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Aerobic granular sludge (AGS) biotechnology has recently obtained considerable interest as a viable alternative to the activated sludge process (ASP) technique. This is because AGS can improve the performance of wastewater treatment and its significant capability for attaining sustainable growth. One of the main challenging issues for the aerobic granular sludge is the long startup time in wastewater treatment. This review presents a comprehensive analysis of AGS biotechnology, aiming specifically at its global adoption, startup duration, and granule stability. Aerobic granular sludge (AGS) is becoming more widespread globally. To ensure the successful implementation of AGS biotechnology, it is crucial to thoroughly identify the development of dense and stable granules, which is vital for the proper operation of wastewater treatment plants (WWTPs). Additionally, it offers a comprehensive summary of the latest advancements in the inoculum and polymer additives and their respective contributions to accelerate various processes through distinct mechanisms. In addition, this paper reviews the prevailing research patterns in the prompt initiation of rapid startup of AGS technology and outlines specific issues for future investigations.

The comprehensive management of organic urban solid waste is a concern due to its direct and indirect impact on the environment. Anaerobic Digestion (AD) has been recognized as an alternative and environmentally friendly technology for waste disposal, converting them into organic fertilizers and renewable energy. This research presents an experiment involving four reactors fed with household organic waste, three inoculated with canine, goat, and rabbit manure, and one without inoculum. The experiment was observed for 30 consecutive days to analyze the pH and temperature parameters involved in the AD process in domestic reactors. Statistical methodology, including one-way analysis of variance for assessing the effect of the type of inoculum, Tukey's simultaneous confidence intervals for mean differences, and 90 % confidence intervals for μ in temperature and manure, was utilized. Additionally, main effects analysis of the factors of average temperature and pH were conducted. The results of the one-factor experiment show that the type of inoculum does not significantly influence the variation in pH, while temperature remains relatively stable throughout the AD process. However, the analysis of main effects indicates that goat manure tends to stabilize the temperature with minimal variation, whereas variation is more heterogeneous in the other experiments.

This study aims to enhance biodegradation in WSP through the integrated application of thermal coating and inoculation. The integrated-coated concrete ponds (ICCPs) were designed as single-cell units of different aspect ratios and operated as a discontinuous-flow system. Settled sewage was first uniformly dosed in a constant head tank with mixed mesotrophic bacteria culture at 1.7 %, 3.3 %, and 5.0 % volume/volume ratio before being treated in the ponds. The result showed that it takes ten days to reduce BOD to 20 mg/L and 6 mg/L in the control pond and ICCP respectively. The heat energy causes the water temperature to increase due to the thermal capacity of the coated concrete surface maintaining a temperature condition in the pond system. Hence, this study was able to bridge the gap in the daily fluctuation of temperatures which influences microbial activity. Therefore, the effluent of the novel design was found suitable for irrigation purposes.

The inoculum has a crucial impact on bioreactor initialization and performance. However, there is currently a lack of guidance on selecting appropriate inocula for applications in environmental biotechnology. In this study, we applied microbial electrolysis cells (MECs) as models to investigate the differences in the functional potential of electroactive microorganisms (EAMs) within anodic biofilms developed from four different inocula (natural or artificial), using shotgun metagenomic techniques. We specifically focused on extracellular electron transfer (EET) function and stress resistance, which affect the performance and stability of MECs. Community profiling revealed that the family Geobacteraceae was the key EAM taxon in all biofilms, with Geobacter as the dominant genus. The c-type cytochrome gene imcH showed universal importance for Geobacteraceae EET and was utilized as a marker gene to evaluate the EET potential of EAMs. Additionally, stress response functional genes were used to assess the stress resistance potential of Geobacter species. Comparative analysis of imcH gene abundance revealed that EAMs with comparable overall EET potential could be enriched from artificial and natural inocula (P > 0.05). However, quantification of stress response gene copy numbers in the genomes demonstrated that EAMs originating from natural inocula possessed superior stress resistance potential (196 vs. 163). Overall, this study provides novel perspectives on the inoculum effect in bioreactors and offers theoretical guidance for selecting inoculum in environmental engineering applications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Availability of fossil fuels is increasingly limited, so the search for alternative fuels is important. In accordance with the current scenario, bioenergy research emphasizes the bioethanol production from plants that are abundant and available throughout the year, such as palm sap. The palm sap is a type of palm tree that grows in tropical forests, particularly in South Asia and Southeast Asia. More than 3000 species of palm exist, and they are categorised as multipurpose trees because they can be used as raw materials for various products, such as sugar, fermented drinks, syrup, palm wine, vinegar, alcohol and bioethanol. This study was aimed to examine bioethanol production from palm sap through optimization of Saccharomyces cerevisiae as an inoculum. The research sample was obtained from local farmers in Pusuk, Lombok Island, West Nusa Tenggara, and Indonesia. The research parameters included the pH change, colour, and ethanol content. The results showed that the pH change of the palm sap during storage was caused by the growth of microorganisms to produce organic acids by releasing hydrogen ions. As pH decreased, L∗ and b∗ values also decreased significantly, but not significantly change in a∗ values was observed. Glucose changed to ethanol during fermentation. The higher the percentage of inoculum used, the higher the volume of ethanol obtained. The bioethanol content of palm sap prior to fermentation was 32.3%, and it increased to 75.6% after 24 hours of incubation.

This paper proposes and develops a protocol for measuring the aerobic granulation potential of sludge, aiming to provide an affordable and simple alternative that can facilitate the development of aerobic granulation technology. In this sense, the protocol comprises a set of parameters and considerations that interact to create a controlled environment and stimulate cell population clustering. All of this is done in the context of procedural simplicity, low cost, and the speed at which results are obtained. The protocol is essentially a three-stage method: preparation of the substrate, adaptation of the inoculum, and implementation of the protocol. Simple parameters were measured to evaluate the granulation process: SVI, settling velocity, and morphological parameters. The protocol was validated according to optimal ranges and criteria previously established in the literature. For this purpose, an activated sludge inoculum from a domestic wastewater treatment plant was submitted to the protocol, obtaining an optimal response of the biomass (SVI5 =13.90 mL g−1, settling velocity= 25,79 m h−1, Diameter > 0.2 mm) in a relatively short time (7 d). The results show that this protocol can constitute a tool for evaluation and decision-making using traditional laboratory equipment and is applicable at different scales.

A relationship between the infecting dose and the risk of disease severity has not been demonstrated for SARS-CoV-2 infection. Here, we report three clusters of individuals that were potentially exposed to distinct inoculum in Madrid. Overall each group developed divergent clinical forms of COVID-19. Our data support that a greater viral inoculum at the time of SARS-CoV-2 exposure might determine a higher risk of severe COVID-19.

Biodegradation is the breakdown of complex organic compounds into simpler molecules like carbon dioxide and water by microorganisms like bacteria and fungi. Biodegradation studies of pharmaceuticals are initially done to assess which pharmaceuticals are persistent in the environment. Whole pharmaceuticals or their metabolites are excreted from the human body via urine or fecal matter after administration. These go into the Wastewater Treatment Plants (WWTP) and are later released into the environment with the treated wastewater. Recent studies have reported a number of pharmaceuticals in the ecosystem and the effects of these on non-target species has become an issue of environmental concern. The biodegradation studies of eight pharmaceuticals were carried out in this research. The choice of pharmaceuticals was based on the most commonly prescribed medications at the University of Zambia (UNZA) Clinic in seven therapeutic groups: anti-hypertensives, antibiotic, antimalarial drugs, anti-tuberculosis, antihelminthics, antifungals and antiretroviral drugs. The biodegradability tests were carried out using a modified carbon dioxide evolution method (modified Sturm test). The inoculum was derived from the secondary effluent of the UNZA WWTP plant and Dextrose monohydrate was used as a system control. Using this guideline, the system control, dextrose monohydrate biodegraded 77 ± 0.270% in seven days. All the pharmaceuticals except ciprofloxacin were found to be non-biodegradable: Atenolol degraded 6.8 ± 0.026%, ketoconazole degraded 1.0 ± 0.003%, isoniazid/rifampicin degraded 0.8 ± 0.003%, mebendazole degraded 13.0 ± 0.050%, nevirapine degraded 1.3 ± 0.005%, pen-v degraded 1.0 ± 0.004% and quinine sulfate degraded 1.8 ± 0.008%. Ciprofloxacin showed a negative carbon dioxide evolution and it was noted that bacteria were not viable as the drug proved to be very potent against bacterial strains in the inoculum used.

The natural microbes are potential contributor to build up soil nitrogen through transformation of molecular nitrogen to plant available forms. Therefore, in the present study, we investigated the contribution of biofertilizer to reduce the synthetic nitrogen application without deteriorating the yield and forage quality. The supplementary nitrogen rates included 0, 40, 80 and 120 kg ha−1 and the seed inoculation was carried out with the mixture of Azospirillum + Azotobacter spp. The experiment was laid out in randomized complete block design with factorial arrangement. The results indicated that organic matter contents and ether extractable fat were negatively associated with both nitrogen and inoculation factors. The inoculation produced 6.58%, 9.58%, 2.51%, 16.94%, 10.26%, 17.59%, 14.02%, 33.81% and 66.18% more No. tillers, plant height, leaf to stem ratio, dry matter yield, mineral matter contents, crude fibre, crude protein, crude protein yield and total digestible crude protein yield, respectively over uninoculation. The interactive effects indicated that inoculation alone without nitrogen application produced 19.16% and 6.87% more dry matter yield and crude protein (%), respectively. The beneficiary effects of biofertilizers on growth and dry matter of oat were more pronounced at intermediate level of inorganic nitrogen which was gradually decreased at higher nitrogen levels. The CP, CPY and DCPY achieved with inoculation alone were statistically equivalent to plots fertilized with 0 and 40 kg N ha−1. It is clear that plots sown with inoculated seeds must be fertilized with 80 kg N to produce higher dry matter and economic returns. However, the highest protein contents in dry matter were recorded with highest fertilization level along with inoculation. By giving due attention to stimulatory effects of bacterial species in the present study, it is therefore, recommended to integrate the use of biofertilizers with supplemental dose of synthetic nitrogen source to sustain crop production.

Succinic acid (SA) has been produced from rice straw (RS) and sugarcane bagasse (SB) as low-cost feedstocks in this study through sequential peracetic acid (PA) and alkaline peroxide (AP) pretreatment assisted by ultrasound and pre-hydrolysis followed by simultaneous saccharification and fermentation (PSSF). The effect of yeast extract (YE) concentration, inoculum concentration, and biomass type on SA production was investigated. The results showed that SA production from RS and SB was significantly affected by the YE concentration. Final concentration and yield of SA produced were significantly increased along with the increasing of YE concentration. Moreover, inoculum concentration significantly affected the SA production from SB. Higher inoculum concentration led to higher SA production. On the other hand, SA production from RS was not significantly affected by the inoculum concentration. Using RS as the feedstock, the highest SA production was achieved on the medium containing 15 g/L YE with 5 % v/v inoculum, obtaining SA concentration and yield of 3.64 ± 0.1 g/L and 0.18 ± 0.05 g/g biomass, respectively. Meanwhile, the highest SA production from SB was acquired on the medium containing 10 g/L YE with 7.5 % v/v inoculum, resulting SA concentration and yield of 5.1 ± 0.1 g/L and 0.25 ± 0.05 g/g biomass, respectively. This study suggested that RS and SB are potential to be used as low-cost feedstocks for sustainable and environmentally friendly SA production through ultrasonic-assisted PA and AP pretreatment and PSSF., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Arbuscular mycorrhizal (AM) fungi play a key role in determining ecosystem functionality. Understanding how diversity in the fungal community affects plant productivity is therefore an important question in ecology. Current research has focused on understanding the role of functional complementarity in the fungal community when the host plant faces multiple stress factors. Fewer studies, however, have investigated how variation in traits affecting nutrient exchange can impact the plant growth dynamics, even in the absence of environmental stressors. Combining experimental data and a mathematical model based on ordinary differential equations, we investigate the role played by carbon sink strength on plant productivity. We simulate and measure plant growth over time when the plant is associated with two fungal isolates with different carbon sink strength, and when the plant is in pairwise association with each of the isolates alone. Overall, our theoretical as well as our experimental results show that co-inoculation with fungi with different carbon sink strength can induce positive non-additive effects (or synergistic effects) in plant productivity. Fungi with high carbon sink strength are able to quickly establish a fungal community and increase the nutrient supply to the plant, with a consequent positive impact on plant growth rate. On the other side, fungi with low carbon sink strength inflict lower carbon costs to the host plant, and support maximal plant productivity once plant biomass is large. As AM fungi are widely used as organic fertilizers worldwide, our findings have important implications for restoration ecology and agricultural management., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

The aim of this work was to provide solid proofs regarding the achievement of "steady-state conditions", which means that the performance of the anaerobic digester is representative of the applied environmental conditions. For this reason, we investigated how, starting from different inoculum sources (i.e., municipal wastewater treatment, bio-waste treatment, and agricultural waste biogas plant), the microbial community adapted to the operational parameters and led to stable biogas production in thermophilic digesters treating the same influent feedstock. The results revealed that the different system achieved similar process performance and microbial community structure after a period that was equal to four hydraulic retention times, approved by a constant pH of 7.89 ± 0.08, 7.92 ± 0.05 and 7.85 ± 0.08, respectively, and stable TAN concentration of 1500 mg/L. Moreover, it was found that the microbial composition of the inocula was a key factor for the speed of achieving stable process performance; thus, a pre-adapted to the influent feedstock inoculum can shorten the stabilization process. On the contrary, after long term reactor operation, the microbial structure was shaped according to the chemical composition of the influent feedstock. The results of the study can also be used as a guide in future researches on anaerobic degradation, particularly in determining the time interval of an experiment to reflect changes in the microbial community of anaerobic digester., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Abstract: Salmonella enterica has been increasingly linked to outbreaks involving consumption of fresh produce. Although researchers have identified genes whose products are involved in mediating S. enterica-plant interactions, the use of various experimental approaches, serovars, and plant types has generated variable and conflicting data. The purpose of this study was to determine whether conditions under which inocula are prepared for in vitro plant interaction studies influence the outcome of these studies. Seven S. enterica serovars were grown in media that differed in salinity and physical state with incubation at 25 or 37°C. These cultures were then used to inoculate red leaf lettuce, and adherent microbes were subsequently recovered. Although all Salmonella serovars were influenced by inoculum preparation conditions, the amount of variation differed. Analysis of pooled serovar data revealed that inocula prepared from either agar plates or biphasic cultures had higher levels of interaction with red leaf lettuce than those prepared from broth cultures. Incubation at 37°C enhanced adherence after 30 s or 5 days of contact time, and adherence after 1 h of contact time was increased in low-salt medium. Broth inoculum cultures were highly influenced by medium salinity and incubation temperature, whereas plate and biphasic inoculum cultures were only minimally affected. Therefore, inocula prepared from bacteria grown on plates or in biphasic culture would be most suitable for evaluation of strategies used to interfere with plant-Salmonella interactions. However, pooled data mask serovar-specific responses, and care should be taken when extrapolating these findings to individual serovars. The previous association of a serovar with outbreaks involving leafy greens was not correlated with levels of interaction with red leaf lettuce, suggesting that the occurrence of these serovars in or on these commodities does not reflect their fitness in the plant environment., (Copyright ©, International Association for Food Protection.)

Methane gas is widespread in natural environments and anaerobic wastewater treatment sites, bringing the risk of the greenhouse effect and energy loss if left unmanaged. A methane-fueled microbial fuel cell (MFC) can convert methane to electricity under mild condition, but faced difficulties in startup. In this study, the new startup strategy and operation performance for methane-fueled MFCs were investigated. After the pre-cultivation of formate-acclimating electroactive culture, the methane-fueled MFC was successfully started up in a short time of 53 d. Increasing concentrations of molybdenum and tungsten in medium facilitated both methane consumption and electricity generation. Under the optimal condition (batch duration of 11 h, 30 °C, pH 7 buffered by phosphate buffer solution), the methane-fueled MFC achieved the maximum power density of 166 mW/m 3 , a coulomb production of 6.58 ± 0.07C/batch, a CE of 27.4 ± 0.4% and a methane consumption of 31.2 ± 0.3 μmol/batch. This work explored a suitable inoculum (formate-acclimating electroactive culture) for methane-fueled MFCs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

A relationship between the infecting dose and the risk of disease severity has not been demonstrated for SARS-CoV-2 infection. Here, we report three clusters of individuals that were potentially exposed to distinct inoculum in Madrid. Overall each group developed divergent clinical forms of COVID-19. Our data support that a greater viral inoculum at the time of SARS-CoV-2 exposure might determine a higher risk of severe COVID-19., (Copyright © 2020 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Rapid assessment of fungal growth and screening antifungal compounds, such as nanoparticles (NPs), for effectiveness is a challenging procedure because no primary standards exist as they do for yeasts and bacteria. Because fungi do not grow as single cells, but as hyphal filaments, they cannot be quantified by the usual enumeration techniques used in bacteriology. The growth of three postharvest fungal isolates ( Alternaria alternata, Rhizopus stolonifer, and Botrytis cinerea) was investigated at different inoculum concentrations and in three nutrient media (Sabouraud dextrose agar, potato dextrose agar, and yeast extract dextrose agar [YED]) with a turbidimetric assay. Sequential measurements were performed to generate optical density versus time plots, whereas the growth responses were expressed quantitatively as the generated trapezoidal area. YED medium showed the lowest variation among replicated experiments; potato dextrose agar showed the next lowest, but there was no significant difference. The inoculum size had a minimal effect on the variation of the fungal dynamics. Microscopic assessment of the fungal growth confirmed that YED medium allowed the most homogeneous development of the studied fungi. Therefore, we developed a rapid and reliable technique to evaluate the efficacy of novel antifungal compounds such as zinc oxide NPs. Turbidimetric assessment showed that these NPs were able to inhibit the growth of all three isolates. A. alternata and B. cinerea did not show a significant difference in the level of inhibition at 15 mM, whereas R. stolonifer showed the highest inhibition at the same concentration.

The factors that affect microbial community assembly and its effects on the performance of bioelectrochemical systems are poorly understood. Sixteen microbial fuel cell (MFC) reactors were set up to test the importance of inoculum, temperature and substrate: Arctic soil versus wastewater as inoculum; warm (26.5°C) versus cold (7.5°C) temperature; and acetate versus wastewater as substrate. Substrate was the dominant factor in determining performance and diversity: unexpectedly the simple electrogenic substrate delivered a higher diversity than a complex wastewater. Furthermore, in acetate fed reactors, diversity did not correlate with performance, yet in wastewater fed ones it did, with greater diversity sustaining higher power densities and coulombic efficiencies. Temperature had only a minor effect on power density, (Q 10 : 2 and 1.2 for acetate and wastewater respectively): this is surprising given the well-known temperature sensitivity of anaerobic bioreactors. Reactors were able to operate at low temperature with real wastewater without the need for specialised inocula; it is speculated that MFC biofilms may have a self-heating effect. Importantly, the warm acetate fed reactors in this study did not act as direct model for cold wastewater fed systems. Application of this technology will encompass use of real wastewater at ambient temperatures., (Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.)

Provision of digestate with satisfactory biosafety is critical to land application of digestate and to the anaerobic digestion approach to treating manure and food waste (FW). No studies have been conducted on digestate biosafety in dry co-digestion systems. The aim of this study was to assess the inactivation efficiency and possible inactivation mechanism for three enteric indicator bacteria and the system stability during dry mesophilic anaerobic co-digestion of FW and pig manure (PM). The effects of two different inocula were examined at a rate of 50% based on volatile solids (VS): digestate taken from existing dry co-digestion digesters and dewatered anaerobic sludge from a local wastewater treatment plant. The FW/PM ratios of 50:50 and 75:25 on a VS basis were also assessed. The results showed that using digestate as the inoculum and a FW/PM ratio of 50:50 led to stable dry co-digestion, with the specific methane yield (SMY) of 252mL/gVS added . Total volatile fatty acid (VFA) concentration was a significant inhibition factor for methane production during dry co-digestion (P<0.001). The data also showed that dry co-digestion of FW and PM effectively inactivated enteric indicator bacteria. E. coli and total coliforms counts decreased below the limit of detection (LOD, 10 2 CFU/g) within 4-7days, with free VFA identified as a significant inactivation factor. Enterococci were more resistant but nonetheless the counts decreased below the LOD within 12days in the digestate inoculum systems and 26-31days in the sludge inoculum systems. The residence time was the most significant inactivation factor for enterococci, with the free VFA concentration playing a secondary role at high FW/PM ratio in the sludge inoculum system. In conclusion, digestate as inoculum and the FW/PM ratio of 50:50 were preferable operation conditions to realize system stability, methane production and enteric indicator bacteria inactivation., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Alicyclobacillus acidoterrestris is a thermotolerant bacterium able to grow in fruit juices and drinks, as the spoilage by Alicyclobacillus in the final product does not product any gas, but leads to a "medicine flavor" due to the formation of guaicol. Also, its detection is a challenge for the quality control departments, because it takes several days to get the results of traditional microbiology methods. This study aimed at developing a more accurate electrical impedance technique for the detection of A. acidoterrestris in concentrated apple juice. Samples of apple juice were inoculated with A. acidoterrestris spores isolated from a peach and grape juice. For the spore germination, several heat-shock treatments were tested (80 °C/10 min, 70 °C/20 min and 60 °C/30 min). Direct and indirect electrical impedance was applied to detect and quantify the microorganism in the inoculated apple juice, using BAT broth and Bimedia 002A (pH 4). The 80 °C/10 min treatment was selected for spore activation. The valid electrical impedance technique was the indirect method in BAT broth, which measured the changes in the impedance through the formation of CO 2 . In addition, a positive correlation (r = 0.98, R 2  = 0.97) was observed between the classical microbiology (BAM agar) and the indirect impedance method., (Copyright © 2017 Elsevier Ltd. All rights reserved.)