Isolation,Identification and Analysis of Serotype and Antibiotic Susceptibility of Pathogenic Salmonella from Chickens

To investigate the situations of predominant strain and antibiotic resistance of pathogenic Salmonella from chickens in Guangxi Zhuang Autonomous region, Salmonella was pre-enriched and isolated from tissues of clinical suspected chickens, and the Salmonella isolates were identified by biochemical test using ID 32E System for the identification of Enterobacteriaceae of VITEK System ATB Expression, serotypes were determined by slid agglutination test, antibiotic susceptibility was analyzed by ATB VET Susceptibility Test Strip of Enterobacteriaceae antibiotics.The results showed that 34 Salmonella strains were obtained from 310 clinical samples, of which 1 strain belonged to A serogroup, 1 to C2 serogroup, 15 to B serogroup, 14 to D serogroup and 3 to untyped serogroup, and S.typhimurium of B serogroup and S.pullorum of D serogroup were the predominant serotypes.All Salmonella isolates were sensitive to meropenem, imipenem, spectinomycine and apramycine, and the resistance rates to chloramphenicol, kanamycine and gentamicine were less than 10%, and the resistance rates to amoxicillin, streptomycine, flumequine, oxolinique, sulfamethizol, tetracycline and nitrofurantoine ranged between 50% and 90%, while the resistance rates to penicilline, oxacilline, fusidique, rifampicine and metronidazole reached to 100%.The results indicated that the serotype distribution of pathogenic Salmonella from chickens exhibited regional characteristics, and S.typhimurium and S.pullorum were the predominant serotypes in Guangxi Zhuang Autonomous region, and antibiotic resistance of Salmonella isolates was very serious which should be highly concerned.     

Comparative Phenotypic and Genotypic Analyses of Salmonella Rissen that Originated from Food Animals in Thailand and United States

Salmonella enterica serovar Rissen has been recognized as one of the most common serovar among humans and pork production systems in different parts of the world, especially Asia. In the United States, this serovar caused outbreaks but its epidemiologic significance remains unknown. The objectives of this study were to compare the phenotypic (antimicrobial susceptibility) and genotypic attributes of Salmonella Rissen isolated in Thailand (Thai) and the United States (US). All the Thai isolates (n = 30) were recovered from swine faecal samples. The US isolates (n = 35) were recovered from swine faecal samples (n = 29), cattle (n = 2), chicken (n = 2), dog (n = 1) and a ready‐to‐eat product (n = 1). The antimicrobial susceptibility of isolates was determined using the Kirby‐Bauer disk diffusion method with a panel of 12 antimicrobials. Pulse‐field gel electrophoresis (PFGE) was used to determine the genotypic diversity of isolates. All Thai isolates showed multidrug resistance (MDR) with the most frequent antibiotic resistance shown against ampicillin (100%), sulfisoxazole (96.7%), tetracycline (93.3%), streptomycin (90%) and chloramphenicol (30%). About half of the isolates of USA origin were pan‐susceptible and roughly 30% were resistant to only tetracycline (R‐type: Te). Salmonella Rissen isolated from Thailand and the USA in this study were found to be clonally unrelated. Genotypic analyses indicated that isolates were clustered primarily based on the geographic origin implying the limited clonality among the strains. Clonal relatedness among different host species within the same geography (USA) was found. We found genotypic similarity in Thai and US isolates in few instances but with no epidemiological link. Further studies to assess propensity for increased inter‐regional transmission and dissemination is warranted.     

LysR型转录因子STM0859对鼠伤寒沙门菌环境耐受性的调控作用

为了解LysR型转录因子STM0859对鼠伤寒沙门菌(ST)环境应激的调控作用,以ST-SL1344强毒株为研究对象,利用温度敏感型质粒和λ-Red同源重组技术构建ST-ΔSTM0859基因缺失突变株,检测分析缺失株对pH、高盐、H_2O_2和乙醇胁迫环境的适应能力及在不同pH半固体中的运动能力,探究STM0859基因缺失对ST环境应激和运动能力的影响。结果显示,与ST-SL1344强毒株相比,ST-ΔSTM0859缺失株对pH=10、4%NaCl、0.1%H_2O_2和3%乙醇胁迫环境的适应能力没有显著变化,而在pH=4酸应激环境和半固体运动中生长能力极显著低于亲本株,pH=5时差异显著。提示LysR型转录因子STM0859参与ST对酸胁迫环境适应性的调控,为进一步揭示STM0859转录因子对ST环境应激调控的分子机制奠定基础。     

Proteomic analysis of intestinal mucosa responses to Salmonella enterica serovar typhimurium in naturally infected pig

Salmonella enterica serovar typhimurium (S. typhimurium) is one of the most frequent Salmonella serotypes isolated from European pigs. Despite the advances in understanding the mechanisms involved in host–pathogen interactions and host cell responses to S. typhimurium, the global change that occurs in naturally exposed populations has been poorly characterized. Here, we present a proteomics study on intestinal mucosa of pigs naturally infected with S. typhimurium, in order to better understand the pathogenesis of salmonellosis and the pathways which might be affected after infection. Samples were analyzed by 2D-DIGE and 44 different proteins exhibited statistically significant differences. The data set was analyzed by employing the Ingenuity Pathway Analysis and the physiological function most significantly perturbed were immunological and infectious disease, cellular assembly and organization and metabolism. The pathways implicated in the porcine immune response to S. typhimurium were gluconeogenesis and Rho GDI/RhoA signaling, and our results suggest that keratins and the intermediate filaments could play an important role in the damage of the mucosa and in the success of infection. The role of these findings in salmonellosis has been discussed, as well as the importance of analyzing naturally infected animals to have a complete picture of the infection. Also, we compared the results found in this work with those obtained in a similar study using experimentally infected animals.     

Attenuated Salmonella carrying pGRIM-19-si-survivin co-expression plasmid inhibits growth of prostate cancer subcutaneous xenografts in vivo

AIM：To explore the effects of pGRIM-19-si-survivin co-expression plasmid carried by human attenuated Salmonella on prostate cancer subcutaneous xenograft growth in nude mice. METHODS：Prostate cancer xenograft model was established in nude mice. Co-expression plasmids carried by attenuated Salmonella were introduced by intraperitoneal injection. The xenograft volumes were monitored timely. Immunohistochemical staining, RT-PCR and TUNEL assay were applied to investigate the related mechanisms that pGRIM-19-si-survivin inhibited tumor growth in vivo. RESULTS：Compared with psi-survivin and pGRIM-19 carried by attenuated Salmonella (control groups), the tumor volumes were reduced markedly in pGRIM-19-si-survivin plasmid group. The mean shrinkage rates were 2.36 and 3.02 times. pGRIM-19-si-survivin co-expression plasmid carried by attenuated Salmonella inhibited survivin expression but strengthened GRIM-19 expression obviously (P<0.05). The mRNA expression of apoptosis-related proteins such as Bcl-xL, Stat3, cyclin D1 and c-Myc was inhibited, and the vascular endothelial growth factor (VEGF) mRNA and Ki67 protein were also inhibited, but the caspase-3 mRNA expression was up-regulated (P<0.05) with significant cell apoptosis. CONCLUSION： pGRIM-19-si-survivin co-expression plasmid carried by human attenuated Salmonella inhibits the growth of prostate cancer subcutaneous xenografts in nude mice by promoting cell apoptosis and inhibiting prostatic cancer proliferation.     

Clonal Spread of Salmonella enterica Serovar Infantis in Serbia: Acquisition of Mutations in the Topoisomerase Genes gyrA and parC Leads to Increased Resistance to Fluoroquinolones

Quinolone‐resistant Salmonella Infantis (n = 64) isolated from human stool samples, food and poultry during the years 2006–2011 were analysed for their resistance phenotypes, macrorestriction patterns and molecular mechanisms of decreased susceptibility to fluoroquinolones. Minimum inhibitory concentrations (MICs) of nalidixic acid (NAL) and ciprofloxacin (CIP) were determined by the agar dilution procedure, and the susceptibility to additional antimicrobial agents was determined by the disc diffusion method. To assess the influence of enhanced efflux activity, MICs were determined in the presence and absence of the inhibitor PAβN. The results of pulsed‐field gel electrophoresis (PFGE) typing revealed that quinolone‐resistant S. Infantis in Serbia had similar or indistinguishable PFGE profiles, suggesting a clonal spread. All S. Infantis showed combined resistance to NAL and tetracycline, whereas multiple drug resistance to three or more antibiotic classes was rare (2 isolates of human origin). The MICs ranged between 512 and 1024 μg/mL for NAL and 0.125–2 μg/mL for CIP. A single‐point mutation in the gene gyrA leading to a Ser83→Tyr exchange was detected in all isolates, and a second exchange (Ser80→Arg) in the gene parC was only present in eight S. Infantis isolates exhibiting slightly higher MICs of CIP (2 μg/mL). The inhibitor PAβN decreased the MIC values of CIP by two dilution steps and of NAL by at minimum 3–6 dilution steps, indicating that enhanced efflux plays an important role in quinolone resistance in these isolates. The plasmid‐mediated genes qnr, aac(6′)‐lb‐cr and qepA were not detected by PCR assays.     

Phage and MLVA Typing of Salmonella Enteritidis Isolated from Layers and Humans in Belgium from 2000–2010, A Period in which Vaccination of Laying Hens was Introduced

The aim of the study was to characterize isolates of Salmonella enterica serovar Enteritidis (S. Enteritidis) obtained from humans and layer farms in Belgium collected during 2000–2010. Three periods were compared, namely (i) before implementation of vaccination (2000–2004), (ii) during voluntary vaccination (2005–2006) and (iii) during implementation of the national control program (NCP) for Salmonella including mandatory vaccination against S. Enteritidis (2007–2010). The characteristics compared across time periods were distributions of phage type and multiple‐locus variable number tandem‐repeat assay (MLVA). While PT4 and PT21 were predominantly isolated in Belgium in layers and humans before 2007, a significant reduction of those PTs was observed in both populations in the period 2007–2010. The relative proportion of PT4b, PT21c and PT6c was found to have increased considerably in the layer population since 2007. In the human population, PT8, PT1 and the group of ‘other’ PTs were more frequently isolated compared to the previous periods. When comparing the proportion of the predominant MLVA types Q2 and U2, no significant difference was found between the layer and human population in the three periods and between periods within each category (layer and human). A significant difference in isolate distribution among MLVA clusters I and II was found between human and layer isolates recovered during Period 3 and in the human population between Period 1 and 3. Results suggest that the association between S. Enteritidis in layers and the occurrence of the pathogen in humans changed since implementation of the NCP in 2007.     

沙门菌CRISPR的结构与功能研究进展

近年发现沙门菌中恒定地存在两个CRISPR位点,沙门菌利用它们不仅能抵御外源质粒和噬菌体的入侵,还能介导自身短期的表型变化及长期亚类的进化。同时由于其结构的多样性,使之广泛应用于沙门菌分型和进化的研究。论文简要综述了沙门菌CRISPR系统的基本结构、作用机制和功能及其在沙门菌分型和进化应用方面的研究进展。     

ε-聚赖氨酸复合物对大肠杆菌和沙门氏菌抑制作用研究

为了研究ε-聚赖氨酸对食品中常见G-菌的复合抑制效果,采用圆滤纸片法对大肠杆菌和沙门氏菌的抑菌效果进行了研究。试验结果表明:ε-聚赖氨酸对于大肠杆菌、沙门氏菌均能起到抑菌作用,随着ε-聚赖氨酸浓度的增大抑菌效果逐渐增强。不同浓度的ε-聚赖氨酸与0.1%双乙酸钠复合时抑菌圈虽然都有所增大,但协同增效作用不十分显著,而与0.2%曲酸复合试剂协同抑菌作用更为显著。     

Herd prevalence of Salmonella enterica infections in Danish slaughter pigs determined by microbiological testing

As a part of a nationwide programme to survey and control salmonella in pig herds, a microbiological survey of 1363 pig herds was performed in Denmark. A total of 13 468 slaughter pigs were examined at slaughter by culture of 5 g of caecal contents. Overall, 30 different serotypes of Salmonella enterica were isolated from 832 pigs (6.2%). The predominant serotype was S. Typhimurium, comprising 536 (64.4%) of the isolates. Four hundred and forty-eight isolates of S. Typhimurium were examined by phage typing, resulting in detection of 17 different phage types (definitive types, DT) with DT12 being the most frequent (49.1%).

Salmonella enterica was found in 302 herds (22.2%), S. Typhimurium was found in 61.1% of these. 279 (23.1%) large herds (producing more than 2600 slaughter pigs per year) were found to be salmonella positive compared with 23 (14.7 %) small herds (annual production of 500 to 550 slaughter pigs). Practical constraints in the study design did not allow for a firm conclusion on the interplay among herd size, geographical location and occurrence of salmonella.

In 284 of 302 infected herds (94.0%) only one serotype was detected. Infections with two different serovars were seen in 18 herds (6.0%).