•Three inocula have been tested systematically with four different substrates.•If inoculum’s origin impacts the performance in BMP test depend on the substrate.•Methane yield was not significantly ...influenced by the inoculum except for cellulose.•Degradation velocity was highest for the inoculum adapted to the substrate.
The impact of the inoculum’s origin on the methane yield in Biochemical Methane Potential (BMP) tests was investigated. The three most commonly applied inocula were chosen, originating from (i) a digester of a wastewater treatment plant, (ii) an agricultural biogas plant treating manure and energy crops, and (iii) a biowaste treatment plant. The performance of each inoculum was tested with four different substrates, namely sewage sludge, dried whole crop maize, food waste, and microcrystalline cellulose as a typical reference material. The results revealed that the choice of inoculum had no significant impact on the specific methane yield of the tested substrates except for cellulose. Still, the specific methane production rate was significantly influenced by the choice of the inoculum especially for sewage sludge, but also for food waste and cellulose, whereas it became clear that an inoculum adapted to a substrate is beneficial for a speedy digestion.
This study evaluated the key role of inoculum in mesophilic anaerobic digestion (AD) of Organic Fraction of Municipal Solid Waste. The effect of two different inocula, the mesophilic digestate of ...wastewater activated sludge (WAS) and the mesophilic digestate of cow-agriculture sludge (CAS), at three different substrate: inoculum ratios (1:2, 1:1 and 2:1) at three different incubation times (0, 5 and 10 d) were studied in batch feeding reactor for a total of 18 AD configurations. The AD configurations were study through specific biogas and methane productions, first order disintegration kinetics, Gompertz modified study and energy sustainable index. A multi criteria decision aid outranked the 18 AD configurations tested. The study proved that the AD performed with inoculum CAS incubated for 10 d at 2:1 substrate inoculum ratio reached the highest biogas yield and methene content equal to 997.81 NL/kgVS and 70.00% v/v.
•Optimisation of anaerobic digestion (AD) to increase methane content up to 70 %v/v.•AD performed with inoculum incubated 5–10 d can work at higher substrate: inoculum mass ratio.•AD performed with inoculum incubated for 5 and 10 d reaches ESI higher than 1.•For inocula incubated for 5–10 d the kd is higher (0.49 d−1).•The lag phase is reduced from 4 to 0 d with incubated inoculum.
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•Acidogenesis of citrus waste was better at controlled pH 6 at low substrate loading.•Addition of inhibitor for methanogens did not significantly increase VFA yield.•The presence of ...initial O2 did not negatively impact the VFA yield.•D-Limonene and peel oil also can also inhibit acidification of citrus waste.
Citrus waste from e.g., juice production is a potential substrate for anaerobic digestion (AD). However, due to the toxic citrus peel oil content, citrus waste has several challenges in biogas production. Hence, volatile fatty acids (VFAs) are very interesting intermediate products of AD. This paper was aimed to investigate VFA production from citrus wastes by boosting its production and inhibiting methane formation. Therefore, the effects of inoculum to substrate ratio (ISR), O2 presence, pH, and inhibitor for methanogens, in VFA production from citrus waste through acidification process were studied. The addition of 2 g/L methanogens inhibitor and the presence of O2 in the reactors were able to reduce methane production. The highest yield of VFA (0.793 g VFA/g VSadded) was achieved at controlled pH at 6 and low substrate loading (ISR 1:1). Acetic acid (32%), caproic acid (21%), and butyric acid (15%) dominate the VFA composition in this condition.
Soil salinity is a limiting factor in crop productivity. Inoculating crops with microorganisms adapted to salt stress is an alternative to increasing plant salinity tolerance. Few studies have ...simultaneously propagated arbuscular mycorrhizal fungi (AMF) and dark septate fungi (DSF) using different sources of native inoculum from halophyte plants and evaluated their effectiveness. In alfalfa plants as trap culture, this study assessed the infectivity of 38 microbial consortia native from rhizosphere soil (19) or roots (19) from six halophyte plants, as well as their effectiveness in mitigating salinity stress. Inoculation with soil resulted in 26–56% colonization by AMF and 12–32% by DSF. Root inoculation produced 10–56% and 8–24% colonization by AMF and DSF, respectively. There was no difference in the number of spores of AMF produced with both inoculum types. The effective consortia were selected based on low Na but high P and K shoot concentrations that are variable and are relevant for plant nutrition and salt stress mitigation. This microbial consortia selection may be a novel and applicable model, which would allow the production of native microbial inoculants adapted to salinity to diminish the harmful effects of salinity stress in glycophyte plants in the context of sustainable agriculture.
Intraspecies variability in fungal growth and mycotoxin production has important implications for food safety. Using the Bioscreen C we have examined spectrophotometrically intraspecies variability ...of A. flavus using 10 isolates under different environments, including temperature shifts, in terms of growth and aflatoxin B1 (AFB1) production. Five high and five low AFB1 producers were examined. The study was conducted at 5 isothermal conditions (from 15 to 37 °C) and 4 dynamic scenarios (between 15 and 30 °C). The experiments were carried out in a semisolid YES medium at 0.92 aw and two inoculum levels, 102 and 103 spores/mL. The Time to Detection (TTD) of growth initiation was determined and modelled as a function of temperature through a polynomial equation and the model was used to predict TTD under temperature upshifts conditions using a novel approach. The results obtained in this study have shown that a model can be developed to describe the effect of temperature upshifts on the TTD for all the studied isolates and inoculum levels. Isolate variability increased as the growth conditions became more stressful and with a lower inoculum level. Inoculum level affected the intraspecies variability but not the repeatability of the experiments. In dynamic conditions, isolate responses depended both on the temperature shift and, predominantly, the final temperature level. AFB1 production was highly variable among the isolates and greatly depended on temperature (optimum temperature at 30–35 °C) and inoculum levels, with often higher production with lower inoculum. This suggests that, from an ecological point of view, the potential isolate variability and interaction with dynamic conditions should be taken into account in developing strategies to control growth and predicting mycotoxin risks by mycotoxigenic fungi.
•Time to growth detection (TTD) based on optical density was estimated for ten A. flavus isolates.•Intraspecies differences in TTD were small, but high for aflatoxin production.•Intraspecific differences were higher at lower temperature and inoculum levels.•At lower inoculum size a longer TTD but sometimes higher aflatoxin accumulation was obtained.•TTD at variable temperature was successfully predicted from models at constant temperature.
Inoculum effect of antimicrobial peptides Loffredo, Maria Rosa; Savini, Filippo; Bobone, Sara ...
Proceedings of the National Academy of Sciences - PNAS,
05/2021, Letnik:
118, Številka:
21
Journal Article
Recenzirano
Odprti dostop
Significance
Bacterial drug resistance is a crucial threat to global health, and antimicrobials with novel mechanisms of action are urgently needed. Antimicrobial peptides are natural molecules that ...kill bacteria mostly by perturbing their membranes, and they constitute promising compounds for fighting resistant microbes. Their activity is normally tested under standardized conditions of bacterial density. However, the bacterial load in clinically relevant infections varies by many orders of magnitude. Here, we show that the minimum peptide concentration needed for bacterial growth inhibition can vary by more than 100-fold with an increase in the density of cells in the initial inoculum of the assay (a phenomenon termed the “inoculum effect”). These findings question the utility of the currently used activity screening assays.
The activity of many antibiotics depends on the initial density of cells used in bacterial growth inhibition assays. This phenomenon, termed the inoculum effect, can have important consequences for the therapeutic efficacy of the drugs, because bacterial loads vary by several orders of magnitude in clinically relevant infections. Antimicrobial peptides are a promising class of molecules in the fight against drug-resistant bacteria because they act mainly by perturbing the cell membranes rather than by inhibiting intracellular targets. Here, we report a systematic characterization of the inoculum effect for this class of antibacterial compounds. Minimum inhibitory concentration values were measured for 13 peptides (including all-D enantiomers) and peptidomimetics, covering more than seven orders of magnitude in inoculated cell density. In most cases, the inoculum effect was significant for cell densities above the standard inoculum of 5 × 10
5
cells/mL, while for lower densities the active concentrations remained essentially constant, with values in the micromolar range. In the case of membrane-active peptides, these data can be rationalized by considering a simple model, taking into account peptide–cell association, and hypothesizing that a threshold number of cell-bound peptide molecules is required in order to cause bacterial killing. The observed effect questions the clinical utility of activity and selectivity determinations performed at a fixed, standardized cell density. A routine evaluation of the dependence of the activity of antimicrobial peptides and peptidomimetics on the inoculum should be considered.
The observed MIC may depend on the number of bacteria initially inoculated into the assay. This phenomenon is termed the inoculum effect (IE) and is often most pronounced for β-lactams in strains ...expressing β-lactamase enzymes. The Clinical and Laboratory Standards Institute (CLSI)-recommended inoculum is 5 × 10
CFU ml
with an acceptable range of 2 × 10
to 8 × 10
CFU ml
IE testing is typically performed using an inoculum 100-fold greater than the CLSI-recommended inoculum. Therefore, it remains unknown whether the IE influences MICs during testing performed according to CLSI guidelines. Here, we utilized inkjet printing technology to test the IE on cefepime, meropenem, and ceftazidime-avibactam. First, we determined that the inkjet dispense volume correlated well with the number of bacteria delivered to microwells in 2-fold (
= 0.99) or 1.1-fold (
= 0.98) serial dilutions. We then quantified the IE by dispensing orthogonal titrations of bacterial cells and antibiotics. For cefepime-resistant and susceptible dose-dependent strains, a 2-fold increase in inoculum resulted in a 1.6 log
-fold increase in MIC. For carbapenemase-producing strains, each 2-fold reduction in inoculum resulted in a 1.26 log
-fold reduction in meropenem MIC. At the lower end of the CLSI-allowable inoculum range, minor error rates of 34.8% were observed for meropenem when testing a resistant-strain set. Ceftazidime-avibactam was not subject to an appreciable IE. Our results suggest that IE is sufficiently pronounced for meropenem and cefepime in multidrug-resistant Gram-negative pathogens to affect categorical interpretations during standard laboratory testing.
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