Proceeding of Veterinary and Animal Science Days 2017, 6th- 8th June, Milan, Italy 

HAF © 2013 

Vol. IV, No. 1s   ISSN: 2283-3927 

   

 

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Keywords 

Staphylococcus epidermidis, 

Implant-related infections, 

Orthopedic infections, Biofilm, 

Genome sequencing 

 

CORRESPONDING AUTHOR 

Marta Bottagisio 

marta.bottagisio@unimi.it 

 

JOURNAL HOME PAGE 

riviste.unimi.it/index.php/haf 

Phenotypic and genomic identification of 

Staphylococcus epidermidis GOI1153754-03-14 

isolated from an infected orthopedic prosthesis 

Marta Bottagisio1,2*, Alessio Soggiu1, Arianna B. Lovati2,  

Marco Toscano3, Cristian Piras1, Carlo L. Romanò4, Luigi Bonizzi1,  

Paola Roncada5, Lorenzo Drago3,6 

 

1University of Milan, Department of Veterinary Medicine, Italy 

2IRCCS Galeazzi Orthopedic Institute Cell and Tissue Engineering Laboratory, Milan, Italy  

3University of Milan, Department of Biomedical Science for Health, Italy 

4IRCCS Galeazzi Orthopedic Institute Center for Reconstructive Surgery of Osteoarticular Infections 
Milan, Italy  

5Istituto Sperimentale Italiano Lazzaro Spallanzani, Rivolta D'Adda, Cremona, Italy 

6IRCCS Galeazzi Orthopedic Institute Laboratory of Clinical Chemistry and Microbiology, Milan, Italy 

 

 Abstract 

Staphylococcus epidermidis GOI1153754-03-14 is able to colonize orthopedic implants and to cause 

septic non-unions, as validated in a recent in vivo study (Lovati et al., 2016). To the mechanisms leading 

to the biofilm formation on metallic implants, we carried out the phenotypic and genotypic 

characterization of the clinical isolate S. epidermidis GOI1153754-03-14.  

The antimicrobial susceptibility and minimum inhibitory concentration (MIC) of the strain were 

evaluated through the Vitek2 System (Biomerieux), as well as its ability to form biofilm in vitro through 

a spectrophotometric assay (Stepanovich et al., 2000). 

The genomic DNA was extracted by Bacterial Genomic DNA Isolation Kit (Norgen Biotek Corp.). 

Libraries were prepared with the ThruPLEX DNA-seq (Rubicon Genomics) and then sequenced on the 

Illumina MiSeq platform through the MiSeq Reagent Kit v3 (600-cycles) to produce 300 bp paired-end 

reads (Illumina Inc.). Reads were quality-trimmed and gene annotated thanks to the RAST software 

(Aziz et al., 2008). 

The antimicrobial susceptibility along with the MIC values are reported in Table 1. The outputs resulted 

in 51 contigs (Average = 50,720.6 Mb) with 396X fold average coverage. The total genome is 2,586,753 

bp long with a GC content of 31.84% and an N50 value of 7 bp. The whole genome is composed by 

2,467 protein-encoding genes and 64 RNAs (55 tRNAs and 9 rRNAs). The entire genome sequence has 

been deposited in the European Nucleotide Archive (ENA) under the accession no. FWCG01000000 

(Bottagisio et al., 2017).  

The genotypic and phenotypic characterization of the S. epidermidis GOI1153754-03-14 will enable a 

better comprehension of the mechanisms involved in the biofilm formation on orthopedic implants, 

paving the way for innovative preventative and therapeutic strategies. Moreover, the sequence of 

this clinical strain is mandatory to develop dedicated proteomics analysis in order to highlight 

functional mechanism of biofilm formation. 
 

Acknowledgments 

This study was supported by the Italian Ministry of Health (#RC 2016, L4083) 

http://creativecommons.org/licenses/by-nc-nd/3.0/deed.en


Proceeding of Veterinary and Animal Science Days 2017, 6th- 8th June, Milan, Italy 

HAF © 2013 

Vol. IV, No. 1s   ISSN: 2283-3927 

 

 

 

 

 

 

 

 

 

 

 

Table 1: Antimicrobial susceptibility and MIC values 

Antibiotic Antimicrobial susceptibility MIC 

Benzylpenicillin Resistant ≥ 0.5 

Oxacillin Resistant ≥ 4 

Levofloxacin Resistant  4 

Rifampicin Resistant  ≥ 4 

Gentamicin Susceptible  ≤ 0.5 

Erythromycin Susceptible ≤ 0.25 

Clindamycin Susceptible ≤ 0.12 

Linezolid Susceptible  1 

Daptomycin Susceptible 0.25 

Teicoplanin Susceptible 4 

Vancomycin Susceptible ≤ 0.5 

Tetracycline Susceptible ≤ 1 

Tigecycline Susceptible ≤ 0.12 

Fusidic acid Susceptible ≤ 0.5 

Trimethoprim/ Sulfamethoxazole Susceptible ≤ 10 

 

 

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http://creativecommons.org/licenses/by-nc-nd/3.0/deed.en