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Background Kabuki syndrome (KS) is a rare developmental disorder characterised by multiple congenital anomalies and intellectual disability. UTX (ubiquitously transcribed tetratricopeptide repeat, X chromosome), which encodes a histone demethylase, is one of the two major pathogenic risk genes for KS. Although intellectual disability is a key phenotype of KS, the role of UTX in cognitive function remains unclear. Currently, no targeted therapies are available for KS.Aims This study aimed to investigate how UTX regulates cognition, to explore the mechanisms underlying UTX dysfunction and to identify potential molecular targets for treatment.Methods We generated UTX conditional knockout mice and found that UTX deletion downregulated calmodulin transcription by disrupting H3K27me3 (trimethylated histone H3 at lysine 27) demethylation.Results UTX-knockout mice showed decreased phosphorylation of calcium / calmodulin-dependent protein kinase II, impaired long-term potentiation and deficit in remote contextual fear memory. These effects were reversed by an Food and Drug Administration-approved drug desipramine.Conclusions Our results reveal an epigenetic mechanism underlying the important role of UTX in synaptic plasticity and cognitive function, and suggest that desipramine could be a potential treatment for KS.

Despite the global abundance of Desert sand (DS), its application in engineering remains limited. This application can help alleviate challenges related to high unit costs and the scarcity of natural fine aggregate supplies. Freeze resistance experiments were conducted on C30 concrete incorporating varying proportions of DS, including 0 %, 25 %, 50 %, 75 %, and 100 %. DS was mixed with Tuff crushed stone fine aggregate (Manufactured sand, referred to as MS). The freeze durability of the composite fine aggregate concrete was comprehensively evaluated from three perspectives: surface morphology, a macroscopic analysis including mass loss and relative dynamic elastic modulus (RDEM), and microscopic examinations encompassing pore structure observation and scanning electron microscope (SEM) imaging. Utilizing the damage degree index of the dynamic elastic modulus and the Weibull distribution model, the life prediction process was conducted to elucidate the time-varying behavior of DS-MS concrete. According to the test findings, the concrete specimen DS25-MS75 demonstrates the highest freeze resistance. The mass loss rate shows a pattern of initial decrease followed by an increase with higher desert sand admixture, whereas the RDEM initially increases and then decreases. Optimal desert sand admixture levels can prevent and delay concrete degradation, thereby enhancing its freeze resistance. Excessive desert sand content leads to increased water demand during the curing process, resulting in elevated porosity and facilitating the formation of voids. Under the action of freezing and thawing cycles in water, DS25 %-MS75 % concrete has the longest expected service life with an actual life of 101 years, while DS100 %-MS0 % concrete has the shortest service life with an actual life of only 54.5 years.

Study region: This study focuses on the Ohio Region, which spans 11 states in the eastern United States. Study focus: Runoff coefficients are crucial in hydrology, indicating the relationship between rainfall and runoff. Understanding their controls and variability is essential for water resource assessment, management strategies, and land use planning. This research examines factors influencing runoff coefficients and their trends in the Ohio region using data from the North American Land Data Assimilation System phase-2 (NLDAS-2) Mosaic Land Surface Model, covering 2000–2020. The analysis considers short-term controls, such as climatic features (rainfall intensity, amount, and duration), hydrological factors (antecedent soil moisture, drainage density, and curve number), topographic factors (drainage area, land use, slope, elevation), and watershed shape. Additionally, the study investigates trends in runoff coefficients and their long-term controls, including climatic factors and land use changes. New hydrological insights for the region: The findings indicate that runoff coefficients increase with antecedent soil moisture and rainfall intensity. Higher elevations show lower runoff coefficients due to forested land use. Larger watersheds have lower runoff coefficients at low rainfall intensity but higher ones when intensity is high. Long-term trends reveal soil moisture as the primary control, with land cover changes as a secondary factor. This research deepens understanding of runoff coefficient dynamics and controls in the Ohio region. Future studies could explore the impacts of urbanization, reservoirs, evapotranspiration, and snowmelt on runoff coefficients.

Arbuscular mycorrhizal fungi (AMF) and biochar application individually can enhance plant tolerance to saline-alkali stress and promote plant growth efficiency. However, little is known about the potential synergistic effects of their combination on improving plant growth and soil quality under saline-alkali stress. This experiment adopted the potted method to explore the effects of four treatments on switchgrass growth and soil quality: biochar (BC), Rhizophagus irregularis (Ri), biochar + Ri (BR) and a control without biochar or Ri (CK). Compared to the CK treatment, the switchgrass biomass increased by 92.4 %, 148.6 %, and 177.3 % in the BC, Ri, and BR treatment groups, respectively. Similarly, the rhizosphere soil quality index increased by 29.33 %, 22.7 %, and 49.1 % in the respective treatment groups. The BR treatment significantly altered the rhizosphere soil microbial composition and diversity. Notably, compared to the other treatments, the archaeal α-diversity in the BR group showed a significant decrease. BR treatment significantly increased the relative abundance of bacteria, fungi and archaea at the genus level (e.g., Bacillus, Trichome and candidatus_methanopenens). Network analysis showed that the complexity and closeness of interactions between different microbial taxa were stronger in the BC, Ri and BR treatments than in the CK treatment, with BR being the more prominent. In summary, biochar combined with Ri has a better effect on promoting the growth of switchgrass under saline-alkali stress, improving the quality of saline-alkali soil, and increasing soil microbial diversity. This study provides a new approach for the efficient development and utilization of saline-alkali land.

In this study, Ni-GQDs nanocomposite coatings were prepared by double-pulse electrodeposition under supercritical CO2 with graphene quantum dots (GQDs) as the second phase additive. The effects of supercritical CO2 conditions and reverse pulse current density on microstructure, crystal orientation, grain size, GQDs quality, mechanical properties, and corrosion resistance of Ni-GQDs nanocomposite coatings were investigated. The results show that when the reverse pulse current density is 0.8 A /dm2, the surface of Ni-GQDs-Ⅱ nanocomposite coating is compact and flat, GQDs is uniformly dispersed in the coating, and GQDs is closely bound to Ni grains. Compared with the coating prepared at normal temperature and pressure. The grain size of the Ni-GQDs-Ⅱ nanocomposite coating is 4.58 nm, and the grain size is reduced by 75.3 %. The quality of GQDs in the coating was improved. The coating hardness is 867.22 HV, which is significantly increased by 53.7 %. The roughness is 0.236 μm, which is significantly reduced by 37.2 %. The friction coefficient and volume wear were 0.262 and 3.395 × 107 μm3, respectively, which were significantly reduced by 27.4 % and 57.9 %. After electrochemical corrosion, the self-corrosion voltage of the coating was −139 mV, and the self-corrosion current density was 3.19 × 10−7 A/cm2. The self-corrosion voltage was significantly increased by 61.2 %, and the self-corrosion current density was significantly decreased by 71.2 %. The Rct value and Ndl value of the coating are 31594.53 Ω·cm2 and 0.862, respectively. Significantly increased by 226.2 % and 67.1 %, respectively. The coating has excellent mechanical properties and corrosion resistance.

The ecosystem of ruminal microbiota profoundly affects the health and milk production of dairy cows. High-concentrate diets are widely used in dairy farms and evoke a series of metabolic disorders. Several studies have reported the effects of high-concentrate diets on the ruminal microbiome, while the effect of changes in ruminal microbial flora, induced by high-concentrate diet feeding, on the liver of dairy cows has not been studied before. In this study, 12 mid-lactating Holstein Friesian cows (weight of 455 ± 28 kg; parities of 2.5 ± 0.5; starting milk yield of 31.59 ± 3.2 kg/d; DMI of 21.7 ± 1.1 kg/d; and a DIM at the start of the experiment of 135 ± 28 d) were fitted with ruminal fistulas, as well as with portal and hepatic vein catheters. All cows were randomly divided into 2 groups; then, they fed with low-concentrate diets (LC, concentrate: forage = 40:60) and high-concentrate diets (HC, concentrate: forage = 60:40) for 18 weeks. The forage sources were corn silage and alfalfa hay. After the cows of two groups were euthanized over two consecutive days, ruminal microbiota; the concentration of LPS in the rumen content; cecum content; the levels of blood and histamine in rumen fluid, blood, and the liver; the histopathological status of the rumen and cecum; and the inflammatory response of the liver were assessed in dairy cows under conditions of subacute ruminal acidosis (SARA). These conditions were caused by high-concentrate diet feeding. All data were analyzed using the independent t-test in SPSS. The results showed that high-concentrate diet feeding increased the concentration of LPS and histamine in the rumen and plasma of veins (p < 0.05). The abundance of Bacteroidetes at the phylum level, and of both Bacteroidetes and Saccharibacteria at the genus level, was decreased, while the abundance of Firmicutes at the phylum level and Oscillibacter at the genus level was increased by high-concentrate diet feeding. The decreased pH values of ruminal contents (LC = 6.02, HC = 5.90, p < 0.05) and the increased level of LPS in the rumen (LC = 4.921 × 105, HC = 7.855 × 105 EU/mL, p < 0.05) and cecum (LC = 11.960 × 105, HC = 13.115 × 105 EU/mL, p < 0.01) induced the histopathological destruction of the rumen and cecum, combined with the increased mRNA expression of IL-1β (p < 0.05). The histamine receptor H1R and the NF-κB signaling pathway were activated in the liver samples taken from the HC group. In conclusion, the elevated concentrations of LPS and histamine in the gut may be related to changes in the ruminal microbiota. LPS and histamine induced the inflammatory response in the ruminal epithelium, cecum epithelium, and liver. However, the cause–effect mechanism needs to be proved in future research. Our study offers a novel therapeutic strategy by manipulating ruminal microbiota and metabolism to decrease LPS and histamine release and to improve the health of dairy cows.

Milk production traits are the most important quantitative economic traits in dairy cow production; improving the yield and quality of milk is an important way to ensure the production efficiency of the dairy industry. This study carried out a series of in-depth statistical genetics studies and molecular analyses on the Chinese Holstein cows in the Jiangsu Province, such as descriptive statistics and copy number variation analysis. A genetic correlation, phenotypic correlation, and descriptive statistical analysis of five milk production traits (milk yield, milk fat percentage, milk fat yield, milk protein percentage, and milk protein yield) of the dairy cows were analyzed using the SPSS and DMU software. Through quality control, 4173 cows and their genomes were used for genomic study. Then, SNPs were detected using DNA chips, and a copy number variation (CNV) analysis was carried out to locate the quantitative trait loci (QTL) of the milk production traits by Perl program software Penn CNV and hidden Markov model (HMM). The phenotypic means of the milk yield, milk fat percentage, milk fat mass, milk protein percentage, and milk protein mass at the first trimester were lower than those at the other trimesters by 8.821%, 1.031%, 0.930%, 0.003%, and 0.826%, respectively. The five milk production traits showed a significant phenotypic positive correlation (p < 0.01) and a high genetic positive correlation among the three parities. Based on the GGPBovine 100 K SNP data, QTL-detecting research on the fist-parity milk performance of dairy cows was carried out via the CNV. We identified 1731 CNVs and 236 CNVRs in the 29 autosomes of 984 Holstein dairy cows, and 19 CNVRs were significantly associated with the milk production traits (p < 0.05). These CNVRs were analyzed via a bioinformatics analysis; a total of 13 gene ontology (GO) terms and 20 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were significantly enriched (p < 0.05), and these terms and pathways are mainly related to lipid metabolism, amino acid metabolism, and cellular catabolic processes. This study provided a theoretical basis for the molecular-marker-assisted selection of dairy cows by developing descriptive statistics on the milk production traits of dairy cows and by locating the QTL and functional genes that affect the milk production traits of first-born dairy cows. The results describe the basic status of the milk production traits of the Chinese Holstein cows in Jiangsu and locate the QTL and functional genes that affect the milk production traits of the first-born cows, providing a theoretical basis for the molecular-marker-assisted selection of dairy cows.

According to recent investigations, the proportion of mastitis caused by environmental pathogens, such as K. pneumoniae, has increased. In this research, the epidemiology of pathogens in milk samples collected from four farms in the Jiangsu Province was carried out. The results show that 16 pathogens were detected in 186 positive milk samples. It was found that K. pneumoniae had the lowest sensitivity to penicillin (0%) and amoxicillin (4%) compared to its sensitivity to gentamicin (92%) and piperacillin (89%). A total of eight ESBL-producing strains were detected. Crystal violet staining showed that 46 of the 68 isolates of K. pneumoniae had strong biofilm-forming ability, which was related to the tetracycline resistance phenotype (p < 0.05). The detection rate of the ESBL-resistant gene (blaSHV) reached 100%. The results show that resistance genes blaVIM, blaOXA-10, and blaTEM were correlated with drug-resistance phenotypes to varying degrees. The present study indicates the prevalence of bovine mastitis-derived pathogens in part of Jiangsu Province and reveals the distribution of β-lactam resistance genes and the strong biofilm-forming ability of K. pneumoniae and its relationship with tetracycline resistance. This study provided theoretical support and guidance for rational drug use and disease prevention and control on farms.

Abstract Inflammatory bowel disease (IBD) has a close association with transketolase (TKT) that links glycolysis and the pentose phosphate pathway (PPP). However, how TKT functions in the intestinal epithelium remains to be elucidated. To address this question, we specifically delete TKT in intestinal epithelial cells (IECs). IEC TKT-deficient mice are growth retarded and suffer from spontaneous colitis. TKT ablation brings about striking alterations of the intestine, including extensive mucosal erosion, aberrant tight junctions, impaired barrier function, and increased inflammatory cell infiltration. Mechanistically, TKT deficiency significantly accumulates PPP metabolites and decreases glycolytic metabolites, thereby reducing ATP production, which results in excessive apoptosis and defective intestinal barrier. Therefore, our data demonstrate that TKT serves as an essential guardian of intestinal integrity and barrier function as well as a potential therapeutic target for intestinal disorders.

As a first-line barrier against bacterial infection of mammary tissues, bovine mammary epithelial cells (bMECs) are generally believed to be involved in the immune response due to exogenous stress. Due to the escalating crisis of antibiotic resistance, there is an urgent need for new strategies to combat pathogenic bacteria-infected bovine mastitis. In this study, isolated bMECs and Institute of Cancer Research (ICR) mice were used for Escherichia coli infection and caffeic acid (CA) pretreatment experiments in vitro and in vivo. The inhibitory effect of CA on bacterial growth and biofilm formation was also demonstrated with bacteria strains isolated from mastitis-infected milk. It was demonstrated that CA supplementation prohibits the growth of the predominant strains of bacteria isolated from clinical bovine mastitis milk samples. CA was found to disrupt the biofilm formation of E. coli B1 in a sub-minimum inhibitory concentration (sub-MIC) and inhibited the adherence property of E. coli on bMECs by decreasing the staining of bacteria on cell surfaces in vitro. In addition, CA was found to attenuate proinflammatory and oxidative responses in cells infected with E. coli. The pretreatment of bMECs with CA also restored altered lipid homeostasis caused by E. coli stimulation. The protective role of CA was further confirmed via the administration of CA in mice followed by representative Gram-negative bacterial infection. Collectively, these findings highlight the potential of CA to mediate Gram-negative infections and indicate that it has the potential to be developed as a novel antibacterial drug.

Spatial interpolation techniques play an important role in hydrology, as many point observations need to be interpolated to create continuous surfaces. Despite the availability of several tools and methods for interpolating data, not all of them work consistently for hydrologic applications. One of the techniques, the Laplace Equation, which is used in hydrology for creating flownets, has rarely been used for data interpolation. The objective of this study is to examine the efficiency of Laplace formulation (LF) in interpolating data used in hydrologic applications (hydrologic data) and compare it with other widely used methods such as inverse distance weighting (IDW), natural neighbor, and ordinary kriging. The performance of LF interpolation with other methods is evaluated using quantitative measures, including root mean squared error (RMSE) and coefficient of determination (R2) for accuracy, visual assessment for surface quality, and computational cost for operational efficiency and speed. Data related to surface elevation, river bathymetry, precipitation, temperature, and soil moisture are used for different areas in the United States. RMSE and R2 results show that LF is comparable to other methods for accuracy. LF is easy to use as it requires fewer input parameters compared to inverse distance weighting (IDW) and Kriging. Computationally, LF is faster than other methods in terms of speed when the datasets are not large. Overall, LF offers a robust alternative to existing methods for interpolating various hydrologic data. Further work is required to improve its computational efficiency.

Abstract Background Lipopolysaccharides (LPS) derived from gram-negative bacterial are often regarded as primary inducer of bovine mammary inflammation. This study evaluated the biological response of metformin activated AMPK signaling on LPS-induced inflammatory responses and metabolic changes in primary bovine mammary epithelial cells (pbMEC). The pbMEC were exposed to either 3 mmol/L Metf. for 12 h as Metf. group (Metf.) or 2 μg/mL LPS for 6 h as LPS group (LPS). Cells pretreated with 3 mmol/L metformin for 12 h followed by washing and 2 μg/mL LPS exposure for 6 h were served as ML group (ML). PBS was added to cells as the control group (Con.). Results Pre-incubation with Metf. inhibited LPS-induced expression of pro-inflammatory genes (TNF, IL1B, IL6, CXCL8, MYD88 and TLR4) and proteins (IL-1β, TNF-α, NLRP3, Caspase1, ASC) and was accompanied by increased activation of AMPK signaling. Compared with the LPS group, phosphorylation of p65 and IκBα in the ML group were decreased and accumulation of NF-κB in the nucleus was significantly reduced by pretreatment with metformin. Metformin protects the cells from the increase of LPS-induced binding activity of NF-κB on both TNFA and IL1B promoters. Compared with the LPS group, genes (G6PC, PCK2) and proteins (SREBP1, SCD1) related to lipogenesis and carbohydrate metabolism were downregulated while catabolic ones (PPARA, ACSL1, Glut1, HK1) were upregulated in the ML group. Furthermore, increased acetylation of H3K14 by LPS challenge was reversed by pretreatment with metformin. Conclusion Altogether, our results indicated that pretreatment with metformin dampens LPS-induced inflammatory responses mediated in part by AMPK/NF-κB/NLRP3 signaling and modification of histone H3K14 deacetylation and metabolic changes.

Body size is one of the most economically important traits of dairy cattle, as it is significantly associated with cow longevity, production, health, fertility, and environmental adaptation. The identification and application of genetic variants using a novel genetic approach, such as genome-wide association studies (GWASs), may give more insights into the genetic architecture of complex traits. The identification of genes, single nucleotide polymorphisms (SNPs), and pathways associated with the body size traits may offer a contribution to genomic selection and long-term planning for selection in dairy cows. In this study, we performed GWAS analysis to identify the genetic markers and genes associated with four body size traits (body height, body depth, chest width, and angularity) in 1000 Chinese Holstein cows. We performed SNPs genotyping in 1000 individuals, based on the GeneSeek Genomic Profiler Bovine 100 K. In total, we identified 11 significant SNPs in association with body size traits at the threshold of Bonferroni correction (5.90 × 10−7) using the fixed and random model circulating probability unification (FarmCPU) model. Several genes within 200 kb distances (upstream or downstream) of the significant SNPs were identified as candidate genes, including MYH15, KHDRBS3, AIP, DCC, SQOR, and UBAP1L. Moreover, genes within 200 kb of the identified SNPs were significantly enriched (p ≤ 0.05) in 25 Gene Ontology terms and five Kyoto Encyclopedia of Genes and Genomes pathways. We anticipate that these results provide a foundation for understanding the genetic architecture of body size traits. They will also contribute to breeding programs and genomic selection work on Chinese Holstein cattle.

Accurately estimating the genetic parameters and revealing more genetic variants underlying milk production and quality are conducive to the genetic improvement of dairy cows. In this study, we estimate the genetic parameters of five milk-related traits of cows—namely, milk yield (MY), milk fat percentage (MFP), milk fat yield (MFY), milk protein percentage (MPP), and milk protein yield (MPY)—based on a random regression test-day model. A total of 95,375 test-day records of 9,834 cows in the lower reaches of the Yangtze River were used for the estimation. In addition, genome-wide association studies (GWASs) for these traits were conducted, based on adjusted phenotypes. The heritability, as well as the standard errors, of MY, MFP, MFY, MPP, and MPY during lactation ranged from 0.22 ± 0.02 to 0.31 ± 0.04, 0.06 ± 0.02 to 0.15 ± 0.03, 0.09 ± 0.02 to 0.28 ± 0.04, 0.07 ± 0.01 to 0.16 ± 0.03, and 0.14 ± 0.02 to 0.27 ± 0.03, respectively, and the genetic correlations between different days in milk (DIM) within lactations decreased as the time interval increased. Two, six, four, six, and three single nucleotide polymorphisms (SNPs) were detected, which explained 5.44, 12.39, 8.89, 10.65, and 7.09% of the phenotypic variation in MY, MFP, MFY, MPP, and MPY, respectively. Ten Kyoto Encyclopedia of Genes and Genomes pathways and 25 Gene Ontology terms were enriched by analyzing the nearest genes and genes within 200 kb of the detected SNPs. Moreover, 17 genes in the enrichment results that may play roles in milk production and quality were selected as candidates, including CAMK2G, WNT3A, WNT9A, PLCB4, SMAD9, PLA2G4A, ARF1, OPLAH, MGST1, CLIP1, DGAT1, PRMT6, VPS28, HSF1, MAF1, TMEM98, and F7. We hope that this study will provide useful information for in-depth understanding of the genetic architecture of milk production and quality traits, as well as contribute to the genomic selection work of dairy cows in the lower reaches of the Yangtze River.

The quality and safety of milk is challenged by cow mastitis, and the value of somatic cell score (SCS) in milk is closely related to the occurrence of mastitis. This study aimed to analyze the genetic characteristics of SCS across the first three parities in Chinese Holstein cattle, as well as to investigate potential candidate genes and biological processes that may play a potential role in the progress of cow mastitis. In this respect, we evaluated genetic parameters and conducted a genome-wide association study based on the test-day records of SCS for Chinese Holstein cows; we also validated key candidate genes using a quantitative reverse transcription PCR (RT-qPCR) experiment in primary bovine mammary epithelial cells (bMECs). The heritability of the SCS 305-day performance in milk varied between 0.07 and 0.24, and decreased with increasing parity. As the time interval grew larger, the genetic and permanent environmental correlations with the number of days in milk (DIM) weakened. Six significant single-nucleotide polymorphisms (SNPs) were identified in the association analysis, one of which was located within the exonic region of CD44. This exon-associated SNP may modify the activity of the protein encoded by the CD44. A total of 32 genes within the two hundred kilobase (kb) range of significant SNPs were detected, and these genes were markedly enriched in eight Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways and 22 biological processes, mainly participating in the progress of transmembrane transport, inflammatory factor regulation, cellular responses, the Toll-like receptor signaling pathway, and the MAPK signaling pathway. Nine genes, including the PKD2, KCNAB1, SLC35A4, SPP1, IBSP, CD14, CD44, MAPK10, and ABCG2 genes, were selected as candidate genes that could have critical functions in cow mastitis. These findings can serve as a foundation for molecular breeding and as valuable data for reducing the incidence of mastitis of Chinese Holstein cattle at the molecular level.

Bovine mastitis is often taken as one of the most common diseases in dairy farms, which its pathophysiology leads to a reduction of milk production and its quality. The penetration of pathogenic bacteria into the mammary gland, through either a contagious or environmental approach, has been determined the way of infection. The mastitis derived bacteria have become a challenge in practice, since the increasing exposure of antimicrobial. In order to identify characteristics of the epidemiological regulation and drug resistance of the pathogenic bacteria of bovine mastitis in northern Jiangsu, 156 clinical mastitis milk samples were collected from 3 large-scale farms for the epidemiological investigation and analysis of the drug resistance of E. coli. The bacteria were positively isolated in a total of 143 milk samples. The results showed that 78 strains of E. coli were detected, with a prevalence rate of 26.99%, followed by 67 strains of K. pneumoniae, with a prevalence of 23.19%, and 38 strains of Staphylococcus, with a prevalence of spp. 13.15%. In addition, 78 strains of E. coli isolated from bovine mastitis were tested for susceptibility to 8 kinds of antibiotics. It was shown that gentamicin and tetracycline were the most effective against E. coli, with the susceptibility rate of 83.3%, followed by streptomycin and ciprofloxacin, with 73.1% and 71.8% respectively. However, β-lactams including penicillin, cefothiophene, and amoxicillin showed serious resistance to E. coli isolates. There were 12 drug resistance genes detected by PCR, including β-lactam (blaTEM, blaCTX-M, and blaSHV), aminoglycoside (armA and armB), tetracycline (tetA, tetB, and tetC), and quinolone (qnrS, qepA, oqxA, and oqxB) related genes. Notably, all E. coli isolates carried blaTEM gene (100%). The detection rate of blaCTX-M was 53.8%, followed by the detection of blaSHV (20.5%), armA (9.0%), tetA (26.9%), tetB (2.6%), tetC (20.5%), qnrS (29.5%), oqxA (37.2%) and oqxB (1.3%). The present study provides crucial information on the distribution of bovine mastitis derived bacterial pathogens in Jiangsu province, as well as highlighting the antimicrobial resistance which might help to improve the efficiency of antibiotics treatment on bovine mastitis.

Bovine mastitis is perplexing the dairy industry since the initiation of intensive dairy farming, which has caused a reduction in the productivity of cows and an escalation in costs. The use of antibiotics causes a series of problems, especially the formation of bacterial antimicrobial resistance. However, there are limited antibiotic-free therapeutic strategies that can effectively relieve bacterial infection of bovine mammary glands. Hence, in this study, we constructed a mammary gland tissue-specific expression vector carrying the antimicrobial peptide of bovine-derived tracheal antimicrobial peptide (TAP) and evaluated it in both primary bovine mammary epithelial cells (pBMECs) and mice. The results showed that the vector driven by the β-lactoglobulin gene (BLG) promoter could efficiently direct the expression of TAP in pBMECs and the mammary gland tissue of mice. In addition, significant antibacterial effects were observed in both in vitro and in vivo experiments when introducing this vector to bovine-associated Staphylococcus aureus-treated pBMECs and mice, respectively. This study demonstrated that the mammary gland tissue-specific expression vector could be used to introduce antimicrobial peptide both in in vitro and in vivo and will provide a new therapeutic strategy in the treatment of bovine mastitis.

Udder conformation traits are one of the most economic traits in dairy cows, greatly affecting animal health, milk production, and producer profitability in the dairy industry. Genetic analysis of udder structure and scores have been developed in Holstein cattle. In our research, we conducted a genome-wide association study for five udder traits, including anterior udder attachment (AUA), central suspensory ligament (CSL), posterior udder attachment height (PUAH), posterior udder attachment width (PUAW), and udder depth (UD), in which the fixed and random model circulating probability unification (FarmCPU) model was applied for the association analysis. The heritability and the standard errors of these five udder traits ranged from 0.04 ± 0.00 to 0.49 ± 0.03. Phenotype data were measured from 1000 Holstein cows, and the GeneSeek Genomic Profiler (GGP) Bovine 100 K SNP chip was used to analyze genotypic data in Holstein cattle. For GWAS analysis, 984 individual cows and 84,407 single-nucleotide polymorphisms (SNPs) remained after quality control; a total of 18 SNPs were found at the GW significant threshold (p < 5.90 × 10−7). Many candidate genes were identified within 200kb upstream or downstream of the significant SNPs, which include MGST1, MGST2, MTUS1, PRKN, STXBP6, GRID2, E2F8, CDH11, FOXP1, SLF1, TMEM117, SBF2, GC, ADGRB3, and GCLC. Pathway analysis revealed that 58 Gene Ontology (GO) terms and 18 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were enriched with adjusted p values, and these GO terms and the KEGG pathway analysis were associated with biological information, metabolism, hormonal growth, and development processes. These results could give valuable biological information for the genetic architecture of udder conformation traits in dairy Holstein cattle.

Klebsiella pneumoniae (K. pneumoniae) induced bovine mastitis has been becoming one of the dominantly pathogenic bacteria in cases of bovine mastitis, and is threatening public health through dairy products. In order to explore the characteristics of multilocus sequence typing (MLST), virulence gene carrying, and the relationship between virulence genes and the antibiotic resistance of Klebsiella pneumoniae from dairy cattle in northern Jiangsu, 208 dairy milk samples were collected from four dairy farms in northern Jiangsu. A total of 68 isolates were obtained through bacterial isolation, purification, and 16S rDNA identification. Eleven virulence genes were detected by specific PCR. The susceptibility of the isolates to antimicrobials was analyzed using the Kirby–Bauer method. The Pearson correlation coefficient was used to analyze the correlation between the presence of virulence genes and the phenotype of drug resistance. ST 2661 was the most prevalent type of K. pneumoniae (13/68, 19.1%) among the 23 ST types identified from the 68 isolates. The virulence gene allS was not detected, but the positive detection rates of the virulence genes fimH, ureA, uge and wabG were 100.0%. Notably, the detection rates of genes rmpA and wcaG, related to the capsular polysaccharide, were 4.4% and 11.8%, respectively, which were lower than those of genes related to siderophores (kfuBC, ybtA and iucB at 50.0%, 23.5%, and 52.9%, respectively). The K. pneumoniae isolates were sensitive to ciprofloxacin, nitrofurantoin, and meropenem. However, the resistance rate to penicillin was the highest (58/68, 85.3%), along with resistance to amoxicillin (16/68, 23.5%). The results revealed the distribution of 23 ST types of K. pneumoniae from the milk from bovine-mastitis-infected dairy cows in northern Jiangsu, and the expression or absence of the virulence gene kfuBC was related to the sensitivity to antibiotics. The current study provides important information relating to the distribution and characteristics of K. pneumoniae isolated from dairy cows with clinical bovine mastitis, and is indicative of strategies for improving the treatment of K. pneumoniae-induced bovine mastitis.

The objective of this research was to explore the effect of metformin on the lipoteichoic acid (LTA)–induced mastitis model using isolated primary bovine mammary epithelial cells (PBMECs). The PBMECs were exposed to either 3 mM metformin for 12 h as a metformin group (MET) or 100 μg/mL LTA for 6 h as LTA group (LTA). Cells pretreated with 3 mM metformin for 12 h followed by washing and 100 μg/mL LTA exposure for 6 h served as the MET + LTA group. Phosphate-buffered saline was added to cells as the control group. PBMECs pretreated with different metformin doses were analyzed by a flow cytometry (annexin V–fluorescein isothiocyanate assay) to detect the cell apoptotic rate. We performed quantitative reverse transcriptase–polymerase chain reaction and Western blot analysis to evaluate the inflammatory and oxidative responses to metformin and LTA by measuring cellular cytotoxicity, mRNA expression, and protein expression. Immunofluorescence was used to evaluate nuclear localization. The results showed that the gene expression of COX2, IL-1β, and IL-6 significantly increased in the cells challenged with LTA doses compared to control cells. In inflammatory PBMECs, metformin attenuated LTA-induced expression of inflammatory genes nuclear factor κB (NF-κB) p65, tumor necrosis factor α, cyclooxygenase 2, and interleukin 1β, as well as the nuclear localization and phosphorylation of NF-κBp65 protein, but increased the transcription of nuclear factor erythroid 2–related factor 2 (Nrf2) and Nrf2-targeted antioxidative genes heme oxygenase-1 (HO-1) and Gpx1, as well as the nuclear localization of HO-1 protein. Importantly, metformin-induced activation of Nrf2 is AMP-activated protein kinase (AMPK)–dependent; as metformin-pretreated PBMECs activated AMPK signaling via the upregulation of phosphorylated AMPK levels, cell pretreatment with metformin also reversed the translocation of Nrf2 that was LTA inhibited. This convergence between AMPK and Nrf2 pathways is essential for the anti-inflammatory effect of metformin in LTA-stimulated PBMECs. Altogether, our results indicate that metformin exerts anti-inflammation and oxidative stress through regulation of AMPK/Nrf2/NF-κB signaling pathway, which highlights the role of AMPK as a potential therapeutic strategy for treatment of bovine mastitis.

This study aimed to evaluate whether (-)-epigallocatechin-3-gallate (EGCG) alleviates hepatic responses to lipopolysaccharide (LPS)-induced inflammation and oxidation. Isolated bovine hepatocytes and BALB/c mice were used for LPS challenge and EGCG pretreatment experiments in vitro and in vivo. LPS-challenged (6 μg/mL) hepatocytes exhibited increased levels of NF-κB (p65 and IκBα) and MAPK (p38, ERK, JNK) phosphorylation as well as increased binding activity of p65 to target pro-inflammatory gene promoters, and these effects were suppressed by pretreatment with 50 μM EGCG. Moreover, the reduction in Nrf2 signaling and antioxidant enzyme activities induced by LPS stimulation were reversed upon EGCG treatment. In vivo experiments demonstrated the protective role of EGCG in response to GalN/LPS-induced mortality and oxidative damage. Together, our results suggest that EGCG is hepatoprotective via inhibition of MAPK/NF-κB signaling and activation of the Nrf2 cascade. This information might help design strategies for counteracting hepatitis in ruminants and monogastric animals.

Escherichia coli (E. coli) is one of the main causative agents of mastitis in dairy cows. Lycium barbarum polysaccharide (LBP) has a variety of physiological effects as it has antioxidants, it is hypoglycemic, it has anti-aging properties, it is neuroprotective, immune boosting, and it has anti-inflammatory effects in vivo and in vitro. In this study, we examined whether LBP affects the expression of pro-inflammatory factors, and the mitogen-activated protein kinase (MAPK) signaling pathway via activation of the suppressor of cytokine signaling-3 (SOCS3) in E. coli-induced primary bovine mammary epithelial cell (pbMEC) inflammatory responses. The experiment was designed with the control group (NC), cells were treated with E. coli for 6 h as the E. coli group (E. coli), and cells were pretreated with 100 μg/mL or 300 μg/mL of LBP for 24 h, followed by the addition of E. coli for 6 h as the E. coli + low level (E + LL) or E. coli + high level (E + HL) groups. The addition of LBP did not alter the cell viability of pbMEC in a dose-dependent assay. Pretreatment with LBP significantly decreased the expression of pro-inflammatory genes (IL1B, MAPK14, COX-2, iNOS) and proteins (COX-2, IL-1β, TNF-α) in the cells challenged by E. coli as compared with the control group (p < 0.05). E. coli stimulation significantly increased the production of reactive oxygen species (ROS) and malondialdehyde (MDA) in pbMEC, and decreased the antioxidants’ capacity with regard to decreased superoxide dismutase (SOD) and total antioxidant capacity (T-AOC); however, pretreatment with LBP reversed the oxidative stress and inhibition of antioxidants in cells challenged by E. coli. Moreover, LBP reversed the upregulated expression of the components of the MAPK pathway (increased phosphorylation level of p38, JNK, and ERK), followed by E. coli stimulation. Consistently, cells exposed to E. coli strengthened the staining of p38, whereas pretreatment of LBP weakened the staining of p38 in cells challenged by E. coli. Notably, the expression of SOCS3 was increased by LBP added to the cells in a dose-dependent manner. Additionally, the level of decreased expression of proinflammatory factors (IL-1β, TNF-α, and COX-2) was higher in the E + LL group than in the E + HL group. These results indicate that LBP pretreatment is effective in the alleviation of E. coli-induced inflammatory and oxidative responses in pbMEC through activation of SOCS3 and depression of MAPK signaling. As such, this might help us to develop molecular strategies for mitigating the detrimental effects of clinical bovine mastitis.

Feet and leg conformation traits are considered one of the most important economical traits in dairy cattle and have a great impact on the profitability of milk production. Therefore, identifying the single nucleotide polymorphisms (SNPs), genes and pathways analysis associated with these traits might contribute to the genomic selection and long-term plan selection for dairy cattle. We conducted genome-wide association studies (GWASs) using the fixed and random model circulating probability unification (FarmCPU) method to identify SNPs associated with bone quality, heel depth, rear leg side view and rear leg rear view of Chinese Holstein cows. Phenotypic measurements were collected from 1000 individuals of Chinese Holstein cattle and the GeneSeek Genomic Profiler Bovine 100 K SNP chip was utilized for individual genotyping. After quality control, 984 individual cows and 84,906 SNPs remained for GWAS work; as a result, we identified 20 significant SNPs after Bonferroni correction. Several candidate genes were identified within distances of 200 kb upstream or downstream to the significant SNPs, including ADIPOR2, INPP4A, DNMT3A, ALDH1A2, PCDH7, XKR4 and CADPS. Further bioinformatics analyses showed 34 gene ontology terms and two signaling pathways were significantly enriched (p ≤ 0.05). Many terms and pathways are related to biological quality, metabolism and development processes; these identified SNPs and genes could provide useful information about the genetic architecture of feet and leg traits, thus improving the longevity and productivity of Chinese Holstein dairy cattle.

Reproduction is an important production activity for dairy cows, and their reproductive performance can directly affect the level of farmers’ income. To better understand the genomic regions and biological pathways of reproduction-related traits of dairy cows, in the present study, three body shape traits—Loin Strength (LS), Rump Angle (RA), and Pin Width (PW)—were selected as indicators of the reproductive ability of cows, and we conducted genome-wide association analyses on them. The heritability of these three traits was medium, ranging from 0.20 to 0.38. A total of 11 significant single-nucleotide polymorphisms (SNPs) were detected associated with these three traits. Bioinformatics analysis was performed on genes close to the significant SNPs (within 200 Kb) of LS, RA, and PW, and we found that these genes were totally enriched in 20 gene ontology terms and six KEGG signaling pathways. Finally, the five genes CDH12, TARP, PCDH9, DTHD1, and ARAP2 were selected as candidate genes that might affect LS. The six genes LOC781835, FSTL4, ATG4C, SH3BP4, DMP1, and DSPP were selected as candidate genes that might affect RA. The five genes USP6NL, CNTN3, LOC101907665, UPF2, and ECHDC3 were selected as candidate genes that might affect the PW of Chinese Holstein cows. Our results could provide useful biological information for the improvement of body shape traits and contribute to the genomic selection of Chinese Holstein cows.

Nitrogen (N) deposition and precipitation could profoundly influence the structure and function of forest ecosystems. However, conventional studies with understory additions of nitrogen and water largely ignored canopy-associated ecological processes and may have not accurately reflected the natural situations. Additionally, most studies only made sampling at one time point, overlooked temporal dynamics of ecosystem response to environmental changes. Here we carried out a field trial in a mixed deciduous forest of China with canopy addition of N and water for 4 years to investigate the effects of increased N deposition and precipitation on the diversity and community composition of arbuscular mycorrhizal (AM) fungi, the ubiquitous symbiotic fungi for the majority of terrestrial plants. We found that (1) in the 1st year, N addition, water addition and their interactions all exhibited significant influences on AM fungal community composition; (2) in the 2nd year, only water addition significantly reduced AM fungal alpha-diversity (richness and Shannon index); (3) in the next 2 years, both N addition and water addition showed no significant effect on AM fungal community composition or alpha-diversity, with an exception that water addition significantly changed AM fungal community composition in the 4th year; (4) the increment of N or water tended to decrease the abundance and richness of the dominant genus Glomus and favored other AM fungi. (5) soil pH was marginally positively related with AM fungal community composition dissimilarity, soil NH4+-N and N/P showed significant/marginal positive correlation with AM fungal alpha-diversity. We concluded that the effect of increased N deposition and precipitation on AM fungal community composition was time-dependent, mediated by soil factors, and possibly related to the sensitivity and resilience of forest ecosystem to environmental changes.

It has not been reported whether thiamine deficiency (TD) affects hippocampal neurogenesis or not. Here, we explored the influence of TD at early pre-pathological lesion stage on hippocampal neurogenesis and the correlation between affected hippocampal neurogenesis and cognitive dysfunction. We prepared TD mouse model by feeding a thiamine-depleted diet. Learning and memory functions of TD mice were tested with Y-maze. Hippocampal neurogenesis was studied with BrdU, PCNA, Dcx, and NeuN immunohistochemical staining. The results showed significant decline in learning ability and hippocampal neurogenesis simultaneously since 9–days of treatment when the model mice did not exhibit regular pathological lesion, the loss of cholinergic neurons, decrease of NeuN-positive hippocampal cell, and abnormal long-term potentiation of hippocampal CA1 and CA3. Re-administering thiamine reversed the weakened learning ability as well as the impaired hippocampal neurogenesis induced by TD at early pre-pathological lesion stage. The present study demonstrated that hippocampal neurogenesis was vulnerable to TD and the impaired hippocampal neurogenesis is greatly involved in cognitive dysfunction induced by TD at early pre-pathological lesion stage.

High concentrate diets are fed to early and mid-lactation stages dairy ruminants to meet the energy demands for high milk production in modern milk industry. The present study evaluated the effects of a high concentrate diet on milk fat and milk composition, especially, cis-9, trans-11 CLA content in milk and gene expression of lactating goats. Eight mid-lactating goats with rumen fistula were randomly assigned into a high concentrate diet (HCD) group and low concentrate diet (LCD) group. High concentrate diet feeding significantly increased lipopolysaccharides (LPS) in plasma and decreased milk fat content, vaccenic acid (VA) and cis-9, trans-11 CLA in milk of the lactating goats. The mRNA expression levels of sterol regulatory element binding protein B 1c (SREBP1c), lipoprotein lipase (LPL), fatty acid synthetase (FASN) and acetyl-CoA carboxylase α (ACACA, ACCα) involving in lipid metabolism were analyzed, and ACACA and LPL all decreased in their expression level in the mammary glands of goats fed a high concentrate diet. DNA methylation rate of stearoyl-CoA desaturase (SCD) was elevated and decreased, and SCD mRNA and protein expression was reduced significantly in the mammary glands of goats fed a high concentrate diet. In conclusion, feeding a high concentrate diet to lactating goats decreases milk fat and reduced expression of SCD in the mammary gland, which finally induced cis-9, trans-11 CLA content in milk.

PURPOSE:Epigenetic modifications critically regulate the expression of immune-related genes in response to inflammatory stimuli. It has been extensively reported that a high concentrate (HC) diet can trigger systemic inflammation in dairy cows, yet it is unclear whether epigenetic regulation is involved in the expression of immune genes in the livers of dairy cows. This study aimed to investigate the impact of epigenetic modifications on the expression of immune-related genes. EXPERIMENTAL DESIGN:In eight mid-lactating cows, we installed a rumen cannula and catheters of the portal and hepatic veins. Cows were randomly assigned to either the treatment group fed a high concentrate (HC) diet (60% concentrate + 40% forage, n = 4) or a control group fed a low concentrate (LC) diet (40% concentrate + 60% forage, n = 4). RESULTS:After 10 weeks of feeding, the rumen pH was reduced, and levels of lipopolysaccharide (LPS) in the rumen, and portal and hepatic veins were notably increased in the HC group compared with the LC group. The expression levels of detected immune response-related genes, including Toll-like receptor 4 (TLR4), cytokines, chemokines, and acute phase proteins, were significantly up-regulated in the livers of cows fed a HC diet. Chromatin loosening at the promoter region of four candidate immune-related genes (TLR4, LPS-binding protein, haptoglobin, and serum amyloid A3) was elicited, and was strongly correlated with enhanced expression of these genes in the HC group. Demethylation at the promoter region of all four candidate immune-related genes was accompanied by chromatin decompaction. CONCLUSION:After HC diet feeding, LPS derived from the digestive tract translocated to the liver via the portal vein, enhancing hepatic immune gene expression. The up-regulation of these immune genes was mediated by epigenetic mechanisms, which involve chromatin remodeling and DNA methylation. Our findings suggest that modulating epigenetic mechanisms could provide novel ways to treat systemic inflammatory responses elicited by the feeding of a HC diet.

BACKGROUND: There is current interest in understanding the molecular mechanisms of tumor-induced bone pain. Accumulated evidence shows that endogenous formaldehyde concentrations are elevated in the blood or urine of patients with breast, prostate or bladder cancer. These cancers are frequently associated with cancer pain especially after bone metastasis. It is well known that transient receptor potential vanilloid receptor 1 (TRPV1) participates in cancer pain. The present study aims to demonstrate that the tumor tissue-derived endogenous formaldehyde induces bone cancer pain via TRPV1 activation under tumor acidic environment. METHODOLOGY/PRINCIPAL FINDINGS: Endogenous formaldehyde concentration increased significantly in the cultured breast cancer cell lines in vitro, in the bone marrow of breast MRMT-1 bone cancer pain model in rats and in tissues from breast cancer and lung cancer patients in vivo. Low concentrations (1 approximately 5 mM) of formaldehyde induced pain responses in rat via TRPV1 and this pain response could be significantly enhanced by pH 6.0 (mimicking the acidic tumor microenvironment). Formaldehyde at low concentrations (1 mM to 100 mM) induced a concentration-dependent increase of [Ca(2+)]i in the freshly isolated rat dorsal root ganglion neurons and TRPV1-transfected CHO cells. Furthermore, electrophysiological experiments showed that low concentration formaldehyde-elicited TRPV1 currents could be significantly potentiated by low pH (6.0). TRPV1 antagonists and formaldehyde scavengers attenuated bone cancer pain responses. CONCLUSIONS/SIGNIFICANCE: Our data suggest that cancer tissues directly secrete endogenous formaldehyde, and this formaldehyde at low concentration induces metastatic bone cancer pain through TRPV1 activation especially under tumor acidic environment.

In this paper, a compound equation which is a mix of the Wadati--Konno--Ichikawa (WKI) equation and the short-pulse (SP) equation is first studied. By transforming both the independent and dependent variables in the equation, we introduce a novel hodograph transformation to convert the compoundWKI--SP equation into the mKdV--SG (modified Korteweg--de Vries and sine- Gordon) equation. The multiloop soliton solutions in the form of the parametric representation are found. It is shown that the N-loop soliton solution may be decomposed exactly into N separate soliton elements by using a Moloney--Hodnett-type decomposition. By virtue of the decomposed soliton solutions, the asymptotic behaviors of N = 2 and N = 3 are investigated in detail. The corresponding phase shifts of each loop or antiloop soliton caused by its interaction with the other ones are calculated. Furthermore, a new hierarchy of WKI--SP-type equations possessing multiloop soliton solutions is constructed. These deduced equations are all with time-varying coefficients and the corresponding dispersion relation will have a time-dependent velocity. The whole hierarchy of equations which include the WKI-type equations, the SP-type equations, and the compound generalized WKI--SP equations, are illustrated Lax integrable. The specific equation in the hierarchy is labeled as WKI--SP(n,m) equation so that its Lax pairs can be directly written out with the help of n and m. A unified hodograph transformation is established to relate the compound WKI--SP hierarchy with the mKdV--SG hierarchy. [ABSTRACT FROM AUTHOR]