Evaluation of the Culture Method NIHSJ-02 Alternative to ISO 10272-1:2006 for the Detection of Campylobacter jejuni and Campylobacter coli in Chicken: Collaborative Study

2013 ◽  
Vol 96 (5) ◽  
pp. 991-997 ◽  
Author(s):  
Yoshika Momose ◽  
Yumiko Okada ◽  
Hiroshi Asakura ◽  
Tomoya Ekawa ◽  
Kazuya Masuda ◽  
...  
Keyword(s):  
Campylobacter Jejuni ◽  
Qualitative Method ◽  
Collaborative Study ◽  
Reference Method ◽  
Culture Method ◽  
International Organization ◽  
Campylobacter Coli ◽  
Selective Detection ◽  
Low Level ◽  
High Level

Abstract For the surveillance of the prevalence of Campylobacter jejuni and Campylobacter coli in raw chicken products in Japan, a qualitative method, National Institute of Health Sciences Japan (NIHSJ)-02, was developed as an alternative to International Organization for Standardization (ISO) 10272-1:2006. In the NIHSJ-02 culture method, the enrichment step is carried out in a reduced volume of Preston broth at 42 ± 1°C to reduce cost and space, and to prevent the overgrowth of background bacteria. To evaluate the performance of NIHSJ-02, a collaborative study was conducted, and the results obtained by NIHSJ-02 were compared with those obtained using the reference method, ISO 10272-1:2006. Fifteen laboratories participated; each examined 48 minced chicken samples consisting of test samples uninoculated, inoculated with C. jejuni at a low or high level, and inoculated with C. coli at a low level. The average probabilities of detection by NIHSJ-02 across laboratories were 0.033, 0.222, 0.678, and 0.267 in samples uninoculated, inoculated with C. jejuni at a low and high level, and with C. coli at a low level, respectively. Those by ISO 10272-1:2006 were 0.051, 0.128, 0.551, and 0.090. Significantly higher probabilities of detection were determined by NIHSJ-02 compared to ISO 10272-1:2006, except for uninoculated samples. On the other hand, significantly lower frequency of occurrence of background bacteria was observed by NIHSJ-02 (43.1%) compared with ISO 10272-1:2006 (92.6%). NIHSJ-02 showed better performance than ISO 10272-1:2006 with regard to the selective detection of C. jejuni and C. coli in chicken.

Detection of Campylobacter jejuni and Campylobacter coli from Broiler Chicken–Related Samples Using BAX PCR and Conventional International Organization for Standardization Culture

2008 ◽  
Vol 71 (4) ◽  
pp. 835-838 ◽  
Author(s):  
LISA K. WILLIAMS ◽  
ALISDAIR MCMEECHAN ◽  
TAMSIN BAALHAM ◽  
LAURA WARD ◽  
TOM J. HUMPHREY ◽  
...  
Keyword(s):  
Campylobacter Jejuni ◽  
Broiler Chicken ◽  
Culture Method ◽  
International Organization ◽  
Campylobacter Coli ◽  
Poultry Production ◽  
Production Chain ◽  
Enrichment Cultures ◽  
International Organization For Standardization ◽  
Campylobacter Spp

In this study, the conventional International Organization for Standardization (ISO) culture method was compared with the DuPont Qualicon BAX system, a high-throughput, rapid molecular assay that can be used to detect several bacterial species, including Campylobacter jejuni and Campylobacter coli in diverse sample types. Standard enrichment culture is a time-consuming process, taking up to 6 days to obtain a confirmed result. Rapid molecular assays have been developed that provide results within 24 h. Naturally contaminated samples from the poultry production chain were examined for the presence of Campylobacter spp. Samples from broiler chicken ceca (n = 100), fresh chicken carcass rinses (n = 60), and bootsocks (gauze sock walked through a broiler chicken house; n = 50) were enriched according to the ISO 10272 method in Bolton broth specifically designed to detect Campylobacter spp. in complex sample types. Samples were enriched without blood for use with the BAX system using the Campylobacter BAX kits for the detection of C. jejuni and C. coli. Samples also were directly plated onto modified charcoal cefperazone deoxycholate agar, and results were compared with those from the enriched samples for the ability to detect Campylobacter spp. Campylobacter spp. were isolated from 49% of samples with conventional enrichment cultures, from 48% with direct culture, from 68% with the BAX system and enrichment cultures, and from 62% with the BAX system used directly with samples. Overall, the BAX system detected more positive samples than did the conventional culture method and is an effective methodology for the rapid and reliable detection of Campylobacter spp. from diverse sample types.

Role of the plasmid-encoded tet(O) gene in tetracycline-resistant clinical isolates of Campylobacter jejuni and Campylobacter coli

2007 ◽  
Vol 56 (6) ◽  
pp. 833-837 ◽  
Author(s):  
Javid Iqbal Dasti ◽  
Uwe Groß ◽  
Sven Pohl ◽  
Raimond Lugert ◽  
Michael Weig ◽  
...  
Keyword(s):  
Molecular Markers ◽  
Campylobacter Jejuni ◽  
Acute Gastroenteritis ◽  
Tetracycline Resistance ◽  
Gene Localization ◽  
Campylobacter Coli ◽  
Low Level ◽  
High Level ◽  
Level Resistance

The prevalence of tetracycline resistance, tetracycline MICs and tet(O) gene localization were investigated in 83 Campylobacter isolates from patients suffering from acute gastroenteritis in Germany. Combined biochemical and molecular markers identified 74 isolates (89 %) as Campylobacter jejuni, including seven atypical isolates that failed to hydrolyse hippurate, and nine isolates (11 %) as Campylobacter coli. Tetracycline resistance was detected in six out of nine Campylobacter coli isolates (67 %) and 13 out of 74 C. jejuni isolates (18 %). Low-level tetracycline resistance was observed for C. coli (MIC 16 μg ml−1 for all strains), whereas C. jejuni showed high-level resistance (MIC >256 μg ml−1 for all strains). Both low- and high-level tetracycline resistance was associated with the presence of the tet(O) gene. In C. jejuni, tet(O) was plasmid-encoded in 54 % of tetracycline-resistant isolates, whereas in C. coli, tet(O) appeared to be located on the chromosome.

Culture Method for Detection of Salmonella in Dried Active Yeast: Collaborative Study

1976 ◽  
Vol 59 (4) ◽  
pp. 731-733
Author(s):  
Paul L Poelma ◽  
Aida Romero ◽  
Clyde R Wilson ◽  
Wallace H Andrews
Keyword(s):  
Collaborative Study ◽  
Culture Method ◽  
Final Evaluation ◽  
Lauryl Sulfate ◽  
Enrichment Medium ◽  
Low Level ◽  
Detection And Identification ◽  
High Level

Abstract A method for detecting Salmonella in dried active yeast was subjected to collaborative study. This method employs trypticase soy broth as the pre-enrichment medium, a sample-to-broth ratio of 1:10, and subsequent transfers to lauryl sulfate tryptose broth and tetrathionate before streaking onto selective agars. Each collaborating analyst received ten 25 g samples of dried active yeast. Duplicate 25 g samples were each inoculated with Salmonella Oranienburg at a low level (28 cells) and a high level (107 cells). Similarly, duplicate 25 g samples were each inoculated with S. senftenberg at a low level (30 cells) and a high level (114 cells). The remaining 2 of 10 samples were not inoculated. Results from 12 of 13 collaborators were evaluated. Only 2 (8.2%) of the 24 low level S. oranienburg samples were reported incorrectly as negative. Twelve of the analysts detected S. senftenberg at both levels and S. Oranienburg at the high level in the inoculated samples. Results from 12 collaborators used in the final evaluation show that 117 of 119 (98.3%) collaborative determinations are in agreement. The official final action method for the detection and identification of Salmonella, 46.013–46.026, has been revised official first action to include applicability to dried active yeast.

scholarly journals Significant contribution of the CmeABC Efflux pump in high-level resistance to ciprofloxacin and tetracycline in Campylobacter jejuni and Campylobacter coli clinical isolates

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Saeed Sharifi ◽  
Bita Bakhshi ◽  
Shahin Najar-peerayeh
Keyword(s):  
Gene Expression ◽  
Point Mutation ◽  
Campylobacter Jejuni ◽  
Significant Contribution ◽  
Efflux Pump ◽  
Campylobacter Coli ◽  
Rapd Pcr ◽  
Resistant Strains ◽  
High Level ◽  
Level Resistance

Abstract Background Campylobacter resistance to antimicrobial agents is regarded as a major concern worldwide. The aim of this study was to investigate the expression of the CmeABC efflux pump and the RAPD-PCR pattern in drug-resistant Campylobacter isolates. Methods A total of 283 stool specimens were collected from children under the age of five with diarrhea. The minimum inhibitory concentration (MIC) of tetracycline and ciprofloxacin was determined by broth microdilution method and E-test, respectively. Detection of tetracycline and ciprofloxacin determinants was done by amplification of tetO gene and PCR-sequencing of the gyrA gene. The cmeABC transcriptional expression was analyzed by Real-time (RT)-PCR. Clonal correlation of resistant strains was determined by RAPD-PCR genotyping. Results Out of 283 fecal samples, 20 (7.02%) samples were positive for Campylobacter spp. Analysis of duplex PCR assay of the cadF gene showed that 737 and 461 bp amplicons were corresponding to Campylobacter jejuni and Campylobacter coli, respectively. All of the 17 phenotypically tetracycline-resistant Campylobacter isolates harbored the tetO gene. Also, four phenotypically ciprofloxacin-resistant Campylobacter isolates had a point mutation at codon 257 of the gyrA gene (ACA to ATA; Thr > Ile). High-level expression of the cmeA gene was observed in ciprofloxacin-resistant and high-level tetracycline-resistant Campylobacter isolates, suggesting a positive correlation between the cmeA gene expression level and tetracycline resistance level. Moreover, a statistically significant difference was observed in the cmeA gene expression between ciprofloxacin-resistant and ciprofloxacin-susceptible strains, which signifies the crucial contribution of the efflux pump in conferring multiple drug resistance phenotype among Campylobacter spp. RAPD analysis of Campylobacter isolates exhibited 16 different patterns. Simpsone`s diversity index of RAPD-PCR was calculated as 0.85, showing a high level of homogeneity among the population; however, no clear correlation was detected among tetracycline and/or ciprofloxacin resistant isolates. Conclusion Significant contribution of the CmeABC efflux pump in conferring high-level resistance to tetracycline and ciprofloxacin was observed in C. jejuni and C. coli clinical isolates. The resistant phenotype is suggested to be mediated by CmeABC efflux pumps, the tetO gene, and point mutation of the gyrA gene. Genotyping revealed no clonal correlation among resistant strains, indicating distinct evolution of tetracycline and ciprofloxacin resistant genotypes among the isolates.

Enrichment Broth for the Detection of Campylobacter jejuni and Campylobacter coli in Fresh Produce and Poultry

2017 ◽  
Vol 80 (11) ◽  
pp. 1842-1850 ◽  
Author(s):  
Youmi Jo ◽  
Hye-Min Oh ◽  
Yohan Yoon ◽  
Sun-Young Lee ◽  
Ji-Hyoung Ha ◽  
...  
Keyword(s):  
Campylobacter Jejuni ◽  
Fresh Produce ◽  
International Organization ◽  
Natural Food ◽  
Campylobacter Coli ◽  
Standardization Method ◽  
Potassium Tellurite ◽  
Enrichment Broth ◽  
International Organization For Standardization ◽  
Campylobacter Spp

ABSTRACT Although campylobacteriosis caused by Campylobacter jejuni and Campylobacter coli has been increasingly reported worldwide owing to the consumption of contaminated poultry and fresh produce, the current detection protocols are not selective enough to inhibit unspecific microbes other than these pathogens. Five antibiotics were separately added to Bolton broth, and the survival rates of 18 Campylobacter spp. and 79 non-Campylobacter spp. were evaluated. The survival rate of the non-Campylobacter spp. was the lowest in Bolton broth with rifampin (6.3%), followed by cefsulodin (12.7%), novobiocin (16.5%), and potassium tellurite and sulfamethozaxole (both 17.7%). Also the most effective concentration of rifampin was found to be 12.5 mg/L, which markedly inhibited non-Campylobacter strains while not affecting the survival of Campylobacter strains. After the Campylobacter spp. were enriched in Bolton broth supplemented with 12.5 mg/L rifampin (R-Bolton broth), CampyFood Agar (CFA) was found to be better in selectively isolating the pathogens in the enrichment broth than the International Organization for Standardization method of using modified charcoal cefoperazone deoxycholate agar (mCCDA) for this step. When applied to natural food samples—here, romaine lettuce, pepper, cherry tomato, Korean leek, and chicken—the R-Bolton broth–CFA combination decreased the number of false-positive results by 50.0, 4.2, 20.8, 50.0, and 94.4%, respectively, compared with the International Organization for Standardization method (Bolton broth–mCCDA combination). These results demonstrate that the combination of R-Bolton broth and CFA is more efficient in detecting C. jejuni and C. coli in poultry and fresh produce and thus should replace the Bolton broth–mCCDA combination.

scholarly journals 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

2001 ◽  
Vol 84 (3) ◽  
pp. 719-736 ◽  
Author(s):  
Charles B Bird ◽  
Rebecca J Hoerner ◽  
Lawrence Restaino ◽  
G Anderson ◽  
W Birbari ◽  
...  
Keyword(s):  
Collaborative Study ◽  
Ground Beef ◽  
Culture Method ◽  
The United States ◽  
Test System ◽  
Private Industry ◽  
E Coli ◽  
Inspection Service ◽  
High Level ◽  
The U.S

Abstract 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, 1110 samples and controls were analyzed by both the Reveal 8 and by BAM or FSIS by each of the collaborators (2220 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.

scholarly journals Development of a Rapid and Sensitive Method Combining a Cellulose Ester Microfilter and a Real-Time Quantitative PCR Assay To Detect Campylobacter jejuni and Campylobacter coli in 20 Liters of Drinking Water or Low-Turbidity Waters

2011 ◽  
Vol 78 (3) ◽  
pp. 839-845 ◽  
Author(s):  
Adeline Tissier ◽  
Martine Denis ◽  
Philippe Hartemann ◽  
Benoît Gassilloud
Keyword(s):  
Drinking Water ◽  
Real Time ◽  
Quantitative Pcr ◽  
Campylobacter Jejuni ◽  
Tap Water ◽  
Culture Method ◽  
Cellulose Ester ◽  
Campylobacter Coli ◽  
Content Type ◽  
Real Time Quantitative Pcr

ABSTRACTInvestigations ofCampylobacter jejuniandCampylobacter coliin samples of drinking water suspected of being at the origin of an outbreak very often lead to negative results. One of the reasons for this failure is the small volume of water typically used for detecting these pathogens (10 to 1,000 ml). The efficiencies of three microfilters and different elution procedures were determined using real-time quantitative PCR to propose a procedure allowing detection ofCampylobacterin 20 liters of drinking water or low-turbidity water samples. The results showed that more than 80% of the bacteria inoculated in 1 liter of drinking water were retained on each microfilter. An elution with a solution containing 3% beef extract, 0.05 M glycine at pH 9, combined with direct extraction of the bacterial genomes retained on the cellulose ester microfilter, allowed recovery of 87.3% (±22% [standard deviation]) ofCampylobacterper 1 liter of tap water. Recoveries obtained from 20-liter volumes of tap water spiked with aC. colistrain were 69.5% (±10.3%) and 78.5% (±15.1%) for 91 CFU and 36 CFU, respectively. Finally, tests performed on eight samples of 20 liters of groundwater collected from an alluvial well used for the production of drinking water revealed the presence ofC. jejuniandC. coligenomes, whereas no bacteria were detected with the normative culture method in volumes ranging from 10 to 1,000 ml. In the absence of available epidemiological data and information on bacterial viability, these last results indicate only that the water resource is not protected from contamination byCampylobacter.

scholarly journals Antimicrobial susceptibility patterns of thermophilic Campylobacter spp. from humans, pigs, cattle, and broilers in Denmark.

1997 ◽  
Vol 41 (10) ◽  
pp. 2244-2250 ◽  
Author(s):  
F M Aarestrup ◽  
E M Nielsen ◽  
M Madsen ◽  
J Engberg
Keyword(s):  
Campylobacter Jejuni ◽  
Antimicrobial Agents ◽  
Campylobacter Coli ◽  
Low Frequencies ◽  
Thermophilic Campylobacter ◽  
Campylobacter Lari ◽  
High Level ◽  
Different Origins ◽  
Susceptibility Patterns ◽  
Campylobacter Spp

The MICs of 16 antimicrobial agents were determined for 202 Campylobacter jejuni isolates, 123 Campylobacter coli isolates, and 6 Campylobacter lari isolates from humans and food animals in Denmark. The C. jejuni isolates originated from humans (75), broilers (95), cattle (29), and pigs (3); the C. coli isolates originated from humans (7), broilers (17), and pigs (99); and the C. lari isolates originated from broilers (5) and cattle (1). All isolates were susceptible to apramycin, neomycin, and gentamicin. Only a few C. jejuni isolates were resistant to one or more antimicrobial agents. Resistance to tetracycline was more common among C. jejuni isolates from humans (11%) than among C. jejuni isolates from animals (0 to 2%). More resistance to streptomycin was found among C. jejuni isolates from cattle (10%) than among those from humans (4%) or broilers (1%). A greater proportion of C. coli than of C. jejuni isolates were resistant to the other antimicrobial agents tested. Isolates were in most cases either coresistant to tylosin, spiramycin, and erythromycin or susceptible to all three antibiotics. More macrolide-resistant isolates were observed among C. coli isolates from swine (79%) than among C. coli isolates from broilers (18%) and humans (14%). Twenty-four percent of C. coli isolates from pigs were resistant to enrofloxacin, whereas 29% of C. coli isolates from humans and none from broilers were resistant. More resistance to streptomycin was observed among C. coli isolates from swine (48%) than among C. coli isolates from broilers (6%) or humans (0%). The six C. lari isolates were susceptible to all antimicrobial agents except ampicillin and nalidixic acid. This study showed that antimicrobial resistance was found only at relatively low frequencies among C. jejuni and C. lari isolates. Among C. coli isolates, especially from swine, there was a high level of resistance to macrolides and streptomycin. Furthermore, this study showed differences in the resistance to antimicrobial agents among Campylobacter isolates of different origins.

Evaluation of VIDAS® UP Salmonella (SPT) Assay for the Detection of Salmonella in a Variety of Foods and Environmental Samples: Collaborative Study

2013 ◽  
Vol 96 (4) ◽  
pp. 808-821 ◽  
Author(s):  
Patrick Bird ◽  
Kiel Fisher ◽  
Megan Boyle ◽  
Travis Huffman ◽  
Marc Juenger ◽  
...  
Keyword(s):  
Confidence Intervals ◽  
Environmental Samples ◽  
Collaborative Study ◽  
Reference Method ◽  
The United States ◽  
Probability Of Detection ◽  
Inoculum Level ◽  
Test Portion ◽  
Low Level ◽  
Significant Difference

Abstract The VIDAS® UP Salmonella (SPT) uses recombinant phage proteins to detect Salmonella species in human and animal food products and production environmental samples after 18–26 h of enrichment. The VIDAS SPT assay is performed with the automated VIDAS or mini-VIDAS instruments. The VIDAS SPT method was compared in a multilaboratory collaborative study to the U.S. Department of Agriculture/Food Safety and Inspection Service-Microbiology Laboratory Guidebook (USDA/FSIS-MLG) 4.05 (2011) Isolation and Identification of Salmonella from Meat, Poultry, Pasteurized Egg and Catfish Products reference method following the current AOAC guidelines. A total of 15 laboratories representing government, academia, and industry throughout the United States participated. One matrix, raw ground beef, was analyzed using two different test portion sizes, 25 and 375 g. Each test portion was artificially contaminated with Salmonella at three inoculation levels, an uninoculated control level (0 CFU/test portion), a low inoculum level (0.2–2 CFU/test portion), and a high inoculum level (2–5 CFU/test portion). In this study, 1656 unpaired replicate samples were analyzed. Of those unpaired replicates, 476 were presumptive positive by the VIDAS method, with 475 confirmed positive by the traditional confirmation procedures and 476 confirmed positive by an alternative confirmation procedure. There were 411 confirmed positive replicates by the USDA/FSIS-MLG reference method. Statistical analysis was conducted according to the probability of detection (POD). For the low-level 375 g test portions, the following dLPOD values, with 95% confidence intervals, were obtained: 0.01 (−0.12, +0.15) for samples confirmed following the traditional confirmation; 0.02 (−0.18, +0.2) for samples confirmed following traditional confirmation on IBISA and ASAP; and 0.03 (−0.18, +0.24) for samples confirmed following the alternative confirmation on IBISA and ASAP. For the low-level 25 g test portions, the following dLPOD values, with 95% confidence intervals, were obtained: 0.41, (0.32, +0.49) for samples confirmed following the traditional confirmation, the traditional confirmation on IBISA and ASAP, and the alternative confirmation on IBISA and ASAP. With 0.0 within the confidence intervals for the 375 g test portions, there was no statistically significant difference in the number of positive samples detected by the VIDAS SPT method and the USDA/FSIS-MLG method at the 0.05 level. For the 25 g test portions, a statistically significant difference was observed between the VIDAS SPT method and the reference method for the low inoculum level, where the VIDAS SPT method recovered a higher number of positive results than the reference method. It is recommended that the VIDAS SPT method with the optional ASAP and IBISA agar confirmation method be adopted for Official First Action status for the detection of Salmonella in a variety of foods and environmental samples.

High Level of Antimicrobial Resistance in Campylobacter jejuni Isolated from Broiler Chickens in Estonia in 2005 and 2006

2007 ◽  
Vol 70 (8) ◽  
pp. 1940-1944 ◽  
Author(s):  
MATI ROASTO ◽  
KADRIN JUHKAM ◽  
TERJE TAMME ◽  
ARI HÖRMAN ◽  
LIIDIA HÄKKINEN ◽  
...  
Keyword(s):  
Multidrug Resistance ◽  
Antimicrobial Resistance ◽  
Campylobacter Jejuni ◽  
Broiler Chickens ◽  
Animal Origin ◽  
Campylobacter Coli ◽  
National Veterinary Institute ◽  
Food Of Animal Origin ◽  
Meat Samples ◽  
High Level

The development of antimicrobial resistance in Campylobacter jejuni and Campylobacter coli is a matter of increasing concern. Because campylobacteriosis is transmitted to humans usually via food of animal origin, the presence of antimicrobial-resistant campylobacters in broiler chickens has important public health implications. The aim of our study was to analyze resistance patterns of C. jejuni isolated from fecal samples collected at a large Estonian chicken farm, from cecal contents collected at slaughterhouses, and from meat samples collected at the retail establishments in 2005 and 2006. A total of 131 C. jejuni isolates were collected over a 13-month period and tested by the broth microdilution VetMIC method (National Veterinary Institute, Uppsala, Sweden) to determine the MICs of various antimicrobials. Resistance to one or more antimicrobials was detected in 104 (79.4%) of the 131 isolates. High proportions of the isolates were resistant to enrofloxacin (73.3%) and nalidixic acid (75.6%). Multidrug resistance (resistance to three or more unrelated antimicrobials) was detected in 36 isolates (27.5%), all of which were resistant to enrofloxacin. Multidrug resistance was significantly associated with enrofloxacin resistance (P < 0.01), and the use of enrofloxacin may select for multiresistant strains.

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