Model-aided targeted volatile fatty acid production from food waste using a defined co-culture microbial community.

The production of volatile fatty acids (VFA) is gaining momentum due to their central role in the emerging carboxylate platform. Particularly, the production of the longest VFA (from butyrate to caproate) is desired due to their increased economic value and easier downstream processing. While the use of undefined microbial cultures is usually preferred with organic waste streams, the use of defined microbial co-culture processes could tackle some of their drawbacks such as poor control over the process outcome, which often leads to low selectivity for the desired products. However, the extensive experimentation needed to design a co-culture system hinders the use of this technology. In this work, a workflow based on the combined use of mathematical models and wet experimentation is proposed to accelerate the design of novel bioprocesses. In particular, a co-culture consisting of Pediococcus pentosaceus and Megaphaera cerevisiae is used to target the production of high-value odd- and even­carbon VFA. An unstructured kinetic model was developed, calibrated and used to design experiments with the goal of increasing the selectivity for the desired VFA, which were experimentally validated. In the case of even­carbon VFA, the experimental validation showed an increase of 38 % in caproate yield and, in the case of enhanced odd­carbon VFA experiments, the yield of butyrate and caproate diminished by 62 % and 94 %, respectively, while propionate became one of the main end products and valerate yield value increased from 0.007 to 0.085 gvalearte per gconsumed sugar. The workflow followed in this work proved to be a sound tool for bioprocess design due to its capacity to explore and design new experiments in silico in a fast way and ability to quickly adapt to new scenarios.

Ischnura praematura sp. nov. (Odonata: Zygoptera: Coenagrionidae): a species from Yunnan (China) whose females mate in the teneral state.

Ischnura praematura sp. nov. (Holotype , China, Yunnan, Lijiang, 263103.54N, 1001338.89E, 2396 m, 04 xii 2015, I. Sanmartn-Villar H. Zhang leg.) is morphologically described, illustrated and compared with close species of the genus. Ischnura praematura can be mainly distinguished from its congeners I. aurora, I. rubilio and I. asiatica by its abdominal and thoracic morphology and colouration. The posterior lobe of the prothorax is elevated in I. praematura and the mesostigmal plates possess dorsal triangular protuberances. Ischnura praematura shows pointed paraprocts, internalised wide cerci and lacks a dorsal tuberculum in the tenth abdominal segment. The blue abdominal colouration is present in the last three segments (incomplete for segment eight and ten in some individuals). No female polychromatism was detected and all females observed possessed different colouration than male (gynochrome). Morphological distinctiveness of the species is supported by genetic analyses, which show that I. praematura forms a well-supported, monophyletic clade, with I. asiatica, I. ezoin and I. pumilio as the most closely related species. In the field, mature females show strong reluctance to mate, and males were observed mating with newly emerged females.

Incidence and Diversity of Torix Rickettsia-Odonata Symbioses.

Heritable microbes are an important component of invertebrate biology, acting both as beneficial symbionts and reproductive parasites. Whilst most previous research has focussed on the 'Wolbachia pandemic', recent work has emphasised the importance of other microbial symbionts. In this study, we present a survey of odonates (dragonflies and damselflies) for torix group Rickettsia, following previous research indicating that this clade can be common in other aquatic insect groups. PCR assays were used to screen a broad range of odonates from two continents and revealed 8 of 76 species tested were infected with Rickettsia. We then conducted further deeper screening of UK representatives of the Coenagrionidae damselfly family, revealing 6 of 8 UK coenagrionid species to be positive for torix Rickettsia. Analysis of Rickettsia gene sequences supported multiple establishments of symbiosis in the group. Some strains were shared between UK coenagrionid species that shared mtDNA barcodes, indicating a likely route for mitochondrial introgression between sister species. There was also evidence of coinfecting Rickettsia strains in two species. FISH analysis indicated Rickettsia were observed in the ovarioles, consistent with heritable symbiosis. We conclude that torix Rickettsia represent an important associate of odonates, being found in a broad range of species from both Europe and South America. There is evidence that coinfection can occur, vertical transmission is likely, and that symbiont movement following hybridisation may underpin the lack of 'barcoding gap' between well-established species pairs in the genus. Future work should establish the biological significance of the symbioses observed.

Steering the conversion of protein residues to volatile fatty acids by adjusting pH.

This study investigates the influence of pH on protein conversion into volatile fatty acids by anaerobic mixed-culture fermentation, a topic that, in contrast to glucose fermentation, only had scarce and contradictory information available. Several experiments were performed with two model proteins (casein and gelatin) at three different pH values (5, 7 and 9) using chemostats and batch tests. Highest conversion was reached at neutral pH although complete acidification was never achieved. Longer chain carboxylates production was favoured at low pH, while acetic acid was the main product at pH 7 and 9. Amino acids preferential consumption also varied with pH and protein composition. In fact, protein conversion stoichiometry is mainly driven by energetic yields and amino acid molecular configuration. Overall, this study identifies pH adjustment as a way to steer volatile fatty acid production during mixed-culture fermentation of proteins.

Microbial invasions in sludge anaerobic digesters.

Among processes that control microbial community assembly, microbial invasion has received little attention until recently, especially in the field of anaerobic digestion. However, knowledge of the principles regulating the taxonomic and functional stability of microbial communities is key to truly develop better predictive models and effective management strategies for the anaerobic digestion process. To date, available studies focus on microbial invasions in digesters feed with activated sludge from municipal wastewater treatment plants. Herein, this review summarizes the importance of invasions for anaerobic digestion management, the ecological theories about microbial invasions, the traits of activated sludge microorganisms entering the digesters, and the resident communities of anaerobic reactors that are relevant for invasions and the current knowledge about the success and impacts of invasions, and discusses the research needs on this topic. The initial data indicate that the impact of invasions is low and only a small percentage of the mostly aerobic microorganisms present in the activated sludge feed are able to become stablished in the anaerobic digesters. However, there are still numerous unknowns about microbial invasions in anaerobic digestion including the influence of anaerobic feedstocks or process perturbances that new approaches on microbial ecology could unveil. KEY POINTS: • Microbial invasions are key processes to develop better strategies for digesters management. • Knowledge on pathogen invasions can improve anaerobic digestion microbial safety. • To date, the number of successful invasions on anaerobic digesters from activated sludge organisms is low. • Feed organisms detected in digesters are mostly inactive residual populations. • Need to expand the range of invaders and operational scenarios studied.

Widespread Wolbachia infection in an insular radiation of damselflies (Odonata, Coenagrionidae).

Wolbachia is one of the most common endosymbionts found infecting arthropods. Theory predicts symbionts like Wolbachia will be more common in species radiations, as host shift events occur with greatest frequency between closely related species. Further, the presence of Wolbachia itself may engender reproductive isolation, and promote speciation of their hosts. Here we screened 178 individuals belonging to 30 species of the damselfly genera Nesobasis and Melanesobasis - species radiations endemic to the Fiji archipelago in the South Pacific - for Wolbachia, using multilocus sequence typing to characterize bacterial strains. Incidence of Wolbachia was 71% in Nesobasis and 40% in Melanesobasis, and prevalence was also high, with an average of 88% in the Nesobasis species screened. We identified a total of 25 Wolbachia strains, belonging to supergroups A, B and F, with some epidemic strains present in multiple species. The occurrence of Wolbachia in both males and females, and the similar global prevalence found in both sexes rules out any strong effect of Wolbachia on the primary sex-ratio, but are compatible with the phenotype of cytoplasmic incompatibility. Nesobasis has higher species richness than most endemic island damselfly genera, and we discuss the potential for endosymbiont-mediated speciation within this group.

Mechanisms of allopatric speciation in an Antillean damselfly genus (Odonata, Zygoptera): Vicariance or long-distance dispersal?

We have examined divergence times of the Antillean damselfly genus Hypolestes, to elucidate which mechanism of allopatric speciation, vicariance or long-distance dispersal, could better explain the currently observed disjunct distributions of this genus. Samples of the three extant species of the genus, Hypolestes clara (Jamaica), H. hatuey (Hispaniola) and H. trinitatis (Cuba), were collected. Mitochondrial and nuclear DNA gene fragments were amplified to reconstruct phylogenetic relationships and estimate divergence times in this genus. Hypolestes comprises currently three species, which consist in four geographically and genetically isolated lineages located in Jamaica, Hispaniola, Eastern Cuba and Central Cuba. Results of our analyses suggest that the three species diverged between ∼5.91 and 1.69 mya, and that the separation between the lineages from Central Cuba and Eastern Cuba occurred between ∼2.0 and 0.62 mya. Disjunct distributions in the genus Hypolestes can be better explained by a long-distance dispersal mechanism, since the divergence times of the three species do not coincide with the timeline formation of the geographic barriers between Cuba, Hispaniola and Jamaica. The Cuban lineages of H. trinitatis constitute different molecular operational taxonomic units (MOTU). The elevation of these MOTU to the species category requires the analysis of additional characters.

The time response of anaerobic digestion microbiome during an organic loading rate shock.

Knowledge of connections between operational conditions, process stability, and microbial community dynamics is essential to enhance anaerobic digestion (AD) process efficiency and management. In this study, the detailed temporal effects of a sudden glycerol-based organic overloading on the AD microbial community and process imbalance were investigated in two replicate anaerobic digesters by a time-intensive sampling scheme. The microbial community time response to the overloading event was shorter than the shifts of reactor performance parameters. An increase in bacterial community dynamics and in the abundances of several microbial taxa, mainly within the Firmicutes, Tenericutes, and Chloroflexi phyla and Methanoculleus genera, could be detected prior to any shift on the reactor operational parameters. Reactor acidification already started within the first 24 h of the shock and headed the AD process to total inhibition in 72 h alongside with the largest shifts on microbiome, mostly the increase of Anaerosinus sp. and hydrogenotrophic methanogenic Archaea. In sum, this work proved that AD microbial community reacts very quickly to an organic overloading and some shifts occur prior to alterations on the performance parameters. The latter is very interesting as it can be used to improve AD process management protocols.

Environmental assessment of alternative treatment schemes for energy and nutrient recovery from livestock manure.

The application of livestock manure on agricultural land is being restricted due to its significant content of phosphorus (P) and nitrogen (N), leading to eutrophication. At the same time, the growing demand for N and P mineral fertilizers is increasing their production costs and causing the depletion of natural phosphate rock deposits. In the present work, seven technologically feasible treatment schemes for energy (biogas) and nutrient recovery (e.g., struvite precipitation) and/or removal (e.g., partial nitritation/anammox) were evaluated from an environmental perspective. In general, while approaches based solely on energy recovery and use of digestate as fertilizer are commonly limited by community regulations, strategies pursuing the generation of high-quality struvite are not environmentally sound alternatives. In contrast, schemes that include further solid/liquid separation of the digestate improved the environmental profile, and their combination with an additional N-removal stage would lead to the most environmental-friendly framework. However, the preferred scenario was identified to be highly dependent on the particular conditions of each site, integrating environmental, social and economic criteria.

What happens with organic micropollutants during UV disinfection in WWTPs? A global perspective from laboratory to full-scale.

The phototransformation of 18 organic micropollutants (OMPs) commonly detected in wastewater treatment plant (WWTP) effluents was examined attempting to explain their fate during UV disinfection in WWTPs. For this purpose, a lab-scale UV reactor (lamp emitting at 254nm) was used to study the influence of the operational conditions (UV dose, temperature and water matrix) on OMPs abatement and disinfection efficiency. Chemical properties of OMPs and the quality of treated effluent were identified as key factors affecting the phototransformation rate of these compounds. Sampling campaigns were carried out at the inlet and outlet of UV systems of three WWTPs, and the results evidenced that only the most photosensitive compounds, such as sulfamethoxazole and diclofenac, are eliminated. Therefore, despite UV treatment is an effective technology to phototransform OMPs, the UV doses typically applied for disinfection (10-50mJ/cm2) are not sufficient to remove them. Consequently, small modifications (increase of UV dose, use of catalysts) should be applied in WWTPs to enhance the abatement of OMPs in UV systems.

Odonata (dragonflies and damselflies) as a bridge between ecology and evolutionary genomics.

Odonata (dragonflies and damselflies) present an unparalleled insect model to integrate evolutionary genomics with ecology for the study of insect evolution. Key features of Odonata include their ancient phylogenetic position, extensive phenotypic and ecological diversity, several unique evolutionary innovations, ease of study in the wild and usefulness as bioindicators for freshwater ecosystems worldwide. In this review, we synthesize studies on the evolution, ecology and physiology of odonates, highlighting those areas where the integration of ecology with genomics would yield significant insights into the evolutionary processes that would not be gained easily by working on other animal groups. We argue that the unique features of this group combined with their complex life cycle, flight behaviour, diversity in ecological niches and their sensitivity to anthropogenic change make odonates a promising and fruitful taxon for genomics focused research. Future areas of research that deserve increased attention are also briefly outlined.

Is anaerobic digestion effective for the removal of organic micropollutants and biological activities from sewage sludge?

The occurrence of emerging organic micropollutants (OMPs) in sewage sludge has been widely reported; nevertheless, their fate during sludge treatment remains unclear. The objective of this work was to study the fate of OMPs during mesophilic and thermophilic anaerobic digestion (AD), the most common processes used for sludge stabilization, by using raw sewage sludge without spiking OMPs. Moreover, the results of analytical chemistry were complemented with biological assays in order to verify the possible adverse effects (estrogenic and genotoxic) on the environment and human health in view of an agricultural (re)use of digested sludge. Musk fragrances (AHTN, HHCB), ibuprofen (IBP) and triclosan (TCS) were the most abundant compounds detected in sewage sludge. In general, the efficiency of the AD process was not dependent on operational parameters but compound-specific: some OMPs were highly biotransformed (e.g. sulfamethoxazole and naproxen), while others were only slightly affected (e.g. IBP and TCS) or even unaltered (e.g. AHTN and HHCB). The MCF-7 assay evidenced that estrogenicity removal was driven by temperature. The Ames test did not show point mutation in Salmonella typhimurium while the Comet test exhibited a genotoxic effect on human leukocytes attenuated by AD. This study highlights the importance of combining chemical analysis and biological activities in order to establish appropriate operational strategies for a safer disposal of sewage sludge. Actually, it was demonstrated that temperature has an insignificant effect on the disappearance of the parent compounds while it is crucial to decrease estrogenicity.

Effect of oxygen on the microbial activities of thermophilic anaerobic biomass.

Low oxygen levels (µgO2L(-1)) in anaerobic reactors are quite common and no relevant consequences are expected. On the contrary, higher concentrations could affect the process. This work aimed to study the influence of oxygen (4.3 and 8.8mgO2L(-1), respectively) on the different microbial activities (hydrolytic, acidogenic and methanogenic) of thermophilic anaerobic biomass and on the methanogenic community structure. Batch tests in presence of oxygen were conducted using specific substrates for each biological activity and a blank (with minimum oxygen) was included. No effect of oxygen was observed on the hydrolytic and acidogenic activities. In contrast, the methane production rate decreased by 40% in all oxygenated batches and the development of active archaeal community was slower in presence of 8.8mgO2L(-1). However, despite this sensitivity of methanogens to oxygen at saturation levels, the inhibition was reversible.

Status of hormones and painkillers in wastewater effluents across several European states-considerations for the EU watch list concerning estradiols and diclofenac.

Present technologies for wastewater treatment do not sufficiently address the increasing pollution situation of receiving water bodies, especially with the growing use of personal care products and pharmaceuticals (PPCP) in the private household and health sector. The relevance of addressing this problem of organic pollutants was taken into account by the Directive 2013/39/EU that introduced (i) the quality evaluation of aquatic compartments, (ii) the polluter pays principle, (iii) the need for innovative and affordable wastewater treatment technologies, and (iv) the identification of pollution causes including a list of principal compounds to be monitored. In addition, a watch list of 10 other substances was recently defined by Decision 2015/495 on March 20, 2015. This list contains, among several recalcitrant chemicals, the painkiller diclofenac and the hormones 17ß-estradiol and 17α-ethinylestradiol. Although some modern approaches for their removal exist, such as advanced oxidation processes (AOPs), retrofitting most wastewater treatment plants with AOPs will not be acceptable as consistent investment at reasonable operational cost. Additionally, by-product and transformation product formation has to be considered. The same is true for membrane-based technologies (nanofiltration, reversed osmosis) despite of the incredible progress that has been made during recent years, because these systems lead to higher operation costs (mainly due to higher energy consumption) so that the majority of communities will not easily accept them. Advanced technologies in wastewater treatment like membrane bioreactors (MBR) that integrate biological degradation of organic matter with membrane filtration have proven a more complete elimination of emerging pollutants in a rather cost- and labor-intensive technology. Still, most of the presently applied methods are incapable of removing critical compounds completely. In this opinion paper, the state of the art of European WWTPs is reflected, and capacities of single methods are described. Furthermore, the need for analytical standards, risk assessment, and economic planning is stressed. The survey results in the conclusion that combinations of different conventional and advanced technologies including biological and plant-based strategies seem to be most promising to solve the burning problem of polluting our environment with hazardous emerging xenobiotics.

Understanding the fate of organic micropollutants in sand and granular activated carbon biofiltration systems.

In this study, sand and granular activated carbon (GAC) biofilters were comparatively assessed as post-treatment technologies of secondary effluents, including the fate of 18 organic micropollutants (OMPs). To determine the contribution of adsorption and biotransformation in OMP removal, four reactors were operated (two biofilters (with biological activity) and two filters (without biological activity)). In addition, the influence of empty bed contact time (EBCT), ranging from 0.012 to 3.2d, and type of secondary effluent (anaerobic and aerobic) were evaluated. Organic matter, ammonium and nitrate were removed in both biofilters, being their adsorption higher on GAC than on sand. According to the behaviour exhibited, OMPs were classified in three different categories: I) biotransformation and high adsorption on GAC and sand (galaxolide, tonalide, celestolide and triclosan), II) biotransformation, high adsorption on GAC but low or null adsorption on sand (ibuprofen, naproxen, fluoxetine, erythromycin, roxythromycim, sulfamethoxazole, trimethoprim, bisphenol A, estrone, 17ß-estradiol and 17α-ethinylestradiol), and, III) only adsorption on GAC (carbamazepine, diazepam and diclofenac). No influence of EBCT (in the range tested) and type of secondary effluent was observed in GAC reactors, whereas saturation and kinetic limitation of biotransformation were observed in sand reactors. Taking into account that most of the organic micropollutants studied (around 60%) fell into category II, biotransformation is crucial for the elimination of OMPs in sand biofilters.

Biotransformation of pharmaceuticals under nitrification, nitratation and heterotrophic conditions.

The effect of nitrification, nitratation and heterotrophic conditions on the biotransformation of several pharmaceuticals in a highly enriched nitrifying activated sludge was evaluated in this study by selective activation of ammonia oxidizing bacteria (AOB), nitrite oxidizing bacteria (NOB) and heterotrophic bacteria. Nitrifiers displayed a noticeable capacity to process ibuprofen due to hydroxylation by ammonia monooxygenase (AMO) to produce 2-hydroxy-ibuprofen. Naproxen was also biotransformed under nitrifying conditions. On the other hand, heterotrophic bacteria present in the nitrifying activated sludge (NAS) biotransformed sulfamethoxazole. In contrast, both nitrifying and heterotrophic activities were ineffective against diclofenac, diazepam, carbamazepine and trimethoprim. Similar biotransformation rates of erythromycin, roxithromycin and fluoxetine were observed under all conditions tested. Overall, results from this study give more evidence on the role of the different microbial communities present in activated sludge reactors on the biological removal of pharmaceuticals.

Hypolestes hatuey sp. nov.: a new species of the enigmatic genus Hypolestes (Odonata, Hypolestidae) from Hispaniola.

Both sexes of Hypolestes hatuey Torres-Cambas, sp. nov. (Odonata: Zygoptera: Hypolestidae) from Hispaniola are described and illustrated here. This newly described species differs from H. trinitatis and H. clara, the other two species within the genus, by the morphology of the genital ligula and male cerci. Females of H. hatuey sp. nov. differ from H. clara by the shape of the female antehumeral stripe and wing venation. Morphological distinctiveness in males is supported by genetic differences in the 16S mitochondrial gene. Following the categories and criteria of the IUCN Red List of Threatened Species, we suggest this species should be listed as Data Deficient (DD), given that available data on its distribution are too limited to assess its risk of extinction.

Modelling cometabolic biotransformation of organic micropollutants in nitrifying reactors.

Cometabolism is the ability of microorganisms to degrade non-growth substrates in the presence of primary substrates, being the main removal mechanism behind the biotransformation of organic micropollutants in wastewater treatment plants. In this paper, a cometabolic Monod-type kinetics, linking biotransformation of micropollutants with primary substrate degradation, was applied to a highly enriched nitrifying activated sludge (NAS) reactor operated under different operational conditions (hydraulic retention time (HRT) and nitrifying activity). A dynamic model of the bioreactor was built taking into account biotransformation, sorption and volatilization. The micropollutant transformation capacity (Tc), the half-saturation constant (Ksc) and the solid-liquid partitioning coefficient (Kd) of several organic micropollutants were estimated at 25 °C using an optimization algorithm to fit experimental data to the proposed model with the cometabolic Monod-type biotransformation kinetics. The cometabolic Monod-type kinetic model was validated under different HRTs (1.0-3.7 d) and nitrification rates (0.12-0.45 g N/g VSS d), describing more accurately the fate of those compounds affected by the biological activity of nitrifiers (ibuprofen, naproxen, erythromycin and roxithromycin) compared to the commonly applied pseudo-first order micropollutant biotransformation kinetics, which does not link biotransformation of micropollutants to consumption of primary substrate. Furthermore, in contrast to the pseudo-first order biotransformation constant (k(biol)), the proposed cometabolic kinetic coefficients are independent of operational conditions such as the nitrogen loading rate applied. Also, the influence of the kinetic parameters on the biotransformation efficiency of NAS reactors, defined as the relative amount of the total inlet micropollutant load being biotransformed, was assessed considering different HRTs and nitrification rates.

Next generation sequencing yields the complete mitochondrial genome of the scarce blue-tailed damselfly, Ischnura pumilio.

We report the entire mitochondrial genome of the scarce blue-tailed damselfly, Ischnura pumilio (Odonata, Coenagrionidae), using next-generation sequencing on genomic DNA. A de novo assembly provided a single contiguous sequence of 15,250 bp that contained the A + T-rich region and all standard coding regions; gene configuration is similar to other odonates and comprises 13 protein-coding genes, two rRNA genes (12 S and 16 S rRNA) and 22 tRNA genes. We found a unique intergenic spacer in I. pumilio and confirm that the intergenic spacer s5 likely represents a synapomorphy between Anisoptera and Zygoptera. This is the first mitogenome sequence obtained for a member of the Coenagrionidae and demonstrates how next-generation sequencing technology can obtain mtDNA genome sequences without prior sample processing or primer design.

Relationship between phenol degradation efficiency and microbial community structure in an anaerobic SBR.

Phenol is a common wastewater contaminant from various industrial processes, including petrochemical refineries and chemical compounds production. Due to its toxicity to microbial activity, it can affect the efficiency of biological wastewater treatment processes. In this study, the efficiency of an Anaerobic Sequencing Batch Reactor (ASBR) fed with increasing phenol concentrations (from 120 to 1200 mg L(-1)) was assessed and the relationship between phenol degradation capacity and the microbial community structure was evaluated. Up to a feeding concentration of 800 mg L(-1), the initial degradation rate steadily increased with phenol concentration (up to 180 mg L(-1) d(-1)) and the elimination capacity remained relatively constant around 27 mg phenol removed∙gVSS(-1) d(-1). Operation at higher concentrations (1200 mg L(-1)) resulted in a still efficient but slower process: the elimination capacity and the initial degradation rate decreased to, respectively, 11 mg phenol removed∙gVSS(-1) d(-1) and 154 mg L(-1) d(-1). As revealed by Denaturing Gradient Gel Electrophoresis (DGGE) analysis, the increase of phenol concentration induced level-dependent structural modifications of the community composition which suggest an adaptation process. The increase of phenol concentration from 120 to 800 mg L(-1) had little effect on the community structure, while it involved drastic structural changes when increasing from 800 to 1200 mg L(-1), including a strong community structure shift, suggesting the specialization of the community through the emergence and selection of most adapted phylotypes. The thresholds of structural and functional disturbances were similar, suggesting the correlation of degradation performance and community structure. The Canonical Correspondence Analysis (CCA) confirmed that the ASBR functional performance was essentially driven by specific community traits. Under the highest feeding concentration, the most abundant ribotype probably involved in successful phenol degradation at 1200 mg L(-1) was affiliated to the Anaerolineaceae family.