Thermal hydrolysis pre-treatment has no positive influence on volatile fatty acids production from sewage sludge

The study compares the potential to produce volatile fatty acids (VFA) from sewage sludge, both raw and thermally pre-treated in two modes of operation. In batch mode, raw sludge at pH 8 obtained the highest maximum VFA yield (0.41 g COD-VFA/g CODfed) whereas pre-treated sludge achieved a lower value (0.27 g COD-VFA/g CODfed). The operation of 5-L continuous reactors showed that thermal hydrolysis pre-treatment (THP) did not have any significant influence on VFA yields, averaging 15.1 % g COD-VFA/g COD with raw sludge and 16.6 % g COD-VFA/g COD with pre-treated one. Microbial community analysis showed that phylum Firmicutes was predominant in both reactors and that the enzymatic profiles involved in VFA production were very similar regardless of the substrate fedThis work is part of the ECOVAL and CIGAT CIRCULAR projects, which are funded by Interreg Sudoe (SOE4/P1/E1104) and by the Xunta de Galicia and Viaqua (IN853C2022/03), respectively. Juan M. Lema belongs to the Galician competitive research group ED431C-2021/37 co-funded by Xunta de Galicia and ERDF (EU)S

Influence of metabolism and microbiology on organic micropollutants biotransformation in anoxic heterotrophic reactors

There is scarce information about the biotransformation of organic micropollutants (OMPs) under anoxic conditions. In this study, a heterotrophic denitrifying bioreactor was set up to study the fate of several OMPs from metabolic and microbiological points of view. Primary metabolic activity was increased by adding progressively higher nitrogen loading rates during the operation (from 0.075 to 0.4 g N-NO3- L−1 d−1), which resulted in an important shift in the microbial population from a specialized biomass to a more diverse community. Such a change provoked a significant increase in the removal efficiency of erythromycin (ERY), roxithromycin (ROX) and bisphenol-A (BPA), and some bacterial taxa, such as Rhodoplanes, were identified as possible indicators related to the biodegradation of these compounds. The increasing primary metabolic activity in the reactor did not enhance the OMP-specific removal rates, suggesting that the bacterial composition is more influential than cometabolismThis research was supported by the Spanish Research State Agency (AEI) through ANTARES (PID2019–110346RB-C21) project. M. Martinez-Quintela would also like to express his gratitude to the same agency for awarding a research scholarship (BES-2017–080503). All authors belong to the Galician Competitive Research Groups (GRC)_ ED431C-2021/37S

Study of the microbiota associated to Ruditapes decussatus and Ruditapes philippinarum clams by 16S rRNA metabarcoding, dilution to extinction, and culture-based techniques

The study of the microbiota associated to clams is important not only to know their sanitary status but also to prevent pathobiology events. The use of different microbiological techniques can help to obtain a better picture of the bacterial diversity of clams as well as to isolate new bacterial taxa. In this study, two clam species, Ruditapes decussatus and R. philippinarum, were analyzed in two locations of Galicia (northwest of Spain) in April and October, by combining classic culturing, dilution-to-extinction approach, and 16S rRNA gene target sequencing. 16S rRNA gene target sequencing revealed a great diversity within the clam samples, shedding light into the vast microbial communities associated to these bivalves. All samples were dominated by the same bacterial genera in the different periods, namely Mycoplasma, Vibrio, and Cutibacterium. The α-diversity in the samples obtained during the month of October was lower and showed the dominance of rare bacterial taxa, such as Methylobacterium or Psychrobacter. Dilution-to-extinction technique demonstrated its usefulness to culture rare bacterial taxa that were not found in clams under the classic culturing techniques, including Rahnella, Brachybacterium, Micrococcus, Jantinobacter, and Lelliottia. Altogether, our study provides valuable information on the microbiota associated to R. decussatus and R. philippinarum, demonstrating the high complexity and dynamics of these microbial populationsOpen Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature. This work was supported in part by grant AGL2013-42628-R and AGL2016-77539-R from the Ministerio de Economía y Competitividad (Spain)S

Sulfamethoxazole enhances specific enzymatic activities under aerobic heterotrophic conditions: a metaproteomic approach

The growing concern about antibiotic-resistant microorganisms has focused on the sludge from wastewater treatment plants (WWTPs) as a potential hotspot for their development and spread. To this end, it seems relevant to analyze the changes on the microbiota as a consequence of the antibiotics that wastewater may contain. This study aims at determining whether the presence of sulfamethoxazole (SMX), even in relatively low concentrations, modifies the microbial activities and the enzymatic expression of an activated sludge under aerobic heterotrophic conditions. For that purpose, we applied a metaproteomic approach in combination with genomic and transformation product analyses. SMX was biotransformed, and the metabolite 2,4(1H,3H)-pteridinedione-SMX (PtO-SMX) from the pterin-conjugation pathway was detected at all concentrations tested. Metaproteomics showed that SMX at 50–2000 μg/L slightly affected the microbial community structure, which was confirmed by DNA metabarcoding. Interestingly, an enhanced activity of the genus Corynebacterium and specifically of five enzymes involved in its central carbon metabolism was found at increased SMX concentrations. Our results suggest a role of Corynebacterium genus on SMX risks mitigation in our bioreactorsThis research was funded by the Spanish Government (Agencia Estatal de Investigación) through the ANTARES project (PID2019-110346RB-C21), a PhD Xunta de Galicia Grant (ED481A-2018/113, D.M.K.-V.) and a Juan de la Cierva-Formación postdoctoral grant (FJC2019-041664-I, A.T.-S.). The authors belong to the Galician Competitive Research Groups (GRC_ ED431C-2021/37)S

Revisiting the Taxonomy of the Genus Arcobacter: Getting Order From the Chaos

Since the description of the genus Arcobacter in 1991, a total of 27 species have been described, although some species have shown 16S rRNA similarities below 95%, which is the cut-off that usually separates species that belong to different genera. The objective of the present study was to reassess the taxonomy of the genus Arcobacter using information derived from the core genome (286 genes), a Multilocus Sequence Analysis (MLSA) with 13 housekeeping genes, as well as different genomic indexes like Average Nucleotide Identity (ANI), in silico DNA–DNA hybridization (isDDH), Average Amino-acid Identity (AAI), Percentage of Conserved Proteins (POCPs), and Relative Synonymous Codon Usage (RSCU). The study included a total of 39 strains that represent all the 27 species included in the genus Arcobacter together with 13 strains that are potentially new species, and the analysis of 57 genomes. The different phylogenetic analyses showed that the Arcobacter species grouped into four clusters. In addition, A. lekithochrous and the candidatus species ‘A. aquaticus’ appeared, as did A. nitrofigilis, the type species of the genus, in separate branches. Furthermore, the genomic indices ANI and isDDH not only confirmed that all the species were well-defined, but also the coherence of the clusters. The AAI and POCP values showed intra-cluster ranges above the respective cut-off values of 60% and 50% described for species belonging to the same genus. Phenotypic analysis showed that certain test combinations could allow the differentiation of the four clusters and the three orphan species established by the phylogenetic and genomic analyses. The origin of the strains showed that each of the clusters embraced species recovered from a common or related environment. The results obtained enable the division of the current genus Arcobacter in at least seven different genera, for which the names Arcobacter, Aliiarcobacter gen. nov., Pseudoarcobacter gen. nov., Haloarcobacter gen. nov., Malacobacter gen. nov., Poseidonibacter gen. nov., and Candidate ‘Arcomarinus’ gen. nov. are proposedThis work was supported in part by Grants JPIW2013-69095-C03-03 from the Ministerio de Economía y Competitividad (MINECO), AQUAVALENS of the Seventh Framework Program (FP7/2007-2013) grant agreement 311846 from the European Union and AGL2013-42628-R and AGL2016-77539-R (AEI/FEDER UE) from the Agencia Estatal de Investigación (Spain)S

The fate of SARS-COV-2 in WWTPS points out the sludge line as a suitable spot for detection of COVID- 19

SARS-CoV-2 genetic material is detectable in the faeces of a considerable part of COVID-19 cases and hence, in municipal wastewater. This fact was confirmed early during the spread of the COVID-19 pandemic and prompted several studies that proposed monitoring its incidence by wastewater. This paper studies the fate of SARS-CoV-2 genetic material in wastewater treatment plants using RT-qPCR with a two-fold goal: i) to check its presence in the water effluent and in the produced sludge and ii) based on the understanding of the virus particles fate, to identify the most suitable spots for detecting the incidence of COVID-19 and monitor its evolution. On the grounds of the affinity of enveloped virus towards biosolids, we hypothesized that the sludge line acts as a concentrator of SARS-CoV-2 genetic material. Sampling several spots in primary, secondary and sludge treatment at the Ourense (Spain) WWTP in 5 different days showed that, in effect, most of SARS-CoV-2 particles cannot be detected in the water effluent as they are retained by the sludge line. We identified the sludge thickener as a suitable spot for detecting SARS-CoV-2 particles thanks to its higher solids concentration (more virus particles) and longer residence time (less sensitive to dilution caused by precipitation). These findings could be useful to develop a suitable strategy for early warning of COVID-19 incidence based on WWTP monitoringSabela Balboa, Miguel Mauricio-Iglesias and Juan M. Lema belong the following programs co-funded by ERDF (EU): the CRETUS Strategic Partnership (ED431E 2018/01) and the Galician Competitive Research Group (ED431C2017/029)S

Estudio de la variabilidad intraespecífica del patógeno de almejas Vibrio tapetis

Vibrio tapetis, microorganismo que se aisló por primera vez en 1990, es el agente etiológico de la enfermedad del anillo marrón (BRD, del inglés Brown Ring Disease), uno de los principales factores limitantes del cultivo de la almeja japonesa (Venerupis philippinarum) en Europa