RESUMO
The aim of the study was to detect anti-Helicobacter pylori activity of seven Lactobacillus delbrueckii subsp. bulgaricus (GLB) strains by four cell-free supernatant (CFS) types. Activity of non-neutralized and non-heat-treated (CFSs1), non-neutralized and heat-treated (CFSs2), pH neutralized, catalase-treated and non-heat-treated (CFSs3), or neutralized, catalase- and heat-treated (CFSs4) CFSs against 18 H. pylori strains (11 of which with antibiotic resistance) was evaluated. All GLB strains produced bacteriocin-like inhibitory substances (BLISs), the neutralized CFSs of two GLB strains inhibited >81% of test strains and those of four GLB strains were active against >71% of antibiotic resistant strains. Two H. pylori strains were BLIS resistant. The heating did not reduce the CFS activity. Briefly, all GLB strains evaluated produced heat-stable BLISs, although GLB and H. pylori strain susceptibility patterns exhibited differences. Bacteriocin-like inhibitory substance activity can be an advantage for the probiotic choice for H. pylori infection control. SIGNIFICANCE AND IMPACT OF THE STUDY: In this study, anti-Helicobacter pylori activity of seven Lactobacillus delbrueckii subsp. bulgaricus (GLB) strains was evaluated by four cell-free supernatant (CFS) types. The GLB strains produced heat-stable bacteriocin-like inhibitory substances (BLISs) with a strong anti-H. pylori activity and some neutralized, catalase- and heat-treated CFSs inhibited >83% of the test strains. Bacteriocin-like inhibitory substance production of GLB strains can render them valuable probiotics in the control of H. pylori infection.
Assuntos
Antibacterianos/farmacologia , Bacteriocinas/farmacologia , Helicobacter pylori/efeitos dos fármacos , Lactobacillus delbrueckii/metabolismo , Probióticos/farmacologia , Bacteriocinas/metabolismo , Catalase/metabolismo , Gastroenterite/microbiologia , Gastroenterite/terapia , Infecções por Helicobacter , Helicobacter pylori/isolamento & purificação , Humanos , Concentração de Íons de Hidrogênio , Lactobacillus delbrueckii/classificação , Lactobacillus delbrueckii/genética , Testes de Sensibilidade MicrobianaRESUMO
INTRODUCTION: Vulvovaginal candidiasis is common infection among those affecting the vulva and vagina. Is caused by the perpesentatives from the genus Candida, in most cases C. albicans (85-90%). An increase in the percentage of the so-called non-albicans agents is seen and these pathgogens are often resistant to the most commonly used in the practice antifungals. Faulty diagnosis, incorrect use of azoles, and self-treatment lead to selection of resistant strains and recurrent infections. AIM: Identification of Candida species associated with vulvovaginal candidiasis by conventional and PCR techniques. MATERIALS AND METHODS: For six months a total number of 213 vaginal secretions were tested applying Gram stain and cultivation on ChromAgar. API Candida fermentation tests and API 20CAUX assimilation tests were performed for the identification of the bacteria. Extraction of DNA of all the smears with subsequent PCR detection of different Candida species were done. RESULTS: 80.7% materials showed presence of blastospores and/or hyphae. Positive culture results were detected in 60 (28.2%) samples. The species specific identification revealed presence of C. albicans in 51 (85%) smears, C. glabrata--in 8 (13.3%), C. krusei--in 2 (3.3%), and S. cervisie--in 1 (2.1%). The PCR technique confirmed the results of the conventional methods. It is worth to mention that 51 of the tested smears were positive for G. vaginalis using additional PCR. CONCLUSIONS: The correct diagnosis of the cause of vulvovaginal candidiasis helps in the correct choice of appropriate antifungal therapy and prevents development of recurrent infections and consequences. The PCR based method is rapid, specific and sensitive. It perfectly correlates with the results from the conventional diagnostic tests so it could be selected as a method of choice for the diagnosis of vulvovaginal candidiasis.
Assuntos
Candida/isolamento & purificação , Candidíase Vulvovaginal/diagnóstico , Vagina/microbiologia , Candida/classificação , Candidíase Vulvovaginal/microbiologia , Feminino , Humanos , Reação em Cadeia da PolimeraseRESUMO
BACKGROUND: Bacterial vaginosis (BV) is the most common infection of the lower genital tract among women of reproductive age. The purpose of this study was to determine the frequency of BV and the important etiological agent Gardnerella vaginalis in Bulgarian patients of different age groups, as well as, the risk factors forBV METHODS: One thousand five hundred and twenty-three (1523) women aged 16 to 45 years without previous antimicrobial therapy were included in this study. The methods used were: scoring of Gram staining of vaginal smear and polymerase chain reaction (PCR) assay for G. vaginalis. RESULTS: Positive for BV according to the microscopic examination criteria were 47.80% from the 1523 tested women. In 88.87% from the positive samples G. vaginalis was detected using PCR, thus demonstrating high comparability of the both methods and the leading role of G. vaginalis. The most affected with BV is the age group of 21-25 years (33.21% of all subjects with BV). The most important detected risk factors were: smoking (more than 55% with BV), single marital status (only 15% with BV are married), more than I sexual partner (more than 36% had changed the sexual partner), early onset of sexual activity (75% of B V asocciated subjects started their sexual activity by the age 15-18 years). CONCLUSIONS: The established early age for aquiring BV among Bulgarian women is very important and alarming sign. This is the first study on the etiological role of G. vaginalis and on the risk factors for BV in Bulgaria.
Assuntos
Gardnerella vaginalis/isolamento & purificação , Infecções por Bactérias Gram-Positivas/epidemiologia , Vagina/microbiologia , Vaginose Bacteriana/epidemiologia , Adolescente , Adulto , Bulgária , Feminino , Violeta Genciana , Infecções por Bactérias Gram-Positivas/microbiologia , Humanos , Pessoa de Meia-Idade , Fenazinas , Fatores de Risco , Esfregaço Vaginal , Vaginose Bacteriana/microbiologia , Adulto JovemRESUMO
PURPOSE: The non-invasive pneumococcal disease (NIPD) is a common infection during childhood. We aimed to define the clonal spread of pediatric non-invasive isolates recovered during the PCV10-period in Bulgaria concerning the serotype and antimicrobial susceptibility. MATERIALS AND METHODS: Serogrouping/serotyping were performed using latex agglutination and capsular swelling reaction. Serogroup 6 strains were subjected to serotype-specific PCR's. The antibiotic susceptibilities were assessed by broth microdilution. MLST was performed to define the clonal composition. RESULTS: We analyzed 154 pediatrics non-invasive S. pneumoniae isolates. The PCV10-vaccinated children were 94.1%. We disclosed 88% non-vaccine serotypes (NVTs) and 12% PCV10 - serotypes. All common serotypes among PCV10-vaccinated children (n â= â145) were non-vaccine types (NVTs): 19A (13.8%), 6C (11.7%), 3 (9.6%), 15A (8.3%) and 23A (5.5%). Antimicrobial non-susceptibility showed highest levels in erythromycin (50.0%), oral penicillin (49.4%), clindamycin (45.4%), trimethoprim-sulfamethoxazole (43.5%), tetracycline (42.2%), and ceftriaxone (14.3%). The multidrug-resistant strains (MDR) were 51.3%. MDR-serotypes were 6C (20.2%), 19A (17.7%), 15A (11.4%), 19F (10.1%), and 23A (8.9%). MLST presented 17 clonal complexes (CCs) with prevalence of CC320, CC386, CC505, CC8029 and CC2613 clustered 83% MDR isolates. CONCLUSIONS: All emergent pediatric non-invasive serotypes in our geographic area during the studied PCV10-period were NVTs (19A, 6C, 3, 15A, and 23A). The fifth widespread CCs: CC320, CC386, CC505, CC8029 and CC2613 clustered 83% MDR isolates. Future surveillance of vaccine-induced changes in the clonality and the antimicrobial resistance of the pneumococcal population is needed.
Assuntos
Infecções Pneumocócicas , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Bulgária/epidemiologia , Criança , Humanos , Lactente , Testes de Sensibilidade Microbiana , Tipagem de Sequências Multilocus , Infecções Pneumocócicas/epidemiologia , Infecções Pneumocócicas/prevenção & controle , Vacinas Pneumocócicas , Sorogrupo , SorotipagemRESUMO
Chlamydia trachomatis infection is the most common sexually transmitted bacterial disease. The objective of this study was to establish the presence/absence of C. trachomatis in 98 patients with chronic complaints about the prostate and to evaluate the role of this bacterium in the inflammation of the gland. We performed culture and microscopical examination of pre-massage/post-massage urine and expressed prostatic secretions (EPS). In all cases, culture on McCoy cells and polymerase chain reaction (PCR) of the EPS was performed. Based on laboratory findings in 53 cases (54.08%), Escherichia coli, Klebsiella, Enterobacter, Proteus, Pseudomonas and Staphylococcus were isolated and accepted as causative agents of chronic bacterial prostatitis. Forty-five patients were categorised as patients with chronic pelvic pain syndrome. The results from the PCR and the cell culture for detection of C. trachomatis were as follows - two positive probes detected at the same time by applying PCR and cultivation and 1 positive only by PCR but not by cultivation on the cell line. Based on these results, it is concluded that C. trachomatis is not so frequently detected in our patients. C. trachomatis may be accepted as one of the aetiological agents of chronic prostatitis and testing for this infection is highly recommended when presumption for chronic prostatitis is apparent.
Assuntos
Infecções por Chlamydia/complicações , Chlamydia trachomatis/isolamento & purificação , Prostatite/microbiologia , Humanos , MasculinoRESUMO
AIMS: To evaluate the activities of six Lactobacillus delbrueckii subsp. bulgaricus (LB) strains against 30 Helicobacter pylori strains by agar-well diffusion method. METHODS AND RESULTS: LB cultures [4 x 10(8)-4 x 10(9) CFU ml(-1)) either were prepared in milk at their native pH, 3.8-5.0, or were adjusted to pH 6.4-7.7. At low and neutralized pH, LB strains inhibited the growth by 40-86.7% and 16.7-66.7% of H. pylori strains, respectively. LB activity was strain-dependent. At low and neutralized pH, one and five H. pylori strains, respectively, were not inhibited by any LB strain. LB2 and LB3, taken together, were active against most metronidazole and clarithromycin resistant strains. CONCLUSIONS: All LB strains inhibited a number of H. pylori strains, including also antibiotic resistant strains. LB activity was strain-dependent and better at low pH. At low pH values, the most active LB strains were LB1, LB2 and LB3, inhibiting 86.7% of H. pylori strains, while at neutralized pH values, the most active LB strains were LB2 and LB3, inhibiting 53.3 and 66.7% of H. pylori strains, respectively. SIGNIFICANCE AND IMPACT OF THE STUDY: LB could be utilized in the treatment or prophylaxis of H. pylori infection and warrants clinical investigations.
Assuntos
Antibiose , Infecções por Helicobacter/microbiologia , Helicobacter pylori/fisiologia , Lactobacillus/fisiologia , Adolescente , Adulto , Idoso , Antibacterianos/farmacologia , Criança , Feminino , Helicobacter pylori/efeitos dos fármacos , Helicobacter pylori/isolamento & purificação , Humanos , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Adulto JovemRESUMO
Chlamydia trachomatis infection is the most common bacterial sexually transmitted disease supposed to cause urethritis, epididymitis, prostatitis and infertility in men. The objective of this study was to assess the frequency of C. trachomatis infection in male partners of infertile couples at childbearing age. Sixty infertile couples and a control group of 40 healthy volunteers were included in the study. Urethral swabs were taken from all the male participants and cervical swabs from the female partners of the infertile couples. Culturing on McCoy cell line and PCR were the methods used for detection of the infection. C. trachomatis was found in five out of the 60 male urethral samples. Three of the female partners of these five positive males were diagnosed with C. trachomatis infection, too. We registered a woman with C. trachomatis infection whose partner's samples were negative for the bacterium. The control group showed one specimen positive for C. trachomatis. The frequency of C. trachomatis infection was 8.3% in the male partners of infertile couples at childbearing age when compared with 2.5% in the control group. It is most likely that infertility in the couples with chlamydial infection was due to the pathogen studied.
Assuntos
Infecções por Chlamydia/microbiologia , Chlamydia trachomatis/isolamento & purificação , Infertilidade/microbiologia , Feminino , Humanos , Infertilidade Masculina/etiologia , MasculinoRESUMO
We describe a nosocomial outbreak of extended-spectrum beta-lactamase (ESBL)-producing Serratia marcescens in a Bulgarian university hospital affecting nine patients on four wards. Phenotypic and genotypic (plasmid profile, randomly amplified polymorphic DNA analysis and amplified ribosomal DNA restriction analysis) analysis of the isolates indicated a single clone. The epidemic strain was resistant to oxyimino beta-lactams, aztreonam, aminoglycosides, tetracycline and chloramphenicol. It produced CTX-M-3 ESBL as demonstrated by isoelectric focusing, CTX-M PCR-RFLP and gene sequencing. The isolate was also found in the environment and from a nurse's hands, suggesting transmission by staff handling. The outbreak was controlled by patient isolation and intensified hand washing. This is the first report from Bulgaria describing a hospital outbreak caused by CTX-M-3 ESBL-producing S. marcescens.
Assuntos
Infecção Hospitalar/epidemiologia , Infecção Hospitalar/microbiologia , Surtos de Doenças , Infecções por Serratia/epidemiologia , Infecções por Serratia/microbiologia , Serratia marcescens/enzimologia , beta-Lactamases/isolamento & purificação , Adulto , Idoso , Antibacterianos/farmacologia , Técnicas de Tipagem Bacteriana , Bulgária/epidemiologia , Impressões Digitais de DNA/métodos , DNA Bacteriano/genética , Farmacorresistência Bacteriana , Microbiologia Ambiental , Genótipo , Mãos/microbiologia , Desinfecção das Mãos , Hospitais , Humanos , Transmissão de Doença Infecciosa do Profissional para o Paciente , Focalização Isoelétrica , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Enfermeiras e Enfermeiros , Isolamento de Pacientes , Plasmídeos , Infecções por Serratia/classificação , Serratia marcescens/isolamento & purificação , beta-Lactamases/químicaRESUMO
OBJECTIVES: Pneumococcal conjugate vaccines (PCVs) have reduced the incidence of pneumococcal disease, but non-vaccine serotypes are of concern, particularly if antimicrobial-resistant. This study retrospectively evaluated the serotype-specific clonality of paediatric multidrug-resistant (MDR) invasive and non-invasive Streptococcus pneumoniae isolates collected following PCV10 introduction (2011-2017) in Bulgaria. METHODS: Capsular types, drug resistance patterns and multilocus sequence typing (MLST) of the most common MDR S. pneumoniae serotypes sampled from children were determined. RESULTS: Overall, the rate of MDR pneumococci was 44.6% (107/240). The most common serotypes among MDR strains were 19F (25.2%), 19A (19.6%), 6C (13.1%), 6A and 23A (6.5% each) and 15A (4.7%), contributing 75.7% of all MDR strains. With the exception of serotype 19F, the remaining serotypes were non-PCV10 types. Among MDR pneumococci, the most frequently found sequence types were ST320 (30.4%; 19A and 19F), ST386 (12.7%; 6C and 6A) and ST8029 (5.1%; 23A). The majority of MDR STs (74.7%) belonged to PMEN clonal complexes, of which the most common were CC320 (Taiwan19F-14), CC315 (Poland6B-20) and CC180 (Netherlands3-31), accounting for 43.0%, 13.9% and 5.1%, respectively. In the post-vaccine period, a shift in the genetic structure of serotype 19A was found, with a significant increase of PMEN-14 (CC320) and a concurrent decrease of the major clone Denmark14-32 (CC230) observed prior to PCV10 introduction in Bulgaria. CONCLUSIONS: Clonality was found behind the wide distribution of MDR capsular types 19A, 6C, 23A and 3 following vaccine introduction, and a highly multiresistant and virulent clone Taiwan19F-14/ST320 has emerged as a common pathogen in children.
Assuntos
Infecções Pneumocócicas/prevenção & controle , Vacinas Pneumocócicas/administração & dosagem , Streptococcus pneumoniae/efeitos dos fármacos , Adolescente , Bulgária , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Tipagem de Sequências Multilocus , Infecções Pneumocócicas/imunologia , Infecções Pneumocócicas/microbiologia , Vacinas Pneumocócicas/genética , Vacinas Pneumocócicas/imunologia , Estudos Retrospectivos , Streptococcus pneumoniae/genética , Streptococcus pneumoniae/imunologia , Streptococcus pneumoniae/isolamento & purificação , VacinaçãoRESUMO
A total of 328 clinical isolates of Streptococcus pneumoniae were analyzed to determine the rate of macrolide and penicillin resistance as well as macrolide resistance phenotypes and genotypes. Erythromycin resistance was found in 81 pneumococcal isolates (24.7%) and 10.7% of isolates were clindamycin resistant. The prevalence of penicillin G-intermediate (minimum inhibitory concentrations, MICs, 0.125 to 1 microg/ml) and penicillin-resistant (MICs, >or=2 microg/ml) S. pneumoniae isolates was 25.6% and 13.7%, respectively. The rate of ceftriaxone-intermediate and ceftriaxone-resistant strains was 2.7% and 1.2%, respectively. Among erythromycin-resistant S. pneumoniae isolates, strains harboring mef(A) genes (n=42; 51.8%) were found to be predominant over strains with erm(B) genes (n=34; 42.0%). One (1.2%) isolate carried both erm(B) and mef(A), while 4 (4.9%) isolates carried L4 protein mutations. By using the erythromycin, clindamycin and rokitamycin triple-disk test, 42 strains were assigned to the M phenotype of macrolide resistance, 31 isolates were assigned to the partially inducible (iMcLS) phenotype, 4 were assigned to the constitutive (cMLS) phenotype. Four strains with L4 gene showed a rare phenotype with the triple-disk test. Serotyping of S. pneumoniae isolates suggested that serotype (or serogroup) 14, 6 and 19 were predominant (81.5%) among erythromycin-resistant strains. Among mef(A) positive isolates serotype 14 was predominant, among erm(B) positive isolates serogroups 6 and 19 were the most prevalent.
Assuntos
Antibacterianos/farmacologia , Farmacorresistência Bacteriana Múltipla/genética , Macrolídeos/farmacologia , Streptococcus pneumoniae/efeitos dos fármacos , Streptococcus pneumoniae/genética , Bulgária , Genes Bacterianos , Genótipo , Testes de Sensibilidade Microbiana , Fenótipo , Sorotipagem , Streptococcus pneumoniae/isolamento & purificaçãoRESUMO
A total of 132 ceftazidime-resistant clinical isolates of Pseudomonas aeruginosa were collected during 2001-2005 from 5 university hospitals in Sofia, Bulgaria to assess the current levels of antimicrobial susceptibility and to evaluate resistance mechanisms to beta-lactams. Antimicrobial susceptibilities were detected by a disk diffusion method and E-test. Polymerase chain reaction amplification and sequencing of bla(VEB-1 )and bla(PER-1 )were performed. The antibiotic resistance rates were: to piperacillin 90.2%, piperacillin/tazobactam 52.3%, ceftazidime 94.7%, cefepime 88.6%, cefpirome 98.5%, aztreonam 85.6%, imipenem 66.6%, meropenem 63.6%, amikacin 81.1%, gentamicin 84.8%, tobramycin 89.4%, netilmicin 57.6%, ciprofloxacin 83.4%. Structural genes for VEB-1 extended-spectrum beta -lactamases (ESBLs) were found in 75 (56.8%) of the isolates. PER-1 ESBLs were not detected. The VEB-1-producing strains were more resistant than VEB-1 non-producers to amikacin, gentamicin, tobramycin and ciprofloxacin ( P<0.001). VEB-1 appears to have a significant presence among ceftazidime-resistant P. aeruginosa isolates from Sofia.
Assuntos
Resistência às Cefalosporinas , Infecção Hospitalar/microbiologia , Infecções por Pseudomonas/microbiologia , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/enzimologia , beta-Lactamases/metabolismo , Antibacterianos/farmacologia , Bulgária/epidemiologia , Infecção Hospitalar/tratamento farmacológico , Infecção Hospitalar/epidemiologia , Proteínas de Escherichia coli , Humanos , Testes de Sensibilidade Microbiana , Infecções por Pseudomonas/tratamento farmacológico , Infecções por Pseudomonas/epidemiologia , beta-Lactamas/farmacologiaRESUMO
OBJECTIVE: To establish the incidence of cervicovaginal infections in patients with preterm and term deliveries. MATERIALS AND METHODS: 88 pregnancies with no previous detectable risk factors have been included in a retrospective study. 48 patients delivered at term and 40 had delivery between 24 and 37 weeks of gestation. A vaginal swab for bacterial infections and a cervical swab for detection of Chlamydia trachomatis infection, using PCR, have been performed in each case. The vaginal ecosystem has been appreciated according Nungent criteria. RESULTS AND CONCLUSIONS: C. trachomatis has been detected in 5% (2/40) of preterm deliveries and in 2.08% (1/48) of term deliveries. In preterm delivery group bacterial vaginosis has been isolated in 15% (6/40), Enterococcus spp. in12.5% (5/40) and Group B Streptococcus in 12.5% (5/40) of cases. In patients, delivered at term, bacterial vaginosis has been proved in 6.25% (3/48), Enterococcus spp in 8.33% (4/48), Group B Streptococcus in 4.17% (2/48) patients. Significant differences between both groups have been found in GBS and BV (p < 0.05). Although the difference C. trachomatis infection is not significantly higher in preterm delivery group, testing for C. tracomatis, using highly specific methods, along with vaginal swab are recommended as routine tests in pregnancy.
Assuntos
Complicações Infecciosas na Gravidez , Nascimento Prematuro/etiologia , Cervicite Uterina/complicações , Vaginite/complicações , Estudos de Casos e Controles , Feminino , Humanos , Gravidez , Complicações Infecciosas na Gravidez/microbiologia , Estudos Retrospectivos , Fatores de Risco , Cervicite Uterina/microbiologia , Vaginite/microbiologiaRESUMO
The biogenic iron oxide/hydroxide materials possess useful combination of physicochemical properties and are considered for application in various areas. Their production does not require special investments because these compounds are formed during cultivation of neurophilic iron bacteria. Bacteria from genus Leptothrix develop intensively in the Sphaerotilus-Leptothrix group of bacteria isolation medium and feeding media of Fedorov and Lieske. These media are different in their composition which determined the present study as an attempt to clear up the reasons that define the differences in the composition of the laboratory-obtained biomasses and the natural biomass finds. FTIRS, Mössbauer spectroscopy, and XRD were used in the research. Comparative analysis showed that the biomass and control samples contain iron compounds (α-FeOOH, γ-FeOOH, ß-FeOOH, γ-Fe2O3) in different ratios. The biomass samples were enriched in oxyhydroxides of higher dispersion. Organic residuals of bacterial origin, SO4, CO3, and PO4 groups were registered in the biogenic materials.
Assuntos
Biomassa , Meios de Cultura/química , Compostos de Ferro/química , Ferro/química , Leptothrix/crescimento & desenvolvimento , Minerais/químicaRESUMO
An oral polybacterial immunomodulator Urostim (U), composed of killed cells and their lysates from E. coli expressing type 1 and Pili, E. coli Rc mutant, P. mirabilis, K. pneumoniae and E. faecalis was created for immunoprophylaxis and immunotherapy of urinary tract infections (UTIs). In experimental animal models, the stimulating effect of U on lymphocyte functional activity, macrophage phagocytosis and antibody producing cells, was established. In this study the immuno-modulating effects of U on the proliferating capacity and ultrastructural morphologic changes of lymphocytes, cytokine production and specific systemic humoral and mucosal immune responses in patients with UTIs have been evaluated. Patients enrolled in the study, received orally 50 mg U daily for a period of three months. On days 0, 30 and 90 a quantitative analysis was performed on lymphoproliferative responses to polyclonal mitogens, IL-2 and the specific antigen U, the production of specific serum and saliva IgA, IgM and IgG antibodies to all components of U and the concentration of pro-inflammatory cytokines. There was significant improvement of non-specific and specific lymphoproliferative responses on days 30 and 90 after the onset of treatment with U, confirmed by electronmicroscopic studies. The highest concentrations of serum proinflammatory cytokines TNF-alpha, IL-1beta and IL-6 were registered at baseline followed by a decrease until the end of the observation period. This finding correlates with the gradual decrease of immune activation as measured by the spontaneous lymphocyte proliferation. Data from the production of specific antibacterial antibodies in serum and saliva show two types of reactions. The first type was registered in patients with low pre-treatment levels in whom the concentration of specific antibodies increased on days 30 and 90. The second type of reaction was observed in patients with high pre-treatment levels, which dropped on day 30 and were usually followed by an increase at the end of the study. These results provide evidence for the immuno-modulating effect of U. Our data show that the oral administration of the polybacterial immunomodulator Urostim stimulates adequate cellular and humoral systemic and mucosal immune responses in patients with chronic UTIs.
Assuntos
Adjuvantes Imunológicos/uso terapêutico , Vacinas Bacterianas/uso terapêutico , Infecções Urinárias/imunologia , Infecções Urinárias/terapia , Vacinas Combinadas/uso terapêutico , Adjuvantes Imunológicos/administração & dosagem , Administração Oral , Adolescente , Adulto , Idoso , Formação de Anticorpos , Vacinas Bacterianas/administração & dosagem , Técnicas de Cultura de Células , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Doença Crônica , Citocinas/biossíntese , Citocinas/sangue , Feminino , Humanos , Imunidade Celular , Imunidade nas Mucosas , Interleucina-2/farmacologia , Linfócitos/efeitos dos fármacos , Linfócitos/fisiologia , Linfócitos/ultraestrutura , Masculino , Pessoa de Meia-Idade , Mitógenos/farmacologia , Fatores de Tempo , Vacinas Combinadas/administração & dosagemAssuntos
Portador Sadio/fisiopatologia , Infecções por Coronavirus/fisiopatologia , Infecções por Coronavirus/transmissão , Vetores de Doenças , Fator C1 de Célula Hospedeira/isolamento & purificação , Pneumonia Viral/fisiopatologia , Pneumonia Viral/transmissão , Síndrome Respiratória Aguda Grave/fisiopatologia , Síndrome Respiratória Aguda Grave/transmissão , Animais , Betacoronavirus , COVID-19 , Chaperona BiP do Retículo Endoplasmático , Humanos , Pandemias , SARS-CoV-2RESUMO
Six hybridoma clones (1M, 4M, 9M, 11M, 18M and 31G), secreting monoclonal antibodies (mAbs) against lipid A were obtained after fusion between cells of mouse myeloma line X63-Ag8.653 and spleen cells from BALB/c mice immunized with acid treated Salmonella minnesota bacteria coated with additional free lipid A. The specificity and cross-binding activity of the mAbs were characterized in ELISA by using synthetic lipid A analogs as well as different lipid A and lipopolysaccharides (LPS) extracted from R- and S-form bacteria. It was found that the antibodies recognize epitopes in which phosphate groups, especially those at the C4' position of the glucosamine backbone of lipid A, were present. These epitopes were accessible also for the antibodies in purified intact LPS. By using a set of core glycolipids with increasing completion of the core region of the molecule and S-LPSs it was shown that the mAbs cross-reacted with a variety of R- and S-form LPS. The binding activity decreased with increasing length of the polysaccharide chain. The mAb did not prevent ultimate lethality of mice challenged with Klebsiella pneumoniae B and Salmonella typhimurium C5. However a delay of mortality rate of mice pretreated with antibodies 18M and 31G and infected with K. pneumoniae was seen.
Assuntos
Anticorpos Monoclonais/metabolismo , Lipídeo A/imunologia , Lipopolissacarídeos/imunologia , Animais , Sequência de Carboidratos , Reações Cruzadas , Ensaio de Imunoadsorção Enzimática , Feminino , Hibridomas/imunologia , Lipídeo A/análogos & derivados , Lipídeo A/química , Masculino , Camundongos , Camundongos Endogâmicos BALB C/imunologia , Camundongos Endogâmicos C57BL/imunologia , Dados de Sequência Molecular , Salmonella/imunologiaRESUMO
Seven hybridoma clones, secreting monoclonal antibodies (MAbs) against the genus-specific chlamydial lipopolysaccharide (LPS) antigen were obtained after immunization of BALB/c mice with formalin killed Chlamydia psittaci. The antigen-binding properties of the MAbs were characterized in different immunologic reactions with purified chlamydial elementary bodies and LPS antigens from S- and R-forms of Gram-negative bacteria. Four MAbs reacted with the heterologous LPS antigens of Salmonella R-mutants, Escherichia coli Re chemotype and Acinetobacter calcoaceticus. Two MAbs demonstrated in addition a significant reactivity with Porphyromonas gingivalis, E. coli O119 and Salmonella newington LPS in ELISA, dot-ELISA and passive hemolysis assay (for clone 204G9). One MAb cross-reacted only with Salmonella minnesota Re LPS in ELISA. In indirect immunofluorescent assay six MAbs produced bright green fluorescence with all tested chlamydial strains and five of them reacted with the Re and Rb2 chemotypes of S. minnesota. The results demonstrate a wide cross-reactivity of the produced MAbs with LPS antigens of various Gram-negative bacteria, posing the question for careful consideration and interpretation of serology results for Chlamydia spp.
Assuntos
Anticorpos Antibacterianos/imunologia , Infecções por Chlamydia/diagnóstico , Chlamydophila psittaci/imunologia , Escherichia coli/imunologia , Lipopolissacarídeos/imunologia , Porphyromonas gingivalis/imunologia , Salmonella/imunologia , Animais , Anticorpos Monoclonais/biossíntese , Anticorpos Monoclonais/imunologia , Infecções por Bacteroidaceae/diagnóstico , Infecções por Bacteroidaceae/imunologia , Infecções por Bacteroidaceae/microbiologia , Infecções por Chlamydia/imunologia , Infecções por Chlamydia/microbiologia , Ensaio de Atividade Hemolítica de Complemento/métodos , Reações Cruzadas , Ensaio de Imunoadsorção Enzimática/métodos , Infecções por Escherichia coli/diagnóstico , Infecções por Escherichia coli/imunologia , Infecções por Escherichia coli/microbiologia , Técnica Indireta de Fluorescência para Anticorpo , Bactérias Gram-Negativas/imunologia , Humanos , Immunoblotting/métodos , Camundongos , Camundongos Endogâmicos BALB C , Infecções por Salmonella/diagnóstico , Infecções por Salmonella/imunologia , Infecções por Salmonella/microbiologiaRESUMO
Macrolide resistance trends were examined among Helicobacter pylori strains from 154 patients between 1994 and 1998. Applicabilities of screening agar method (SAM) and modified disk diffusion method (MDDM) were evaluated. Overall primary resistance rates to erythromycin and clarithromycin were 14.8% and 8.7%, respectively. No association was found with age, sex, and diseases. Clarithromycin-resistance rate reached 12.5% in the last 2 years. Secondary resistance to erythromycin occurred more often (in 62.5%) than to clarithromycin (in 42.9%). Therapy with spiramycin or erythromycin in four cases induced no clarithromycin resistance. These data show a considerable prevalence of H. pylori resistance to macrolides, which exhibited a tendency to increase and was often associated with metronidazole resistance. By comparing the MDDM with SAM, an overall agreement was obtained in 81 (94.2%) of 86 results. MDDM and SAM are reliable techniques for testing H. pylori susceptibility to macrolides.
Assuntos
Antibacterianos/farmacologia , Claritromicina/farmacologia , Eritromicina/farmacologia , Helicobacter pylori/efeitos dos fármacos , Adolescente , Adulto , Bulgária , Criança , Pré-Escolar , Resistência Microbiana a Medicamentos , Feminino , Helicobacter pylori/crescimento & desenvolvimento , Helicobacter pylori/isolamento & purificação , Humanos , Masculino , Testes de Sensibilidade Microbiana , Fatores de TempoRESUMO
Hybridomas secreting immunoglobulin A (IgA) monoclonal antibodies (MAbs) against Salmonella enteritidis lipopolysaccharide (LPS) were generated after mucosal immunization of BALB/c mice with heat killed bacteria. Antigen binding properties and specificity of the produced MAbs were studied in ELISA and immunoblotting with purified LPS. Two IgA MAbs agglutinated all Salmonella OD1 strains and all S. enteritidis clinical isolates. MAb 178H11 recognized O:9 antigen of subserogroup OD1 LPS. MAb 177E6/A9 reacted also with OD3 LPS antigen and agglutinated OD3 strains. These data suggest the existence of different O:9 antigen subspecificities, one presented in subgroup OD1 and the other common for OD1 and OD3. Thus the produced IgA MAbs prove to be useful reagents, which could differentiate OD1 and OD3 from OD2 strains.
Assuntos
Anticorpos Antibacterianos/imunologia , Anticorpos Monoclonais/imunologia , Imunoglobulina A/imunologia , Salmonella enteritidis/imunologia , Animais , Anticorpos Antibacterianos/biossíntese , Anticorpos Monoclonais/biossíntese , Especificidade de Anticorpos , Antígenos de Bactérias/imunologia , Ensaio de Imunoadsorção Enzimática , Epitopos/imunologia , Immunoblotting , Lipopolissacarídeos , Camundongos , Camundongos Endogâmicos BALB C , Infecções por Salmonella/imunologia , Salmonella enteritidis/classificação , Salmonella enteritidis/patogenicidade , Sensibilidade e Especificidade , Sorotipagem/métodosRESUMO
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.