Influence of nutrient, toxic metal and herbicide contents on the soil bacterial communities in tropical vegetable growing areas.

The relationship between bacterial diversity and the bioavailability of nutrients, toxic metals and the herbicide oxyfluorfen in a tropical vegetable growing area was evaluated. The study was conducted in a vegetable growing area located in the mountainous region of Rio de Janeiro (Brazil), and samples were collected in areas of vegetable cultivation and areas of environmental reserve. Fertility analyses and determination of the pseudototal levels of toxic metals in the soil samples were performed. The profile of the soil bacterial community was determined by amplification of the 16S rRNA gene and separation by DGGE. The results showed that the levels of toxic metals and elements associated with soil fertility were higher in vegetable production areas. These differences in the physical and chemical characteristics of the soil favored the presence of a greater number of OTUs in the cultivation areas (17.3-27 OTUs) than in the areas of environmental reserve (13-22 OTUs). Therefore, this study demonstrates that the presence of toxic metals and the herbicide oxyfluorfen and the increase in fertility in soils in areas with intensive vegetable cultivation resulting from the intensive management adopted in these areas promotes a differentiation of the bacterial profiles in soils in tropical vegetable growing areas.

Hyperthermophile diversity microbes in the Calientes geothermal field, Tacna, Peru.

Hyperthermophile microorganisms have been discovered worldwide, and several studies regarding biodiversity and the potential biotechnological applications have been reported. In this work, we describe for the first time the diversity of hyperthermophile communities in the Calientes Geothermal Field (CGF) located 4400 m above sea level in Tacna Region, Perú. Three hot springs were monitored and showed a temperature around 84 to 88 °C, for the microbiome analyzed was taken by sampling of sediment and water (pH 7.3-7.6). The hyperthermophile diversity was determined by PCR, DGGE, and DNA sequencing. The sediments analyzed showed a greater diversity than water samples. Sediments showed a more abundant population of bacteria than archaea, with the presence of at least 9 and 5 phylotypes, respectively. Most interestingly, in some taxa of bacteria (Bacillus) and archaea (Haloarcula and Halalkalicoccus), any of operational taxonomic units (OTUs) have not been observed before in hyperthermophile environments. Our results provide insight in the hyperthermophile diversity and reveal the possibility to develop new biotechnological applications based on the kind of environments.

Microbial Community Profiling Using Terminal Restriction Fragment Length Polymorphism (T-RFLP) and Denaturing Gradient Gel Electrophoresis (DGGE).

In their natural environments, microorganisms usually live in organized communities. Profiling analysis of microbial communities has recently assumed special relevance as it allows a thorough understanding of the diversity of the microbiota, its behavior over time, and the establishment of patterns associated with health and disease. The application of molecular biology approaches holds the advantage of including culture-difficult and as-yet-uncultivated phylotypes in the profiles, providing a more comprehensive picture of the microbial community. This chapter focuses on two particular techniques, namely terminal restriction fragment length polymorphism (T-RFLP) and denaturing gradient gel electrophoresis (DGGE), both of which have been widely used in environmental studies and have been recently successfully used by the authors in the study of the oral microbial communities associated with conditions of health and disease.

Bacterial diversity and detection of resistance genes to broad-spectrum betalactams in dairy family farm soils / Diversidade bacteriana e detecção de genes de resistência a betalactâmicos de amplo espectro em solos de propriedade familiar leiteira

Bovine mastitis is a complex disease that brings great losses to the dairy producer. The microbial diversity of the soils, as well as the presence of resistance genes in the environment directly influence the maintenance of mastitis in the farm. The objective of this work was to analyze the bacterial diversity in pasture soils of a dairy family farm, detecting enterobacteria that may be involved in the etiology of bovine mastitis, and to detect genes that encode broad-spectrum betalactamases in these soils. Twelve soil samples, representative of different areas of the farm located in the municipality of Barra do Piraí, Rio de Janeiro, were collected at different times of the year. Total DNA was extracted from the samples, gene amplified by Nested-PCR and then the amplification products were separated by DGGE (Denaturing Gradient Gel Electrophoresis). With the DGGE it was possible to construct dendograms that effectively represented the bacterial diversity of these soils. Eight of the soil samples were used to amplify the genes encoding the betalactamase enzymes TEM (blaTEM gene), SHV (blaSHV gene) and CTX (blaCTXM gene). In three of the eight soil samples, the blaSHV gene was found to be present. The blaTEM and blaCTX-M genes were not detected in any of the samples. The detection of genes encoding broad-spectrum betalactamases in dairy cattle pasture soils is of concern, because the transfer of gene material between pathogenic and non-pathogenic bacteria in this environment is a reality

A mastite bovina é uma doença complexa que traz grandes prejuízos ao produtor de leite. A diversidade micro-biana dos solos, bem como a presença de genes de resistência no ambiente, influencia diretamente a manutenção da mastite na fazenda. O objetivo deste trabalho foi analisar a diversidade bacteriana em solos de pastagem de uma propriedade familiar leiteira, detectando enterobactérias que possam estar envolvidas na etiologia da mastite bovina, e detectar genes que codificam betalactamases de amplo espectro nesses solos. Doze amostras de solo, representativas de diferentes áreas da fazenda localizada no município de Barra do Piraí, Rio de Janeiro, foram coletadas em diferentes épocas do ano. O DNA total foi extraído das amostras, o gene foi amplificado por Nested-PCR e, em seguida, os produtos de amplificação foram separados por DGGE (Denaturing Gradient Gel Electrophoresis). Com a DGGE, foi possível construir dendogramas que representavam efetivamente a diversidade bacteriana desses solos. Oito das amostras de solo foram usadas para amplificar os genes que codificam as enzimas betalactamase TEM (gene blaTEM), SHV (gene blaSHV) e CTX (gene blaCTXM). Em três das oito amostras de solo, foi constatada a presença do gene blaSHV. Os genes blaTEM e blaCTX-M não foram detectados em nenhuma das amostras. A detecção de genes que codificam betalactamases de amplo espectro em solos de pastagens de gado leiteiro é preocupante, pois a transferência de material genético entre bactérias patogênicas e não patogênicas nesse ambiente é uma realidade

Molecular, Chemical, and Sensory Attributes Fingerprinting of Self-Induced Anaerobic Fermented Coffees from Different Altitudes and Processing Methods.

Coffee quality is achieved by performing good practices. This study aimed to evaluate coffees from different altitudes fermented with the self-induced anaerobic method (SIAF) and processed via natural (N) and pulped natural (PN). Molecular (PCR-DGGE), chemical (HPLC, ABTS, DPPH, ATR-FTIR, and GC-MS), and sensory analyses were performed. Leuconostoc predominated both processes and all altitudes. Hanseniaspora and Pichia predominated both processes at 800 and 1200 m. Acids were higher in N coffees for all altitudes. Acetic, malic acid and alcohols were the most abundant. Higher sensory scores were obtained in N (mainly at 1400 m-88.13). Floral and spices were perceived in all samples. ABTS capacity in roasted coffee increased with altitude in PN (2685.71, 2724.03, and 3847.14 µM trolox/g); meanwhile, the opposite was observed in N. High sensory scores were obtained in high altitudes. Alcohols and acids in roasted beans increase with altitude. Leuconostoc and Pichia showed potential as future coffee starters.

Further culture-independent characterization of the lactic microbiota of Serro artisanal cheese.

This study aimed to provide a further characterization of the lactic microbiota present in Minas artisanal cheese (MAC) from the Serro region by using culture-independent methods, as a complementary analysis of a previous study. The total DNA extracted from MAC samples (n = 55) was subjected to repetitive extragenic palindromic-PCR (rep-PCR) and PCR-denaturing gradient gel electrophoresis (PCR-DGGE). Rep-PCR analysis showed that core microbiota of Serro MAC was closely related, independent of the production town, farm size, or time of production. The sequencing of PCR-DGGE bands identified the prevalence of Lactococcus lactis in all samples, and Streptococcus salivarius was also identified. Thus, we conclude that when more accurate methods are unavailable, rep-PCR can be used as a culture-independent method to demonstrate if the microbiota is closely related or not among the samples. PCR-DGGE results also matched to the main findings of high-throughput sequencing, previously presented, confirming its confidence to detect the main microbial groups present in the raw milk cheeses.

Influence of oxygen on the vinyl acetate elimination pathway and microbial community structure of methanogenic sludge.

Methanogenic-aerobic coupled processes were used to biological degradation of vinyl acetate (VA) to provide evidence of oxygen role for their complete elimination from different angles. First, physiological characterization of a continuous methanogenic-aerobic reactor fed by VA and glucose (G) showed that by adding G, the VA got 100% hydrolyzed to acetate, and then, by adding 1 mg·L-1 ·d-1 of dissolved oxygen (DO), this acetate got methanized by 40% and aerobically mineralized by 60%. Second, batch assays in the presence and absence of sodium azide suggest that VA at different concentrations was eliminated by both anaerobic and aerobic metabolic pathways, because without azide and in the presence of 1 mg DO·L-1 increased methane and carbon dioxide formation rates at 80% and 75%, respectively. Finally, microbial population dynamics analysis of the reactor by DGGE-sequencing highlighted that Brevibacillus agri (aerobic) and Methanosarcina barkeri (anaerobic) were identified as responsible for VA elimination by up to 98.6%. PRACTITIONER POINTS: Vinyl acetate is removed by simultaneous methanation and aerobic respiration. Methanosarcina barkeri and Brevibacillus agri removed up to 99% of vinyl acetate. DO and VA have a selective effect on the metabolism and population dynamics.

Seasonal Dynamics of the Honey Bee Gut Microbiota in Colonies Under Subtropical Climate : Seasonal Dynamics of Honey Bee Gut Microbiota.

Honey bees (Apis mellifera) provide invaluable benefits for food production and maintenance of biodiversity of natural environments through pollination. They are widely spread across the world, being adapted to different climatic conditions. To survive the winter in cold temperate regions, honey bees developed different strategies including storage of honey and pollen, confinement of individuals during the winter, and an annual cycle of colony growth and reproduction. Under these conditions, winter honey bees experience physiological changes, including changes in immunity and the composition of honey bee gut microbiota. However, under tropical or subtropical climates, the life cycle can experience alterations, i.e., queens lay eggs during almost all the year and new honey bees emerge constantly. In the present study, we characterized nurses' honey bee gut microbiota in colonies under subtropical region through a year, combining qPCR, PCR-DGGE, and 16S rDNA high-throughput sequencing. We also identified environmental variables involved in those changes. Our results showed that under the mentioned conditions, the number of bacteria is stable throughout the year. Diversity of gut microbiota is higher in spring and lower in summer and winter. Gradual changes in compositions occur between seasons: Lactobacillus spp. predominate in spring while Gilliamella apicola and Snodgrasella alvi predominate in summer and winter. Environmental variables (mainly precipitations) affected the composition of the honey bee gut microbiota. Our findings provide new insights into the dynamics of honey bee gut microbiota and may be useful to understand the adaptation of bees to different environmental conditions.

Effects of glyphosate on soil fungal communities: A field study / Efecto del glifosato en las comunidades fúngicas del suelo: estudio a campo

ABSTRACT The driving forces behind many soil processes are microorganisms and they are able to respond immediately to environmental changes. The soil microbial community impacts on many soil properties. More than one-third of the terrestrial ecosystems are semiarid. However, a limited number of studies have been conducted to characterize soil fungal communities in semiarid grasslands, in particular those of agricultural fields. The aim of this study was to explore changes in the diversity and structure of soil fungal communities in semiarid grasslands, after different doses of glyphosate were applied under field conditions. Changes in soil fungal communities were examined using different approaches including culturing, calcofluor white stain and denaturing gradient gel electrophoresis (DGGE). The different approaches complement each other, revealing different aspects of the effect of glyphosate on soil fungal communities. We demonstrated a negative effect of glyphosate on soil fungal biomass at high doses and an early and transitory stimulatory effect on soil fungal biomass. We also found a negative effect of glyphosate on the species richness of cultivable fungi and changes in the molecular structure of soil fungal communities after double doses or long-term glyphosate application. In summary, our findings demonstrate an overall negative effect of glyphosate on soil fungal communities.

RESUMEN Los microorganismos del suelo son los responsables de llevar a cabo la mayoría de los procesos biológicos que ocurren en el suelo, y son capaces de reaccionar ante el estrés ambiental. Más de un tercio de los ecosistemas terrestres son semiáridos. Sin embargo, son escasos los estudios realizados para caracterizar las comunidades fúngicas en suelos agrícolas en ecosistemas semiáridos. El objetivo del presente trabajo fue estudiar los cambios que se producen en la biomasa, la diversidad y la estructura de las comunidades fúngicas del suelo, luego de la aplicación de distintas dosis de glifosato en condiciones de campo. Se emplearon diferentes técnicas incluidas el cultivo, la tinción directa con blanco de calcoflúor y PCR acoplada a electroforesis en geles de gradiente desnaturalizante (DGGE). Las distintas metodologías empleadas se complementan entre sí al detectar cada una distintos aspectos del efecto del glifosato en las comunidades fúngicas del suelo. Se encontró que el glifosato produce un efecto negativo sobre la biomasa fúngica, también se encontró un efecto transitorio estimulante inmediatamente posterior a la aplicación del herbicida. Además, se vio un efecto negativo sobre la riqueza de hongos cultivables, así como también cambios en la estructura molecular de las comunidades luego de aplicaciones repetidas. En conclusión, se demostró un efecto negativo generalizado sobre las comunidades fúngicas del suelo.

Sulfamethoxazole biodegradation and impacts on soil microbial communities in a Bolivian arid high altitude catchment.

The processes controlling antibiotics fate in ecosystems are poorly understood, yet their presence can inhibit bacterial growth and induce the development of bacterial resistance. Sulfamethoxazole (SMX) is one of the most frequently detected sulfonamides in natural environments due to its low metabolism and molecular properties. This work presents pioneering results on SMX biodegradation and impact in high altitude soils (Bolivian Altiplano), allowing a better understanding of the persistence, spread and impact of this antibiotic at the global watershed scale. Our results showed significant dissipation of SMX in relation to its adsorption, hydrolysis and biotransformation. However, biodegradation appears to be lower in these mountain soils than in lowland soils as widely described in the literature. The half-life of SMX ranges from 12 to 346 days in non-sterile soils. In one soil, no biotic degradation was observed, indicating a likely high persistence. Biodegradation was related to OC content and to proximity to urban activities. Regarding the study of the impacts of SMX, the DGGE results were less sensitive than the sequencing. In general, SMX strongly changes the structure and composition of the studied soils at high altitudes, which is comparable to the observations of other authors in lowland soils. The phylum Actinobacter showed high sensitivity to SMX. In contrast, the abundance of ɣ-proteobacteria remained almost unchanged. Soil contamination with SMX did not lead to the development of the studied resistance genes (sul1 and sul2) in soils where they were absent at the beginning of the experiment. Thus, the presence of SMX resistance genes seems to be related to irrigation with wastewater carrying the studied resistance genes.

Effects of glyphosate on soil fungal communities: A field study.

The driving forces behind many soil processes are microorganisms and they are able to respond immediately to environmental changes. The soil microbial community impacts on many soil properties. More than one-third of the terrestrial ecosystems are semiarid. However, a limited number of studies have been conducted to characterize soil fungal communities in semiarid grasslands, in particular those of agricultural fields. The aim of this study was to explore changes in the diversity and structure of soil fungal communities in semiarid grasslands, after different doses of glyphosate were applied under field conditions. Changes in soil fungal communities were examined using different approaches including culturing, calcofluor white stain and denaturing gradient gel electrophoresis (DGGE). The different approaches complement each other, revealing different aspects of the effect of glyphosate on soil fungal communities. We demonstrated a negative effect of glyphosate on soil fungal biomass at high doses and an early and transitory stimulatory effect on soil fungal biomass. We also found a negative effect of glyphosate on the species richness of cultivable fungi and changes in the molecular structure of soil fungal communities after double doses or long-term glyphosate application. In summary, our findings demonstrate an overall negative effect of glyphosate on soil fungal communities.

Exploring the role of healthy dogs as hosts of enterohepatic Helicobacter species using cultivation-dependent and -independent approaches.

Enterohepatic Helicobacter (EHH) species have been increasingly associated with acute gastroenteritis, inflammatory bowel disease and hepatobiliary diseases in humans. However, their host range and transmission routes are poorly understood. Therefore, the aim of this study was to determine the presence of EHH in healthy dogs using both cultivation-dependent and -independent methods. Three hundred and ninety faecal samples from domestic dogs without gastrointestinal symptoms were analysed between June 2018 and July 2019 in Valdivia (South of Chile). Samples were inoculated on selective medium and in parallel were filtrated over an antibiotic-free blood agar. Both media were incubated in a microaerobic atmosphere at 37°C for 7 days. Colonies were identified by PCR and phylogenetic analysis. A subset of 50 samples (half of them positive for EHH by cultivation and the remaining half negative) was analysed by PCR-Denaturing Gradient Gel Electrophoresis (PCR-DGGE) for direct detection. Cultivation method detected EHH in 15.4% (60/390) of the samples, being the most prevalent species H. canis (5.8%, 23/390) and H. canicola (5.1%, 20/390), followed by H. bilis (3.6%, 14/390) and 'H. winghamensis' (1.3%, 5/390). In contrast, PCR-DGGE method detected Helicobacter DNA in almost all (96%, 48/50) tested samples. On the other hand, the method used also allowed to isolate other Campylobacterales, in fact 44.3% (173/390) of the samples were positive for Campylobacter upsaliensis (43.3%, 169/390) followed by C. jejuni (2.0%, 8/390). Moreover, two strains that presented Campylobacter-like morphology were finally identified as Anaerobiospirillum succiniciproducens. Our results indicate that healthy domestic dogs commonly carry EHH and other Campylobacter species. However, further studies are needed to determine whether and how these Helicobacter and Campylobacter species can be transmitted to humans.

Methanotrophic Community Detected by DNA-SIP at Bertioga's Mangrove Area, Southeast Brazil.

Methanotrophic bacteria can use methane as sole carbon and energy source. Its importance in the environment is related to the mitigation of methane emissions from soil and water to the atmosphere. Brazilian mangroves are highly productive, have potential to methane production, and it is inferred that methanotrophic community is of great importance for this ecosystem. The scope of this study was to investigate the functional and taxonomic diversity of methanotrophic bacteria present in the anthropogenic impacted sediments from Bertioga´s mangrove (SP, Brazil). Sediment sample was cultivated with methane and the microbiota actively involved in methane oxidation was identified by DNA-based stable isotope probing (DNA-SIP) using methane as a labeled substrate. After 4 days (96 h) of incubation and consumption of 0.7 mmol of methane, the most active microorganisms were related to methanotrophs Methylomonas and Methylobacter as well as to methylotrophic Methylotenera, indicating a possible association of these bacterial groups within a methane-derived food chain in the Bertioga mangrove. The abundance of genera Methylomonas, able to couple methane oxidation to nitrate reduction, may indicate that under low dissolved oxygen tensions, some aerobic methanotrophs could shift to intraerobic methane oxidation to avoid oxygen starvation.

Genetic diversity of N-fixing and plant growth-promoting bacterial community in different sugarcane genotypes, association habitat and phenological phase of the crop.

This study aimed to evaluate the genetic diversity of bacterial community associated to different sugarcane genotypes, association habitat and phenological phase of the culture, as well as to isolate, to identify and to characterize your potential for plant growth-promoting. Root and rhizospheric soil samples from RB 92579 and RB 867515 varieties were collected at 120 and 300 days after regrowth (DAR). The diversity of bacterial was evaluated through of the 16S rRNA and nifH genes. We found greater genetic diversity in the root endophytic habitat at 120 DAR. We identify the genera Burkholderia sp., Pantoea sp., Erwinia sp., Stenotrophomonas sp., Enterobacter sp. and Pseudomonas sp. The genera Bacillus sp. and Dyella sp. were only identified in the variety RB 92579. We found indices above 50% for biological nitrogen fixation, production of indole acetic acid and phosphate solubilization, showing that the use of these bacteria in biotechnological products is very promising.

Evaluation of nitrogen removal and the microbial community in a submerged aerated biological filter (SABF), secondary decanters (SD), and horizontal subsurface flow constructed wetlands (HSSF-CW) for the treatment of kennel effluent.

To ensure microbial activity and a reaction equilibrium with efficiency and energy saving, it is important to know the factors that influence microbiological nitrogen removal in wastewater. Thus, it was investigated the microorganisms and their products involved in the treatment of kennel effluents operated with different aeration times, phase 1 (7 h of continuous daily aeration), phase 2 (5 h of continuous daily aeration), and phase 3 (intermittent aeration every 2 h), monitoring chemical and physical parameters weekly, monthly microbiological, and qualitative and quantitative microbiological analyzes at the end of each applied aeration phase. The results showed a higher mean growth of nitrifying bacteria (NB) (106) and denitrifying bacteria (DB) (1022) in phase with intermittent aeration, in which better total nitrogen (TN) removal performance, with 33%, was achieved, against 21% in phase 1 and 17% in phase 2, due to the longer aeration time and lower carbon/nitrogen ratio (15.7), compared with the other phases. The presence of ammonia-oxidizing bacteria (AOB), the genus Nitrobacter nitrite-oxidizing bacteria (NOB), and DB were detected by PCR with specific primers at all phases. The analysis performed by 16S-rRNA DGGE revealed the genres Thauera at all phases; Betaproteobacteria and Acidovorax in phase 3; Azoarcus in phases 2 and 3; Clostridium, Bacillus, Lactobacillus, Turicibacter, Rhodopseudomonas, and Saccharibacteria in phase 1, which are related to the nitrogen removal, most of them by denitrifying. It is concluded that, with the characterization of the microbial community and the analysis of nitrogen compounds, it was determined, consistently, that the studied treatment system has microbiological capacity to remove TN, with the phase 3 aeration strategy, by simultaneous nitrification and denitrification (SND). Due to the high density of DB, most of the nitrification occurred by heterotrophic nitrification-aerobic. And denitrification occurred by heterotrophic and autotrophic forms, since the higher rate of oxygen application did not harm the DB. Therefore, the aeration and carbon conditions in phase 3 favored the activity of the microorganisms involved in these different routes. It is considered that, in order to increase autotrophic nitrification-aerobic, it is necessary to exhaust the volume of sludge in the secondary settlers (SD), further reducing the carbon/nitrogen ratio, through more frequent cleaning, whose periodicity should be the object of further studies. Graphical abstract.

Anaerobic digestion of aqueous phase from hydrothermal liquefaction of Spirulina using biostimulated sludge.

Hydrothermal liquefaction is a process that converts wet biomass into biofuels, more specifically bio-crude oil. During the process, post hydrothermal liquefaction waste water (PHWW) is generated, rich in nutrient and organic matter, however potentially toxic. Anaerobic digestion of PHWW from Spirulina, was evaluated using biostimulated sludge as a strategy to optimize the process. The biostimulation was conducted in a sequential batch reactor fed with organic acids and methanol aiming at development of acetogenic and methanogenic microorganism. Anaerobic biodegradability batch assays were performed, with biostimulated sludge and with non-biostimulated sludge, using increasing PHWW concentrations. Biostimulated sludge were more favourable for reaching higher methane yields at higher organic matter concentrations in comparison to non-biostimulated sludge, presenting less inhibition at conditions tested. Biostimulation was a key process to select and favour potential microorganisms involved in specialized uptake of recalcitrant compounds, such as Mesotoga and Methanomethylovorans.

Root colonization and arbuscular mycorrhizal fungal community composition in a genetically modified maize, its non-modified isoline, and a landrace.

The use of genetically modified (GM) plants has increased in recent decades, but there are uncertainties about their effects on soil microbial communities. Aiming to quantify root colonization and characterize arbuscular mycorrhizal fungi (AMF) communities associated with roots and rhizosphere soil of different maize genotypes, a field trial was carried out in Southern Brazil with three maize genotypes as follows: a GM hybrid (DKB 240 VTPRO), its non-modified isoline (DKB 240), and a landrace (Pixurum). Soil samples were collected to evaluate the occurrence of AMF during the growth of corn genotypes at sowing and V3 (vegetative), R1 (flowering), and R3 (grain formation) stages of the crop. The occurrence of AMF was determined by the morphological identification of spores, and by analyzing AMF community composition in soil and roots of maize, using PCR-DGGE. The GM genotype of maize promoted lower mycorrhizal colonization in the vegetative stage and had lower sporulation at grain development than the conventional hybrid and the landrace maize. Twenty AMF morphotypes were identified and 13 were associated with all maize genotypes. The genera Acaulospora, Glomus, and Dentiscutata had the largest numbers of species. There were no differences in AMF community composition due to maize genotypes or genetic modification, but crop phenological stages affected AMF communities associated with maize roots.

Yeast and bacterial diversity, dynamics and fermentative kinetics during small-scale tequila spontaneous fermentation.

The study of microbial communities associated with spontaneous fermentation of agave juice for tequila production is required to develop starter cultures that improve both yield and quality of the final product. Quantification by HPLC of primary metabolites produced during the fermentations was determined. A polyphasic approach using plate count, isolation and identification of microorganisms, denaturing gradient gel electrophoresis and next generation sequencing was carried out to describe the diversity and dynamics of yeasts and bacteria during small-scale spontaneous fermentations of agave juice from two-year samplings. High heterogeneity in microbial populations and fermentation parameters were observed, with bacteria showing higher diversity than yeast. The core microorganisms identified were Saccharomyces cerevisiae and Lactobacillus fermentum. Practices in tequila production changed during the two-year period, which affected microbial community structure and the time to end fermentation. Bacterial growth and concomitant lactic acid production were associated with low ethanol production, thus bacteria could be defined as contaminants in tequila fermentation and efforts to control them should be implemented.

Controlling methane and hydrogen production from cheese whey in an EGSB reactor by changing the HRT.

This study assessed the effects of hydraulic retention time (HRT; 8 h-0.25 h) on simultaneous hydrogen and methane production from cheese whey (5000 mg carbohydrates/L) in a mesophilic (30 °C) expanded granular sludge bed (EGSB) reactor. Methane production was observed at HRTs from 4 to 0.25 h. The maximum methane yield (9.8 ± 1.9 mL CH4/g CODap, reported as milliliter CH4 per gram of COD applied) and methane production rate (461 ± 75 mL CH4/day Lreactor) occurred at HRTs of 4 h and 2 h, respectively. Hydrogen production increased as methane production decreased with decreasing HRT from 8 to 0.25 h. The maximum hydrogen yield of 3.2 ± 0.3 mL H2/g CODap (reported as mL H2 per gram of COD applied) and hydrogen production rate of 1951 ± 171 mL H2/day Lreactor were observed at the HRT of 0.25 h. The decrease in HRT from 8 to 0.25 h caused larger changes in the bacterial populations than the archaea populations. With the decrease in HRT (6 h-0.25 h), the Shannon diversity index decreased (3.02-2.87) for bacteria and increased (1.49-1.83) for archaea. The bacterial dominance increased (0.059-0.066) as the archaea dominance decreased (0.292-0.201) with the HRT decrease from 6 to 0.25 h.

Bacterial community structure in soils contaminated with electronic waste pollutants from Delhi NCR, India

Background: Microbial community analysis of electronic waste (e-waste)-polluted environments is of interest to understand the effect of toxic e-waste pollutants on the soil microbial community and to evaluate novel microorganisms resisting the toxic environment. The present study aims to investigate the bacterial community structure in soils contaminated with e-waste from various sites of Loni and Mandoli (National Capital Region (NCR), India) where e-waste dumping and recycling activities are being carried out for many years. Results: Interferences to soil metagenomic DNA extraction and PCR amplification were observed because of the presence of inhibiting components derived from circuit boards. Whole-metagenome sequencing on the Illumina MiSeq platform showed that the most abundant phyla were Proteobacteria and Firmicutes. Deltaproteobacteria and Betaproteobacteria were the most common classes under Proteobacteria. Denaturing gradient gel electrophoresis (DGGE) analysis of the bacterial 16S rRNA gene showed that e-waste contamination altered the soil bacterial composition and diversity. There was a decrease in the number of predominant bacterial groups like Proteobacteria and Firmicutes but emergence of Actinobacteria in the contaminated soil samples. Conclusions: This is the first report describing the bacterial community structure of composite soil samples of ewaste-contaminated sites of Loni and Mandoli, Delhi NCR, India. The findings indicate that novel bacteria with potential bioremediating properties may be present in the e-waste-contaminated sites and hence need to be evaluated further.