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To disclose the antimicrobial susceptibility and wide adaptability of commonly occurring genotypes of Salmonella enterica serovar Typhimurium, the antimicrobial resistance and multilocus sequence t...

An Escherichia coli strain isolated from the feces of swine at a pork slaughterhouse in Henan province China was found to possess two colistin-resistance genes, mcr-1 and mcr-3, plus 16 additional resistance genes. Genes mcr-1.1 and mcr-3.1 were identified on IncHI2 and IncX1 type plasmids, respectively. Transconjugants (containing mcr-3, mcr-1&mcr-3) were obtained that were 64- and 512-fold higher than the minimum inhibitory concentration of colistin on the recipient bacteria (E. coli C600), respectively. The IncX1 plasmid containing mcr-3.1 displayed a very specific structure compared with previous mcr-3. Variable and stable regions were similar across different plasmids, multiple insertion sequences and transposases.

In recent years, foodborne diseases caused by pathogens have been increasing. Therefore, it is essential to control the growth and transmission of pathogens. Bacteriophages (phages) have the potential to play an important role in the biological prevention, control, and treatment of these foodborne diseases due to their favorable advantages. Phages not only effectively inhibit pathogenic bacteria and prolong the shelf life of food, but also possess the advantages of specificity and an absence of chemical residues. Currently, there are many cases of phage applications in agriculture, animal disease prevention and control, food safety, and the treatment of drug-resistant disease. In this review, we summarize the recent research progress on phages against foodborne pathogenic bacteria, including Escherichia coli, Salmonella, Campylobacter, Listeria monocytogenes, Shigella, Vibrio parahaemolyticus, and Staphylococcus aureus. We also discuss the main issues and their corresponding solutions in the application of phages in the food industry. In recent years, although researchers have discovered more phages with potential applications in the food industry, most researchers use these phages based on their host spectrum, and the application environment is mostly in the laboratory. Therefore, the practical application of these phages in different aspects of the food industry may be unsatisfactory and even have some negative effects. Thus, we suggest that before using these phages, it is necessary to identify their specific receptors. Using their specific receptors as the selection basis for their application and combining phages with other phages or phages with traditional antibacterial agents may further improve their safety and application efficiency. Collectively, this review provides a theoretical reference for the basic research and application of phages in the food industry.

As a natural alternative to traditional antimicrobials, phages are being recognised as highly effective control agents for Salmonella and other foodborne bacteria. Due to the high diversity of Salmonella serotypes and the emergence of phage-resistant strains, attempting to isolate more widespread, strictly lytic Salmonella phages is highly warranted. In this study, a lytic phage, LP31, was isolated from poultry faecal samples. Transmission electron microscopy revealed that the phage had a polyhedral head and a retraction-free tail, indicative of the Siphoviridae family. Adsorption rate experiments showed that LP31 required the participation of lipopolysaccharides, but not flagella, during phage adsorption. Host profile identification showed that LP31 could lyse most Salmonella Enteritidis (S. Enteritidis) (96.15%, N = 104) and Salmonella Pullorum (S. Pullorum) (96.67%, N = 60). Initial applications found that LP31 reduced the concentration of static S. Enteritidis on metal surfaces (0.951 log

Through long-term coevolution with its host, Mycobacterium tuberculosis (M. tuberculosis) uses multiple strategies to escape host defenses. The M. tuberculosis Rv0927c protein is predicted to be a short-chain dehydrogenase/reductase related to bacterial metabolism. However, the role of Rv0927c during M. tuberculosis infection remains unclear. Here, we observed that Rv0927c inhibited the expression of IL-6, TNF-α, and IL-1β, an effect dependent on NF-κB and p38 pathways. Western blot analysis of macrophages infected with recombinant Mycobacterium smegmatis strains showed that Rv0927c attenuated NF-κB activation by downregulating the phosphorylation of IκBα. Additionally, Rv0927c enhanced intracellular survival of M. smegmatis and pathological effects in mice. In conclusion, our findings demonstrate that Rv0927c functions as a regulator of inflammatory genes and enhances the survival of M. smegmatis.

Salmonella enterica serovar Enteritidis (S. Enteritidis) is a facultative intracellular pathogen deploying the type III secretion system (T3SS) encoded by Salmonella Pathogenicity Island 2 (SPI2) to transfer effector proteins into host cells to modify its functions and accomplish intracellular replication. To study the effect of SspH2 on immune response induced by S. Enteritidis, we generated a deletion mutant of the effector gene sspH2 and a plasmid mediated complementary strain in S. Enteritidis C50336. The results of LD50 showed that SspH2 has no obvious effect on the virulence of S. Enteritidis. However, deletion of sspH2 decreased the invasion and intercellular colonization of the bacteria in Caco2 BBE cells. Using bacteriological counts from tissue homogenates the result of colonization in internal organs showed that in spleen and liver tissues, at 3rd and 4th day p.i. there is a significance decreased number of C50336-ΔsspH2 compared to the C50336-WT and C50336-ΔsspH2-psspH2, respectively. The qRT-PCR analysis results of both in vivo and in vitro experiments clearly showed that the mutant strain C50336ΔsspH2 significantly promoted expression of IL-1β, INF-γ, IL-12, and iNOS cytokines compared to the groups infected with the wild type or complementary strains, while the IL-8 synthesis was decreased in the mutant strain infected group. All of these findings revealed that SspH2 promotes the colonization of S. Enteritidis in host cells, and it is an important anti-inflammatory biased effector in Salmonella.

Salmonella 4,[5],12:i:- is a monophasic variant of Salmonella Typhimurium, which is responsible for global foodborne disease outbreaks. Here, 255 S. 4,[5],12:i:- strains isolated from humans (11.0%) or food-borne animals (89.0%) between 2010 and 2018 were examined. Tests of susceptibility to 19 antimicrobial agents using the broth micro dilution method showed that 99.2% (n = 253) of the isolates were resistant to at least one compound. Antibiotic susceptibility analysis demonstrated that 91.8% of the isolates were multidrug-resistant (MDR) strains with predominant resistance to tetracycline (90.6%), followed by resistance to ampicillin (86.3%), streptomycin (63.5%), chloramphenicol (62.7%), and trimethoprim-sulfamethoxazole (55.3%). The 5 major distinct patterns of multi-resistance were identified as R-type AST, R-type ACTSxt, R-type ACSTSxt, R-type ACGSTSxt and R-type ASTSxt. Among the PMQR genes examined in this study, oqxAB and aac (6')-Ib-cr were the most prevalent resistance genes in the multi-resistant isolates. Our findings highlight the prevalence of the resistance of S. 4,[5],12:i:- in some regions of China, and several common types of multidrug resistance phenotypes, to provide valuable information for epidemiological studies, risk management, and public health strategies.

Deaths associated with tuberculosis (TB) is rising and accounted for 1.4 million deaths in 2015 many of which were due to drug-resistant bacteria. Vaccines represent an important medical intervention, but the current Bacilli Calmette-Guerin (BCG) vaccine is not ideal for the protection of teenagers and adults. Therefore, a safe and effective vaccine is urgently needed. In this study, we designed a novel vaccine using an attenuated Listeria monocytogenes strain carrying fusion antigen FbpB-ESAT-6 (rLM) and characterized its safety and protective efficacy against Mycobacterium tuberculosis (M.tb) infection in mice. Compared to the wild type strain yzuLM4 and parental strain LMΔactA/plcB (LM1-2), the virulence of rLM was significantly reduced as judged by its infectious kinetics and LD50 dose. Further characterization of intravenous immunization showed that prime-boost vaccination significantly increased the levels of Th1 cytokines (IFN-γ, IL-17, and IL-6), and enhanced cytotoxic T lymphocyte (CTL) CTLs activity, suggesting that rLM could elicit potent Th1/Th17 responses. More importantly, rLM significantly conferred the protection against M.tb H37Rv challenge. Collectively, our findings indicated that rLM is a novel and useful tool to prevent M.tb infection, and can be potentially be used to boost BCG-primed immunity.

The tuberculosis (TB) Bacillus Calmette-Guerin (BCG) vaccine, which is based on Mycobacterium bovis, has variable protective efficacy in humans; therefore, safe and effective vaccines are urgently required to prevent this disease. The antigen 85A from Mycobacterium tuberculosis (M. tuberculosis) is a prime target for immunity in the early phase of tuberculosis in various animal models. The purpose of this study was to confirm whether Ag85A could elicit protective immune responses in mice. Dimethyldioctadecylammonium (DDA), an adjuvant with Th1-promoting activities and monophosphoryl lipid A (MPL), an immunostimulatory component that has strong adjuvant activity for both cellular and humoral immune responses, has been used as the adjuvant to study the biological and immunological characteristics of tuberculosis proteins in the researches previously. The gene coding Ag85A (fbpA), was cloned into a pET-30a(+) prokaryotic expression vector, and the induced amino acid sequence corresponds to a 33 kDa protein, which was confirmed by mass spectrometry and western blots. Mice were subcutaneously immunized with Ag85A protein emulsified with DDA and MPL and the results showed that the 85A antigen, when combined with these adjuvants, elicited strong Ag85A-specific T-cell responses and humoral responses as compared with vaccination with BCG. Based on the fact that this vaccine combination induced strong antigen-specific immune responses, it is a prime candidate as a component of a future TB vaccine. Key words: Tuberculosis, Ag85A, prokaryotic expression, cellular immune response, humoral response.

Emerging Microbes & Infections (2017) 6, e47; doi:10.1038/emi.2017.33; published online 7 June 2017