The role and place of serologic research in the system of epidemiologic surveillance of meningococcal infection

The results of serological surveys, carried out with a view to the detection of capsular meningococcal polysaccharides in the passive hemagglutination (PHA) test and covering 1,289 patients with systemic meningococcal infection, 221 patients with bacteriologically confirmed meningococcal nasopharyngitis, 2,820 persons in organized groups of children and adults with different epidemiological situation (including 650 carriers) and 4,050 residents of 8 cities with different morbidity levels, were analyzed. The patients, the carriers and the members of organized groups underwent multiple surveys. As shown in this study, the confirmation of the meningococcal etiology of the systemic forms of meningococcal infection (meningitis, meningococcemia) in all cases with the exception of children under 1 year of age, as well as the determination of the groups of meningococci, could be achieved in the PHA test twice as frequently, i.e. in 60-70% of cases, as with the use of the bacteriological method. The same regularities were observed in nasopharyngitis. The conclusion was made on the possibility of using the PHA test for more exact determination of the group of meningococci, prevailing at the initial stage of diseases, this determination playing an essential role in the epidemiological surveillance. The data obtained as the result of serological surveys in organized groups may lead to conclusions on the circulation of meningococci and their group composition, thus showing the necessity of profound epidemiological study with ensuring epidemic-control measures. The selective serological study of some hundreds of donor sera is not expedient in big cities, as it provides no data on the circulation of meningococci there.(ABSTRACT TRUNCATED AT 250 WORDS)     

Epidemiological significance of carriers of hyaluronidase-active meningococci

To find out the epidemiological role of the carriers of hyaluronidase-positive meningococcal strains, their spread in the foci of meningococcal infection, as well as outside these foci, was studied. For this purpose, altogether 5059 persons from 40 family foci of infection and 40 organized groups were examined. The number of carriers of hyaluronidase-active meningococci, detected among those of them who had contacts with sick persons, was 13 times greater than among the persons having no such contacts. In preschool institutions with unfavorable morbidity situation this number was 16 times greater than in those with favorable situation. In the family foci carriers of hyaluronidase-active meningococci constituted 80-88 %. The percentage of the carriers of hyaluronidase-active meningococci causing cases of the generalized form of meningococcal infection varied in different groups. According to the results obtained in this study, the threshold percentage of such carriers constituted 30 in preschool institutions, 36 in boarding schools and 50 in common school and in hostels for adults.     

Increasing the effectiveness of immunotherapy of rabies by using rifampicin in experimental studies]

The experiments showed that in a dose of 35-70 mg/kg rifampicin inhibited reproduction of the fixed rabies virus in the brain of infected animals. The drug had no inhibitory effect on synthesis of the virus-neutralizing antibodies after vaccination. Combination of rifampicin with antirabies gamma-globulin had a marked synergistic effect. The animal survival after the combination use amounted to 75-100 per cent and depended on the infective dose of the virus and the scheme of the drug administration. It was concluded that rifampicin might be used in complex therapy of rabies during the incubation period (along with gamma-globulin and the vaccine) for inhibiting virus reproduction at early infection stages.     

Characterization of the foci of meningococcal infection in closed groups of males

A total of 257 foci of meningococcal infection in groups of servicemen were selectively examined in 1982-2002. From these groups 353 patients with generalized forms of the disease underwent hospitalization. Most of the foci had a single infection, the proportion of foci with 10-40 patients was 82.6%. The meningococcal infection rate among humans in the foci was 25-37%, group A meningococci playing the leading role. In the structure of meningococcal infection generalized forms of infection constituted 16%, localized forms constituted 25% and inapparent forms (carriers)--59%. The formation of the morbidity structure was influenced by the type of the focus (with a single or multiple infection) and the character of morbidity for many years (during epidemic or at the period between epidemics). No absolute dependence of the level of meningococcal carrier state in the groups of servicemen on the appearance of the generalized forms of meningococcal infection was noted. Thus, both during epidemic and at the period between epidemics the population of meningococci, heterogeneous in its serological structure and differing in its clinical and epidemiological importance, constantly circulated with the leading role played by group A meningococci.     

Epidemiological characteristics of meningococcal infection in Cuba

The results of the epidemiological analysis of the morbidity rate in meningococcal infection for 1976-1984 are presented. The maximum rise of morbidity rate, equal to 14.4 per 100000 of population, was observed in 1983. Primarily, the rise of morbidity rate in 1979 was induced by meningococci of two serogroups: C (44.6%) and B (36.4%). The vaccinal prophylaxis of the population, carried out in 1979 with the use of polysaccharide vaccine A + C, did not affect morbidity caused by group B meningococci. The isolation rate of these organisms reached 98.7% from patients and 81.0% from carriers. The characteristic feature of the epidemic process of meningococcal infection in Cuba was a considerable increase in the number of patients under 1 year of age and the absence of seasonal fluctuations in morbidity rate.     

Suppression of the adhesive properties of mucosal microflora by ampicillin in peptic ulcer

Persistence of mucous bacteria in the mucosa of the stomach and duodenum was studied bacteriologically and electron microscopically in 132 patients with peptic ulcer. 235 microbial strains were isolated from the mucosa and its surface in 96 per cent of the patients. 80.8 per cent of the isolates belonged to different species of Staphylococcus and Streptococcus. The other 19.2 per cent of the isolates belonged to 9 rare bacterial species. 235 antibioticograms were analysed and it was shown that the predominating part of the isolates was mainly sensitive to ampicillin. Then follow ampiox, dicloxacillin and benzylpenicillin. Ampicillin was recommended for preoperative sanation of the stomach mucosa. 30 patients were given ampicillin orally in a dose of 0.25 g 4 times a day for 3 days before operations. It was found that the drug inhibited the adhesive properties of the mucous bacteria and even killed them.     

Sero-epidemiology of Neisseria meningitidis infections in parts of Eastern Nigeria.

The prevalence of antibodies to meningococci in the Nsukka area, East Central Nigeria, was studied by the indirect hemagglutination test employing tanned chicken erythrocytes. Of the 1332 apparently healthy persons, whose sera were tested, 982 samples (73.8%) were positive at 1:8 dilution of the sera. Fifty-six per cent of the population had antibodies to Group B, followed by 13% of Group C and 5% to Group A. Twenty-five sera from patients with meningococcal infections, from whom Group B meningococci were isolated, 100% were positive. Percentage reactivity in the 2-5-year age group was low (15%), whereas it increased rapidly from 78% (6-10-year group) to 95% (31 years and above).     

Serotypes among Neisseria meningitidis serogroups B and C strains isolated in Canada

Antisera made to prototype serogroup B strains of Neisseria meningitidis were used to serotype, by agar gel double diffusion, 262 meningococcal serogroups B and C strains isolated in Canada. The strains included 93 from patients and 169 from carriers. Serotype 2 was associated with 39 of 75 (52%) of group B strains and 14 of 18 (77.8%) of group C strains isolated from patients. The group B strains were mainly (87.2%) serotype 2b, while the majority (92.2%) of group C strains was serotype 2a. Other serotypes (including a new provisional serotype) represented 25.3 and 5.5% of groups B and C strains, respectively. The new serotype accounted for 13% of the group B strains. Approximately 23% of the strains isolated from patients were nontypable. The distribution of serotype 2, nontype 2 (other serotypes), and nontypable strains isolated from carriers was 2.1, 36.6, and 61.3%, respectively, for group B meningococci and 22.2, 29.6, and 48.25, respectively, for group C meningococci. Serotype 11 was the most prominent of the strains isolated from carriers. Approximately 7% of all the strains were multiple serotypes. Serotype 2 is an important virulence marker associated with meningococcal groups B and C disease in Canada, with serotypes 2a and 2b being markedly associated with groups C and B meningococcal disease, respectively.     

Phage variation: understanding the behaviour of an accidental pathogen

Understanding why carriers of meningococci occasionally develop invasive disease is a major challenge. Individual strains of meningococci are extremely variable and undergo dynamic changes in DNA content and organization. This heterogeneity of meningococcal populations might enhance the fitness of this human-restricted bacterium. The recent discovery of a meningococcal bacteriophage and its associations to disease is an intriguing example of this variability and could contribute to a better understanding of microbial commensal and virulence behaviour.     

Nucleotide sequence analysis of the sialyltransferase genes of meningococcal serogroups B,C, Y and W135

A rapid method for serogrouping meningococci is essential for the characterization of phenotypically non-groupable meningococcal isolates and clinical samples, particularly for public health management purposes. The Scottish Meningococcus and Pneumococcus Reference Laboratory (SMPRL) provides serogrouping results of meningococcal isolates and clinical samples using a PCR assay which detects restriction fragment length polymorphisms in meningococcal serogroups B, C, Y and W135. Although this PCR system was invaluable when first introduced, it has several drawbacks and lacks the required sensitivity for detecting DNA in clinical samples. Due to the recent introduction of the meningococcal group C conjugate vaccine and an impending group B vaccine, a more robust and informative method for serogroup determination is required. A protocol was devised allowing PCR amplification of the siaD gene of serogroup B, C, Y and W135 meningococci. This system was multiplexed and allowed serogroup differentiation between serogroups B and C and also between B/C and Y/W135 by product size analysis. A nested stage was incorporated into the system for enhanced detection of meningococci in clinical samples, and finally a sequencing protocol was designed allowing detection of any nucleotide changes within the siaD gene. This system allows rapid serogrouping results for use within an agarose gel system as well as more informative results when used for sequencing within the siaD gene.     

Mannose binding lectin enhances IL-1beta and IL-10 induction by non-lipopolysaccharide (LPS) components of Neisseria meningitidis

Mannose binding lectin (MBL) is a key molecule in the lectin pathway of complement activation, and likely of importance in our innate defence against meningococcal infection. We evaluated the role of MBL in cytokine induction by LPS or non-LPS components of Neisseria meningitidis, using a meningococcal mutant deficient for LPS. Binding experiments showed that MBL exhibited low, but significant binding to encapsulated LPS+ meningococci (H44/76) and LPS-deficient (LPS-) meningococci (H44/76lpxA). Experiments with human mononuclear cells (PBMCs) showed that MBL significantly augmented IL-1beta production after stimulation with LPS+ and LPS- meningococci, in a dose-dependent fashion. In addition, IL-10 production was enhanced after stimulation with LPS- meningococci. In contrast, TNFalpha, IL-6 and IFNgamma productions were unaffected. No effect of MBL was observed on cytokine induction by meningococcal LPS. MBL enhanced cytokine production at concentrations >10(7) meningococci. It is concluded that MBL interacts with non-LPS components of N. meningitidis and in this way modulates the cytokine response.     

The epidemiological characteristics of meningococcal infection in the USSR

The epidemiological analysis of morbidity in meningococcal infection in the USSR in the period of 1969-1987 showed that the second rise of the morbidity level occurred in 1984 and was followed by its decrease in most of the regions of the USSR. This study also revealed that the characteristic feature of the second rise of morbidity in meningococcal infection was a considerable involvement of young children (aged up to 3 years), as well as the increased etiological role of group B meningococci in cases of meningococcal infection and the circulation of these microorganisms among the population. Besides, the preservation of the etiological importance of group A meningococci in many regions of the USSR, especially among adults, was noted. In this connection, the use of Soviet group A meningococcal polysaccharide vaccine on epidemiological indications was considered to be epidemiologically substantiated.     

Assessment of the sensitivity of Neisseria meningitidis strains to benzylpenicillin using the disc diffusion method]

The sensitivity of 235 N. meningitidis strains to 5 antibiotics was estimated by the diameter of growth inhibition zones according to the criteria recommended by a Laboratory (Marseilles, France) collaborating with the WHO. All the strains proved to be sensitive to benzylpenicillin when disks containing 10 and 2 IU of the antibiotic were used. The strains were also shown to be sensitive to chloramphenicol and tetracycline. 95.7 and 7.7 per cent of the strains were sensitive to rifampicin and oleandomycin, respectively. When the strain sensitivity was assayed with the disks containing 10 and 2 IU of benzylpenicillin by the more severe criteria recommended by J. Saez-Nieto et al., significant changes were detected: meningococci with relative resistance to benzylpenicillin were detected in various regions of this country and the number of such strains was found to have a tendency to slightly increase.     

Prevention of secondary cases of meningococcal disease in household contacts by vaccination.

Household contacts of patients with group A meningococcal infection were vaccinated with either meningococcal vaccine or tetanus toxoid. Five of the 523 subjects who received tetanus toxoid developed meningococcal meningitis and another four probably had meningococcal disease. Only one possible case of meningococcal infection occurred among 520 contacts vaccinated with meningococcal vaccine. Vaccination had no effect on nasopharyngeal carriage of meningococci. Vaccination of household contacts of patients with group A meningococcal infections is an effective way of using limited supplies of meningococcal vaccine, though its value would be limited in an epidemic. Secondary cases of meningococcal infection often occur within a few days of the index case, and, although vaccine alone seemed to provide adequate prophylaxis in these Nigerian subjects, additional chemoprophylaxis may be needed to cover this critical period.     

The immunological status indices of monkeys inoculated with a meningococcal B vaccine

The method for the determination of bacterial antibodies to group B meningococci was worked out. The method was used for the determination of antibodies to group B meningococcal vaccine produced in the USSR. The dynamic study of antibodies to protein, polysaccharide and lipopolysaccharide antigens of group B meningococci was made by the method of the enzyme immunoassay (EIA), and the safety of the vaccine was studied by the determination of autoantibodies active against brain tissue antigens. The data thus obtained were indicative of the immunological activity of group B protein-polysaccharide vaccines, manifested by the capacity for stimulating bactericidal antibodies whose level increased 8- to 10-fold after the immunization of monkeys in 2 and 3 injections. Similarity in the dynamics of the formation of bacteriolysins and antibodies to protein antigen, as determined in EIA, was noted. The vaccine was found to stimulate no cytotoxic anticerebral antibodies in the glia migration test, which was indicative of the safety of group B meningococcal vaccine.     

Isolation and low-temperature preservation of liposomes containing rifampicin

The optimal conditions for preparations of rifampicin-containing liposomes were determined with the methods of mechanical shaking, gas dispersion and and reversible phases. It was found that the percentage of rifampicin incorporation into liposomes depended on the molar ratio of the antibiotic to the lipid (the optimal ratio was 1 : 10), the size and structure of liposomes, the amount of cholesterol added and the lipid membrane charge. Incorporation of rifampicin amounted to 16.1 +/- 2.4, 39.2 +/- 3.2 and 60.5 +/- 2.9 per cent with respect to neutral lecithin multilamellar liposes, liposomes prepared with the gas dispersion method and liposomes prepared with the method of reversible phases, respectively. Cholesterol in a molar ratio to lecithin equal to 2 : 5 or higher and dicetyl phosphate imparting the negative charge to the membrane had an inhibitory effect on the drug uptake by liposomes, while stearyl amine having the positive charge had a stimulating effect. The effect of the cryoprotectors glucose, polyvinylpyrrolidone, poly-ethylene glycole-400 and glycerol on low-temperature preservation and storage of rifampicin-containing liposomes was studied. It was shown that 10--15 per cent solutions of sucrose and glucose had the highest cryoprotective effect, when the two-stage method of freezing was used. It provided almost 84 per cent preservation of liposomal rifampicin. Electron microscopy showed that after defrosting liposomes no significant changes in the size and structure of lipid membranes were observed.     

Novel blocking human IgG directed against the pentapeptide repeat motifs of Neisseria meningitidis Lip/H.8 and Laz lipoproteins

Ab-initiated, complement-dependent killing contributes to host defenses against invasive meningococcal disease. Sera from nonimmunized individuals vary widely in their bactericidal activity against group B meningococci. We show that IgG isolated from select individuals can block killing of group B meningococci by human sera that are otherwise bactericidal. This IgG also reduced the bactericidal efficacy of Abs directed against the group B meningococcal protein vaccine candidates factor H-binding protein currently undergoing clinical trials and Neisserial surface protein A. Immunoblots revealed that the blocking IgG was directed against a meningococcal Ag called H.8. Killing of meningococci in reactions containing bactericidal mAbs and human blocking Abs was restored when binding of blocking Ab to meningococci was inhibited using either synthetic peptides corresponding to H.8 or a nonblocking mAb against H.8. Furthermore, genetic deletion of H.8 from target organisms abrogated blocking. The Fc region of the blocking IgG was required for blocking because F(ab')(2) fragments were ineffective. Blocking required IgG glycosylation because deglycosylation with peptide:N-glycanase eliminated blocking. C4b deposition mediated by an anti-factor H-binding protein mAb was reduced by intact blocking IgG, but not by peptide:N-glycanase-treated blocking IgG, suggesting that blocking resulted from inhibition of classical pathway of complement. In conclusion, we have identified H.8 as a meningococcal target for novel blocking Abs in human serum. Such blocking Abs may reduce the efficacy of select antigroup B meningococcal protein vaccines. We also propose that outer membrane vesicle-containing meningococcal vaccines may be more efficacious if purged of subversive immunogens such as H.8.     

The evolution of meningococcal infection in Mongolia]

On the basis of the generalization and analysis of the results of bacteriological and immunological investigations the epidemic process of meningococcal infection (MI) in Mongolia was found to undergo definite changes during the last 20 years. Group A meningococci prevailing in the etiology of MI were replaced by strains belonging to group B affecting mainly young children (aged up to 3 years). MI morbidity rate caused by group B meningococci was found to be higher in Mongolia than in other countries of the world. These data substantiate the necessity of using more effective remedies for the control of this infection and, in particular, specific immunization with vaccines against group B meningococci; profound study of the properties of the circulating meningococcal strains is to be carried out.     

Antibiotic sensitivity of Vibrio cholerae 01 isolated from environmental objects]

The sensitivity of 252 Vibrio cholerae-O1 strains isolated from environmental objects to antibiotics of various groups was assayed by the method of serial dilutions on solid media. The biological characteristics of the isolates are presented. The Vibrio cholerae isolates with serological variation were the most frequent (36.6 per cent), so are the cultures detected by their sensitivity to the specific phages (87.5 per cent). It was found that changes in some biological properties of the strains did not coincide with the changes in the antibiotic sensitivity. The isolates were highly sensitive to tetracycline, chloramphenicol, gentamicin, erythromycin and rifampicin and less sensitive to novobiocin and the other aminoglycosides. The sensitivity to the beta-lactams was the lowest. The resistance determinants were detected in single strains (6.3 per cent), the kanamycin and novobiocin resistance determinants being detected in 15 out of the 16 strains tested. The study showed that the cultures of Vibrio cholerae-O1 isolated from the environmental objects generally preserved their sensitivity to the diverse group antibiotics.     

Cellular ultrastructure of the meningococcus when incubated in a continuous culture of human FL-line amnion

Some details of the ultrastructure of several meningococcal strains having had contacts with cells in continuous human amnion cell culture FL for 6 hours to 2 days have been defined with greater precision by means of electron microscopy. The study has shown that the contact of meningococci with the tissue culture is accompanied by the appearance of meningococcal forms with the defective cell wall, similar to L-forms: spheroplast, protoplast, gigantic cells and microcells, as well as budding variants. The meningococcal variants with the defective cell wall, appearing in the cell culture, and the forms occurring (in different proportions) in "ripe" meningococcal populations developing in the culture media for a long time and isolated from a human body have been found to have no essential differences in their fine structure. These data indicate that any external influences (meningococci are highly sensitive to such influences) produce sufficiently rapid changes, similar to L-transformation, in the fine structure of these microorganisms.