U.S. Department of Agriculture: Agricultural Research Service, Lincoln, Nebraska


Date of this Version


Document Type



Published in Journal of Applied Microbiology (2007) 103: 1657-1668. DOI:10.1111/j.1365-2672.2007.03405.x


Aim: To develop and validate high throughput methods for the direct enumeration of viable and culturable Salmonella and Escherichia coli O157:H7 in ground beef, carcass, hide and faecal (GCHF) samples from cattle. Methods and

Results: The hydrophobic grid membrane filtration (HGMF) method and the spiral plate count method (SPCM) were evaluated as rapid tools for the estimation of pathogen load using GCHF samples spiked with known levels of Salmonella serotype Typhimurium. Validation studies showed that for a single determination of each sample type the low end of the detection limits were approx. 2·0 × 100 CFU g-1 for ground beef, 5·0 × 10-1 CFU (100 cm2)-1 for Salmonella and 8·0 × 10-1 CFU (100 cm2)-1 for E. coli O157:H7 on carcasses, 4·0 × 101 CFU (100 cm2)-1 for hide and 2·0 × 102 CFU g-1 for faecal samples. In addition, ground beef (n = 609), carcass (n = 1520) and hide (n = 3038) samples were collected from beef-processing plants and faecal samples (n = 3190) were collected from feed-lot cattle, and these samples were tested for the presence of Salmonella and E. coli O157:H7 by enrichment and enumeration methods.

Conclusions: The direct enumeration methods described here are amenable to high throughput sample processing and were found to be cost-effective alternatives to other enumeration methods for the estimation of Salmonella and E. coli O157:H7, in samples collected during cattle production and beef processing.

Significance and Impact of the Study: Use of the methods described here would allow for more routine testing and quantification data collection, providing useful information about the effectiveness of beef processing intervention strategies.