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Role of Salmonella enterica subspecies enterica serovar Enteritidis pathogenicity island-2 in chickensWisner, Amanda Lynn Stacy 02 August 2011 (has links)
Salmonella enterica subspecies enterica serovar Enteritidis (S. Enteritidis) has been identified as a significant cause of salmonellosis in humans. Salmonella pathogenicity islands 1 and 2 (SPI-1 and SPI-2) each encode a specialized type III secretion system (T3SS) that enables Salmonella to manipulate host cells at various stages of the invasion/infection process. The SPI-2 T3SS has been identified as vital for survival and replication of S. Typhimurium and S. Enteritidis in mouse macrophages, as well as full virulence in mice. In order to test the ability of SE SPI-2 mutants to survive in vivo we used a chicken isolate of SE (Sal18). In one study, we orally co-challenged 35-day-old specific pathogen free (SPF) chickens with two bacterial strains per group. The control group received two versions of the wild-type (WT) strain Sal18: Sal18 attTn7::tet and Sal18 attTn7::cat, while the other two groups received the WT strain (Sal18 attTn7::tet) and one of two mutant strains (Sal18 attTn7::cat ÄspaSÄssaU or Sal18 ÄSPI-1ÄSPI-2::cat). From this study we conclude that S. Enteritidis deficient in the SPI-1 and SPI-2 systems are out-competed by the WT strain. In a second study, groups of SPF chickens were challenged at 1 week of age with four different strains; a WT strain and three other strains missing either one or both of the SPI-1 and SPI-2 regions. On days 1 and 2 post-challenge (PC) we observed a reduced systemic spread of the SPI-2 mutants, but by day 3 the mutants systemic distribution levels matched that of the WT strain. Based on these two studies, we conclude that the SPI-2 T3SS facilitates invasion and systemic spread of S. Enteritidis in chickens, but alternative mechanisms for these processes appear to exist.
Several structural components of the T3SSs encoded by SPI-1 and SPI-2 are exposed to the hosts immune system prior to/during the infection/invasion process, making them potential vaccine candidates. Several of these candidates genes were cloned, the proteins overproduced, purified, and formulated as vaccines for use in further studies. SPI-2 T3SS proteins used for vaccine studies included the secretin, SsaC, the needle, SsaG, the filament, SseB, and a part of the translocon, SseD, as well as a number of effectors, SseI, SseL, SifA, and SifB. The first vaccine study involved vaccination of SPF chickens with SseB and SseD, followed by challenge with the WT S. Enteritidis strain Sal18. Additional studies evaluated whether hens vaccinated with SPI-2 T3SS structural or effector components could mount a significant humoral immune response (as measured by serum immunoglobulin Y [IgY] titres), whether these antibodies could be transferred to progeny (as measured by egg yolk IgY titres), and whether vaccinates and progeny of vaccinates could be protected against challenge with the WT S. Enteritidis strain Sal8. The results of our studies show that vaccinated chickens do produce high levels of SPI-2 T3SS specific serum IgY that they are able to transfer to their progeny. It was demonstrated that vaccinates and progeny of vaccinates had lower overall countable recovered SE per bird in most situations.
In order to better identify the role of the SPI-2 T3SS in chickens, we used the well-known gentamicin protection assay with activated HD11 cells. HD11 cells are a macrophage-like chicken cell line that can be stimulated with phorbol 12-myristate 13-acetate (PMA) to exhibit more macrophage-like morphology and greater production of reactive oxygen species (ROS). Activated HD11 cells were infected with a WT S. Typhimurium strain, a SPI-2 mutant S. Typhimurium strain, a WT S. Enteritidis strain, a SPI-2 mutant S. Enteritidis strain, or a non-pathogenic Escherichia coli (E. coli) strain. SPI-2 mutant strains were found to survive as well as their parent strain at all time points post-infection (PI) up to 24 h PI, while the E. coli strain was no longer recoverable by 3 h PI. We can conclude from these observations that the SPI-2 T3SS is not important for survival of Salmonella in the activated macrophage-like HD11 cell line, and that Salmonella must employ other mechanisms for survival in this environment as E. coli is effectively eliminated.
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Evaluation of Alternative Cooking and Cooling Procedures for Large, Intact Meat Products to Achieve Lethality and Stabilization Microbiological Performance StandardsHaneklaus, Ashley 16 January 2010 (has links)
This study was conducted to determine if alternative heating times and slower
cooling times, other than those defined by FSIS, could be utilized and still comply with
FSIS performance standards. Large (10.43 to 12.25 kg), cured bone-in hams (n = 190)
and large (greater than or equal to 9.07 kg), uncured beef inside rounds (n = 180) were utilized in a two-phase
study. Phase 1 of the study investigated the effect of alternative lethality parameters on
toxin production of Staphylococcus aureus and log reduction of Salmonella
Typhimurium and coliforms. Both the hams and roast beef were subjected to 1 of 10
treatments defined by varying final internal product temperatures (48.9 degrees C, 54.4 degrees C,
60.0 degrees C, 65.6 degrees C, or 71.1 degrees C) and smokehouse relative humidities (50% or 90%). Phase 2
investigated the effect of alternative stabilization parameters on log growth of
Clostridium perfringens. Stabilization treatments extended the times taken to reduce
internal product temperature from 54.4 degrees C to 26.7 degrees C and from 26.7 degrees C to 7.2 degrees C (ham) or
4.5 degrees C (beef), independently. Further, a control treatment following current FSIS,
Appendix B guidelines was conducted for ham, and a "worst case" scenario was
assessed for both products. The "worst case" treatment evaluated the effects of cooling products at room temperature (approximately 22.8 degrees C) in place of normal cooling
procedures in a temperature controlled environment. Results of the study showed at
least a 6.5-log10 reduction in S. Typhimurium across all lethality treatments for both
products. Further, coliform counts also were reduced significantly, and S. aureus toxin
kits returned negative results for toxin production for all treatments of ham and roast
beef. Stabilization showed less than 1-log growth of C. perfringens for any treatment,
with the exception of the "worst case" scenario for roast beef. As expected, > 1 log
growth of C. perfringens was found for uncured roast beef maintained at room
temperature for cooling. This study supports that there are multiple time and
temperature combinations, other than those currently provided by FSIS, which may be
utilized for cooking and cooling large roast beef and bone-in ham products while still
meeting FSIS lethality and stabilization microbiological performance standards.
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Post-transcriptional regulation of rpoS and HemA in salmonellaJones, Amy Madeline. January 2009 (has links)
Thesis (Ph. D.)--West Virginia University, 2009. / Title from document title page. Document formatted into pages; contains vii, 104 p. : ill. Includes abstract. Includes bibliographical references.
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The ecology and evolution of antimicrobial resistance in asymptomatic Salmonella enterica /Guimond-Peron, Gabriel. January 2006 (has links)
Infections caused by resistant pathogens fail to respond to treatment, resulting in increased costs due to prolonged illness and hospitalization. Determining the extent of resistance in animal populations is thus of great importance to public health. In this work, we first showed that asymptomatic populations of Salmonella in pigs present greater genotypic and phenotypic diversity than disease-associated populations. Second, we identified a clonal population structure associated with asymptomatic Salmonella found in the Canadian swine industry and we confirmed that food-producing pigs are a significant reservoir of Salmonella enterica, more particularly the clinically important serotype Typhimurium DT104. Finally, we identified the possible independent evolution of multidrug-resistance in serotypes Typhimurium, Derby and Heidelberg. Our work on asymptomatic Salmonella enterica stresses the importance of linking ecology and evolutionary biology to public health in order to understand and predict the response of pathogenic bacteria to selective pressure imposed by host immunity, whether naturally or artificially induced.
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The combined effect of MAP and other barriers on the growth of Salmonella enteritidis in packaged chicken thighs under various storage conditions /Al-Zenki, Sameer F. January 1996 (has links)
Salmonella enteritidis has recently emerged as a potential pathogen in poultry products. The growth of S. enteritidis in poultry is affected by several factors such as storage temperature, pH, water activity, modified atmosphere and the presence of preservatives. All of these factors may act alone or in combination with each other resulting in a synergistic, antimicrobial effect. / In this research, initial storage studies were done to determine the effect of various atmospheres (air, vacuum, oxygen absorbent and gas packaging) on the microbial changes of packaged chicken thighs followed by challenge studies with a strain of S. enteritidis$ sp{ rm{NAST}}$. Chicken thighs were packaged in Cryovac bags and stored at 4 and 12$ sp circ$C for up to 28d. Changes in headspace gas composition, pH, drip loss, color and odor were monitored at each sampling day. / The effect of various packaging treatments, dipping solutions (chitosan (0.2%w/v) and potassium sorbate (0.2%w/v)) and low dose irradiation (1.5 & 3.0 kGy) on the growth of S. enteritidis$ sp{ rm NAST}$ and on the shelf-life of chicken thighs stored at 4 and 12$ sp circ$C was also investigated. (Abstract shortened by UMI.)
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Importance of wild birds in the spread of SalmonellaPalmgren, Helena January 2002 (has links)
Salmonella is one of the most important enteropathogenic bacteria. It is responsible for about 5000 reported cases of human gastroenteritis each year in Sweden. Salmonellosis is a zoonotic disease, and the bacterium has the ability to infect a variety of both domestic and wild animal species. In studies of Swedish wild bird populations, we found that Black-headed gull may be the main reservoir for Salmonella in birds, and that Salmonella infection is expressed as carriage with no obvious disease manifestations. Black-headed gull is a migratory bird and can transport strains of Salmonella with virulence traits like antibiotic resistance, from sources outside Sweden. Genetic molecular methods, PFGE and IS200, also demonstrate that Black-headed gull play a role in the transmission chain of Salmonella in Sweden. In a study of the Swedish Peregrine Falcon population, Salmonella amager and Campylobacter jejuni were found. There were indications, based on serotyping of Salmonella and genetical typing by PFGE of Campylobacter that these isolates were transmitsted to the falcons from a human or domestic animal source. This bird of prey has sparse contact with humans but may be infected by Salmonella of human origin by feeding on other birds, like gull. Salmonella was found in penguins, albatrosses and mainly in seals in a study in Antarctica. Several features of the Salmonella serotypes found indicate a human source for Salmonella infection in these animals, and also a spread of Salmonella within and between animal species in Antarctica. / <p>Diss. (sammanfattning) Umeå : Umeå universitet, 2002</p> / digitalisering@umu
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The Mutagenic Activity of High-Energy Explosives; Contaminants of Concern at Military Training SitesMcAllister, Jennifer E. 24 August 2011 (has links)
The genotoxicity of energetic compounds (i.e., explosives) that are known to be present in contaminated soils at military training sites has not been extensively investigated. Thus, the Salmonella mutagenicity and Muta(TM)Mouse assays were employed as in vitro assays to examine the mutagenic activity of twelve explosive compounds, as well as three soil samples from Canadian Forces Base Petawawa. Salmonella analyses employed strains TA98 (frameshift mutations) and TA100 (base-pair substitution mutations), as well as the metabolically-enhanced YG1041 (TA98 background) and YG1042 (TA100 background), with and without exogenous metabolic activation (S9). For Salmonella analyses, the results indicate that ten of the explosive compounds were mutagenic, and consistently elicited direct-acting, base-pair substitution activity. All three soil samples were also observed to be mutagenic, eliciting direct-acting, frameshift activity. Mutagenic potencies were significantly higher on the metabolically-enhanced strains for all compounds and soil samples. For Muta(TM)Mouse analyses on FE1 cells, the results indicate that the majority of explosive compounds did not exhibit mutagenic activity. All three soil samples elicited significant positive responses (PET 1 and PET 3 without S9, and PET 2 with S9), and although there is some evidence of a concentration-related trend, the responses were weak. Correspondence of the mutagenic activity observed with the two assay systems, for both the explosive compounds and soil samples, was negligible. The differential response is likely due to differences in metabolic capacity between the two assay systems. Furthermore, it is likely that there are unidentified compounds present in these soil samples that are, at least in part, responsible for the observed mutagenic activity. Additional testing of other explosive compounds, as well as soil samples from other military training sites, using a variety of in vitro and in vivo assays, is warranted in order to reliably estimate mutagenic hazard and subsequently assess risk to human health.
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Construction of Salmonella vaccines / David HoneHone, David January 1988 (has links)
Bibliography: leaves 126-171 / xiv, 171 leaves : ill ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--University of Adelaide, Dept. of Microbiology and Immunology, 1988
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Characterization of c-di-GMP signalling in Salmonella typhimurium /Simm, Roger, January 2007 (has links)
Diss. (sammanfattning) Stockholm : Karolinska institutet, 2007. / Härtill 4 uppsatser.
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Mechanisms of adaptation to the fitness cost of antibiotic resistance /Paulander, Wilhelm, January 2007 (has links)
Diss. (sammanfattning) Stockholm : Karolinska institutet, 2007. / Härtill 3 uppsatser.
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