M1UKStreptococcus pyogenes causing community-acquired pneumonia, pleural empyema and streptococcal toxic shock syndrome.

OBJECTIVES: Streptococcus pyogenes causes superficial infections but can also cause deep-seated infections and toxin-mediated diseases. In the present study, phylogenetic and in silico prediction analyses were performed on an antimicrobial resistant M1UKS. pyogenes strain causing severe clinical manifestations during the current surge of invasive group A Streptococcus (iGAS) disease. METHODS: A 40-year-old patient was admitted to the hospital with fever, chest pain and fatigue. Based on the clinical and laboratory findings, a diagnosis of sepsis with disseminated intravascular coagulation, community-acquired pneumonia, pleural empyema and streptococcal toxic shock syndrome was made. Microbial identification was performed by multiplex PCR and conventional culturing. Furthermore, antimicrobial susceptibility testing, whole genome sequencing, phylogenomic analysis and in silico prediction analysis of antimicrobial resistance genes and virulence factors were performed. RESULTS: S. pyogenes isolates were detected in pleural fluid and sputum of the patient. Both isolates belonged to the M1UK lineage of the emm1/ST28 clone, being closely related with an M1UK GAS strain from Australia. They exhibited resistance to erythromycin and clindamycin and susceptibility-increased exposure to levofloxacin and carried genes encoding for protein homologues of antibiotic efflux pumps. Moreover, several virulence factors, and a previously described single-nucleotide polymorphism in the 5' transcriptional leader sequence of the ssrA gene, which enhances expression of SpeA, were detected. CONCLUSIONS: The present antimicrobial-resistant M1UKS. pyogenes strain represents the first report of this emerging lineage associated with such manifestations of iGAS disease.

ICU-Associated Gram-Negative Bloodstream Infection: Risk Factors Affecting the Outcome Following the Emergence of Colistin-Resistant Isolates in a Regional Greek Hospital.

Intensive care unit patients may present infections by difficult-to-treat-resistant Gram-negative microorganisms. Colistin resurfaced as a last resort antibiotic for the treatment of multi-drug-resistant Gram-negative bacteria. However, colistin might not improve survival, particularly after the emergence of colistin-resistant isolates. We aimed to (1) examine the first Gram-negative-associated-bloodstream infection (GN-BSI) effect on 28-day mortality and (2) distinguish mortality risk factors. From 1 January 2018 to 31 December 2019, we retrospectively studied all adult patients admitted for more than 48 h in the critical care department of a regional Greek hospital, with prevalent difficult-to-treat Gram-negative pathogens. We examined the patient records for the first GN-BSI. The local laboratory used broth microdilution to evaluate bacterial susceptibility to colistin. Seventy-eight patients fulfilled the entry criteria: adult and first GN-BSI. They developed GN-BSI on day 10 (6-18), while the overall mortality was 26.9%. Thirty-two and 46 individuals comprised the respective colistin-resistant and colistin-sensitive groups. The admission Acute Physiology Assessment and Chronic Health Evaluation II score was associated with acquiring colistin-resistant GN-BSI in the multivariable logistic regression analysis (οdds ratio (CI), 1.11 (1.03-1.21)). Regarding mortality, the index day sequential organ failure assessment score was solely associated with the outcome (hazard-ratio (CI), 1.23 (1.03-1.48), Cox proportional hazard analysis). GN-BSI was often caused by colistin-resistant bacteria. Concerning our data, sepsis severity was the independent predictor of mortality regardless of the colistin-resistance phenotype or empirical colistin treatment.

Differences in sheep and goats milk microbiological profile between conventional and organic farming systems in Greece.

The aim of this study was to examine differences in the microbiological profile and antimicrobial resistance of bacteria isolated from milk from organic and conventional sheep and goat farms. Twenty-five organic and 25 conventional sheep and goat farms in the region of Thessaly, Greece participated in this study. A standardised detailed questionnaire was used to describe farming practices. A total of 50 samples were collected and analysed for total viable count (TVC), total coliform count (TCC) and somatic cell count (SCC), while Staphylococcus aureus and Escherichia coli were isolated using standard methods. Isolates were identified at species level by Api-test and Matrix-Assisted Laser Desorption/Ionisation-Time of Flight Mass Spectrometry (MALDI-TOF MS). Susceptibility to a panel of 20 for E. coli and 16 for S. aureus antimicrobials was determined by the agar dilution method. Pulsed Field Gel Electrophoresis (PFGE) was performed for S. aureus and E. coli isolates to determine predominant clones. Lower counts of TVC, TCC and SCC were identified in milk from the organic farms, possibly due to differences in the hygienic farming practices found on those farms. API-tests and MALDI-TOF MS showed no significant differences in the S. aureus and E. coli isolates. Overall, antimicrobial resistance rates were low, while a statistically higher percentage was estimated among strains originating from conventional farms in comparison with organic farms, possibly due to the restriction of antibiotic use in organic farming. PFGE revealed diversity among S. aureus and E. coli populations in both organic and conventional farms indicating circulation of 2-3 main clones changing slightly during their evolution. Consequently, there is evidence that milk from the organic farms presents a better microbiological profile when compared with milk from conventional farms.

Epidemiological Investigation of Legionella pneumophila Serogroup 2 to 14 Isolates from Water Samples by Amplified Fragment Length Polymorphism and Sequence-Based Typing and Detection of Virulence Traits.

The aim of this study is to explore the dispersion, clonality, and virulence of Legionella pneumophila serogroups 2 to 14 in the Greek environment. Eighty L. pneumophila serogroup 2 to 14 strains isolated from water distribution systems of hotels, hospitals, athletic venues, and ferries in Greece were tested by monoclonal antibodies (MAbs) for serogroup discrimination and molecularly by amplified fragment length polymorphism (AFLP) for genetic diversity. Fifty-six of 80 strains were also typed by the sequence-based typing (SBT) method. Αll strains were further analyzed for detection of two pathogenicity loci: Legionella vir homologue (lvh) and repeats in structural toxin (rtxA). Thirty-seven strains (46.2%) belonged to serogroup 6, 26 strains (32.5%) to serogroup 3, and 7 (8.8%) to other serogroups (4, 5, 8, and 10). Ten strains (12.5%) were nontypeable (NT) into the known serogroups. Thirty-nine different AFLP types were found among the 80 L. pneumophila serogroup 2 to 14 strains, and 24 different SBT types were found among the 56 strains tested. Among the 80 strains, the lvh locus was present in 75 (93.8%), the rtxA locus was found in 76 (95%), and both loci were found in 73 (91.3%) strains. This study showed that there is genetic variability of L. pneumophila serogroups 2 to 14 in the Greek environment as well as a high percentage of the pathogenicity loci. Ιntroducing an effective diagnostic test for L. pneumophila serogroups 2 to 14 in urine and promoting the examination of respiratory specimens from patients hospitalized for pneumonia in Greek hospitals are essential. IMPORTANCE: In this study, the dispersion, clonality, and virulence of environmental isolates of Legionella pneumophila serogroups 2 to 14 (Lp2-14) in Greece were investigated. Genetic variability of Lp2-14 in the Greek environment was identified together with the presence of the pathogenicity loci in a high percentage of the isolates. Despite the high prevalence of Lp2-14 in the Greek environment, no clinical cases were reported, which may be due to underdiagnosis of the disease. Almost all the legionellosis cases are diagnosed in Greece by using the urine antigen test, which is specific for Lp1. There is an urgent need to improve the clinical diagnosis of legionellosis by introducing an effective diagnostic test for Lp2-14 in urine and by promoting the PCR examination of respiratory specimens from patients with compatible clinical symptoms.

Considerations on the current universal vaccination policy against hepatitis A in Greece after recent outbreaks.

Greece is the only European Union member state that in 2008 included hepatitis A (HAV) vaccine in the routine national childhood immunization program (NCIP). Given that the resources allocated to public health have dramatically decreased since 2008 and that Greece is a low endemicity country for the disease, the benefit from universal vaccination has been questioned. The aim of this paper is to summarize the available epidemiological data of the disease for 1982-2013, and discuss the effects of universal vaccination on disease morbidity. Descriptive analysis, ARIMA modeling and time series intervention analysis were conducted using surveillance data of acute HAV. A decreasing trend of HAV notification rate over the years was identified (p<0.001). However, universal vaccination (~ 80% vaccine coverage of children) had no significant effect on the annual number of reported cases (p = 0.261) and has resulted to a progressive increase of the average age of infection in the general population. The mean age of cases before the inclusion of the vaccine to NCIP (24.1 years, SD = 1.5) was significantly lower than the mean age of cases after 2008 (31.7 years, SD = 2.1) (p<0.001). In the last decade, one third of all reported cases were Roma (a population accounting for 1.5% of the country's total population) and in 2013 three outbreaks with 16, 9 and 25 Roma cases respectively, were recorded, indicating the decreased effectiveness of the current immunization strategy in this group. Data suggest that universal vaccination may need to be re-considered. Probably a more cost effective approach would be to implement a program that will include: a) vaccination of high risk groups, b) universal vaccination of Roma children and improving conditions at Roma camps, c) education of the population and travel advice, and d) enhancement of the control measures to increase safety of shellfish and other foods.

Association of treatment for bacterial meningitis with the development of sequelae.

BACKGROUND: Bacterial meningitis continues to be a serious, often disabling infectious disease. The aim of this study was to assess the possibility that treatment influences the development of sequelae in childhood bacterial meningitis. METHODS: Two thousand four hundred and seventy-seven patients aged 1 month to 14 years with acute bacterial meningitis over a 32-year period were enrolled in the study. Data were collected prospectively from the Meningitis Registry of a tertiary university teaching hospital in Athens, Greece. Treatment was evaluated through univariate and multivariate analysis with regard to sequelae: seizure disorder, severe hearing loss, ventriculitis, and hydrocephalus. RESULTS: According to the multinomial logistic regression analysis, there was evidence that penicillin, an all-time classic antibiotic, had a protective effect on the occurrence of ventriculitis (odds ratio (OR) 0.17, 95% confidence interval (CI) 0.05-0.60), while patients treated with chloramphenicol had an elevated risk of ventriculitis (OR 17.77 95% CI 4.36-72.41) and seizure disorder (OR 4.72, 95% CI 1.12-19.96). Cephalosporins were related to an increased risk of hydrocephalus (OR 5.24, 95% CI 1.05-26.29) and ventriculitis (OR 5.72, 95% CI 1.27-25.76). The use of trimethoprim/sulfamethoxazole increased the probability of seizure disorder (OR 3.26, 95% CI 1.08-9.84) and ventriculitis (OR 8.60, 95% CI 2.97-24.91). Hydrocortisone was associated with a rise in hydrocephalus (OR 5.44, 95% CI 1.23-23.45), while a protective effect of dexamethasone (OR 0.82, 95% CI 0.18-3.79) was not statistically significant. CONCLUSIONS: Current study findings suggest that the type of antimicrobial treatment for childhood bacterial meningitis may influence in either a positive or a negative way the development of neurological sequelae.

Pertussis Epidemiology in Greece and Emerging Risk Groups during the Vaccination Era (1980-2008).

To study the epidemiology of pertussis in Greece and epidemiologic changes throughout a period of twenty-nine years, we conducted a retrospective analysis of available data of pertussis cases for the past twenty-nine years (1980-2008) and a prospective analysis of hospitalized pertussis cases from a children's hospital in Athens for eight years (2001-2008). From 1980 through 2008, the incidence of pertussis in Greece declined from 11.2 to 0.05 cases per 100,000. Epidemic cycles occurring every 3 to 5 years were observed. Since pertussis circulation cannot be fully controlled by present immunization programs, efforts should be made to vaccinate infants at the recommended age, early diagnose, treatment as well as contact tracing of pertussis cases. Control of pertussis in social susceptible populations is necessary. A national program with adolescent and adult booster could decrease the circulation of B. pertussis. Despite an overall decrease for pertussis cases, pertussis is still a present and future challenge of public health service in Greece.

Elevated bathing-associated disease risks despite certified water quality: a cohort study.

Bacteriological water quality criteria have been recommended to ensure bathers' health. However, this risk-assessment approach is based mainly on routine measurements of fecal pollution indicator bacteria in seawater, and may not be adequate to protect bathers effectively. The aim of this study was to assess the risks of symptoms related to infectious diseases among bathers after exposure to seawater which was of excellent quality according to EU guidelines. This study is a cohort study recruiting bathers and non-bathers. Water samples were collected for estimating bacterial indicators. Univariable and multivariable analysis was performed to compare the risks of developing symptoms/diseases between bathers and non-bathers. A total of 3805 bathers and 572 non-bathers were included in the study. Water analysis results demonstrated excellent quality of bathing water. Significantly increased risks of symptoms related to gastrointestinal infections (OR = 3.60, 95% CI 1.28-10.13), respiratory infections (OR = 1.92, 95% CI 1.00-3.67), eye infections (OR = 2.43, 95% CI 1.27-4.63) and ear infections (OR = 17.21, 95% CI 2.42-122.34) were observed among bathers compared with non-bathers. Increased rates of medical consultation and medication use were also observed among bathers. There was evidence that bathers experienced increased morbidity compared with non-bathers though the bathing waters met bacteriological water quality criteria. These results suggest that risk assessments of recreational seawaters should not only focus on bacteriological water quality criteria.

Decreasing trend of Legionella isolation in a long-term microbial monitoring program in Greek hospitals.

Legionnaires' disease is a known cause of severe pneumonia in a nosocomial setting. This study examined Legionella isolation in Greek hospitals. Water samples and swabs of showerheads from 41 hospitals were collected over a four-year period (2004-2007) from hot and cold water systems and cooling towers in Greece. In total, 1058 samples were examined for the presence of Legionella. From the hot water samples examined, 166 out of 607 (27.3%) were positive for Legionella, including 111 (18.3%) positive for Legionella pneumophila sg1 samples. The temperature of hot water samples less than 55°C was statistically significant, associated with Legionella spp. isolation (RR 4.01, 95%CI 2.33-6.92). Ten out of 17 (58.8%) hospital cooling towers required remedial actions due to Legionella colonization. Reemergence of Legionella spp. colonization was evident in more than half of the hospitals where frequent monitoring and appropriate risk assessment plans were absent or lacking. Hospitals that were monitored continuously over the study period presented a decreasing trend of Legionella colonization. Environmental risk assessment together with Legionella isolation should be enforced systematically in hospitals.

Bather density as a predominant factor for health effects related to recreational bathing: results from the Greek bathers cohort study.

Our aim was to examine the health effects on bathers from exposure to recreational seawater on three beaches with different microbiology quality and bather density through a cohort study. An initial questionnaire and a 10 day follow up were used. Univariate and backward logistic regression analyses were used to asses the risk of developing symptoms or diseases among the bathers. In total, 3805 bathers were included in the study and 149 samples were collected from the three beaches. Despite all the beaches being characterized of high quality the levels of bacterial indicators differed among them. Health effects among bathers were not associated with bacterial indicators. A statistically significant increased risk for symptoms related to respiratory illness, gastroenteritis, medical consultation and use of medication was observed among bathers at the higher bather density beaches. Beach bather numbers should always be taken into account when considering a risk assessment approach.