Exopolysaccharides produced by the symbiotic nitrogen-fixing bacteria of leguminosae

The process of biological nitrogen fixation (BNF), performed by symbiotic nitrogen fixing bacteria with legume species, commonly known as α and β rhizobia, provides high sustainability for the ecosystems. Its management as a biotechnology is well succeeded for improving crop yields. A remarkable example of this success is the inoculation of Brazilian soybeans with Bradyrhizobium strains. Rhizobia produce a wide diversity of chemical structures of exopolysaccharides (EPS). Although the role of EPS is relatively well studied in the process of BNF, their economic and environmental potential is not yet explored. These EPS are mostly species-specific heteropolysaccharides, which can vary according to the composition of sugars, their linkages in a single subunit, the repeating unit size and the degree of polymerization. Studies have showed that the EPS produced by rhizobia play an important role in the invasion process, infection threads formation, bacteroid and nodule development and plant defense response. These EPS also confer protection to these bacteria when exposed to environmental stresses. In general, strains of rhizobia that produce greater amounts of EPS are more tolerant to adverse conditions when compared with strains that produce less. Moreover, it is known that the EPS produced by microorganisms are widely used in various industrial activities. These compounds, also called biopolymers, provide a valid alternative for the commonly used in food industry through the development of products with identical properties or with better rheological characteristics, which can be used for new applications. The microbial EPS are also able to increase the adhesion of soil particles favoring the mechanical stability of aggregates, increasing levels of water retention and air flows in this environment. Due to the importance of EPS, in this review we discuss the role of these compounds in the process of BNF, in the adaptation of rhizobia to environmental stresses and in the process of soil aggregation. The possible applications of these biopolymers in industry are also discussed.

Characterization of Exopolysaccharides Produced by Rhizobia Species

ABSTRACT Increasing attention has been given, over the past decades, to the production of exopolysaccharides (EPS) from rhizobia, due to their various biotechnological applications. Overall characterization of biopolymers involves evaluation of their chemical, physical, and biological properties; this evaluation is a key factor in understanding their behavior in different environments, which enables researchers to foresee their potential applications. Our focus was to study the EPS produced by Mesorhizobium huakuii LMG14107, M. loti LMG6125, M. plurifarium LMG11892,Rhizobium giardini bv. giardiniH152T, R. mongolense LMG19141, andSinorhizobium (= Ensifer)kostiense LMG19227 in a RDM medium with glycerol as a carbon source. These biopolymers were isolated and characterized by reversed-phase high-performance liquid chromatography (RP-HPLC), Fourier transform infrared (FTIR), and nuclear magnetic resonance (NMR) spectroscopies. Maximum exopolysaccharide production was 3.10, 2.72, and 2.50 g L-1for the strains LMG6125, LMG19227, and LMG19141, respectively. The purified EPS revealed prominent functional reactive groups, such as hydroxyl and carboxylic, which correspond to a typical heteropolysaccharide. The EPS are composed primarily of galactose and glucose. Minor components found were rhamnose, glucuronic acid, and galacturonic acid. Indeed, from the results of techniques applied in this study, it can be noted that the EPS are species-specific heteropolysaccharide polymers composed of common sugars that are substituted by non-carbohydrate moieties. In addition, analysis of these results indicates that rhizobial EPS can be classified into five groups based on ester type, as determined from the 13C NMR spectra. Knowledge of the EPS composition now facilitates further investigations relating polysaccharide structure and dynamics to rheological properties.

An alternative method for screening lactic acid bacteria for the production of exopolysaccharides with rapid confirmation

The accumulation of exopolysaccharides (EPS) produced by microorganisms occurs in the presence of excess substrate and limiting conditions of elements that are essential to growth, such as nitrogen, phosphorus, sulfur, and magnesium. The presence of EPS produced by bacterial cells contributes to slime colonies formation in solid medium and increased viscosity in liquid medium. This paper proposes an alternative method for screening EPS-producing lactic acid bacteria using solid medium-containing discs of filter paper that are saturated with active cultures. The screening was carried out under different culture conditions varying the type of sugar, pH, and temperature. EPS production was visualized by the presence of mucoid colonies on the discs, which was confirmed by the formation of a precipitate when part of this colony was mixed with absolute alcohol. The established conditions for obtaining a high number of isolates producing EPS were 10% sucrose, pH 7.5 and 28 ºC. This method proved to be effective and economical because several strains could be tested on the same plate, with immediate confirmation.

Caracterização química dos géis produzidos pelas bactérias diazotróficas Rhizobium tropici e Mesorhizobium sp.

The exopolysaccharides with characteristics of gel produced by Rhizobium tropici (EPS RT) and Mesorhizobium sp (EPS MR) are acidic heteropolysaccharide composed mainly of glucose and galactose in a molar ratio of 4:1 and 5:1 respectively, with traces of mannose (~ 1%). Chemical analysis showed the presence of uronic acid, pyruvate and acetyl-substituents in the structures of both polymers. Experiments of gel permeation chromatography and polyacrylamide gel electrophoresis showed that EPS RT and EPS MR are homogeneous molecules with low grade of polydispersity. The EPS were characterized using spectroscopic techniques of FT-IR, ¹H and 13C-NMR.

Rheology of spreadable goat cheese made with autochthonous lactic cultures differing in their ability to produce exopolysaccharides

The aim of this study was to compare the rheology of spreadable cheeses elaborated with autochthonous lactic starter cultures without the addition of exopolysaccharide-producing strain in the same starter with exopolysaccharide-producing strain. From a rheological standpoint, both samples were characterized as weak viscoelastic gels and pseudoplastic products. It was concluded that cheese made with exopolysaccharide-producing strain showed smaller G', G", and η* values over the range of frequencies studied and smaller critic stress values than the cheese without exopolysaccharide-producing strain. The results obtained indicate that cheeses without exopolysaccharide-producing strain need to be added with any texture enhancer product.

Intra-His bundle block: clinical, electrocardiographic, and electrophysiologic characteristics

OBJECTIVE: To assess the clinical, electrocardiographic, and electrophysiologic characteristics of patients (pt) with intra-His bundle block undergoing an electrophysiologic study (EPS). METHODS: We analyzed the characteristics of 16 pt with second-degree atrioventricular block and symptoms of syncope or dyspnea, or both, undergoing conventional EPS. RESULTS: Intra-His bundle block was documented in 16 pt during an EPS. In 15 (94%) pt, the atrioventricular block was recorded in sinus rhythm; 4 (25%) pt had intra-His Wenckebach phenomenon, which correlated with Mobitz I (MI) atrioventricular block on the electrocardiogram. Seven (44%) pt had 2:1 atrioventricular block, 2 of whom were asymptomatic (12.5%). One (6%) pt had intra- and infra-His bundle block. Clinically, 11 (68%) pt had syncope or presyncope, 3 (18%) had dyspnea on exertion, and 2 (12.5%) were asymptomatic. Eight (50%) pt had bundle-branch block as follows: 4 (25%) pt had left bundle-branch block, and 4 (25%) had right bundle-branch block. Left anterosuperior divisional block was observed in 3 pt (19%), 2 of whom with associated right bundle-branch block. CONCLUSION: Intra-His bundle block was observed in 11% of the pt with second-degree atrioventricular block, syncope or presyncope, or both, it being the most frequent clinical presentation. Intra-His bundle block was more common in the elderly (> 60 years) and among females. The most frequent electrocardiographic presentations were second-degree Mobitz I or type 2:1 atrioventricular block.

Value of the Qrs-T Angle in Predicting the Induction of Ventricular Tachyarrhythmias in Patients with Chagas Disease

Background:The QRS-T angle correlates with prognosis in patients with heart failure and coronary artery disease, reflected by an increase in mortality proportional to an increase in the difference between the axes of the QRS complex and T wave in the frontal plane. The value of this correlation in patients with Chagas heart disease is currently unknown.Objective:Determine the correlation of the QRS-T angle and the risk of induction of ventricular tachycardia / ventricular fibrillation (VT / VF) during electrophysiological study (EPS) in patients with Chagas disease.Methods:Case-control study at a tertiary center. Patients without induction of VT / VF on EPS were used as controls. The QRS-T angle was categorized as normal (0-105º), borderline (105-135º) or abnormal (135-180º). Differences between groups for continuous variables were analyzed with the t test or Mann-Whitney test, and for categorical variables with Fisher's exact test. P values < 0.05 were considered significant.Results:Of 116 patients undergoing EPS, 37.9% were excluded due to incomplete information / inactive records or due to the impossibility to correctly calculate the QRS-T angle (presence of left bundle branch block and atrial fibrillation). Of 72 patients included in the study, 31 induced VT / VF on EPS. Of these, the QRS-T angle was normal in 41.9%, borderline in 12.9% and abnormal in 45.2%. Among patients without induction of VT / VF on EPS, the QRS-T angle was normal in 63.4%, borderline in 14.6% and abnormal in 17.1% (p = 0.04). When compared with patients with normal QRS-T angle, those with abnormal angle had a fourfold higher risk of inducing ventricular tachycardia / ventricular fibrillation on EPS [odds ratio (OR) 4; confidence interval (CI) 1.298-12.325; p = 0.028]. After adjustment for other variables such as age, ejection fraction (EF) and QRS size, there was a trend for the abnormal QRS-T angle to identify patients with increased risk of inducing VT / VF during EPS (OR 3.95; CI 0.99-15.82; p = 0.052). The EF also emerged as a predictor of induction of VT / VF: for each point increase in EF, there was a 4% reduction in the rate of sustained ventricular arrhythmia on EPS.Conclusions:Changes in the QRS-T angle and decreases in EF were associated with an increased risk of induction of VT / VF on EPS.

Managers' perspective on continuous health education in a region of São Paulo State

OBJECTIVEAnalysing the concepts of Continuous Health Education - CHE (EPS - in Portuguese), operated by municipal managers and translated into official documents.METHODQualitative research with the use of official documents and semi-structured interviews with the Municipal Health Secretaries or Coordinators of Primary Health Care in the Northeast Region of São Paulo State, and thematic analysis of empirical material.RESULTSResults indicate difficulties in the municipalities problematizing their management practices, services and health care; EPS tools presented are insufficient and unsatisfactory for amending the array of problems raised and are still far from the routine of Primary Care services.CONCLUSIONDespite efforts to implement EPS actions for the strengthening of primary care, the process appears to be incipient.

Assessment of soil properties by organic matter and EM-microorganism incorporation

Properties of a claim loam soil, collected in Aranjuez (Madrid) and enriched with organic matter and microorganisms, were evaluated under controlled temperature and moisture conditions, over a period of three months. The following treatments were carried out: soil (control); soil + 50 t ha-1 of animal manure (E50); soil + 50 t ha-1 of animal manure + 30 L ha-1 of effective microorganisms (E50EM); soil + 30 t ha-1 of the combination of various green crop residues and weeds (RC30) and soil + 30 t ha-1 of the combination of various green crop residues and weeds + 30 L ha-1 of effective microorganisms (RC30EM). Soil samples were taken before and after incubation and their physical, chemical, and microbiological parameters analyzed. Significant increase was observed in the production of exopolysaccharides and basic phosphatase and esterase enzyme activities in the treatments E50EM and RC30EM, in correlation with the humification of organic matter, water retention at field capacity, and the cationic exchange capacity (CEC) of the same treatments. The conclusion was drawn that the incorporation of a mixture of effective microorganisms (EM) intensified the biological soil activity and improved physical and chemical soil properties, contributing to a quick humification of fresh organic matter. These findings were illustrated by the microbiological activities of exopolysaccharides and by alkaline phosphatase and esterase enzymes, which can be used as early and integrated soil health indicators.

Establishment of an in vitro system for studies on the induced resistance of cotton to Xanthomonas campestris pv. malvacearum

An in vitro system for studying the resistance response of cotton (Gossypium hirsutum L.) to Xanthomonas campestris pv. malvacearum was investigated. Cell suspension cultures, established from hypocotyl-derived callus of cotton cultivar 101-102B, were treated with bacterial extracellular polysaccharides (EPS) extracted from the incompatible race 18 of X. campestris pv. malvacearum. EPS at 600 mug/mL caused pronounced darkening of the suspension cultures, as indicative of cell death, 48 hours after incubation. Protein electrophoresis analysis of the time course of EPS-treated cells showed differential accumulation of several protein bands after 12-24 hours. The time course of protein accumulation and cell death was consistent with an elicitor-mediated hypersensitive response.

Produção e comportamento reológico de exopolissacarídeos sintetizados por rizóbios isolados de guandu

O objetivo deste trabalho foi avaliar a produção e o comportamento reológico de exopolissacarídeos (EPS) produzidos por rizóbios isolados de nódulos de guandu [Cajanus cajan (L.) Millsp.], e a similaridade desses isolados, pela técnica de análise de restrição do DNA ribossomal amplificado. As bactérias foram cultivadas em meio YM líquido e os EPS foram obtidos por precipitação com etanol gelado a partir do caldo de cultivo. Em seguida, eles foram recuperados por centrifugação, secados a vácuo, pesados e ressuspensos em água para avaliações reológicas. Os três isolados avaliados apresentaram diferenças na produção e na eficiência relativa da produção de EPS, com destaque para o isolado 8.1c. Os EPS demonstraram comportamento não newtoniano e pseudoplástico, porém também apresentaram diferenças na viscosidade aparente em uma mesma taxa de cisalhamento. Na taxa de cisalhamento de 1 s-1, os três isolados foram diferentes, enquanto na taxa de cisalhamento de 40 s-1, os isolados 53.5 e 30.6a2 foram iguais e diferiram do isolado 8.1c. A similaridade dos isolados foi condizente com os resultados de reologia dos EPS, em que bactérias que sintetizaram EPS com menor viscosidade aparente apresentaram maior similaridade

Caracterização físico-química, bioquímica e funcional da jabuticaba armazenada sob diferentes temperaturas

O presente trabalho teve como objetivo avaliar a qualidade pós-colheita de jabuticabas submetidas a diferentes temperaturas de armazenamento refrigerado (AR). Após a colheita, os frutos fisiologicamente maduros foram acondicionados em bandejas de poliestireno expandido (EPS), revestidas por filme plástico de polietileno de baixa densidade (PEBD) e armazenados sob refrigeração a 0; 3; 6; 9 e 12 ± 1ºC e U.R. 87 ± 2%, sendo avaliados a cada 5 dias. Os frutos foram avaliados quanto à perda de massa, atividade respiratória, pH, acidez titulável, sólidos solúveis, ácido ascórbico, textura, pectina total e solúvel, atividade da enzima polifenoloxidase (PFO), compostos fenólicos e atividade antioxidante. Para frutos refrigerados a 9 e 12ºC, o pico respiratório atrasou em relação aos demais tratamentos, além de apresentarem as menores taxas respiratórias. O teor de sólidos solúveis aumentou com o tempo de armazenamento para todas as temperaturas, contudo, em 9 e 12ºC, esse aumento foi em menor proporção. A firmeza e o teor de ácido ascórbico também foram superiores nos frutos armazenados a 9 e 12ºC, enquanto os contéudos de pectina solúvel foram menores. Observou-se a diminuição da atividade da enzima PFO ao longo dos 30 dias do AR, independentemente da temperatura utilizada; entretanto, os menores valores foram encontrados nos frutos mantidos a 9 e 12ºC. Os frutos armazenados a 12ºC apresentaram os maiores conteúdos de compostos fenólicos totais e a maior atividade antioxidante ao final do experimento. Nesse sentido, a temperatura de 12ºC foi a mais efetiva na manutenção da qualidade pós-colheita das jabuticabas.

Radiação gama na conservação de caqui 'Giombo', destanizado e frigoarmazenado

O caqui apresenta uma safra curta, sendo necessário estender seu período de comercialização utilizando técnicas adequadas de armazenamento. O objetivo deste trabalho foi avaliar o uso da radiação gama na qualidade pós-colheita de caquis 'Giombo' destanizados. Frutos colhidos meio-maduros e com aproximadamente 50 % da coloração verde, foram destanizados, acondicionados em bandejas de poliestireno expandido (EPS), revestidas por filme plástico de polietileno de baixa densidade (PEBD) e submetidos à radiação gama (60Co). Os frutos foram armazenados durante 35 dias, sob refrigeração (0±0,5 ºC e 85 ± 0,5% UR). Os tratamentos realizados foram: T1 - 0,0 kGy; T2 - 0,3 kGy; T3 - 0,6 kGy; T4 - 0,9 kGy; T5 - 1,2 kGy. As análises realizadas a cada 7 dias foram: perda de massa, atividade respiratória, teores de sólidos solúveis (SS), ácido ascórbico, acidez titulável (AT) e relação SS/AT. Os frutos submetidos à dose de 0,6 kGy apresentaram o menor percentual de perda de massa e produção de CO2 ao longo do período experimental. Os teores de SS e AT permaneceram estáveis e sem diferenças devido às doses de irradiação aplicadas. Os tratamentos com 0,3 KGy e 0,6 KGy foram os mais eficazes na manutenção da firmeza nos caquis. Nos teores de ácido ascórbico, observou-se redução ao longo do período experimental, sendo os menores valores apresentados para a maior dose de irradiação (1,2 KGy).

Caracterização química de glucanas fúngicas e suas aplicações biotecnológicas

This article gives an overview of exopolysaccharides produced by fungi. The structural characterization of beta-D-(1->3) and beta-D-(1->3, 1->6)- glucans is discussed focusing on different chemical and physical procedures. The industrial applications are also considered mainly from the point of view of human health.

Purificação e caracterização de uma gentioexaose obtida de botriosferana por hidrólise ácida parcial

A hexa-oligosaccharide was obtained by partial acid hydrolysis from botryosphaeran, an exopolysaccharide (EPS) β(1→3; 1→6)-D-glucan type, produced by the ascomyceteous fungus Botryosphaeria rhodina. The oligosaccharide was purified by gel filtration and charcoal-Celite column chromatography and the analysis was followed by HPAEC/PAD. The structure was determined by NMR spectroscopy and mass spectrometry, which showed that the oligosaccharide consists of six β-D-glucopiranosyl units O-6 substituted (gentiohexaose).