Effects of microplastics on functional genes related to CH4 and N2O metabolism in bacteriophages during manure composting and its planting applications.

Microplastics (MPs), as a new type of pollutant, widely exist in livestock and poultry breeding and agricultural soils. However, research on MPs pollution on greenhouse gas emissions in combined planting and breeding systems is lacking, especially from the perspective of phage horizontal gene transfer. Therefore, this paper explores the effects of MPs on functional genes related to CH4 and N2O metabolism in bacteriophages during manure composting and its planting applications. The results of the study indicated that the addition of MPs had an impact on both the physicochemical properties and microbial community structure of manure during the composting process and on the compost-applied rhizosphere soil of lactuca (Lactuca sativa). Specifically, on day 7 of composting, mcrA/pmoA and (nirS+nirK) levels in bacteria in the MP group significantly increased. Additionally, it was observed that the MP group had higher average temperatures during the high-temperature period of composting, which led to a rapid reduction in phages. However, the phage levels quickly recovered during the cooling period. Furthermore, the addition of MPs to the rhizosphere soil resulted in higher levels of nirK. These changes may affect greenhouse gas emissions.

Activity fingerprinting of polysaccharides on oral, gut, pancreas and lung microbiota in diabetic rats.

The modern rise in type 2 diabetes mellitus (T2DM) and its correlation to commensal microbiota have elicited global concern about the patterns of microbial action in the host. With the exception of that linked to gut, microbiota were also colonized in pancreas, oral, and lung, contributing to the physiopathology of T2DM. In this study, we aimed to explore the protective effects of Ganoderma atrum polysaccharide (PSG) and White Hyacinth Bean polysaccharide (WHBP) on the intestine, pancreas, oral, and lung microbiota in T2DM rats. Here we showed that, despite capacities of polysaccharides that exerted similar protective effects on hyperglycemia, dyslipidemia, insulin resistance and dysbacteriosis in T2DM rats, PSG and WHBP were able to be characterized by their own "target" bacteria, which could be proposed for activity-fingerprinting of polysaccharide species. Furthermore, we found a mutual bacteria spectrum in the pancreas and lung, and most bacteria could be tracked to oral or gut samples. Notably, the overlapping areas of the microbiota profile between organs (pancreas, lung) and saliva were more than in the gut, suggesting that a saliva sample was also of interest to serve as a "telltale sign" for judging pancreatic injury. Together, these microbiota interactions provided a new potential to harvest alternative samples for disease surveillance. Meanwhile, polysaccharides had anti-T2DM abilities, which could be distinguished by their own characteristic bacteria.

Cyclic AMP and biofilms reveal the synergistic proliferation strategy of Pseudomonas aeruginosa and Escherichia coli under the costimulation of high concentrations of microplastics and enrofloxacin.

Microplastics (MPs) provide attachment sites for biofilm formation of microorganisms, which can promote their resistance to environmental stress has been proved. However, the effect of MPs on synergy survival among microorganisms under antibiotic stress remains unclear. In the present study, the proliferation of Escherichia coli and Pseudomonas aeruginosa was assessed under enrofloxacin stress with the influence of MPs. Here, MPs reduced the growth speed of E. coli and enhanced that of P. aeruginosa, especially at 12 h, but the final value of OD600 and CFU of both bacteria not be influenced. E. coli was enrofloxacin sensitive (MIC = 0.25 µg/mL), and a high MP concentration in the presence of enrofloxacin notably enhanced the biofilm formation ability of P. aeruginosa, but proliferation decreased. In the coculture system, the proliferation of E. coli (increased 1.42-fold) and P. aeruginosa (increased 1.06-fold) both increased under enrofloxacin stress (0.25 µg/mL) with high-concentration MP addition. P. aeruginosa may provide the biofilm matrix for E. coli to resist the stress of enrofloxacin. The high concentration of cyclic AMP secreted by E. coli may slightly inhibited biofilm formation, leading to a decrease in the fitness cost of P. aeruginosa; thus, the proliferation of P. aeruginosa increased. The present study is the first to show that MP combined with antibiotics stimulates the metabolic cooperation of bacteria to promote proliferation.

A sustainable and economic strategy to reduce risk antibiotic resistance genes during poultry manure bioconversion by black soldier fly Hermetia illucens larvae: Larval density adjustment.

Black soldier fly (Hermetia illucens) larvae (BSFL) are common insects that are known for bioconversion of organic waste into a sustainable utilization resource. However, a strategy to increase antibiotic resistance gene (ARG) elimination in sustainable and economic ways through BSFL is lacking. In the present study, different larval densities were employed to assess the mcr-1 and tetX elimination abilities, and potential mechanisms were investigated. The application and economic value of each larval density were also analyzed. The results showed that the 100 larvae cultured in 100 g of manure group had the best density because the comprehensive disadvantage evaluation ratio was the lowest (14.97%, good bioconversion manure quality, low ARG deposition risk and reasonable larvae input cost). Further investigation showed that mcr-1 could be significantly decreased by BSFL bioconversion (4.42 ×107 copies/g reduced to 4.79 ×106-2.14 ×105 copies/g)(P＜0.05); however, mcr-1 was increasingly deposited in the larval gut with increasing larval density. The tetX abundance was stabilized by BSFL bioconversion, except that the abundance at the lowest larval density increased (1.22 ×1010 copies/g increase, 34-fold). Escherichia was the host of mcr-1 and tetX in all samples, especially in fresh manure; Alcaligenes was the host of tetX in bioconversion manure; and the abundance of Alcaligenes was highly correlated with the pH of bioconversion manure. The pH of bioconversion manure was extremely correlated with the density of larvae. Klebsiella and Providencia were both hosts of tetX in the BSF larval gut, and Providencia was also the host of mcr-1 in the BSF larval gut. The density of larvae influenced the bioconversion manure quality and caused the ARG host abundance to change to control the abundance of ARGs, suggesting that larval density adjustment was a useful strategy to manage the ARG risk during BSFL manure bioconversion.

Breed differences in the expression levels of gga-miR-222a in laying hens influenced H2S production by regulating methionine synthase genes in gut bacteria.

BACKGROUND: The microbiota in the cecum of laying hens is crucial for host digestion, metabolism, and odor gas production. The results of recent studies have suggested that host microRNAs (miRNAs) can regulate gene expression of the gut microbiota. In the present study, the expression profiles of host-derived miRNAs in the cecal content of two laying hen breeds; Hy-line Gray and Lohmann Pink, which have dissimilar H2S production, were characterized; and their effects on H2S production by regulating the expression of gut microbiota-associated genes were demonstrated. RESULTS: The differential expression of microbial serine O-acetyltransferase, methionine synthase, aspartate aminotransferase, methionine-gamma-lyase, and adenylylsulfate kinase between the two hen breeds resulted in lower H2S production in the Hy-line hens. The results also revealed the presence of miRNA exosomes in the cecal content of laying hens, and an analysis of potential miRNA-target relationships between 9 differentially expressed miRNAs and 9 differentially expressed microbial genes related to H2S production identified two methionine synthase genes, Odosp_3416 and BF9343_2953, that are targeted by gga-miR-222a. Interestingly, in vitro fermentation results showed that gga-miR-222a upregulates the expression of these genes, which increased methionine concentrations but decreased H2S production and soluble sulfide concentrations, indicating the potential of host-derived gga-miR-222a to reduce H2S emission in laying hens. CONCLUSION: The findings of the present study reveal both a physiological role by which miRNAs shape the cecal microbiota of laying hens and a strategy to use host miRNAs to manipulate the microbiome and actively express key microbial genes to reduce H2S emissions and breed environmentally friendly laying hens. Video Abstract.

Chimonanthus nitens Oliv. essential oil mitigates lipopolysaccharide-induced acute lung injury in rats.

This study aimed to evaluate the effect of Chimonanthus nitens Oliv. essential oil (named CEO) on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in rats. In the present study, 21 compounds were characterized in CEO by gas chromatography-mass spectrometry analysis. Furthermore, animal data suggested that CEO could protect rats against ALI, as evidence by increasing white blood cell count, reducing immune organ index and improving lung histopathological changes in rats subjected to LPS. Reduction of the levels of IL-1ß was also shown during CEO-triggering lung protection in rats. Meanwhile, these protective effects of CEO were accompanied by the attenuation of lipid oxidation, and elevation of antioxidant enzymes, suggesting that enhancement of antioxidant defense was linked to its lung protection. Moreover, a combination with CEO and LPS significantly elevated short-chain fatty acids (SCFAs) compared with LPS alone via increasing propionic, i-butyric, butyric and i-valeric acid on LPS-induced ALI in rats. Therefore, our findings indicated that CEO could alleviate LPS-caused ALI in rats by controlling aberrant inflammation, correcting the redox system, and modulating SCFAs in rats.

Elimination and analysis of mcr-1 and blaNDM-1 in different composting pile layers under semipermeable membrane composting with copper-contaminated poultry manure.

The mcr-1 and blaNDM-1 elimination in copper contamination poultry manure was evaluated by semi-permeable membrane composting. The results showed the mcr-1 in control and high copper groups could not be removed, but mcr-1 decreased superlatively 80.1% in low copper treatment group. BlaNDM-1 was increased after composting, especially the copper addition groups, the results indicated that the relative abundance of mcr-1 and blaNDM-1 was obviously different in the different pile layers of copper treatment groups. Three mobile gene elements (MEGs) correlated both mcr-1 and blaNDM-1,copB correlated mcr-1, czcA and copA correlated both mcr-1 and blaNDM-1. The major phyla were Firmicutes, Bacteroidota, Actinobacteriota and Proteobacteria in all layers. The correlation analysis showed that the antibiotic resistance genes (ARGs) potential hosts could be influenced by copper form and physicochemical parameters. Semi-permeable membrane composting could decrease the abundance of major potential pathogens. Furthermore, the composting pile was not homogeneous by semi-permeable membrane composting.

Occurrence of microplastic in livestock and poultry manure in South China.

Microplastic (MP) contamination in soil has attracted much attention, and increasing evidence suggests that MPs can accumulate in agricultural soils through fertilization by compost. In addition, the most common raw materials for composting are livestock and poultry manure wastes. Because the presence of MPs may threaten the safe utilization of fertilizers composted by livestock and poultry wastes during crop planting, it is necessary to understand the contamination risk of MPs present in livestock and poultry manure. In this study, the distribution of MPs in 19 livestock and poultry farms with 3 different species was investigated by using FTIR microscopy. A total of 115 items manure MPs and 18 items feed MPs were identified as PP and PE types dominated by colorful fragments and fibers. Furthermore, after comparing the compositions of plastic products used in the feeding process, we proposed two transport pathways for MP pollution in manure and one potential transport pathway in feeds. Our result proved that the application of swine and poultry manure directly could be a new route of MPs in agricultural soil, furthermore, the presence of MPs could threaten the safety of resource utilization in agricultural soil by using swine and poultry manure for manure potentially. Not, only that, our study also provided a reference for the remediation of MP-contaminated soil.

A novel method for extraction of polypropylene microplastics in swine manure.

With the development of modern industry and agriculture, plentiful microplastics (MPs) were produced as a result of the abuse of plastic. The widespread presence of MPs in soils has caused coastal ecological environment pollution. Previous research has shown that fertilizer is one pathway for the entry of MPs into agricultural soils. Meanwhile, livestock manure is a major fertilizer for crops, and the application of livestock manure compost creates a potential pathway for MPs to enter soils. Thus, MPs may exist in livestock manure from the process of livestock breeding and ultimately contaminate agricultural soils. Based on the increasing attention to MP pollution, manure-born MPs will attract more interest in the future. Thus, the present study compares the extraction effects of centrifugation with fractional distillation, and an improved method is introduced to extract polypropylene (PP) from different types of swine manure. The numbers of particles and fibers were determined using a camera (MS60) connected to a stereomicroscope (Mshot MZ62), and the results showed that the recovery rate of plastic particles in swine manure based on different added numbers ranged from 71.43% ± 8.36 to 96.67% ± 3.33 with the centrifugation method, and only 31.11% ± 10.56 to 43.33% ± 12.56 using fractional distilling. The recovery rate for fibers was generally higher than for particles, especially using centrifugation, and ranged from 95.67% ± 1.58 to 100% ± 0, while the rate of fiber recovery using fractional distillation ranged from 39.44% ± 10.66 to 39.44 ± 10.66. The results of recovery rates using the two methods show that the effect of extraction by centrifugation is superior to the method of fractional distillation, with a recovery rate of approximately 100% for fibers and 90% for particles. The recovery number of microplastics evaluated with a line regression model was acceptable. Graphical abstract.

The Combination of Lead and Bacillus coagulans R11 Increased the Concentration of Alpha-Solanine in the Cecum of Laying Hens and the Pathogens Abundance Decreased.

Alpha-solanine is an alkaloid that can inhibit the growth of pathogens and cancer cells, the present study proved that feeding with Bacillus coagulans R11 increases the concentration of alpha-solanine in the cecum of laying hens, which also decreases the abundance of potential pathogens. In addition, the bacteria genera, metabolism pathways and its proteins involved in the biosynthesis of alpha-solanine in the cecum were also characterized. The results showed that B. coagulans R11 feeding could increase the concentration of alpha-solanine, even with lead exposure. Mevalonic acid and MEP/DOXP pathways were both participated in the biosynthesis of alpha-solanine; at the same time, the gut metabolites (S)-2-amino-6-oxohexanoate, N2-succinyl-L-ornithine and the bacteria proteins atoB, ispH were shown to be crucial role in the biosynthesis of alpha-solanine in the gut. The genera Faecalibacterium sp. An77 and Faecalibacterium sp. An58 2 were important in the biosynthesis of alpha-solanine, which provided the key proteins atoB and ispH. In addition, alpha-solanine could decrease the abundance of Prevotella sp. 109 and Prevotella marshii. In conclusion, alpha-solanine could be biosynthesized by cecal microorganisms with the stimulation of B. coagulans R11 in the intestine of laying hens, in addition, alpha-solanine was the main compound which also decreased the abundance of gut potential.

Protective effect of Ganoderma atrum polysaccharides in acute lung injury rats and its metabolomics.

The present study aimed to evaluate effect of Ganoderma atrum polysaccharide (PSG) on acute lung injury (ALI) rats and its mechanisms. Results showed that PSG exhibited protective effects against ALI by maintaining pulmonary histology, reducing levels of pro-inflammatory cytokines and NO both in serum and lung tissue. Moreover, this study further evaluated the metabolic effects of PSG based on UPLC-Triple-TOF/MS metabolomics analysis in rats. Compared with control group, LysoPC (18:2), LPA (18:1), taurocholic acid, L-histidine, and L-tryptophan were identified as metabolic biomarkers in serum of ALI group. Furthermore, biological pathways analysis demonstrated that histidine metabolism, nitrogen metabolism, tryptophan and part glycerophospholipids metabolism were notably modified by PSG treatment in ALI rats. Additionally, improved gut microbial metabolite short-chain fatty acids were found after intake of PSG in ALI rat. Altogether, PSG could control ALI-induced aberrant inflammation and its mechanisms were linked to inhibit release of pro-inflammatory mediators and reverse metabolic pathway disturbances.

Effects of a Ganoderma atrum polysaccharide against pancreatic damage in streptozotocin-induced diabetic mice.

This study aimed at exploring the role of a Ganoderma atrum polysaccharide (PSG-1) in pancreatic damage in streptozotocin (STZ)-induced type 1 diabetes mellitus (T1DM) mice. The results suggested that blood glucose was significantly increased in the STZ group in comparison with the control group. After 4 weeks of treatment with PSG-1 or metformin (MET), blood glucose levels in the PSG-1 and MET groups were apparently lower than in the STZ group, indicating that PSG-1 triggered hypoglycemic effects in vivo. Moreover, experiments demonstrated that PSG-1 markedly decreased apoptosis of islet cells by inhibiting the mitochondrial apoptotic pathway and activating the PI3K/Akt survival pathway. PSG-1 also exerted anti-inflammatory effects, as evidenced by the dramatically decreased levels of IL-1ß, TNF-α and INF-γ and restraint of the TLR4-dependent NF-κB signal pathway. Meanwhile, PSG-1 maintained homeostasis of redox systems by increasing the activities of anti-oxidant enzymes and decreasing the amount of malondialdehyde in the pancreas. Together, these data provide evidence that PSG-1 can be employed as an alternative dietary supplement to ameliorate T1DM.

Ganoderma atrum polysaccharide improves doxorubicin-induced cardiotoxicity in mice by regulation of apoptotic pathway in mitochondria.

The present study aimed to determine the cardioprotective effect of Ganoderma atrum polysaccharide (PSG-1) in doxorubicin (DOX)-treated mice and its underlying mechanism. Results indicated that PSG-1 treatment significantly alleviated DOX-induced myocardial damage via attenuating apoptosis and maintaining the structure of myocardial mitochondria. Meanwhile, PSG-1-evoked cardioprotection was associated with an increase of manganese superoxide dismutase activity and decrease of caspases activities. Moreover, administration of PSG-1 suppressed DOX-induced mitochondrial disorders, which was evidenced by reducing reactive oxygen species, elevating mitochondrial membrane potential and inhibiting the opening of mitochondrial permeability transition pore. PSG-1 was also found to reduce the release of cytochrome c from mitochondria to cytoplasm in mice subjected to DOX. Finally, our findings have provided comprehensive evidence for the cardioprotective effects of PSG-1 via reduction of apoptosis mediated by modification of the mitochondrial intrinsic apoptotic pathway, indicating that PSG-1 could be developed as an effective therapeutic strategy to prevent DOX-induced cardiotoxicity in clinical settings.

Development of a TaqMan-based real-time RT-PCR assay for the detection of SADS-CoV associated with severe diarrhea disease in pigs.

Swine acute diarrhea syndrome coronavirus (SADS-CoV) is a novel coronavirus which was first reported in southern China in 2017. It can cause severe diarrhea disease in pigs. In order to detect this new emerging virus rapidly and reliably, a TaqMan-based real-time RT-PCR assay was established in this study. Specific primers and probe were designed and synthesized based on the conserved region within the N gene of the viral genome. Results showed that the lowest limit of detection was 3.0 × 101 copies/µL. This approach was specific for SADS-CoV, and there were no cross-reaction observed against other 15 swine viruses. It was 10 times more sensitive than the conventional PCR and gave higher SADS-CoV positive detection rate (70.69%, 123/174) than the conventional PCR (51.15%, 89/174) from clinical samples. These data indicated that the TaqMan-based real-time RT-PCR assay established here was an effective method with high sensitivity, specificity and reproducibility for faster and more accurate detection and quantification of SADS-CoV.

Phylogenetic and Genome Analysis of 17 Novel Senecavirus A Isolates in Guangdong Province, 2017.

Senecavirus A (SVA), an emerging RNA virus, is considered to be associated with porcine idiopathic vesicular disease (PIVD). From February to September 2017, 17 novel SVA strains were isolated from samples with the vesicular disease from Guangdong Province, China. Full-length genomes and individual genes of the 17 new SVA isolates were genetically and phylogentically analyzed. Results showed that complete genomes, VP1, 3C, and 3D genes of these 17 novel SVA isolates revealed 96.5-99.8%, 95.1-99.9%, 95.6-100%, and 96.9-99.7% nucleotides identities, respectively. Phylogenetic analyses based on sequences of full-length genomes, VP1, 3C, and 3D genes indicated that 17 novel SVA isolates separated to three well-defined groups. Meanwhile, phylogenetic analysis for all available Chinese SVA strains demonstrated that 45 Chinese SVA strains clustered into five distinct groups with no significant relationship between strains from different provinces and/or years, including a newly emerging branch in China. This is the first comprehensive study of phylogenetic analysis for all available Chinese SVA strains, indicating the appearance of a new type of SVA strains and the complicated circulations with at least five different types of SVA strains in pigs in China.

Multilevel selection of bcrABDR-mediated bacitracin resistance in Enterococcus faecalis from chicken farms.

In this study we isolated 109 Enterococcus faecalis from chicken faecal samples in 6 provinces of China to investigate the prevalence and transmission mechanism of the bacitracin resistance locus bcrABDR in E. faecalis. Thirty-seven bcrABDR-positive E. faecalis were detected with 26 different PFGE clusters. The MLST of 14 positive strains belonged to ST16 and we also detected three new sequence types. S1-PFGE analysis indicated that the locus was located on plasmids presenting different sizes, with the most prevalent size being ~50 kb (13/37). Sequence analysis revealed that 17 out of the 37 strains harbored a 5400-bp central region, in which locus bcrABDR was bracketed by two ISEnfa1 of the same orientation. Two types of bcrABDR alleles, differing in around 10% of their sequence were found. In silico analysis showed that bcrABDR is present in a variety of bacteria including the chicken commensal Enterococcus cecorum. Our results indicate that the use of bacitracin at farms might trigger the emergence and spread of the bacitracin resistance determinant bcrABDR among human bacterial pathogens. The finding of bcrABDR in the chicken commensal E. cecorum indicates that farm animals microbiota can be an important reservoir of resistance genes with relevance for human health.