'Evidence for waterborne origin of an extended mixed gastroenteritis outbreak in a town in Northern Greece, 2019'.

We investigated a large gastroenteritis outbreak that occurred in Northern Greece in 2019. A case was defined as anyone presenting with diarrhoea and/or vomiting from 24 January 2019 to 04 February 2019. We conducted a case-control study (CCS) using random selection of participants >16 years of age, residents of town X, who visited the health care centre between 25 and 28 January 2019. Moreover, we conducted a retrospective cohort study (CS) at the four elementary schools of the town. We collected clinical and water samples and the water supply system was inspected. In total, we recorded 638 cases (53% female; median age was 44 years (range 0-93)). Forty-eight cases and 52 controls participated in the CCS and 236 students in the CS. Both CCS and CS indicated tap water as the most likely source (OR 10, 95% CI 2.09-93.4, explaining 95.7% of cases; RR = 2.22, 95% CI 1.42-3.46, respectively). More than one pathogen was detected from stool samples of 6 of the 11 cases tested (norovirus, Campylobacter jejuni, Enterohemorrhagic E. coli (EHEC) and Enteropathogenic E. coli (EPEC)). Water samples, collected after ad-hoc chlorination, tested negative. Technical failures of the water tanks' status were identified. Our results suggested a waterborne outbreak. We recommended regular monitoring of the water supply system and immediate repair of technical failures.

Laboratory capacity of Greek hospitals for diagnosis of salmonellosis and surveillance systems' performance in the years of economic crisis, 2010-2016.

Austerity might have affected the capacity of public hospitals in Greece to diagnose salmonellosis (laboratory capacity) over the period 2010-2016, as well as the performance of the existing surveillance systems. The scope of this paper is to present data on laboratory capacity over these years, as well as the results of a two-source capture-recapture study (data from Mandatory Notification System and National Reference Laboratory System for Salmonella). The main findings were that: (a) laboratory capacity was high and steady besides the financial crisis, (b) the estimated number of laboratory-confirmed cases (n = 6017, 95% CI 5892-6142) resulted in an incidence rate (7.9 cases/100 000 population) almost twice than that reported by the two systems Mandatory Notification System (MNS); 4.1 and National Reference Laboratory System (NRLS); 4.5 cases/100 000 population, (c) underreporting was high for both systems (MNS; 47.5% and NRLS; 42.8%) and (d) differences by geographical region, size and type of hospital were identified. We suggest that (a) specific interventions are needed to increase completeness of the systems by type of hospital and geographical region, (b) record linkage can help in estimating the disease burden in a more valid way than each system separately and (c) a common electronic database in order to feed one system to the other could significantly increase completeness of both systems.

A severe gastroenteritis outbreak of Salmonella enterica serovar Enteritidis PT8, with PFGE profile XbaI.0024 and MLVA profile 2-9-7-3-2 following a christening reception, Greece, 2016.

In June 2016, a Salmonella enterica serovar Enteritidis outbreak (n = 56) occurred after a christening reception in Central Greece, mainly affecting previously healthy adults; one related death caused media attention. Patients suffered from profuse diarrhoea, fever and frequent vomiting episodes requiring prolonged hospitalisation and sick leave from work, with a 54% hospital admission rate. The majority of cases experienced serious illness within <12 h of attending the party. We investigated the outbreak to identify the source(s) of infection and contributing factors to the disease severity. From the retrospective cohort study, the cheesy penne pasta was the most likely vehicle of infection (relative risk 7·8; 95% confidence interval 3·6-16·8), explaining 79% of the cases. S. enterica ser. Enteritidis isolates were typed as phage-type PT8, pulsed-field gel electrophoresis type XbaI.0024, multiple locus variable-number tandem repeat analysis-type 2-9-7-3-2. The strain did not share the single-nucleotide polymorphism address of the concurrent European S. enterica ser. Enteritidis PT8 outbreak clusters. Following five consecutive years with no documented S. enterica ser. Enteritidis outbreaks in Greece, this outbreak, likely associated with a virulent strain, prompted actions towards the enhancement of the national Salmonella molecular surveillance and control programmes including the intensification of training of food handlers for preventing similar outbreaks in the future. Advanced molecular techniques were useful in distinguishing unrelated outbreak strains.

Resistant Shigella strains in refugees, August-October 2015, Greece.

Shigellosis is endemic in most developing countries and thus a known risk in refugees and internally displaced persons. In 2015, a massive influx of refugees into Greece, due to the political crisis in the Middle East, led to the development of appropriate conditions for outbreaks of communicable diseases as shigellosis. We present a cluster of 16 shigellosis cases in refugees, detected by the implementation of a syndromic notification system in one transit centre in Athens, between 20 August and 7 October 2015. Both Shigella flexneri (n = 8) and S. sonnei (n = 8) were identified, distributed in various serotypes. All tested isolates (n = 13) were multidrug resistant; seven were CTX-M-type extended-spectrum ß-lactamase producers. Our results indicate lack of a potential common source, although pulsed-field gel electrophoresis typing results revealed small clusters in isolates of the same serotype indicating possible limited person-to-person transmission without identifying secondary community cases related to the refugees. To prevent the spread of shigellosis, empirical antibiotic treatment as well as environmental hygiene measures were implemented. The detection of multi-drug resistance is important for determining the appropriate empirical antibiotic treatment for the more severe cases, while at the same time real-time typing is useful for epidemiological investigation and control measures.

Comparative study of all Salmonella enterica serovar Enteritidis strains isolated from food and food animals in Greece from 2008 to 2010 with clinical isolates.

The aim of the present work was to study the epidemiology of Salmonella enterica serovar Enteritidis (S. Enteritidis) in Greece, comparing all the food and food animal isolates during a 3-year period with clinical isolates. Submission of the generated data to the PulseNet Europe database was carried out in order to study the population structure of this particular serovar and indicate possible connections with European strains. One hundred and sixty-eight (168) S. Enteritidis strains of human, animal, and food origin, isolated during the period 2008-2010 in Greece, were studied. Strains were characterized by phenotypic (antibiotic resistance) and molecular [pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST)] methods. PFGE revealed 39 XbaI, 48 BlnI, and 80 XbaI-BlnI distinct pulsotypes, suggesting several clones circulating through the food chain and multiple sources of transmission. Submission to the PulseNet Europe database indicated that PFGE profile SENTXB.0001, the most common PFGE profile in Europe, was also predominant in Greece (33.3 %). MLST showed that all the strains studied shared the same sequence type (ST11), representing the most common ST in Europe. High rates of resistance to nalidixic acid were observed among human and poultry isolates (~25 %), indicating the potential fluoroquinolone treatment failure. Our data suggest that strains originating from multiple reservoirs circulated in Greece through the food chain during the study period. Predominant profiles in Greece were common to PulseNet Europe profiles, indicating similarities between the S. Enteritidis populations in Greece and Europe.

Performance of three small-scale wastewater treatment plants. A challenge for possible re use.

The study focused on the assessment of the performance of three WWTPs in Greece by the estimation of the microbiological and chemical quality of influent and effluent sewage. Physicochemical parameters were recorded (temperature, pH, COD, BOD, suspended solids, conductivity), and meteorological data were collected (air temperature, rain). Microbiological parameters were analyzed (Escherichia coli, total coliforms, bacteriophages, Salmonella, human adenoviruses, Candida, Pseudallescheria boydii, helminths, parasites Cryptosporidium ssp., and Giardia spp.). Statistically significant correlations among the various aforementioned parameters were investigated, in an attempt to propose appropriate processing performance indicators. Furthermore, the study aimed to assess current joint ministerial decision (JMD) on wastewater reuse, for irrigation purposes; to evaluate its practicability and its potential for public health protection. In the vast majority, outlet samples from all three studied WWTPs were not appropriate for irrigation reuse purposes based on BOD50 and suspended solids limit values, set by the current JMD, for both limited and unrestricted irrigation applications. Reductions for E. coli, total coliforms, and bacteriophages were found to range between 2-3, 1.5-2.5, and 2-4 log10 values, respectively. Salmonella spp. was detected in outlet sewage samples from Patra (PAT), Arachova (ARH), and Livadeia (LEV), at 23% (3/13), 33% (4/12), and 38% (5/13), respectively. Molds were detected at 92.3% (12/13), 100% (13/13), and 91.6% (11/12), respectively, while Candida was found at 85% (11/13), 67% (8/12), and 46% (6/13). A high prevalence of Pseudallescheria boydii, in outlet samples from all studied WWTPs is an important public health issue, which underlines the need for further studies on this emerging fungal pathogen in wastewater reuse applications. Pseudallescheria boydii was found at 85% (11/13), 67% (8/12), and 46% (6/13), respectively. Helminths were found in both inlet and outlet samples, of all studied WWTPs, at 100%. Human adenoviruses, were detected at high percentages in outlet samples at 76.9% (10/13), 92.3% (12/13), 84.6% (11/13), respectively, while no influence of UV irradiation was recorded on the entry and exit loads of human adenoviruses. No influence of meteorological parameters was found on the microbiological and chemical parameters, with the exception of a weak positive correlation between environmental temperature and bacteriophages. A moderate positive correlation was found between BOD and suspended solids, bacteriophages, and total coliforms, bacteriophages and E. coli, and bacteriophages and adenoviruses. A significant positive correlation was found between total coliforms and E. coli, COD and BOD, and suspended solids and COD. No correlations were proved between human pathogens and bacterial indicator parameters. Collectively, our findings underlined the unsuitability of the current JMD on wastewater reuse in Greece, or public health protection. The study is expected to support the development of a public health risk assessment model based on quantitative risk assessment on the use of treated wastewater for irrigation purposes in Greece.

Phenotypic and molecular characterization of multidrug-resistant Salmonella enterica serovar Hadar in Greece, from 2007 to 2010.

All 120 strains of Salmonella enterica serovar Hadar isolated during 2007-2010 in Greece were characterized by phenotypic and molecular methods. High rates of resistance to nalidixic acid (92%) and low levels of ciprofloxacin resistance (88%) were observed. Pulsenet-pulsed field gel electrophoresis profile SHADXB.0001 was predominant in Greece (58%) as in Europe but PT1, a rare phage type in Europe, was frequent in Greece (56%). The SHADXB.0001 and PT1 clone (38%) were found in humans, animals and food of animal origin with R-type ApSpTNxpCp being predominant (25%). The data indicate that this clone (possibly endemic) was circulating through the food chain in Greece during the study period.

Phenotypic and molecular characterisation of multiresistant monophasic Salmonella Typhimurium (1,4,[5],12:i:-) in Greece, 2006 to 2011.

Recently, multiresistant Salmonella enterica serovar1,4,[5],12:i:-, a monophasic variant of S. Typhimurium(1,4,[5],12:i:1,2) emerged, and is now among the most common serovars isolated from humans in many countries.In Greece, monophasic Typhimurium which was recorded for the first time in human isolates in 2007(0.3% of total isolates), increased sharply thereafter,and since 2009 is the third most frequent serovar. In the present study, 119 S. enterica 1,4,[5],12:i:- strains of human, animal and food origin, isolated during the period between 2006 and 2011, were examined.Strains verified as monophasic Typhimurium variants by polymerase chain reaction (PCR) (97 strains), were further characterised by phenotypic (antibiotic resistance and phage typing) and molecular (pulsed-field gel electrophoresis ­ PFGE) methods. The results indicate that multiple clones of multiresistant monophasic Typhimurium are circulating in Greece. The most frequently encountered clone in humans and pigs was that of phage type DT120, R-type ASSuTSpTm and PFGEprofile STYMXB.0010, while in poultry other clones were detected. The data indicate that pigs may be a reservoir of this clone in Greece.

Typing of Pseudomonas aeruginosa strains isolated from Greek water samples by three typing methods: serotyping, Random Amplified Polymorphic DNA (RAPD) and Pulsed Field Gel Electrophoresis (PFGE).

The study of various types of Pseudomonas aeruginosa strains isolated from water environments is of paramount importance from a public health point of view, due to their ubiquity and pathogenicity. Molecular (Random Amplified Polymorphic DNA and Pulsed Field Gel Electrophoresis) and phenotypical (serotyping) typing methods were applied to environmental P. aeruginosa strains. The typeability and discriminatory power of the methods were studied and compared. The two molecular methods managed to type a number of P. aeruginosa strains which were non-serotypeable due to their rough phenotypes. According to our results, the combination of phenotypic and genotypic methods increased the reliability of the results, yielding several different clones that seem to circulate in Greek water environments.

Equivalency testing of TTC Tergitol 7 agar (ISO 9308-1:2000) with five culture media for the detection of E. coli in water samples in Greece.

In this study ten laboratories in Greece compared the performance of reference method TTC Tergitol 7 Agar (with the additional test of beta-glucuronidase production) with five alternative methods, to detect E. coli in water, in line with European Water Directive recommendations. The samples were prepared by spiking drinking water with sewage effluent following a standard protocol. Chlorinated and non-chlorinated samples were used. The statistical analysis was based on the mean relative difference of confirmed counts and was performed in line with ISO 17994. The results showed that in total, three of the alternative methods (Chromocult Coliform agar, Membrane Lauryl Sulfate agar and Trypton Bilex-glucuronidase medium) were not different from TTC Tergitol 7 agar (TTC Tergitol 7 agar vs Chromocult Coliform agar, 294 samples, mean RD% 5.55; vs MLSA, 302 samples, mean RD% 1; vs TBX, 297 samples, mean RD% -2.78). The other two alternative methods (Membrane Faecal coliform medium and Colilert 18/ Quantitray) gave significantly higher counts than TTC Tergitol 7 agar (TTC Tergitol 7 agar vs MFc, 303 samples, mean RD% 8.81; vs Colilert-18/Quantitray, 76 samples, mean RD% 18.91). In other words, the alternative methods generated performance that was as reliable as, or even better than, the reference method. This study will help laboratories in Greece overcome culture and counting problems deriving from the EU reference method for E. coli counts in water samples.

Microbial risk assessment of dental unit water systems in general dental practice in Greece.

Water was sampled from source water, the 3-in-1 syringe and the air rotor water line of dental unit water systems (DUWS) in general dental practice in Attica, Greece. A section of the water line supplying the 3-in-1 syringe was cut for biofilm analysis. High total viable counts, Mycobacterium spp. and Pseudomonas aeruginosa were detected in the samples. Back siphonage was evidenced by the presence of blood and the recovery of oral anaerobes from the samples. Legionella pneumophila was recovered from 16.1% of biofilm samples. All tested disinfectants reduced the total viable counts of the DUWS to < 100 CFU/mL.

Attitudes of general dental practitioners in Europe to the microbial risk associated with dental unit water systems.

UNLABELLED: Dental Unit Water Systems (DUWS) are used in dental practices to provide water for cooling of dental equipment and irrigation of the oral cavity. However, they have been demonstrated to be contaminated with micro-organisms. There are currently no European Union (EU) Commission guidelines for the microbial quality of water discharged by DUWS. This study was part of an EU research programme to investigate the microbial contamination of DUWS in general dental practice (GDP) in the UK, Denmark, Germany, The Netherlands, Ireland, Greece and Spain. OBJECTIVE: To undertake a questionnaire survey on the type of DUWS in use and determine the attitude of GDPs to the risk of microbial infection from DUWS. MATERIALS AND METHODS: The questionnaire was written and translated into the language of each country before being posted to each participating dentist. Dentists were asked to complete the questionnaire survey and return it by post. RESULTS AND CONCLUSIONS: The major findings were that the majority of dentists did not clean, disinfect or determine the microbial load of their DUWS, and that dentists would welcome regular monitoring and advice on maintaining their DUWS; the introduction of guidelines; and recommendations on controlling the microbial load of DUWS.

The use of bacteriophages for monitoring the microbiological quality of sewage sludge.

The use of bacteriophages as potential indicators of faecal pollution has recently been studied. The correlation of the number of bacterial indicators and the presence of three groups of bacteriophages, namely somatic coliphages (SOMCPH), F-RNA specific phages (FRNAPH) and phages of Bacteroides fragilis (BFRPH), in raw and treated sludge is presented in this study. Raw and anaerobically digested sewage sludge samples from two wastewater treatment plants in Athens were collected on a monthly basis, over a 2-year period, and analyzed for total coliforms, E. coli, intestinal enterococci and the three groups of bacteriophages. A clear correlation between the number of bacterial indicators and the presence of bacteriophages was observed. E. coli concentrations of > or =10(3) cfus g(-1) and <10(3) cfus g(-1) comprise a threshold for the presence of FRNAPH and BFRPH, respectively. Likewise, intestinal enterococci concentrations of > or =10(4) cfus g(-1) and <10(3) cfus g(-1) comprise a threshold for the presence of FRNAPH and BFRPH, respectively. In the case of SOMCPH, it was not possible to define a threshold, since they were detected with the lowest observed indicator concentrations in all samples.

Comparison of the efficacies of disinfectants to control microbial contamination in dental unit water systems in general dental practices across the European Union.

Water delivered by dental unit water systems (DUWS) in general dental practices can harbor high numbers of bacteria, including opportunistic pathogens. Biofilms on tubing within DUWS provide a reservoir for microorganisms and should be controlled. This study compared disinfection products for their ability to meet the American Dental Association's guideline of <200 CFU x ml(-1) for DUWS water. Alpron, BioBlue, Dentosept, Oxygenal, Sanosil, Sterilex Ultra, and Ster4Spray were tested in DUWS (n = 134) in Denmark, Germany, Greece, Ireland, The Netherlands, Spain, and the United Kingdom. Weekly water samples were tested for total viable counts (TVCs) on yeast extract agar, and, where possible, the effects of products on established biofilm (TVCs) were measured. A 4- to 5-week baseline measurement period was followed by 6 to 8 weeks of disinfection (intermittent or continuous product application). DUWS water TVCs before disinfection ranged from 0 to 5.41 log CFU x ml(-1). Disinfectants achieved reductions in the median water TVC ranging from 0.69 (Ster4Spray) to 3.11 (Dentosept) log CFU x ml(-1), although occasional high values (up to 4.88 log CFU x ml(-1)) occurred with all products. Before treatment, 64% of all baseline samples exceeded American Dental Association guidelines, compared to only 17% following commencement of treatment; where tested, biofilm TVCs were reduced to below detectable levels. The antimicrobial efficacies of products varied (e.g., 91% of water samples from DUWS treated with Dentosept or Oxygenal met American Dental Association guidelines, compared to 60% of those treated with Ster4Spray). Overall, the continuously applied products performed better than those applied intermittently. The most effective products were Dentosept and Oxygenal, although Dentosept gave the most consistent and sustained antimicrobial effect over time.

Microbiological evaluation of dental unit water systems in general dental practice in Europe.

A range of opportunistic pathogens have been associated with dental unit water systems (DUWS), particularly in the biofilms that can line the tubing. This study therefore aimed to assess the microbiology of DUWS and biofilms in general dental practices across seven European countries, including the United Kingdom (UK), Ireland (IRL), Greece (GR), Spain (ES), Germany (D), Denmark (DK) and the Netherlands (NL). Water supplied by 51% of 237 dental unit water lines exceeded current American Dental Association recommendations of < or = 200 colony-forming units (CFU) ml(-1). Microbiological loading of the source waters was between 0 (Denmark, the Netherlands and Spain) and 4.67 (IRL) log CFU ml(-1); water line samples from the DUWS ranged from 1.52 (ES) to 2.79 (GR) log CFU ml(-1); and biofilm counts ranged from 1.49 (GR) to 3.22 (DK) log CFU.cm(-2). Opportunistic pathogens such as legionellae (DK and ES), including Legionella pneumophila SG1 (DK and GR), and Mycobacterium spp. (DK, NL, GR, D and ES) were recovered occasionally. Presumptive oral streptococci (ES and NL), oral anaerobes (GR), Candida spp. (UK, NL and ES) and blood (GR and IRL) were detected at relatively low frequencies, but their presence indicated a failure of the 3-in-1 antiretraction valve, leading to back siphonage of oral fluids into the water and biofilm phase. These findings confirm that a substantial proportion of DUWS have high levels of microbial contamination, irrespective of country, type of equipment and source water. The study emphasizes the need for effective mechanisms to reduce the microbial burden within DUWS, and highlights the risk of occupational exposure and cross-infection in general dental practice.

HbF production in beta thalassaemia heterozygotes for the IVS-II-1 G-->A beta(0)-globin mutation. Implication of the haplotype and the (G)gamma-158 C-->T mutation on the HbF level.

We studied the presence of the XmnI site and the beta-globin haplotype in 24 individuals, carriers of the IVSII-1 G-->A beta(0)-globin mutation, of whom fourteen had no detectable levels of HbF, while ten coming from 5 families, presented HbF levels ranging from 1.7 to 9% of the total Hb. Of these beta-thalassaemia heterozygotes with fetal hemoglobin, 6 were females and 4 were males with median HbF levels of 4.85% and 4% respectively, and an excess of (G)gamma chains (range (G)gamma/(A)gamma: 55/45-70/30). Of the group of carriers of beta-thalassaemia with HbF < 0.1, in all cases except one, IVSII-1 mutation was found associated with XmnI polymorphic site. Haplotype analysis in these individuals revealed that in 10 cases IVSII-1 was linked with haplotype IIIb, in 1 case with haplotype IIIa, and in 3 cases with haplotype IX. On the other hand, in the group of carriers with measurable levels of HbF, IVSII-1 was always associated with haplotype IIIa and the XmnI site was either in-homozygous or the heterozygous state in-cis or in-trans with the mutated beta-globin gene. In conclusion the results of the study of these families seem that XmnI site in-cis with the IVSII-1 does not induce HbF production when this beta(0)-thalassaemia mutation is associated with IIIb or IX haplotype. On the other hand the (G)gamma -158 C-->T mutation is associated with small amounts of HbF in IVSII-1 heterozygotes, when the beta-globin mutation is linked to haplotype IIIa.