Out of the blue: the independent activity of sulfur-oxidizers and diatoms mediate the sudden color shift of a tropical river.

BACKGROUND: Río Celeste ("Sky-Blue River") is a river located in the Tenorio National Park (Costa Rica) that has become an important hotspot for eco-tourism due to its striking sky-blue color. A previous study indicated that this color is not caused by dissolved chemical species, but by formation of light-scattering aluminosilicate particles at the mixing point of two colorless streams, the acidic Quebrada Agria and the neutral Río Buenavista. RESULTS: We now present microbiological information on Río Celeste and its two tributaries, as well as a more detailed characterization of the particles that occur at the mixing point. Our results overturn the previous belief that the light scattering particles are formed by the aggregation of smaller particles coming from Río Buenavista, and rather point to chemical formation of hydroxyaluminosilicate colloids when Quebrada Agria is partially neutralized by Río Buenavista, which also contributes silica to the reaction. The process is mediated by the activities of different microorganisms in both streams. In Quebrada Agria, sulfur-oxidizing bacteria generate an acidic environment, which in turn cause dissolution and mobilization of aluminum and other metals. In Río Buenavista, the growth of diatoms transforms dissolved silicon into colloidal biogenic forms which may facilitate particle precipitation. CONCLUSIONS: We show how the sky-blue color of Río Celeste arises from the tight interaction between chemical and biological processes, in what constitutes a textbook example of emergent behavior in environmental microbiology.

Temperature and elemental sulfur shape microbial communities in two extremely acidic aquatic volcanic environments.

Aquatic environments of volcanic origin provide an exceptional opportunity to study the adaptations of microorganisms to early planet life conditions. Here, we characterized the prokaryotic communities and physicochemical properties of seepage sites at the bottom of the Poas Volcano crater and the Agrio River, two geologically related extremely acidic environments located in Costa Rica. Both locations hold a low pH (1.79-2.20) and have high sulfate and iron concentrations (Fe = 47-206 mg/L, SO42- = 1170-2460 mg/L), but significant differences in their temperature (90.0-95.0 ºC in the seepages at Poas Volcano, 19.1-26.6 ºC in Agrio River) and in the elemental sulfur content. Based on the analysis of 16S rRNA gene sequences, we determined that Sulfobacillus spp. represented more than half of the sequences in Poas Volcano seepage sites, while Agrio River was dominated by Leptospirillum and members of the archaeal order Thermoplasmatales. Both environments share some chemical characteristics and part of their microbiota, however, the temperature and the reduced sulfur are likely the main distinguishing features, ultimately shaping their microbial communities. Our data suggest that in the Poas Volcano-Agrio River system there is a common metabolism but with specialization of species that adapt to the physicochemical conditions of each environment.

Microbial Community Structure Along a Horizontal Oxygen Gradient in a Costa Rican Volcanic Influenced Acid Rock Drainage System.

We describe the geochemistry and microbial diversity of a pristine environment that resembles an acid rock drainage (ARD) but it is actually the result of hydrothermal and volcanic influences. We designate this environment, and other comparable sites, as volcanic influenced acid rock drainage (VARD) systems. The metal content and sulfuric acid in this ecosystem stem from the volcanic milieu and not from the product of pyrite oxidation. Based on the analysis of 16S rRNA gene amplicons, we report the microbial community structure in the pristine San Cayetano Costa Rican VARD environment (pH = 2.94-3.06, sulfate ~ 0.87-1.19 g L-1, iron ~ 35-61 mg L-1 (waters), and ~ 8-293 g kg-1 (sediments)). San Cayetano was found to be dominated by microorganisms involved in the geochemical cycling of iron, sulfur, and nitrogen; however, the identity and abundance of the species changed with the oxygen content (0.40-6.06 mg L-1) along the river course. The hypoxic source of San Cayetano is dominated by a putative anaerobic sulfate-reducing Deltaproteobacterium. Sulfur-oxidizing bacteria such as Acidithiobacillus or Sulfobacillus are found in smaller proportions with respect to typical ARD. In the oxic downstream, we identified aerobic iron-oxidizers (Leptospirillum, Acidithrix, Ferrovum) and heterotrophic bacteria (Burkholderiaceae bacterium, Trichococcus, Acidocella). Thermoplasmatales archaea closely related to environmental phylotypes found in other ARD niches were also observed throughout the entire ecosystem. Overall, our study shows the differences and similarities in the diversity and distribution of the microbial communities between an ARD and a VARD system at the source and along the oxygen gradient that establishes on the course of the river.

Photonic Crystal Characterization of the Cuticles of Chrysina chrysargyrea and Chrysina optima Jewel Scarab Beetles.

A unified description involving structural morphology and composition, dispersion of optical constants, modeled and measured reflection spectra and photonic crystal characterization is devised. Light reflection spectra by the cuticles of scarab beetles (Chrysina chrysargyrea and Chrysina optima), measured in the wavelength range 300-1000 nm, show spectrally structured broad bands. Scanning electron microscopy analysis shows that the pitches of the twisted structures responsible for the left-handed circularly polarized reflected light change monotonically with depth through the cuticles, making it possible to obtain the explicit depth-dependence for each cuticle arrangement considered. This variation is a key aspect, and it will be introduced in the context of Berreman's formalism, which allows us to evaluate reflection spectra whose main features coincide in those displayed in measurements. Through the dispersion relation obtained from the Helmholtz's equation satisfied by the circular components of the propagating fields, the presence of a photonic band gap is established for each case considered. These band gaps depend on depth through the cuticle, and their spectral positions change with depth. This explains the presence of broad bands in the reflection spectra, and their spectral features correlate with details in the variation of the pitch with depth. The twisted structures consist of chitin nanofibrils whose optical anisotropy is not large enough so as to be approached from modeling the measured reflection spectra. The presence of a high birefringence substance embedded in the chitin matrix is required. In this sense, the presence of uric acid crystallites through the cuticle is strongly suggested by frustrated attenuated total reflection and Raman spectroscopy analysis. The complete optical modeling is performed incorporating the wavelength-dependent optical constants of chitin and uric acid.

Pristine but metal-rich Río Sucio (Dirty River) is dominated by Gallionella and other iron-sulfur oxidizing microbes.

Whether the extreme conditions of acidity and heavy metal pollution of streams and rivers originating in pyritic formations are caused primarily by mining activities or by natural activities of metal-oxidizing microbes living within the geological formations is a subject of considerable controversy. Most microbiological studies of such waters have so far focused on acid mine drainage sites, which are heavily human-impacted environments, so it has been problematic to eliminate the human factor in the question of the origin of the key metal compounds. We have studied the physico-chemistry and microbiology of the Río Sucio in the Braulio Carrillo National Park of Costa Rica, 22 km from its volcanic rock origin. Neither the remote origin, nor the length of the river to the sampling site, have experienced human activity and are thus pristine. The river water had a characteristic brownish-yellow color due to high iron-dominated minerals, was slightly acidic, and rich in chemolithoautotrophic iron- and sulfur-oxidizing bacteria, dominated by Gallionella spp. Río Sucio is thus a natural acid-rock drainage system whose metal-containing components are derived primarily from microbial activities.

Characterization of NO adducts of the diiron center in protein R2 of Escherichia coli ribonucleotide reductase and site-directed variants; implications for the O2 activation mechanism.

The R2 subunit of Escherichia coli ribonucleotide reductase contains a diiron site that reacts with O(2) to produce a tyrosine radical (Y122.). In wild-type R2 (R2-wt), the first observable reaction intermediate is a high-valent [Fe(III)-Fe(IV)] state called compound X, but in related diiron proteins such as methane monooxygenase, Delta(9)-desaturase, and ferritin, peroxodiiron(III) complexes have been characterized. Substitution of iron ligand D84 by E within the active site of R2 allows an intermediate (mu-1,2-peroxo)diiron species to accumulate. To investigate the possible involvement of a bridging peroxo species within the O(2) activation sequence of R2-wt, we have characterized the iron-nitrosyl species that form at the diiron sites in R2-wt, R2-D84E, and R2-W48F/D84E by using vibrational spectroscopy. Previous work has shown that the diiron center in R2-wt binds one NO per iron to form an antiferromagnetically coupled [(FeNO)(7)](2) center. In the wt and variant proteins, we also observe that both irons bind one NO to form a (FeNO)(7) dimer where both Fe-N-O units share a common vibrational signature. In the wt protein, nu(Fe-NO), delta(Fe-N-O), and nu(N-O) bands are observed at 445, 434 and 1742 cm(-1), respectively, while in the variant proteins the nu(Fe-NO) and delta(Fe-N-O) bands are observed approximately 10 cm(-1) higher and the nu(N-O) approximately 10 cm(-1) lower at 1735 cm(-1). These results demonstrate that all three proteins accommodate fully symmetric [(FeNO)(7)](2) species with two identical Fe-N-O units. The formation of equivalent NO adducts in the wt and variant proteins strongly favors the formation of a symmetric bridging peroxo intermediate during the O(2) activation process in R2-wt.

Complexes of the Trioxodinitrate Anion: Synthesis and Characterization of [Zn(II)(bipy)(H(2)O)(N(2)O(3))] and [Co(II)(bipy)(2)(N(2)O(3))].

The first two complexes of the trioxodinitrate anion were prepared by reaction of Zn(bipy)Cl(2).1.5H(2)O (1.1.5H(2)O) with Na(2)N(2)O(3).H(2)O in water to produce Zn(II)(bipy)(H(2)O)(N(2)O(3)) (2) and an analogous reaction of Co(bipy)(2)Cl(2).2.5H(2)O (3.2.5H(2)O) to produce Co(II)(bipy)(2)(N(2)O(3)) (4). Both complexes are air-sensitive and crystallize in the monoclinic space group P2(1)/n. For 2.H(2)O T = 24-26 degrees C, Z = 4, a = 7.7595(6) Å, b = 9.4798(7) Å, c = 17.197(1) Å, beta = 98.142(7) degrees, and V = 1252.3(2) Å(3). The final residuals were R1 = 0.0452, wR2 = 0.1172, and quality-of-fit = 1.057. The final model consisted of the main structure and a minor, 4% disordered component. For 4.4H(2)O T = -123 degrees C, Z = 4, a = 9.0950(9) Å, b = 17.924(3) Å, c = 13.695(2) Å, beta = 90.32(1) degrees, and V = 2232.5(6) Å(3). The final least-squares residuals were R1 = 0.0859, wR2 = 0.1975, and quality-of-fit = 0.917. A network of hydrogen bonds involving the crystal water molecules and the coordinated trioxodinitrate anion was found in each case; in 4, a series of fused four- and eight-membered rings were found to be a dominant feature in the extended structure. The structural data and the IR spectrum indicate that the N-N bond of trioxodinitrate, 1.264(5) Å in the free anion, is longer for the coordinated anion (1.280(5) Å in 2 and 1.333(11) Å in 4).