Comparison of Three Selective Media and Validation of the VIDAS Campylobacter Assay for the Detection of Campylobacter jejuni in Ground Beef and Fresh-Cut Vegetables

2011 ◽  
Vol 74 (3) ◽  
pp. 456-460 ◽  
Author(s):  
JUNG-WHAN CHON ◽  
JI-YEON HYEON ◽  
IN-SOO CHOI ◽  
CHAN-KYU PARK ◽  
SOO-KI KIM ◽  
...  
Keyword(s):  
Campylobacter Jejuni ◽  
Ground Beef ◽  
Food Samples ◽  
Assay Method ◽  
Enzyme Linked Immunosorbent Assay ◽  
Culture Methods ◽  
Selective Media ◽  
Fresh Cut ◽  
Microaerobic Conditions ◽  
Campylobacter Spp

In this study, three different selective media, modified cefoperazone charcoal deoxycholate agar (mCCDA), Karmali agar, and Preston agar, were compared for isolating Campylobacter jejuni from artificially contaminated ground beef and fresh-cut vegetables that have different levels of background microflora. Concurrently, an automated enzyme-linked immunosorbent assay method for detecting Campylobacter spp. (VIDAS Campylobacter) was evaluated by comparing it with the culture methods. Food samples inoculated with C. jejuni were enriched in Bolton broth at 42°C for 44 h and then streaked onto the three different selective media, followed by incubation under microaerobic conditions at 42°C for 48 h. The enriched Bolton broth (1 ml) was used in the VIDAS Campylobacter assay. No statistical differences in sensitivities were observed between the three selective media for ground beef and fresh-cut vegetables, but the selectivity of Preston agar was better (P < 0.05) than those of mCCDA and Karmali agar. The VIDAS Campylobacter assay showed a recovery rate similar (P > 0.05) to those of all of the medium combinations in ground beef. However, more positive samples (P < 0.05) were detected with the VIDAS Campylobacter than with the selective agars, except for the combinations of mCCDA plus Preston agar or mCCDA plus Karmali agar plus Preston agar in fresh-cut vegetables.

Methods for Recovery of Campylobacter jejuni from Foods

1982 ◽  
Vol 45 (14) ◽  
pp. 1332-1337 ◽  
Author(s):  
NORMAN J. STERN
Keyword(s):  
Campylobacter Jejuni ◽  
Gas Flow ◽  
Raw Milk ◽  
Growth Conditions ◽  
Food Samples ◽  
Human Infection ◽  
Animal Products ◽  
Enrichment Broth ◽  
Selective Media ◽  
Sensitivity And Selectivity

The triangular relationship between Campylobacter jejuni, foods and disease in humans has been well-documented. Many studies have revealed that C. jejuni causes at least as many cases of human gastroenteritis as does Salmonella sp. Foods are an important vehicle in human infection, and raw milk is most frequently implicated. Other animal products also serve as potential sources of infection. C. jejuni has been found on the carcasses of poultry and other domestic animals throughout the world. The organism is microaerophilic and various methods for establishing appropriate growth conditions, such as the Fortner principle, atmosphere replacement and adding of supplements to encourage growth of C. jejuni, are available. Methods developed for use in clinical laboratories lack the necessary sensitivity and selectivity, and therefore have limited use in detecting small numbers of C. jejuni in foods. In one enrichment method for detecting C. jejuni in foods, washings are filtered and centrifuged, the sediment is suspended in the enrichment broth and the suspension is incubated under a constant gas flow at reduced oxygen levels. Following incubation enrichment broth is filtered and plated onto selective media. In another recently developed method, food samples are directly added to an enrichment broth with antibiotics and incubated under a microaerobic atmosphere before selective plating. Butzler's, Skirrow's and Campy-BAP selective media use several antibiotics to which C. jejuni is resistant. The plates are supplemented with horse or sheep blood, depending upon the specific formulation. The optimum temperature for growth of C. jejuni, about 42°C, may also be used for selection. It is now possible to recover 0.1 to 1 cell of C. jejuni per 10 to 25 g of food sample from among 106 to 109 indigenous bacteria. After a characteristic colony is isolated, the key criteria for presumptive identification of C. jejuni by phase-contrast microscopy are darting, corkscrew motion and a comma to spiral shape.

Production and Characterization of a Monoclonal Antibody to Campylobacter jejuni

2009 ◽  
Vol 72 (4) ◽  
pp. 870-875 ◽  
Author(s):  
S. A. HEO ◽  
R. NANNAPANENI ◽  
M. G. JOHNSON ◽  
J. S. PARK ◽  
K. H. SEO
Keyword(s):  
Monoclonal Antibody ◽  
Campylobacter Jejuni ◽  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Campylobacter Coli ◽  
Carbonate Buffer ◽  
Horse Blood ◽  
Microaerobic Conditions ◽  
Immunoglobulin Subclass

Campylobacter species are a group of spiral-shaped bacteria that can cause disease in humans and animals. We developed a high-affinity monoclonal antibody (MAb) probe that recognizes Campylobacter jejuni cells. Cell suspensions grown under microaerobic conditions at 42°C for 20 h on Bolton agar plates with lysed horse blood were used as live and heat-killed preparations, centrifuged at 8,000 × g for 20 min, and resuspended in carbonate buffer (pH 9.6) for coating on the enzyme-linked immunosorbent assay plates. BALB/c mice were immunized with C. jejuni sonicated cells at 107 CFU/ml to generate MAb-producing hybridoma clones. Of about 500 initial hybridoma clones, MAb 33D2, which reacted with C. jejuni and Campylobacter coli, was selected for further evaluation. MAb 33D2 is in the immunoglobulin subclass G2a and had relatively weaker reactivity with the C. coli strains tested. MAb 33D2 did not show any cross-reactions with the nine non-Campylobacter bacteria tested in the enzyme-linked immunosorbent assay and had a stronger affinity for C. jejuni as live versus heat-killed cells. In Western blot assays, MAb 33D2 recognized two major antigens of 62 and 43 kDa in extracts from C. jejuni cells but only one antigen of 62 kDa in extracts from C. coli cells.

scholarly journals USDA FSIS, FDA BAM, AOAC, and ISO Culture Methods BD BBL CHROMagar Listeria Media

2009 ◽  
Vol 92 (4) ◽  
pp. 1105-1117 ◽  
Author(s):  
Vicki Ritter ◽  
Susan Kircher ◽  
Krista Sturm ◽  
Patty Warns ◽  
Nancy Dick
Keyword(s):  
Ground Beef ◽  
Food Samples ◽  
International Organization ◽  
Culture Methods ◽  
Department Of Agriculture ◽  
False Negatives ◽  
Chi Square ◽  
Smoked Salmon ◽  
Inspection Service ◽  
Chi Square Analysis

Abstract BBL CHROMagar Listeria Media (CL) was evaluated for detection of Listeria monocytogenes in raw ground beef, smoked salmon, lettuce, and Brie cheese. The recovery of L. monocytogenes on CL was compared to the U.S. Food and Drug Administration (FDA) Bacteriological Analytical Manual (BAM), U.S. Department of Agriculture (USDA) Food Safety and Inspection Service (FSIS), AOAC, and International Organization for Standardization (ISO) reference-plated media using the recommended pre-enrichments and selective enrichments. Of the 265 food samples tested, 140 were tested using BAM, USDA, or AOAC methods and 125 were tested using ISO methods. CL produced comparable results with the reference methods on all matrixes with a sensitivity of 99.3 and a specificity of 100. No false negatives were found in testing the food matrixes. There was no statistical difference in recovery based on Chi-square analysis. Known isolates were evaluated, and CL had a sensitivity and specificity of 100. The results of this study demonstrate that CL is an effective medium for the recovery and detection of L. monocytogenes in raw ground beef, smoked salmon, lettuce, and Brie cheese using FDA BAM, USDA FSIS, AOAC, and ISO culture methods.

scholarly journals Detection and quantification of Campylobacter in foods: New analytic approaches to detect and quantify Campylobacter spp. in food samples

2020 ◽  
Vol 9 (2) ◽  
Author(s):  
Maria Francesca Peruzy ◽  
Yolande Thérèse Rose Proroga ◽  
Federico Capuano ◽  
Federica Corrado ◽  
Serena Santonicola ◽  
...  
Keyword(s):  
Real Time ◽  
Campylobacter Jejuni ◽  
Internal Control ◽  
Real Time Pcr ◽  
Quantitative Methods ◽  
Bacterial Load ◽  
Digital Pcr ◽  
Food Samples ◽  
Meat Samples ◽  
Campylobacter Spp

The aim of the present study was to develop rapid qualitative and quantitative methods based on the use of Real-Time PCR and Droplet Digital PCR (ddPCR), in order to have reliable techniques to detect and quantify Campylobacter spp. in food samples. The gene 16S-rRNA was used as specific target for Campylobacter spp. Real- Time PCR evaluation assay and a not competitive internal control was ushered in it. To investigate the selectivity of the method, 26 Campylobacter strains and 40 non-Campylobacter strains were tested and in order to verify the application of Real-Time PCR method, 5 pork meat samples were experimentally inoculated with a Campylobacter jejuni strain. Subsequently, dilutions with a bacterial load of Campylobacter jejuni within 10-106 CFU/mL were chosen for the optimization of the ddPCR assay. Lastly, a total of 54 naturally contaminated foods samples were analyzed through molecular (Real-Time PCR and ddPCR) and traditional methods. The Real-Time PCR protocol demonstrated to amplify only the Campylobacter spp. strains and when Campylobacter jejuni was experimentally inoculated in meat samples the pathogen was always detected. The ddPCRs assay allowed to quantify a level of contamination of 10 CFU/mL, but it was unable to quantify levels of 105 – 106 CFU/mL. Lastly, Campylobacter spp. was never detected in the 54 samples tested. In conclusion, the novel analytic approach proposed, based on an initial screening of the samples with Real-Time PCR and then on quantification of Campylobacter spp. with a ddPCR on those positive, represents a quick monitoring tool and, if used correctly, it would allow the implementation of food safety.

scholarly journals The effects of probiotics on Campylobacter jejuni and Salmonella spp. with respect to the meat and the organs of slaughtered chickens

2010 ◽  
Vol 26 (5-6) ◽  
pp. 393-402 ◽  
Author(s):  
S. Ivanovic ◽  
M.Z. Baltic ◽  
N. Karabasil ◽  
S. Lilic
Keyword(s):  
Campylobacter Jejuni ◽  
Bacteriological Examination ◽  
Ambient Conditions ◽  
Perianal Region ◽  
Salmonella Spp ◽  
Selective Media ◽  
Campylobacter Spp

Our research deals with the effects of probiotics on Campylobacter jejuni and Salmonella spp. with respect to the meat and organs of slaughtered chickens. For the scope of our experiment, we used 250 one-day old chicks, divided into 5 groups. Initially, control chicken group was fed with feed not containing probiotics. Other groups were fed with feed containing different probiotics. Fattening-intended food was standardized for all groups. All chicken groups were exposed to the same ambient conditions. Following 42 days period of fattening, chickens were slaughtered. We took 30 samples of liver, intestine and swabs from perianal region for the needs of bacteriological examination. Campylobacter spp. and Salmonella spp. were determined by selective media. On the basis of obtained results, we can say that the application of probiotics in chicken feed reduces considerably the onset of Campylobacter jejuni and Salmonella spp. in meat and organs.

Effect of iron concentration on toxin production in Campylobacter jejuni and Campylobacter coli

1986 ◽  
Vol 32 (5) ◽  
pp. 395-401 ◽  
Author(s):  
Barbara A. McCardell ◽  
Joseph M. Madden ◽  
John T. Stanfield
Keyword(s):  
Campylobacter Jejuni ◽  
Ferrous Sulfate ◽  
Culture Media ◽  
Iron Concentration ◽  
Toxin Production ◽  
Enzyme Linked Immunosorbent Assay ◽  
Campylobacter Coli ◽  
Casamino Acids ◽  
Culture Supernatants ◽  
Campylobacter Spp

The effect of iron concentrations in culture media on supernatant yields of Campylobacter cytotonic toxin (CCT) was studied. Of the 118 Campylobacter spp. strains surveyed, 78.8% produced toxin in brucella broth or in casamino acids – yeast extract (CYE) broth. When the iron concentration of CYE was increased from 0.44 μg/mL (7.9 μM) to 0.65 μg/mL (11.6 μM) by the addition of ferric chloride, 94.9% of the strains were positive for toxin in a ganglioside GM1 based, enzyme-linked immunosorbent assay, using antibody to affinity-purified CCT. The addition of iron as ferrous sulfate was less effective. When four toxin-positive strains were grown in a deferrated medium of conalbumin-treated CYE with 0.04–0.08 μg iron/mL (0.72–1.43 μM), two of the culture supernatants became negative (absorbance at 410 nm, < 0.1 and < 10 ng CCT/mL), and two produced about 90% less CCT but were still classified as positive (absorbance, ≥ 0.1 and ≥ 10 ng CCT/mL). It was therefore concluded that the production of CCT by Campylobacter spp. is influenced by iron concentration.

scholarly journals Waterborne Isolates of Campylobacter jejuni Are Able to Develop Aerotolerance, Survive Exposure to Low Temperature, and Interact With Acanthamoeba polyphaga

2021 ◽  
Vol 12 ◽  
Author(s):  
Ekaterina Shagieva ◽  
Katerina Demnerova ◽  
Hana Michova
Keyword(s):  
Low Temperature ◽  
Campylobacter Jejuni ◽  
Clinical Sample ◽  
Lethal Dose ◽  
Tetracycline Resistance ◽  
Environmental Parameters ◽  
Acanthamoeba Polyphaga ◽  
Culture Methods ◽  
Microaerobic Conditions ◽  
The Impact

Campylobacter jejuni is regarded as the leading cause of bacterial gastroenteritis around the world. Even though it is generally considered to be a sensitive microaerobic pathogen, it is able to survive in the environment outside of the intestinal tract of the host. This study aimed to assess the impact of selected environmental parameters on the survival of 14 C. jejuni isolates of different origins, including 12 water isolates. The isolates were tested for their antibiotic resistance, their ability to survive at low temperature (7°C), develop aerotolerance, and to interact with the potential protozoan host Acanthamoeba polyphaga. The antibiotic susceptibility was determined by standard disk diffusion according to EUCAST. Out of the 14 isolates, 8 were resistant to ciprofloxacin (CIP) and 5 to tetracycline (TET), while only one isolate was resistant to erythromycin (ERY). Five isolates were resistant to two different antibiotic classes. Tetracycline resistance was only observed in isolates isolated from wastewater and a clinical sample. Further, the isolates were tested for their survival at 7°C under both aerobic and microaerobic conditions using standard culture methods. The results showed that under microaerobic conditions, all isolates maintained their cultivability for 4 weeks without a significant decrease in the numbers of bacteria and variation between the isolates. However, significant differences were observed under aerobic conditions (AC). The incubation led to a decrease in the number of cultivable cells, with complete loss of cultivability after 2 weeks (one water isolate), 3 weeks (7 isolates), or 4 weeks of incubation (6 isolates). Further, all isolates were studied for their ability to develop aerotolerance by repetitive subcultivation under microaerobic and subsequently AC. Surprisingly, all isolates were able to adapt and grow under AC. As the last step, 5 isolates were selected to evaluate a potential protective effect provided by A. polyphaga. The cocultivation of isolates with the amoeba resulted in the survival of about 40% of cells treated with an otherwise lethal dose of gentamicin. In summary, C. jejuni is able to adapt and survive in a potentially detrimental environment for a prolonged period of time, which emphasizes the role of the environmental transmission route in the spread of campylobacteriosis.

Effect of non-fat dry milk on recovery of staphylococcal thermonuclease from foods

1979 ◽  
Vol 25 (1) ◽  
pp. 44-46 ◽  
Author(s):  
C. E. Park ◽  
H. B. El Derea ◽  
M. K. Rayman
Keyword(s):  
False Negative ◽  
Ground Beef ◽  
Acid Precipitation ◽  
Food Samples ◽  
Assay Method ◽  
Negative Results ◽  
False Negative Results ◽  
Dry Milk ◽  
Egg Products ◽  
Subsequent Acid

Modification of the method of Tatini et al. (1976) by addition of non-fat dry milk (NFDM) to food samples and subsequent acid precipitation at pH 3.8 enhanced the recovery of staphylococcal thermonuclease (TNase) from most of 37 foods tested. The modified TNase assay method allowed detection of 10 ng (0.002 units) of the enzyme per gram of each of the following foods: ground beef, boiled egg products, whey powder, fruit-containing yogurt, dressings and spreads, potato and egg salads, and pastas, all of which gave false-negative results without NFDM.

Enumeration of Sublethally Injured Escherichia coli O157:H7 ATCC 43895 and Escherichia coli Strain B-41560 Using Selective Agar Overlays versus Commercial Methods

2013 ◽  
Vol 76 (4) ◽  
pp. 674-679 ◽  
Author(s):  
AMANDA R. SMITH ◽  
ALYSHA L. ELLISON ◽  
AMANDA L. ROBINSON ◽  
MARYANNE DRAKE ◽  
SUSAN A. McDOWELL ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Ground Beef ◽  
Food Samples ◽  
Coli Strain ◽  
Milk Formula ◽  
Selective Media ◽  
E Coli ◽  
Infant Milk Formula ◽  
Intervention Measures ◽  
Direct Inoculation

Quality control procedures during food processing may involve direct inoculation of food samples onto appropriate selective media for subsequent enumeration. However, sublethally injured bacteria often fail to grow, enabling them to evade detection and intervention measures and ultimately threaten the health of consumers. This study compares traditional selective and nonselective agar-based overlays versus two commercial systems (Petrifilm and Easygel) for recovery of injured E. coli B-41560 and O157:H7 strains. Bacteria were propagated in tryptic soy broth (TSB), ground beef slurry, and infant milk formula to a density of 106 to 108 CFU/ml and then were stressed for 6 min either in lactic acid (pH 4.5) or heat shocked for 3 min at 60°C. Samples were pour plated in basal layers of either tryptic soy agar (TSA), sorbitol MacConkey agar (SMAC), or violet red bile agar (VRB) and were resuscitated for 4 h prior to addition of agar overlays. Other stressed bacteria were plated directly onto Petrifilm and Easygel. Results indicate that selective and nonselective agar overlays recovered significantly higher numbers (greater than 1 log) of acid-and heat-injured E. coli O157:H7 from TSB, ground beef, and infant milk formula compared with direct plating onto selective media, Petrifilm, or Easygel, while no significant differences among these media combinations were observed for stressed E. coli B-41560. Nonstressed bacteria from TSB and ground beef were also recovered at densities significantly higher in nonselective TSA-TSA and in VRB-VRB and SMAC-SMAC compared with Petrifilm and Easygel. These data underscore the need to implement food safety measures that address sublethally injured pathogens such as E. coli O157:H7 in order to avoid underestimation of true densities for target pathogens.

scholarly journals USDA FSIS, FDA BAM, and ISO Culture Methods BD BBL CHROMagar O157 Media

2009 ◽  
Vol 92 (4) ◽  
pp. 1118-1127 ◽  
Author(s):  
Vicki Ritter ◽  
Susan Kircher ◽  
Nancy Dick
Keyword(s):  
Ground Beef ◽  
Reference Method ◽  
Food Samples ◽  
Culture Methods ◽  
Chi Square ◽  
Apple Cider ◽  
E Coli ◽  
Inspection Service ◽  
Chi Square Analysis ◽  
Method Agreement

Abstract BBL CHROMagar O157 media (CO) was evaluated for detection of Escherichia coli O157:H7 in raw ground beef and unpasteurized apple cider. The recovery of E. coli O157:H7 on CO was compared to the U.S. Food and Drug Administration (FDA) Bacteriological Analytical Manual (BAM), U.S. Department of Agriculture (USDA) Food Safety and Inspection Service (FSIS), and International Organization for Standardization (ISO) reference-plated media using the recommended enrichment broths. Of the 180 food samples tested, 45 were tested using BAM, 45 using the USDA method, and 90 using the ISO method. CO produced comparable results with the reference methods on all matrixes with a sensitivity of 100 and a specificity of 100. No false negatives were found in testing the food matrixes. There was no statistical difference in recovery based on Chi-square analysis. Method agreement for raw ground beef was 85 for the USDA FSIS method and 95 for the ISO method. Method agreement for unpasteurized apple cider was 100 for the ISO and FDA BAM methods. In all cases where method agreement was &lt;100, CO detected more positives than the reference method media. Evaluation of known isolates on CO in inclusivity and exclusivity testing had a sensitivity and specificity of 100. The results of this study demonstrate that CO is an effective medium for the recovery and detection of E. coli O157:H7 in raw ground beef and unpasteurized apple cider using FDA BAM, USDA FSIS, and ISO methods.

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