Salmonella enterica serovar Pullorum requires the Salmonella pathogenicity island 2 type III secretion system for virulence and carriage in the chicken.

Functional mutations were made in the type III secretion systems encoded by Salmonella pathogenicity island 1 (SPI 1) and Salmonella pathogenicity island 2 (SPI 2) of Salmonella enterica serovar Pullorum, the cause of pullorum disease in poultry. Their role in cell invasion in vitro, and in virulence in vivo was determined. The SPI 1 mutant showed decreased invasiveness for chicken cells but was capable of causing disease in orally infected 1-day-old chicks, although it showed some reduction in virulence. The SPI 2 mutant showed no reduction in invasiveness, but was fully attenuated for virulence in 1-day-old chicks, and was not detected following oral infection in 1-week-old chickens. Following intravenous infection, the SPI 2 mutant was also attenuated and cleared more rapidly than the parent strain. This indicates that S. Pullorum requires SPI 2 for virulence and persistence but SPI 1 appears to contribute to, but is not essential for, the virulence of S. Pullorum.

Wigley P ，Jones MA ，Barrow PA 《AVIAN PATHOLOGY》

The Salmonella pathogenicity island 1 and Salmonella pathogenicity island 2 type III secretion systems play a major role in pathogenesis of systemic disease and gastrointestinal tract colonization of Salmonella enterica serovar Typhimurium in the chicken.

Salmonella enterica serovar Typhimurium infection of chickens is a major public and animal health problem. In young chicks, S. Typhimurium infection results in severe systemic infection; in older chicks, infection results in prolonged gastrointestinal tract colonization. Here we determined the role of the Salmonella pathogenicity island 1 (SPI-1) and Salmonella pathogenicity island 2 (SPI-2) type III secretion systems in systemic infection and gastrointestinal tract colonization of the chicken though experimental infection of chicks with a S. Typhimurium strain with mutations in the genes encoding the secretion system machinery of SPI-1 (spaS) and SPI-2 (ssaU) that prevent secretion of effector proteins. In 1-day-old chicks, mutation of SPI-2 lead to a decrease in both systemic bacterial numbers and pathology, although no difference in gastrointestinal numbers was observed. Mutation of SPI-1 had little effect in 1-day old chicks. In 1-week-old animals the SPI-2 mutants could not be detected systemically and colonized the gastrointestinal tract only in low numbers in comparison with the parent strain, and was cleared in 1 week. The SPI-1 mutant showed greatly reduced levels of systemic infection, and colonized the gastrointestinal tract at a lower level than the parent strain. The findings show that the SPI-2 type III secretion system is required for systemic S. Typhimurium infection in both infection models, and that it plays a significant role in gastrointestinal colonization. The SPI-1 system is involved in both systemic infection and gastrointestinal colonization, but does not appear absolutely essential for either infection process.

Jones MA ，Hulme SD ，Barrow PA ，Wigley P ... -  《AVIAN PATHOLOGY》

Salmonella enterica serovar Gallinarum requires the Salmonella pathogenicity island 2 type III secretion system but not the Salmonella pathogenicity island 1 type III secretion system for virulence in chickens.

Salmonella enterica serovar Gallinarum is a host-specific serotype that causes the severe systemic disease fowl typhoid in domestic poultry and a narrow range of other avian species but rarely causes disease in mammalian hosts. Specificity of the disease is primarily at the level of the reticuloendothelial system, but few virulence factors have been described other than the requirement for an 85-kb virulence plasmid. In this work, by making functional mutations in the type III secretion systems (TTSS) encoded by Salmonella pathogenicity island 1 (SPI-1) and SPI-2, we investigated the role of these pathogenicity islands in interactions between Salmonella serovar Gallinarum and avian cells in vitro and the role of these pathogenicity islands in virulence in chickens. The SPI-1 mutant showed decreased invasiveness into avian cells in vitro but was unaffected in its ability to persist within chicken macrophages. In contrast the SPI-2 mutant was fully invasive in nonphagocytic cells but failed to persist in macrophages. In chicken infections the SPI-2 mutant was attenuated while the SPI-1 mutant showed full virulence. In oral infections the SPI-2 mutant was not observed in the spleen or liver, and following intravenous inoculation it was cleared rapidly from these sites. SPI-2 function is required by Salmonella serovar Gallinarum for virulence, primarily through promoting survival within macrophages allowing multiplication within the reticuloendothelial system, but this does not preclude the involvement of SPI-2 in uptake from the gut to the spleen and liver. SPI-1 appears to have little effect on virulence and survival of Salmonella serovar Gallinarum in the host.

Jones MA ，Wigley P ，Page KL ，Hulme SD ，Barrow PA ... -  《-》

Identification of a new Salmonella enterica serovar Enteritidis locus involved in cell invasion and in the colonisation of chicks.

Poultry products contaminated with Salmonella enterica serovar Enteritidis are a major cause of foodborne disease in industrialized countries. Knowledge of how poultry is colonised is essential for reducing contamination of these products. We have characterized the bacterial yfg-eng locus involved in chicken colonisation. Its sequencing revealed four open reading frames (ORF), yfgM, yfgL, engA and yfgJ, all transcribed in the same orientation. An yfgL mutant of S. Enteritidis colonised the caeca (P < 0.05) and the spleens (P < 0.01) of one-day-old chicks subnormally 2 and 5 days after oral inoculation. This lower virulence was correlated with reduced secretion of the SPI-1 and flagellar proteins in the yfgL mutant compared to the wild-type strain. Consistent with this, the S. Enteritidis yfgL mutant was less motile than the wild type and fewer invaded enterocytes (P < 0.05) and avian HD11 macrophages (P < 0.001). All these defects could be partially overcome by inserting the yfg-eng locus into the mutant on a recombinant plasmid.

Amy M ，Velge P ，Senocq D ，Bottreau E ，Mompart F ，Virlogeux-Payant I ... -  《RESEARCH IN MICROBIOLOGY》

[Construction and characterization of Salmonella Pathogenicity Island 2 (SPI-2) deletion mutant of Salmonella Pullorum S06004].

To research the pathogenicity of Salmonella Pathogenicity Island 2 (SPI-2) deletion mutant of Salmonella Pullorum and preliminary explore the feasibility of developing safe attenuated Salmonella Pullorum candidate vaccine strain. The SPI-2 (-40 kb) deletion mutant of Salmonella Pullorum S06004 was constructed using the λ-red recombinant system. Then the biological characteristics such as growth rate, biochemical properties, genetic stability and virulence were evaluated between the deletion mutant strain S06004ΔSPI2 and its parent strain S06004. S06004ΔSPI2 was successfully constructed. The growth rate and biochemical properties of S06004ΔSPI2 were consistent with those of its parent strain S06004. The mutant was stable with the deletion of SPI-2. Chicken lethal test showed that the LD50 of S06004ΔSPI2 was 252 times higher than the parent strain S06004. The virulence of S06004ΔSPI2 was obviously attenuated. This study provided basic data for further study of the functions of SPI-2, and implied its potential to develop attenuated Salmonella vaccine.

Yin J ，Wu Y ，Lin Z ，Wang X ，Hu Y ，Li Q ，Geng S ，Jiao X ... -  《-》