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1.
Food Chem ; 398: 133923, 2023 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-35987005

RESUMO

This study aimed to characterize a novel antimicrobial peptide (AMP) obtained from Moringa oleifera seed protein hydrolysates. Cell membrane chromatography and live bacteria adsorption were combined into a single step to efficiently isolate the active fraction of the AMP. Five peptides were identified by LC-MS/MS, among which the MCNDCGA peptide (termed MOp3) showed the greatest inhibitory effect against Staphylococcus aureus [minimum inhibitory concentration (MIC): 2 mg/mL]. MOp3 was identified as a hydrophobic anionic AMP rich in ß-sheet structures with negligible hemolytic activity at 2.0 × MIC. MOp3 had good tolerance to salt solutions at 5 % and pH range 6.0-8.0, but was sensitive to high temperatures (>100 °C) and acid protease. Microscopic observation further revealed that MOp3 induced irreversible damage onto the cell membrane of S. aureus and interacted with dihydrofolate reductase and DNA gyrase by hydrogen bonding and hydrophobic interaction. These findings highlight the potential application of a new antimicrobial agent against S. aureus in the food industry.


Assuntos
Moringa oleifera , Monofosfato de Adenosina/análise , Adsorção , Peptídeos Antimicrobianos , Cromatografia Líquida , Moringa oleifera/química , Extratos Vegetais/química , Sementes/química , Staphylococcus aureus , Espectrometria de Massas em Tandem
2.
Food Chem ; 400: 134030, 2023 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-36055144

RESUMO

The regular release kinetics of active ingredients is inconsistent with the demand of meat preservation. Herein, the pH-triggered dynamic mechanism of packaging film was developed based on L100 polymer incorporated with Cinnamon essential oil (CEO) by coaxial electrospinning. The acquired results revealed that UV-vis and fluorescence spectra confirmed the pH release behavior. The fabricated film was quickly dissolved and transformed from solid to liquid phase, resulting in a faster release rate of CEO from 68.9 % to 98.2 % with the pH increasing. The morphological structure verified the core-shell structure formation with a specific surface area value of 7.22 m2/g. The live/dead results of bacteria indicated good antibacterial efficacy against E. coli and S. aureus. The pH-sensitive film successfully extends the shelf-life of griskin by 3 days. In conclusion, this work will aid in optimizing durability of active ingredients in packaging.


Assuntos
Cinnamomum zeylanicum , Óleos Voláteis , Antibacterianos/química , Cinnamomum zeylanicum/química , Escherichia coli , Embalagem de Alimentos/métodos , Concentração de Íons de Hidrogênio , Carne , Óleos Voláteis/química , Óleos Voláteis/farmacologia , Polímeros , Staphylococcus aureus
3.
Food Chem ; 399: 133970, 2023 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-35998499

RESUMO

Lateral flow immunoassays (LFIAs) are routine methods for rapid foodborne pollutants screening, with detection limits that are closely associated with the label probes used. The exploitation of high performance and robust probe is highly desirable, and remains a great challenge. Herein, we reported an emerging fluorescent nanobeads i.e. carbon-dots (CD) covalently incorporated mesoporous silicon nanoparticles (CD-MSNs) for LFIAs. CD-MSNs revealed brighter fluorescence, larger particle size and more modification sites in comparison with those of single CD. After bio-functionalisation, CD-MSNs probes were introduced to construct LFIA test strips, and designed for ultrasensitive detection of aflatoxin B1 (AFB1) and Staphylococcus aureus (S. aureus), two representative foodborne pollutants, based on the competitive and sandwich models, respectively. Very competitive quantitative detection limits i.e. 0.05 ng/mL and 102 cfu/mL were correspondingly obtained. Additionally, the test strips were successfully applied to rapidly and accurately screen AFB1 and S. aureus in food samples, highlighting their practicality.


Assuntos
Poluentes Ambientais , Nanopartículas , Aflatoxina B1/análise , Carbono , Corantes Fluorescentes , Imunoensaio/métodos , Limite de Detecção , Silício , Staphylococcus aureus
4.
Braz. j. biol ; 83: e243332, 2023. tab, graf
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1345537

RESUMO

Abstract The present study was aimed to manifest the antibacterial and antifungal activity of methanolic extracts of Salix alba L. against seven Gram-positive and Gram-negative bacterial pathogens e.g. Streptococcus pyogenes, Staphylococcus aureus (1), S. aureus (2), Shigella sonnei, Escherichia coli (1), E. coli (2) and Neisseria gonorrhoeae and three fungal isolates from the air such as Aspergillus terreus, A. ornatus, and Rhizopus stolonifer. Two different serotypes of S. aureus and E. coli were used. The agar well-diffusion method results showed the dose-dependent response of plant extracts against bacterial and fungal strains while some organisms were found resistant e.g. E. coli (1), S. sonnei, A. terreus and R. stolonifer. The highest antibacterial activity was recorded at 17.000±1.732 mm from 100 mg/mL of leaves methanolic extracts against S. pyogenes while the activity of most of the pathogens decreased after 24 h of incubation. The highest antifungal activity was reported at 11.833±1.0 mm against A. ornatus at 50 mg/mL after 48 h of the incubation period. These experimental findings endorse the use of S. alba in ethnopharmacological formulations and suggest the use of methanolic extracts of the said plant to develop drugs to control the proliferation of resistant disease causing pathogenic microbes.


Resumo O presente estudo teve como objetivo manifestar a atividade antibacteriana e antifúngica de extratos metanólicos de Salix alba L. contra sete patógenos bacterianos Gram-positivos e Gram-negativos. Streptococcus pyogenes, Staphylococcus aureus (1), S. aureus (2), Shigella sonnei, Escherichia coli (1), E. coli (2) e Neisseria gonorrhoeae e três isolados de fungos do ar, como Aspergillus terreus, A. ornatus, e Rhizopus stolonifer. Dois sorotipos diferentes de S. aureus e E. coli foram usados. Os resultados do método de difusão em ágar mostraram a resposta dependente da dose de extratos de plantas contra cepas de bactérias e fungos, enquanto alguns organismos foram considerados resistentes, e.g. E. coli (1), S. sonnei, A. terreus e R. stolonifer. A maior atividade antibacteriana foi registrada em 17.000 ± 1.732 de 100 mg/mL de extratos metanólicos de folhas contra S. pyogenes, enquanto a atividade da maioria dos patógenos diminuiu após 24 h de incubação. A maior atividade antifúngica foi relatada em 11,833 ± 1,0 contra A. ornatus a 50 mg/mL após 48 h do período de incubação. Esses achados experimentais endossam o uso de S. alba em formulações etnofarmacológicas e sugerem o uso de extratos metanólicos da referida planta para o desenvolvimento de fármacos que controlem a proliferação de doenças resistentes que causam micróbios patogênicos.


Assuntos
Salix , Antifúngicos/farmacologia , Aspergillus , Rhizopus , Staphylococcus aureus , Extratos Vegetais/farmacologia , Testes de Sensibilidade Microbiana , Metanol , Escherichia coli , Antibacterianos/farmacologia
5.
Braz. j. biol ; 83: e247016, 2023. tab, graf
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1339416

RESUMO

Abstract Pheretima posthuma (Vaillant, 1868), a native earthworm of Pakistan and Southeast Asia, has wide utilization in vermicomposting and bioremediation process. In this study, P. posthuma coelomic fluid (PCF) and body paste (PBP) was evaluated as antibacterial agent against ampicillin (AMP) resistant five Gram positive and four Gram negative clinical isolates. The antibacterial effect of different doses (i.e. 25-100 µg/ml) of PCF and PBP along with AMP and azithromycin (AZM) (negative and positive controls, respectively) were observed through disc diffusion and micro-dilution methods. All nine clinical isolates were noticed as AMP resistant and AZM sensitive. Antibacterial effects of PCF and PBP were dose dependent and zone of inhibitions (ZI) against all clinical isolates were between 23.4 ± 0.92 to 0 ± 00 mm. The sensitivity profile of PCF and PBP against clinical isolates was noticed as 44.44 and 55.56%, respectively. Both PCF and PBP showed bacteriostatic (BTS) action against S. aureus, S. pyogenes, K. pneumonia, N. gonorrhoeae. Moreover, the cumulative BTS potential of PCF and PBP against all isolates was 66.67 and 55.56%, respectively. The MICs of PCF and PBP were ranged from 50-200 µg/ml against selected isolates. The bacterial growth curves indicated that PCF and PBP inhibited the growth of all isolates at their specific MIC concentrations. However, PBP has better antibacterial potential compared to PCF against selected isolates. Therefore, it is concluded that both PCF and PBP of P. posthuma possess antibacterial and BTS potential against ampicillin resistant clinical isolates. This organism might be considered as a second choice of antibacterial agents and can further be utilized in pharmaceutical industries for novel drug manufacturing by prospecting bioactive potential agents.


Resumo Pheretima posthuma (Vaillant, 1868), uma minhoca nativa do Paquistão e sudeste da Ásia, tem ampla utilização em processos de vermicompostagem e biorremediação. Neste estudo, o fluido celômico de P. posthuma (PCF) e a pasta corporal (PBP) foram avaliados como agente antibacteriano contra cinco isolados clínicos Gram-positivos e quatro Gram-negativos resistentes à ampicilina (AMP). O efeito antibacteriano de diferentes doses (ou seja, 25-100 µg / ml) de PCF e PBP juntamente com AMP e azitromicina (AZM) (controles negativo e positivo, respectivamente) foi observado por meio de métodos de difusão em disco e microdiluição. Todos os nove isolados clínicos foram notados como resistentes a AMP e sensíveis a AZM. Os efeitos antibacterianos de PCF e PBP foram dependentes da dose e a zona de inibição (ZI) contra todos os isolados clínicos foi entre 23,4 ± 0,92 a 0 ± 00 mm. O perfil de sensibilidade do PCF e PBP contra isolados clínicos foi observado como 44,44% e 55,56%, respectivamente. Tanto o PCF quanto o PBP mostraram ação bacteriostática (BTS) contra S. aureus, S. pyogenes, K. pneumonia, N. gonorrhoeae. Além disso, o potencial BTS cumulativo de PCF e PBP contra todos os isolados foi de 66,67% e 55,56%, respectivamente. Os MICs de PCF e PBP variaram de 50-200 µg / ml contra isolados selecionados. As curvas de crescimento bacteriano indicaram que o PCF e o PBP inibiram o crescimento de todos os isolados em suas concentrações específicas de MIC. No entanto, PBP tem melhor potencial antibacteriano em comparação com PCF contra isolados selecionados. Portanto, conclui-se que tanto o PCF quanto o PBP de P. posthuma possuem potencial antibacteriano e BTS contra isolados clínicos resistentes à ampicilina. Esse organismo pode ser considerado como uma segunda escolha de agentes antibacterianos e pode ainda ser utilizado nas indústrias farmacêuticas para a fabricação de novos medicamentos por meio da prospecção de agentes com potencial bioativo.


Assuntos
Animais , Oligoquetos , Staphylococcus aureus , Testes de Sensibilidade Microbiana , Ampicilina/farmacologia , Antibacterianos/farmacologia
6.
Braz. j. biol ; 83: e245585, 2023. tab, graf
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1339413

RESUMO

Abstract Many soil microorganisms' i.e., bacteria and fungi produce secondary metabolites called antibiotics. These are used for the treatment of some of the bacterial, fungal and protozoal diseases of humans. There is a need for isolation of a broad spectrum of antibiotics from microorganisms due to the emergence of antibiotic resistance. In the present study two antibiotic producing bacteria Klebsiella pneumoniae and Bacillus cereus were isolated from pharmaceutical and poultry feed industry of Hattar, Haripur Pakistan. Total 10 waste samples were collected from different industries (Marble, Ghee, Soap, Mineral, Steel, Poultry Feed, Pharmaceutical, Qarshi, Cosmetic and Glass). Thirty-three bacterial strains were isolated from industrial wastes of these ten different industries. Fourteen out of thirty-three bacterial strains exhibited antimicrobial activities against at least one of the test microbes considered in this study including Escherchia coli, Staphylococcus aureus and Salmonella typhi. The bacteria were isolated by standard serial dilution spread plate technique. Morphological characterization of the isolates was done by Gram staining. Nine bacterial isolates out of fourteen were initially identified as B. cereus and five as K. pneumoniae through biochemical characterization. The antibacterial activities were tested by well diffusion method. Maximum number of antibiotic producing bacteria were isolated from pharmaceutical and poultry feed industry based on the results of primary screening, the most potential isolates S9, S19, S20, S22 and S23 were selected for secondary screening. The maximum activity against E. coli and S. aureus was recorded by bacterial isolate S19 i.e zones of inhibition of 6.5mm and 9mm while S20 showed 7.5mm and 6mm zones respectively. Molecular identification was carried out on the basis of 16S rRNA sequence analysis. Finally, the isolates were identified as B. cereus accession number LC538271and K. pneumoniae accession number MT078679. Analysis of bacterial extract S20 through GC-MS indicated the presence of 8 compounds of diverse nature and structure. Present study suggests that wastes of pharmaceutical and poultry feed industry may have antibiotic producing bacteria. These bacteria could be utilized for the production of antibiotics. B. cereus and K. pneumoniae isolated from wastes of poultry feed and pharmaceutical industries have the potential to produce antibiotics and could be used to control the microbial growth.


Resumo Muitos microrganismos do solo, ou seja, bactérias e fungos produzem metabólitos secundários chamados antibióticos. Eles são usados ​​para tratamento de algumas doenças bacterianas, fúngicas e protozoárias em humanos. Há necessidade de isolamento de um amplo espectro de antibióticos de microrganismos devido ao surgimento de resistência aos antibióticos. No presente estudo, duas bactérias produtoras de antibióticos, Klebsiella pneumoniae e Bacillus cereus, foram isoladas da indústria farmacêutica e de ração avícola de Hattar, Haripur, Paquistão. Um total de 10 amostras de resíduos foi coletado de diferentes indústrias (mármore, ghee, sabão, mineral, aço, ração para aves, farmacêutica, Qarshi, cosmética e vidro). Trinta e três cepas bacterianas foram isoladas de resíduos industriais dessas dez diferentes indústrias. Quatorze das 33 cepas bacterianas exibiram atividades antimicrobianas contra pelo menos um dos micróbios de teste considerados neste estudo, incluindo Escherchia coli, Staphylococcus aureus e Salmonella typhi. As bactérias foram isoladas pela técnica de placa de diluição em série padrão. A caracterização morfológica dos isolados foi feita por coloração de gram. Nove isolados bacterianos de 14 foram inicialmente identificados como B. cereus e cinco como K. pneumoniae por meio de caracterização bioquímica. As atividades antibacterianas foram testadas pelo método de difusão em poço. O número máximo de bactérias produtoras de antibióticos foi isolado da indústria farmacêutica e de ração avícola com base nos resultados da triagem primária, os isolados mais potenciais S9, S19, S20, S22 e S23 foram selecionados para a triagem secundária. A atividade máxima contra E. coli e S. aureus foi registrada pelo isolado bacteriano S19, ou seja, zonas de inibição de 6,5 mm e 9 mm, enquanto S20 mostrou zonas de 7,5 mm e 6 mm, respectivamente. A identificação molecular foi realizada com base na análise da sequência 16S rRNA. Finalmente, os isolados foram identificados como B. cereus número de acesso LC538271 e K. pneumoniae número de acesso MT078679. A análise do extrato bacteriano S20 por meio de GC-MS indicou a presença de oito compostos de natureza e estrutura diversas. O presente estudo sugere que resíduos da indústria farmacêutica e de ração para aves podem conter bactérias produtoras de antibióticos. Essas bactérias podem ser utilizadas para a produção de antibióticos B. cereus e K. pneumoniae isolados de resíduos de rações de aves e indústrias farmacêuticas têm potencial para produzir antibióticos e podem ser usados ​​para controlar o crescimento microbiano.


Assuntos
Humanos , Staphylococcus aureus , Resíduos Industriais , RNA Ribossômico 16S , Extratos Vegetais , Testes de Sensibilidade Microbiana , Escherichia coli , Cromatografia Gasosa-Espectrometria de Massas , Antibacterianos/farmacologia
7.
Braz. j. biol ; 83: e244435, 2023. tab, graf
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1285619

RESUMO

Abstract Increasing trend in antimicrobial resistance and failure of chemically synthesized antibiotics lead to discover alternative methods for the treatment of bacterial infections. Various medicinal plants are in use traditionally and their active compounds can be further applied for treatment of bacterial diseases. This study was designed to determine the antibacterial activity of Punica granatum (P. granatum L.) (pomegranate) peel extract against Enterobacteriaceae [Escherichia coli (E. coli), Salmonella Typhimurium (S. Typhimurium) and Shigella Dysenteriae (S. Dysenteriae)] and gram-positive bacterium [Staphylococcus aureus (Staph aureus)]. Methanolic extract of P. granatum L. peel was prepared by Soxhlet apparatus method. Total flavonoid and phenolic contents from the extract were determined by High Performance Liquid Chromatography (HPLC). The antibacterial activity of P. granatum L. peel extract was evaluated through agar well diffusion method. HPLC showed the range of phenolics (gallic acid, caffeic acid, benzoic acid, cinnamic acid) and flavonoid compounds. The chemical structures of flavonoid and phenolics found in the methanolic extract of P. granatum L. peel have been reported for the first time. The methanolic peel extract (50 ul) of yellow P. granatum L. showed 26, 10, 10 and 9mm zones of inhibition (ZOI) against S. aureus, S. Typhimurium, S. Dysenteriae and E. coli, respectively. The methanolic extract of red P. granatum L. (100 ul) showed 27, 8, 12 and 15 mm ZOI against Staph. aureus, S. Typhimurium, S. Dysenteriae and E. coli, respectively. Highest ZOI was observed against Staph. aureus. Many of the bacteria studied in the present work may cause serious gastrointestinal infections, which can lead to hemorrhagic diarrhea in children. These infections can be life-threatening to young children and the elderly. There is an incentive to find alternative control measures, such as plant and herbal extracts, especially in lesser-developed countries where traditional antibiotics may not be readily available.


Resumo A tendência crescente na resistência antimicrobiana e na falha dos antibióticos sintetizados quimicamente leva à descoberta de métodos alternativos para o tratamento de infecções bacterianas. Várias plantas medicinais estão em uso tradicionalmente e seus compostos ativos podem ser posteriormente aplicados para o tratamento de doenças bacterianas. Este estudo foi desenhado para determinar a atividade antibacteriana do extrato de casca de Punica granatum (P. granatum L.) (romã) contra Enterobacteriaceae [Escherichia coli (E. coli), Salmonella Typhimurium (S. Typhimurium) e Shigella Dysenteriae (S. Dysenteriae) ] e bactéria gram-positiva [Staphylococcus aureus (Staph aureus)]. O extrato metanólico da casca de P. granatum L. foi preparado pelo método do aparelho de Soxhlet. O conteúdo total de flavonoides e fenólicos do extrato foi determinado por cromatografia líquida de alta eficiência (HPLC). A atividade antibacteriana do extrato da casca de P. granatum L. foi avaliada através do método de difusão em ágar. HPLC mostrou a gama de compostos fenólicos (ácido gálico, ácido cafeico, ácido benzoico, ácido cinâmico) e flavonoides. As estruturas químicas de flavonoides e fenólicos encontradas no extrato metanólico da casca de P. granatum L. foram relatadas pela primeira vez. O extrato metanólico da casca (50 ul) de P. granatum L. amarelo apresentou zonas de inibição (ZOI) de 26, 10, 10 e 9mm contra S. aureus, S. Typhimurium, S. Dysenteriae e E. coli, respectivamente. O extrato metanólico de P. granatum L. vermelho (100 ul) apresentou 27, 8, 12 e 15 mm IOI contra Staph. aureus, S. Typhimurium, S. Dysenteriae e E. coli, respectivamente. O ZOI mais alto foi observado contra Staph. aureus. Muitas das bactérias estudadas no presente trabalho podem causar infecções gastrointestinais graves, que podem levar à diarreia hemorrágica em crianças. Essas infecções podem ser fatais para crianças pequenas e idosos. Há um incentivo para encontrar medidas de controle alternativas, como extratos de plantas e ervas, especialmente em países menos desenvolvidos, onde os antibióticos tradicionais podem não estar prontamente disponíveis.


Assuntos
Humanos , Pré-Escolar , Criança , Idoso , Romã (Fruta) , Staphylococcus aureus , Extratos Vegetais/farmacologia , Cromatografia Líquida de Alta Pressão , Escherichia coli , Anti-Infecciosos
8.
Braz. j. biol ; 83: e245807, 2023. tab, graf
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1285613

RESUMO

Abstract Linum usitatissimum L is a widely used traditionally for multiple ailments. The present research was carried out to explore the antimicrobial, and anti-biofilm activity of crude extract of Linum usitatissimum L (Lu. Cr). Phytochemical and proximate analyses were performed. The bandages of diabetic foot patients were collected from the various hospitals. The bandages were cultured to isolate the bacterial strains present on it. The disc diffusion method was used to identify the antimicrobial potential whereas the minimum inhibitory concentration of the Lu.Cr were also determined. Proximate analysis confirms moisture content 8.33%, ash content 4.33%, crude protein 21.20%, crude fat 49.2% and crude fiber 5.63%. It was revealed that Gram-positive bacteria are most prevalent among all study groups. Lu.Cr possess significant bactericidal potential against S. aureus among all other microbes. Owing to this potential, linseed coated bandages can be used alternatively for the treatment of diabetic foot.


Resumo Linum usitatissimum L é amplamente utilizado tradicionalmente para doenças múltiplas. O presente trabalho foi realizado para explorar a atividade antimicrobiana e antibiofilme do extrato bruto de Linum usitatissimum L (Lu.Cr). Foram realizadas análises fitoquímicas e aproximadas. As ataduras de pacientes diabéticos com pé foram recolhidas nos vários hospitais. As bandagens foram cultivadas para isolar as cepas bacterianas presentes nas mesmas. O método de difusão em disco foi utilizado para identificar o potencial antimicrobiano e a concentração inibitória mínima do Lu.Cr também foi determinada. A análise aproximada confirma o teor de umidade 8,33%, teor de cinzas 4,33%, proteína bruta 21,20%, gordura bruta 49,2% e fibra bruta 5,63%. Foi revelado que as bactérias Gram-positivas são mais prevalentes entre todos os grupos de estudo. Lu.Cr possui potencial bactericida significativo contra S. aureus entre todos os outros micróbios. Devido a esse potencial, as ligaduras revestidas com linhaça podem ser utilizadas alternativamente para o tratamento do pé diabético.


Assuntos
Humanos , Pé Diabético/tratamento farmacológico , Linho , Diabetes Mellitus , Staphylococcus aureus , Extratos Vegetais/farmacologia , Testes de Sensibilidade Microbiana , Biofilmes , Metanol
9.
J Adv Res ; 40: 167-178, 2022 09.
Artigo em Inglês | MEDLINE | ID: mdl-36100324

RESUMO

INTRODUCTION: Vancomycin-intermediate Staphylococcus aureus (VISA) is typically associated with a decline in virulence. We previously reported a WalK(S221P) mutation that plays an important role in mediating vancomycin resistance in VISA XN108. Whether this mutation is implicated in bacterial virulence remains unknown. OBJECTIVES: This study aimed to investigate the effect of WalK(S221P) mutation on the virulence of VISA and the underlying mechanism of this effect. METHODS: The influence of WalK(S221P) mutation on VISA virulence and its underlying mechanism were explored using animal models, RNA-seq analysis, RT-qPCR, hemolytic assay, slide coagulase test, Western blot, ß-galactosidase assay, and electrophoresis mobility shift assay (EMSA). RESULTS: Compared with XN108, WalK(S221P)-reverted strain XN108-R exacerbated cutaneous infections with increased lesion size and extensive inflammatory infiltration in mouse models. The bacterial loads of S. aureus XN108-R in murine kidney increased compared with those of XN108. RNA-seq analysis showed upregulation of a set of virulence genes in XN108-R, which exhibited greater hemolytic and stronger coagulase activities compared with XN108. Introduction of WalK(S221P) to methicillin-resistant S. aureus USA300 and methicillin-susceptible strain Newman increased the vancomycin resistance of the mutants, which exhibited reduced hemolytic activities and decreased expression levels of many virulence factors compared with their progenitors. WalK(S221P) mutation weakened agr promoter-controlled ß-galactosidase activity. EMSA results showed that WalK-phosphorylated WalR could directly bind to the agr promoter region, whereas WalK(S221P)-activated WalR reduced binding to the target promoter. Inactivation of agr in S. aureus did not affect their vancomycin susceptibility but mitigated the virulence alterations caused by WalK(S221P) mutation. CONCLUSION: The results of our study indicate that WalK(S221P) mutation can enhance vancomycin resistance in S. aureus of diverse genetic backgrounds. WalK(S221P)- bearing S. aureus strains exhibit reduced virulence. WalK(S221P) mutation may directly impair the activation of the agr system by WalR, thereby decreasing the expression of virulence factors in VISA.


Assuntos
Staphylococcus aureus Resistente à Meticilina , Resistência a Vancomicina , Animais , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/farmacologia , Coagulase/genética , Coagulase/farmacologia , Staphylococcus aureus Resistente à Meticilina/genética , Staphylococcus aureus Resistente à Meticilina/metabolismo , Camundongos , Testes de Sensibilidade Microbiana , Mutação , Staphylococcus aureus/genética , Resistência a Vancomicina/genética , Staphylococcus aureus Resistente à Vancomicina , Virulência/genética , Fatores de Virulência/genética , Fatores de Virulência/farmacologia , beta-Galactosidase/genética , beta-Galactosidase/farmacologia
10.
Antimicrob Resist Infect Control ; 11(1): 114, 2022 09 14.
Artigo em Inglês | MEDLINE | ID: mdl-36104710

RESUMO

BACKGROUND: Staphylococcus aureus is one of the leading causes of bloodstream infections (BSI) worldwide. In Brazil, the hospital-acquired methicillin-resistant S. aureus USA100/SCCmecII lineage replaced the previously well-established clones. However, the emergence of community-associated (CA) MRSA lineages among hospitalized patients is an increasing issue. METHODS: Consecutive S. aureus isolates recovered from BSI episodes of patients admitted between January 2016 and December 2018 in a Brazilian teaching hospital were tested for antimicrobial resistance, their genotypic features were characterized, and the clinical characteristics of the patients were evaluated. RESULTS: A total of 123 S. aureus isolates were recovered from 113 patients. All isolates were susceptible to linezolid, teicoplanin and vancomycin and 13.8% were not susceptible to daptomycin. Vancomycin MIC50 and MIC90 of 2 mg/L were found for both MRSA and MSSA isolates. The MRSA isolation rate was 30.1% (37/123), and 51.4% of them carried the SCCmec type II, followed by SCCmecIV (40.5%). Among the 37 MRSA isolates, the main lineages found were USA100/SCCmecII/ST5 and ST105 (53.7%) and USA800/ST5/SCCmecIV (18.9%). Surprisingly, six (16%) CA-MRSA isolates, belonging to USA300/ST8/SCCmecIVa that carried PVL genes and the ACME cassette type I, were detected. These six patients with USA300 BSI had severe comorbidities, including cancer, and most had a Charlson score ≥ 5; furthermore, they were in wards attended by the same health professionals. MRSA isolates were associated with hospital acquired infections (p = 0.02) in more elderly patients (p = 0.03) and those diagnosed with hematologic cancer (p = 0.04). Among patients diagnosed with MRSA BSI, 19 (54.3%) died. CONCLUSIONS: The pandemic MRSA USA300 was detected for the first time in the Brazilian teaching hospital under study, and its cross-transmission most probably occurred between patients with BSI. This lineage may already be circulating among other Brazilian hospitals, which highlights the importance of carrying out surveillance programs to fight multidrug resistant and hypervirulent isolates.


Assuntos
Staphylococcus aureus Resistente à Meticilina , Sepse , Idoso , Brasil/epidemiologia , Células Clonais , Hospitais , Humanos , Pandemias , Staphylococcus aureus , Vancomicina
11.
J Nanobiotechnology ; 20(1): 416, 2022 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-36109760

RESUMO

BACKGROUND: Drug resistance of pathogens and immunosuppression are the main causes of clinical stagnation of osteomyelitis. The ideal treatment strategy for osteomyelitis is to achieve both efficient antibacterial and bone healing through spatiotemporal modulation of immune microenvironment. METHODS: In this study, a bilayer hydrogel based on genetically engineered polypeptide AC10A and AC10ARGD was prepared by self-assembly. Ag2S QDs@DSPE-mPEG2000-Ce6/Aptamer (AD-Ce6/Apt) was loaded in the top layer AC10A hydrogel (AA) for antibacterial, and bone marrow-derived mesenchymal stem cells (BMSCs) were loaded in the lower layer AC10ARGD hydrogel (MAR) for bone healing. The AD-Ce6/Apt can be released from the AA hydrogel to target S. aureus before bacterial biofilm formation and achieved significant bactericidal effect under irradiation with a 660 nm laser. Moreover, AD-Ce6/Apt can induce M1 type polarization of macrophages to activate the immune system and eliminate residual bacteria. Subsequently, BMSCs released from the MAR hydrogel can differentiate into osteoblasts and promote the formation of an anti-inflammatory microenvironment by regulating the M2 type polarization of macrophages. The bilayer AA-MAR hydrogel possessed good biocompatibility. RESULTS: The in vitro and in vivo results showed that the AA-MAR hydrogel not only realized efficient photodynamic therapy of S. aureus infection, but also promoted the transformation of immune microenvironment to fulfill the different needs of each stage, which ultimately improved bone regeneration and mechanical properties post-surgery. CONCLUSION: This work presents an approach for spatiotemporal modulation of immune microenvironment in the treatment of osteomyelitis.


Assuntos
Hidrogéis , Osteomielite , Antibacterianos/química , Antibacterianos/farmacologia , Dimaprit/análogos & derivados , Humanos , Hidrogéis/química , Osteomielite/tratamento farmacológico , Peptídeos/farmacologia , Staphylococcus aureus
12.
BMC Microbiol ; 22(1): 219, 2022 09 17.
Artigo em Inglês | MEDLINE | ID: mdl-36115948

RESUMO

BACKGROUND: The prevalence of Staphylococcus aureus isolates carrying the Panton-Valentine leukocidin (PVL) gene is higher in Africa (≈50%) compared to Europe (< 5%). The study aimed to measure anti-PVL-antibodies in Africans and Germans in a multi-center study and to test whether detected antibodies can neutralize the cytotoxic effect of PVL on polymorphonuclear leukocytes (PMNs). METHODS: Sera from asymptomatic Africans (n = 22, Nigeria, Gabon) and Caucasians (n = 22, Germany) were used to quantify antibody titers against PVL and α-hemolysin (in arbitrary units [AU]) by ELISA. PMNs from one African and German donor were exposed to 5 nM recombinant PVL to measure the neutralizing effect of serial dilutions of pooled sera from African and Caucasian participants, or donor sera at 0.625 and 2.5% (v/v). RESULTS: Anti-PVL-antibodies were significantly higher in Africans than in Germans (1.9 vs. 0.7 AU, p < 0.0001). The pooled sera from the study participants neutralized the cytotoxic effect of PVL on African and German PMNs in a dose dependent manner. Also, neutralization of PVL on PMNs from the African and German donors had a stronger effect with African sera (half-maximal inhibitory concentration (IC50) = 0.27 and 0.47%, respectively) compared to Caucasian sera (IC50 = 3.51 and 3.59% respectively). CONCLUSION: Africans have higher levels of neutralizing anti-PVL-antibodies. It remains unclear if or at what level these antibodies protect against PVL-related diseases.


Assuntos
Infecções Estafilocócicas , Staphylococcus aureus , Toxinas Bacterianas , Exotoxinas , Proteínas Hemolisinas , Humanos , Leucocidinas , Nigéria , Infecções Estafilocócicas/epidemiologia
13.
Drug Deliv ; 29(1): 3035-3050, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36120935

RESUMO

Quercetin (QT) is a flavonoid that exhibits anti-oxidant and chemo-preventive activity. This research work aimed to develop surface-modified bilosomes (BS) of QT. The BS was prepared by the solvent evaporation method and optimized by the Box-Behnken design. The optimized QT-BS (QT-BS3opt) displayed vesicle size (143.51 nm), PDI (0.256), zeta potential (-15.4 mV), and entrapment efficiency (89.52%). Further, the optimized QT-BS formulation was coated with chitosan (CS). The XRD diffractogram of CS-QT-BS3opt1 did not exhibit extensive peaks of QT, revealing that QT is properly encapsulated in the polymer matrix. The QT-BS3opt and CS-QT-BS3opt1 exhibited sustained-release (86.62 ± 3.23% and 69.32 ± 2.57%, respectively) up to 24 h with the Korsmeyer-Peppas kinetic model (R2 =0.9089). CS-QT-BS3opt1 exhibited significantly (P < .05) high flux, i.e. 4.20-fold more than pure QT dispersion and 1.27-fold higher than QT-BS3opt. CS-QT-BS3opt1 showed significantly greater bio-adhesion (76.43 ± 2.42%) than QT-BS3opt (20.82 ± 1.45%). The antioxidant activity showed that QT from CS-QT-BS3opt1 has more remarkable (P < .05) antioxidant activity at each concentration than pure QT. The CS-QT-BS3opt1 exhibited 1.61-fold higher cytotoxicity against MFC7 and 1.44-fold higher cytotoxicity against MDA-MB-231 than pure QT. The CS-QT-BS3opt1 displayed a significantly greater antimicrobial potential against E. coli than against S. aureus. From all these findings, it could be concluded that surface-modified QT-BS might be an effective approach for increasing the efficacy of QT in the treatment of certain ailments.


Assuntos
Anti-Infecciosos , Quitosana , Antibacterianos/farmacologia , Antioxidantes/farmacologia , Preparações de Ação Retardada , Escherichia coli , Polímeros , Quercetina/farmacologia , Solventes , Staphylococcus aureus
14.
Commun Biol ; 5(1): 910, 2022 Sep 05.
Artigo em Inglês | MEDLINE | ID: mdl-36065015

RESUMO

Phenol-soluble modulin α (PSMα) is identified as potent virulence factors in Staphylococcus aureus (S. aureus) infections. Very little is known about the role of PSMß which belongs to the same toxin family. Here we compared the role of PSMs in S. aureus-induced septic arthritis in a murine model using three isogenic S. aureus strains differing in the expression of PSMs (Newman, Δpsmα, and Δpsmß). The effects of PSMs on neutrophil NADPH-oxidase activity were determined in vitro. We show that the PSMα activates neutrophils via the formyl peptide receptor (FPR) 2 and reduces their NADPH-oxidase activity in response to the phorbol ester PMA. Despite being a poor neutrophil activator, PSMß has the ability to reduce the neutrophil activating effect of PSMα and to partly reverse the effect of PSMα on the neutrophil response to PMA. Mice infected with S. aureus lacking PSMα had better weight development and lower bacterial burden in the kidneys compared to mice infected with the parental strain, whereas mice infected with bacteria lacking PSMß strain developed more severe septic arthritis accompanied with higher IL-6 and KC. We conclude that PSMα and PSMß play distinct roles in septic arthritis: PSMα aggravates systemic infection, whereas PSMß protects arthritis development.


Assuntos
Artrite Infecciosa , Infecções Estafilocócicas , Animais , Toxinas Bacterianas , Camundongos , NADP/metabolismo , Oxirredutases/metabolismo , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/fisiologia
15.
PLoS Pathog ; 18(9): e1010766, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-36067266

RESUMO

Wound infections are often polymicrobial in nature, biofilm associated and therefore tolerant to antibiotic therapy, and associated with delayed healing. Escherichia coli and Staphylococcus aureus are among the most frequently cultured pathogens from wound infections. However, little is known about the frequency or consequence of E. coli and S. aureus polymicrobial interactions during wound infections. Here we show that E. coli kills Staphylococci, including S. aureus, both in vitro and in a mouse excisional wound model via the genotoxin, colibactin. Colibactin biosynthesis is encoded by the pks locus, which we identified in nearly 30% of human E. coli wound infection isolates. While it is not clear how colibactin is released from E. coli or how it penetrates target cells, we found that the colibactin intermediate N-myristoyl-D-Asn (NMDA) disrupts the S. aureus membrane. We also show that the BarA-UvrY two component system (TCS) senses the environment created during E. coli and S. aureus mixed species interaction, leading to upregulation of pks island genes. Further, we show that BarA-UvrY acts via the carbon storage global regulatory (Csr) system to control pks expression. Together, our data demonstrate the role of colibactin in interspecies competition and show that it is regulated by BarA-UvrY TCS during interspecies competition.


Assuntos
Infecções por Escherichia coli , Proteínas de Escherichia coli , Policetídeos , Infecção dos Ferimentos , Animais , Antibacterianos/metabolismo , Carbono/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Proteínas de Escherichia coli/metabolismo , Humanos , Proteínas de Membrana/metabolismo , Camundongos , Mutagênicos/metabolismo , N-Metilaspartato/metabolismo , Peptídeos , Fosfotransferases/genética , Policetídeos/metabolismo , Staphylococcus/metabolismo , Staphylococcus aureus/genética , Staphylococcus aureus/metabolismo , Fatores de Transcrição/metabolismo
16.
Arch Microbiol ; 204(9): 584, 2022 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-36048277

RESUMO

A tannase-positive Bacillus gottheilii M2S2 and Bacillus cereus M1GT were co-cultivated for the production of gallic acid using tannic acid as the sole carbon source through submerged fermentation. Taguchi orthogonal array of design of experimental methodology was used to estimate the influence and significance of tannic acid concentration, glucose concentration, agitation speed, and inoculum size on the gallic acid production in a shake flask. Among all the factors, agitation speed contributed the highest for gallic acid production (28.28%), followed by glucose concentration (21.59%), inoculum size (19.6%), tannic acid concentration (19.54%), and pH (11.09%). Validation experiments were executed at the found optimized conditions which resulted in a 6.36-fold increase in gallic acid yield compared to unoptimized conditions. Further, the kinetics of growth, tannic acid degradation, and gallic acid yield were evaluated at the optimized conditions. The kinetic parameters Y x/s, Y p/s, and Y p/x were determined as 0.292 mg of cells/mg of tannic acid, 22.2 µg of gallic acid/mg of tannic acid, and 70.76 µg of gallic acid/mg of cells with a growth rate of 0.273 h -1 after 24 h of fermentation. Finally, the antimicrobial activity of the product gallic acid was investigated against food-borne pathogenic E. coli, S. aureus, and Serriatia marcescens and showed a zone of inhibition of 2 cm, 1.6 cm, and 1.3 cm, respectively, using the agar disc diffusion technique. Thus, the cost-effective bioproduct gallic acid proved to be potentially effective to control food poisoning diseases and preserve foodstuff.


Assuntos
Anti-Infecciosos , Ácido Gálico , Anti-Infecciosos/farmacologia , Técnicas de Cocultura , Escherichia coli/metabolismo , Fermentação , Ácido Gálico/metabolismo , Ácido Gálico/farmacologia , Glucose , Cinética , Staphylococcus aureus/metabolismo , Taninos/metabolismo , Taninos/farmacologia
17.
PLoS One ; 17(9): e0273790, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36048838

RESUMO

In Ethiopia, the bacteriological quality of ready-to-eat raw meat is of a great public health concern as it can serve as a source of meat-borne pathogens and worsen the transmission of antimicrobial resistant bacteria, and hence this cross-sectional study, done on 257 meat samples (ie., 169 beef, 50 mutton and 38 chevon) from randomly selected hotels and restaurants (n = 52). Approximately 25 gm of meat samples were taken bi-weekly and subjected to quantitative and qualitative analyses; antimicrobial susceptibility tests were done as per the Kirby-Bauer disk diffusion method. It was found that 13.2 (n = 34), 17.5 (n = 45) and 21.8% (n = 56) samples exceeded the permissible limit for total viable and coliform and S. aureus counts, respectively. At the same time, 24.9% (n = 64) surpassed the bacteriological limit permissible for consumption. Overall, 36.6% (n = 94) of samples were extrapolated as unsatisfactory for consumption due to high bacterial load and or the presence of pathogens. Five different bacterial spp. such as E. coli 65% (n = 167), S. aureus 59% (n = 152), Salmonella spp. 28.4% (n = 73), Campylobacter spp. 14.4% (n = 37) and Shigella spp. 4.3% (n = 11) were isolated in varied proportions. Alarmingly, 60% (n = 264) of the isolates were multi-drug resistant and 51% of S. aureus were found to be MRSA.


Assuntos
Escherichia coli , Restaurantes , Animais , Antibacterianos/farmacologia , Bovinos , Estudos Transversais , Etiópia , Carne/microbiologia , Testes de Sensibilidade Microbiana , Staphylococcus aureus
18.
Sci Rep ; 12(1): 15359, 2022 Sep 13.
Artigo em Inglês | MEDLINE | ID: mdl-36100625

RESUMO

In this work, Ag-ZnO nanocomposites were prepared by a green synthesis route using aqueous leaf extract of Tetradenia riperia and investigated for antibacterial activity against Escherichia coli and Staphylococcus aureus. To optimize the synthesis of the Ag-ZnO, the effects of precursor concentrations, pH, and temperatures were studied. The Ag-ZnO nanocomposites were characterized by XRD, ATR-FTIR, FESEM, and TEM. Results show that the concentration of 8% Ag, the temperature of 80 °C, and a pH of 7-8 were optimal for the synthesis of Ag-ZnO nanocomposites. The XRD analysis showed the decrease in particle size of Ag-ZnO from 23.6 to 14.8 nm with an increase in Ag concentrations, which was further supported by FESEM analysis. TEM image of 8% Ag provides more information on the coexistence of Ag on ZnO where an average particle size of 14.8 nm was determined. The ATR-FTIR analysis confirmed the presence of phenolic compounds, which work as reducing and stabilizing agents. The antimicrobial activity results show that Ag-ZnO nanocomposite demonstrated a higher antimicrobial potency on E. coli than on S. aureus. Therefore, Tetradenia riperia leaf extract is a viable route for the synthesis of Ag-ZnO nanocomposites to be used for various applications, including water disinfection.


Assuntos
Anti-Infecciosos , Lamiaceae , Óxido de Zinco , Antibacterianos/química , Anti-Infecciosos/química , Anti-Infecciosos/farmacologia , Desinfecção , Escherichia coli , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Prata/química , Prata/farmacologia , Staphylococcus aureus , Água/farmacologia , Óxido de Zinco/química , Óxido de Zinco/farmacologia
19.
Pak J Biol Sci ; 25(7): 602-607, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-36098166

RESUMO

<b>Background and Objective:</b> <i>Kalanchoe tomentosa</i> is identified and their different characteristics regarding the antibacterial and antioxidant properties have a vast effect. Fresh <i>K. tomentosa</i> leaves obtained from Bandung, Indonesia was extracted using n-hexane followed by serial dichloromethane maceration. <b>Materials and Methods:</b> N-hexane and ethyl acetate were used to separate the dichloromethane extract using vacuum liquid chromatography and the isolated compounds were recrystallized with n-hexane. <b>Results:</b> About 37 mg of dichloromethane extract was obtained from the extraction process. Recrystallized compound isolates were identified as stigmast-5-en-3-ol or ß-sitosterol. Both dichloromethane extract and ß-sitosterol isolated compounds showed strong bacteriostatic activity against <i>S. aureus</i> with MIC = 15.63 and 7.81 µg mL<sup></sup><sup>1</sup> and<i> K. pneumonia</i> with MIC = 7.81 and 31.25 µg mL<sup></sup><sup>1</sup>, respectively. However, only dichloromethane extract exhibited a bactericidal effect (7.81 µg mL<sup></sup><sup>1</sup>). <b>Conclusion:</b> The pure ß-sitosterol compound was isolated from<i> K. tomentosa</i> dichloromethane extract. Both the dichloromethane extract and the isolated ß-sitosterol compound had antibacterial effects against <i>S. aureus</i> and <i>K. pneumonia.</i>.


Assuntos
Kalanchoe , Pneumonia , Antibacterianos/química , Antibacterianos/farmacologia , Klebsiella , Cloreto de Metileno , Extratos Vegetais/química , Folhas de Planta/química , Sitosteroides , Staphylococcus aureus
20.
World J Microbiol Biotechnol ; 38(12): 221, 2022 Sep 13.
Artigo em Inglês | MEDLINE | ID: mdl-36097302

RESUMO

Mastitis is a significant disease in dairy ruminants, causing economic losses to the livestock industry and severe risks to public health. Antibiotic therapy is one of the most crucial practices to treat mastitis, although the susceptibility of caprine mastitis pathogens to current antibiotics has not been tested under standard or modified incubation conditions. This work evaluated the in vitro activity of tildipirosin, gamithromycin, oxytetracycline, and danofloxacin against caprine mastitis pathogens incubated following standard conditions of Clinical and Laboratory Standards Institute (CLSI) and deviation method by 25% supplementation with goat serum. Mycoplasma agalactiae, Escherichia coli, Staphylococcus aureus, Streptococcus spp., and coagulase-negative Staphylococci (CNS) were isolated from dairy goats with mastitis in Spain. Minimum inhibitory concentrations (MICs) were determined using the broth microdilution technique. The lowest MIC90 under standard conditions was obtained with danofloxacin for mastitis-causing pathogens. An exception was M. agalactiae, where danofloxacin and oxytetracycline obtained low values. However, after adding serum, gamithromycin showed the lowest MIC50 for S. aureus, Streptococcus spp., and CNS. The lowest MIC50 was obtained with all the antibiotics tested (< 0.125 µg/ml) against M. agalactiae. Supplementing with serum resulted in a significant variation in tildipirosin and gamithromycin MIC values for CNS, S. aureus, M. agalagtiae, and E. coli. In brief, the MIC for antibiotics used against mastitis should be determined under conditions closely resembling intramammary infections to obtain representative susceptibility patterns against mastitis pathogens. Caprine mastitis pathogens were broadly susceptible to danofloxacin under standard conditions. The potency of macrolides against caprine mastitis pathogens increases when serum is present in culture media.


Assuntos
Mastite Bovina , Oxitetraciclina , Animais , Antibacterianos/farmacologia , Bovinos , Escherichia coli , Feminino , Fluoroquinolonas , Cabras , Humanos , Macrolídeos , Mastite Bovina/tratamento farmacológico , Oxitetraciclina/farmacologia , Oxitetraciclina/uso terapêutico , Staphylococcus , Staphylococcus aureus , Streptococcus , Tilosina/análogos & derivados , Tilosina/farmacologia , Tilosina/uso terapêutico
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