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[K-serotype analyses of Vibrio parahaemolyticus isolated in northern Taiwan, 1983 through 1993].

From 1983 through 1993, 786 strains of Vibrio parahaemolyticus were collected from food-borne disease outbreaks and sporadic cases of diarrheal illness in northern Taiwan, involving 42 K-serotypes. Five top leading serotypes were K8 (36.8%), K15 (10.8%), K12 (8.7%), K56 (7.9%) and K63 (4.7%). However, a variation of K-serotypes was found during this study period. From 112 food-borne outbreaks associated with this microorganism, only 54 (48.2%) outbreaks were caused by a single serotype, while 58 (51.8%) were caused by multiple K-serotypes. Numbers of outbreaks caused by two, three and more than three K-serotypes were 29 (26%), 16 (14.2%), and 13 (11.6%), respectively. In a special outbreak, eight K-serotypes was found. Outbreaks caused by party caterers were most frequently associated with multiple K-serotypes.

Disease Outbreaks↗

[Extra-intestinal infections caused by non-typhi Salmonella serotypes. 9 yrs' experience].

BACKGROUND: The most characteristic finding in non typhoid Salmonella serotype infection is acute gastroenteritis which is usually benign and self-limiting. However, more serious extraintestinal findings, such as bacteraemia and focal infections localized to any organ may appear. PATIENTS AND METHODS: The objective of this article is to describe the most important characteristics of the extraintestinal infections due to non typhoid Salmonella serotypes observed in our hospital between December 1991 and August 1999. To do so, we reviewed the clinical histories of patients in which extraintestinal non typhoid Salmonella serotypes was isolated, applying a clinicomicrobiological protocol. RESULTS: Salmonella enterica was isolated on 1,543 occasions, of which 25 (1.6%) were Salmonella serotype Typhi and 1,518 (98.3%) non typhoid serotypes. Of the patients with non typhoid serotypes infections, 27 had extraintestinal infection (1.8%). The most frequently isolated was the serotype typhimurium (50% of cases) followed by enteritidis (26% of cases) and hadar (7.1% of cases). Regarding the clinical picture, bacteraemias secondary to gastroenteritis and focal infections occurred with the same frequency (35.7%); the remaining were primary bacteraemias. Eight patients were under 10 years of age and 15 patients were over 60. In 57.1% of the patients there was underlying disease. CONCLUSIONS: Extraintestinal non typhoid Salmonella serotypes infections make up 1.8% of all salmonellosis and most of them occur in childhood or in the elderly. Transitory bacteraemias in the context of acute gastroenteritis occur mainly in children. The greatest risk of acquiring focal infections is associated with the existence of underlying illness or immunodepression.

Adolescent↗

Susceptibility patterns and serotypes of non-typhoidal salmonella in Trinidad.

OBJECTIVE: To study the serotype distribution of non-typhoidal salmonella isolated from clinical specimens and to evaluate their susceptibility patterns to the most commonly prescribed antimicrobial agents used in this region. METHODS: A retrospective study during the period 1993 1999 was carried out at the Eric Williams Medical Sciences Complex Microbiology Laboratory, a teaching hospital in the northwestern part of Trinidad. Strains of non-typhoidal salmonella were serogrouped with polyvalent Salmonella O antiserum and sent to the local Public Health Laboratory for confirmation and further serotyping. RESULTS: A total of 412 strains of non-typhoidal salmonella were serotyped. Of the 13 serotypes, Salmonella enteritidis was the most prevalent 57%, followed by Salmonella typhimurium, 25%, Salmonella virchow 5% and Salmonella heidelberg, 4%. Three hundred and sixty-two (87.5%) non-typhoidal salmonella were isolated from stool, 38 (9%) from blood, 8 (2%) from urine and 4 about 1% from infected wounds. Of the 13 serotypes only Salmonella enteritidis, Salmonella typhimurium, Salmonella virchow and Salmonella heidelberg were simultaneously isolated from blood and stool. One serotype, Salmonella aberdeen, was encountered for the first time in 1993. All isolates were fully sensitive to chloramphenicol, gentamicin, cefuroxime and ceftrioxone. Resistance to ampicillin, augmentin and co-trimoxazole was less than 4%. One third of all non-typhoidal salmonella isolates were resistant to tetracyclines. CONCLUSION: The most prevalent serotypes were Salmonella enteritidis and Salmonella typhimurium which accounted for 82% of all isolates. The prevalence of resistance of non-typhoidal salmonella to antibiotics was very low (5%). Therefore, due to this low prevalence of resistance, we did not provide details of resistance in relation to serotype. However, it was observed that of the 137 non-typhoidal salmonella resistant to tetracycline, more than 80% were Salmonella enteritidis and Salmonella typhimurium strains. Of the 9 antibiotics, tetracycline was the only drug to which one-third of the non-typhoidal salmonella were resistant. Due to the risk of resistance continued surveillance of susceptibility patterns of non-typhoidal salmonella to antibiotics must be maintained and encouraged.

Anti-Bacterial Agents↗

Identification of genes involved in biosynthesis of Actinobacillus pleuropneumoniae serotype 1 O-antigen and biological properties of rough mutants.

Actinobacillus pleuropneumoniae is an important pathogen of swine. Lipopolysaccharide (LPS) has been identified as the major adhesin of A. pleuropneumoniae and it is involved in adherence to porcine respiratory tract cells. We previously generated seven rough LPS mutants of A. pleuropneumoniae serotype 1 by using a mini-Tn10 transposon mutagenesis system [Rioux S, Galarneau C, Harel J et al. Isolation and characterization of mini-Tn10 lipopolysaccharide mutants of Actinobacillus pleuropneumoniae serotype 1. Can J Microbiol 1999; 45: 1017-1026]. The purpose of the present study was to characterize these mutants in order to learn more about LPS O-antigen biosynthesis genes and their organization in A. pleuropneumoniae, and to determine the surface properties and virulence in pigs of these isogenic mutants. By mini-Tn10 insertions in rough mutants, four putative genes (ORF12, ORF16, ORF17, and ORF18) involved in O-antigen biosynthesis in A. pleuropneumoniae serotype 1 were found within a region of 18 ORFs. This region is homologous to the gene cluster of serotype-specific O-polysaccharide biosynthesis from A. actinomycetemcomitans strain Y4 (serotype b). Two mutants showed homology to a protein with identity to glycosyltransferases (ORF12); two others had the mini-Tn10 insertion localized in genes encoding for two distinct proteins with identity to rhamnosyltransferases (ORF16 and ORF17) and three showed homology to a protein which is known to initiate polysaccharide synthesis (ORF18). These four ORFs were also present in A. pleuropneumoniae serotypes 9 and 11 that express an O-antigen that serologically cross-reacts with serotype 1. Evaluation of some biological properties of rough mutants seems to indicate that the absence of O-chains does not appear to have an influence on the virulence of the bacteria in pigs and on the overall surface hydrophobicity, charge and hemoglobin-binding activity, or on LAL activation. An acapsular mutant was included in the present study in order to compare the influence of O-chains and capsule polysaccharides on different cell surface properties. Our data suggest that capsular polysaccharides and not O-chains polysaccharides have a major influence on surface properties of A. pleuropneumoniae serotype 1 and its virulence in pigs.

Actinobacillus pleuropneumoniae↗

Genetic diversity of Streptococcus suis serotypes 2 and 1/2 isolates recovered from carrier pigs in closed herds.

The aim of this study was to compare, by randomly amplified polymorphic DNA (RAPD), the diversity of Streptococcus suis serotypes 1/2 and 2 isolates recovered at slaughter houses from the tonsils of clinically healthy pigs. The pigs belonged to herds with or without clinical signs of S. suis disease. Overall, a low diversity was observed among isolates of serotype 1/2. A representative isolate recovered from a diseased animal presented a relatively high similarity (85%), with most isolates recovered from carrier pigs, from herds either with or without clinical signs of S. suis disease. For serotype 2 isolates, a relatively high degree of heterogeneity was observed in the whole population. Two subpopulations were observed for serotype 2 isolates, which arose from herds with clinical signs. Interestingly, the representative isolate coming from the diseased pig was included in a small closed cluster, with 2 isolates recovered from carrier pigs belonging to the same herd. On the other hand, most of the S. suis serotype 2 isolates originating from herds with no history of S. suis disease, were closely related (90% similarity). Furthermore, they presented different RAPD patterns from those originating from animals from the herd presenting S. suis clinical signs due to this serotype. Results suggest that, in the herds studied, clinical manifestations due to serotype 2 are probably related to the virulence of a specific isolate. Conversely, for the herd affected with serotype 1/2, clinical manifestations of the disease were more likely to be the result of inherent herd factors than the virulence of the specific isolate.

Abattoirs↗

[Antibiotic resistance and serotype distribution of Streptococcus pneumoniae strains isolated from patients at Hacettepe University Medical Faculty].

The emergence of penicillin and multidrug resistant pneumococci has become a global concern. In this study, the in-vitro susceptibilities of 212 Streptococcus pneumoniae strains, isolated from different clinical specimens, to penicillin, erythromycin, cefaclor, cefotaxime, vancomycin, rifampin, tetracycline, chloramphenicol and ciprofloxacine were tested by agar dilution method. Penicillin resistance and intermediate penicillin resistance were found in 7.08% and 41.51% of isolates, respectively. Overall percentages of resistance to erythromycin, cefaclor, cefotaxime, rifampin, tetracycline, chloramphenicol and ciprofloxacine were 13.7%, 17.5%, 2.3%, 2.1%, 18.9%, 2.3% and 1%, respectively. Vancomycin resistance was not detected in the tested strains. All penicillin-susceptible S. pneumoniae isolates were also found to be susceptible to cefaclor, cefotaxime, chloramphenicol and ciprofloxacine. Resistance to other antimicrobial agents was higher among the penicillin-resistant group than the penicillin-susceptible one. Although high level penicillin resistance rate was low, 41.51% of pneumococci were intermediately resistant to penicillin. These results indicated that penicillin resistant S. pneumoniae isolates might probably become a major problem in the near future. Serotyping of penicillin intermediate and resistant S. pneumoniae isolates (n:103) by the capsular swelling test (Quellung reaction) revealed serotypes 19 (n:21), 23 (n:17), 9 (n:8), 14 (n:5), 6 (n:5), 3 (n:4), 15 (n:3), 8 (n:2), 1 (n:2), 20 (n:2) and serotypes 5, 10, 33, 22, 7 and 4 as single isolates. Twenty six S. pneumoniae isolates failed to be serotyped by the antisera set used. All serotype 9 pneumococci strains were found to be multidrug-resistant. Other multidrug-resistant pneumococci strains belonged to serotypes 19, 23 and nontypable group. The serotypes detected among the penicillin intermediate and resistant pneumococci in our study, are among the serotypes covered by 23-valent and 7-valent conjugated pneumococcal vaccine, indicating the possible preventive role of the vaccine against drug-resistant pneumococcal infections.

Anti-Bacterial Agents↗

Serological differentiation of five bluetongue virus serotypes in indirect ELISA.

The serological reactivity in indirect ELISA of five different bluetongue virus (BTV) serotypes (4, 10, 15, 16 & 20) was compared using polyclonal antisera raised against virus particles and an outer structural protein, VP2. Rabbit and sheep antisera against BTV-10 produced higher ELISA values with their homologous antigens than with heterologous serotypes. A hyperimmune rabbit serum specific for virus particles was able to distinguish heterologous serotypes from each other, but a sheep serum from an infected animal was not. An antiserum directed against VP2, the protein responsible for serotype specificity in neutralization tests, was not serotype-specific in ELISA and cross-reacted with other serotypes. The discriminatory ability of a BTV-4 antiserum was improved by cross-absorption with heterologous antigens. This greatly reduced the ELISA signals with heterologous serotypes and produced an antiserum that was effectively serotype-specific.

Bluetongue virus↗

Characterization of monoclonal antibodies against Actinobacillus pleuropneumoniae serotype 5.

Two monoclonal antibodies against Actinobacillus pleuropneumoniae serotype 5, designated as 5MAb-1 and 5MAb-6, were characterized. Enzyme-linked immunosorbent assay-inhibition tests with whole-cell antigens obtained from strains of serotype 1 through 12 of A pleuropneumoniae revealed that 5 MAb-1 bound to only serotype-5 strains. The epitope recognized by 5MAb-1 was a carbohydrate that was sensitive to periodate oxidation and resided on the structure of beta-1,6-linked D-galactose in an O-antigen polysaccharide of serotype-5 lipopolysaccharide. Analysis of these results revealed that the O-antigen polysaccharide of lipopolysaccharide was 1 of the antigenic determinants responsible for the serotype specificity of A pleuropneumoniae. On the other hand, 5MAb-6 reacted with strains of serotype 1 through 10 in varying degrees and its epitope was located on polypeptides sensitive to proteinase K. In an immunoblotting analysis, 5MAb-6 reacted with 2 polypeptide bands, with molecular weights of approximately 41,500 and 28,000, in the outer membrane protein-rich fraction obtained from strains of serotype 1 through 10. These results indicated that outer membrane proteins from several serotype strains of A pleuropneumoniae possessed common antigenic determinants.

Actinobacillus pleuropneumoniae↗

[Prevalence of Candida albican serotypes in blood isolates in Chile, and first report of Candida dubliniensis candidemia].

Our main goal was to determine the prevalence of C. albicans serotypes isolates from blood cultures and identify the presence of C. dubliniensis. We studied 47 strains identified as C. albicans by conventional methods, 28 were isolated from children and 19 from adult patients. The strains were re-identified by standard methods and phenotypic screening as xylose assimilation and growth at 42 degrees C. API ID 32C (bioMérieux) was employed with the C. dubliniensis suspected strains and confirmation was made by molecular fingerprinting using random amplified polymorphic DNA (RAPD). The C. albicans serotype was determined by agglutination with antiserum anti-antigen 6 from cell wall (Candida Check, Iatron Inc., Japan) and the in vitro susceptibilities were evaluated by a microdilution method. From 47 strains, 46 were confirmed as C. albicans, 31 of them (67%) were serotype A. Adult patients presented a high prevalence of serotype A (95%) and children presented a frequency of 52% of the serotype B (p<0.05). We confirmed the identification of C. dubliniensis in one strain isolated from an infant. All serotype B strains were susceptible to fluconazole, itraconazole and amphotericin B. On the other hand, 3% and 6% of serotype A strains were "susceptible dose dependent" to fluconazole and itraconazole, respectively. C. albicans serotype A was predominant in adult candidemia and its distribution was homogenous in children patients. All strains were highly susceptible to antifungals. We report here the first case of C. dubliniensis candidemia in South America.

Adult↗

Evaluation of three panels of monoclonal antibodies for the identification of human rotavirus VP7 serotype by ELISA.

Three panels of monoclonal antibodies used for rotavirus serotype identification by enzyme-linked immunosorbent assay (ELISA) were evaluated at the National Institutes of Health, USA, to identify antibodies suitable for distribution to laboratories involved in WHO-sponsored trials of rotavirus vaccines. Two of the panels were comparably effective in identifying the serotype of each of the human rotavirus reference strains of serotype 1, 2, or 3. In addition, one of the panels included a monoclonal antibody that was effective in identifying strains of serotype 4. However, two different lots of a third, commercially available panel were not effective in identifying the eight strains representing the four serotypes. A third shipment of this panel was therefore tested using revised instructions and, under these conditions, it was effective in serotyping seven of the eight reference strains. It appears that a battery of monoclonal antibodies for each serotype may be required to identify antigenic variants within a serotype. Additional studies are needed to assess the extent of antigenic variation in rotavirus field strains.

Antibodies, Monoclonal↗

Human monoclonal antibodies specific for capsular polysaccharides of Klebsiella recognize clusters of multiple serotypes.

We report the generation and the characterization of a set of human monoclonal antibodies (HmAb) specific for Gram-negative bacteria of Klebsiella pneumoniae. The eight human hybridomas secrete either IgM kappa, IgA1 kappa, or IgA2 kappa antibodies. One HmAb binds bacteria of only one serotype. Five HmAb recognize non-overlapping clusters of 2, 3, or 10 different serotypes. The remaining two HmAb both recognize three serotypes. Two serotypes are recognized by both HmAb, and in addition both HmAb bind one more nonidentical serotype. These results suggest that in man, epitopes are immunodominant, different from serotype-specific determinants detected by conventional rabbit antisera. Screening of clinical isolates revealed that the HmAb recognize not only representative typing strains but also most isolates of the corresponding serotype. In addition, most of the isolates that were non-typable by polyclonal antisera were recognized by one of the HmAb. Fine specificity analyses revealed that all HmAb are highly specific for the isolated capsular polysaccharides (CPS) of bacteria within the corresponding cluster of serotypes. However, the avidity of a HmAb for the different CPS can differ significantly. Taken together, our results suggest that the unequivocal interactions between HmAb and CPS offer the basis for an alternative, better defined classification system, and that passive immunization with a limited number of HmAb may provide a feasible strategy for the protection against the majority of fatal, nosocomial infections with multidrug-resistant strains of K. pneumoniae.

Antibodies, Bacterial↗

[Classify species of Pseudomonas pseudomallei into serotypes].

Species of P. pseudomallei can be classified into two serotypes, serotype I and serotype II, based on whether or not it contains a thermolabile antigen beside a thermostable one. Under the condition of lack of typing serum, by means of serum absorption test, we recognized a strain which contains a major thermolabile antigen. The antigen was purified by Sephadex G-200, and it was used to inoculate rabbits. With the immunoserum at hand, we identified 68 of the domestic chinese strains and 6 of alien strains for serotyping by bi-directional agar diffusion test. The results showed that 68 strains were identified serotype I, 3 strains serotype II, and the remaining 3 comparable with those reported indicating that strains of serotype I were found mostly in Asia, and that the serotype are unrelated to their existing environments, nor that of animal bodies, but connected with their existence in geographic distribution.

China↗

[Streptococcus pneumoniae serotypes in children with chronic inflammatory diseases of the respiratory organs].

During the 7-year period of observation (1982-1988) the serotypes of 276 pneumococcal strains isolated from children with chronic bronchopulmonary diseases were studied. Among the serotypes of pneumococci under study, serotypes 6, 19 and 15 held the leading place and included a half of all typed pneumococci. Dynamic observation on the serotype composition of pneumococci revealed periodic fluctuations in the occurrence of some types/groups. The regional analysis of different serotypes of pneumococci showed the common occurrence of serogroups 6 and 19, as well as some regional features in the circulation of serotypes 6, 10, 3 and rarely occurring serotypes. The study revealed that any new exacerbation of the chronic bronchopulmonary process is caused by pneumococci of some other serotype. Pneumococcal strains, resistant (3.4%) and sensitive (1.8%) to penicillin, were detected; most of them belonged to serogroup 19.

Adolescent↗

[Streptococcus pneumoniae serotypes in children with acute pneumonia and pleurisy].

The study of pneumococci of different serotypes, isolated from patients with acute pneumonia and pleuritis and from healthy children was carried out. Among the pneumococcal serotypes causing pneumonia and pleuritis in children serotypes 1, 6, 19, 14 and 3 were most widely spread and constituted 62.3% of all isolated pneumococci. In young children cases of acute pneumonia and pleuritis were more often induced by serotypes 6 and 14 and in older children, by serotypes 1 and 3. In patients with uncomplicated pneumonia and pleuritis differences in the detected serotypes of pneumococci were observed, and the disease course differed in severity. Serotypes 14, 3 and 3 induced destructive processes in the lungs more often than other serotypes. Monitoring of the sensitivity of pneumococci to antibiotics showed that most of the strains retained high sensitivity to penicillin and ampicillin. In most cases the detected resistant pneumococcal strains belonged to serogroup 19.

Acute Disease↗

Campylobacter jejuni/coli in commercially reared beagles: prevalence and serotypes.

A 5 year longitudinal study involving 187 commercially reared beagles from three suppliers was undertaken to determine prevalence and serotypes of Campylobacter jejuni and C. coli. Campylobacter jejuni or C. coli was isolated from the feces in 62 of 177 asymptomatic beagles and 8 of 10 dogs with diarrhea for an overall prevalence of 37%. A total of 36 isolates were serotyped on the basis of thermostable antigens with 20 antisera prepared against frequently occurring serotypes isolated from humans with campylobacter associated enteritis (15 C. jejuni, 5 C. coli serotypes). Of these isolates, 17 (47%) serotyped with antisera to 7 C. jejuni serotypes frequently isolated in human cases of enteric campylobacteriosis (serotypes 1, 4, 10, 16, 18, 19, 37). One C. coli reacted to antisera 24, 34, 37, one strain of C. coli to antisera type 37, and another C. coli to antisera type 34. All three C. coli belonged to serotypes frequently encountered in diarrheic human patients.

Animals↗

Neutralization microtest with human coxsackievirus and echovirus serotypes.

Sets of 600 single sera from healthy individuals of various age and 458 paired sera from patients treated for diseases of various etiology were examined using a neutralization microtest technique employing prototype collection strains CA9, CB1-CB5, E1-E9, E11-E14, E17, E19, E24 and E26. Totally, 34,862 monotype neutralization tests were carried out in this study. In the set of single serum samples the lowest proportion of sera reacting with any of the enterovirus serotypes used was encountered in the group of youngest children up to the age of 2 years. This group of children showed also the highest proportion of sera free of type-specific antibody. Taken together, these sera reacted most frequently with serotypes CA9 and CB4. In the set of paired sera significant rises in antibody titre to one enterovirus serotype were recorded in 105 instances, in association with simultaneous nonsignificant rises against one or several other serotypes in 64 instances. In additional 12 paired sera there was evidenced a significant rise of antibody titre to more than one serotypes, combined in 8 of these with simultaneous nonsignificant rises to further serotypes. This neutralization microtest technique with a set of enterovirus serotypes is believed to represent a servicable diagnostic tool in determining the cause of enteroviral infection, in spite of the estimated 10% failure to establish the exact serotype responsible. The results yielded by the serologic examination of single serum samples are not considered as reflecting the actual seroconversion rates in the general population.

Adolescent↗

Serological survey of infectious bursal disease virus serotypes 1 and 2 in California turkeys.

The prevalence of two infectious bursal disease (IBD) viruses--serotype 1, of chicken origin, and serotype 2, of turkey origin--was studied in California turkeys. Serum samples were collected from 15 turkey flocks representing nine counties. The virus-neutralization test was used. Overall, only 15% of 342 samples were positive for serotype 1, whereas 89% were positive for serotype 2. However, 12 out of 15 flocks had a least one sample positive for serotype 1, and 14 out of 15 flocks had at least one sample positive for serotype 2. Flocks with antibodies to serotype 1 had low geometric mean titers (GMTs) (1.5 to 8.8) and low-to-medium prevalence rates (5 to 53.3%). Flocks with antibodies to serotype 2 had high GMTs (36.8 to 2048) and high prevalence rates (60 to 100%). Results of this study lead us to question the efficacy of IBD vaccination in turkeys.

Animals↗

A comparison of an australian bluetongue virus isolate (CSIRO 19) with other bluetongue virus serotypes by cross-hybridization and cross-immune precipitation.

No major differences in size were observed when both the double-stranded RNA and the polypeptides of the Australian bluetongue virus (BTV) isolate CSIRO 19 (BTV-20) were compared with those of other BTV serotypes such as BTV-10 and BTV-4. Minor capsid polypeptide P6 of both BTV-20 and BTV-4, which electrophoreses as a single band on continuous phosphate buffered gels, in separated into 2 distinct bands on discontinuous glycine-buffered gels. This was not the case with BTV-10. Cross-immune precipitation of BTV-20 with BTV-10, BTV-17, BTV-4 and BTV-3 indicated strong immunological cross-reaction of the group-specific antigen P7 of the different serotypes. There was also some cross-immune precipitation of the serotype-specific polypeptide P2 of BTV-20 and BTV-4. This result is in agreement with the observed cross neutralization of these 2 viruses. The main distinction between BTV-20 and the other BTV serotypes was observed in cross-hybridization experiments. The homology between the nucleic acid of BTV-20 and other BTV serotypes was less than 30%, whereas homology normally found between BTV serotypes is at least 70%. The hybridization products of the different BTV serotypes were analysed by electrophoresis and fluorography. Two main hybrid segments were observed in all heterologous hybridizations with BTV-20 as a compared with 7 hybrid segments in hybridizations between BTV-4 and BTV-10. In order to determine from which genome segment of BTV-20 these 2 hybrid segments were derived, the hybridizations were carried out with individually purified double-stranded RNA segments. These results indicate that the 2 segments of BTV-20 that show the largest homology to corresponding segments of a heterologous BTV serotype are No. 7 and 10.

Bluetongue virus↗