Spa diversity and genetic characterization of t127 methicillin-resistant Staphylococcus aureus in a tertiary Greek hospital

Introduction: Methicillin-resistant Staphylococcus aureus (MRSA) causes severe community and hospital acquired infections. Identification of staphylococcal cassette chromosome mec (SCCmec), multilocus-sequence typing, and sequencing of S. aureus protein A (spa) gene are used for MRSA typing. The aim was to investigate the spa types of MRSA isolates in a tertiary hospital in Greece and analyse the whole genome sequences of two t127 MRSA isolates. Methods: Totally, 39 MRSA isolates collected from July 2019 to June 2020 in "Georgios Gennimatas" General Hospital of Thessaloniki, Greece, were included in the study. Identification and antimicrobial susceptibility testing were performed using VITEK II automated system, and spa typing was performed. A minimum spanning tree was used to display the spa type frequencies and the genetic distances among them. Two t127-MRSA isolates (IM-MRSA and PD-MRSA) were selected for WGS. Results: Six isolates (15.4%) were resistant to mupirocin, 18 (46.2%) to fusidic acid, three (7.7%) to vancomycin and two (5.1%) to teicoplanin. Twenty-two different spa types were detected, with t002, t003, and t422 being the most frequent (5/39, 12.8% each), followed by t1994 (4/39, 10.3%). The isolates presented high genetic diversity and, taking into account the time between hospital admission and sampling, intrahospital spread did not occur. Even the two t127 isolates were assigned to different sequence types, ST9-XII-t127 and ST1-IVa-t127. Plasmids and genes conferring antimicrobial resistance and virulence were also identified. Conclusions: Various spa types were identified and together with the information about the time between hospital admission and sampling supports polyclonal MRSA spread in the hospital excluding a nosocomial infection. WGS provides a more detailed analysis distinguishing even the isolates belonging to the same spa type

Vascular Endothelial Growth Factor Levels in Dobrava/Belgrade Virus Infections

The levels of vascular endothelial growth factor-A (VEGF) were estimated in 102 serum samples from 63 hospitalized Greek patients with hemorrhagic fever with renal syndrome (HFRS) caused by Dobrava/Belgrade virus. Significantly higher VEGF levels were seen in the severe when compared with non-severe cases (mean values 851.96 pg/mL and 326.75 pg/mL, respectively; p = 0.003), while a significant difference was observed among groups based on the day after the onset of illness. In both severe and non-severe cases, VEGF peaked in the second week of illness; however, elevation of VEGF in the severe cases started later and remained high until convalescence, suggesting that the role of VEGF was associated with repair of vascular damage rather than with increased permeability

Immune response in patients with hemorrhagic fever with renal syndrome and in patients with Crimean-Congo hemorrhagic fever

Two viral hemorrhagic fevers have been reported in Greece, hemorrhagic fever with renal syndrome (HFRS) caused by hantaviruses (in Greece mainly by Dobrava-Belgrade virus, DOBV) and Crimean-Congo hemorrhagic fever (CCHF) caused by CCHF virus (CCHFV). Even though the pathogenesis of these diseases is not fully understood, the activation of immune response system and the secretion of cytokines, known as “cytokine storm”, is thought to play a major role. Aim of the present thesis was to investigate the immune response in patients with HFRS and in patients with CCHF, focused on the evaluation of the cytokine patterns in relation with the clinical course and/or the outcome of the disease.Materials and methods: The study included 54 blood samples collected 5-70 days (mean 13.5 days) after onset of the disease from 30 HFRS patients (26 males) aged 17-70 years (median age 35.5 years), and 29 blood samples collected 1-9 days (mean 4.9 days) after onset of the disease from 29 CCHF patients (12 males) aged 4-67 years (median age 44 years). One HFRS case was fatal and 17 were considered as severe, while two CCHF cases were fatal and 8 were considered as severe. The samples were divided into groups according to the clinical course, the outcome of the disease and the days collected post symptoms' onset (day of illness). Serum samples from 16 apparently healthy individuals (9 males) aged 18-65 years (median age 45 years) were included in the study as control group. IgM and IgG antibodies against hantaviruses and CCHFV were detected using indirect immunofluorescent assay (IFA) and ELISA, while viral RNA was detected using RT-nested PCR and real time RT-PCR. The serum levels of 27 cytokines [interleukin (IL)-1b, IL-1ra, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12, IL-13, IL-15, IL-17, eotaxin, FGF-b, G-CSF, GM-CSF, IFN-γ, IP-10, MCP-1, MIP-1a, MIP-1b, PDGF-bb, RANTES, TNF-α and VEGF] were estimated using the BioPlexTM Suspension Array System (Bio-Rad Laboratories, CA).Results: IgM and IgG antibodies against hantaviruses were detected in all HFRS patients; hantavirus RNA was detected by RT-nested PCR in 22/29 (75.9%) samples which had been taken up to 16th day of illness. Sequencing of the PCR products showed that in all cases the causative agent was DOBV. During the first two weeks of illness, and compared to the control group, significantly elevated IL-1ra, IL-6, IL-8, IL-9, IL-10, GM-CSF, ΙΡ-10, ΜΙΡ-1b, PDGF, TNF-α and VEGF levels were observed in the severe cases, while in the non-severe cases, IL-13 and TNF-α levels were significantly elevated (p<0.05). In the two samples from the fatal case it was seen that the levels of IL-1b, IL-4, IL-7 IL-13, IL-17, MIP-1a and PDGF were decreased, while the levels of IL-6, IL-8, IL-10, IP-10 and MIP-1b were increased (p<0.05). A strong immune response was seen during the first week of illness, especially in severe cases, while the response in non-severe cases was weaker and delayed. During the 3rd week of illness or later and compared to the control group, serum levels of IL-9, IL-10, ΜΙΡ-1b, PDGF and VEGF remained elevated in severe cases, while levels of IL-6, IL-8, IL-10, IP-10 and MIP-1a remained elevated in the two samples from the fatal case (up to the death). In contrast in non-severe cases, the serum levels of most cytokines were similar to those of the control group. In all groups, IP-10 was increased and RANTES was decreased (p<0.05). Univariate logistic regression analysis showed that high serum levels of IL-8, IL-10 and VEGF were associated with disease severity, while multivariate regression analysis revealed VEGF as the most independent factor.Among the 29 CCHF patients, IgM antibodies were detected in 14 (48.3%), while IgG antibodies were not detected. Real-time RT-PCR was positive in all samples, while RT-nested PCR was positive in 23/29 (79.3%) samples. Compared to the control group, significantly elevated IL-6, IL-8, IL-9, IL-10, IL-13, GM-CSF, IFN-γ και IP-10 levels were found in severe cases (p<0,05), while in non-severe cases, increased levels were seen in IL-1ra, IL-6, IL-8, IL-10, IP-10, MCP-1 and VEGF (p<0,05). Levels of IL-1b, IL-6, IL-8, IL-9, IL-10, IL-15, eotaxin, ΙΡ-10, MCP-1 and TNF-α were increased in the fatal cases (p<0.05). In all groups, IL-6, IL-8, IL-10 and IP-10 were increased, with the highest levels being observed in the fatal cases (p<0.05). Among survivors, IP-10 and MCP-1 were higher in severe than in non-severe cases (p<0.05). Significantly higher levels of IL-1b, IL-6, IL-8, IL-9, IL-10, IL-15, ΙΡ-10, MCP-1 and TNF-α were observed in fatal than in non-fatal cases (p<0.05). The viral load was positively correlated with IL-10, IP-10, and MCP-1 levels, and negatively correlated with the ratio IL- 12/IL-10 (p<0.05). Multivariate logistic regression analysis showed that viral load and IP-10 were associated with disease severity and fatal outcome, while multivariate analysis revealed IP-10 as the most independent factor.Comparing the cytokine levels between HFRS and CCHF, several similarities and differences were noticed. No significant differences were found among fatal cases, while among survivors, mean levels of IL-8, IL-9, IL-10 and eotaxin were higher in HFRS patients (p<0.05). Compared to the control group, mean level of RANTES was decreased in both HFRS and CCHF patients (p<0.05). Similarly, in fatal HFRS and CCHF cases IP-10 was increased and IL-4 was decreased (p<0.05). Levels of IL-10 were increased only in fatal HFRS cases (p<0.05), while in fatal CCHF cases, MCP-1 and TNF-α were significantly increased and IL-12 was decreased (p<0.05). Mean levels of IL-6 and IL-8 were increased in HFRS cases and in fatal CCHF cases (p<0.05). VEGF was increased only in HFRS survivors (p<0.05). Conclusions: The present study showed that the immune response in HFRS and CCHF patients is time dependent and affects and is affected by the course and the outcome of the disease. Immune response in severe cases is stronger than in non-severe cases. VEGF and IP-10 can serve as possible biomarkers of disease severity in HFRS and in CCHF, respectively. The similarities of the immune response in HFRS and in CCHF suggest common pathophysiology of viral hemorrhagic fevers, while the observed differences are likely due to variations of interactions between viruses and the target host cells. The results of the study could be the basis of future research aiming to treatment and vaccine design.Οι ιογενείς αιμορραγικοί πυρετοί που ενδημούν στην Ελλάδα είναι ο αιμορραγικός πυρετός με νεφρική προσβολή (hemorrhagic fever with renal syndrome, HFRS) που οφείλεται σε χανταϊούς, και κυρίως στον ιό Dobrava-Belgrade (Dobrava-Belgrade virus, DOBV) και ο αιμορραγικός πυρετός Κριμαίας-Κονγκό (Crimean-Congo hemorrhagic fever, CCHF) που οφείλεται στον ομώνυμο ιό (CCHF virus, CCHFV). Οι παθογενετικοί μηχανισμοί των αιμορραγικών πυρετών δεν είναι επαρκώς γνωστοί, ωστόσο η ενεργοποίηση της ανοσιακής απάντησης και η υπερέκκριση κυτταροκινών, γνωστή ως "καταιγίδα κυτταροκινών" φαίνεται να παίζουν σημαντικό ρόλο. Σκοπός της παρούσας διατριβής ήταν η μελέτη της ανοσιακής απάντησης σε ασθενείς με HFRS και σε ασθενείς με CCHF, κυρίως όσον αφορά την εκτίμηση και την αξιολόγηση των επιπέδων των κυτταροκινών και την συσχέτιση αυτών με το στάδιο, την έκβαση και τη βαρύτητα της νόσου. Υλικό και μέθοδοι: Στη μελέτη περιλήφθηκαν 54 δείγματα αίματος που ελήφθησαν 5-70 μέρες (μέση ημέρα νόσου 13,5) μετά την έναρξη των συμπτωμάτων από 30 ασθενείς με HFRS (26 άνδρες) ηλικίας 17-70 ετών (διάμεση ηλικία 35,5 έτη), καθώς και 29 δείγματα αίματος που ελήφθησαν 1-9 ημέρες μετά την έναρξη των συμπτωμάτων (μέση ημέρα νόσου 4,9) από 29 ασθενείς με CCHF (12 άνδρες) ηλικίας 4-67 ετών (διάμεση ηλικία 44 έτη). Ένα περιστατικό HFRS ήταν θανατηφόρο, και άλλα 17 περιστατικά θεωρήθηκαν σοβαρά. Δύο περιστατικά CCHF ήταν θανατηφόρα, και άλλα 8 θεωρήθηκαν σοβαρά περιστατικά. Tα δείγματα των ασθενών διαχωρίστηκαν σε ομάδες με βάση την έκβαση, τη βαρύτητα και τον αριθμό των ημερών που μεσολάβησαν από την έναρξη των συμπτωμάτων μέχρι τη λήψη του δείγματος. Την ομάδα ελέγχου αποτέλεσαν 16 δείγματα από 16 υγιή άτομα (9 άνδρες) ηλικίας 18-65 ετών (διάμεση ηλικία 45 έτη).Τα δείγματα των ασθενών εξετάστηκαν για την ύπαρξη IgM και IgG αντισωμάτων έναντι των χανταϊών και του CCHFV με μεθόδους ELISA και έμμεσου ανοσοφθορισμού, ενώ η ανίχνευση του ιικού RNA έγινε με RT-nested PCR και real time RT-PCR. Τα επίπεδα 27 κυτταροκινών [ιντερλευκίνη (IL)-1b, IL-1ra, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12, IL-13, IL-15, IL-17, εοταξίνη, FGFb, G-CSF, GM-CSF, IFN-γ, IP-10, MCP-1, MIP-1a, MIP-1b, PDGF-bb, RANTES, TNF-α και VEGF] μετρήθηκαν ταυτόχρονα με τη χρήση της τεχνολογίας BioPlexTM Suspension Array System (Bio-Rad Laboratories, CA).Αποτελέσματα: Σε όλους τους ασθενείς με HFRS ανιχνεύτηκαν IgM και IgG αντισώματα έναντι των χανταϊών. Με RT-nested PCR ανιχνεύθηκε RNA χανταϊού, και συγκεκριμένα του DOBV, σε 22/29 (75,9%) δείγματα ασθενών που είχαν ληφθεί μέχρι την 16η ημέρα της νόσου. Κατά τις πρώτες δύο εβδομάδες της νόσου, και συγκριτικά με την ομάδα ελέγχου, τα σοβαρά περιστατικά παρουσίασαν αυξημένα επίπεδα των παρακάτω κυτταροκινών: IL-1ra, IL-6, IL-8, IL-9, IL-10, GM-CSF, ΙΡ-10, ΜΙΡ-1b, PDGF, TNF-α και VEGF (p<0,05). Τα ήπια περιστατικά παρουσίασαν αυξημένα επίπεδα των IL-13 και TNF-α (p<0,05). Στα δύο δείγματα από το θανατηφόρο περιστατικό HFRS παρατηρήθηκαν χαμηλά επίπεδα των IL-1b, IL-4, IL-7 IL-13, IL-17, MIP-1a και PDGF, ενώ τα επίπεδα των IL-6, IL-8, IL-10, IP-10 και MIP-1b ήταν υψηλά (p<0,05). Ισχυρή ανοσιακή απάντηση παρατηρήθηκε κατά την 1η εβδομάδα της νόσου, ιδιαίτερα στα σοβαρά περιστατικά, ενώ στα ήπια ήταν ηπιότερη και εμφανιζόταν αργότερα. Μετά το πέρας της 2ης εβδομάδας νόσου, και συγκριτικά με την ομάδα ελέγχου, τα σοβαρά περιστατικά διατήρησαν υψηλά τα επίπεδα των IL-9, IL-10, ΜΙΡ-1b, PDGF και VEGF, ενώ στα δύο δείγματα από το θανατηφόρο περιστατικό διατηρήθηκαν αυξημένα τα επίπεδα των IL-6, IL-8, IL-10, IP-10 και MIP-1a (p<0,05). Αντίθετα, η πλειοψηφία των κυτταροκινών στα ήπια περιστατικά ήταν σε επίπεδα παρόμοια με αυτά της ομάδας ελέγχου. Σε όλες τις ομάδες ασθενών τα επίπεδα της IP-10 ήταν αυξημένα και της RANTES μειωμένα (p<0,05). Η μονοπαραγοντική ανάλυση έδειξε ότι τα υψηλά επίπεδα των IL-8, IL-10 και VEGF παρουσίασαν σημαντική συσχέτιση με την εμφάνιση σοβαρής νόσου, ενώ η πολυπαραγοντική ανάλυση ανέδειξε τον VEGF ως τον πλέον ισχυρό παράγοντα. Μεταξύ των 29 ασθενών με CCHF, IgM αντισώματα έναντι του ιού ανιχνεύθηκαν σε 14 (48,3%). Σε κανέναν δεν ανιχνεύθηκαν αντισώματα IgG. Η Real-time RT-PCR ήταν θετική σε όλα τα δείγματα, ενώ η RT-nested PCR ήταν θετική σε 23/29 (79,3%) δείγματα. Συγκριτικά με την ομάδα ελέγχου τα σοβαρά περιστατικά παρουσίασαν αυξημένα επίπεδα των παρακάτω κυτταροκινών: IL-6, IL-8, IL-9, IL-10, IL-13, GM-CSF, IFN-γ και IP-10 (p<0,05). Τα ήπια περιστατικά παρουσίασαν αυξημένα επίπεδα των IL-1ra, IL-6, IL-8, IL-10, IP-10, MCP-1 και VEGF (p<0,05). Στα θανατηφόρα περιστατικά τα επίπεδα των IL-1b, IL-6, IL-8, IL-9, IL-10, IL-15, εοταξίνης, ΙΡ-10, MCP-1 και TNF-α ήταν αυξημένα (p<0,05). Τα επίπεδα των IL-6, IL-8, IL-10 και IP-10 ήταν αυξημένα και στις τρεις ομάδες σε σύγκριση με την ομάδα ελέγχου, με υψηλότερα επίπεδα στα θανατηφόρα περιστατικά (p<0,05). Μεταξύ των επιζώντων, τα επίπεδα των IP-10 και MCP-1 ήταν υψηλότερα στα σοβαρά περιστατικά συγκριτικά με τα ήπια. Τα επίπεδα των IL-1b, IL-6, IL-8, IL-9, IL-10, IL-15, ΙΡ-10, MCP-1 και TNF-α ήταν υψηλότερα στα θανατηφόρα περιστατικά συγκριτικά με τα μη θανατηφόρα (p<0,05). Το ιικό φορτίο παρουσίασε θετική συσχέτιση με τα επίπεδα των IL-10, ΙΡ-10 και MCP-1 και αρνητική συσχέτιση με τον λόγο IL-12/IL-10. Η μονοπαραγοντική ανάλυση έδειξε ότι το ιικό φορτίο και η ΙΡ-10 παρουσίασαν σημαντική συσχέτιση με την εμφάνιση σοβαρής νόσου, ενώ η πολυπαραγοντική ανάλυση ανέδειξε την ΙΡ-10 ως τον πλέον ισχυρό παράγοντα.Συγκρίνοντας τα επίπεδα των κυτταροκινών στους δύο ιογενείς αιμορραγικούς πυρετούς, στα μεν θανατηφόρα περιστατικά δεν παρατηρήθηκε σημαντική διαφορά, μεταξύ όμως των επιζώντων, οι μέσες τιμές των IL-8, IL-9, IL-10 και εοταξίνης ήταν σημαντικά υψηλότερες στους ασθενείς με HFRS (p<0,05). Συγκριτικά με την ομάδα ελέγχου, η RANTES ήταν ελαττωμένη σε όλους τους ασθενείς (p<0,05). Στα θανατηφόρα περιστατικά HFRS και CCHF η IP-10 ήταν αυξημένη και η IL-4 ελαττωμένη (p<0,05). Τα επίπεδα της IL-10 ήταν αυξημένα μόνο στα θανατηφόρα περιστατικά HFRS, ενώ μόνο στα θανατηφόρα περιστατικά CCHF, τα επίπεδα των MCP-1 και TNF-α ήταν αυξημένα (p<0,05). Η IL-6 και η IL-8 ήταν αυξημένη σε όλους τους ασθενείς με HFRS και στα θανατηφόρα περιστατικά CCHF (p<0,05). Ο VEGF ήταν αυξημένος μόνο στους επιζήσαντες του HFRS. Συμπεράσματα: Η παρούσα μελέτη έδειξε ότι η ανοσιακή απάντηση των ασθενών με HFRS και CCHF εξαρτάται σημαντικά από την χρονική φάση της νόσου, και επηρεάζεται, αλλά και επηρεάζει την κλινική πορεία και την έκβαση της νόσου. Η ανοσολογική απάντηση είναι εντονότερη στα σοβαρά περιστατικά HFRS και CCHF σε σύγκριση με τα ήπια περιστατικά. Γενικά, ο VEGF και η IP-10 είναι δυνατόν να θεωρηθούν ως προγνωστικοί δείκτες βαρύτητας της νόσου στον HFRS και τον CCHF, αντίστοιχα. Οι ομοιότητες της ανοσιακής απάντησης στον HFRS και τον CCHF είναι αποτέλεσμα της κοινής παθοφυσιολογίας των ιογενών αιμορραγικών πυρετών, ενώ οι διαφορές πιθανώς οφείλονται σε παραλλαγές των αλληλεπιδράσεων μεταξύ των ιών και των κυττάρων-στόχων του ξενιστή. Τα αποτελέσματα της μελέτης μπορούν να αποτελέσουν βάση για μελλοντικές έρευνες με στόχο τον σχεδιασμό θεραπευτικών σχημάτων και εμβολίων

Carbapenem-Resistant <i>Klebsiella pneumoniae</i>: Virulence Factors, Molecular Epidemiology and Latest Updates in Treatment Options

Klebsiella pneumoniae is a Gram-negative opportunistic pathogen responsible for a variety of community and hospital infections. Infections caused by carbapenem-resistant K. pneumoniae (CRKP) constitute a major threat for public health and are strongly associated with high rates of mortality, especially in immunocompromised and critically ill patients. Adhesive fimbriae, capsule, lipopolysaccharide (LPS), and siderophores or iron carriers constitute the main virulence factors which contribute to the pathogenicity of K. pneumoniae. Colistin and tigecycline constitute some of the last resorts for the treatment of CRKP infections. Carbapenemase production, especially K. pneumoniae carbapenemase (KPC) and metallo-β-lactamase (MBL), constitutes the basic molecular mechanism of CRKP emergence. Knowledge of the mechanism of CRKP appearance is crucial, as it can determine the selection of the most suitable antimicrobial agent among those most recently launched. Plazomicin, eravacycline, cefiderocol, temocillin, ceftolozane–tazobactam, imipenem–cilastatin/relebactam, meropenem–vaborbactam, ceftazidime–avibactam and aztreonam–avibactam constitute potent alternatives for treating CRKP infections. The aim of the current review is to highlight the virulence factors and molecular pathogenesis of CRKP and provide recent updates on the molecular epidemiology and antimicrobial treatment options

Carbapenem-Resistant Klebsiella pneumoniae: Virulence Factors, Molecular Epidemiology and Latest Updates in Treatment Options

Klebsiella pneumoniae is a Gram-negative opportunistic pathogen responsible for a variety of community and hospital infections. Infections caused by carbapenem-resistant K. pneumoniae (CRKP) constitute a major threat for public health and are strongly associated with high rates of mortality, especially in immunocompromised and critically ill patients. Adhesive fimbriae, capsule, lipopolysaccharide (LPS), and siderophores or iron carriers constitute the main virulence factors which contribute to the pathogenicity of K. pneumoniae. Colistin and tigecycline constitute some of the last resorts for the treatment of CRKP infections. Carbapenemase production, especially K. pneumoniae carbapenemase (KPC) and metallo-&beta;-lactamase (MBL), constitutes the basic molecular mechanism of CRKP emergence. Knowledge of the mechanism of CRKP appearance is crucial, as it can determine the selection of the most suitable antimicrobial agent among those most recently launched. Plazomicin, eravacycline, cefiderocol, temocillin, ceftolozane&ndash;tazobactam, imipenem&ndash;cilastatin/relebactam, meropenem&ndash;vaborbactam, ceftazidime&ndash;avibactam and aztreonam&ndash;avibactam constitute potent alternatives for treating CRKP infections. The aim of the current review is to highlight the virulence factors and molecular pathogenesis of CRKP and provide recent updates on the molecular epidemiology and antimicrobial treatment options

Crimean-Congo Hemorrhagic Fever: Tick-Host-Virus Interactions

Crimean-Congo hemorrhagic fever virus (CCHFV) is transmitted to humans by bite of infected ticks or by direct contact with blood or tissues of viremic patients or animals. It causes to humans a severe disease with fatality up to 30%. The current knowledge about the vector-host-CCHFV interactions is very limited due to the high-level containment required for CCHFV studies. Among ticks, Hyalomma spp. are considered the most competent virus vectors. CCHFV evades the tick immune response, and following its replication in the lining of the tick's midgut, it is disseminated by the hemolymph in the salivary glands and reproductive organs. The introduction of salivary gland secretions into the host cells is the major route via which CCHFV enters the host. Following an initial amplification at the site of inoculation, the virus is spread to the target organs. Apoptosis is induced via both intrinsic and extrinsic pathways. Genetic factors and immune status of the host may affect the release of cytokines which play a major role in disease progression and outcome. It is expected that the use of new technology of metabolomics, transcriptomics and proteomics will lead to improved understanding of CCHFV-host interactions and identify potential targets for blocking the CCHFV transmission

Genetic characterization of two methicillin-resistant Staphylococcus aureus spa type t127 strains isolated from workers in the dairy production chain in Greece

Methicillin-resistant Staphylococcus aureus (MRSA) constitutes a constant threat for the public health. Aim of the present study was to analyse the whole genome sequences of two MRSA strains belonging to Staphylococcus protein A (spa) type t127 isolated from humans working in two distantly located dairy production farms in Greece. MRSA strains were isolated from the nasal cavity of a food handler in a milk industry in Epirus, northwestern Greece (E-MRSA), and a person working in a cattle farm in Thrace, northeastern Greece (T-MRSA). Whole genome sequences taken using next generation sequencing were analysed for resistance and virulence genes applying various bioinformatic tools. Both isolates were assigned to ST1-IVa-t127 type, and they were transferring genes conferring resistance to tetracycline, beta-lactams, and aminoglycosides; T-MRSA was carrying additional genes leading to macrolide, lincosamide and streptogramin B (MLSB) resistance. Both isolates were carrying three plasmid replicon types, reps, rep7 and rep16, while T-MRSA harboured also rep10 and rep15. E-MRSA carried scn and sak genes which were absent from T-MRSA. In conclusion, the genetic characterization of two unrelated ST1-IVa-t127 MRSA strains isolated from humans in close contact with livestock in Greece can be used as basis for further epidemiological and evolutionary studies

Crimean-Congo Hemorrhagic Fever: CXCL10 Correlates With the Viral Load

Crimean-Congo hemorrhagic fever (CCHF) is a human disease with high fatality rate. Although its pathogenesis is not elucidated yet, it is considered that cytokines play a significant role in the progression and outcome of the disease. Serum CXCL10 levels were estimated in 35 patients with acute CCHF and were correlated with the viral load, and various demographic and clinical parameters. The mean CXCL10 concentration in the patients' group was higher compared to the respective value in the control group (4421.74pg/ml vs. 28.47pg/ml, P<0.05). A strong positive correlation between CXCL10 and viral load was seen (r(s)=0.57, P<0.001), while the outcome of the disease was related with the viral load (r(s)=0.47, P=0.004) and the presence of hemorrhagic manifestations (P<0.001). The study provides an insight into the strong correlation between CXCL10 and viral load in acute CCHF cases suggesting that it plays an important role in CCHF pathogenesis. J. Med. Virol. 87:899-903, 2015. (c) 2015 Wiley Periodicals, Inc