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Background In a country-wide seroprevalence study of COVID-19 in Estonia, we aimed to determine the seroprevalence and the dynamics of IgG against SARS-CoV-2 after vaccination or positive PCR-test. Methods Leftover blood samples were selected between 8 February and 25 March 2021, by SYNLAB Estonia from all counties and age groups (0–9, 10–19, 20–59, 60–69, 70–79 and 80–100 years) proportionally to the whole Estonian population and tested for IgG against SARS-CoV-2 spike protein receptor-binding domain (anti-S-RBD IgG) using Abbott SARS-CoV-2 IgG II Quant assay. Antibody levels after positive PCR-test or vaccination were described by exponential increase-decrease models. Results According to total of 2517 samples, overall seroprevalence (95% confidence interval [CI]) was 20.1% (18.5–21.7%), similar in all age groups, but varied between counties. If individuals vaccinated with the first dose at least 14 d before antibody measurement were assumed to be seronegative, the overall seroprevalence was 15.8% (14.4–17.3%), 4.0-fold larger than the proportion of PCR-confirmed COVID-19 cases. Of seropositive individuals (n = 506) 194 (38.3%; 33.8–43.1%) had not had positive PCR-test or been vaccinated. According to exponential increase-decrease model, the peak of anti-S-RBD IgG in a 52-year-old (median age of PCR-positive and/or vaccinated individuals) was significantly higher after vaccination compared with positive PCR-test (22,082 (12,897–26,875) vs. 6732 (2321–8243) AU/mL), but half-life was similar (26.5 (6.9–46.1) vs. 38.3 (8.2–68.5) d). Conclusions One year after the start of COVID-19 pandemic the actual prevalence of infection is still underestimated compared with PCR-confirmed COVID-19 cases. Older compared with younger individuals have lower anti-S-RBD IgG level after vaccination, but similar decline rate.

The high number of mutations in the Omicron variant of SARS-CoV-2 cause its immune escape when compared to the earlier variants of concern (VOC). At least three vaccine doses are required for the induction of Omicron neutralizing antibodies and further reducing the risk for hospitalization. However, most of the studies have focused on the immediate response after the booster vaccination while the duration of immune response is less known. We here studied longitudinal serum samples from the vaccinated individuals up to three months after their third dose of the BNT162b2 vaccine for their capacity to produce protective antibodies and T cell responses to Wuhan and Omicron variants. After the second dose, the antibody levels to the unmutated spike protein were significantly decreased at three months, and only 4% of the individuals were able to inhibit Omicron spike interaction compared to 47%, 38%, and 14% of individuals inhibiting wild-type, delta, and beta variants’ spike protein. Nine months after the second vaccination, the antibody levels were similar to the levels before the first dose and none of the sera inhibited SARS-CoV-2 wild-type or any of the three VOCs. The booster dose remarkably increased antibody levels and their ability to inhibit all variants. Three months after the booster the antibody levels and the inhibition activity were trending lower but still up and not significantly different from their peak values at two weeks after the third dose. Although responsiveness towards mutated spike peptides was lost in less than 20 % of vaccinated individuals, the wild-type spike-specific CD4+ and CD8+ memory T cells were still present at three months after the booster vaccination in the majority of studied individuals. Our data show that two doses of the BNT62b2 vaccine are not sufficient to protect against the Omicron variant, however, the spike-specific antibodies and T cell responses are strongly elicited and well maintained three months after the third vaccination dose.

Background SARS-CoV-2 mRNA vaccines have proven high efficacy, however, limited data exists on the duration of immune responses and their relation to age and side effects. Methods We studied the antibody and memory T cell responses after the two-dose BNT162b2 vaccine in 122 volunteers up to 6 months and correlated the findings with age and side effects. Findings We found a robust antibody response to Spike protein after the second dose. However, the antibody levels declined at 12 weeks and 6 months post-vaccination, indicating a waning of the immune response over time. At 6 months after the second dose, the Spike antibody levels were similar to the levels in persons vaccinated with one dose or in COVID-19 convalescent individuals. The antibodies efficiently blocked ACE2 receptor binding to SARS-CoV-2 Spike protein of five variants of concern at one week but this was decreased at three months. 87% of individuals developed Spike-specific memory T cell responses, which were lower in individuals with increased proportions of immunosenescent CD8+ TEMRA cells. We found antibody response to correlate negatively with age and positively with the total score of vaccination side effects. Interpretation The mRNA vaccine induces a strong antibody response to SARS-CoV-2 and five VOCs at 1 week post-vaccination that decreases thereafter. T cell responses, although detectable in the majority, were lower in individuals with higher T cell immunosenescence. The deterioration of vaccine response suggests the need to monitor for the potential booster vaccination.

Purpose In Estonia, during the first wave of COVID-19 total number of cases confirmed by PCR was 13.3/10,000, similar in most regions, including capital Tallinn, but in the hotspot of Estonian epidemic, an island Saaremaa, the cumulative incidence was 166.1/10,000. We aimed to determine the prevalence of SARS-CoV-2 IgG antibodies in these two regions, symptoms associated with infection and factors associated with antibody concentrations. Methods Participants were selected using stratified (formed by age decades) random sampling and recruited by general practitioners. IgG or neutralizing antibodies were determined from sera by four assays. Symptoms associated with seropositivity were analyzed by multiple correspondence analysis, antibody concentrations by multiple linear regression. Results Total of 3608 individual were invited and 1960 recruited from May 8 to July 31, 2020. Seroprevalence was 1.5% (95% confidence interval (CI) 0.9–2.5) and 6.3% (95% CI 5.0–7.9), infection fatality rate 0.1% (95% CI 0.0–0.2) and 1.3% (95% CI 0.4–2.1) in Tallinn and Saaremaa, respectively. Of seropositive subjects 19.2% (14/73) had acute respiratory illness. Fever, diarrhea and the absence of cough and runny nose were associated with seropositivity in individuals aged 50 or more years. IgG, but not neutralizing antibodies concentrations were higher if fever, difficulty breathing, shortness of breath, chest pain or diarrhea was present, or hospitalization required. Conclusion Similarly to other European countries the seroprevalence of SARS-CoV-2 in Estonia was low even in the hotspot region Saaremaa suggesting that majority of population is susceptible to SARS-CoV-2. Focusing only on respiratory symptoms may delay accurate diagnosis of SARS-CoV-2 infection.

PurposeIn a country-wide seroprevalence study of COVID-19 in Estonia we aimed to determine the seroprevalence and the dynamics of IgG against SARS-CoV-2 after vaccination or positive PCR-test.MethodsLeftover blood samples were selected between February 8 to March 25, 2021, by SYNLAB Estonia from all counties and age groups (0-9, 10-19, 20-59, 60-69, 70-79, 80-100 years) proportionally to the whole Estonian population and tested for IgG against SARS-CoV-2 spike protein receptor-binding domain (anti-S-RBD IgG) using Abbott SARS-CoV-2 IgG II Quant assay. Antibody levels after positive PCR-test or vaccination were described by nonlinear model.ResultsA total of 2517 samples were tested. Overall seroprevalence (95% CI) was 20.1% (18.5-21.7%), similar in all age groups. If all individuals vaccinated with the first dose at least 14 days before antibody measurement were assumed to be seronegative, the overall seroprevalence was 15.8% (14.4-17.3%), 4-fold larger than the proportion of confirmed COVID-19 cases. According to nonlinear models, age increased anti-S-RBD IgG production after positive PCR-test but decreased after vaccination. The peak of anti-S-RBD IgG in a 52-year-old (median age of PCR-positive and/or vaccinated individuals) was significantly higher after vaccination compared with positive PCR-test (22082 (12897…26875) vs 6732 (2321…8243) AU/mL), but half-life was similar (26.5 (6.9…46.1) vs 38.3 (8.2…68.5) days).ConclusionOne year after the start of COVID-19 pandemic the actual prevalence of infection is still underestimated compared with confirmed COVID-19 cases, underlining the importance of seroepidemiological studies. Older individuals have lower anti-S-RBD IgG level after vaccination, but similar decline rate to younger.

The clinical features of SARS-CoV-2 infection range from asymptomatic to severe disease with life-threatening complications. Understanding the persistence of immune responses in asymptomatic individuals merit special attention because of their importance in controlling the spread of the infections. We here studied the antibody and T cell responses, and a wide range of inflammation markers, in 56 SARS-CoV-2 antibody-positive individuals, identified by a population screen after the first wave of SARS-CoV-2 infection. These, mostly asymptomatic individuals, were reanalyzed 7-8 months after their infection together with 115 age-matched seronegative controls. We found that 7-8 months after the infection their antibodies to SARS-CoV-2 Nucleocapsid (N) protein declined whereas we found no decrease in the antibodies to Spike receptor-binding domain (S-RBD) when compared to the findings at seropositivity identification. In contrast to antibodies to N protein, the antibodies to S-RBD correlated with the viral neutralization capacity and with CD4+ T cell responses as measured by antigen-specific upregulation of CD137 and CD69 markers. Unexpectedly we found the asymptomatic antibody-positive individuals to have increased serum levels of S100A12, TGF-alpha, IL18, and OSM, the markers of activated macrophages-monocytes, suggesting long-term persistent inflammatory effect associated with the viral infection in asymptomatic individuals. Our results support the evidence for the long-term persistence of the inflammation process and the need for post-infection clinical monitoring of SARS-CoV-2 infected asymptomatic individuals.

BackgroundAlthough the SARS-CoV-2 mRNA vaccines have proven high efficacy, limited data exists on the duration of immune responses and their relation to age and side effects.MethodsWe studied the antibody and memory T cell responses to Spike protein after the two-dose Comirnaty mRNA vaccine in 122 volunteers up to 3 months and correlated the findings with age and side effects.FindingsWe found a robust antibody response after the second vaccination dose. However, the antibody levels declined at 6 and 12 weeks postvaccination, indicating a waning of the immune response. Regardless, the average levels remained higher compared to pre-vaccination or in COVID-19 convalescent individuals. The antibodies efficiently blocked ACE2 receptor binding to Spike protein of four variants of concern at one week but this was decreased at three months, in particular with B.1.351 and P1 isolates. 87% of individuals developed Spike-specific memory T cell responses, which were lower in individuals with increased proportions of immunosenescent CD8+ TEMRA cells. We found a decreased vaccination efficacy but fewer adverse events in older individuals, suggesting a detrimental impact of age on outcome.InterpretationThe mRNA vaccine induces a strong antibody response to four variants at 1 week postvaccination but decreases thereafter, in particular among older individuals. T cell responses, although detectable in the majority, were lower in individuals with immunosenescence. The deterioration of vaccine response needs to be monitored to define the optimal time for the revaccination.FundingThe Estonian Research Council, Icosagen Cell Factory, and SYNLAB Estonia.Research in contextEvidence before this studyThe first studies addressing the immune responses in older individuals after the administration of SARS-CoV-2 mRNA vaccines have been published. We searched PubMed and medRxiv for publications on the immune response of SARS-CoV-2-mRNA vaccines, published in English, using the search terms “SARS-CoV-2”, “COVID-19”, “vaccine response”, “mRNA vaccine”, up to May 20th, 2021. To date, most mRNA vaccine response studies have not been peer-reviewed, and data on the dynamics of antibody response, role of age and side effects on SARS-CoV-2-mRNA vaccines in real vaccination situations is limited. Some studies have found a weaker immune response in older individuals after the first dose and these have been measured at a relatively short period (within one to two weeks) after the first dose but little longer-term evidence exists on the postvaccination antibody persistence.Added value of this studyIn this study, we assessed the antibody response up to three months after the full vaccination with Pfizer-BioNTech Comirnaty mRNA vaccine in 122 individuals. Our findings show strong Spike RBD antibody responses one week after the second dose with the capacity to block ACE2-Spike protein interaction, however, the antibodies declined significantly at three months after the second dose. The inhibition of ACE2-Spike interaction was weaker with South African (B.1.351) and Brazilian (P.1) than with Wuhan and UK (B.1.1.7) SARS-CoV-2 isolates. At three months 87% of vaccinated individuals developed either CD4+ or CD8+ T cell responses. Those negative for Spike-specific T cell response also tended to have lower Spike-specific antibody levels. In addition, CD4+ T cell response was decreased among vaccinated individuals with elevated levels of senescent CD8+ TEMRA cells. We found a weaker antibody response and faster waning of antibodies in older vaccinated individuals, which correlated with fewer side effects at the time of vaccinations.Implications of all the available evidenceOur results show that two doses of Pfizer-BioNTech Comirnaty mRNA vaccine induce a strong antibody and T cell responses to Spike RBD region but the antibody levels are declined at three months after the second dose. Nevertheless, even at three months, the anti-Spike RBD antibody levels stay significantly higher than at prevaccination, after the first dose of vaccine, or in Covid-19 postinfection. Our findings implicate older individuals to have fewer vaccination adverse effects and weaker immune response after the vaccination and point to the need for more individualized vaccination protocols, in particular among older people.

The emerging spread of variants of concern (VOC) of SARS-CoV-2 has been noted in several countries worldwide during last months. VOCs associated with increased transmissibility and morality. Sequencing is the gold standard for investigation of variants, however it is expensive and time-consuming. S-dropout routine monitoring in combination with VOC screening by RT-PCR is a useful tool for VOC surveillance.

The high number of mutations in the Omicron variant of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes its immune escape. We report a longitudinal analysis of 111 vaccinated individuals for their antibody levels up to 6 months after the third dose of the BNT162b2 vaccine. After the third dose, the antibody levels decline but less than after the second dose. The booster dose remarkably increases the serum ability to block wild-type or Omicron variant spike protein's receptor-binding domain (RBD) interaction with the angiotensin-converting enzyme 2 (ACE2) receptor, and these protective antibodies persist 3 months later. Three months after the booster dose, memory CD4

Variability in SARS-CoV-2 susceptibility and COVID-19 disease severity between individuals is partly due to genetic factors. Here, we identify 4 genomic loci with suggestive associations for SARS-CoV-2 susceptibility and 19 for COVID-19 disease severity. Four of these 23 loci likely have an ethnicity-specific component. Genome-wide association study (GWAS) signals in 11 loci colocalize with expression quantitative trait loci (eQTLs) associated with the expression of 20 genes in 62 tissues/cell types (range: 1:43 tissues/gene), including lung, brain, heart, muscle, and skin as well as the digestive system and immune system. We perform genetic fine mapping to compute 99% credible SNP sets, which identify 10 GWAS loci that have eight or fewer SNPs in the credible set, including three loci with one single likely causal SNP. Our study suggests that the diverse symptoms and disease severity of COVID-19 observed between individuals is associated with variants across the genome, affecting gene expression levels in a wide variety of tissue types., Graphical abstract, D’Antonio et al. characterize associations between GWAS signals for COVID-19 disease and eQTLs in 69 human tissues to identify causal variants and their underlying molecular mechanisms. They show that diverse symptoms and disease severity of COVID-19 are associated with variants affecting gene expression in a wide variety of tissues.

Background: Although the SARS-CoV2 mRNA vaccines have proven high efficacy in clinical trials, there is limited data on the impact of age, sex, and side effects on vaccine-induced immune responses. Methods: We studied the development of SARS-CoV-2 S-RBD antibodies after the Comirnaty mRNA vaccine in 118 healthy volunteers and correlated immune response with age, sex, and side effects of the vaccinations. Findings: We show a robust immune response to the S-RBD region after the first and the second vaccination dose. We saw a decline of antibody levels at 6 weeks versus 1 week after the second dose, suggesting a waning of the immune response over time. Regardless of this, the antibody levels at 6 weeks after the second dose remained significantly higher than before the vaccination, after the first dose, or in COVID-19 convalescent individuals. We found a decreased vaccination efficacy but fewer adverse events in older individuals, and that vaccination is less efficient in older males whereas the detrimental impact of age on vaccination outcome is abolished in females at 6 weeks after the second dose. Interpretation: The vaccine induces a strong immune response but older individuals develop fewer side effects and decreased antibody levels at 6 weeks. The waning of antibodies in particular in older male individuals suggests that both age and male sex act as risk factors in the immune response to the vaccine. Funding: The study was supported by the Centre of Excellence in Translational Genomics (EXCEGEN), the Estonian Research Council grant PRG377 and SYNLAB Estonia. Declaration of Interests: None to declare. Ethics Approval Statement: The study has been approved by the Research Ethics Committee of the University of Tartu on February 15, 2021 (nr 335/T-21).

BackgroundIn Estonia, during the first wave of COVID-19 total number of cases confirmed by PCR was 13.3/10,000, similar in most regions, including capital Tallinn, but in the hotspot of Estonian epidemic, an island Saaremaa, the cumulative incidence was 166.1/10,000.AimWe aimed to determine the prevalence of SARS-CoV-2 IgG antibodies in these two regions, symptoms associated with infection and factors associated with antibody concentrations.MethodsParticipants were selected using stratified (formed by age decades) random sampling and recruited by general practitioners. IgG were determined from sera by four assays. Symptoms of acute respiratory illness associated with seropositivity were analyzed by multiple correspondence analysis, antibody concentrations by multiple linear regression.ResultsTotal of 3608 individual were invited and 1960 recruited From May 8 to July 31, 2020. Seroprevalence was 1.5% (95% confidence interval (CI) 0.9-2.5) and 6.3% (95% CI 5.0-7.9), infection fatality rate 0.1% (95% CI 0.0-0.2) and 1.3% (95% CI 0.4-2.1) in Tallinn and Saaremaa, respectively. Of seropositive subjects 19.2% (14/73) had acute respiratory illness. Fever, diarrhea and the absence of cough and runny nose were associated with seropositivity in individuals aged 50 or more years. IgG concentrations were higher if fever, difficulty breathing, shortness of breath, chest pain or diarrhea was present, or hospitalization required.ConclusionSimilarly to other European countries the seroprevalence of SARS-CoV-2 in Estonia was low even in the hotspot region Saaremaa suggesting that majority of population is still susceptible to SARS-CoV-2. Focusing only on respiratory symptoms may delay accurate diagnosis of SARS-CoV-2 infection.

SARS-CoV-2 antibody tests are available in various formats, detecting different viral target proteins and antibody subclasses. The specificity and sensitivity of SARS-CoV-2 antibody tests are known to vary and very few studies have addressed the performance of these tests in COVID-19 patient groups at different time points. We here compared the sensitivity and specificity of seven commercial (SNIBE, Epitope, Euroimmun, Roche, Abbott, DiaSorin, Biosensor) and two in-house LIPS assays (LIPS N and LIPS S-RBD) IgG/total Ab tests in serum samples from 97 COVID-19 patients and 100 controls, and correlated the results with the patients' clinical data and the time-point the test was performed. We found a remarkable variation in the sensitivity of antibody tests with the following performance: LIPS N (91.8%), Epitope (85.6%), Abbott and in-house LIPS S-RBD (both 84.5%), Roche (83.5%), Euroimmun (82.5%), DiaSorin (81.4%), SNIBE (70.1%), and Biosensor (64.9%). The overall agreement between the tests was between 71-95%, whereas the specificity of all tests was within 98-100%. The correlation with patients' clinical symptoms score ranged from strongest in LIPS N (ρ = 0.41; p

Introduction The pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) has exposed vulnerable populations to an unprecedented global health crisis. The knowledge gained from previous human coronavirus outbreaks suggests that pregnant women and their fetuses are particularly susceptible to poor outcomes. The objective of this study was to summarize the clinical manifestations and maternal and perinatal outcomes of COVID‐19 during pregnancy. Material and methods We searched databases for all case reports and series from 12 February to 4 April 2020. Multiple terms and combinations were used including COVID‐19, pregnancy, maternal mortality, maternal morbidity, complications, clinical manifestations, neonatal morbidity, intrauterine fetal death, neonatal mortality and SARS‐CoV‐2. Eligibility criteria included peer‐reviewed publications written in English or Chinese and quantitative real‐time polymerase chain reaction (PCR) or dual fluorescence PCR‐confirmed SARS‐CoV‐2 infection. Unpublished reports, unspecified date and location of the study or suspicion of duplicate reporting, cases with suspected COVID‐19 that were not confirmed by a laboratory test, and unreported maternal or perinatal outcomes were excluded. Data on clinical manifestations, maternal and perinatal outcomes including vertical transmission were extracted and analyzed. Results Eighteen articles reporting data from 108 pregnancies between 8 December 2019 and 1 April 2020 were included in the current study. Most reports described women presenting in the third trimester with fever (68%) and coughing (34%). Lymphocytopenia (59%) with elevated C‐reactive protein (70%) was observed and 91% of the women were delivered by cesarean section. Three maternal intensive care unit admissions were noted but no maternal deaths. One neonatal death and one intrauterine death were also reported. Conclusions Although the majority of mothers were discharged without any major complications, severe maternal morbidity as a result of COVID‐19 and perinatal deaths were reported. Vertical transmission of the COVID‐19 could not be ruled out. Careful monitoring of pregnancies with COVID‐19 and measures to prevent neonatal infection are warranted.

SummaryBackgroundHigh number of SARS-CoV-2 antibody tests are available in different formats, detect different types of antibodies, and use different target proteins. Sensitivity of these tests varies and could be also related to clinical symptoms and testing time.MethodsSerum samples from 97 COVID-19 patients and 100 controls were tested with 9 antibody tests (SNIBE, Epitope, Euroimmun, Roche, Abbott, DiaSorin, Biosensor, LIPS N, and LIPS S-RBD). The results were analyzed in context of clinical data.FindingsPositivity rate was of tests was following: N-LIPS test (91.8% cases), Epitope (85.6%), Abbott and in-house LIPS S-RBD (both 84.5%), Roche (83.5%), Euroimmun (82.5%), DiaSorin (81.4%), SNIBE (70.1%), and Biosensor (64.9%). Agreement between tests varied (71-95%). Correlation between patient symptoms score and antibody value was test-dependent: varied from strongest in LIPS N (ρ=0.41; pInterpretationSensitivity of tests varied highly and combination of different tests may improve it. Relation of results to symptoms and testing time was test-dependent. Thus, some antibody tests seems to be more sensitive to detect antibodies early and in asymptomatic patients than others.FundingStudy was funded by SYNLAB Estonia and Estonian Research Council grant PRG377.Research in contextEvidence before this studyHigh variation in COVID-19 antibody tests sensitivity has been described. Relation between antibody response and clinical cause has been found in some studies but not in others. It’s known that antibody response is time dependent, however seroconversion medians varied in different studies.Added value of this studyWe confirmed that SARS-CoV-2 antibody response depends on clinical symptoms and time of testing, but we also found that this relation is dependent on test setup and viral antigens used in the tests. This may explain contradictory results of previous studies.Implications of all the available evidenceOur study has practical implications showing that not all antibody tests work uniformly well in symptomatic and asymptomatic cases and in different time periods from disease onset. This should be taken into consideration in clinical practice for diagnosing COVID-19 and in epidemiological studies evaluating the seroprevalence especially in asymptomatic population.

Clostridium difficile infection (CDI) is one of the most prevalent healthcare associated infections in hospitals and nursing homes. Different approaches are used for prevention of CDI. Absence of intestinal lactobacilli and bifidobacteria has been associated with C. difficile colonization in hospitalized patients. Our aim was to test a) the susceptibility of C. difficile strains of different origin and the intestinal probiotic Lactobacillus plantarum Inducia (DSM 21379) to various antimicrobial preparations incl. metronidazole, vancomycin; b) the susceptibility of C. difficile strains to antagonistic effects of the probiotic L. plantarum Inducia, prebiotic xylitol (Xyl) and their combination as a synbiotic (Syn) product; c) the suppression of germination of C. difficile spores in vitro and in vivo in animal model of C. difficile infection with Inducia, Xyl and Syn treatment. The VPI strain 10463 (ATCC 43255), epidemic strain (M 13042) and clinical isolates (n = 12) of C. difficile from Norway and Estonia were susceptible and contrarily L. plantarum Inducia resistant to vancomycin, metronidazole and ciprofloxacin. The intact cells of Inducia, natural and neutralized cell free supernatant inhibited in vitro the growth of tested C. difficile reference strain VPI and Estonian and Norwegian clinical isolates of C. difficile after co-cultivation. This effect against C. difficile sustained in liquid media under ampicillin (0.75 μg/ml) and Xyl (5%) application. Further, incubation of Inducia in the media with 5% Xyl fully stopped germination of spores of C. difficile VPI strain after 48 h. In infection model the 48 hamsters were administered ampicillin (30 mg/kg) and 10–30 spores of C. difficile VPI strain. They also received five days before and after the challenge a pretreatment with a synbiotic (single daily dose of L. plantarum Inducia 1 ml of 10 10 CFU/ml and 20% xylitol in 1 ml by orogastric gavage). The survival rate of hamsters was increased to 78% compared to 13% (p = 0.003) survival rate of hamsters who received no treatment. When administered Xyl the survival rate of hamsters reached 56% vs.13% (p = 0.06). In both Syn (6/9, p = 0.003) and Xyl (3/9, p = 0.042) groups the number of animals not colonized with C. difficile significantly increased. In conclusion, the combination of xylitol with L. plantarum Inducia suppresses the germination of spores and outgrowth into vegetative toxin producing cells of C. difficile and reduces the colonization of gut with the pathogen. Putative therapeutical approach includes usage of the synbiotic during antimicrobial therapy for prevention of CDI and its potential to reduce recurrences of CDI.

Selective reporting of antibiotic susceptibility test (AST) results is one possible laboratory-based antibiotic stewardship intervention. The primary aim of this study was to identify where and how selective reporting of AST results is implemented in Europe both in inpatient and in outpatient settings. An ESCMID cross-sectional, self-administered, internet-based survey was conducted among all EUCIC (European Committee on Infection Control) or EUCAST (European Committee on Antimicrobial Susceptibility Testing) national representatives in Europe and Israel. Of 38 countries, 36 chose to participate in the survey. Selective reporting of AST results was implemented in 11/36 countries (31%), was partially implemented in 4/36 (11%) and was limited to local initiatives or was not adopted in 21/36 (58%). It was endorsed as standard of care by health authorities in only three countries. The organisation of selective reporting was everywhere discretionally managed by each laboratory, with a pronounced intra-and inter-country variability. The most frequent application was in uncomplicated community-acquired infections, particularly urinary tract and skin and soft-tissue infections. The list of reported antibiotics ranged from a few first-line options, to longer reports where only last-resort antibiotics were hidden. Several barriers to implementation were reported, mainly lack of guidelines, poor system support, insufficient resources, and lack of professionals' capability. In conclusion, selective reporting of AST results is poorly implemented in Europe and is applied with a huge heterogeneity of practices. Development of an international framework, based on existing initiatives and identified barriers, could favour its dissemination as one important element of antibiotic stewardship programmes. KEYWORDS: Antibiotic prescription; Antibiotic resistance; Antibiotic stewardship; Antibiotic susceptibility testing; Questionnaire; Selective reporting PMID: 28093208 DOI: 10.1016/j.ijantimicag.2016.11.014, Selective reporting of antibiotic susceptibility test (AST) results is one possible laboratory-based antibiotic stewardship intervention. The primary aim of this study was to identify where and how selective reporting of AST results is implemented in Europe both in inpatient and in outpatient settings. An ESCMID cross-sectional, self-administered, internet-based survey was conducted among all EUCIC (European Committee on Infection Control) or EUCAST (European Committee on Antimicrobial Susceptibility Testing) national representatives in Europe and Israel. Of 38 countries, 36 chose to participate in the survey. Selective reporting of AST results was implemented in 11/36 countries (31%), was partially implemented in 4/36 (11%) and was limited to local initiatives or was not adopted in 21/36 (58%). It was endorsed as standard of care by health authorities in only three countries. The organisation of selective reporting was everywhere discretionally managed by each laboratory, with a pronounced intra- and inter-country variability. The most frequent application was in uncomplicated community-acquired infections, particularly urinary tract and skin and soft-tissue infections. The list of reported antibiotics ranged from a few first-line options, to longer reports where only last-resort antibiotics were hidden. Several barriers to implementation were reported, mainly lack of guidelines, poor system support, insufficient resources, and lack of professionals' capability. In conclusion, selective reporting of AST results is poorly implemented in Europe and is applied with a huge heterogeneity of practices. Development of an international framework, based on existing initiatives and identified barriers, could favour its dissemination as one important element of antibiotic stewardship programmes.

Avoiding food-borne diseases by competitive exclusion agents is a proactive strategy. In the current paper, we report the use of Bacillus smithii TBMI12 spores as potential competitive exclusion agents. One group of mice was predosed for three successive days with 108 colony forming units of B. smithii TBMI12 spores followed by inoculation with 106 colony forming units of wild-type Salmonella enterica serotype Enteritidis cells. Microbial plate counts of the animals' livers and spleens showed that only 40% of the mice were infected with S. enterica serotype Enteritidis, while the control group was 100% infected. These results suggest that B. smithii TBMI12 spores may protect against infection by S. enterica serotype Enteritidis.

Introduction. It is well established that severe periodontitis clusters in families, but there are no data about the relationship between mothers with chronic periodontitis and their children’s periodontal status. Objective. To evaluate a risk for periodontal diseases in children of periodontally diseased and healthy mothers. Methods. Four study groups were included: (I) 20 female patients with untreated generalized severe chronic periodontitis, (II) their children (34), (III) 13 periodontally healthy mothers and (IV) their children (13). Material was collected from years 2004–2006. The clinical examination included registration of visible plaque index, modified gingival index and, bleeding sites on probing. Periodontal microbiological samples were obtained from all study subjects and the isolates were identified according to morphology and biochemical profiles; similar interfamilial pathogens were compared by PCR-technique. Results. The children of diseased mothers more frequently had periodontal diseases, especially gingivitis. In addition, clinical parameters of gingival inflammation were more expressed and oral hygiene was worse in this group of children. VPI and VPI% of the diseased and healthy mothers differed significantly. The most common oral pathogens were P. intermedia/nigrescens and A. actinomycetemcomitans. The children of healthy mothers harboured pathogens less frequently than the children of diseased mothers. The sharing of P. intermedia/nigrescens was more frequent (5 families) than A. actinomycetemcomitans (2 families). Conclusion. Maternal indicators, such as periodontitis, hygiene habits, and periodontal microflora are risk factors for childhood periodontal diseases, and might be predictive of future childhood and adolescent periodontitis.

Background Fast, accurate and high-throughput detection of bacteria is in great demand. The present work was conducted to investigate the possibility of identifying both known and unknown bacterial strains from unassembled next-generation sequencing reads using custom-made guide trees. Results A program named StrainSeeker was developed that constructs a list of specific k-mers for each node of any given Newick-format tree and enables rapid identification of bacterial genomes within minutes. StrainSeeker has been tested and shown to successfully identify Escherichia coli strains from mixed samples in less than 5 minutes. StrainSeeker can also identify bacterial strains from highly diverse metagenomics samples. StrainSeeker is available at http://bioinfo.ut.ee/strainseeker. Conclusions Our novel approach can be useful for both clinical diagnostics and research laboratories because novel bacterial strains are constantly emerging and their fast and accurate detection is very important.

A total of 560 invasive and 1062 non-invasive isolates were collected. The antimicrobial susceptibility of invasive versus non-invasive Pseudomonas aeruginosa, Acinetobacter baumannii, and Klebsiella pneumoniae isolates were evaluated using the E-tests. The equal domination of Gram-negative among both invasive and non-invasive pathogens was estimated in our study if contaminants were excluded. The emergence trend of Gram-positive microbes especially of coagulase negative staphylococci may be proved only after application of exclusive algorithms. Due to similar susceptibility, the data of non-invasive Gram-negative pathogens can be useful to predict resistance of invasive ones. Also, the surveillance of invasive pathogens provides useful information about the general susceptibility of pathogens.

This prospective cohort study was performed from April to December 2003 for the purpose of collecting a maximum of 50 non-duplicate isolates of Acinetobacter baumannii, Pseudomonas aeruginosa, and Klebsiella pneumoniae from each of 4 ICUs to determine minimum inhibitory concentrations. The most prevalent species were Enterobacteriaceae (13%), K. pneumoniae and A. baumannii (both 12%). 60% of A. baumannii strains were susceptible to ampicillin/sulbactam and cefepime, 95% to meropenem and imipenem, and 75% to amikacin. 79% of P. aeruginosa strains were piperacillin/tazobactam, 58% ceftazidime, 81% meropenem, 72% imipenem, 69% ciprofloxacin and 97% amikacin susceptible. The susceptibility of K. pneumoniae to meropenem and imipenem was 99%, to ciprofloxacin was 91% and to amikacin was 98%. Gram-negative bacteria (especially K. pneumoniae and A. baumannii) were prevalent in our ICUs compared to other European studies. Carbapenem susceptibility of Estonian strains was higher, but P. aeruginosa sensitivity to ceftazidime was lower, compared to other EU countries.

The objectives of this study were to comprehensively characterize the range, content, and performance of sexually transmitted infection (STI) testing services in Estonia during the period 2001 to 2002 and to determine if the observed diagnostic laboratory practices and methods adhered to international evidence-based recommendations.Survey data, focusing on organization and performance characteristics of STI diagnostics services, were assessed using questionnaires, telephone interviews, and site visits to all responding facilities providing STI diagnostics services in Estonia. Guidelines of international evidence-based recommendations for STI testing were used as references.There were significant shortcomings in STI testing availability and practices. Among all participating laboratories diagnosing STIs, only a minority (n = 16, 28%) offered testing for the full minimum range of relevant STIs in Estonia, i.e., Treponema pallidum, Neisseria gonorrhoeae, Chlamydia trachomatis, and Trichomonas vaginalis. In addition, because testing methods used were not properly selected, confirmation of several infections in accordance with evidence-based requirements was not possible, which has an impact both on STI diagnostic quality and surveillance.

Indigenous intestinal microflora (including lactobacilli) has an important role in protection against Clostridium difficile infection. To assess in vitro interaction between lactobacilli and C. difficile, antagonistic activity of 50 intestinal Lactobacillus spp. strains against 23 pathogenic C. difficile strains was determined. Phenotypic properties of C. difficile strains [production of short-chain fatty acids (SCFAs) and toxin A, and antimicrobial susceptibility] and lactobacilli (production of SCFAs and H(2)O(2)) were investigated. Five lactobacilli (Lactobacillus paracasei and Lactobacillus plantarum species) were antagonistic to all C. difficile strains, 18 were antagonistic to some C. difficile strains and 27 showed no antagonistic activity. This antagonistic activity was strain-specific and seemed to correlate with H(2)O(2) and lactic acid production. C. difficile strains that were more sensitive to lactobacilli (n = 9) usually produced higher toxin levels and more SCFAs, and were more resistant to antibiotics, than strains that were resistant to lactobacilli (n = 14). Compatibility of C. difficile strain properties (resistance to lactobacilli or antibiotics) with intestinal microecological conditions (e.g. presence of antagonistic lactobacilli, concentration of antibiotics) may determine expression of disease.

At present little or no data is available regarding the resistance profiles of anaerobic bacteria in relation to the general usage of antibiotics. The objective of this study was to assess whether any potential relationship exists between the dynamics of antibiotic resistance of anaerobic bacteria and the consumption of antibiotics during the last 3 years within the Estonian population. In total, 416 anaerobic isolates were investigated from various clinical samples. The anaerobes were isolated on Wilkins-Chalgren Agar, incubated in an anaerobic glove box and identified by standard methods. β -lactamase negative strains were tested against metronidazole, clindamycin, benzylpenicillin and the positive strains were further tested against metronidazole, clindamycin, and ampicillin/sulbactam by E-tests. The results of the susceptibility tests were interpreted according to the current criteria of NCCLS. Data from the Estonian State Agency of Medicines was used to assess the antibiotic consumption rate in the population (Defined Daily Doses per 1000 inhabitants annually). The following species of anaerobes were isolated: B. fragilis group, Bacteroides sp., Fusobacterium sp., Porphyromonas sp., Prevotella sp., Peptostreptococcus sp., in addition to various unidentified Gram-positive rods. Metronidazole resistance was not found among Gram-negative bacteria despite a relatively high consumption of this antimicrobial agent in Estonia. Only ampicillin/sulbactam demonstrated excellent in vitro activity against all anaerobes. Unexpectedly despite a relatively low rate of consumption of clindamycin a high rate of resistance to this agent occurred; a similar situation was noted for penicillin. In the present study we did not observe a relationship between the changes in antibiotic consumption (DDD/1000) rate and the resistance pattern of anaerobic bacteria to metronidazole, clindamycin, penicillin and ampicillin/sulbactam during a 3-year follow-up period. High resistance to penicillin among some species and also to clindamycin is similar to the global trend and argues for limited use of these antibiotics in empirical treatment. We would suggest that monitoring of local susceptibility pattern is necessary for the selection of initial empirical therapy.

The antibiotic and chlorhexidine (CHX) susceptibility of 70 distinct clinical isolates: Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumannii, Staphylococcus aureus (not MRSA), Streptococcus pyogenes and Enterococcus faecalis (10 of each) were tested using minimal bactericidal (MBC) and/or minimal inhibitory (MIC) concentrations. Non-fermentative bacteria tolerated CHX at high concentrations; Gram-positive cocci, especially S. pyogenes, were the most susceptible. We found a good correlation between CHX and antibiotic susceptibility in both MIC and MBC among Gram-negative bacteria, and mainly in MBC among Gram-positive bacteria. Resistance to ciprofloxacin, imipenem, cefotaxime, ceftazidime, gentamicin and aztreonam appeared to indicate increased CHX resistance among Gram-negative bacteria. This finding gives clinicians the ability to predict CHX susceptibility according to routine antibiotic resistance testing.

Comparing culture- and non-culture-based methods for quantifying Clostridium difficile in antibiotic-associated-diarrhea patients, we found that the real-time PCR method correlated well with quantitative culture and was more sensitive. A positive association between the population levels of C. difficile and the presence of its toxins was found.

The aim of this study was to compare the prevalence of different virulence factor (VF) genes in extended-spectrum beta-lactamase (ESBL) producingEscherichia colistrains isolated from the Baltic Sea region. A total of 432 strains of phenotypically ESBL positiveE. coliwere collected from 20 institutions located in Estonia, Latvia, Lithuania, and the region of St. Petersburg in Russia from January to May 2012 and analyzed for phylogenetic group and prevalence of 23 VF genes. The strains were collected from clinical material (urine, blood, wound, and respiratory tract). Bacterial isolates were compared according to phylogenetic group, clinical material, and geographical origin. Most of the VF genes were concentrated within phylogenetic group B2 and/or D. When comparing strains isolated from different countries, it was found that strains originating from Estonia and Latvia belonged mainly to group B2 and strains from Lithuania and Russia mainly to groups B2 and D. The P-fimbrial adhesin genepapEFwas more prevalent in Russian strains, colicin genecvaCin Lithuanian strains, and capsular genekpsMTIIin Latvian strains; serum resistant genetraTwas less prevalent in Estonian strains. The regional differences of VF genes remained statistically significant after taking into account the phylogenetic distribution in the countries.

The use of antibiotics, type of infections and resistance of prevalent bacteria was surveyed in Tartu University hospitals. The data from two ICUs (1995 and 1998), surgical and internal medicine departments (1998) were compared. Overall antibiotic usage in the ICUs and in the hospital as a whole had increased. There was a significant increase in Gram-positive bacterial infections and a decrease in Gram-negative infections in the ICUs. At the same time, susceptibility to several antibiotics decreased in most of the prevalent Gram-negative aerobes in the ICUs (Acinetobacter spp., Pseudomonas spp., Klebsiella spp.). Exceptions to this were the greater susceptibility of Pseudomonas spp. to gentamicin and Acinetobacter spp. to imipenem. Some changes in the predominant bacterial populations did not correlate to changes in antibiotic use.

Little is known about possible regional differences in the development of the intestinal microflora in infants. The aim of the present study was therefore to compare the development of the microflora in Estonian and Swedish infants during the first month of life. The intestinal microflora of one week old and one month old Estonian ( n= 20) and Swedish ( n= 20) infants was studied by quantitative culture of faecal samples. All newborns were delivered vaginally and fed with breast milk during the first month of life. At one week of age the counts of aerobes (coagulase negative staphylococci, enterococci, enterobacteria) were higher in Estonian than in Swedish newborns. The counts of lactobacilli and candida increased in the Estonian infants over the first month of life, while the counts of enterobacteria increased in Swedish infants. At one month of age, the Estonian infants were more frequently colonised with lactobacilli than the Swedish infants and the counts were higher. Our data indicate that previously described differences in intestinal microflora of Estonian and Swedish 1 to 2 year old children are present already at the first month of life. Keywords: newborn, intestinal microflora, geographic differences.

Background: Laboratory diagnosis of syphilis is usually accomplished by serology. There are currently a large number of different commercial treponemal tests available that vary in format, sensitivity and specificity. Aim: To evaluate the ID-PaGIA Syphilis Antibody Test as an alternative to other specific treponemal tests for primary screening or confirmation of diagnosis. Methods: Serum samples from healthy adults (n = 100) were used for detection of specificity of ID-PaGIA. To evaluate sensitivity of ID-PaGIA serum samples (n = 101) from patients with confirmed or suspected syphilis were tested for syphilis antibodies with FTA-Abs IgM, ID-PaGIA, ELISA IgM and TPHA tests. Results: No false-positive results were found with ID-PaGIA. Sensitivity of various treponemal tests was the following: FTA-Abs IgM: 95.5%, ID-PaGIA and ELISA IgM: 94%, and TPHA 75%. The positive and negative predictive values of ID-PaGIA were 100 and 89.5%, respectively. Conclusions: Compared with other treponemal tests ID-PaGIA has excellent sensitivity and specificity.

Background The prevalence of allergic diseases seems to have increased particularly over the past 35–40 years. Furthermore, allergic disease is less common among children in the formerly socialist countries of central and Eastern Europe as compared with Western Europe. It has been suggested that a reduced microbial stimulation during infancy and early childhood would result in a slower postnatal maturation of the immune system and development of an optimal balance between TH1- and TH2-like immunity. Aims To test the hypothesis that allergic disease among children may be associated with differences in their intestinal microflora in two countries with a low (Estonia) and a high (Sweden) prevalence of allergy. Methods From a prospective study of the development of allergy in relation to environmental factors, 29 Estonian and 33 Swedish 2-year-old children were selected. They were either nonallergic (n = 36) or had a confirmed diagnosis of allergy (n = 27) as verified by typical history and at least one positive skin prick test to egg or cow's milk. Weighed samples of faeces were serially diluted (10−2–10−9) and grown under anaerobic conditions. The counts of the various genera and species were calculated for each child. In addition, the relative amounts of the particular microbes were expressed as a proportion of the total count. Results The allergic children in Estonia and Sweden were less often colonized with lactobacilli (P

The spread of carbapenemase-producing Enterobacteriaceae is a global problem; however, no exact data on the epidemiology of carbapenemase in the Baltic countries and St. Petersburg area is available. We aimed to evaluate the epidemiology of carbapenemase-producingEscherichia coliandKlebsiella pneumoniaein the Baltic States and St. Petersburg, Russia, and to compare the different methods for carbapenemase detection. From January to May 2012, allK. pneumoniae n=1983andE. coli n=7774clinical isolates from 20 institutions in Estonia, Latvia, Lithuania, and St. Petersburg, Russia were screened for carbapenem susceptibility. The IMP, VIM, GIM, NDM, KPC, and OXA-48 genes were detected using real-time PCR and the ability to hydrolyze ertapenem was determined using MALDI-TOF MS. Seventy-seven strains were found to be carbapenem nonsusceptible. From these, 15K. pneumoniaestrains hydrolyzed ertapenem and carried theblaNDMgene. All of these strains carried integron 1 and most carried integron 3 as well as genes of the CTX-M-1 group. No carbapenemase-producingE. coliorK. pneumoniaestrains were found in Estonia, Latvia, or Lithuania; however, NDM-positiveK. pneumoniaewas present in the hospital in St. Petersburg, Russia. A MALDI-TOF MS-based assay is a suitable and cost-effective method for the initial confirmation of carbapenemase production.

Few studies provide rates of nosocomial bloodstream infections (BSIs) in mixed neonatal and paediatric intensive care units (PICUs).To determine the rate, pathogens and outcome of BSIs in an Estonian PICU.Data were collected prospectively from 1st January 2004 to 31st December 2008 in the PICU of Tartu University Hospital. The definition criteria of the US Centers for Disease Control and Prevention were applied for the diagnosis of laboratory-confirmed BSI.A total of 126 episodes of BSI were identified in 89 patients (74 neonates, eight infants, seven patients aged1 year). Among neonates 42 (57%) had birth weight1000 g. The overall incidence of BSI was 9.2 per 100 admissions, incidence density 12.8 per 1000 patient-days. Primary BSI was diagnosed in 92 episodes. Central line (CL)-associated BSI incidence density for neonates was 8.6 per 1000 CL-days with the highest incidence (27.4) among neonates with extremely low birth weight. The most common pathogens were coagulase-negative staphylococci (43%) and Serratia marcescens (14%). Resistance to meticillin was detected in four out of seven S. aureus isolates (all were part of an outbreak) and 23% of Enterobacteriaceae were extended spectrum beta-lactamase (ESBL)-producing strains. Overall case-fatality rate was 10%.We observed higher rates of BSIs in our mixed PICU than reported previously. High levels of antimicrobial resistance were detected. Future research should focus on the effects of infection control measures to prevent outbreaks and to decrease incidence of CL-associated BSI.

The disruption of intestinal microbiota is an important risk factor for the development of Clostridium difficile caused antibiotic associated diarrhea (AAD). The role of intestinal lactoflora in protection against C. difficile is unclear. Fecal samples (n = 74) from AAD patients were investigated for C. difficile and lactobacilli by culture and real-time PCR. Lactobacilli were identified by enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) and sequencing of 16S rRNA. In C. difficile negative cases we found somewhat higher counts of intestinal Lactobacilli (5.02 vs. 2.15 CFU log(10)/g; p = 0.053) by culture and more frequently Lactobacillus plantarum (33.3% vs. 9.4%; p = 0.03) as compared with positive ones. Results of total counts of lactobacilli comparing Estonian and Norwegian samples were conflicting by culture and PCR. We found higher colonization of Norwegian AAD patients with L. plantarum (21% vs. 5%, p = 0.053) and Estonians with Lactobacillus gasseri (19% vs. 2%, p = 0.023). Particular lactobacilli (e.g. L. plantarum) may have a role in protection against C. difficile, whereas the meaning of total counts of lactobacilli remains questionable. In different persons and nations, different lactobacilli species may have a protective role against C. difficile.

Purpose: To report initial results from a European ICU surveillance programme focussing on antibiotic consumption, microbial resistance and infection control. Methods: Thirty-five ICUs participated during 2005. Microbial resistance, antibiotic consumption and infection control stewardship measures were entered locally into a web-application. Results were validated locally, aggregated by project leaders and fed back to support local audit and benchmarking. Results: Median (range) antibiotic consumption was 1,254 (range 348–4,992) DDD per 1,000 occupied bed days. The proportion of MRSA was median 11.6% (range 0–100), for ESBL phenotype of E. coli and K. pneumoniae 3.9% (0–80) and 14.3% (0–77.8) respectively, and for carbapenem-resistant P. aeruginosa 22.5% (0–100). Screening on admission for alert pathogens was commonly omitted, and there was a lack of single rooms for isolation. Conclusions: The surveillance programme demonstrated wide variation in antibiotic consumption, microbial resistance and infection control measures. The programme may, by providing rapid access to aggregated results, promote local and regional audit and benchmarking of antibiotic use and infection control practices. The original publication is available at www.springerlink.com: Håkan Hanberger, Dilek Arman, Hans Gill, Vlastimil Jindrák, Smilja Kalenic, Andrea Kurcz, Monica Licker, Paul Naaber, Elizabeth A. Scicluna, Václav Vanis and Sten M. Walther, Surveillance of microbial resistance in European Intensive Care Units: a first report from the Care-ICU programme for improved infection control, 2009, (35), 1, 91-100, Intensive Care Medicine. Copyright: Springer-Verlag, www.springerlink.com

A prospective multicentre hospital-wide surveillance study was performed to investigate nosocomial bloodstream infections (BSIs) and to promote BSI surveillance in Estonia in 2004-2005. All patients from the acute care departments of two referral centres and one central hospital were included. A total of 549 episodes of BSI occurred in 507 patients (0.6 cases per 1000 patient-days). Of those, 55% occurred in intensive care units and 47% were catheter-associated infections. Of BSI cases, 24% occurred in patients with haematological malignancy. The in-hospital case-fatality rate was 31%. Of causative micro-organisms, 315 (53%) were Gram-positive aerobes, 232 (39%) were Gram-negative aerobes and 35 (6%) were fungi. Anaerobic bacteria accounted for 2%. The most common pathogens were coagulase-negative staphylococci (26%), Enterobacteriaceae (24%), enterococci (13%) and pseudomonas (10%). Eight percent of BSI were polymicrobial. Seven percent of Staphylococcus aureus isolates were meticillin resistant. Of pseudomonas isolates, 19%, 25%, 30% and 44% were resistant to ceftazidime, meropenem, piperacillin/tazobactam and imipenem, respectively. The incidence of BSI did not differ significantly from other reported studies. With the exception of relatively high antimicrobial resistance among pseudomonas, the overall resistance patterns of Estonian nosocomial bloodstream pathogens were similar to those seen in Nordic countries and lower than in Central and Southern Europe. This study contributes to the development and implementation of surveillance in Estonian hospitals.

Our study confronts the use of antimicrobial agents in ambulatory care with the resistance trends of 2 major pathogens, Streptococcus pneumoniae and Escherichia coli, in 21 European countries in 2000–2005 and explores whether the notion that antimicrobial drug use determines resistance can be supported by surveillance data at national aggregation levels. The data obtained from the European Surveillance of Antimicrobial Consumption and the European Antimicrobial Resistance Surveillance System suggest that variation of consumption coincides with the occurrence of resistance at the country level. Linear regression analysis showed that the association between antimicrobial drug use and resistance was specific and robust for 2 of 3 compound pathogen combinations, stable over time, but not sensitive enough to explain all of the observed variations. Ecologic studies based on routine surveillance data indicate a relation between use and resistance and support interventions designed to reduce antimicrobial drug consumption at a national level in Europe., peer-reviewed

In 28 adult severe periodontitis patients who did not respond to conventional periodontal therapy, full mouth clinical parameters including probing pocket depth, relative attachment level, bleeding on probing and suppuration after probing, visible plaque index and modified gingival index were recorded at the baseline and 14 months after treatment. Based on clinical and bacteriological diagnosis, a combination of systemic amoxicillin 500 mg x 3 and metronidazole 200 mg x 2 was prescribed for 7 days. In combination with non-surgical treatment, systemic antibiotic therapy, significantly improved median values of probing pocket depth, bleeding on probing, suppuration index, visible plaque index and modified gingival index except relative attachment level. Despite the improvement of clinical parameters in general, both bleeding on probing and suppuration index had significantly lower reduction in smokers than in non-smokers.

Our objective was to investigate the presence of periodontal pathogens in comparison with the total degree of microorganisms after non-surgical periodontal therapy. The study material consisted of microbiological samples from periodontal pockets originating from 140 consecutive patients with chronic generalized severe periodontitis. The subgingival samples from periodontal pockets were obtained by a sterile curette, placed into 2 ml of the VMGA III medium, homogenized and serially diluted in the Brucella broth. 100 microl aliquots from the dilutions were inoculated onto the Brucella and the TSBV agar. The plates were incubated in an anaerobic chamber and under microaerobic conditions. The isolates were identified according to colonial and cellular morphology, the potency disk pattern, and the biochemical profiles. After instrumentation, no periodontal pathogens were isolated in 46 (33%) patients, while 94 patients (67%) were infected with one to five different periodontal pathogens. However, higher degree of the total microflora was positively correlated with number of isolated pathogens, a putative indicator of their presence. Therefore, due to the occurrence of residual microorganisms after non-surgical mechanical treatment, information about the pattern of these pathogens is needed for application of antimicrobial therapy. The level of microbial load in gingival pockets, including both pathogenic and non-pathogenic species, is one of the determinants of presence of residual pathogens after non-surgical periodontal therapy.