Treatment and prevention of pigeon diarrhea through the application of Lactobacillus SNK-6.

In the pigeon industry, treating and preventing diarrhea is vital because it is a serious health problem for pigeons. This study investigated the incidence of diarrhea in 3 pigeon farms in Shanghai, and analyzed the microflora through 16S rDNA high-throughput sequencing. Four strains of Escherichia coli (E. coli) isolated from pigeon diarrhea feces were administered via gavage to healthy pigeons, with each pigeon receiving 2 × 108 CFU. Pigeons that developed diarrhea after E. coli challenge were treated with 3 g of Lactobacillus salivarius SNK-6 (L. salivarius SNK-6) health sand (1.6 × 107 CFU/g). Then, a mass feeding experiment expanded to 688 pairs of pigeons with 3 replicates, each receiving 3 g of health sand containing L. salivarius SNK-6 (1.6 × 107 CFU/g) every 2 wk, and fecal status monitored and recorded. The study found that the relative abundance of the Lactobacillus genus and L. salivarius in feces from pigeons with diarrhea was significantly lower than in normal pigeon feces (P < 0.05). In contrast, E. coli showed a higher abundance and diversity in feces from pigeons with diarrhea than in normal feces (P < 0.05). Three out of the 4 isolated E. coli strains caused pigeon diarrhea, resulting in a significant reduction in microbial diversity in fecal samples (P < 0.05). Both the small group attack experiment and the mass-fed additive experiment in pigeon farms demonstrated that feeding L. salivarius SNK-6 effectively cured and prevented diarrhea. Pigeons fed with L. salivarius SNK-6 exhibited no diarrhea, while the control group had a 10% diarrhea rate. In summary, a deficiency of Lactobacillus or a high abundance of E. coli in the intestine could easily cause pigeon diarrhea. Feeding L. salivarius SNK-6 could treat pigeon diarrhea, and continuous supplementation could maintain stable preventive effects.

Source and risk assessment of heavy metals in mining-affected areas in Jiangxi Province, China, based on Monte Carlo simulation.

In recent years, heavy metal contamination of soils has become a major concern in China due to the potential risks involved. To assess environmental pollution and human health risks in a typical heavy metal polluted site in Jiangxi Province, a thorough evaluation of the distribution, pollution levels, and sources of heavy metals in soils of the Yangmeijiang River watershed was conducted in this study. Positive matrix factorization and Monte Carlo simulation were used to evaluate the ecological and human health risks of heavy metals. The research findings indicate that heavy metal pollution was the most severe at the depth of 20-40 cm in soils, with local heavy metal pollution resulting from mining and sewage irrigation. The high-risk area accounted for 91.11% of the total area. However, the pollution level decreased with time due to sampling effects, rainfall, and control measures. Leaf-vegetables and rice were primarily polluted by Cd and Pb. The main four sources of heavy metals in soils were traffic emission, metal smelting, agricultural activities and natural sources, mining extraction, and electroplating industries. Heavy metals with the highest ecological risk and health risk are Cd and As, respectively. The non-carcinogenic and carcinogenic risks of children were 7.0 and 1.7 times higher than those of adults, respectively. Therefore, children are more likely to be influenced by heavy metals compared to adults. The results obtained by the risk assessments may contribute to the identification of specific sources of heavy metals (e.g., traffic emissions, metal smelting, mining excavation, and electroplating industries). Additionally, the environmental impacts and biotoxicity associated with various heavy metals (e.g., Cd and As) can also be reflected. These outcomes may serve as a scientific basis for the pollution monitoring and remediation in the mining-affected areas.

Isolation and Proteomic Analysis of Extracellular Vesicles from Lactobacillus salivarius SNK-6.

The proteins carried by the extracellular vesicles of Lactobacillus salivarius SNK-6 (LsEVs) were identified to provide a foundation for further explorations of the probiotic activities of L. salivarius SNK-6. LsEVs were isolated from the culture media of L. salivarius SNK-6 and morphological analysis was conducted by scanning electron microscopy. Subsequent transmission electron microscopy and nanoparticle tracking analysis were performed to assess the morphology and particle size of the LsEVs. In addition, the protein composition of LsEVs was analyzed using silver staining and protein mass spectrometry. Finally, internalization of the identified LsEVs was confirmed using a confocal microscope, and enzyme-linked immunosorbent assay was employed to analyze the levels of inflammatory cytokines in LPS-challenged RAW264.7 cells. The results revealed that the membrane-enclosed LsEVs were spherical, with diameters ranging from 100-250 nm. The LsEVs with diameters of 111-256 nm contained the greatest amount of cargo. In total, 320 proteins (10-38 kD) were identified in the LsEVs and included anti-inflammatory molecules, such as PrtP proteinase, co-chaperones, and elongation factor Tu, as well as some proteins involved in glycolysis/gluconeogenesis, such as fructose-1,6-bisphosphate aldolase. Enrichment analysis showed these proteins to be related to the terms "metabolic pathway," "ribosome," "glycolysis/gluconeogenesis," "carbohydrate metabolism," and "amino acid metabolism." Furthermore, the LsEVs were internalized by host liver cells and can regulate inflammation. These findings confirm that LsEVs contain various functional proteins that play important roles in energy metabolism, signal transduction, and biosynthesis.

Effect of Lactobacillus salivarius SNK-6 on egg quality, intestinal morphology, and cecal microbial community of laying hens.

The objective of this study was to investigate the effect of Lactobacillus salivarius (L. salivarius) SNK-6 supple-mentation on the laying performance, egg quality, blood parameters, intestinal morphology, and cecal microbial community of laying hens. A total of 432 healthy 30-wk-age laying hens were randomly divided into 3 groups with 6 replicates under the same husbandry and dietary regimes: control (CON); 2.0 × 108 CFU/kg L. salivarius supplementation (T1); 2.0 × 109 CFU/kg L. salivarius supplementation (T2). The experiment lasted for 10 wk. The results indicated that the supplementation resulted in a significant reduction in the broken egg and unqualified egg ratios, and a significant increase in the eggshell strength, eggshell relative weight, albumen height, and Haugh units (P < 0.05). The L. salivarius-treated hens exhibited significantly reduced serum malondialdehyde levels (P < 0.05); significantly increased total protein, phosphorus, calcitonin, and immunoglobulin M (P < 0.05); significantly increased cecal secretory immunoglobulin A concentration (P < 0.05); significantly improved villus height (VH) in the duodenum and VH to crypt depth ratio in the jejunum (P < 0.05). The serum globulin and interleukin-1ß, immunoglobulin G concentrations, and catalase activity significantly increased in T2 (P < 0.05). Furthermore, the serum interferon-α level in T1 was significantly higher than that of the CON (P < 0.05). The intestinal barrier-related mRNA gene ZO-1, CLDN1, and MUC2 expression in the jejunum was significantly upregulated in the T1 and T2 groups (P < 0.05). The Firmicutes/Bacteroidetes ratio was higher and the relative abundances of Flavonifractor and Clostridiales_noname were significantly higher in the T1 group (P < 0.05). In conclusion, dietary supplementation with L. salivarius SNK-6 may improve hen egg quality, serum antioxidant capacity, immune function, and intestinal health.

Analytical model for organic contaminant transport in a cut-off wall and aquifer dual-domain system considering barrier arrangements.

This paper presents an analytical study of organic contaminants transport in a cut-off wall and aquifer dual-domain system, considering the effects of the inlet boundary conditions and cut-off structural arrangements. The comprehensive sensitivity analysis of parameters focusing on the breakthrough time, attenuation time and cumulative concentration are presented using the Monte Carlo simulation and Sobol global method. The simplified constant inlet boundary condition can lead to an excessively conservative prediction of the contaminant breakthrough compared to the 'finite mass' and 'decaying source' boundary conditions. The cut-off wall hydraulic performance can be enhanced by reducing the contaminant's head loss, shape factor, half-life and cut-off wall hydraulic conductivity while increasing the retardation factor. The contaminant's half-life can largely influence the maximum contaminant concentration, attenuation time and breakthrough time. For example, the maximum contaminant concentrations for T1/2 = 1.4 years and T1/2 = 100 years are 13 and 123 times greater than that for T1/2 = 0.1 year, respectively. The influence of the variation of shape factor on the breakthrough curve should be taken into consideration. Altering the structural arrangement of the cut-off wall to accommodate a smaller shape factor increases the contaminant breakthrough time. The proposed solution allows the analysis of a cut-off wall and aquifer system with different inlet boundary conditions and structural arrangements of the cut-off wall.

Numerical simulation of different pollutant control measures around an old landfill contaminated site: A field scale study.

The remediation of contaminated soils is a great challenge for global environmental sciences and engineering. The landfill was a kind of infrastructure to deal with waste from different sources while it would also cause the threat to groundwater. Cut-off walls and pumping wells were usually applied in the landfill to prevent the spread of pollutants to wider areas. However, the combination of using both of methods was rarely analyzed, especially using field data for calibrating and fitting groundwater flow and pollutant transport. 7 monitoring wells were arranged in the study area to survey the subsurface seepage. The pollution monitoring was carried out for a period of 50 days, covering 31 types of inorganic and organic pollutants. The concentration of 2,4,6-trichlorophenol (TCP) was 556.7 times greater than the standard concentration. A coupled numerical model of groundwater flow and pollutant transport was developed to assess the effectiveness of various control methods. Three options were tested, including the implementation of a single cut-off wall as well as a combination of a cut-off wall and a pumping well, for preventing the discharge of pollutants from landfills. The combination of a cut-off wall and a pumping well is the best strategy for removal of TCP. The combination approaches lead to a reduction of pollution plumes by a factor of 11 compared to the case without pollution control measures. The research findings may provide a basis and reference for the application of cutoff walls and pumping well in landfill sites or contaminated groundwater.

A mobile-immobile model for contaminant transport through GCL/AL composite liner: analytical solutions.

The current study focuses on developing alternative formulations (mobile-immobile model) of transient analytical modelling for organic contaminant transport in a composite of geosynthetic clay (GCL) and attenuation layer (AL) system. The Laplace transform method is adopted to derive the solution. The proposed analytical frameworks are validated by two set of benchmarks. The varied examples of organic contaminant transport in the composite liner are evaluated by the proposed solution to discuss the effects of soil properties, flow conditions, adsorption, and mass transfer in mobile and immobile zones on the overall transport of contaminants. Finally, an example case is presented to show the application prospect of the proposed model. The result demonstrated that mass transfer between mobile and immobile may increase the breakthrough time by a factor of 2. This indicates that heterogeneities of clay are a non-negligible part for the performance assessment of the liner system. Péclet number in GCL ([Formula: see text]) is used to investigate the relative importance of advection and diffusion mechanisms. Increasing [Formula: see text] from 0.1 to 1 can lead to an increase of the breakthrough time by a factor of 15. The analytical solutions presented here may also serve as the basic benchmark test tool for alternative numerical studies of contaminants transport in heterogeneous media.

Field investigation of temporal variation and diffusion of hydrogen sulfide on waste working face and intermediate landfill cover.

This paper presents the study on the variation, influencing factors and diffusion regularity of hydrogen sulfide (H2S) concentration and surface flux on the working face and intermediate geomembrane cover of a landfill. Field investigations were conducted using static chambers at a large-scale municipal solid waste landfill in Hangzhou, China, from January 2019 to June 2021. The analytical models of H2S transport in the working face and intermediate cover were developed to investigate the surface flux under various conditions. The CALPUFF model was used to demonstrate the diffusion path. The H2S surface flux on the working face ranged from 7.1 × 10-3 to 1.7 mg/m2/h, whereas the range was found to be 1.5 × 10-4 to 0.9 mg/m2/h on the intermediate geomembrane cover. This observation indicated that the geomembrane can reduce H2S emissions. In addition, the H2S surface fluxes at the HDPE GMB seams and near the gas collecting wells were generally 1-2 orders of magnitude larger than that in the intact GMB. The analytical model estimates that the intact GMB exhibits a diffusion coefficient of H2S ranging from 2.7 × 10-11 to 2.2 × 10-10 m2/s. However, the diffusion coefficient increases significantly to a range of 3.3 × 10-11-9.8 × 10-7 m2/s on the GMB seams. According to CALPUFF results, only the H2S diffusion from the working face had areas exceeding the standard concentration.

Bisphenol A adsorption and transport in loess and cationic surfactant/hydrophilic polymer modified bentonite liners.

Bisphenol A (BPA) is a toxic endocrine disruptor often found in landfill leachate. Adsorption behaviors and mechanisms of BPA onto organo-bentonites amended loess, e.g., Hexadecyltrimethylammonium chloride-bentonite (HTMAC-B) and Carboxymethylcellulose-bentonite (CMC-B) were experimentally investigated. The adsorption capacity of loess amended by HTMAC-B (LHB) and CMC-B (LCB) is 4.2 and 4 times greater than that of loess (L), respectively. It is attributed to the increase of hydrogen bonds and hydrophobic lateral interactions between the adsorbent and the adsorbate. The binary (Pb2+-BPA) systems may enhance BPA adsorption onto the samples by the formation of coordination bonds between the hydroxyl group of BPA and Pb2+ ions. A cycled column test was used for investigating the transport behavior of BPA in LHB and LCB samples. The hydraulic conductivity of loess amended by the organo-bentonite (e.g., HTMAC-B, CMC-B) is generally lower than 1 × 10-9 m/s. Especially for CMC-B amended loess, the hydraulic conductivity can be reduced to 1 × 10-12 m/s. This guarantees the hydraulic performance of the liner system. Transport behavior of BPA in cycled column test is explained by the mobile-immobile model (MIM). Modelling results showed that loess amended by organo-bentonites can increase the breakthrough time of BPA. In comparison to loess-based liner, the breakthrough time of BPA for LHB and LCB can be increased by a factor of 10.4 and 7.5, respectively. These results indicate that organo-bentonites can serve as a potentially effective amendment for improving the adsorption of loess-based liners.

Lactobacillus salivarius SNK-6 Regulates Liver Lipid Metabolism Partly via the miR-130a-5p/MBOAT2 Pathway in a NAFLD Model of Laying Hens.

Lactobacillus spp., as probiotics, have shown efficacy in alleviating nonalcoholic fatty liver disease (NAFLD). Here, we screened a new probiotic strain, Lactobacillus salivarius SNK-6 (L. salivarius SNK-6), which was isolated from the ileum of healthy Xinyang black-feather laying hens in China. We investigated the beneficial activity of L. salivarius SNK-6 in a NAFLD model in laying hens and found that L. salivarius SNK-6 inhibited liver fat deposition and decreased serum triglyceride levels and activity of aspartate transaminase and alanine transaminase. MBOAT2 (membrane-bound O-acyltransferase domain containing 2) was directly targeted by miR-130a-5p, which was downregulated in the liver of NAFLD laying hens but reversed after L. salivarius SNK-6 treatment. Downregulation of MBOAT2, L. salivarius SNK-6 supplementation in vivo, and L. salivarius SNK-6 cell culture treatment in vitro suppressed the mRNA expression of genes involved in the PPAR/SREBP pathway. In addition, 250 metabolites were identified in the supernatants of L. salivarius SNK-6 culture media, and most of them participated in metabolic pathways, including amino acid, carbohydrate, and lipid metabolism. Targeted metabolomic analysis revealed that acetate, butyrate, and propionate were the most abundant short-chain fatty acids, while cholic acid, ursodeoxycholic acid, chenodeoxycholic acid, and tauroursodeoxycholic acid were the four most-enriched bile acids among L. salivarius SNK-6 metabolites. This may have contributed to the reparative effect of L. salivarius SNK-6 in the NAFLD chicken model. Our study suggested that L. salivarius SNK-6 alleviated liver damage partly via the miR-130a-5p/MBOAT2 signaling pathway.

Lactobacillus salivarius SNK-6 Activates Intestinal Mucosal Immune System by Regulating Cecal Microbial Community Structure in Laying Hens.

The production performance and disease resistance of laying hens decrease obviously with age. This study aimed to investigate the effects of supplementary Lactobacillus salivarius (L. salivarius) SNK-6 on laying performance, the immune-related gene expression in cecal tonsil, and the cecal microbial composition of laying hens. Here, 384 Xinyang black commercial hens (55 weeks old) were randomly allocated to three groups under the same husbandry and dietary regimes: basal diet (Con), the low L. salivarius SNK-6 group (T1: 1.0 × 106 CFU/g), and the high L. salivarius SNK-6 group (T2: 1.0 × 107 CFU/g). The results showed that the feed intake and broken-egg rate in the T1 group were significantly higher than the Con group (p < 0.05). Meanwhile, expressions of intestinal mucosal immune-related genes were significantly upregulated. The 16S rRNA gene sequencing indicated that supplementary L. salivarius SNK-6 had no significant difference in α -diversity and only displayed a trend difference in the ß-diversity of cecal microbiota (p = 0.07). LEfSe and random forest were further used to identify bacteria family Enterobacteriaceae, order RF39, genera Ochrobactrum, and Eubacterium as biomarkers between the Con and T1 groups. Genera Ochrobactrum, which had high relative abundance and nodal degree in the T1 and T2 groups, showed a significant positive correlation with the expression of TLR-6, IL-10, MHC-II, and CD40 in cecal tonsils and might play a critical role in activating the host intestinal mucosal immune responses. Overall, dietary supplementary L. salivarius SNK-6 can display an immunomodulatory function, possibly by regulating cecal microbial composition. However, the changes in immune responses may be at the expenditure of corresponding production performance, which needs to be weighed up in practical application.

The miR-216/miR-217 Cluster Regulates Lipid Metabolism in Laying Hens With Fatty Liver Syndrome via PPAR/SREBP Signaling Pathway.

Fatty liver syndrome (FLS), a common metabolic disease in laying hens, caused by excessive hepatic fat deposition is a bottleneck in the poultry industry. However, no specific therapeutic methods have been developed. Evidence suggests that microRNAs (miRNAs) are essential for liver lipid metabolism and homeostasis, providing strong evidence for targeting miRNAs as a potential treatment option for liver diseases. However, the roles of miRNAs in the pathogenesis of FLS remain unclear. In present study, RNA-sequencing was performed to discern the expression patterns of miRNAs in normal and fatty livers of laying hens. In total, 12 dysregulated miRNAs (2 down-regulated and 10 up-regulated) were detected between the normal and fatty livers. Functional enrichment analysis showed the potential impacts of the dysregulated miRNAs on lipid metabolism. Notably, miR-216a/b and miR-217-5p, which belong to the miR-216/miR-217 cluster, were up-regulated in the sera and livers of FLS chickens, as well as free fatty acid (FFA)-induced LMH cells. Oil-red O staining revealed that up-regulation of the miR-216/miR-217 cluster induced lipid accumulation in FFA-induced LMH cells. Furthermore, the dual luciferase gene reporter assay and RT-qPCR analysis demonstrated that 3-hydroxyacyl-CoA dehydratase 2, F-box protein 8, and transmembrane 9 superfamily member 3 (TM9SF3) were directly targeted by miR-216a/b and miR-217-5p, respectively, and suppressed in the fatty livers of laying hens. Moreover, overexpression of the miR-216/miR-217 cluster or reduction in TM9SF3 levels led to activation of the proliferator-activated receptor/sterol regulatory-element binding protein (PPAR/SREBP) pathway. Overall, these results demonstrate that the miR-216/miR-217 cluster regulates lipid metabolism in laying hens with FLS, which should prove helpful in the development of new interventional strategies.

Numerical model for static chamber measurement of multi-component landfill gas emissions and its application.

The quantitative assessment of landfill gas emissions is essential to assess the performance of the landfill cover and gas collection system. The relative error of the measured surface emission of landfill gas may be induced by the static flux chamber technique. This study aims to quantify effects of the size of the chamber, the insertion depth, pressure differential on the relative errors by using an integrated approach of in situ tests, and numerical modeling. A field experiment study of landfill gas emission is conducted by using a static chamber at one landfill site in Xi'an, Northwest China. Additionally, a two-dimensional axisymmetric numerical model for multi-component gas transport in the soil and the static chamber is developed based on the dusty-gas model (DGM). The proposed model is validated by the field data obtained in this study and a set of experimental data in the literature. The results show that DGM model has a better capacity to predict gas transport under a wider range of permeability compared to Blanc's method. This is due to the fact that DGM model can explain the interaction among gases (e.g., CH4, CO2, O2, and N2) and the Knudsen diffusion process while these mechanisms are not included in Blanc's model. Increasing the size and the insertion depth of static chambers can reduce the relative error for the flux of CH4 and CO2. For example, increasing the height of chambers from 0.55 to 1.1 m can decrease relative errors of CH4 and CO2 flux by 17% and 18%, respectively. Moreover, we find that gas emission fluxes for the case with positive pressure differential (∆Pin-out) are greater than that of the case without considering pressure fluctuations. The Monte Carlo method was adopted to carry out the statistical analysis for quantifying the range of relative errors. The agreement of the measured field data and predicted results demonstrated that the proposed model has the capacity to quantify the emission of landfill gas from the landfill cover systems.

A semi-analytical solution to organic contaminants transport through composite liners considering a single crack in CCL.

Compacted clay liners (CCLs) are extensively used as engineering barriers for groundwater and soil pollution. The existence of cracks/fractures in CCL caused by thermally induced shrinkage is reported to importantly damage the performance of the CCL. An analytical model is developed to study the effects of the cracks/fractures on the migration of organic contaminants through a composite liner system. Laplace transformation and Laplace inversion using the Stehfest method are adopted to derive the analytical solution, which is validated by the experimental data. The existence of crack shows a significant impact on the breakthrough curve and bottom flux of organic contaminants. Increasing the crack width from 1 to 25 mm results in an enhancement of contaminant bottom concentration by a factor of 280. Increasing the adsorption factor and degradation rate of contaminants can effectively improve the performance of the composite liner with cracks. The effects of degradation of contaminants on the breakthrough curve are found to be more significant for the case with a larger retardation factor. This may be due to the fact that increasing the retardation factor can significantly slow down the transport of contaminants, which may indirectly create a longer period for the degradation of contaminants.

An analytical model for contaminant transport in landfill composite liners considering coupled effect of consolidation, diffusion, and degradation.

One-dimensional mathematical model is developed to investigate the behavior of contaminant transport in landfill composite liner system considering coupled effect of consolidation, diffusion, and degradation. The first- and second-type bottom boundary conditions are used to derive the steady-state and quasi-steady-state analytical solutions. The concentration profiles obtained by the proposed analytical solution are in good agreement with those obtained by the laboratory tests. The bottom concentration and flux of the soil liners can be greatly reduced when the degradation effect and porosity changing are considered. For the case under steady-state, the bottom flux and concentration for the case with t 1/2 =10 years can be 2.8 and 5.5 times lower than those of the case with t 1/2 =100 years, respectively. The bottom concentration and flux of the soil liners can be greatly reduced when the coefficient of volume compressibility decreases. For quasi-steady-state and with t 1/2 = 10 years, the bottom flux and concentration for the case with m v = 0.02/MPa can be 17.4 and 21 times lower than the case with m v = 0.5/MPa. This may be due to the fact that the true fluid velocity induced by consolidation is greater for the case with high coefficient of volume compressibility. The bottom flux for the case with single compacted clay liner (CCL) can be 1.5 times larger than that for the case with GMB/CCL considering diffusion and consolidation for DCM. The proposed analytical model can be used for verification of more complicated numerical models and assessment of the coupled effect of diffusion, consolidation, and degradation on contaminant transport in landfill liner systems.

Molecular characterization and identification of a novel polymorphism of 200 bp indel associated with age at first egg of the promoter region in chicken follicle-stimulating hormone receptor (FSHR) gene.

Follicle-stimulating hormone receptor (FSHR) plays an important role in animal follicular development. Polymorphisms in FSHR promoter region likely impact transcription and follicle growth and maturation. In this study, a fragment of ~1.9 kb of cFSHR promoter for Zang, Xianju, Lohmann Brown, Jining Bairi and Wenchang breeds (line) was obtained. Totally 49 variations were revealed, of which 39 are single nucleotide substitutions, one is nucleotide substitution of (TTG) to (CAC) and nine are indels. Polymorphism at -874 site (a 200 bp indel mutation) was identified, and their effects on egg production traits as well as gene expression were analyzed. At this site, allele I(+) was dominant in Lohmann Brown and Xinyang Brown (a synthetic egg-laying line) lines, but very rare in three Chinese indigenous chicken breeds, namely Jining Bairi, Wenchang, Zang and one synthetic boiler line (Luqin). In Xinyang Brown population, the polymorphism was associated with age at first egg (AFE) (P < 0.05) and its effect on egg number at 37 weeks of age (E37) and egg number at 57 weeks of age (E57) was not significantly different (P > 0.05). The cFSHR mRNA level was not significantly different between three genotypes in small white and small yellow follicles of Xinyang Brown hens, however, allele I(+) tends to increase cFSHR transcription.