Immunological properties of an artificial antigen possessing the specificity of Salmonella O-factor 3

For the first time the method of obtaining synthetic protein-free antigen with the specificity of Salmonella O-factor by the radical copolymerization of the synthetic trisaccharide 3-0 [4-0-(beta-D-mannopyranosyl)-alpha-L-rhamnopyranosyl]-beta-allyl-D- galactopyranoside with acrylamide was developed. The synthetic antigen thus obtained possessed the narrow specificity of serogroup E Salmonella O-factor 3. The serological activity of the antigen, studied in the precipitation and passive hemagglutination tests, was considerably higher than that of lipopolysaccharides isolated from S. anatum and S. newington. This synthetic antigen proved to be nontoxic and possessed immunogenic properties, inducing the formation of antibodies to Salmonella O-factor 3 in immunized animals.     

Thein vitro phagocytic activity of guinea-pig macrophages toSalmonella typhi andSalmonella enteritidis

The engulfing, bactericidal and degrading activities toSalmonella typhi, strain ty2-4446 and 0-901 and toSalmonella enteritidis of guinea pig macrophages obtained from peritoneal exudate, spleen and bone marrow that were cultivated for 2–7 days, were studied. The phagocytic activity was expressed as a total number of phagocytosed microbes and the number of viable bacteria, released from mechanically disrupted macrophages. The ratio of phagocytosed bacteria to the original number of bacteria that were introduced to macrophage cultures, were evaluated in per cents. No significant difference in phagocytic activity was found between macrophages submitted to thein vitro cultivation and macrophages freshly isolated from the organism. Profound variations in phagocytic activity of cells were found which were partially dependent on the dose of microbes employed for the infection of cultures. Furthermore, both the engulfing and bactericidal activity of peritoneal macrophages toSalmonella typhi were found to be higher than in bone morrow macrophages.Salmonella typhi 0-901 microbes were phagocytosed by macrophages from bone marrow and peritoneal exudate much better thanSalmonella typhi ty2. In addition, a significant delay in bactericidal activity toSalmonella typhi ty2 of bone marrow macrophages in comparison to peritoneal macrophages was observed. The spleen macrophages possessed better phagocytic and killing activity toSalmonella enteritidis than bone marrow macrophages. A striking difference was found as regards the intracellular growth ofSalmonella typhi andSalmonella gertneri: no multiplication ofSalmonella typhi within the peritoneal and bone marrow macrophages was observed during the 3–5 h cultivation, whereas on the other hand,Salmonella gertneri started to grow intracellularly within the 5 h cultivation in the bone marrow macrophages.     

Retrospective and longitudinal study of salmonellosis in captive wildlife in Trinidad

Morbidity and mortality of captive wildlife at the Emperor Valley Zoo, Trinidad from 1993 to 1996 were analysed to determine involvement of Salmonella spp. A 6 mo longitudinal study was conducted to determine the frequency of isolation of Salmonella spp. from apparently healthy, sick and dead wild mammals, birds, and reptiles. The antibiograms of Salmonella isolates were determined using the disc diffusion method. Fecal samples randomly selected from animal enclosures and cloacal swabs of snakes were cultured for Salmonella spp. following enrichment in tetrathionate and selenite cystine broths. For the 1993-96 period, Salmonella spp. was implicated in 17 (12%) of 141 sick or dead animals and the predominant serotype was S. typhimurium. During the 6 mo prospective study in a mean animal population of 1,186, there were 20 (2%) and 14 (1%) animals that were sick and died respectively; Salmonella spp. was implicated in only one mortality. Overall, of 1,012 samples from apparently healthy wildlife cultured, 66 (7%) yielded 24 serotypes of Salmonella. The predominant serotype were S. seigburg (16 isolates), S. gaminara (6 isolates), and S. thompson (6 isolates). None of the samples yielded S. typhimurium. The frequency of isolation of Salmonella spp. in reptiles (14%) was significantly higher than found in either mammals (7%) or birds (3%). Sixty-five (99%) of 66 Salmonella spp. isolates exhibited resistance to one or more of the nine antimicrobial agents tested. Resistance was high to cephalothin (92%), moderate to streptomycin (35%) and tetracycline (29%), but significantly low to gentamicin (2%), chloramphenicol (0%), and sulphamethoxazole/trimethoprim (0%). The prevalence of asymptomatic infections by Salmonella spp. in zoo animals was high and the very high prevalence of antimicrobial resistance could be a problem when treating salmonellosis.     

Assessment of the genotoxic effects of methyl chloride in human lymphoblasts

The activity of methyl chloride was measured in 4 genotoxicity assays. In an established human lymphoblast line, a 3-h treatment with 0-5% methyl chloride resulted in a dose-related increase in mutant fraction at the thymidine kinase locus and induction of sister-chromatid exchange. No increase in DNA damage, as measured by alkaline elution, was detected in the lymphoblasts at concentrations of methyl chloride shown to be mutagenic. Also, a concentration-related increase in 8-azaguanine-resistant fraction in Salmonella typhimurium was observed following a 3-h treatment with atmospheres containing 0-20% methyl chloride. Thus, methyl chloride is a weak, direct-acting mutagen for bacteria and human cells in culture.     

2018-2022年佛山地区小儿沙门氏菌感染的临床特征和耐药性分析

摘要 目的：分析2018-2022年佛山地区小儿沙门氏菌感染的临床特征和耐药性,为小儿沙门氏菌感染的临床防治提供依据。方法：对2018年1月1日至2022年12月31日期间佛山市妇幼保健院收集的小儿沙门氏菌感染病例186例进行回顾性分析,分析患儿一般资料、小儿沙门氏菌感染的时间分布特征、临床特征、血清型分布及耐药性。结果：186例小儿沙门氏菌感染患儿年龄分布,1~3岁构成比最高,占37.63%,>6岁构成比最低,占1.61%。2018年发病33例,2019年发病19例,2020年发病39例,2021年发病44例,2022年发病51例,沙门氏菌感染病例数总体呈现上升趋势;一年四季均有沙门氏菌感染病例,夏秋两季发病率最高,分别占43.55%、30.11%。发热、腹泻、解粘液便为主要症状,分别占90.32%、74.73%和63.98%。血清型B群最高,为118例,占63.44%。通过对分离出的191株沙门氏菌药物敏感性试验分析,只有3株（1.57%）对所检测抗菌药物均不耐药,耐3类或3类以上的多重耐药沙门氏菌有157株（82.20%）,庆大霉素、头孢唑啉、头孢替坦耐药率均为100%。不同年份沙门氏菌对左氧氟沙星、环丙沙星、四环素的耐药率差异有统计学意义（P<0.05）。结论：近年来佛山地区小儿沙门氏菌感染呈上升趋势,1~3岁儿童是主要群体,夏季和秋季是发病的高峰季节,主要血清型是B群,儿童感染后以发热、腹泻、解粘液便为主要症状,沙门氏菌的耐药性较为严重,应引起相关部门重视。     

Growth and genetic responses of Salmonella Typhimurium to pH-shifts in an anaerobic continuous culture

Salmonella infection of chickens that leads to potential human foodborne salmonellosis continues to be a concern. Changes in the pH of poultry gastrointestinal tract could influence Salmonella growth and virulence response. In the current study, growth responses of a chicken isolate Salmonella enterica serovar Typhimurium (ST) to three incremental pH-shifts (6.17-7.35) in continuous cultures (CC) were evaluated. The expression of rpoS and hilA was determined by real time-polymerase chain reaction (RT-PCR) as well. Increases in pH resulted in higher cell protein concentrations, glucose disappearance, and glucose and ATP yields. Although with some inconsistency between the two trials, the data indicated that the ammonia release into media was favored by low pH. The pH shifts did not significantly affect acetate biosynthesis. No consistent trends of pH influence on propionate and butyrate production could be detected. In all three pH shifts, relative expression of hilA was dominant at 0h which represented CC steady state. In pH shift 7.35-6.86 (Trial 1), the relative expression of rpoS at time 0 and 1h were over five-fold higher than after 3 and 6h of growth. Overall, the results suggest that ST physiology is altered by changes in pH, which could be determinant factors for ST survival in the poultry gastrointestinal ecosystems.     

Evidence for distinct polygenic regulation of antibody responses to some unrelated antigens in lines of mice selected for high or low antibody responses to somatic antigen of Salmonella

The effect of the selective breeding of mice for high or low antibody production to complex immunogens is largely nonspecific, since it modifies the responsiveness of high (H) and low (L) lines to many antigens unrelated to the selection antigen. However, the nonspecific effect of the polygenic control operating in these lines is not a general feature. For example, the group of genes in selection IV, carried out for responsiveness to somatic antigen of Salmonella, does not modify the responses to sheep erythrocytes (SE). Despite equivalent responses in H and L mice of selection IV, a large variability was found in individual responses of F₂ interline hybrids, which demonstrates the presence of alleles with high or low effect on responses to SE. A selective breeding (Selection IV-A) was therefore initiated from this F₂ population for responsiveness to SE. A progressive interline divergence occurred during the first seven generations of selection; the interline separation was due to polygenic regulation (about four independent loci from a preliminary estimate).Equivalent responses to the s antigen of Salmonella are observed in the two lines. This constitutes additional evidence for distinct polygenic regulation of responses to SE and to somatic antigen. Moreover, the pattern of responses to several unrelated antigens (nonspecific effect) also differs between Selections IV and IV-A.Abbreviations H high responder lines - L low responder lines - s somatic antigen of Salmonella - f flagellar antigen of Salmonella - R response to selection - S selection differential - F₀ foundation population - h² heritability (realized) - RGG rabbit gamma globulin - CE chicken erythrocyte - HE human erythrocyte - PE pigeon erythrocyte - SE sheep erythrocyte     

Reduction of indicator and pathogenic microorganisms by psychrophilic anaerobic digestion in swine slurries

The objective of this study was to evaluate the efficiency of a low temperature anaerobic treatment to reduce viable populations of indicator microorganisms (total coliforms, Escherichia coli) and the presence of selected pathogens (Salmonella, Yersinia enterocolitica, Cryptosporidium and Giardia) in swine slurries from different sources. Experiments were carried out in 40 l Sequencing Batch Reactors (SBRs). Experimental results indicated that anaerobic digestion of swine manure slurry at 20 degrees C for 20 days in an intermittently fed SBR: (1) reduced indigenous populations of total coliforms by 97.94-100%; (2) reduced indigenous populations of E. coli by 99.67-100%; (3) resulted in undetectable levels of indigenous strains of Salmonella, Cryptosporidium, and Giardia. It can be considered as a promising method for reducing indigenous indicator and pathogenic microorganisms populations in liquid swine manure slurries.     

Survival of Escherichia coli O157:H7 and Salmonella typhimurium inoculated on chicken by aqueous chlorine dioxide treatment

Inactivation of Escherichia coli O157:H7 and Salmonella typhimurium was evaluated on inoculated chicken by aqueous chlorine dioxide (ClO2) treatment. Chicken samples were inoculated with 6-7 log CFU/g of Escherichia coli O157:H7 and Salmonella typhimurium, respectively. The chicken samples were then treated with 0, 50, and 100 ppm of ClO2 solution and stored at 4 +/- 1 degrees C. Aqueous ClO2 treatment decreased the populations of the pathogenic bacteria on the chicken breast and drumstick. In particular, 100 ppm ClO2 treatment on the chicken breast and drumstick reduced Escherichia coli O157:H7 and Salmonella typhimurium by 1.00-1.27 and 1.37-1.44 log CFU/g, respectively. Aqueous ClO2 treatment on the growth of the bacteria was continuously in effect during storage, resulting in the decrease of the populations of Escherichia coli O157:H7 and Salmonella typhimurium. These results suggest that aqueous ClO2 treatment should be useful in improving the microbial safety of chicken during storage.     

Comparison of two different types of competitive exclusion products

The efficacy of two different types of commercial competitive exclusion (CE) products against Salmonella was studied in three chicken assay trials. Chicks were treated on the day of hatch and challenged one day later either with Salm. infantis (Trials 1 and 2) or a combination of Salm. infantis and Salm. enteritidis (Trial 3). The caeca of the birds were examined for Salmonella five days after challenge. The mean logarithmic counts of Salmonella were from 3·4 to 5·7 in the groups treated with product A derived from the whole caecal contents of an adult bird, from 0·0 to 1·2 in the groups treated with product B, a highly selected product that does not contain any clostridia, and from 6·3 to 7·6 in the control groups. None of the challenge organisms superseded the other in Trial 3, and neither did the double challenge affect the protective capacity of the treatment materials.     

Mutagenicity of some substituted 1-phenyl-3,3-dimethyltriazenes. I. The salmonella/mammalian microsome assay and repair test

The mutagenic activity and related biological properties of Br-, Cl-, NO2- and CH3-derivatives of 1-(phenyl)-3,3-dimethyltriazene were investigated in Salmonella/microsome assays with standard and preincubation metabolic activation and in the repair test using Salmonella and E. coli B/r. In the repair test, the CH3-derivative was slightly positive in the E. coli recA and uvrA repair system, the NO2-derivative had a killing effect on Salmonella typhimurium uvrB-deficient strains. In Salmonella mutagenicity assays, all tested triazene derivatives reverted frameshift tester strains, especially TA1537. The highest number of frameshift mutations was induced by the CH3-derivative in the presence of a standard metabolic activation system; direct mutagenicity of this derivative was weak, reaching about the same level of activity as seen after preincubation. The only test compound that induced mutations of the base-substitution type was the NO2-derivative; this derivative showed the highest mutagenicity when activated by preincubation.     

鸡白痢沙门菌ipaJ基因的克隆与鉴定

摘要：【目的】从鸡白痢沙门菌C79-13中克隆ipaJ基因,体外表达该蛋白后进行免疫原性分析。【方法】鸡白痢沙门菌C79-13与肠炎沙门菌50041进行抑制差减杂交后获得的片段PEA2、PE31和PE44与猪霍乱沙门菌C500疫苗株pSFD10质粒上ipaJ基因高度同源,拼接后获得了鸡白痢沙门菌完整的ipaJ基因序列。从鸡白痢沙门菌中克隆出ipaJ基因并将其构建到原核表达载体pET-30a(+)上,Western-blot检测体外表达蛋白的免疫原性,同时检测了该基因在鸡白痢沙门菌分离株中的分布。【结果】从鸡白痢沙门菌中克隆了大小为840 bp的ipaJ基因序列,并获得了体外原核表达的大小为37 kDa融合蛋白。该蛋白可与鸡白痢沙门菌阳性血清反应。PCR结果显示该基因广泛存在于鸡白痢沙门菌菌株中。 【结论】本文首次报道和克隆了鸡白痢沙门菌ipaJ基因,并证明了IpaJ蛋白具有免疫原性。     

Analysis of Salmonella enteritidis strains isolated from food-poisoning cases in Taiwan by pulsed field gel electrophoresis, plasmid profile and phage typing

AIMS: To establish the molecular typing data for Salmonella enteritidis due to its increasing role in Salmonella infections in Taiwan. METHODS AND RESULTS: Sixty-three Salm. enteritidis strains isolated from related and unrelated patients suffering from food-borne poisoning during 1991-97 were collected and subjected to pulsed field gel electrophoresis (PFGE), plasmid analysis and phage typing. For PFGE, XbaI, SpeI and NotI restriction enzymes were used for chromosomal DNA digestion. The results showed that, for these 63 Salmonella strains, 10 PFGE pattern combinations were found. Of these, pattern X3 S3 N3 was the major subtype, since 46 strains isolated from different locations at different times during 1991-97 showed this PFGE pattern. Plasmid analysis showed only three plasmid profiles and phage typing showed that most of the Salmonella strains were of the phage type PT4. CONCLUSION: Most of the Salm. enteritidis strains circulating in Taiwan are of very similar genetic types or are highly related and that strains of PFGE pattern X3 S3 N3 are the prevalent and recirculating strains of Salm. enteritidis which caused food-poisoning cases in Taiwan in 1991-97. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides information that in Salmonella infection, certain subtypes of Salm. enteritidis should be scrutinized.     

Operational characteristics of an antibody-immobilized QCM system detecting Salmonella spp

A quartz crystal microbalance (QCM) system detecting Salmonella spp. was developed by an anti-Salmonella antibody immobilization onto one gold surface of a piezoelectric quartz crystal surface with sulfosuccinimidyl 6-[3-(2-pyridyldithio)propionamido]hexanoate (sulfo-LC-SPDP) thiolation. The optimum temperature and pH for the antibody-immobilized sensor were 35 degrees C and 7.2, respectively. The frequency shifts obtained were correlated with the Salmonella concentrations in the range 3.2 x 10(6)-4.8 x 10(8) CFU per ml. The system was quite specific to Salmonella spp. and applicable for repetitive use after a regeneration step employing 1.2 M NaOH. A model sample measurement was done for a market milk spiked with Salmonella typhimurium.     

Influence of temperature fluctuations on Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium in cow manure

The effects of four average temperatures (7, 16, 23 and 33 degrees C) and daily oscillations with three amplitudes (0, +/-4, +/-7 degrees C) on the survival of the enteropathogens Escherichia coli O157:H7 and Salmonella serovar Typhimurium were investigated in small microcosms. Manure was inoculated with a green fluorescent protein transformed strain of either pathogen at 10(7) cells g(-1) dry weight. Samples were collected immediately after inoculation, and 1 and 2 weeks after inoculation for E. coli O157:H7, and immediately and after 2 and 3 weeks for Salmonella serovar Typhimurium. Population densities were determined by dilution plating and direct counting. In addition, total bacterial CFUs were determined. Growth and survival data were fitted to a modified logistic model. Analysis of the estimated parameter values showed that E. coli O157:H7 survived for shorter periods of time and was more sensitive to competition by the native microbial community than Salmonella serovar Typhimurium. Survival of both pathogens significantly declined with increasing mean temperatures and with increasing amplitude in daily temperature oscillations. The results indicated that responses of enteropathogens to fluctuating temperatures cannot be deduced from temperature relationships determined under constant temperatures.     

Pulsed-field gel electrophoresis, plasmid profiles and phage types for the human isolates of Salmonella enterica serovar Enteritidis obtained over 13 years in Taiwan

AIMS: Plasmid profile, phage typing, and pulsed-field gel electrophoresis (PFGE) patterns of 124 Salmonella Enteritidis strains isolated in 1998-2002 in Taiwan were analysed and the results were compared with those of the 63 strains obtained in 1991-1997, so that molecular subtypes and epidemic strains for Salmonella Enteritidis over a 13-year period (1991-2002) could be elucidated. METHODS AND RESULTS: A total of 124 strains of Salmonella Enteritidis isolated from human in Taiwan between 1998 and 2002 were analysed by PFGE, plasmid analysis and phage typing. The results obtained were compared with those of the 63 strains obtained in 1991-1997, so that the clonal relationships for a total of 187 strains obtained over 13 years could be elucidated. For PFGE, restriction enzymes XbaI, SpeI and NotI were used for chromosomal DNA digestion. Results showed 28 PFGE pattern combinations for the 187 Salmonella strains. Of them, pattern X3S3N3 was the major subtype as 130 strains isolated from different locations during 1991-2002 showed this PFGE pattern. For all these 187 strains, the genetic similarity was higher than 80%. Plasmid analysis showed 17 distinct types, which consist of one to four plasmids and the predominant phage type of those strains was PT4 (71.6%) and PT6a (13.4%). The three methods identified different degrees of polymorphism in the following order: plasmid profile (18 types, D = 0.659) > PFGE (28 types, D = 0.512) > phage typing (13 types, D = 0.438). As PFGE patterns, phage type and plasmid profile were combined for subtyping, the 187 strains could be grouped into 46 subtypes and the discriminatory index was raised to 0.795. For these 46 subtypes, the predominant one was X3S3N3/P1/PT4, which contained 77 (41%) isolates. CONCLUSIONS: Most of the Salmonella Enteritidis strains from sporadic cases were with pattern X3S3N3. They were the prevalent and may be the epidemic strains found in Taiwan during 1991-2002. The present study suggested that the several variants were derived from a single clonal line and the genome for strains of Salmonella Enteritidis are highly conserved over a 13-year period (1991-2002). SIGNIFICANCE AND IMPACT OF THE STUDY: The results obtained here are useful for epidemiolgical study of salmonellosis caused by Salmonella Enteritidis in Taiwan. Comparing the data of the present study with those obtained for strains from other countries, the major subtypes for Salmonella Enteritidis infection in the world can be elucidated.     

Molecular dissection of Salmonella-induced membrane ruffling versus invasion

Type III secretion system-mediated injection of a cocktail of bacterial proteins drives actin rearrangements, frequently adopting the shape of prominent protuberances of ruffling membrane, and culminating in host cell invasion of Gram-negative pathogens like Salmonella typhimurium . Different Salmonella effectors are able to bind actin and activate Rho-family GTPases, which have previously been implicated in mediating actin-dependent Salmonella entry by interacting with N-WASP or WAVE-complex, well-established activators of the actin nucleation machine Arp2/3-complex. Using genetic deletion and RNA interference studies, we show here that neither individual nor collective removal of these Arp2/3- complex activators affected host cell invasion as efficiently as Arp2/3-complex knock-down, although the latter was also not essential. However, interference with WAVE-complex function abrogated Salmonella -induced membrane ruffling without significantly affecting entry efficiency, actin or Arp2/3-complex accumulation. In addition, scanning electron microscopy images captured entry events in the absence of prominent membrane ruffles. Finally, localization and RNA interference studies indicated a relevant function in Salmonella entry for the novel Arp2/3-complex regulator WASH. These data establish for the first time that Salmonella invasion is separable from bacteria-induced membrane ruffling, and uncover an additional Arp2/3-complex activator as well as an Arp2/3-complex-independent actin assembly activity that contribute to Salmonella invasion.     

Extragenic Suppressors of Growth Defects in msbB Salmonella

Lipid A, a potent endotoxin which can cause septic shock, anchors lipopolysaccharide (LPS) into the outer leaflet of the outer membrane of gram-negative bacteria. MsbB acylates (KDO)(2)-(lauroyl)-lipid IV-A with myristate during lipid A biosynthesis. Reports of knockouts of the msbB gene describe effects on virulence but describe no evidence of growth defects in Escherichia coli K-12 or Salmonella. Our data confirm the general lack of growth defects in msbB E. coli K-12. In contrast, msbB Salmonella enterica serovar Typhimurium exhibits marked sensitivity to galactose-MacConkey and 6 mM EGTA media. At 37 degrees C in Luria-Bertani (LB) broth, msbB Salmonella cells elongate, form bulges, and grow slowly. msbB Salmonella grow well on LB-no salt (LB-0) agar; however, under specific shaking conditions in LB-0 broth, many msbB Salmonella cells lyse during exponential growth and a fraction of the cells form filaments. msbB Salmonella grow with a near-wild-type growth rate in MSB (LB-0 containing Mg(2+) and Ca(2+)) broth (23 to 42 degrees C). Extragenic compensatory mutations, which partially suppress the growth defects, spontaneously occur at high frequency, and mutants can be isolated on media selective for faster growing derivatives. One of the suppressor mutations maps at 19.8 centisomes and is a recessive IS10 insertional mutation in somA, a gene of unknown function which corresponds to ybjX in E. coli. In addition, random Tn10 mutagenesis carried out in an unsuppressed msbB strain produced a set of Tn10 inserts, not in msbB or somA, that correlate with different suppressor phenotypes. Thus, insertional mutations, in somA and other genes, can suppress the msbB phenotype.     

Plasmid characteristics of Salmonella strains of various origins

Salmonella antibiotic-resistant strains, isolated from patients with hospital infections and from various environmental objects, showed lower virulence than antibiotic-sensitive strains in experiments on mice infected by intraperitoneal and enteral routes. Salmonella strains, sensitive to antimicrobial preparations, contained 1-2 plasmids, while those with multiple drug resistance contained 3-10 plasmids varying in their molecular weight. All these strains, with the exception of one laboratory strain, carried a plasmid with a molecular weight of about 60 Md. A decrease in the virulence of Salmonella strains, carrying R-plasmid, with respect to mice, their natural host, in experimental infection by the above-mentioned routes was probably unrelated to the loss of this plasmid. 80% of Salmonella strains with multiple resistance to antibiotics yielded positive results in the keratoconjunctival and conjunctival tests as compared with 42% of sensitive strains. These data suggest that Salmonella strains, carrying R-plasmid, retained pronounced capacity for local colonization.     

The detection of Salmonella using a combined immunomagnetic separation and ELISA end-detection procedure

The aim of this study was to develop a rapid immunoassay to detect Salmonella bacteria. Skimmed milk powder (SMP) in buffered peptone water was inoculated with six Salmonella strains (Salm. typhimurium, Salm. virchow, Salm. enteritidis, Salm. give, Salm. ealing and Salm. arizonae) at three inoculum levels (about 2-200 cfu 25 g(-1) SMP) and incubated (37 degrees C) overnight. Heat-treated salmonella cells were immobilized on paramagnetic particles and detected within 3 h using the Salmonella genus-specific monoclonal antibody M105 in a microtitre plate based assay. The rapid Salmonella detection method combining immunomagnetic separation and ELISA had a total isolation and detection time of less than 24 h, which is significantly shorter than the conventional techniques requiring 72-96 h. The technique had a sensitivity limit of 10(5)-10(6) cfu ml(-1).