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Applied and Environmental Microbiology, June 2006, p. 4028-4035, Vol. 72, No. 6
0099-2240/06/$08.00+0     doi:10.1128/AEM.00034-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Ecology of Antibiotic Resistance Genes: Characterization of Enterococci from Houseflies Collected in Food Settings{dagger}

Lilia Macovei and Ludek Zurek*

Department of Entomology, Kansas State University, Manhattan, Kansas 66506

Received 5 January 2006/ Accepted 29 March 2006

In this project, enterococci from the digestive tracts of 260 houseflies (Musca domestica L.) collected from five restaurants were characterized. Houseflies frequently (97% of the flies were positive) carried enterococci (mean, 3.1 x 103 CFU/fly). Using multiplex PCR, 205 of 355 randomly selected enterococcal isolates were identified and characterized. The majority of these isolates were Enterococcus faecalis (88.2%); in addition, 6.8% were E. faecium, and 4.9% were E. casseliflavus. E. faecalis isolates were phenotypically resistant to tetracycline (66.3%), erythromycin (23.8%), streptomycin (11.6%), ciprofloxacin (9.9%), and kanamycin (8.3%). Tetracycline resistance in E. faecalis was encoded by tet(M) (65.8%), tet(O) (1.7%), and tet(W) (0.8%). The majority (78.3%) of the erythromycin-resistant E. faecalis isolates carried erm(B). The conjugative transposon Tn916 and members of the Tn916/Tn1545 family were detected in 30.2% and 34.6% of the identified isolates, respectively. E. faecalis carried virulence genes, including a gelatinase gene (gelE; 70.7%), an aggregation substance gene (asa1; 33.2%), an enterococcus surface protein gene (esp; 8.8%), and a cytolysin gene (cylA; 8.8%). Phenotypic assays showed that 91.4% of the isolates with the gelE gene were gelatinolytic and that 46.7% of the isolates with the asa1 gene aggregated. All isolates with the cylA gene were hemolytic on human blood. This study showed that houseflies in food-handling and -serving facilities carry antibiotic-resistant and potentially virulent enterococci that have the capacity for horizontal transfer of antibiotic resistance genes to other bacteria.

* Corresponding author. Mailing address: Department of Entomology, Kansas State University, Manhattan, KS 66506. Phone: (785) 532-4731. Fax: (785) 532-6232. E-mail: lzurek{at}ksu.edu.

{dagger} Contribution no. 06-171-J from the Kansas Agricultural Experimental Station.

Applied and Environmental Microbiology, June 2006, p. 4028-4035, Vol. 72, No. 6
0099-2240/06/$08.00+0     doi:10.1128/AEM.00034-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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