Enhanced lignin degradation by Irpex lacteus through expanded sterilization further improved the fermentation quality and microbial community during the silage preservation process.

Traditional autoclaving, slow degradation rate and preservation of biomass treated by fungi are the main factors restricting biological treatment. In our previous studies, strains with high efficiency and selective lignin degradation ability were obtained. To further solve the limiting factors of biological treatment, this paper proposed a composite treatment technology, which could replace autoclaves for fungal treatment and improve the preservation and utilization of fungal-pretreated straw. The autoclaved and expanded buckwheat straw were, respectively, degraded by Irpex lacteus for 14 days (CIL, EIL), followed by ensiling of raw materials (CK) and biodegraded straw of CIL and EIL samples with Lactobacillus plantarum for different days, respectively (CP, CIP, EIP). An expansion led to lactic acid bacteria, mold, and yeast of the samples below the detection line, and aerobic bacteria was significantly reduced, indicating a positive sterilization effect. Expansion before I. lacteus significantly enhanced lignin selective degradation by about 6%, and the absolute content of natural detergent solute was about 5% higher than that of the CIL. Moreover, EIL decreased pH by producing higher organic acids. The combination treatment created favorable conditions for ensiling. During ensiling, EIP silage produced high lactic acid about 26.83 g/kg DM and the highest acetic acid about 22.35 g/kg DM, and the pH value could be stable at 4.50. Expansion before I. lacteus optimized the microbial community for ensiling, resulting in EIP silage co-dominated by Lactobacillus, Pediococcus and Weissella, whereas only Lactobacillus was always dominant in CP and CIP silage. Clavispora gradually replaced Irpex in EIP silage, which potentially promoted lactic acid bacteria growth and acetic acid production. In vitro gas production (IVGP) in EIL was increased by 30% relative to CK and was higher than 24% in CIL. The role of expansion was more significant after ensiling, the IVGP in EIP was increased by 22% relative to CP, while that in CIP silage was only increased by 9%. Silage of fungal-treated samples reduced methane emissions by 28% to 31%. The study demonstrated that expansion provides advantages for fungal colonization and delignification, and further improves the microbial community and fermentation quality for silage, enhancing the nutrition and utilization value. This has practical application value for scaling up biological treatment and preserving the fungal-treated lignocellulose.

Immobilization of purified enzyme EreB in metalorganic framework (MOF) mesopores for erythromycin degradation.

Erythromycin, a commonly used macrolide antibiotic, plays a crucial role in both human medicine and animal husbandry. However, its abuse has led to residual presence in the environment, with problems such as the emergence of resistant bacteria and enrichment of resistance genes. These issues pose significant risks to human health. Thus far, there are no effective, environmentally friendly methods to manage this problem. Enzymes can specifically degrade erythromycin without causing other problems, but their unrecyclability and environmental vulnerability hinder large-scale application. Enzyme immobilization may help to solve these problems. This study used Cu-BTC, a synthetic metal-organic framework, to immobilize the erythromycin-degrading enzyme EreB. The loading temperature and enzyme quantity were optimized. The Cu-BTC and EreB@Cu-BTC were characterized by various methods to confirm the preparation of Cu-BTC and immobilization of EreB. The maximum enzyme loading capacity was 66.5 mg g-1. In terms of enzymatic properties, immobilized EreB had improved heat (25-45 °C) and alkaline (6.5-10) tolerance, along with greater affinity between the enzyme and its substrate; Km decreased from 438.49 to 372.30 mM. Recycling was also achieved; after 10 cycles, 57.12% of the enzyme activity was maintained. After composite degradation, the antibacterial activity of erythromycin-containing wastewater was examined; the results showed that the novel composite could completely inactivate erythromycin. In summary, Cu-BTC was an ideal carrier for immobilization of the enzyme EreB, and the EreB@Cu-BTC composite has good prospects for the treatment of erythromycin-containing wastewater.

Effects of Cold Plasma Pretreatment on the Synthesis of Polysaccharide from Pleurotus ostreatus.

Fungal polysaccharides have attracted wide attention because of their medical pharmaceutical and health care value. So far, many efforts have been made in strain improvement to produce polysaccharides on a large scale at low cost. Here, a novel cold plasma-induced strain improvement technology was employed to pretreat Pleurotus ostreatus CGMCC 5.374 by radio-frequency (RF) low-vacuum cold plasma (LVCP) for the purpose of obtaining a high-yield polysaccharide strain. The optimum pretreatment conditions including discharge power, treatment time, and working pressure were determined by single factor and orthogonal experiment in succession. Furthermore, transcriptome analysis was conducted to study the effects of RF-LVCP on cell metabolism and proliferation. Results showed that under the optimal condition of discharge power of 130 W, treatment time of 25 s and working pressure of 140 Pa, polysaccharide content in mycelium was increased by 3.16% after 6 days in comparison to the original strain. Transcriptome analysis showed that RF-LVCP is helpful for specific gene transcription profiles, Gene Ontology (GO) and KEGG pathways, of which the differentially expressed genes (DEGs) were mainly involve with the up-regulation of polysaccharide transport, physiology, synthesis and metabolism, as well as the down-regulation of polysaccharide hydrolysis and macromolecular degradation.

Environmental factors induced macrolide resistance genes in composts consisting of erythromycin fermentation residue, cattle manure, and maize straw.

With the growing concerns about antibiotic resistance, it is more and more important to prevent the environmental pollution caused by antibiotic fermentation residues. In this study, composted erythromycin fermentation residue (EFR) with the mixture of cattle manure and maize straw at ratios of 0:10 (CK), 1:10 (T1), and 3:10 (T2) explores the effects on physicochemical characteristics, mobile genetic elements (MGEs), and antibiotic resistance genes (ARGs). Results reflected that the addition of EFR reduced the carbon/nitrogen ratio of each compost and improved the piles' temperature, which promoted the composting process. However, the contents of Na+, SO42-, and erythromycin were also significantly increased. After 30 days of composting, the degradation rates of erythromycin in CK, T1, and T2 were 72.7%, 20.3%, and 37.1%, respectively. Meanwhile, the total positive rates for 26 detected ARGs in T1 and T2 were 65.4%, whereas that of CK was only 23.1%. Further analysis revealed that ARGs responsible for ribosomal protection, such as ermF, ermT, and erm(35), dominated the composts of T1 and T2, and most were correlated with IS613, electrical conductivity (EC), nitrogen, and Zn2+. Above all, adding EFR helps to improve the nutritional value of composts, but the risks in soil salinization and ARG enrichment caused by high EC and erythromycin content should be further investigated and eliminated.

Biodegradation efficiency and mechanism of erythromycin degradation by Paracoccus versutus W7.

Continuous and excessive usage of erythromycin results in serious environmental pollution and presents a health risk to humans. Biological treatment is considered as an efficient and economical method to remove it from the environment. In this study, a novel erythromycin-degrading bacterial strain, W7, isolated from sewage sludge was identified as Paracoccus versutus. Strain W7 degraded 58.5% of 50 mg/L erythromycin in 72 h under the optimal conditions of 35 °C, pH 7.0, and 0.1% sodium citrate with yeast powder in mineral salt medium. It completely eliminated erythromycin from erythromycin fermentation residue at concentrations of 100 and 300 mg/L within 36 and 60 h, respectively. Erythromycin esterase (EreA) was found to be involved in erythromycin metabolism in this strain and was expressed successfully. EreA could hydrolyze erythromycin, and its maximum activity occurred at pH 8.5 and 35 °C. Finally, six intermediates of erythromycin degraded by strain W7 were detected by high performance liquid chromatography mass spectrometry. Based on the novel intermediates and enzymes, we determined two possible pathways of erythromycin degradation by strain W7. This study broadened our understanding of the erythromycin catabolic processes of P. versutus and developed a feasible microbial strategy for removing erythromycin from erythromycin fermentation residue, wastewater, and other erythromycin-contaminated environments.

Ensiling Improved the Colonization and Degradation Ability of Irpex lacteus in Wheat Straw.

To develop a non-thermal method to replace steam autoclaving for white-rot fungi fermentation, Irpex lacteus spawn was inoculated in wheat straw (WSI) or ensiled WS (WSI) at varying ratios of 10%, 20%, 30%, 40%, and 50%, and incubated at 28 °C for 28 days to determine the effects of the ensiling and inoculation ratio on the colonization and degradation ability of Irpex lacteus in wheat straw (WS). The results demonstrate that ensiling effectively inhibited the growth of aerobic bacteria and molds, as well as other harmful microorganisms in WS, which created a favorable condition for the growth of I. lacteus. After the treatment of I. lacteus, the pH of EWSI decreased to below 5, while that of WSI, except for the feedstocks of WSI-50%, was around 7, indicating that I. lacteus colonized well in the ensiled WS because the substrates dominated by I. lacteus are generally acidic. Correspondingly, except for the molds in WSI-50% samples, the counts of other microorganisms in WSI, such as aerobic bacteria and molds, were significantly higher than those in EWSI (p < 0.05), indicating that contaminant microorganisms had a competitive advantage in non-ensiled substrates. Incubation with I. lacteus did not significantly affect the cellulose content of all samples. However, the NDS content of EWSI was significantly higher than that of WSI (p < 0.05), and the hemicellulose and lignin contents were significantly lower than the latter (p < 0.05), except for the NDS and hemicellulose contents of WSI-50% samples. Correlation analysis revealed a stronger negative correlation between NDS content and the contents of hemicellulose, cellulose, and lignin in EWSI, which could be caused by the destruction of lignin and hemicellulose and the conversion from structural carbohydrates to fungal polysaccharides or other compounds in NDS form. Even for WSI-50% samples, the sugar yield of WS treated with I. lacteus improved with an increasing inoculation ratio, but the ratio was not higher than that of the raw material. However, the sugar yield of EWSI increased by 51-80%, primarily owing to the degradation of lignin and hemicellulose. Above all, ensiling improves the colonization ability of I. lacteus in WS, which promotes the degradation of lignin and hemicellulose and the enzymic hydrolysis of cellulose, so combining ensiling and I. lacteus fermentation has promising potential in the pretreatment of WS.

Effects of pH and Metal Ions on the Hydrothermal Treatment of Penicillin: Kinetic, Pathway, and Antibacterial Activity.

Antibiotic residues lead to the risk of resistance gene enrichment, which is the main reason why penicillin mycelial dreg (PMD) is defined as hazardous waste. Hydrothermal treatment (HT) is an effective method to treat penicillin mycelial dreg, but the degradation mechanism of penicillin is unclear. In the study, we researched the effects of pH (4-10) at 80-100 °C and metal ions (Mn2+, Fe2+, Cu2+, and Zn2+) at several concentrations on the HT of penicillin, identified the degradation products (DPs) under different conditions, and evaluated the antibacterial activity of hydrothermally treated samples. The results show that penicillin degradation kinetics highly consistent with pseudo-first-order model (R2 = 0.9447-0.9999). The degradation rates (k) at pH = 4, 7, and 10 were 0.1603, 0.0039, and 0.0485 min-1, indicating acidic conditions were more conducive to penicillin degradation. Among the four tested metal ions, Zn2+ had the most significant catalytic effect. Adding 5 mg·L-1 Zn2+ caused 100% degradation rate at pH = 7 after HT for 60 min. Six degradation products (DPs) with low mass-to-charge (m/z ≤ 335) were detected under acidic condition. However, only two and three DPs were observed in the samples catalyzed by Zn2+ and alkali, respectively, and penilloic acid (m/z = 309) was the main DPs under these conditions. Furthermore, no antibacterial activity to Bacillus pumilus was detected in the medium with up to 50% addition of the treated samples under acidic condition. Even though acid, alkali, and some metal ions can improve the degradation ability of penicillin, it was found that the most effective way for removing its anti-bacterial activity was under the acidic condition. Therefore, resistance residue indicates the amount of additive in the process of resource utilization, and avoids the enrichment of resistance genes.

Immobilization of EreB on Acid-Modified Palygorskite for Highly Efficient Degradation of Erythromycin.

Erythromycin is one of the most commonly used macrolide antibiotics. However, its pollution of the ecosystem is a significant risk to human health worldwide. Currently, there are no effective and environmentally friendly methods to resolve this issue. Although erythromycin esterase B (EreB) specifically degrades erythromycin, its non-recyclability and fragility limit the large-scale application of this enzyme. In this work, palygorskite was selected as a carrier for enzyme immobilization. The enzyme was attached to palygorskite via a crosslinking reaction to construct an effective erythromycin-degradation material (i.e., EreB@modified palygorskite), which was characterized using FT-IR, SEM, XRD, and Brunauer-Emmett-Teller techniques. The results suggested the successful modification of the material and the loading of the enzyme. The immobilized enzyme had a higher stability over varying temperatures (25-65 °C) and pH values (6.5-10.0) than the free enzyme, and the maximum rate of reaction (Vmax) and the turnover number (kcat) of the enzyme increased to 0.01 mM min-1 and 169 min-1, respectively, according to the enzyme-kinetics measurements. The EreB@modified palygorskite maintained about 45% of its activity after 10 cycles, and degraded erythromycin in polluted water to 20 mg L-1 within 300 min. These results indicate that EreB could serve as an effective immobilizing carrier for erythromycin degradation at the industrial scale.

Evolution of antibiotic resistance genes and bacterial community during erythromycin fermentation residue composting.

The removal efficiency of antibiotic resistance genes (ARGs) is the biggest challenge for the treatment of erythromycin fermentation residue (EFR). In the current research, 0% (control), 10% (T1), and 30% (T2) spray-dried EFR were composted with bulking materials, consisting of cattle manure and maize straw, for 30 days. Environmental factors and bacterial community on the behaviors of ARGs were further investigated. Apart from the high levels of erythromycin, the electrical conductivities were also increased by 66.7% and 291.7% in the samples of T1 and T2, respectively. After 30 days of composting, total ARGs in the samples of control were decreased by 78.1%-91.2%, but those of T1 and T2 were increased 14.5-16.7- and 38.5-68.7-fold. ARGs related to ribosomal protection (erm) dominated the samples of T1 and T2 at D 13 and 30, especially that ermF accounted for more than 80% of the total ARGs. Furthermore, the results of bacterial community revealed that EFR promoted the growth of Proteobacteria and Bacteroidetes, but inhibited that of Actinobacteria, Verrucomicrobia and Chloroflexi. Network analysis revealed that the enriched ARGs had strong correlation with seven bacterial genera, including Halomonas, Oceanobacillus, and Alcaligenes, most of which are halotolerant. Above all, erythromycin combined with high salinity can have synergistic effect on the enrichment of ARGs and their hosts.

[Advances in biodegradation of macrolide antibiotics].

Macrolide antibiotics are a class of broad-spectrum antibiotics with the macrolide as core nucleus. Recently, antibiotic pollution has become an important environmental problem due to the irregular production and abuse of macrolide antibiotics. Microbial degradation is one of the most effective methods to deal with antibiotic pollution. This review summarizes the current status of environmental pollution caused by macrolide antibiotics, the degradation strains, the degradation enzymes, the degradation pathways and the microbial processes for degrading macrolide antibiotics. Moreover, the critical challenges on the biodegradation of macrolide antibiotics were also discussed.

Safety of composts consisting of hydrothermally treated penicillin fermentation residue: Degradation products, antibiotic resistance genes and bacterial diversity.

Combining hydrothermal treatment and composting is an effective method to dispose of penicillin fermentation residue (PFR), but the safety and related mechanism are still unclear. In this study, penicillin solution was hydrothermally treated to decipher its degradation mechanism, and then hydrothermally treated PFR (HT-PFR) was mixed with bulking agents at ratios of 2:0 (CK), 2:1.5 (T1), and 2:5 (T2) to determine the absolute abundance of antibiotic resistance genes (ARGs) and the succession of bacterial community. Results showed that penicillin was degraded to several new compounds without the initial lactam structure after hydrothermal treatment. During composting, temperature and pH of the composts increased with the raising of HT-PFR proportion, except the pH at days 2. After 52 days of composting, the absolute copies of ARGs (blaTEM, blaCMY2, and blaSFO) and the relative abundance of bacteria related to pathogens were reduced significantly (P < 0.05). Especially, the total amount of ARGs in the samples of CK and T1 were decreased to equal level (around 5 log10 copies/g), which indicated that more ARGs were degraded in the latter by the composting process. In the CK samples, Bacteroidetes and Proteobacteria accounted for ~69.8% of the total bacteria, but they were gradually replaced by Firmicutes with increasing proportions of HT-PFR, which can be caused by the high protein content in PFR. Consisting with bacterial community, more gram-positive bacteria were observed in T1 and T2, and most of them are related to manganese oxidation and chitinolysis. As composting proceeded, bacteria having symbiotic or pathogenic relationships with animals and plants were reduced, but those related to ureolysis and cellulolysis were enriched. Above all, hydrothermal treatment is effective in destroying the lactam structure of penicillin, which makes that most ARGs and pathogenic bacteria are eliminated in the subsequent composting.

Effects of added thermally treated penicillin fermentation residues on the quality and safety of composts.

Thermal treatment and composting are effective methods of degrading antibiotics and organic matter in penicillin fermentation residues (PFR), respectively. However, the composting efficiency and environmental safety of thermally treated PFR (HT-PFR) remain unclear. In this study, HT-PFR was composted with cattle manure and maize straw at ratios of 0:1:1 (CK), 1.5:1:1 (T1), and 5:1:1 (T2). Changes in physicochemical properties, seed germination index (GI), and microbial antibiotic resistance genes (ARGs) were determined. Addition of HT-PFR significantly reduced the C:N ratio of each compost (P < 0.05) and prolonged the thermophilic stage. The GI of CK and T1 composts increased during processing, whereas that of T2 compost remained low. The PO43- concentrations of T1 and T2 composts were 6.3- and 11.1-fold higher than that of CK, respectively. HT-PFR contained relatively small amounts of mineral elements, and composting it with cattle manure and maize straw provided balanced nutrients for plant growth. After 52 days of composting, most ARGs of the microflora were reduced to low levels, but blaTEM increased significantly in T2 compost. Overall, composting HT-PFR at up to 42% of a mix containing equal parts of cattle manure and wheat straw is an environmentally safe and effective way of transforming it into organic fertilizer.

Isolation and identification of a novel erythromycin-degrading fungus, Curvularia sp. RJJ-5, and its degradation pathway.

Erythromycin pollution is an important risk to the ecosystem and human health worldwide. Thus, it is urgent to develop effective approaches to decontaminate erythromycin. In this study, we successfully isolated a novel erythromycin-degrading fungus from an erythromycin-contaminated site. The erythromycin biodegradation characteristics were investigated in mineral salt medium with erythromycin as the sole carbon and energy source. The metabolites of erythromycin degraded by fungus were identified and used to derive the degradation pathway. Based on morphological and phylogenetic analyses, the isolated strain was named Curvularia sp. RJJ-5 (MN759651). Optimal degradation conditions for strain RJJ-5 were 30°C, and pH 6.0 with 100 mg L-1 erythromycin substrate. The strain could degrade 75.69% erythromycin under this condition. The following metabolites were detected: 3-depyranosyloxy erythromycin A, 7,12-dyhydroxy-6-deoxyerythronolide B, 2,4,6,8,10,12-hexamethyl-3,5,6,11,12,13-hexahydroxy-9-ketopentadecanoic acid and cladinose. It was deduced that the erythromycin A was degraded to 3-depyranosyloxy erythromycin A by glycoside hydrolase in the initial reaction. These results imply that Curvularia sp. RJJ-5 is a novel erythromycin-degrading fungus that can hydrolyze erythromycin using a glycoside hydrolase and has great potential for removing erythromycin from mycelial dreg and the contaminated environment.

Biodegradation of erythromycin by Delftia lacustris RJJ-61 and characterization of its erythromycin esterase.

The residual erythromycin in fermentation waste can pollute the environment and threaten human health. However, there are no effective approaches to remedy this issue. In this study, an erythromycin-degrading bacterium named RJJ-61 was isolated and identified as a strain of Delftia lacustris based on morphological and phylogenetic analyses. The degradation ability of this strain was also evaluated; it could degrade 45.18% of erythromycin at 35°C in 120 h. Furthermore, the key degradation gene ereA was cloned from strain RJJ-61 and expressed in Escherichia coli BL21; the molecular weight of the expressed protein was ~45 kDa. The enzyme activity of EreA was 108.0 mU ml-1 at 35°C and pH 7.0. Finally, the EreA protein was used to degrade erythromycin from mycelial dregs and 50% diluted solution, and the removal rates in them were 41.42% and 69.78%, respectively. In summary, D. lacustris RJJ-61 is a novel erythromycin-degrading strain that has great potential to remove erythromycin pollutants from the environment.

Effects of Ensiling Oxytropis glabra with Whole-Plant Corn at Different Proportions on Fermentation Quality, Alkaloid Swainsonine Content, and Lactic Acid Bacteria Populations.

Oxytropis glabra (OG) is a leguminous forage that is potentially valuable for solving the shortage of feed for livestock production, while, in large quantities, it may be toxic because of its swainsonine (SW) content. In this study, OG was ensiled with whole-plant corn (Zea mays L.) at 10:0, 9:1, 8:2, 7:3, 6:4, 5:5, 4:6, and 0:10 ratios on a fresh matter basis, and, after 60 d of ensiling, the chemical composition, fermentation characteristic, SW removal rate, lactic acid bacteria (LAB) populations, and their capabilities for SW removal were analyzed. As the proportion of corn in the silage increased, the pH, as well as the propionic acid, ammonia-N, dry matter, crude protein, and SW contents, decreased linearly, while the lactic acid, neutral detergent fiber, and residual water-soluble carbohydrate contents increased linearly. Lactobacillus plantarum was the most common microorganism present in all mixture silages. Lactobacillus amylovorus and Lactobacillusbrevis were prevalent at lower ratios of corn to OG. Meanwhile, the LAB strains belong to L. amylovorus and L. plantarum had a higher SW removal rate. Our results suggested that ensiling OG with whole-plant corn improves fermentation and decreases SW content, and that 5:5 is the optimal ratio, so this type of mixed silage could make OG useable for ruminant production.

Biodegradation of acephate by Bacillus paramycoides NDZ and its degradation pathway.

Acephate is widely used in agriculture, but its poisonous metabolites and poor sorption characteristics make it a serious environmental pollutant and toxicant to human health. To screen novel bacteria for biodegradation of acephate and uncover its degradation pathway, a strain called NDZ that is capable of utilizing acephate as a sole carbon and energy source was isolated from severely contaminated cultivated land. The bacterium was identified as Bacillus paramycoides based on 16S rDNA sequence analyses. The growth and degradation capacities of B. paramycoides NDZ under different conditions were studied using optical density at 600 nm (OD600) and high-performance liquid chromatography (HPLC). The results showed that B. paramycoides NDZ can grow well with acephate as its sole carbon source (OD600 = 0.76), and degraded about 76% of acephate in mineral salt medium with an initial concentration of 500 mg/L within 48 h. The results of response surface methodology revealed the optimal conditions for degradation was 36 ℃ and pH 6.85 with 526 mg/L acephate. Gas chromatography-mass spectrometry showed that methamidophos was the main metabolite of B. paramycoides NDZ, different from the degradation products of high-temperature steam (121 °C, 103 kPa). Based on the detection of this intermediate, we inferred that acephate was degraded to methamidophos through hydrolysis of the amide linkage, after which methamidophos was degraded to some small molecules, which can be metabolized easily by the bacterium. In summary, B. paramycoides NDZ is a potentially useful bacterium for acephate degradation and remediation of contaminated soils.

Vitamin A and E in the total mixed ration as influenced by ensiling and the type of herbage.

The objective of this study was to evaluate the effects of ensiling on vitamin A (retinol) and vitamin E (α-tocopherol) contents in the total mixed ration (TMR) containing different types of herbage. Oat hay (O-TMR), alfalfa hay (A-TMR) and oat hay + alfalfa hay (OA-TMR) were separately mixed with soybean milk residue, corn meal, soybean meal, salt and a vitamin-mineral supplement to make the TMR. The TMR was sampled after 0, 7, 14, 28 and 56 days of ensiling. The fermentation quality, chemical composition and contents of vitamins A and E were determined. The vitamin A content was affected by the ensiling and herbage type (p < 0.05). After 56 days of ensiling, the three TMR silages had good fermentation quality, but the vitamin A content of O-TMR, OA-TMR and A-TMR decreased by 59.4%, 58.1% and 53.7%, respectively. Moreover, the content of vitamin A was positively correlated with the pH and negatively correlated with the lactic acid content during the 56 days of ensiling of the TMR silages. However, there were no effects of ensiling and herbage type on the vitamin E content. Thus, the preservation strategy for vitamin A in the TMR during ensiling requires further study.

Yeast population dynamics on air exposure in total mixed ration silage with sweet potato residue.

To investigate the yeast population dynamics during air exposure in total mixed ration (TMR) silage containing sweet potato residue. TMR were ensiled in laboratory silos (1 kg) with or without two lactic acid bacteria strains, Lactobacillus plantarum (LP), and Lactobacillus amylovorus (LA). Fermentation characteristics were measured and yeast population was investigated by ITS1 region gene sequencing using Illumina MiSeq platform. All treatments were well ensiled, and L. amylovorus improved aerobic stability. During aerobic exposure, Pichia kudriavzevii was detected with increased relative abundance in all treatments and more relative abundant in LP. Pichia fermentans was more relative abundant in control. Higher relative abundance of Pichia anomala was detected in deteriorating LP. The relative abundance of Pichia ohmeri increased during later aerobic exposure in the control and LA, with a significant increase in the count of yeast population. Despite Cryptococcus was detected more relative abundant during early stage of aerobic exposure, the yeast population was below the detection limit. Aerobic deterioration was characterized by an increase in operational taxonomic units of Pichia. High relative abundance of P. anomala and P. kudriavzevii made aerobic deterioration easier. Inhibition of P. fermentans might be an effective strategy for improving the aerobic stability to some instance.

Isolation and characterization of the novel oil-degrading strain Kosakonia cowanii IUMR B67 and expression of the degradation enzyme.

Oils in food waste can pollute the environment and negatively affect human health. Biodegradation is a promising method for disposing of waste edible oils. In this study, an oil-degrading bacterium was isolated from kitchen waste for efficient degradation of edible oils. Its growth and oil degradation characteristics were investigated in basic salt medium with edible oils as the sole carbon and energy source; the triacylglycerol lipase gene (EC 3.1.1.3) was cloned and expressed in Escherichia coli. A novel oil-degrading bacterium assigned as IUMR B67 was successfully isolated. Morphological and molecular analyses revealed that strain IUMR B67 belongs to Kosakonia cowanii. After 144 h of incubation, the oil degradation rate at 37°C was 95.80%. Optimal conditions for IUMR B67 were recorded at 37°C and 0.1% NaCl with 0.1% ammonium sulfate supplementation. The lipase gene of strain IUMR B67 was determined to be 912 base pairs, and the lipase activity of the expressed protein was 3.02 U/mL, which was significantly higher than the control (P < 0.05). Overall, Kosakonia cowanii IUMR B67 is a novel edible oil-degrading strain that can hydrolyze oil via its lipase activity, which may be useful in the disposal of oils and oily food waste.

Metagenome analyses reveal the role of Clostridium perfringens in alfalfa silage anaerobic deterioration.

The clostridial fermentation caused by the outgrowth of Clostridia was mainly responsible for the silage anaerobic deterioration. Our previous results showed that Clostridium perfringens dominated the clostridial community in poor-fermented alfalfa silage. This study was conducted to further examine the role of C. perfringens in silage anaerobic deterioration through fermentation products and the microbial community analyses. Direct-cut alfalfa was ensiled with C. perfringens contamination (CKC) or with the addition of Lactobacillus plantarum, sucrose and C. perfringens (LSC). Contamination with C. perfringens enhanced the clostridial fermentation in CKC silage, as indicated by high contents of butyric acid, ammonia nitrogen and Clostridia, while LSC silage was well preserved. The genera Bifidobacterium, Garciella and Clostridium dominated the bacterial community in CKC silage, while predominate genus was replaced by Lactobacillus in LSC silage. The clostridial community in CKC silage was dominated by Garciella sp. (26.9 to 58.1%) and C. tyrobutyricum (24.4 to 48.6%), while the relative abundance of C. perfringens was below 5.0%. Therefore, the effect of Clostridia contamination on ensiling fermentation was dependent on the ensilability of the silage material. Garciella sp. and C. tyrobutyricum, rather than C. perfringens, played dominant role in the clostridial fermentation in CKC silage.