Jenny Kremastinou

Distribution of Serogroups and Genotypes among Disease-Associated and Carried Isolates of Neisseria meningitidis from the Czech Republic, Greece, and Norway

ABSTRACT The distribution of serogroups and multilocus sequence types (STs) in collections of disease-associated and carried meningococci from the period 1991 to 2000 in three European countries (the Czech Republic, Greece, and Norway) was investigated. A total of 314 patient isolates and 353 isolates from asymptomatic carriers were characterized. The frequency distributions of serogroups and clone complexes differed among countries and between disease and carrier isolate collections. Highly significant differentiation was seen at each housekeeping locus. A marked positive association of serogroup C with disease was evidenced. The ST-11 complex was strongly positively associated with disease; associations for other clone complexes were weaker. The genetic diversity of the clone complexes differed. A single ST dominated the ST-11 clone complex, while the ST-41/44 complex exhibited greater levels of diversity. These data robustly demonstrated differences in the distribution of meningococcal genotypes in disease and carrier isolates and among countries. Further, they indicated that differences in genotype diversity and pathogenicity exist between meningococcal clone complexes.

Economic Recession and Emergence of an HIV-1 Outbreak among Drug Injectors in Athens Metropolitan Area: A Longitudinal Study

Background During 2011, a dramatic increase (1600%) of reported HIV-1 infections among injecting drug users (IDUs) was noted in Athens, Greece. We herein assess the potential causal pathways associated with this outbreak. Methods Our study employed high resolution HIV-1 phylogenetic and phylogeographic analyses. We examined also longitudinal data of ecological variables such as the annual growth of gross domestic product (GDP) of Greece in association with HIV-1 and HCV sentinel prevalence in IDUs, unemployment and homelessness rates and HIV transmission networks in Athens IDUs before and during economic recession (2008–2012). Results IDU isolates sampled in 2011 and 2012 suggested transmission networks in 94.6% and 92.7% of the cases in striking contrast with the sporadic networking (5%) during 1998–2009. The geographic origin of most HIV-1 isolates was consistent with the recently documented migratory waves in Greece. The decline in GDP was inversely correlated with annual prevalence rates of HIV and HCV and with unemployment and homelessness rates in IDUs (all p<0.001). The slope of anti-HCV prevalence in the sentinel populations of IDUs and in “new” drug injectors was found 120 and 1.9-fold (p = 0.007, p = 0.08 respectively) higher in 2008–2012 (economic recession) compared with 2002–2006. The median (25th, 75th) size of transmission networks were 34 (12, 58) and 2 (2, 2) (p = 0.057) in 2008–2012 and 1998–2007, respectively. The coverage of harm reduction services was low throughout the study period. Conclusions Scaling-up harm reduction services and addressing social and structural factors related to the current economic crisis should be urgently considered in environments where HIV-1 outbreaks may occur.

Risk Factors for Contamination of Hotel Water Distribution Systems by Legionella Species

ABSTRACT The Legionella colonization frequency at 385 Greek hotel hot and cold water distribution systems was 20.8%. Legionella contamination was associated with the presence of an oil heater (odds ratio [OR] = 2.04, 95% confidence interval [CI] = 1.12 to 3.70), with the sample temperature (OR = 0.26, 95% CI = 0.1 to 0.5), with seasonal operation (OR = 3.23, 95% CI = 1.52 to 6.87), and with the presence of an independent disinfection system (OR = 0.30, 95% CI = 0.15 to 0.62). The same water temperatures, free-chlorine levels, and pHs differently affect the survival of various Legionella spp.

Factors affecting carriage of Neisseria meningitidis among Greek military recruits.

Greek military recruits (993) were examined for carriage of meningococci during July 1990. Blood, saliva and throat swab specimens were obtained and each recruit answered a questionnaire providing information on age, education (a measure of socioeconomic level), place of residence, smoking habits and recent infections. The overall carriage rate was 25% but differed between the two camps: 79/432 (18%) in Camp A and 168/561 (30%) in Camp B (P less than 0.0005). In Camp B, there were significantly higher proportions of recruits who were non-secretors (P less than 0.0005), and/or heavy smokers (P less than 0.0005). They were also younger (less than 19 years old) (P less than 0.001), and on the whole had fewer years of education (P less than 0.0005). By univariate analysis, carriage was significantly associated with smoking. By multiple logistic regression analysis, carriage was associated with smoking (P less than 0.001), age (P less than 0.01) and the camp in which the recruits were based (P less than 0.01). Among recruits in Camp B, 15/38 (40%) of those with recent viral infections were carriers compared with 30% for the camp in general.

Evaluation of non-culture diagnosis of invasive meningococcal disease by polymerase chain reaction (PCR)

Antibiotic treatment prior to transport or admission to hospital has reduced the proportion of cases of invasive meningococcal disease (IMD) from which Neisseria meningitidis can be isolated by standard microbiological techniques. Identification of meningococci by polymerase chain reaction (PCR) was assessed in relation to microbiological diagnosis for cases over a 4-year period between 1998 and 2001. A screening assay for the IS1106 gene was used to detect meningococcal DNA and five additional assays for siaD and orf-2 genes were performed to determine the serogroup. PCR results were compared with results of bacteriological culture, other laboratory test results and clinical data. The sensitivity of the PCR assay for culture-confirmed cases was 98.5%. The specificity of the assay was 96% based on test results for patients from whom other bacteria were isolated, children with viral meningitis and afebrile negative controls. The siaD B/C/W-135 and Y as well as the orf-2 gene for serogroup A PCR assays were able to determine the serogroup for 75.2% of cases that were positive by PCR screening assay. When isolates from patients with IMD were tested by both agglutination and PCR, the results agreed in all cases. PCR is a useful tool for diagnosis of IMD when Gram stain and culture tests are negative due to antibiotic treatment prior to collection of samples for microbiological analyses.

West Nile virus outbreak in humans, Greece, 2012: third consecutive year of local transmission

In 2010, the first outbreak of West Nile virus (WNV) infection in Greece was recorded, the largest in Europe since 1996. After 2010, outbreaks continued to occur in different areas of the country. Enhanced surveillance was implemented during transmission periods (June to October). We investigated the 2012 outbreak to determine its extent and identify risk factors for severe disease using regression models. Of 161 cases recorded in 2012, 109 had neuroinvasive disease (WNND). Two outbreak epicentres were identified: the southern suburbs of Athens in July and a rural area in East Macedonia T 95% CI: 2.2-22) and chronic renal failure (adjusted RR: 4.5; 95% CI: 2.7-7.5) were independently associated with WNND-related death. In three PCR-positive samples, sequencing revealed WNV lineage 2 identical to the 2010 strain. The occurrence of human cases in three consecutive years suggests that WNV lineage 2 has become established in Greece. Raising awareness among physicians and susceptible populations (elderly people and persons with co-morbidities) throughout Greece is critical to reduce the disease impact. .

Serogroups, serotypes and subtypes of Neisseria meningitidis isolated from patients and carriers in Greece

The increase in the number of cases of meningococcal disease reported to the Ministry of Health in Athens since 1989 prompted the present study to determine if isolates from patients or carriers expressed the same phenotypic characters as those in other parts of Europe. None of the isolates from patients (31) or carriers (547) expressed the antigenic combinations associated with outbreaks in northern Europe, i.e., B:15:P1.16 or B:4:P1.15. The majority of the Greek isolates did not react with any of the six monoclonal serotype reagents tested; however, most reacted with one or more of the 11 monoclonal subtype antibodies. The results suggest that additional serotype reagents are needed for epidemiological studies in southeastern Europe and that vaccines based on serotype antigens developed against outbreak strains in northern Europe would not be effective in Greece.

Molecular and Morphological Characterization of Aedes albopictus in Northwestern Greece and Differentiation from Aedes cretinus and Aedes aegypti

Abstract The presence of Aedes albopictus (Skuse) was recently confirmed for the first time in northwestern Greece. This location is within the distribution range of a morphologically similar species, Aedes cretinus Edwards, and is a potentially favorable region for the reintroduction of Aedes aegypti (L.). It was thus compelling to use methods in addition to morphology-based keys to correctly identify specimens badly damaged, rubbed, or otherwise altered in their external characteristics. It was decided to use molecular techniques as a novel and reliable method for differentiating the three Stegomyia species. The nuclear internal transcribed spacer 2 (ITS2) fragments from morphologically identified Ae. albopictus and Ae. cretinus specimens were amplified, and their sequences were compared with those in GenBank for Ae. albopictus, Ae. cretinus, and Ae. aegypti. Also, mitochondrial cytochrome oxidase I (COI) fragments were amplified for Ae. albopictus and Ae. cretinus (so far not available in GenBank) and compared with Ae. aegypti fragments. ITS2 and COI sequences generated in our study were deposited in GenBank and could be useful in future studies of mosquitoes by other research workers.

Antibiotic sensitivities of Neisseria meningitidis isolates from patients and carriers in Greece

Usage of antibiotics in southern Europe is less well regulated than in northern countries. The proportion (48%) of meningococci in Spain insensitive to pencillin (MIC greater than or equal to 0.1 mg/l) prompted this investigation of antibiotic sensitivities of isolates from Greek patients with meningitis (31) and carriers (47 school-children and 472 recruits). The agar dilution method was used to determine MIC to penicillin G (PN), sulphamethoxazole (SU), rifampicin (RF), cefaclor (CF) and ciprofloxacin (CP). The proportion of isolates insensitive to PN was 48% for isolates from patients, 19% from school-children and 36.6% from recruits. Resistance to SU (MIC greater than or equal to 16 mg/l) was found in 16% of those from patients, 10.6% from children and 40% from recruits. None of the isolates from patients was resistant to RF (greater than or equal to 1 mg/l) but 6% of those from carriers were. Resistance to CF (greater than or equal to 4 mg/l) was found in 9.2% of patient isolates, 6.4% from children and 23.7% from recruits. All isolates except one were sensitive to CP (MIC range less than 0.0015-0.125 mg/l). Resistances to PN, SU and RF were analysed by serogroup, serotype and subtype of the bacteria. The proportion of resistant isolates showed some variation between different areas of Greece, but it was not statistically significant.

Parental smoking and carriage of Neisseria meningitidis among Greek schoolchildren.

In December 1990 and January 1991, primary (320) and secondary (697) pupils in 2 areas of Athens were screened to determine the rate of carriage of Neisseria meningitidis and to determine if the genetic and environmental factors associated with carriage of meningococci in Greece were similar to those observed for northern European populations. In 1 area, socioeconomic indicators were significantly lower than in the other (p < 0.0005), but the isolation rates from pupils in the areas were similar, 5.3% and 6.3%. In contrast to studies in northwest Europe, carriage was not associated with lower socioeconomic conditions, sex, numbers of individuals per household, upper respiratory tract infection, or secretor status. By univariate analysis, carriage was associated with age (15-18 years) (p < 0.05) and mothers or other carers smoking habits (p < 0.05)--but not fathers smoking. Although the proportion of fathers who smoked was greater in the area where socioeconomic indicators were lower (61%) vs. (47%) (p < 0.0005), the proportions of women smokers were similar (33% vs. 38%). By multiple regression analysis, the only significant factors were age (p < 0.01) and carers smoking (p < 0.05).