Detection of Chlamydophila Pneumoniae and Typical Bacteria in Patients with Chronic Cough.

The aim of research was to analyze the results of microbiological tests for typical and atypical bacteria in patients with symptoms of chronic cough. A total of 214 outpatients aged from 2 to 94 years (110 women, 64 men, and 40 children) with chronic cough were studied. Four hundred twenty eight throat swabs were examined for atypical bacteria antigen (Chlamydophila pneumoniae) (n=214) and typical pathogens (n=214). Chl. pneumoniae detection was performed using indirect immunofluorescence test. Classical microbiological culture was used for typical bacteria detection. Chl. pneumoniae antigen was detected in 55/214 (26.0%) patients with chronic cough (in 31 (28.2%) women, 14 (21.9%) men, and 10 (25.0%) children). Positive culture for typical pathogens was observed in 30 (27.3%) women, 22 (34.4%) men, and 21 (52.5%) children. Simultaneous occurrence of Chl. pneumoniae and typical pathogens (Staphylococcus aureus strain MSSA, Streptococcus pyogenes, or Moraxella catarrhalis) was found in 16 (7.5%) patients. The findings show that in patients with chronic cough Chl. pneumoniae infection, although less than that with typical pathogens, is rather frequent. Further, the performance of test for Chl. pneumoniae in throat swabs from patients with chronic cough is good and provides an efficient way to diagnose the infection and implement appropriate therapy.

Genotypic and clinical differences of seropositive Helicobacter pylori children and adults in the Polish population.

Helicobacter pylori (H. pylori) plays an important role in the pathogenesis of the upper gastrointestinal tract diseases in both children and adults. The aim of this paper was to assess the differences between the clinical course of the disease in children and adults. This paper also presents an analysis of clinical symptoms, endoscopic and histopathological findings, H. Pylori cagA and vacA genotypes rates and analysis of the sensitivity of these strains to antibiotics in the Polish population, with possible practical and therapeutic implications. The multicenter study on the frequency of H. pylori infections assessed by the presence of antibodies in IgG class against H. pylori in serum was conducted in the years 2002 and 2003. The study group included 6565 children and adults, in 3827 of whom antibodies levels were above 24 U/mL. The authors analyzed clinical and endoscopic symptoms and in some patients with H. pylori seropositivity also histopathological changes, and cagA and vacA genes. Sensitivity of H. pylori strains to antibiotics were also analyzed. Differences between the frequency of infection between children and adults were determined. Endoscopic examination in adults revealed more frequent cases of gastropathy (P=0.003) and erosive gastritis (P=0.001), and in children-thick mucosal folds (P<0.0001). Histopathological examinations carried out in adults have revealed atrophic gastritis and intestinal metaplasia. In children, cagA(+) s1m1 was observed more frequently than in adults (34.0% versus 23.1%; P=0.02) contrary to cagA(-)s2m2 which occurred more frequently in adults (27.1% versus 14.0%; P=0.003). No effect of the infection on nausea, regurgitation, vomiting, heartburn, and abdominal pain in children was detected. However, adults infected with H. pylori suffered from more frequent episodes of heartburn and abdominal pain. The H. pylori strain exhibited a high resistance to metronidazole (higher in adults: 41.7% versus 27.4%; P=0.002), and to clarithromycin (higher in children: 20.2% versus 15.4%; P>0.05), and dual resistance to metronidazole and clarithromycin (higher in children: 9.9% versus 8.4%; P>0.05). Resistance of the H. pylori to amoxicillin and tetracycline was not detected. The conducted study indicated clinical differences in the H. pylori infection in children and adults. Among the differences in children, especially the more frequent infections by the cagA(+)s1m1/m2 strain could have an influence on further consequences of the infection. The obtained results could be useful in therapeutic decisions.

The clinical aspects of Helicobacter heilmannii infection in children with dyspeptic symptoms.

UNLABELLED: Helicobacter heilmannii (H. heilmannii) infection is a relatively rare causative agent of gastroduodenal diseases in children. However, H. heilmannii frequently colonizes gastric mucosa of animals, mainly cats and dogs, from where it can be transmitted to humans. The aim of the study was to evaluate the incidence of H. heilmannii infection in children with dyspeptic symptoms treated in our clinic. A number of 13,124 esophagogastroduodenoscopies in children aged 4 to 18 years were conducted from 1992 to 2010. The indications for examination were: chronic abdominal pain, nausea, vomiting, heartburn, anaemia, disturbances of intestinal absorption and other. In 11,023 cases microbiologic studies and cultures toward Helicobacter infection were carried out and in 22 children H. heilmannii infection was confirmed. H. heilmannii infection was diagnosed based on morphologic examination in direct microscopy of biopsy specimens from gastric mucosa. In children with H. heilmannii infection clinical symptoms, contact with animals, endoscopic findings of the upper gastrointestinal tract and results of diagnostic tests for Helicobacter pylori infection were assessed. In our studies H. heilmannii infection was diagnosed in 22 children. The rate of H. heilmannii infection was 0.2% in examination of gastric mucosa specimens. No sex-dependent difference in the rates was observed. Most of the children lived in cities and 54.5% had contact with dogs and/or cats. Children complained of chronic epigastric pain, nausea, vomiting and heartburn. Endoscopic studies most often revealed nodular gastritis and gastric or duodenal ulcer in two children. In three children result of the endoscopic study was normal. CONCLUSIONS: H. heilmannii infection in children is rare. However, it may be one of the causes of gastroduodenal diseases in children.

Resistance patterns and occurrence of virulence determinants among GRE strains in southwestern Poland.

PURPOSE: The aim of this study was to determine the antimicrobial resistance and the occurrence of virulence determinants among glycopeptide-resistant enterococci (GRE) isolated in 2007-2009 from patients hospitalized in southwestern Poland. MATERIAL AND METHODS: The minimal inhibitory concentrations (MICs) of antibiotics were determined by agar dilution method or by E-test®. The presence of vanA - vanG resistance and virulence genes (agg, esp, gelE and cylA, cylB, cylM) was investigated using PCR. The ability to form biofilm and the activity of gelatinase, hemolysins, lipase and DNase were tested. RESULTS: All the GRE strains were susceptible to linezolid, daptomycin, and tigecycline and resistant to norfloxacin. In the Enterococcus faecium group, 17 strains carried the vanA gene and 20 the vanB gene. In the Enterococcu faecalis group, 4 strains carried the vanA gene and 1 the vanB gene. There were differences in tetracycline susceptibility between the VanA (70%) and the VanB (55%) phenotypes. Only linezolid had high activity against both the VanA and the VanB phenotypes. The esp gene was present in most of the GRE strains, but only 3 E. faecalis strains produced biofilm. Lipase was produced by 10/42 examined strains, gelatinase by 4/42 and hemolysin by 3/42 isolates. CONCLUSIONS: Linezolid seems to be the optimal option in empirical therapy of infections caused by GRE strains because of the relationship between its activity (MIC value) and susceptibility breakpoint. There was no correlation between the prevalence of different virulence genes and resistance to the antibiotics tested.

Respiratory infection caused by Chlamydophila pneumoniae in children and adolescents in the Lower Silesia Region of Poland.

OBJECTIVE: To assess the incidence of Chlamydia pneumoniae respiratory tract infection in children and adolescents in the Lower Silesia Region in Poland in 2009. MATERIAL AND METHODS: 641 throat swabs obtained from 326 girls and 315 boys, aged 11 months to 18 years, were assessed diagnostically. The patients enrolled into the study were treated on an outpatient basis due to various, non-specific respiratory ailments. The most common presenting clinical symptom of a respiratory problem was dry cough, which occurred in 295 studied subjects, followed by runny nose and cough with discharge in 176 subjects, and other minor symptoms in 35 subjects. The assessment was conducted by an indirect immunofluorescence antibody (IFA) Chlamydia Testing kit (Cellabs, Sydney, Australia). RESULTS: Overall, Chlamydia infection was detected in the respiratory tract in 43.1% (276/ 641) of the children, with no clear gender differences. Of the 295 subjects presenting with dry cough, 122 (41.4%) had positive tests for Chlamydia. Of the 176 subjects with runny nose and cough and the 35 subjects with other symptoms, 83 (47.2%) and 8 (22.9%) had positive tests for Chlamydia, respectively. In the asymptomatic children who had direct contact with a Chlamydia infected person, there were 29.6% (8/27) positively tested cases, whereas in the children presenting symptoms, the percentage of positive tests was 48.3% (29/60). CONCLUSIONS: In children living in the Lower Silesia Region of Poland, there is a substantial ∼50% rate of Chlamydia infection, transmitted via airborne droplets. The finding of Chlamydia infection should be the signal for testing other subjects from the child's closest environment.

High level of resistance to metronidazole and clarithromycin in Helicobacter pylori isolated from pediatric patients in Poland (1997-2001).

Resistance to metronidazole (Met), clarithromycin (Cla) and amoxycillin (Amo) was examined using H. pylori isolates from child patients before and after treatment in the period 1997-2001. The rate of Met and Cla resistance before treatment was 35.2 and 8.6%, respectively. Six weeks after treatment 48.4% of the isolated strains were resistant to Met and 17.6% to Cla. The highest rate of resistance to both antibiotics was determined in 2001 (before treatment, 46 and 15%, respectively, and after treatment, 57.8 and 26.3%, respectively). All the strains were susceptible to Amo. Strains resistant to Met were detected more frequently in girls than in boys.

The status of antimicrobial resistance of Helicobacter pylori in eastern Europe.

OBJECTIVE: To evaluate the primary, secondary and combined resistance to five antimicrobial agents of 2340 Helicobacter pylori isolates from 19 centers in 10 countries in eastern Europe. METHODS: Data were available for centers in Bulgaria, Croatia, the Czech Republic, Estonia, Greece, Lithuania, Poland, Russia, Slovenia and Turkey. Susceptibility was tested by agar dilution (seven countries), E test (five countries) and disk diffusion (three countries) methods. Resistance breakpoints (mg/L) were: metronidazole 8, clarithromycin 1, amoxicillin 0.5, tetracycline 4, and ciprofloxacin 1 or 4 in most centers. Primary and post-treatment resistance was assessed in 2003 and 337 isolates respectively. Results for 282 children and 201 adults were compared. RESULTS: Primary resistance rates since 1998 were: metronidazole 37.9%, clarithromycin 9.5%, amoxicillin 0.9%, tetracycline 1.9%, ciprofloxacin 3.9%, and both metronidazole and clarithromycin 6.1%. Isolates from centers in Slovenia and Lithuania exhibited low resistance rates. Since 1998, amoxicillin resistance has been detected in the southeastern region. From 1996, metronidazole resistance increased significantly from 30.5% to 36.4%, while clarithromycin resistance increased slightly from 8.9% to 10.6%. In centers in Greece, Poland, and Bulgaria, the mean metronidazole resistance was slightly higher in adults than in children (39% versus 31.2%, P > 0.05); this trend was not found for clarithromycin or amoxicillin (P > 0.20). Post-treatment resistance rates exhibited wide variations. CONCLUSIONS: In eastern Europe, primary H. pylori resistance to metronidazole is considerable, and that to clarithromycin is similar to or slightly higher than that in western Europe. Resistance to amoxicillin, ciprofloxacin and tetracycline was detected in several centers. Primary and post-treatment resistance rates vary greatly between centers.

Neutralisation of cytotoxic vacuolating activity by serum antibodies of Helicobacter pylori-infected patients.

The study involved 196 H. pylori strains and 196 serum samples taken from the same patients. H. pylori strains were investigated for the production of vacuolating cytotoxin. Antibodies to the vacuolating cytotoxin produced by H. pylori were detected in the sera samples by neutralisation assay (on Intestine 407 cells) and ELISA. Of the 196 H. pylori strains tested, 80 (40.8%) were found to express vacuolating cytotoxic activity. The titres of cytotoxic nonconcentrated broth culture filtrates ranged from 1:2 to 1:128. The vacuolating assay was positive in 37.1% strains isolated from children, and in 50% strains isolated from adults. Cytotoxin-positive H. pylori strains were found more frequently in duodenal ulcer (71%) than in chronic gastritis (35.2%) patients, and this difference was statistically significant p < 0.05. Neutralising antibodies to vacuolating cytotoxin were present in 51% and 49% of the serum samples tested by neutralisation and ELISA, respectively. Duodenal ulcer patients had antibodies to vacuolating cytotoxin more frequently (p < 0.05) than chronic gastritis patients. Antibodies to cytotoxin were detected in the serum samples from patients infected by cytotoxic (100%) and noncytotoxic (18%) H. pylori strains.

Identification of a putative chromosomal replication origin from Helicobacter pylori and its interaction with the initiator protein DnaA.

The key elements of the initiation of Helicobacter pylori chromosome replication, DnaA protein and putative oriC region, have been characterized. The gene arrangement in the H.pylori dnaA region differs from that found in many other eubacterial dnaA regions (rnpA-rmpH-dnaA-dnaN-recF-gyrB). Helicobacter pylori dnaA is flanked by two open reading frames with unknown function, while dnaN-gyrB and rnpA-rmpH loci are separated from the dnaA gene by 600 and 90 kb, respectively. We show that the dnaA gene encoding initiator protein DnaA is expressed in H.pylori cells. The H.pylori DnaA protein, like other DnaA proteins, can be divided into four domains. Here we demonstrate that the C-terminal domain of H.pylori DnaA protein is responsible for DNA binding. Using in silico and in vitro studies, the putative oriC region containing five DnaA boxes has been located upstream of the dnaA gene. DNase I and gel retardation analyses show that the C-terminal domain of H.pylori DnaA protein specifically binds each of five DnaA boxes.

Neutralisation of vacuolating activity of cytotoxin by serum antibodies of Helicobacter pylori infected patients.

The study involved 196 H. pylori strains and 196 serum samples taken from the same patients. H. pylori strains were investigated for the production of vacuolating cytotoxin. Antibodies to the vacuolating cytotoxin produced by H. pylori were detected in the sera samples by neutralisation assay (on Intestine 407 cells) and ELISA. Of the 196 H. pylori strains tested, 80 (40.8%) were found to express vacuolating cytotoxic activity. The titres of vacuolating cytotoxic were ranged from 1:2 to 1:128. The vacuolating assay was positive in 37.1% strains isolated from children, and in 50% strains isolated from adults. Cytotoxin-positive H. pylori strains were found more frequently in duodenal ulcer (71%) than in chronic gastritis (35.2%) patients, and this difference was statistically significant p<0.05. Neutralising antibodies to vacuolating cytotoxin were present in 51% and 49% of the serum samples tested by neutralisation and ELISA, respectively. Duodenal ulcer patients had antibodies to vacuolating cytotoxin more frequently (p<0.05) than chronic gastritis patients. Antibodies to cytotoxin were detected in 100% of the serum samples from patients infected by cytotoxic H. pylori strains. This suggests that the presence of anticytotoxic antibodies in the serum samples may be regarded as a sensitive indicator of infection by cytotoxic H. pylori strains.

[Characteristic of verotoxigenic strains of Escherichia coli]. / Charakterystyka verotoksycznych szczepów Escherichia coli.

The aim of the study was the isolation from faecal samples of patients with diarrhoea of verotoxigenic strains of E. coli (VTEC) on the basis of characteristic biochemical properties and production of enterohaemolysin and comparison of isolated verotoxigenic strains with reference strains of VTEC. For isolation of VTEC from 257 stool samples derived from patients with diarrhoea were used selective medium sorbiol--Mac Conkey agar (SMAC) and media supplemented with unwashed and washed in PBS sheep erythrocytes for detection of haemolysins of E. coli. In all haemolytic and sorbitolo-positive or -negative strains isolated from 93 stool samples were examined the activity of beta-glucuronidase using MUG (4-methylumbelliferyl-beta-D-glukuronid) as a substrate for that enzyme. All isolated haemolytic strains as well as reference VTEC were examined on Vero cell line. Verotoxigenic strains from examined samples were investigated by agglutination assay with antiserum to E. coli O157 and then with antisera to eneropathogenic E. coli (EPEC). After that they were examined with ID GN and ATB GN tests. In 93 (36.2%) examined samples there were haemolytic strains of E. coli which fermented or not sorbitol and were MUG-positive or negative. Only in 2 (0.2%) stool samples there were verotoxigenic strains of E. coli which were sorbiol-positive and MG-positive. Both strains belonged to O26 serotype and were derived from samples of two children with diarrhoea. Isolated verotoxigenic strains of E. coli O26 were susceptible on all tested antibiotics.

[Frequency of Helicobacter pylori infection depending on morphological changes in gastric mucosa in patients over 65 years of age]. / Czestosc zakazenia Helicobacter pylori w zaleznosci od zmian morfologicznych blony sluzowej zoladka u ludzi po 65 roku zycia.

In older patients involution processes are observed. Frequency of H. pylori infection has been related of histopathological changes of gastric mucosa in patients over 65 yrs old. In 73% of analyzed patients gastritis atrophica has been documented.

Relationship between antibody to cytotoxin and Helicobacter pylori infection.

Broth culture supernatants from 14 (34%) out of the 41 H. pylori strains tested, induced vacuolization in Intestine 407 cells in titers ranging from 1:2 to 1:64. 20% of H. pylori strains isolated from children and 42% of strains isolated from adults expressed vacuolating activity. Serum antibody to cytotoxin produced by H. pylori was detected with a neutralization assay. Anticytotoxic antibodies were present in all sera from patients infected with cytotoxic H. pylori strains. The toxin-neutralizing activity of sera from individuals infected with H. pylori suggests that the cytotoxin is produced in vivo.

Helicobacter pylori in kidney allograft recipients: high prevalence of colonization and low incidence of active inflammatory lesions.

Since kidney transplant recipients are at enhanced risk for developing severe upper gastrointestinal disease and Helicobacter pylori (Hp) is an important pathogen in active gastritis and peptic ulcer, we performed gastroduodenoscopic examination, coupled with assessment of Hp colonization in 29 renal allograft recipients complaining of recurrent dyspepsia. Results were compared with those of 25 chronically hemodialyzed patients and 16 subjects free from renal disease, also suffering from upper gastrointestinal symptoms of similar severity. We found that while transplant recipients have had a high prevalence of Hp infection (62 vs. 34.6% in dialysis and 43.6% in control dyspeptic patients), active gastritis was clearly less frequently seen in these patients than in control subjects (transplant group: 6.9%, dialysis 3.8%, control 31.3%) and peptic ulceration was totally absent. Prevalence of Hp colonization was even higher in renal graft recipients on triple posttransplant immunosuppression (82%). In dyspeptic transplant and dialysis patients, colonization with Hp did not account for development of active inflammatory lesions, an association frequently seen in subjects free from renal disease and immunosuppressive therapy.

IgG and IgA antibodies in Helicobacter pylori infections.

We compared Helicobacter pylori-specific IgG and IgA antibodies in serum, salivary and gastric juice of H. pylori-infected and noninfected patients using the ELISA method. Serum and salivary anti-H. pylori IgG and IgA titres were higher in H. pylori-positive than H. pylori-negative patients. Serum H. pylori IgG titres were higher than those of serum-specific IgA. In contrast, H. pylori IgA titres in saliva samples were higher than those of specific IgG. In all patients, the levels of IgG antibodies were very low in gastric juice. H. pylori-positive patients with increased levels of IgA in sera, also exhibited increased IgA levels in gastric juice. The determination of IgA antibodies in serum and saliva may be used as a test that complements the IgG antibody assay.

Helicobacter pylori infection in Eastern Europe: seroprevalence in the Polish population of Lower Silesia.

The Helicobacter pylori status of the population of Eastern European countries has not been explored despite the high incidence of peptic ulcer disease and gastric cancer observed in these countries. A seroprevalence study has been performed in Wroclaw, a city of Lower Silesia, Poland, to provide insight into this question. Sera were collected to obtain 50 subjects per 5 yr increment of age. A second generation ELISA kit with a high sensitivity and specificity was used. The results plotted by year of birth show a very high prevalence of H. pylori infection in all adults groups born before 1970 (80-100% positive). In the younger age groups, a dramatic decrease was observed. Because it is now known that most H. pylori infections are acquired in childhood (cohort effect), it can be predicted that the infection rate in the adult population will be much lower in the future compared with that presently observed, and it can be expected that evolution in H. pylori prevalence will have an impact on the rate of gastroduodenal diseases in Poland. Because of the high prevalence, it was not possible to identify risk factors for infection in this population.

[Extraintestinal salmonellosis in patients with blood diseases caused by Salmonella enteritidis]. / Pozajelitowa salmoneloza u osób z chorobami krwi, wywolana przez Salmonella enteritidis.

Four cases of extraintestinal salmonellosis caused by Salmonella enteritidis were described. Underlying diseases in the three patients were haematological neoplasms (2 splenectomized and 1 with massive leukemic infiltrations of the spleen) and in the fourth haemophilia B: only that patient had a prior symptomatic intestinal infection. Blood cultures for S.ent. were positive in all patients and additionally in that suffering from Hodgkin's disease urine, lymph node and stool cultures also showed S.ent. In the haemophiliac patient culture of suppurated hematoma was positive. All patients recovered from S. sepsis but three then died of their neoplastic diseases. The haemophiliac patient is in a good condition. Various disturbances in the immunological tests were observed.

Helicobacter pylori antibodies in sera of children suffering from chronic abdominal pain.

107 pediatric patients aged 9 to 18 with persistent gastric complaints were examined serologically and bacteriologically for Helicobacter pylori. Helicobacter was identified in 48 (45%) of individuals. 51 (48%) of children were found to be seropositive when H. pylori antibodies were detected by the ELISA; 56 (52%) when the passive haemagglutination test was used, and 41 (38%) in the latex agglutination test. 25% of culture-negative patients were found to be seropositive. The percentage of raised H. pylori antibody titres in the control (healthy subjects) varied from 20 to 27%, depending on the method applied.