José Luis Sanz

Phenotypic Properties and Microbial Diversity of Methanogenic Granules from a Full-Scale Upflow Anaerobic Sludge Bed Reactor Treating Brewery Wastewater

ABSTRACT Methanogenic granules from an anaerobic bioreactor that treated wastewater of a beer brewery consisted of different morphological types of granules. In this study, the microbial compositions of the different granules were analyzed by molecular microbiological techniques: cloning, denaturing gradient gel electrophoresis and fluorescent in situ hybridization (FISH), and scanning and transmission electron microscopy. We propose here that the different types of granules reflect the different stages in the life cycle of granules. Young granules were small, black, and compact and harbored active cells. Gray granules were the most abundant granules. These granules have a multilayer structure with channels and void areas. The core was composed of dead or starving cells with low activity. The brown granules, which were the largest granules, showed a loose and amorphous structure with big channels that resulted in fractured zones and corresponded to the older granules. Firmicutes (as determined by FISH) and Nitrospira and Deferribacteres (as determined by cloning and sequencing) were the predominant Bacteria. Remarkably, Firmicutes could not be detected in the brown granules. The methanogenic Archaea identified were Methanosaeta concilii (70 to 90% by FISH and cloning), Methanosarcina mazei, and Methanospirillum spp. The phenotypic appearance of the granules reflected the physiological condition of the granules. This may be valuable to easily select appropriate seed sludges to start up other reactors.

Sulfate reduction at low pH to remediate acid mine drainage.

Industrial activities and the natural oxidation of metallic sulfide-ores produce sulfate-rich waters with low pH and high heavy metals content, generally termed acid mine drainage (AMD). This is of great environmental concern as some heavy metals are highly toxic. Within a number of possibilities, biological treatment applying sulfate-reducing bacteria (SRB) is an attractive option to treat AMD and to recover metals. The process produces alkalinity, neutralizing the AMD simultaneously. The sulfide that is produced reacts with the metal in solution and precipitates them as metal sulfides. Here, important factors for biotechnological application of SRB such as the inocula, the pH of the process, the substrates and the reactor design are discussed. Microbial communities of sulfidogenic reactors treating AMD which comprise fermentative-, acetogenic- and SRB as well as methanogenic archaea are reviewed.

Effect of simultaneous use of highly active antiretroviral therapy on survival of HIV patients with tuberculosis.

Introduction:The optimal timing for initiation of highly active antiretroviral therapy (HAART) in patients with AIDS and tuberculosis (TB) is an unresolved question. To assess the effect of HAART on the survival of patients with TB, we designed this study. Methods:We selected all HIV patients included in the COMESEM cohort with TB diagnosis after 1996. Clinical and epidemiological data were registered. We compared patients who started HAART at the diagnosis of TB [simultaneous therapy (ST)] or not. Survival was assessed by Cox analysis. Results:Among the 6934 HIV patients included in the cohort, 1217 patients had TB, 322 of them (26.5%) after 1996. At the time of TB diagnosis, 45% of them started HAART (ST). There were no differences between groups regarding basal characteristics, except for a lower viral load in ST patients. ST therapy was associated with improved survival (hazard ratio 0.38; 95% confidence interval 0.20 to 0.72, P = 0.003). By univariate analysis, survival was also associated with no endovenous drug use and a later year of TB diagnosis. After adjusting for other prognostic variables, by Cox multivariate analysis, ST remained robustly associated with improved survival (hazard ratio 0.37; 95% confidence interval 0.17 to 0.66, P = 0.001). Conclusions:Simultaneous HAART and TB treatment in HIV patients with TB is associated with improved survival.

Microbial Diversity in Anaerobic Sediments at Río Tinto, a Naturally Acidic Environment with a High Heavy Metal Content

ABSTRACT The Tinto River is an extreme environment located at the core of the Iberian Pyritic Belt (IPB). It is an unusual ecosystem due to its size (100 km long), constant acidic pH (mean pH, 2.3), and high concentration of heavy metals, iron, and sulfate in its waters, characteristics that make the Tinto River Basin comparable to acidic mine drainage (AMD) systems. In this paper we present an extensive survey of the Tinto River sediment microbiota using two culture-independent approaches: denaturing gradient gel electrophoresis and cloning of 16S rRNA genes. The taxonomic affiliation of the Bacteria showed a high degree of biodiversity, falling into 5 different phyla: Proteobacteria, Firmicutes, Bacteroidetes, Acidobacteria, and Actinobacteria; meanwhile, all the Archaea were affiliated with the order Thermoplasmatales. Microorganisms involved in the iron (Acidithiobacillus ferrooxidans, Sulfobacillus spp., Ferroplasma spp., etc.), sulfur (Desulfurella spp., Desulfosporosinus spp., Thermodesulfobium spp., etc.), and carbon (Acidiphilium spp., Bacillus spp., Clostridium spp., Acidobacterium spp., etc.) cycles were identified, and their distribution was correlated with physicochemical parameters of the sediments. Ferric iron was the main electron acceptor for the oxidation of organic matter in the most acid and oxidizing layers, so acidophilic facultative Fe(III)-reducing bacteria appeared widely in the clone libraries. With increasing pH, the solubility of iron decreases and sulfate-reducing bacteria become dominant, with the ecological role of methanogens being insignificant. Considering the identified microorganisms—which, according to the rarefaction curves and Goods coverage values, cover almost all of the diversity—and their corresponding metabolism, we suggest a model of the iron, sulfur, and organic matter cycles in AMD-related sediments.

A simplification trial switching from nucleoside reverse transcriptase inhibitors to once-daily fixed-dose abacavir/lamivudine or tenofovir/emtricitabine in HIV-1-infected patients with virological suppression.

Background:Data comparing abacavir/lamivudine versus tenofovir/emtricitabine in antiretroviral-naive patients are controversial. We compared 48-week efficacy and safety of these combinations as substitutes of nucleosides in patients with virological suppression. Methods:We randomly assigned 333 HIV-1-infected patients on lamivudine-containing triple regimens with <200 copies per milliliter for at least 6 months to switch their nucleosides to either abacavir/lamivudine (n = 167) or tenofovir/emtricitabine (n = 166). The primary outcome was treatment failure [“switching = failure” intention to treat (ITT) analysis, noninferiority margin 12.5%]. Secondary outcomes were time to treatment failure, virological failure, adverse events, and changes in CD4 count, fasting plasma lipids, lipodystrophy, body fat, bone mineral density, and renal function. Results:Treatment failure occurred in 32 patients (19%) on abacavir/lamivudine and 22 patients (13%) on tenofovir/emtricitabine [difference 5.9%; (95% confidence interval −2.1% to 14.0%), P = 0.06]. Four patients in the abacavir/lamivudine group versus none in the tenofovir/emtricitabine group developed virological failure [difference 2.4; (95% confidence interval 0.05 to 6.0), P = 0.04]. Twenty-three patients (14%) assigned to abacavir/lamivudine and 10 (6%) to tenofovir/lamivudine experienced grade 3 or 4 adverse effects (P = 0.03). CD4 counts and plasma lipids showed higher increments in the abacavir/lamivudine group than in the tenofovir/emtricitabine group. Conclusions:In HIV-1-infected patients with virological suppression, abacavir/lamivudine did not meet the noninferiority outcome for treatment efficacy compared with tenofovir/emtricitabine.

The action of antibiotics on the anaerobic digestion process.

Abstract Antibiotics can disturb the production of biogas during anaerobic digestion. This study shows a systematic approach to understanding how the different bacterial populations involved in the final conversion of organic matter into methane are inhibited by 15 antimicrobial agents with different specificities and modes of action. The results obtained show the following trends: (i) some inhibitors, such as the macrolide erythromycin, lack any inhibitory effect on biogas production; (ii) some antibiotics, with different specificities, have partial inhibitory effects on anaerobic digestion and decrease methane production by interfering with the activity of propionic-acid- and butyric-acid-degrading bacteria,␣(e.g. antibiotics that interfere with cell wall synthesis, RNA polymerase activity and protein synthesis, especially the aminoglycosides); (iii) the protein synthesis inhibitors chlortetracycline (IC50 40 mg l−1) and chloramphenicol (IC50 15–20 mg l−1) are very powerful inhibitors of anaerobic digestion. The majority of the antibiotics tested lacked activity against acetoclastic methanogens, being active only on the acetogenic bacteria. However, chloramphenicol and chlortetracycline could cause the complete inhibition of the acetoclastic methanogenic archaea.

An Early Cretaceous faunal and floral continental assemblage: Las Hoyas fossil site (Cuenca, Spain)

Abstract The fossiliferous beds of Las Hoyas (province ofCuenca, Spain) are composed of limestones (probably Late Hauterivian). The main fossiliferous facies consists of laminated limestone from a lake-basin plain with bottom waters perennially anoxic. The floral and faunal assemblage is similar to that of Montsech (prov. of Lerida, Spain), with divergences especially in the insects and some tetrapods. Bottom-dwelling invertebrates are scarce. Most of invertebrates are arthropods: three crustaceans and a relatively diversified entomofauna. The fishes are the main component of the vertebrate fauna both in number of individuals and diversity (13 different taxa). Among tetrapods the most abundant is a new genus of Caudata. Reptiles are represented by a chelonian, a little lizard and an atoposaurid crocodile. Finally, a new bird is reported from Las Hoyas, with an intermediate phylogenetic position between Archaeopteryx and Ornithurae.

On the presence of Allosaurus fragilis (Theropoda: Carnosauria) in the Upper Jurassic of Portugal: first evidence of an intercontinental dinosaur species

The species Allosaurus fragilis, from the Morrison Formation of North America (Kimmeridgian–Tithonian, Upper Jurassic), is one of the best known members of the Theropoda, the group including all predatory non-avian dinosaurs and birds. Here, we report on the first diagnostic theropod remains discovered from the Jurassic of the Iberian Peninsula. The specimen is the first evidence of A. fragilis outside North America. Thus, this taxon represents the first dinosaur species found on two different continents, and suggests the existence of a ‘land bridge’ between North America and Europe during the Late Jurassic.

Comparison of UASB and EGSB performance on the anaerobic biodegradation of 2,4-dichlorophenol

The anaerobic degradation of 2,4-dichlorophenol (2,4-DCP) in upflow anaerobic sludge blanket (UASB) and expanded granular sludge bed (EGSB) reactors using glucose as main carbon source was studied. The performance of both systems was compared in terms of 2,4-DCP and COD removal efficiencies, methane production, stability, granular sludge adaptability as well as reversion of the bacterial inhibition. Both organic and 2,4-DCP loading rates were incrementally varied through the experiments. With loading rates of 1.9 gCODL(-1)d(-1) and 100mg 2,4-DCP L(-1)d(-1), 75% and 84% removal efficiencies of this compound, accompanied by COD consumption efficiencies of 61% and 80% were achieved in the UASB and EGSB reactors, respectively. In these conditions, methane production reached 0.088 L CH(4)g(-1) COD in the EGSB reactor whereas in the UASB reactor was almost negligible. Decreasing the 2,4-DCP loading rate to 30 mgL(-1)d(-1) an improvement in the methane production was observed in both reactors (methanogenic activity of 0.148 and 0.192 L CH(4)g(-1) COD in UASB and EGSB reactors, respectively). Efficiency of dechlorination was improved in both reactors from around 30% to 80% by reducing to one-half the COD due to a decreasing of the 4-chlorophenol concentration accumulated in the effluents of both reactors. The dechlorination efficiency of the UASB reactor was dramatically inhibited at a 2,4-DCP feed concentration above around 210 mgL(-1) because of 2,4-DCP accumulation in the effluent. SEM studies revealed no significant morphological changes in the sludge granules.

Prokaryotic diversity and dynamics in a full-scale municipal solid waste anaerobic reactor from start-up to steady-state conditions.

The prokaryotic diversity of an anaerobic reactor for the treatment of municipal solid waste was investigated over the course of 2 years with the use of 16S rDNA-targeted molecular approaches. The fermentative Bacteroidetes and Firmicutes predominated, and Proteobacteria, Actinobacteria, Tenericutes and the candidate division WWE1 were also identified. Methane production was dominated by the hydrogenotrophic Methanomicrobiales (Methanoculleus sp.) and their syntrophic association with acetate-utilizing and propionate-oxidizing bacteria. qPCR demonstrated the predominance of the hydrogenotrophic over aceticlastic Methanosarcinaceae (Methanosarcina sp. and Methanimicrococcus sp.), and Methanosaetaceae (Methanosaeta sp.) were measured in low numbers in the reactor. According to the FISH and CARD-FISH analyses, Bacteria and Archaea accounted for 85% and 15% of the cells, respectively. Different cell counts for these domains were obtained by qPCR versus FISH analyses. The use of several molecular tools increases our knowledge of the prokaryotic community dynamics from start-up to steady-state conditions in a full-scale MSW reactor.