180, prophages and insertion sequences (including transposons), which are representative MGEs, were investigated using PHAge Search Tool Enhanced Release (PHASTER)  and ISfi2 and mitomycin C was added to be a final concentration of 0.6 μg/ml, followed by additional incubation at 30°C for 18 h. Phage DNA was isolated from the cell culture supernatants, according to the phage DNA extraction protocol described previously [The inducibility of the putative prophage sequences as active phages in strain Q180 was evaluated using a mitomycin C approach, as described previously . In brieeviously , and detLp. plantarum ATCC 14917T for seven antibiotics  was assessed by measuring the MIC of each antibiotic using the E-test strip and broth microdilution susceptibility assay methods. The MIC tests were performed in triplicates for each antibiotic. For the MIC assay using E-test strip, bacterial suspensions were spread onto agar media containing LAB susceptibility medium  supplemented with 1.5% agar (BD). E-test strips  were applied to the center of each agar plate, which was incubated for 24 h at 37°C. The lowest concentrations of antibiotics that inhibited the visible growth of strain Q180 and Lp. plantarum ATCC 14917T on LSM agar were determined as MICs. The MICs of each antibiotic for strain Q180 and Lp. plantarum ATCC 14917T were also assessed according to the standard broth microdilution susceptibility assay protocol recommended by the National Committee for Clinical Laboratory Standards [Phenotypic resistance of strain Q180 and tandards .Lp. plantarum ATCC 14917T, E. faecium ATCC 19434T, and S. aureus ATCC 6538 were used as negative or positive controls. Frozen stocks of bacterial strains were streaked on MRS agar and incubated at 37°C for 24 h. Single colonies grown on MRS agar were inoculated into MRS broth, and an aliquot of the cultured broth was used for the tests. All experiments were performed in triplicates.Phenotypic tests for hemolytic and gelatinase activities of strain Q180 were performed, and BA-producing ability of strain Q180 was investigated. α, β, and γ) of the strains was evaluated based on the formation of hemolysis zones around colonies grown on blood agar [To evaluate hemolytic activity, each strain was streaked on blood agar containing 5% (w/v) sheep blood  and the plates were incubated at 37°C for 48 h. The hemolytic activity (ood agar  To test 18 column (250 × 4.6 mm) and a fluorescence detector (RF-10AXL) as described by Brückner et al. [The BA-producing ability of each strain was assessed according to a previously described procedure . In brier et al. .Lp. plantarum DSM 20174T and Lp. argentoratensis DSM 16365T within the genus Lactiplantibacillus  with Lp. argentoratensis DSM 16365T than the thresholds, suggesting that they are distinct from Lp. argentoratensis at the species level but exhibit 100% similarity in 16S rRNA gene sequences.The genome of strain Q180 was completely sequenced with high quality and taxonomically identified through 16S rRNA gene- and whole genome-based phylogenetic analyses. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain Q180 formed a phylogenic lineage with both bacillus , and shoM 16365T , suggestM 20174T , which wLp. plantarum (DSM 20174T) , COG categories, and G+C contents, are graphically depicted in  20174T) . The gen similar .Lactobacillus strains harbor AMR genes that are involved in mediating resistance to several antibiotics, including β-lactam, macrolide, aminoglycoside, chloramphenicol, and tetracycline [48tetW, tetM, and tetO have been frequently identified in Lactobacillus species [Lp. plantarum strains that are widely used as probiotics were often deemed as safe. However, several studies have reported that safety issues such as AMR, virulence, and toxin-producing ability vary depending on the strain even within the same species [Lp. plantarum strains, AMR genes in all Lp. plantarum genomes available in GenBank were bioinformatically analyzed based on the CARD, ARG-ANNOT, and ResFinder databases , which suggests that strain Q180 and Lp. plantarum DSM 20174T are considered safe as probiotics in the AMR gene issues.During the evaluation of the safety of probiotic strains, their phenotypic properties related to AMR are considered important (WHO 2019). As the genomic analysis of microbes provides information on AMR genes as well as their potential transferability, genomic analysis is necessary for the evaluation of the potential risks of new probiotic strains. Previous genomic analysis studies have shown that many acycline , 48-50.  species . In the  species , 53, sugatabases . BioinfoLp. plantarum DSM 20174T, E. faecium ATCC 19434T, and S. aureus ATCC 6538  and three incomplete phage sequences were identified in the genome of strain Q180 , suggestLp. plantarum were identified in the genome of strain Q180 (Lp. plantarum DSM 20174T), it may have low transferability of AMR/VF or toxin genes via MGEs.The bioinformatic analysis also showed that two types of insertion sequences (IS1182 and ISL3) annotated as transposases probably originating from ain Q180 . As no ALactiplantibacillus, classified as formerly Lactobacillus [Lactiplantibacillus strains have been shown to exhibit AMR [Lp. plantarum strains commonly exhibit intrinsic resistance to vancomycin [Lp. plantarum strains to vancomycin and streptomycin, non-intrinsic resistance to other antibiotics has been reported in some Lp. plantarum strains [Lp. plantarum genomes also showed that some Lp. plantarum strains harbor AMR genes, which are different depending on strains , similar to Lp. plantarum DSM 20174T and E. faecium ATCC 19434T, while S. aureus ATCC 6538 exhibited clear hemolytic activity (β-hemolysis)  . This obmolysis) .Lp. plantarum DSM 20174T and E. faecium ATCC 19434T, while S. aureus ATCC 6538 showed clear gelatinase activity (Lp. plantarum strains showed no gelatinase activity [Gelatinase is a hydrophobic metalloprotease that cleaves insulin, casein, hemoglobin, collagen, and gelatin. Because it has been reported that pathogenic bacteria such as enterococci responsible for endocarditis and bacteremia have the ability to penetrate the tissue through their gelatinase activities , gelatinactivity . This reactivity . Previouactivity , 60.Lp. plantarum DSM 20174T and S. aureus ATCC 6538 and that E. faecium ATCC 19434T had the ability to produce tyramine (Lp. plantarum strains do not have the ability to produce BAs [BAs such as histamine, tyramine, cadaverine, and putrescine, mainly formed by the decarboxylation of their corresponding amino acids, act as neurotransmitters in organisms; however, excessive intake of BAs can cause diseases or disorders such as diarrhea, vomiting, sweating, and tachycardia . Here, wtyramine . This obtyramine . Recent duce BAs -64. TakeLp. plantarum that has been widely used as probiotics. Second, the whole-genome of strain Q180 was bioinformatically analyzed, and strain Q180 did not harbor potentially harmful genes such as AMR genes and VF- and toxin-related genes. Third, phenotypic tests for the presence of some virulence properties were performed, and strain Q180 was found to exhibit no AMR, hemolytic and gelatinase activities, and BA-producing ability. In conclusion, strain Q180 may be safe for human health and can be used as a potential probiotic strain to ameliorate high blood triglyceride levels. Although bioinformatic and phenotypic analyses conducted in this study can provide the basic safety information for probiotic strains, further systematic and acceptable approaches that evaluate the safety of probiotic strains should be conducted to confirm their safe and effective applications.In this study, the safety of strain Q180 known to exhibit postprandial lipid-lowering effect was evaluated through bioinformatic and phenotypic analyses to test its potential as a probiotic strain. First, strain Q180 was subjected to phylogenetic and genome-related analyses, which showed that it belongs to http://jmb.or.kr.Supplementary data for this paper are available on-line only at