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Identification of four fimbria-encoding genomic islands that are highly specific for verocytotoxin-producing Escherichia coli serotype O157 strains.

Verocytotoxin-producing Escherichia coli causes zoonotic food- or waterborne infection that may be associated with massive outbreaks and with the serious complication of hemolytic uremic syndrome (HUS). Serotypes O157:H7 and O157:NM are more commonly associated with HUS and outbreaks than other serotypes, such as O26:H11. To determine whether a genetic basis exists for why serotype O157:H7/NM causes HUS and outbreaks more often than other serotypes, such as O26:H11, we conducted suppression subtractive hybridization (SSH) between the genomes of the sequenced O157:H7 strain EDL933 and CL1, a clinical serotype O26:H11 isolate. Genes from four EDL933 fimbria-encoding genomic O islands (OIs) (OI-1, -47, -141, and -154) were identified in the SSH library. OI-47 encodes several additional putative virulence factors, including secreted and signaling proteins, a hemolysin locus, a lipoprotein, an ABC transport system, and a lipid biosynthesis locus. The distribution of the OIs was investigated by PCR and Southern hybridization (when PCR was negative) with 69 VTEC strains belonging to 39 different serotypes corresponding to 5 seropathotypes that differ in their disease and epidemic potential. The four OIs described here were distributed almost exclusively in serotypes O157:H7 and O157:NM, which indicates that they may be associated with the ability of these strains to colonize human and/or animal intestinal tracts and to cause epidemic and serious disease more frequently than other serotypes. The occurrence of the four OIs in enteropathogenic E. coli O55:H7 strains is consistent with their vertical inheritance by VTEC O157:H7/NM from this clonally related ancestor.

Escherichia coli O157↗

Characterization of a common high-affinity receptor for reovirus serotypes 1 and 3 on endothelial cells.

During viremia, viruses may be cleared from the bloodstream and taken up by specific organs. The uptake of virus from the bloodstream is dependent on the association of viral particles with endothelial cells that line the luminal surfaces of large and small blood vessels. To understand the nature of this interaction, we have studied the binding of reovirus serotypes 1 and 3 to these cells in vitro. Both serotypes of reovirus productively infected endothelial cells. By using [35S]methionine-biolabeled reovirus as a tracer ligand, we found that both viruses rapidly bind to endothelial cells and that equilibrium is reached after 4 h. The binding of the radiolabeled viruses was saturable and mediated by a homogeneous population of cellular receptors with very high affinity (Kd = 0.5 nM) for the virus ligands. Both serotypes bind to the same receptor, since the attachment of each radiolabeled serotype is inhibited by both the homologous and heterologous unlabeled virus. Exposure of labeled virus to monoclonal antibodies directed against the viral hemagglutinin (sigma 1 protein) inhibited binding, demonstrating that the attachment of reovirus to endothelial cells is mediated by the hemagglutinin for both serotypes. By using a novel ligand-blotting assay, the binding of both viruses to a 54,000-dalton protein could be demonstrated. The binding of each radiolabeled serotype to this protein was inhibited by the homologous and heterologous unlabeled serotype. By using cell fractionation after homogenization, we demonstrated that this 54-kilodalton protein is a membrane protein, in agreement with its proposed role as a cell surface receptor for reovirus serotypes 1 and 3.

Animals↗

Effect of passive immunization with serotype-specific monoclonal antibodies on Actinobacillus pleuropneumoniae infection of mice.

Specific monoclonal antibodies (MoAbs) to Actinobacillus pleuropneumoniae (APP) serotypes 1 and 2 which recognized serotype-specific antigens were produced. It was revealed that the two serotype-specific MoAbs H1-18 and H22-7 recognized O polysaccharides of the lipopolysaccharide (LPS) from APP serotypes 1 and 2, respectively, in the results of antigen analysis by means of SDS-PAGE and Western blotting. Furthermore, ddY mice immunized passively with the above type-specific MoAbs were protected against challenge infection by the homologous serotype of APP at 24 or 48 hr later. However, H1-9 and H3-2 MoAbs recognizing serotype-common protein antigens of APP did not show signs of any protective effect. These results showed that LPS from APP bacterial cells was one of the structural substances in the serotype-specific antigens, and an important component as one of the antigens protecting against the homologous serotype strain.

Actinobacillus Infections↗

Serotype distribution & antimicrobial resistance in Streptococcus pneumoniae causing invasive & other infections in south India.

BACKGROUND & OBJECTIVES: Streptococcus pneumoniae continues to be a major cause of morbidity and mortality in developing countries. The emerging resistance to some common antibiotics compounds the problem. There arises a need to monitor the resistance pattern and map serotype distribution in different geographic locations. The present study was undertaken to determine the serotype prevalence and antibiotic susceptibility of clinically significant S. pneumoniae isolated from a tertiary care hospital in south India. METHODS: A total of 150 clinical isolates from invasive and other clinically significant pneumococcal infections were serotyped and screened for susceptibility to commonly used antibiotics by standard and modified laboratory procedures. RESULTS: Majority (59.3%) of the isolates belonged to one or other of the serotypes 1, 6, 19, 5, 23 and 7. Serotype 1 was the commonest isolate from patients of meningitis and empyema followed by pneumonia. Nineteen isolates (12.6%) were nonvaccine type. Eleven (7.3%) isolates were relatively resistant to penicillin (minimum inhibitory concentration was between 0.1 and 1 microgram/ml) and 64 were resistant to one or more antibiotics. Resistance was distributed equally among the predominant serotypes. INTERPRETATION & CONCLUSION: The common serotypes responsible for significant infections were similar to those reported in some other studies from India, with minor variations. Resistance to cotrimoxazole and tetracycline was predominant followed by chloramphenicol. Low level resistance to penicillin was observed but no isolate had absolute resistance. This calls for monitoring of resistance and mapping of serotype distribution from various parts of India.

Adolescent↗

[Serotypes of hepatitis C virus in mixed cryoglobulinaemia].

BACKGROUND: Several authors have studied the genotypic distribution of hepatitis C virus (HCV) in mixed cryoglobulinaemia (MC), which represents the most typical extra-hepatic manifestation of the HCV infection. On the other hand, at present no data are available on the HCV serotypic characterization of MC patients. METHODS: Thus, 28 serum specimens from HCV positive patients affected by MC (7 males and 21 females; mean age of 64 yrs, range 35-80) have been evaluated by a serotyping EIA method (Murex HCV Serotyping 1-6 Assay, Murex Diagnostics, Pomezia, Italy). RESULTS: The HCV serotype 1 was found in 13 patients (46.4%), serotype 2 in 5 patients (17.9%) and mixed serotypes in 3 patients (10.7%), showing, respectively, serotypes 1+2, 1+4, and 1+4+5. Moreover, the HCV serotype was not detectable in 7 patients (25.0%). CONCLUSIONS: These results, obtained by using a simple, serologic technique, agree with some literature data concerning MC patients from the same geographic region as ours (Venetian area). In addition, our findings appears to reflect merely the genotypic distribution of the HCV infection in the North Italy.

English Abstract↗

Preparation of a neutralizing monoclonal antibody specific to serotype 8 rotavirus strains with a super-short RNA pattern.

Extensive cross-neutralization tests have identified seven serotypes of rotavirus. Recently, it has been reported that several human strains (69M, B37 and B38) with a super-short RNA pattern carry new serotype specificity (serotype 8). By using the strain 69M as an immunizing antigen, we prepared a neutralizing monoclonal antibody (designated 69M-2D). In neutralization tests using 18 human and two animal rotavirus strains with different serotype specificities, the 69M-2D antibody was found to neutralize only serotype 8 strains. Also, in an enzyme-linked immunosorbent assay (ELISA), the antibody reacted exclusively with the serotype 8 strains. Thus, the reactivity pattern of the 69M-2D antibody was serotype 8-specific. This antibody should be useful as another reagent for serotyping rotaviruses by ELISA.

Animals↗

Cross protection in mice and swine immunized with live erysipelas vaccine to challenge exposure with strains of Erysipelothrix rhusiopathiae of various serotypes.

Mice and swine immunized subcutaneously with live vaccine prepared from acriflavine-fast attenuated Erysipelothrix rhusiopathiae, strain Koganei (serotype 2), were challenge exposed to virulent strains of E rhusiopathiae of various serotypes. Vaccinated mice did not die after challenge exposure to serotypes 1a, 1b, 2, 3, 5, 6, 7, 8, 9, 11, 12, 15, 16, 18, 19, 21, or N, but 20% to 30% mortality occurred in vaccinated mice challenge exposed to serotypes 10, 14, 20, or 22. Nonvaccinated control mice died after challenge exposure to all serotypes tested. Vaccinated swine challenge exposed to strain 14B (serotype 9) or strain 2179 (serotype 10) developed localized urticarial lesions at the site of intradermal exposure. Vaccinated swine challenge exposed to serotypes 1a, 1b, 2, 5, 8, 11, 12, 18, 19, or 21 did not have clinical signs of acute erysipelas. Nonvaccinated control swine developed acute generalized erysipelas or localized lesions at the site of intradermal exposure.

Animals↗

Infectivity, disease patterns, and serologic profiles of reovirus serotypes 1, 2, and 3 in infant and weanling mice.

The oronasal median infectious doses of reovirus serotypes 1, 2, and 3 were established in infant and weanling Sencar mice on the basis of disease expression and seroconversion. Infant mice were susceptible to infection with low doses of all three serotypes, whereas weanling mice were comparatively resistant to infection. Uniform transmission of virus to cagemates or mothers of infants did not occur, indicating low contagiousness of all three virus serotypes. The comparative susceptibility of 2-day-old Sencar mice to disease was examined following oronasal inoculation with reovirus 1, 2, or 3. Tissues were collected on days 3, 5, 7, 9, 14, 16, and 21 after inoculation for virus isolation, histologic examination, and serologic analysis. Disease patterns in infant mice were distinctly different among reovirus serotypes. Reovirus 3 induced severe disease, with focal myocarditis, hepatitis, diffuse encephalitis, and generalized lymphoid depletion, whereas reovirus 1 induced a similar pattern, but much milder disease. In contrast, reovirus 2 induced mild transient enteritis without lesions in other organs. Sera from experimentally infected mice were tested in virus serotype-specific enzyme immunoassays. Cross reactivity of antibody among the three virus serotypes was found, but antibody titers were always highest with the homologous antigen. These studies confirm that infant laboratory mice are susceptible to infection with all three serotypes of virus; weanling mice are comparatively resistant to infection and disease; the viruses induce different patterns of disease in infant mice; and infecting virus serotypes can be distinguished serologically by enzyme immunoassay.

Animals↗

Antigenic structure and relationship between serotypes 1 and 2 of Haemophilus paragallinarum.

Antigenic structure and relationship between serotypes 1 and 2 of Haemophilus paragallinarum were analyzed by the rapid plate agglutination and cross-absorption tests. Encapsulated strain forming iridescent colony type of both serotypes 1 and 2 had at least three antigens: heat-labile and trypsin-sensitive (L), heat-labile and trypsin-resistant (HL), and heat-stable and trypsin-resistant (HS). The L was a major antigen located in a surface and divided serologically into three parts: L1, L2, and L3. The L1 was specifici to serotype 1, the L2 was specific to serotype 2, and the L3 was a common antigen shared by serotypes 1 and 2. The HL and HS were common antigens between serotypes. By trypsinization or heating at 121 C, L antigen lost its agglutinability and agglutinin-producing ability. Nonencapsulated organisms forming noniridescent colony type lacked the L antigen. These results suggested that antigenic structure of H paragallinarum serotypes 1 was L1, L3, HL and HS, while serotype 2 was L2, L3, HL, and HS.

Agglutination Tests↗

Nosocomial outbreak of infant rotavirus diarrhea due to the appearance of a new serotype 4 strain.

An outbreak of acute gastroenteritis, involving 30 infants and young children aged 2 months to 4 years, took place in a pediatric ward of the University Hospital of Pavia, Northern Italy, in the period from November 9 to December 1, 1986. Out of the 14 patients examined, ten were found to shed rotavirus with stools. All strains were characterized for serotype, using a monoclonal antibody-based enzyme-linked immunosorbent assay, and for electropherotype, by polyacrylamide gel electrophoresis of genomic RNA. It was shown that a single serotype 4 (subtype 4A) strain spread within the ward from a primary case to seven other patients. The remaining two patients were found to be infected by a serotype 1 strain that was circulating in the same area prior to the outbreak. The clinical symptoms were unusually severe, since significant dehydration was observed in four of the eight serotype 4 rotavirus-infected children. Previous epidemiological studies had shown that since 1983 serotype 4 strains had not been circulating in Pavia, and the electropherotype of the newly circulating serotype 4 strain was different from those observed in 1981-1983. Thus, the severity of the diarrheal disease appeared to be related to the circulation of both a new serotype and a new electropherotype.

Child, Preschool↗

Three serotypes of Norwalk-like virus demonstrated by solid-phase immune electron microscopy.

Solid phase immune electron microscopy (SPIEM) was used to investigate the serological differences between Norwalk-like virus (NLV) strains from five different outbreaks within the United Kingdom. The existence of two previously demonstrated serotypes, Lewis et al. (Journal of Clinical Microbiology 26:938-942, 1988), was confirmed by the use of whole convalescent sera and purified IgM. A third serotype was found to be the agent of two recent hospital outbreaks and could similarly be typed by use of whole sera or pure IgM. Paired sera were available for two of the three serotypes and demonstrated rising antibody levels. These antibody rises were also specific for the infecting serotype. However, two serum pairs from a later outbreak gave antibody rises to all three serotypes, although much higher counts were produced with the infecting serotype. SPIEM is a useful method for distinguishing NLV serotypes and can also be used to detect specific IgM and to demonstrate seroconversion. Cross-reacting antibodies, possibly anamnestic in origin, can occur after natural infection and could cause confusion in typing virus unless further evidence of the identity of the infecting agent is obtained.

Adult↗

Transmission and distribution of African horse sickness virus serotypes in South African zebra.

The prevalences of African horse sickness (AHS) virus serotypes in zebra foals from the Kruger National Park, South Africa were examined for possible associations between serotypes. Serotypes known to cross-react were combined for analysis. The distributions of serotypes between zebra were not always independent; in 7-8 month old zebra positive pairwise associations were observed between 3 serotypes. This could be generated by biological interactions between serotypes or heterogeneity in host-vector transmission. The data were also used to estimate the basic reproduction number, R0. For AHS virus overall, estimates of R0 ranged from 31-68. This underlines the need for a better understanding of serotype transmission and interactions in AHS.

African Horse Sickness↗

Electropherotype heterogeneity within serotypes of human rotavirus strains circulating in Italy. Brief report.

Using solid-phase immune electron microscopy, 126 of 129 human rotavirus (HRV) strains could be serotyped directly in stools collected in Italy during the period 1981-1985. Prevalence was 70.5 per cent for serotype 1, and 13.2 per cent for each of serotypes 2 and 4. No serotype 3 strain was detected. In parallel, for 39 of 61 HRV strains tested the electropherotype of genomic RNA was successfully determined. Different electropherotypes were detected among strains of the same serotype, whereas the same electropherotype was found in HRV strains of different serotypes. Serotyping and electropherotyping of HRV strains appear to be complementary to each other, and both should be used in conjunction for epidemiological surveys.

Child, Preschool↗

Serotypes and mating types of clinical strains of Cryptococcus neoformans isolated in Taiwan.

Twenty-one strains of Cryptococcus neoformans isolated from patients in Taiwan were characterized for serotypes and mating types. Slide agglutination test was performed with 8 factor-specific sera (Iatron Company, Japan) to determine the serotypes. Wheat bran agar (WBA) and malt extract agar (MEA, Wickerham) media were used for the mating tests. Twenty of the isolates were of serotype A, and one was serotype B. Except for 2 strains of serotype A, all of the serotype A strains mated with Filobasidiella neoformans var. neoformans, mating type a. The only serotype B strain mated with F. neoformans var. bacillispora mating type a in MEA medium. These data revealed the low prevalence (1/21; 4.8%) of C. neoformans var. gattii in Taiwan, a subtropically located isoland.

Cryptococcosis↗

Fourteen serotypes of ureaplasma urealyticum (T-mycoplasmas) demonstrated by the complement-dependent mycoplasmacidal test.

Previously, we found that strains of Ureaplasma urealyticum could be divided into 11 serotypes using the mycoplasmacidal (MC) method for serotyping. Subsequent study has shown that two pairs of these strains were sufficiently closely related to cause us to revise the typing scheme into nine distinct serotypes. Other investigators, using different methods to detect serotypic diversity, have found eight serotypes. We have now compared our nine strains with the eight prototype strains developed by Ford and Black, and we have used reciprocal MC titers of antisera for the comparison. Three pairs of strains from our group and the group prepared by Ford and Black were found to be serologically closely related or identical, leaving six serotypes in our series and five in the other series that were serologically unique. Thus 14 serotypes of U. urealyticum have been identified in these studies, and a 15th strain that is apparently serologically distinct from the others, has recently been described in Vancouver.

Serotyping↗

Serotypes of CNF1-producing Escherichia coli strains that cause extraintestinal infections in humans.

The O:K:H serotypes of 137 necrotoxigenic Escherichia coli (NTEC) producing the cytotoxic necrotizing factor type 1 (CNF1) isolated from human extraintestinal infection were determined. Although NTEC producing CNF1 belonged to 58 different serotypes, only 10 of them accounted for 54% of strains. The most common serotypes, in order of frequency, were: O4:K?:H5, O6:K13:H1, O83:K1:H31, O75:K95:H5, O2:K1:H6, O2:K7:H-, O75:K1:H7, O2:K?:H1, O4:K12:H1 and O22:K13:H1. CNF1 strains of serotypes O2:K7:H- and O4:K12:H1 express P-fimbriae, whereas CNF1 strains of serotypes O2:K?:H1, O2:K1:H6 and O75:K95:H5 possess the adhesin responsible for MRHA type III. Among CNF1 strains of serotype O4:K?:H5 there exist some that express P-fimbriae and others that possess MRHA type III. Lastly, the majority of CNF1 strains of serotypes O6:K13:H1, O22:K13:H1, O75:K1:H7 and O83:K1:H31 do not express P-fimbriae nor the adhesin responsible to MRHA type III. Our results show that extraintestinal infections are caused by a limited number of virulent clones, as suggested by the theory of special pathogenicity.

Adhesins, Bacterial↗

Special O:K:H serotypes among enterotoxigenic E. coli strains from diarrhea in adults and children. Occurrence of the CF (colonization factor) antigen and of hemagglutinating abilities.

O:H serotypes previously found to be prevalent among a number of toxigenic strains from several geographic areas were examined for polysaccharide K antigen type. Members of each O:H serotype had the same type of K antigen and were found to be characterized by a certain fermentation pattern. Some O:H serotypes had no K antigen. The serofermentative types defined were: 06:K15:H16, 08:K40:H9, 015:H11, 025:K7:H42, 025:K98:H-, 078:H11, 078:H12, and 0149:H10. Some strains of the last-mentioned serotype, which were suspected of having caused a food-borne infection, had K88. This serotype belongs to the group of strains causing diarrhea in swine. The surface antigen (CF) described as a colonization factor [5] was demonstrated in 078:H-, 078:H11, and 078:H12 strains; but not in any strain of the other serotypes nor in any of 248 strains belonging to 078 but not isolated from cases of human diarrhea. Presence of the CF antigen was correlated with presence of a mannose-resistant ability to cause agglutination of human red cells. Behavior of the other serotypes as regards hemagglutinating abilities was examined and 025:K7:H42 strains were found to be very similar to the 078 strains in this respect.

ABO Blood-Group System↗

Serotypic characterization of group A rotaviruses associated with children's diarrhea in Slovakia.

A total of 368 rotavirus RNA-positive (PAGE) stool samples collected continually during 1992-95 from infants and young children under five years of age hospitalized with acute gastroenteritis were serotyped using an enzyme immunoassay with VP7-specific monoclonal antibodies (ELISA with MAbs) for serotypes G1-G4. The serotype was identified in 106 stool samples (29%). Comparison of electropherotype and serotype profile in individual samples did not show any remarkable correlation. The members of three electropherotypes (A, C, K) belonged to all 4 serotypes. The representatives of two electropherotypes (E, G) and of mixed electropherotype did not react with any of the specific monoclonal antibodies used. The distribution of the serotypes was scored as 52, 13, 14 and 7.5% for G1 G4, respectively, whereas 13% of samples reacted with two or more type-specific monoclonal antibodies. The G1 serotype dominated during the period followed.

Antigens, Viral↗