This is the first report on the use of a normally lethal dose of ciprofloxacin in a Campylobacter agar medium to kill all ciprofloxacin-sensitive Campylobacter spp. but allow the selective isolation and quantitation of naturally occurring presumptive ciprofloxacin-resistant Campylobacter CFU in rinses from retail raw chicken carcasses (RTCC). Thermophilic-group total Campylobacter CFU and total ciprofloxacin-resistant Campylobacter CFU (irrespective of species) were concurrently quantified in rinses from RTCC by direct plating of centrifuged pellets from 10 or 50 ml out of 400-ml rinse subsamples concurrently on Campylobacter agar and ciprofloxacin-containing Campylobacter agar at 42°C (detection limit ‫؍‬ 0.90 log 10 CFU/carcass).

In 2005, the FDA withdrew approval for the use of fluoroquinolones in live poultry production. To assess any changes in countable numbers of ciprofloxacin-resistant Campylobacter before and after the fluoroquinolone withdrawal, retail whole raw chicken carcasses (RTCC) purchased in Northwest Arkansas from 2004 to 2006 were sampled for this purpose. Using a previously published direct-plating method developed in our laboratory, we quantified trends of Campylobacter and ciprofloxacin-resistant Campylobacter loads by direct plating whole chicken carcass rinses on Campylobacter agar (CA) or Campylobacter agar containing 8.6 mg/ml ciprofloxacin (CCA). Countable populations of Campylobacter were recovered from 74, 96, and 63% of carcasses sampled in 2004, 2005, and 2006 respectively. The percentages of carcasses with minimum detectable levels of ciprofloxacin-resistant Campylobacter were 20, 42 and 33%, sampled in 2004, 2005 and 2006, respectively. Our 3 year analysis in one geographical area indicated a persistence of Campylobacter and ciprofloxacin-resistant Campylobacter on retail raw chicken carcasses despite the cessation of fluoroquinolone use in poultry production.

A total of 10 ciprofloxacin-sensitive (ciprofloxacin minimum inhibitory concentration, MIC < 0.5 micro g/ml) and 10 ciprofloxacin-resistant (MIC 16 to 32 micro g/ml) presumptive C. jejuni were further characterized and evaluated for their inhibition by natural orange oil fractions. Partial species identification was performed by using a hippuricase gene-based polymerase chain reaction (PCR) assay. One of the isolates appeared to be atypical and failed to hydrolyze hippurate. Of the ciprofloxacin-resistant C. jejuni isolates tested, six were found to have their quinolone resistance determined by a C --> T mutation in codon 86 of gyrA. Both groups of ciprofloxacin-sensitive and -resistant C. jejuni isolates were most susceptible to cold-pressed terpeneless Valencia orange oil (C4) which yielded inhibition zones from 44.0 +/- 1.4 to 80 +/- 0.0 mm. Less inhibitory responses were recorded for 5-fold concentrated Valencia orange oil (C3) and distilled d-limonene (C7) which exerted similar effects on both ciprofloxacin-sensitive and -resistant C. jejuni isolates. In general, ciprofloxacin-resistant and -sensitive C. jejuni isolates were equally susceptible to the respective orange oil fractions.

The VIDAS UP Listeria (LPT) is an automated rapid screening enzyme phage-ligand based assay for the detection of Listeria species in human food products and environmental samples. The VIDAS LPT method was compared in a multi-laboratory collaborative study to AOAC Official Method 993.12 Listeria monocytogenes in Milk and Dairy Products reference method following current AOAC guidelines. A total of 14 laboratories participated, representing government and industry, throughout the United States. One matrix, queso fresco (soft Mexican cheese), was analyzed using two different test portion sizes, 25 and 125 g. Samples representing each test portion size were artificially contaminated with Listeria species at three levels, an uninoculated control level [0 colony-forming units (CFU)/test portion], a low-inoculum level (0.2-2 CFU/test portion), and a high-inoculum level (2-5 CFU/test portion). For this evaluation, 1800 unpaired replicate test portions were analyzed by either the VIDAS LPT or AOAC 993.12. Each inoculation level was analyzed using the Probability of Detection (POD) statistical model. For the low-level inoculated test portions, difference in collaborator POD (dLPOD) values of 0.01, (-0.10, 0.13), with 95% confidence intervals, were obtained for both 25 and 125 g test portions. The range of the confidence intervals for dLPOD values for both the 25 and 125 g test portions contains the point 0.0 indicating no statistically significant difference in the number of positive samples detected between the VIDAS LPT and the AOAC methods. In addition to Oxford agar, VIDAS LPT test portions were confirmed using Agar Listeria Ottavani and Agosti (ALOA), a proprietary chromogenic agar for the identification and differentiation of L. monocytogenes and Listeria species. No differences were observed between the two selective agars. The VIDAS LPT method, with the optional ALOA agar confirmation method, was adopted as Official First Action status for the detection of Listeria species in a variety of foods and environmental samples.