Comparison of genetic and physiological properties of Salmonella enterica isolates from chickens reveals one major difference between serovar Kentucky and other serovars: response to acid.

For unknown reasons, Salmonella enterica Kentucky has become the serovar most frequently isolated from chickens and chicken carcasses in the United States. In an attempt to identify traits that may underlie this phenomenon, genetic and physiological features of 30 serovar Kentucky chicken isolates were compared with those of chicken isolates belonging to a range of other S. enterica serovars. Most of the well-known Salmonella virulence genes were detected in the serovar Kentucky isolates by PCR, but the cdtB, spvB, spvC, and pefA genes were not found. The serovar Kentucky isolates were as invasive as the non-Kentucky isolates in in vitro assays involving chicken embryo hepatocytes, but were less invasive than the Enteritidis, Mbandaki, and Typhimurium isolates when incubated with human HCT-8 cells. Statistical comparison of growth, biofilm formation, and stress survival data from the serovar Kentucky and those from the serovar Enteritidis, Hadar, Mbandaka, Senftenberg, Typhimurium, and Worthington isolates demonstrated either no differences or differences with only a few of the serovars; however, three data sets were different. These data sets included the OD(600) values of cultures grown in tryptic soy broth (TSB) adjusted to pH 5.5 with acetic acid and survival counts of cells grown in either TSB pH 7 or TSB adjusted to pH 5.5 with acetic acid and then transferred into TSB adjusted to pH 2.5 with HCl. Most notable was the log(10) reduction for acetic acid pre-exposed Kentucky isolates of 3.1 versus <1 log(10) for the other isolates upon transfer to pH 2.5. The connection, if any, between this acid response phenotype and the prevalence of the serovar Kentucky in poultry remains to be elucidated, but it is possible that slightly better growth in the presence of acetic acid in conjunction with not mounting a strong, energy-consuming acetic acid-induced adaptive acid response provides a small competitive advantage to this serovar in low acid environments such as the cecum where the pH is around 5.5.

Joerger RD ，Sartori CA ，Kniel KE 《-》

Gene expression analysis of Salmonella enterica Enteritidis Nal(R) and Salmonella enterica Kentucky 3795 exposed to HCl and acetic acid in rich medium.

In the United States, serovar Kentucky has become one of the most frequently isolated Salmonella enterica serovars from chickens. The reasons for this prevalence are not well understood. Phenotypic comparisons of poultry Salmonella isolates belonging to various serovars demonstrated that serovar Kentucky isolates differed from those of most other serovars in their response to acid. Microarray and qPCR analyses were performed with aerated exponentially growing poultry isolates, Salmonella enterica serovar Kentucky 3795 and Enteritidis Nal(R), exposed for 10 min to tryptic soy broth (TSB) adjusted to pH 4.5 with HCl and to pH 5.5 with HCl or acetic acid. Data obtained by microarray analysis indicated that more genes were up- or down-regulated in strain Kentucky 3795 than in Enteritidis Nal(R) under acidic conditions. Acid exposure in general caused up-regulation of energy metabolism genes and down-regulation of protein synthesis genes, particularly of ribosomal protein genes. Both strains appear to similarly utilize the lysine-based pH homeostasis system, as up-regulation of cadB was observed under the acidic conditions. Expression of regulatory genes (rpoS, fur, phoPQ) known to be involved in the acid response showed similar trends in both isolates. Differences between Kentucky 3795 and Enteritidis Nal(R) were observed with respect to the expression of the hdeB-like locus SEN1493 (potentially encoding a chaperone important to acid response), and some differences in the expression of other genes such as those involved in citrate utilization and motility were noted. It appears that the early stages of the transcriptional response to acid by isolates Kentucky 3795 and Enteritidis Nal(R) are similar, but differences exist in the scope and in some facets of the response. Possibly, the quantitative differences observed might lead to differences in protein levels that could explain the observed differences in the acid phenotype of serovar Kentucky and other Salmonella serovars.

Joerger RD ，Sartori C ，Frye JG ，Turpin JB ，Schmidt C ，McClelland M ，Porwollik S ... -  《-》

A comparative study of thermal and acid inactivation kinetics in fruit juices of Salmonella enterica serovar Typhimurium and Salmonella enterica serovar Senftenberg grown at acidic conditions.

Acid and heat inactivation in orange and apple juices of Salmonella enterica serovar Typhimurium Colección Española de Cultivos Tipo (i.e., Spanish Type Culture Collection) 443 (CECT 443) (Salmonella Typhimurium) and S. enterica serovar Senftenberg CECT 4384 (Salmonella Senftenberg) grown in buffered brain heart infusion (pH 7.0) and acidified brain heart infusion up to pH 4.5 with acetic, citric, lactic, and hydrochloric acids was evaluated. Acid adaptation induced an adaptive response that increased the subsequent resistance to extreme pH conditions (pH 2.5) and to heat, although the magnitude of these responses differed between the two isolates and fruit juices. The acid resistance in orange juice for acid-adapted cells (D-values of 28.3-34.5 min for Salmonella Senftenberg and 30.0-39.2 min for Salmonella Typhimurium) resulted to be about two to three times higher than that corresponding to non-acid-adapted cells. In apple juice, acid-adapted Salmonella Senftenberg cells survived better than those of Salmonella Typhimurium, obtaining mean D-values of 114.8 +/- 12.3 and 41.9 +/- 2.5 min, respectively. The thermotolerance of non-acid-adapted Salmonella Typhimurium in orange (D(58)-value: 0.028 min) and apple juices (D(58)-value: 0.10 min) was approximately double for acid-adapted cells. This cross-protection to heat was more strongly expressed in Salmonella Senftenberg. D(58)-values obtained for non-acid-adapted cells in orange (0.11 min) and apple juices (0.19 min) increased approximately 10 and 5 times, respectively, after their growth in acidified media. The conditions prevailing during bacterial growth and heat treatment did not significantly influence the z-values observed (6.0 +/- 0.3 degrees C for Salmonella Typhimurium and 7.0 +/- 0.3 degrees C for Salmonella Senftenberg). The enhanced acid resistance found for both isolates could enable them to survive for prolonged time periods in the gastrointestinal tract, increasing the risk of illness. Further, it should be taken into account that microbial growth in acidified media also induces a cross-protection response against heat that should also be considered for the design of pasteurization processes for acid foods.

Alvarez-Ordóñez A ，Fernández A ，Bernardo A ，López M ... -  《-》

Adaptive acid tolerance response in Salmonella enterica serovar Typhimurium and Salmonella enterica serovar Typhi.

The survival of bacteria in various environments depends on a number of protective responses including acid tolerance response (ATR). In this study, ATR phenomenon was compared in Salmonella enterica serovar Typhi 6 and Salmonella enterica serovar Typhimurium 98 under different culture conditions. Survival of the adapted culture (pre-acid shocked to pH 5.5) was significantly better (p < 0.05) as compared to control, unadapted culture after acid shock at pH 3.3. However, the ATR varied with the serovar, incubation temperature and the growth medium used (all p-values < 0.05). S. Typhi 6 failed to grow in pH 3.3 at 45 degrees C. The addition of tetracycline or chloramphenicol (1.0 microg ml(-1)) to adapted cultures during or after acid shock (pH 3.3) had no effect on ATR expression. In S. Typhimurium 98, growth was increased by 10% or greater in adapted culture (when grown at pH 3.3) as compared to growth observed with an unadapted culture (when grown at pH 7.3) on transfer to fresh growth medium at pH 7.3. A poor ATR observed in non-growing S. Typhimurium 98 suspensions clearly showed that ATR is an energy-consuming process. Storage of S. Typhimurium 98 cultures in pH 4.5 nutrient broth at 4 degrees C demonstrated that prolonged exposure to acidic conditions is more detrimental in comparison to the cultures stored at pH 7.3 at this temperature.

Tiwari RP ，Sachdeva N ，Hoondal GS ，Grewal JS ... -  《JOURNAL OF BASIC MICROBIOLOGY》

Comparative analysis of acid resistance in Listeria monocytogenes and Salmonella enterica strains before and after exposure to poultry decontaminants. Role of the glutamate decarboxylase (GAD) system.

Data on the ability of chemical poultry decontaminants to induce an acid stress response in pathogenic bacteria are lacking. This study was undertaken in order to compare the survival rates in acid broths of Listeria monocytogenes and Salmonella enterica strains, both exposed to and not exposed to decontaminants. The contribution of the glutamate decarboxylase (GAD) acid resistance system to the survival of bacteria in acid media was also examined. Four strains (L. monocytogenes serovar 1/2, L. monocytogenes serovar 4b, S. enterica serotype Typhymurium and S. enterica serotype Enteritidis) were tested before (control) and after exposure to trisodium phosphate, acidified sodium chlorite, citric acid, chlorine dioxide and peroxyacids (strains were repeatedly passed through media containing increasing concentrations of a compound). Stationary-phase cells (10(8) cfu/ml) were inoculated into tryptic soy broth (TSB) acidified with citric acid (pH 2.7 and 5.0) with or without glutamate (10 mM) added, and incubated at 37 degrees C for 15 min. Survival percentages (calculated from viable colonies) varied from 2.47 +/- 0.67% to 91.93 +/- 5.83%. L. monocytogenes cells previously exposed to acid decontaminants (citric acid and peroxyacids) showed, when placed in acid TSB, a higher (P < 0.05) percentage of survival (average 38.80 +/- 30.52%) than control and pre-exposed to non-acidic decontaminants strains (22.82 +/- 23.80%). Similar (P > 0.05) survival percentages were observed in previously exposed to different decontaminants and control Salmonella strains. The GAD acid resistance system did not apparently play any role in the survival of L. monocytogenes or S. enterica at a low pH. This study demonstrates for the first time that prior exposure to acidic poultry decontaminants increases the percentage of survival of L. monocytogenes exposed to severe acid stress. These results have important implications for the meat industry when considering which decontaminant treatment to adopt.

Alonso-Hernando A ，Alonso-Calleja C ，Capita R 《-》