Spectral analysis, in vitro cytotoxicity and antibacterial studies of bioactive principles from the leaves of Conocarpus lancifolius, a common tree of Jazan, Saudi Arabia

Abstract The objective of the present study was to analyse the bioactive compounds of the leaves of Conocarpus lancifolius (C. lancifolius). The GC-MS analysis of the hot methanolic extract of the leaves (HMEL) of C. lancifolius exhibited the bioactive compounds such as 1-(3-Methoxy-2-nitrobenzyl) iso quinoline, morphin-4-ol-6,7-dione, 1-bromo-N-methyl-, phytol, hexadecanoic acid, 2,3-dihydroxypropyl ester, 2,2':4',2-terthiophene, ethyl iso-allocholate, caryophyllene oxide, campesterol, epiglobulol, cholestan-3-ol, 2-methylene-, (3á,5à)-, dasycarpidan-1-methanol, acetate (ester) and oleic acid, eicosyl ester. The FT-IR analysis of HMEL of C. lancifolius showed a unique peak at 3184, 2413, 1657 cm-1 representing coumaric acid, chlorogenic acid and ferulic acid. The HMEL of C. lancifolius was actively inhibiting the proliferation of breast cancer cells MCF-7 ATCC at the concentration of 72.66 ± 8.21 µg/ml as IC50 value. The HMEL of C. lancifolius also revealed a good spectrum of activity against Gram-positive and Gram-negative bacterial cultures screened in this work. The activity observed has shown more or less similar effects against screened bacteria. However, the magnitude of potentiality was significantly lesser compared to standard ciprofloxacin disc at p 0.001 level (99% confidence intervals). Furthermore, the study demonstrating the bioactive compounds can be isolated from the leaves of C. lancifolius.

Resumo O objetivo do presente estudo foi analisar os compostos bioativos das folhas de Conocarpus lancifolius (C. lancifolius). A análise por GC-MS do extrato metanólico quente das folhas (HMEL) de C. lancifolius exibiu os compostos bioativos como 1- (3-Metoxi-2-nitrobenzil) isoquinolina, morfina-4-ol-6,7- diona, 1-bromo-N-metil-, fitol, ácido hexadecanoico, 2,3-di-hidroxipropil éster, 2,2 ': 4', 2 - tertiofeno, isoalocolato de etil, óxido de cariofileno, campesterol, epiglobulol, colestano -3-ol, 2-metileno-, (3á, 5à) -, dasycarpidan-1-metanol, acetato (éster) e ácido oleico, éster eicosílico. A análise FT-IR de HMEL de C. lancifolius mostrou um pico único em 3184, 2413, 1657 cm-1 representando ácido cumarico, ácido clorogênico e ácido ferúlico. O HMEL de C. lancifolius inibiu ativamente a proliferação de células de câncer de mama MCF-7 ATCC na concentração de 72,66 ± 8,21 µg / ml como valor de IC50. O HMEL de C. lancifolius também revelou bom espectro de atividade contra culturas de bactérias Gram-positivas e Gram-negativas rastreadas neste trabalho. A atividade observada mostrou efeitos mais ou menos semelhantes contra bactérias rastreadas. No entanto, a magnitude da potencialidade foi significativamente menor em comparação com o disco de ciprofloxacina padrão em nível de p 0,001 (intervalos de confiança de 99%). Além disso, o estudo demonstrando os compostos bioativos pode ser isolado das folhas de C. lancifolius.

Antioxidant and antimicrobial activities of Thymus vulgaris essential oil contained and synthesis thymus (Vulgaris) silver nanoparticles

Abstract Several species of thymus have therapeutic properties, so they are used in traditional medicine. In this work was carried out to synthesize Thymus vulgalis silver nanoparticles (TSNPS) and evaluate antioxidant and antimicrobial activities of TSNPS and T. vulgalis essential oil extract (TEOE). The essential oils analyzed by GC-MS and were characterized. Major compounds of phenol, 2 methyl 5 (1 methylethyle) (CAS), thymol and 1,2 Benzene dicarboxylic acid, 3 nitro (CAS) (48.75%, 32.42% and 8.12%, respectively) were detected. Results demonstrated that the TSNPS gave a highest DPPH radical scavenging activity, it was obtained 97.2 at 1000 ug/ml. TSNPS, Thymus + Hexane (T+H), Thymus + Ethanol (T+E) gave the greatest antimicrobial activity than amoxicillin (AM) and ciprofloxacin (CIP). In conclusion: The essential oil of thymus (Vulgaris) and thymus (Vulgaris) silver nanoparticles can be a good source of natural preservatives as an antioxidant and antimicrobial agents for increasing the shelf life of foodstuffs.

Resumo Diversas espécies de timo possuem propriedades terapêuticas, por isso são utilizadas na medicina tradicional. Neste trabalho foi realizado para sintetizar nanopartículas de prata Thymus vulgalis (TSNPS) e avaliar as atividades antioxidante e antimicrobiana de TSNPS e extrato de óleo essencial de T. vulgalis (TEOE). Os óleos essenciais analisados por GC-MS e foram caracterizados. Os principais compostos de fenol, 2 metil 5 (1 metiletilo) (CAS), timol e ácido 1,2 Benzenodicarboxílico, 3 nitro (CAS) (48,75%, 32,42% e 8,12%, respectivamente) foram detectados. Os resultados demonstraram que o TSNPS deu uma maior atividade de eliminação do radical DPPH , foi obtido 97,2 a 1000 ug / ml. TSNPS, Timo + Hexano (T + H), Timo + Etanol (T + E) deu a maior atividade antimicrobiana do que amoxicilina (AM) e ciprofloxacina (CIP). Em conclusão: O óleo essencial de nanopartículas de prata do timo (Vulgaris) e do timo (Vulgaris) pode ser uma boa fonte de conservantes naturais como agentes antioxidantes e antimicrobianos para aumentar a vida útil de alimentos.

Eugenia uniflora: a promising natural alternative against multidrug-resistant bacteria / Eugenia uniflora: uma alternativa natural promissora contra bactérias multirresistentes

This work aimed to evaluate the chemical composition, antioxidant and antimicrobial activities from crude extract and fractions from leaves of Eugenia uniflora Linn. The crude extract was obtained by turbo extraction and their fractions by partitioning. Chromatographic analysis were performed, and the antioxidant capacity was verified by two methods (DPPH• and ABTS•+). The Minimal Inhibitory/Bactericidal Concentration were conducted against twenty-two bacteria, selecting five strains susceptible to extract/fractions and resistant to the antibiotics tested. Ampicillin, azithromycin, ciprofloxacin, and gentamicin were associated with Ethyl Acetate Fraction (EAF) against multidrug-resistant strains in modulatory and checkerboard tests. The chromatographic data showed gallic acid, ellagic acid, and myricitrin in crude extract, with enrichment in the EAF. The electron transfer activity demonstrated in the antioxidant tests is related to the presence of flavonoids. The Gram-positive strains were more susceptible to EAF, and their action spectra were improved by association, comprising Gram-negative bacilli. Synergisms were observed to ciprofloxacin and gentamicin against Pseudomonas aeruginosa colistin-resistant. The results demonstrate that the extract and enriched fraction obtained from the leaves of E. uniflora act as a promising natural alternative against multidrug-resistant bacteria.

Este trabalho teve como objetivo avaliar a composição química, as atividades antioxidantes e antimicrobianas do extrato bruto e frações de folhas de Eugenia uniflora Linn. O extrato bruto foi obtido por turbólise e suas frações por partição. Foram realizadas análises cromatográficas e a capacidade antioxidante foi verificada por dois métodos (DPPH• e ABTS•+). A Concentração Inibitória/Bactericida Mínima foi realizada contra vinte e duas bactérias, selecionando cinco cepas suscetíveis a extração/frações e resistentes aos antibióticos testados. Ampicilina, azitromicina, ciprofloxacina e gentamicina foram associados à Fração Acetato de Etila (FAE) contra cepas multirresistentes em testes modulatórios e de checkboard. Os dados cromatográficos mostraram ácido gálico, ácido elágico e miricitrina em extrato bruto, com enriquecimento na FAE. A atividade de transferência de elétrons demonstrada nos testes antioxidantes está relacionada com a presença de flavonoides. As cepas de Gram-positivas foram mais suscetíveis à FAE, e seus espectros de ação foram melhorados por associação, compreendendo bacilos Gram-negativos. Foram observados sinergismos de ciprofloxacina e gentamicina contra Pseudomonas aeruginosa resistente à colistina. Os resultados demonstram que o extrato e a fração enriquecida obtida das folhas de E. uniflora atuam como uma alternativa natural promissora contra bactérias multirresistentes.

Aeromonassp. in freshwater fish and antimicrobial resistance: emerging pathogen / Quantificação deAeromonassp. em peixes de água doce e resistência a antimicrobianos: patógeno emergente

ABSTRACT: The bacteria Aeromonassp. are naturally reported in aquatic ecosystems and possess pathogenic potential, being considered as emerging pathogens in humans and animals. They also cause considerable losses in fish farming and, through water, can contaminate numerous foods. This study quantified and analyzed the antimicrobial resistance profile of Aeromonassp. in fish. A total of 72 samples of two fish varieties (leather fish, Pseudoplatystomafasciatumx Leiariusmarmoratusand round fish, Colossomamacropomumx Piaractusmesopotamicusand Colossomamacropomumx Piaractusbrachypomus) were purchased from two types of sources (fresh and frozen) and three commercial establishments (supermarket, market, and fishmonger). The 55 isolated Aeromonascultures were evaluated for their antimicrobial resistance profile by the disc diffusion method. Upon quantification, the count of Aeromonassp. ranged from 4.22 to 6.00 Log CFU/g; ten different species, including A. eucrenophila, A. hydrophila, A. caviae, A. media, A. jandaei, A. veroniibv. sobria, A. trota, A. schubertii, A. veroniibv. veronii, and A. shigelloides, were identified. Among the 55 isolates, 64.45% showed resistance to Ampicillin-sulbactam, and 75% were sensitive to gentamicin and ciprofloxacin. It was concluded that 100% of the evaluated samples were contaminated by Aeromonassp., which may present a risk to consumer health since bacteria can be etiological agents of Foodborne Diseases. The antimicrobial resistance profile showed resistance to ampicillin and multi-resistance to different classes of antimicrobials, demonstrating problems with choosing an antimicrobial for treatment of any disease.

RESUMO: Aeromonas sp. são bactérias presentes naturalmente no ecossistema aquático e apresentam potencial patogênico, sendo considerados patógenos emergentes em humanos e animais. Ainda, causam perdas consideráveis em piscicultura e, através da água, podem também contaminar inúmeros alimentos. Diante disso o objetivo do trabalho foi quantificar e analisar o perfil de resistência aos antimicrobianos de bactérias do gênero Aeromonas sp. em peixes. Um total de 72 amostras foram adquiridas, sendo duas variedades de peixes (peixe de couro Pseudoplatystoma fasciatum x Leiarius marmoratus e os peixes redondos Colossoma macropomum x Piaractus mesopotamicus e Colossoma macropomum x Piaractus brachypomus), dois tipos (fresco e congelado) e de três estabelecimentos comerciais (supermercado, feira e peixaria). As 55 culturas isoladas de Aeromonas foram avaliadas quanto ao perfil de resistência aos antimicrobianos pelo método de difusão em disco. Os valores encontrados na quantificação de Aeromonas sp. variaram de 4,22 a 6,00 Log UFC/g, e foram identificadas dez espécies diferentes, sendo A. eucrenophila, A. hydrophila, A. caviae, A. media, A. jandaei, A. veronii bv. sobria, A. trota, A. schubertii, A. veronii bv. veronii e A. shigelloides. Entre os 55 isolados, 64,45% apresentaram resistência a Ampicilina-sulbactam, 75% foram sensíveis à gentamicina e ciprofloxacina. Concluiu-se que 100% das amostras avaliadas estavam contaminadas por Aeromonas sp. podendo apresentar risco à saúde do consumidor visto que estas podem ser agentes etiológicos de Doenças Transmitidas por Alimentos (DTAs). O perfil de resistência aos antimicrobianos comprovou a resistência a ampicilina e apresentou multirresistência a diferentes classes de antimicrobianos, demonstrando um problema relacionado à opção de antimicrobiano para tratamento no caso do desenvolvimento de alguma doença.

Spectral analysis, in vitro cytotoxicity and antibacterial studies of bioactive principles from the leaves of Conocarpus lancifolius, a common tree of Jazan, Saudi Arabia / Análise espectral, citotoxicidade in vitro e estudos antibacterianos de princípios bioativos das folhas de Conocarpus lancifolius, uma árvore comum de Jazan, Arábia Saudita

The objective of the present study was to analyse the bioactive compounds of the leaves of Conocarpus lancifolius (C. lancifolius). The GC-MS analysis of the hot methanolic extract of the leaves (HMEL) of C. lancifolius exhibited the bioactive compounds such as 1-(3-Methoxy-2-nitrobenzyl) iso quinoline, morphin-4-ol-6,7-dione, 1-bromo N-methyl-, phytol, hexadecanoic acid, 2,3-dihydroxypropyl ester, 2,2’:4’,2”-terthiophene, ethyl iso-allocholate, caryophyllene oxide, campesterol, epiglobulol, cholestan-3-ol, 2-methylene-, (3á,5à)-, dasycarpidan-1-methanol, acetate (ester) and oleic acid, eicosyl ester. The FT-IR analysis of HMEL of C. lancifolius showed a unique peak at 3184, 2413, 1657 cm-¹ representing coumaric acid, chlorogenic acid and ferulic acid. The HMEL of C. lancifolius was actively inhibiting the proliferation of breast cancer cells MCF-7 ATCC at the concentration of 72.66 ± 8.21 µg/ml as IC50 value. The HMEL of C. lancifolius also revealed a good spectrum of activity against Gram-positive and Gram negative bacterial cultures screened in this work. The activity observed has shown more or less similar effects against screened bacteria. However, the magnitude of potentiality was significantly lesser compared to standard ciprofloxacin disc at p< 0.001 level (99% confidence intervals). Furthermore, the study demonstrating the bioactive compounds can be isolated from the leaves of C. lancifolius.

O objetivo do presente estudo foi analisar os compostos bioativos das folhas de Conocarpus lancifolius (C. lancifolius). A análise por GC-MS do extrato metanólico quente das folhas (HMEL) de C. lancifolius exibiu os compostos bioativos como 1- (3-Metoxi-2-nitrobenzil) isoquinolina, morfina-4-ol-6,7- diona, 1-bromo-N-metil-, fitol, ácido hexadecanoico, 2,3-di-hidroxipropil éster, 2,2 ‘: 4’, 2 ” - tertiofeno, isoalocolato de etil, óxido de cariofileno, campesterol, epiglobulol, colestano -3-ol, 2-metileno-, (3á, 5à) -, dasycarpidan-1-metanol, acetato (éster) e ácido oleico, éster eicosílico. A análise FT-IR de HMEL de C. lancifolius mostrou um pico único em 3184, 2413, 1657 cm-¹ representando ácido cumarico, ácido clorogênico e ácido ferúlico. O HMEL de C. lancifolius inibiu ativamente a proliferação de células de câncer de mama MCF-7 ATCC na concentração de 72,66 ± 8,21 µg/ml como valor de IC50. O HMEL de C. lancifolius também revelou bom espectro de atividade contra culturas de bactérias Gram-positivas e Gram-negativas rastreadas neste trabalho. A atividade observada mostrou efeitos mais ou menos semelhantes contra bactérias rastreadas. No entanto, a magnitude da potencialidade foi significativamente menor em comparação com o disco de ciprofloxacina padrão em nível de p < 0,001 (intervalos de confiança de 99%). Além disso, o estudo demonstrando os compostos bioativos pode ser isolado das folhas de C. lancifolius.

Antioxidant and antimicrobial activities of Thymus vulgaris essential oil contained and synthesis thymus (Vulgaris) silver nanoparticles / Atividades antioxidante e antimicrobiana do óleo essencial de Thymus vulgaris contido e síntese de nanopartículas de prata de timo (Vulgaris)

Several species of thymus have therapeutic properties, so they are used in traditional medicine. In this work was carried out to synthesize Thymus vulgalis silver nanoparticles (TSNPS) and evaluate antioxidant and antimicrobial activities of TSNPS and T. vulgalis essential oil extract (TEOE). The essential oils analyzed by GC-MS and were characterized. Major compounds of phenol, 2 methyl 5 (1 methylethyle) (CAS), thymol and 1,2 Benzene dicarboxylic acid, 3 nitro (CAS) (48.75%, 32.42% and 8.12%, respectively) were detected. Results demonstrated that the TSNPS gave a highest DPPH radical scavenging activity, it was obtained 97.2 at 1000 ug/ml. TSNPS, Thymus + Hexane (T+H), Thymus + Ethanol (T+E) gave the greatest antimicrobial activity than amoxicillin (AM) and ciprofloxacin (CIP). In conclusion: The essential oil of thymus (Vulgaris) and thymus (Vulgaris) silver nanoparticles can be a good source of natural preservatives as an antioxidant and antimicrobial agents for increasing the shelf life of foodstuffs.

Diversas espécies de timo possuem propriedades terapêuticas, por isso são utilizadas na medicina tradicional. Neste trabalho foi realizado para sintetizar nanopartículas de prata Thymus vulgalis (TSNPS) e avaliar as atividades antioxidante e antimicrobiana de TSNPS e extrato de óleo essencial de T. vulgalis (TEOE). Os óleos essenciais analisados por GC-MS e foram caracterizados. Os principais compostos de fenol, 2 metil 5 (1 metiletilo) (CAS), timol e ácido 1,2 Benzenodicarboxílico, 3 nitro (CAS) (48,75%, 32,42% e 8,12%, respectivamente) foram detectados. Os resultados demonstraram que o TSNPS deu uma maior atividade de eliminação do radical DPPH , foi obtido 97,2 a 1000 ug / ml. TSNPS, Timo + Hexano (T + H), Timo + Etanol (T + E) deu a maior atividade antimicrobiana do que amoxicilina (AM) e ciprofloxacina (CIP). Em conclusão: O óleo essencial de nanopartículas de prata do timo (Vulgaris) e do timo (Vulgaris) pode ser uma boa fonte de conservantes naturais como agentes antioxidantes e antimicrobianos para aumentar a vida útil de alimentos.

Spectral analysis, in vitro cytotoxicity and antibacterial studies of bioactive principles from the leaves of Conocarpus lancifolius, a common tree of Jazan, Saudi Arabia / Análise espectral, citotoxicidade in vitro e estudos antibacterianos de princípios bioativos das folhas de Conocarpus lancifolius, uma árvore comum de Jazan, Arábia Saudita

Abstract The objective of the present study was to analyse the bioactive compounds of the leaves of Conocarpus lancifolius (C. lancifolius). The GC-MS analysis of the hot methanolic extract of the leaves (HMEL) of C. lancifolius exhibited the bioactive compounds such as 1-(3-Methoxy-2-nitrobenzyl) iso quinoline, morphin-4-ol-6,7-dione, 1-bromo-N-methyl-, phytol, hexadecanoic acid, 2,3-dihydroxypropyl ester, 2,2':4',2"-terthiophene, ethyl iso-allocholate, caryophyllene oxide, campesterol, epiglobulol, cholestan-3-ol, 2-methylene-, (3á,5à)-, dasycarpidan-1-methanol, acetate (ester) and oleic acid, eicosyl ester. The FT-IR analysis of HMEL of C. lancifolius showed a unique peak at 3184, 2413, 1657 cm-1 representing coumaric acid, chlorogenic acid and ferulic acid. The HMEL of C. lancifolius was actively inhibiting the proliferation of breast cancer cells MCF-7 ATCC at the concentration of 72.66 ± 8.21 µg/ml as IC50 value. The HMEL of C. lancifolius also revealed a good spectrum of activity against Gram-positive and Gram-negative bacterial cultures screened in this work. The activity observed has shown more or less similar effects against screened bacteria. However, the magnitude of potentiality was significantly lesser compared to standard ciprofloxacin disc at p< 0.001 level (99% confidence intervals). Furthermore, the study demonstrating the bioactive compounds can be isolated from the leaves of C. lancifolius.

Resumo O objetivo do presente estudo foi analisar os compostos bioativos das folhas de Conocarpus lancifolius (C. lancifolius). A análise por GC-MS do extrato metanólico quente das folhas (HMEL) de C. lancifolius exibiu os compostos bioativos como 1- (3-Metoxi-2-nitrobenzil) isoquinolina, morfina-4-ol-6,7- diona, 1-bromo-N-metil-, fitol, ácido hexadecanoico, 2,3-di-hidroxipropil éster, 2,2 ': 4', 2 " - tertiofeno, isoalocolato de etil, óxido de cariofileno, campesterol, epiglobulol, colestano -3-ol, 2-metileno-, (3á, 5à) -, dasycarpidan-1-metanol, acetato (éster) e ácido oleico, éster eicosílico. A análise FT-IR de HMEL de C. lancifolius mostrou um pico único em 3184, 2413, 1657 cm-1 representando ácido cumarico, ácido clorogênico e ácido ferúlico. O HMEL de C. lancifolius inibiu ativamente a proliferação de células de câncer de mama MCF-7 ATCC na concentração de 72,66 ± 8,21 µg / ml como valor de IC50. O HMEL de C. lancifolius também revelou bom espectro de atividade contra culturas de bactérias Gram-positivas e Gram-negativas rastreadas neste trabalho. A atividade observada mostrou efeitos mais ou menos semelhantes contra bactérias rastreadas. No entanto, a magnitude da potencialidade foi significativamente menor em comparação com o disco de ciprofloxacina padrão em nível de p < 0,001 (intervalos de confiança de 99%). Além disso, o estudo demonstrando os compostos bioativos pode ser isolado das folhas de C. lancifolius.

Antioxidant and antimicrobial activities of Thymus vulgaris essential oil contained and synthesis thymus (Vulgaris) silver nanoparticles / Atividades antioxidante e antimicrobiana do óleo essencial de Thymus vulgaris contido e síntese de nanopartículas de prata de timo (Vulgaris)

Abstract Several species of thymus have therapeutic properties, so they are used in traditional medicine. In this work was carried out to synthesize Thymus vulgalis silver nanoparticles (TSNPS) and evaluate antioxidant and antimicrobial activities of TSNPS and T. vulgalis essential oil extract (TEOE). The essential oils analyzed by GC-MS and were characterized. Major compounds of phenol, 2 methyl 5 (1 methylethyle) (CAS), thymol and 1,2 Benzene dicarboxylic acid, 3 nitro (CAS) (48.75%, 32.42% and 8.12%, respectively) were detected. Results demonstrated that the TSNPS gave a highest DPPH radical scavenging activity, it was obtained 97.2 at 1000 ug/ml. TSNPS, Thymus + Hexane (T+H), Thymus + Ethanol (T+E) gave the greatest antimicrobial activity than amoxicillin (AM) and ciprofloxacin (CIP). In conclusion: The essential oil of thymus (Vulgaris) and thymus (Vulgaris) silver nanoparticles can be a good source of natural preservatives as an antioxidant and antimicrobial agents for increasing the shelf life of foodstuffs.

Resumo Diversas espécies de timo possuem propriedades terapêuticas, por isso são utilizadas na medicina tradicional. Neste trabalho foi realizado para sintetizar nanopartículas de prata Thymus vulgalis (TSNPS) e avaliar as atividades antioxidante e antimicrobiana de TSNPS e extrato de óleo essencial de T. vulgalis (TEOE). Os óleos essenciais analisados por GC-MS e foram caracterizados. Os principais compostos de fenol, 2 metil 5 (1 metiletilo) (CAS), timol e ácido 1,2 Benzenodicarboxílico, 3 nitro (CAS) (48,75%, 32,42% e 8,12%, respectivamente) foram detectados. Os resultados demonstraram que o TSNPS deu uma maior atividade de eliminação do radical DPPH , foi obtido 97,2 a 1000 ug / ml. TSNPS, Timo + Hexano (T + H), Timo + Etanol (T + E) deu a maior atividade antimicrobiana do que amoxicilina (AM) e ciprofloxacina (CIP). Em conclusão: O óleo essencial de nanopartículas de prata do timo (Vulgaris) e do timo (Vulgaris) pode ser uma boa fonte de conservantes naturais como agentes antioxidantes e antimicrobianos para aumentar a vida útil de alimentos.

Evaluation of the antimicrobial effect of the Origanum vulgare L essential oil on strains of Klebsiella pneumoniae / Avaliação do efeito antimicrobiano do óleo essencial de Origanum vulgare L. sobre cepas de Klebsiella pneumoniae

Bacteria may be the initial cause of certain pathologies as well as a secondary agent responsible for the development of complications such as pressure ulcer infections. Pressure ulcers are a persistent health problem, especially in immunocompromised patients, and associated with infection by opportunistic microorganisms with antimicrobial resistance, such as Klebsiella pneumoniae, highlight the need for the development of new antimicrobial approaches. Thus, the aim of this study was to evaluate the antibacterial and anti-adherent activity of Origanum vulgare L. (oregano) essential oil against Klebsiella pneumoniae strains, as well as the effect of its association with synthetic antimicrobials. To this end, the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) analyses were performed on microdilution plates. The assay of the Minimum Inhibitory Adherence Concentration (MIAC), with test tubes. As well as, the association study through the infusion disc method containing ampicillin (AMP), gentamicin (GEN), ciprofloxacin (CIP) and ceftriaxone (CEF). Therefore, it was possible to obtain that the essential oil of oregano presents antimicrobial and bactericidal activity, with MIC ranging between 128µg/mL and 256 µg/mL and MBC between 256 µg/mL and 512 µg/mL, on the tested K. pneumoniae strains. When used in association with ampicillin and gentamicin, oregano essential oil showed synergistic effect for some strains. Therefore, it is observed that the tested essential oil can act as a promising antibacterial in the treatment of diseases caused by K. pneumoniae.

As bactérias podem ser a causa inicial de determinadas patologias como também um agente secundário responsável pelo desenvolvimento de complicações como as infecções das lesões por pressão. As lesões por pressão configuram um problema de saúde persistente, principalmente em pacientes imunocomprometidos, e associadas a infecção por microrganismos oportunistas com resistência antimicrobiana, como por exemplo a Klebisiella pneumoniae, ressaltam a necessidade do desenvolvimento de novas abordagens antimicrobianas. Assim, o objetivo deste estudo foi avaliar a atividade antibacteriana e antiaderente do óleo essencial de Origanum vulgare L. (orégano) contra cepas da Klebisiella pneumoniae, bem como o efeito da sua associação com antimicrobianos sintéticos. Para isso foram realizadas as análises da Concentração Mínima Inibitória (CIM) e Concentração Bactericida Mínima (CBM) em placas de micro diluição. O ensaio da Concentração Mínima de Aderência (CIMA), com tubos de ensaio. Como também, o estudo de associação através do método de disco de infusão contendo ampicilina, gentamicina, ciprofloxacino e ceftriaxona. Assim sendo, foi possível obter que o óleo essencial de orégano apresenta atividade antimicrobiana e bactericida, com CIM variando entre 128µg/mL e 256 µg/mL e CBM entre 256 µg/mL e 512 µg/mL, sobre as cepas de K. pneumoniae testadas. Quando utilizado em associação com a ampicilina e gentamicina, o óleo essencial de orégano demonstrou efeito sinérgico para algumas cepas. Portanto, observa-se que o óleo essencial testado pode atuar como um promissor antibacteriano no tratamento de doenças causadas por K. pneumoniae.

Evaluation of the antibacterial activity of trans-anethole against Enterococcus cloacae and Enterococcus faecalis strains of food origin / Avaliação da atividade antibacteriana do trans-anetol contra cepas de Enterobacter cloacae e Enterococcus faecalis de origem alimentar

The present study sought to evaluate the antibacterial activity of trans-anethole against food-borne strains of Enterobacter cloacae and Enterococcus faecalis. The study was performed using Minimum Inhibitory Concentration (MIC), and Minimum Bactericidal Concentration (MBC) methods, in addition, disc diffusion technique was used to evaluate the association of trans-anethole with synthetic antimicrobials. Minimum Inhibitory Concentration for Adherence (MICA) testing was also performed. The results revealed that trans-anethole presents no antibacterial activity at any of the concentrations used against the E. cloacae strains tested. However, trans-anethole presented antibacterial effect against five of the six E. faecalis bacterial strains tested, with MIC values ranging from 500 µg/mL to 1000 µg/mL. Further, when analyzing the MBC results against E. faecalis, it was observed that the compound presented values ranging from 500 µg/mL to 1000 µg/mL. As for the associations, it was observed that transanethole when combined with the antimicrobials ampicillin, gentamicin, ciprofloxacin, and ceftriaxone presented synergistic effect against most strains of E. faecalis. However, both trans-anethole and the control chlorhexidine (0.12%) presented no antibiofilm effects against strains of E. faecalis. In short, trans-anethole presented potential antibacterial against E. faecalis strains of food origin, and may upon further study, it may be used alone or in association with synthetic antimicrobials to combat infections caused by this bacterium.

O presente estudo procurou avaliar a atividade antibacteriana do trans-anetol contra cepas de Enterobacter cloacae e Enterococcus faecalis de origem alimentar. O estudo foi realizado utilizando métodos de Concentração Inibitória Mínima (CIM), e Concentração Bactericida Mínima (CBM), além disso, foi utilizada a técnica de difusão de disco para avaliar a associação do trans-anetol com antimicrobianos. O teste de Concentração Inibitória Mínima de Aderência (CIMA) também foi realizado. Os resultados revelaram que o trans-anetol não apresentou atividade antibacteriana em nenhuma das concentrações utilizadas contra as cepas de E. cloacae testadas. No entanto, o trans-anetol apresentou efeito antibacteriano contra cinco das seis cepas bacterianas de E. faecalis testadas, com valores de CIM variando de 500 µg/mL a 1000 µg/mL. Além disso, ao analisar os resultados da CBM contra E. faecalis, observa-se que o composto apresentou valores variando de 500 µg/mL a 1000 µg/mL. Quanto às associações, observou-se que o trans-anetol quando combinado com os antimicrobianos ampicilina, gentamicina, ciprofloxacino, e ceftriaxona apresentou efeito sinérgico contra a maioria das cepas de E. faecalis. No entanto, tanto o trans-anetol quanto o controle clorexidina (0,12%) não apresentaram efeito antibiofilme contra a cepa de E. faecalis. Em suma, o transanetol apresentou potencial antibacteriano contra cepas de E. faecalis de origem alimentar, podendo, mediante estudos mais aprofundados, ser utilizado isoladamente ou em associação com antimicrobianos sintéticos para combater infecções causadas por esta bactéria.

Antioxidant and antimicrobial activities of Thymus vulgaris essential oil contained and synthesis thymus (Vulgaris) silver nanoparticles / Atividades antioxidante e antimicrobiana do óleo essencial de Thymus vulgaris contido e síntese de nanopartículas de prata de timo (Vulgaris)

Several species of thymus have therapeutic properties, so they are used in traditional medicine. In this work was carried out to synthesize Thymus vulgalis silver nanoparticles (TSNPS) and evaluate antioxidant and antimicrobial activities of TSNPS and T. vulgalis essential oil extract (TEOE). The essential oils analyzed by GC-MS and were characterized. Major compounds of phenol, 2 methyl 5 (1 methylethyle) (CAS), thymol and 1,2 Benzene dicarboxylic acid, 3 nitro (CAS) (48.75%, 32.42% and 8.12%, respectively) were detected. Results demonstrated that the TSNPS gave a highest DPPH radical scavenging activity, it was obtained 97.2 at 1000 ug/ml. TSNPS, Thymus + Hexane (T+H), Thymus + Ethanol (T+E) gave the greatest antimicrobial activity than amoxicillin (AM) and ciprofloxacin (CIP). In conclusion: The essential oil of thymus (Vulgaris) and thymus (Vulgaris) silver nanoparticles can be a good source of natural preservatives as an antioxidant and antimicrobial agents for increasing the shelf life of foodstuffs.(AU)

Diversas espécies de timo possuem propriedades terapêuticas, por isso são utilizadas na medicina tradicional. Neste trabalho foi realizado para sintetizar nanopartículas de prata Thymus vulgalis (TSNPS) e avaliar as atividades antioxidante e antimicrobiana de TSNPS e extrato de óleo essencial de T. vulgalis (TEOE). Os óleos essenciais analisados por GC-MS e foram caracterizados. Os principais compostos de fenol, 2 metil 5 (1 metiletilo) (CAS), timol e ácido 1,2 Benzenodicarboxílico, 3 nitro (CAS) (48,75%, 32,42% e 8,12%, respectivamente) foram detectados. Os resultados demonstraram que o TSNPS deu uma maior atividade de eliminação do radical DPPH , foi obtido 97,2 a 1000 ug / ml. TSNPS, Timo + Hexano (T + H), Timo + Etanol (T + E) deu a maior atividade antimicrobiana do que amoxicilina (AM) e ciprofloxacina (CIP). Em conclusão: O óleo essencial de nanopartículas de prata do timo (Vulgaris) e do timo (Vulgaris) pode ser uma boa fonte de conservantes naturais como agentes antioxidantes e antimicrobianos para aumentar a vida útil de alimentos.(AU)

Spectral analysis, in vitro cytotoxicity and antibacterial studies of bioactive principles from the leaves of Conocarpus lancifolius, a common tree of Jazan, Saudi Arabia / Análise espectral, citotoxicidade in vitro e estudos antibacterianos de princípios bioativos das folhas de Conocarpus lancifolius, uma árvore comum de Jazan, Arábia Saudita

The objective of the present study was to analyse the bioactive compounds of the leaves of Conocarpus lancifolius (C. lancifolius). The GC-MS analysis of the hot methanolic extract of the leaves (HMEL) of C. lancifolius exhibited the bioactive compounds such as 1-(3-Methoxy-2-nitrobenzyl) iso quinoline, morphin-4-ol-6,7-dione, 1-bromo N-methyl-, phytol, hexadecanoic acid, 2,3-dihydroxypropyl ester, 2,2:4,2”-terthiophene, ethyl iso-allocholate, caryophyllene oxide, campesterol, epiglobulol, cholestan-3-ol, 2-methylene-, (3á,5à)-, dasycarpidan-1-methanol, acetate (ester) and oleic acid, eicosyl ester. The FT-IR analysis of HMEL of C. lancifolius showed a unique peak at 3184, 2413, 1657 cm-¹ representing coumaric acid, chlorogenic acid and ferulic acid. The HMEL of C. lancifolius was actively inhibiting the proliferation of breast cancer cells MCF-7 ATCC at the concentration of 72.66 ± 8.21 µg/ml as IC50 value. The HMEL of C. lancifolius also revealed a good spectrum of activity against Gram-positive and Gram negative bacterial cultures screened in this work. The activity observed has shown more or less similar effects against screened bacteria. However, the magnitude of potentiality was significantly lesser compared to standard ciprofloxacin disc at p< 0.001 level (99% confidence intervals). Furthermore, the study demonstrating the bioactive compounds can be isolated from the leaves of C. lancifolius.(AU)

O objetivo do presente estudo foi analisar os compostos bioativos das folhas de Conocarpus lancifolius (C. lancifolius). A análise por GC-MS do extrato metanólico quente das folhas (HMEL) de C. lancifolius exibiu os compostos bioativos como 1- (3-Metoxi-2-nitrobenzil) isoquinolina, morfina-4-ol-6,7- diona, 1-bromo-N-metil-, fitol, ácido hexadecanoico, 2,3-di-hidroxipropil éster, 2,2 ‘: 4, 2 ” - tertiofeno, isoalocolato de etil, óxido de cariofileno, campesterol, epiglobulol, colestano -3-ol, 2-metileno-, (3á, 5à) -, dasycarpidan-1-metanol, acetato (éster) e ácido oleico, éster eicosílico. A análise FT-IR de HMEL de C. lancifolius mostrou um pico único em 3184, 2413, 1657 cm-¹ representando ácido cumarico, ácido clorogênico e ácido ferúlico. O HMEL de C. lancifolius inibiu ativamente a proliferação de células de câncer de mama MCF-7 ATCC na concentração de 72,66 ± 8,21 µg/ml como valor de IC50. O HMEL de C. lancifolius também revelou bom espectro de atividade contra culturas de bactérias Gram-positivas e Gram-negativas rastreadas neste trabalho. A atividade observada mostrou efeitos mais ou menos semelhantes contra bactérias rastreadas. No entanto, a magnitude da potencialidade foi significativamente menor em comparação com o disco de ciprofloxacina padrão em nível de p < 0,001 (intervalos de confiança de 99%). Além disso, o estudo demonstrando os compostos bioativos pode ser isolado das folhas de C. lancifolius.(AU)

Chronic superficial infection in a dog caused by multidrug-resistant Pseudomonas aeruginosa

Background: Pseudomonas aeruginosa is a gram-negative aerobic bacterium and non-glucose fermenting, that usually causes opportunistic infections in animals, including humans. It is rarely involved in primary disease. The antibioticresistant bacterial strains are mainly developed due to the inappropriate use of antibiotics, however treating P. aeruginosa infections can be difficult owing to their natural resistance to antibiotics. Furthermorer resistant microorganisms such as P. aeruginosa grow by developing biofilms. Inaccurate diagnoses and absence of adequate microbiological tests can cause difficulties in resolving cases. This report describes a case of chronic superficial infection in a bitch caused by multidrugresistant Pseudomonas aeruginosa (MDR-PA). Case: A 6-year-old bitch Shih Tzu, initially presented with an exudative erythematous lesion in the snout region, which progressed to deep lesions, and spread to the back and limbs; furthermore, the animal always experienced a fever before new wounds emerged. Lesion samples, collected using a swab and processed at the Veterinary Microbiology Laboratory of the Federal University of Jatai (UFJ), revealed the presence of Pseudomonas aeruginosa. The isolate was multidrug-resistant and a carrier of TEM and ppyR genes. In the diffusion disk antibiogram, the isolate was found resistant to 14 different antibiotics belonging to 6 classes. Antimicrobial resistance was also tested using the minimum inhibitory concentration (MIC) test against imipenem, ceftazidime, ciprofloxacin, ticarcillin + clavulanic acid and aztreonam present in the MIC test strip. Treatment with amikacin and muporicin proved to be effective; however, owing to lesions extending to the face and palpebral involvement, the animal lost its eyeballs. Discussion: Pseudomonas aeruginosa is frequently associated with nosocomial infections mainly affecting immunosuppressed patients. Among the antibiotics tested, the group with the highest number of ineffective antibiotics was beta-lactams, where sensitivity was only observed for ticarcillin and ceftazidime. Recent studies have demonstrated that ceftazidime can reduce biofilm volume, inhibit motility, and repress the expression of genes associated with bacterial adhesion in P. aeruginosa. Therefore, the production of biofilm in P. aeruginosa is an important virulence factor as it facilitates a stable environment for the microorganism, which protects the bacteria from contact with antimicrobials. In addition, prolonged exposure to a wide variety of antimicrobials creates an environment of selective pressure between microorganisms, facilitating the emergence of multidrug-resistant strains. Furthermore, it is now well recognized that low doses of antibiotics, administered during continuous and fluctuating treatments, can stimulate biofilm establishment and are partly responsible for biofilm-specific antimicrobial tolerance. The resistance profile of P. aeruginosa isolated from dogs varies considerably, and the presence of isolates with a possible biofilm production capacity represents a challenge for the interpretation of the antimicrobial susceptibility profile. Culture and antibiogram is fundamentally important, both clinically and in environmental monitoring, in addition to the use of antibiogram data for decision making in clinical treatment.

Corneal ulcer in a sheep - treatment with third eyelid flap

Background: Corneal ulcers are frequently reported in the literature in several species, however, the treatment of traumaticlesions on the corneal surface of ruminants is still poorly described. The use of the third eyelid flap is questioned whenapplied to deep ulcers, since the technique prevents the evolutionary follow-up of corneal healing and compromises care.However, several authors report its successful use for the treatment of superficial and deep corneal ulcers. This techniqueprotects the ocular surface and prevents the occurrence of new lesions or their aggravation. The aim of this study was toreport the treatment of corneal ulcers in sheep through the third eyelid flap associated with the use of topical antibiotics.Case: A 2-year-old male Dorper sheep, with ocular discomfort, conjunctival hyperemia and mucopurulent secretion in theleft eye, was assisted in a private rural property in the West region of Bahia, Brazil. The owner reported that before seeking veterinary help he used a spray based on oxytetracycline and hydrocortisone, which worsened the animal’s condition.Upon inspection, it was observed eyelid asymmetry, slight lateralization of the head to the left, periorbital alopecia onthe left face, intense blepharospasm in the left eye, with projection of the third eyelid occurring at times. The ophthalmicexamination was performed after application of anesthetic eye drops based on proxymetacaine to reduce eye discomfort.A corneal stromal lesion, edema and fibrovascular tissue in the left eye were identified, but the cause of the lesion was notdefined, with lagophthalmia, entropion, dystychiasis or ectopic cilia being ruled out. So, it was suggested that the lesionhad occurred by a foreign body, such as dust...

Corneal ulcer in a sheep - treatment with third eyelid flap

Background: Corneal ulcers are frequently reported in the literature in several species, however, the treatment of traumaticlesions on the corneal surface of ruminants is still poorly described. The use of the third eyelid flap is questioned whenapplied to deep ulcers, since the technique prevents the evolutionary follow-up of corneal healing and compromises care.However, several authors report its successful use for the treatment of superficial and deep corneal ulcers. This techniqueprotects the ocular surface and prevents the occurrence of new lesions or their aggravation. The aim of this study was toreport the treatment of corneal ulcers in sheep through the third eyelid flap associated with the use of topical antibiotics.Case: A 2-year-old male Dorper sheep, with ocular discomfort, conjunctival hyperemia and mucopurulent secretion in theleft eye, was assisted in a private rural property in the West region of Bahia, Brazil. The owner reported that before seeking veterinary help he used a spray based on oxytetracycline and hydrocortisone, which worsened the animals condition.Upon inspection, it was observed eyelid asymmetry, slight lateralization of the head to the left, periorbital alopecia onthe left face, intense blepharospasm in the left eye, with projection of the third eyelid occurring at times. The ophthalmicexamination was performed after application of anesthetic eye drops based on proxymetacaine to reduce eye discomfort.A corneal stromal lesion, edema and fibrovascular tissue in the left eye were identified, but the cause of the lesion was notdefined, with lagophthalmia, entropion, dystychiasis or ectopic cilia being ruled out. So, it was suggested that the lesionhad occurred by a foreign body, such as dust...(AU)

Microbial source tracking and antimicrobial resistance in one river system of a rural community in Bahia, Brazil

Abstract Use of antibiotics inevitably leads to antimicrobial resistance. Selection for resistance occurs primarily within the gut of humans and animals as well as in the environment through natural resistance and residual antibiotics in streams and soil. We evaluated antimicrobial resistance in Gram negative bacteria from a river system in a rural community in Bahia, Brazil. Water was collected from the Jiquiriçá and Brejões rivers and the piped water supply. Additionally, stools were collected from a random sample of residents, cows, pigs and horses near the river. The samples were screened for bacteria resistant to ciprofloxacin, cefotaxime, and meropenem and identified biochemically at the genus and species levels. Microbial source tracking demonstrated that ruminant and human fecal contamination increased as the rivers neared the village center and decreased after the last residence. Antibiotic bacteria were identified from all samples (n = 32). No bacteria were resistant to carbapenems, but the majority of the enterobacteria were resistant to ciprofloxacin, even though this class of antibiotics is not commonly used in food animals in this region. Considering these facts, together with the pattern of human fecal contamination, a human source was considered most likely for these resistant isolates.

Resumo O uso de antibióticos inevitavelmente leva à resistência antimicrobiana. A seleção para resistência antimicrobiana ocorre principalmente no intestino de seres humanos e animais, bem como no meio ambiente, através da resistência natural e resíduos de antibióticos nos esgotos e no solo. Avaliamos a resistência antimicrobiana em bactérias Gram-negativas de um sistema fluvial em uma comunidade rural da Bahia, Brasil. A água foi coletada nos rios Jiquiriçá e Brejões e no abastecimento de água encanada. Além disso, foram coletadas amostras randomizadas de fezes de moradores, vacas, porcos e cavalos próximos ao rio. As amostras foram triadas para bactérias resistentes à ciprofloxacina, cefotaxima e meropenem e identificadas bioquimicamente nos níveis de gênero e espécie. O rastreamento de fontes microbianas demonstrou que a contaminação fecal de ruminantes e humanos aumentou à medida que os rios se aproximavam do centro da vila e diminuía após a última residência. Bactérias resistentes a antibióticos foram identificadas em todas as amostras (n = 32). Nenhuma bactéria demonstrou ser resistente aos carbapenêmicos testados, contudo, foi encontrado enterobactérias resistentes à ciprofloxacina, ainda que essa classe de antibióticos não seja comumente usada na medicina veterinária dos animais dessa região. Considerando esses fatos, juntamente com o padrão de contaminação fecal avaliado, a fonte de contaminação humana foi considerada a mais provável na interação desses isolados resistentes.

Microbial source tracking and antimicrobial resistance in one river system of a rural community in Bahia, Brazil / Rastreamento de fontes microbianas e resistência antimicrobiana em um sistema fluvial de uma comunidade rural da Bahia, Brasil

Use of antibiotics inevitably leads to antimicrobial resistance. Selection for resistance occurs primarily within the gut of humans and animals as well as in the environment through natural resistance and residual antibiotics in streams and soil. We evaluated antimicrobial resistance in Gram negative bacteria from a river system in a rural community in Bahia, Brazil. Water was collected from the Jiquiriçá and Brejões rivers and the piped water supply. Additionally, stools were collected from a random sample of residents, cows, pigs and horses near the river. The samples were screened for bacteria resistant to ciprofloxacin, cefotaxime, and meropenem and identified biochemically at the genus and species levels. Microbial source tracking demonstrated that ruminant and human fecal contamination increased as the rivers neared the village center and decreased after the last residence. Antibiotic bacteria were identified from all samples (n = 32). No bacteria were resistant to carbapenems, but the majority of the enterobacteria were resistant to ciprofloxacin, even though this class of antibiotics is not commonly used in food animals in this region. Considering these facts, together with the pattern of human fecal contamination, a human source was considered most likely for these resistant isolates.

O uso de antibióticos inevitavelmente leva à resistência antimicrobiana. A seleção para resistência antimicrobiana ocorre principalmente no intestino de seres humanos e animais, bem como no meio ambiente, através da resistência natural e resíduos de antibióticos nos esgotos e no solo. Avaliamos a resistência antimicrobiana em bactérias Gram-negativas de um sistema fluvial em uma comunidade rural da Bahia, Brasil. A água foi coletada nos rios Jiquiriçá e Brejões e no abastecimento de água encanada. Além disso, foram coletadas amostras randomizadas de fezes de moradores, vacas, porcos e cavalos próximos ao rio. As amostras foram triadas para bactérias resistentes à ciprofloxacina, cefotaxima e meropenem e identificadas bioquimicamente nos níveis de gênero e espécie. O rastreamento de fontes microbianas demonstrou que a contaminação fecal de ruminantes e humanos aumentou à medida que os rios se aproximavam do centro da vila e diminuía após a última residência. Bactérias resistentes a antibióticos foram identificadas em todas as amostras (n = 32). Nenhuma bactéria demonstrou ser resistente aos carbapenêmicos testados, contudo, foi encontrado enterobactérias resistentes à ciprofloxacina, ainda que essa classe de antibióticos não seja comumente usada na medicina veterinária dos animais dessa região. Considerando esses fatos, juntamente com o padrão de contaminação fecal avaliado, a fonte de contaminação humana foi considerada a mais provável na interação desses isolados resistentes.

Microbial source tracking and antimicrobial resistance in one river system of a rural community in Bahia, Brazil / Rastreamento de fontes microbianas e resistência antimicrobiana em um sistema fluvial de uma comunidade rural da Bahia, Brasil

Use of antibiotics inevitably leads to antimicrobial resistance. Selection for resistance occurs primarily within the gut of humans and animals as well as in the environment through natural resistance and residual antibiotics in streams and soil. We evaluated antimicrobial resistance in Gram negative bacteria from a river system in a rural community in Bahia, Brazil. Water was collected from the Jiquiriçá and Brejões rivers and the piped water supply. Additionally, stools were collected from a random sample of residents, cows, pigs and horses near the river. The samples were screened for bacteria resistant to ciprofloxacin, cefotaxime, and meropenem and identified biochemically at the genus and species levels. Microbial source tracking demonstrated that ruminant and human fecal contamination increased as the rivers neared the village center and decreased after the last residence. Antibiotic bacteria were identified from all samples (n = 32). No bacteria were resistant to carbapenems, but the majority of the enterobacteria were resistant to ciprofloxacin, even though this class of antibiotics is not commonly used in food animals in this region. Considering these facts, together with the pattern of human fecal contamination, a human source was considered most likely for these resistant isolates.(AU)

O uso de antibióticos inevitavelmente leva à resistência antimicrobiana. A seleção para resistência antimicrobiana ocorre principalmente no intestino de seres humanos e animais, bem como no meio ambiente, através da resistência natural e resíduos de antibióticos nos esgotos e no solo. Avaliamos a resistência antimicrobiana em bactérias Gram-negativas de um sistema fluvial em uma comunidade rural da Bahia, Brasil. A água foi coletada nos rios Jiquiriçá e Brejões e no abastecimento de água encanada. Além disso, foram coletadas amostras randomizadas de fezes de moradores, vacas, porcos e cavalos próximos ao rio. As amostras foram triadas para bactérias resistentes à ciprofloxacina, cefotaxima e meropenem e identificadas bioquimicamente nos níveis de gênero e espécie. O rastreamento de fontes microbianas demonstrou que a contaminação fecal de ruminantes e humanos aumentou à medida que os rios se aproximavam do centro da vila e diminuía após a última residência. Bactérias resistentes a antibióticos foram identificadas em todas as amostras (n = 32). Nenhuma bactéria demonstrou ser resistente aos carbapenêmicos testados, contudo, foi encontrado enterobactérias resistentes à ciprofloxacina, ainda que essa classe de antibióticos não seja comumente usada na medicina veterinária dos animais dessa região. Considerando esses fatos, juntamente com o padrão de contaminação fecal avaliado, a fonte de contaminação humana foi considerada a mais provável na interação desses isolados resistentes.(AU)

Canine cystitis - biofilm formation by bacterial isolates

Background: Biofilms have been reported as important virulent markers associated with drug resistance in urinary tractinfections (UTIs) in humans and dogs. However, in veterinary medicine, researches involving biofilm formation, treatments and preventions have been limited; yet, it is still possible to find few studies demonstrating biofilm-forming bacteriaassociated with different comorbidities such as otitis, wound infections, UTIs, and endometritis. These studies generallyselect dogs with chronic and recurrent infections, which could be an important factor in antibiotic resistance. We aimed toevaluate biofilms in sporadic cystitis regarding prevalence and drug resistance.Materials, Methods & Results: Urine samples were collected by cystocentesis from 36 client-owned dogs under clinicaland laboratory suspicion of non-recurrent urinary bladder infection (cystitis). Urine was aseptically plated onto bloodagar, MacConkey, and CLED, followed by incubation for 24 to 48 h. Definitive identification of a potential pathogen wasmade by subculture collected from an isolated colony to obtain a pure culture. The gram staining method and specificbiochemical tests (phenol red fermentation, lysine, phenylalanine, citrate, sulfide-indole-motility, and urease) were usedto distinguish and classify the bacteria. After identification, the bacteria were tested for antimicrobial susceptibility by astandard disk diffusion method, using the following antimicrobials: amoxicillin with clavulanic acid, ampicillin, ceftriaxone, ciprofloxacin, clindamycin, cefazolin, cephalothin, erythromycin, gentamicin, norfloxacin, and sulfamethoxazoletrimethoprim. The biofilm-forming ability was determined based on a culture...

Phenotypic and molecular characterization of fluoroquinolone resistant Pseudomonas aeruginosa isolates in Palestine

Abstract Fluoroquinolones are important antimicrobial agents for the treatment of Pseudomonas infections. A total of 11 isolates of P. aeruginosa were collected from different clinical samples from different medical centers in the North West Bank-Palestine during 2017. In this study, resistance to fluoroquinolones and secretions of -lactamases were detected by phenotypic methods, while presence of -lactamase gene sequences and other virulence factors were detected by PCR technique. PCR product for gyrA, parC and parE genes were sequenced for further analyses. The phylogenetic analyses, population diversity indices and haplotypes determination were conducted using computer programs MEGA version 6, DnaSP 5.1001 and median-joining algorithm in the program Network 5, respectively. Results of this study showed that the MIC for ciprofloxacin and norfloxacin had a range of 32-256 µg/ml. In addition, all isolates carried either exoT or exoT and exoY genes, different -lactamase genes and 82% of these isolates harbored class 1 integrons. Analyses of the gyrA, parC and parE sequences were found to be polymorphic, had high haplotype diversity (0.945-0.982), low nucleotide diversity (0.01225-0.02001) and number of haplotypes were 9 for each gyrA and parE genes and 10 haplotypes for parC gene. The founder haplotypes being Hap-1 (18%), Hap-2 (27.3%) and Hap-6 (9.1%) for gyrA, parC and parE genes, respectively. Two of ParE haplotypes were detected as indel haplotypes. The Median-joining- (MJ) networks constructed from haplotypes of these genes showed a star-like expansion. The neutrality tests (Tajimas D test and Fus Fs test) for these genes showed negative values. Palestinian fluoroquinolone resistant P. aeruginosa strains showed high MIC level for fluoroquinolones, -lactamase producers, carried type III secretion exotoxin-encoding genes, most of them had integrase I gene and had high level of mutations in QRDR regions in gyrA, parC and parE genes. All these factors may play an important role in the invasiveness of these strains and make them difficult to treat. Isolation of these strains from different medical centers, indicate the need for a strict application of infection control measures in Medical centers in the North West Bank-Palestine that aim to reduce expense and damage caused by P. aeruginosa infections. Molecular analyses showed that Palestinian fluoroquinolone resistant P. aeruginosa haplotypes are not genetically differentiated; however, more mutations may exist in these strains.

Resumo Fluoroquinolonas são agentes antimicrobianos importantes para o tratamento de infecções por Pseudomonas. Um total de 11 bacilos isolados de P. aeruginosa foram coletados de diferentes amostras clínicas provenientes de diferentes centros médicos na Cisjordânia-Palestina durante o ano de 2017. Neste estudo, resistência a fluoroquinolonas e secreções de -lactamases foram detectadas por métodos fenotípicos, enquanto a presença de sequências do gene -lactamase e outros fatores de virulência foram detectados pela técnica de PCR (Proteína C-reativa). O produto de PCR para os genes gyrA, parC e parE foram sequenciados para análises posteriores. As análises filogenéticas, os índices de diversidade populacional e a determinação de haplótipos foram realizados utilizando os softwares MEGA versão 6, DnaSP 5.1001 e o algoritmo de junção de mediana do programa Network 5, respectivamente. Os resultados deste estudo mostraram que a MIC para ciprofloxacina e norfloxacina tinha um intervalo de 32-256 µg/ml. Além disso, todos os bacilos isolados carregavam genes exoT ou exoT e exoY, genes de -lactamase diferentes e 82% desses isolados continham integrons de classe 1. As análises das sequências gyrA, parC e parE foram consideradas polimórficas, com alta diversidade de haplótipos (0,945-0,982), baixa diversidade de nucleotídeos (0,01225-0,02001) e o número de haplótipos foi de 9 para cada gene de gyrA e parE e 10 haplótipos para o gene parC. Os haplótipos fundadores são Hap-1 (18%), Hap-2 (27,3%) e Hap-6 (9,1%) para os genes gyrA, parC e parE, respectivamente. Dois dos haplótipos parE foram detectados como haplótipos InDel. As redes Median-joining (MJ) construídas a partir de haplótipos desses genes mostraram uma expansão semelhante à de uma estrela. Os testes de neutralidade (teste D de Tajima e teste Fs de Fu) para esses genes apresentaram valores negativos. As cepas palestinas de P. aeruginosa resistentes a fluoroquinolonas mostraram alto nível de MIC para fluoroquinolonas, produtores de -lactamase, genes codificadores de exotoxina de secreção tipo III, a maioria deles tinha o gene integrase I e tinha alto nível de mutações nas regiões QRDR nos genes gyrA, parC e parE. Todos esses fatores podem desempenhar um papel importante na invasão dessas cepas e torná-las difíceis de tratar. O isolamento dessas cepas em diferentes centros médicos, indica a necessidade de uma aplicação estrita de medidas de controle de infecção em centros médicos da Cisjordânia-Palestina que visam reduzir despesas e danos causados por infecções por P. aeruginosa. As análises moleculares mostraram que os haplótipos de P. aeruginosa resistentes à fluoroquinolona palestina não são geneticamente diferenciados; no entanto, mais mutações podem existir nessas cepas.