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.

Relative Productivity of Five Selective Plating Agars for the Recovery of Salmonella from Selected Food Types

1979 ◽  
Vol 62 (2) ◽  
pp. 320-326
Author(s):  
Wallace H Andrews ◽  
Clyde R Wilson ◽  
Paul L Poelma ◽  
Aida Romero
Keyword(s):  
Brilliant Green ◽  
Food Samples ◽  
Food Category ◽  
Enrichment Broth ◽  
Selective Enrichment ◽  
Selective Plating ◽  
Relative Productivity ◽  
Selective Enrichment Broth

Abstract During a 3-year period, the relative productivity of brilliant green (BG), bismuth sulfite (BS), Salmonella-Shigella (SS), Hektoen enteric (HE), and xylose lysine desoxycholate (XLD) agars for recovering Salmonella from 9 food types was determined. Following preenrichment, selective enrichment of food samples in tetrathionate broth followed by streaking to BS agar was the single most productive selective enrichment broth-agar combination for recovery of Salmonella in 5 of these food types. A study of the performance of these 5 agars used individually and in various combinations, showed that none of the 5 agars used individually nor any of the possible paired combinations of these agars could be used to satisfactorily detect Salmonella in the 9 food types. The use of all 5 agars was not necessary because one combination of 4 agars (BG, BS, HE, and XLD) recovered 100% of the Salmonella isolates, as compared with the number of Salmonella isolates recovered by the 5-agar combination, in each food category. This particular 4-agar combination, along with two 3-agar combinations (BG, BS, and XLD agars, and BS, HE, and XLD agars), were each able to recover more Salmonella isolates than the combination of BG, BS, and SS agars, the combination currently recommended by the AOAC. Finally, the relative costs of using these agars, singly and in various combinations, were determined.

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.

scholarly journals Detection of Burkholderia pseudomallei in Sputum using Selective Enrichment Broth and Ashdown’s Medium at Kampong Cham Provincial Hospital, Cambodia

F1000Research ◽  
2014 ◽  
Vol 3 ◽  
pp. 302 ◽  
Author(s):  
Somary Nhem ◽  
Joanne Letchford ◽  
Chea Meas ◽  
Sovanndeth Thann ◽  
James C. McLaughlin ◽  
...  
Keyword(s):  
Burkholderia Pseudomallei ◽  
Low Frequency ◽  
Culture Methods ◽  
Enrichment Broth ◽  
Selective Enrichment ◽  
Provincial Hospital ◽  
Testing Protocol ◽  
Standard Culture ◽  
Diagnostic Microbiology ◽  
Selective Enrichment Broth

Melioidosis infection, caused by Burkholderia pseudomallei, is increasingly reported in Cambodia. We hypothesized that implementation of an enhanced sputum testing protocol in a provincial hospital diagnostic microbiology laboratory would increase detection of B. pseudomallei. We tested 241 sputum specimens that were deemed acceptable for culture, comparing culture in selective enrichment broth followed by sub-culture on Ashdown’s medium to standard culture methods. Two specimens (0.8%) were positive for B. pseudomallei using the enhanced protocol whereas one specimen (0.4%) was positive using standard methods. These findings demonstrate that B. pseudomallei is rarely detected in sputum at this hospital. The low frequency of B. pseudomallei in sputum specimens precludes drawing any conclusions about the relative benefits of an enhanced sputum testing protocol at this site. Promoting clinician awareness of the infection and encouraging utilization of diagnostic microbiology services are likely to be important factors in facilitating identification of melioidosis.

Selective enrichment broth medium for isolation of Campylobacter jejuni.

1983 ◽  
Vol 17 (5) ◽  
pp. 853-855 ◽  
Author(s):  
W T Martin ◽  
C M Patton ◽  
G K Morris ◽  
M E Potter ◽  
N D Puhr
Keyword(s):  
Campylobacter Jejuni ◽  
Enrichment Broth ◽  
Selective Enrichment ◽  
Selective Enrichment Broth
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