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PubMed2005

Evaluation of a French ELISA for the detection of Salmonella Enteritidis and Salmonella Typhimurium in flocks of laying and breeding hens.

In France, the regular and compulsory detection of Salmonella Enteritidis (SE) and Salmonella Typhimurium (ST) in flocks of breeding and laying hens is based on bacteriological examination of environmental swabs and faeces samples. The aim of this study was to compare this bacteriological examination with a serological method (ELISA) developed in our laboratory. This ELISA was first evaluated by use of artificially infected hens. During these experimental infection studies, several groups of hens were inoculated with SE, ST, different vaccines and different Salmonella serovars to calculate the experimental parameters of our ELISA. Then, in a field study, 43 flocks were followed monthly using two bacteriological samples (environmental swab and pool of faeces) and 20 serological samples (sera or yolks). Twenty-seven flocks without SE or ST gave a negative serological response throughout their surveillance. Among the 10 various serovars different from SE and ST isolated in this study, S. Heidelberg, S. Agona and S. Hadar gave seropositive results in seven flocks. Consequently, this ELISA was not specific of SE and ST as it detected serovars sharing or not common antigens with SE and ST. Seropositive results were also obtained each month for two flocks where no Salmonella could be isolated. Finally, in seven flocks found infected with SE or ST, the positive ELISA results appeared later than the bacteriological detection. Therefore, for the detection of chicken flocks recently infected with SE or ST, bacteriological examination currently used in France seems to be more appropriate than this ELISA.

Animals↗

PubMed2000

Orally administered attenuated Salmonella enteritidis reduces chicken cecal carriage of virulent Salmonella challenge organisms.

Chickens were immunized orally with 10(9)cfu of the temperature-sensitive (T(s)) mutant E/1/3 of Salmonella enteritidis at 1, 2, 3 and 7 days of age. The animals were challenged with wild-type strains of Salmonella of different serotypes 7 or 14 days following immunization. Chickens receiving multiple oral doses of the vaccine strain showed no signs of disease. Immunized animals shed the vaccine strain for at least 2 weeks after the last inoculation; on the other hand, colonization by the attenuated mutant of internal organs such as spleen and liver was limited. Early exposure of the immunized animals to the virulent bacteria resulted in a reduced cecal colonization by the pathogen. Visceral invasion by the wild-type strain of S. enteritidis or S. gallinarum was drastically diminished in birds challenged 14 days after immunization. Significant differences in the number of these Salmonella were found in the cecal contents, spleen and liver of immunized birds compared with the control animals. In addition, cecal colonization by the virulent strain was reduced in birds challenged with S. typhimurium. These results demonstrate that immunization of newly hatched chickens with live attenuated T(s) mutant E/1/3 of S. enteritidis is safe and reduces Salmonella shedding.

Administration, Oral↗

PubMed2006

Impact of heat stress on the fecal shedding patterns of Salmonella enterica Typhimurium DT104 and Salmonella enterica infantis by 5-week-old male broilers.

The objective of this study was to determine if there is an impact of heat stress of broiler chickens on number and survival of two types of Salmonella shed in the chicken's feces after an oral challenge. The data from this study indicate that heat stress did not result in higher levels or longer survival of Salmonella spp. shed in feces. It is possible that the duration or intensity of the heat stress employed was not sufficient or that heat stress does not alter the number or survivability for these particular strains of Salmonella spp. Feces stored at room temperature after collection, resulted in the numbers of both strains of Salmonella increasing by one to three logs in the first week. This finding indicates that there could be an increase in environmental contamination under certain conditions.

Animals↗

PubMed2001

Persistence of a Salmonella enterica serovar typhimurium DT12 clone in a piggery and in agricultural soil amended with Salmonella-contaminated slurry.

Prevalence of Salmonella enterica on a Danish pig farm presenting recurrent infections was investigated. A comparison of the pulsed-field gel electrophoresis patterns of fecal isolates from piggeries, waste slurry, and agricultural soil amended with Salmonella-contaminated animal waste (slurry) and subclinical isolates from the same farm (collected in 1996 and later) showed identical patterns, indicating long-term persistence of the Salmonella enterica serovar Typhimurium DT12 clone in the herd environment. Furthermore, when Salmonella-contaminated slurry was disposed of on the agricultural soil (a common waste disposal practice), the pathogen was isolated up to 14 days after the spread, indicating potentially high risks of transmission of the pathogen in the environment, animals, and humans.

Animal Husbandry↗

PubMed2005

Genomic comparisons of Salmonella enterica serovar Dublin, Agona, and Typhimurium strains recently isolated from milk filters and bovine samples from Ireland, using a Salmonella microarray.

Salmonella-induced enterocolitis is the leading food-borne illness with a lethal outcome and causes millions of cases of gastroenteritis each year. We examined genomic variation among 12 environmental, veterinary, and clinical Salmonella enterica serovar Dublin, Agona, and Typhimurium strains isolated in Ireland between 2000 and 2003, as well as two clinical isolates from Canada and four archival isolates, which belonged to serovars Dublin and Agona. Using DNA-DNA hybridization to a microarray consisting of most of the predicted protein-encoding sequences of the S. enterica serovar Typhimurium LT2 genome, we identified a number of genomic regions that were absent in one or more serovars. The 34 genomic regions encoded proteins involved in sugar metabolism, transport, fimbrial and phage biogenesis, and transcriptional regulation, as well as inner and outer membrane-associated proteins. Two of the four prophages identified in strain LT2, prophages Gifsy-1 and Gifsy-2, were present in all six serovar Typhimurium strains examined. Prophage Fels-1 was absent from all 18 isolates examined, and Fels-2 was completely absent from the serovar Typhimurium isolates and the Salmonella Reference Collection B serovar Dublin strain Du2. All five Salmonella pathogenicity islands were present in all isolates. Plasmid pSLT was absent from all serovar Agona isolates, and only homologues of the spv genes were present in eight of the nine serovar Dublin strains. Only limited intraserovar diversity was found among the nine serovar Dublin, three serovar Agona, and six serovar Typhimurium isolates examined even though these isolates had extensive geographic, temporal, and source differences.

Animals↗

PubMed2003

Genomic comparison of Salmonella enterica serovars and Salmonella bongori by use of an S. enterica serovar typhimurium DNA microarray.

The genus Salmonella consists of over 2,200 serovars that differ in their host range and ability to cause disease despite their close genetic relatedness. The genetic factors that influence each serovar's level of host adaptation, how they evolved or were acquired, their influence on the evolution of each serovar, and the phylogenic relationships between the serovars are of great interest as they provide insight into the mechanisms behind these differences in host range and disease progression. We have used an Salmonella enterica serovar Typhimurium spotted DNA microarray to perform genomic hybridizations of various serovars and strains of both S. enterica (subspecies I and IIIa) and Salmonella bongori to gain insight into the genetic organization of the serovars. Our results are generally consistent with previously published DNA association and multilocus enzyme electrophoresis data. Our findings also reveal novel information. We observe a more distant relationship of serovar Arizona (subspecies IIIa) from the subspecies I serovars than previously measured. We also observe variability in the Arizona SPI-2 pathogenicity island, indicating that it has evolved in a manner distinct from the other serovars. In addition, we identify shared genetic features of S. enterica serovars Typhi, Paratyphi A, and Sendai that parallel their unique ability to cause enteric fever in humans. Therefore, whereas the taxonomic organization of Salmonella into serogroups provides a good first approximation of genetic relatedness, we show that it does not account for genomic changes that contribute to a serovar's degree of host adaptation.

Bacterial Proteins↗

PubMed2006

Effects of different molting procedures on incidence of Salmonella infection in flocks of naturally contaminated laying hens in a commercial egg-producing farm by detection of yolk antibodies to Salmonella in eggs.

Indirect enzyme-linked immunosorbent assays (ELISAs) have been applied to detect immunoglobulin Y antibodies to different serotypes of Salmonella in the yolks of chicken eggs with heat-extracted antigens of Salmonella enterica serotypes Agona (SA), Cerro (SC), Enteritidis (SE), Montevideo (SM), and Putten (SP). The egg yolk samples examined were classified as positive if their ELISA absorbance values exceeded the value for eggs from specific-pathogen-free flocks by more than two standard deviations. Of 30 egg yolk samples from three flocks vaccinated with a killed SE vaccine, 29 were antibody positive by the ELISA assay for the SE antigen. Four to 29 of the 29 yolk samples showed positive results for the other serovars, although the absorbance values for SE were higher than those obtained for the other serotypes in each of the yolk samples. All 30 yolks from three flocks that were not administered any SE vaccines were found to be antibody negative for SE, and two samples were determined to be positive for SC. Thirty-nine or 40 eggs were obtained from each of four layer flocks in a commercial egg production farm where the laying houses were naturally contaminated with SA, SC, SM, SP, Salmonella serovar Infantis (SI), and untypeable strains. The ELISA absorbance values for SM in the egg yolks obtained from the two flocks molted through feed withdrawal when the birds restarted laying were significantly (P < 0.05) higher than those observed in the yolks obtained before the molt. In egg yolks from the two other flocks that were molted through a wheat bran diet, there was no significant difference between the absorbance values before and after the molt. The observations in the present study provide further evidence to suggest that a molt initiated through the administration of a wheat bran diet can reduce the risk for Salmonella problems in a commercial egg-producing setting.

Animals↗

PubMed1977

[Introduction of an additional H-agglutinating test antigen for the differentiation of specific positive reactions in pullorum disease-fowl typhoid in poultry experimentally infected with Salmonella gallinarum-pullorum and Salmonella enteritidis].

Explored were the possibilities to differentiate the specific positive reactions in case of pullorum disease in experimentally infected (Salmonella gallinarum-pullorum and Salmonella enteritidis) birds with the use of a typhoid test antigen and two H test antigens, obtained by the routinely employed method with formalin from Salmonella enteritidis and Salmonella adeoyo. It was demonstrated that the H-agglutinating antibodies persisted in the peripheral blood of the experimentally infected birds for at least 30 days (as long as the test lasted). The H test antigen of S. enteritidis proved unsuitable to detect the H-agglutinating antibodies only as it retained to a large extent the O antigens. By means of the S. adeoyo H test antigen only the H-agglutinating antibodies in the peripheral blood of S. enteritidis-infected birds could be quickly and precisely demonstrated. A combined serologic examination of serum with the thyphoid test antigen and the S. adeoyo test antigen could be carried out to differentiate the specific positive reactions for establishing pullorum disease from the concomitant agglutination reactions observed in cases of experimental salmonellosis in birds caused by S. enteritidis.

Agglutination Tests↗

PubMedDate not supplied

Unusual presentation of enteric fever: three cases of splenic and liver abscesses due to Salmonella typhi and Salmonella paratyphi A.

Enteric fever is a multisystem disorder caused mainly by Salmonella typhi and Salmonella paratyphi A. It continues to be a major public health problem, especially in developing countries. Unusual presentations of Salmonellosis are rare. We report 3 such cases of young adult males, one of splenic abscess due to Salmonella typhi and one each of liver abscess due to Salmonella typhi and Salmonella paratyphi A. A brief review of the literature pertaining to the cases is also given.

Abscess↗

PubMed2002

Processing of viable Salmonella typhimurium for presentation of a CD4 T cell epitope from the Salmonella invasion protein C (SipC).

We have identified Salmonella invasion protein C (SipC) as a target antigen for CD4 T cell recognition in mice infected with Salmonella typhimurium. SipC is a product of the type III secretion system encoded by S. typhimurium pathogenicity island 1. A SipC-specific T cell response was induced by infection with either the C5 wild type or attenuated SL3261 vaccine strain of S. typhimurium. We localized the response of T cell lines from infected mice to an epitope near the carboxyl terminus of SipC (SipC(381-394)) and studied the way it was processed from viable S. typhimurium. We demonstrated that CD4 T cell recognition of this epitope required actin-dependent uptake of S. typhimurium. Presentation also occurred when transport of newly synthesized MHC class II from the endoplasmic reticulum was disrupted and when the pH of intracellular compartments was raised, suggesting presentation by mature MHC class II recycled from the macrophage surface into neutral intracellular compartments. Salmonellae are known to colonize macrophages by localizing to compartments that do not make contact with the bactericidal environment of late endosomes or lysosomes, and thus might avoid lysosomal antigen processing. However, we demonstrate that a CD4 T cell response to S. typhimurium-secreted proteins may be induced by an alternative pathway capable of antigen presentation in conditions similar to those in the compartments where Salmonella localize.

Animals↗

PubMed2004

Cellular microbiology of intracellular Salmonella enterica: functions of the type III secretion system encoded by Salmonella pathogenicity island 2.

The facultative intracellular pathogen Salmonella enterica resides in a special membrane compartment of the host cell and modifies its host to achieve intracellular survival and proliferation. The type III secretion system encoded by Salmonella pathogenicity island 2 (SPI2) has a central role in the interference of intracellular Salmonella with host cell functions. SPI2 function affects antimicrobial defense mechanisms of the host, intracellular transport processes, integrity and function of the cytoskeleton and host cell death. These modifications are mediated by translocation of a large number of effector proteins by the SPI2 system. In this review, we summarize recent work on the cellular phenotypes related to SPI2 function and contribution of SPI2 effector proteins to these manipulations. These studies reveal a complex set of pathogenic interferences between intracellular Salmonella and its host cells.

Animals↗

PubMed1992

Characterisation of monoclonal antibodies against a fimbrial structure of Salmonella enteritidis and certain other serogroup D salmonellae and their application as serotyping reagents.

A panel of 13 monoclonal antibodies from different hybridomas was produced against a novel salmonella fimbrial antigen expressed predominantly by Salmonella enteritidis strains. The specificity of the monoclonal antibodies to this antigen (SEF14) was confirmed by enzyme-linked immunosorbent assay (ELISA) using purified SEF14, immune electron microscopy and, with 11 monoclonal antibodies, the identification of a repeating protein subunit (14,300kDa) on the antigen. Blocking-ELISA with the monoclonal antibodies identified epitopes in at least three, non-overlapping clusters which appeared evenly distributed on SEF14 in immune electron microscopy. The use of the monoclonal antibodies in direct-binding ELISA on a range of salmonella serotypes suggested that the epitopes on SEF14 are highly conserved and were expressed by all the S enteritidis strains examined; some strains of S dublin and the only strain of S moscow available were the only other serotypes that expressed SEF14. A latex agglutination reagent based on a monoclonal antibody was developed and used to test for SEF14 on 280 strains (representing 120 serotypes in 24 serogroups of salmonellae) that had been grown on Sensitest agar for 18 hours at 37 degrees C. All S enteritidis strains (64) and most S dublin strains (28 of 33) produced SEF14 as did the two strains representing S blegdam and S moscow. SEF14 was not detected in any other strains of serotypes from serogroup D or from any other serogroup examined.

Antibodies, Bacterial↗

PubMed1992

Characterization of the Salmonella typhimurium phoE gene and development of Salmonella-specific DNA probes.

In Escherichia coli K-12, the phoE gene, encoding a phosphate-limitation-inducible outer membrane pore protein (PhoE), is closely linked to the genes proA and proB. When the corresponding fragment of the Salmonella typhimurium chromosome was transferred to E. coli K-12 using an RP4::miniMu plasmid, pULB113, no production of S. typhimurium PhoE could be detected. Nevertheless, DNA hybridization studies revealed that the corresponding plasmid did contain S. typhimurium phoE. Production of S. typhimurium PhoE in E. coli was detected only after subcloning the gene in a multicopy vector. Nucleotide (nt) sequence analysis showed extensive homology of S. typhimurium phoE to the E. coli gene and suggested possible explanations for the low expression of S. typhimurium phoE in E. coli. In addition, the sequence information was used to develop Salmonella-specific DNA probes. Two oligodeoxyribonucleotides were synthesized based on nt sequences encoding the fifth and eighth cell-surface-exposed regions of PhoE. When used in polymerase chain reactions, these probes turned out to be specific, i.e., no crossreactions occurred with the non-Salmonella strains, whereas 132 out of 133 tested Salmonella strains were recognized.

Amino Acid Sequence↗

PubMed1990

Virulence plasmids of Salmonella typhimurium and other salmonellae.

Related high molecular weight plasmids of several serotypes and species of Salmonella have been associated with virulence in a variety of animal models of infection. The primary virulence plasmid phenotype is in the ability of salmonellae to spread beyond the initial site of infection, the intestines. The mechanism of this plasmid-mediated invasive infection has not been identified, but may be a complex interaction in the host-pathogen relationship. A common region of the salmonella plasmids has been associated with virulence, and specific virulence genes and their products are now being identified; however, much is yet to be accomplished in this field. The combined analysis of pathogenesis and genetics associated with the salmonella virulence plasmids may identify new systems of bacterial virulence and the genetic basis for this virulence.

Animals↗

PubMed1990

Expression of host resistance to Salmonella typhi and Salmonella typhimurium: bacterial survival within macrophages of murine and human origin.

Cell-association of various strains of Salmonella typhi and Salmonella typhimurium with different populations of macrophages was studied. Macrophages were infected, exposed to gentamicin, washed, and counts of viable bacteria protected from gentamicin killing were made. J774A.1 cells, a continuous macrophage-like cell line, were the most permissive, all strains tested achieving similar high recoveries. Virulent S. typhimurium 779C-Sms, but not avirulent S. typhimurium 779C-SmD, survived well in mouse peritoneal macrophages and human monocyte-derived macrophages. Virulent S. typhi Ty2 were killed by mouse peritoneal macrophages, but were able to survive within human monocyte-derived macrophages. Viable counts of clinical isolates of S. typhi within the human monocyte-derived phagocytes were lower as compared with those of S. typhi Ty2. Phagocytosis of opsonized and non-opsonized virulent S. typhi Ty2 and S. typhimurium 779C-SmS by mouse peritoneal macrophages failed to trigger their respiratory burst as assessed by the intracellular reduction of nitroblue tetrazolium dye (NBT). These experiments support the view that the intracellular survival of Salmonella is in part host dependent and specific in nature. They also suggest that virulence influences the survival and intracellular multiplication of Salmonella within macrophages, and that their ultimate fate within macrophages may not be related to oxygen-dependent mechanisms.

Animals↗

PubMed1992

Role of T cells, TNF alpha and IFN gamma in recall of immunity to oral challenge with virulent salmonellae in mice vaccinated with live attenuated aro- Salmonella vaccines.

The SL3261 Salmonella typhimurium aroA live vaccine strain confers solid protection against oral challenge with virulent salmonellae, immunity persisting long after the vaccine has been cleared from the tissues. BALB/c mice immunized with SL3261 and later subjected to in vivo depletion of both CD4+ and CD8+ T cells had impaired recall of immunity to oral challenge with the virulent S. typhimurium C5, with increased mortality and higher bacterial loads in the reticuloendothelial system (RES). Selective depletion of CD4+ cells alone significantly impaired resistance both 8 and 14 weeks after vaccination as determined by estimation of bacterial numbers in organ homogenates. Depletion of CD8+ cells alone had less effect on immunity when performed at 8 weeks than at 14 weeks after immunization. Administration of anti-IFN gamma or anti-TNF alpha antibodies also impaired recall of immunity, exacerbating a secondary infection in vaccinated mice. Challenge of T cell-depleted immune mice with virulent salmonellae caused hepatosplenomegaly with minute grossly visible focal lesions, and a marked increase in the number and severity of necrotic foci in spleen, liver and lymph nodes. A widespread mononuclear cell infiltrate was present. The histopathology in anti-IFN gamma-treated mice was qualitatively similar to that seen in T-cell depleted mice. In contrast, in the anti-TNF alpha-treated mice splenomegaly was much less than in T cell-depleted mice. Granulomas were absent, no mononuclear infiltration was observed and there was severe necrosis; the lesions appeared similar to or worse than those seen in naïve mice. Surprisingly, IFN gamma was detectable in sera of both controls and T cell-depleted mice on day 8 of the secondary infection, as well as in sera of anti-TNF alpha-treated mice on day 6 of infection. The results indicate that T cells, IFN gamma and TNF alpha are all important in the specific recall of immunity to virulent salmonellae conferred by immunization with live vaccines, with the effect of T cell and IFN gamma depletion (marked macrophage infiltration) being qualitatively very different from that of TNF alpha neutralization (no mononuclear infiltrate or granuloma formation).

Animals↗

PubMed1981

Observations on the occurrence of Salmonella cholerae-suis and other salmonellas in two herds of feeder pigs.

Modified brilliant green agar (BGA), Muller-Kauffmann tetrathionate, Rappaport's and selenite F broths were compared for their efficiency in isolating salmonellas from pigs and their excreta. It was concluded that BGA and Rappaport's broth were the media of choice. Where searches were made for Salmonella cholerae-suis alone, the use of a trehalose McConkey agar provided a rapid method of differentiating S. cholerae-suis, which does not ferment trehalose, from the majority of other salmonellas, which do ferment trehalose. Casualties were collected from two farms where infection with S. cholerae-suis was endemic. The isolation rates of S. cholerae-suis from different carcase sites were compared in order to determine the relative importance of the salivary, upper respiratory and faecal routes of excretion. S. cholerae-suis was isolated from numerous carcase sites in carriers including the salivary glands, tonsils, trachea and lungs. However, isolations from the nasal passages, mouth, pharynx and gastro-intestinal tract of carriers were either infrequent or absent. When, in a further study, S. cholerae-suis was isolated from only 3/414 faeces, 1/170 nasal swabs and not at all from 170 oral swabs taken from live pigs, it was concluded that there must be more significant modes of transmission than from the salivary glands, upper respiratory or gastro-intestinal tracts. Cannibalism was considered to be a possibility. In contrast to S. cholerae-suis, other salmonellas were frequently isolated from randomly collected faeces and from the gastro-intestinal tract as well as other sites in casualties.

Animals↗

PubMed2006

Experimental airborne transmission of Salmonella Agona and Salmonella Typhimurium in weaned pigs.

This study tested the hypothesis of airborne transmission of Salmonella Agona (Trial I) and Salmonella Typhimurium (Trial II) in weaned pigs. The trials were performed using stainless-steel/glass isolation cabinets connected by air ducts to permit an unidirectional airflow from cabinet 1 (two control pigs) to cabinet 3 (two sentinel pigs), passing through cabinet 2 (two inoculated pigs). Air samples, pooled faecal samples from the floor and rectal swabs were collected daily and assessed by culture and PCR. A fumigation chamber and rubber gloves coupled to the cabinets allowed sampling without opening the system. Trials I and II lasted 15 and 19 days respectively. The recovery of S. Agona and S. Typhimurium and detection of seroconversion in sentinel pigs indicate that airborne Salmonella transmission in weaned pigs over short distances is possible. Further studies on the role of aerosols in the epidemiology of Salmonella in intensive pig production should be performed.

Aerosols↗