Phenotypic and genetic characterization of antimicrobial profiles of Helicobacter pylori strains in Cuba.

The study evaluated the antibiotic resistance patterns of Helicobacter pylori strains against metronidazole and clarithromycin in a hospital in Havana, Cuba. Eighty-five percent, 22.5%, and 10% of 40 H. pylori strains investigated were resistant to metronidazole, ciprofloxacin, and clarithromycin respectively but all were susceptible to amoxicillin and tetracycline. RdxA truncation was found only in metronidazole-resistant strains. In such strains, reported are eight and two novel mutations in the rdxA and frxA genes respectively. Two-point mutations in the 23S rRNA genes of clarithromycin-resistant strains were detected. A high prevalence of metronidazole resistance was found in Cuban H. pylori strains. Mutations in the rdxA gene may contribute more significantly than frxA gene to the high level of resistance to metronidazole. This study supports the need to continue monitoring the antibiotic susceptibility in H. pylori in Cuba to guide the treatment of such infection.

Safety and preliminary immunogenicity of MenC/P64k, a meningococcal serogroup C conjugate vaccine with a new recombinant carrier.

This study reports the preliminary assessment of the safety and immunogenicity of the first serogroup C conjugate vaccine candidate that includes meningococcal P64k recombinant protein as the carrier (MenC/P64k). Twenty volunteers were recruited for a double-blind, randomized, controlled phase I clinical trial, receiving a single dose of MenC/P64k (study group) and a single dose of the commercial polysaccharide vaccine AC (control group). Only mild reactions were observed. No statistical differences were detected between the antipolysaccharide C IgG responses of both groups as well as between bactericidal serum titre (P > 0.05). The MenC/P64k vaccine was found to have a good safety profile, to be well tolerated and immunogenic.

Evaluation of the BIOCEN GC agar medium base in antimicrobial susceptibility testing of Neisseria gonorrhoeae.

BACKGROUND: Since 1995, the Cuban Reference Laboratory for Neisseria has been monitoring the antibiotic susceptibility of gonococci, following the methodology of the National Committee for Clinical Laboratory Standards, which uses GC agar medium base supplemented with 1% Vitox. We evaluated three lots of GC agar medium base produced by BIOCEN, Cuba, in antibiotic susceptibility testing of reference and wild strains of gonococci. METHODS: The susceptibilities to five antibiotics were evaluated five times on three lots of GC agar medium base from BIOCEN. Four and one gonococcal reference strains were tested by MIC dilution and disc diffusion methods, respectively. Later, the antimicrobial susceptibilities of ten wild Neisseria gonorrhoeae strains were tested in triplicate. As internal control, a GC agar medium from Difco was used. RESULTS: All antibiotic MICs obtained on four lots of GC agar medium from different manufacturers fell within the proposed quality control limits for reference strains analyzed. The disc diffusion data for the reference strain of N. gonorrhoeae ATCC 49226 to five antibiotics provided essentially identical results in all lots of GC agar medium base. For wild strains of gonococci, identical modal MIC values and zone size diameters within a 3-mm range were observed in all the antibiotics tested. CONCLUSIONS: Excellent agreement in susceptibility testing methods among different lots of GC agar medium base from BIOCEN and Difco was obtained for all reference and wild gonococcal strains and antibiotics tested. We proposed that GC medium from BIOCEN can be used in antimicrobial susceptibility testing of N. gonorrhoeae by MIC dilution and disc diffusion tests.

Factibilidad del uso del medio de cultivo base de Agar GC-Biocen en el Laboratorio de Microbiología / Feasability of the Agar GC-Biocen base culture medium in the microbiology laboratory

En el laboratorio nacional de referencia de Neisserias del Instituto de Medicina Tropical"Pedro Kouri" de Cuba, se realiza desde el año 1995 un monitoreo sistemático de la susceptibilidad antimicrobiana de las cepas de N.Gonorrhoeae aisladas en diferentes provincias,donde se emplea como medio de cultivo la base de agar gonococcus, en todos los laboratorios de microbiologia de Cuba(AU)

Evaluation of four methods for detecting the beta-lactamase activity in Neisseria gonorrhoeae isolated in Cuba

Four methods (chromogenic, acidimetric, inhibition, and iodometric) for demonstration of the beta-lactamase production by 70 isolates of Neisseria gonorrhoeae, were evaluated in Cuba. There was 100 percent correlation between all beta-lactamase methods and the standardized penicillin dilution susceptibility test for penicillinase-non-producing N. gonorrhoeae. For penicillinase-producing N. gonorrhoeae strains, there was a perfect correlation between the chromogenic method and penicillin susceptibility testing, but one and two strains failed to give a positive result for beta-lactamase with the inhibition/acidimetric and the iodometric methods, respectively. There was a high concordance between the chromogenic method, considered as gold standard and the rest of penicillinase tests evaluated: Kappa Index (KI) = 0.98 for inhibition/acidimetric methods and KI = 0.97 for the iodometric method. The four methods evaluated were accurate, reproducible, easily readable, economical, and ease to use for screening primary isolates of N. gonorrhoeae in Cuba. We recommended the use of the inhibition method, when testing the penicillinase activity in gonococcal isolates in provincial and municipal reference laboratories.

Evaluación de diferentes métodos de utilización de azúcares en Neisseria gonorrhoeae / Evaluation of different methods of utilization of sugars in Neisseria gonorrhoeae

Se evaluaron 5 métodos de utilización de azúcares en 25 cepas identificadas previamente como Neisseria gonorrhoeae: agar CTA, agar CTA modificado, agar gelatina almidón, agar Mueller Hinton más azul de bromotimol y método rápido. Por los métodos de CTA y rápido fueron identificadas 100 por ciento de las cepas de N. gonorrhoeae, mientras que por los métodos de CTA modificado y agar gelatina almidón fue 96 por ciento. Ninguna cepa de gonococo fue identificada por el método de agar Mueller Hinton más azul de bromotimol. El medio de agar cistina tripticasa (CTA), constituye el método de elección para confirmar los aislamientos de Neisseria gonorrhoeae; es elaborado fundamentalmente por compañías norteamericanas, por lo que su adquisición en Cuba resulta difícil. Los métodos de agar gelatina almidón y rápido constituyen alternativas útiles del medio CTA, por lo que su uso se propone en este trabajo

Evaluación de diferentes métodos de utilización de azúcares en Neisseria gonorrhoeae / Evaluation of different methods of utilization of sugars in Neisseria gonorrhoeae

Se evaluaron 5 métodos de utilización de azúcares en 25 cepas identificadas previamente como Neisseria gonorrhoeae: agar CTA, agar CTA modificado, agar gelatina almidón, agar Mueller Hinton más azul de bromotimol y método rápido. Por los métodos de CTA y rápido fueron identificadas 100 por ciento de las cepas de N. gonorrhoeae, mientras que por los métodos de CTA modificado y agar gelatina almidón fue 96 por ciento. Ninguna cepa de gonococo fue identificada por el método de agar Mueller Hinton más azul de bromotimol. El medio de agar cistina tripticasa (CTA), constituye el método de elección para confirmar los aislamientos de Neisseria gonorrhoeae; es elaborado fundamentalmente por compañías norteamericanas, por lo que su adquisición en Cuba resulta difícil. Los métodos de agar gelatina almidón y rápido constituyen alternativas útiles del medio CTA, por lo que su uso se propone en este trabajo(AU)

Antimicrobial susceptibility of Neisseria gonorrhoeae in Cuba (1995-1999): implications for treatment of gonorrhea.

BACKGROUND: Antibiotic-resistant strains of Neisseria gonorrhoeae, especially those resistant to penicillin and tetracycline, have spread with remarkable rapidity in many Caribbean countries. GOAL: The goal of the study was to survey the antibiotic susceptibilities of N gonorrhoeae strains isolated from 1995 to 1999 in Cuba and to discuss the impact of antimicrobial resistance on the management of gonorrhea in the country. STUDY DESIGN: Susceptibility of the strains to penicillin, tetracycline, cefuroxime, ceftriaxone, ciprofloxacin, spectinomycin, and azithromycin were determined by an agar dilution method. RESULTS: Penicillin and tetracycline resistance was noted in 60.8% and 54.2%, respectively, of the N gonorrhoeae strains tested. A total of 63.35 (76/120) of the N gonorrhoeae strains exhibited plasmid-mediated resistance to penicillin, tetracycline, or both. Strains with chromosomally mediated resistance to these antibiotics accounted for 10% (12/120) of the strains. The strains were susceptible to ceftriaxone, cefuroxime, spectinomycin, and ciprofloxacin. One strain's ciprofloxacin MIC was 0.125 mircog/ml. Of the 52 strains tested, 23.1% displayed intermediate resistance to azithromycin. CONCLUSIONS: N gonorrhoeae strains exhibited a high frequency of resistance and multiresistance to penicillin and tetracycline. Therefore, these antibiotics should no longer be used to treat gonococcal infections in Cuba and should be substituted with effective drugs such as third-generation cephalosporins, spectinomycin, and fluoroquinolones. The detection of intermediate resistance to azithromycin and ciprofloxacin underlines the importance of periodic surveillance for susceptibility of N gonorrhoeae strains to antimicrobials agents used as primary therapy for gonorrhea.

[Evaluation of different methods of utilization of sugars in Neisseria gonorrhoeae]. / Evaluación de diferentes métodos de utilización de azúcares en Neisseria gonorrhoeae.

5 methods of utilization of sugars were evaluated in 25 strains previously identified as N. gonorrhoeae: CTA agar, modified CTA agar, gelatin starch agar, Mueller Hinton agar plus bromotimol blue and rapid method. 100% of the strains of N. gonorrhoeae were identified by the CTA and rapid methods, whereas 96% were identified by the modified CTA and gelatin starch agar methods. No strain of gonoccocus was identified by the Mueller Hinton agar method plus bromotimol blue. The cystine tripticase agar medium (CTA) is the elective method to confirm the isolates of Neisseria gonorrhoeae. As it is mainly made by U.S. companies, it is difficult for Cuba to acquire it. The gelatin-starch agar method and the rapid method are useful alternative of the CTA medium, so their use is proposed in this paper.

Evaluation of four methods for detecting the beta-lactamase activity in Neisseria gonorrhoeae isolated in Cuba.

Four methods (chromogenic, acidimetric, inhibition, and iodometric) for demonstration of the beta-lactamase production by 70 isolates of Neisseria gonorrhoeae, were evaluated in Cuba. There was 100% correlation between all beta-lactamase methods and the standardized penicillin dilution susceptibility test for penicillinase-non-producing N. gonorrhoeae. For penicillinase-producing N. gonorrhoeae strains, there was a perfect correlation between the chromogenic method and penicillin susceptibility testing, but one and two strains failed to give a positive result for beta-lactamase with the inhibition/acidimetric and the iodometric methods, respectively. There was a high concordance between the chromogenic method, considered as gold standard and the rest of penicillinase tests evaluated: Kappa Index (KI) = 0.98 for inhibition/acidimetric methods and KI = 0.97 for the iodometric method. The four methods evaluated were accurate, reproducible, easily readable, economical, and ease to use for screening primary isolates of N. gonorrhoeae in Cuba. We recommended the use of the inhibition method, when testing the penicillinase activity in gonococcal isolates in provincial and municipal reference laboratories.

Typing and susceptibility to penicillin of Neisseria meningitidis isolated from patients in Cuba (1993-1999)

The susceptibility to penicillin of 111 Neisseria meningitidis strains was assessed by the agar-dilution procedure and serosubtypes were determined by a whole-cell enzyme-linked immunoassay using monoclonal antibodies reagents. Thirty-five isolates showed reduced sensitivity to penicillin (MIC > or = 0.1 mg/l and <= 1 mg/l) and no resistant strains were detected. The most common phenotype was B:4:P1.15 (77.5 percent) and a rising trend of non-typeable and non-subtypeable strains was detected. The increase in levels of minimal inhibitory concentrations of meningococci to penicillin gives cause for concern and the increase in non-typeable and non-subtypeable isolation demand the use of molecular biology techniques for their typing

Characterization of Neisseria gonorrhoeae strains isolated from patients with conjunctivitis

The conjunctivitis produced by Neisseria gonorrhoeae is the less frequently reported clinical form of gonococcal infection. We aim to phenotypically characterize N. gonorrhoeae isolated from conjunctivae sites. A total of six cases of this disease were notified in the Camagüey province, Cuba. All the strains isolated were penicillin-producing, showed the serogroup WI and exhibited the same antimicrobial susceptibility pattern and plasmid profile (2.6-3.2-24.5). The results contribute to the characterization of N. gonorrhoeae strains circulating in our environment