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PubMed2001

Salmonella pathogenicity island 2 influences both systemic salmonellosis and Salmonella-induced enteritis in calves.

We have used signature-tagged mutagenesis to identify mutants of the host-specific Salmonella enterica serotype Dublin which were avirulent in calves and/or BALB/c mice. A mutant with a transposon insertion in the sseD gene of Salmonella pathogenicity island 2 (SPI-2), which encodes a putative secreted effector protein, was identified. This mutant was recovered from the bovine host but not from the murine host following infection with a pool of serotype Dublin mutants. However, a pure inoculum of the sseD mutant was subsequently shown to be attenuated in calves following infection either by the intravenous route or by the oral route. The sseD mutant was fully invasive for bovine intestinal mucosa but was subsequently unable to proliferate to the same numbers as the parental strain in vivo. Both the sseD mutant and a second SPI-2 mutant, with a transposon insertion in the ssaT gene, induced significantly weaker secretory and inflammatory responses in bovine ligated ileal loops than did the parental strain. These results demonstrate that SPI-2 is required by serotype Dublin for the induction of both systemic and enteric salmonellosis in calves.

Animals↗
PubMed2004

Role of the Salmonella pathogenicity island 1 effector proteins SipA, SopB, SopE, and SopE2 in Salmonella enterica subspecies 1 serovar Typhimurium colitis in streptomycin-pretreated mice.

Salmonella enterica subspecies 1 serovar Typhimurium (serovar Typhimurium) induces enterocolitis in humans and cattle. The mechanisms of enteric salmonellosis have been studied most extensively in calf infection models. The previous studies established that effector protein translocation into host cells via the Salmonella pathogenicity island 1 (SPI-1) type III secretion system (TTSS) is of central importance in serovar Typhimurium enterocolitis. We recently found that orally streptomycin-pretreated mice provide an alternative model for serovar Typhimurium colitis. In this model the SPI-1 TTSS also plays a key role in the elicitation of intestinal inflammation. However, whether intestinal inflammation in calves and intestinal inflammation in streptomycin-pretreated mice are induced by the same SPI-1 effector proteins is still unclear. Therefore, we analyzed the role of the SPI-1 effector proteins SopB/SigD, SopE, SopE2, and SipA/SspA in elicitation of intestinal inflammation in the murine model. We found that sipA, sopE, and, to a lesser degree, sopE2 contribute to murine colitis, but we could not assign an inflammation phenotype to sopB. These findings are in line with previous studies performed with orally infected calves. Extending these observations, we demonstrated that in addition to SipA, SopE and SopE2 can induce intestinal inflammation independent of each other and in the absence of SopB. In conclusion, our data corroborate the finding that streptomycin-pretreated mice provide a useful model for studying the molecular mechanisms of serovar Typhimurium colitis and are an important starting point for analysis of the molecular events triggered by SopE, SopE2, and SipA in vivo.

Animals↗
PubMed2005

Salmonella-induced filament formation is a dynamic phenotype induced by rapidly replicating Salmonella enterica serovar typhimurium in epithelial cells.

Salmonella enterica serovar Typhimurium has the fascinating ability to form tubular structures known as Salmonella-induced filaments (Sifs) in host cells. Here, we show that the prevalence of the Sif phenotype in HeLa cells is affected by host cell density, growth, and the multiplicity of infection. Sif formation was observed in cells that displayed rapid intracellular bacterial replication and was found to be dynamic, being maximal 8 to 10 h postinfection and declining thereafter. The virulence factors SpvB and SseJ were found to negatively modulate Sif formation. Our findings demonstrate the complex and dynamic nature of the Sif phenotype.

Cell Division↗
PubMed1994

Inactivation of mismatch repair overcomes the barrier to transduction between Salmonella typhimurium and Salmonella typhi.

P22 transduction of chromosomal genes from Salmonella typhimurium into Salmonella typhi occurs at a low frequency. Transduction of plasmids from S. typhimurium into S. typhi occurs at a frequency similar to that between S. typhimurium strains, indicating that the barrier to transduction of chromosomal genes is not due to an inability of P22 to inject DNA into S. typhi or a restriction endonuclease that rapidly degrades foreign DNA. Furthermore, transduction of mutS and mutL derivatives of S. typhi with chromosomal genes from S. typhimurium occurs efficiently. These results indicate that the transduction barrier is due to activity of the recipient mismatch repair system, which senses sequence divergence and disrupts heteroduplexes in favor of recipient sequences. Inactivation of the mismatch repair system allows P22 transduction to be used as an effective tool for constructing S. typhi-S. typhimurium hybrids.

DNA Repair↗
PubMed1998

Identification of a new site for ferrichrome transport by comparison of the FhuA proteins of Escherichia coli, Salmonella paratyphi B, Salmonella typhimurium, and Pantoea agglomerans.

The fhuA genes of Salmonella paratyphi B, Salmonella typhimurium, and Pantoea agglomerans were sequenced and compared with the known fhuA sequence of Escherichia coli. The highly similar FhuA proteins displayed the largest difference in the predicted gating loop, which in E. coli controls the permeability of the FhuA channel and serves as the principal binding site for the phages T1, T5, and phi80. All the FhuA proteins contained the region in the gating loops required in E. coli for ferrichrome and albomycin transport. The three subdomains required for phage binding were contained in the gating loop of S. paratyphi B which is infected by the E. coli phages, whereas two of the subdomains were deleted in S. typhimurium and P. agglomerans which are resistant to the E. coli phages. Small deletions in a surface loop adjacent to the gating loop, residues 236 to 243 and 236 to 248, inactivated E. coli FhuA with regard to transport of ferrichrome and albomycin, but sensitivity to T1 and T5 was fully retained and sensitivity to phi80 and colicin M was reduced 10-fold. Full-size FhuA hybrid proteins of S. paratyphi B and S. typhimurium displayed S. paratyphi B FhuA activity when the hybrids contained two-thirds of either the N- or the C-terminal portions of S. paratyphi B and displayed S. typhimurium FhuA activity to phage ES18 when the hybrid contained two-thirds of the N-terminal region of the S. typhimurium FhuA. The central segment of the S. paratyphi B FhuA flanked on both sides by S. typhimurium FhuA regions conferred full sensitivity only to phage T5. The data support the essential role of the gating loop for the transport of ferrichrome and albomycin, identified an additional loop for ferrichrome and albomycin uptake, and suggest that several segments and their proper conformation, determined by the entire FhuA protein, contribute to the multiple FhuA activities.

Amino Acid Sequence↗
PubMed1998

Gene transfer between related bacteria by electrotransformation: mapping Salmonella typhi genes in Salmonella typhimurium.

Transfer of newly isolated mutations into a fresh background is an essential step of genetic analysis and strain construction. Gene transfer is hampered in Salmonella typhi and in other pathogenic bacteria by the lack of a generalized transduction system. We show here that this problem can be partially circumvented by using electrotransformation as a means for delivering S. typhi DNA into suitable S. typhi or Salmonella typhimurium recipients. Transferred DNA can recombine with the homologous region in the host chromosome. In one application of the method, mutations isolated in S. typhi were genetically mapped in S. typhimurium.

Chromosome Mapping↗
PubMed2002

Inversions over the terminus region in Salmonella and Escherichia coli: IS200s as the sites of homologous recombination inverting the chromosome of Salmonella enterica serovar typhi.

Genomic rearrangements (duplications and inversions) in enteric bacteria such as Salmonella enterica serovar Typhimurium LT2 and Escherichia coli K12 are frequent (10(-3) to 10(-5)) in culture, but in wild-type strains these genomic rearrangements seldom survive. However, inversions commonly survive in the terminus of replication (TER) region, where bidirectional DNA replication terminates; nucleotide sequences from S. enterica serovar Typhimurium LT2, S. enterica serovar Typhi CT18, E. coli K12, and E. coli O157:H7 revealed genomic inversions spanning the TER region. Assuming that S. enterica serovar Typhimurium LT2 represents the ancestral genome structure, we found an inversion of 556 kb in serovar Typhi CT18 between two of the 25 IS200 elements and an inversion of about 700 kb in E. coli K12 and E. coli O157:H7. In addition, there is another inversion of 500 kb in E. coli O157:H7 compared with E. coli K12. PCR analysis confirmed that all S. enterica serovar Typhi strains tested, but not strains of other Salmonella serovars, have an inversion at the exact site of the IS200 insertions. We conclude that inversions of the TER region survive because they do not significantly change replication balance or because they are part of the compensating mechanisms to regain chromosome balance after it is disrupted by insertions, deletions, or other inversions.

Chromosome Inversion↗
PubMed2003

Role of the Salmonella pathogenicity island 1 (SPI-1) protein InvB in type III secretion of SopE and SopE2, two Salmonella effector proteins encoded outside of SPI-1.

Salmonella enterica subspecies 1 serovar Typhimurium encodes a type III secretion system (TTSS) within Salmonella pathogenicity island 1 (SPI-1). This TTSS injects effector proteins into host cells to trigger invasion and inflammatory responses. Effector proteins are recognized by the TTSS via signals encoded in their N termini. Specific chaperones can be involved in this process. The chaperones InvB, SicA, and SicP are encoded in SPI-1 and are required for transport of SPI-1-encoded effectors. Several key effector proteins, like SopE and SopE2, are located outside of SPI-1 but are secreted in an SPI-1-dependent manner. It has not been clear how these effector proteins are recognized by the SPI-1 TTSS. Using pull-down and coimmunoprecipitation assays, we found that SopE is copurified with InvB, the known chaperone for the SPI-1-encoded effector protein Sip/SspA. We also found that InvB is required for secretion and translocation of SopE and SopE2 and for stabilization of SopE2 in the bacterial cytosol. Our data demonstrate that effector proteins encoded within and outside of SPI-1 use the same chaperone for secretion via the SPI-1 TTSS.

Animals↗
PubMed1979

Serological response of chickens to Salmonella thompson and Salmonella pullorum infections.

Chickens were experimentally infected with Salmonella thompson (serogroup C, paratyphoid) and Salmonella pullorum (serogroup D). Five serological methods and one cultural method were used in detecting the infections. The microantiglobulin test was superior to all other methods for detection of paratyphoid (S. THOMPSON) infection and was followed in efficacy by the microagglutination test, rapid serum plate test, cloacal swab culture, macroscopic tube agglutination test, and rapid whole-blood test, in that order. Birds infected with S. pullorum showed much higher agglutinin titers than the birds infected with paratyphoid. The microagglutination and microantiglobulin tests were not significantly different for detection of pullorum infection and were followed in efficacy by the rapid serum plate, macroscopic tube agglutination, rapid whole-blood, and cloacal swab culture tests, in that order. The cloacal swab culture test was totally inadequate for the detection of pullorum infection.

Agglutination Tests↗
PubMed1979

Salmonella bacteriophage glycanases: endorhamnosidases of Salmonella typhimurium bacteriophages.

Twelve bacteriphages lysing only smooth Salmonella typhimurium strains were shown to have similar morphology--an icosahedric head to which a short, noncontractile tail carrying six spikes was attached. All phages degraded their lipopolysaccharide (LPS) receptors as shown by their ability to cleave off [14C]galactosyl-containing oligosaccharides from S. typhimurium cells labeled in their LPS. The oligosaccharides inhibited the alpha-D-galactosyl-specific Bandeiraea simplicifolia lectin agglutination of human type B erythrocytes, indicating that all 12 phage glycanases were of endorhamnosidase specificity, i.e., hydrolyzed the alpha-L-rhamnopyranosyl-(1 leads to 3)-D-galactopyranosyl linkage in the S. typhimurium O-polysaccharide chain. Two of the phages, 28B and 36, were studied in more detail. Whereas the phage 28B glycanase hydrolyzed the S. typhimurium LPS into dodeca- and octasaccharides, the phage 36 glycanase in addition cleaved off tetrasaccharides. Both phage enzymes hydrolyzed the O-polysaccharide chains of LPS from Salmonella belonging to serogroups A, B, and D1, which are built up of tetrasaccharide-repeating units identical except for the nature of the 3,6-dideoxyhexopyranosyl group (R). : FORMULA:(SEE TEXT). The phage 28B and 36 endorhamnosidases hydrolyzed also an LPS from which the 3,6-dideoxyhexosyl substituents had previously been hydrolyzed off. However, neither of the enzymes was active on LPS preparations in which the C2-C3 bond of the L-rhamnopyranosyl ring had been opened by periodate oxidation. Glucosylation at O-6 of the D-galactopyranosyl residues in the S. typhimurium LPS was found to be incompatible with hydrolysis by both enzymes. However, in an LPS glucosylated at O-4 of the D-galactopyranosyl residues, the adjacent alpha-L-rhamnopyranosyl linkages were found to be perferentially cleaved.

Glycoside Hydrolases↗
PubMed1991

Epidemiology of Salmonella typhimurium infection in calves: persistence of salmonellae on calf units.

Salmonella typhimurium DT204C infection is the commonest cause of salmonellosis in calves. On five calf rearing farms a distinct strain, as indicated by plasmid profile analysis, was found to have persisted on the premises for periods ranging from four months to two years, the average being 14 months. The persistence of salmonellae in the environment appears to be an important factor in the epidemiology of calf salmonellosis and clearly indicates the inadequacy of many cleaning and disinfection routines.

Animal Husbandry↗
PubMed1983

Clinical salmonellosis in a guinea pig colony caused by a new Salmonella serotype, Salmonella ochiogu.

During a severe outbreak of clinical salmonellosis in an experimental guinea pig colony, a new strain of Salmonella was isolated and identified. The new serotype, with the antigenic structure 1,3,19 : Z38 : e, n, Z15, for which the name Salmonella ochiogu has been suggested, caused both enteric and systemic infection in the animal colony. During the outbreak a total of 127 animals died (26.9%). All ages of animal were affected. Treatment with oral tetracycline was successful when combined with strict hygienic measures.

Animals↗
PubMed1998

Salmonella in broiler chickens in Thailand with special reference to contamination of retail meat with Salmonella enteritidis.

The present study was performed to assess the prevalence of Salmonella in chickens in Thailand. In 1997, 22 serovars of Salmonella were isolated from 72 of 100 chicken meat samples purchased from 10 retail markets in Bangkok and 20 of 200 chicken meat samples from one slaughterhouse for export and 19 of 285 chicken feces obtained from three farms located in the east region of Thailand. The most predominant serovar was S. Enteritidis, which was isolated from 28% of the retail chicken meat, 4.5% of the chicken meat from slaughterhouse, and 6.6% of chickens feces samples examined.

Abattoirs↗
PubMed1999

Growth of Salmonella typhimurium and Salmonella enteritidis in egg yolks from highly immunized hens.

This experiment was conducted to ascertain whether the growth of Salmonella Enteritidis (SE) or Salmonella Typhimurium (ST) would be suppressed in the presence of antibodies contained in egg yolks. Specific pathogen-free chickens (102 days of age) were subcutaneously immunized with oil-adjuvanted bacterin of SE or ST, twice within a four-week interval. During 160 to 170 days of age, eggs were collected, the yolks were removed and mixed with an equal volume of physiological buffered saline, inoculated with ten colony forming units (CFU) of SE or ST, and incubated at 37 degrees C or 20 degrees C for 23 hr. The growth of organisms in each yolk solution was examined. The egg yolk derived from non-immunized hens was examined in the same manner as the controls. There was no difference in the growth titer between the antibody-positive yolk and the negative yolk. The result suggests that the antibodies in the yolk do not influence the growth of each organism, even if the hens are highly immunized.

Animals↗
PubMed1991

Detection of Salmonella serogroup D-specific antibodies in the yolks of eggs laid by hens infected with Salmonella enteritidis.

Eggs laid by hens experimentally infected with Salmonella enteritidis were assayed for the presence of Serogroup D-specific yolk antibodies. Yolk antibodies were detected with S. enteritidis and Salmonella pullorum antigens in the microantiglobulin test as early as 9 days after inoculation of hens with S. enteritidis. Yolk antibody titers reached peak levels at 3 to 5 wk postinoculation and remained at detectable levels for at least 7 wk postinoculation in eggs from both orally inoculated and horizontally contact-exposed hens. Eggs laid by hens from commercial flocks implicated in epidemiological investigations of human S. enteritidis outbreaks were also tested. Serogroup D-specific yolk antibodies were detected in 5 to 22% of eggs from hens in houses identified as infected by bacteriological culturing of internal organs of hens for S. enteritidis.

Agglutination Tests↗
PubMed1997

[Efficacy of maternal Salmonella antibodies and experimental oral infection of chicks with Salmonella enteritidis].

Distribution of maternally transmitted Salmonella antibodies and their protective effects were studied in the progeny of broiler breeder birds which had been vaccinated with live S. Typhimurium and inactivated S. Enteritidis vaccines. Vaccination resulted in a significant increase of the antibody concentration in yolk of hatching eggs and in serum and jejunum of the progeny of immunized breeder birds. Higher antibody titres for isotypes IgG and IgA were still seen on day 21 of age. Antibody production of isotypes IgA and IgM by the chickens themselves was found between 14 and 21 days of age. Two challenge models (10(2) cfu/bird on day 1 of age and a seeder bird model, respectively) were used to evaluate the efficacy of maternal antibodies against challenge with S. Enteritidis. Using both models numbers of challenge organisms were lower in the caeca of the progeny of immunized parent birds between day 7 and day 21 of age (maximum about 1.5 log10 units) compared with control chicks. The results indicate the efficacy of maternally transferred antibodies but it remains the question of their practical relevance. The effects of acquired maternal antibodies on an active immunization of the progeny of immunized breeder birds with live Salmonella vaccines are discussed.

Animals↗
PubMedDate not supplied

[Increasing number of Salmonella paratyphi B isolates from slaughtered poultry sent in to the national Salmonella reference laboratory].

In the last years the number of isolations of Salmonella enterica subspecies enterica serovar paratyphi B (S. paratyphi B) sent to the national salmonella reference laboratory of Germany has increased steadily. Most of the isolates originated from fowl or poultry products. The bacteriological, serological and biochemical properties of the isolates were investigated. Special emphasis was given to the utilization of d-tartrate which subgroups the serovar. All of them belonged to the d-tartrate positive variant, which is generally considered less virulent for humans and was formerly called S. java. The performance of various tests is compared and in addition the possibility of the spread within the production line is discussed.

Animals↗
PubMedDate not supplied

Bacteriological and serological examination and risk factor analysis of Salmonella occurrence in sow herds, including risk factors for high Salmonella seroprevalence in receiver finishing herds.

A strong association between the seroprevalence in sows and the occurrence of Salmonella Typhimurium among weaners has been shown. As shown several times for finisher herds, the risk-factors, ready mixed pelleted feed and health status also apply to sow herds. Risk factors on the sow level, for high seroprevalence in finishers have been quantified. It has been shown, that isolating Salmonella in weaners is a risk factor for high seroprevalence in finishers. Feed factors; ready-mixed pelleted feed for both sows and finishers, dry feed for sows, have been shown to have a significant effect on high seroprevalence, monitored by meat juice samples at slaughter. The etiological fraction of ready-mixed pelleted feed for sows and for finishers is of the same magnitude, indicating that intervention on the sow level could prove to contribute considerably to the effect of intervention programs.

Animal Feed↗