Evaluation of the Impact of Varied Carvacrol Concentrations on Salmonella Recovery in Oregano and How Corn Oil Can Minimize the Effect of Carvacrol during Preenrichment.

Phenolic compounds, like carvacrol, in oregano interfere with the detection of foodborne pathogens such as Salmonella enterica. Carvacrol concentration varies based on plant cultivars and growth region. Six oregano cultivars were used to compare the impact of carvacrol concentration on Salmonella and to evaluate the effectiveness of corn oil to help increase Salmonella survival for detection. The results of Agilent 1200 series high-performance liquid chromatography analysis showed that carvacrol concentration in the six oregano cultivars ranged from 64 to 11,200 ppm. Oregano samples were artificially contaminated with S. enterica and were preenriched in Trypticase soy broth with or without 2% (v/v) corn oil. After 18 to 24 h at 37°C, aliquots were transferred to selective enrichment broths. Salmonella was recovered onto xylose lysine Tergitol 4 agar. Six Salmonella serovars were compared, and recovery varied based on carvacrol concentration and serovar. Samples with higher concentrations of carvacrol showed Salmonella recovery only when they were preenriched with corn oil. Based on metagenomic analysis, the microflora associated with the oregano also varied per cultivar. The results show that, as carvacrol levels increased, Salmonella survival decreased. However, the addition of corn oil to the preenrichment broth can minimize the antimicrobial effects of the phenolic compounds, thus allowing for increased detection of Salmonella from oregano cultivars.

Evaluation of corn oil as an additive in the pre-enrichment step to increase recovery of Salmonella enterica from oregano.

Phenolic compounds associated with essential oils of spices and herbs possess a variety of antioxidant and antimicrobial properties that interfere with Salmonella detection from fresh and dried products. Finding a compound to neutralize the effect of these antimicrobial compounds, while allowing Salmonella growth during pre-enrichment, is a crucial step in both traditional pathogen isolation and molecular detection from these foods. This study evaluated the effectiveness of corn oil as a component of the pre-enrichment broth to counteract antimicrobial compounds properties and increase the recovery of Salmonella from spices. Oregano samples artificially contaminated with Salmonella enterica were pre-enriched in modified Buffered Peptone Water (mBPW) supplemented with and without 2% (vol/vol) corn oil respectively. Samples were incubated overnight at 37 °C. The results showed that recovery of Salmonella from oregano samples was increased by ≥50% when pre-enriched with corn oil. Serovars were confirmed using a PCR serotyping method. In addition, shot-gun metagenomics analyses demonstrated bacterial diversity and the effect of corn oil on the relative prevalence of Salmonella in the oregano samples. Modifying pre-enrichment broths with corn oil improved the detection and isolation of Salmonella from oregano, and may provide an alternative method for pathogen detection in dried food matrices such as spices.

Are leaf glandular trichomes of oregano hospitable habitats for bacterial growth?

Phyllospheric bacteria were isolated from microsites around essential-oil-containing glands of two oregano (Origanum vulgare subsp. hirtum) lines. These bacteria, 20 isolates in total, were subjected to bioassays to examine their growth potential in the presence of essential oils at different concentrations. Although there were qualitative and quantitative differences in the essential oil composition between the two oregano lines, no differences were recorded in their antibacterial activity. In disk diffusion bioassays, four of the isolated strains could grow almost unrestrained in the presence of oregano oil, another five proved very sensitive, and the remaining 11 showed intermediate sensitivity. The strain least inhibited by oregano essential oil was further identified by complete16s rRNA gene sequencing as Pseudomonas putida. It was capable of forming biofilms even in the presence of oregano oil at high concentrations. Resistance of P. putida to oregano oil was further elaborated by microwell dilution bioassays, and its topology on oregano leaves was studied by electron microscopy. When inoculated on intact oregano plants, P. putida was able not only to colonize sites adjacent to essential oil-containing glands, but even to grow intracellularly. This is the first time that such prolific bacterial growth inside the glands has been visually observed. Results of this study further revealed that several bacteria can be established on oregano leaves, suggesting that these bacteria have attributes that allow them to tolerate or benefit from oregano secondary metabolites.

Arbuscular mycorrhiza alter the concentration of essential oils in oregano (Origanum sp., Lamiaceae).

The effect of root colonization by Glomus mosseae on the qualitative and quantitative pattern of essential oils (EO) was determined in three oregano genotypes (Origanum sp.). To exclude a simple P-mediated effect through mycorrhization the effect of P application to plants on the EO accumulation was also tested. In two genotypes the leaf biomass was increased through mycorrhization. Root colonization by the arbuscular mycorrhizal fungus (AMF) did not have any significant effect on the EO composition in oregano; however, in two genotypes the EO concentration significantly increased. As EO levels in P-treated plants were not enhanced, we conclude that the EO increase observed in mycorrhizal oregano plants is not due to an improved P status in mycorrhizal plants, but depends directly on the AMF-oregano plant association.

Fates of seeded Escherichia coli O157:H7 and Salmonella on selected fresh culinary herbs during refrigerated storage.

The fates of seeded Escherichia coli O157:H7 and Salmonella on selected fresh culinary herbs were evaluated at a refrigerated temperature (4 degrees C). Fresh herbs, including cilantro, oregano, basil, chive, parsley, and rosemary, were inoculated with six-strain mixtures of E. coli O157:H7 and Salmonella, and the microbial populations were monitored at 1, 5, 11, 16, 19, and 24 days. For both pathogens, a significant decrease in the population (P < 0.0001) occurred within the first 5 days of storage (< 0.8 log). Both pathogens remained the highest on cilantro and the lowest on rosemary (P < 0.0001). Storage time had a significant effect on the survival of E. coli O157:H7; populations declined as storage time progressed. Although storage of cilantro, basil, and chive was terminated after 19 days because of deteriorated quality, significant numbers of both pathogens were recovered from the remaining fresh herbs after 24 days of storage. The results showed that both bacteria were extremely persistent on all test herbs under the test conditions. The results also reinforce the concept that, once contaminated, bacterial pathogens can persist on fresh herbs throughout a normal distribution time.