Analysis of bacterial community during the fermentation of pulque, a traditional Mexican alcoholic beverage, using a polyphasic approach

In this study, the characterization of the bacterial community present during the fermentation of pulque, a traditional Mexican alcoholic beverage from maguey (Agave), was determined for the first time by a polyphasic approach in which both culture and non-culture dependent methods were utilized. The work included the isolation of lactic acid bacteria (LAB), aerobic mesophiles, and 16S rDNA clone libraries from total DNA extracted from the maguey sap (aguamiel) used as substrate, after inoculation with a sample of previously produced pulque and followed by 6-h fermentation. Microbiological diversity results were correlated with fermentation process parameters such as sucrose, glucose, fructose and fermentation product concentrations. In addition, medium rheological behavior analysis and scanning electron microscopy in aguamiel and during pulque fermentation were also performed. Our results showed that both culture and non-culture dependent approaches allowed the detection of several new and previously reported species within the alpha-, gamma-Proteobacteria and Firmicutes. Bacteria diversity in aguamiel was composed by the heterofermentative Leuconostoc citreum, L. mesenteroides, L. kimchi, the gamma-Proteobacteria Erwinia rhapontici, Enterobacter spp. and Acinetobacter radioresistens. Inoculation with previously fermented pulque incorporated to the system microbiota, homofermentative lactobacilli related to Lactobacillus acidophilus, several alpha-Proteobacteria such as Zymomonas mobilis and Acetobacter malorum, other gamma-Proteobacteria and an important amount of yeasts, creating a starting metabolic diversity composed by homofermentative and heterofermentative LAB, acetic and ethanol producing microorganisms. At the end of the fermentation process, the bacterial diversity was mainly composed by the homofermentative Lactobacillus acidophilus, the heterofermentative L. mesenteroides, Lactococcus lactis subsp. lactis and the alpha-Proteobacteria A. malorum. After a 6-h fermentation, 83.27% of total sugars detected after inoculation were consumed (228.4 mM hexose equivalents) and a carbon (C) recovery of 66.18% in fermentation products was estimated. They were produced 284.4 mM C as ethanol, 71.5 mM C as acetic acid and 19 mM C as lactic acid, demonstrating the presence of homo- and heterofermentative, acetic and alcoholic metabolisms in the final product. It was also found, after hydrolysis, that the exopolysaccharide produced during the fermentation was mainly composed by fructose residues, probably inulin or levan.     

Effect of environmental factors and cell physiological state on Pulsed Electric Fields resistance and repair capacity of various strains of Escherichia coli

The aim was to determine the resistance variation of four strains of Escherichia coli to Pulsed Electric Fields (PEF), the role of the sigma factor RpoS in PEF resistance, as well as the influence of several environmental factors and the cell physiological state on the PEF resistance and repair capacity. The rpoS null mutant, E. coli BJ4L1, exhibited decreased PEF resistance as compared with its wild-type parent, BJ4. W3110 and O157:H7 were the most PEF-resistant strains: whereas 2 and more than 3 Log10 cycles of BJ4 and BJ4L1 cells, respectively, were inactivated after 50 pulses at 35 kV/cm, only 0.5 Log10 cycle of inactivation of W3110 and O157:H7 was attained. A different pattern was observed and the resistance variation among strains was largely reduced, when selective recovery media were used. At exponential growth phase, the resistance of the four strains was lower, and more than 4 Log10 cycles of inactivation of all strains tested were attained at 30 kV/cm. Previous heat and cold shock treatments scarcely influenced cell PEF resistance. PEF survival increased with the reduction in water activity of the treatment medium to 0.94: the occurrence of sublethally injured cells was negligible, and less than 1 Log10 cycle of inactivation was attained at 35 kV/cm. PEF-treated cells were sensitive to a subsequent storage at pH 4.0 or in the presence of sorbic acid, attaining a final inactivation of 4-5 Log10 cycles after 24 hour-incubation. In conclusion, the work confirms the role of rpoS in PEF resistance. E. coli strains exhibit large differences in PEF resistance. These differences were less important when cells were recovered under selective conditions. Both resistance variation among strains and occurrence of sublethal damage were noticeably influenced by the environmental factors tested.     

Plant-derived polyphenols attenuate lipopolysaccharide-induced nitric oxide and tumour necrosis factor production in murine microglia and macrophages

Lipopolysaccharides released during bacterial infections induce the expression of pro-inflammatory cytokines and lead to complications such as neuronal damage in the CNS and septic shock in the periphery. While the initial infection is treated by antibiotics, anti-inflammatory agents would be advantageous add-on medications. In order to identify such compounds, we have compared 29 commercially available polyphenol-containing plant extracts and pure compounds for their ability to prevent LPS-induced up-regulation of NO production. Among the botanical extracts, bearberry and grape seed were the most active preparations, exhibiting IC(50) values of around 20 mug/mL. Among the pure compounds, IC(50) values for apigenin, diosmetin and silybin were 15, 19 and 12 muM, in N-11 murine microglia, and 7, 16 and 25 muM, in RAW 264.7 murine macrophages, respectively. In addition, these flavonoids were also able to down-regulate LPS-induced tumour necrosis factor production. Structure-activity relationships of the flavonoids demonstrated three distinct principles: (i) flavonoid-aglycons are more potent than the corresponding glycosides, (ii) flavonoids with a 4'-OH substitution in the B-ring are more potent than those with a 3'-OH-4'-methoxy substitution, (iii) flavonoids of the flavone type (with a C2=C3 double bond) are more potent than those of the flavanone type (with a at C2-C3 single bond).     

玛湖凹陷风城组碱湖烃源岩生烃机理及资源量新认识

碱湖烃源岩的生烃特征和机理是学科研究的前缘与难点。以准噶尔盆地玛湖凹陷下二叠统风城组为例,采用烃源岩人工剖面、自然剖面、油气特征标定相结合的方法,对这套碱湖烃源岩的生烃特征和机理进行了研究,并据此重新计算了资源量。结果表明,风城组碱湖烃源岩的生烃特征突出表现为转化率高、连续生烃、多期高峰、生油窗长、油质轻、油多气少,不同于传统的湖相优质烃源岩。这种独特的生烃机理在于烃源岩的有机和无机组成2方面。风城组碱湖烃源岩的生烃母质以菌、藻类为主,特别是菌类发育,因此生烃具有油多气少、转化率高、连续生烃、油质轻的特色;风城组的矿物组成独特性在于兼具发育碱类矿物和火山矿物,二者对生烃分别起延滞和催化作用,使得烃源岩可以早期生烃、持续生烃、多期高峰、生油窗长。基于这种特殊的生烃特征和机理,重新评估了研究区的资源量,生油量较前期认识提高了25%,生气量减少了13%,这更贴近于目前的勘探实际,对确定下步勘探开发方向具有重要意义。     

Influence of photoperiod on carbon dioxide and methane emissions from two pilot-scale stabilization ponds

Greenhouse gas (GHG) emissions (CO(2), CH(4)) from pilot-scale algal and duckweed-based ponds (ABP and DBP) were measured using the static chamber methodology. Daylight and nocturnal variations of GHG and wastewater characteristics (e.g. chemical oxygen demand (COD), pH) were determined via sampling campaigns during midday (12:30-15:30) and midnight (00:30-03:30) periods. The results showed that under daylight conditions in ABP median emissions were -232 mg CO(2) m(-2) d(-1) and 9.9 mg CH(4) m(-2) d(-1), and in DBP median emissions were -1,654.5 mg CO(2) m(-2) d(-1) and 71.4 mg CH(4) m(-2) d(-1), respectively. During nocturnal conditions ABP median emissions were 3,949.9 mg CO(2) m(-2) d(-1), 12.7 mg CH(4) m(-2) d(-1), and DBP median emissions were 5,116 mg CO(2) m(-2) d(-1), 195.2 mg CH(4) m(-2) d(-1), respectively. Once data measured during daylight were averaged together with nocturnal data the median emissions for ABP were 1,566.8 mg CO(2) m(-2) d(-1) and 72.1 mg CH(4) m(-2) d(-1), whilst for DBP they were 3,016.9 mg CO(2) m(-2) d(-) and 178.9 mg CH(4) m(-2) d(-1), respectively. These figures suggest that there were significant differences between CO(2) emissions measured during daylight and nocturnal periods (p < 0.05). This shows a sink-like behaviour for both ABP and DBP in the presence of solar light, which indicates the influence of photosynthesis in CO(2) emissions. On the other hand, the fluxes of CH(4) indicated that DBP and ABP behave as net sources of CH(4) during day and night, although higher emissions were observed from DBP. Overall, according to the compound average (daylight and nocturnal emissions) both ABP and DBP systems might be considered as net sources of GHG.     

n-Alkane content of intramuscular lipids of Iberian fresh ham from different feeding systems and crossbreeding

n-Alkane content of intramuscular lipids (Biceps femoris muscle) of the Iberian pig have been determined. Thirty-four pigs, divided into four groups, based in the feeding system (Montanera, fed on acorns and pasture; and Pienso, fed on a concentrate feed) and in the genotype (Iberian pure pigs; and Iberian crossbred with Duroc 50%) were studied. n-Alkane content of intramuscular lipids has not been affected by neither crossbreeding nor feeding, although the analysis of feeds administered to the pigs showed greater n-alkane values in pasture (consumed by animals in montanera), than in acorns and concentrate feed.     

Isolation and sequence of the MIG1 homologue from the yeast Candida utilis

The Mig1p repressor from the food yeast Candida utilis has been isolated using a homologous PCR hybridization probe. This probe was amplified with two sets of degenerate primers designed on the basis of highly conserved motifs in the DNA-binding region (zinc-finger domain) from yeast Mig1p and fungi CreA repressors. The cloned gene was sequenced and found to encode a polypeptide of 345 amino acids which shows significant identity with other yeast and fungus repressors in the DNA-binding domain and also with the yeast Mig1 proteins in the C-terminal region (effector domain). The MIG1 repressor gene from C. utilis was able to complement functionally the mig1 mutation of S. cerevisiae. The sequence presented here has been deposited in the EMBL data library under Accession No. AJ277830.     

Contribution of starter culture to the sensory characteristics of fermented Almagro eggplants

Trial fermentations were performed using three experimental starter cultures with a view to selecting the most appropriate starter for use in the manufacture of Almagro eggplants. The lactic acid bacterial strains used in the starters had previously been isolated from spontaneous fermentations. The combined action of the obligate heterofermentative species Lactobacillus brevis and the facultative heterofermentative species L. plantarum yielded Almagro eggplants with sensory properties preferred by panelists. Conversely, another obligate heterofermentative species, namely, L. fermentum, present in certain starter formulations tested, appeared to encounter difficulty growing during fermentation and exerted little influence on the sensory characteristics of the eggplants produced.     

Mechanical resistance properties of gravel used in subsurface flow constructed wetlands: implications for clogging

Gravel constitutes the filter medium in subsurface flow constructed wetlands (SSF CWs) and its porosity and hydraulic conductivity decrease over time (clogging), limiting the lifespan of the systems. Using gravel of poor quality accelerates clogging in wetlands. In this study, gravel samples from six different wetland systems were compared with regards to their mineral composition and mechanical resistance properties. Results showed that both mineralogy and texture are related to mechanical resistance. Accordingly, gravel with high content of quartz (> 80%) showed a lower percentage of broken particles (0.18-1.03%) than those with lower content of quartz (2.42-4.56% media broken). Although granite is formed by high durability minerals, its non-uniform texture results in a lower resistance to abrasion (ca. 10% less resistance than calcareous gravel). Therefore, it is recommended to use gravels composed mainly of quartz or, when it is not available, limestone gravels (rounded and uniform) are recommended instead. The resistance to abrasion (LAA test) seems to be a good indicator to determine the mechanical properties of gravels used in CWs. It is recommended to use gravels with LAA below 30% in order to avoid a rapid clogging due to gravel crumbling and subsequent mineral solids accumulation.