Performance and mechanism of biochar@FeMg-LDH for efficient activation of persulfate for degradation of 2, 4-dichlorophenol in groundwater.

Groundwater environments are complex, and traditional advanced oxidation technologies mainly based on free radicals have limitations such as poor selectivity and low interference resistance, making it difficult to efficiently degrade target pollutants in groundwater. Therefore, we developed a sludge-based biochar-supported FeMg-layered double hydroxide catalyst (BC@FeMg-LDH) for the catalytic degradation of 2, 4-dichlorophenol (2, 4-DCP) using persulfate (PDS) as an oxidant. The removal efficiency of the catalyst exceeded 95%, showing high oxidation activity in a wide pH range while being almost unaffected by reducing substances and ions in the environment. Meanwhile, under neutral conditions, the leaching of metal ions from BC@FeMg-LDH was minimal, thereby eliminating the risk of secondary pollution. According to quenching experiments and electron paramagnetic resonance spectroscopy, the main active species during BC@FeMg-LDH/PDS degradation of 2, 4-DCP is 1O2, indicating a non-radical reaction mechanism dominated by 1O2. Characterization techniques, including X-ray photoelectron spectroscopy and Fourier transform infrared spectroscopy, revealed that the carbonyl (C = O) and metal hydroxyl (M-OH) groups on the material surface were the main reactive sites mediating 1O2 generation. The 1O2 generation mechanism during the reaction involved ketone-like activation of carbonyl groups on the biochar surface and complexation of hydroxyl groups on the material surface with PDS, resulting in the formation of O2·- and further generation of 1O2. 1O2 exhibited high selectivity toward electron-rich organic compounds such as 2, 4-DCP and demonstrated strong interference resistance in complex groundwater environments. Therefore, BC@FeMg-LDH holds promising applications for the remediation of organic-contaminated groundwater.

Efficient adsorption of Cu2+ and Cd2+ from groundwater by MgO-modified sludge biochar in single and binary systems.

In this study, MgO-modified sludge biochar (1MBC) prepared from sewage sludge was successfully used as an efficient adsorbent to remove heavy metals from groundwater. The adsorption performance and mechanism of 1MBC on Cu2+ and Cd2+ were investigated in single and binary systems, and the contribution of different mechanisms was quantified. Adsorption kinetics and isotherms analysis revealed that the adsorption processes of Cu2+ and Cd2+ by 1MBC followed the pseudo-second-order kinetic and Langmuir isotherm model in both systems, indicating that Cu2+ and Cd2+ were mainly controlled by chemisorption, and their theoretical maximum adsorption capacities were 240.36 and 219.06 mg·g-1, respectively. The results of the binary system showed that due to the competitive adsorption, the adsorption capacity of 1MBC for both heavy metals was lower than that of the single system, and the selective adsorption of Cu2+ was higher. The influencing variable experiments revealed that the adsorption of Cu2+ and Cd2+ by 1MBC had a wide pH adaption range and strong anti-interference ability to coexisting organics and ions. The adsorption mechanisms involved ion exchange (Cu: 47.39%, Cd: 53.17%), mineral precipitation (Cu: 35.31%, Cd: 24.18%), functional group complexation (Cu: 10.44%, Cd: 14.53%), and other possible mechanisms (Cu: 6.87%, Cd: 8.12%). Furthermore, 1MBC demonstrated excellent regeneration potential after five cycle times. Overall, the results have significant reference value for the practical application of removing heavy metals.

Heterogeneity and transmission of food safety-related enterotoxigenic Staphylococcus aureus in pig abattoirs in Hubei, China.

The dissemination of Staphylococcus aureus in the pork production chain is a major food safety concern. Abattoirs can serve both as disruptor and transmitter for S. aureus. In this study, we conducted a systematic genomic epidemiology research on the prevalence, heterogeneity, and transmission of S. aureus in 3,638 samples collected from four pig abattoirs in Hubei province, China. Our findings revealed substantial heterogeneity between S. aureus recovered from samples collected at upstream (from stunning step to head-removal step) and downstream (from splitting step to chilling step) locations within the slaughter process. Overall, 966 (26.6%) samples were positive for S. aureus, with significantly higher overall prevalence for upstream samples (29.0%, 488/1,681) compared to downstream samples (24.4%, 478/1,957). Antimicrobial susceptibility testing demonstrated that the isolates from the upstream exhibited significantly higher resistance proportions to different antimicrobials than those from the downstream. Whole-genome sequencing of 126 isolates revealed that ST398 (32.9%, 23/70) and ST9 (22.9%, 16/70) were more common among upstream isolates, while ST7 (35.7%, 20/56) and ST97 (28.6%, 16/56) were most frequently observed among downstream isolates. Additionally, molecular characterization analysis demonstrated that upstream isolates possessed significantly higher enterotoxigenic potential, more antimicrobial resistance genes, and S. aureus pathogenicity islands than downstream isolates. Notably, we discovered that enterotoxigenic S. aureus could be transmitted across different slaughter stages, with knives, water, and air serving as vectors. Although slaughtering processes had a substantial effect on reducing the food safety risk posed by enterotoxigenic S. aureus, the possibility of its widespread transmission should not be disregarded. IMPORTANCE Staphylococcus aureus (S. aureus) is one of the most important foodborne pathogens, and can cause foodborne poisoning by producing enterotoxins. Pork is a preferable reservoir and its contamination often occurs during the slaughter process. Our findings revealed significant differences in the prevalence, antimicrobial resistance, and enterotoxigenic potential between the upstream and downstream isolates within the slaughter process. Also, it is imperative not to overlook enterotoxigenic S. aureus transmitted across all stages of the slaughter process, with notable vectors being knives, water, and air. These findings hold significant implications for policy-makers to reassess their surveillance projects, and underscore the importance of implementing effective control measures to minimize the risk of S. aureus contamination in pork production. Moreover, we provide a more compelling method of characterizing pathogen transmission based on core-SNPs of bacterial genomes.

[Sludge Biochar Modified by B-doped and Its Adsorption Behavior and Mechanism of 1,2-DCA in Water].

Sludge biochar (BC600) and B-doped sludge biochar (BBC600) were prepared with the boric acid doping modified co-pyrolysis method using municipal sludge as precursors, and the materials were structurally characterized by SEM, BET, FTIR, and Zeta potential and static contact angle to investigate the adsorption behavior, mechanism of BC600 and BBC600 on 1,2-DCA in water, and the influencing factors. The results of structural characterization showed that the B element content, specific surface area, and pore volume of biochar increased by 76%, 48%, and 30%, respectively, after the B doping modification; the effect of B doping modification on the surface charge and hydrophobicity of biochar was not significant. The results of adsorption experiments showed that the adsorption of 1,2-DCA by BBC600 was better than that by BC600 due to the larger specific surface area and higher strength of oxygen-containing functional groups of BBC600; the pseudo-first-order kinetic equation could better describe the adsorption of 1,2-DCA by BC600, and the pseudo-second-order kinetic equation could better fit the adsorption of 1,2-DCA by BBC600. The intraparticle diffusion was not the only rate-limiting step affecting the adsorption rate; the biochar material was more dispersed and stable under alkaline conditions, and its oxygen-containing functional groups were deprotonated and had enhanced electron-donating ability, which was beneficial to the adsorption of 1,2-DCA. Humic acid (HA) showed a low concentration-promoting and high concentration-inhibiting effect on the adsorption of 1,2-DCA by BC600, whereas both low and high concentrations of HA showed an inhibitory effect on the adsorption of 1,2-DCA by BBC600. The adsorption of 1,2-DCA by BC600 was inhibited by both low and high concentrations of HA, and HA competed with 1,2-DCA for adsorption; Cl-, SO42-, and NO3- all inhibited the adsorption of 1,2-DCA by biochar, and the degree of inhibition ordered from small to large was Cl-

Penicillin-binding protein 1b encoded by mrcB gene mediates the enhancement of biofilm formation by subinhibitory concentrations of cefotaxime in monophasic Salmonella Typhimurium strain SH16SP46.

Development of cefotaxime-resistance and biofilm formation increase the difficulty to prevent and control the infection and contamination of Salmonella, one of the most important foodborne and zoonotic bacterial pathogen. Our previous study observed that 1/8 minimum inhibitory concentration (MIC) of cefotaxime induced the enhancement of biofilm formation and filamentous morphological change by a monophasic Salmonella Typhimurium strain SH16SP46. This study was designed to explore the role of three penicillin-binding proteins (PBPs) in mediating the induction effect of cefotaxime. Three deletion mutants of the genes mrcA, mrcB, and ftsI, encoding the proteins PBP1a, PBP1b, and PBP3, respectively, were constructed using the parental Salmonella strain SH16SP46. Gram staining and scanning electron microscopy showed that these mutants showed normal morphology comparable to the parental strain without cefotaxime treatment. However, under the stress of 1/8 MIC of cefotaxime, the strains WT, ΔmrcA, and ΔftsI, rather than ΔmrcB, exhibited filamentous morphological change. Moreover, cefotaxime treatment significantly enhanced biofilm formation by the strains WT, ΔmrcA, and ΔftsI, but not by the ΔmrcB strain. The complement of mrcB gene in the ΔmrcB strain recovered the enhanced biofilm formation and filamentous morphological change induced by cefotaxime. Our results suggest that PBP1b encoded by mrcB gene may be a binding target of cefotaxime for initiating the effect on Salmonella morphology and biofilm formation. The study will contribute to further understanding of the regulatory mechanism of cefotaxime on Salmonella biofilm formation.

Cotransfer of resistance to cephalosporins, colistin, and fosfomycin mediated by an IncHI2/pSH16G4928-like plasmid in ESBL-producing monophasic Salmonella Typhimurium strains of pig origin.

AIMS: To assess the co-transmission risk of phenotypic and genetic resistance to cephalosporins, colistin, and fosfomycin in Salmonella strains collected along the whole pork production chain. METHODS AND RESULTS: From a total of 107 Salmonella isolates from samples collected in pig slaughterhouses and markets, 15 ESBL-producing Salmonella strains resistant to cefotaxime were identified by broth microdilution method and clavulanic acid inhibition test, including 14 monophasic Salmonella Typhimurium strains and one Salmonella Derby strain. Whole genome sequence analysis showed that nine monophasic S. Typhimurium strains coresistant to colistin and fosfomycin carried the resistance genes blaCTX-M-14, mcr-1, and fosA3. Conjugational transfer tests demonstrated that the phenotypic and genetic resistance to cephalosporins, colistin, and fosfomycin could cotransfer back and forth between Salmonella and Escherichia coli via an IncHI2/pSH16G4928-like plasmid. CONCLUSIONS: This study reports the cotransmission of phenotypic and genetic resistance to cephalosporins, colistin, and fosfomycin via an IncHI2/pSH16G4928-like plasmid in Salmonella strains of animal origin, giving an alarm for the prevention of the development and spread of bacterial multidrug resistance.

Prevalence and Virulence Determinants of Staphylococcus aureus in Wholesale and Retail Pork in Wuhan, Central China.

Staphylococcus aureus is one of the major foodborne pathogens and can cause serious foodborne illness in humans by foods contaminated with S. aureus enterotoxins. In recent years, livestock-associated S. aureus has been a major public health concern for humans and has emerged in various countries globally. China is one of the largest producers of pigs and pork in the world. However, there are few studies on the detailed genotypic and pathogenic characterization of pork-associated S. aureus in China. In this study, the prevalence, antimicrobial resistance, and genotypic characteristics of S. aureus in raw pork in Wuhan, China, were investigated through multilocus sequence typing (MLST), staphylococcal protein A gene (spa) typing, and whole-genome sequencing analysis. A total of 518 S. aureus isolates (16.9%) were isolated from 3067 retail and wholesale pork samples. The prevalence of S. aureus in retail pork (22.7%) was significantly higher than in wholesale pork (15.1%), while the proportion of multidrug-resistant (MDR) isolates in wholesale pork (12.9%) was significantly higher than in retail pork (6.2%). Among the isolates, 10.8% were resistant to three or more antibiotics, with higher rates of resistance to penicillin (88.8%) and erythromycin (58.1%). A total of 28 sequence types (STs) were identified in the 518 isolates, and the predominant type was ST7 (57.5%), followed by ST5 (9.1%). In addition, based on the whole-genome sequences of 39 representative strains, 17 spa types were identified among the isolates, of which t899, t091, and t437 were the most common. Furthermore, 19 staphylococcal enterotoxin (SE) and SE-like (SEl) toxin genes were detected in the isolates, of which selw was the most common type (100%), followed by sei, sem, seo, seu, and selv (46.2%); sey (35.9%); and sea, seg, and sen (33.3%). This study found for the first time that ST7-t091-selw and ST9-t899-SCCmecXII-selw were the predominant genotypes of S. aureus in pork in China, which indicated the spreading of S. aureus with multiple virulence factors, especially with new SE/SEl types in pigs and pork, is a serious new challenge for food safety. Good hygiene and good production practices to prevent interspecies transmission and cross-contamination of S. aureus in the pig-pork chain are of great significance to public health.

Sub-inhibitory concentrations of cefotaxime treatment enhances biofilm formation of monophasic Salmonella Typhimurium variant strain SH16SP46.

Salmonella is one of the most important foodborne and zoonotic pathogens, and monophasic S. Typhimurium is ranked among the top-five Salmonella serovars causing animal and human infections worldwide. Resistance to the third- and higher-generation cephalosporins in Salmonella has attracted great attention. Bacteria are frequently exposed to sub-minimal inhibitory concentrations (sub-MICs) of antimicrobials that can trigger diverse adaptive responses such as biofilm formation. Biofilms can promote bacterial defense to external and internal harsh conditions. This study aimed to investigate the effect of sub-MICs of cefotaxime, one of the third-generation cephalosporins, on biofilm formation by non-clinical S. enterica strains. Crystal violet staining demonstrated that cefotaxime at 1/8 MIC enhanced biofilm formation by two monophasic S. Typhimurium strains. Confocal laser scanning microscopy and enzymatic treatment assay revealed that cellulose was the most dominant extracellular matrix component contributing to Salmonella biofilm formation. Scanning electron microscopy demonstrated that cefotaxime treatment led to bacterial incomplete cell division and filamentous morphology during the whole process of biofilm formation. Our study is the first to report the enhancement effect of cefotaxime on non-clinical, monophasic S. Typhimurium by affecting bacterial morphology. The results will contribute to conducting risk assessments of Salmonella in the pork production chain and guiding the rational use of antimicrobial agents to reduce the risk of biofilm formation.

Risk factors for lumbar disc herniation in adolescents and young adults: A case-control study.

Background: There is a limited understanding of the risk factors for lumbar disc herniation (LDH) in younger people, even though the evidence suggests that LDH is more prevalent in this population. This study aimed to comprehensively analyze the risk factors for LDH in adolescents and young adults. Methods: The medical records of all patients were retrospectively reviewed with inclusion criteria of being younger than 25 years. Magnetic resonance imaging (MRI) was used to confirm LDH from September 2016 to September 2021. Furthermore, 104 healthy people in the same age range were enrolled as the control group from physical examination centers. Gender, BMI, smoking, drinking, genetic history, sitting posture, daily sitting time, traumatic history of the lower back, scoliosis, and daily exercise time were examined for all enrolled people. These factors were statistically analyzed to determine the high-risk factors. Results: A total of 208 young individuals were enrolled in the present study. The mean age of the study group and the control group was 21.06 ± 3.27 years (range: 11-25 years) and 21.26 ± 2.23 years (range: 15-25 years), respectively. The result of the chi-squared test demonstrated that there was a significant difference in BMI of more than 30 (p < 0.001), genetic history (p = 0.004), sitting posture (p < 0.001), daily sitting time of more than 6 h (p < 0.001), and the history of low back trauma (p = 0.002). Additionally, multivariate logistic regression showed that these were high-risk factors for LDH, particularly the duration of daily sitting time (more than 6 h). Conclusions: BMI of more than 30, genetic history, sitting posture, daily sitting time of more than 6 h, and a history of low back trauma are the high-risk factors for adolescents and young adults with LDH. Therefore, providing them with the proper guidance and education, particularly about the protection of the lower back and the reduction of spinal load, could play a key role in preventing and reducing LDH.

Prevalence and Antimicrobial Resistance of Salmonella and Staphylococcus aureus in Fattening Pigs in Hubei Province, China.

Pig is usually the carrier of Salmonella and Staphylococcus aureus, and can transmit the bacteria along the pork production chain to cause severe public health problems. In this study, we investigated the prevalence of Salmonella and S. aureus in fattening pigs in Hubei Province, China. The overall prevalence of Salmonella in rectal swab among 896 samples from 22 farms was 17.30%, and that of S. aureus in nasal swab among 814 samples from 20 farms was 28.26%. Antimicrobial resistance (AMR) analysis showed that 95.33% of the Salmonella strains exhibited resistance to more than three classes of antimicrobial agents tested. The highest resistance proportions were for chloramphenicol (100%), sulfamethoxazole/trimethoprim (SXT) (100%), and tetracycline (TET) (93.46%), while the lowest proportions were for cefotaxime (37.38%), gentamicin (GEN) (34.58%), and ciprofloxacin (24.30%). On the other hand, 98.42% of the S. aureus strains were resistant to more than three classes of antimicrobial agents tested. The most common resistance among the S. aureus strains was against SXT (100.00%), followed by TET (98.43%), erythromycin (91.34%), and clindamycin (91.34%), while the lowest frequent resistances were against GEN (34.65%) and oxacillin (16.54%). The prevalence and AMR of Salmonella and S. aureus exhibited an obvious diversity among different pig farms. Our results provided the epidemiological data for risk analysis of foodborne bacteria and AMR in pig farms.

[Mechanism and Influencing Factors of Increasing Soil Temperature by in-situ Electrical Resistance Heating].

The mechanism and influencing factors of an in-situ thermal remediation using electrical resistance heating were investigated. The effects of electrical current, heating method, rehydration, and negative pressure on soil heating and energy consumption were studied using in-situ electrical resistance heating equipment. The results showed that there were two main mechanisms for soil heating. Firstly, electric energy was converted into heat energy, whereby direct heating of the soil by electricity increased the soil temperature. This mechanism mainly existed in the soil between two electrodes. The second was heat conduction, whereby the soil temperature between the electrodes was the highest, and the heat gradually transferred from the high-temperature soil to the low-temperature soil, such that the temperature of the soil far away from the electrode connection gradually increased. The heating current affected the rate of increase of the soil temperature. The higher the current was, the faster the soil temperature rate of increase was and the lower the unit energy consumption was. Compared with continuous heating, intermittent heating had a slower heating rate and required a longer time to reach the same temperature. However, the energy consumption per unit was low and only 45.2% of that of continuous heating. During the heating process, water should be continuously added to the soil around the electrode to maintain a high current and continuous heating. The negative pressure of extraction was large, the soil heat loss was large, and the unit energy consumption was high. In actual projects, appropriate technological conditions should be selected according to time, cost, and the removal rate as a means of improving the efficiency of the in-situ resistance thermal desorption remediation of contaminated soil, reducing energy consumption, and shortening the time limit.

Prevalence and Characterization of Salmonella in Three Typical Commercial Pig Abattoirs in Wuhan, China.

This study was designed to investigate the prevalence and characteristics of Salmonella in three Chinese pig abattoirs (A, B, and C) in Wuhan city in 2016. Four types of pig samples were collected and cultured for Salmonella. Salmonella was detected from 329 samples among the 1440 tested (22.9%). There was no significant difference in the overall prevalence between the first visit and the second visit and among the three abattoirs. Rectal swabs (RS) exhibited a significantly higher prevalence than carcass swabs and pork. A total of 177 isolates were characterized by multilocus sequence typing, serotyping, and antimicrobial susceptibility testing. Among 17 sequence types (STs) and 13 serotypes detected, ST40, ST469, and ST34, corresponding to serovars Derby, Rissen, and Typhimurium, respectively, were predominant. The isolates from different abattoirs exhibited diverse ST distribution. The minimum inhibitory concentrations were determined using the microdilution broth method. Resistance to at least one of the antimicrobials was observed for 96.6% of the strains (171/177), and multidrug resistant (MDR) isolates accounted for 75.7% of the strains (134/177). The highest resistance proportion was for tetracycline (92.7%), and the lowest was for cefotaxime (14.1%). The isolates from abattoir A exhibited a significantly lower MDR proportion than those from other abattoirs (p < 0.05). The isolates recovered from RS and pork samples exhibited significantly higher MDR proportions than those recovered from carcass swab samples. Notably, among three predominant STs of isolates, the ST34 isolates showed the highest MDR proportion. In view of the high Salmonella prevalence and antimicrobial resistance, great attention must be paid to the monitoring and controlling of Salmonella in a full pork production chain.

Identification of an Integrase That Responsible for Precise Integration and Excision of Riemerella anatipestifer Genomic Island.

Riemerella anatipestifer is a Gram-negative, pathogenic bacterium, which is harmful to poultry. However, the genomic islands (GIs) in R. anatipestifer are not well-studied. In this study, a 10K genomic island was predicted by the bioinformatics analysis of R. anatipestifer ATCC 11845, which is widely found in other R. anatipestifer genomes. We had first reported the genomic island integration and excision function in R. anatipestifer. We successfully constructed the integration plasmid by using the integrase and 53 bp attP elements. The 10K GI was found integrated at the 53 bp attB located in the Arg-tRNA of the R. anatipestifer RA-YM chromosome. We identified an integrase that helped in the precise integration and excision in R. anatipestifer and elucidated the molecular mechanism of the 10K genomic island integration and excision. Furthermore, we provided a new method for the gene expression and construction of complementary strain.

Transmissible Gastroenteritis Virus Infection Up-Regulates FcRn Expression via Nucleocapsid Protein and Secretion of TGF-ß in Porcine Intestinal Epithelial Cells.

Transmissible gastroenteritis virus (TGEV) is a porcine intestinal coronavirus that causes fatal severe watery diarrhea in piglets. The neonatal Fc receptor (FcRn) is the only IgG transport receptor, its expression on mucosal surfaces is triggered upon viral stimulation, which significantly enhances mucosal immunity. We utilized TGEV as a model pathogen to explore the role of FcRn in resisting viral invasion in overall intestinal mucosal immunity. TGEV induced FcRn expression by activating NF-κB signaling in porcine small intestinal epithelial (IPEC-J2) cells, however, the underlying mechanisms are unclear. First, using small interfering RNAs, we found that TGEV up-regulated FcRn expression via TLR3, TLR9 and RIG-I. Moreover, TGEV induced IL-1ß, IL-6, IL-8, TGF-ß, and TNF-α production. TGF-ß-stimulated IPEC-J2 cells highly up-regulated FcRn expression, while treatment with a JNK-specific inhibitor down-regulated the expression. TGEV nucleocapsid (N) protein also enhanced FcRn promoter activity via the NF-κB signaling pathway and its central region (aa 128-252) was essential for FcRn activation. Additionally, N protein-mediated FcRn up-regulation promotes IgG transcytosis. Thus, TGEV N protein and TGF-ß up-regulated FcRn expression, further clarifying the molecular mechanism of up-regulation of FcRn expression by TGEV.

Co-infection with porcine bocavirus and porcine circovirus 2 affects inflammatory cytokine production and tight junctions of IPEC-J2 cells.

Porcine bocavirus (PBoV) has a high prevalence in both healthy and diseased swine around the world. It was recently reported that PBoV and porcine circovirus type 2 (PCV2)-which contribute to porcine diarrheal disease-have a high rate of co-infection. To clarify the pathogenesis of PBoV, we examined the co-infection rate and effects of these two pathogens in IPEC-J2 porcine intestinal enterocytes. Both single and co-infection had cytopathic effects in IPEC-J2 cells. The apoptosis and proliferation rates of cells infected with both viruses did not differ significantly from those of cells infected with either one alone. PBoV and PCV2 induced the upregulation of inflammatory cytokines and the downregulation of the tight junction proteins occludin and claudin 1 in the early stage of infection, leading to destruction of epithelial barrier integrity and enhanced cytotoxicity. These findings provide insight into the pathogenic mechanisms of PBoV and PCV2 and a basis for developing effective strategies to prevent the spread of gastrointestinal diseases in pigs and other livestock.

Overexpression of Outer Membrane Protein X (OmpX) Compensates for the Effect of TolC Inactivation on Biofilm Formation and Curli Production in Extraintestinal Pathogenic Escherichia coli (ExPEC).

Our previous study showed that the inactivation of the efflux pump TolC could abolish biofilm formation and curli production of extraintestinal pathogenic Escherichia coli (ExPEC) strain PPECC42 under hyper-osmotic conditions. In this study we investigated the role of OmpX in biofilm formation and curli production of ExPEC PPECC42. Our data showed that OmpX disruption or overexpression didn't significantly affect the biofilm formation and curli production of the wild-type strain. However, in the tolC-deleted mutant, overexpressing OmpX suppressed the effect of TolC inactivation on ExPEC biofilm formation and curli production under hyper-osmotic growth conditions. Real-time qRT-PCR confirmed that OmpX overexpression affected curli production by regulating the transcription of the curli biosynthesis-related genes in the ΔtolC strain. Our findings suggest that OmpX is involved in biofilm formation and curli production.

Phenotypic and Genotypic Resistance of Salmonella Isolates from Healthy and Diseased Pigs in China During 2008-2015.

The antimicrobial resistance of Salmonella strains is rapidly increasing worldwide, which poses significant threats to animal and public health. In this study, a total of 249 porcine Salmonella isolates collected in China during 2008-2015 were examined, including 155 clinical isolates from diseased pigs and 94 nonclinical isolates from healthy pigs. Based on the minimum inhibitory concentration of seven antimicrobial agents, 96.4% of the isolates were resistant to at least one of the tested antibiotics and 81.0% of them showed multidrug resistance. The highest antimicrobial resistance was observed for tetracycline (85.9%), and the lowest was found for cefotaxime (13.3%). The isolates from diseased pigs exhibited significantly higher levels of antimicrobial resistance than those from healthy pigs. Twenty-two isolates from healthy pigs were resistant to ciprofloxacin, which may inhibit the curative effectiveness of fluoroquinolones on bacterial food-borne poisoning and infections in humans caused by contaminated food. Moreover, cefotaxime resistance of the strains isolated from diseased pigs during 2013-2015 was significantly higher compared with the strains isolated during 2008-2010. Further study showed that the correlation between phenotypic and genotypic resistance varied among the isolates from different sources, and in many cases, the presence of resistance genes was not consistent with the resistance to the corresponding antimicrobials. These results are very significant for veterinary practice and public health.

TGEV infection up-regulates FcRn expression via activation of NF-κB signaling.

It has been well characterized that the neonatal Fc receptor (FcRn) transports maternal IgG to a fetus or newborn and protects IgG from degradation. We previously reported that FcRn is expressed in a model of normal porcine intestinal epithelial cells (IPEC-J2). Transmissible gastroenteritis is an acute enteric disease of swine that is caused by transmissible gastroenteritis virus (TGEV). How porcine FcRn (pFcRn) expression is regulated by pathogenic infection remains unknown. Our research shows that IPEC-J2 cells infected with TGEV had up-regulated pFcRn expression. In addition, the NF-κB signaling pathway was activated in IPEC-J2 cells by TGEV infection. Furthermore, treatment of TGEV-infected IPEC-J2 cells with the NF-κB-specific inhibitor BAY 11-7082 resulted in down-regulation of pFcRn expression. Transient transfection of pFcRn promoter luciferase report plasmids with overexpression of NF-κB p65 transcription factor enhanced the activation of the luciferase report plasmids. We identified four NF-κB transcription factor binding sites in the promoter region of this gene using luciferase reporter system, chromatin immunoprecipitation, electromobility shift assay, and supershift analysis. Together, the data provide the first evidence that TGEV infection up-regulates pFcRn expression via activation of NF-κB signaling.

Genetic characteristics of porcine epidemic diarrhea virus in Chinese mainland, revealing genetic markers of classical and variant virulent parental/attenuated strains.

Since October 2010, porcine epidemic diarrhea (PED) caused by variant porcine epidemic diarrhea virus (PEDV) has led great economic losses to the global pig industry, especially in China. To study the genetic characteristics of PEDV strains in Chinese mainland, a total of 603 clinical samples from nine provinces/districts of Chinese mainland from January 2014 to December 2015 were collected for RT-PCR detection and 1-1323bp of S gene of 91 isolates and ORF3 gene of 46 isolates were sequenced. The results showed that the variant PEDV were the dominant pathogens of viral diarrhea diseases in these areas. Six novel variant PEDV strains (FJAX1, FJAX2, HeNPDS1, HeNPDS2, HeNPY3, and HeNPY4) with two amino acids (aa) deletion at the 56-57 aa of S protein were identified. A total of 405 Chinese PEDV strains were subjected to phylogenetic and phylogeographic analysis. The results revealed that the subgroup Va in variant PEDV group were the dominant subgroup and the spread trend of variant PEDV strains seemed to be from the southeast coastal districts to other coastal districts and interior districts. The N-terminal of S gene (1-750bp), to some extent, could represent S1 or full length S gene for phylogenetic, similarity, antigen index, hydrophilicity plot, and differentiation analyses. The 404-472bp of S gene contained the three genetic markers, i.e., "TAA" insertion at 404-405bp, "ACAGGT" deletion at 430-435bp, and "ATA" deletion at 455-457bp can be used to differentiate the classical and variant virulent parental/attenuated PEDV strains and help us to learn the infectious and genetic characteristics of PEDV strains more convenient and cheaper. This study has important implication for understanding the infectious, genetic, and evolutionary aspects of PEDV strains in Chinese mainland.

Virulence characteristics of extraintestinal pathogenic Escherichia coli deletion of gene encoding the outer membrane protein X.

Outer membrane protein X (OmpX) and its homologues have been proposed to contribute to the virulence in various bacterial species. But, their role in virulence of extraintestinal pathogenic Escherichia coli (ExPEC) is yet to be determined. This study evaluates the role of OmpX in ExPEC virulence in vitro and in vivo using a clinical strain PPECC42 of porcine origin. The ompX deletion mutant exhibited increased swimming motility and decreased adhesion to, and invasion of pulmonary epithelial A549 cell, compared to the wild-type strain. A mild increase in LD50 and distinct decrease in bacterial load in such organs as heart, liver, spleen, lung and kidney were observed in mice infected with the ompX mutant. Complementation of the complete ompX gene in trans restored the virulence of mutant strain to the level of wild-type strain. Our results reveal that OmpX contributes to ExPEC virulence, but may be not an indispensable virulence determinant.