Molecular detection of multiple viral targets in untreated urban sewage from Greece. Kokkinos PA, Ziros PG, Mpalasopoulou A, Galanis A, Vantarakis A. Environmental Microbiology Unit, Department of Public Health, School of Medicine, University of Patras, Rion, GR 26504, Greece. Abstract Urban sewage virological analysis may produce important information about the strains that cause clinical and […]

Molecular detection of multiple viral targets in untreated urban sewage from Greece.
Kokkinos PA, Ziros PG, Mpalasopoulou A, Galanis A, Vantarakis A.

Environmental Microbiology Unit, Department of Public Health, School of Medicine, University of Patras, Rion, GR 26504, Greece.

Abstract
Urban sewage virological analysis may produce important information about the strains that cause clinical and subclinical infections in the population, thus supporting epidemiological studies.
In the present study, a twenty one-month survey (November 2007 to July 2009) was conducted in order to evaluate the presence of human adenoviruses (hAdV), hepatitis A viruses (HAV), hepatitis E viruses (HEV), Noroviruses (NoV), and human Polyomaviruses (hPyV) in untreated sewage samples collected from the inlet of Patras’ municipal biological wastewater treatment plant, located in southwestern Greece. Nucleic acid amplification techniques were applied for viral nucleic acid detection. Positive samples were confirmed by sequencing and comparative phylogenetic analysis was performed on the isolated viral strains.
In total, viruses were detected in 87.5% (42/48) of sewage samples. AdVs, PyVs, HAV, and NoVs were detected in 45.8% (22/48), 68.8% (33/48), 8.3% (4/48), and 6.3% (3/48) of the samples collected from the plant’s inlet, while HEV was not detected at all. Adenovirus types 8 (Ad8), 40 (Ad40) and 41 (Ad41) were recognized, while JC and BK polyomaviruses were recorded. Noroviruses were identified as GII.4. HAV was typed as genotype IA.
Our study demonstrates the advantages of environmental surveillance as a tool to elucidate the molecular epidemiology of community circulating viruses. We underline the need of environmental surveillance programs in countries such as Greece with inadequate and problematic epidemiological surveillance system and no environmental surveillance system currently in action.

Analysis of seawater microbiological quality data in Greece from 1997 to 2006: association of risk factors with bacterial indicators. Papastergiou P, Mouchtouri V, Karanika M, Kostara E, Kolokythopoulou F, Mpitsolas N, Papaioannou A, Hadjichristodoulou C. Department of Hygiene and Epidemiology, School of Medicine, University of Thessaly, Larissa 41221, Greece. Abstract This study deals with the […]

Department of Hygiene and Epidemiology, School of Medicine, University of Thessaly, Larissa 41221, Greece.

Abstract

This study deals with the examination of quality of seawater bathing areas in Greece over a 10-year period and identifies risk factors for high bacteria indicator organism concentrations. Qualitative descriptive analysis was applied and the microbiological test results of 231,205 water samples were associated with pollution markers and other parameters. Measurements of Escherichia coli (99.6%) and enterococci (100%) were found to be in accordance with the mandatory value guidelines set by the new European Directive. An increasing trend for the yearly mean value of faecal streptococci was noted. Using logistic regression analysis, phenolic smell (OR = 2.10, CI = 2.04-2.16), rainfall the day before sampling (OR = 1.67, CI = 1.64-1.74), high seas (OR = 1.42, CI = 1.39-1.46) and rainfall on the day of sampling (OR = 1.27, CI = 1.20-1.33) were positively independently associated with high levels of bacterial indicators (total coliforms, faecal coliforms, faecal streptococci and E. coli). The highest risk, absolute risk value 42.8% (RR = 3.17, CI = 2.97-3.38), was measured when previous day rainfall, phenolic smell and high seas were simultaneously recorded. Such parameters should be further investigated as predetermining factors for the assessment of beach bathing water quality, providing a timely indication of water risk assessment.

“Molecular characterization of hepatitis A virus isolates from environmental and clinical samples in Greece”, Petros Kokkinos, Panos Ziros, Sevasti Filippidou, Ioannis Mpampounakis, Apostolos Vantarakis Background: Hepatitis A virus (HAV) strains detected in environmental and clinical samples were analysed to characterize the genotypes of HAV circulating in Greece. Fifty (50) sewage samples were collected from Patras […]

“Molecular characterization of hepatitis A virus isolates from environmental and clinical samples in Greece”,
Petros Kokkinos, Panos Ziros, Sevasti Filippidou, Ioannis Mpampounakis, Apostolos Vantarakis

Background: Hepatitis A virus (HAV) strains detected in environmental and clinical samples were analysed to characterize the genotypes of HAV circulating in Greece. Fifty (50) sewage samples were collected from Patras (South-Western Greece) and Alexandroupolis (North-Eastern Greece) from 2007 until 2009, accordingly. The clinical samples derived from an HAV outbreak involved populations from three neighbouring prefectures of North-Eastern Greece (Xanthi, Rodopi, and Evros). HAV particles were detected by nested RT-PCR, using a previously validated set of primers to amplify a 290-bp fragment encompassing the 5’-NTR. Positive HAV samples were confirmed by sequencing of the PCR product. To determine the relatedness between the different isolated sequences, a phylogenetic tree was constructed.
Results: Results showed a 100% prevalence of genotype I, and particularly subgenotype IA. The analyzed HAV strains were closely related between them with the percentage of nucleotide identity ranging between 96% and 100%.
Conclusions: The study revealed the major prevalence of circulating strains of IA genotype in Greece and underlined the usefulness of molecular methods for the detection and typing of viruses in both environmental and clinical samples. The present study is, to our knowledge, the first in Greece to depict the simultaneous molecular characterization of HAV strains isolated from both clinical and environmental samples.

“Molecular typing of enteroviruses, adenoviruses and hepatitis A viruses in raw (untreated) and treated sewage water of a wastewater treatment plant in Greece” P. Kokkinos · S. Filippidou ·K.Karlou· A.Vantarakis Environmental Microbiology Unit, Department of Public Health, Medical School, University of Patras, Greece The sewage treatment plant effluents may contain infectious human viruses representing a […]

“Molecular typing of enteroviruses, adenoviruses and hepatitis A viruses in raw (untreated) and treated sewage water of a wastewater treatment plant in Greece”
P. Kokkinos · S. Filippidou ·K.Karlou· A.Vantarakis
Environmental Microbiology Unit, Department of Public Health, Medical School, University of Patras, Greece

The sewage treatment plant effluents may contain infectious human viruses representing a major public health issue. In the present study, a six months survey was conducted in order to evaluate the presence of enteroviruses (EV), adenoviruses (AdV), and hepatitis A viruses (HAV) in raw and treated sewage samples collected from a primary treatment wastewater plant, located at Alexandroupoli, in the North-Eastern Greece. Reverse transcriptase-polymerase chain reaction (RT-PCR) and nested polymerase chain reaction techniques have been applied for viral nucleic acid detection. Positive samples were confirmed by sequencing, and comparative phylogenetic analysis was performed on the isolated viral strains. EVs, AdVs, and HAV have been detected in 40% (10/25), 40% (10/25), 4% (1/25) of the samples collected from the inlet, and in 12% (3/25), 44% (11/25), 0% (0/25) of the samples collected from the outlet. Adenovirus types 3 (Ad3), 10 (Ad10) and 41 (Ad41), and hepatitis A virus type H2 have been recognized, while for enteroviruses Coxsackie type A2 and Echovirus types 27 and 30 have been recorded. The results reveal that treated sewage may still contain human viruses and thereby represent a potential health hazard. Moreover, this study shows the usefulness of molecular methods for virus detection, typing and virological quality analysis of sewage treatment plants.

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. Mavridou A, Smeti E, Mandilara G, Boufa P, Vagiona-Arvanitidou M, Vantarakis A, Vassilandonopoulou G, Pappa O, Roussia V, Tzouanopoulos A, Livadara M, Aisopou I, Maraka V, Nikolaou E. Technological Educational Institution […]

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.

Mavridou A, Smeti E, Mandilara G, Boufa P, Vagiona-Arvanitidou M, Vantarakis A, Vassilandonopoulou G, Pappa O, Roussia V, Tzouanopoulos A, Livadara M, Aisopou I, Maraka V, Nikolaou E.

Technological Educational Institution of Athens, Aigaleo 12210, Greece. amavridou@teiath.gr

“Recovery of melanized yeasts from Eastern Mediterranean beach sand associated with prevailing geochemical and marine flora patterns” Efstratiou,M.A. & Velegraki,A The melanized opportunistic pathogens Exophiala dermatitidis (Chaetothyriales) and Aureobasidium pullulans (Dothideales) were sporadically isolated from beach sand. This is the first time they are reported from bathing beach sand, thus providing updates on the conditions […]

“Recovery of melanized yeasts from Eastern Mediterranean beach sand associated with prevailing geochemical and marine flora patterns”
Efstratiou,M.A. & Velegraki,A

The melanized opportunistic pathogens Exophiala dermatitidis (Chaetothyriales) and Aureobasidium pullulans (Dothideales) were sporadically isolated from beach sand. This is the first time they are reported from bathing beach sand, thus providing updates on the conditions influencing the in situ black yeast community structure and raising public health concerns.

Do 2006/7/EC European Union Bathing Water Standards exclude the risk of contact with Salmonella or Candida albicans? Efstratiou,M.A. & Tsirtsis,G The purpose of this study was to evaluate the efficiency of the recent (2006) European Union Directive concerning the management of bathing water quality to exclude the presence of pathogens from complying waters. Coastal water […]

Do 2006/7/EC European Union Bathing Water Standards exclude the risk of contact with Salmonella or Candida albicans?
Efstratiou,M.A. & Tsirtsis,G

The purpose of this study was to evaluate the efficiency of the recent (2006) European Union Directive concerning the management of bathing water quality to exclude the presence of pathogens from complying waters. Coastal water samples were classified according to the Directive. 2.5% of ‘excellent’ water contained Salmonella and 39.2% Candida albicans 11.8% of samples in category ‘good’ were Salmonella positive and 35.2% were C.albicans positive. When the USEPA criterion for marine waters was applied to the same dataset, fewer samples complied, however 6.0% of the complying samples contained Salmonella. The results suggest that the bacterial indicator threshold levels for marine recreational water quality of the 2006/7/EC European Union Directive do not fully exclude contact of bathers with dangerous pathogens. Enterococci, if used as the sole index, appear to form a more reliable proxy of the risk of contact with Salmonella.

“Analysis of seawater microbiological quality data in Greece from 1997 to 2006: association of risk factors with bacterial indicators” P.Papastergiou, V.Mouchtouri, M.Karanika, E.Kostara, F.Kolokythopoulou, N.Mpitsolas, A.Papaioannou and C. Hadjichristodoulou This study deals with the examination of quality of seawater bathing areas in Greece over a 10-year period and identifies risk factors for high bacteria indicator organism […]

“Analysis of seawater microbiological quality data in Greece from 1997 to 2006: association of risk factors with bacterial indicators”
P.Papastergiou, V.Mouchtouri, M.Karanika, E.Kostara, F.Kolokythopoulou, N.Mpitsolas, A.Papaioannou and C. Hadjichristodoulou

This study deals with the examination of quality of seawater bathing areas in Greece over a 10-year period and identifies risk factors for high bacteria indicator organism concentrations. Qualitative descriptive analysis was applied and the microbiological test results of 231,205 water samples were associated with pollution markers and other parameters. Measurements of Escherichia coli (99.6%) and enterococci (100%) were found to be in  accordance with the mandatory value guidelines set by the new European Directive. An increasing trend for the yearly mean value of faecal streptococci was noted. Using logistic regression analysis, phenolic smell (OR Ό 2.10, CI Ό 2.04-2.16), rainfall the day before sampling (OR Ό 1.67, CI Ό 1.64-1.74), high seas (OR Ό 1.42, CI Ό 1.39-1.46) and rainfall on the day of sampling (OR Ό 1.27, CI Ό 1.20-1.33) were positively independently associated with high levels of bacterial indicators (total coliforms, faecal coliforms, faecal streptococci and E. coli). The highest risk, absolute risk value 42.8% (RR Ό 3.17, CI Ό 2.97-3.38), was measured when previous day rainfall, phenolic smell and high seas were simultaneously recorded. Such parameters should be further investigated as predetermining factors for the assessment of beach bathing water quality, providing a timely indication of water risk assessment.

“Prediction of Salmonella in seawater by total and faecal coliforms and Enterococci” Maria A. Efstratiou, Athena Mavridou, Clive Richardson The power of total coliforms (TC), faecal coliforms (FC) and Enterococci to predict the presence of Salmonella in seawater was investigated. Indicator cut-off values with the most satisfactory combination of sensitivity and specificity in predicting Salmonella […]

“Prediction of Salmonella in seawater by total and faecal coliforms and Enterococci”
Maria A. Efstratiou, Athena Mavridou, Clive Richardson

The power of total coliforms (TC), faecal coliforms (FC) and Enterococci to predict the presence of Salmonella in seawater was investigated. Indicator cut-off values with the most satisfactory combination of sensitivity and specificity in predicting Salmonella presence were 1000 CFU 100 ml¡1 TC, 200 CFU 100 ml¡1 FC, 500 CFU 100 ml¡1 Enterococci. When TC or FC were used for Salmonella prediction in logistic regression, then the addition of another indicator did not have a statistically significant effect. When Enterococci were  used for prediction, then the addition of either of the two other  indicators led to a statistically significant improvement (P = 0.001 for TC, P = 0.003 for FC). These results suggest that using either TC or FC alone provided an adequate indicator of Salmonella presence, but a statistically significant improvement is possible over using Enterococci alone. Concerning Enterococci, European Union limits for excellent coastal water quality (100 CFU 100 ml¡1) and United States  Environmental Protection Agency criteria for marine bathing waters (35 CFU 100 ml¡1) have the same value in predicting Salmonella absence (92.5%).

“Prevalence study of Legionella spp. contamination in Greek hospitals” A.Mavridou, E.Smeti, G.Mandilara, O.Pappa, S.Plakadonaki, E.Grispou and M.Polemis Water and swab samples were collected from 13 hospitals and analyzed for Legionella counts. Legionella was detected in eight out of 13 hospitals and in 22 of 130 water and swab-collected samples. A total of 72.7% of the […]

“Prevalence study of Legionella spp. contamination in Greek hospitals”
A.Mavridou, E.Smeti, G.Mandilara, O.Pappa, S.Plakadonaki, E.Grispou and M.Polemis

Water and swab samples were collected from 13 hospitals and analyzed for Legionella counts. Legionella was detected in eight out of 13 hospitals and in 22 of 130 water and swab-collected samples. A total of 72.7% of the strains were L. pneumophila ser. 1, 22.7% were L. pneumophila ser. 2-14, and 4.5% did not belong to any of these groups. AFLP typing of the L. pneumophila ser. 1 strains generated two distinguishable AFLP types. There was no significant correlation to the sample type with Legionella recovery. Legionella isolation was more likely to occur in the cooling towers than the water system. Water temperatures of 30-408C seem to favor Legionella growth. Of the 265 serum samples taken from the medical and technical staff for the control of IgG titre, 89.4% were negative, 7.2% were positive, and for 3.4% the result was doubtful. No association between IgG titre and maximum observed level of Legionella occurrence was detected.