Coagulase negative staphylococcal sepsis in neonates: do we need to adapt vancomycin dose or target?

BACKGROUND: Despite differences in types of infection and causative organisms, pharmacokinetic-pharmacodynamic (PKPD) targets of vancomycin therapy derived from adult studies are suggested for neonates. We aimed to identify doses needed for the attainment of AUC/MIC > 400 and AUC/MIC > 300 in neonates with sepsis and correlate these targets with recommended doses and treatment outcome. METHODS: Neonates who had Vancomycin therapeutic drug monitoring (TDM) performed between January 1, 2010 and December 31, 2012 were studied. Clinical characteristics, episodes of Gram-positive sepsis with outcomes and all neonatal blood culture isolates in hospital were collected from medical records. To estimate probability of target attainment of AUC/MIC >400 and AUC/MIC >300 a 1000-subject Monte Carlo simulation was performed by calculating AUC using Anderson's (Anderson et al. 2006) and TDM trough concentrations (Ctrough) based population PK models. RESULTS: Final dataset included 76 patients; 57 with confirmed Gram-positive sepsis. TDM was taken after the 1st to 44th dose. 84.1% of Ctrough were within the range 5-15 mg/L. Currently recommended doses achieved probability of the targets (PTA) of AUC/MIC >400 and AUC/MIC >300 in less than 25% and 40% of cases, respectively. Doses required for 80% PTA of AUC/MIC > 400 for MIC ≥2 mg/L resulted in Ctrough values ≥14 mg/L. Mean AUC/MIC values were similar in treatment failure and success groups. CONCLUSION: With currently recommended vancomycin dosing the therapeutic target of AUC/MIC > 400 is achieved only by 25% of neonates. Appropriate PKPD targets and respective dosing regimens need to be defined in prospective clinical studies in this population.

Higher blood glucose level associated with body mass index and gut microbiota in elderly people.

BACKGROUND: Some dominant bacterial divisions of the intestines have been linked to metabolic diseases such as overweight and diabetes. OBJECTIVE: A pilot study aimed to evaluate the relations between the culturable intestinal bacteria with body mass index (BMI) and some principal cellular and metabolic markers of blood in people older than 65. DESIGN: Altogether 38 generally healthy elderly people were recruited: ambulatory (n=19) and orthopedic surgery (n=19). Questionnaires on general health, anthropometric measurements, routine clinical and laboratory data, and quantitative composition of cultivable gut microbiota were performed. RESULTS: Blood glucose level was positively correlated with BMI (r=0.402; p=0.014). Higher blood glucose level had negative correlation with relative share of intestinal anaerobic bacteria such as bacteroides (r=-0.434; p=0.0076) and gram-positive anaerobic cocci (r=-0.364; p=0.027). In contrast, the relative share of bifidobacteria (r=0.383; p=0.019) and staphylococci (r=0.433; p=0.008) was positively correlated to blood glucose level. In elderly people, a higher blood glucose concentration was predicted by the reduction of the anaerobes' proportion (adj. sex, age, and BMI R(2)=0.192, p=0.028) and that of Bacteroides sp. (adj. R(2)=0.309, p=0.016). CONCLUSION: A tight interplay between increased BMI, level of blood glucose, and the reduced proportion of cultivable bacteroides is taking place in the gut microbiota of elderly people.

The association of gut microbiota with body weight and body mass index in preschool children of Estonia.

BACKGROUND: The gut microbiota has been shown to affect both fat storage and energy harvesting, suggesting that it plays a direct role in the development of obesity. The aim of this study was to investigate whether intestinal colonization by particular species/groups of the intestinal microbiota is related to body weight values in Estonian preschool children born in different years during the entire 1990s. METHODS: Body weight, height, body mass index (BMI), and quantitative composition of cultivable gut microbiota (staphylococci, enterococci, streptococci, enterobacteria, lactobacilli, anaerobic gram-positive cocci, bifidobacteria, eubacteria, bacteroides, clostridia, and candida) were studied in 51 healthy 5-year-old children (40 were born between 1993 and 94 and 11 were born between 1996 and 97). RESULTS: At the age of 5 years, median weight was 19.5 kg and median BMI was 15.3 kg/m(2). Significantly higher BMI (p=0.006) was found in 5-year-old children born in late versus early 1990s during the development of socioeconomic situation of Estonia (2% rise in gross domestic product). The counts of the different gut bacteria did not show any association with weight and BMI in the 5-year-old children. However, the BMI values were in positive correlation with a relative share of anaerobic gram-positive bacteria, for example, bifidobacteria when adjusted for sex and year of birth (adj R(2)=0.459, p=0.026) and eubacteria (adj R(2)=0.484, p=0.014) in the community of cultured intestinal microbiota. The relative share of bacteroides showed a negative correlation with the childrens' weight (adj R(2)=- 0.481, p=0.015). CONCLUSION: The body weight indices of preschool children of the general population are associated with the proportion of anaerobic intestinal microbiota and can be predicted by sex and particular socioeconomic situation from birth to 5 years of age.

In vitro synergy of oxacillin and gentamicin against coagulase-negative staphylococci from blood cultures of neonates with late-onset sepsis.

Coagulase-negative staphylococci (CoNS) are the leading cause of late-onset sepsis (LOS) in neonates. Increasing resistance of CoNS to beta-lactams and aminoglycosides has led to widespread use of vancomycin, which in turn may lead to resistance to vancomycin. Thus, combination therapy of LOS has been advocated. We aimed to determine the interaction of oxacillin and gentamicin against CoNS. In 2005, 34 isolates of oxacillin- and gentamicin-resistant CoNS were obtained from blood samples of neonates with LOS. Combination effect was tested using the checkerboard method, E-test with the other antibiotic incorporated in the medium (E-test-1) and two E-test strips placed in a cross-formation (E-test-2). Of 34 isolates 61.8%, 53% and 73.5% revealed synergy or an additive effect when tested by the checkerboard method, E-test-1 and E-test-2, respectively. Results of all three tests were concordant for six (17.6%) isolates, four showing synergy, and two indifference. Our in vitro results support that combination therapy with penicillinase-resistant penicillin and aminoglycoside can be an alternative to vancomycin.

Quantification of Clostridium difficile in antibiotic-associated-diarrhea patients.

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.

Intestinal lactoflora in Estonian and Norwegian patients with antibiotic associated diarrhea.

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.

Diversity and metabolic impact of intestinal Lactobacillus species in healthy adults and the elderly.

The present study aimed at assessing the counts and species distribution of intestinal lactobacilli and exploring if the data are associated with BMI and blood glucose level in healthy adults and elderly persons. The BMI (P < 0·01), the level of fasting blood glucose (P < 0·001) and the total counts of lactobacilli (P < 0·01 by bacteriology; P < 0·001 by real-time PCR) were higher in the elderly. The number of species in adults was lower (P < 0·05), who were more often colonised with Lactobacillus acidophilus (P = 0·031) and L. helveticus (P < 0·001). In contrast, L. plantarum (P = 0·035), L. paracasei (P < 0·001) and L. reuteri (P = 0·031) were more prevalent in the elderly. L. rhamnosus was detected in adults (P < 0·001), but not in any elderly person. BMI was associated with counts of lactobacilli, adjusted for age and sex (P = 0·008). The higher BMI in both groups of persons was associated with the presence of obligate homofermentative lactobacilli and L. sakei, both adjusted for age and sex. Plasma glucose values were positively correlated with BMI and negatively correlated with colonisation with L. paracasei (P = 0·0238) in adults and on the borderline with L. fermentum (P = 0·052) in the elderly. Thus, the species-specific PCR analysis of Lactobacillus sp. combined with viable plating data indicates substantial age-related structural differences in the intestinal lactobacilli communities. The higher counts of intestinal Lactobacillus sp. are associated with higher BMI and blood glucose content, while their specific fermentative groups and species of lactobacilli appear at different glucose levels both in adults and in the elderly.

Intestinal Lactobacillus sp. is associated with some cellular and metabolic characteristics of blood in elderly people.

The higher counts or particular groups (Firmicutes/Bacteroidetes) of intestinal microbiota are related to host metabolic reactions, supporting a balance of human ecosystem. We further explored whether intestinal lactobacilli were associated with some principal cellular and metabolic markers of blood in 38 healthy >65-year-old persons. The questionnaire, routine clinical and laboratory data of blood indices as much as the oxidized low-density lipoprotein (ox-LDL) and baseline diene conjugates in low-density lipoprotein (BDC-LDL) of blood sera were explored. The PCR-based intestinal Lactobacillus sp. composition and counts of cultivable lactobacilli (LAB) were tested. The facultative heterofermentative lactobacilli (Lactobacillus casei and Lactobacillus paracasei) were the most frequent (89 and 97%, respectively) species found, while Lactobacillus acidophilus, Lactobacillus plantarum and Lactobacillus reuteri were present in almost half of the elderly persons. The number of species simultaneously colonizing the individuals ranged from 1 to 7 (median 4). In elderly consuming probiotics the LAB counts were significantly higher than in these not consuming (median 7.8, range 4.2-10.8 vs. median 6.3, range 3.3-9.7 log cfu/g; p=0.005), adjusted (OR=1.71, CI95 1.04-2.82; p=0.035) for age and body mass index (BMI). The colonization by L. acidophilus was negatively related (r=-0.367, p=0.0275) to L. reuteri, staying significant after adjusting for age, sex and BMI (OR=0.16, CI95 0.04-0.73; p=0.018). However, the blood glucose concentration showed a tendency for a negative correlation for colonization with Lactobacillus fermentum (r=-0.309, p=0.062) adjusted for BMI (Adj. R(2)=0.181; p=0.013) but not for age and sex. The higher white blood cells (WBC) count was positively related (r=0.434, p=0.007) to presence of Lactobacillus reuteri adjusted for age, sex and BMI (Adj. R(2)=0.193, p=0.027). The lower values of ox-LDL were predicted by higher counts of cultivable lactobacilli adjusted by sex, age and BMI (r = -0.389, p = 0.016; Adj. R(2)=0.184 p=0.029). In conclusion, the pilot study of elderly persons shows that the intestinal lactobacilli are tightly associated with WBC count, blood glucose and content of ox-LDL which all serve as risk markers in pathogenesis of inflammation, metabolic syndrome and cardiovascular disease (CVD).

Periodontal disease in mothers indicates risk in their children.

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.

The occurrence of antimicrobial resistance and class 1 integrons among commensal Escherichia coli isolates from infants and elderly persons.

BACKGROUND: The aim of our study was to compare the presence of the intI1 gene and its associations with the antibiotic resistance of commensal Escherichia coli strains in children with/without previous antibiotic treatments and elderly hospitalized/healthy individuals. METHODS: One-hundred-and-fifteen intestinal E. coli strains were analyzed: 30 strains from 10 antibiotic-naive infants; 27 from 9 antibiotic-treated outpatient infants; 30 from 9 healthy elderly volunteers; and 28 from 9 hospitalized elderly patients. The MIC values of ampicillin, cefuroxime, cefotaxime, gentamicin, ciprofloxacin, and sulfamethoxazole were measured by E-test and IntI1 was detected by PCR. RESULTS: Out of the 115 strains, 56 (49%) carried class 1 integron genes. Comparing persons without medical interventions, we found in antibiotic-naive children a significantly higher frequency of integron-bearing strains and MIC values than in healthy elderly persons (53% versus 17%; p < 0.01). Evaluating medical interventions, we found a higher resistance and frequency of integrons in strains from hospitalized elderly persons compared with non-hospitalized ones. Children treated with antibiotics had strains with higher MIC values (when compared with antibiotic-naive ones), but the integron-bearing in strains was similar. In most cases, the differences in resistance between the groups (integron-positive and negative strains separately) were higher than the differences between integron-positive and negative strains within the groups. CONCLUSION: The prevalence of integrons in commensal E. coli strains in persons without previous medical intervention depended on age. The resistance of integron-carrying and non-carrying strains is more dependent on influencing factors (hospitalization and antibiotic administration) in particular groups than merely the presence or absence of integrons.

Evaluation of ID-PaGIA syphilis antibody test.

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.

Prevalence and antibiotic susceptibility of Acinetobacter baumannii, Pseudomonas aeruginosa and Klebsiella pneumoniae in Estonian intensive care units in comparison with European data.

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 comparison of susceptibility patterns of Gram-negative invasive and non-invasive pathogens in Estonian hospitals.

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.

The efficacy of non-surgical and systemic antibiotic treatment on smoking and non-smoking periodontitis patients.

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.

The microbiological status of patients with periodontitis in southern Estonia after non-surgical periodontal therapy.

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.

Helicobacter pylori substantially increases oxidative stress in indomethacin-exposed rat gastric mucosa.

Helicobacter pylori (H. pylori) often play an important role in the pathogenesis of gastritis, peptic ulcer, and probably also gastric cancer. Reactive oxygen species (ROS) produced by this bacterium may be one of the crucial factors whereby oxidative stress can play a role in the pathogenesis of ulcer disease. The aim of this study was to assess ROS activity and glutathione redox status, a principal cellular redox sensor, in H. pylori-associated indomethacin-induced gastric ulcers in rats. Gastric lesion was produced by intragastric administration of indomethacin (7 mg/kg) for three days followed by administration of H. pylori suspension (density 10(9) colony forming units). Animals receiving indomethacin only or followed by administration of H. pylori suspension were sacrificed after 11 and 18 days. ROS activity was assessed by the level of lipid peroxidation (LPO) and the glutathione redox status by the ratio between oxidized and reduced glutathione (GSSG/GSH). Indomethacin did not significantly increase the level of LPO and the GSSG/GSH ratio. When H. pylori suspension was given together with indomethacin the LPO was increased both on days 11 and 18 and GSSG/GSH on day 18. H. pylori, thus, substantially increases glutathione redox ratio and lipid peroxidation in gastric mucosa, which may play an important role in the pathological mechanisms of this bacterium. The findings support the idea that dietary antioxidants could be beneficial in combination therapy for eradication of H. pylori.

The diagnostic value of endoscopy and Helicobacter pylori tests for peptic ulcer patients in late post-treatment setting.

BACKGROUND: Guidelines for management of peptic ulcer patients after the treatment are largely directed to detection of H. pylori infection using only non-invasive tests. We compared the diagnostic value of non-invasive and endoscopy based H. pylori tests in a late post-treatment setting. METHODS: Altogether 34 patients with dyspeptic complaints were referred for gastroscopy 5 years after the treatment of peptic ulcer using a one-week triple therapy scheme. The endoscopic and histologic findings were evaluated according to the Sydney classification. Bacteriological, PCR and cytological investigations and 13C-UBT tests were performed. RESULTS: Seventeen patients were defined H. pylori positive by 13C-UBT test, PCR and histological examination. On endoscopy, peptic ulcer persisted in 4 H. pylori positive cases. Among the 6 cases with erosions of the gastric mucosa, only two patients were H. pylori positive. Mucosal atrophy and intestinal metaplasia were revealed both in the H. pylori positive and H. pylori negative cases. Bacteriological examination revealed three clarithromycin resistant H. pylori strains. Cytology failed to prove validity for diagnosing H. pylori in a post-treatment setting. CONCLUSIONS: In a late post-treatment setting, patients with dyspepsia should not be monitored only by non-invasive investigation methods; it is also justified to use the classical histological evaluation of H. pylori colonisation, PCR and bacteriology as they have shown good concordance with 13C-UBT. Moreover, endoscopy and histological investigation of a gastric biopsy have proved to be the methods with an additional diagnostic value, providing the physician with information about inflammatory, atrophic and metaplastic lesions of the stomach in dyspeptic H. pylori positive and negative patients. Bacteriological methods are suggested for detecting the putative antimicrobial resistance of H. pylori, aimed at successful eradication of infection in persistent peptic ulcer cases.

The limit of leucocytospermia from the microbiological viewpoint.

The aim of the study was to find out the correlation between white blood cell (WBC) counts in semen and quantitative composition of seminal microflora, and to establish the minimum WBC count associated with significant bacteriospermia. The research included 159 men with different WBC counts in their semen, 84 of them with chronic prostatitis/chronic pelvic pain syndrome. Semen samples were cultivated quantitatively for detecting anaerobic, microaerophilic and aerobic bacteria. Bryan-Leishman stained slides were used for detecting WBC in semen. Seminal fluid was colonized by eight different microorganisms, and the total count of microorganisms in semen ranged from 102 to 107 CFU ml-1. A high frequency of anaerobic microorganisms was found. A positive correlation was observed between the WBC count and the number of different microorganisms, and also between the WBC count and the total count of microorganisms in semen sample. The receiver operating characteristic curve analysis demonstrated that the WHO-defined WBC cut-off point (1 x 106 WBC ml-1) has very low sensitivity for discriminating between patients with and without significant bacteriospermia, as a more optimal sensitivity/specificity ratio appears at 0.2 x 106 WBC ml-1 of semen. The quantitative microbiological finding of semen in the patients of National Institute of Health (NIH) categories IIIa and IV was very similar, i.e. a high number of different microorganisms and a high total count of microorganisms. In the control group (without leucocytospermia and prostatitis symptoms) both parameters were significantly lower.

Antibiotic susceptibility of clinically relevant anaerobes in Estonia from 1999 to 2001.

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. beta-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.

Anaerobic seminal fluid micro-flora in chronic prostatitis/chronic pelvic pain syndrome patients.

We investigated the seminal micro-flora of 116 men. Eighty-four men had chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS), and 34 of them were also leukocytospermic. Thirty-two asymptomatic men formed the control group. Micro-organisms were found in all of the 116 seminal fluid specimens. More than 20 different micro-organisms were found in both groups. Neisseria gonorrhoeae and Chlamydia trachomatis were not found. A high frequency of anaerobic bacteria was found in all groups (68-79%), and in most of the specimens, anaerobic micro-organisms were equal to or outnumbered the aerobic strains. We found 1-8 different micro-organisms in each semen sample, the total count of micro-organisms ranged from 10(2) to 10(7)/mL of semen. Both parameters were significantly higher in leukocytospermic CP/CPPS (NIH IIIA category) patients (median=5 different micro-organisms; total median count 5 x 10(4)) than in the control group (median=3 different micro-organisms; total median count 10(3)). In the CP/CPPS patients, the prevalence and/or count of some opportunistic bacteria was higher than in the control group. To show that the micro-organisms do not originate from the urethra, first voided urine was also investigated in 17 prostatitis patients and 15 controls. One patient had significantly fewer micro-organisms (median 1 vs. 4) and a lower total count of micro-organisms (median 10(2) vs. 10(4)/mL) in the first-catch urine than in the seminal fluid. We found only one third of the micro-organisms to be similar in urine and semen while anaerobic bacteria and some aerobic opportunists were infrequent in urine. Semen is a suitable specimen for the diagnosis of prostatitis.