Identification and evaluation of antimicrobial resistance of enterococci isolated from raw ewes’ and cows’ milk collected in western Sicily: a preliminary investigation

  • Marisa Palmeri Institute for Experimental Veterinary Medicine of Sicily A. Mirri, Palermo, Italy.
  • Isabella Mancuso Institute for Experimental Veterinary Medicine of Sicily A. Mirri, Palermo, Italy.
  • Raimondo Gaglio Department of Agricultural, Food and Forest Sciences, University of Palermo, Palermo, Italy.
  • Luigi Arcuri Local Health Unit, Palermo, Italy.
  • Santino Barreca Institute for Experimental Veterinary Medicine of Sicily A. Mirri, Palermo, Italy.
  • Pietro Barbaccia Department of Agricultural, Food and Forest Sciences, University of Palermo, Palermo, Italy.
  • Maria Luisa Scatassa | Institute for Experimental Veterinary Medicine of Sicily A. Mirri, Palermo, Italy.


The present work was carried out to investigate the Antimicrobial Resistance (AMR) of enterococci isolated from raw ewes’ and cows’ milk. The samples were collected from eighteen semi-extensive dairy sheep and cow farms throughout western Sicily. Plate counts, carried out on Rapid Enterococcus Agar commonly used to detect food enterococci, revealed a maximal enterococcal concentration of approximately 4.58 Log Colony Forming Unit (CFU)/mL. Colonies were isolated and differentiated based on genetic analysis by Randomly Amplified Polymorphic DNA (RAPD)-PCR. Thirty-eight different strains were identified. Analysis by a species-specific multiplex PCR assay grouped the strains into three Enterococcus species such as Enterococcus durans, Enterococcus faecalis and Enterococcus faecium. The 38 strains were also investigated for their antimicrobial resistance by a phenotypic approach. All 38 Enterococcus displayed resistance to at least one or more of the antimicrobials tested confirmed that the dairy enterococci could be a vector for the dissemination of antimicrobial resistance. This work showed that enterococci with AMR traits are commonly present in semiextensive dairy sheep and cow farms of western Sicily pointed out the relevance of informing dairy makers and veterinary regarding the antimicrobial use in order to mitigate problems of public health and veterinary medicine.



PlumX Metrics


Download data is not yet available.


Allington DR, Rivey MP, 2001. Quinupristin/dalfopristin: a therapeutic review. Clin Ther 23:24-44. DOI:

Çitak S, Yucel N, Orhan S, 2004. Antibiotic resistance and incidence of Enterococcus species in Turkish white cheese. Int J Dairy Technol 57:27-31. DOI:

Clinical and Laboratory Standards Institute (CLSI) 2017. Performance Standards for Antimicrobial Susceptibility Testing: 27th ed. CLSI Supplement M100. Wayne, PA: Clinical and Laboratory Standards Institute.

Chow JW, 2000. Aminoglycoside resistance in enterococci. Clin Infect Dis 31:586-89. DOI:

Cruciata M, Gaglio R, Scatassa ML, Sala G, Cardamone C, Palmeri M, Moschetti G, La Mantia T, Settanni L, 2018. Formation and characterization of early bacterial biofilms on different wood typologies applied in dairy production. Appl Environ Microbiol 84:e02107-17. DOI:

Cruciata M, Gaglio R, Todaro M, Settanni L, 2019. Ecology of Vastedda della valle del Belìce cheeses: A review and recent findings to stabilize the traditional production. Food Rev Int 35:90-103. DOI:

EFSA BIOHAZ Panel (EFSA Panel on Biological Hazards). Update of the list of QPS-recommended biological agents intentionally added to food or feed as notified to EFSA 4: Suitability of taxonomic units notified to EFSA until March 2016. EFSA J 2016; 14:4522. DOI:

Fernández L, Langa S, Martín V, Maldonado A, Jiménez E, Martín R, Rodríguez JM, 2012. The human milk microbiota: origin and potential roles in health and disease. Pharmacol Res 69:1-10. DOI:

Foulquié Moreno M, Sarantinopoulos P, Tsakalidou E, De Vuyst L, 2006. The role and application of enterococci in food and health. Int J Food Microbiol 106:1-24. DOI:

Franciosi E, Settanni L, Cologna N, Cavazza A, Poznanski E, 2011. Microbial analysis of raw cows’ milk used for cheese-making: influence of storage treatments on microbial composition and other technological traits. World J Microb Biot 27:171-80. DOI:

Franz CM, Holzapfel WH, Stiles ME, 1999. Enterococci at the crossroads of food safety? Int J Food Microbiol 47:1-24. DOI:

Gaglio R, Francesca N, Maniaci G, Corona O, Alfonzo A, Giosuè C, Di Noto AM, Cardamone C, Sardina MT, Portolano B, Alabiso M, 2016a. Valorization of indigenous dairy cattle breed through salami production. Meat Sci 114:58-68. DOI:

Gaglio R, Couto N, Marques C, Lopes MDFS, Moschetti G, Pomba C, Settanni L, 2016b. Evaluation of antimicrobial resistance and virulence of enterococci from equipment surfaces, raw materials, and traditional cheeses. Int J Food Microbiol 236:107-14. DOI:

Gaglio R, Francesca N, Di Gerlando R, Mahony J, De Martino S, Stucchi C, Moschetti G, Settanni L, 2017. Enteric bacteria of food ice and their survival in alcoholic beverages and soft drinks. Food Microbiol 67:17-22. DOI:

Gaglio R, Cruciata M, Scatassa ML, Tolone M, Mancuso I, Cardamone C, Corona O, Todaro M, Settanni L, 2019a. Influence of the early bacterial biofilms developed on vats made with seven wood types on PDO Vastedda della valle del Belìce cheese characteristics. Int J Food Microbiol 291:91-103 DOI:

Gaglio R., Todaro M., Scatassa ML, Franciosi E, Corona O, Mancuso I, Di Gerlando R, Cardamone C, Settanni L, 2019b. Transformation of raw ewes’ milk applying “Grana” type pressed cheese technology: Development of extra-hard “Gran Ovino” cheese. Int J Food Microbiol 307:108277. DOI:

Giraffa G, 2002. Enterococci from foods. FEMS Rev. Microbiol 26:163-71. DOI:

Guarcello R, De Angelis, M, Settanni L, Formiglio S, Gaglio R, Minervini F, Moschetti G, Gobbetti M, 2016a. Selection of amine-oxidizing dairy lactic acid bacteria and identification of the enzyme and gene involved in the decrease of biogenic amines. Appl Environ Microbiol 82:6870-80. DOI:

Guarcello R, Carpino S, Gaglio R, Pino A, Rapisarda T, Caggia C, Marino G, Randazzo CL, Settanni L, Todaro M, 2016b. A large factory-scale application of selected autochthonous lactic acid bacteria for PDO Pecorino Siciliano cheese production. Food Microbiol 59:66-75. DOI:

Guzman Prieto AM, van Schaik W, Rogers MR, Coque TM, Baquero F, Corander J, Willems RJ, 2016. Global emergence and dissemination of enterococci as nosocomial pathogens: attack of the clones? Front Microbiol 7:788. DOI:

Hammad AM, Hassan HA, Shimamoto T, 2015. Prevalence, antibiotic resistance and virulence of Enterococcus spp. in Egyptian fresh raw milk cheese. Food Control 50:815-20. DOI:

Jackson CR, Fedorka-Cray PJ, Barrett JB, 2004. Use of a genus- and species-specific multiplex PCR for identification of enterococci. J Clin Microbiol 42:3558–65. DOI:

Kang II-B, Kim DH, Chon JW, Seo KH, 2017. Effect of microbial control measures on farmstead cheesmaking and antimicrobial resistance of Staphylococcus aureus and Enterococcus spp. isolates. J Food Saf 38:e12432. DOI:

Licitra G, Ogier JC, Parayre S, Pediliggieri C, Carnemolla TM, Falentin H, Maden MN, Carpino S, Lortal S, 2007. Variability of bacterial biofilms of the “tina” wood vats used in the Ragusano cheese-making process. Appl Environ Microbiol 73:6980-87. DOI:

Macaluso G, Fiorenza G, Gaglio R, Mancuso I, Scatassa ML, 2016. In vitro evaluation of bacteriocin-like inhibitory substances produced by lactic acid bacteria isolated during traditional Sicilian cheese making. Ital J Food Saf 5:5503. DOI:

Mannu L, Paba A, Daga E, Comunian R, Zanetti S, Dupre` I, Sechi LA, 2003. Comparison of the incidence of virulence determinants and antibiotic resistance between Enterococcus faecium strains of dairy, animal and clinical origin. Int J Food Microbiol 88:291-304. DOI:

Russo N, Caggia C, Pino A, Coque TM, Arioli S, Randazzo CL, 2018. Enterococcus spp. In Ragusano PDO and Pecorino Siciliano cheese types: a snapshot of their antibiotic resistance distribution. Food Chem Toxicol 120: 277-86. DOI:

Scatassa ML, Gaglio R, Cardamone C, Macaluso G, Arcuri L, Todaro M, Mancuso I, 2017. Anti-Listeria activity of lactic acid bacteria in two traditional Sicilian cheeses. Ital J Food Saf 6:13-17. DOI:

Silvetti T, Morandi S, Brasca M, 2019. Does Enterococcus faecalis from traditional raw milk cheeses serve as reservoir of antibiotic resistance and pathogenic traits? Foodborne Pathog Dis 16:359-67. DOI:

Sitzia M, Bonanno A, Todaro M, Cannas A, Atzori AS, Francesconi AHD, Trabalza-Marinucci M, 2015. Feeding and management techniques to favour summer sheep milk and cheese production in the Mediterranean environment. Small Rumin Res 126:43-58. DOI:

Original Articles
Enterococcus; antimicrobial resistance; sheep milk; cow milk
  • Abstract views: 133

  • PDF: 80
  • HTML: 0
How to Cite
Palmeri M, Mancuso I, Gaglio R, Arcuri L, Barreca S, Barbaccia P, Scatassa ML. Identification and evaluation of antimicrobial resistance of enterococci isolated from raw ewes’ and cows’ milk collected in western Sicily: a preliminary investigation. Ital J Food Safety [Internet]. 2021Jan.19 [cited 2021Apr.15];9(4). Available from:

Most read articles by the same author(s)