RESUMEN

Complex oviposition decisions allow gravid Aedes aegypti mosquitoes to select suitable sites for egg-laying to increase the probability that their progeny will thrive. The bacterial communities present in larval niches influence mosquito oviposition behavior, and gravid mosquitoes transmit key microbial associates to breeding sites during oviposition. Our study evaluated whether symbiotic Klebsiella sp., which are strongly associated with mosquitoes, emit volatiles that affect mosquito oviposition decisions. Dual-choice behavioral assays demonstrated that volatile organic compounds emitted by Klebsiella sp. induce a preference in oviposition decisions by Ae. aegypti. Bacterial headspace volatiles were sampled by solid-phase microextraction, and subsequent combined gas chromatography and electroantennogram detection analysis, revealed that the antennae of gravid females detect two compounds present in the Klebsiella sp. headspace. These compounds were identified by gas chromatography and mass spectrometry as 2-ethyl hexanol and 2,4-di­tert-butylphenol. The binary blend of these compounds elicited a dose-dependent egg-laying preference by gravid mosquitoes. We propose that bacterial symbionts, which are associated with gravid mosquitoes and may be transferred to aquatic habitats during egg-laying, together with their volatiles act as oviposition cues indicating the suitability of active breeding sites to conspecific females.

Asunto(s)

Aedes , Femenino , Animales , Aedes/fisiología , Odorantes , Cromatografía de Gases y Espectrometría de Masas , Oviposición , Bacterias

RESUMEN

Klebsiella sp. strain AqSCr, isolated from Cr(VI)-polluted groundwater, reduces Cr(VI) both aerobically and anaerobically and resists up 34 mM Cr(VI); this resistance is independent of the ChrA efflux transporter. In this study, we report the whole genome sequence and the transcriptional profile by RNA-Seq of strain AqSCr under Cr(VI)-adapted conditions and found 255 upregulated and 240 downregulated genes compared to controls without Cr(VI) supplementation. Genes differentially transcribed were mostly associated with oxidative stress response, DNA repair and replication, sulfur starvation response, envelope-osmotic stress response, fatty acid (FA) metabolism, ribosomal subunits, and energy metabolism. Among them, genes not previously associated with chromium resistance, for example, cybB, encoding a putative superoxide oxidase (SOO), gltA2, encoding an alternative citrate synthase, and des, encoding a FA desaturase, were upregulated. The sodA gene encoding a manganese superoxide dismutase was upregulated in the presence of Cr(VI), whereas sodB encoding an iron superoxide dismutase was downregulated. Cr(VI) resistance mechanisms in strain AqSCr seem to be orchestrated by the alternative sigma factors fecl, rpoE, and rpoS (all of them upregulated). Membrane lipid analysis of the Cr(IV)-adapted strain showed a lower proportion of unsaturated lipids with respect to the control, which we hypothesized could result from unsaturated lipid peroxidation followed by degradation, together with de novo synthesis mediated by the upregulated FA desaturase-encoding gene, des. This report helps to elucidate both Cr(VI) toxicity targets and global bacterial response to Cr(VI).

RESUMEN

Objetivo. Determinar las características citológicas y microbiológicas de lavados uterinos de bajo volúmen en hembras con problemas reproductivos en dos hatos del Altiplano Cundiboyacense. Materiales y métodos. Se evaluaron 21 muestras de lavados uterinos de vacas con historia de problemas reproductivos procedentes de dos hatos lecheros, en el primer hato ubicado en Sibaté (Cundinamarca) se muestrearon 10 animales y en el segundo ubicado en Ventaquemada (Boyacá) se obtuvieron las 11 muestras restantes. Se realizó evaluación macroscópica y microscópica de cada uno de los lavados uterinos; para la evaluación citológica se utilizaron las tinciones de Gram y Wright. La determinación de la flora bacteriana se realizó mediante técnicas estándar en microbiología para la identificación de bacterias no exigentes, adicionalmente se realizaron pruebas de sensibilidad por la técnica de difusión de Kirby-bauer. Resultados. En la citología se observaron bacilos y cocos gram positivos, células epiteliales de descamación uterina (CEDU) y respuesta inflamatoria (RI) mediada por polimorfonucleares (PMN). Los aislamientos bacterianos correspondieron a Lactobacillus sp como flora bacteriana normal del tracto genital (16.66%) y Klebsiella sp como flora acompañante (16.66%); adicionalmente se aislaron bacterias patógenas causantes de problemas reproductivos como Streptococcus sp. β hemolítico (33.33%), Streptococcus sp α hemolítico (50%) y Streptococcus sp. γ hemolítico (50%) y Arcanobacterium pyogenes (Corynebacterium-Actinomyces pyogenes) (16.66%); en un 9.52% de las muestras se observaron espiroquetas. Conclusiones. Se observó una relación directa entre el aspecto macroscópico de los lavados uterinos, la respuesta citológica y el aislamiento de bacterias.

Asunto(s)

Bovinos , Animales , Arcanobacterium , Bovinos , Klebsiella , Streptococcus , Enfermedades Uterinas

RESUMEN

Objective: To evaluate the association between Escherischia Coli (E. Coli) and Klebsiella spp bacteremia, both of which produce Extended Spectrum beta-lactamases (ESBL), and the use of third generation cephalos-porines (TGC) during 30 days prior to the development of the infection. Patients and Methods: Case study of all patients hospitalized between 2004 and 2007 at Hospital de Niños Roberto del Río, who presented E. coli y Klebsiella spp bacteremia. Results: 212 episodes of E. coli o Klebsiella spp bacteremias were found; 205 contained all necessary data, 47 of which were cases and 158 were controls. The use of TGC showed a statistically significant correlation with the finding of ESBL in those patients' bacteremias (OR 2,5; IC95 percent 1,26-5). In addition, an independent statistically significant association was found with hospital acquired infections (OR 8,2; IC95 percent 2,7-27,1), and the use of Central Venous Catheter (OR 38,9; IC95 percent 14,3-106,0) became close to statistical significance. Conclusions: This study shows a statistically significant association between ESBL bacteremias and use of TGC within 30 days prior to the infection, confirming other literature reports.

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Objetivo: Estudiar la asociación entre bacteriemia por E. coli y Klebsiella spp productoras de beta-lactamasas de espectro extendido (BLEE) y el uso de cefalosporinas de tercera generación (CTG) durante los 30 días previos al desarrollo de la infección. Método: Estudio caso control encestado desarrollado entre los años 2004 y 2007 en el Hospital de Niños Roberto del Río, con análisis retrospectivo de los pacientes con bacteriemias por E. coli y Klebsiella spp. Resultados: Se registraron 212 episodios de bacteriemias por E. coli o Klebsiella spp, en 205 de ellos se contó con los registros necesarios, 47 fueron casos y 158 controles. El uso de CTG se asoció de modo estadísticamente significativo con la probabilidad de aislar cepas BLEE de bacteriemias en estos pacientes (OR 2,5; IC95 por ciento 1,26-5). Además, se observó asociación independiente y estadísticamente significativa con Infección Intrahospitalaria (OR 8,2; IC95 por ciento 2,7-27,1), y el uso de Catéter Venoso Central (OR 38,9; IC95 por ciento 14,3-106,0) se acercó a la significancia estadística. Conclusiones: Nuestro trabajo establece la asociación estadística entre bacteriemia por cepas BLEE y el uso de CTG dentro de los 30 días previos al desarrollo de la infección, confirmando lo sugerido por la literatura.

Asunto(s)

Humanos , Masculino , Adolescente , Femenino , Recién Nacido , Lactante , Preescolar , Niño , Bacteriemia/microbiología , Cefalosporinas/efectos adversos , Infecciones por Escherichia coli/etiología , Infecciones por Klebsiella/etiología , beta-Lactamasas/metabolismo , Resistencia betalactámica , Bacteriemia/epidemiología , Estudios de Casos y Controles , Infección Hospitalaria , Cateterismo Venoso Central/efectos adversos , Cefalosporinas/uso terapéutico , Escherichia coli/aislamiento & purificación , Infecciones por Escherichia coli/epidemiología , Infecciones por Klebsiella/epidemiología , Klebsiella/aislamiento & purificación , Factores de Riesgo

RESUMEN

Non-symbiotic diazotrophic bacteria are amongst the most important functional groups of soil-dwelling microorganisms. These bacteria contribute to plant growth predominantly through biological N2 fixation. Here, we evaluated the density and diversity of non-symbiotic diazotrophic bacteria in soils taken from diverse land use systems (LUS) in Amazonia using nitrogen-free media. A total of 30 soil samples were collected from the following LUS: pristine forest, young secondary forest, old secondary forest, agroforestry, agriculture and pasture. Bacterial density was evaluated by the most probable number (MPN) method utilizing N-free semi-solid media with varied compositions (JNFb, NFb, LGI and Fam). Individual isolates were characterized by colony and cellular morphology as well as total protein profiles and nitrogenase activity. Isolate genotypes were determined by partial 16S rDNA sequences. No typical diazotrophic growth in the JNFb medium was observed. Bacterial densities in the NFb medium were higher in the agriculture and agroforestry soil samples. In LGI and Fam media, bacterial densities were highest in the pasture soil samples. Overall, 22 isolates with high phenotypic diversity were obtained. Eleven isolates exhibited nitrogenase activity. Sequences of 16S rDNA genes of 14 out of 19 isolates had similarities below 100 % with known nitrogen-fixing species. Isolates were identified as belonging to the Burkholderia, Enterobacter, Serratia, Klebsiella, and Bacillus genera. A higher number of isolates from pasture soil samples were isolated, with the majority of these belonging to the Burkholderia and Bacillus genera. Among the isolates, unknown sequences were obtained, possibly indicating new species. Taken together, these data demonstrate that Fam, NFb, and LGI semi-solid media allowed the growth of diazotrophic bacteria belonging to different phylogenetic lines.

RESUMEN

Non-symbiotic diazotrophic bacteria are amongst the most important functional groups of soil-dwelling microorganisms. These bacteria contribute to plant growth predominantly through biological N2 fixation. Here, we evaluated the density and diversity of non-symbiotic diazotrophic bacteria in soils taken from diverse land use systems (LUS) in Amazonia using nitrogen-free media. A total of 30 soil samples were collected from the following LUS: pristine forest, young secondary forest, old secondary forest, agroforestry, agriculture and pasture. Bacterial density was evaluated by the most probable number (MPN) method utilizing N-free semi-solid media with varied compositions (JNFb, NFb, LGI and Fam). Individual isolates were characterized by colony and cellular morphology as well as total protein profiles and nitrogenase activity. Isolate genotypes were determined by partial 16S rDNA sequences. No typical diazotrophic growth in the JNFb medium was observed. Bacterial densities in the NFb medium were higher in the agriculture and agroforestry soil samples. In LGI and Fam media, bacterial densities were highest in the pasture soil samples. Overall, 22 isolates with high phenotypic diversity were obtained. Eleven isolates exhibited nitrogenase activity. Sequences of 16S rDNA genes of 14 out of 19 isolates had similarities below 100 % with known nitrogen-fixing species. Isolates were identified as belonging to the Burkholderia, Enterobacter, Serratia, Klebsiella, and Bacillus genera. A higher number of isolates from pasture soil samples were isolated, with the majority of these belonging to the Burkholderia and Bacillus genera. Among the isolates, unknown sequences were obtained, possibly indicating new species. Taken together, these data demonstrate that Fam, NFb, and LGI semi-solid media allowed the growth of diazotrophic bacteria belonging to different phylogenetic lines.

RESUMEN

OBJECTIVE: This work aimed to evaluate the effect of fructooligosaccharide and inulin added to an enteral diet on the translocation of Klebsiella sp. in mice. METHODS: Four- to six-week-old Swiss albino mice were divided into nine groups and fed enteral diets containing different combinations of fructooligosaccharide, inulin, antibiotic and corticoid, inoculated or not with Klebsiella pneumoniae. On day 5, the animals of four groups were fed an enteral diet contaminated with approximately 10(10)CFU/g of K. pneumoniae. At defined times, two animals of each group were sacrificed and their organs (spleen, heart, liver, lungs, and kidneys) were aseptically collected, weighed, and analyzed for the presence of typical Klebsiella sp. colonies. RESULTS: A higher number of CFU/g of Klebsiella was detected in the organs of the animals in the immune-suppressed group fed the diet contaminated with K. pneumoniae and without prebiotics. Animals fed the diet enriched with fructooligosaccharide and inulin, at a concentration of 15.3mg/g of body weight, had a shorter period of Klebsiella sp. translocation, compared with those not fed prebiotics in the diet. CONCLUSION: The addition of fructooligosacharide and inulin in enterais diets at a concentration of 15.3mg/g of body weight resulted in the reduction of translocation of Klebsiella for spleen, heart, liver, lung and kidneys of mice that had received the diet contaminated associated or not with antibiotic and imunodepressor drug.

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OBJETIVO: Avaliar o efeito da administração de frutooligossacarídeo e inulina, adicionados à dieta enteral, na translocação de Klebsiella sp. em camundongos. MÉTODOS: Camundongos albinos suíços, com quatro a seis semanas de vida, foram divididos em nove grupos e tratados com dietas enterais contendo diferentes combinações de frutooligossacarídeos, inulina, antibiótico e corticóide, inoculadas ou não com Klebsiella pneumoniae. No quinto dia de experimento, os animais dos tratamentos IV, V, VIII e IX foram alimentados com dieta enteral contaminada com, aproximadamente, 10(10)UFC/g de Klebsiella pneumoniae. Em tempos definidos (sexto, sétimo, nono, décimo primeiro e décimo terceiro dia de experimento), dois animais de cada grupo foram sacrificados e seus órgãos: baço, coração, fígado, pulmões e rins, foram coletados assepticamente, pesados e analisados quanto à presença de colônias típicas de Klebsiella sp. RESULTADOS: Maior número de UFC/g de Klebsiella sp. foi detectado em órgãos de animais do grupo imunodeprimidos, que receberam dieta com Klebsiella pneumoniae e sem prebióticos. Animais que receberam dieta enriquecida com frutooligossacarídeo e inulina, na concentração de 15,3mg/g de peso corporal, apresentaram menor período para a translocação de Klebsiella sp., quando comparados aos animais que não receberam prebióticos na dieta. CONCLUSÃO: A adição de frutooligossacarídeos e inulina em dietas enterais na dose de 15,3mg/g de peso corporal resultou na diminuição de translocação de Klebsiella para baço, coração, fígado, pulmão e rins de camundongos que receberam a dieta contaminada associada ou não com droga imunodepressora e antibiótico.

Asunto(s)

Animales , Ratones , Inulina , Klebsiella pneumoniae , Nutrición Enteral/métodos , Traslocación Bacteriana

RESUMEN

The strain Klebsiella sp. K18 produces the enzyme glucosyltransferase and catalyses the conversion of sucrose to palatinose, an alternative sugar that presents low cariogenicity. Response Surface Methodology was successfully employed to determine the optimal concentration of culture medium components. Maximum glucosyltransferase production (21.78 U mL-1) was achieved using the optimized medium composed by sugar cane molasses (80 g L-1), bacteriological peptone (7 g L-1) and yeast extract (20 g L-1), after 8 hours of fermentation at 28ºC. The conversion of sucrose to palatinose was studied utilizing immobilized cells in calcium alginate. The effects of the alginate concentration (2-4%), cell mass concentration (20-40%) and substrate concentration (25-45%) were evaluated and the yield of palatinose was approximately 62.5%.

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A linhagem Klebsiella sp. K18 produz a enzima glicosiltransferase que catalisa a conversão de sacarose em palatinose, um açúcar alternativo que apresenta baixa cariogenicidade. Metodologia de Superfície de Resposta foi empregada com sucesso para determinar a concentração ótima dos componentes do meio de cultivo. A máxima produção deglicosiltransferase (21,78 U mL-1) foi obtida utilizando o meio de cultivo otimizado composto por melaço de cana de açúcar (80 g L-1), peptona bacteriológica (7 g L-1) e extrato de levedura (20 g L-1), após 8 horas de fermentação a 28ºC. A conversão desacarose em palatinose foi estudada utilizando células imobilizadas em alginato de cálcio. Os efeitos da concentração de alginato (2-4%), concentração de massa celular (20-40%) e concentração de substrato (25-45%) foram avaliados e a porcentagem de palatinose foi de aproximadamente 62,5%.

Asunto(s)

Alginatos , Cariogénicos , Fermentación , Glicosiltransferasas/análisis , Técnicas In Vitro , Klebsiella/enzimología , Melaza/análisis , Sacarosa/análisis , Saccharum/enzimología , Cromatografía Líquida de Alta Presión , Métodos , Métodos

RESUMEN

The strain Klebsiella sp. K18 produces the enzyme glucosyltransferase and catalyses the conversion of sucrose to palatinose, an alternative sugar that presents low cariogenicity. Response Surface Methodology was successfully employed to determine the optimal concentration of culture medium components. Maximum glucosyltransferase production (21.78 U mL(-1)) was achieved using the optimized medium composed by sugar cane molasses (80 g L(-1)), bacteriological peptone (7 g L(-1)) and yeast extract (20 g L(-1)), after 8 hours of fermentation at 28°C. The conversion of sucrose to palatinose was studied utilizing immobilized cells in calcium alginate. The effects of the alginate concentration (2-4%), cell mass concentration (20-40%) and substrate concentration (25-45%) were evaluated and the yield of palatinose was approximately 62.5%.

RESUMEN

The strain Klebsiella sp. K18 produces the enzyme glucosyltransferase and catalyses the conversion of sucrose to palatinose, an alternative sugar that presents low cariogenicity. Response Surface Methodology was successfully employed to determine the optimal concentration of culture medium components. Maximum glucosyltransferase production (21.78 U mL-1) was achieved using the optimized medium composed by sugar cane molasses (80 g L-1), bacteriological peptone (7 g L-1) and yeast extract (20 g L-1), after 8 hours of fermentation at 28°C. The conversion of sucrose to palatinose was studied utilizing immobilized cells in calcium alginate. The effects of the alginate concentration (2-4%), cell mass concentration (20-40%) and substrate concentration (25-45%) were evaluated and the yield of palatinose was approximately 62.5%.

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A linhagem Klebsiella sp. K18 produz a enzima glicosiltransferase que catalisa a conversão de sacarose em palatinose, um açúcar alternativo que apresenta baixa cariogenicidade. Metodologia de Superfície de Resposta foi empregada com sucesso para determinar a concentração ótima dos componentes do meio de cultivo. A máxima produção de glicosiltransferase (21,78 U mL-1) foi obtida utilizando o meio de cultivo otimizado composto por melaço de cana de açúcar (80 g L-1), peptona bacteriológica (7 g L-1) e extrato de levedura (20 g L-1), após 8 horas de fermentação a 28°C. A conversão de sacarose em palatinose foi estudada utilizando células imobilizadas em alginato de cálcio. Os efeitos da concentração de alginato (2-4%), concentração de massa celular (20-40%) e concentração de substrato (25-45%) foram avaliados e a porcentagem de palatinose foi de aproximadamente 62,5%.