Syntrophic relationships between algae and bacteria in a fresh-water stream and in culture /

Interactions between freshwater algae and bacteria were examined in a natural stream habitat and a laboratory model. Field observations provided circumstantial evidence, in statistical correlation for syntrophy between the microbial populations. This relation is probably subject to control by the temperature and pH of the aquatic environment. Several species of a pond community were isolated in axenic culture and tests were performed to determine the nature of mixed species interactions. Isolation procedures and field studies indicated that selected strains of Chlorella and Azotobacter were closely associated in their natural habitat. With the suspected controlling parameters, pH and temperature, held constant, mixed cultures of algae and bacteria were compared to axenic cultures of the same organisms, and a mutual stimulation of growth was observed. A mixed pure culture apparatus was designed in this laboratory to study the algal-bacterial interaction and to test the hypothesis that such an interaction may take place through a diffusable substance or through certain medium-borne conditions, Azotobacter was found to take up a Chlorella-produced exudate, to stimulate protein synthesis, to enhance chlorophyll production and to cause a numerical increase in the interacting Chlorella population. It is not clear whether control is at the environmental, cellular or genetic level in these mixed population interactions. Experimental observations in the model system, taken with field correlations allow one to state that there may be a direct relationship governing the population fluctuations of these two organisms in their natural stream surroundings.

Process Improvement of Coir Pith Degradation Involving White Rot Fungi By Substitution of Urea With Nitrogen Fixing Bacteria and Application of Biodegraded Coir Pith as Plant Growth Medium

In this study, a novel improved technology could be developed to convert the recalcitrant coir pith into environmental friendly organic manure. The standard method of composting involves the substitution of urea with nitrogen fixing bacteria viz. Azotobacter vinelandii and Azospirillum brasilense leading to the development of an improved method of coir pith. The combined action of the microorganisms could enhance the biodegradation of coir pith. In the present study, Pleurotus sajor caju, an edible mushroom which has the ability to degrade coir pith, and the addition of nitrogen fixing bacteria like Azotobacter vinelandii and Azospirillum brasilense could accelerate the action of the fungi on coir pith. The use of these microorganisms brings about definite changes in the NPK, Ammonia, Organic Carbon and Lignin contents in coir pith. This study will encourage the use of biodegraded coir pith as organic manure for agri/horti purpose to get better yields and can serve as a better technology to solve the problem of accumulated coir pith in coir based industries

Ciclo del nitrógeno : prácticas de microbiología del suelo.

Se expone una práctica sencilla de microbiología para la iniciación de los alumnos de bachillerato, que requiere medios relativamente sencillos. Se trata del estudio de la fijación del nitrógeno atmosférico, mediante el reconocimiento de un fijador libre del suelo, Azotobacter, y a dos simbióticos, Rhizobium y Frankia.

Alginato bacteriano: aspectos tecnológicos, características e produção

Alginate is a biopolymer used for a variety of industrial applications, for example, in the textiles, cosmetics, foods, agricultural and biotechnological industries. This biopolymer is traditionally extracted from some brown seaweeds (Phaeophyceae) and can be produced by bacteria isolated from soil, as Azotobacter vinelandii, like capsular polysaccharide using glucose, sucrose, among others as carbon sources. The main difference between the alginate of seaweed and the bacterial ones, is the biggest degree of acetylation of this last one, with great influence in the gel force. These chemical characteristics and production of bacterial alginate are presented in this work.

Processo biotecnologico a cellule immobilizzate per la produzione di acidi grassi volatili da acque di vegetazione

Le acque di vegetazione (AV) costituiscono un serio problema di carattere ambientale, sia a causa della loro elevata produzione sia per l’ elevato contenuto di COD che oscilla fra 50 e 150 g/l. Le AV sono considerate un refluo a tasso inquinante fra i più elevati nell’ambito dell’industria agroalimentare e la loro tossicità è determinata in massima parte dalla componente fenolica. Il presente lavoro si propone di studiare e ottimizzare un processo non solo di smaltimento di tale refluo ma anche di una sua valorizzazione, utlizzandolo come materia prima per la produzione di acidi grassi e quindi di PHA, polimeri biodegradabili utilizzabili in varie applicazioni. A tale scopo sono stati utilizzati due bioreattori anaerobici a biomassa adesa, di identica configurazione, con cui si sono condotti due esperimenti in continuo a diverse temperature e carichi organici al fine di studiare l’influenza di tali parametri sul processo. Il primo esperimento è stato condotto a 35°C e carico organico pari a 12,39 g/Ld, il secondo a 25°C e carico organico pari a 8,40 g/Ld. Si è scelto di allestire e mettere in opera un processo a cellule immobilizzate in quanto questa tecnologia si è rivelata vantaggiosa nel trattamento continuo di reflui ad alto contenuto di COD e carichi variabili. Inoltre si è scelto di lavorare in continuo poiché tale condizione, per debiti tempi di ritenzione idraulica, consente di minimizzare la metanogenesi, mediata da microrganismi con basse velocità specifiche di crescita. Per costituire il letto fisso dei due reattori si sono utilizzati due diversi tipi di supporto, in modo da poter studiare anche l’influenza di tale parametro, in particolare si è fatto uso di carbone attivo granulare (GAC) e filtri ceramici Vukopor S10 (VS). Confrontando i risultati si è visto che la massima quantità di VFA prodotta nell’ambito del presente studio si ha nel VS mantenuto a 25°C: in tale condizione si arriva infatti ad un valore di VFA prodotti pari a 524,668 mgCOD/L. Inoltre l’effluente in uscita risulta più concentrato in termini di VFA rispetto a quello in entrata: nell’alimentazione la percentuale di materiale organico presente sottoforma di acidi grassi volatili era del 54 % e tale percentuale, in uscita dai reattori, ha raggiunto il 59 %. Il VS25 rappresenta anche la condizione in cui il COD degradato si è trasformato in percentuale minore a metano (2,35 %) e questo a prova del fatto che l’acidogenesi ha prevalso sulla metanogenesi. Anche nella condizione più favorevole alla produzione di VFA però, si è riusciti ad ottenere una loro concentrazione in uscita (3,43 g/L) inferiore rispetto a quella di tentativo (8,5 g/L di VFA) per il processo di produzione di PHA, sviluppato da un gruppo di ricerca dell’università “La Sapienza” di Roma, relativa ad un medium sintetico. Si può constatare che la modesta produzione di VFA non è dovuta all’eccessiva degradazione del COD, essendo questa nel VS25 appena pari al 6,23%, ma piuttosto è dovuta a una scarsa concentrazione di VFA in uscita. Questo è di buon auspicio nell’ottica di ottimizzare il processo migliorandone le prestazioni, poiché è possibile aumentare tale concentrazione aumentando la conversione di COD in VFA che nel VS25 è pari a solo 5,87%. Per aumentare tale valore si può agire su vari parametri, quali la temperatura e il carico organico. Si è visto che il processo di acidogenesi è favorito, per il VS, per basse temperature e alti carichi organici. Per quanto riguarda il reattore impaccato con carbone attivo la produzione di VFA è molto ridotta per tutti i valori di temperatura e carichi organici utilizzati. Si può quindi pensare a un’applicazione diversa di tale tipo di reattore, ad esempio per la produzione di metano e quindi di energia.

Calidad poscosecha del tomate (Solanum lycopersicum L.) cultivado en sistemas ecológicos de fertilización

Se condujeron cuatro experimentos unifactoriales en condiciones de campo en áreas agrícolas de la Universidad de Granma, en el período comprendido de septiembre/2007 a enero/2010 para determinar el efecto de la aplicación de micorrizas arbusculares, Azotobacter chroococcum y ácidos húmicos sobre la productividad y la calidad poscosecha del tomate (Solanum lycopersicum L.) cv. “Vyta”, sobre un suelo de tipo Fluvisol. La aplicación simple y combinada de las micorrizas arbusculares, el Azotobacter chroococcum y los ácidos húmicos mostró un efecto positivo sobre la productividad de la variedad de tomate estudiada, al obtenerse incrementos significativos de este indicador en comparación con el control (sin aplicación), lográndose los mejores resultados con la combinación Glomus fasciculatum (0,3 kg m-2) + Azotobacter chroococcum (30 L ha-1) + ácidos húmicos (500 mg L-1) con valores de 54,57 t ha-1. Se comprobó además, que los tratamientos con estos bioproductos presentaron contenidos de sólidos solubles totales (SST), carbohidratos solubles totales (CST), vitamina C, porcentaje de materia seca (PMS), firmeza, longitud y anchura de los frutos superiores al control (sin aplicación). Las plantas controles (sin aplicación) significativamente incrementaron la acidez titulable (contenido de ácido cítrico) y la pérdida de peso y redujeron el pH de los frutos del tomate. Los análisis de correlación realizados revelan que existieron correlaciones positivas y significativas entre el contenido de vitamina C, materia seca, sólidos solubles totales, carbohidratos solubles totales, y el pH de los frutos, mientras que la acidez titulable presentó correlaciones negativas y significativas con todas las variables mencionadas anteriormente. Se encontró a su vez un efecto sinérgico en las plantas con la aplicación combinada de estos productos con respecto al control (sin aplicación), expresado en los mayores valores de los parámetros micorrízicos estudiados. ABSTRACT Four unifactorial experiments under field conditions in an agricultural area belonging to the University of Granma from September/2007 to January/2010 aiming to evaluate the effect of arbuscular mycorrhizas, Azotobacter and humic acids on tomato (Solanum lycopersicum L) cv. “Vyta” crop yield and postharvest quality in a Typic Fluvent soil were conducted. The simple and mixed application of arbuscular mycorrhizas, Azotobacter chroococcum and humic acids showed a positive effect on yield and internal postharvest parameters evaluated for the studied tomato variety, achieving significant increases with respect to the control treatment ones. The best results related to crop yield were achieved by combining Glomus fasciculatum (0.3 kg m-2) + Azotobacter chroococcum (30 L ha-1) + humic acids (500 mg L-1) that lead to 54.57 t ha-1. In addition, it was also demonstrated that the treatments under effect of biofertilizers and biostimulants resulted in higher total soluble solids (TSS), total soluble carbohydrates (TSC), vitamin C, dry matter contents (DMC), firmness, length and width of the fruits than the control treatment. The control plants showed significantly increased titrable acidity (citric acid content) and weight loss and reduced pH on the tomato fruits. The conducted correlation analyses revealed that there were significant and positive correlations between vitamin C content, dry matter, total soluble solids, total soluble carbohydrates, and pH of the fruit, while titratable acidity showed significant negative correlations with all variables mentioned above. At the same time, a synergic effect by mixed utilization of these biological products on tomato plants, with respect to control treatment (non-treated), was found regarding the increase of studied mycorrhizal parameters.

Pseudomonas aeruginosa killing of Caenorhabditis elegans used to identify P. aeruginosa virulence factors

We reported recently that the human opportunistic pathogen Pseudomonas aeruginosa strain PA14 kills Caenorhabditis elegans and that many P. aeruginosa virulence factors (genes) required for maximum virulence in mouse pathogenicity are also required for maximum killing of C. elegans. Here we report that among eight P. aeruginosa PA14 TnphoA mutants isolated that exhibited reduced killing of C. elegans, at least five also exhibited reduced virulence in mice. Three of the TnphoA mutants corresponded to the known virulence-related genes lasR, gacA, and lemA. Three of the mutants corresponded to known genes (aefA from Escherichia coli, pstP from Azotobacter vinelandii, and mtrR from Neisseria gonorrhoeae) that had not been shown previously to play a role in pathogenesis, and two of the mutants contained TnphoA inserted into novel sequences. These data indicate that the killing of C. elegans by P. aeruginosa can be exploited to identify novel P. aeruginosa virulence factors important for mammalian pathogenesis.

The chaperone GroEL is required for the final assembly of the molybdenum-iron protein of nitrogenase

It is known that an E146D site-directed variant of the Azotobacter vinelandii iron protein (Fe protein) is specifically defective in its ability to participate in iron-molybdenum cofactor (FeMoco) insertion. Molybdenum-iron protein (MoFe protein) from the strain expressing the E146D Fe protein is partially (≈45%) FeMoco deficient. The “free” FeMoco that is not inserted accumulates in the cell. We were able to insert this “free” FeMoco into the partially pure FeMoco-deficient MoFe protein. This insertion reaction required crude extract of the ΔnifHDK A. vinelandii strain CA12, Fe protein and MgATP. We used this as an assay to purify a required “insertion” protein. The purified protein was identified as GroEL, based on the molecular mass of its subunit (58.8 kDa), crossreaction with commercially available antibodies raised against E. coli GroEL, and its NH2-terminal polypeptide sequence. The NH2-terminal polypeptide sequence showed identity of up to 84% to GroEL from various organisms. Purified GroEL of A. vinelandii alone or in combination with MgATP and Fe protein did not support the FeMoco insertion into pure FeMoco-deficient MoFe protein, suggesting that there are still other proteins and/or factors missing. By using GroEL-containing extracts from a ΔnifHDK strain of A. vinelandii CA12 along with FeMoco, Fe protein, and MgATP, we were able to supply all required proteins and/or factors and obtained a fully active reconstituted E146D nifH MoFe protein. The involvement of the molecular chaperone GroEL in the insertion of a metal cluster into an apoprotein may have broad implications for the maturation of other metalloenzymes.

Statnote 29:discriminant analysis

Discriminant analysis (also known as discriminant function analysis or multiple discriminant analysis) is a multivariate statistical method of testing the degree to which two or more populations may overlap with each other. It was devised independently by several statisticians including Fisher, Mahalanobis, and Hotelling ). The technique has several possible applications in Microbiology. First, in a clinical microbiological setting, if two different infectious diseases were defined by a number of clinical and pathological variables, it may be useful to decide which measurements were the most effective at distinguishing between the two diseases. Second, in an environmental microbiological setting, the technique could be used to study the relationships between different populations, e.g., to what extent do the properties of soils in which the bacterium Azotobacter is found differ from those in which it is absent? Third, the method can be used as a multivariate ‘t’ test , i.e., given a number of related measurements on two groups, the analysis can provide a single test of the hypothesis that the two populations have the same means for all the variables studied. This statnote describes one of the most popular applications of discriminant analysis in identifying the descriptive variables that can distinguish between two populations.

Statnote 32 : the partial correlation coefficient

In previous statnotes, the application of correlation and regression methods to the analysis of two variables (X,Y) was described. The most important statistic used to measure the degree of correlation between two variables is Pearson’s ‘product moment correlation coefficient’ (‘r’). The correlation between two variables may be due to their common relation to other variables. Hence, investigators using correlation studies need to be alert to the possibilities of spurious correlation and the methods of ‘partial correlation’ are one method of taking this into account. This statnote applies the methods of partial correlation to three scenarios. First, to a fairly obvious example of a spurious correlation resulting from the ‘size effect’ involving the relationship between the number of general practitioners (GP) and the number of deaths of patients in a town. Second, to the relationship between the abundance of the nitrogen-fixing bacterium Azotobacter in soil and three soil variables, and finally, to a more complex scenario, first introduced in Statnote 24involving the relationship between the growth of lichens in the field and climate.