Polyhydroxyalkanoates production from a waste-derived feedstock driven by the reactor operating conditions: the role of biomass microbiome and its reactivation capacity

In the last years, the production of plastics at large-scale has been an issue of great concern due to their environmental and health impacts. Biobased and biodegradable plastics, such as polyhydroxyalkanoates (PHA), have emerged as a possible more ecological and sustainable alternative to those conventional plastics. PHA can be produced by mixed microbial cultures (MMC) from waste-based feedstocks. This work aimed to maximize the PHA production from fermented brewer's spent grain by selecting an efficient PHA-accumulating MMC, applying different organic loading rates (OLR, 45 or 60 CmmolFP/(L.d)) and sludge retention times (SRT, 4 or 2 days). Additionally, the biomass reactivation capacity after storage was evaluated. The global PHA productivity depends on the operating conditions, achieving its maximum (3.55 ± 0.8 gPHA/(L.d)) at the highest OLR and SRT tested. After storage, the global PHA productivity was similar to the one achieved before the starvation period. Depending on the operating conditions, the microbiome has its own bacterial composition signature containing various PHA-producers genera (e.g., Acidovorax, Amaricoccus, Brevundimonas, Leucobacter, Leadbetterella and Thauera) that persisted over time, albeit at different relative abundancies. A comprehensive overview of PHA production with data on system performance and microbiome is presented, which thereby can further contribute to the upscale of such valorisation processes.info:eu-repo/semantics/publishedVersio

Acidogenic Fermentation of Brewers’ Spent Grain Monitored through Two-Dimensional Fluorescence Spectroscopy

Funding Information: This research was funded by national funds from the FCT─Fundação para a Ciência e a Tecnologia, I.P. through the project PTDC/BTA-BTA/31746/2017, in the scope of projects UIDP/04378/2020 and UIDB/04378/2020 of the Research Unit on Applied Molecular Biosciences─UCIBIO, project LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy─i4HB, and the project UIDB/50016/2020 of the Associate Laboratory Centro de Biotecnologia e Química Fina─CBQF. This research was also funded by the Associate Laboratory for Green Chemistry─LAQV, which is financed by national funds from FCT/MCTES (UIDB/50006/2020 and UIDP/50006/2020). E.C.G. gratefully acknowledges FCT for grant SFRH/BD/136300/2018. Publisher Copyright: © 2023 The Authors. Published by American Chemical Society.Biological systems are commonly controlled and monitored through offline and time-consuming tools, which often impairs an effective and real-time response to counteract system disturbances. The feasibility of using two-dimensional (2D) fluorescence spectroscopy as a non-invasive, non-destructive, and real-time procedure to monitor the acidogenic fermentation of brewer’s spent grain (BSG) in a granular sludge reactor was evaluated. For that, the effect of pH fluctuations on the system response was used as a model to ascertain the 2D fluorescence spectroscopy applicability to monitor the process performance, namely, to predict the fermentation products (FP) and the soluble protein (SProt) concentrations in the effluent stream through mathematical analysis. The pH fluctuations over the course of the reactor’s operation altered the granules’ microbiome composition, leading to different effluent FP profiles. Fluorescence excitation-emission matrices (EEMs) were used with projection to latent structures (PLS) modeling to predict the FP and SProt concentrations in the effluent with average errors below 0.75 and 0.43 g L-1, respectively. Both models were able to capture the tendency of the data even when the accuracy of prediction was not so high. The combined approach of using 2D fluorescence spectroscopy and mathematical analysis seemed promising for real-time monitoring of the acidogenic fermentation of complex substrates.publishersversioninpres

A two-stage process for conversion of brewer’s spent grain into volatile fatty acids through acidogenic fermentation

This work is focused on the valorization of brewer’s spent grains (BSG) into volatile fatty acids (VFA) through acidogenic fermentation. VFAs are building blocks for several applications, such as bioplastics’ production. Using acid hydrolysis as pre-treatment, several batch assays were performed and the impact of organic load (OL) and pH on VFA production from BSG hydrolysate was assessed. Regardless of the condition, the produced acids were mainly butyric and acetic acids followed by propionic acid. The OL had a direct impact on the total organic acid concentration with higher concentrations at the highest OL (40 gCOD L-1). pH affected the concentration of individual organic acid, with the highest fermentation products (FP) diversity attained at pH 5.0 and OL of 40 gCOD L-1. To assess the potential application of organic acids for biopolymers (such as polyhydroxyalkanoates) production, the content in hydroxybutyrate (HB) and hydroxyvalerate (HV) monomers was estimated from the respective precursors produced at each pH and OL. The content in HV precursors increased with pH, with a maximum at pH 6.0 (ca. 16% C-mol basis). The acidogenic fermentation of BSG hydrolysate was also assessed in continuous operation, using an expanded granular sludge bed reactor (EGSB). It was shown that the BSG hydrolysate was successfully converted to VFAs without pH control, achieving higher productivities than in the batch operation mode.publishersversionpublishe

Valorization of brewery waste through Polyhydroxyalkanoates production supported by a metabolic specialized microbiome

Raw brewers’ spent grain (BSG), a by-product of beer production and produced at a large scale, presents a composition that has been shown to have potential as feedstock for several biological processes, such as polyhydroxyalkanoates (PHAs) production. Although the high interest in the PHA production from waste, the bioconversion of BSG into PHA using microbial mixed cultures (MMC) has not yet been explored. This study explored the feasibility to produce PHA from BSG through the enrichment of a mixed microbial culture in PHA-storing organisms. The increase in organic loading rate (OLR) was shown to have only a slight influence on the process performance, although a high selectivity in PHA-storing microorganisms accumulation was reached. The culture was enriched on various PHA-storing microorganisms, such as bacteria belonging to the Meganema, Carnobacterium, Leucobacter, and Paracocccus genera. The enrichment process led to specialization of the microbiome, but the high diversity in PHA-storing microorganisms could have contributed to the process stability and efficiency, allowing for achieving a maximum PHA content of 35.2 ± 5.5 wt.% (VSS basis) and a yield of 0.61 ± 0.09 CmmolPHA/CmmolVFA in the accumulation assays. Overall, the production of PHA from fermented BSG is a feasible process confirming the valorization potential of the feedstock through the production of added-value products.info:eu-repo/semantics/publishedVersio

Genetic diversity and population structure of Cynara cardunculus L. in southern Portugal

Cynara cardunculus L. is a cardoon species native to the Mediterranean region, which is composed of three botanical taxa, each having distinct biological characteristics. The aim of this study was to examine wild populations of C. cardunculus established in Portugal, in order to determine their genetic diversity, geographic distribution, and population structure. Based on SSR markers, 121 individuals of C. cardunculus from 17 wild populations of the Portuguese Alentejo region were identified and analysed. Ten SSRs were found to be efficient markers in the genetic diversity analysis. The total number of alleles ranged from 9 to 17 per locus. The expected and observed means in heterozygosity, by population analysed, were 0.591 and 0.577, respectively. The wild population exhibited a high level of genetic diversity at the species level. The highest proportion of genetic variation was identified within a geographic group, while variation was lower among groups. Geographic areas having highest genetic diversity were identified in Alvito, Herdade da Abo´boda, Herdade da Revilheira and Herdade de São Romão populations. Moreover, significant genetic differentiation existed between wild populations from North-Alentejo geographic locations (Arraiolos, E´ vora, Monte da Chamine´ ) and Centro Hortofrutı´cola, compared with other populations. This study reports genetic diversity among a representative number of wild populations and genotypes of C. cardunculus from Portugal. These results will provide valuable information towards future management of C. cardunculus germplasm.info:eu-repo/semantics/publishedVersio

COVID-19 in long-term care facilities in Brazil: serological survey in a post-outbreak setting

This cross-sectional seroepidemiological survey presents the seroprevalence of SARS-CoV-2 in a population living in 15 Long-Term Care Facilities (LTCFs), after two intra-institutional outbreaks of COVID-19 in the city of Botucatu, Sao Paulo State, Brazil. Residents were invited to participate in the serological survey performed in June and July 2020. Sociodemographic and clinical characterization of the participants as well as the LTCF profile were recorded. Blood samples were collected, processed and serum samples were tested using the rapid One Step COVID-19 immunochromatography test to detect IgM and IgG anti-SARS-CoV-2. Among 209 residents, the median of age was 81 years old, 135 (64.6%) were female and 171 (81.8%) self-referred as being white. An overall seroprevalence of 11.5% (95% CI: 7.5% – 16.6%) was found. The highest seroprevalences of 100% and 76.9% were observed in LTCFs that had experienced COVID-19 outbreaks. Most residents with positive immunochromatography tests (70.8%) referred previous contact with a confirmed COVID-19 case. Although there was a relatively low seroprevalence of COVID-19 in the total number of elderly people, this population is highly vulnerable and LTCFs are environments at higher risk for COVID-19 dissemination. A well-established test for COVID-19 policies, the adequate characterization of the level of interaction between residents and the healthcare provider team and the level of complexity of care are crucial to monitor and control the transmission of SARS-CoV-2 in these institutions

Tipificação de amostras aviárias patogênicas de Escherichia coli pela REP-PCR

In the present study the repetitive extragenic palindromic (REP) polymerase chain reaction (PCR) technique was used to establish the clonal variability of 49 avian Escherichia coli (APEC) strains isolated from different outbreak cases of septicemia (n=24), swollen head syndrome (n=14) and omphalitis (n=11). Thirty commensal strains isolated from poultry with no signs of these illnesses were used as control strains. The purified DNA of these strains produced electrophoretic profiles ranging from 0 to 15 bands with molecular sizes varying from 100 bp to 6.1 kb, allowing the grouping of the 79 strains into a dendrogram containing 49 REP-types. Although REP-PCR showed good discriminating power it was not able to group the strains either into specific pathogenic classes or to differentiate between pathogenic and non-pathogenic strains. On the contrary, we recently demonstrated that other techniques such as ERIC-PCR and isoenzyme profiles are appropriate to discriminate between commensal and APEC strains and also to group these strains into specific pathogenic classes. In conclusion, REP-PCR seems to be a technique neither efficient nor universal for APEC strains discrimination. However, the population clonal structure obtained with the use of REP-PCR must not be ignored particularly if one takes into account that the APEC pathogenic mechanisms are not completely understood yet.A técnica de REP (Repetitive extragenic palindrome)-PCR foi utilizada para avaliar a variabilidade genética de 49 amostras de Escherichia coli patogênicas para aves (APEC), isoladas de aves de corte (frangos) em diferentes surtos de septicemia (n=24), síndrome da cabeça inchada (n=14) e onfalite (n=11). Trinta amostras comensais, isoladas de frangos sem sinais de doença, foram utilizadas como controle. A análise do perfil eletroforético obtido por reação de REP-PCR utilizando DNA purificado das amostras evidenciou a amplificação de 0 a 15 bandas de DNA com pesos moleculares variando entre 100 pb e 6.1 Kb. A análise deste padrão permitiu a construção de um dendrograma demonstrando o agrupamento das 79 amostras em 49 perfis distintos. Embora a técnica de REP-PCR tenha apresentado grande poder discriminatório, as amostras patogênicas e não patogênicas não foram discriminadas entre si assim como não foi observado o agrupamento de amostras causadoras do mesmo tipo de doença. Por outro lado, demonstramos recentemente que outras técnicas tais como ERIC-PCR e a análise de isoenzimas foram eficientes quando utilizadas para esta mesma finalidade. Concluindo, REP-PCR parece não ser uma técnica eficiente e universal para discriminar entre amostras APEC. Porém, a estrutura clonal populacional obtida com o uso de REP-PCR não deve ser desprezada, particularmente se considerarmos que os mecanismos de patogenicidade de APEC ainda não são completamente conhecidos.6973Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

β-cyclodextrin/isopentyl caffeate inclusion complex: synthesis, characterization and antileishmanial activity

Isopentyl caffeate (ICaf) is a bioactive ester widely distributed in nature. Our patented work has shown promising results of this molecule against Leishmania. However, ICaf shows poor solubility, which limits its usage in clinical settings. In this work, we have proposed the development of an inclusion complex of ICaf in -cyclodextrin (-CD), with the aim to improve the drug solubility, and thus, its bioavailability. The inclusion complex (ICaf:-CD) was developed applying three distinct methods, i.e., physical mixture (PM), kneading (KN) or co-evaporation (CO) in different molar proportions (0.25:1, 1:1 and 2:1). Characterization of the complexes was carried out by thermal analysis, Fourier-transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM) and molecular docking. The ICaf:-CD complex in a molar ratio of 1:1 obtained by CO showed the best complexation and, therefore, was selected for further analysis. Solubility assay showed a marked improvement in the ICaf:-CD (CO, 1:1) solubility profile when compared to the pure ICaf compound. Cell proliferation assay using ICaf:-CD complex showed an IC50 of 3.8 and 2.7 µg/mL against L. amazonesis and L. chagasi promastigotes, respectively. These results demonstrate the great potential of the inclusion complex to improve the treatment options for visceral and cutaneous leishmaniases.This research was funded by Banco do Nordeste (grant FUNDECI/2016.0015), Coordenação Aperfeiçoamento de Pessoal de Nivel Superior (CAPES) and Fundação de Ámparo à Pesquisa do Estado de Sergipe (FAPITEC) (PROCESSO: 88887.159533/2017-00 extração, encapsulação e caracterização de bioativos para o interesse biotecnologico). Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq 301964/2019-0 Chamada 06/2019, and Chamada CNPq nº 01/2019) and from the Portuguese Science and Technology Foundation (FCT) project UIDB/04469/2020 (strategic fund).info:eu-repo/semantics/publishedVersio