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

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.

Level of Detection (LOD50) of Campylobacter Is Strongly Dependent on Strain, Enrichment Broth, and Food Matrix

“…Since the levels of this pathogen in food are generally low, and Campylobacter may be sublethally damaged in food matrices, an enrichment procedure is advised for the detection of this microorganism (Rogol et al, 1985 ; Kim et al, 2009 ). Several studies have been carried out to examine Campylobacter detection protocols showing differences in detection rates depending on the type of medium and antibiotic use (Ugarte-Ruiz et al, 2012 ; Jo et al, 2017 ). Within the International Organization for Standardization (ISO), teams of international experts drafted standards, and also a horizontal method for the detection of Campylobacter spp.…”

Estudio de los caracteres epidemiológicos y control experimental de "Campylobacter jejuni" en el entorno de la producción de pollos para carne = Study of the epidemiological characteristics and experimental control of "Campylobacter jejuni" in the area of poultry for meat production

“…La microbiota presente en el intestino de los pollos es muy elevada y diversa (Lu et al, 2003) y, al intentar aislar Campylobacter del contenido intestinal, los contaminantes con frecuencia exhiben un crecimiento excesivo en las placas, ya que los protocolos de detección actuales no son lo suficientemente selectivos como para inhibir microorganismos inespecíficos (Jo et al, 2017). Hasta el momento, no se ha documentado un medio o método estándar universalmente aceptado para la recuperación de…”

“…El uso de un caldo de enriquecimiento en la detección de C. jejuni es necesario para facilitar el crecimiento inicial de los microorganismos presentes en una muestra, pudiendo además, suprimir las bacterias competidoras que puedan existir en la muestra (Jo et al, 2017). Sin embargo, el enriquecimiento inadecuado en el caso de las muestras más contaminadas, como las heces, puede provocar niveles más escasos de crecimiento de Campylobacter, mientras se permite el aumento de la cantidad de contaminantes .…”

Rapid and reliable detection of foodborne pathogens : Exploring Campylobacter ecology in enrichments to improve detection

“…to restore the selectivity of media by adjusting the antibiotic cocktail Jo et al, 2017;Kim et al, 2016; in order to suppress the growth of ESBL-producers but unfortunately, these antibiotics sometimes also suppress the growth of campylobacters which can led to false-negative detection outcomes . The duration of the lag phase (λ), maximum specific growth rate (µmax) and maximum cell concentration reached in stationary phase (asymptote) can be estimated using Chapter 1 32 different mathematical models.…”