Reveal 8-Hour Test System for detection of Escherichia coli O157:H7 in raw ground beef, raw beef cubes, and iceberg lettuce rinse: collaborative study.

Five different food types were analyzed by the Reveal for E. coli O157:H7 8-Hour Test System (Reveal 8) and either the U.S. Food and Drug Administration's Bacteriological Analytical Manual (BAM) culture method or the U.S. Department of Agriculture Food Safety Inspection Service (FSIS) culture method for the presence of E. coli O157:H7. A total of 27 laboratories representing academia and private industry in the United States and Canada participated. Food types were inoculated with E. coli O157:H7 at 2 different levels: a high level where predominantly positive results were expected, and a low level where fractional recovery was anticipated. During this study, 1,110 samples and controls were analyzed by both the Reveal 8 and by BAM or FSIS by each of the collaborators (2,220 samples in total). For each set of samples, 740 were artificially inoculated with E. coli O157:H7, and 370 were uninoculated controls. The Reveal 8 detected 528 presumptive positives of which 487 were confirmed positive by the BAM culture method. In comparison, BAM and FSIS detected 489 of the 740 artificially contaminated samples as positive. In an additional in-house study performed only on chilled and frozen raw ground beef, 240 artificially inoculated samples were analyzed by both the Reveal 8 and by FSIS. The Reveal 8 detected and confirmed 104 samples as positive compared to 79 confirmed positive by FSIS.

Comparison of the Reveal 20-hour method and the BAM culture method for the detection of Escherichia coli O157:H7 in selected foods and environmental swabs: collaborative study.

Four different food types along with environmental swabs were analyzed by the Reveal for E. coli O157:H7 test (Reveal) and the Bacteriological Analytical Manual (BAM) culture method for the presence of Escherichia coli O157:H7. Twenty-seven laboratories representing academia and private industry in the United States and Canada participated. Sample types were inoculated with E. coli O157:H7 at 2 different levels. Of the 1,095 samples and controls analyzed and confirmed, 459 were positive and 557 were negative by both methods. No statistical differences (p <0.05) were observed between the Reveal and BAM methods.

Reveal for Salmonella test system.

The Reveal for Salmonella (RSS) test system is a presumptive qualitative test that detects the presence of Salmonella organisms in foods within 21 h total testing time, allowing the user to release negative products 24 h earlier than when using other rapid test kits. Foods are enriched with a proprietary resuscitation medium called Revive and then selectively enriched with either Selenite Cystine or Rappaport-Vassiliadis selective media. The enriched culture is used to inoculate the RSS detection device, which initiates a lateral flow through a reagent zone containing anti-Salmonella antibodies conjugated to colloidal gold particles that capture antigens present in the culture. The antigen-antibody complex migrates farther and is captured by an additional anti-Salmonella antibody, causing the colloidal gold to precipitate and form a visual line, indicating a positive result. A procedural control line also will form regardless of the presence of Salmonella organisms to indicate the test is working properly. Existing AOAC Official Methods for Salmonella organisms require a 48 h enrichment before testing. Hence, a food product has to be held before release, adding extra cost to the company and the consumer. The RSS test system was evaluated by quantitative spiking studies. Although AOAC encourages inclusion of naturally contaminated foods, almost all microbiological AOAC validation studies have been performed with artificially contaminated foods for absolute control over the study. The RSS test system is designed to test many food types for Salmonella organisms and has a limit of detection of 5-10 colony-forming units (cfu)/25 g with a false-negative rate of < 1% and a false-positive rate of < 5.0%. It showed an 81% overall agreement with the traditional procedure of the U.S. Department of Agriculture's Food Safety Inspection Service.

Natural Occurrence of Fumonisins in Rice with Fusarium Sheath Rot Disease.

Twenty samples of rough rice (Oryza sativa) (unpolished kernels) collected during the 1995 harvest season from Arkansas (seven samples) and Texas (13 samples) were obtained from rice fields known to include plants with symptoms of Fusarium sheath rot putatively caused by Fusarium proliferatum. Samples were analyzed for fumonisin B1 (FB1) at three laboratories using three different extracting solvents by high-performance liquid chromatography (HPLC) or enzyme-linked immunosorbent assay (ELISA) methods. Forty percent of the samples were positive for FB1 at levels ≤4.3 µg/g by HPLC. The same samples contained FB1 at ≤3.6 µg/g when measured by an ELISA method. Most samples that were positive for FB1 were positive for fumonisin B2 (FB2) and fumonisin B3 (FB3) by HPLC at levels ≤1.2 µg/g. Very good agreement was obtained among the two laboratories using HPLC methods and the third using ELISA. Shelling of the unpolished rice results in hull and brown rice fractions. In a sample that contained 4.3 µg/g in whole kernels, the fumonisin level was very high in hulls (≤16.8 µg/g) and low in brown rice (≤0.9 µg/g). Milling of brown rice results in bran and white rice fractions. Fumonisins were found in bran at a level of ≤3.7 µg/g but were below the level of detection by HPLC in white rice. The presence of fumonisins (FB1, FB2, and FB3) was confirmed by fast atom bombardment/mass spectrometry. This is the first report of fumonisins in naturally contaminated rice in the United States.

Fumonisins Veratox. A new rapid quantitative ELISA for determination of fumonisin in food and feed.

Polyclonal antibodies against fumonisin B1 were produced by immunizing sheep with fumonisin B1-keyhole limpet hemocyanin as an immunogen. A quantitative competitive enzyme-linked immunosorbent assay was developed whereby free fumonisins or sample extract containing fumonisins and enzyme-labelled fumonisin competed for binding to the solid phase-bound antibodies. The color intensity of wells, formed by substrate reaction with the enzyme, was inversely related to FB1 concentration. Detection limits for the assay were 0.1 ng/mL fumonisin B1 and concentrations of fumonisins B1, B2, and B3 required for 50% binding inhibition were 5.5, 23 and 18 ng/mL, respectively. For food and feed analyses, samples were extracted with 70% methanol and dilution of the extracts were used directly for ELISA. ELISA results were compared to HPLC analyses by a reference laboratory and the correlation (r value) between ELISA and HPLC was 0.967. The assay may be used to quantitate fumonisins in food and feed within 30 minutes.

Pelvic osteotomy for bladder exstrophy.

Twenty-five patients with bladder exstrophy underwent pelvic osteotomy at or before initial bladder closure and anterior abdominal repair. Ten patients underwent bilateral iliac osteotomies before 1977. An alternative procedure consisting of bilateral superior pubic ramotomies has been used in 15 patients since 1977. In all patients, successful tension-free closure of the abdomen was achieved. Long-term follow-up of these two groups of patients shows no difference in the degree of pubic diastasis, which was asymptomatic. Posterior iliac osteotomy requires two additional incisions, longer operative time, and postoperative immobilization. We therefore recommend bilateral superior pubic ramotomy as an alternative procedure in initial reconstruction of the anterior abdomen in patients with bladder exstrophy.

Posterior leg sepsis associated with pyogenic arthritis of the knee. A report of two cases.

The association of posterior leg sepsis with pyogenic arthritis of the knee is rare, but nevertheless the diagnosis should be kept in mind. Calf swelling and tenderness with apparent antibiotic resistance in pyogenic arthritis of the knee are the major signs of the diagnosis. Ultrasound, computerized axial tomography, or aspiration of the posterior compartment may show fluid in the posterior compartment. Clinical examination is the most dependable method of diagnosis. Treatment consists of adequate surgical drainage of the posterior compartment and appropriate antibiotic therapy.

Percutaneous embolization of bony pelvic neoplasms with tissue adhesive.

Eight patients with tumors of the bony pelvis underwent embolization with isobutyl-2-cyanoacrylate (IBCA). Five patients had primary bone tumors, of which 2 were malignant and 3 were benign; 3 patients had metastases to the bony pelvis from the thyroid gland, kidney, and femur, respectively. Embolization was performed to minimize blood loss during resection of a giant-cell tumor in one patient and insertion of a hip prosthesis in another who had metastatic renal carcinoma. It was also done prior to scheduled surgery in one of the patients with aneurysmal bone cyst, but healing was sufficient to cancel the operation; in the other patient, embolization was the only therapy. Palliative embolization was performed in 4 patients with malignant tumors after other means failed to control pain or slow progression. IBCA appears to be an efficient means of occluding the vessels feeding selected primary bone tumors and metastases.

Carpal and tarsal osteolysis.

A 19-year-old girl with carpal and tarsal osteolysis has been presented. The clinical course was characterized by normal early growth and motor development followed by the insidious onset in early childhood of a progressive, peripheral joint destruction, especially in the tarsal and carpal bones. The patient's course was complicated by acquired spasticity from platybasia, scoliosis, generalized muscle weakness, mild growth failure and corneal clouding.