Phytochemical characterization, antioxidant potential and antibacterial activity of the Croton argyrophylloides Muell. Arg. (Euphorbiaceae) / Caracterização fitoquímica, potencial antioxidante e atividade antibacteriana do Croton argyrophylloides Muell. Arg. (Euphorbiaceae)

Abstract Croton argyrophylloides Muell. Arg., from the Euphorbiaceae family, popularly known as marmeleiro prateado or sacatinga, is a plant from the Caatinga biome commonly found in Brazil's northeastern region. The present study aimed to evaluate the antioxidant activity of the species. The phytochemical study was performed through qualitative analysis of chemical constituents and quantitative determination of the total phenol content through the Folin-Ciocalteu test. The qualitative and quantitative antioxidant tests were performed using the DPPH method (2.2 diphenyl-1-picryl hydrazil) and ferric reducing antioxidant power (FRAP). The minimum inhibitory concentration (MIC) was determined by microdilution in 96-well plates. The ethanolic extract of the leaves of C. argyrophylloides manifested antioxidant action in the quantitative DPPH test with a significant bioactivity of 84.70 AAO% in 500 µg/mL, with an EC50 of 236.79. The content of total phenolic compounds was 946.06 mg of gallic acid equivalents/g of sample, and total flavonoids was 58.11 mg of quercetin equivalents/g of sample, the result obtained for FRAP was 15294.44 µM Trolox/g of sample and ABTS was 718 μM Trolox of sample. The prospecting of the chemical constituents of the leaves of C. argyrophylloides revealed the presence of the main compounds that manifests the antioxidant activity and it was proven by the DPPH method that there is antioxidant activity in the analyzed sample, in addition to demonstrating a significant content of phenolic compounds and total flavonoid content in the species, which corroborates the antioxidant activity of the plant sample. The leaf extracts presented growth inhibition halos of 10 and 12 mm upon Staphylococcus aureus ATCC 25923.

Resumo Croton argyrophylloides Muell. Arg., pertencente à família Euphorbiaceae, conhecida popularmente como marmeleiro prateado e sacatinga, é um vegetal do bioma caatinga comumente encontrado no Nordeste do Brasil. O presente trabalho teve como objetivo avaliar a atividade antioxidante da espécie. O estudo fitoquímico foi realizado por meio de análise qualitativa dos constituintes químicos e determinação quantitativa do teor de fenóis totais pelo teste de Folin-Ciocalteu. Os testes antioxidantes qualitativos e quantitativos foram realizados pelo método do DPPH (2,2 difenil-1- picril-hidrazila) e redução do ferro (FRAP). A concentração inibitória mínima (CIM) foi determinada por microdiluição em placas de 96 poços. O extrato etanólico das folhas de C. argyrophylloides apresentou ação antioxidante no teste DPPH quantitativo com uma significativa bioatividade de 84.70 AAO% em 500 µg/mL, apresentando um CE50 de 236.79. O teor de compostos fenólicos totais, foi de 946,06 mg equivalentes de ácido gálico/g de amostra, e de flavonoides totais de 58,11 mg equivalentes de quercetina/g da amostra, o valor encontrado para FRAP foi de 15294,44 µM Trolox/g da amostra e de ABTS foi 718 μM Trolox da amostra. A prospecção dos constituintes químicos das folhas de C. argyrophylloides revelou a presença dos principais compostos que caracterizam a atividade antioxidante e foi possível comprovar pelo método de DPPH que há atividade antioxidante na amostra analisada, além de demonstrar um resultado significativo de teor de compostos fenólicos e teor de flavonoides totais na espécie e o que corrobora com a atividade antioxidante da amostra vegetal. Os extratos das folhas apresentaram halos de inibição de crescimento de 10 e 12mm frente a Staphylococcus aureus ATCC 25923.

Evaluation of methanolic crude extract of Linum usitatissimum for the removal of biofilm in diabetic foot isolates / Avaliação do extrato em bruto metanólico de Linum usitatissimum para remoção de biofilmes em isolados diabéticos de pé

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.

Evaluation of methanolic crude extract of Linum usitatissimum for the removal of biofilm in diabetic foot isolates / Avaliação do extrato em bruto metanólico de Linum usitatissimum para remoção de biofilmes em isolados diabéticos de pé

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.

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.

Phytochemical characterization, antioxidant potential and antibacterial activity of the Croton argyrophylloides Muell. Arg. (Euphorbiaceae) / Caracterização fitoquímica, potencial antioxidante e atividade antibacteriana do Croton argyrophylloides Muell. Arg. (Euphorbiaceae)

Croton argyrophylloides Muell. Arg., from the Euphorbiaceae family, popularly known as marmeleiro prateado or sacatinga, is a plant from the Caatinga biome commonly found in Brazil's northeastern region. The present study aimed to evaluate the antioxidant activity of the species. The phytochemical study was performed through qualitative analysis of chemical constituents and quantitative determination of the total phenol content through the Folin-Ciocalteu test. The qualitative and quantitative antioxidant tests were performed using the DPPH method (2.2 diphenyl-1-picryl hydrazil) and ferric reducing antioxidant power (FRAP). The minimum inhibitory concentration (MIC) was determined by microdilution in 96-well plates. The ethanolic extract of the leaves of C. argyrophylloides manifested antioxidant action in the quantitative DPPH test with a significant bioactivity of 84.70 AAO% in 500 µg/mL, with an EC50 of 236.79. The content of total phenolic compounds was 946.06 mg of gallic acid equivalents/g of sample, and total flavonoids was 58.11 mg of quercetin equivalents/g of sample, the result obtained for FRAP was 15294.44 µM Trolox/g of sample and ABTS was 718 µM Trolox of sample. The prospecting of the chemical constituents of the leaves of C. argyrophylloides revealed the presence of the main compounds that manifests the antioxidant activity and it was proven by the DPPH method that there is antioxidant activity in the analyzed sample, in addition to demonstrating a significant content of phenolic compounds and total flavonoid content in the species, which corroborates the antioxidant activity of the plant sample. The leaf extracts presented growth inhibition halos of 10 and 12 mm upon Staphylococcus aureus ATCC 25923.

Croton argyrophylloides Muell. Arg., pertencente à família Euphorbiaceae, conhecida popularmente como marmeleiro prateado e sacatinga, é um vegetal do bioma caatinga comumente encontrado no Nordeste do Brasil. O presente trabalho teve como objetivo avaliar a atividade antioxidante da espécie. O estudo fitoquímico foi realizado por meio de análise qualitativa dos constituintes químicos e determinação quantitativa do teor de fenóis totais pelo teste de Folin-Ciocalteu. Os testes antioxidantes qualitativos e quantitativos foram realizados pelo método do DPPH (2,2 difenil-1- picril-hidrazila) e redução do ferro (FRAP). A concentração inibitória mínima (CIM) foi determinada por microdiluição em placas de 96 poços. O extrato etanólico das folhas de C. argyrophylloides apresentou ação antioxidante no teste DPPH quantitativo com uma significativa bioatividade de 84.70 AAO% em 500 µg/mL, apresentando um CE50 de 236.79. O teor de compostos fenólicos totais, foi de 946,06 mg equivalentes de ácido gálico/g de amostra, e de flavonoides totais de 58,11 mg equivalentes de quercetina/g da amostra, o valor encontrado para FRAP foi de 15294,44 µM Trolox/g da amostra e de ABTS foi 718 µM Trolox da amostra. A prospecção dos constituintes químicos das folhas de C. argyrophylloides revelou a presença dos principais compostos que caracterizam a atividade antioxidante e foi possível comprovar pelo método de DPPH que há atividade antioxidante na amostra analisada, além de demonstrar um resultado significativo de teor de compostos fenólicos e teor de flavonoides totais na espécie e o que corrobora com a atividade antioxidante da amostra vegetal. Os extratos das folhas apresentaram halos de inibição de crescimento de 10 e 12mm frente a Staphylococcus aureus ATCC 25923.

Evaluation of methanolic crude extract of Linum usitatissimum for the removal of biofilm in diabetic foot isolates

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.

Phytochemical characterization, antioxidant potential and antibacterial activity of the Croton argyrophylloides Muell. Arg. (Euphorbiaceae)

Abstract Croton argyrophylloides Muell. Arg., from the Euphorbiaceae family, popularly known as marmeleiro prateado or sacatinga, is a plant from the Caatinga biome commonly found in Brazils northeastern region. The present study aimed to evaluate the antioxidant activity of the species. The phytochemical study was performed through qualitative analysis of chemical constituents and quantitative determination of the total phenol content through the Folin-Ciocalteu test. The qualitative and quantitative antioxidant tests were performed using the DPPH method (2.2 diphenyl-1-picryl hydrazil) and ferric reducing antioxidant power (FRAP). The minimum inhibitory concentration (MIC) was determined by microdilution in 96-well plates. The ethanolic extract of the leaves of C. argyrophylloides manifested antioxidant action in the quantitative DPPH test with a significant bioactivity of 84.70 AAO% in 500 µg/mL, with an EC50 of 236.79. The content of total phenolic compounds was 946.06 mg of gallic acid equivalents/g of sample, and total flavonoids was 58.11 mg of quercetin equivalents/g of sample, the result obtained for FRAP was 15294.44 µM Trolox/g of sample and ABTS was 718 M Trolox of sample. The prospecting of the chemical constituents of the leaves of C. argyrophylloides revealed the presence of the main compounds that manifests the antioxidant activity and it was proven by the DPPH method that there is antioxidant activity in the analyzed sample, in addition to demonstrating a significant content of phenolic compounds and total flavonoid content in the species, which corroborates the antioxidant activity of the plant sample. The leaf extracts presented growth inhibition halos of 10 and 12 mm upon Staphylococcus aureus ATCC 25923.

Resumo Croton argyrophylloides Muell. Arg., pertencente à família Euphorbiaceae, conhecida popularmente como marmeleiro prateado e sacatinga, é um vegetal do bioma caatinga comumente encontrado no Nordeste do Brasil. O presente trabalho teve como objetivo avaliar a atividade antioxidante da espécie. O estudo fitoquímico foi realizado por meio de análise qualitativa dos constituintes químicos e determinação quantitativa do teor de fenóis totais pelo teste de Folin-Ciocalteu. Os testes antioxidantes qualitativos e quantitativos foram realizados pelo método do DPPH (2,2 difenil-1- picril-hidrazila) e redução do ferro (FRAP). A concentração inibitória mínima (CIM) foi determinada por microdiluição em placas de 96 poços. O extrato etanólico das folhas de C. argyrophylloides apresentou ação antioxidante no teste DPPH quantitativo com uma significativa bioatividade de 84.70 AAO% em 500 µg/mL, apresentando um CE50 de 236.79. O teor de compostos fenólicos totais, foi de 946,06 mg equivalentes de ácido gálico/g de amostra, e de flavonoides totais de 58,11 mg equivalentes de quercetina/g da amostra, o valor encontrado para FRAP foi de 15294,44 µM Trolox/g da amostra e de ABTS foi 718 M Trolox da amostra. A prospecção dos constituintes químicos das folhas de C. argyrophylloides revelou a presença dos principais compostos que caracterizam a atividade antioxidante e foi possível comprovar pelo método de DPPH que há atividade antioxidante na amostra analisada, além de demonstrar um resultado significativo de teor de compostos fenólicos e teor de flavonoides totais na espécie e o que corrobora com a atividade antioxidante da amostra vegetal. Os extratos das folhas apresentaram halos de inibição de crescimento de 10 e 12mm frente a Staphylococcus aureus ATCC 25923.

In vitro susceptibility of Sporothrix spp. to complexes coordinated with Co(II) and cobalt chloride hexahydrate

Abstract Sporothrix spp. are the major dimorphic fungus associated with a type of subcutaneous mycosis, sporotrichosis. The limitation of antifungal availability and the past reports of in vitro resistance of Sporothrix spp. clinical isolates makes it important to search for new compounds with antifungal activities. In this study, we therefore evaluate the in vitro activities of complexes coordinated with Co(II) and cobalt chloride hexahydrate against clinical isolates of Sporothrix spp. Broth microdilution test was performed as per M38-A2 from CLSI (2008) in duplicate for 31 clinical isolates of Sporothrix spp. (27 S. brasiliensis e 04 S. schenckii stricto sensu). The antifungal activities of the complexes coordinated with Co(II) and cobalt chloride hexahydrate were detected at a concentration range of 32-128 µg/mL for all isolates. None of the compounds demonstrated any cytotoxicity (to macrophage cells) at the concentration of 200 µg/mL. The activity against Sporothrix spp. recorded in this study instigate the continuity of experimental studies with Co(II) to search for the mechanisms of antifungal action as well as to evaluate its interaction with the commercial antifungal drugs.

Construction of EF-G knockdown strain of Mycobacterium smegmatis and drug resistance analysis / 生物工程学报

As the only translational factor that plays a critical role in two translational processes (elongation and ribosome regeneration), GTPase elongation factor G (EF-G) is a potential target for antimicrobial agents. Both Mycobacterium smegmatis and Mycobacterium tuberculosis have two EF-G homologous coding genes, MsmEFG1 (MSMEG_1400) and MsmEFG2 (MSMEG_6535), fusA1 (Rv0684) and fusA2 (Rv0120c), respectively. MsmEFG1 (MSMEG_1400) and fusA1 (Rv0684) were identified as essential genes for bacterial growth by gene mutation library and bioinformatic analysis. To investigate the biological function and characteristics of EF-G in mycobacterium, two induced EF-G knockdown strains (Msm-ΔEFG1(KD) and Msm-ΔEFG2(KD)) from Mycobacterium smegmatis were constructed by clustered regularly interspaced short palindromic repeats interference (CRISPRi) technique. EF-G2 knockdown had no effect on bacterial growth, while EF-G1 knockdown significantly retarded the growth of mycobacterium, weakened the film-forming ability, changed the colony morphology, and increased the length of mycobacterium. It was speculated that EF-G might be involved in the division of bacteria. Minimal inhibitory concentration assay showed that inhibition of EF-G1 expression enhanced the sensitivity of mycobacterium to rifampicin, isoniazid, erythromycin, fucidic acid, capreomycin and other antibacterial agents, suggesting that EF-G1 might be a potential target for screening anti-tuberculosis drugs in the future.

Impact of minimal inhibitory concentration breakpoint of amoxicillin and clarithromycin on the efficacy of these containing regimens for Helicobacter pylori eradication / 中华消化杂志

Objective:To evaluate the impact of amoxicillin and clarithromycin resistance on the eradication rate of Helicobacter pylori ( H. pylori), and to explore the optimal minimal inhibitory concentration (MIC) breakpoint of amoxicillin and clarithromycin. Methods:From March 2008 to December 2010, patients with H. pylori positive received standard triple therapy to eradicate H. pylori were retrospectively analyzed, 140 patients with H. pylori infetion were included, of which 12 patients did not receive eradication treatment. At 8 to 12 weeks after treatment, the eradication rate of H. pylori of 140 and 128 patients was calculated by intention-to-treat (ITT) and per-protocol population (PP) analysis, respectively. The correlation between amoxicillin and clarithromycin resistance and failure of H. pylori eradication was analyzed. And the relation between different MIC breakpoints of amoxicillin and clarithromycin and failure of H. pylori eradication was also analyzed. Binary logistic regression analysis and consistency test were used for statistical analysis. Results:The results of ITT and PP analysis indicated that the eradication rate of H. pylori of the standard triple therapy was 66.4%(93/140)and 72.7% (93/128), respectively, 95% confidence interval ( CI) 59.3% to 74.3%, and 65.6% to 79.7%, respectively. The results of binary logistic regression analysis showed that amoxicillin resistance (odds ratio ( OR)=6.326, 95% CI 1.090 to 36.725, P=0.040) and clarithromycin resistance ( OR=10.686, 95% CI 4.031 to 28.326, P<0.01) were both independent risk factors of H. pylori eradication failure. The results of consistency test demonstrated that when the MIC breakpoint of amoxicillin was 0.125 mg/L, the correlation between amoxicillin resistance and H. pylori eradication failure was the highest (fair consistency, P<0.05); when the MIC breakpoint of clarithromycin was 2.000 mg/L, the correlation between clarithromycin resistance and H. pylori eradication failure was the highest (moderate consistency, P<0.05). Conclusions:The eradication rate of H. pylori of standard triple therapy dropped to <80%. The decrease of H. pylori eradication rate was related to the resistance of amoxicillin and clarithromycin. The best MIC breakpoints of amoxicillin and clarithromycin were 0.125 and 2.000 mg/L, respectively.

Study on the antifungal effect of amphotericin B combined with fluconazole at different time points / 口腔疾病防治

Objective@#To investigate the in vitro interaction of amphotericin B (AmB) and fluconazole (FLC) at different time points and provide a reference for clinical combined treatment therapy of polyenes and azoles.@*Methods@#Candida albicans ATCC SC5314 was used in the study. The minimum inhibitory concentration (MIC) of antifungal drugs was determined using the double microdilution broth method. The same amount of DMSO and low concentration drugs were added to the DMSO treatment group at different time points (0, 2, 4, 6 h) to determine whether the solvent background environment affected the growth of Candida albicans. In the experimental group, to observe the effect of low concentration AmB on the antifungal effect of FLC, the experimental group was administered a low concentration of AmB (0.25 μg/mL or 0.125 μg/mL) added to FLC at different time points (0, 2, 4, 6 h), and the same amount of DMSO was added to FLC at different time points in the single drug control group. In the experimental group, to observe the effect of low concentration of FLC on the antifungal effect of AmB, the experimental group was administered a low concentration of FLC (0.06 μg/mL or 0.03 μg/mL) in AmB at different time points (0, 2, 4, 6 h), and the same amount of DMSO was used at different time points as the single drug control group. In the solvent group, the same amounts of DMSO and low concentration drugs were added at different time points. After resuscitation, the colony growth of each solvent control group, single-drug control group and experimental group was observed to evaluate the interaction between drug concentration and time. Compared with the AmB single-drug control group, there was no significant change in the experimental group with added low concentrations of FLC at 0 h (F=0.27, P=0.775), which was 1.74-1.93 times that of the control group at 2-4 h (P < 0.001), and there was no significant difference in colony count after 6 h (P > 0.05). @*Results@# Under the treatment of FLC at an inhibitory concentration (0.25 μg/ml), adding low concentration AMB did not affect the antifungal effect of FLC, and the multiple of colony count differences were not significant (P > 0.05).@*Conclusion@#The interaction between AmB and FLC was time-dependent. At the early stage (0 h), the interaction effect between fluconazole and amphotericin B was not clear. The fungicidal effect of AmB could be weakened when FLC was supplied at 2-4 h, and the effect of FLC on AmB was absent after 6 h.

Study on Sub-minimal Inhibitory Concentration of Tanreqing Injection in Inhibiting Hemolytic Activity of Staphylococcus aureus α-hemolysin / 中国实验方剂学杂志

Objective:To study whether Tanreqing injection （TRQ） can alleviate the body injury in the process of infection by inhibiting the production and release of <italic>α</italic>-hemolysin of <italic>Staphylococcus aureus</italic> under sub-minimal inhibitory concentration， and to provide experimental basis for better guidance of clinical medication. Method:The effects of TRQ on the minimum inhibitory concentration （MIC） and bacterial growth of <italic>S.aureus</italic> were determined firstly by microplate method and time-growth curve. The different sub-minimal inhibitory concentrations of TRQ were co-cultured with bacteria or bacterial supernatants， and then co-incubated with defibrillated rabbit blood to detect the inhibitory and neutralizing effects of TRQ on <italic>S.aureus</italic> <italic>α</italic>-hemolysin. Cell counting kit-8 （CCK-8） cell viability assay was used to detect the protective effect of TRQ on <italic>S. aureus</italic>-mediated damage to human alveolar epithelial cells （A549）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the effect of sub-minimal inhibitory concentration of TRQ on the mRNA expression of <italic>S.aureus</italic> <italic>α</italic>-hemolysin regulatory genes hla and agrA. Result:The MIC of TRQ to <italic>S.aureus </italic>was 1/8 of the stock solution， and the sub-minimal inhibitory concentration （1/64MIC-1/16MIC） TRQ used in this study did not affect the growth of bacteria. 1/64MIC-1/16 MIC TRQ had the effect of inhibiting and neutralizing the hemolytic activity of <italic>α</italic>-hemolysin， with a protective effect on <italic>S.aureus</italic> supernatant-mediated A549 cell damage， and its inhibitory effect on <italic>α</italic>-hemolysin was closely related to the inhibition of hla and agrA mRNA expression. Conclusion:The sub-minimal inhibitory concentration TRQ can inhibit and neutralize the hemolytic activity of <italic>α</italic>-hemolysin of <italic>S.aureus</italic>， with a protective effect on A549 cell damage mediated by <italic>S.aureus</italic> infection， and its mechanism of inhibiting <italic>α</italic>-hemolysin is closely related to the interference with agr regulatory system.

Emergence of multidrug-resistant bacteria isolated from surgical site infection in dogs and cats / [Emergência de bactérias multirresistentes isoladas da infecção no sítio cirúrgico em cães e gatos]

Surgical site infections (SSIs) and antimicrobial resistance among pathogens causing SSI are a growing concern in veterinary hospitals. One major reason, the widespread use of antimicrobials, has led to increased incidence of SSIs. This study identified bacteria and resistance profiles to antimicrobials in the SSI cases diagnosed at the Surgical Clinic of Small Animals in the Veterinary Hospital, Federal University of Viçosa, Brazil. The main genus identified was Staphylococcus, followed by Escherichia, Enterococcus, Bacillus, Shigella, Citrobacter, Proteus, Morganella, Serratia, Enterobacter, Pseudomonas and Klebsiella were also found, but in small number. The results indicated the predominance of Gram-negative bacteria among the collected samples. Most of isolates identified were resistant to more than one of the following antimicrobials: ampicillin, tetracycline, enrofloxacin, amoxicillin/clavulanic acid and cephalotin. Of the 17 Staphylococcus sp. isolates, two (11.8%) were methicillin-resistant Staphylococcus aureus (MRSA) and 11 (64.7%) of them were methicillin-resistant Staphylococcus pseudintermedius (MRSP). There were bacterial genera identified with resistance to all tested antimicrobials in different proportions. This should alert veterinary hospitals to the emergence of multidrug-resistant bacteria and to the requirement for the revision of surgical protocols with regard to antimicrobial prophylaxis and therapy.(AU)

As infecções em sítio cirúrgico (ISCs) e a resistência bacteriana entre os patógenos relacionados constituem uma preocupação crescente nos hospitais veterinários. O aumento na incidência de ISCs possui forte relação com o uso amplo e disseminado de antibióticos. O presente estudo identificou bactérias e perfis de resistência a antibióticos nos casos de ISCs diagnosticados na Clínica Cirúrgica de Pequenos Animais do Hospital Veterinário da Universidade Federal de Viçosa, Brasil. O principal gênero identificado foi Staphylococcus, seguido pelos gêneros Escherichia, Enterococcus, Bacillus, Shigella, Citrobacter, Proteus, Morganella, Serratia, Enterobacter, Pseudomonas e Klebsiella, porém, em menor quantidade. Os resultados demonstraram a predominância de bactérias Gram-negativas entre as amostras coletadas. A maioria dos isolados identificados eram resistentes a um ou a mais de um dos seguintes antibióticos: ampicilina, tetraciclina, enrofloxacina, amoxicilina/ácido clavulânico e cefalotina. Entre os 17 isolados de Staphylococcus sp., dois (11,8%) eram Staphylococcus aureus resistentes à meticilina (SARM) e 11 (64,7%) eram Staphylococcus pseudintermedius resistentes à meticilina (SPRM). Houve identificação de gêneros bacterianos com diferentes proporções de resistência para todos os antibióticos avaliados. Esses achados devem alertar os hospitais veterinários para a emergência de bactérias multirresistentes e para a necessidade de revisar a profilaxia e a terapia antimicrobiana referente aos protocolos cirúrgicos.(AU)

Antimicrobial activities of chlorogenic acid and 13 main metabolites in vitro / 中草药

Objective: To compare with the antimicrobial activities of chlorogenic acid and its 13 main metabolites in vivo. Methods: Main metabolites were detected by broth microdilution method against clinical isolated strains including methicillin-sensitive Staphylococcus aureus (MSSA), methicillin-resistant Staphylococcus aureus (MRSA), non ESBLs-producting Escherichia coli (E-), extended-spectrum β-lactamase Escherichia coli (E+), Klebsiella pneumoniae (E-/E+), Acinetobacter baumannii, and Pseudomonas aeruginosa to compare with the minimal inhibitory concentrations value of chlorogenic acid. Results: For the most tested strains, the antibacterial activities of some major metabolites were enhanced, ranging from four to 66 times higher than chlorogenic acid. Benzoic acid, p-hydroxybenzoic acid, p-coumaric acid, dihydrocaffeic acid, and gallic acid were stronger than the other metabolites. Moreover, the antibacterial actions of dihydrocaffeic acid and gallic acid against MSSA and MRSA was increased 33-66 times respectively. Conclusion: Some main metabolites presented more potent antibacterial activities than chlorogenic acid, and chlorogenic acid may exert antibacterial effects through some of its metabolites.

Study on Inhibitory Activity of Dracocephalum moldavica Extracts to Clinical Pathogenic Bacteria and Its Bioin- formatics / 中国药房

OBJECTIVE：To study inhibitory activity of different extracts of Dracocephalum moldavica to clinical pathogenic bacteria，and to excavate its possible antibacterial mechanism. METHODS ：After extraction by 65％ ethanol and extraction by petroleum ether ，dichloromethane，ethyl acetate ，n-butanol，the different polar fractions of D. moldavica were obtained . Taking Klebsiella pneumoniae ，Staphylococcus aureus and other clinical multiple resistant pathogens as objects，the diameter of inhibition zone of different extraction fractions was measured by paper diffusion method ，and the antibacterial active fraction was screened. The minimal inhibitory concentration （MIC）of antibacterial active fraction to common clinical pathogens was determined by agar dilution method ；the growth curve of MRSA was drawn by turbidimetric method. The differentially expressed protein between antibacterial active fraction group and control group was screened by PEAK ® Q 8.5 software，and the gene ontology （GO）analysis and KEGG signaling pathway enrichment analysis were carried out by Blast 2 GO and KOBAS 3.0 online software. RESULTS ： Petroleum ether ，dichloromethane，ethyl acetate and n-butanol fraction of D. moldavica had no significant inhibitory effect on Gram-negative bacteria. The ethyl acetate fraction of D. moldavica had antibacterial activity in varying degrees against several kinds of Gram-positive bacteria as Staphylococcus aureus ，S. epidermidis；the diameter of inhibition zone was 10-16 mm，which was the active fraction. MICs of ethyl acetate fraction to S. aureus ，S. epidermidis and S. hominis were all 0.781 3 mg/mL；MIC to S. saprophyticus was 0.390 7 mg/mL；MICs to S. saprophyticus and standard strain of S. aureus were both 1.562 5 mg/mL. The 1， 2 times MIC of ethyl acetate could inhibit the growth of MRSA ，and the inhibitory activity increased with the increase of dose. A total of 300 differentially expressed proteins were screened （P＜0.01），of which 239 were up-regulated and 61 were down-regulated. The differentially expressed proteins were （No.81260478，No.816- mainly concentrated in cell sites such as cells ，cellular com- 60048） ponent，etc.，and in metabolic process such as cell process ， biological process and molecular functions such as catalytic activity，protein binding ，etc. They were mainly concentratedwang.zhanli@hotmail.com in microbial metabolism in different environments ，fructose and mannose metabolism signaling pathway （P＜0.05）. CONCLUSIONS ：The ethyl acetate fraction of D. Moldavica is the antibacterial active fraction ，and its activity may be related to the microbial metabolism and cell glycometablism.

Antimicrobial efficacy of novel ethanolic extract of morinda citrifolia against enterococcus faecalis by agar well diffusion and broth dilution methods - an invitro study / Eficácia de um novo antimicrobiano etanólico de extrato de Moringa citrifolia contra Enterococcus Faecalis por meio dos métodos de difusão em ágar e de diluição em caldo ­ um estudo in vitro

Objective: The long-term success of root canal treatment is ultimately related to the effective debridement and disinfection of the root canal system. Hence, the irrigants play an important role in achieving the good penetrability and bactericidal activity. The present study was mainly aimed at evaluating the invitro antimicrobial efficacy of Novel Ethanolic Extract of Morinda Citrifolia by agar well diffusion and broth dilution methods. Material and methods: The antibacterial effect of Ethanolic Extract of Morinda Citrifolia was investigated against Enterococcus Faecalis (E. Faecalis). Agar well diffusion and broth dilution methods were used to determine the Minimal Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) Results: The MIC of Ethanolic Extract of Morinda Citrifolia extract was found to be 12.5 mg/ml and the MBC was found to be 25 mg/ml Conclusion: Novel Ethanolic Extract of Morinda Citrifolia possess antimicrobial activity against E.Faecalis. But still, future studies are needed. (AU)

Objetivo: O sucesso a longo prazo do tratamento de canais radiculares está intimamente relacionada com a efi cácia do debridamento e desinfecção do Sistema do canal radicular. Consequentemente, os irrigantes têm um importante papel na capacidade de penetração e atividade bactericida. O presente estudo teve como objetivo principal a avaliação in vitro da efi cácia antimicrobiana do novo extrato etanólico de Morinda Citrifolia por meio dos métodos de difusão em ágar ou de diluição em caldo. Material e métodos: O efeito antibacteriano do extrato etanólico de Morinda Citrifolia foi investigado contra Enterococcus Faecalis (E. Faecalis). Os métodos de diluição em ágar e de diluição em caldo foram usados para determinar a concentração inibitória minima (MIC) e concentração bactericida minima (MBC). Resultados: O MIC do extrato etanólico de Morinda Citrifolia foi obtido na concentração de 12.5mg/ml e a MBC foi de 25mg/ml Conclusão: O novo extrato etanólico de Morinda Citrifolia apresenta atividade antimicrobiana contra E.Faecalis. Porém, futuros estudos são necessários. (AU)

In vitro combination antimicrobial susceptibility of fosfomycin sodium / 中国感染控制杂志

Objective To evaluate in vitro antimicrobial effect of fosfomycin sodium single use and combination with other antimicrobial agents on clinically isolated Staphylococcus aureus (S.aureus), Klebsiella pneumoniae (K.pneumoniae) and Pseudomonas aeruginosa (P.aeruginosa) in China.Methods Combined antimicrobial susceptibility testing was performed with checkerboard method, minimal inhibitory concentrations (MICs) were detected by two-fold agar dilution method, susceptibility of S.aureus (n=113 strains), K.pneumoniae (n=108 strains), and P.aeruginosa (n=110 strains) isolated from 18 hospitals in China in recent three years was determined by single and combined antimicrobial susceptibility testing.Results MIC50 value of fosfomycin sodium single use were all≤32 mg/L against all tested strains, regardless of whether strains were resistant to other antimicrobial agents or not.The synergistic rate of fosfomycin sodium with levofloxacin, minocycline, oxacillin, and clindamycin against methicillin-resistant S.aureus (MRSA) was＞43％.Synergistic rate of fosfomycin sodium with levofloxacin and imipenem against imipenem-nonsusceptible P.aeruginosa was＞35％, synergistic rates of fosfomycin sodium with tested antimicrobial agents against imipenem-susceptible P.aeruginosa were all＞35％.Conclusion Fosfomycin sodium still has good antimicrobial activity against common clinical drug-resistant bacteria, such as MRSA, extended-spectrumβ-lactamase-producing K.pneumoniae and so on, it has synergistic effect with many other kinds of antimicrobial agents, suggesting that in the limited treatment of infection caused by drug-resistant bacteria, fosfomycin in combination with other antimicrobial agents may be a useful choice.

In vitro analysis of the minimal inhibitory concentration values of different generations of anti-methicillin-resistant Staphylococcus aureus antibiotics

Methicillin-resistant Staphylococcus aureus (MRSA) resistance to antimicrobials may result in the increased risk of treatment failure. The objective of the study was to analyse in vitro MRSA susceptibility to vancomycin, linezolid, daptomycin, tigecycline, ceftaroline, dalbavancin, clindamycin, ciprofloxacin and trimethoprim/sulfamethoxazole. All MRSA strains isolated from hospitalised patients were analysed according to the current microbiological recommendations. Finally, a total of 124 MRSA strains were analysed; all were susceptible to tested antibiotics. Dalbavancin had the lowest minimum inhibitory concentration (MIC), and vancomycin the highest MIC value. There were 28/124 strains of MRSA susceptible for clindamycin, 36/124 for ciprofloxacin and 121/124 for trimethoprim/sulfamethoxazole. Dalbavancin was the most effective antimicrobial in our study.

Phytochemistry and antibacterial activity of aqueous andhydroalcoholic extracts of three medicinal plants against foodpathogens / Fitoquímica e atividade antibacteriana de extratos aquosos e hidroalcoólicos de trêsplantas medicinais contra patógenos de alimentos

This study aimed to characterize the phytochemical compositions of three medicinal Brazilian plants’ leaves and bast extracts, and to determine their antibacterial activity on three foodborne and waterborne bacterial pathogens. Parkia platycephala, Pouteria ramiflora and Lophanthera lactescens leaves and basts were collected and aqueous and hydroalcoholic extracts were prepared. Qualitative screening of the phytochemical extracts was performed with three replicates and in triplicate in order to identify the bioactive compounds. The Minimal Inhibitory Concentration and Minimal Bactericide Concentration were determined by microdilution in broth and Escherichia coli, Salmonella typhimurium and Staphylococcus aureus growth was observed on agar plates. Phytochemical composition analysis allowed for the identification of anthraquinones, catechins, saponins, tannins, sesquiterpenlactones and other lactones in the three plants’ leaves and bast aqueous and hydroalcoholic extracts. Eighty-three percent of the plant extracts showed antibacterial activity against S. aureus, and P. platycephala extracts were the only ones that inhibited E. coli and S. typhimurium growth. The present study contributes significantly to the phytochemical composition characterization of three plant species commonly used in Brazilian traditional medicine. The plant extracts’ in vitro antibacterial activity was demonstrated and catechins present in the extracts are, most likely, the bioactive compounds responsible for this action.

Este estudo objetivou caracterizar a composição fitoquímica dos extratos de folhas e das entrecascas de três plantas medicinais brasileiras e determinar a sua atividade antimicrobiana contra três patógenos bacterianos de alimentos. Foram elaborados extratos aquosos e hidroalcoólicos, por meio de folhas e de entrecascas de Parkiaplatycephala, Pouteriaramiflora e Lophantheralactescens. O estudo qualitativo dos extratos foi realizado com três réplicas, em triplicata, para permitir a identificação dos compostos bioativos. A Concentração Inibitória Mínima e a Concentração Bactericida Mínima foram determinadas por microdiluição contra Escherichia coli, Salmonella typhimurium e Staphylococcus aureus. A análise da composição fitoquímica permitiu identificar antraquinonas, catequinas, saponinas, taninas, sesquiterpenlactonas e outras lactonas nos extratos aquosos e hidroalcoólicos das folhas e das entrecascas das três plantas. Oitenta e três porcento dos extratos das plantas apresentaram atividade antibacteriana contra S. aureus. Os extratos de P. platycephala foram os únicos que inibiram o crescimento de E. coli e S. typhimurium. Este estudo contribui significativamente para a caracterização da composição fitoquímica de três espécies de plantas, frequentemente, utilizadas na medicina tradicional brasileira. A atividade antibacteriana, in vitro, dos extratos das plantas foi demonstrada, e as catequinas são, provavelmente, o composto bioativo responsável por essa atividade.

Inhibitory effects of nano silver and titanium dioxide on the nucleic acid of staphylococcus aureus / 中华急诊医学杂志

Objective To study the effects of nano silver (Ag) and titanium dioxide (TiO2) on the content of nucleic acid in staphylococcus aureus in order to explore their antibacterial mechanisms.Methods After preparation of beef extract peptone liquid cultures,the effects of minimal inhibitory concentrations (MICs) of nano Ag and TiO2 on staphylococcus aureus strains were determined.With the 1/2 MICs nano Ag and TiO2,the contents of DNA and RNA macromolecules from staphylococcus aureus cultures were measured to determine the damage degree of staphylococcus aureus cell membranes by ultraviolet spectrophotometer,and then the fluorescence intensities of the staphylococcus aureus cells were observed under fluorescence microscope and the fluorescence values were tested by fluorescence spectrophotometer to determine the contents of nucleic acid DNA and RNA.Results The MICs of nano Ag and TiO2 were 1.6 mg/mL and 5.781 μg/mL.After treatment with the 1/2 MICs nano Ag and TiO2,nano Ag group and TiO2 group were compared with the control group (culture fluid without adding antibacterial agent),respectively,and there were no significant differences in the contents of DNA and RNA macromolecules from staphylococcus aureus cultures between n anoAg group and control group as well as between TiO2 group and control group were (P＞0.05),and there were significant decreases in fluorescence intensities and the contents of nucleic acid DNA and RNA (P＜0.01).Conclusions Nano Ag and TiO2 had obvious antibacterial effects on staphylococcus aureus and the antibacterial properties of nano Ag was stronger than that of TiO2.The antibacterial mechanisms of nano Ag and TiO2 against staphylococcus aureus may be associated with the inhibition of the synthesis of nucleic acid DNA and RNA,inhibiting protein synthesis and then bacterial growth.

Effect of sub-minimal inhibitory concentration piperacillin/tazobactam on biofilm formation of Escherichia coli isolates / 重庆医学

Objective To study the effect of sub-minimal inhibitory concentration(sub-MIC) piperacillin/tazobactam on the biofilm formation of Escherichia coli to provide the theoretical basis for clinical anti-infection treatment.Methods The minimal inhibitory concentration of E.coli was detected by broth microdilution method.The biofilm formation ability was analyzed by using the 96-well crystal violet staining method.The bacterial adhesion ability was determined by adopting the flat colony counting method.The effect of sub-MIC piperacillin/tazobactam on the motility of E.coli was determined by adopting the swimming movement.Results Sub-MIC piperacillin/tazobactam could significantly inhibit the biofilm formation of E.coli(P＜ 0.05).In addition,subMIC piperacillin/tazobactam not only significantly reduced bacterial adhesion,but also decreased the bacterial motility.Conclusion Sub-MIC piperacillin/tazobactam might inhibit the biofilm formation of E.coli by inhibiting its adhesion and motility.