O26 Polysaccharides as Key Players in Enteropathogenic E. coli Immune Evasion and Vaccine Development.

Enteropathogenic Escherichia coli (EPEC) produce a capsule of polysaccharides identical to those composing the O-antigen polysaccharide of its LPS (lipopolysaccharide) molecules. In light of this, the impact of O26 polysaccharides on the immune evasion mechanisms of capsulated O26 EPEC compared to non-capsulated enterohemorrhagic Escherichia coli (EHEC) was investigated. Our findings reveal that there was no significant difference between the levels in EPEC and EHEC of rhamnose (2.8:2.5), a molecule considered to be a PAMP (Pathogen Associated Molecular Patterns). However, the levels of glucose (10:1.69), heptose (3.6:0.89) and N-acetylglucosamine (4.5:2.10), were significantly higher in EPEC than EHEC, respectively. It was also observed that the presence of a capsule in EPEC inhibited the deposition of C3b on the bacterial surface and protected the pathogen against lysis by the complement system. In addition, the presence of a capsule also protected EPEC against phagocytosis by macrophages. However, the immune evasion provided by the capsule was overcome in the presence of anti-O26 polysaccharide antibodies, and additionally, these antibodies were able to inhibit O26 EPEC adhesion to human epithelial cells. Finally, the results indicate that O26 polysaccharides can generate an effective humoral immune response, making them promising antigens for the development of a vaccine against capsulated O26 E. coli.

O26 Polysaccharides as key players in enteropathogenic E. coli immune evasion and vaccine development

Enteropathogenic Escherichia coli (EPEC) produce a capsule of polysaccharides identical to those composing the O-antigen polysaccharide of its LPS (lipopolysaccharide) molecules. In light of this, the impact of O26 polysaccharides on the immune evasion mechanisms of capsulated O26 EPEC compared to non-capsulated enterohemorrhagic Escherichia coli (EHEC) was investigated. Our findings reveal that there was no significant difference between the levels in EPEC and EHEC of rhamnose (2.8:2.5), a molecule considered to be a PAMP (Pathogen Associated Molecular Patterns). However, the levels of glucose (10:1.69), heptose (3.6:0.89) and N-acetylglucosamine (4.5:2.10), were significantly higher in EPEC than EHEC, respectively. It was also observed that the presence of a capsule in EPEC inhibited the deposition of C3b on the bacterial surface and protected the pathogen against lysis by the complement system. In addition, the presence of a capsule also protected EPEC against phagocytosis by macrophages. However, the immune evasion provided by the capsule was overcome in the presence of anti-O26 polysaccharide antibodies, and additionally, these antibodies were able to inhibit O26 EPEC adhesion to human epithelial cells. Finally, the results indicate that O26 polysaccharides can generate an effective humoral immune response, making them promising antigens for the development of a vaccine against capsulated O26 E. coli.

Synthesis, characterization, antimicrobial activity, and toxicity evaluation of aminolevulinic acid-silver and silver-iron nanoparticles for potential applications in agriculture.

Aminolevulinic acid (ALA) is considered one of the most critical plants growth regulators and essential precursors for chlorophyll biosynthesis; besides, its photodynamic activity can be used to exterminate larvae and microorganisms in plants and soil. Silver nanoparticles (AgNPs) have unique physicochemical properties and potent antimicrobial, antiviral, and antifungal activities, and in agriculture, their application as nanopesticides has been proposed. In this study, silver and silver-iron nanoparticles capped/stabilized with aminolevulinic acid (ALAAgNPs and ALAAgFeNPs) were synthesized by the photoreduction method and characterized by UV-vis spectroscopy, transmission electron microscopy, and zeta potential analysis. The kinetics of 1O2 generation from ALAAgFeNPs were obtained. The ALANP toxicity was evaluated on stalks of E. densa by observing cell morphology changes and measuring chlorophyll content compared with water-treated plants. Antimicrobial activity was tested against E. coli, P. aeruginosa, and Candida albicans. The results suggested that ALANPs (prepared with [AgNO3] ≤ 0.2 mM and [ALA] ≤ 0.4 mM) could be suitable for applications in the agricultural sector. The presence of ∼0.3 mmol of iron in ALAAgNPs synthesis increased cell uptake and chlorophyll synthesis.

Antimicrobial Activity of Silver and Gold Nanoparticles Prepared by Photoreduction Process with Leaves and Fruit Extracts of Plinia cauliflora and Punica granatum.

The increased number of resistant microbes generates a search for new antibiotic methods. Metallic nanoparticles have emerged as a new platform against several microorganisms. The nanoparticles can damage the bacteria membrane and DNA by oxidative stress. The photoreduction process is a clean and low-cost method for obtaining silver and gold nanoparticles. This work describes two original insights: (1) the use of extracts of leaves and fruits from a Brazilian plant Plinia cauliflora, compared with a well know plant Punica granatum, and (2) the use of phytochemicals as stabilizing agents in the photoreduction process. The prepared nanoparticles were characterized by UV-vis, FTIR, transmission electron microscopy, and Zeta potential. The antimicrobial activity of nanoparticles was obtained with Gram-negative and Gram-positive bacteria, particularly the pathogens Staphylococcus aureus ATCC 25923; Bacillus subtilis ATCC 6633; clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA) and Enterococcus faecalis; Escherichia coli ATCC 25922; Escherichia coli O44:H18 EAEC042 (clinical isolate); Klebsiella pneumoniae ATCC 700603, Salmonella Thiphymurium ATCC 10231; Pseudomonas aeruginosa ATCC 27853; and Candida albicans ATCC 10231. Excellent synthesis results were obtained. The AgNPs exhibited antimicrobial activities against Gram-negative and Gram-positive bacteria and yeast (80-100%), better than AuNPs (0-87.92%), and may have the potential to be used as antimicrobial agents.

O55 Polysaccharides Are Good Antigen Targets for the Formulation of Vaccines against O55 STEC and Capsulated aEPEC Strains.

The serogroup O55 of E. coli is composed of strains whose mechanisms of virulence are different from each other. Since the O55 polysaccharides are present in all E. coli O55 strains, and so are the polymers that compose the capsule of O55 atypical enteropathogenic E. coli (aEPEC), it was investigated whether anti-O55 antibodies were able to help the innate immune system to eliminate capsulated aEPEC and Shiga toxin-producing E. coli (STEC) belonging to the serogroup O55. The results demonstrate that the capsule of EPEC was able to inhibit the deposition of C3b on the bacterial surface and, as a consequence, their lysis by the alternative pathway of the complement system. However, in the presence of antibodies, the ability of the complement to lyse these pathogens was restored. It was also observed that macrophages were able to ingest EPEC and STEC, but they were only able to kill the ingested pathogens in the presence of antibodies. Anti-O55 antibodies were also able to inhibit aEPEC and STEC O55 adherence to human epithelial cells. In summary, the results demonstrated that the O55 polysaccharides have the potential to induce an effective humoral immune response against STEC and EPEC, indicating that they are good antigen targets to be used in vaccine formulations against these pathogens.

Eugenia uniflora L. Silver and Gold Nanoparticle Synthesis, Characterization, and Evaluation of the Photoreduction Process in Antimicrobial Activities.

Eugenia uniflora linnaeus, known as Brazilian cherry, is widely distributed in Brazil, Argentina, Uruguay, and Paraguay. E. uniflora L. extracts contain phenolic compounds, such as flavonoids, tannins, triterpenes, and sesquiterpenes. The antimicrobial action of essential oils has been attributed to their compositions of bioactive compounds, such as sesquiterpenes. In this paper, the fruit extract of E. uniflora was used to synthesize silver and gold nanoparticles. The nanoparticles were characterized by UV-Vis, transmission electron microscopy, elemental analysis, FTIR, and Zeta potential measurement. The silver and gold nanoparticles prepared with fruit extracts presented sizes of ~32 nm and 11 nm (diameter), respectively, and Zeta potentials of -22 mV and -14 mV. The antimicrobial tests were performed with Gram-negative and Gram-positive bacteria and Candida albicans. The growth inhibition of EuAgNPs prepared with and without photoreduction showed the important functional groups in the antimicrobial activity.

Escherichia coli Strains Responsible for Cystitis in Female Pediatric Patients with Normal and Abnormal Urinary Tracts Have Different Virulence Profiles.

The role of uropathogenic Escherichia coli (UPEC) in colonization and infection of female patients with anatomical and functional abnormalities of the urinary system is elusive. In this study, the phenotype, genotype and the phylogeny of UPEC strains isolated from the urine of pediatric female patients with cystitis of normal and abnormal urinary tract were determined. Multiplex PCR results demonstrated that 86% of the strains isolated from female patients with normal urinary tract (NUT), belonged to the phylo-groups B2 and D. Their prevalence decreased to 23% in strains isolated from patients with abnormal urinary tract (AUT). More of the isolates from AUT patients produced a biofilm on polystyrene and polyvinyl chloride (PVC), adhered to epithelial cells, and encoded pap and sfa genes than strains isolated from female patients with NUT. In contrast, a higher number of hemolysin-producing strains with serogroups associated with UPEC were isolated from patients with NUT. In summary, the results suggest that cystitis in female patients with NUT is associated with ExPEC, whereas cystitis in female patients with AUT is associated with pathogenic intestinal E. coli strains that have acquired the ability to colonize the bladder.

Antimicrobial activity of silver and gold nanoparticles prepared by photoreduction process with leaves and fruit extracts of Plinia cauliflora and Punica granatum

The increased number of resistant microbes generates a search for new antibiotic methods. Metallic nanoparticles have emerged as a new platform against several microorganisms. The nanoparticles can damage the bacteria membrane and DNA by oxidative stress. The photoreduction process is a clean and low-cost method for obtaining silver and gold nanoparticles. This work describes two original insights: (1) the use of extracts of leaves and fruits from a Brazilian plant Plinia cauliflora, compared with a well know plant Punica granatum, and (2) the use of phytochemicals as stabilizing agents in the photoreduction process. The prepared nanoparticles were characterized by UV-vis, FTIR, transmission electron microscopy, and Zeta potential. The antimicrobial activity of nanoparticles was obtained with Gram-negative and Gram-positive bacteria, particularly the pathogens Staphylococcus aureus ATCC 25923; Bacillus subtilis ATCC 6633; clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA) and Enterococcus faecalis; Escherichia coli ATCC 25922; Escherichia coli O44:H18 EAEC042 (clinical isolate); Klebsiella pneumoniae ATCC 700603, Salmonella Thiphymurium ATCC 10231; seudomonas aeruginosa ATCC 27853; and Candida albicans ATCC 10231. Excellent synthesis results were obtained. The AgNPs exhibited antimicrobial activities against Gram-negative and Gram-positive bacteria and yeast (80–100%), better than AuNPs (0–87.92%), and may have the potential to be used as antimicrobial agents.

Synthesis, characterization, antimicrobial activity, and toxicity evaluation of aminolevulinic acid–silver and silver–iron nanoparticles for potential applications in agriculture

Aminolevulinic acid (ALA) is considered one of the most critical plants growth regulators and essential precursors for chlorophyll biosynthesis; besides, its photodynamic activity can be used to exterminate larvae and microorganisms in plants and soil. Silver nanoparticles (AgNPs) have unique physicochemical properties and potent antimicrobial, antiviral, and antifungal activities, and in agriculture, their application as nanopesticides has been proposed. In this study, silver and silver–iron nanoparticles capped/stabilized with aminolevulinic acid (ALAAgNPs and ALAAgFeNPs) were synthesized by the photoreduction method and characterized by UV-vis spectroscopy, transmission electron microscopy, and zeta potential analysis. The kinetics of 1O2 generation from ALAAgFeNPs were obtained. The ALANP toxicity was evaluated on stalks of E. densa by observing cell morphology changes and measuring chlorophyll content compared with water-treated plants. Antimicrobial activity was tested against E. coli, P. aeruginosa, and Candida albicans. The results suggested that ALANPs (prepared with [AgNO3] ≤ 0.2 mM and [ALA] ≤ 0.4 mM) could be suitable for applications in the agricultural sector. The presence of ∼0.3 mmol of iron in ALAAgNPs synthesis increased cell uptake and chlorophyll synthesis.

O55 polysaccharides are good antigen targets for the formulation of vaccines against O55 STEC and capsulated aEPEC strains

The serogroup O55 of E. coli is composed of strains whose mechanisms of virulence are different from each other. Since the O55 polysaccharides are present in all E. coli O55 strains, and so are the polymers that compose the capsule of O55 atypical enteropathogenic E. coli (aEPEC), it was investigated whether anti-O55 antibodies were able to help the innate immune system to eliminate capsulated aEPEC and Shiga toxin-producing E. coli (STEC) belonging to the serogroup O55. The results demonstrate that the capsule of EPEC was able to inhibit the deposition of C3b on the bacterial surface and, as a consequence, their lysis by the alternative pathway of the complement system. However, in the presence of antibodies, the ability of the complement to lyse these pathogens was restored. It was also observed that macrophages were able to ingest EPEC and STEC, but they were only able to kill the ingested pathogens in the presence of antibodies. Anti-O55 antibodies were also able to inhibit aEPEC and STEC O55 adherence to human epithelial cells. In summary, the results demonstrated that the O55 polysaccharides have the potential to induce an effective humoral immune response against STEC and EPEC, indicating that they are good antigen targets to be used in vaccine formulations against these pathogens.

Eugenia uniflora L. silver and gold nanoparticle synthesis, characterization, and evaluation of the photoreduction process in antimicrobial activities

Eugenia uniflora linnaeus, known as Brazilian cherry, is widely distributed in Brazil, Argentina, Uruguay, and Paraguay. E. uniflora L. extracts contain phenolic compounds, such as flavonoids, tannins, triterpenes, and sesquiterpenes. The antimicrobial action of essential oils has been attributed to their compositions of bioactive compounds, such as sesquiterpenes. In this paper, the fruit extract of E. uniflora was used to synthesize silver and gold nanoparticles. The nanoparticles were characterized by UV–Vis, transmission electron microscopy, elemental analysis, FTIR, and Zeta potential measurement. The silver and gold nanoparticles prepared with fruit extracts presented sizes of ~32 nm and 11 nm (diameter), respectively, and Zeta potentials of −22 mV and −14 mV. The antimicrobial tests were performed with Gram-negative and Gram-positive bacteria and Candida albicans. The growth inhibition of EuAgNPs prepared with and without photoreduction showed the important functional groups in the antimicrobial activity.

Escherichia coli strains responsible for cystitis in female pediatric patients with normal and abnormal urinary tracts have different virulence profiles

The role of uropathogenic Escherichia coli (UPEC) in colonization and infection of female patients with anatomical and functional abnormalities of the urinary system is elusive. In this study, the phenotype, genotype and the phylogeny of UPEC strains isolated from the urine of pediatric female patients with cystitis of normal and abnormal urinary tract were determined. Multiplex PCR results demonstrated that 86% of the strains isolated from female patients with normal urinary tract (NUT), belonged to the phylo-groups B2 and D. Their prevalence decreased to 23% in strains isolated from patients with abnormal urinary tract (AUT). More of the isolates from AUT patients produced a biofilm on polystyrene and polyvinyl chloride (PVC), adhered to epithelial cells, and encoded pap and sfa genes than strains isolated from female patients with NUT. In contrast, a higher number of hemolysin-producing strains with serogroups associated with UPEC were isolated from patients with NUT. In summary, the results suggest that cystitis in female patients with NUT is associated with ExPEC, whereas cystitis in female patients with AUT is associated with pathogenic intestinal E. coli strains that have acquired the ability to colonize the bladder.

Inactivation of the antimicrobial peptide LL-37 by pathogenic Leptospira

Leptospires are aerobic, Gram-negative spirochetes with a high invasive capacity. Pathogenic leptospires secrete proteases that inactivate a variety of host's proteins including molecules of the extracellular matrix and of the human complement system. This strategy, used by several pathogens of medical importance, contributes to bacterial invasion and immune evasion. In the current work we present evidence that Leptospira proteases also target human cathelicidin (LL-37), an antimicrobial peptide that plays an important role in the innate immune response. By using six Leptospira strains, four pathogenic and two saprophytic, we demonstrated that proteases present in the supernatants of pathogenic strains were capable of degrading LL-37 in a time-dependent manner, whereas proteolytic degradation was not observed with the supernatants of the two saprophytic strains. Inactivation of LL-37 was prevented by using the 1,10-phenanthroline inhibitor, thus suggesting the involvement of metalloproteinases in this process. In addition, the antibacterial activity of LL-37 against two Leptospira strains was evaluated. Compared to the saprophytic strain, a greater resistance of the pathogenic strain to the action of the peptide was observed. Our data suggest that the capacity to inactivate the host defense peptide LL-37 may be part of the virulence arsenal of pathogenic Leptospira, and we hypothesize that its inactivation by the bacteria may influence the outcome of the disease.

A new class of antimicrobial molecules derived from kefir, effective against Pseudomonas aeruginosa and methicillin resistant Staphylococcus aureus (MRSA) strains.

Many studies have linked the antimicrobial properties of kefir with the presence of bacteriocins and organic acids. In the present work, results obtained from bacteriostatic and bactericidal studies, and from RP-HPLC, Mass Spectrometry and proton NMR analysis, show that a sample of milk kefir grains is able to produce an antimicrobial fraction, denoted FK-1000, composed of sugars and amino acids, predominantly polymers of alanine, doublets of tyrosine and phenylalanine. Since this fraction is a lyophilized product whose molecular profile is different from bacteriocins and simple carboxylic acids, its antimicrobial effect cannot be attributed to these molecules, or to alcohols or hydrogen peroxide. The fraction is bactericidal against weak-acid-resistant MRSA and weak-acid resistant P. aeruginosa at pH 5, and is bacteriostatic against both pathogens at pH 7. In combination formulation, the FK-1000 fraction is able to increase fivefold the effect of streptomycin against P. aeruginosa and it is not toxic to human epithelial cells at antimicrobial concentrations. 16 S rRNA microbiota analysis of antimicrobial-producing and non-producing kefir grains demonstrated that they are distinct. In summary, the results indicate that milk kefir grains can produce different classes of molecules with potent antibiotic activity against resistant bacteria.

Inactivation of the antimicrobial peptide LL-37 by pathogenic Leptospira

Leptospires are aerobic, Gram-negative spirochetes with a high invasive capacity. Pathogenic leptospires secrete proteases that inactivate a variety of host's proteins including molecules of the extracellular matrix and of the human complement system. This strategy, used by several pathogens of medical importance, contributes to bacterial invasion and immune evasion. In the current work we present evidence that Leptospira proteases also target human cathelicidin (LL-37), an antimicrobial peptide that plays an important role in the innate immune response. By using six Leptospira strains, four pathogenic and two saprophytic, we demonstrated that proteases present in the supernatants of pathogenic strains were capable of degrading LL-37 in a time-dependent manner, whereas proteolytic degradation was not observed with the supernatants of the two saprophytic strains. Inactivation of LL-37 was prevented by using the 1,10-phenanthroline inhibitor, thus suggesting the involvement of metalloproteinases in this process. In addition, the antibacterial activity of LL-37 against two Leptospira strains was evaluated. Compared to the saprophytic strain, a greater resistance of the pathogenic strain to the action of the peptide was observed. Our data suggest that the capacity to inactivate the host defense peptide LL-37 may be part of the virulence arsenal of pathogenic Leptospira, and we hypothesize that its inactivation by the bacteria may influence the outcome of the disease.

A new class of antimicrobial molecules derived from kefir, effective against Pseudomonas aeruginosa and methicillin resistant Staphylococcus aureus (MRSA) strains

Many studies have linked the antimicrobial properties of kefir with the presence of bacteriocins and organic acids. In the present work, results obtained from bacteriostatic and bactericidal studies, and from RP-HPLC, Mass Spectrometry and proton NMR analysis, show that a sample of milk kefir grains is able to produce an antimicrobial fraction, denoted FK-1000, composed of sugars and amino acids, predominantly polymers of alanine, doublets of tyrosine and phenylalanine. Since this fraction is a lyophilized product whose molecular profile is different from bacteriocins and simple carboxylic acids, its antimicrobial effect cannot be attributed to these molecules, or to alcohols or hydrogen peroxide. The fraction is bactericidal against weak-acid-resistant MRSA and weak-acid resistant P. aeruginosa at pH 5, and is bacteriostatic against both pathogens at pH 7. In combination formulation, the FK-1000 fraction is able to increase fivefold the effect of streptomycin against P. aeruginosa and it is not toxic to human epithelial cells at antimicrobial concentrations. 16 S rRNA microbiota analysis of antimicrobial-producing and non-producing kefir grains demonstrated that they are distinct. In summary, the results indicate that milk kefir grains can produce different classes of molecules with potent antibiotic activity against resistant bacteria.

Tryptophan Silver Nanoparticles Synthesized by Photoreduction Method: Characterization and Determination of Bactericidal and Anti-Biofilm Activities on Resistant and Susceptible Bacteria.

The high rates of antibiotics use in hospitals have resulted in a condition where multidrug-resistant pathogens have become a severe threat to the human health worldwide. Therefore, there is an increasing necessity to identify new antimicrobial agents that can inhibit the multidrug-resistant bacteria and biofilm formation. In this study, antibacterial and anti-biofilm activities of tryptophan silver nanoparticles (TrpAgNP) were investigated. The TrpAgNPs were synthesized by photoreduction method, and the influence of irradiation time and concentration of reagents were analyzed. The nanoparticles were characterized by transmission electron microscopy, Zeta Potential and (UV)-absorption spectra. The antibacterial activity of TrpAgNPs was tested for antibiotic-resistant and susceptible pathogens, Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Citrobacter freundii, Klebsiella pneumoniae, Salmonella typhimurium, and Pseudomonas aeruginosa, evaluating the influence of photoreduction parameters in bactericidal effect. The results have shown that TrpAgNPs solutions with lower tryptophan/silver nitrate (AgNO3) ratio and higher AgNO3 concentration have higher bactericidal action against bacteria with inhibition of ~100% in almost all studied bacterial strains. The antimicrobial activity of TrpAgNPs within biofilms generated under static conditions of antibiotic-resistant and susceptible strains of S. aureus, S. epidermidis, E. coli, K. pneumoniae, C. freundii, and P. aeruginosa was also investigated. The results showed that TrpAgNPs have an inhibitory effect against biofilm formation, exceeding 50% in the case of Gram-negative bacteria (E. coli, K. pneumoniae, C. freundii, and P. aeruginosa-54.8% to 98.8%). For Gram-positive species, an inhibition of biofilm formation of 68.7% to 72.2 % was observed for S. aureus and 20.0% to 40.2% for S. epidermidis.

Tryptophan silver nanoparticles synthesized by photoreduction method: characterization and determination of bactericidal and anti-biofilm activities on resistant and susceptible bacteria

The high rates of antibiotics use in hospitals have resulted in a condition where multidrug-resistant pathogens have become a severe threat to the human health worldwide. Therefore, there is an increasing necessity to identify new antimicrobial agents that can inhibit the multidrug-resistant bacteria and biofilm formation. In this study, antibacterial and anti-biofilm activities of tryptophan silver nanoparticles (TrpAgNP) were investigated. The TrpAgNPs were synthesized by photoreduction method, and the influence of irradiation time and concentration of reagents were analyzed. The nanoparticles were characterized by transmission electron microscopy, Zeta Potential and (UV)-absorption spectra. The antibacterial activity of TrpAgNPs was tested for antibiotic-resistant and susceptible pathogens, Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Citrobacter freundii, Klebsiella pneumoniae, Salmonella typhimurium, and Pseudomonas aeruginosa, evaluating the influence of photoreduction parameters in bactericidal effect. The results have shown that TrpAgNPs solutions with lower tryptophan/silver nitrate (AgNO3) ratio and higher AgNO3 concentration have higher bactericidal action against bacteria with inhibition of ~100% in almost all studied bacterial strains. The antimicrobial activity of TrpAgNPs within biofilms generated under static conditions of antibiotic-resistant and susceptible strains of S. aureus, S. epidermidis, E. coli, K. pneumoniae, C. freundii, and P. aeruginosa was also investigated. The results showed that TrpAgNPs have an inhibitory effect against biofilm formation, exceeding 50% in the case of Gram-negative bacteria (E. coli, K. pneumoniae, C. freundii, and P. aeruginosa—54.8% to 98.8%). For Gram-positive species, an inhibition of biofilm formation of 68.7% to 72.2 % was observed for S. aureus and 20.0% to 40.2% for S. epidermidis.KeywoRDS: tryptophan, silver nanoparticles, photoreduction, antimicrobial activity, anti-biofilmReCeIV eD: December 20, 2018. ACCePTeD: January 20, 2019.TyPe: Original ResearchFuNDINg: The authors are grateful to the FAPESP (2014/06960-9) and CNPq, Brazilian funding agencies, for their financial support.DeClARATIoN oF C oNFlICTINg INTeReSTS: The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.CoRReSPoNDINg AuTHoR: Lilia Coronato Courrol, Laboratório de Lasers e Óptica Biomédica Aplicada and Departamento de Física, Universidade Federal de São Paulo, Diadema SP 09972-270, Brazil. Email: lccourrol@gmail.com831677TRY0010.1177/1178646919831677International Journal of Tryptophan ResearchCourrol et alresearch-article2019

Tryptophan silver nanoparticles synthesized by photoreduction method: characterization and determination of bactericidal and anti-biofilm activities on resistant and susceptible bacteria

The high rates of antibiotics use in hospitals have resulted in a condition where multidrug-resistant pathogens have become a severe threat to the human health worldwide. Therefore, there is an increasing necessity to identify new antimicrobial agents that can inhibit the multidrug-resistant bacteria and biofilm formation. In this study, antibacterial and anti-biofilm activities of tryptophan silver nanoparticles (TrpAgNP) were investigated. The TrpAgNPs were synthesized by photoreduction method, and the influence of irradiation time and concentration of reagents were analyzed. The nanoparticles were characterized by transmission electron microscopy, Zeta Potential and (UV)-absorption spectra. The antibacterial activity of TrpAgNPs was tested for antibiotic-resistant and susceptible pathogens, Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Citrobacter freundii, Klebsiella pneumoniae, Salmonella typhimurium, and Pseudomonas aeruginosa, evaluating the influence of photoreduction parameters in bactericidal effect. The results have shown that TrpAgNPs solutions with lower tryptophan/silver nitrate (AgNO3) ratio and higher AgNO3 concentration have higher bactericidal action against bacteria with inhibition of ~100% in almost all studied bacterial strains. The antimicrobial activity of TrpAgNPs within biofilms generated under static conditions of antibiotic-resistant and susceptible strains of S. aureus, S. epidermidis, E. coli, K. pneumoniae, C. freundii, and P. aeruginosa was also investigated. The results showed that TrpAgNPs have an inhibitory effect against biofilm formation, exceeding 50% in the case of Gram-negative bacteria (E. coli, K. pneumoniae, C. freundii, and P. aeruginosa—54.8% to 98.8%). For Gram-positive species, an inhibition of biofilm formation of 68.7% to 72.2 % was observed for S. aureus and 20.0% to 40.2% for S. epidermidis.

Novel Hybrid of Typical Enteropathogenic Escherichia coli and Shiga-Toxin-Producing E. coli (tEPEC/STEC) Emerging From Pet Birds.

Exotic psittacine birds have been implicated as reservoir of diarrheagenic Escherichia coli (E. coli), including enteropathogenic E. coli (EPEC) and Shiga-toxin producing E. coli (STEC). Here, we present a genotypic and phenotypic characterization of typical EPEC/STEC hybrid strains isolated from exotic psittacine birds. The strains were positive for eae, bfpA, and stx2f genes, belong to serotype O137:H6 and ST2678. Two strains were subject to whole genome sequencing, confirming the presence of the virulence factors of both E. coli pathotypes. Phenotypical in vitro tests confirmed their ability to adhere to HeLa cells and cause cytotoxicity to Vero cells. The rabbit ileal loop assays showed the attaching and effacing lesion, in addition to inflammatory process and overproduction of intestinal mucus. This is the first report of hybrid typical EPEC/STEC (O137:H6/ST2678) strains isolated from companion psittacine birds and the results suggest zoonotic risks.