Different microbial groups of the microbiome of fresh produce can have diverse effects on human health. This study was aimed at identifying some microbial communities of fresh produce by analyzing 105 samples of imported fresh fruits and vegetables originated from different countries in the world including local samples (Oman) for aerobic plate count and the counts of Enterobacteriaceae, Enterococcus, and Staphylococcus aureus. The isolated bacteria were identified by molecular (PCR) and biochemical methods (VITEK 2). Enterobacteriaceae occurred in 60% of fruits and 91% of vegetables. Enterococcus was isolated from 20% of fruits and 42% of vegetables. E. coli and S. aureus were isolated from 22% and 7% of vegetables, respectively. Ninety-seven bacteria comprising 21 species were similarly identified by VITEK 2 and PCR to species level. E. coli, Klebsiella pneumoniae, Enterococcus casseliflavus, and Enterobacter cloacae were the most abundant species; many are known as opportunistic pathogens which may raise concern to improve the microbial quality of fresh produce. Phylogenetic trees showed no relationship between clustering of the isolates based on the 16S rRNA gene and the original countries of fresh produce. Intercountry passage of opportunistic pathogens in fresh produce cannot be ruled out, which requires better management.

Enterobacteria may gain antibiotic resistance and be potent pathogens wherever they are present, including in fresh fruits and vegetables. This study tested the antibiotic resistance of enterobacteria isolated from 13 types of local and imported fresh fruits and vegetables (n = 105), using the standard Kirby-Bauer disk diffusion method. Phenotypic and genotypic characterizations of AmpC β-lactamases were determined in cefoxitin-resistant isolates. Ten percent of the enterobacteria tested (n = 88) were pansusceptible, 74% were resistant to at least one antibiotic, and 16% were multidrug resistant. Enterobacteria isolates showed the highest antibiotic resistance against ampicillin (66%), cephalothin (57%), amoxicillin–clavulanic acid (33%), cefoxitin (31%), tetracycline (9%), nalidixic acid (7%), trimethoprim (6%), and kanamycin (5%). Three isolates showed intermediate resistance to the clinically important antibiotic imipenem. Escherichia coli isolated from lettuce exhibited multidrug resistance against five antibiotics. Fifteen isolates were confirmed to have AmpC β-lactamase, using the inhibitor-based test and the antagonism test; the latter test confirmed that the enzyme was an inducible type. Four types of ampC β-lactamase genes (CIT, EBC, FOX, and MOX) were detected in eight isolates: four Enterobacter cloacae isolates and one isolate each of Citrobacter freundii, Enterobacter asburiae, Enterobacter hormaechei, and Enterobacter ludwigii. It was concluded that fresh fruits and vegetables might play a role as a source or vehicle for transferring antibiotic-resistant bacteria that might spread to other countries through exportation. The clinically significant AmpC β-lactamase was rarely documented in the literature on bacteria isolated from fruits and vegetables, and to our knowledge, this is the first report on the detection of an inducible type in such commodities.

The contamination of fresh produce with antibiotic-resistant bacteria is of particular concern as they are often eaten raw and can be a source for foodborne diseases. Tetracyclines have been largely used in humans, animals and plants which might have accelerated microbial resistance to them. Enterococci and Escherichia coli can be used as indicators to monitor contamination of the fresh produce with tetracycline-resistant bacteria. The investigation related to this issue is very scarce in Oman. This study aimed at identifying tetracycline-resistant enterococci and E. coli in fresh produce at the market place. Thirty-one enterococci and ten E. coli were isolated from local (Oman) and imported fruits and vegetables (N= 105). Using the standard Kirby-Bauer disc diffusion method, resistance to tetracycline was found in 6 (19 %) enterococci, isolated from cucumber, lettuce and radish, and 5 (50 %) E. coli, obtained from cabbage, lettuce and radish. Genetic analysis revealed the presence of tetracycline resistance genes, tet(A) and tet(K), in E. coli and tet(K), tet(L) and tet(M) in enterococci, including Enterococcus sulfureus, Enterococcus mundtii, Enterococcus casseli avus and Enterococcus faecalis. The integron integrase IntI 1 gene, which is known to facilitate the dissemination of antibiotic resistance genes among bacteria, was detected in 2 isolates of E. coli. These results demonstrated the capability of fresh produce to act as a potential source for disseminating tetracycline or possibly other antibiotic-resistant bacteria through the food chain. Thus, control strategies are needed to reduce exposure of the public to such microorganisms.

Fresh produce bacteria may have phenotypic and/or genotypic antimicrobial resistance traits that may lead to various consequences on the environment and human health. This study evaluated the susceptibility of fresh produce bacteria (banana, cabbage, capsicum, carrots, cucumber, dates, lettuce, mango, papaya, pomegranate, radish, tomato and watermelon) to chlorhexidine and the antibiotic resistance of enterococci. Eighty-eight Enterobacteriaceae bacteria and 31 enterococci were screened for their susceptibility to chlorhexidine using the broth microdilution method. Susceptibility of enterococci to various antibiotics was determined using agar dilution, colorimetric, and Kirby-Bauer disc diffusion methods. Enterococci were more susceptible to chlorhexidine than Enterobacteriaceae indicated by chlorhexidine minimum inhibitory concentration (MIC) of 1 to 8 µg/mL for the former and 1 to 64 µg/mL for the latter. The IntI 1, qacEΔ1, qacE and qacG genes were distributed weakly in three, two, two, and three Enterobacteriaceae isolates, respectively. Enterococci had resistance to chloramphenicol (3%), tetracycline (19%), erythromycin (68%), ciprofloxacin (55%), and vancomycin (10%) while 19% of them were multi-drug resistant. In conclusion, this research detected a low to moderate level of antibiotic resistance in enterococci. Some Enterobacteriaceae bacteria had reduced chlorhexidine MICs that were not 10x less than the recommended concentration (100–200 µg/mL) in food production areas which might challenge the success of the disinfection processes or have clinical implications if the involved bacteria are pathogens. The prevalence of antimicrobial-resistant bacteria in fresh produce should be monitored in the future.

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