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.

Development of a Novel Chromogenic Medium for Improved Campylobacter Detection from Poultry Samples

2015 ◽  
Vol 78 (9) ◽  
pp. 1750-1755 ◽  
Author(s):  
HAJIME TERAMURA ◽  
MIHOKO IWASAKI ◽  
HIROKAZU OGIHARA
Keyword(s):  
Escherichia Coli ◽  
Campylobacter Jejuni ◽  
Campylobacter Coli ◽  
Chromogenic Medium ◽  
Enrichment Broth ◽  
Selective Media ◽  
E Coli ◽  
Significant Difference

The presence of expanded-spectrum β-lactamase (ESBL)–producing Escherichia coli is a common problem in the isolation of Campylobacter from poultry samples using conventional cefoperazone-based selective media. A novel chromogenic medium (CM-HT), based on modified charcoal cefoperazone deoxycholate agar (mCCDA), has been developed as a solution for improved Campylobacter detection from poultry samples. Although the basic components of CM-HT are the same as mCCDA, CM-HT uses both granular charcoal and sodium cefoxitin to enhance viewability and inhibit ESBL-producing bacteria. All tested Campylobacter jejuni (n = 31) and Campylobacter coli (n = 6) strains grew and formed purple-colored colonies on CM-HT. In contrast, the growth of all other tested microorganisms, including ESBL-producing E. coli strains, was suppressed by this medium. Additionally, 84 poultry samples were examined for the presence of Campylobacter using the ISO 10272-1 method (enrichment with Bolton broth) and the NIHSJ-02 method (enrichment with Preston broth) with mCCDA and CM-HT media for the isolation. The numbers of samples from which Camplylobacter was detected on CM-HT using Preston and Bolton broth were 22 and 18, whereas the numbers on mCCDA were 22 and 13, respectively. Only Campylobacter was detected on CM-HT using both enrichment broths; however, there were 5 and 19 samples from which ESBL-producing E. coli was detected on mCCDA using Preston and Bolton broth, respectively. Thus, there was a significant difference between CM-HT and mCCDA in selectivity for ESBL-producing E. coli regardless of which enrichment broth was used. The results obtained demonstrated that CM-HT is a possible solution for the improved isolation of Campylobacter from poultry samples.

A New Method for Arcobacter butzleri, Arcobacter cryaerophilus, and Arcobacter skirrowii Detection using a Novel Chromogenic Agar

2020 ◽  
Author(s):  
Paul Nguyen ◽  
Oscar Juárez ◽  
Lawrence Restaino
Keyword(s):  
Detection System ◽  
Optimal Growth ◽  
Growth Conditions ◽  
Food Samples ◽  
Test Method ◽  
Good Growth ◽  
Enrichment Broth ◽  
Selective Enrichment ◽  
Arcobacter Butzleri ◽  
Selective Enrichment Broth

Arcobacter species are Gram-negative rods that have been implicated in food- and waterborne illness. Although various cultural isolation methods have been proposed, the current procedures are unable to fully suppress growth of background microbiota present in food samples which inhibits Arcobacter isolation. The purpose of this study was to develop a selective enrichment broth and chromogenic plating medium to detect three Arcobacter species that have been recognized as emerging foodborne pathogens: Arcobacter butzleri , Arcobacter cryaerophilus and Arcobacter skirrowii . The developed Nguyen-Restaino-Juárez Arcobacter detection system consists of a selective enrichment broth (NRJ-B) and a selective/differential plating media (NRJ-M). The protocol of the detection method was determined by evaluating growth of A. butzleri , A. cryaerophilus and A. skirrowii under various temperature (30, 35 and 42ᴼC) and incubation (aerobic, microaerophilic and anaerobic) conditions. Additionally, 47 Arcobacter strains and 39 non- Arcobacter strains were tested in the inclusivity and exclusivity evaluations of NRJ-B and NRJ-M. Overall, the study determined the optimal growth conditions of Arcobacter species using the NRJ- Arcobacter detection system was aerobic incubation at 30ᴼC. NRJ-B supported good growth of A. butzleri , A. cryaerophilus , and A. skirrowii while effectively suppressing growth of non- Arcobacter strains after 48 h. Furthermore, NRJ-M yielded 97.8% inclusivity and 100.0% exclusivity using the tested strains and resulted in salmon-pigmented Arcobacter colonies (1.0 to 1.5 mm in diameter) after 72 h. The novel protocol is the first to develop a chromogenic plating media for the isolation of Arcobacter species. This simple and reliable test method would greatly contribute to understanding the distribution of pathogenic Arcobacter species in food samples.

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.

Isolation of Listeria monocytogenes from Food Products on Four Selective Plating Media

1990 ◽  
Vol 53 (5) ◽  
pp. 382-385 ◽  
Author(s):  
N. P. TIWARI ◽  
S. G. ALDENRATH
Keyword(s):  
Listeria Monocytogenes ◽  
Lithium Chloride ◽  
Raw Milk ◽  
Broth Culture ◽  
Processed Meat ◽  
Enrichment Broth ◽  
Selective Media ◽  
Fresh Vegetables ◽  
Selective Plating ◽  
Serotype 4B

The suitability of four selective media for isolation of Listeria monocytogenes strains has been evaluated. Samples of cheese, processed meat, fresh vegetables, and raw milk were inoculated with low numbers of L. monocytogenes cells. Inoculated samples were enriched at 30°C and plated on selective media for isolation. Modified McBride Agar was found to be substantially inferior in comparison with other media. Acriflavine-Ceftazidime Agar was relatively more efficient but was not satisfactory for all classes of foods and all strains of L. monocytogenes. With L. monocytogenes serotype 4b and strain 286, Lithium Chloride-Phenylethanol-Moxalactam Agar (LPM) and Oxford Agar (OA) were approximately equally effective for isolation from all classes of foods. However, LPM performed poorly with L. monocytogenes serotype 3a. OA medium was consistently superior and gave higher recovery with all strains studied. Incubation of enrichment broth culture for just one day was not sufficient and 2 d of incubation was necessary to achieve a satisfactory level of performance.

scholarly journals Gastrointestinal infections caused by consumption of raw drinking milk in England & Wales, 1992–2017

2019 ◽  
Vol 147 ◽  
Author(s):  
N. Adams ◽  
L. Byrne ◽  
J. Edge ◽  
A. Hoban ◽  
C. Jenkins ◽  
...  
Keyword(s):  
Public Health ◽  
Infectious Disease ◽  
Campylobacter Jejuni ◽  
Raw Milk ◽  
National Surveillance ◽  
Gastrointestinal Infections ◽  
Food Standards ◽  
The Uk ◽  
Drinking Milk ◽  
Intestinal Infectious Disease

Abstract Systematic, national surveillance of outbreaks of intestinal infectious disease has been undertaken by Public Health England (PHE) since 1992. Between 1992 and 2002, there were 19 outbreaks linked to raw drinking milk (RDM) or products made using raw milk, involving 229 people; 36 of these were hospitalised. There followed an eleven-year period (2003–2013) where no outbreaks linked to RDM were reported. However, since 2014 seven outbreaks of Escherichia coli O157:H7 (n = 3) or Campylobacter jejuni (n = 4) caused by contaminated RDM were investigated and reported. Between 2014 and 2017, there were 114 cases, five reported hospitalisations and one death. The data presented within this review indicated that the risk of RDM has increased since 2014. Despite the labelling requirements and recommendations that children should not consume RDM, almost a third of outbreak cases were children. In addition, there has been an increase in consumer popularity and in registered RDM producers in the UK. The Food Standards Agency (FSA) continue to provide advice on RDM to consumers and have recently made additional recommendations to enhance existing controls around registration and hygiene of RDM producers.

scholarly journals Genetic Diversity of Campylobacter jejuni Isolates from Farm Animals and the Farm Environment

2003 ◽  
Vol 69 (12) ◽  
pp. 7409-7413 ◽  
Author(s):  
F. M. Colles ◽  
K. Jones ◽  
R. M. Harding ◽  
M. C. J. Maiden
Keyword(s):  
Genetic Diversity ◽  
Campylobacter Jejuni ◽  
Human Disease ◽  
Clonal Complex ◽  
Food Samples ◽  
Complex Model ◽  
Farm Animals ◽  
Significant Genetic Differentiation ◽  
Sequence Types ◽  
Retail Food

ABSTRACT The genetic diversity of Campylobacter jejuni isolates from farm animals and their environment was investigated by multilocus sequence typing (MLST). A total of 30 genotypes, defined by allelic profiles (assigned to sequence types [STs]), were found in 112 C. jejuni isolates originating in poultry, cattle, sheep, starlings, and slurry. All but two of these genotypes belonged to one of nine C. jejuni clonal complexes previously identified in isolates from human disease and retail food samples and one clonal complex previously associated with an environmental source. There was some evidence for the association of certain clonal complexes with particular farm animals: isolates belonging to the ST-45 complex predominated among poultry isolates but were absent among sheep isolates, while isolates belonging to the ST-61 and ST-42 complexes were predominant among sheep isolates but were absent from the poultry isolates. In contrast, ST-21 complex isolates were distributed among the different isolation sources. Comparison with MLST data from 91 human disease isolates showed small but significant genetic differentiation between the farm and human isolates; however, representatives of six clonal complexes were found in both samples. These data demonstrate that MLST and the clonal complex model can be used to identify and compare the genotypes of C. jejuni isolates from farm animals and the environment with those from retail food and human disease.

scholarly journals Genome Sequence and Annotation of a Human Infection Isolate of Escherichia coli O26:H11 Involved in a Raw Milk Cheese Outbreak

2015 ◽  
Vol 3 (1) ◽  
Author(s):  
Wessam Galia ◽  
Patricia Mariani-Kurkdjian ◽  
Estelle Loukiadis ◽  
Stéphanie Blanquet-Diot ◽  
Françoise Leriche ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Genome Sequence ◽  
Raw Milk ◽  
Human Infection ◽  
Raw Milk Cheese

scholarly journals MOLECULAR DETECTION AND CHARACTERIZATION OF STAPHYLOCOCCUS AUREUS ISOLATED FROM RAW MILK SOLD IN DIFFERENT MARKETS OF BANGLADESH

2017 ◽  
Vol 14 (2) ◽  
pp. 277-282 ◽  
Author(s):  
M. A. Islam ◽  
S. M. L. Kabir ◽  
M. T. Rahman
Keyword(s):  
Molecular Detection ◽  
Culture Media ◽  
Raw Milk ◽  
Rrna Gene ◽  
Biochemical Tests ◽  
Selective Media ◽  
Milk Samples ◽  
Antimicrobial Sensitivity ◽  
Antimicrobial Sensitivity Test

The study was intended for molecular detection of S. aureus isolated from raw cow’s milk. A total of 20 milk samples were collected from different upazila markets of Jamalpur, Tangail, Kishoreganj and Netrokona districts of Bangladesh. Milk samples were cultured onto various culture media for the isolation of bacteria. The isolated bacteria were identified by studying cultural properties on different selective media, biochemical tests, and finally by PCR. Out of 20 samples, 15 (75%) milk samples were found to be positive for S. aureus. S. aureus specific 16S rRNA gene was amplified from all isolates and identified as S. aureus. Antimicrobial sensitivity test was carried out to ascertain the susceptibility of the organism to various antibiotics. Its results showed that the S. aureus isolates were resistant to amoxicillin (100%), erythromycin (73.33%) and tetracycline (73.33%) but sensitive to azithromycin (93.33%), ciprofloxacin (93.33%), gentamicin (100%), norfloxacin (86.67%) and streptomycin (86.67%).

scholarly journals Comparison of Genotypes and Antibiotic Resistances of Campylobacter jejuni and Campylobacter coli on Chicken Retail Meat and at Slaughter

2013 ◽  
Vol 79 (12) ◽  
pp. 3875-3878 ◽  
Author(s):  
Sonja Kittl ◽  
Bożena M. Korczak ◽  
Lilian Niederer ◽  
Andreas Baumgartner ◽  
Sabina Buettner ◽  
...  
Keyword(s):  
Campylobacter Jejuni ◽  
Human Infection ◽  
Chicken Meat ◽  
Campylobacter Coli ◽  
Production Chain ◽  
Content Type ◽  
Resistance Patterns ◽  
Retail Meat ◽  
Retail Chicken Meat ◽  
Antibiotic Resistance Patterns

ABSTRACTMultilocus sequence typing (MLST) and antibiotic resistance patterns ofCampylobacter jejuniandCampylobacter colifrom retail chicken meat showed high overlap with isolates collected at slaughterhouses, indicating little selection along the production chain. They also showed significant common sequence types with human clinical isolates, revealing chicken meat as a likely source for human infection.

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