PCR-based DGGE and FISH analysis of methanogens in an anaerobic closed digester tank for treating palm oil mill effluent

16S ribosomal RNA (rRNA)-targeted fluorescent in situ hybridization combined with polymerase chain reaction (PCR)-cloning, light microscopy using Gram stains, scanning electron microscopy and denatured gradient gel electrophoresis were used to reveal the distribution of methanogens within an anaerobic closed digester tank fed with palm oil mill effluent. For specific detection of methanogens, 16S rRNA-cloning analysis was conducted followed by restriction fragment length polymorphism (RFLP) for presumptive identification of methanogens. To cover the drawbacks of the PCR-cloning study, the organization of the microorganisms was visualized in the activated sludge sample by using fluorescent oligonucleotide probes specific to several different methanogens, and a probe for bacteria. In situ hybridization with methanogens and bacterial probes and denatured gradient gel electrophoresis within activated sludge clearly confirmed the presence of Methanosaeta sp. and Methanosarcina sp. cells. Methanosaeta concilii was found to be the dominant species in the bioreactor. These results revealed the presence of possibly new strain of Methanosaeta in the bioreactor for treating palm oil mill effluent called Methanosaeta concilii SamaliEB (Gene bank accession number: EU580025). In addition, fluorescent hybridization pictured the close association between the methanogens and bacteria and that the number of methanogens was greater than the number of bacteria.

Performance of an Anaerobic Baffled Reactor (ABR) in treatment of cassava wastewater / Desempenho de um Reator Anaeróbio Compartimentado (RAC) no tratamento da manipueira

The performance of an anaerobic baffled reactor (ABR) was evaluated in the treatment of cassava wastewater, a pollutant residue. An ABR divided in four equal volume compartments (total volume 4L) and operated at 35ºC was used in cassava wastewater treatment. Feed tank chemical oxygen demand (COD) was varied from 2000 to 7000 mg L-1 and it was evaluated the most appropriated hydraulic retention time (HRT) for the best performance on COD removal. The ABR was evaluated by analysis of COD (colorimetric method), pH, turbidity, total and volatile solids, alkalinity and acidity. Principal component analysis (PCA) was carried to better understand data obtained. The system showed buffering ability as acidity decreased along compartments while alkalinity and pH values were increased. There was particulate material retention and COD removal varied from 83 to 92% for HRT of 3.5 days.

O desempenho de um reator anaeróbio compartimentado (RAC) foi avaliado para o tratamento da manipueira, resíduo tóxico. Um RAC dividido em quatro compartimentos de mesmo volume (volume total 4L) e operado a 35ºC foi utilizado no tratamento da manipueira. A demanda química de oxigênio (DQO) do tanque de alimentação variou de 2000 a 7000 mg L-1 e avaliou-se o tempo de residência hidráulica (TRH) mais apropriado ao melhor desempenho do reator. O RAC foi avaliado pelas análises de DQO (método colorimétrico), pH, turbidez, sólidos totais e voláteis, alcalinidade e acidez. A Análise das Componentes Principais (PCA) foi conduzida para melhor compreensão dos dados obtidos. O sistema apresentou capacidade tamponante conforme a acidez decresceu ao longo dos compartimentos ao passo que a alcalinidade e o pH aumentaram. Houve retenção do material particulado e a remoção da DQO variou de 83 a 92% para TRH de 3,5 dias.

Macromolecular composition and anaerobic degradation of the sludge produced in a sequencing batch reactor

The effect of sequencing batch reactor (SBR) operating conditions on sludge macromolecular composition and the effect of sludge macromolecular composition on the anaerobic degradation of the sludge produced in SBR was investigated in this work. A SBR, fed with synthetic wastewater, was operated at different air flow rates. The resulting sludge was analyzed in terms of protein, carbohydrate, phospholipid and polyhydroxybutyrate concentrations. Methane production during anaerobic digestion of the sludge was also measured. Ammonium, nitrite, nitrate, dissolved oxygen and chemical oxygen demand (COD) track studies in the SBR were carried out in order to relate SBR performance and sludge macromolecular composition. The lowest air flow rate at which the SBR was operated was 2 l min-1, in which case the dissolved oxygen concentration was lower than 0.5 mg l-1 in the SBR and partial denitrification occurred during the feeding phase. An increased air flow rate caused a decrease in protein concentration, as well as an increase in carbohydrate concentration. Polyhydroxybutyrate (PHB) concentration in the sludge was independent of air flow rate. At different air flows, the methane production rates were similar, but the total volume of methane was greater during anaerobic digestion of the sludge produced at low air flow rates. These results indicate a strategy by which changes in sludge composition can optimize the operation of anaerobic sludge digesters.

Descomposición de hojas de Tessaria integrifolia (Asteraceae) y colonización por invertebrados en un cause secundario del río Paraná medio / Descomposition and invertebrate colonization of Tessaria integrifolia (Asteraceae) leaves in a secondary channel of the middle Paraná river

Se analizó la descomposición de hojas de Tessaria integrifolia y su colonización por invertebrados en el río Tiradero Viejo, cauce secundario del río Paraná, Argentina. Las hojas de T. integrifolia fueron recolectadas después de la abscisión; se colocaron 6g de hojas secadas al aire en bolsas de nylon de 5mm de abertura de malla y fueron expuestas al proceso de descomposición en la ribera del río. Se extrajeron 3 réplicas a los 7, 14, 28, 56 y 112 días. El coeficiente de descomposición (k) obtenido fue de 0,0143 a los 112 días. La densidad máxima de invertebrados fue de 395 individuos/g de peso seco de hojas a los 56 días de exposición. La mayor riqueza de especies y diversidad fueron obtenidas a los 28 días, siendo los oligoquetos (Pristina leidyi, P. aequiseta, Dero multibranchiata), quironómidos (Polypedilum spp., Phaenopsectra sp.) y bivalvos (Limnoperna fortunei) los taxa dominantes. Los colectores-recolectores fueron dominantes durante las primeras etapas de la sucesión y los colectores-filtradores aumentaron en las etapas finales, mientras que los trituradores fueron escasos pero más constantes a lo largo de la experiencia.