Enhancing Bio-Available Phosphorous in Soil Through Sulfur Oxidation by Thiobacilli.

Aims: The objectives were to evaluate the phosphate solubilization efficiency of different Thiobacilli strains and to find out the best combination of sulfur and Thiobacilli for enhancing bio-available P in soil. Study Design: An experimental study. Place and Duration of Study: Microbiology and Soil Fertility Labs, Department of Soil Science and Soil and Water Conservation, Pir Mehr Ali Shah Arid Agriculture University, Rawalpindi, Pakistan and Microbiology and Soil Chemistry Labs, Auriga Research Center, Lahore, Pakistan, between May 2011 and November 2012. Methodology: Fifty Thiobacilli strains were isolated from ten different ecologies. Then an incubation study of soil was performed wherein the most efficient four Thiobacilli strains were inoculated in combination with three different levels of elemental sulfur to determine pH, water soluble sulfur, sequential P fractions and bio-available phosphorous contents in the incubated soil. Results: All the four Thiobacillus strains (IW16, SW2, IW1 and IW14) dropped pH of the incubated soil along with three doses of S° (50, 75 and 100 kg ha-1). However, Thiobacillus strains IW16 and SW2 reduced soil pH quite sharply from 7.90 to 7.12 (net reduction of 0.78 points) and 7.28 (net reduction of 0.62 points) respectively where inoculated with S° @ 100 kg ha-1. The best P solubilizer was Thiobacillus strain IW16 and the best dose of S° was @ 100 kg ha-1 and their combination enhanced maximum quantity of P (22.26 mg kg-1) in the soil by solubilizing already present insoluble calcium bounded P fractions like octacalcium phosphate (Ca8-P) and apatite (Ca10-P). Conclusion: The present study suggests the use of Thiobacilli along with elemental sulfur for the dissolution and enhancement of bio-available P in alkaline and calcareous soils.

Modelling of the biofiltration of reduced sulphur compounds through biotrickling filters connected in series: effect of H2S

Background: The behaviour of two biotrickling filters connected in serie (BTF) inoculated with Acidithiobacillus thiooxidans and Thiobacillus thioparus, biodegrading hydrogen sulphide (H2S) and dimethyl sulphide (DMS) simultaneously were studied. A model which considers gas to liquid mass transfer and biooxidation in the biofilm attached to the support is developed. Additionally, a fixed bed biotrickling filter where the microorganism is immobilized in a biofilm which degrades a mixture of H2S and DMS is implemented. Validation of the model was carried out using experimental data obtained at different H2S and DMS loads. Results: The inhibitory effect caused by the presence of H2S on the DMS is observed, which is evidenced by the decrease of the DMS removal efficiency from 80 to 27 percent, due to the preference that T. thioparus has by simple metabolism. H2S is not affected by the DMS, with removal efficiencies of 95 to 97 percent, but it decreases at high concentrations of the compound, due to the inhibition of metabolism by high H2S input loads. The model which describes the BFT fits successfully with the experimental results and it has a high sensitivity to inhibition parameters. Conclusion: It is shown that the microorganism has a high affinity for H2S, producing substrate inhibition when the concentration is high. The H2S is able to inhibit the DMS biooxidation, whereas the DMS does not affect the H2S biooxidation.

Thiosulphate oxidation by Thiobacillus thioparus and Halothiobacillus neapolitanus strains isolated from the petrochemical industry

Sulphur Oxidizing Bacteria (SOB) is a group of microorganisms widely used for the biofiltration of Total Reduced Sulphur compounds (TRS). TRS are bad smelling compounds with neurotoxic activity which are produced by different industries (cellulose, petrochemical). Thiobacillus thioparus has the capability to oxidize organic TRS, and strains of this bacterium are commonly used for TRS biofiltration technology. In this study, two thiosulphate oxidizing strains were isolated from a petrochemical plant (ENAP BioBio, Chile). They were subjected to molecular analysis by real time PCR using specific primers for T. thioparus. rDNA16S were sequenced using universal primers and their corresponding thiosulphate activities were compared with the reference strain T. thioparus ATCC 10801 in batch standard conditions. Real time PCR and 16S rDNA sequencing showed that one of the isolated strains belonged to the Thiobacillus branch. This strain degrades thiosulphate with a similar activity profile to that shown by the ATCC 10801 strain, but with less growth, making it useful in biofiltration.

Performance of a biotrickling filter employing Thiobacillus thioparus immobilized on polyurethane foam for hydrogen sulfide removal

The removal of hydrogen sulfide [H[2]S] from contaminated airstream was studied in a biotrickling filter [BTF] packed with open-pore polyurethane foam as a carrier of Thiobacillus thioparus [DSMZ5368] with counter current gas/liquid flows. The effect of operating parameters on BTF performance was studied. Experiments were performed at different Empty Bed Residence Times [EBRT] from 9 to 45 seconds, and different initial H[2]S concentration from 25 to 85 ppm. The results showed reasonable performance of the BTF, in H[2]S removal from the synthetic gas stream. However, the performance was somewhat lower than other studies in BTF in which either Thiobacillus thioparus with other packings or polyurethane foam with other microbial cultures were used. The effect of liquid recirculation rate [LRR] in the range of 175-525 ml/min [0.46-1.34 m/h] on BTF performance was also studied. Results showed that increasing LRR from 175 to 350 mL/min resulted in significant enhancement of H2S removal efficiency, but further increase in LRR up to 525 mL/min had an insignificant effect. H[2]S elimination at different heights of the bed was studied and it was found that decrease in EBRT results in more homogeneous removal of the pollutant in BTF. Determination of microbial species in the BTF after 100 days performance showed that during BTF operation the only H[2]S degrading specie was Thiobacillus thioparus

Oxidation of volatile reduced sulphur compounds in biotrickling filter inoculated with Thiobacillus thioparus

Reduced volatile sulphur compounds generate an impact on the environment, because of the bad smell and its low odour threshold. Compared with the existing physicochemical technologies for their elimination, biotrickling filters are an economically and environmentally sustainable alternative. Usually mixed cultures of microorganisms are used for inoculating biotrickling filters, in this case a pure culture of Thiobacillus thioparus is used for generating a biofilm, allowing to measure its capacity for the oxidation of four volatile reduced sulphur compounds: hydrogen sulphide, dimethyl sulphide, methyl mercaptan and dimethyl disulphide, using a residence time of 0.033 hrs. The viable cells of the biofilm were quantified by epifluorescence microscopy, staining the cells with ethidium bromide and acridine orange, polymerase chain reaction analysis in real time was used for testing the predominance of T. thiopharus in the biofilm. The microorganism was able to adhere and grow on the surface of rings made of polyethylene, with a viable population of 7•10(7) cell•ring-1, a 74 percent of total cells. The real time PCR showed a persistence of the population of T. thioparus for more than 300 days of operation, without being displaced by other microbial species. The maximum elimination capacities for each compound were 34.4; 21.8; 30.8 and 25.6 gS•m-3•h-1 for H2S, dimethylsulphide, dimethyldisulphide and methyl mercaptan, respectively. We conclude that it is possible to implement a biotrickling filter with the bacteria T. thioparus, which can oxidize volatile reduced sulphur compounds efficiently.

Selection of sulfur oxidizing bacterium for sulfide removal in sulfate rich wastewater to enhance biogas production

Sulfur oxidizing bacteria (SOB) were isolated and tested in order to remove sulfide from high sulfate wastewater to reduce the amount of hydrogen sulfide (H2S) in the produced biogas. A promising SOB isolate, designated as isolate T307, was selected due to its best sulfide removal (86.7 percent) in the effluent of a sulfate reduction reactor (SRR) over a 24 hrs incubation. The bacterium was able to grow better as a mixotroph (yeast extract as a carbon source) than as a chemolithoautotroph. In addition, as a heterotroph, the bacterium grew well with yeast extract and peptone. Based on partial 16S rRNA gene sequence, the isolated T307 was an Alcaligenes sp. and was able to convert most of sulfide species (total sulfide: TS; dissolved sulfide: DS and H2S) into elemental sulfur or sulfate over a 20 hrs period of cultivation by controlling the speed of shaking. In a biogas reactor set, after pre-treating a sulfide medium with Alcaligenes sp. T307 there was a much higher specific yield of CH4 (238 ml CH4 g-1COD removed) and more biogas (154 ml L-1 d-1) was produced with the biogas containing more methane (48.1 percent CH4, 51.5 percent CO2 and 0.41 percent H2S) in comparison to a control with a specific yield of CH4, (72 ml CH4 g-1COD removed) 86 ml L-1 d-1 biogas produced with a composition of 35.5 percent CH4, 63.7 percent CO2 and 0.86 percent H2S.

Comparison on the removal of hydrogen sulfide in biotrickling filters inoculated with Thiobacillus thioparus and Acidithiobacillus thiooxidans

Emissions of hydrogen sulfide (H2S) by industrial activities is frequent cause of corrosion and unpleasant odours. Treatment of gaseous emissions contaminated with H2S by biotrickling filters inoculated with single cultures of sulfur oxidizer bacteria exhibit several advantages over physicochemical methods, such as shorter adaptation times and higher removal ability. Biofilms of Thiobacillus thioparus and Acidithiobacillus thiooxidans have proved to exhibit high removal capacities, yet no comparative studies between them have been reported. This article reports the efficiency of biotrickling filters inoculated with T. thioparus and A. thiooxidans under similar conditions excepting the pH, that was the optimal for the bacterial growth, for the removal of H2S. The support was selected by determining the respirometric coefficients of the biomass. The maximum removal capacity of the biofilter inoculated with T. thioparus, operating within the range of pH (5.5-7.0) was 14 gS m-3 h-1, lower the value obtained for the biotrickling filter inoculated with A. thiooxidans; 370 gS m-3 h-1. Therefore, it is concluded that acid biotrickling filter inoculated with A. thiooxidans constitute the best strategy to remove H2S, with the advantage that the system not require an exhaustive pH control of the liquid media.

Isolation of Thiobacillus sp from aerobic sludge of distillery and dairy effluent treatment plants and its sulfide oxidation activity at different concentrations.

In the present study two strains of Thiobacillus sp were isolated from aerobic sludge of distillery and dairy effluent treatment plant using standard methods of isolation and enrichment. Experiments were conducted using isolated cultures in batch bioreactor with initial sulfide concentration of 75 and 150 mg/l. The effect of initial sulfide concentration on the activity of isolated Thiobacillus sp was studied. Sulfide oxidizing capacity was also determined at different initial sulfide concentrations. The results from the study indicate the possible isolation of Thiobacillus cultures from native source and application in the full-scale reactor

Estudio piloto de aislamiento y fenotipificación de bacterias que participan en los procesos de biolixiviación, en las zonas mineras del Departamento de Boyaca / Pilot study on isolation and fenotipification of bacteria that participate in the process of biolixiviation, in the mining zones of the Department of Boyaca

Una alternativa para disminuir la contaminación ambiental generada por el empleo de sustancias químicas cuando se extraen metales pesados en la industria minera, así como los costos en materiales químicos es utilizar en este proceso microorganismos que han demostrado realizar esta actividad, denominados biolixiviadores. Para ello es necesario identificar y fenotipificar estos microorganismos. El objetivo de este proyecto fueaislar y fenotipificar microorganismos biolixiviadores obtenidos de cuatro minas de esmeraldas del departamentode Boyacá (Muzo, Coscuez, Chivor y Consorcio). El muestreo se realizó en agua y roca, habitat principal de las bacterias que participan en los procesos de biolixiviación. El aislamiento se realizó previapreparación de cada una de las muestras, las cuales se sembraron en el medio de cultivo 9K, líquido y sólido,e incubaron por siete días a 37°C. Se evidenciaron cambios físicos en la intensidad de color y en el aspecto en los medios de cultivo, lo que permite presumir que ha ocurrido alguna acción de las bacterias lixiviantes sobrelos compuestos del medio. La observación microscópica evidenció la presencia de bacilos gram negativos ycristales. Por otra parte, se confirmó la característica de ser bacterias acidófilas ya que su desarrollo seobtuvo a pH entre 1 y 2,5. Los resultados obtenidos en este estudio permitieron concluir que en estas minashay presencia de organismos bacterianos con morfología y comportamiento bioquímico compatible con bacteriasbiolixiviantes del género Thiobacillus. El aislamiento y fenotipificación obtenidos in vitro abre la posibilidadde establecer un método óptimo para su conservación y posterior utilización en procesos de bioremediación.

La incorporación de metales pesados en las bacterias y su importancia para el ambiente / The incorporation of heavy metal in the bactery and its importance for the environmental

Los metales pesados se han identificado como una clase reciente de contaminantes. Las bacterias por su parte, desempeñan un importantísimo papel como organismos modificadores de los ecosistemas, Esta revisión busca exclarecer los mecanismos de entrada de metales en las bacterias y la importancia toxicológica de dichos metales. Las interacciones bacteria-metal son conocidas y pueden ocurrir extracelularmente, en la superficie bacteriana, ó a nivel intracelular. Extracelularmente, se conocen el papel que desempeñan los microorganismos en la movilización de metales y la secreción de compuestos orgánicos de bajo peso molecular con alta afinidad por los metales conocidos como sideróforos. Las inteacciones de un metal dado con la superficie celular dependen del tipo de bacteria, ya que el metal interactúa con los grupos específicos cargados negativamente en cada una de ellas. A nivel intracelular, como consecuencia de la acumulación del metal, ocurren transformaciones enzimáticas ó la síntesis de proteínas específicas (metalotioninas). En general, las interacciones bacteria-metal se producen por una serie de eventos que se inician fuera del entorno celular, y finalizan intracelularmente. Estos eventos pueden ocurrir en secuencia o no, y las características de cada uno de ellos depende de la bacteria considerada, el compartimiento en el cual ocurre, y el tipo y especie química del metal

Modification of radiosensitivity of chlorpromazine with biological metal ions in Thiobacillus ferrooxidans.

Effect of chlorpromazine with biological metal ions, viz. calcium, magnesium, zink and copper was studied on T. ferrooxidans cell system. Chlorpromazine, calcium and magnesium alone could produce radioprotection. Maximum radioprotection was exhibited by chlorpromazine at lower concentration while copper and zink offered radiosensitization. However, combination of chlorpromazine with all biological metal ions exhibited radiosensitization. Dose modifying factor by chlorpromazine at lower concentration (0.025 mM) was 0.754 while in combination with Ca2+, Mg2+, Cu2+ and Zn2+ was 1.08, 1.25, 1.37 and 1.389 respectively. The possible interaction between chlorpromazine and biological metal ions is discussed at cellular membrane level.

The possible role of surface charge in membrane organization in an acidophile.

Thiobacillus ferrooxidans, an obligate acidophile, possesses an electron transport chain that uses oxidation of ferrous iron to generate proton motive force. The cells possess an efficient machinery for inter-conversion of the two components of the free energy, namely pH gradient and membrane potential. Incidentally, unlike most of the naturally occurring membranes, surface in T. ferrooxidans appears to be positively charged at or near its physiological pH. Independent estimate of such charge was obtained from binding studies with anionic optical and fluorescent probes. When the external pH is lowered below a critical value the inter-conversion of the two components of the proton motive force was no longer operative. A surface charge mediated phase separation of membrane is suggested as one of the possible mechanisms for the failure of such inter-conversion process.

Evidência sugerindo dois mecanismos de resistência ao arsenato em Thiobacillus ferrooxidans / Evidence for two arsenate resistance mechanisms in Thiobacillus ferrooxidans

Thiobacillus ferrooxidans é capaz de crescer na presença dos altos níveis de arsênio livre que säo gerados durante a recuperaçäo de ouro a partir de arsenopirita por lixiviaçäo. Os mecanismos dessa resistência näo säo conhecidos nessa bactéria. Uma cepa desta bactéria, isolada na Mina Morro Velho (Nova Lima, MG), foi cultivada em meio inorgânico 9K com concentraçöes de 0, 2, 4, 6 ou 8g.L-1 de arsenato de sódio em culturas semi-contínuas em fermentador de 10 litros. O crescimento celular foi pouco alterado por teores de arsenato inferiores a 4g.L-1. Para essa concentraçäo, o tempo de cultura foi consideralmente prolongado em funçäo de uma fase latente de cerca de 30h. Quando níveis de arsenato entre 4g.L-1 e 8g.L-1 foram utilizados, o perfise de cresciemtno das células adaptadas foi melhor. Este fenômeno näo foi observado nas culturas em frascos de 500mL (velocidade de agitaçäo: 150rpm). A comparaçäo dos perfis eletroforéticos (PAGE-SDS) dos extratos livres de células obtidos das bactérias cultuvadas sem ou na presença de 8g.L-1 de arsenato destacou, no último caso, quatro bandas maiores com peso molecular aparente de cerca de 16.500, 22.500, 24.000 e 33.500. Da, respectivamente. Esses resultados sugerem que há dois mecanismos de resistência em T. ferrooxidans para baixo e alto níveis de arsenato, respectivamente.

Role of rusticyanin in the electron transport process in Thiobacillus ferrooxidans.

Effect of diethyl dithiocarbamate (DEDC), an antimicrobial agent, on growth of Thiobacillus ferrooxidans, possibly by inhibiting rusticyanin present in the periplasmic space of the microorganism, has been studied to gain more insight into the electron transport chain in the bioleaching process. DEDC is found to form a stable complex with rusticyanin in solution and also in polyacrylamide gel. The spectrum of the complex is identical to that of Cu-DEDC complex, suggesting binding of DEDC with copper moiety of rusticyanin and resulting in inhibition of growth. In vitro reduction of purified rusticyanin by Fe(II) in absence of acid-stable cytochrome c is very slow, indicating the importance of cytochrome c in electron transport. Thus, in the iron oxidation process, acid-stable cytochrome c is the primary acceptor of electron, transferring the electron to rusticyanin at pH 2.0, which, in turn, affects electron transfer to iron-cytochrome c reductase around pH 5.5.

Immunological methods for the quantitation of the industrially important bioleaching microorganisms

To monitor the levels of Thiobacillus ferrooxidans in bioleaching operations, we have developed a specific and very sensitive dot-immunobinding assay. Polyclonal antisera against whole T. ferrooxidans cells was used, and the bacteria-antibody reaction was visualized by employing either 125I-labeled or peroxidase-conjugated protein A or 125I-labeled or peroxidase-conjunted goat anti-rabbit immunoglobulin G. A minimum of 10**3 cells per dot could be easily detected. Therfore, the method allows the sensitive, and specific simultaneous processing of numerous samples in a short time

Estudo de fatores que inibem o crescimento de ferrobactérias em poços / Factors that inhibit the growth of iron bacteria in wells

Testou-se a eficiência dos desinfetantes: cloro livre e combinado, permanganato de potássio e sulfato de cobre em baixas e altas concentraçöes por longos e curtos períodos de tempo. Verificou-se também, o efeito do pH e da aeraçäo em poços contaminados por ferrobactérias. O pH é um dos principais fatores reguladores do crescimento destes organismos. Observou-se maior crescimento em pH 6,5 e inibiçäo a partir de 7,5. Os desinfetantes se mostraram eficientes na inibiçäo do crescimento das ferrobactérias em concentraçöes a partir de 4mg/l nos expoerimentos de longos períodos de tempo e nas concentraçöes de 10mg/l a partir de 6 horas de contato