Autochthonous Microbial Communities Research Articles

ПОЧВЕННЫЕ БАКТЕРИИ ИЗ АВТОХТОННЫХ МИКРОБНЫХ СООБЩЕСТВ ЦЕНТРАЛЬНОЙ СИБИРИ В БИОЛОГИЧЕСКОЙ ЗАЩИТЕ ПШЕНИЦЫ ОТ ALTERNARIA TENUISSIMA

The most powerful soil bacteria from autochthonous microbial communities of Central Siberia against phytopathogenic fungi Alternaria tenuissima were selected. Isolation from soil samples and cultivation of bacterial isolates were performed on the artificial nutrient medium for bacteria PD-agar. Antagonistic activity was measured on GRM Saburo. In this case, Alternaria tenuissima was sown in a dish on GRM Saburo, and the selected bacterial strains were sown around the micromycete. As a result of joint growth of the test culture and soil bacteria, the most powerful antagonists in the suppression zone of Alternaria tenuissima were identified. The most powerful bacteria-antagonists in reducing the intensity and prevalence of root rot were identified by the method of artificial infection of Alternaria tenuissima. Before infection with micromycete, bacterization of spring wheat seeds Novosibirskaya-15 with the studied bacterial isolates was carried out. Mathematical processing of the obtained results was fulfilled by one-way variance analysis and two-sample F-test for variance. The results of the study on the use of soil autochthonous antagonist microorganisms in biological protection of spring wheat from the causative agents of root rot Alternaria tenuissima showed that the studied bacterial isolates statistically significantly (p < 0.01) reduced the intensity and prevalence of this disease. The maximum effect in reducing the intensity and prevalence of the disease was shown by isolates B2 and B4 (according to preliminary identification of Bacillus sp.). Also, the studied bacterial strains showed a positive effect on the length of sprouts. At the same time, isolate B2 showed a statistically significant (p < 0.001) effect on the length of spring wheat sprouts. The stimulating effect was manifested in an increase in the length of sprouts compared to the control. Therefore, this strain can be recommended not only for protection against Alternaria tenuissima, but also for stimulating wheat growth.

Engineering multi-degrading bacterial communities to bioremediate soils contaminated with pesticides residues

Parallel to the important use of pesticides in conventional agriculture there is a growing interest for green technologies to clear contaminated soil from pesticides and their degradation products. Bioaugmentation i. e. the inoculation of degrading micro-organisms in polluted soil, is a promising method still in needs of further developments. Specifically, improvements in the understanding of how degrading microorganisms must overcome abiotic filters and interact with the autochthonous microbial communities are needed in order to efficiently design bioremediation strategies. Here we designed a protocol aiming at studying the degradation of two herbicides, glyphosate (GLY) and isoproturon (IPU), via experimental modifications of two source bacterial communities. We used statistical methods stemming from genomic prediction to link community composition to herbicides degradation potentials. Our approach proved to be efficient with correlation estimates over 0.8 - between model predictions and measured pesticide degradation values. Multi-degrading bacterial communities were obtained by coalescing bacterial communities with high GLY or IPU degradation ability based on their community-level properties. Finally, we evaluated the efficiency of constructed multi-degrading communities to remove pesticide contamination in a different soil. While results are less clear in the case of GLY, we showed an efficient transfer of degrading capacities towards the receiving soil even at relatively low inoculation levels in the case of IPU. Altogether, we developed an innovative protocol for building multi-degrading simplified bacterial communities with the help of genomic prediction tools and coalescence, and proved their efficiency in a contaminated soil.

ПОЧВЕННЫЕ БАКТЕРИИ ИЗ АВТОХТОННЫХ МИКРОБНЫХ СООБЩЕСТВ ЦЕНТРАЛЬНОЙ СИБИРИ В БИОЛОГИЧЕСКОЙ ЗАЩИТЕ ПШЕНИЦЫ ОТ ALTERNARIA TENUISSIMA

The aim of the study is to find the strongest soil microorganisms from autochthonous microbial communities of Central Siberia against phytopathogenic micromycetes Alternaria tenuissima. The object of the study is autochthonous soil bacteria isolated from the soils of the Krasnoyarsk forest-steppe in the Sukhobuzimsky District. Soil samples were taken from under spring wheat Novosibirskaya 15 and from under perennial grasses. The soil cover is represented by ordinary leached chernozem. Ammonium nitrate was used (34.7 kg/ha active ingredient). The climatic conditions are moderately dry and continental. The studies were carried out in 2022–2023. Antagonistic activity was measured on the GRM Saburo. With a general growth of the test culture and soil bacteria, the strongest antagonists were found in the suppression zone of Alternaria tenuissima. The identification of the most powerful bacterial antagonists in reducing the intensity and prevalence of root rot was carried out using the method of artificial infection with Alternaria tenuissima. Before infection with micromycete, bacterization of spring wheat seeds Novosibirskaya 15 with the studied bacterial isolates was carried out. Mathematical processing of the obtained results was carried out by one-way variance analysis and two-sample F-test for variance. The results of the study on the use of soil autochthonous antagonist microorganisms in biological protection of spring wheat from the causative agents of root rot Alternaria tenuissima showed that the studied bacterial isolates statistically significantly (p < 0.01) reduced the intensity and prevalence of this disease. The maxi-mum effect in reducing the intensity and prevalence of the disease was shown by isolates B2 and B4. Also, the studied bacterial strains showed a significant effect (p < 0.001) on the length of sprouts. The greatest increase in length in spring wheat sprouts was in the experimental variant with strain B2. This strain can be recommended not only for protection against Alternaria tenuissima, but also for stimuli-ting wheat growth.

Synthesis of an autochthonous microbial community by analyzing the core microorganisms responsible for the critical flavor of bran vinegar

Traditional bran vinegar brewing unfolds through natural fermentation, a process driven by spontaneous microbial activity. The unique metabolic activities of various microorganisms lead to distinct flavors and qualities in each batch of vinegar, making it challenging to consistently achieve the desired characteristic flavor compounds. Therefore, identifying the critical microbial species responsible for flavor production and designing starter cultures with improved fermentation efficiency and characteristic flavors are effective methods to address this discrepancy. In this study, 11 core functional microbial species affecting the fermentation flavor of Sichuan shai vinegar (Cupei were placed outside solarization and night-dew for more than one year, and vinegar was the liquid leached from Cupei) (SSV), were revealed by combining PacBio full-length diversity sequencing based on previous metagenomics. The effects of environmental factors and microbial interactions on the growth of 11 microorganisms during fermentation were verified using fermentation experiments. Ultimately, the microbial community was strategically synthesized using a 'top-down' approach, successfully replicating the distinctive flavor profile of Sichuan shai vinegar (SSV). The results showed that the interaction between microorganisms and environmental factors affected microorganism growth. Compared with traditional fermentation, the synthetic microbial community’s vinegar-fermented grains (Cupei) can reproduce the key flavor of SSV and is conducive to the production of amino acids. In this study, the key flavor of SSV was reproduced through rational design of the synthetic microbial community. This achievement holds profound significance for the broader application of microbiome assembly strategies in the realm of fermented foods.

Isolation, identification, and stability of sourdough microbiota from spontaneously fermented Norwegian legumes

Fermentation has recently been rediscovered as an attractive technique to process legumes, as it can improve the nutritional quality and value of the end product.This study investigated the dynamics and stability of the microbial communities in spontaneously fermented sourdoughs made from flours of two cultivars of faba beans and two cultivars of peas. Sourdoughs were established by the backslopping technique, and the microbial development at 22 °C and 30 °C was followed by culture dependent and culture independent methods. The utilization of substrates and formation of metabolites were also determined by high-performance liquid chromatography.A stable pH was reached in all the sourdoughs after 11–15 days of daily backslopping. Lactic acid bacteria and yeast from pH stable sourdoughs were isolated, characterized and identified. The fermentation temperature influenced the development of the microbial community and the substrate utilization during spontaneous fermentation. In the 30 °C fermentations, one species dominated (Lactiplantibacillus plantarum/pentosus), a lower pH was achieved, and the available substrates were more extensively converted. The 22 °C fermentation resulted in a more diverse microbial community (Lactiplantibacillus, Leuconostoc, Pediococcus), a higher pH, and more residual substrates were available after fermentation. Yeasts were only detected in one of the pea sourdoughs fermented at 30 °C, with Saccharomyces cerevisiae being the dominant species. Nearly all sourdoughs were depleted of maltose after 24 h fermentation cycles, and higher levels of lactic and acetic acid were detected in 30 °C fermen-tations.This research adds to our understanding of the autochthonous microbial community present in faba beans and peas as well as their natural capacity to establish themselves and ferment legume flours. These findings enhance the possibilities of utilizing and improving plant based protein sources.

Site-specific response of sediment microbial community to supplementation of polyhydroxyalkanoates as biostimulants for PCB reductive dechlorination

The use of biodegradable plastics is constantly raising, increasing the likeliness for these polymers to end up in the environment. Environmental applications foreseeing the intentional release of biodegradable plastics have been also recently proposed, e.g., for polyhydroxyalkanoates (PHAs) acting as slow hydrogen releasing compounds to stimulate microbial reductive dehalogenation processes. However, the effects of their release into the environment on the ecosystems still need to be thoroughly explored. In this work, the use of PHAs to enhance the microbial reductive dechlorination of polychlorobiphenyls (PCBs) and their impact on the metabolic and compositional features of the resident microbial community have been investigated in laboratory microcosms of a polluted marine sediment from Mar Piccolo (Taranto, Italy), and compared with recent findings on a different contaminated marine sediment from Pialassa della Baiona (Ravenna, Italy). A decreased biostimulation efficiency of PHAs on PCBs reductive dechlorination was observed in the sediment from Mar Piccolo, with respect to the sediment from Pialassa della Baiona, suggesting that the sediments' physical-chemical characteristics and/or the biodiversity and composition of its microbial community might play a key role in determining the outcome of this biostimulation strategy. Regardless of the sediment origin, PHAs were found to have a specific and pervasive effect on the sediment microbial community, reducing its biodiversity, defining a newly arranged microbial core of primary degraders and consequently affecting, in a site-specific way, the abundance of subdominant bacteria, possibly cross-feeders. Such potential to dramatically change the structure of autochthonous microbial communities should be carefully considered, since it might have secondary effects, e.g., on the natural biogeochemical cycles.

Microbial Ecology of Pecorino Siciliano PDO Cheese Production Systems

Pecorino Siciliano PDO is a semi-hard cheese that is produced in wooden vats using raw sheep’s milk and its associated autochthonous microbial community. In the present study, we evaluated the microbial ecology of the milk, curd and whey from five Pecorino Siciliano PDO-producing farms in Sicily using a combination of metagenomic and microbiological approaches. We present an overview of the species and strain-level diversity of dairy lactococcal and streptococcal isolates using established genotyping tools and compare the lactic acid bacterial populations present in samples from these farms. Whole genome sequences of representative isolates of Lactococcus spp. and Streptococcus thermophilus were elucidated and the genetic diversity of the strains was established through analysis of predicted phage-resistance systems and prophage-associated regions. The analysis revealed farm-specific dairy lactococcal and streptococcal isolates that possess diverse genotypic features including newly described phage-resistance systems.

Regulation of the autochthonous microbial community in excess sludge for the bioconversion of carbon dioxide to acetate without exogenic hydrogen

The autochthonous microbial community from excess sludge was regulated for enhanced conversion of CO2 to acetate without exogenic H2. It was interesting that the acetate-fed system exhibited a surprising performance to regulate the microbial community for a high acetate yield and selectivity. As a result, some hydrogen-producing bacteria (e.g., Proteiniborus) and acetogenic bacteria with the ability of CO2 reduction were enriched by acetate feeding, 2-bromoethanesulfonate (BES) addition and CO2 stress. When the selected microbial community was applied to convert CO2, the accumulation of acetate was positively correlated to the concentration of yeast extract. Finally, the acetate yield reached up to 67.24 mM with a high product selectivity of 84 % in the presence of yeast extract (2 g/L) and sufficient CO2 in semi-continuous culture for 10 days. This work should help get new insights into the regulation of microbial community for the efficient acetate production from CO2.

A deep continental aquifer downhole sampler for microbiological studies.

To be effective, microbiological studies of deep aquifers must be free from surface microbial contaminants and from infrastructures allowing access to formation water (wellheads, well completions). Many microbiological studies are based on water samples obtained after rinsing a well without guaranteeing the absence of contaminants from the biofilm development in the pipes. The protocol described in this paper presents the adaptation, preparation, sterilization and deployment of a commercial downhole sampler (PDSshort, Leutert, Germany) for the microbiological studying of deep aquifers. The ATEX sampler (i.e., explosive atmospheres) can be deployed for geological gas storage (methane, hydrogen). To validate our procedure and confirm the need to use such a device, cell counting and bacterial taxonomic diversity based on high-throughput sequencing for different water samples taken at the wellhead or at depth using the downhole sampler were compared and discussed. The results show that even after extensive rinsing (7 bore volumes), the water collected at the wellhead was not free of microbial contaminants, as shown by beta-diversity analysis. The downhole sampler procedure was the only way to ensure the purity of the formation water samples from the microbiological point of view. In addition, the downhole sampler allowed the formation water and the autochthonous microbial community to be maintained at in situ pressure for laboratory analysis. The prevention of the contamination of the sample and the preservation of its representativeness are key to guaranteeing the best interpretations and understanding of the functioning of the deep biosphere.

Correlation between autochthonous microbial communities and flavor profiles during the fermentation of mustard green paocai (Brassica juncea Coss.), a typical industrial-scaled salted fermented vegetable

Mustard green paocai is a salted vegetable product that is fermented with low salt in the initial stage and preserved with high salt in the middle stage. To reveal the key flavor-producing potential of core bacterial and fungal communities during the 365-day fermentation of industrial-scaled paocai. In this study, microbiome and molecular sensory science were used to analyze potential relationships between microbial diversity and key flavor compounds. The results showed that bacteria were more diverse than fungi. High salinity improved the flavor of industrial paocai and simplified the microbial community. A total of 74 bacterial and yeast strains were isolated and identified throughout the fermentation process. Lactiplantibacillus, Halanaerobium, Halomonas, Pseudomonas, and Enterobacteriaceae were the dominant bacterial genera, and Debaryomyces hansenii was the core fungal genera in paocai. 4-Ethylphenol, anethole, butanoic acid, 4-ethylguaiacol, and 3-methyl-1-butanol were identified as key flavor compounds in industrial paocai with odor activity values greater than 1. Spearman correlation analysis indicated that bacteria have a greater impact on the flavor of paocai than fungi. Halanaerobium and Halomonas were correlated with flavor and spoilage in paocai. This study provides guidance for the production of desirable flavors by multi-bacteria synergistic fermentation of industrial paocai.

Evaluating the aerobic xylene-degrading potential of the intrinsic microbial community of a legacy BTEX-contaminated aquifer by enrichment culturing coupled with multi-omics analysis: uncovering the role of Hydrogenophaga strains in xylene degradation

To develop effective bioremediation strategies, it is always important to explore autochthonous microbial community diversity using substrate-specific enrichment. The primary objective of this present study was to reveal the diversity of aerobic xylene-degrading bacteria at a legacy BTEX-contaminated site where xylene is the predominant contaminant, as well as to identify potential indigenous strains that could effectively degrade xylenes, in order to better understand the underlying facts about xylene degradation using a multi-omics approach. Henceforward, parallel aerobic microcosms were set up using different xylene isomers as the sole carbon source to investigate evolved bacterial communities using both culture-dependent and independent methods. Research outcome showed that the autochthonous community of this legacy BTEX-contaminated site has the capability to remove all of the xylene isomers from the environment aerobically employing different bacterial groups for different xylene isomers. Interestingly, polyphasic analysis of the enrichments disclose that the community composition of the o-xylene-degrading enrichment community was utterly distinct from that of the m- and p-xylene-degrading enrichments. Although in each of the enrichments Pseudomonas and Acidovorax were the dominant genera, in the case of o-xylene-degrading enrichment Rhodococcus was the main player. Among the isolates, two Hydogenophaga strains, belonging to the same genomic species, were obtained from p-xylene-degrading enrichment, substantially able to degrade aromatic hydrocarbons including xylene isomers aerobically. Comparative whole-genome analysis of the strains revealed different genomic adaptations to aromatic hydrocarbon degradation, providing an explanation on their different xylene isomer-degrading abilities.

Microbiome-based therapeutics: Opportunity and challenges.

The autochthonous microbial communities comprising symbionts, commensals, and opportunistic pathogens living throughout the human body profoundly contribute to health by reducing disease susceptibility and maturing host immunity. The community compositions and functional repertoires of microbiomes present in the different body habitats are dynamic. The structural and functional balance of the human microbiome could be modulated by environmental factors, lifestyle, and host genetics. Several functions of the microbial community directly or indirectly modulate host cellular signaling pathways that are associated with energy assimilation, sensing and responding to environmental signals through the neuroendocrine pathways, and resistance against colonization of allochthonous microbiota with disease-causing potential. Both culture-dependent and independent characterizations of microbial community compositions and their functional attributes help us to recognize the importance of microbial diversity in individual's health and identify the microbes and their metabolites associated with health and diseases. Such an in-depth understanding of the human microbiome created avenues of microbiome-based translation research, which led to the discovery and development of large numbers of medical therapies. In this chapter, we discuss the current success of microbiome-based therapies for infectious and metabolic diseases, and the major bottleneck and challenges of translational research with the community of symbiotic microorganisms.

Enhanced pilot bioremediation of oily sludge from petroleum refinery disposal under hot-summer Mediterranean climate

Large pilot scale bioremediation approaches were implemented for the treatments of oily sludge (OS) characterised by alkaline pH (pH > 9), high concentration of metals (3% dry weight) and high total petroleum hydrocarbons content (TPH) rangingbetween 22,000 and 67,300 mg kg −1 from a Tunisian petroleum refinery. The treatments included bioaugmentation and biostimulation approaches with autochthonous isolated bacterial strains and consortia. Chemical, microbial, and ecotoxicological analyses were performed over a period of 180 days incubation. The bioremediation treatments favoured the development of Proteobacteria , Firmicutes and Bacteroidetes following an ecological succession of specialist bacterial groups, first associated to hydrocarbon degradation (e.g. Marinobacter and Alcanivorax ) that resulted in a greater extent of TPH-degradation (up to 80%), and the selection of metal resistant bacteria including Hyphomonas , Phaeobacter , and Desulfuromusa . The best performances were obtained when bioaugmentation and biostimulation were combined. Over 90% of the TPH initial concentration was degraded over 180 days, which was accompanied with a 3-fold reduction of ecotoxicity. Our study demonstrates the efficacy of large pilot scale bioremediation of highly contaminated oily sludge, providing the evidence that the management of autochthonous microbial communities is of paramount importance for the success of the bioremediation process. • Effective oil sludge bioremediation by managing bacterial community. • Optimal treatment combining bioaugmentation and biostimulation. • Bacterial community plasticity evidenced through ecological succession.

Degradation of organometallic pollutants of distillery wastewater by autochthonous bacterial community in biostimulation and bioaugmentation process

This study aimed to detoxify and degrade the organometallic pollutants from distillery wastewater by using an autochthonous microbial community via biostimulation and bioaugmentation process. Results revealed that the wastewater contained high concentrations of the metals i.e. Fe-2403; Zn-210.15; Cr- 22.825; Cu-73.62; Mg-27.30; Ni-14.425; and Pb-17.33 (mg L-1). The biostimulation and bioaugmentation process resulted from a substantial reduction (50–70%) in the pollution load. Scanning electron microscopy analysis showed bacterial community and their relationship with complex organometallic pollutants during the chemical reactions. The major identified organic pollutants in the control (untreated) samples were acetic acid, Oxo-,trimethylsilyl ester [CAS], Hydrocinnamic acid, p-[Trimethylsiloxy]-trimethylsilyl ester and tetradecanoic acid, trimethylsilyl ester [CAS] while some new metabolic products were generated as a by-product in bioaugmentation process. Therefore, the study showed that biostimulation and bioaugmentation were successful bioremediation strategies for the detoxification of distillery wastewater and restoration of organometallic polluted sites.

The Effects of Nisin-Producing Lactococcus lactis Strain Used as Probiotic on Gilthead Sea Bream (Sparus aurata) Growth, Gut Microbiota, and Transcriptional Response

The present research tested the effects of dietary nisin-producingLactococcus lactison growth performance, feed utilization, intestinal morphology, transcriptional response, and microbiota in gilthead sea bream (Sparus aurata). A feeding trial was conducted with fish weighting 70–90 g. Fish were tagged with passive, integrated transponders and distributed in nine 500 L tanks with 40 fish each. Fish were fed for 12 weeks with either a control (diet A) or experimental diets (diets B and C) in triplicate (3 tanks/diet). Extruded pellets of diets B and C were supplemented with a low (2 × 109CFU/kg) and a high (5 × 109CFU/kg) dose of probiotic, respectively. No significant differences were found between groups for the feed conversion ratio or specific growth rates. However, the final body weight of fish fed diet C was significantly higher than the control group with intermediate values for fish fed diet B. Histological analysis conducted using a semi-quantitative scoring system showed that probiotic did not alter the morphology of the intestine and did not trigger inflammation. With regard to the transcriptomic response, a customized PCR array layout was designed to simultaneously profile a panel of 44 selected genes. Significant differences in the expression of key genes involved in innate and acquired immunity were detected between fish fed probiotic and control diets. To analyze the microbiota associated to the feeds and the gut autochthonous microbial communities, we used the Illumina MiSeq platform for sequencing the 16S rRNA gene and a metagenomics pipeline based on VSEARCH and RDP databases. The analysis of gut microbiota revealed a lack of colonization of the probiotic in the host’s intestinal mucosa. However, probiotic did modulate the fish gut microbiota, confirming that colonization is not always necessary to induce host modification. In fact, diets B and C were enriched with Actinomycetales, as compared to diet A, which instead showed a higher percentage ofPseudomonas,Sphyngomonas, andLactobacillusgenera. These results were confirmed by the clear separation of gut bacterial community of fish fed with the probiotic from the bacterial community of control fish group in the beta-diversity and PLS-DA (supervised partial least-squares discriminant analysis) analyses.

Allium cepa L. Inoculation with a Consortium of Plant Growth-Promoting Bacteria: Effects on Plants, Soil, and the Autochthonous Microbial Community.

The present work was aimed at investigating the effects of a four bacterial strain consortium—Azospirillum brasilense, Gluconacetobacter diazotrophicus, Herbaspirillum seropedicae, and Burkholderia ambifaria—on Allium cepa L. and on soil health. The bacterial consortium was inoculated on seeds of two different onion varieties; inoculated and Control seeds (treated with autoclaved inoculum) were sown in an open-field and followed until harvest. Plant growth development parameters, as well as soil physico–chemical and molecular profiles (DNA extraction and 16S community sequencing on the Mi-Seq Illumina platform), were investigated. The results showed a positive influence of bacterial application on plant growth, with increased plant height (+18%), total chlorophylls (+42%), crop yields (+13%), and bulb dry matter (+3%) with respect to the Control. The differences between Control and treatments were also underlined in the bulb extracts in terms of total phenolic contents (+25%) and antioxidant activities (+20%). Soil fertility and microbial community structure and diversity were also positively affected by the bacterial inoculum. At harvest, the soil with the presence of the bacterial consortium showed an increase in total organic carbon, organic matter, and available phosphorus, as well as higher concentrations of nutrients than the Control. The ecological indexes calculated from the molecular profiles showed that community diversity was positively affected by the bacterial treatment. The present work showed the effective use of plant growth-promoting bacteria as a valid fertilization strategy to improve yield in productive landscapes whilst safeguarding soil biodiversity.

Relationship between aquifer biofilms and unattached microbial indicators of urban groundwater contamination.

Aquifers, springs and other groundwater-dependent ecosystems are threatened by urban land use, which causes water quality deterioration through nutrient loading, sewage infiltration, groundwater extraction and, along coasts, seawater intrusion. The presence of certain microbes in groundwater can indicate that an aquifer is anthropogenically contaminated. Interpretations made from observations of indicator microbes in groundwater are limited because the relationship between the presumably allochthonous indicator microbes and relevant autochthonous microbial communities has not been characterized. This study addressed whether autochthonous aquifer biofilms can influence the presence of presumed microbial indicators in groundwater, and simultaneously used microbial indicators to trace sources of urban contamination at a karst spring of conservation concern. These questions were approached using a 17-month time series analysis of attached biofilm and adjacent unattached bacteria in the submerged karst aquifer conduit associated with this spring. Environmental 16S rRNA gene sequencing was performed to characterize these communities, and community structure data were contextualized with groundwater geochemical and hydrogeological measurements. Linear regression models were developed to explain the relative abundance patterns of indicator microbes and other unattached microbes at this site. The results of this study suggest that dominant aquifer biofilms do not influence the presence of unattached microbial taxa that are presumed to be indicators of groundwater contamination, and generated new information about the origin of coliform bacteria at the study site. These results build confidence in the use of microbial indicators in groundwater-dependent ecosystem conservation strategies and inform future management plans for urban aquifers and springs worldwide.

Ecological impact of organochlorine pesticides consortium on autochthonous microbial community in agricultural soil

Organochlorine pesticides (OCPs) used in agricultural practices are of global concern due to their toxicological hazards on biomes of the impacted soil. Geochemistry and microbiome of OCPs-impacted (OW) soil was determined and compared with those of pristine (L1) soils. Microbiome of OW was based on sequencing total 16S rRNA genes of prokaryotes and Internal Transcribed Spacer (ITS2) regions between 5.8S and 28S rRNA genes of eukaryotes using Illumina MiSeq platform for bacterial and fungal communities, respectively. Geochemical properties of OW were assessed for ecological risks of OCPs on biota via risk quotient (RQ) and maximum cumulative ratio (MCR). It was established OW was polluted with 15 OCPs, along with consequential nitrate and phosphorous deficiencies. Ten of the 15 OCPs exerted severe ecological risk (RQ > 1: 4–992), of which endosulfan contributed 76% of the ecotoxicity (MCR = 1.3) on OW. The key players in OW were observed to be Enterobacteriaceae and Mortierellaceae represented by Escherichia and Mortierella taxa, respectively. Low abundance of Nitrospirae species and extinction of Glomeromycota in OW connoted serious toxicological consequences of the OCPs. Taxon XOR (Taxon Exclusive Or) analysis revealed 38,212 and 63,474 counts of bacterial and fungal species, respectively, were exclusively found in the impacted OW and possibly contributed to natural attenuation of the OCPs in the impacted agricultural soil. Conversely, 61,005 (bacteria) and 33,397 (fungi) species counts that were missing in OCPs-impacted OW, but present in pristine L1, opined the species as bio-indicators of OCPs ecotoxicity in agricultural soils. While the species tagged as bio-indicators would be valuable in monitoring OCPs pollution, those suggested to be players in self-recovery process will be invaluable to designing bioremediation strategies for OCPs-impacted agricultural soil.

Impact of treated sewage effluent on the bacterial community composition in an intermittent mediterranean stream

Water quality monitoring is essential to safeguard human and environmental health. The advent of next-generation sequencing techniques in recent years, which allow a more in-depth study of environmental microbial communities in the environment, could broaden the perspective of water quality monitoring to include impact of faecal pollution bacteria on ecosystem. In this study, 16 S rRNA amplicon sequencing was used to evaluate the impact of wastewater treatment plant (WWTP) effluent on autochthonous microbial communities of a temporary Mediterranean stream characterized by high flow seasonality (from 0.02 m3/s in winter to 0.006 m3/s in summer).Seven sampling campaigns were performed under different temperatures and streamflow conditions (winter and summer). Water samples were collected upstream (Upper) of the WWTP, the secondary effluent (EF) discharge and 75 m (P75) and 1000 m (P1000) downstream of the WWTP.A total of 5,593,724 sequences were obtained, giving rise to 20,650 amplicon sequence variants (ASV), which were further analysed and classified into phylum, class, family and genus. Each sample presented different distribution and abundance of taxa. Although taxon distribution and abundance differed in each sample, the microbial community structure of P75 resembled that of EF samples, and Upper and P1000 samples mostly clustered together. Alpha diversity showed the highest values for Upper and P1000 samples and presented seasonal differences, being higher in winter conditions of high streamflow and low temperature.Our results suggest the microbial ecology re-establishment, since autochthonous bacterial communities were able to recover from the impact of the WWTP effluent in 1 km. Alpha diversity results indicates a possible influence of environmental factors on the bacterial community structure. This study shows the potential of next-generation sequencing techniques as useful tools in water quality monitoring and management within the climate change scenario.

Co-presence of the anionic surfactant sodium lauryl ether sulphate and the pesticide chlorpyrifos and effects on a natural soil microbial community.

There is a growing concern about the simultaneous presence in the environment of different kinds of pollutants, because of the possible synergic or additive effects of chemical mixtures on ecosystems. Chlorpyrifos (CPF) is an organophosphate insecticide extensively used in agricultural practices. The anionic surfactant sodium lauryl ether sulphate (SLES) is the main component of several commercial products, including foaming agents used in underground mechanised excavation. Both compounds are produced and sold in high amounts worldwide and can be found in the environment as soil contaminants. The persistence of SLES and CPF in agricultural soils and their possible effects on the natural microbial community was evaluated in microcosms. The experimental set consisted of soil samples containing the autochthonous microbial community and treated with only SLES (70mg/kg), only CPF (2mg/kg) or with a mix of both compounds. Control microcosms (without the contaminants) were also performed. Soil samples were collected over the experimental period (0, 7, 14, 21 and 28days) and analysed for CPF, SLES and the main metabolite of CPF (3, 5, 6-trichloropyridinol, TCP). The half-life time (DT50) of each parent compound was estimated in all experimental conditions. At the same time, the abundance, activity and structure of the microbial community were also evaluated. The results showed that the co-presence of SLES and CPF did not substantially affect their persistence in soil (DT50 of 11 and 9days with co-presence and 13 and 10days, respectively, when alone); however, in the presence of SLES, a higher amount of the metabolite TCP was found. Interestingly, some differences were found in the bacterial community structure, abundance and activity among the various conditions.