E. coli from three major one health components  in Bangladesh. Samples were randomly collected from broiler chickens, broiler farm environments and hospitalized human patients from the same geographical area. Isolation and identification of E. coli were performed following standard bacteriological techniques. Antimicrobial susceptibility testing (AST) was performed by disk diffusion and broth microdilution methods. Mutation at the quinolone-resistance determining region (QRDR) was analyzed by sequencing. Of 450 samples, a total of 287  E. coli strains was isolated, where 240  strains were phenotypically resistant to ciprofloxacin. The prevalence of ciprofloxacin-resistant E. coli in broiler chicken, broiler farm environments and hospitalized human patients are 77.6%, 88.8% and 89% respectively. In AST against nine antimicrobials, all the isolates were found to be multidrug-resistant (MDR). The minimum inhibitory concentration (MIC) of ciprofloxacin was ranged from 4 to >128mg/L. Point mutations were detected in several sites of QRDR, specifically at 83 and 87 amino acid positions in gyrA gene, and 56, 57, 78, 80 and 84 amino acid positions in parC gene. Mutations resulted in amino acid substitutions. Phylogenetic analysis of gyrA and parC gene sequences showed a close relationship between the strains isolated from different sources. This study demonstrates a high prevalence of ciprofloxacin resistance in commensal E. coli in humans, animals and environment interface and their genealogically similarity poses an alarming public health consequence.The emergence of antimicrobial resistance in commensal bacteria poses a serious public health burden worldwide. Commensals can disseminate the resistance genes to pathogenic bacteria causing life-threatening infections. This cross-sectional study was designed to investigate the antimicrobial resistance pattern and molecular mechanism(s) of ciprofloxacin resistance in commensal  E. coli is one of the versatile commensals in warm-blooded animals’ intestinal tract. Commensal E. coli has shown resistance against numerous antimicrobials due to their widespread use in animals  by the Ethics Committee of Chattogram Veterinary and Animal Sciences University (CVASU), Bangladesh.A total of 450 samples were collected from the study population. Among them, 150 cloacal swabs from broiler chickens, 150 samples of farm environments and 150 rectal swabs from hospitalized humans. Samples from different environmental matrices are described in E. coli  followed by farm environments and hospitalized humans. as observed for broilers, farm environment  and hospitalized humans .A total of 287  59.2–68.1%) samples were tested positive for the presence of E. coli . The higE. coli (98.3%) and ciprofloxacin-resistant E. coli (E. coli (76.7%) and ciprofloxacin E. coli (73.3%) among other matrices (E. coli (54.1%) and ciprofloxacin-resistant E. coli (47.4%) were high in the farm environments of those farms reused litter materials. In hospitalized humans, the highest prevalence of E. coli (71.5%) and ciprofloxacin E. coli (67.3%) were isolated from the age group of more than 55 years. Male patients had the highest prevalence of E. coli (53.9%) and ciprofloxacin-resistant E. coli (46.1%). Simultaneously, the prevalence of E. coli (50.6%) and ciprofloxacin-resistant E. coli (44.3%) were high in non-diarrhoeic patients. A total of 48.6% samples from hospitalized humans those used ciprofloxacin for medication, were positive for ciprofloxacin resistant E. coli , followed by sulfomethoxazole (97.9%), and tetracycline (97.9%) (E. coli strains isolated from the human patients show the highest resistance against ampicillin (100%), enrofloxacin (100%) and sulfamethoxazole (100%)  . E. colie (100%) . In conte (100%) . On the E. coli isolates were detected in hospitalized humans (89%)  isolates were resistant to ciprofloxacin. The highest proportion of ciprofloxacin resistant ns (89%) , where tectively .E. coli strains from different sources is illustrated in heatmaps (All the isolates were found to be multidrug-resistant (MDR) as they showed resistance to more than two antimicrobials tested . Most isheatmaps . The denE. coli strains from human patients showed very high MIC (≥128mg/L)  was tested. Overall, 32.5% of the tested isolates show extremely high MIC values (≥128mg/L) against ciprofloxacin . Alarmin128mg/L) . The E.  samples .gyrA and parC genes. All the isolates were positive for gyrA and parC genes. As mutation(s) in the gyrA and parC genes, specifically in QRDR results in the development of resistance to the ciprofloxacin, we detected point mutation(s) in these two genes. Twenty isolates were selected randomly, covering all types of sample sources for sequencing the gyrA and parC genes. The gene sequences were submitted to the NCBI nucleotide sequence database, which are available under the accession number ON463777-96 for the gyrA gene and ON479036-65 for parC gene.A total of 240 phenotypically ciprofloxacin-resistant isolates were selected for PCR assays to detect gyrA gene  was substituted by leucine (TTG) due to a single point mutation in the amino acid codon. However, at 87 position aspartic acid (GAC) was replaced by either histidine (CAC) or asparagine (AAC) or glycine (GGC) or tyrosine (TAC).We detected point mutations at several sites of QRDR, specifically 83 and 87 amino acid positions in yrA gene . At 83 pparC genes, mutations were detected in 56, 57, 78, 80 and 84 amino acid positions  was swapped into cysteine (TGC).A Common similarity between isolates from three types of sources was the substitution of serine (TCG) by leucine (TTG) with a single point mutation in the 83 amino acid codon of gyrA gene sequences revealed that all the sequences were grouped into three monophyletic clades (parC gene sequences clustered into four monophyletic clades (Maximum likelihood (ML) phylogenetic tree of twenty c clades . Each ofc clades , where tE. coli isolated from hospitalized humans and animal sources. This study primarily focuses on the emergence of ciprofloxacin resistance in commensal E. coli followed by characterization of the mechanism of ciprofloxacin resistance phenomenon by mutation analysis in QRDR.AMR is an alarming public health burden worldwide, and it is concerning when commensal bacteria develop resistance to vital antimicrobial agents. Our study generated evidence of a high prevalence of AMR in commensal E. coli from broiler chicken (89.3%) and farm environment (53.3%) was recorded, which can be influenced by factors such as age, strains and husbandry system of broiler chicken. For instance, the prevalence of E. coli is higher in free-range chickens than caged chickens .  and green color represents the sensitivity profile. All the intermediately susceptible isolates were considered as susceptible. The dendrogram in the left side clustered the isolates based on their phenotypic antibiogram pattern).(TIF)Click here for additional data file.S1 File(DOCX)Click here for additional data file.S2 File(PDF)Click here for additional data file.


The overuse of antibiotics and the emergence of multiple-antibiotic-resistant pathogens are becoming a serious threat to health security and the economy. Reducing antimicrobial resistance requires replacing antibiotic consumption with more biocontrol strategies to improve the immunity of animals and humans. Probiotics and medicinal plants have been used as alternative treatments or preventative therapies for a variety of diseases caused by bacterial infections. Therefore, we reviewed some of the anti-virulence and bacterial toxin-inhibiting strategies that are currently being developed; this review covers strategies focused on quenching pathogen quorum sensing (QS) systems, the disruption of biofilm formation and bacterial toxin neutralization. It highlights the probable mechanism of action for probiotics and medicinal plants. Although further research is needed before a definitive statement can be made on the efficacy of any of these interventions, the current literature offers new hope and a new tool in the arsenal in the fight against bacterial virulence factors and bacterial toxins. Bacterial toxins are described as being products of microbial metabolism; they are one of the creative strategies that bacteria have established for survival. Generally, endotoxins and exotoxins should be differentiated. Bacterial endotoxins are the main component of the cell wall or outer membrane of Gram-negative bacteria. Endotoxins cannot be released when the bacteria are alive; when the cells die and dissolve or when the bacteria are destroyed artificially, they are released; thus, they are called endotoxins. The chemical composition of endotoxins includes phosphoric acid, polysaccharides and proteins. The main ingredient is lipopolysaccharide (LPS) , a heat-Staphylococcus aureus (S. aureus), Streptococcus and Bacillus tetanus [Pasteurella multocida and virulent Escherichia coli, can also produce exotoxins [Exotoxins are mainly soluble toxic proteins that are secreted by microbes into the surrounding media . Their t tetanus ,8,9. Somxotoxins ,11. Exotxotoxins . ExotoxiAlthough the exotoxins and endotoxins of bacteria are completely different in terms of structure and mechanism of action, they can cause varying degrees of serious damage to the body. In order to infect a host, bacteria must develop a mechanism to combat the host’s immune system so that they can survive the harsh environment in the host, so different virulence factors are expressed at different times during the infection. Virulence factors are microbial structures and biomolecules that enable pathogens to achieve colonization, invasion and persistence within susceptible hosts . Quorum-Drug-resistant infections have become an increasingly severe problem worldwide. Thus, alternative methods  are needed to address antimicrobial-resistant strains. This comprehensive review reveals the biocontrol approaches being employed to quench the pathogen QS systems, attenuate bacterial toxins and disrupt biofilm formation .Escherichia coli (STEC), inhibiting protein synthesis in cells with a nucleus and playing a role in the development of hemolytic uremic syndrome (HUS) and hemorrhagic colitis [stx2A expression was observed after a 2-h co-incubation of E. coli O157:H7 with Bacillus thermophilum, Lactobacillus rhamnosus, and Pediococcus pentosaceus. Their findings indicate that the downregulation of stx2A is possibly associated with the pH effect of organic acids from probiotic bacteria [E. coli strains can achieve the expression of Gb3 receptor mimics on their surfaces, which absorb and neutralize Stx2 in vitro. Piglets that receive oral administration of bacteria expressing Gb3 analogs are protected from lethal challenge by virulent STEC strains [Saccharomyces cerevisiae can be attributed to the elimination of secretory toxins, extracellular protease production, the stimulation of immunoglobulin A secretion, the hydrophobicity of cell surfaces, and autoaggregation ability [Saccharomyces boulardii (S. boulardii) in rat jejunum. They found a 120-kDa protein in a Saccharomyces boulardii-conditioned medium. This protein can inhibit cholera toxin-induced cAMP in intestinal cells. S. boulardii inhibited Cl− secretion in vitro via both Ca2+- and cAMP-mediated signaling pathways [S. boulardii CNCM I-745 was able to adhere cholera toxin to its cell wall, i.e., this was another mechanism to reduce the cholera toxin [Shiga toxins (Stx1 and Stx2) are the main factors responsible for the virulence of  colitis . Even thbacteria . Additio strains .  is a typical foodborne pathogen. This bacterium can generate over 16 toxins. These toxins can induce intestinal and histotoxic infections in animals and humans [B. coagulans can inhibit the expression and growth of the alpha toxin gene in C. perfringens type A and thus reduce apoptosis and cytotoxicity against HT-29 cells. Therefore, B. coagulans can be applied in the prevention and treatment of C. perfringens type A gut infections [It is well known that d humans . The prifections . As a fofections   production in lactobacilli and bifidobacteria could antagonize bacterial pathogens’ toxicity. The EPS produced by Bifidobacterium longum NB667 against SLO’s hemolytic activity were assessed using rabbit erythrocytes. EPS NB667 caused the greatest decrease in hemolysis in erythrocytes [Streptolysin O (SLO) is an exotoxin from hrocytes . Althoughrocytes   present a significant public health and safety risk due to their status as highly potent natural toxins. Both the BoNT serotype A complex and holo-toxin possess the capability to bind to and traverse intestinal epithelial cells, thus facilitating their systemic transportation through the bloodstream. According to Lam et al., probiotics such radation . Aggregatibacter actinomycetemcomitans (Aa) can induce inflammatory mediators in gingival epithelial cells. Then, Aa evades immune responses via internalization and the production of leukotoxin (LtxA) and cytolethal-distending toxin (CdtB) in non-phagocytic cells [Lactobacillus gasseri and Lactobacillus salivarius cell-free supernatants (CFSs), two genes were downregulated in a time-dependent manner, according to Nissen et al.’s analysis of the LtxA and CdtB levels [L. rhamnosus LR32 and Lactobacillus acidophilus LA5 CFS downregulated the transcription of CdtB and LtxA [ic cells ,33. WhenB levels . Anotherand LtxA .N-acyl-homoserine lactones (AHLs) are the primary signaling molecules serving as AIs in Gram-negative bacteria [Bacterial QS modulates gene expression based on bacterial population density via cell-to-cell communication . For Grabacteria ,38. AHLsbacteria . Microbibacteria , and thebacteria ,42. Disrbacteria . The degbacteria ,45. By ebacteria ,47. TherBiofilm formation is a crucial virulence factor. It is also a common phenomenon among microorganisms. Following biofilm formation, microbes can diffuse and colonize other environments . These bDirectly damaging targeted cells by producing antimicrobial metabolites and controlling adaptive mechanisms, such as biofilm formation, can induce antagonisms between bacteria and fungi or among bacteria. The QQ phenomenon allows for such an antagonistic approach. The co-development of secondary metabolites is vital in bacterial evolution. These secondary metabolites are capable of disrupting QS signaling molecules and attenuating other microorganisms’ virulence. The ability to disrupt QQ or QS signaling molecules may have evolved in QS bacteria in order to eliminate or repurpose their own QS signaling molecules or those of the microorganisms co-inhabiting competitive environments . MoleculLactic acid bacteria (LAB) have a significant effect on human life. In different countries, many types of LAB are utilized to produce traditional fermented food. Lactobacilli (types of LAB) are essential components of the normal intestinal microbiota of animals and humans. Primarily, with regard to the benefits of LAB, they can generate lactic acid and other substances . These substances can inhibit the growth of other bacteria ,61. LAB Lactobacillus plantarum ATCC 10241 has the ability to inhibit the production of AHLs, which are QS signaling molecules capable of inducing the virulence factors of Pseudomonas aeruginosa (P. aeruginosa) PA100 [P. aeruginosa PAO1 alone or in combination with zingerone [L. lactis, L. fermentum and L. rhamnosus) displayed a noteworthy decrease (p ≤ 0.05) in the auto-inducer AHL levels of P. aeruginosa PAO1 coupled with a decline in elastase activity [L. plantarum, and subsequently enclosed within alginate beads. L. plantarum strain SBR04MA produced the most significant outcomes. Within 9 h, all of the 6-HSLs were degraded by this strain [L. acidophilus 30CS cell extract was found to significantly inhibit the AI-2 activity of E. coli O157:H7, without effects on EHEC growth [L. plantarum Z057 against V. parahaemolyticus were found to be remarkable. It was observed that Z057-E could effectively impede EPS production as well as inhibit the QS signaling molecule AI-2 and extracellular protein secretions [Valdéz et al. have shown that a) PA100 . In a stingerone . In a stactivity   influenced the QS system in S. aureus by regulating the AI-2 activity [C. difficile were suppressed upon addition of the heat-treated supernatant L. fermentum Lim2. This was attributed to the suppression of agr genes [Many studies have been dedicated to interactions of lactobacilli with QS signals from Gram-positive bacteria besides than their ability to inhibit QS signaling in Gram-negative bacteria via interactions with various AHL types. The studies have focused on the inhibition of QS in t manner . In addiactivity   can induce serious infectious diseases. L. acidophilus inhibits lipase production and biofilm formation in S. aureus as reported by Sikorska et al. Their effects were mediated by the direct exclusion of cellular competition and by the production of bacteriocin-like inhibitors and short-chain fatty acids [L. fermentum TCUESC01, derived from cocoa seeds, effectively hindered biofilm formation of S. aureus. The mechanism of hindrance occurred via the suppression of two genes, icaA and icaR, which are known to significantly impact biofilm synthesis after the discharge of soluble molecules [L. johnsonii LJO02 (DSM 33828) and L. rhamnosus LR06 (DSM 21981) CFSs against MDR S. aureus (ATCC 43300) in two different media: an innovative animal derivative-free broth (TIL) and the conventional animal derivative-based MRS medium. All CFSs decreased the viability and metabolic activity of S. aureus. TIL was more effective in reducing S. aureus biofilm formation and stimulating LAB metabolism compared to MRS [L. sakei EIR/BG-1. These postbiotics could be employed as promising agents to prevent mastitis due to their antibacterial and antibiofilm activity against Streptococcus agalactiae ATCC 27956, Streptococcus dysgalactiae subsp. dysgalactiae ATCC 27957, and MRSA ATCC 43300 [An important human pathogen, methicillin-resistant ty acids . Melo etolecules . Squarzad to MRS . MastitiCC 43300   effect against S. mutans (ATCC 25175). Decreased cell viability and significant variations in biofilm formation were also proved using confocal laser scanning microscopy and scanning electron microscopy on glass coverslips and in vitro on human tooth surfaces. Furthermore, the expression of comA, comX, and gtfC was downregulated by sub-MIC values of the PMs without any significant inhibitory effects on growth [Streptococcus mutans growth, which is mainly due to peroxide production and organic acid generation; (3) the immunomodulatory effects resulting from the inhibition of IL-10 production and the induction of IFN-γ production; and (4) the downregulation of several Streptococcus mutans virulence genes, such as EPS-producing genes (gtf BCD and sacB), acid tolerance genes (atpD and aguD genes), and quorum-sensing genes (vicKR and comCD) [L. plantarum and L. acidophilus on multidrug-resistant E. coli from urine samples. In addition, Lb. plantarum and L. acidophilus strains reduced the ability of uropathogenic E. coli (UPEC) to develop biofilms by 39.63% and 56.3%, respectively. The variations in the gene expressions of csgA, crl, and csgD were significantly downregulated after separate treatment with L. plantarum and L. acidophilus suspensions [L. plantarum M.2 and L. curvatus B.67  on the formation of L. monocytogenes biofilm on various food contact surfaces. The higher sub-MICs  of both postbiotics considerably impeded biofilm formation at 30 °C for 24 h (p < 0.05) on silicone rubber, plastic, rubber gloves, and an MBEC™ biofilm device. However, the lower sub-MIC (1/16 MIC) did not have any significant inhibitory effects (p > 0.05). RT-qPCR results showed that the expression of several target genes associated with L. monocytogenes biofilm formation was downregulated by postbiotics. These target genes included those genes involved in QS (agrA), motility , and virulence , which significantly affect biofilm formation [As a chronic infectious disease with multiple factors, dental caries are initiated by bacterial biofilm formation, primarily by e agents . Emine en growth . It has d comCD) . Dawwam pensions . Hossainormation . The lacOver the past decade, an incredible amount of research has been conducted to identify plants with anti-QS properties and biofilm inhibition activity. Phytochemicals can regulate bacterial AHL synthesis and in turn suppress QS. Various natural extracts are thought to inhibit QS by disrupting AHL activity via competition with AHLs due to their structural similarity and/or by accelerating the degradation of the LuxR/LasR receptors of AHL molecules .The chemical classes from the investigated anti-virulence medicinal plants mainly include phenolic derivatives, terpenoids, and alkaloids.Agrobacterium tumefaciens QS signaling receptor TraR in Escherichia coli cells when present at millimolar concentrations. Additionally, this particular flavone demonstrates inhibitory effects on biofilm formation dependent on QS in P. aeruginosa PAO1 at micromolar concentrations [V. harveyi and E. coli [Phenolic compounds belong to a large group of secondary plant metabolites. Phenol (the simplest phenolic compound) is an aromatic ring with a single hydroxyl group. Polyphenols are composed of two or more such phenolic units and exhibit a broad structural diversity. Flavonoids belong to the polyphenol family and are water-soluble polyphenolic molecules with 15 carbon atoms. It has been reported that baicalein, a type of flavone, exhibits the potential to enhance proteolysis of the trations ,83. It htrations . This is E. coli . Salmonella typhimurium via the modulation of luxS and diA gene expression levels [Listeria monocytogenes [Burkholderia cepacia [Eikenella corrodens [As an essential constituent of rich tea, epigallocatechin exhibits antibiofilm properties against n levels . Epigallytogenes , Burkhol cepacia , and Eikorrodens .Escherichia coli O157:H7 [P. aeruginosa PAO1, P. aeruginosa PAF79, and Aeromonas hydrophila WAF38 [P. aeruginosa strain can be effectively inhibited by quercetin, thus demonstrating antibiofilm properties. Additionally, quercetin at a concentration of 16 μg/mL has the ability to decrease the expression levels of rhll, rhlR, lasl, and lasR [LasR in P. aeruginosa to inhibit QS circuitry [Quercetin, which is widely distributed in fruits and vegetables, has diverse and multiple effects against QS depending on the tested bacterial strains. Quercetin also shows antagonistic effects on bacterial signaling. In addition, the suppression of biofilm formation by quercetin was found in  O157:H7 . Quercetla WAF38 . Accordiand lasR . Quercetircuitry .S. aureus by reducing its motility, spreading ability, and production of EPS [S. aureus sortaseA (SrtA) can be inhibited by kaempferol at a concentration of 64 μg/mL [P. aeruginosa while not affecting bacterial growth. Moreover, this substance can lower the expression of specific QS-controlled genes  in P. aeruginosa PAO1 [Aeromonas. Dong et al. found that genistein can inhibit biofilm formation and aerolysin generation, which is dose-dependent. When co-cultured with genistein, the transcription of QS-related genes (ahyI and ahyR) and the aerolysin encoding gene (aerA) were drastically downregulated [Aeromonas hydrophila [Other flavonoids have also been found to exhibit significant anti-QS properties. For instance, a study conducted by Chemmugil et al. revealed that morin effectively impeded the formation of biofilms in n of EPS . Up to 864 μg/mL . Conversosa PAO1 . As an iosa PAO1 . Aerolysegulated . In addidrophila ,99.P. carotovorum ssp. brasiliense and P. aroidearum and thus changed the expression of their bacterial virulence factors. Although the expression of QS-related genes increases over time in control treatments, the exposure of bacteria to non-lethal concentrations of cinnamic acid or SA can suppress the expression of QS genes  and luxS (a component of the AI-2 system)), expI, and exp R [P. aeruginosa P1 to encourage the outer membrane’s detachment. The expression of the major genes (LPxB and LPxC) in LPS biosynthesis can also be downregulated by CA [As natural antioxidants in plants, phenolic acids (PAs) have an aromatic ring with methoxy or OH groups. The number of methoxy or OH groups determine their diversity. Their antibacterial activities are associated with their chemical structures ,101. Josnd exp R . Chloroged by CA . Vibrio alginolyticus, a pathogen that affects fish. VA was found to significantly reduce the ability of V. alginolyticus to form biofilms, its mobility, and its production of exotoxins such as protease and exopolysaccharide. In addition, VA downregulated the expression of genes  associated with biofilm formation and virulence in V. alginolyticus when used at sub-inhibitory concentrations [A. hydrophila strains. Additionally, RA has the ability to decrease the expression of virulence genes such as lip, ahyB, ahh1, and aerA [Liu and colleagues conducted a study on the effects of vanillic acid (VA) against trations . Kannan and aerA .A. tumefaciens on abiotic (polystyrene and nitrocellulose) and plant root surfaces and can inhibit the virulence traits . One mechanism for antibacterial and subsequent biofilm inhibition may be attributed to iron chelation via tannic acid because of galloyl groups. The reduced expressions of chvE and dnaK may be the other mechanism [E. coli [S. aureus [S. aureus are mainly related to the production of protein immunodominant staphylococcal antigen A (IsaA). IsaA is a putative lytic transglycosylase that is capable of cleaving the β-1,4-glycosidic bonds between N-acetylglucosamine (GlcNAc) and N-acetylmuramic acid (MurNAc) in the peptidoglycan layer. Cleaving the peptidoglycan layer can reduce the biofilm thickness of S. aureus [sdiA and srgE) in Salmonella and the production of violacein by Chromobacterium violaceum [Salmonella is not capable of synthesizing AHLs on its own, it can perceive AHL signals from other microorganisms via SdiA (a LuxR homolog) in Salmonella. Salmonella can also control a broad range of traits [Salmonella Paratyphi A and Salmonella Typhi regulated via QS at a concentration of 400 μg/mL. Tannic acid demonstrates the ability to inhibit QS and associated virulence factors without causing harm or exerting continuous pressure on the viability of the pathogen [Salmonella sp. has the SdiA and luxS/AI-2 QS systems [Tannins can affect toxin production and are significant for bacterial pathogenicity and virulence. Jailani et al. reported that polyphenol tannic acid can efficiently inhibit the biofilm formation and planktonic growth of echanism . Tannic [E. coli  and S. a. aureus . Its ant. aureus  , Cannabis sativa L. (Cannabaceae), and Cinnamomum camphora (L.) [Acinetobacter baumannii (A. baumannii). Its mechanism of action against A. baumannii involves disrupting quorum sensing, impeding biofilm development, and inhibiting bacterial adhesion. Linalool effectively hinders the formation of biofilms by dispersing them and preventing their growth [S. aureus treated with geraniol decreases at 1 mg/mL, and its biofilm formation shows a decrease at concentrations between 0.5 and 4 mg/mL [MRSA via inhibitory effects on biofilms and anti-virulence activity against the main virulence factors  [Streptococcus mutans and reduces the expression of gtf genes [Terpenoids are derived from the condensation of isopentane or isoprene units and mainly include sesquiterpenoids, monoterpenoids, and diterpenoids. Linalool is a monoterpenoid with a tertiary alcohol group, which is derived from many plants, including J. Presl (Lauraceae), ora (L.) ,115,116.r growth . Some mo 4 mg/mL . The ant lipase) . With retf genes . S. pyogenes and S. mutans, respectively. This monoterpenoid exhibits a dose-dependent response, inhibiting the adhesion properties of S. pyogenes biofilms while also downregulating vicR gene expression in S. mutans, ultimately leading to reduced acid production [E. radiate and E. globules also show anti-QS properties. 1,8-cineole is the major component of E. globulus oil and is known as eucalyptol (63.81%). 1,8-cineole in E. globulus oil and limonene (68.51%) in E. radiata oil suppress the QS phenomenon by inhibiting QS-regulated violacein pigment production in bacteria without any effects on their growth [Dorema aucheri Bioss. and Ferula asafoetida L. EOs at 25 μg/mL exhibit anti-QS activity against P. aeruginosa. Ferula completely inhibited the violacein production of C. violaceum, while Dorema only reduced the violacein production of C. violaceum to some extent at the same concentration. Pyoverdine, elastase, pyocyanin, and biofilm production were reduced under treatments of Ferula oil. Dorema oil diminished elastase and pyoverdine production while not affecting biofilm and pyocyanin production [Monoterpenes and sesquiterpenes, as well as their oxygen derivatives, are the most conventional types of citrus essential oils (EOs) . The widoduction . The EOsoduction . EOs fror growth . Dorema oduction .S. epidermidis. It was found that berberine has bacteriostatic effects on S. epidermidis. Berberine can inhibit the formation of S. epidermidis biofilms at sub-minimal inhibitory concentrations. Berberine suppresses bacterial metabolism at concentrations of 15–30 μg/mL and can exhibit antibacterial effects and significantly inhibit the biofilm formation of S. epidermidis ATCC 35984 and its clinical isolate strain SE243S at moderate concentrations of 30–45 μg/mL [Alkaloid berberine is another compound exhibiting antibiofilm activity. It binds to EPS-associated amyloid proteins to affect the biofilm formation of 45 μg/mL . The medThe threat of antimicrobial resistance has motivated the scientific community to find effective solutions. Therefore, the biocontrol of pathogen virulence has been seen as a promising alternative approach which aims to inhibit the production or activity of vir-ulence factors and not affect general bacterial growth. The current studies of probiotics or natural plants are mainly aimed at the prevention of bacterial adhesion or biofilm formation, the interruption or inhibition of the work of bacterial secretion systems, and the downregulation of virulence. Other strategies include reducing or blocking quorum sensing, and via global or specific regulators, gene expression regulation was performed. Here, we review the possible mechanisms and research progress of probiotics  in neutralizing bacterial toxins such as Shiga toxin, cholera toxin, alpha toxin, streptolysin O, and botulinum neurotoxin. These probiotics exhibit potential antibacterial activity while also disrupting the expression of genes associated with the quorum sensing system of pathogenic bacteria and influencing biofilm formation. The chemical classes from the investigated anti-virulence medicinal plants mainly included phenolic derivatives, terpenoids, and alkaloids. Relevant secondary metabolites from medicinal plants have biofilm-inhibitory activity and anti-quorum sensing (QS) characteristics. As an emerging therapeutic strategy, the development of biocontrol therapy still faces several challenges. The main limitations of studies on biocontrol of bacterial toxins and virulence include the paucity of human or animal studies; their ideal dose; the duration of supplementation; and the durability of their beneficial effects as well as their safety profile in the treatment of pathogenic bacteria.It is clear that medicinal plant resources and probiotics are abundant. However, many aspects have not been studied. It should also be recognized that the working modes of many effective inhibitors remain unclear. Therefore, the study of their bioactive mechanisms can be taken as a future research direction.The determination of whether probiotics and medicinal plants can concurrently re-duce both pathogens and toxins produced by the pathogens.The development of probiotic-based oral vaccines to protect animals against contamination by toxins.The determination of whether the anti-toxin effects of probiotics and medicinal plants and extracts in vitro can be duplicated in vivo, especially in humans.To promote further progress, future studies need to solve one or more of the following aspects of the biocontrol of bacterial toxins:


Plasmodium species, is a promising malaria vaccine candidate. CelTOS is essential for parasite invasion into mosquito midgut and human hepatocytes, thereby contributing to malaria transmission and disease pathogenesis. This study explores the genetic diversity, polymorphisms, haplotypes, natural selection, phylogenetic analysis, and epitope prediction in the full-length Plasmodium knowlesi CelTOS gene in clinical samples from Sarawak, Malaysian Borneo, and long-term laboratory strains from Peninsular Malaysia and the Philippines. Our analysis revealed a high level of genetic variation in the PkCelTOS gene, with a nucleotide diversity of π ~ 0.021, which was skewed towards the 3’ end of the gene. This level of diversity is double that observed in PfCelTOS and 20 times that observed in PvCelTOS from worldwide clinical samples. Tests of natural selection revealed evidence for positive selection within clinical samples. Phylogenetic analysis of the amino acid sequence of PkCelTOS revealed the presence of two distinct groups, although no geographical clustering was observed. Epitope prediction analysis identified two potential epitopes (96AQLKATA102 and 124TIKPPRIKED133) using the IEDB server and one epitope (125IKPPRIKED133) by Bcepred server on the C’ terminal region of PkCelTOS protein. Both the servers predicted a common epitope region of nine amino acid length (IKPPRIKED) peptide, which can be studied in the future as a potential candidate for vaccine development. These findings shed light on the genetic diversity, polymorphism, haplotypes, and natural selection within PkCelTOS in clinical samples and provide insights about its future prospects as a potential candidate for P. knowlesi malaria vaccine development.The cell-traversal protein for ookinetes and sporozoites (CelTOS), expressed on the surface of ookinetes and sporozoitesin  Anopheles mosquito is infected with Plasmodium spp. and injects the parasite into the human bloodstream [Plasmodium species, nine have been identified to cause infection in humans. Plasmodium vivax, P. falciparum, P. ovale curtisi, P. ovale wallikeri, and P. malariae are human malaria parasites well-known to cause malaria in humans [Plasmodium parasites of simian origin, e.g., P. knowlesi, P. simium, P. brasilianum, and P. cynomolgi, have also been reported to infect humans [P. falciparum and P. vivax. P. falciparum exhibits the highest virulence and is responsible for the highest mortality, especially in the African continent [P. vivax is the most prevalent species [P. knowlesi has been discovered to cause human infection, as reported from several Southeast Asian countries, particularly in Malaysian Borneo [P. knowlesi malaria has become a significant public health issue in the region [Anopheles hackeri, was first identified as the vector carrying P. knowlesi in Malaysia [Anopheles were reported to transmit the parasite from macaques to humans [Macaca fascicularis) and pig-tailed macaques (Macaca nemestrina) [Presbytis melalophos, or banded leaf monkeys, are also reported to harbor P. knowlesi naturally [P. knowlesi malaria can cause severe and complicated malaria cases in humans, with symptoms similar to other malaria, including fever, headache, vomiting and diarrhea, acute respiratory distress syndrome, and multi-organ failure like kidney failure [P. knowlesi can lead to a swift rise in parasitemia, potentially resulting in death [P. knowlesi cannot be accurately identified through microscopy because its early trophozoites have the same features as P. falciparum, and its later stages resemble the band-form of trophozoites of P. malariae [P. knowlesi has been witnessed in humans over the last decade, which is presumably due to the changes in the environment like large-scale deforestation and land exploration. This might have played a key role in increasing human exposure to anopheline vectors [Malaria is a life-threatening disease predominant in most tropical and sub-tropical countries. The disease is transmitted when a female odstream . The illodstream . Out of n humans ,3,4,5. It humans ,7. Amongontinent . In many species . The simn Borneo ,12,13,14e region ,13,15. TMalaysia . Later oo humans ,13,15. Po humans ,11,17. Aestrina) . Presbytaturally . P. know failure ,20. The in death . P. knowmalariae ,22. PCR malariae . In Mala vectors . Thus, tP. vivax have revealed that the sporozoite-expressed genes exhibit a higher degree of conservation than the merozoite genes [Mosquirix targeting P. falciparum circumsporozoite protein (CSP), is the world’s first malaria vaccine to receive approval from WHO and is in current use [P. falciparum and P. vivax in P. knowlesi to develop an efficacious vaccine candidate for knowlesi malaria. Various studies have characterized merozoite invasion genes in P. knowlesi, analyzing their genetic diversity and population structure [P. knowlesi.The prime challenge of developing a vaccine can be attributed to the genetic diversity found in the field samples within the potential vaccine candidates leading the vaccine to be non-efficacious ,25. Studte genes . The RTSrent use ,28; howetructure ,17,25,29tructure  have notPlasmodium’s life cycle. It is an essential micronemal protein that enables the parasite to cross host cell barriers [P. berghei CelTOS gene can significantly reduce the parasite’s ability to infect both mosquito host and human liver, almost eliminating the ability of the parasite to pass through host cells successfully [P. berghei, establishing its potential as a vaccine candidate [P. berghei and P. falciparum parasites that expressed PfCelTOS in Anopheles gambiae mosquitoes [E. coli-expressed PfCelTOS-based vaccine is already in the Phase 1 trial [Cell-traversal protein for ookinetes and sporozoites, briefly known as CelTOS, is a surface protein expressed in sporozoites and ookinetes in the sporogonic stages of barriers ,32,33,34barriers . A studyessfully . Anotheressfully . Antibodessfully , and immandidate . Ex vivoandidate . In anotandidate . Furtherandidate . This efsquitoes , suggestsquitoes . A recom 1 trial .CelTOS genetic diversity in P. vivax and P. falciparum, while none have investigated P. knowlesiCelTOS. Thus, in the present study, we aimed to find the genetic diversity, polymorphisms, and natural selection operating on full-length PkCelTOS gene sequences from 34 samples . We also conducted a phylogenetic analysis to find the relationship between PkCelTOS amino acid sequences and their orthologs in other species. We predicted potential epitopes of PkCelTOS, which might induce an immune response during the sporozoite and ookinete stages of the parasite. Since our study is the first to examine CelTOS sequences from clinical samples of this zoonotic parasite, the study findings will contribute to the rational design and formulation of ookinete and sporozoite stage vaccines that are effective against P. knowlesi.Prior studies have explored PkCelTOS full-length gene (from 1 to 558 nt) sequences  were obtained from the public database European Bioinformatics Institute  [PkCelTOS gene sequences were included for analysis. The sequences were aligned using the CLUSTAL-W program within the MegAlignLasergene v 7.0 software . SignalP-5.0 prediction software  was used to predict the signal peptide for the full-length PkCelTOS protein [A total of 34 ch 2023) . These sch 2023) . Only hi protein .PkCelTOS derived from clinical samples in Malaysian Borneo and long-term lab strains from Peninsular Malaysia and Philippines . Phylogenetic analysis was conducted in MEGA 5.0 software utilizing Maximum Likelihood (ML) method on the basis of the Poisson correction model with 1000 bootstrap replicates [P. coatneyi (PCOAH_00055260), P. cynomolgi (PCYB_144300), P. vivax (PVX_123510), and P. falciparum (PF3D7_1216600) were also added to investigate the evolutionary relationship between the CelTOS protein of P. konwlesi along with its closest members in the genus Plasmodium.Phylogenetic analysis was carried out to establish the intra-species relationship between amino acid sequences of plicates . OrtholoPkCelTOS full-length gene was determined using DnaSP v6.12.03 software [Nucleotide diversity (π) in the software . Other pPkCelTOS at the intra-species level, the neutrality tests of Tajima’s D, Fu, and Li’s F*and D* were computed using DnaSP software. Tajima’s D statistic yields a value of zero under conditions of neutrality. A negative Tajima’s D result determines population expansion, while a positive and significant Tajima’s D result signifies positive selection or balancing selection. Using the DnaSp software, the results of Tajima’s D were also shown graphically (with a window length of 50 and step size of 10 bp). Positive and significant values of Fu and Li’s F* and D* suggest that the population may have undergone a contraction. Conversely, an excess of singletons or negative values can indicate population expansion. In addition, the method of Nei and Gojobori was utilized for calculating the rate of non-synonymous substitution per non-synonymous site (dN) and the rate of synonymous substitution per synonymous site (dS) [To determine the natural selection within ite (dS) .PkCelTOS gene sequences with their closely related ortholog sequences in P. coatneyi (PCOAH_00055260), P. cynomolgi (PCYB_144300) and P. vivax (PVX_123510).The robust McDonald and Kreitman (MK) test was carried out to examine natural selection at the inter-species level, utilizing DnaSP v6.12.03 software using 34 PkCelTOS gene using the Datamonkey Web Server (https://www.datamonkey.org/ accessed on 1 July 2023) [Codon-based analysis of selection was performed on the ly 2023) . The analy 2023) .PkCelTOS sequence of P. knowlesi. The epitope prediction was conducted using two servers; the IEDB Analysis resource (http://tools.immuneepitope.org/bcell), with the Emini Surface Accessibility Prediction model to identify B-cell epitopes on the surface [https://webs.iiitd.edu.in/raghava/bcepred/index.html) [P. knowlesi and P. vivax are phylogenetically close and several studies have shown the importance of conserved cross-species vaccine candidates [P. knowlesi, this study examined the likelihood of epitope conservation with P. vivax.B-cell epitopes are specific sites on a protein that can be recognized by the antigen-binding sites present on the immunoglobulin molecules. These epitopes play a crucial role in the formulation of peptide-based vaccines, disease detection, and allergy studies . In this surface , and theex.html) , using tex.html) . The metex.html) . Since Pndidates ,49,50. TPkCelTOS protein  of 0.954 ± 0.016. P. knowlesi reference H-strain. The nucleotide and amino acid polymorphisms of the clinical strains with respect to the reference H-strain, along with nucleotide positions, were illustrated in Analysis of 34  0.00105 , which iPkCelTOS gene sequences obtained from 34 sequences showed overall Tajima’s D value to be D = 1.52705, p > 0.10, which was not significant  but not significant.Analysis of the natural selection of nificant . Tajima’ificance . In addiPkCelTOS gene may be undergone positive selection when tested with CelTOS orthologs in P. vivax, P. cynomolgi and P. coatneyi , while MEME detected one (171) site undergoing positive selection. Analysis conducted using the SLAC method 1 (111) site detected to be under positive selection. Remarkably, both FUBAR and MEME concurred on amino acid position 171, undergoing positive selection. Likewise, both FUBAR and SLAC concurred on a common site 111 to be under positive selection. In contrast, FUBAR identified four negatively selected sites , while SLAC identified two sites (73 and 87) undergoing strong negative selection. Notably, both methods agreed on amino acid positions 73 and 87 experiencing negative selection. A plot depicting positions for positive and negative selection of the PkCelTOS protein sequence generated using SLAC is shown in The departure of neutrality of the P. knowlesi CelTOS identified two surface-exposed epitopes (96AQLKATA102 and 124TIKPPRIKED133) towards the C’ terminal region of the protein , and hepatocytes [P. vivax and P. falciparum, but it has not yet been explored in Plasmodium knowlesi. The present study is a pioneer in investigating the genetic diversity and polymorphisms within the PkCelTOS gene in clinical samples from Malaysia, as well as predicting the B-cell epitopes that can be further studied to develop sporozoite vaccines.The life-cycle of atocytes ,31,51,52atocytes ,31,51,52atocytes . Thus, aPkCelTOS, it was observed that it exhibits the highest sequence identity with PvCelTOS, which may indicate the ongoing adaptation for invading the physical barriers of the human host’s immune system. The signal peptide region was conserved among all clinical samples of P.knowlesi, indicating that it may play a critical role in directing the newly synthesized protein to its intended destination, i.e., sporozoites and ookinetes. The results of a phylogenetic analysis by ML method did not show any geographical clustering as both the clusters had laboratory lines originating from Peninsular Malaysia and the Philippines, together with clinical samples from Sarawak, Malaysian Borneo. Similar results were also found by Assefa et al., 2015 using the same set of sequences [P. knowlesi genome sequences from clinical samples [PkCelTOS is expressed in sporozoite and ookinete surfaces and may not be exposed to human immune pressure. PkCelTOS was found to be phylogenetically most related to PvCelTOS, its ortholog infecting humans. Among the simian malaria parasites, P. coatneyi CelTOS is closer to PkCelTOS. The overall nucleotide diversity of PkCelTOS (π = 0.02111 ± 0.00105) was higher than the nucleotide diversity of PfCelTOS (π = 0.01001 ± 0.00036) worldwide samples [PvCelTOS (π = 0.00141 ± 0.00014) [PkCelTOS was also found to be higher than other sporozoite stage vaccine candidates, e.g., PkTRAP (π = 0.00908 ± 0.0006) [By aligning 34 full-length amino acid sequences of equences , except equences , PkRhopHequences , MSP4 [2equences , MSP1P [equences , nbpxa [equences  along wi samples  has show samples  and 20 t0.00014) . The nuc 0.0006)  and mero 0.0006)  but lowe 0.0006) . The lowPkCelTOS gene may be under positive selection; however, the results were statistically not significant, which may be due to a smaller sample size. However, dN-dS results are found to be negative but not significant. A higher number of non-synonymous mutations than synonymous mutations was found in the McDonald and Kreitman (MK) neutrality test between PkCelTOS orthologs. Codon-based site-by-site selection analyses in Datamonkey identified five potential sites which could be under positive natural selection and four sites under purifying selection. This is probably due to protein folding, and the sites under positive selection might be exposed to host immune pressure, while the negatively selected sites are under functional constraints and not exposed to the host. These findings may indicate that the gene in question is undergoing long-term differential selective pressure from the human immune system, similar to the conclusions of PfCelTOS [P. knowlesi and P. vivax. Nevertheless, further sequence analysis involving a more extensive sample size would be necessary to substantiate the presence of the epitope.Natural selection tests  yielded positive results, implying that the PfCelTOS . Both IECelTOS gene in Plasmodium knowlesi, a malaria parasite prevalent in Southeast Asia. The high degree of nucleotide diversity observed in the PkCelTOS gene suggests this protein may be under selective host immune pressure. However, the identification of two potential epitopes on the PkCelTOS protein provides a promising avenue for the development of a vaccine that can stimulate an effective immune response. The phylogenetic analysis of PkCelTOS revealed two distinct groups, indicating that there may be genetic differences between Plasmodium populations in different regions of Malaysia. The absence of geographic clustering suggests that the genetic variation observed in the PkCelTOS gene is likely the result of selective local pressures rather than geographic isolation. In conclusion, this study highlights the need for continued surveillance of PkCelTOS genetic diversity in different regions. The findings of this study offer valuable knowledge for the rational design of peptide-based vaccines against P. knowlesi malaria. However, further functional studies, as well as genetic studies with a higher number of samples, are needed to validate the effectiveness of a PkCelTOS-based vaccine.To conclude, this present study provides a valuable understanding of the genetic diversity and polymorphism across the full-length gene of


Enterococcus hirae is a rare pathogen in human infections, although its incidence may be underestimated due to its difficult isolation. We describe the first known case of E. hirae infective endocarditis (IE), which involves the mitral valve alone, and the seventh E. hirae IE worldwide. Case presentation: a 62-year-old male was admitted to our department with a five-month history of intermittent fever without responding to antibiotic treatment. His medical history included mitral valve prolapse, recent pleurisy, and lumbar epidural steroid injections due to lumbar degenerative disc disease. Pre-admission transesophageal echocardiography (TEE) showed mitral valve vegetation, and Enterococcus faecium was isolated on blood cultures by MALDI-TOF VITEK MS. During hospitalization, intravenous (IV) therapy with ampicillin and ceftriaxone was initiated, and E. hirae was identified by MALDI-TOF Bruker Biotyper on three blood culture sets. A second TEE revealed mitral valve regurgitation, which worsened due to infection progression. The patient underwent mitral valve replacement with a bioprosthetic valve and had an uncomplicated postoperative course; he was discharged after six weeks of IV ampicillin and ceftriaxone treatment. Enterococcus hirae is a known pathogen of some animal species, causing endocarditis in chickens, diarrhea in rats, mastitis in cattle, intra-abdominal infection in cats, and sepsis in birds [E. hirae septicemia [in birds . The firpticemia . Since tpticemia .Enterococci are a frequent cause of infection in humans, only 0.4% to 3.03% are reported to be from E. hirae species; nevertheless, these data may be underestimated due to its difficult isolation, and its identification could increase using MALDI-TOF: a diagnostic system with a higher sensitivity level compared to previous ones [Although ous ones ,3.E. hirae IE.We describe a case of mitral valve infective endocarditis (IE) in a 62-year-old male reviewing the previously reported cases of A 62-year-old male was admitted to our department for a five-month history of intermittent fever without responding to antibiotic treatment alongside weight loss, sweating, coughing, and fatigue.His medical history included mitral valve prolapse causing severe regurgitation, recent pleurisy, and lumbar epidural steroid injections due to lumbar degenerative disc disease.Enterococcus faecium was identified by MALDI-TOF VITEK MS on blood cultures. According to the 2023 Duke-International Society for Cardiovascular Infectious Diseases IE Criteria, with two major clinical criteria and two minor clinical criteria, the patient was diagnosed with infective endocarditis and was, therefore, admitted to our Department [Twelve days before admission, a TEE performed for persistent fever showed one vegetation on the mitral valve posterior leaflet measuring 4 × 6 mm, and At admission, the patient was conscious with a body temperature of 38.5 °C, a pulse rate of 112/min, a respiratory rate of 20/min, oxygen saturation of 98% on room air, and blood pressure of 119/76 mm Hg. Laboratory investigation revealed a white cell count of 4900/microliters, a C-reactive protein of 37.8 mg/L, and procalcitonin of 0.173 μg/L.On day 1, blood cultures were taken, and intravenous therapy with ampicillin 2 g q4h and ceftriaxone 2 g q12h was initiated, according to antibiotics sensitivity. A good clinical response was obtained, and the temperature was resolved after 24 h.Enterococcus hirae (using MALDI-TOF Bruker Biotyper) with an antibiotic sensitivity profile  the same as the one obtained on E. faecium identified before admission  and renal fascia thickening. The colonoscopy showed no pathologic findings.On day 17, a TEE was repeated and showed a dilated left ventricle and mitral posterior leaflet prolapse, determining severe regurgitation. Moreover, two vegetations attached to the posterior mitral leaflet of 5 × 6 mm and 6 × 7 mm, respectively, were identified. Therefore, the patient underwent mitral valve replacement with a bioprosthetic valve after four weeks of antibiotic treatment. Mitral valve leaflets cultures and blood cultures taken after surgery were negative.The patient had an uncomplicated postoperative course and was discharged after a six-week IV ampicillin and ceftriaxone course. The timeline of clinical events is depicted in Enterococcus hirae without limits. Furthermore, all references listed were hand-searched for other relevant articles.Online research without language restrictions was conducted using PubMed and SCOPUS. This research was performed by combining the terms endocarditis and The PubMed research identified 19 publications; their meticulous analysis led to 15 eligible articles, and no other articles (not present in PubMed) were found through the SCOPUS research. Overall, six articles describing the history of six patients, published between 2002 and 2020, were further evaluated.Data regarding clinical characteristics, therapy, and the outcomes of the 6 patients were retrieved alongside that of our present case, which is analytically shown in Enterococci are the third cause of infective endocarditis, after staphylococci and streptococci [E. hirae is a member of the Enterococcus genus and is a well-documented cause of infection in animals  [Infective endocarditis (IE) is a life-threatening disease associated with high mortality and severe complications. ptococci . E. hirae birds) . Otherwie birds) .E. hirae is an important cause of endocarditis in broilers [E. hirae IE in humans documented worldwide. Moreover, among the six previously described cases, five involved the aortic valve only, and one involving both the aortic and mitral valve. This is the first documented case of E. hirae IE involving only the mitral valve.Although broilers , our casIn the present case, the patient was affected by mitral valve prolapse with severe valve regurgitation. Predisposing risk factors for cardiac pathology were also present in five out of six previous patients, such as a history of coronary artery disease with percutaneous coronary intervention, aortic valve replacement with a bioprosthetic valve, cardiac arrhythmia with surgical ablation and patent foramen ovale, severe aortic regurgitation, severe aortic stenosis and the bicuspid aortic valve ,6,7,8,9.E. hirae is a common pathogen in animals, it is not documented as a clear E. hirae exposure in the previously reported cases nor in our one before symptom onset [Although om onset ,7,8,9,10E. hirae IE had a history of gastrointestinal disease ; theref adenoma . In the E. faecium, whereas E. hirae was correctly identified to the species level through the three blood cultures drawn on day 1 of hospitalization in our department. In both cases, species identification was performed by mass spectrometry analysis MALDI-TOF MS .The first blood culture taken before admission to our department identified E. hirae were also encountered in the first two documented cases of E. hirae IE, in which a rapid ID 32 Strep system misidentified the genus as E. durans and E. gallinarum. In both cases, E. hirae was subsequently and correctly identified using sodAint gene sequencing [Lactococcus garvieae, initially identified as E. hirae IE, was described as well [Difficulties in the identification of quencing ,6. A cas as well .E. hirae IE cases, species identification was performed by MALDI-TOF MS in cases four, five, and six, while it was unknown in the third.In the other known Enterococci with a high level of species-level discriminatory power [E. hirae incidence may have been the consequence of the limited power of other diagnostic systems; indeed, the identification of E. hirae increased after the development of MALDI-TOF MS [E. faecium.MALDI-TOF MS proved to be an accurate system for the identification of both usual and unusual ry power . The lowI-TOF MS . Both thI-TOF MS , as corrE. hirae IE, five of which needed valve replacement surgery. The first two patients relapsed after treatment with ampicillin or amoxicillin and gentamicin, followed by rifampicin for six weeks. In the first case, vancomycin and gentamicin were administered for six weeks after relapse; in the second, the initial therapy was successfully repeated [All seven patients survived repeated ,6. The trepeated . Ceftriarepeated ,9. The srepeated . In our The European Society of Cardiology guidelines recommend a prolonged administration (up to 6 weeks) of synergistic bactericidal combinations of two cell wall inhibitors (ampicillin or penicillin and ceftriaxone for penicillin-susceptible strains) or one β-lactams with aminoglycosides for the management of enterococcal IE .E. hirae to antimicrobial agents suggests that this species is more similar to E. faecalis than E. faecium [E. faecalis IE [E. hirae IE, reducing the risk of renal failure, which is typically associated with gentamicin administration [The susceptibility of  faecium . Treatmecalis IE ,18. TherE. hirae is a potential pathogen in severe human infections such as endocarditis; its incidence is expected to increase thanks to MALDI-TOF MS, an accurate system with a higher species-level discriminatory power than previous ones. Treatment with ampicillin and ceftriaxone for enterococcal IE is associated with a good prognosis and a low risk of side effects and could be a good therapeutic choice in E. hirae IE.In summary,


Klebsiella pneumoniae (K. pneumoniae) has been commonly associated with human nosocomial infections and has recently gained special attention as a clinically important pathogen in companion animals   has emerged as a new multidrug-resistant K. pneumonia clone worldwide in both humans and animals   –4. In re animals . There a animals , Germany animals  and 2018 animals . K. pneufections . In Sout's study . Recent K. pneumonia enteritis in a kennel of Bordeaux mastiffs, resulting in septicemia and death, has been reported in a previous study  and has been used in epidemiological investigations . By compK. pneumoniae isolates were isolated from the blood (KP-B) and fecal (KP-F) samples, and the urine samples were negative for bacterial culture. The isolates were identified using MALDI-TOF, and antibiotic susceptibility of the isolates was determined by the Kirby-Bauer Test disc-diffusion method as recommended by the Clinical and Laboratory Standards Institute for a consensus interpretive criterion  for evaluation of a 1-month history of diarrhea, fever, lethargy, and anorexia in September 2022. Blood, urine, and fecal samples collected from the dog were submitted to NosVet Laboratory  to isolate the causative agent and antibiotic resistance test. Two riterion .For WGS, genomic DNA was extracted from pure cultures of the KP-B and KP-F using the MagNA Pure 96 DNA and Viral NA Small Volume Kit on the MagNA Pure 96 instrument  according to the manufacturer's instructions. Sample DNA concentrations were determined using a Qubit BR dsDNA assay kit , and DNA (0.2 ng/μl)were used for the library preparation using the Illumina Nextera XT DNA Library Prep Kit  as previously described . The libhttps://sourceforge.net/projects/bbmap) (Q > 20 and minimum length >50), and trimmed reads were de novo assembled using the SPAdes 3.15.5 (https://www.geneious.com/). The assembled contigs with coverage of <5 × and size below 300 bases were removed. To confirm species identification, the 16S rRNA regions in the assembled contigs of the isolates were predicted by barrnap , and the nearest-neighbor species with >99% identity were first searched using the BLASTn on the NCBI database (https://blast.ncbi.nlm.nih.gov/Blast.cgi) with the default parameters for each 16S rRNA sequence. MLST 2.0 (Multi-Locus sequence typing) was used to determine the sequence type of the isolates. The presence of acquired antimicrobial resistance genes and chromosomal mutations in the gyrA, gyrB, parC, and parE genes were determined using ResFinder 4.1 (https://cge.food.dtu.dk/services/ResFinder-4.1/) with settings of a threshold of 90%, and a minimum length of 60% with the assembled contigs.The raw reads were adapter-trimmed for known Illumina adapters and quality-trimmed with Bbduk (s 3.15.5  with itsK. pneumoniae ST 307 from cats and dogs (n = 37) were downloaded from BV-BRC (https://www.bv-brc.org/) and BIGSdb-Pasteur (https://bigsdb.pasteur.fr/). WGS of two K. pneumoniae ST 307 isolates (KP 44 and KP 45) from dogs referred to the Veterinary Medical Teaching Hospital of Konkuk University were also included for the subsequent WGS analysis. Sample information of the genomes is listed in K. pneumoniae ST 307 isolates including our isolates were compared after annotation using the BV-BRC annotation server. The sequences were annotated using the BV-BRC annotation server (https://www.bv-brc.org/) with default parameters. Protein annotations involved in virulence factors of the annotated genomes were downloaded using the specialty genes service of BV-BRC with the Virulence Factor Database (VFDB) filter, and the genes with their classification were used for subsequent analyses. For phylogenetic analysis, a total of 41 K. pneumoniae ST 307 including our isolates were used for SNP analysis. A completed genome of K. pneumoniae ST 307 strain Z0117KP0004 from a dog from South Korea (accession no. GCA_023657855.1) was used as a reference genome. High-quality SNPs analysis and maximum likelihood (ML) phylogenetic tree construction were conducted using the default quality filters in CSI phylogeny  and feces (KP-F) from the dog with bacteremia and enteritis in this study. The KP-B and KP-F isolates revealed the identical phenotypic and genotypic antibiotic resistance (aac (6′)-Ib-cr], aminoglycoside resistance genes , beta-lactam resistance genes , phenicol resistance gene (catB3), trimethoprim resistance gene (dfrA14), fosfomycin resistance gene (fosA), disinfectant resistance genes , quinolone resistance gene (qnrB1), sulphonamide resistance gene (sul2), and tetracycline resistance [tet (A)] ] . In addirA genes . The K. X − M−15 , 21. In X − M−15 , 23, blacteremia . Addition humans .K. pneumoniae ST 307 were available in BIGSdb-Pasteur from animals (n = 29), environment (n = 6), human (n = 299), and unknown source (n = 17). Additionally, 1,283 sequences were available in BV-BRC from animals (n = 31), environment n = 3), human , and unknown (n = 138). For phylogenetic analysis, 37 WGS of K. pneumoniae ST 307 from cats and dogs were downloaded from these databases. The phylogenetic tree of 41 genome sequences of the K. pneumoniae ST 307 is shown in Klebsiella spp. , magA (microviscosity-associated gene A), and genes encoding siderophores, such as aerobactin, enterobactin, and yersiniabactin , enterobactin , and yersiniabactin . All the other K. pneumoniae ST 307 isolates also harbored the virulence genes associated with adherence  and enterobactin , but 14 isolates of them carried the genes encoding yersiniabactin  using WGS. The isolates showed multidrug-resistance and harbored multiple antimicrobial resistance genes including blaCTX − M−15. The virulence gene profiles suggested that the K. pneumoniae ST 307 isolates were not hypervirulent K. pneumoniae but carried the genes encoding siderophores. This study is the first report on K. pneumoniae ST 307 from the bacteremia in a dog and the utilization of the WGS analysis to define the source of the bloodstream infection. It provides valuable reference data for genomic surveillance of new emerging K. pneumoniae ST 307 in companion animals alongside other well-known clones. Considering the emergence and rapid dissemination of high-risk multidrug-resistant K. pneumoniae in both companion animals and humans, surveillance strategies and genomic studies are essential in human and veterinary medicine.In this study, we report the WGS of https://www.ncbi.nlm.nih.gov/, PRJNA956693.The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found below: Sample collection for bacterial isolation involves procedures or treatments that fall under standard veterinary practices for diagnosing and treating animals, therefore, ethical approval was considered unnecessary. Written informed consent was obtained from the owners for the participation of their animals in this study.J-YH: Writing—original draft. Y-JC: Methodology, Writing—original draft. M-JJ: Methodology, Writing—original draft. D-HL: Supervision, Writing—review and editing. C-SS: Supervision, Writing—review and editing. J-HK: Conceptualization, Supervision, Writing—review and editing.


The cortex of adrenal gland produces glucocorticoid, mineralocorticoid, and androgen. The medulla of adrenal gland secrets catecholamines. These hormones play an important role in regulating blood pressure, metabolism, and homeostasis of glucose or electrolytes. Hypersecretion or hyposecretion by the adrenal gland will cause a complex cascade of hormone effects and lead to diseases, including Addison’s disease, Cushing’s syndrome, and congenital adrenal cortical hyperplasia. Skin is the largest organ of body. It provides protection and acts as a barrier against external damage factors like infectious organisms, chemicals, and allergens. Endocrinologic disorders often induce cutaneous abnormalities. According to the previous evidences, natural products have the potential properties for attenuating skin disorders and improving dermatologic symptoms by inhibiting inflammation through MAPK or PI3K/AKT-dependent NF-κB pathways. The natural products may also promote skin wound healing by inhibiting the production of matrix metalloproteinase-9. We systematically searched the relevant articles from databases, including PubMed, Embase, and Cochrane library databases, to review the effects of natural products on skin disorders. This article summarized the effects of natural products on skin inflammation caused by abnormal hormone secreted by adrenal gland. And the published papers indicated that natural products might be a potential source for treating skin diseases. Skin is a diverse and extensive organ of the human body. The functions of the skin are important for the healthy and aesthetic conditions . The skiAt present, the marketed drugs aim to reduce inflammation through regulating and/or suppressing immune responses and then improve function of skin, thus ameliorate clinical symptoms and signs with pruritus . It is wSapium glandulatum is a potential source of anti-inflammatory compounds for the treatment of the inflammation-derived skin diseases. The mechanism of action may be partially related to activating the glucocorticoid receptor. Research showed multiple treatments with Hydroalcoholic extract of S.apium glandulatum did not cause major adverse effects when evaluated in the skin atrophy model , and sex hormones. Adrenocorticotropin (ACTH) is secreted by the pituitary gland. ACTH binds to its receptor and promotes the conversion of cholesterol from cytoplasm to mitochondria into pregnenolone. Then, with the action of a series of enzymes, adrenocortical hormones are produced, which also become steroids hormone. Adrenal medulla secreted hormones including epinephrine, norepinephrine, and dopamine. The hormones secreted by adrenal glands are crucial for homeostasis of sodium, homeostasis of glucose, regulation of blood pressure and metabolism. Secretion of too much or not enough of the hormone will be life-threatening. Understanding the biological functions of adrenal hormones is a precondition to the treatment of adrenal gland disease .in vitro  system in human sebaceous cell in vitro . The mRNin vitro . Recent in vitro , androgein vitro , and estin vitro . Meanwhiin vitro . Furtherin vitro . Human sin vitro . It can in vitro .The endocrine system interacts with integumentary systems through a cohort of mechanisms. In the endocrinopathies, the dysregulation of endocrine hormones will lead to dermatologic disease . AbnormaGlucocorticoids are cardinal regulators of epidermal growth, differentiation, and homeostasis . GlucocoIn patients with low blood pressure, norepinephrine is commonly used to improve hemodynamic stability. It is reported that noradrenaline may result in peripheral gangrene . NoradreThe hormones secreted by the adrenal gland play a key role in maintaining body homeostasis. If adrenal gland secreted hormones too much or too little, which will cause skin inflammation leading to skin diseases.Recent evidence suggested that responses for endocrine stress not only are under control of the central nervous system but also occur in peripheral tissue . Skin isThe hormones secreted by adrenal gland play an important role in the metabolism , regulatDioscorea alata L., Smilax china L., and Trigonella foenum-graecum L.  is a traditional Chinese medicine . In Chinof mouse . It has ne level . The eff cortex. . In addi cortex.  signific cortex. . These rCurcuma longa L. (turmeric) (via inhibiting the release of inflammatory factors and keratinocyte proliferation (®) for improving signs of photoaging. The treatment showed a significant improvement in skin firmness and self-evaluations after 4 weeks usage (Curcumin  is a comurmeric) . Turmeriurmeric) . Previouurmeric) . Hormoneurmeric) , Addisonurmeric) . Hormoneurmeric) , skin aturmeric) , bruisinurmeric) , hyperpiurmeric) , hirsutiurmeric) , androgeurmeric) . Up to durmeric) , acne, surmeric) . It is rurmeric) . Curcumiferation . Curcumiferation . A cliniks usage . Clinicavia blocking PI3K/Akt signaling pathway, thereby attenuating NF-κB activation in skin inflammation. Glycyrrhizic acid can downregulate the expression of inducible nitric oxide synthase and cyclooxygenase-2 (2 18β-glycyrrhetinic acid cream or vehicle cream (without 18β-glycyrrhetinic acid) was applied twice daily for 7 consecutive days alleviates psoriasis-like diseases in mice and induces apoptosis of keratinocytes via PI3K/Akt signaling pathway (Glycyrrhizae Radix et Rhizoma is a native natural products to Asia and Mediterranean area . Glycyrrgenase-2 . Previougenase-2 . An intrgenase-2 . Patientgenase-2 . Clinicagenase-2 . Glycyrrgenase-2  in stomagenase-2 , which igenase-2 . 18β-glygenase-2 . It is r pathway . All pat pathway . In clinvia inhibiting IL-1β, IL-6, and TNF-α. Artemisinin also reduces the productions of chemokines of immune cells, such as CXCL2, CCL2, CCL20 and CXCL10 (+ T-cells (Artemisinin  is a sesd CXCL10 . Dihydrod CXCL10  is an acd CXCL10 . It has d CXCL10 . Recent d CXCL10 . Dihydrod CXCL10 . Studies T-cells , targeti T-cells , and reg T-cells . Furthervia Wnt/β-catenin signaling pathway, by inhibiting inflammation, or by enhancing the migration and proliferation of fibroblasts (Other natural products also alleviate skin inflammation induced by abnormal hormone secretion of adrenal gland through a range of mechanisms . For exaroblasts . Aloesinroblasts ., artemisinin, dihydroartemisinin are the main natural products to be evaluated during trials as a source of drug discovery for skin inflammation caused by abnormal secretion of hormones of adrenal gland.Disorder of hormone secretion by adrenal gland are closely associated with cutaneous lesions. Clinical presentation of skin atrophy, alopecia, psoriasis, skin pigmentation, rosacea, acne, atopic dermatitis, and hirsutism are frequently characterized by skin inflammation. The mechanism of skin inflammation is shown in


Pseudomonas aeruginosa is a Gram-negative opportunistic pathogen that causes high morbidity and mortality in cystic fibrosis (CF) and immunocompromised patients, including patients with ventilator-associated pneumonia (VAP), severely burned patients, and patients with surgical wounds. Due to the intrinsic and extrinsic antibiotic resistance mechanisms, the ability to produce several cell-associated and extracellular virulence factors, and the capacity to adapt to several environmental conditions, eradicating P. aeruginosa within infected patients is difficult. Pseudomonas aeruginosa is one of the six multi-drug-resistant pathogens (ESKAPE) considered by the World Health Organization (WHO) as an entire group for which the development of novel antibiotics is urgently needed. In the United States (US) and within the last several years, P. aeruginosa caused 27% of deaths and approximately USD 767 million annually in health-care costs. Several P. aeruginosa therapies, including new antimicrobial agents, derivatives of existing antibiotics, novel antimicrobial agents such as bacteriophages and their chelators, potential vaccines targeting specific virulence factors, and immunotherapies have been developed. Within the last 2–3 decades, the efficacy of these different treatments was tested in clinical and preclinical trials. Despite these trials, no P. aeruginosa treatment is currently approved or available. In this review, we examined several of these clinicals, specifically those designed to combat P. aeruginosa infections in CF patients, patients with P. aeruginosa VAP, and P. aeruginosa–infected burn patients. Pseudomonas aeruginosa is one of the most prevalent pathogens leading to acute and chronic infections in wounds, lung, and urinary tract ..P. aerugP. aeruginosa MEP antibodies were successfully produced in mice [P. aeruginosa infection in CF patients. The trial was concluded in 2000, but no results ensued (NCT00004747) [P. aeruginosa immune globulin (MEP IGIV) given over 1 year in reducing the frequency of acute pulmonary exacerbation in patients with cystic fibrosis, mild to moderate pulmonary disease, and mucoid P. aeruginosa colonization [P. aeruginosa colonies in the patient’s sputum, and improved the quality of life in patients infected with P. aeruginosa [P. aeruginosa immune globulin (MEP IVIG), high-dose MEP IVIG, or placebo [After opsonic  in mice , the imm in mice . These h0004747) . The clinization . The sturuginosa . A total placebo . The tre placebo .P. aeruginosa have also been pursued. Passive immunization using PcrV, a protein critical for the translocation of T3SS effectors, significantly enhanced the survival of mice in lung and burn models of P. aeruginosa infection [NCT00638365, NCT00691587, and NCT01695343) [NCT00691587) evaluated KB001 in patients in the intensive care setting who were receiving ventilator therapy and suffering from P. aeruginosa lung infections. Patients received either the placebo or one of two dose levels of KB001. KB001 (Humaneered™), which was used to treat P. aeruginosa infections, is a modified, PEGylated, recombinant anti-P. aeruginosa PcrV Fab’ antibody that binds with high affinity to the PcrV protein and blocks its activity [P. aeruginosa exotoxins into host immunological and epithelial cells depends on the protein PcrV, which is located close to the tip of the TTSS needle [P. aeruginosa infections [P. aeruginosa burden [Immunotherapeutic approaches targeting T3SS of nfection ,278. Subnfection ,280. Ant1695343) ,264,279.activity ,263. Thea burden . Patienta burden .P. aeruginosa infections (NCT00638365) [P. aeruginosa density, clinical results, and inflammatory markers [P. aeruginosa density, symptoms, or spirometry [P. aeruginosa TTSS in CF patients with persistent P. aeruginosa infection; it also decreased both airway inflammation and lung damage.A phase I/II clinical trial studied the safety, pharmacokinetic, and pharmacodynamic properties of KB001 in CF patients with chronic 0638365) . A singl0638365) . In each markers . KB001 hirometry . At day irometry . The cliP. aeruginosa infections (NCT01695343) [n = 83) or a placebo (n = 86) was given to a total of 169 patients [A phase I/II clinical trial also examined whether 16 weeks of KB001 treatment improved the time required for the use of antibiotics in patients with worsening respiratory symptoms. In addition, the trial examined KB001’s safety and efficacy in improving inflammatory markers and spirometry for patients with a history of chronic 1695343) . At leaspatients . Except patients . The timpatients . At weekpatients .P. aeruginosa [P. aeruginosa infections [P. aeruginosa strains expressing PsL [NCT02255760 and NCT02696902) [NCT02255760), healthy patients between the ages of 18 and 60 were given a single intravenous infusion of MEDI3902 to assess the drug’s safety, pharmacokinetics, antidrug antibody responses, ex vivo anticytotoxicity, and opsonophagocytic killing capabilities against P. aeruginosa. Using randomization, 56 patients were randomly assigned to receive either 250, 750, 1500, or 3000 mg of MEDI3902 or a placebo over a period of 60 days [P. aeruginosa pneumonia.The Psl exopolysaccharide is another virulence factor present in numerous strains of ruginosa . Antibodfections . The Pslsing PsL . It was sing PsL . A bispe2696902) ,266,283. 60 days . There w 60 days . The resP. aeruginosa (NCT02696902) [P. aeruginosa colonization of the lower respiratory tract who were randomized into either a single IV infusion of 1500 mg MEDI3902 (n = 85) or placebo (n = 83) [P. aeruginosa pneumonia as the primary efficacy outcome.A phase II clinical trial examined the efficacy, pharmacokinetics, and safety of MEDI3902 in mechanically ventilated ICU patients infected with 2696902) . The stu(n = 83) . The stuP. aeruginosa vaccination were used to create “anti-pseudomonas IgY”, called PseudIgY (avian Ab to P. aeruginosa). The trial assessed whether daily gargling with PsAer-IgY against P. aeruginosa prevents P. aeruginosa infections in CF patients (NCT00633191) [P. aeruginosa infections. In the trial, patients received a brief course of antibiotics to eliminate P. aeruginosa infection. After that, they began gargling with a PseudIgY solution every night to ward off infection [In another study, eggs from chickens that have received a 0633191) . The stunfection .P. aeruginosa following a successful treatment of acute or intermittent infection (NCT01455675) [Another phase III double-blind placebo-controlled trial investigated whether PseudIgY prolongs the time to reinfection with 1455675) . The Pse1455675) . In the 1455675) .P. aeruginosa. One of the targeted antigens was the O-polysaccharide moiety of P. aeruginosa serotype O11. A human monoclonal IgM antibody, KBPA-101 or panobacumab, was developed and found to be safe and efficacious in preclinical in vitro and in vivo studies and in phase I clinical trials (NCT00851435) [P. aeruginosa O11 pneumonia [Several immunotherapeutic trials have been conducted targeting different antigens of 0851435) ,270,271.0851435) ,270,271.0851435) ,270,271.neumonia ,270,271.P. aeruginosa alginate, AR-105 (Aerucin®), as adjunctive therapeutic treatment to standard-of-care antibiotics for P. aeruginosa pneumonia in mechanically ventilated patients (NCT03027609) [More recently, a phase II clinical trial was conducted to assess the efficacy, safety, and pharmacokinetics of a new human monoclonal antibody against 3027609) . The stu3027609) . The 158E. coli to cause sepsis in two animal models: Syrian golden hamsters [®) attenuated the endotoxin (LPS) activity by reducing inflammation. A clinical trial examined the efficacy of IgM-enriched intravenous immunoglobulin (Pentaglobin®—5 mL/kg over a 12 h intravenous infusion for 3 consecutive days) to decrease mortality in 120 neutropenic acute leukemia or hematopoietic stem cell transplant patients colonized with carbapenem-resistant Enterobacteriaceae or P. aeruginosa (NCT03494959) [Recent studies were conducted to elucidate the mechanism of action of IgM-enriched immunoglobulin (IgM-IVIg) . The stuhamsters  or the Chamsters . Furtherhamsters . All thr3494959) .P. aeruginosa species [P. aeruginosa, is a catastrophic consequence that leads to prolonged hospitalization and higher medical costs [P. aeruginosa is linked to osteomyelitis, septic arthritis, and prosthetic joint infections due to its ability to adhere to bone and fibrocartilaginous articular structures. A phase I randomized clinical trial (NCT04763759) was designed to assess the overall safety and pharmacokinetics of TRL1068 in 18 patients with prosthetic joints using three separate dosages: 6, 15, and 30 mg/kg [P. aeruginosa and the level of C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), IL-6, and IL-10 [Another promising novel treatment, TRL1068, a human monoclonal antibody that targets DNA-binding protein II (DNABII), is currently undergoing clinical testing. DNABII is an important structural element of biofilms because it stabilizes extracellular DNA ,289,290. species ,291.Thoual costs ,293,294.30 mg/kg . The stund IL-10 .P. aeruginosa hybrid protein, OprF/I  involved active immunization with a , OprF/I . In the ruginosa ,296. In m models ,296.NCT00778388) [A phase I trial examined the immunogenicity, safety, and tolerance in a healthy adult immunized with three different dosages of IC43 compared with placebo (0778388) . Two int0778388) . In heal0778388) .NCT00876252) [P. aeruginosa from the blood, wounds, respiratory tract, urine, and central venous catheter [P. aeruginosa diagnosis between each visit up to day 90 [Pseudomonas aeruginosa infection rates were not significantly different, and the IC43 groups had a low incidence of invasive infections  [P. aeruginosa infection rates did not differ significantly among the four treatment groups [A subsequent phase II study in which IC43 was used to immunize mechanically ventilated ICU patients has been completed using doses of 100 and 200 μg with adjuvant and a dose of 100 μg without adjuvant (0876252) . Patient0876252) . At visicatheter . When meo day 90 . All IC4o day 90 . Pseudomeumonia) . In the eumonia) . No fataeumonia) . Overallt groups .P. aeruginosa vaccine in nonsurgical ICU patients (NCT01563263) [P. aeruginosa invasive infection or respiratory tract infection were similar [Another double-blind phase II/III clinical trial examined the efficacy, immunogenicity, and safety of IC43 recombinant 1563263) . Eight h1563263) . All-cau1563263) . At day 1563263) . Both gr similar . Compare similar . Respira similar . In the  similar . The IC4 similar . High im similar .P. aeruginosa infections have been investigated, including inhaled gallium and sodium nitrite  in P. aeruginosa infected CF (NCT01093521) [P. aeruginosa respiratory infections [®) infusion for 5 days with either 100 or 200 mg/m2.In addition to the previous treatments, other alternatives for eliminating  nitrite  [157,300P. aeruginosa was conducted to evaluate the efficacy of IV gallium in improving pulmonary function as measured by a 5% or greater relative improvement in forced expiratory volume in 1 s (FEV1) from baseline to day 28 (NCT02354859) [2/day) or normal saline over 5 days [A subsequent phase II, multicenter, randomized, placebo-controlled trial in adult CF patients chronically infected with 2354859) . The 119r 5 days .Pseudomonas infections to reduce the burden of P. aeruginosa NCT02694393) [P. aeruginosa within the lung of CF-infected patients by increasing the susceptibility of P. aeruginosa to antibiotics [In addition to gallium, another phase I/II study assessed the safety of inhaled sodium nitrite in 35 adults with CF and chronic 2694393) . Sodium ibiotics ,305. PatNCT03032380) was designed to compare all-cause mortality at day 14 in patients receiving cefiderocol or meropenem for HABP, VABP, or health-care-associated bacterial pneumonia (HCABP) caused by Gram-negative pathogens [n = 148) or meropenem (n = 152) every 8 h. Moreover, open-label intravenous linezolid was administered to all patients for a minimum of 5 days [Cefedrolor is a novel siderophore cephalosporin with broad activity and high potency against multi-drug-resistant Gram-negative bacteria ,306. A pP. aeruginosa vaccine or therapy is long and challenging. Antibiotics will still be essential in combating P. aeruginosa infections. For example, other clinical trials have examined newer combinations of beta-lactamases, polymyxins, and alternative formulations, such as liposomal or inhalation, for traditional antibiotics against P. aeruginosa infections [P. aeruginosa mutants. Therefore, developing and testing several nontraditional therapies is critical. These therapies may not be alternatives to or replace the current antibiotics. Rather, they may be used in conjunction with antibiotics to help reduce the dose of antibiotics required to treat infected patients, which will lessen the production of P. aeruginosa bioburden and/or reduce the rate of emergence of antibiotic resistant mutants.It is evident from the outcomes of the different clinical trials described in this review that the road to developing an effective anti-fections ,320,321.P. aeruginosa treatments described in this review include: bacteriophage; vaccines targeting outer membrane proteins (opRF/opRI); and treatment targeting an essential virulence factor, such as the QS system, T3SS, and monoclonal antibodies. As we show in this review, many of these approaches produced encouraging results. In addition, and except for bacteriophages, these treatments are less likely to exert pressure on P. aeruginosa to develop resistant mutants. Bacteriophage therapy shares with antibiotics the inherit problem of emerging phage- or antibiotic-resistant mutants that may result from the overuse of these bacteriophages. However, unlike antibiotics, the application of multiple phages (cocktails) will significantly reduce the problem associated with the emergence of antibiotic resistant mutants.Besides antibiotics, anti-P. aeruginosa virulence factor, such as outer membrane proteins or a system/mechanism that could help the production of multiple virulence factors, such as the QS system, T3SS, and iron-acquisition system. However, a key factor that needs to be considered in developing these therapies is the pathogenesis of P. aeruginosa in different acute and chronic infections, including acute pneumonia, VAP, burn wound infections, urinary tract infections, and chronic lung infections in CF patients. Several previous studies have already demonstrated that the capacity of P. aeruginosa to cause different infections lies in its ability to tailor the production of different cell-associated and extracellular virulence factors in response to environmental cues within certain infection sites, including lung wounds, blood, or urinary tract. Therefore, to further develop current therapies and to identify new targets for further therapies, it is critical to define a specific virulent factor(s) that is produced in response to the in vivo environment. Previous studies analyzed P. aeruginosa pathogenesis utilizing growth conditions that mimic the in vivo conditions. For example, Palmer et al. [P. aeruginosa during chronic lung infections of CF patients, while Kruczek utilized whole blood to examine the pathogenesis of P. aeruginosa infections in severely burned patients [P. aeruginosa in CF sputum differentially expressed the iron-acquisition genes and flagella motility genes and enhanced the production of the cell-to-cell communication molecule, PQS [P. aeruginosa iron-acquisition system. Additional anti-P. aeruginosa therapies to treat lung infections in CF patients may target the synthesis and/or the secretion of the PQS molecule. Similarly, Kruczek et al. showed that compared with the growth in blood from healthy volunteers, the growth of P. aeruginosa in blood from severely burned patients significantly increased the expression of more than 1000 genes, including genes encoding the QS-controlled virulence factors and those encoding the transport heme and phosphate. Any of these systems is a likely target for further therapy to prevent bacteremia and sepsis in P. aeruginosa–infected severely burned patients.Among the different nontraditional therapies described in this review, the most promising one is targeting a specific r et al.  utilizedpatients . Palmer ule, PQS . In addiule, PQS . As we sP. aeruginosa. The PcrV protein plays a pivotal role in the pathogenesis of P. aeruginosa infections. The production of several cell-associated and extracellular virulence factors strongly indicates the reliance of P. aeruginosa on many of these factors in establishing infection at different sites within the host. However, despite these factors, previous studies using the murine model of lung infection and the murine model of thermal injury clearly showed that specific PcrV antibodies only compromised the ability of P. aeruginosa to cause lung infection and bacteremia/sepsis, respectively [P. aeruginosa lung infections in CF patients was assessed using the clinical trials described in P. aeruginosa bacterial load at the infection site [P. aeruginosa–induced bacteremia/sepsis by affecting the translocation of P. aeruginosa into the bloodstream. Therefore, it is possible that future clinical studies may prove that the PcrV monoclonal antibodies are more effective in preventing P. aeruginosa bacteremia in severely burned patients than effectively treating P. aeruginosa lung infections in CF patients. In addition, future studies may identify a potential target, similar to PcrV, that plays a critical role in the pathogenesis of P. aeruginosa infections. A treatment containing a combination of antibodies to PcrV and such a target may prove to be more effective than PcrV antibody alone in treating lung infections in CF patients. The antibodies may also be utilized in synergy with currently available antibiotics in treating P. aeruginosa lung infections.As we demonstrated in this review, several clinical trials were focused on assessing the effectiveness of new antibiotics targeting the PcrV protein . The 32.ectively ,323. Basion site . This su