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Automated Immunomagnetic Separation (automate + immunomagnetic_separation)
Selected AbstractsQuantification and prevalence of Salmonella in beef cattle presenting at slaughterJOURNAL OF APPLIED MICROBIOLOGY, Issue 5 2004N. Fegan Abstract Aims:, A survey to determine the prevalence and numbers of Salmonella in beef cattle presented for slaughter at abattoirs across Australia was conducted between September 2002 and January 2003. Methods and Results:, Automated immunomagnetic separation (AIMS) was used for detection and isolation of Salmonella enriched from cattle faeces. Salmonella were enumerated from positive samples using a combination of the Most Probable Number (MPN) technique and AIMS. A total of 310 faecal samples were tested, 155 were from lot-fed cattle and 155 from grass-fed cattle. Salmonella spp. were isolated from 21 (6·8%) of the cattle and the prevalence amongst grass-fed cattle (4·5%) was not significantly different to that found in lot-fed cattle (9%). Counts of Salmonella in positive faeces varied from <3 MPN g,1 of faeces to 2·8 × 103 MPN g,1 and 71% of positive samples had counts <10 MPN g,1 faeces. There was no significant difference in the mean log10 number of Salmonella in faeces of cattle from each production system. Conclusion:, Low numbers of beef cattle were found to shed Salmonella at the time of slaughter and the prevalence and the associated faecal concentrations did not vary significantly with the pre-slaughter production system (grass or lot feeding). The faecal concentration of Salmonella in the majority of faeces was low (<10 MPN g,1) with few high concentrations up to 3 × 103 MPN g,1, suggesting there may be a low risk of carcase contamination. Significance and Impact of the Study:, Beef cattle do not appear to be a major source of entry of Salmonella into the human food chain and the quantitative information contained in this study can be used in quantitative assessments of the associated risk of human salmonellosis. [source] Comparison of the sensitivity of manual and automated immunomagnetic separation methods for detection of Shiga toxin-producing Escherichia coli O157:H7 in milkJOURNAL OF APPLIED MICROBIOLOGY, Issue 6 2002R.D. Reinders Aim:,To determine the sensitivity of methods for detection of injured and uninjured Escherichia coli O157:H7 (E. coli O157) in raw and pasteurized milk. Methods and Results:,Raw milk, pasteurized milk with 1·5% fat content and pasteurized milk with 3·5% fat content were spiked with E. coli O157 at low levels. The samples were enriched in modified tryptone soya broth with novobiocin (mTSBn) at 37°C. Aliquots of the enriched culture were analysed either by manual immunomagnetic separation (MIMS) and culturing on sorbitol MacConkey agar with or without cefixime and potassium tellurite (SMACct or SMAC), or by automated immunomagnetic separation and integrated ELISA (EiaFossÔ). Uninjured E. coli O157 organisms were detected in milk by both methods at 1 cfu 10 ml,1 sample). Injured organisms were detected at levels of about 4 cfu 10 ml,1 sample. Direct enrichment in mTSBn (22 h incubation) showed better sensitivity for injured cells than enrichment in buffered peptone water (BPW, 22 h incubation), or in a two-step enrichment consisting of BPW (6 h, 37°C) and mTSBn (16 h, 37°C), successively. Conclusions:,The methods showed equal sensitivity in that they were both able to detect 1 cfu 10 ml,1 milk sample. Injured organisms can be detected and isolated at a level almost as low as this. A resuscitation step is not recommended for the detection and isolation of injured and non-injured E. coli O157 from milk. Significance and Impact of the Study:,Due to the dilution of contamination in the bulk tank, analysis of milk for the presence of E. coli O157 requires a very sensitive method. Both methods described here are useful for such analysis. [source] Enumeration of Escherichia coli O157 in cattle faeces using most probable number technique and automated immunomagnetic separationLETTERS IN APPLIED MICROBIOLOGY, Issue 1 2004N. Fegan Abstract Aims:, To determine the numbers of Escherichia coli O157 present in the faeces of naturally infected cattle. Methods and Results:, A combination of the most probable number (MPN) technique and automated immunomagnetic separation (AIMS) was used to enumerate E. coli O157 in cattle faeces from both pasture-fed and grain-fed animals. A total of 22 E. coli O157 positive faecal samples were enumerated for E. coli O157 (10 from pasture-fed and 12 from grain-fed animals). The numbers of E. coli O157 in cattle faeces varied from undetectable (<3 MPN g,1 of faeces) to 2·4 × 104 MPN g,1. There was no significant difference (P = 0·06) between the numbers of E. coli O157 in pasture-fed or grain-fed cattle faeces, although the geometric mean (antilog of the mean of log10 transformed MPN values) was higher in grain-fed (130 MPN g,1) than in pasture-fed (13 MPN g,1). Conclusions:, Although the number of samples tested is small, the results indicate that E. coli O157 make up a small proportion of the total E. coli population present in cattle faeces. Significance and Impact of the Study:, Information on the numbers of E. coli O157 present in cattle will assist in developing more robust quantitative risk assessments and formulating intervention strategies. [source] Isolation of Shiga toxin-producing Escherichia coli O103 from sheep using automated immunomagnetic separation (AIMS) and AIMS-ELISA: sheep as the source of a clinical E. coli O103 case?LETTERS IN APPLIED MICROBIOLOGY, Issue 3 2002A.M. Urdahl Aims: To investigate whether a sheep flock was the original reservoir of a Shiga toxin-producing Escherichia coli (STEC) O103 strain causing a clinical human case and to compare the two diagnostic methods automated immunomagnetic separation (AIMS) and AIMS-ELISA. Methods and Results: AIMS detected Escherichia coli O103 in 36·5% of the samples and AIMS-ELISA detected E. coli O103 in 52·1% of the samples. Polymerase chain reaction detected stx1 and eae in three of 109 E. coli O103 isolates. Pulsed field gel electrophoresis showed that the sheep and human STEC O103 were characterized by distinctly different profiles. Conclusions: The sheep flock was shown to carry STEC O103, although an association between the sheep flock and the clinical human case could neither be proven nor eliminated. Substantial agreement was found between AIMS and AIMS-ELISA, but AIMS-ELISA was less time consuming and resulted in a higher recovery of E. coli O103. Significance and Impact of the Study: The study shows that sheep may be carriers of STEC that are associated with human disease and that the methods described can be used to increase the sensitivity of STEC detection. [source] | ||||||||||