Serovar Strains Research Articles

A novel octavalent combined Erysipelas, Parvo and Leptospira vaccine provides (cross) protection against infection following challenge of pigs with 9 different Leptospira interrogans serovars.

BackgroundIn the present study the efficacy and cross protection of a novel ready-to-use combination vaccine, Porcilis® Ery + Parvo + Lepto, against swine leptospirosis, was investigated. The octavalent vaccine contains inactivated antigens of Erysipelothrix rhusiopathiae, Parvovirus and 6 Leptospira interrogans sensu lato strains of serogroups Canicola, Icterohaemorrhagiae, Grippotyphosa, Australis (Bratislava), Pomona and Tarassovi. In this study ninety pigs were vaccinated twice with Porcilis® Ery + Parvo + Lepto at six and ten weeks of age and ninety age and source-matched animals served as unvaccinated control. Two weeks after booster vaccination, groups of vaccinated and control pigs (ten pigs per group) were challenged with fresh virulent cultures of either of the nine different challenge strains. Compared to the vaccine strains the challenge strains were heterologous strains of the same serovar or of a different serovar within the same serogroup. The challenge strains tested were of serovar Canicola, Icterohaemorrhagiae, Copenhageni (serogroup Icterohaemorrhagiae), Grippotyphosa, Bananal/Liangguan (serogroup Grippotyphosa), Pomona, Tarassovi and Vughia (serogroup Tarassovi).ResultsAfter the different challenges most control animals became leptospiraemic for 2–7 days. The vaccinated pigs remained blood culture negative except for two animals after serovar Icterohaemorrhagiae and two animals after serovar Tarassovi challenge which became leptospiraemic for only 1 day. The incidence of Leptospiraemia (as determined by blood culture) was significantly less in vaccinates compared to the controls after all challenges. The vaccine also prevented renal infection and urinary shedding after serovar Canicola challenge. The other serovars did not induce detectable renal infection or urinary shedding.ConclusionThe present study demonstrates that the new combination vaccine Porcilis® Ery + Parvo + Lepto induces significant (cross) protection against nine different serovars within the serogroups Canicola, Icterohaemorrhagiae, Grippotyphosa, Australis (Bratislava), Pomona and Tarassovi.

Foodborne transmission of Listeria monocytogenes via ready-to-eat salad: A nationwide outbreak in Switzerland, 2013–2014

From 26 October 2013 to 23 April 2014, 32 cases of listeriosis infected with an Listeria monocytogenes strain serovar 4b, sequence type 4 and belonging to a single distinct PFGE pulsotype were registered in patients from several cantons of Switzerland. L. monocytogenes was detected in blood (75%), CSF (16%), ascites (6%) and in joint fluid (3%) samples. By the end of March 2014, a food producing company reported an L. monocytogenes contamination of ready-to-eat salads to the authorities after detecting the pathogen through its in-house routine quality control. Product and environmental samples collected during subsequent investigations yielded isolates, matching the outbreak strain, thus confirming that ready-to-eat salad from this company was most likely the outbreak source. The cause for the product contamination was related to a design-inherent hygienic problem of one specific product-feeding belt. Complementary patient interviews also identified ready-to-eat green salads bought at one retailer as the likely outbreak source.

Whole-Genome Sequence of Listeria monocytogenes Serovar 4b Strain IZSAM_Lm_hs2008, Isolated from a Human Infection in Italy.

Listeria monocytogenes is one of the most important foodborne pathogens. In this report, we present the complete and annotated genome of L. monocytogenes sequence type 06 (ST06) serovar 4b strain IZSAM_Lm_hs2008, isolated from an adult immunocompetent patient who developed the disease and died.

Contribution of the hdeB-like Gene (SEN1493) to Survival of Salmonella enterica Enteritidis NalR at pH 2

Periplasmic proteins are particularly vulnerable to denaturation upon entry into a highly acid environment. In Escherichia coli, a level of protection of these proteins is afforded by acid-inducible chaperonins encoded by hdeAB. In contrast, Salmonella enterica only harbors an hdeB-like gene and it is currently not known what function it plays in this genus. In the present study, the hdeB-like gene was deleted in Salmonella enterica Enteritidis Nal(R) and Salmonella enterica Kentucky 3795. When grown overnight in tryptic soy broth (TSB) medium buffered at pH 5.5 and then exposed to TSB pH 2 for 20 min, Enteritidis wild-type strain experienced a 0.5-log10 reduction in colony-forming units, whereas the deletion strain's surviving cells were reduced by 1.6 log10. No difference in survival was observed in the corresponding Salmonella enterica Kentucky 3795 strains treated the same way. Exposure of the strains to pH 2.5 or 3 resulted in the same log reduction regardless of the presence of the hdeB-like gene. When wild-type and deletion strains of both serovars were grown in medium buffered at pH 7 prior to exposure to the acidic pHs, no difference in survival with respect to serovar or presence/absence of the hdeB-like gene was found. Salmonella enterica Enteritidis Nal(R) carrying its own or the intragenic region upstream of the hdeB-like from Salmonella enterica Kentucky 3795 cloned in front of the gfp gene from pFPV25 showed maximum fluorescence when grown at pH 5.5, whereas the corresponding plasmid-carrying Salmonella enterica Kentucky strains did not exhibit fluorescence regardless of the pH of the growth medium. Therefore, the hdeB-like gene in Salmonella enterica Enteritidis, but not in Salmonella enterica Kentucky 3795, contributed to survival at pH 2 and its expression is responsive to the pH of the medium.

Complete Genome Sequence of Listeria monocytogenes NTSN, a Serovar 4b and Animal Source Strain.

Listeria monocytogenes is an important foodborne pathogen that causes infections in humans and animals and has a high mortality rate. The complete genome sequence of L. monocytogenes strain NTSN, a highly virulent and serovar 4b strain isolated from the brains of sheep in Jiangsu Province, China, is presented here.

Identification of putative virulence-associated genes among Haemophilus parasuis strains and the virulence difference of different serovars

This study was aimed at determining virulence-associated genes among Haemophilus parasuis (H. parasuis) strains, and supplying for the Kielstein–Rapp–Gabrielson serotyping scheme. The subtractive fragments, obtained through suppression subtractive hybridization and reverse Southern blot hybridization, were found to encode genes representative of 7 different functions. PCR was used to investigate the distribution of these fragments in H. parasuis strains isolated from different infection sites in pigs. Mice challenge was then used to analyze the correlationship between subtractive fragments, infection sites and bacterial virulence. Eight weeks old female BALB/c mice (10 mice/group) were inoculated intraperitoneally with 3.0 × 109 CFU suspension (0.5 ml/mouse) of H. parasuis strains in PBS. Results indicated that H. parasuis possessed varied virulence even among the same serovar strains. Transcription units hsdR, hsdS, gpT and ompP2, identified from the subtractive fragments, were uniformly expressed in highly virulent strains, while absent in weakly virulent strains, and demonstrated variable degrees of expression in moderately virulent strains. Moreover, H. parasuis strains, isolated from pericardium and heart blood, were all highly virulent strains, while from nasal cavity and joint were moderately or weakly virulent strains. This study indicated that fragments hsdR, hsdS, gpT and ompP2 were associated with the virulence of H. parasuis. The virulence of H. parasuis strains isolated from different infection sites was different. The current research provides a new reference for determining bacterial virulence in different H. parasuis strains.

Correlation between Ureaplasma subgroup 2 and genitourinary tract disease outcomes revealed by an expanded multilocus sequence typing (eMLST) scheme.

The multilocus sequence typing (MLST) scheme of Ureaplasma based on four housekeeping genes (ftsH, rpL22, valS, and thrS) was described in our previous study; here we introduced an expanded MLST (eMLST) scheme with improved discriminatory power, which was developed by adding two putative virulence genes (ureG and mba-np1) to the original MLST scheme. To evaluate the discriminatory power of eMLST, a total of 14 reference strains of Ureaplasma serovars and 269 clinical strains (134 isolated from symptomatic patients and 135 obtained from asymptomatic persons) were investigated. Our study confirmed that all 14 serotype strains could successfully be differentiated into 14 eMLST STs (eSTs), while some of them could not even be differentiated by the MLST, and a total of 136 eSTs were identified among the clinical isolates we investigated. In addition, phylogenetic analysis indicated that two genetically significantly distant clusters (cluster I and II) were revealed and most clinical isolates were located in cluster I. These findings were in accordance with and further support for the concept of two well-known genetic lineages (Ureaplasma parvum and Ureaplasma urealyticum) in our previous study. Interestingly, although both clusters were associated with clinical manifestation, the sub-group 2 of cluster II had pronounced and adverse effect on patients and might be a potential risk factor for clinical outcomes. In conclusion, the eMLST scheme offers investigators a highly discriminative typing tool that is capable for precise epidemiological investigations and clinical relevance of Ureaplasma.

Comparison of Widely Used Listeria monocytogenes Strains EGD, 10403S, and EGD-e Highlights Genomic Differences Underlying Variations in Pathogenicity

ABSTRACTFor nearly 3 decades, listeriologists and immunologists have used mainly three strains of the same serovar (1/2a) to analyze the virulence of the bacterial pathogen Listeria monocytogenes. The genomes of two of these strains, EGD-e and 10403S, were released in 2001 and 2008, respectively. Here we report the genome sequence of the third reference strain, EGD, and extensive genomic and phenotypic comparisons of the three strains. Strikingly, EGD-e is genetically highly distinct from EGD (29,016 single nucleotide polymorphisms [SNPs]) and 10403S (30,296 SNPs), and is more related to serovar 1/2c than 1/2a strains. We also found that while EGD and 10403S strains are genetically very close (317 SNPs), EGD has a point mutation in the transcriptional regulator PrfA (PrfA*), leading to constitutive expression of several major virulence genes. We generated an EGD-e PrfA* mutant and showed that EGD behaves like this strain in vitro, with slower growth in broth and higher invasiveness in human cells than those of EGD-e and 10403S. In contrast, bacterial counts in blood, liver, and spleen during infection in mice revealed that EGD and 10403S are less virulent than EGD-e, which is itself less virulent than EGD-e PrfA*. Thus, constitutive expression of PrfA-regulated virulence genes does not appear to provide a significant advantage to the EGD strain during infection in vivo, highlighting the fact that in vitro invasion assays are not sufficient for evaluating the pathogenic potential of L. monocytogenes strains. Together, our results pave the way for deciphering unexplained differences or discrepancies in experiments using different L. monocytogenes strains.

Epidemiology and Etiology Characteristics of Foodborne Outbreaks Caused by Vibrio parahaemolyticus During 2008–2010 in Guangdong Province, China

Vibrio parahaemolyticus infection has been considered the leading cause of bacterial illnesses mainly associated with seafood consumption in Guangdong province in China. In this study, epidemiological and etiological characteristics of 36 V. parahaemolyticus outbreaks that occurred from 2008 to 2010 in Guangdong province were analyzed; 284 strains involved were characterized by serotyping; virulence genes and 66 strains from four outbreaks therein were subtyped by pulsed-field gel electrophoresis (PFGE). Epidemiological evidence showed that 36% (13/36) of outbreaks were caused by food contamination, of which 84.6 % (11/13) were related to salted food, including viscera of pigs and cattle, meat, and vegetable salad. It was also indicated that 88.9% (32/36) of V. parahaemolyticus outbreaks appeared from June to September, 44.4% (16/36) of which occurred in canteens as well as 41.7% (15/36) in restaurants. As for the etiology, 31% (11/36) of outbreaks were caused by single serovar of strains, while 69% (25/36) were caused by multiserovars; O3:K6, O4:K8, O1:Kut, and O2:K3 were the dominant serovars. Among the 284 strains, 98.8% (254/257) of strains from patients were tdh-present and trh-absent, whereas 37.0% (10/27) from food were tdh-present. Cluster analysis of PFGE patterns demonstrated that strains in the same outbreak with identical serovar seemed to be diversified, whereas strains with various serovars could be closely related genetically. Moreover, cross-contamination between salted food and seafood was first confirmed by molecular subtyping in Guangdong, revealing that salted food might be a vital risk factor associated with V. parahaemolyticus outbreaks.

P5.081 Analytical Evaluation of the GeneXpert ® CT/NG, the First Genetic Point of Care Assay For Simultaneous Detection of Neisseria Gonorrhoeae and Chlamydia Trachomatis

IntroductionNew assays for molecular detection of Neisseria gonorrhoeae and Chlamydia trachomatis need to be evaluated for potential false positive and false negative results due to cross reaction with other species,...

P5.064 Preliminary Analytical Evaluation of Artus® CT/NG, For Simultaneous Detection of Neisseria Gonorrhoeae and Chlamydia Trachomatis

IntroductionCurrently there are a number of molecular amplification assays for detection of Neisseria gonorrhoeae and Chlamydia trachomatis. Prior to introduction of any new assay, it should be thoroughly evaluated for...

Population Structure of Salmonella enterica Serovar 4,[5],12:b:− Strains and Likely Sources of Human Infection

Salmonella enterica serovar 4,[5],12:b:- is a monophasic serovar not able to express the second-phase flagellar antigen (H2 antigen). In Germany, the serovar is occasionally isolated from poultry, reptiles, fish, food, and humans. In this study, a selection of 67 epidemiologically unrelated Salmonella enterica serovar 4,[5],12:b:- strains isolated in Germany between 2000 and 2011 from the environment, animal, food, and humans was investigated by phenotypic and genotypic methods to better understand the population structure and to identify potential sources of human infections. Strains of this monophasic serovar were highly diverse. Within the 67 strains analyzed, we identified 52 different pulsed-field gel electrophoresis XbaI profiles, 12 different multilocus sequence types (STs), and 18 different pathogenicity array types. The relatedness of strains based on the pathogenicity gene repertoire (102 markers tested) was in good agreement with grouping by MLST. S. enterica serovar 4,[5],12:b:- is distributed across multiple unrelated eBurst groups and consequently is highly polyphyletic. Two sequence types (ST88 and ST127) were linked to S. enterica serovar Paratyphi B (d-tartrate positive), two single-locus variants of ST1583 were linked to S. enterica serovar Abony, and one sequence type (ST1484) was associated with S. enterica serovar Mygdal, a recently defined, new serovar. From the characterization of clinical isolates and those of nonhuman origin, it can be concluded that the potential sources of sporadic human infections with S. enterica serovar 4,[5],12:b:- most likely are mushrooms, shellfish/fish, and poultry.

The in vitro evaluation of anti-chlamydial and cytotoxic properties of dermaseptin S4 and derivatives: peptides from amphibian skin

Chlamydia trachomatis is an obligate intracellular bacterium responsible for a number of health problems, including sexually transmitted infection in humans. Efforts were made for the search of alternative therapies. Accordingly, the present study was undertaken to perform a systemic in vitro investigation on the anti-chlamydial potential of cationic peptides from frog’s skin, namely dermaseptin S4 (S4) and its derivatives. Several strains of Chlamydia trachomatis serovar E were used to detect the antimicrobial activity of the new compounds. The infections tests and the toxic effects of the new compounds were determined using McCoy cells monolayers. Our data show that S4 exhibited a potent anti-chlamydial activity and found that these peptides blocked infection of McCoy cells and reduced the numbers of inclusion-forming units (81 %) after 48 h at low concentration (5 μg/ml). Besides, the finding revealed that increasing the number of positive charges of the peptide resulted in a reduced cytotoxicity without affecting the antimicrobial effect. Among all peptides, the derivative K4K20S4 was the more potent to inhibit C. trachomatis growth with 96 % reduction in the number of chlamydial inclusions compared with an untreated control infection and, therefore, can be considered as potential agents for therapy of Chlamydia infectious diseases.

Differences in Haemophilus parasuis adherence to and invasion of AOC-45 porcine aorta endothelial cells

BackgroundThe pathogenesis of Haemophilus parasuis depends on the bacterium’s ability to interact with endothelial cells and invade adjacent tissues. In this study, we investigated the abilities of eight H. parasuis reference strains belonging to serovars 1, 2, 4, 5, 7, 9, 10 and 13 to adhere to and invade porcine aortic endothelial cells (AOC-45 cell line).ResultsThe strains belonging to serovars 1, 2 and 5 were able to attach at high rates between 60 and 240 min of incubation, and serovars 4, 7 and 13 had moderate attachment rates; however, the strains belonging to serovars 9 and 10 had low adherence at all time points. Strong adherence was observed by scanning electron microscopy for the strains of serovars 5 and 4, which had high and moderate numbers, respectively, of H. parasuis cells attached to AOC-45 cells after 240 min of incubation. The highest invasiveness was reached at 180 min by the serovar 4 strain, followed by the serovar 5 strain at 240 min. The invasion results differed substantially depending on the strain.ConclusionThe reference strains of H. parasuis serovars 1, 2, 4 and 5 exhibited high adhesion and invasion levels to AOC-45 porcine aorta endothelial cells, and these findings could aid to better explain the pathogenesis of the disease caused by these serovars.

Fingerprints by pulsed-field gel electrophoresis of leptospires isolated from field rats and comparison with reference Leptospira serovars

Background: More than 260 leptospiral serovars by serology have been recorded. New genotypic methods have classified Leptospira into 20 species. Serovar identification is essential for epidemiological study of this disease. Objective: We identified fingerprints from reference serovar strains of Leptospira spp. and representative serovars from field rat isolates by pulsed-field gel electrophoresis (PFGE). Methods: Extract genomic DNA from 28 reference serovars of Leptospira spp. and 13 leptospiral isolates from field rats, selected as representative serovars Pyrogenes, Bataviae, Autumnalis, and Australis, using PFGE following NotI restriction digest. Results: PFGE with NotI restriction enzyme successfully differentiated 28 reference serovars into 27 fingerprint patterns, with the exception of serovars Copenhageni and Icterohaemorrhagiae. The discriminatory power of these reference strains was 0.99. Isolates that yielded patterns identical to their corresponding serovars were serovars Pyrogenes strain Salinem and Bataviae. Patterns for isolates of serovars Autumnalis and Australis were different from reference serovar Autumnalis strain Akiyami A and serovar Australis strain Ballico used in this study. Conclusion: PFGE can be useful for identifying serovars of leptospiral isolates from reservoirs and for identifying new serovars of Leptospira for epidemiological study. Keywords: Leptospira, leptospiral serovar, molecular typing, PFGE, pulsed-field gel electrophoresis

Identification of Salmonella enterica species- and subgroup-specific genomic regions using Panseq 2.0

The pan-genome of a taxonomic group consists of evolutionarily conserved core genes shared by all members and accessory genes that are present only in some members of the group. Group- and subgroup-specific core genes are thought to contribute to shared phenotypes such as virulence and niche specificity. In this study we analyzed 39 Salmonella enterica genomes (16 closed, 23 draft), a species that contains two human-specific serovars that cause typhoid fever, as well as a large number of zoonotic serovars that cause gastroenteritis in humans. Panseq 2.0 was used to define the pan-genome by adjusting the threshold at which group-specific “core” loci are defined. We found the pan-genome to be 9.03Mbp in size, and that the core genome size decreased, while the number of SNPs/100bp increased, as the number of strains used to define the core genome increased, suggesting substantial divergence among S. enterica subgroups. Subgroup-specific “core” genes, in contrast, had fewer SNPs/100bp, likely reflecting their more recent acquisition. Phylogenetic trees were created from the concatenated and aligned pan-genome, the core genome, and multi-locus-sequence typing (MLST) loci. Branch support increased among the trees, and strains of the same serovar grouped closer together as the number of loci used to create the tree increased. Further, high levels of discrimination were achieved even amongst the most closely related strains of S. enterica Typhi, suggesting that the data generated by Panseq may also be of value in short-term epidemiological studies. Panseq provides an easy and fast way of performing pan-genomic analyses, which can include the identification of group-dominant as well as group-specific loci and is available as a web-server and a standalone version at http://lfz.corefacility.ca/panseq/.

Genomic characterization of Haemophilus parasuis SH0165, a highly virulent strain of serovar 5 prevalent in China.

Haemophilus parasuis can be either a commensal bacterium of the porcine respiratory tract or an opportunistic pathogen causing Glässer's disease, a severe systemic disease that has led to significant economical losses in the pig industry worldwide. We determined the complete genomic sequence of H. parasuis SH0165, a highly virulent strain of serovar 5, which was isolated from a hog pen in North China. The single circular chromosome was 2,269,156 base pairs in length and contained 2,031 protein-coding genes. Together with the full spectrum of genes detected by the analysis of metabolic pathways, we confirmed that H. parasuis generates ATP via both fermentation and respiration, and possesses an intact TCA cycle for anabolism. In addition to possessing the complete pathway essential for the biosynthesis of heme, this pathogen was also found to be well-equipped with different iron acquisition systems, such as the TonB system and ABC-type transport complexes, to overcome iron limitation during infection and persistence. We identified a number of genes encoding potential virulence factors, such as type IV fimbriae and surface polysaccharides. Analysis of the genome confirmed that H. parasuis is naturally competent, as genes related to DNA uptake are present. A nine-mer DNA uptake signal sequence (ACAAGCGGT), identical to that found in Actinobacillus pleuropneumoniae and Mannheimia haemolytica, followed by similar downstream motifs, was identified in the SH0165 genome. Genomic and phylogenetic comparisons with other Pasteurellaceae species further indicated that H. parasuis was closely related to another swine pathogenic bacteria A. pleuropneumoniae. The comprehensive genetic analysis presented here provides a foundation for future research on the metabolism, natural competence and virulence of H. parasuis.

Survival of Salmonella Newport in oysters

Salmonella enterica is the leading cause of laboratory-confirmed foodborne illness in the United States and raw shellfish consumption is a commonly implicated source of gastrointestinal pathogens. A 2005 epidemiological study done in our laboratory by Brands et al., showed that oysters in the United States are contaminated with Salmonella, and in particular, a specific strain of the Newport serovar. This work sought to further investigate the host-microbe interactions between Salmonella Newport and oysters. A procedure was developed to reliably and repeatedly expose oysters to enteric bacteria and quantify the subsequent levels of bacterial survival. The results show that 10 days after an exposure to Salmonella Newport, an average concentration of 3.7 × 10(3)CFU/g remains within the oyster meat, and even after 60 days there still can be more than 10(2)CFU/g remaining. However, the strain of Newport that predominated in the market survey done by Brands et al. does not survive within oysters or the estuarine environment better than any other strains of Salmonella we tested. Using this same methodology, we compared Salmonella Newport's ability to survive within oysters to a non-pathogenic strain of E. coli and found that after 10 days the concentration of Salmonella was 200-times greater than that of E. coli. We also compared those same strains of Salmonella and E. coli in a depuration process to determine if a constant 120 L/h flux of clean seawater could significantly reduce the concentration of bacteria within oysters and found that after 3 days the oysters retained over 10(4)CFU/g of Salmonella while the oysters exposed to the non-pathogenic strain of E. coli contained 100-times less bacteria. Overall, the results of this study demonstrate that any of the clinically relevant serovars of Salmonella can survive within oysters for significant periods of time after just one exposure event. Based on the drastic differences in survivability between Salmonella and a non-pathogenic relative, the results of this study also suggest that unidentified virulence factors may play a role in Salmonella's interactions with oysters.

Characterization of Haemophilus parasuis isolated from Brazilian swine through serotyping, AFLP and PFGE

Haemophilus parasuis infection in pigs is characterized by fibrinous polyserositis, arthritis and meningitis. Despite the fact that traditional diagnosis is based on herd history, clinical signs, bacterial isolation and serotyping, molecular-based methods are alternatives for species-specific tests and epidemiological studies. The aim of this study was to characterize H. parasuis field strains from different states of Brazil, employing serotyping and genotyping methods. Serotyping revealed that serovar 4 was the most prevalent (26.1%), followed by serovars 5 (17.4%), 14 (8.7%), 13 (4.4%) and 2 (4.4%), whereas 39% of the strains were considered as untypeable. AFLP with a single enzyme and PFGE were able to type all isolates tested, generating 34 and 20 different profiles, respectively, including untypeable strains. Besides the slightly higher discrimination index presented by AFLP, PFGE with Not I restriction enzyme showed a better correlation with epidemiological data, grouping strains of the same serovar, animal or farm origin. The results indicated AFLP and PFGE as valuable tools for typing H. parasuis isolates collected in Brazil.

Exploitation of Intestinal Colonization‐Inhibition Between Salmonella Organisms for Live Vaccines in Poultry – Potential and Limitations

Immunization represents one of the most important methods to increase the resistance of chickens against Salmonella infection. In addition to the development of an adaptive immune response, oral administration of live Salmonella strains to day-old chicks provides protection against infection within hours by intestinal colonization-inhibition. For the exploitation of this phenomenon, practical information on colonization-inhibition between Salmonella organisms is needed. Colonization-inhibition capacity between Salmonella strains from serogroups B, C1, C2, D and G was assessed in chickens. The most profound level of intestinal colonization-inhibition occurred between isogenic strains. Inhibition between strains of the same serovar was greater than that between strains of different serovars. The degree of inhibition between different serovars was not sufficiently high to identify a single strain which might inhibit a wide range of other Salmonella organisms. However, as Salmonella Enteritidis is the dominant serovar in poultry in many countries and because of the profound colonization-inhibition within this serovar there is a considerable potential to exploit this phenomenon in the development of novel live S. Enteritidis vaccines. Treatment of young chicks with mixtures of different Salmonella serovars resulted not only in a very strong growth inhibition of the isogenic strains but also in a substantial inhibition of heterologous serovars. The potential of mixtures of heterologous Salmonella strains as a 'Salmonella Inhibition Culture' and as a 'live Salmonella vaccine' should be further explored.