Impact of Lactiplantibacillus plantarum Inducia on metabolic and antioxidative response in cholesterol and BMI variable indices: randomised, double-blind, placebo-controlled trials.

Probiotics may have potential in reducing cardiovascular disease (CVD) risk in middle-aged persons with borderline metabolic indices. The ability of potential probiotic Lactiplantibacillus plantarum Inducia to reduce CVD risk factors in persons with variable cholesterol and body mass indices (BMI) was assessed. In two parallel-armed double-blind placebo-controlled interventions (n=136) and (n=104), participants daily received either test yoghurt (Inducia) or placebo yoghurt. BMI, blood pressure, plasma glucose, cholesterol, high-sensitivity C-reactive protein (hs-CRP), oxidative stress and immunological markers were measured. Total counts of lactobacilli and L. plantarum Inducia were evaluated using real-time PCR. Significant reduction of total cholesterol, low density lipoprotein cholesterol (LDL-c) and non-high-density cholesterol occurred in both trials. The change in cholesterol (P=0.023) in persons with normal BMI and borderline cholesterol levels after four weeks of yoghurt consumption was detected. A difference was also found between placebo and test yoghurt groups (P=0.042) in LDL-c with normal BMI. Blood glucose reduction (P=0.01) and antioxidative effect was detected in overweight volunteers of the test yoghurt group. The suppression of oxidised LDL was associated with lowered oxidative stress index and total peroxide concentration values and faecal recovery of Inducia. The Inducia strain expresses antioxidative effect on blood lipids and has anti-glycaemic impact that allow to apply it as dietary probiotic supplement for the management of CVD risks in humans.

A combination of the probiotic and prebiotic product can prevent the germination of Clostridium difficile spores and infection.

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 1010 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.

Impact of probiotic Lactobacillus plantarum TENSIA in different dairy products on anthropometric and blood biochemical indices of healthy adults.

The blood pressure-lowering effect of dairy products holds the potential to decrease the risk of cardiovascular disease (CVD). An open question is if the successful expression of functional properties of the probiotic strain depends on host biomarkers and/or food matrix properties. The probiotic Lactobacillus plantarum strain TENSIA® (DSM 21380) is a novel microorganism with antimicrobial and antihypertensive functional properties. The aim of this study was to characterise the functional properties of the probiotic L. plantarum TENSIA and compare its effects on host anthropometric, clinical, and blood biomarkers when consumed with cheese or yoghurt. This study involved two double-blinded randomised placebo-controlled exploratory trials (ISRCTN15061552 and ISRCTN79645828) of healthy adults over a three-week period. The three-week consumption of probiotic L. plantarum TENSIA in a daily dose of 1×1010 cfu in probiotic cheese or a daily dose of 6×109 cfu in yoghurt with different content of carbohydrates, proteins, and lipids did not significantly change the body mass index (BMI), plasma glucose and lipid levels, or inflammatory markers in the blood. Reduced lowered systolic and diastolic blood pressure values were detected, regardless of food matrix or baseline values for blood pressure and BMI. In conclusion, our study showed that three-week consumption of the probiotic L. plantarum TENSIA either in cheese or yoghurt lowered diastolic and systolic blood pressure regardless of food matrix and baseline values of blood pressure and BMI, confirming the impact of the functional properties of the probiotic strain in decreasing CVD risk.

Safety of a probiotic cheese containing Lactobacillus plantarum Tensia according to a variety of health indices in different age groups.

Safety of the probiotic Lactobacillus plantarum strain Tensia (DSM 21380) was tested in vitro, in semihard Edam-type cheese, in an animal model and after consumption of the probiotic cheese in double-blind randomized placebo-controlled human intervention studies with different age groups. The susceptibility of L. plantarum Tensia to 8 antibiotics, and the presence of tetracycline (tet M, S, O, K, L) genes and class 1 integron was assessed by applying epsilometer-test and PCR-based methods. Production of biogenic amines by the probiotic strain in decarboxylation medium containing 1% of l-histidine, l-glutamine, l-ornithine, l-arginine, or l-lysine and in cheese was tested by gas chromatography. The biosafety of L. plantarum Tensia was evaluated on National Institutes of Health-line mice fed cheese containing Tensia at a concentration of 9.6 log cfu/g for 30 consecutive days. In human intervention trials in adults and the elderly, the effects of different doses of Edam-type cheese and the probiotic bacterium on BW, gut functionality indices, and host metabolism were evaluated. The strain L. plantarum Tensia was susceptible to all tested antibiotics and did not possess the tetracycline resistance-determining genes tet(L), tet(S) and tet(O), nor did it contain the integron (Int1) gene. However, the strain was tet(K) and tet(M) positive. Lactobacillus plantarum Tensia did not produce potentially harmful biogenic amines, such as histamine or cadaverine. The amount of tyramine produced in the cheese environment during ripening and after 15 wk of storage was below the clinically significant content. In the animal model, no translocation of the administered strain or other microbes into the blood or organs of mice was detected. No harmful effect was observed on body mass index, inflammatory markers, or serum lipidograms during human intervention trials with different age groups at a daily dose of 10.3 or 8.17 log cfu/serving for 3 wk. No negative effect on gastrointestinal welfare was observed, but the consumption of 100g/d for 3 wk caused hard stools from the second week of the trial. The content of total lactobacilli increased in feces, and the presence of the ingested probiotic strain was confirmed after the consumption of cheese. Thus, L. plantarum strain Tensia is suitable for generally recognized as safe (GRAS) and qualified presumption of safety (QPS) criteria because it did not have any undesirable characteristics. The regular semihard Edam-type cheese (fat content of 26%) with the probiotic additive at a daily dose of 50 g or in excess (100g) and with a probiotic daily dose of 10 log cfu for 3 wk was safe.

[Effect of probiotic Heart cheese "Harmony" comprising bacteria L. plantarum TENSIA DSM 21380 on blood pressure and metabolic health indices].

The probiotic L. plantarum strain TENSIA (DSM 21380) is a novel microorganism having antimicrobial and antihypertensive properties. The aim of the study was to test the efficacy of the consumption of the cheese, comprising the novel strain TENSIA on multiple health markers of humans. Human intervention trial showed that the blood pressure lowering effect of cheese, comprising L. plantarum TENSIA was evident in healthy volunteers with high normal blood pressure up to normal values. The 3-week consumption of the prohiotic cheese did not increase the CVD risk factors like BMI, the level of plasma lipids and glucose as well as inflammatory and immunological markers of human body.

Safety and persistence of orally administered human Lactobacillus sp. strains in healthy adults.

The aim of the study was to evaluate the safety and persistence of selected Lactobacillus strains in the gastrointestinal tract (GIT) of healthy adult volunteers after oral consumption of high doses of lactobacilli to identify potential candidates for probiotic and biotechnological applications. In the first phase of the study, nine individuals consumed capsules containing Lactobacillus gasseri 177 and E16B7, Lactobacillus acidophilus 821-3, Lactobacillus paracasei 317 and Lactobacillus fermentum 338-1-1 (each daily dose 1×1010 cfu) for 5 consecutive days. Data on gut health, blood parameters, and liver and kidney function were collected. The persistence of Lactobacillus strains was assessed by culturing combined with arbitrarily primed polymerase chain reaction (AP-PCR) and PCR-denaturing gradient gel electrophoresis (PCR-DGGE) on days 0, 5, 8, 10 and 20 from faecal samples. All strains survived gastrointestinal passage and were detected on the 5th day. L. acidophilus 821-3 was detected in four volunteers on the 8th day (4.3 to 7.0 log10 cfu/g) and in two on the 10th day (8.3 and 3.9 log10 cfu/g, respectively). In the second phase of the study, five additional volunteers consumed L. acidophilus 821-3 (daily 1×1010 cfu) for 5 consecutive days. The strain was subsequently detected in faeces of all individuals using real-time PCR on the 10th day (range 4.6-6.7; median 6.0 log10 cell/g) in both phases of the study for at least 5 days after discontinuation of consumption. The administration of high doses of different Lactobacillus strains did not result in any severe adverse effects in GIT and/or abnormal values of blood indices. Thus, the strain L. acidophilus 821-3 is a promising candidate for probiotic and biotechnological applications. Further studies will be performed to confirm the strain persistence and safety in a larger number of individuals.

Complete glutathione system in probiotic Lactobacillus fermentum ME-3.

There is much information about glutathione (GSH) in eukaryotic cells, but relatively little is known about GSH in prokaryotes. Without GSH and glutathione redox cycle lactic acid bacteria (LAB) cannot protect themselves against reactive oxygen species. Previously we have shown the presence of GSH in Lactobacillus fermentum ME-3 (DSM14241). Results of this study show that probiotic L. fermentum ME-3 contains both glutathione peroxidase and glutathione reductase. We also present that L. fermentum ME-3 can transport GSH from environment and synthesize GSH. This means that it is characterized by a complete glutathione system: synthesis, uptake and redox turnover ability that makes L. fermentum ME-3 a perfect protector against oxidative stress. To our best knowledge studies on existence of the complete glutathione system in probiotic LAB strains are still absent and glutathione synthesis in them has not been demonstrated.

Effects of a synbiotic product on blood antioxidative activity in subjects colonized with Helicobacter pylori.

AIM: To evaluate the impact of the consumption of a synbiotic product on the antioxidative activity markers of blood in asymptomatic H. pylori-colonized persons. METHODS AND RESULTS: Fifty-three healthy adult volunteers without gastric symptoms participated in a randomized, double-blind placebo-controlled study. The crossover consumption of the enterocoated capsules containing antioxidative Lactobacillusfermentum ME-3, Lact. paracasei 8700:2 and Bifidobacterium longum 46 with Raftilose P95 lasted for 3 weeks and did not change the H. pylori colonization. In H. pylori-positive subjects the sera values of total antioxidative status (TAS) were significantly lower compared to H. pylori-negative subjects (0.97 vs 1.05 mmol l(-1), P = 0.008). After the consumption of the synbiotic, TAS values (0.97 vs 1.03 mmol l(-1), P = 0.004) increased, while the ratio between oxidized and reduced glutathione (0.035 vs 0.030, P = 0.016) decreased in H. pylori-positive subjects. CONCLUSIONS: The consumption of a synbiotic containing an antioxidative probiotic strain improved the reduced systemic antioxidative activity in H. pylori-colonized asymptomatic subjects. SIGNIFICANCE AND IMPACT OF THE STUDY: A synbiotic product containing an antioxidative probiotic strain may be useful in the reduction of systemic oxidative stress in H. pylori infection.

Characterization of oral lactobacilli as potential probiotics for oral health.

INTRODUCTION: Intestinal lactobacilli have been successfully used as probiotics to treat gastrointestinal disorders, but only limited data are available for the probiotic properties of oral lactobacilli to combat oral diseases. We aimed to characterize oral lactobacilli for their potential probiotic properties according to the international guidelines for the evaluation of probiotics, and to select potential probiotic strains for oral health. METHODS: The study included 67 salivary and subgingival lactobacilli of 10 species, isolated from healthy humans. All strains were identified using amplified ribosomal DNA restriction analysis, tested for antimicrobial activity against oral pathogens, tolerance of low pH and bile content. Thereafter, the lysozyme tolerance and antibiotic susceptibility of 22 potential probiotic strains were assessed. RESULTS: The majority of strains suppressed the growth of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, and Streptococcus mutans, but none inhibited Candida albicans. The lowest pH tolerated by lactobacilli following 4 h of incubation was pH 2.5, but none of the strains grew at this pH. All strains tolerated a high concentration of lysozyme (10 mg/ml) and half of the strains tolerated a high concentration of human bile [5% volume/volume (V/V)]. Four Lactobacillus plantarum and two Lactobacillus oris strains expressed resistance to tetracycline and/or doxycycline. CONCLUSIONS: Strains of L. plantarum, Lactobacillus paracasei, Lactobacillus salivarius, and Lactobacillus rhamnosus expressed both high antimicrobial activity and high tolerance of environmental stress. The absence of transferable antibiotic-resistance genes in L. plantarum strains remains to be confirmed. These results suggest a potential for oral lactobacilli to be used as probiotics for oral health.

Antagonistic activity of probiotic lactobacilli and bifidobacteria against entero- and uropathogens.

AIM: To develop in vitro assays for comparing the antagonistic properties and anti-oxidative activity of probiotic Lactobacillus and Bifidobacterium strains against various entero- and urinary pathogens. METHODS AND RESULTS: The antagonistic activity of five probiotic lactobacilli (Lactobacillus rhamnosus GG, Lactobacillus fermentum ME-3, Lactobacillus acidophilus La5, Lactobacillus plantarum 299v and Lactobacillus paracasei 8700:2) and two bifidobacteria (Bifidobacterium lactis Bb12, Bifidobacterium longum 46) against six target pathogens was estimated using different assays (solid and liquid media, anaerobic and microaerobic cultivation) and ranked (low, intermediate and high). Bacterial fermentation products were determined by gas chromatography, and the total anti-oxidative activity of probiotics was measured using linolenic acid test. Pyelonephritic Escherichia coli was highly suppressed by GG and both bifidobacteria strains. Lactobacilli strains 8700:2, 299v and ME-3 were the most effective against Salmonella enterica ssp. enterica in microaerobic while ME-3 and both bifidobacteria expressed high activity against Shigella sonnei in anaerobic milieu. Lact. paracasei, Lact. rhamnosus and Lact. plantarum strains showed intermediate antagonistic activity against Helicobacter pylori under microaerobic conditions on solid media. The highest anti-oxidative activity was characteristic for Lact. fermentum ME-3 (P < 0.05). No efficient antagonist against Clostridium difficile was found. The positive correlations between the pH, lactic acid production and anti-microbial activity for all tested probiotics were assessed. CONCLUSIONS: Developed experimental assays enable to compare the anti-microbial and -oxidative activity of Lactobacillus and/or Bifidobacterium probiotics, which have been claimed to possess the ability of suppressing the growth of various enteric and urinary pathogens. SIGNIFICANCE AND IMPACT OF THE STUDY: Screening Lactobacillus and Bifidobacterium sp. strains according to their activity in various environmental conditions could precede the clinical efficacy studies for adjunct treatment with probiotics in cure of different gastrointestinal and urinary tract infections.

Expression of single-chain antibody against RgpA protease of Porphyromonas gingivalis in Lactobacillus.

AIMS: The monoclonal antibody 61BG1.3, recognizing the RgpA protease, has been reported to confer protection against recolonization by the periodontal pathogen Porphyromonas gingivalis in humans. The aim of this study was to express a functional scFv derived from the monoclonal antibody 61BG1.3 on the surface of Lactobacillus paracasei for potential use in the prevention or treatment of periodontal diseases. METHODS AND RESULTS: The scFv was fused to an E-tag and cloned in the Escherischia coli/Lactobacillus shuttle vector pLP501, which mediates surface expression of the scFv. FACS analysis using an anti-E-tag antibody revealed that the scFv was expressed on the surface of the transformed lactobacilli and binding of the scFv to RgpA was shown by ELISA. Lact. paracasei expressing the scFv against RgpA was able to agglutinate P. gingivalis whereas the Lact. paracasei expressing an irrelevant scFv fragment did not. Scanning electron microscopy demonstrated efficient binding of the lactobacilli expressing the scFv anti-RgpA to P. gingivalis. CONCLUSIONS: We have expressed a functional scFv antibody directed against the RgpA protease of P. gingivalis in Lactobacillus. SIGNIFICANCE AND IMPACT OF THE STUDY: These results suggest a potential of Lactobacillus expressing scFvs against P. gingivalis to be used to combat periodontal disease.

Oral lactobacilli in chronic periodontitis and periodontal health: species composition and antimicrobial activity.

BACKGROUND/AIMS: Lactobacilli are known to play an important role in the maintenance of health by stimulating natural immunity and contributing to the balance of microflora. However, their role in chronic periodontitis is unclear. We aimed to identify oral lactobacilli in chronic periodontitis and periodontally healthy subjects, and to determine their antimicrobial activity against putative oral pathogens. METHODS: A total of 238 Lactobacillus isolates from the saliva and subgingival sites of 20 chronic periodontitis and 15 healthy subjects were collected. In all, 115 strains were identified using rapid amplified ribosomal DNA restriction analysis. Antimicrobial activity against Streptococcus mutans, Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Prevotella intermedia was assessed. RESULTS: Lactobacilli belonging to 10 species were identified. The most prevalent strains in healthy persons were Lactobacillus gasseri and Lactobacillus fermentum and in chronic periodontitis patients, Lactobacillus plantarum. Obligately homofermentatives, particularly L. gasseri, were less prevalent in chronic periodontitis patients compared with healthy subjects (8% vs. 64% for L. gasseri, P < 0.01). Sixty-nine percent of tested lactobacilli inhibited S. mutans, 88% A. actinomycetemcomitans, 82% P. gingivalis and 65% P. intermedia. The strongest antimicrobial activity was associated with Lactobacillus paracasei, L. plantarum, Lactobacillus rhamnosus, and Lactobacillus salivarius. The strains from periodontally healthy patients showed a lower antimicrobial activity against S. mutans than the strains from chronic periodontitis patients. CONCLUSION: The composition of oral lactoflora in chronic periodontitis and healthy subjects differs, with a higher prevalence of homofermentative lactobacilli, particularly L. gasseri, in the latter group. Both homo- and heterofermentative oral lactobacilli suppress the growth of periodontal pathogens, but the antimicrobial properties are strain, species and origin specific.

Intestinal microbiota and immunoglobulin E responses in 5-year-old Estonian children.

BACKGROUND: Over the last few decades, several studies from different parts of the world have indicated an increasing prevalence of allergic diseases. This has been related to environmental factors, like changes of microbial pressure. Our previous studies have demonstrated differences in the intestinal microbiota between allergic and non-allergic children. AIM: To test the hypothesis that the intestinal microbiota and IgE response are related, both in allergic and non-allergic 5-year-old Estonian children. METHODS: The study group comprised 19 allergic and 19 non-allergic 5-year-old children, selected from a larger group who had been followed from birth. The diagnosis of allergy was based on clinical examination of the children and on data obtained from the questionnaires. The faecal microbiota were quantified by seeding serial dilutions on nine different media for incubation in different environment. The composition of the gut microbiota was expressed both as absolute counts of the various species and their relative share among the total counts of identified microbiota. RESULTS: Bifidobacteria were less commonly detected in children with allergic diseases than in healthy children and clostridia comprised a higher proportion among their gut microbes. Children with specific IgE antibodies to defined allergens had higher counts of clostridia and the counts of clostridia correlated with the level of serum IgE, but only so in allergic children. In non-allergic children, the serum IgE levels showed a positive correlation with the counts of bacteroides. CONCLUSION: The development of allergic diseases seems to be associated with the composition of the gut microbial ecosystem. High counts of potential pathogens, such as clostridia, are associated with clinical manifestations of allergy and IgE antibody formation.

A new probiotic cheese with antioxidative and antimicrobial activity.

The aim of our study was to develop an original probiotic cheese based on the Estonian open-texture, smear-ripened, semisoft cheese "Pikantne." Cheese was produced by two methods using cheese starter cultures (Probat 505) in combination with 0.04% of probiotic Lactobacillus fermentum strain ME-3 (10(9) cfu/mL) with high antimicrobial activity and antioxidative properties. The probiotic Lactobacillus was added into milk simultaneously with starter cultures (cheese A) and into drained curd (cheese B). After addition of probiotic L. fermentum ME-3, the cheese composition, flavor, and aroma were comparable to the control cheese (score values = 4.5, 4.2, and 3.7 for control cheese, cheese A, and cheese B, respectively). Cheese A, which had good sensory properties, was chosen for further testing of viability and probiotic properties. The probiotic strain was found to withstand the technological processing of cheese, surviving and sustaining moderate antimicrobial and high antioxidative activity throughout ripening and storage (the ripened cheese contained approximately 5 x 10(7) cfu/g viable ME-3 cells), although the viability of the ME-3 strain incorporated into the cheese showed a slight decrease between d 24 and 54 after cheese preparation. Semisoft cheese "Pikantne" serves as a suitable carrier of antimicrobial and antioxidative L. fermentum ME-3.

Characterization of intestinal lactobacilli as putative probiotic candidates.

AIMS: To use antioxidative activity and antagonistic properties of lactobacilli against selected pathogens and members of the normal microflora as a basis for screening probiotic candidates. METHODS AND RESULTS: Antagonistic activity of lactobacilli against target bacteria in both microaerobic and anaerobic environments was tested. Production of antagonistic metabolites (ethanol, hydrogen peroxide (H2O2), acetic, lactic and succinic acid) by lactobacilli as well as their total antioxidative activity were assessed. In general, the lactobacilli tested were most effective against Gram-negative bacteria and their antagonistic activity was strain-specific. However, obligately heterofermentative lactobacilli had the strongest activity when tested in a microaerobic environment. Additionally, facultatively heterofermentative lactobacilli were equally effective in either milieu and produced significant levels of acetic and lactic acid. Moreover, obligately homofermentative lactobacilli had high H2O2 production and total antioxidative activity but weak antagonistic activity. CONCLUSIONS: Antioxidative and antagonistic activity of intestinal lactobacilli is strain-specific but typically can be related to their fermentation type which may be used for rapidly screening large numbers of lactobacilli for probiotic candidates. SIGNIFICANCE AND IMPACT OF THE STUDY: This study represents the first report on the utilization of group characteristics to screen lactobacilli intended for specific probiotic use. Such uses include the targeting of particular gut niches and pathogens as well as allowing for long-term benefits to the host.

Prevalence of antibiotic resistance of Helicobacter pylori isolates in Estonia during 1995-2000 in comparison to the consumption of antibiotics used in treatment regimens.

OBJECTIVE: To find a possible relation between the dynamics of antibiotic resistance of Helicobacter pylori isolates and the consumption of antibiotics during the last several years in Estonia. METHODS: Helicobacter pylori isolates were collected from the gastric mucosa of patients with peptic ulcer (153) and gastritis (68) and isolated on the Columbia Agar Base. From 1995 to 1997 the disk-diffusion method was used for testing of H. pylori susceptibility to metronidazole (115 isolates), erythromycin (119 isolates), tetracycline (119 isolates) and amoxicillin (119 isolates). From 1998 to 2000 the susceptibility of H. pylori to metronidazole (106 isolates), amoxicillin (30 isolates), clarithromycin (106 isolates) and ciprofloxacin (30 isolates) was assessed by E tests. Data from the Estonian State Agency of Medicines were used to determine the antibiotic consumption rate. RESULTS: Up to the year 2000 all the investigated H. pylori isolates were susceptible to ciprofloxacin; the resistance to clarithromycin, tetracycline, amoxicillin and erythromycin was 3%, 1.7%, 0.7% and 2.5%, respectively. Forty-six percent of H. pylori isolates were resistant to metronidazole. During 1995-2000 the consumption of amoxicillin, erythromycin and ciprofloxacin increased and the consumption of tetracycline decreased. The increasing consumption of amoxicillin reached a level 5.7 times than that of the consistent use of metronidazole. The resistance to amoxicillin appeared to be very low and resistance to metronidazole was continuously high. The increase of clarithromycin consumption (from 0.002 to 1.119 defined daily doses/1000) during three years was associated with the appearance of the first clarithromycin-resistant isolates in 2000. CONCLUSION: No relation was observed between the antibiotic consumption rate and the resistance pattern of H. pylori to metronidazole, amoxicillin, erythromycin, tetracycline and ciprofloxacin during recent years among the in population.

Antibiotic resistance as an indicator of bacterial chlorhexidine susceptibility.

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.

Allergy development and the intestinal microflora during the first year of life.

BACKGROUND: The intestinal microflora is a likely source for the induction of immune deviation in infancy. OBJECTIVE: The purpose of this study was to prospectively relate the intestinal microflora to allergy development in 2 countries differing with respect to the prevalence of atopic diseases. METHODS: Newborn infants were followed prospectively through the first 2 years of life in Estonia (n = 24) and Sweden (n = 20). By that age, 9 Estonian and 9 Swedish infants had developed atopic dermatitis and/or positive skin prick test results. Stool samples were obtained at 5 to 6 days and at 1, 3, 6, and 12 months, and 13 groups of aerobic and anaerobic microorganisms were cultivated through use of standard methods. RESULTS: In comparison with healthy infants, babies who developed allergy were less often colonized with enterococci during the first month of life (72% vs 96%; P <.05) and with bifidobacteria during the first year of life (17% to 39% vs 42% to 69%; P <.05). Furthermore, allergic infants had higher counts of clostridia at 3 months (median value, 10.3 vs 7.2 log(10); P <.05). The prevalence of colonization with Staphylococcus aureus was also higher at 6 months (61% vs 23%; P <.05), whereas the counts of Bacteroides were lower at 12 months (9.9 vs 10.6 log(10); P <.05). CONCLUSION: Differences in the composition of the gut flora between infants who will and infants who will not develop allergy are demonstrable before the development of any clinical manifestations of atopy. Because the observations were made in 2 countries with different standards of living, we believe that our findings could indicate a role for the intestinal microflora in the development of and protection from allergy.

Antibiotic susceptibility patterns of community- and hospital-acquired Staphylococcus aureus and Escherichia coli in Estonia.

This study compares the susceptibility patterns of Staphylococcus aureus and Escherichia coli isolated from patients with hospital-acquired and outpatient infections. A total of 902 isolates of S. aureus and 1,114 of E. coli were collected in five different Estonian medical centers between January 1997 and November 1997. Strains were grouped into two different categories, depending on whether they had been obtained from inpatients or outpatients. Compared to S. aureus strains isolated from inpatients, the strains from outpatients were significantly more resistant to erythromycin (25.3% vs. 17.9%), tetracycline (33.5% vs. 22.4%) and trimethoprim-sulfamethoxazole (13.9% vs. 7.9%). The overall prevalence of oxacillin-resistant S. aureus was 10.4%, with no significant differences noted between isolates recovered from inpatients and outpatients. In the case of E. coli, significantly more isolates from inpatients (42.8%) than from outpatients (34.4%) were ampicillin-resistant. Inpatient isolates of E. coli were also more resistant to cefotaxime (9.3%) and nitrofurantoin (11.2%) than outpatient strains (0% and 3.1%, respectively). Analysis showed remarkable co-resistance among both inpatient and outpatient strains of S. aureus and E. coli. Multiple resistant S. aureus and E. coli strains represented 15.1% and 17.3%, respectively of the organisms examined in this study. With respect to E. coli, significantly more multiresistant isolates were found in inpatient than outpatient isolates (20.4% vs. 8.9%). Our results indicate that the distinction between community-acquired and hospital infections due to S. aureus and E. coli may not be valid in Estonia.