Anaerobic Incubation Research Articles

Change in antimicrobial susceptibility of Listeria spp. in response to stress conditions

Listeria species are exposed to various stressors throughout the food chain, which are crucial for microbe mitigation strategy in the food industry. However, the survival capabilities and development of antimicrobial resistance by Listeria spp. under different food processing environments (FPEs) stressors are not yet well understood. Hence, this study aims to determine the difference in survivability and antimicrobial susceptibility of L. monocytogenes (Lm) and other Listeria species (non-Lm) strains exposed to different FPEs stressors, including heat, acidic and alkaline pH, UV irradiation, and osmotic stress. For this, a collection of 11 Lm and 10 non-Lm strains were used to conduct experiments. This study showed that Lm strains were relatively more tolerant to environmental stresses than non-Lm strains (p > 0.05). Additionally, the evaluation of stress-induced resistance toward antimicrobials showed that anaerobic incubation, after exposition to environmental stresses, rendered Lm and non-Lm more resistant to antimicrobial agents than aerobic incubation. Furthermore, the study observed that different stressors induced an increase in minimum inhibitory concentrations (MICs) of certain antimicrobials. Specifically, heat stress persuaded an increase in MICs of tetracycline under aerobic incubation, and gentamicin and ciprofloxacin under anaerobic incubation. Acidic/alkaline pH induced an increase in MICs of gentamicin, ciprofloxacin, and trimethoprim-sulfamethoxazole, especially under anaerobic incubation. However, UV stress induced increase in MICs of tetracycline and trimethoprim-sulfamethoxazole under aerobic incubation and gentamicin, ciprofloxacin, and trimethoprim-sulfamethoxazole under anaerobic incubation. Additionally, osmotic stress induced an increase in MICs of tetracycline and ampicillin under aerobic incubation and gentamicin, tetracycline, and trimethoprim-sulfamethoxazole under anaerobic incubation. Collectively, this study highlights that stress tolerance may contribute to the predominance of Listeria species among FPEs and induce the development of antimicrobial resistance even without antibiotic selection pressure. The findings of this study may guide updated strategies to mitigate Listeria species in the food industry.

Cutibacterium acnes in spine tissue: characteristics and outcomes of non-hardware-associated vertebral osteomyelitis.

Cutibacterium acnes isolation from spine tissue can be challenging because the organism can represent a contaminant. There is a paucity of data regarding the role of C.acnes in non-hardware-associated vertebral osteomyelitis (VO). Herein we evaluate the clinical and microbiological characteristics, treatment, and outcome of patients with C.acnes VO. Data were retrospectively collected from adults with a positive spine culture for C.acnes at Mayo Clinic, Rochester (MN), from 2011 to 2021. Patients with spinal hardware and polymicrobial infections were excluded. Of the subjects, 16 showed radiological and clinical findings of VO: 87.5 % were male, the average age was 58years ( SD), and back pain was the predominant symptom. Of the lesions, 89.5 % involved the thoracic spine. Of the subjects, 69 % had experienced an antecedent event at the site of VO. In five subjects, C.acnes was isolated after 7 d of anaerobic culture incubation. Thirteen subjects were treated with parenteral -lactams, and three with oral antimicrobials, without any evidence of recurrence. Twenty-one subjects were not treated for VO, as C.acnes was considered a contaminant; at follow-up, none had evidence of progressive disease. C.acnes should be part of microbiological differential diagnosis in patients with suspected VO, especially in the context of a prior spinal procedure. Anaerobic spine cultures should undergo prolonged incubation to enable recovery of C.acnes. C.acnes VO may be managed with oral or parenteral antimicrobial therapy. Without clinical and radiological evidence of VO, a single positive culture of C.acnes from spine tissue frequently represents contaminants.

Anaerobic oxidation of methane does not attenuate methane emissions from thermokarst lakes

AbstractThe ongoing global temperature rise enhances permafrost thaw in the Arctic, allowing Pleistocene‐aged frozen soil organic matter to become available for microbial degradation and production of greenhouse gases, particularly methane. Here, we examined the extent and mechanism of anaerobic oxidation of methane (AOM) in the sediments of four interior Alaska thermokarst lakes, which formed and continue to expand as a result of ice‐rich permafrost thaw. In cores of surface (~ 1 m) lake sediments we quantified methane production (methanogenesis) and AOM rates using anaerobic incubation experiments in low (4°C) and high (16°C) temperatures. Methanogenesis rates were measured by the accumulation of methane over ~ 90 d, whereas AOM rates were measured by adding labeled‐13CH4 and measuring the produced dissolved inorganic 13C. Our results demonstrate that while methanogenesis was vigorous in these anoxic sediments, AOM was lower by two orders of magnitude. In almost all sediment depths and temperatures, AOM rates remained less than 2% of the methanogenesis rates. Experimental evidence indicates that the AOM is strongly related to methanogens, as the addition of a methanogens' inhibitor prevented AOM. Variety of electron acceptor additions did not stimulate AOM, and methanotrophs were scarcely detected. These observations suggest that the AOM signals in the incubation experiments might be a result of enzymatic reversibility (“back‐flux”) during CH4 production, rather than thermodynamically favorable AOM. Regardless of the mechanism, the quantitative results show that near surface (< 1‐m) thermokarst sediments in interior Alaska have little to no buffer mechanisms capable of attenuating methane production in a warming climate.

A joint role of iron oxide and temperature for methane production and methanogenic community in paddy soils

Microbial methanogenesis in paddy soils contributes approximately one-fifth of global anthropogenic methane (CH4) release, creating severe negative climate feedback. The dynamics of iron oxides in flooded paddy fields, as well as temperature changes, have an impact on CH4 production. However, the relationship between the two, as well as their interactive mechanisms in influencing CH4 production and microbial communities in paddy soils, is not yet clear. Therefore, we investigated the interactive effects of temperature (15, 25 and 35 °C) and iron oxide (ferrihydrite) on CH4 production and methanogenic community structure in two Chinese paddy soils, one subtropical and one temperate. We characterized shifts in microbial communities using high-throughput sequencing of bacteria, archaea and methanogenic mcrA genes following an anaerobic incubation for 90 days. Our results showed that temperature significantly promoted Fe reduction in paddy soils, and both together regulated CH4 production by reducing the number of methanogens. Additionally, the temperature sensitivity of methanogenesis was higher in the temperate than in the subtropical paddy soil. We also demonstrated that ferrihydrite can inhibit methanogenesis by reducing the relative abundance of Methanosarcinales and altering the community patterns of methanogens in these paddy soils. Likewise, increases in temperature changes the relative abundance of Methanosarcinales and Methanobacteriales, promoting methanogenesis. Overall, our results provide new insights into the role of iron oxides and temperature in regulating greenhouse gas emissions from paddy soils.

Microbiota-derived short-chain fatty acids mediate the effects of dengzhan shengmai in ameliorating cerebral ischemia via the gut-brain axis.

Dengzhan shengmai (DZSM) formula, composed of four herbal medicines (Erigeron breviscapus, Panax ginseng, Schisandra chinensis, and Ophiopogon japonicus), is widely used in the recovery period of ischemic cerebrovascular diseases; however, the associated molecular mechanism remains unclear. The purpose of this study was to uncover the links between the microbiota-gut-brain axis and the efficacy of DZSM in ameliorating cerebral ischemic diseases. The effects of DZSM on the gut microbiota community and bacteria-derived short-chain fatty acid (SCFA) production were evaluated in vivo using a rat model of cerebral ischemia and in vitro through the anaerobic incubation with fresh feces derived from model animals. Subsequently, the mechanism underlying the role of SCFAs in the DZSM-mediated treatment of cerebral ischemia was explored. We found that DZSM treatment significantly altered the composition of the gut microbiota and markedly enhanced SCFA production. The consequent increase in SCFA levels led to the upregulation of the expression of monocarboxylate transporters and facilitated the transportation of intestinal SCFAs into the brain, thereby inhibiting the apoptosis of neurocytes via the regulation of the PI3K/AKT/caspase-3 pathway. The increased intestinal SCFA levels also contributed to the repair of the 2VO-induced disruption of gut barrier integrity and inhibited the translocation of lipopolysaccharide from the intestine to the brain, thus attenuating neuroinflammation. Consequently, cerebral neuropathy and oxidative stress were significantly improved in 2VO model rats, leading to the amelioration of cerebral ischemia-induced cognitive dysfunction. Finally, fecal microbiota transplantation could reproduce the beneficial effects of DZSM on SCFA production and cerebral ischemia. Our findings suggested that SCFAs mediate the effects of DZSM in ameliorating cerebral ischemia via the gut microbiota-gut-brain axis.

Co-incorporation of Chinese milk vetch (Astragalus sinicus L.) and chemical fertilizers alters microbial functional genes supporting short-time scale positive nitrogen priming effects in paddy soils

Nitrogen (N) priming is a microbially mediated biochemical process as affected by different incorporation practices. However, little information is known about the microbial mechanisms driving the response of N priming to co-operation of Chinese milk vetch (CMV, Astragalus sinicus L.) and different rates of chemical fertilizers in paddy soils in South China. Here, an anaerobic incubation experiment was conducted to study N priming effects (PE) and their relationships with soil microbial functional genes after CMV incorporation alone (M), co-incorporation of CMV with 100% (normal dosage) chemical fertilizers (MC100), and co-incorporation of CMV with 80% chemical fertilizers (MC80). Co-incorporation of CMV and chemical fertilizers enhanced the short-time scale (the first 20 d of incubation) positive PE of N, while no significant differences existed among the three treatments on day 60 or 90 of incubation (P > 0.05). Compared with the M treatment, gross priming effect (GPE) in the MC100 and MC80 treatments significantly increased by 34.0% and 31.3%, respectively, and net priming effect (NPE) increased by 47.7% and 47.8%, respectively, during the first 20 d of incubation (P < 0.05). This was likely attributed to soil nutrient availability and added substrate quality. The MC100 and MC80 treatments increased the gdhA gene abundance by 5.0% and 9.8%, increased the gdh2 gene abundance by 12.7% and 45.7%, and increased the nasB gene abundance by 9.5% and 41.4%, respectively, in comparison with the M treatment on day 20 of incubation. Correlation analyses indicated that soil microbial functional genes involved in N mineralization (gdhA and gdh2), assimilatory nitrate reduction (nasB), and nitrification (amoB) were significantly correlated with N priming under different incorporation practices during the incubation period (P < 0.05). Thus, co-incorporation of CMV and chemical fertilizers can regulate soil microbial community functional gene structure, which may accelerate mineralization and assimilatory nitrate reduction and inhibit nitrification, thereby increasing the short-term positive PE of N in the present study.

In vitro assessment of antibacterial activity in four endodontic sealers against Staphylococcus aureus and Kocuria rhizophila using agar diffusion test.

In this in vitro study, we assessed the antibacterial efficacy of four endodontic sealers-resin AH26, EndoRez, calcium hydroxide (Apexit), and pure zinc oxide-against Enterococcus faecalis. The agar diffusion test was employed to evaluate the antibacterial efficacy of the sealers in vitro, with distilled water serving as a control. The sealers were prepared following the manufacturer's instructions and placed in wells of 50 agar plates, each inoculated with 15 samples of Kocuria rhizophila and Staphylococcus aureus. Inhibition zones were assessed after 72, 120, and 168 hours of anaerobic incubation at 37°C for 196 hours. Kruskal-Wallis and Friedman tests were used for data analysis. Positive control plates exhibited bacterial growth in all specified periods. AH26 demonstrated significantly higher antibacterial effectiveness against both bacterium types compared to the other sealers (P<0.01). Pure zinc oxide exhibited moderate antibacterial activity, while Apexit and EndoRez showed the lowest activity against S. aureus and no activity against K. rhizophila. AH26 had the highest antibacterial effect, and EndoRez had the lowest (P<0.05). In terms of inhibiting bacterial growth, the effectiveness of root canal sealers was ranked as follows: AH26 > Pure Zinc Oxide >Apexit/EndoRez.

Dietary Guar Gum Alters Colonic Microbial Fermentation in Azoxymethane-Treated Rats

To assess the effects of guar gum on colonic microbial fermentation and cancer development, azoxymethane-treated rats were fed a partially hydrolyzed guar or control diet. Anaerobic fecal incubations were conducted at 8-wk intervals, either without added substrate or with cornstarch or hydrolyzed guar as substrates. Short-chain fatty acids in colonic contents and colonic carcinoma areas were measured at 27 wk. Fecal in vitro fermentation rates were higher for guar-fed rats than for control rats [three-way ANOVA (diet, time, in vitro substrates), P = 0.002]. Fecal in vitro butyrate production was greater for guar-fed rats than for control rats after 3–11 weeks of diet treatment (three-way ANOVA, P = 0.027). Butyrate concentrations of colonic contents at 27 wk were higher in guar-fed than in control rats and higher in the cecum than in the post-cecal colon (two-way ANOVA, P = 0.0001). A regression equation predicting colonic carcinoma area (r2 = 0.279) using propionate and butyrate concentrations of the contents of the post-cecal colon showed propionate as a positive predictor (P < 0.001) and butyrate as a negative predictor (P = 0.033). Our results show that patterns of short-chain fatty acid production may affect the results of fiber-carcinogenesis experiments. Dietary addition of hydrolyzed guar is associated with fecal fermentation low in propionate and high in butyrate; short-chain fatty acid concentrations are greater proximally than distally. These results suggest that butyrate protects against colonic neoplasia, whereas propionate enhances it, and demonstrate that colonic microbiota adapt to produce more butyrate if given time and the proper substrate.

Modeling the kinetics of perfluorooctanoic and perfluorooctane sulfonic acid biodegradation by Acidimicrobium sp. Strain A6 during the feammox process

Per- and polyfluoroalkyl substances (PFAS) are emerging contaminants of concern due to their health effects and persistence in the environment. Although perfluoroalkyl acids (PFAAs), such as perfluorooctanoic acid (PFOA) and perfluorooctanesulfonic acid (PFOS) are very difficult to biodegrade because they are completely saturated with fluorine, it has recently been shown that Acidimicrobium sp. A6 (A6), which oxidizes ammonium under iron reducing conditions (Feammox process), can defluorinate PFAAs. A kinetic model was developed and tested in this study using results from previously published laboratory experiments, augmented with results from additional incubations, to couple the Feammox process to PFAS defluorination. The experimental results show higher Feammox activity and PFAS degradation in the A6 enrichment cultures than in the highly enriched A6 cultures. The coupled experimental and modeling results show that the PFAS defluorination rate is proportional to the rate of ammonium oxidation. The ammonium oxidation rate and the defluorination rate increase monotonically, but not linearly, with increasing A6 biomass. Given that different experiments had different level of Feammox activity, the parameters required to simulate the Feammox varied between A6 cultures. Nonetheless, the kinetic model was able to simulate an anaerobic incubation system and show that PFAS defluorination is proportional to the Feammox activity.

Critical Review on the Inclusion of the Measurement of Nitrogen Produced in Short Incubations (Nan) in the Diagnostic Protocol of Nitrogenous Fertility in Cordoba (Argentina)

N is the nutrient that is applied in the greatest quantity to increase crop yields. Having a soil analysis that allows estimating the N needs of the crop would have economic and environmental benefits. The measurement of total C, total N and the content of N-nitrates are part of the analytical protocol that is carried out in pre-sowing for diagnostic purposes. However, the relationship between these measurements and actual N mineralization in the field is only approximate. To improve the estimation of N availability, it is necessary to consider the initial inorganic N and the mineralized N (Nmin) during the growing season. The search for an accurate predictor of Nmin that can be done quickly in the laboratory has sparked interest in measuring N in short anaerobic incubations or Nan. In this article, the soil properties that intervene in the N mineralization process that affect the measurement of Nan are exposed to know the scope: advantages and disadvantages of the inclusion of Nan in the routine protocol for diagnostic purposes for its use as a predictor of mineralized N during the crop cycle.

Influence of environmental conditions on accumulated polyhydroxybutyrate in municipal activated sludge

Poly(3-hydroxybutyrate) (PHB) was accumulated in full-scale municipal waste activated sludge at pilot scale. After accumulation, the fate of the PHB-rich biomass was evaluated over two weeks as a function of initial pH (5.5, 7.0 and 10), and incubation temperature (25, 37 and 55°C), with or without aeration. PHB became consumed under aerobic conditions as expected with first order rate constants in the range of 0.19 to 0.55 d−1. Under anaerobic conditions, up to 63 percent of the PHB became consumed within the first day (initial pH 7, 55°C). Subsequently, with continued anaerobic conditions, the polymer content remained stable in the biomass. Degradation rates were lower for acidic anaerobic incubation conditions at a lower temperature (25°C). Polymer thermal properties were measured in the dried PHB-rich biomass and for the polymer recovered by solvent extraction using dimethyl carbonate. PHB quality changes in dried biomass, indicated by differences in polymer melt enthalpy, correlated to differences in the extent of PHB extractability. Differences in the expressed PHB-in-biomass melt enthalpy that correlated to the polymer extractability suggested that yields of polymer recovery by extraction can be influenced by the state or quality of the polymer generated during downstream processing. Different post-accumulation process biomass management environments were found to influence the polymer quality and can also influence the extraction of non-polymer biomass. An acidic post-accumulation environment resulted in higher melt enthalpies in the biomass and, consequently, higher extraction efficiencies. Overall, acidic environmental conditions were found to be favourable for preserving both quantity and quality after PHB accumulation in activated sludge.

Characterizations of food-derived ellagic acid-Undaria pinnatifida polysaccharides solid dispersion and its benefits on solubility, dispersity and biotransformation of ellagic acid

The current study was aimed to enhance the solubility, dispersibility and biotransformation efficacy of ellagic acid (EA) by preparing food-derived ellagic acid-Undaria pinnatifida polysaccharides solid dispersion (EA/UPP SD). The results demonstrated that the solubility of EA/UPP SD was improved from 0.014 mg/mL to 0.383 mg/mL, and the enhancement was related to converting to a more amorphous state and restraining its self-aggregation during the mechanochemical process. The structure of EA/UPP SDs was mostly maintained by hydrogen bonds and hydrophobic interactions between EA and UPP. Moreover, the result of in vitro anaerobic incubations showed the biotransformation process was improved with EA/UPP SD addition to substrate due to the advance of microbial accessibility in EA dispersion. Altogether, these results indicated that the EA/UPP SDs expanded the application of EA by increasing the solubility and dispersity, and provided a theoretical basis for bioconversion efficiency enhancement.

Evaluation of hydrocortisone as a strain-dependent growth-regulator of Porphyromonasgingivalis

ObjectivePorphyromonas gingivalis is an oral key pathogen and known to be very diverse in geno- and phenotypes. It is a fastidious bacterium with low O2-tolerance and 3–7 days of incubation are necessary. With growing interest in the field of microbial endocrinology we explored the potential growth-stimulating effect of hydrocortisone (HC, synonym cortisol) on P. gingivalis cultures. Material and methodsSix different P. gingivalis strains were pre-incubated in supplemented Brain-Heart-Infusion broth under appropriate conditions for 24 h, diluted and transferred into microplates. A newly developed and semi-automated spectrophotometric measurement in triplicate, applying a SpectraMax i3x microplate reader at an optical density of 600 nm, was conducted to test growth differences between test group (exposed to a supplement of either 1.25, 2.5, 5, 10, or 20 μg/ml of hydrocortisone) and control group over 48 h of anaerobic incubation (O2 ≤ 1%). Furthermore, strains were also incubated on HC-supplemented blood agar to test for a possible growth-stimulating effect on solid media. ResultsHC significantly stimulated the lag-phase growth of four out of six P. gingivalis strains. Our data suggest a concentration-dependent growth stimulatory effect of HC between 2.5 and 5 μg/ml, while below 1.25 μg/ml and above 10 μg/ml HC either did not stimulate or inhibited growth. ConclusionsHC could reduce the incubation time when isolating P. gingivalis from clinical samples and could boost low biomass cultivations especially during their lag-phase. The growth-modulating effect might be via modulation of virulence factors/quorum sensing gene expression or by reactive oxygen species(ROS)-capturing during early stages of bacterial growth. Further experiments are necessary to explain the mechanism behind our observations.

Enhanced dissolution of arsenic in anaerobic soils upon organic amendment application: acid detergent-soluble organic matter as a potential indicator

Application of organic amendments (OAMs) often enhances arsenic (As) dissolution in paddy soils. Therefore, understanding the properties of OAMs that determine the extent of As dissolution is essential for appropriate soil management. Since As dissolution increases with decrease in soil redox potential caused by microbial respiration, the decomposability of OAMs might be a critical factor controlling As dissolution in amended soils. We hypothesized that contents of acid detergent-soluble organic matter (ADSOM, mainly composed of non-fiber organic matter and hemicellulose) in OAMs can help estimate the potential of OAMs in accelerating As dissolution in soils with added OAMs. Therefore, two contrasting soil types, Andosol and Fluvisol, were mixed with 24 different OAMs and subjected to anaerobic incubation for 14 weeks. Changes in soil Eh and dissolved As contents were monitored throughout the incubation period, and As species in solid phases and ferrous iron (Fe(II)) contents in soils were measured after 2 and 6 weeks of incubation. The higher the ADSOM content in soils with OAMs, the higher the dissolved As contents in soils and the lower the Eh values. Dissolved As also positively correlated with the proportion of As(III) in solid phases and Fe(II) content after 2 and 6 weeks of incubation, indicating that decomposition of ADSOM led to reducing soil conditions, thereby promoting the reduction of As(V) and As-bearing Fe oxides and subsequent As dissolution. The results were consistent between the two types of soils, despite dissolved As content in the Andosol being two orders lower than that in Fluvisol. This is the first study to demonstrate that ADSOM can be a prominent indicator of the potential of OAMs, for promoting As dissolution, when applied to paddy soils.

Effects in air-exposed corn silage of medium chain fatty acids on select spoilage microbes, zoonotic pathogens, and in vitro rumen fermentation

Medium chain fatty acid (MCFA) treatment (0.75% C6, hexanoic; C8, octanoic; C10, decanoic; or equal proportion mixtures of C6:C8:C10:C12 or C8:C10/g; C12 = dodecanoic acid) of aerobically-exposed corn silage on spoilage and pathogenic microbes and rumen fermentation were evaluated in vitro. After 24 h aerobic incubation (37 °C), microbial enumeration revealed 3 log10 colony-forming units (CFU)/g fewer (P = 0.03) wild-type yeast and molds in C8:C10-treated silage than controls. Compared with controls, wild-type enterococci decreased (P < 0.01) in all treatments except the C6:C8:C10:C12 mixture; lactic acid bacteria were decreased (P < 0.01) in all treatments except C6 and the C6:C8:C10:C12 mixture. Total aerobes and inoculated Staphylococcus aureus or Listeria monocytogenes were unaffected by treatment (P > 0.05). Anaerobic incubation (24 h at 39 °C) of ruminal fluid (10 mL) with 0.02 g overnight air-exposed MCFA-treated corn silage revealed higher hydrogen accumulations (P = 0.03) with the C8:C10 mixture than controls. Methane, acetate, propionate, butyrate, or estimates of fermented hexose were unaffected. Acetate:propionate ratios were higher (P < 0.01) and fermentation efficiencies were marginally lower (P < 0.01) with C8- or C8:C10-treated silage than controls. Further research is warranted to optimize treatments to target unwanted microbes without adversely affecting beneficial microbes.

Anaerobic carbon oxidation in sediment of two Brazilian mangrove forests: the influence of tree roots and crab burrows

This study evaluated the effects of biogenic structures (tree roots and crab burrows) on sediment carbon (C), sulfur (S), and iron (Fe) biogeochemistry during the wet season in the Olaria mangrove forests near the city of Cananéia, São Paulo state, Brazil and the Nobrega mangrove forest approximately 2 km from the city. Anaerobic C oxidation pathways were assessed from sediment profiles and anaerobic incubations and related to the abundance of biogenic structures in the form of pneumatophores and crab burrows. Porewater depth profiles of dissolved inorganic carbon (DIC) and SO42- were less steep in the presence than absence of biogenic structures. While Fe(II) appeared unaffected by biogenic structures, Fe(III) levels were significantly higher in the upper 4 cm of the sediment in the presence than absence of vegetation and bioturbation. Surprisingly, the concentration of Fe(III) in this layer was 2-6 times higher in the Nobrega forest (6-13 µmol cm-3) than in the Olaria forest (1.5-6.5 µmol cm-3). Accordingly, depth integrated sulfate reduction (SR) tended to be highest at Olaria, while iron reduction (FeR) was highest at Nobrega. SR accounted for 54-83% of DIC production, with no diference between forested sites, while FeR accounted for 8-24% of DIC production, with a 2-3 times higher contribution in the Nobrega versus the Olaria forest. The results suggest that mangrove roots and crab burrows in mangrove sediments only promote FeR at the expense of SR in the Nobrega forest. It appears that anthropogenic discharge from Cananéia city may have overridden the impact of biogenic structures on sediment redox conditions at Olaria, thereby diminishing the role of FeR without strong stimulation of overall C oxidation rates.

Effects of pyrogenic carbon addition after fire on soil carbon mineralization in the Great Khingan Mountains peatlands (Northeast China)

Wildfires play a critical role in regulating soil carbon (C) budgets in peatland ecosystems, and their frequency and intensity are increasing owing to climate change and human activities. Wildfires not only emit CO2 during the combustion process but also produce pyrogenic carbon (PyC), which accumulates in the soil C pool and influences soil C decomposition. However, the role of PyC after a fire in peatland soil C mineralization has rarely been examined. This study investigated the effects of PyC addition on peatland soil C mineralization and its potential driving mechanisms using an anaerobic/aerobic incubation experiment with peat soils collected from typical peatlands in the Great Khingan Mountains, Northeast China. The effect of PyC was more pronounced under aerobic conditions than under anaerobic conditions. The mean C- mineralization rates of soil were significantly increased by 45.2 ± 15.5 % and 87.6 ± 14.3 % with 10 % PyC250°C addition after the initial stage (D7) of aerobic and anaerobic incubation, but PyC600°C addition caused a to decrease. Compared with PyC600°C, PyC250°C addition significantly increased the available N content and altered the soil microbial activities, which may be the primary reason for the increase in C mineralization rates. Furthermore, adding a high concentration of PyC (10 %) reduced the concentration of phenolics but increased phenol oxidase activity, which promoted C mineralization rates. Thus, PyC250°C addition to peat soils mainly influences the microbial biomass C content through the accumulation of available N and phenolics, which ultimately positively affects C mineralization rates.

Methane and carbon dioxide emissions from the forest floor of a black spruce forest on permafrost in interior Alaska

High-latitude warming has stimulated CO2 and CH4 emissions from permafrost peatland. This study evaluated growing season CH4 and CO2 emissions from a forest floor of a lowland black spruce forest on permafrost in interior Alaska using automated-closed chambers, anaerobic incubation of peat soils and next-generation sequencing. The CH4 emissions from May to October were 723 ± 432 mg C m−2 and 829 ± 628 mg C m−2 from the two Carex wet plots, 124 ± 76 mg C m−2 from the Sphagnum moss plot, and 11 mg C m−2 from the lichen plot (± denotes standard deviation of different years). The CH4 emissions in the peak season showed diurnal variations with a single peak around noon or double peaks depending on the plots. The CH4 emissions were an order of magnitude lower than those for other northern wetlands, possibly because deep, cold peats regulated CH4 production and oxidation at the dry aerobic surface layer. Low water table positions strongly inhibited CH4 emissions in 2018, suggesting the importance of dissolved CH4 pool at the bottom of the active layer. The future trajectory of CH4 emissions could substantially change with water conditions, deep soil temperatures, and sedge compositions.

Influence of Aerobic and Anaerobic Moist Incubation on Selected Nonvolatile Constituents─Comparison to Traditionally Fermented Cocoa Beans

Recently, moist incubation has been proposed as an alternative postharvest processing method for cocoa beans. During this treatment, unfermented and dried cocoa nibs are rehydrated with a lactic acid solution containing ethanol and subsequently incubated for 72 h at 45 °C before drying. Previous studies focused on the aroma formation during this treatment and the further processing of chocolate. The current study focused on the influence of aerobic and anaerobic moist incubation on selected nonvolatile components in comparison with the unfermented raw material and traditionally fermented cocoa. Total phenolic content and total flavan-3-ol content, contents of (+)-catechin, (-)-epicatechin, procyanidins B2 and C1, cinnamtannin A2, methylxanthines (theobromine and caffeine), contents of sugars (sucrose, d-glucose, and d-fructose) and free amino acids (17 proteinogenic amino acids) were determined. The fermentation index was also evaluated. The aerobically incubated and fermented cocoa showed low levels of phenolic compounds in comparison to the unfermented cocoa and the anaerobically incubated cocoa. The level of methylxanthines was unaffected by all treatments. The contents of reducing sugars were more than 2-fold higher after both incubation treatments compared to fermentation. The level of free amino acids liberated was highest after anaerobic incubation followed by fermentation and aerobic incubation. The aerobically incubated cocoa showed the highest FI, while the anaerobically incubated cocoa may be considered under-fermented (FI <1.0). Statistical analysis (ANOVA) showed significant differences between all treatments, which was verified by principal component analysis.

Efficacy of zinc oxide nanoparticles and Bifidobacterium bifidum Extraction on anaerobic bacteria isolated from patients with diarrhea

Infectious acute diarrhea may be prevented with probiotics; because they make up the majority of the colonic flora in breastfed newborns and are likely to contribute to the lower incidence of diarrhea in this population, Bifidobacteria are particularly appealing as probiotics agents. The present study was designed to identify anaerobic bacteria, especially C. difficile the main reason for dysentery associated with antibiotics. Detect the ability of each ZnoNPs and B. bifidum to inhibit bacterial growth. During the period from March to October 2019, (100) children and adults who came to Salah al-deen hospital in Tikrit city participated in the study under the supervision of a physician. All samples were transported using a carry Blair if late one or two hours after collection and culturing. The collected models were also cultured on Xylose Lysine Deoxycholate agar, Salmonella Shigella agar, Eosin methylene blue agar, and MacConkey agar. For initiated diagnoses of the Enterobacteriaceae, Blood agar is used to detect beta-hemolytic isolates, recover enteric bacteria other than Enterobacteriaceae, and evaluate the results of oxidase tests. To diagnose bacterium kinds, biochemical reactions and motility tests were used. Impact of ZnoNPs, and B. bifidum antibiotic In vitro. The results of 100 dysentery feces samples were obtained into (60%) samples for males and (40%) for females. Eighty-two positively impacted anaerobically on growth media like Clostridium complicate agar and MacConkey agar (18%) other than bacteria. In contrast, negative samples revealed 10 (55.56 percent) samples for males and 8 (44.44 percent) samples for females. The same stool samples were taken and cultured on Clostridium difficile agar and MacConkey agar under anaerobic and ideal incubation conditions. 15% and 67% of isolates appeared on MacConkey agar of the total number of samples, while 18% showed negative growth. Finally, Zn NPs showed their ability to inhibit Clostridium complicated segregate lean on the condensation 5 mg/ml, and it caused the inhibitory effect on Clostridium to complicate by 10-22 of the diameter of inhibition. The Inhibition Zone Dimeter ranged from 8 to 25 mm for isolates when condensation was utilized at 2.5 mg/ml. According to the findings, the widths of the inhibitory zones for isolates of C. difficile containing B. bifidum supernatant mg/ml ranged from 9 to 24 mm. Keywords: Zinc oxide nanoparticles, Probiotic, Bifidobacterium bifidum, Clostridium difficile