Mycorrhizal fungi and phosphate fertilization in the production of Euterpe edulis seedlings

The present study evaluated the effect of arbuscular mycorrhizal fungi (AMF) inoculation on growth and nutrition of Euterpe edulis seedlings, supplemented or not with phosphate fertilization. The experiment was conducted in a greenhouse. The randomized block design, consisted of a 3x2 factorial arrangement and 4 replicates, with two phosphorus doses (0 and 50 mg dm-3 of soil), two microbiological treatments (Rhizophagus clarus; laroideoglomus etunicatum; R. clarus + C. etunicatum) and control (without fungus). Sowing and inoculation occurred concurrently in 2 kg plastic bags. Height, collar diameter, leaf area, dry shoot mass, macronutrient content and mycorrhizal colonization percentage were determined after 226 days. Regarding mycorrhizal colonization percentage, R. clarus resulted significantly beneficial for the production of E. edulis seedlings. In the absence of phosphate fertilization, R. clarus and mixed inoculum increased all biometric variables and macronutrient contents in seedlings. Therefore, it is concluded that AMF inoculations provide beneficial effects for growth and nutrition of E. edulis seedlings, resulting in more vigorous plants at a low-cost strategy. Highlights: Rhizophagus clarus resulted significantly beneficial for the production of E. edulis seedlings and in the absence of phosphate fertilization, and mixed inoculum ( clarus + Claroideoglomus etunicatum) increased all biometric variables and macronutrient contents in seedlings. The AMF inoculations provide beneficial effects for growth and nutrition of E. edulis seedlings, resulting in more vigorous plants at a low-cost strategy. The AMF, a biological agent of mutualistic associations with plants, constitutes an innovative approach to sustainable agriculture, contributing to increase plant survival rate, a key-factor for the successful reintroduction and conservation of E. edulis, as well as for its commercial exploitation.

Assessing Antibiotics Biodegradation and Effects at Sub-inhibitory Concentrations by Quantitative Microbial Community Deconvolution

Antibiotics in the environment cause widespread concern as a result of their potent inhibitory action on microbial growth and their role in potentially creating selective conditions for proliferation of antibiotic resistant bacteria. Comprising a carbon skeleton, antibiotics should be amenable to microbial biodegradation, but this is still largely uncharted territory because of their simultaneous strong toxicity. In this study, we estimated potential antibiotics degradation by and effects on mixed microbial communities at concentrations sufficiently high to allow sensitive detection of biomass growth, but simultaneously, low enough to mitigate their toxic action. We used three different mixed inoculum sources freshly derived from freshwater, activated sludge or soil, and tested a series of 15 antibiotics from different classes at 1 mg C-carbon l−1 dosage. Consistent community growth was observed for freshwater and activated sludge with ampicillin, erythromycin and chloramphenicol, and with sulfomethoxazole for activated sludge, which was accompanied by parent compound disappearance. Community growth could be attributed to a few subclasses of recognized cell types by using supervised machine-learning-based classifiers. Most other tested antibiotics resulted in inhibition of community growth on background assimilable organic carbon, concomitant with altered composition of the resulting communities. We conclude that growth-linked biodegradation of antibiotics at low concentrations may be present among typical environmental microbiota, but for a selected subset only, whereas for the majority of antibiotics negative effects prevail without any sign of productive growth.

Rational Selection of Mixed Yeasts Starters for Agave Must Fermentation

Tequila and mezcal are both traditional Mexican liquors that are produced from cooked Agave spp. must fermentation and usually rely on spontaneous or pure Saccharomyces cerevisiae strain inoculation. In order to contribute to the rational selection of yeast starters for tequila and mezcal productions, we tested a collection of 25 yeasts originally isolated from mezcal musts, spanning 10 different yeast species. These strains were first characterized in a semi synthetic medium (labeled as M2, having 90 g/L fructose and 10 g/L glucose of initial hexoses) at 48 h of culture, observing a differential pattern in the consumption of sugars and productivity. Selected Saccharomyces strains left around 10 g/L of fructose and showed higher fermentation performance. However, some non-Saccharomyces strains, specifically from Torulospora (Td), Kluyveromyces (Km), and Zygosaccharomyces (Zb) genera, consumed almost all the sugar (i.e., Km1Y9 with <5 g/L) and had a high productivity of ethanol. In general, all Saccharomyces strains presented a high production of ethyl-butyrate, ethyl-decanoate, and ethyl-hexanoate with peaks of 10, 38, and 3 μg/L, respectively. In addition, some Kluyveromyces and Torulospora strains showed a high production of phenyl ethyl acetate (i.e., Km1D5 with up to 1400 μg/L); isoamyl acetate (i.e., Km1D5 and Td1AN2 with more than 300 μg/L), and hexyl acetate (i.e., Td1AN2 with 0.3 μg/L). Representative strains of the most productive genera (Saccharomyces, Torulospora, and Kluyveromyces) were selected to evaluate their fermentative performance and survival in a mixed culture on a medium based on Agave tequilana must, and their population kinetics was characterized using specific fluorescent in situ hybridization (FISH) probes in a qualitative and semi-quantitative analysis during fermentation. We observed that the mixture ratios of 0.1:1:1 or 1:1:1 (Saccharomyces:Kluyveromyces:Torulospora), maintained good fermentation productivities, with alcohol yields above 0.45 g/g, and allowed a high survival rate of the non-Saccharomyces strains during the fermentation process. Finally, mixed inoculum fermentations on A. tequilana must medium, including different Saccharomyces strains and the finally selected Torulospora and Kluyveromyces strains, showed the best production parameters in terms of ethanol, carbon dioxide, glycerol, and acetic acid values, as well as improved volatile metabolite profiles as compared to the pure cultures. All these data were used to propose a methodology of selection of strains to be used as a pure or mixed starter for tequila and mezcal fermentations, with high primary metabolite productivity and desired aromatic profile.

Removal of Phenols in Table Olive Processing Wastewater by Using a Mixed Inoculum of Candida boidinii and Bacillus pumilus: Effects of Inoculation Dynamics, Temperature, pH, and Effluent Age on the Abatement Efficiency

The main goal of this paper was to assess the ability of a combination of Candida boidinii and Bacillus pumilus to remove phenol in table olive processing water, as a function of some variables, like temperature, pH, a dilution of waste and the order of inoculation of the two microorganisms. At this purpose C. boidinii and B. pumilus were sequentially inoculated in two types of table olive processing water (fresh wastewater, FTOPW and wastewater stored for 3 months-aged wastewater, ATOPW). pH (6 and 9), temperature (10 and 35 °C) and dilution ratio (0, 1:1) were combined through a 2k fractional design. Data were modeled using two different approaches: Multifactorial Analysis of Variance (MANOVA) and multiple regression. A higher removal yield was achieved by inoculating B. pumilus prior to the yeast (192 vs. 127 mg/L); moreover, an increased efficiency was gained at 35 °C (mean removal of 200 mg/L). The use of two statistic approach suggested a different weight of variables; temperature was a global variable, that is a factor able to affect the yield of the process in all conditions. On the other hand, an alkaline pH could increase the removal of phenol at 10 °C (25–43%).

Selection Process of a Mixed Inoculum of Non-Saccharomyces Yeasts Isolated in the D.O.Ca. Rioja

The use of non-Saccharomyces yeasts in sequential fermentations with S. cerevisiae has been proposed to improve the organoleptic characteristics involved in the quality of wine. The present study set out to select a non-Saccharomyces inoculum from the D.O.Ca. Rioja for use in winemaking. Strains included in the study belonged to Torulaspora delbrueckii, Lachancea thermotolerans, Metschnikowia pulcherrima, Zygosaccharomyces bailii, Williopsis pratensis, Debaryomyces hansenii, Pichia kluyveri, Sporidiobolus salmonicolor, Candida spp., Cryptococcus spp. and two mixed inocula of Lachancea thermotolerans-Torulaspora delbrueckii in a 30/70 ratio. In the first stage of the process, SO2 resistance and presence of enzymatic activities related to wine aroma and wine color and fining (esterase, esterase-lipase, lipase, leucine arylamidase, valine arylamidase, cystine arylamidase, β-glucosidase, pectinase, cellulose, xylanase and glucanase) were studied. In the later stages, selection criteria such as fermentative behavior, aroma compound production or influence on phenolic compounds were studied in laboratory scale vinifications. Taking into account the results obtained in the different stages of the process, a mixed inoculum of Lachancea thermotolerans-Torulaspora delbrueckii in a 30/70 ratio was finally selected. This inoculum stood out for its high implantation capacity, the production of compounds of interest such as glycerol and lactic acid and the consequent modulation of wine acidity. Given these characteristics, the selected inoculum is suitable for the production of quality wines.

Optimizing Biocontrol Activity of Paenibacillus xylanexedens for Management of Hairy Root Disease in Tomato Grown in Hydroponic Greenhouses

Hairy root disease (HRD) caused by rhizogenic Agrobacterium biovar 1 strains affect tomato, cucumber, eggplant, and bell pepper grown in hydroponic greenhouses and can cause considerable yield losses worldwide. Recently, Paenibacillus xylanexedens strains (ST15.15/027 and AD117) with antagonistic activity against rhizogenic agrobacteria were identified. In this study, we present results of greenhouse trials of two consecutive growing seasons (2019 and 2020) to examine the potential of these two biocontrol organisms (BCOs) under practical conditions. BCO-treatment at a 107 colony forming units (CFU)/mL density resulted in a considerable reduction of the HRD infestation rate, confirming the biocontrol potential of the two P. xylanexedens strains. Results revealed that a single BCO strain (ST15.15/027) performed equally well as the mixed inoculum of both strains. The same level of biocontrol activity was even achieved when the BCO inoculum density was reduced to 105 CFU/mL. qPCR analysis further showed that Paenibacillus was still present in rockwool substrate near the end of both trials, indicating that they persist well in a rockwool environment and that application at the start of the trial is sufficient to protect tomato plants until the end of the trial. Altogether, these results are highly valuable for further optimization and exploitation of P. xylanexedens as a biocontrol product for the control of HRD in hydroponic greenhouses.

The Modulating Effect of Dietary Beta-Glucan Supplementation on Expression of Immune Response Genes of Broilers during a Coccidiosis Challenge

This study investigated the effects of a yeast-derived β-glucan (Auxoferm YGT) supplementation on mRNA expression of immune response genes in the spleen, thymus, and bursa of broiler chickens during a mixed Eimeria infection. Day (d)-old chicks (n = 1440) were fed diets containing 0% or 0.1% YGT. On d 8 post-hatch, half the replicate pens (n = 8) were challenged with a mixed inoculum of E. acervulina, E. maxima, and E. tenella. On d 10 and d 14 post-hatch, the spleen, thymus, and bursa were collected to evaluate mRNA abundance by quantitative real-time PCR. Data were analyzed using PROC GLIMMIX model (2-way interaction) and differences were established by LS-MEANS with significance reported at p ≤ 0.05. In spleen tissues at d 10, expression of interleukin (IL)-10 and inducible nitric oxide synthase (iNOS) were elevated in both 0.1% YGT-fed challenged and non-challenged birds. In thymus tissues at d 14, expression of IL-10, IL-17F, interferon (IFN)-γ, iNOS, and macrophage migration inhibitory factor (MIF) were elevated in challenged birds fed 0.1% YGT. In bursal tissues at d 10 and d 14, expression of IL-10, IFN-γ, iNOS (d 10 only), and MIF were elevated in 0.1% YGT-fed challenged and non-challenged birds. Dietary β-glucan supplementation to chicken diets modulated their immune response to the Eimeria challenge.

Mezcal as a Novel Source of Mixed Yeasts Inocula for Wine Fermentation

Mezcal yeasts were evaluated for their potential as grape-juice fermenters, characterizing their fermentation performance, both in terms of primary and volatile metabolites. Experiments were first carried-out in a semi-synthetic medium and then on grape juice, and population dynamics of the chosen mixed inoculum was assessed in grape juice. Accordingly, we initially tested 24 mezcal yeasts belonging to ten different species, and chose those that were more productive and stress tolerant for the mixed (dual) inoculum, having a final selection of three Saccharomyces cerevisiae strains (plus Fermichamp, a commercial wine strain) and three non-Saccharomyces strains, belonging to Kluyveromyces marxianus, Torulaspora delbrueckii, and Zygosaccharomyces bailii species. For the combination S. cerevisiae/T. delbrueckii (Sc/Td) mixed inoculum, we observed increasing isoamyl alcohol and phenyl ethyl acetate concentrations, as compared with the use of individual Saccharomyces strains, which resulted in a fruitier aroma profile. Alcohol final concentration was in average lower for the Sc/Td inoculum (fermentation power, FP, 13.6) as compared with the individual mezcal Saccharomyces strains (FP 14.3), and it was the highest when Td was co-cultured with the commercial strain (FP 14.6). Overall, our results show the feasibility of using yeasts isolated from mezcal as a novel source of inoculum for wine-type fermentation.