Detection of Salmonella-containing food using the 1-2 test

133 samples of food were investigated for comparison of a recent commercially available 1-2 test (Biocontrol) with a cultural standard method (non-selective pre-enrichment in buffered peptone water, selective enrichment in Rappaport-Vassiliadis-medium and selenite brilliant-green mannitol enrichment broth, inoculation on two selective agars) for presence of Salmonella. The 1-2 test showed in positive results an accuracy ("sensitivity") of 94.7% and in negative results an assurance ("specificity") of 97.9% and is therefore considered suitable for detection of Salmonella contaminated food.     

Prevalence of the Salmonella plasmid virulence gene "spvD" in Salmonella strains from animals]

Strains of Salmonella isolated from animals in Germany (n = 878) were analysed for the presence of the spvD gene ("Salmonella plasmid virulence gene D") by DNA-DNA hybridization. The spvD gene was only detected in strains of serovars Typhimurium (93.3%), Enteritidis (97.1%), and Dublin (100%) as well as in two rough strains of Salmonella enterica. Salmonella isolates from mammals carried the gene more frequently (cattle 94.0%, horses 92.6%, pigs 73.7%) than those from birds (33.3%) or reptiles (4.5%). Due to its high prevalence in epidemiologically relevant salmonellae, the virulence factor spvD may represent a sensitive and specific target in various serovars for diagnostic or immunization strategies.     

Culling decisions of dairy farmers during a 3-year Salmonella control study

Salmonella enterica subsp. enterica-serotypes lead to periodically increased morbidity and mortality in cattle herds. The bacteria can also lead to serious infections in humans. Consequently, Denmark has started a surveillance and control programme in 2002. The programme focuses on Salmonella Dublin which is the most prevalent and most persistent serotype in the Danish cattle population. A field study in 10 dairy herds with persistent Salmonella infections was carried out over three years to gain experience with control procedures including risk assessment, targeted control actions and test-and-cull procedures. From autumn 2003 until end of 2006 quarterly milk quality control samples from all lactating cows and biannual blood samples from all young stock above the age of three months were tested using an indirect antibody ELISA. The most recent and previous test results were used to categorise all animals into risk groups. These risk groups and all individual ELISA-results were communicated to the farmers as colour-coded lists four to six times per year. Farmers were advised to manage the risk of Salmonella transmission from cattle with repeatedly high ELISA results (flagged as "red") or cows with at least one recent moderately high ELISA result (flagged as "yellow") on the lists. Risk management included, e.g. culling or separation of the cows at calving. We analysed culling decisions using two models. For heifers a hierarchical multivariable logistic model with herd as random effect evaluated if animals with red and yellow flags had higher probability of being slaughtered or sold before first calving than animals without any risk flags. For adult cows a semi-parametric proportional hazard survival model was used to test the effect of number of red and yellow flags on hazards of culling at different time points and interactions with prevalence in the herd while accounting for parity, stage of lactation, milk yield, somatic cell count and the hierarchical structure of the data with animals clustered at herd level. This study illustrates how investigation of culling decisions made by herd managers when they have access to test-status of individual animals and overall apparent prevalence during control of an infection can lead to useful new knowledge. Overall herd managers were more likely to cull cattle with increasing number of yellow and red flags than animals with no flags. However, cattle were more likely to be culled with yellow and red flags during times with low or medium high within-herd seroprevalence than at times with high seroprevalence. These results are valuable knowledge for modelling and planning of control strategies and for making recommendations to farmers about control options.     

Enrichment for isolating Salmonella Choleraesuis and other Salmonella spp. from pigs

The growth of Salmonella Choleraesuis was examined in Rappaport Vassiliadis broth (RV) and Hajna-tetrathionate broth (HTT) at 37 and 42 degrees C. As the enrichment in RV at 37 degrees C was satisfactory for isolating S. Choleraesuis, we used this enrichment for isolation from the samples collected from 15 asymptomatic pigs reared on a S. Choleraesuis contaminated farm. S. Choleraesuis was frequently isolated from six pigs (40.0%) under field conditions. The isolation of other Salmonella serovars than S. Choleraesuis was attempted by using both RV enrichment at 37 degrees C and HTT enrichment at 42 degrees C. Salmonella organisms were isolated from 156 (44.8%) of 348 fecal samples and more frequently with HTT at 42 degrees C (43.4%) than with RV at 37 degrees C (20.9%). If other serovars in addition to S. Choleraesuis are to be surveyed, HTT enrichment should be used in combination with RV enrichment.     

Transmission of Salmonella in dairy herds quantified in the endemic situation

Salmonella is a cause of concern in the cattle industry, because it is a zoonosis causing severe invasive infections in humans and because it causes economic and welfare losses in infected herds. In general, cattle in the Netherlands are infected with two types; Salmonella Dublin and Salmonella Typhimurium. Both types cause clinical signs but S. Dublin outbreaks are more prevalent and clinical signs are more severe than S. Typhimurium outbreaks. Our knowledge of the transmission of Salmonella within herds is still limited, while this is an essential component for modelling the success of intervention strategies to control Salmonella. The aim of our study was to estimate the basic reproduction ratio (R(0)), the number of secondary cases produced from each primary case in a totally susceptible population, for S. Dublin and S. Typhimurium in dairy herds. Serological data were obtained from eight farms with a clinical outbreak of Salmonella, two with an outbreak of S. Dublin and 6 of S. Typhimurium. R(0) was estimated from the serological data of the herds that were in an endemic state of the infection. R(0) across herds was estimated to be 2.5 (95% CI 1.7-9.8) and 1.3 (95% CI 1.1-1.7) for S. Dublin and S. Typhimurium, respectively. The between herd variation was significant and fairly large. The results of the sensitivity analysis showed that the R(0) estimate was not sensitive for changes in the latent, infectious or seropositive periods. The R(0) estimates indicated that the infection would not spread very extensively in susceptible populations under management systems similar to the ones in the study herds.     

Use of a Salmonella typhimurium-derived virulence probe in the detection of Salmonella sp. and in the characterization of S. cholerae-suis virulence plasmids

A genetic probe encoding a virulence gene from Salmonella typhimurium was useful in the detection of Salmonella from feces during an outbreak of salmonellosis at a local dairy. A 3.2-kb BamHI restriction endonuclease fragment of the S. typhimurium virulence plasmid, pStSR100, has been useful as a DNA probe for both detection of Salmonella sp. and characterization of virulence plasmids from numerous field isolates. This virA probe hybridizes to a highly conserved gene carried on the large virulence plasmids of invasive Salmonella isolates. Colony blots prepared from feces directly plated onto MaConkey's agar failed to detect low numbers of Salmonella sp. However, hybridization of the VirA probe to vacuum blots or colony blots prepared from feces in tetrathionate enrichment broth incubated for 16 hours at 37 C was effective for detecting Salmonella sp. and resulted in an 85.9% correlation with culture results. The probe also demonstrated the highly conserved nature (96%) of the virulence gene among S. cholerae-suis isolate plasmids detected using Southern blot analysis.     

Resistance of Salmonella isolates in Germany

During 2000-2002 the National Veterinary Reference Laboratory for Salmonella (NRL-Salm) in Germany typed 11,911 isolates from animals, food, feed and the environment. All of them were tested for their susceptibility to 17 anti-microbial agents. Sixty-three per cent of all isolates were resistant and 40% were multiresistant (resistant against more than one anti-microbial). This general resistance level was strongly influenced by those specific serotypes which dominate the Salmonella epidemiology in Germany. Salmonella Typhimurium DT104 isolates from pig and cattle, and their resulting food products, were multiresistant in 98 and 94% of the cases respectively. During the period 2000-2003 an increasing quinolone resistance especially in Salmonella isolates from poultry and poultry meat (to 26%) and in S. Paratyphi B D-tartrate positive isolates (to 64%) could be observed. This increase was accompanied by a shift towards higher minimal inhibitory concentrations for ciprofloxacin.     

Detection of Salmonella in faecal, tissue, and feed samples by conventional culture methods and VIDAS Salmonella Test

The VIDAS Salmonella Test (VST) is an enzyme-linked fluorescent immunoassay for the detection of Salmonella-antigens. The suitability of VST for the detection of Salmonella in faecal, tissue, and feed samples was evaluated by the comparison with routine culture methods. From 312 naturally contaminated samples 17 were classified as Salmonella positive by routine methods and 28 by VST. Salmonella were isolated from 15 VST positive samples by the routine method and from eight samples only by an extended culture method. Five positive VST results could not be proved by culture. Two samples were classified as positive by the routine method and as false-negative by VST. The sensitivity varied between 88% and 100% and the specifity between 92% and 100%, depending on the kind of sample. Matrix or serovar specific factors resulting in a false VST result could not be determined. The performance of VST was easy and did not require special experiences. Mostly, samples with Salmonella negative results were faster detected than by culture methods. VST is suitable for the detection of Salmonella in the studied kind of samples especially as a screening method.     

Potential associations between fecal shedding of Salmonella in feedlot cattle treated for apparent respiratory disease and subsequent adverse health outcomes

A prospective cohort study was used to assess whether Salmonella fecal shedding in commercial feedlot cattle treated with antimicrobials for respiratory disease was associated with subsequent adverse health outcomes. Feces were collected per rectum from cattle that were examined for apparent respiratory disease, had a rectal temperature > or = 40 degrees C, and subsequently received antimicrobial treatment. Salmonella were recovered from 918 (73.7%) of 1 245 fecal samples and weekly prevalence estimates ranged from 49 to 100% over the 3-month study. Genotypic and phenotypic characteristics of Salmonella strains in the population were determined. Serogroup E Salmonella were most common (73.3%), followed by C1 (11.0%), C3 (8.6%), and B (1.1%). Predominant serotypes were Orion (46.5%), Anatum (19.8%), Kentucky (8.7%), Montevideo (7.5%), and Senftenberg (4.9%). Few isolates (36/918) were positive for antimicrobial resistance-associated integron gene intI1. Phenotypic susceptibility was associated with isolate intI1 status. Crude re-pull, re-treatment and case fatality risks were higher for cattle that were Salmonella-positive versus -negative at initial treatment, but not statistically different on multivariable analysis. However, case fatality risk was higher for cattle shedding Group B Salmonella than for cattle shedding other serogroups. Lots (groups) with a higher Salmonella prevalence at first treatment had a higher proportion of mortalities occur in a hospital pen, higher overall re-treatment risks, and were more likely to be sampled later in the study. Results indicate a high prevalence of Salmonella in this population of cattle treated for apparent respiratory disease, but that effects associated with clinical outcomes may depend on the Salmonella strain.     

Effects of a commercially available vaccine against Salmonella enterica serotype Newport on milk production, somatic cell count, and shedding of Salmonella organisms in female dairy cattle with no clinical signs of salmonellosis

OBJECTIVE: To determine effects of vaccination with siderophore receptor and porin (SRP) proteins derived from Salmonella enterica serotype Newport on milk production, somatic cell count, and shedding of Salmonella organisms in female dairy cattle. ANIMALS: 180 female Holsteins. PROCEDURES: Cattle were randomly assigned to receive Salmonella Newport SRP vaccine or control solution. Vaccine or control solution was injected 45 to 60 days before parturition, and cattle received a second dose 14 to 21 days before parturition. Milk production was monitored for the first 90 days of lactation. Feces for isolation of Salmonella and blood samples for detection of antibodies against Salmonella Newport were collected at day of first injection and at days 7 to 14 and 28 to 35 of lactation. RESULTS: Cattle inoculated with Salmonella Newport vaccine produced significantly more milk (1.14 kg/d), compared with cattle injected with the control solution. Cattle administered the vaccine had significantly higher concentrations of circulating antibody against Salmonella Newport SRP proteins at 7 to 14 days and 28 to 35 days of lactation. Salmonella Newport was not recovered; however, Salmonella enterica serotype Agona was recovered from 31 (20.3%) cattle, but likelihood of recovery did not differ significantly between vaccinates and control cattle. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of a vaccine against Salmonella Newport SRP proteins to healthy dairy cattle prior to parturition increased milk production, even in cattle without detectable shedding of Salmonella Newport or clinical signs of salmonellosis. Additional research is needed to clarify the mechanisms by which productivity was improved.     

Isolation of Salmonella organisms from the mesenteric lymph nodes of horses at necropsy.

OBJECTIVES: To determine the prevalence of Salmonella infections in horses at necropsy. DESIGN: Cross-sectional prevalence survey. ANIMALS: 102 horses. PROCEDURE: Mesenteric lymph nodes were collected from horses that were necropsied. Horses had died or were euthanatized because of severe disease or at the request of the owner. Twenty-eight of the horses were racehorses euthantized following acute catastrophic injuries on the racetrack. Mesenteric lymph nodes were submitted for Salmonella culture via direct plating of tissue specimens on MacConkey agar and by use of 4 enrichment culture techniques that used tetrathionate and selenite enrichment broth and brilliant green and Salmonella-Shigella selective plating media. RESULTS: Salmonella typhimurium was isolated from the mesenteric lymph nodes of 2 foals (2/102, 1.96% of the horses). Salmonella organisms were not isolated from the mesenteric lymph nodes of adult horses. CONCLUSIONS AND CLINICAL RELEVANCE: Prevalence of Salmonella infections in horses of our study (1.96%) suggests that the results of cross-sectional surveys, using bacteriologic culture to determine prevalence of Salmonella infection, should be interpreted with caution. Prevalence of Salmonella infections determined in a single facility may not reflect the prevalence of Salmonella-infected horses in the general population; furthermore, obtaining a Salmonella isolate from a horse does not establish that the horse is a chronic Salmonella carrier.     

Evaluation of an immunochromatography strip assay for the detection of Salmonella sp. from poultry.

The present study was conducted to evaluate the sensitivity and specificity of an immunochromatography-based diagnostic kit for Salmonella. The analytical sensitivity of the test when using pure colonies of different Salmonella species was in the range of 1 X 10(4) to 1 x 10(5) colony-forming units per milliliter. The strip detected 19 of 22 strains of Salmonella spp. but failed to detect S. worthington, S. choleraesuis var. kunzendorf and S. johannesburg. The strip did not detect 27 different enteric bacteria, including Escherichia coli O157:H7, Campylobacter jejuni, Shigella sonnei, and Vibrio parahaemolyticus. In direct testing of feces (n = 66) from chickens infected with Salmonella typhimurium, the strip had a sensitivity of 12.3% and a specificity of 100%. Evaluation of the strip assay (n = 510) after sample pre-enrichment in 2% buffered peptone water (BPW) yielded a sensitivity of 93.8% and specificity of 89% when compared to isolation and identification with xylose-lysine-tergitol 4 (XLT4) selective plating media. Subsequent enrichment in Hajna tetrathionate (TT) broth yielded a higher sensitivity (94.7%) and specificity (96.8%). The agreement (kappa) between the strip test and isolation was 0.004 in direct fecal testing, 0.82 in BPW, and 0.89 in TT broth. The assay could detect Salmonella sp. as early as 18-48 hours during pre-enrichment and enrichment compared to isolation on XLT4, which required an overnight incubation step for the presumptive isolation and identification of Salmonella.     

Evaluation of herd sampling for Salmonella isolation on midwest and northeast US dairy farms

Epidemiologic investigations of Salmonella infections in dairy cattle often rely on testing fecal samples from individual animals or samples from other farm sources to determine herd infection status. The objectives of this project were to evaluate the effect of sampling frequency on Salmonella isolation and to compare Salmonella isolation and serogroup classification among sample sources on 12 US dairy farms sampled weekly for 7-8 weeks. Three herds per state were enrolled from Michigan, Minnesota, New York and Wisconsin based upon predefined herd-size criteria. Weekly samples were obtained from cattle, bulk tank milk, milk filters, water and feed sources and environmental sites. Samples were submitted to a central laboratory for isolation of Salmonella using standard laboratory procedures. The herd average number of cattle fecal samples collected ranged from 26 to 58 per week. Salmonella was isolated from 9.3% of 4049 fecal samples collected from cattle and 12.9% of 811 samples from other sources. Serogroup C1 was found in more than half of the samples and multiple serogroups were identified among isolates from the same samples and farms. The percentage of herd visits with at least one Salmonella isolate from cattle fecal samples increased with overall herd prevalence of fecal shedding. Only the three herds with an average fecal shedding prevalence of more than 15% had over 85% of weekly visits with at least one positive fecal sample. The prevalence of fecal shedding from different groups of cattle varied widely among herds showing that herds with infected cattle may be classified incorrectly if only one age group is tested. Testing environmental sample sources was more efficient for identifying infected premises than using individual cattle fecal samples.     

Simulation model estimates of test accuracy and predictive values for the Danish Salmonella surveillance program in dairy herds

The Danish government and cattle industry instituted a Salmonella surveillance program in October 2002 to help reduce Salmonella enterica subsp. enterica serotype Dublin (S. Dublin) infections. All dairy herds are tested by measuring antibodies in bulk tank milk at 3-month intervals. The program is based on a well-established ELISA, but the overall test program accuracy and misclassification was not previously investigated. We developed a model to simulate repeated bulk tank milk antibody measurements for dairy herds conditional on true infection status. The distributions of bulk tank milk antibody measurements for infected and noninfected herds were determined from field study data. Herd infection was defined as having either >or=1 Salmonella culture-positive fecal sample or >or=5% within-herd prevalence based on antibody measurements in serum or milk from individual animals. No distinction was made between Dublin and other Salmonella serotypes which cross-react in the ELISA. The simulation model was used to estimate the accuracy of herd classification for true herd-level prevalence values ranging from 0.02 to 0.5. Test program sensitivity was 0.95 across the range of prevalence values evaluated. Specificity was inversely related to prevalence and ranged from 0.83 to 0.98. For a true herd-level infection prevalence of 15%, the estimate for specificity (Sp) was 0.96. Also at the 15% herd-level prevalence, approximately 99% of herds classified as negative in the program would be truly noninfected and 80% of herds classified as positive would be infected. The predictive values were consistent with the primary goal of the surveillance program which was to have confidence that herds classified negative would be free of Salmonella infection.     

乳胶凝集试验检测牛羊淋巴结中的沙门氏菌

采用沙门氏菌多价血清致敏乳胶并制成乳胶抗体,建立沙门氏菌乳胶凝集试验检测方法(LAT),分别用LAT法和常规分离培养检测法对38份牛淋巴结、101份羊淋巴结进行检测.结果:乳胶凝集试验牛淋巴结阳性21份,羊淋巴结阳性44份;常规分离培养检测法牛淋巴结阳性22份,羊淋巴结阳性48份,两种方法的阳性符合率均超过80%.试验表明乳胶凝集试验操作简便、快速、敏感性高、特异性强且可用于现场检测,是一种适合基层单位检测沙门氏菌的可靠方法.     

Diversity of Salmonella serovars in feedyard and nonfeedyard playas of the Southern High Plains in the summer and winter

OBJECTIVE: To compare Salmonella isolates cultured from feedyard and nonfeedyard (control) playas (ie, temporary shallow lakes) of the Southern High Plains. SAMPLE POPULATION: Water and muck (sediment) samples were obtained from 7 feedyard playas and 3 nonfeedyard playas in the winter and summer. PROCEDURE: Each water and muck sample was enriched with sulfur-brilliant-green broth and incubated in a shaker at 37 degrees C for 24 hours. A sample (100 mL) of the incubated bacterial-enriched broth was then mixed with 100 mL of fresh sulfur-brilliant-green enrichment broth and incubated in a shaker at 37 degrees C for 24 hours. After the second incubation, a swab sample was streaked on differential media. Suspect Salmonella isolates were further identified by use of biochemical tests, and Salmonella isolates were confirmed and serovar determinations made. RESULTS: Salmonella isolates were not recovered from the 3 control playas. Seven Salmonella enterica serovars were isolated from 5 of 7 feedyard playas in the summer, and 13 S. enterica serovars were isolated from 7 of 7 feedyard playas in the winter. In the summer, 296 isolates were cultured, and 47 were Salmonella organisms. In the winter, 288 isolates were cultured, and 171 were Salmonella organisms. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that feedyard playas are frequently contaminated with many Salmonella serovars. These pathogens should be considered whenever feedyard managers contemplate the use of water from these playas. Water from feedyard playas should not be used to cool cattle in the summer or for dust abatement.     

Effects of sample storage and delayed secondary enrichment on detection of Salmonella spp in swine feces

OBJECTIVE: To determine effects of fecal sample storage and delayed secondary enrichment (DSE) on detection of Salmonella spp in swine feces. Sample Population-Fecal samples obtained from 84 pigs in a commercial herd. PROCEDURE: Each fecal sample underwent 3 storage treatments: no storage (ie, processed on the day of collection), storage at 4 C for 6 days, and storage at -15 C for 14 days. After assigned storage treatments, all samples were enriched in Rappaport-Vassiladias (RV) broth (single enrichment) and plated on XLT4 agar. Delayed secondary enrichment was performed, using single enrichment broths that were stored for 4 days at room temperature. RESULTS: Of 504 cultures, 186 (36.9%) were Salmonella positive. A difference in proportions of samples with positive results was not found between same-day processing and storage at 4 C for 6 days. Compared with use of single enrichment for 24 hours (34% positive), use of DSE resulted in a greater proportion (40%; P < 0.001) of samples with positive results. Estimated relative sensitivities for the storage methods were 0.90, 0.85, and 0.71 for same-day processing, storage at 4 C for 6 days, and storage at -15 C for 14 days, respectively. CONCLUSIONS: Where practical, processing of fecal samples on the day of collection is recommended, although storage at 4 C for several days does not result in marked loss of sensitivity. Improved detection associated with DSE warrants further investigation and optimization.     

Use of delayed secondary enrichment for the isolation of Salmonella in poultry and poultry environments

A conventional method of isolating Salmonella was compared with isolation using novobiocin-supplemented plating media and delayed secondary enrichment (DSE). The DSE greatly increased the ability to isolate Salmonella from poultry and environmental samples. Four hundred sixty-four isolations of Salmonella were made from a total of 4377 cultures (11%). Two hundred sixty-nine (58%) isolations of Salmonella were made following the 24-hour incubation; of these, 43 (9%) isolates were isolated only at this time. In comparison, a total of 421 (91%) Salmonella were isolated by DSE, of which 195 isolates (42%) were isolated only with DSE. The addition of novobiocin to the selective plating medium increased the isolation rate for Salmonella and reduced the level of contaminating bacteria growing on the plate.     

Isotype-specific antibody responses of cattle to Salmonella Dublin lipopolysaccharide and porin following Salmonella Dublin vaccination and acute and chronic infection.

Stimulation of different T-cell subsets during antigen presentation influences the antibody isotype response to an antigen. Salmonella infection and Salmonella bacterin vaccination are likely to stimulate different T-cell subtypes. The objective of this study was to determine whether there are differences in the isotype response of cattle to Salmonella antigens following Salmonella infection and Salmonella bacterin vaccination. Sera from Salmonella bacterin-vaccinated, experimentally infected, and chronically infected (carrier) adult cattle collected during previous studies was used to evaluate the IgG1, IgG2, and IgM isotype responses of cows to Salmonella serotype Dublin lipopolysaccharide (LPS) and porin. Following vaccination and experimental oral infection, IgG1 titers to LPS and porin rose more quickly and persisted longer than did IgG2 titers. In contrast to Salmonella infection, bacterin vaccination stimulated a weak response to Salmonella porin. Salmonella infection also induced a higher IgG2:IgG1 titer ratio to LPS than did bacterin vaccination. Chronic Salmonella infection induced the highest LPS and porin IgG2:IgG1 titer ratios and the highest correlation between LPS and porin titers. Response operating characteristic curves for each isotype-specific enzyme-linked immunosorbent assay (ELISA) were determined to evaluate the effect of isotype on the sensitivity and specificity of Salmonella ELISA serology for distinguishing sera of Salmonella carriers from those of vaccinated and acutely infected cows. IgG2 titers to LPS and porin provide a more specific indicator of chronic Salmonella infection status than do IgG1 titers to the same antigens with little to no loss in sensitivity.     

Ring-trial results for the cultural detection of Salmonella in poultry faeces

The Directive 2003/99/EG of the European Parliament and of the Council on the monitoring of zoonoses and zoonotic agents demands a quality management (QM) system for the execution of its monitoring programmes. Consequently the National Salmonella Reference Laboratory of Germany performed two ring-trials in 2005 and 2006 on the microbiological detection of Salmonella from poultry feces among all participating laboratories in the Federal States. Salmonella detection was performed according to the EN ISO 6579:2002 standard method which was modified according to the recommendations of the Community Reference Laboratory for Salmonella in Bilthoven, The Netherlands. This method uses modified-semisolid Rappaport-Vassiliadis Agar as the only selective enrichment. In 2005 twenty-four and in 2006 twenty-two laboratories participated.They received eight identical samples of the contamination levels L0 (no Salmonella), L1 (11 and 16 cfu per 10 g faeces respectively) and L2 (292 and 418 cfu per 10 g faeces respectively). For both years the data of 20 laboratories could statistically be evaluated. The relative accuracy of the respected results increased from 88.8% in 2005 to 98% in 2006. This is as well reflected in the improved COR- and Kappa-Indices. Taken all together the data show, that the modified-semisolid Rappaport-Vassiliadis protocol is a sensitive, established method for the detection of Salmonella from poultry faeces.