Matrix Extension Study: Validation of the ANSR®Salmonella Method for Detection of Salmonella spp. in Pasteurized Egg Products

2014 ◽  
Vol 97 (5) ◽  
pp. 1374-1383 ◽  
Author(s):  
Debra Foti ◽  
Lei Zhang ◽  
Preetha Biswas ◽  
Mark Mozola ◽  
Jennifer Rice

Abstract A matrix extension study was conducted to validate the ANSR®Salmonella method for use with pasteurized egg products. Four diverse egg product types were tested by the ANSR method and the U.S. Department of Agriculture's Microbiology Laboratory Guidebook reference culture procedure. There were no significant differences in the number of positive test portions by the two methods for any of the products examined. Independent laboratory testing of pasteurized liquid egg also found no significant difference in performance between the ANSR and reference culture procedures. There were no false-positive results obtained in either internal or independent laboratory testing. Inclusivity testing using a new enrichment medium specifically designed for use with pasteurized egg products produced 112 positive results out of 113 Salmonella spp. strains tested, with only a single strain of S. Weslaco testing negative by the ANSR assay. Exclusivity testing of 38 non-salmonellae produced all negative ANSR results. It is concluded that ANSR Salmonella is a reliable, sensitive, and specific method for detection of Salmonella spp. in pasteurized egg products.

2014 ◽  
Vol 66 (3) ◽  
pp. 641-647
Author(s):  
F.S. Dias ◽  
I.F. Santos ◽  
R.M. Franco ◽  
E.R. Nascimento

Pathogenic microorganisms can reside transiently or permanently in the gallbladder of cattle. Thus, during slaughter, more attention should be given to the gastrointestinal tract, especially to the accessory organ, the gallbladder. The main aim of this study was to characterize the bacterial microbiota present in bile and gallbladder epithelium of cattle slaughtered in a slaughtering plant under sanitary conditions and to evaluate the antimicrobial resistance in strains of the genus Staphylococcus. Thirty intact gallbladders were collected and the in bile and epithelium were researched for the presence of Aerobic Mesophilic Heterotrophic Bacteria (AMHB), Staphylococcusspp., total Enterobacteriaceae, Enterococcus spp. and Salmonella spp. The frequency of isolation of the microorganism mentioned above were, respectively: 23.02%, 14.39%, 13.67%, 24.46%, 0% and 24.46%. Concerning both gallbladder environments, the frequency of isolation of the microorganisms in the epithelium was 64.03%, and in the bile 35.97%, with no statistical difference, but with significant difference between the population averages. In antimicrobial susceptibility testing, strains of Staphylococcusfrom both bile and gallbladder epithelium showed sensitivity to the antimicrobials: penicillin G, ceftriaxone, chloramphenicol and gentamicin. The observation that the gallbladder supports a high frequency of microorganisms brings us to the possible fact that cattle might be a persistent carrier of pathogens of great importance to public health.


2020 ◽  
pp. 108201322094244
Author(s):  
Vesna Đurović ◽  
Mirjana Radovanović ◽  
Leka Mandić ◽  
Desimir Knežević ◽  
Vladimir Zornić ◽  
...  

The aim of this work was to produce biscuits from wheat flour substituted with different amounts of wheat sprout powder (2.5–7.5%). The biscuits were subjected to chemical, phytochemical, and microbial evaluations. The crude protein, fat, and ash contents and the energy value of the biscuits increased with increasing percentage of wheat sprout powder. Adding sprouts resulted in higher values of phenolics, alpha-tocopherol, and antioxidant activity. There was no statistically significant difference in the contents of total phenolics and alpha-tocopherol between biscuits supplemented with 5% sprouts and biscuits substituted with 7.5% sprouts. The phenolic content in biscuits containing 7.5% sprouts was 245 mg gallic acid equivalents (GAE)/100 g dm compared with 110 mg GAE/100 g dm in control biscuits. Antioxidant activity was the highest in biscuits substituted with 7.5% sprouts. All levels of substitution of wheat flour with wheat sprouts had an effect on the nutritional properties of biscuits, but the substitution level of 2.5–5% is recommended for the improvement of their sensorial properties. The biscuits produced had a low microbial load and were microbiologically safe. Escherichia coli, Salmonella spp., and sulfite-reduction clostridia were not detected in any sample during the period of investigation from 2 to 60 days of storage.


2011 ◽  
Vol 94 (4) ◽  
pp. 1106-1116 ◽  
Author(s):  
Priya Balachandran ◽  
Yanxiang Cao ◽  
Lily Wong ◽  
Manohar R Furtado ◽  
Olga V Petrauskene ◽  
...  

Abstract Real-time PCR methods for detecting foodborne pathogens offer the advantages of simplicity and quick time-to-results compared to traditional culture methods. In this study, the MicroSEQ® real-time PCR system was evaluated for detection of Salmonella spp. in 10 different food matrixes following the AOAC Research Institute's Performance Tested MethodSM validation program. In addition, the performance of the MicroSEQ system was evaluated for the detection of Salmonella in peanut butter as a part of the Emergency Response Validation Program sponsored by the AOAC Research Institute. The system was compared to the ISO 6579 reference method using a paired-study design for detecting Salmonella spp. in raw ground beef, raw chicken, raw shrimp, Brie cheese, shell eggs, cantaloupe, chocolate, black pepper, dry infant formula, and dry pet food. For the peanut butter study, the system was compared to the U.S. Food and Drug Administration's Bacteriological Analytical Manual procedures using an unpaired-study design. No significant difference in performance was observed between the MicroSEQ Salmonella spp. detection system and the corresponding reference methods for all 11 food matrixes. The MicroSEQ system detected all Salmonella strains tested, while showing good discrimination against detection of an exclusivity panel of 30 strains, with high accuracy.


2009 ◽  
Vol 92 (2) ◽  
pp. 449-458 ◽  
Author(s):  
Susan Alles ◽  
Linda X Peng ◽  
Mark A Mozola

Abstract A modification to Performance-Tested MethodSM (PTM) 070601, Reveal® Listeria Test (Reveal), is described. The modified method uses a new media formulation, LESS enrichment broth, in single-step enrichment protocols for both foods and environmental sponge and swab samples. Food samples are enriched for 2730 h at 30C and environmental samples for 2448 h at 30C. Implementation of these abbreviated enrichment procedures allows test results to be obtained on a next-day basis. In testing of 14 food types in internal comparative studies with inoculated samples, there was a statistically significant difference in performance between the Reveal and reference culture U.S. Food and Drug Administration's Bacteriological Analytical Manual (FDA/BAM) or U.S. Department of Agriculture-Food Safety and Inspection Service (USDA-FSIS) methods for only a single food in one trial (pasteurized crab meat) at the 27 h enrichment time point, with more positive results obtained with the FDA/BAM reference method. No foods showed statistically significant differences in method performance at the 30 h time point. Independent laboratory testing of 3 foods again produced a statistically significant difference in results for crab meat at the 27 h time point; otherwise results of the Reveal and reference methods were statistically equivalent. Overall, considering both internal and independent laboratory trials, sensitivity of the Reveal method relative to the reference culture procedures in testing of foods was 85.9 at 27 h and 97.1 at 30 h. Results from 5 environmental surfaces inoculated with various strains of Listeria spp. showed that the Reveal method was more productive than the reference USDA-FSIS culture procedure for 3 surfaces (stainless steel, plastic, and cast iron), whereas results were statistically equivalent to the reference method for the other 2 surfaces (ceramic tile and sealed concrete). An independent laboratory trial with ceramic tile inoculated with L. monocytogenes confirmed the effectiveness of the Reveal method at the 24 h time point. Overall, sensitivity of the Reveal method at 24 h relative to that of the USDA-FSIS method was 153. The Reveal method exhibited extremely high specificity, with only a single false-positive result in all trials combined for overall specificity of 99.5.


2001 ◽  
Vol 64 (11) ◽  
pp. 1844-1847 ◽  
Author(s):  
R. L. ALTHAUS ◽  
M. P. MOLINA ◽  
M. RODRIGUEZ ◽  
N. FERNANDEZ

The Penzym is an enzymatic test widely used for the detection of β-lactam antibiotic residuals in milk. It is a specific method with good sensitivity to this group of antibiotics and enables results to be obtained within a short time. In the present work, the detection limits of 10 β-lactam antibiotics were determined in ewe milk, given the lack of previous studies of the Penzym test in ovine milk. For each antibiotic, eight concentrations were tested on 20 ewe milk samples proceeding from individual ewes (160 analyses per drug). The limits of the Penzym test were determined by means of logistic regression models, as follows: 5 μg/kg amoxicillin, 4 μg/kg ampicillin, 33 μg/kg cloxacillin, 3 μg/kg penicillin “G,” 43 μg/kg ce-phadroxil, 10 μg/kg cephalosporin “C,” 16 μg/kg cephalexin, 900 μg/kg cephoperazone, 120 μg/kg Ceftiofur, and 77 μg/kg cephuroxime. The percentages of positive results for those antibiotics at the maximum residue limit (MRL) concentration established by the European Union (EU) were: 100% (penicillin “G”), 93.3% (ampicillin), 93.3% (cloxacillin), 56.7% (Ceftiofur), and 56.7% (amoxicillin).


1973 ◽  
Vol 29 (2) ◽  
pp. 221-228 ◽  
Author(s):  
T. H. Grenby ◽  
Frances M. Paterson ◽  
R. A. Cawson

1. Techniques were devised for the operation of a new small-scale gnotobiotic rat unit.2. The unit was then used to compare the cariogenicity of sucrose and glucose under carefully controlled conditions in the presence of a single strain of a streptococcus, as gnotobiotic experiments by other workers had given conflicting results.3. Streptococcus IB-1600 was implanted into thirty-four rats, which were then fed on high-sugar diets under gnotobiotic conditions for 5 or 8 weeks from weaning. The level of caries was significantly higher on the sucrose than on the glucose diet, but there was no significant difference in the extent of soft coronal dental plaque.


2013 ◽  
Vol 96 (3) ◽  
pp. 532-541
Author(s):  
Olga Petrauskene ◽  
Yan Cao ◽  
Patrick Zoder ◽  
Lily Y Wong ◽  
Priya Balachandran ◽  
...  

Abstract The Applied Biosystems Performance Tested MethodSM for detecting Listeria species in food and environmental samples was compared to the Health Canada reference method (MFHPB-30) for the analysis of five ready-to-eat (RTE) meats (deli turkey, hot dogs, liver paté, deli ham, and raw fermented sausage) and a stainless steel surface. The MicroSEQ method includes the MicroSEQ®Listeria spp. Detection Kit and the option of two different sample preparation kits, either the automated high-throughput PrepSEQ™ Nucleic Acid Extraction Kit or the manual low- to mid-throughput PrepSEQ™ Rapid Spin Sample Preparation Kit. For each sample matrix, 20 replicates were analyzed at two inoculum levels: for RTE meats a low-level inoculum at 0.2–2 CFU/25 g and a high-level inoculum at 2–5 CFU/25 g; and for environmental surfaces, a low-level inoculum at 0.2–2 CFU/5 cm2 sampling area and a high-level inoculum at 2–5 CFU/5 cm2 sampling area. Five control replicates were also analyzed at 0 CFU/25 g (uninoculated) for food or 0 CFU/5 cm2 sampling area for environmental surface. Both sample preparation methods returned identical results. There was no statistically significant difference in the number of positive samples detected by the MicroSEQ Listeria species method and the MFHPB-30 reference method for three RTE meats and for the one stainless steel environmental surface tested. For deli turkey, there was a statistically significant difference in the number of positive results detected by the MicroSEQ method and the Health Canada MFHPB-30 reference method for the low inoculation level, with the MicroSEQ method detecting more positives. For hot dogs, statistical equivalence was not applicable since hot dogs were spiked with 10x Listeria innocua as competitive background, and the MicroSEQ method detects all known Listeria. Because the MicroSEQ method uses real-time PCR to detect pathogens, it provides faster time-to-results with equivalent detection compared to culture methods. The MicroSEQ method detects Listeria species within 2 to 3 h following 24 to 28 h enrichment compared to culture methods that take at least 5 days for presumptive positive results.


2003 ◽  
Vol 66 (7) ◽  
pp. 1146-1153 ◽  
Author(s):  
I. SAULI ◽  
J. DANUSER ◽  
C. WENK ◽  
K. D. C. STÄRK

Various safety assurance measures are implemented in Switzerland throughout the food production chain to prevent foods of animal origin from being contaminated with Salmonella. The data that are generated from the implementation of these measures are dispersed and heterogeneous. This hinders a general overview and makes a comprehensive national evaluation of the safety assurance level difficult. A semiquantitative method that considers the quality and relevance of the various safety assurance measures for Salmonella spp. was developed. The method uses the data generated from the implementation of safety assurance measures on a national basis (gathered by interviewing stakeholders in the production step). By assembling and analyzing the data systematically, the safety assurance level for Salmonella spp. can be evaluated at every step of the food production chain. This method allows the detection of strengths and weaknesses of the safety system. The systematic evaluation procedures permit comparisons between production steps and product categories. The method was used for evaluating the safety assurance levels throughout the production chain of eggs and egg products in Switzerland. Results of the analysis showed that the overall safety assurance levels for Salmonella spp. at all production steps for eggs and egg products were good. The relatively straightforward implementation of the method made it particularly appropriate in the context of a preliminary evaluation. The method does not have the same high level of detail that is provided by microbial quantitative risk assessments, but it allows an analyst to provide meaningful results when the large amount of data required for a quantitative approach are not present while including the entire “farm to fork” continuum. It may be used as a basis for more in-depth assessments of food safety levels within various production sectors. The method could be adapted for evaluating the safety assurance for other zoonotic foodborne pathogens of interest, such as Campylobacter spp.


1999 ◽  
Vol 62 (11) ◽  
pp. 1243-1247 ◽  
Author(s):  
SUSAN E. ANSAY ◽  
KIM A. DARLING ◽  
CHARLES W. KASPAR

The survival of Escherichia coli O157:H7 and of a nonpathogenic control strain of E. coli was monitored in raw ground beef that was stored at 2°C for 4 weeks, −2°C for 4 weeks, 15°C for 4 h and then −2°C for 4 weeks, and −20°C. Irradiated ground beef was inoculated with one E. coli control strain or with a four-strain cocktail of E. coli O157:H7 (ca. 105 CFU/g), formed into patties (30 to 45 g), and stored at the appropriate temperature. The numbers of the E. coli control strain decreased by 1.4 log10 CFU/g, and pathogen numbers declined 1.9 log10 CFU/g when patties were stored for 4 weeks at 2°C. When patties were stored at −2°C for 4 weeks, the numbers of the E. coli control strain and the serotype O157:H7 strains decreased 2.8 and 1.5 log10 CFU/g, respectively. Patties stored at 15°C for 4 h prior to storage at −2°C for 4 weeks resulted in 1.6 and 2.7 log10–CFU/g reduction in the numbers of E. coli and E. coli O157:H7, respectively. Storage of retail ground beef at 15°C for 4 h (tempering) did not result in increased numbers of colony forming units per gram, as determined with violet red bile, MRS lactobacilli, and plate-count agars. Frozen storage (−20°C) of ground-beef patties that had been inoculated with a single strain of E. coli resulted in approximately a 1 to 2 log10–CFU/g reduction in the numbers of the control strain and individual serotype O157:H7 strains after 1 year. There was no significant difference between the survival of the control strain and the O157:H7 strains, nor was there a difference between O157:H7 strains. These data demonstrate that tempering of ground-beef patties prior to low-temperature storage accelerated the decline in the numbers of E. coli O157:H7.


2011 ◽  
Vol 2011 ◽  
pp. 1-4 ◽  
Author(s):  
Gözde Derviş Hakim ◽  
Şafak Kızıltaş ◽  
Hilmi Çiftçi ◽  
Şafak Göktaş ◽  
İlyas Tuncer

Background and Aims. We aimed to investigate the prevalence of Giardiasis in patients with dyspepsia and patients with diabetes mellitus. Methods. 400 patients and 100 healthy persons were included in this clinical prospective study. The number of patients in each group was equal, 200 dyspeptic and 200 diabetic, respectively. The antigen of G. lntestinalis was determined in the stool specimens by ELISA method. Results. The frequency of Giardiasis was 7% in dyspeptic and 15% in diabetic patients. There was no positive results in any of the healthy persons. There was a significant difference in prevalence rate of Giardiasis between patients with dyspepsia and diabetes mellitus (P<0.05). Conclusions. These results revealed that the prevalence of Giardiasis in dyspepsia and with diabetes mellitus was high in our country. This is the first study investigating the prevalence of Giardiasis in diabetic patients. To investigate Giardiasis in diabetic patients, who have dyspepsia or not, may be a good approach for public health.


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